title: PeerJ description: Articles published in PeerJ link: https://peerj.com/articles/index.rss3?journal=peerj&month=2023-03 creator: info@peerj.com PeerJ errorsTo: info@peerj.com PeerJ language: en title: Machine learning aided multiscale modelling of the HIV-1 infection in the presence of NRTI therapy link: https://peerj.com/articles/15033 last-modified: 2023-03-31 description: Human Immunodeficiency Virus (HIV) is one of the most common chronic infectious diseases in humans. Extending the expected lifetime of patients depends on the use of optimal antiretroviral therapies. Emergence of the drug-resistant strains can reduce the effectiveness of treatments and lead to Acquired Immunodeficiency Syndrome (AIDS), even with antiretroviral therapy. Investigating the genotype-phenotype relationship is a crucial process for optimizing the therapy protocols of the patients. Here, a mathematical modelling framework is proposed to address the impact of existing mutations, timing of initiation, and adherence levels of nucleotide reverse transcriptase inhibitors (NRTIs) on the evolutionary dynamics of the virus strains. For the first time, the existing Stanford HIV drug resistance data have been combined with a multi-strain within-host ordinary differential equation (ODE) model to track the dynamics of the most common NRTI-resistant strains. Overall, the D4T-3TC, D4T-AZT and TDF-D4T drug combinations have been shown to provide higher success rates in preventing treatment failure and further drug resistance. The results are in line with the genotype-phenotype data and pharmacokinetic parameters of the NRTI inhibitors. Moreover, we show that the undetectable mutant strains at the diagnosis have a significant effect on the success/failure rates of the NRTI treatments. Predictions on undetectable strains through our multi-strain within-host model yielded the possible role of viral evolution on the treatment outcomes. It has been recognized that the improvement of multi-scale models can contribute to the understanding of the evolutionary dynamics, and treatment options, and potentially increase the reliability of genotype-phenotype models. creator: Huseyin Tunc creator: Murat Sari creator: Seyfullah Kotil uri: https://doi.org/10.7717/peerj.15033 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Tunc et al. title: Prognostic value of apparent diffusion coefficient in neuroendocrine carcinomas of the uterine cervix link: https://peerj.com/articles/15084 last-modified: 2023-03-31 description: ObjectivesThis research was designed to examine the associations between the apparent diffusion coefficient (ADC) values and clinicopathological parameters, and to explore the prognostic value of ADC values in predicting the International Federation of Gynecology and Obstetrics (FIGO) stage and outcome of patients suffering from neuroendocrine carcinomas of the uterine cervix (NECCs).MethodsThis retrospective study included 83 patients with NECCs, who had undergone pre-treatment magnetic resonance imaging (MRI) between November 2002 and June 2019. The median follow-up period was 50.7 months. Regions of interest (ROIs) were drawn manually by two radiologists. ADC values in the lesions were calculated using the Functool software. These values were compared between different clinicopathological parameters groups. The Kaplan–Meier approach was adopted to forecast survival rates. Prognostic factors were decided by the Cox regression method.ResultsIn the cohort of 83 patients, nine, 42, 23, and nine patients were in stage I, II, III, and IV, respectively. ADCmean, ADCmax, and ADCmin were greatly lower in stage IIB–IVB than in stage I–IIA tumours, as well as in tumours measuring ≥ 4 cm than in those < 4 cm. ADCmean, FIGO stage, and age at dianosis were independent prognostic variables for the 5-year overall survival (OS). ADCmin, FIGO stage, age at diagnosis and para-aortic lymph node metastasis were independent prognostic variables for the 5-year progression-free survival (PFS) in multivariate analysis. For surgically treated patients (n = 45), ADCmax was an independent prognostic parameter for both 5-year OS and 5-year PFS.ConclusionsADCmean, ADCmin, and ADCmax are independent prognostic factors for NECCs. ADC analysis could be useful in predicting the survival outcomes in patients with NECCs. creator: Jian Chen creator: Ning Ma creator: Mingyao Sun creator: Li Chen creator: Qimin Yao creator: XingFa Chen creator: Cuibo Lin creator: Yongwei Lu creator: Yingtao Lin creator: Liang Lin creator: Xuexiong Fan creator: Yiyu Chen creator: Jingjing Wu creator: Haixin He uri: https://doi.org/10.7717/peerj.15084 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Chen et al. title: Leukocyte telomere length and bipolar disorder risk: evidence from Mendelian randomization analysis link: https://peerj.com/articles/15129 last-modified: 2023-03-31 description: ObjectiveWe aim to test whether leukocyte telomere length (LTL) is causally associated with the risk of bipolar disorder (BD) using the Mendelian randomization (MR) method.MethodsResults of a genome-wide association study (GWAS) conducted with 472,174 individuals of European descent were used to screen for single-nucleotide polymorphisms (SNPs) related with LTL traits. Summary-level data for BD (7,647 cases and 27,303 controls) were obtained from UK Biobank. An inverse-variance-weighted (IVW) method was employed as the primary MR analysis. Sensitivity analyses were conducted via MR-Egger, maximum likelihood, MR-pleiotropy residual sum outlier (MR-PRESSO), and MR-robust adjusted profile score (MR-RAPS) methods. Finally, the MR Steiger test was utilized to validate the hypothesized relationship between exposure and outcome.ResultsTwo-sample MR analysis revealed inverse relationships between genetically predicted LTL and BD risk (IVW OR [odds ratio] = 0.800, 95% CI [0.647–0.989] P = 0.039). Genetically predicted LTL exhibits a consistent connection with BD across five MR methods. Sensitivity analyses showed that the genetically determined effect of LTL on BD was stable and reliable. Furthermore, the MR Steiger test demonstrated that LTL was causal for BD rather than the opposite (P < 0.001).ConclusionOur findings show that genetically determined LTL reduces the risk of BD. More research is required to clarify the mechanisms underlying this apparent causal connection. In addition, these findings may be useful for developing strategies for the prevention and treatment of BD. creator: Likui Lu creator: Hongtao Zeng creator: Bangbei Wan creator: Miao Sun uri: https://doi.org/10.7717/peerj.15129 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Lu et al. title: Predicting the geographic distribution of ancient Amazonian archaeological sites with machine learning link: https://peerj.com/articles/15137 last-modified: 2023-03-31 description: Amazonia has as least two major centers of ancient human social complexity, but the full geographic extents of these centers remain uncertain. Across the southern rim of Amazonia, over 1,000 earthwork sites comprised of fortified settlements, mound villages, and ditched enclosures with geometric designs known as geoglyphs have been discovered. Qualitatively distinct and densely located along the lower stretches of major river systems and the Atlantic coast are Amazonian Dark Earth sites (ADEs) with deep anthropogenic soils enriched by long-term human habitation. Models predicting the geographic extents of earthworks and ADEs can assist in their discovery and preservation and help answer questions about the full degree of indigenous landscape modifications across Amazonia. We classify earthworks versus ADEs versus other non-earthwork/non-ADE archaeological sites with multi-class machine learning algorithms using soils, climate, and distances to rivers of different types and sizes as geospatial predictors. Model testing is done with spatial cross-validation, and the best model at the optimal spatial scale of 1 km has an Area Under the Curve of 0.91. Our predictive model has led to the discovery of 13 new geoglyphs, and it pinpoints specific areas with high probabilities of undiscovered archaeological sites that are currently hidden by rainforests. The limited, albeit impressive, predicted extents of earthworks and ADEs means that other non-ADE/non-earthwork sites are expected to predominate most of Western and Northern Amazonia. creator: Robert S. Walker creator: Jeffrey R. Ferguson creator: Angelica Olmeda creator: Marcus J. Hamilton creator: Jim Elghammer creator: Briggs Buchanan uri: https://doi.org/10.7717/peerj.15137 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Walker et al. title: Assessing arthropod diversity metrics derived from stream environmental DNA: spatiotemporal variation and paired comparisons with manual sampling link: https://peerj.com/articles/15163 last-modified: 2023-03-31 description: BackgroundBenthic invertebrate (BI) surveys have been widely used to characterize freshwater environmental quality but can be challenging to implement at desired spatial scales and frequency. Environmental DNA (eDNA) allows an alternative BI survey approach, one that can potentially be implemented more rapidly and cheaply than traditional methods.MethodsWe evaluated eDNA analogs of BI metrics in the Potomac River watershed of the eastern United States. We first compared arthropod diversity detected with primers targeting mitochondrial 16S (mt16S) and cytochrome c oxidase 1 (cox1 or COI) loci to that detected by manual surveys conducted in parallel. We then evaluated spatial and temporal variation in arthropod diversity metrics with repeated sampling in three focal parks. We also investigated technical factors such as filter type used to capture eDNA and PCR inhibition treatment.ResultsOur results indicate that genus-level assessment of eDNA compositions is achievable at both loci with modest technical noise, although database gaps remain substantial at mt16S for regional taxa. While the specific taxa identified by eDNA did not strongly overlap with paired manual surveys, some metrics derived from eDNA compositions were rank-correlated with previously derived biological indices of environmental quality. Repeated sampling revealed statistical differences between high- and low-quality sites based on taxonomic diversity, functional diversity, and tolerance scores weighted by taxon proportions in transformed counts. We conclude that eDNA compositions are efficient and informative of stream condition. Further development and validation of scoring schemes analogous to commonly used biological indices should allow increased application of the approach to management needs. creator: Aaron A. Aunins creator: Sara J. Mueller creator: Jennifer A. Fike creator: Robert S. Cornman uri: https://doi.org/10.7717/peerj.15163 license: https://creativecommons.org/publicdomain/zero/1.0/ rights: © 2023 Aunins et al. title: Comparison of body pressure distribution in healthy subjects between bubble wrap and an emergency mattress laid on a cardboard bed: a randomized controlled crossover trial link: https://peerj.com/articles/15173 last-modified: 2023-03-31 description: BackgroundIt has been pointed out that the poor environment of evacuation shelters causes health problems and disaster-related deaths among evacuees, and we are concerned that their environment will deteriorate, particularly during a large-scale disaster due to a shortage of daily necessities. In Japan, evacuees usually slept on floors with futons until the Great East Japan Earthquake, but cardboard beds were installed in evacuation shelters. Previous studies have suggested that cardboard beds can reduce cold air transmission from the floor. We have reported that a cardboard bed can have a low-contact pressure dispersion capacity and cannot reduce musculoskeletal strain, unlike a futon or mattress. In the Great East Japan Earthquake, 33% of disaster-related deaths were reported to have been caused by physical or mental fatigue due to living in evacuation shelters. When a large-scale disaster such as the Nankai Trough Earthquake generates huge numbers of evacuees, the supply of mattresses for evacuees will be very difficult. Therefore, we considered potential alternatives that could be produced in large quantities over a short period. Bubble wrap, with very lightweight and waterproofing, could be a good candidate for mattress replacement. This study aimed to investigate the improvement in body pressure distribution and pressure-sensing area when using bubble wrap.MethodsTwenty-seven healthy subjects allocated to sequences A and B with different intervention order were laid in supine and lateral positions on a cardboard bed without a mattress, bubble wrap, or air mattress: the mattress-body contact pressure and contour areas were measured, and subjective firmness and comfort during these conditions were also investigated using the visual analog scale (VAS). Acquired data were analyzed using a linear mixed-effects model and Bonferroni’s post-hoc test, and P < 0.05 was considered statistically significant.ResultsThe mattress-body contact pressure and contour area showed significant differences with and without air mattresses. With the air mattresses, the pressure in the supine position decreased by 34%, and that in the lateral position decreased by 13%. However, the four-fold bubble wrap did not improve the mattress-body contact pressure and contour area; the change ratios were within 5% compared to the cardboard bed. However, there were significant differences in subjective firmness and comfort using the VAS among all experimental positions.ConclusionOur study showed that bubble wrap could not significantly improve body pressure concentration and may not be a satisfactory substitute for air mattresses. Because of the improvement in subjective firmness and comfort with the bubble wrap, using it for an extended period may affect the incidence of back pain in evacuees. Finally, we hypothesize that the body pressure dispersion of the bubble wrap may be improved by changing the air-filling rate and the size of the air bubbles. creator: Seiji Hamanishi creator: Yukiko Asada creator: Yu Ikushima creator: Yurika Ikeda creator: Mai Chinushi uri: https://doi.org/10.7717/peerj.15173 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Hamanishi et al. title: miR-126 promotes M1 to M2 macrophage phenotype switching via VEGFA and KLF4 link: https://peerj.com/articles/15180 last-modified: 2023-03-31 description: BackgroundMacrophage polarization and microRNA play crucial roles in the development of atherosclerosis (AS). The M1 macrophage phenotype contributes to the formation of plaques, while the M2 macrophage phenotype resolves inflammation and promotes tissue repair. MiR-126 has been found to play a role in regulating macrophage polarization in the context of AS. However, the exact mechanism of miR-126 requires further research.MethodsThe foam cell model was established by stimulating THP-1 with oxidized low-density lipoprotein (ox-LDL). We transfected foam cells with miR-126 mimic and its negative control. The transfection of miR-126 was implemented by riboFECT CP transfection kit. The levels of miR-126 and M1/M2 associated genes in foam cells were quantified using reverse transcription-quantitative PCR (RT-qPCR). Additionally, the expressions of CD86+ and CD206+ cells in foam cells were determined by flow cytometry. Western blotting and RT-qPCR were used to determine the protein and mRNA levels of the vascular endothelial growth factor A (VEGFA) and the transcriptional regulator Krüppel-like factor 4 (KLF4), respectively. Additionally, we detected endothelial cell migration after co-culturing endothelial cells and macrophages. MG-132 was used to indirectly activate the expression of VEGFA, and the expression of KLF4 was also evaluated.ResultsThe activation of apoptosis and production of foam cells were boosted by the addition of ox-LDL. We transfected foam cells with miR-126 mimic and its negative control and observed that miR-126 greatly suppressed foam cell development and inhibited phagocytosis. Moreover, it caused pro-inflammatory M1 macrophages to switch to the anti-inflammatory M2 phenotype. This was reflected by the increase in anti-inflammatory gene expression and the decrease in pro-inflammatory gene expression. Additionally, miR-126 dramatically decreased the expressions of VEGFA and KLF4. The protein-protein interaction network analysis showed a significantly high correlation between miR-126, VEGFA, and KLF4. MiR-126 may also promote EC migration by activating macrophage PPAR γ expression and effectively suppressing macrophage inflammation. MG-132 indirectly activated the expression of VEGFA, and the expression of KLF4 also significantly increased, which indicates a direct or indirect relationship between VEGFA and KLF4.ConclusionOur study shows that miR-126 can reverse ox-LDL-mediated phagocytosis and apoptosis in macrophages. Consequently, the potential role of miR-126 was manifested in regulating macrophage function and promoting vascular endothelial migration. creator: Xinyang Shou creator: Yimin Wang creator: Qingyu Jiang creator: Jun Chen creator: Qiang Liu uri: https://doi.org/10.7717/peerj.15180 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Shou et al. title: Soil water depletion patterns in rainfed apple orchards and wheat fields link: https://peerj.com/articles/15098 last-modified: 2023-03-30 description: Agricultural production in the Weibei rainfed highland, Northwest China, is challenged by severe drought and water shortages. While the land use pattern has shifted gradually from crop production to orchard farming in Weibei, little is known about the influence of fruit industry development on regional water resources and the rationality of planting orchards. Here, we characterized soil water depletion patterns in rainfed orchards and farmlands to evaluate the occurrence of soil desiccation under land use conversion from farmlands to orchards in Weibei. Soil moisture dynamics were monitored in the 0–150 cm soil profiles of different aged Red Fuji apple orchards (young: 7 years, mature: 13 years, old: 22 years) and long-term cultivated winter wheat fields. We measured soil moisture content by oven-drying method in the middle of each month during the growing season of apple trees (March–September 2019). The over-depletion and depletion of soil water were analyzed to evaluate water stress and differential water depletion by distinct vegetation, respectively. The soil desiccation index was used to determine the occurrence of dry soil layers. Water stress was only observed at the 0–70-cm soil depths in the old orchards (mid-June) and farmlands (mid-May–mid-July). Water depletion took place at deeper depths for longer periods in the older orchards than in the younger orchards. Soil desiccation was absent in the young orchards, with mild desiccation at the 0–80-cm soil depths in the mature and old orchards in mid-June. The desiccation intensity was mild at the 0–60-cm soil depths in mid-April–mid-May, intense at the 0–150-cm soil depths in mid-June, and moderate at the 20–150-cm soil depths in mid-July. Results of this study demonstrate the mitigation of water stress and soil desiccation following conversion from wheat fields to apple orchards, which verifies the rationality of planting orchards in the rainfed highland area. Our findings provide strong support for developing a novel model of agro-industrial development, ecological construction, and sustainable economy in the vast arid and semi-arid areas of Northwest China. creator: Lu Zhang creator: Yiquan Wang creator: Zenghui Sun uri: https://doi.org/10.7717/peerj.15098 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Zhang et al. title: The prevalence of clarithromycin-resistant Helicobacter pylori isolates: a systematic review and meta-analysis link: https://peerj.com/articles/15121 last-modified: 2023-03-30 description: BackgroundKnowledge of global clarithromycin (CLA)-resistant rates of Helicobacter pylori (H. pylori) is crucial for decision of the most appropriate eradication therapies with good clinical outcomes. Therefore, this review and meta-analysis aimed to evaluate the global prevalence of the CLA resistance in H. pylori to provide some guidance for selecting the first-line antibiotics.MethodA comprehensive search was performed for relevant literature until April 2021 in PubMed, Embase, and Web of Science databases. Freeman-Tukey double arcsine transformation was performed to estimate the weighted pooled prevalence of resistance.ResultsThe meta-analysis included 248 articles. The prevalence of CLA-resistant H. pylori was 27.53% (95% CI [25.41–29.69]). The heterogeneity between reports was significant (I2 = 97.80%, P < 0.01). The resistance rate increased from 24.28% in 2010–2017 to 32.14% in 2018–2021 (P < 0.01). Iran, with 38 articles, has the most report. Nevertheless, Switzerland, Portugal, and Israel had the highest resistance rates (67.16%, 48.11%, and 46.12%, respectively). The heterogeneity between the continents and the antimicrobial susceptibility methods also interpreted standard guidelines and breakpoints was insignificant (P > 0.05).ConclusionOverall CLA resistance rate was 27.53%, worldwide. The difference in CLA resistance rate among the included studies can be due to several reasons such as differences in antibiotic prescription rates in various geographic areas, use of different breakpoints or inaccurate criteria in performed studies, and the emergence of multidrug-resistant (MDR) strains. creator: Mohammad Sholeh creator: Saeed Khoshnood creator: Taher Azimi creator: Jasem Mohamadi creator: Vahab Hassan Kaviar creator: Marzieh Hashemian creator: Somayeh Karamollahi creator: Nourkhoda Sadeghifard creator: Hedayat Heidarizadeh creator: Mohsen Heidary creator: Morteza Saki uri: https://doi.org/10.7717/peerj.15121 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Sholeh et al. title: Transcriptome of the pygmy grasshopper Formosatettix qinlingensis (Orthoptera: Tetrigidae) link: https://peerj.com/articles/15123 last-modified: 2023-03-30 description: Formosatettix qinlingensis (Zheng, 1982) is a tiny grasshopper endemic to Qinling in China. For further study of its transcriptomic features, we obtained RNA-Seq data by Illumina HiSeq X Ten sequencing platform. Firstly, transcriptomic analysis showed that transcriptome read numbers of two female and one male samples were 25,043,314, 24,429,905, and 25,034,457, respectively. We assembled 65,977 unigenes, their average length was 1,072.09 bp, and the length of N50 was 2,031 bp. The average lengths of F. qinlingensis female and male unigenes were 911.30 bp, and 941.82 bp, and the N50 lengths were 1,745 bp and 1,735 bp, respectively. Eight databases were used to annotate the functions of unigenes, and 23,268 functional unigenes were obtained. Besides, we also studied the body color, immunity and insecticide resistance of F. qinlingensis. Thirty-nine pigment-related genes were annotated. Some immunity genes and signaling pathways were found, such as JAK-STAT and Toll-LIKE receptor signaling pathways. There are also some insecticide resistance genes and signal pathways, like nAChR, GST and DDT. Further, some of these genes were differentially expressed in female and male samples, including pigment, immunity and insecticide resistance. The transcriptomic study of F. qinlingensis will provide data reference for gene prediction and molecular expression study of other Tetrigidae species in the future. Differential genetic screening of males and females provides a basis for studying sex and immune balance in insects. creator: Yuxin Liu creator: Xuejuan Li creator: Liliang Lin uri: https://doi.org/10.7717/peerj.15123 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Liu et al. title: Changes in soil carbon, nitrogen, and phosphorus in Pinus massoniana forest along altitudinal gradients of subtropical karst mountains link: https://peerj.com/articles/15198 last-modified: 2023-03-30 description: Changes in altitude have a long-term and profound impact on mountain forest ecosystems. However, there have been few reports on changes in soil carbon, nitrogen, and phosphorus contents (SCNPC) along altitudinal gradients in subtropical karst mountain forests, as well as on the factors influencing such changes. We selected five Pinus massoniana forests with an altitudinal gradient in the karst mountain area of Southwest China as research objects and analyzed the changes in SCNPC along the altitudinal gradient, as well as the influencing factors behind these changes. Soil organic carbon, total nitrogen, and available nitrogen contents first increased and then decreased with increasing altitude, whereas the contents of total phosphorus and available phosphorus showed no obvious trend. In the karst mountain P. massoniana forest, SCNPC in the topsoil is most significantly affected by total glomalin-related soil protein (TG) and soil moisture content (SMC) (cumulative explanatory rate was 45.28–77.33%), indicating that TG and SMC are important factors that affect SCNPC in the karst mountain P. massoniana forest. In addition, the main environmental factors that affect SCNPC in the subsoil showed significant differences. These results may provide a better scientific reference for the sustainable management of the subtropical mountain P. massoniana forest. creator: Kun Nie creator: Ming Xu creator: Jian Zhang uri: https://doi.org/10.7717/peerj.15198 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Nie et al. title: Gram-negative ESKAPE bacteria bloodstream infections in patients during the COVID-19 pandemic link: https://peerj.com/articles/15007 last-modified: 2023-03-29 description: Bloodstream infections due to bacteria are a highly consequential nosocomial occurrences and the organisms responsible for them are usually multidrug-resistant. The aims of this study were to describe the incidence of bacteremia caused by Gram-negative ESKAPE bacilli during the COVID-19 pandemic and characterize the clinical and microbiological findings including antimicrobial resistance. A total of 115 Gram-negative ESKAPE isolates were collected from patients with nosocomial bacteremia (18% of the total bacteremias) in a tertiary care center in Mexico City from February 2020 to January 2021. These isolates were more frequently derived from the Respiratory Diseases Ward (27), followed by the Neurosurgery (12), Intensive Care Unit (11), Internal Medicine (11), and Infectious Diseases Unit (7). The most frequently isolated bacteria were Acinetobacter baumannii (34%), followed by Klebsiella pneumoniae (28%), Pseudomonas aeruginosa (23%) and Enterobacter spp (16%). A. baumannii showed the highest levels of multidrug-resistance (100%), followed by K. pneumoniae (87%), Enterobacter spp (34%) and P. aeruginosa (20%). The blaCTX-M-15 and blaTEM-1 genes were identified in all beta-lactam-resistant K. pneumoniae (27), while blaTEM-1 was found in 84.6% (33/39) of A. baumannii isolates. The carbapenemase gene blaOXA-398 was predominant among carbapenem-resistant A. baumannii (74%, 29/39) and blaOXA-24was detected in four isolates. One P. aeruginosa isolate was blaVIM-2 gene carrier, while two K. pneumoniae and one Enterobacter spp were blaNDM gene carriers. Among colistin-resistant isolates mcr-1 gene was not detected. Clonal diversity was observed in K. pneumoniae, P. aeruginosa and Enterobacter spp. Two outbreaks caused by A. baumannii ST208 and ST369 were detected, both belonging to the clonal complex CC92 and IC2. A. baumannii was associated with a death rate of 72% (28/32), most of them (86%, 24/28) extensively drug-resistant or pandrug-resistant isolates, mainly in patients with COVID-19 (86%, 24/28) in the Respiratory Diseases Ward. A. baumannii isolates had a higher mortality rate (72%), which was higher in patients with COVID-19. There was no statistically significant association between the multidrug-resistant profile in Gram-negative ESKAPE bacilli and COVID-19 disease. The results point to the important role of multidrug-resistant Gram-negative ESKAPE bacteria causing bacteremia in nosocomial settings before and during the COVID-19 epidemic. Additionally, we were unable to identify a local impact of the COVID-19 pandemic on antimicrobial resistance rates, at least in the short term. creator: María Dolores Alcántar-Curiel creator: Manuel Huerta-Cedeño creator: Ma Dolores Jarillo-Quijada creator: Catalina Gayosso-Vázquez creator: José Luis Fernández-Vázquez creator: María Luisa Hernández-Medel creator: Manuelita Zavala-Pineda creator: Miguel Ángel Morales-Gil creator: Verónica Alejandra Hernández-Guzmán creator: Manuel Ismael Bolaños-Hernández creator: Silvia Giono-Cerezo creator: José Ignacio Santos-Preciado uri: https://doi.org/10.7717/peerj.15007 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Alcántar-Curiel et al. title: Evaluation of a dill (Anethum graveolens L.) gene bank germplasm collection using multivariate analysis of morphological traits, molecular genotyping and chemical composition to identify novel genotypes for plant breeding link: https://peerj.com/articles/15043 last-modified: 2023-03-29 description: Dill (Anethum graveolens L.) is an aromatic herb widely used in the food industry, with several commercial cultivars available with different qualitative characteristics. Commercial cultivars are usually preferred over landraces due to their higher yield and also the lack of improved landraces than can be commercialized. In Greece, however, traditional dill landraces are cultivated by local communities. Many are conserved in the Greek Gene Bank and the aim here was to investigate and compare the morphological, genetic, and chemical biodiversity of twenty-two Greek landraces and nine modern/commercial cultivars. Multivariate analysis of the morphological descriptors, molecular markers, and essential oil and polyphenol composition revealed that the Greek landraces were clearly distinguished compared with modern cultivars at the level of phenological, molecular and chemical traits. Landraces were typically taller, with larger umbels, denser foliage, and larger leaves. Plant height, density of foliage, density of feathering as well as aroma characteristics were desirable traits observed for some landraces, such as T538/06 and GRC-1348/04, which were similar or superior to those of some commercial cultivars. Polymorphic loci for inter-simple sequence repeat (ISSR) and start codon targeted (SCoT) molecular markers were 76.47% and 72.41% for landraces, and 68.24% and 43.10% for the modern cultivars, respectively. Genetic divergence was shown, but not complete isolation, indicating that some gene flow may have occurred between landraces and cultivars. The major constituent in all dill leaf essential oils was α-phellandrene (54.42–70.25%). Landraces had a higher α-phellandrene and dill ether content than cultivars. Two dill landraces were rich in chlorogenic acid, the main polyphenolic compound determined. The study highlighted for the first-time Greek landraces with desirable characteristics regarding quality, yield, and harvest time suitable for breeding programs to develop new dill cultivars with superior features. creator: Kalliopi Kadoglidou creator: Catherine Cook creator: Anastasia Boutsika creator: Eirini Sarrou creator: Ifigeneia Mellidou creator: Christina Aidonidou creator: Ioannis Grigoriadis creator: Andrea Angeli creator: Stefan Martens creator: Vasiliki Georgiadou creator: Theodoros Moysiadis creator: Parthenopi Ralli creator: Ioannis Mylonas creator: Nikolaos Tourvas creator: Michail Michailidis creator: Apostolos Kalivas creator: Eleni Maloupa creator: Ioannis Ganopoulos creator: Aliki Xanthopoulou uri: https://doi.org/10.7717/peerj.15043 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Kadoglidou et al. title: Atypical ‘long-tailed’ cockroaches arose during Cretaceous in response to angiosperm terrestrial revolution link: https://peerj.com/articles/15067 last-modified: 2023-03-29 description: Typical cockroaches are flat, broad, with large pronotum and wings covering the body. This conserved morphotype dates back to the Carboniferous, during which the ancestral cockroaches, or roachoids, originated. On the other hand, the ovipositor of cockroaches gradually reduced during the Mesozoic, coupled with a major shift of reproductive strategy. By the Cretaceous, long external ovipositors became rare, most cockroaches used very short or even hidden internal ovipositors to fabricate egg cases (oothecae), which is an innovation for egg protection. Here, we describe two cockroaches from mid-Cretaceous Myanmar amber: Ensiferoblatta oecanthoides gen. et sp. nov. (Ensiferoblattidae fam. nov.) and Proceroblatta colossea gen. et sp. nov. They are slim, elongate, fusiform, with longitudinal pronotum, and have long external ovipositors. The combination of these traits represents a unique morphotype, which resembles crickets and katydids (Ensifera) more than general cockroaches. Ensiferoblatta and Proceroblatta may be arboreal, feeding on and/or laying eggs into certain angiosperms that newly emerged. Their open habit causes latent impairment to viability, and may contribute to their extinction. These new taxa are the youngest members of the ancient, extinct group of cockroaches, namely Eoblattodea, which are characterized by long ovipositors. We speculate that the extinction of certain gymnosperm hosts almost ended the 200-My triumph of Eoblattodea. Despite an attempt to adapt to angiosperm hosts, Ensiferoblatta, Proceroblatta and suchlike cockroaches as an evolutionary dead end failed to save Eoblattodea from extinction. The lack of protection for eggs (maternal care in particular) might accelerate the extinction of Eoblattodea as a whole. creator: Xin-Ran Li creator: Di-Ying Huang uri: https://doi.org/10.7717/peerj.15067 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Li and Huang title: Seasonal and longitudinal water quality dynamics in three effluent-dependent rivers in Arizona link: https://peerj.com/articles/15069 last-modified: 2023-03-29 description: Effluent-fed streams, which receive inputs from wastewater treatment plants, are becoming increasingly common across the globe as urbanization intensifies. In semi-arid and arid regions, where many natural streams have dried up due to over extraction of water, many streams rely completely on treated effluent to sustain baseflow during dry seasons. These systems are often thought of as ‘second-class’ or highly disturbed stream ecosystems, but they have the potential to serve as refuges for native aquatic biota if water quality is high, especially in areas where few natural habitats remain. In this study, we investigated seasonal and longitudinal water quality dynamics at multiple sites across six reaches of three effluent-dependent rivers in Arizona (USA) with the objective (1) to quantify changes in effluent water quality due to distance traveled and season/climate and (2) to qualify whether water quality conditions in these systems are sufficient to support native aquatic species. Study reaches ranged in length from 3 to 31 km and in geographic setting from low desert to montane conifer forest. We observed the lowest water quality conditions (e.g., elevated temperature and low dissolved oxygen) during the summer in low desert reaches, and significantly greater natural remediation of water quality in longer vs. shorter reaches for several factors, including temperature, dissolved oxygen and ammonia. Nearly all sites met or exceeded water quality conditions needed to support robust assemblages of native species across multiple seasons. However, our results also indicated that temperature (max 34.2 °C), oxygen levels (min 2.7 mg/L) and ammonia concentrations (max 5.36 mg/L N) may occasionally be stressful for sensitive taxa at sites closest to effluent outfalls. Water quality conditions may be a concern during the summer. Overall, effluent-dependent streams have the capacity to serve as refuges for native biota in Arizona, and they may become the only aquatic habitat available in many urbanizing arid and semi-arid regions. creator: Hamdhani Hamdhani creator: Drew E. Eppehimer creator: David M. Quanrud creator: Michael T. Bogan uri: https://doi.org/10.7717/peerj.15069 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Hamdhani et al. title: A 3D-printed passive exoskeleton for upper limb assistance in children with motor disorders: proof of concept through an electromyography-based assessment link: https://peerj.com/articles/15095 last-modified: 2023-03-29 description: The rehabilitation of children with motor disorders is mainly focused on physical interventions. Numerous studies have demonstrated the benefits of upper function using robotic exoskeletons. However, there is still a gap between research and clinical practice, owing to the cost and complexity of these devices. This study presents a proof of concept of a 3D-printed exoskeleton for the upper limb, following a design that replicates the main characteristics of other effective exoskeletons described in the literature. 3D printing enables rapid prototyping, low cost, and easy adjustment to the patient anthropometry. The 3D-printed exoskeleton, called POWERUP, assists the user’s movement by reducing the effect of gravity, thereby allowing them to perform upper limb exercises. To validate the design, this study performed an electromyography-based assessment of the assistive performance of POWERUP, focusing on the muscular response of both the biceps and triceps during elbow flexion–extension movements in 11 healthy children. The Muscle Activity Distribution (MAD) is the proposed metric for the assessment. The results show that (1) the exoskeleton correctly assists elbow flexion, and (2) the proposed metric easily identifies the exoskeleton configuration: statistically significant differences (p-value = 2.26 ⋅ 10−7 < 0.001) and a large effect size (Cohen’s d = 3.78 > 0.8) in the mean MAD value were identified for both the biceps and triceps when comparing the transparent mode (no assistance provided) with the assistive mode (anti-gravity effect). Therefore, this metric was proposed as a method for assessing the assistive performance of exoskeletons. Further research is required to determine its usefulness for both the evaluation of selective motor control (SMC) and the impact of robot-assisted therapies. creator: Cristina Sanchez creator: Laura Blanco creator: Carmina del Río creator: Eloy Urendes creator: Vanina Costa creator: Rafael Raya uri: https://doi.org/10.7717/peerj.15095 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Sanchez et al. title: “Integrative learning” promotes learning but not memory in older rats link: https://peerj.com/articles/15101 last-modified: 2023-03-29 description: BackgroundWe had previously advanced the concept of “Integrative Learning”, that is, “under the role of ‘meta-learning self’, learners actively integrate learning materials to achieve rapid and in-depth understanding of knowledge”, and designed an animal behavioral model to compare the effects of “Integrative Learning” (IL) vs. “Progressive Learning” (PL) in young rats. It was found that IL is more advantageous than PL. Here, we aim to examine whether the same phenomenon persist in older rats.MethodsFifteen 12-month-old male Sprague-Dawley (SD) rats were selected as subjects and randomly divided into the IL group and the PL group, and a 14-unit integrative T-maze was constructed for the study. Training and testing procedures contained three stages: the learning stage, the memory retention test stage and the Gestalt transfer learning stage. Data on young rats (1-month-old) from the previous study were also drawn here for comparisons on learning performance.Results(1) The 12-session learning stage can be divided into three sub-stages as each sub-stage represented the new opening of one third of the whole path in the PL group. There were significant interactions in total errors made between groups and sessions: the PL group had significantly fewer errors during Sub-stage One due to a much shorter path to be learned, however, the IL group’s errors made sharply dropped as learning progressed into Sub-stage Two and Three, and were maintained at a significantly lower level than the PL group during Sub-stage Three. (2) When compared with young rats, age had a main effect on the number of errors made—the 1-month-old groups learned overall better and faster than the older groups, whereas the pattern of group differences between the IL and PL learning modes remained consistent across young and older groups. (3) Unlike young rats, during the memory retention test stage and the Gestalt transfer learning stage, the IL group did not perform better than the PL group in older rats.Conclusions(1) “Integrative Learning” promotes learning but not memory in older rats. (2) Higher-order cognitive abilities that support meta-cognition, long-term retention and knowledge transfer might be deteriorating in older rats. creator: Bin Yin creator: Xiao-Rui Wu creator: Rong Lian uri: https://doi.org/10.7717/peerj.15101 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Yin et al. title: Volcanic-associated ecosystems of the Mediterranean Sea: a systematic map and an interactive tool to support their conservation link: https://peerj.com/articles/15162 last-modified: 2023-03-29 description: BackgroundHydrothermal vents, cold seeps, pockmarks and seamounts are widely distributed on the ocean floor. Over the last fifty years, the knowledge about these volcanic-associated marine ecosystems has notably increased, yet available information is still limited, scattered, and unsuitable to support decision-making processes for the conservation and management of the marine environment.MethodsHere we searched the Scopus database and the platform Web of Science to collect the scientific information available for these ecosystems in the Mediterranean Sea. The collected literature and the bio-geographic and population variables extracted are provided into a systematic map as an online tool that includes an updated database searchable through a user-friendly R-shiny app.ResultsThe 433 literature items with almost one thousand observations provided evidence of more than 100 different volcanic-associated marine ecosystem sites, mostly distributed in the shallow waters of the Mediterranean Sea. Less than 30% of these sites are currently included in protected or regulated areas. The updated database available in the R-shiny app is a tool that could guide the implementation of more effective protection measures for volcanic-associated marine ecosystems in the Mediterranean Sea within existing management instruments under the EU Habitats Directive. Moreover, the information provided in this study could aid policymakers in defining the priorities for the future protection measures needed to achieve the targets of the UN Agenda 2030. creator: Valentina Costa creator: Valentina Sciutteri creator: Pierpaolo Consoli creator: Elisabetta Manea creator: Elisabetta Menini creator: Franco Andaloro creator: Teresa Romeo creator: Roberto Danovaro uri: https://doi.org/10.7717/peerj.15162 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Costa et al. title: Micro-shear bond strength of different calcium silicate materials to bulk-fill composite link: https://peerj.com/articles/15183 last-modified: 2023-03-29 description: IntroductionThis study aimed to compare the micro-shear bond strength (µSBS) performances of two resin-based calcium silicate-based cement (CSC) (TheraCal PT and TheraCal LC), Biodentine, and two modified-MTA CSC materials (NeoMTA 2 and BioMTA+) to bulk-fill restorative material.Materials and MethodsFifty 3D printed cylindrical resin blocks with a central hole were used (2 mm in depth and 4 mm in diameter). CSCs were placed in the holes (per each group n = 10) and incubated for 24 h. Cylindrical polyethylene molds (2 mm in height and diameter) were used to place the bulk-fill restorative materials on the CSCs and polymerize for 20 s. Then, all specimens were incubated for 24 h at 37 °C at a humidity of 100%. Specimen’s µSBSs were determined with a universal testing machine. Data were analyzed with one-way ANOVA (Welch) and Tamhane test.ResultsStatistically higher µSBS was found for TheraCal PT (29.91 ± 6.13 MPa) (p < 0.05) respect to all the other materials tested. TheraCal LC (20.23 ± 6.32 MPa) (p > 0.05) reported higher µSBS than NeoMTA 2 (11.49 ± 5.78 MPa) and BioMTA+ (6.45 ± 1.89 MPa) (p < 0.05). There was no statistical difference among TheraCal LC, NeoMTA 2 and Biodentine (15.23 ± 7.37 MPa) and between NeoMTA 2 and BioMTA+ (p > 0.05).ConclusionChoosing TheraCal PT as the pulp capping material may increase the adhesion and µSBS to the bulk-fill composite superstructure and sealing ability. creator: Seda Falakaloğlu creator: Merve Yeniçeri Özata creator: Gianluca Plotino uri: https://doi.org/10.7717/peerj.15183 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Falakaloğlu et al. title: The assessment of internal adaptation and fracture resistance of glass ionomer and resin-based restorative materials applied after different caries removal techniques in primary teeth: an in-vitro study link: https://peerj.com/articles/14825 last-modified: 2023-03-28 description: BackgroundThe aim of this study was to evaluate the 3-dimensional (3D) internal adaptation (IA) and fracture resistance (FR) of compomer and glass ionomers applied after conventional caries removal to sound dentin (CCRSD) and selective caries removal to firm dentin (SCRFD) in in-vitro.MethodsThirty extracted primary molars were randomly assigned to three main groups (n = 10) as glass hybrid restorative (GHR) (Equia Forte® HT), conventional glass ionomer (CGIR) (Voco Ionofil Molar) and compomer (Dyract XP). Each group was randomly divided into two subgroups according to caries removal technique as CCRSD (n = 5) and SCRFD (n = 5). The restoration procedures were completed after caries removal (CCRSD or SCRFD) in all samples. Then, specimens were subjected to IA and FR tests. Data were analyzed with Student’s t, one-way ANOVA, and Kruskal Wallis-H tests. The correlation between IA and FR results was analyzed with a Pearson test. The statistical significance level was considered as 5%.ResultsWhile CCRSD showed superior IA results than SCRFD for all restorative materials (p < 0.05), no statistical difference was found between CCRSD and SCRFD in FR assessment (p > 0.05). In CCRSD, compomer showed superior results for IA and FR than glass ionomers (p < 0.05). In SCRFD, it was found no significant difference between the restoratives for IA (p > 0.05). However, compomer showed superior FR results than glass ionomers (p < 0.05). There was moderate negative correlation between internal voids and FR without statistically significant difference (r = −0.333, p = 0.072).ConclusionsDespite the advantages of SCRFD, it was found to be less superior than CCRSD in IA assessment. Therefore, when SCRFD is preferred, a peripheral seal should be provided for ideal restorative treatment. On the other hand, compomer mostly showed superior results compared to others. creator: Akif Demirel creator: Ayşe Işıl Orhan creator: Arda Büyüksungur uri: https://doi.org/10.7717/peerj.14825 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Demirel et al. title: Pandemic-driven changes in the nearshore non-commercial fishery in Hawai’i: catch photos posted to social media capture changes in fisher behavior link: https://peerj.com/articles/14994 last-modified: 2023-03-28 description: Using social media, we collect evidence for how nearshore fisheries are impacted by the global COVID-19 pandemic in Hawai’i. We later confirm our social media findings and obtain a more complete understanding of the changes in nearshore non-commercial fisheries in Hawai’i through a more conventional approach—speaking directly with fishers. Resource users posted photographs to social media nearly three times as often during the pandemic with nearly double the number of fishes pictured per post. Individuals who fished for subsistence were more likely to increase the amount of time spent fishing and relied more on their catch for food security. Furthermore, individuals fishing exclusively for subsistence were more likely to fish for different species during the pandemic than individuals fishing recreationally. Traditional data collection methods are resource-intensive and this study shows that during times of rapid changes, be it ecological or societal, social media can more quickly identify how near shore marine resource use adapts. As climate change threatens additional economic and societal disturbances, it will be necessary for resource managers to collect reliable data efficiently to better target monitoring and management efforts. creator: Timothy Grabowski creator: Michelle E. Benedum creator: Andrew Curley creator: Cole Dill-De Sa creator: Michelle Shuey uri: https://doi.org/10.7717/peerj.14994 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Grabowski et al. title: Identification and biodiversity patterns of Aspergillus species isolated from some soil invertebrates at high altitude using morphological characteristics and phylogenetic analyses link: https://peerj.com/articles/15035 last-modified: 2023-03-28 description: BackgroundThe carcinogenic, mutagenic, and teratogenic chemicals such as aflatoxin are a worldwide health problem. Aspergillus spp., responsible for most cases of aflatoxin contamination, are common in the environment and spread easily to many different types of food. The objectives of this study were to conduct a survey of fungi associated with three soil invertebrates in Taif, Saudi Arabia, identify these isolates and explore mycotoxins formation.MethodsIn total, 114 fungal isolates were collected from various soil invertebrates (millipedes, Armadillidium vulgare and Porcellio laevis) in Taif, Saudi Arabia, among them, 22 isolates were identified as Aspergillus spp. based on morphological and molecular characteristics followed by both Fusarium and Penicillium.ResultsThe sequences of ITS 1 and ITS 4 were utilized. Using bootstrap analysis, phylogenetic tree was split into two distinct clusters. Five sub clusters were included inside the first major cluster, and their bootstrap value was 99%. While, there were two small clusters in the second major cluster. All the tested Aspergillus strains were able to have a single PCR fragment amplified using the primer AspTef. TEF-1 DNA sequence bootstrap analysis with 1,000 replicates revealed two distinct groups. Additionally, the Aspergillus isolates were grouped into two different clusters with about 65% genetic similarity using ISSR-PCR analysis. The standard polymerase chain reaction was used to effectively amplify the Aopks, afl-A and omt-A genes in aflatoxigenic Aspergillus strains. Four Aspergillus strains used in this investigation were shown to generate aflatoxin B1. While, three Aspergillus stains showed ochratoxin genes.ConclusionsIn conclusion, the results indicate significant differences in the fungal community between ecoregions and soil invertebrates. Moreover, mycotoxin detection and identification among Aspergillus isolates were elucidated. This study could shed light on the risk of mycotoxin contamination along the supply chain. creator: Mohamed Fadl Awad creator: Bander Albogami creator: Tarombera Mwabvu creator: Montaser M. Hassan creator: Alaa Baazeem creator: Mohamed M. Hassan creator: Mohsen Mohamed Elsharkawy uri: https://doi.org/10.7717/peerj.15035 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Awad et al. title: Sex impacts pain behaviour but not emotional reactivity of lambs following ring tail docking link: https://peerj.com/articles/15092 last-modified: 2023-03-28 description: Studies in humans have shown sex differences in response to painful events, however, little is known in relation to sex differences in sheep. Understanding sex differences would enable improved experimental design and interpretation of studies of painful procedures in sheep. To examine sex differences in response to pain, 80 lambs were tested across five cohorts of 16. The lambs were penned in groups containing two male and two female lambs with their respective mothers. Lambs were randomly allocated from within each block to one of four treatment groups; FRing–Female lamb, ring tail docked without analgesia, MRing–Male lamb, ring tail docked without analgesia, FSham–Female lamb, tail manipulated and MSham–Male lamb, tail manipulated. Following treatment, lambs were returned to their pen and were video recorded for 45 mins for behavioural observations of acute pain and posture. An hour after treatment, lambs then underwent an emotional reactivity test that consisted of three phases: Isolation, Novelty and Startle. Following treatment, Ring lambs displayed more abnormal postures (mean = 2.5 ± 0.5) compared to Sham lambs (mean = 0.05 ± 0.4, P = 0.0001). There was an effect of sex on the display of acute pain-related behaviours in lambs that were tail docked (P < 0.001), with female lambs displaying more acute behaviours (mean count = +2.2). This difference in behaviour between sexes was not observed in Sham lambs. There was no effect of sex on display of postures related to pain (P = 0.99). During the Novelty and Startle phase of the emotional reactivity test, Ring lambs tended to (P = 0.084) or did (P = 0.018) show more fear related behaviours, respectively. However, no effect of sex was observed. The results of this study indicate that a pain state may alter the emotional response of lambs to novel objects and potential fearful situations. It was also demonstrated that female lambs display increased sensitivity to the acute pain caused by tail docking compared to males. creator: Danila Marini creator: Jessica E. Monk creator: Dana L.M. Campbell creator: Caroline Lee creator: Sue Belson creator: Alison Small uri: https://doi.org/10.7717/peerj.15092 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Marini et al. title: Variations and gradients between methane seep and off-seep microbial communities in a submarine canyon system in the Northeast Pacific link: https://peerj.com/articles/15119 last-modified: 2023-03-28 description: Methane seeps are highly abundant marine habitats that contribute sources of chemosynthetic primary production to marine ecosystems. Seeps also factor into the global budget of methane, a potent greenhouse gas. Because of these factors, methane seeps influence not only local ocean ecology, but also biogeochemical cycles on a greater scale. Methane seeps host specialized microbial communities that vary significantly based on geography, seep gross morphology, biogeochemistry, and a diversity of other ecological factors including cross-domain species interactions. In this study, we collected sediment cores from six seep and non-seep locations from Grays and Quinault Canyons (46–47°N) off Washington State, USA, as well as one non-seep site off the coast of Oregon, USA (45°N) to quantify the scale of seep influence on biodiversity within marine habitats. These samples were profiled using 16S rRNA gene sequencing. Predicted gene functions were generated using the program PICRUSt2, and the community composition and predicted functions were compared among samples. The microbial communities at seeps varied by seep morphology and habitat, whereas the microbial communities at non-seep sites varied by water depth. Microbial community composition and predicted gene function clearly transitioned from on-seep to off-seep in samples collected from transects moving away from seeps, with a clear ecotone and high diversity where methane-fueled habitats transition into the non-seep deep sea. Our work demonstrates the microbial and metabolic sphere of influence that extends outwards from methane seep habitats. creator: Milo E Cummings creator: Lila M. Ardor Bellucci creator: Sarah Seabrook creator: Nicole A. Raineault creator: Kerry L. McPhail creator: Andrew R. Thurber uri: https://doi.org/10.7717/peerj.15119 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Cummings et al. title: The modulatory effects of gut microbes and metabolites on blood–brain barrier integrity and brain function in sepsis-associated encephalopathy link: https://peerj.com/articles/15122 last-modified: 2023-03-28 description: BackgroundIntestinal microbiota homeostasis and the gut-brain axis are key players associated with host health and alterations in metabolic, inflammatory, and neurodegenerative disorders. Sepsis-associated encephalopathy (SAE), which is closely associated with bacterial translocation, is a common secondary organ dysfunction and an urgent, unsolved problem affecting patient quality of life. Our study examined the neuroprotective effects of the gut microbiome and short-chain fatty acid (SCFA) metabolites on SAE.MethodsMale C57BL/6 mice were administered SCFAs in drinking water, then subjected to cecal ligation and puncture (CLP) surgery to induce SAE. 16S rRNA sequencing was used to investigate gut microbiome changes. The open field test (OFT) and Y-maze were performed to evaluate brain function. The permeability of the blood–brain barrier (BBB) was assessed by Evans blue (EB) staining. Hematoxylin and eosin (HE) staining was used to examine intestinal tissue morphology. The expression levels of tight junction (TJ) proteins and inflammatory cytokines was assessed by western blots and immunohistochemistry. In vitro, bEND.3 cells were incubated with SCFAs and then with lipopolysaccharide (LPS). Immunofluorescence was used to examine the expression of TJ proteins.ResultsThe composition of the gut microbiota was altered in SAE mice; this change may be related to SCFA metabolism. SCFA treatment significantly alleviated behavioral dysfunction and neuroinflammation in SAE mice. SCFAs upregulated occludin and ZO-1 expression in the intestine and brain in SAE mice and LPS-treated cerebromicrovascular cells.ConclusionsThese findings suggested that disturbances in the gut microbiota and SCFA metabolites play key roles in SAE. SCFA supplementation could exert neuroprotective effects against SAE by preserving BBB integrity. creator: Zhaoying Li creator: Fangxiang Zhang creator: Meisha Sun creator: Jia Liu creator: Li Zhao creator: Shuchun Liu creator: Shanshan Li creator: Bin Wang uri: https://doi.org/10.7717/peerj.15122 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Li et al. title: Day and night camera trap videos are effective for identifying individual wild Asian elephants link: https://peerj.com/articles/15130 last-modified: 2023-03-28 description: Regular monitoring of wild animal populations through the collection of behavioral and demographic data is critical for the conservation of endangered species. Identifying individual Asian elephants (Elephas maximus), for example, can contribute to our understanding of their social dynamics and foraging behavior, as well as to human-elephant conflict mitigation strategies that account for the behavior of specific individuals involved in the conflict. Wild elephants can be distinguished using a variety of different morphological traits—e.g., variations in ear and tail morphology, body scars and tumors, and tusk presence, shape, and length—with previous studies identifying elephants via direct observation or photographs taken from vehicles. When elephants live in dense forests like in Thailand, remote sensing photography can be a productive approach to capturing anatomical and behavioral information about local elephant populations. While camera trapping has been used previously to identify elephants, here we present a detailed methodology for systematic, experimenter differentiation of individual elephants using data captured from remote sensing video camera traps. In this study, we used day and night video footage collected remotely in the Salakpra Wildlife Sanctuary in Thailand and identified 24 morphological characteristics that can be used to recognize individual elephants. A total of 34 camera traps were installed within the sanctuary as well as crop fields along its periphery, and 107 Asian elephants were identified: 72 adults, 11 sub-adults, 20 juveniles, and four infants. We predicted that camera traps would provide enough information such that classified morphological traits would aid in reliably identifying the adult individuals with a low probability of misidentification. The results indicated that there were low probabilities of misidentification between adult elephants in the population using camera traps, similar to probabilities obtained by other researchers using handheld cameras. This study suggests that the use of day and night video camera trapping can be an important tool for the long-term monitoring of wild Asian elephant behavior, especially in habitats where direct observations may be difficult. creator: Sasha Montero-De La Torre creator: Sarah L. Jacobson creator: Martin Chodorow creator: Marnoch Yindee creator: Joshua M. Plotnik uri: https://doi.org/10.7717/peerj.15130 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Montero-De La Torre et al. title: Morphological, ultrastructural and molecular variations in susceptible and resistant genotypes of chickpea infected with Botrytis grey mould link: https://peerj.com/articles/15134 last-modified: 2023-03-28 description: Biotic stress due to fungal infection is detrimental to the growth and development of chickpea. In our study, two chickpea genotypes vizCicer pinnatifidum (resistant) and PBG5 (susceptible) were inoculated with (1 × 104 spore mL−1) of nectrotrophic fungus Botrytis cinerea at seedling stage. These seedlings were evaluated for morphological, ultrastructural, and molecular differences after 3, 5 and 7 days post inoculation (dpi). Visual symptoms were recorded in terms of water-soaked lesions, rotten pods and twigs with fungal colonies. Light and scanning electron microscopy (SEM) revealed the differences in number of stomata, hyphal network and extent of topographical damage in resistant (C. pinnatifidum) and susceptible (PBG5) genotypes, which were validated by stomatal index studies done by using fluorescence microscopy in the infection process of B. cinerea in leaves of both chickpea genotypes. In case of control (water inoculated) samples, there were differences in PCR analysis done using five primers for screening the genetic variations between two genotypes. The presence of a Botrytis responsive gene (LrWRKY) of size ~300 bp was observed in uninoculated resistant genotype which might have a role in resistance against Botrytis grey mould. The present investigation provides information about the variation in the infection process of B. cinerea in two genotypes which can be further exploited to develop robust and effective strategies to manage grey mould disease. creator: Richa Thakur creator: Rajni Devi creator: Milan Kumar Lal creator: Rahul Kumar Tiwari creator: Sucheta Sharma creator: Ravinder Kumar uri: https://doi.org/10.7717/peerj.15134 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Thakur et al. title: High expression of six-transmembrane epithelial antigen of prostate 3 promotes the migration and invasion and predicts unfavorable prognosis in glioma link: https://peerj.com/articles/15136 last-modified: 2023-03-28 description: Recent studies have suggested that ferroptosis, a form of iron-dependent regulated cell death, might play essential roles in tumor initiation and progression. Six-transmembrane epithelial antigen of prostate 3 (STEAP3) is a ferrireductase involved in the regulation of intracellular iron homeostasis. However, the clinical significance and biological function of STEAP3 in human cancers remain poorly understood. Through a comprehensive bioinformatics analysis, we found that STEAP3 mRNA and protein expression were up-regulated in GBM, LUAD, and UCEC, and down-regulated in LIHC. Survival analysis indicated that STEAP3 had prognostic significance only in glioma. Multivariate Cox regression analysis revealed that high STEPA3 expression was correlated with poor prognosis. STEAP3 expression was significantly negatively correlated with promoter methylation level, and patients with lower STEAP3 methylation level had worse prognosis than those with higher STEAP3 methylation level. Single-cell functional state atlas showed that STEAP3 regulated epithelial-to-mesenchymal transition (EMT) in GBM. Furthermore, the results of wound healing and transwell invasion assays demonstrated that knocking down STEAP3 inhibited the migration and invasion of T98G and U251 cells. Functional enrichment analysis suggested that genes co-expressed with STEAP3 mainly participated in inflammation and immune-related pathways. Immunological analysis revealed that STEAP3 expression was significantly correlated with immune infiltration cells, including macrophages and neutrophils, especially the M2 macrophages. Individuals with low STEAP3 expression were more likely to respond to immunotherapy than those with high STEAP3 expression. These results suggest that STEAP3 promotes glioma progression and highlight its pivotal role in regulating immune microenvironment. creator: Langmei Deng creator: Shuangshuang Zeng creator: Qiaoli Yi creator: Liying Song uri: https://doi.org/10.7717/peerj.15136 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Deng et al. title: Functional characterization of the GhNRT2.1e gene reveals its significant role in improving nitrogen use efficiency in Gossypium hirsutum link: https://peerj.com/articles/15152 last-modified: 2023-03-28 description: BackgroundNitrate is the primary type of nitrogen available to plants, which is absorbed and transported by nitrate transporter 2 (NRT2) at low nitrate conditions.MethodsGenome-wide identification of NRT2 genes in G. hirsutum was performed. Gene expression patterns were revealed using RNA-seq and qRT-PCR. Gene functions were characterized using overexpression in A. thaliana and silencing in G. hirsutum. Protein interactions were verified by yeast two-hybrid and luciferase complementation imaging (LCI) assays.ResultsWe identified 14, 14, seven, and seven NRT2 proteins in G. hirsutum, G. barbadense, G. raimondii, and G. arboreum. Most NRT2 proteins were predicted in the plasma membrane. The NRT2 genes were classified into four distinct groups through evolutionary relationships, with members of the same group similar in conserved motifs and gene structure. The promoter regions of NRT2 genes included many elements related to growth regulation, phytohormones, and abiotic stresses. Tissue expression pattern results revealed that most GhNRT2 genes were specifically expressed in roots. Under low nitrate conditions, GhNRT2 genes exhibited different expression levels, with GhNRT2.1e being the most up-regulated. Arabidopsis plants overexpressing GhNRT2.1e exhibited increased biomass, nitrogen and nitrate accumulation, nitrogen uptake and utilization efficiency, nitrogen-metabolizing enzyme activity, and amino acid content under low nitrate conditions. In addition, GhNRT2.1e-silenced plants exhibited suppressed nitrate uptake and accumulation, hampered plant growth, affected nitrogen metabolism processes, and reduced tolerance to low nitrate. The results showed that GhNRT2.1e could promote nitrate uptake and transport under low nitrate conditions, thus effectively increasing nitrogen use efficiency (NUE). We found that GhNRT2.1e interacts with GhNAR2.1 by yeast two-hybrid and LCI assays.DiscussionOur research lays the foundation to increase NUE and cultivate new cotton varieties with efficient nitrogen use. creator: Xinmiao Zhang creator: Jiajia Feng creator: Ruolin Zhao creator: Hailiang Cheng creator: Javaria Ashraf creator: Qiaolian Wang creator: Limin Lv creator: Youping Zhang creator: Guoli Song creator: Dongyun Zuo uri: https://doi.org/10.7717/peerj.15152 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Zhang et al. title: Plant-exuded chemical signals induce surface attachment of the bacterial pathogen Pseudomonas syringae link: https://peerj.com/articles/14862 last-modified: 2023-03-27 description: Many plant pathogenic bacteria suppress host defenses by secreting small molecule toxins or immune-suppressing proteins into host cells, processes that likely require close physical contact between pathogen and host. Yet, in most cases, little is known about whether phytopathogenic bacteria physically attach to host surfaces during infection. Here we report that Pseudomonas syringae pv. tomato strain DC3000, a Gram-negative bacterial pathogen of tomato and Arabidopsis, attaches to polystyrene and glass surfaces in response to chemical signals exuded from Arabidopsis seedlings and tomato leaves. We characterized the molecular nature of these attachment-inducing signals and discovered that multiple hydrophilic metabolites found in plant exudates, including citric acid, glutamic acid, and aspartic acid, are potent inducers of surface attachment. These same compounds were previously identified as inducers of P. syringae genes encoding a type III secretion system (T3SS), indicating that both attachment and T3SS deployment are induced by the same plant signals. To test if surface attachment and T3SS are regulated by the same signaling pathways, we assessed the attachment phenotypes of several previously characterized DC3000 mutants, and found that the T3SS master regulator HrpL was partially required for maximal levels of surface attachment, whereas the response regulator GacA, a negative regulator of T3SS, negatively regulated DC3000 surface attachment. Together, our data indicate that T3SS deployment and surface attachment by P. syringae may be co-regulated by the same host signals during infection, possibly to ensure close contact necessary to facilitate delivery of T3SS effectors into host cells. creator: Megan R. O’Malley creator: Eyram Kpenu creator: Scott C. Peck creator: Jeffrey C. Anderson uri: https://doi.org/10.7717/peerj.14862 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 O’Malley et al. title: m6A reader IGF2BP1 accelerates apoptosis of high glucose-induced vascular endothelial cells in a m6A-HMGB1 dependent manner link: https://peerj.com/articles/14954 last-modified: 2023-03-27 description: Emerging evidence indicates that N6-methyladenosine (m6A) plays a critical role in vascular biological characteristic. In diabetes mellitus pathophysiology, high glucose (HG)-induced vascular endothelial dysfunction is associated with diabetes vascular complications. Nevertheless, the underlying mechanism of high glucose (HG)-related m6A regulation on vascular endothelial cells is still unclear. Results indicated that m6A reader insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) was up-regulated in HG-treated human umbilical vascular endothelium cells (HUVECs) comparing to normal group. Functionally, results indicated that IGF2BP1 knockdown recovered the proliferation of HUVECs inhibited by HG-administration. Besides, IGF2BP1 knockdown reduced the apoptosis induced by HG-administration. Mechanistically, IGF2BP1 interacted with HMGB1 mRNA and stabilized its expression of m6A-modified RNA. Therefore, these findings provided compelling evidence demonstrating that m6A reader IGF2BP1 contributes to the proliferation and apoptosis of vascular endothelial cells in hyperglycaemia, serving as a target for development of diabetic angiopathy therapeutics. creator: Anru Liang creator: Jianyu Liu creator: Yanlin Wei creator: Yuan Liao creator: Fangxiao Wu creator: Jiang Ruan creator: Junjun Li uri: https://doi.org/10.7717/peerj.14954 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Liang et al. title: Genomic assessment reveals signal of adaptive selection in populations of the Spotted rose snapper Lutjanus guttatus from the Tropical Eastern Pacific link: https://peerj.com/articles/15029 last-modified: 2023-03-27 description: BackgroundThe lack of barriers in the marine environment has promoted the idea of panmixia in marine organisms. However, oceanographic conditions and habitat characteristics have recently been linked to genetic structure in marine species. The Tropical Eastern Pacific (TEP) is characterized by dynamic current systems and heterogeneous oceanographic conditions. The Gulf of Panama (part of the equatorial segment for the TEP) is influenced by a complex current system and heterogeneous environment, which has been shown to limit the gene flow for shoreline species. Next Generation Sequencing (NGS) has contributed to detect genetic differences in previously reported panmictic species by the assessment of loci associated with selection and to understand how selection acts affects marine populations. Lutjanus guttatus is a species distributed in the TEP for which previous studies using mitochondrial data recovered a panmictic pattern along its distributional range. In this study, we used SNP data of L. guttatus individuals sampled along its range to evaluate population genetic structure and investigate whether oceanographic factors influence the species’ genetic architecture. Finally, we assessed the role of adaptive selection by evaluating the contribution of outlier and neutral loci to genetic divergence.MethodsThe RADcap method was used to obtain 24 million paired reads for 123 individuals of L. guttatus covering nearly all its distributional area. Genetic variation was assessed using both spatial and non-spatial methods by comparing three different data sets: (i) a Combined Loci (CL dataset = 2003 SNPs); a search for putative loci under selection allowed the evaluation of (ii) Neutral Loci (NL dataset = 1858 SNPs) and (iii) Outlier Loci (OL dataset = 145 SNPs). We used the estimating effective migration surface (EEMS) approach to detect possible barriers to gene flow.ResultsGenetic differences were found in the OL dataset, showing two clusters (Northern and Southern), whereas NL showed no differences. This result may be related to the Selection-Migration balance model. The limit between the Northern and Southern groups was in the Gulf of Panama, which has been previously identified as a barrier to gene flow for other species, mainly due to its heterogeneous oceanographic conditions. The results suggest that selection plays an important role in generating genetic differences in Lutjanus guttatus. A migration corridor was detected that coincides with the Costa Rica Coastal Current that flows from Central America to the Gulf of California, allowing the homogenization of the northern population. In the Southern cluster, a migration corridor was observed with the OL from Panama to Colombia, which could be associated with the currents found in the Gulf of Panama. Genetic variation found in the OL of Lutjanus guttatus highlights the usefulness of NGS data in evaluating the role of selection in population differentiation. creator: Adán F. Mar-Silva creator: Pindaro Diaz-Jaimes creator: Cristina Domínguez-Mendoza creator: Omar Domínguez-Domínguez creator: Jonathan Valdiviezo-Rivera creator: Eduardo Espinoza-Herrera uri: https://doi.org/10.7717/peerj.15029 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Mar-Silva et al. title: A clinical medicine level test at Jinan University School of Medicine reveals the importance of training medical students in clinical history-taking link: https://peerj.com/articles/15052 last-modified: 2023-03-27 description: BackgroundsTraining in the basic interview skills of clinical history-taking has always been a significant component of medical education.PurposeThis study was designed to identify the factors influence medical students’ history-taking skills learning and develop a way to improve these skills.MethodsWe firstly analysed the academic performance of medical students at Jinan University School of Medicine in different disciplines of the Clinical Medicine Level Test (CMLT), to ensure the students have obtained comprehensive medical education prior to beginning their clinical internships. Next, we conducted a survey among the CMLT participants to seek the underlying causes and corresponding measures to improve history-taking in the future. Before these medical students entered their fifth-year clinical practice, we finally provide them with pre-internship training, including the history-taking workshops with standard patients (SP).ResultsThe analysis of the clinical skill sections of the CMLT revealed that the students performed significantly better on clinical operations from multiple disciplines than on medical history-taking. Principal component analysis of the survey questionnaire indicated that the capability of history-taking, course assessments, and awareness of the value of medical history-taking emerged as the key factors forming a cohesive clue for sustaining history-taking implementation. The intervention workshops of employing SP had a positive impact, as evidenced by the students’ feedback and suggestions for improving their ability of history-taking.ConclusionsThis study suggests that strengthening of medical history-taking training is indispensable for training qualified medical students. Workshops with SP is a successful teaching strategy for practicing history-taking, allowing students to spot minute errors and training communication skills. creator: Xianjun Meng creator: Mingya Zhang creator: Wei Ma creator: Xin Cheng creator: Xuesong Yang uri: https://doi.org/10.7717/peerj.15052 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Meng et al. title: Seed priming with essential oils for sustainable wheat agriculture in semi-arid region link: https://peerj.com/articles/15126 last-modified: 2023-03-27 description: Drought is one of the major constraints to global crop production. A number of sustainable systems have focused on the development of environmentally friendly innovative biotechnological interventions to prevent yield losses. The use of essential oils as a seed priming agent can make an important contribution as a natural stimulant in increasing drought stress tolerance. This study focuses on the effects of seeds coated with different doses (D0 (0%), D1 (0.01%), D2 (0.05%), D3 (0.10%) and D4 (0.25%)) of sage, rosemary and lavender essential oils on wheat germination, seedling establishment and yield parameters. Turkey’s local wheat genotype Köse was used as plant material. The impact of the seed priming on germination rate, coleoptile length, shoot length, root length, shoot fresh and dry weight, root fresh and dry weight, relative water content (RWC), proline, and chlorophyll contents was assessed in laboratory experiments. In addition, the effect of essential oil types on yield parameters and agronomic components (plant height, spike height, number of grains per spike, grain yield per spike, grain yield per unit area, thousand-grain weight) was evaluated in a field experiment during the 2019–2020 crop seasons in a semi-arid climate. According to laboratory results, the highest germination rate among all treatment doses was determined in the D2 treatment (rosemary 93.30%, sage 94.00% and lavender 92.50%), while the lowest germination rates for all essential oil types were determined in the D4 treatment (rosemary 41.70%, sage 40.90% and lavender 40.90%). Increasing treatment doses showed a similar suppressive effect on the other parameters. In the field experiment, the highest grain yield (256.52 kg/da) and thousand-grain weight (43.30 g) were determined in the rosemary treatment. However, the priming treatment has an insignificant on the number of grains per spike and the spike length. The light of these results, the effects of essential oil types and doses on yield parameters were discussed. The findings highlight the importance of using essential oils in seed priming methods for sustainable agricultural practices. creator: Muhammet Çağrı Oğuz creator: Ezgi Oğuz creator: Mustafa Güler uri: https://doi.org/10.7717/peerj.15126 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Oğuz et al. title: Reconstructing the dietary habits and trophic positions of the Longipterygidae (Aves: Enantiornithes) using neontological and comparative morphological methods link: https://peerj.com/articles/15139 last-modified: 2023-03-27 description: The Longipterygidae are a unique clade among the enantiornithines in that they exhibit elongate rostra (≥60% total skull length) with dentition restricted to the distal tip of the rostrum, and pedal morphologies suited for an arboreal lifestyle (as in other enantiornithines). This suite of features has made interpretations of this group’s diet and ecology difficult to determine due to the lack of analogous taxa that exhibit similar morphologies together. Many extant bird groups exhibit rostral elongation, which is associated with several disparate ecologies and diets (e.g., aerial insectivory, piscivory, terrestrial carnivory). Thus, the presence of rostral elongation in the Longipterygidae only somewhat refines trophic predictions of this clade. Anatomical morphologies do not function singularly but as part of a whole and thus, any dietary or ecological hypothesis regarding this clade must also consider other features such as their unique dentition. The only extant group of dentulous volant tetrapods are the chiropterans, in which tooth morphology and enamel thickness vary depending upon food preference. Drawing inferences from both avian bill proportions and variations in the dental morphology of extinct and extant taxa, we provide quantitative data to support the hypothesis that the Longipterygidae were animalivorous, with greater support for insectivory. creator: Alexander D. Clark creator: Han Hu creator: Roger BJ Benson creator: Jingmai K. O’Connor uri: https://doi.org/10.7717/peerj.15139 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Clark et al. title: Association between ACE and ACTN3 genes polymorphisms and athletic performance in elite and sub-elite Chinese youth male football players link: https://peerj.com/articles/14893 last-modified: 2023-03-24 description: BackgroundPrevious studies have shown controversial relationships between ACE I/D and ACTN3 R577x polymorphisms and athletic performance. Therefore, the aim of this study was to assess athletic performance indicators of Chinese youth male football players with different ACE and ACTN3 gene profiles.Methods and MaterialsThis study recruited 73 elite (26 13-year-olds, 28 14-year-olds, and 19 15-year-olds) and 69 sub-elite (37 13-year-olds, 19 14-year-olds, and 13 15-year-olds) and 107 controls (63 13-year-olds, and 44 14-year olds aged 13–15 years, all participants were of Chinese Han origin. We measured height, body mass, thigh circumference, speed, explosive power, repeat sprints ability, and aerobic endurance in elite and sub-elite players. We used single nucleotide polymorphism technology to detect controls elite and sub-elite players’ ACE and ACTN3 genotypes, Chi-squared (χ2) tests were employed to test for Hardy-Weinberg equilibrium. χ2 tests were also used to observe the association between the genotype distribution and allele frequencies between controls and elite and sub-elite players. The differences in parameters between the groups were analyzed using one-way analysis of variance and a Bonferroni’s post-hoc test, with statistical significance set at p ≤ 0.05.Results(1) The genotype distribution of the ACE I/D and ACTN3 R577x polymorphisms in controls, elite and sub-elite football players were consistent with Hardy-Weinberg equilibrium, except for the ACE genotype distribution of sub-elite players. (2) The RR and DD genotypes were significantly different between elite and sub-elite players (p = 0.024 and p = 0.02, respectively). (3) Elite players were more likely to have the RR genotype and less likely to have the DD genotype compared with sub-elite players. (4) Both elite and sub-elite RR players’ Yo-yo intermittent recovery level 1 (YYIR1) running distance was significantly longer than that of RX players (p = 0.05 and p = 0.025, respectively). However, there was no significantly different in YYIR1 running distance between elite and sub-elite RR players. (5) Elite XX players’ VO2 max was significantly higher than that of RX and sub-elite players.ConclusionThese results indicate that ACE I/D and ACTN3 R577x polymorphisms are not associated with muscle power in Chinese elite and sub-elite players. The XX genotype of ACTN3 is associated with the aerobic endurance of elite players. creator: Shidong Yang creator: Wentao Lin creator: Mengmeng Jia creator: Haichun Chen uri: https://doi.org/10.7717/peerj.14893 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Yang et al. title: Economic costs of invasive rodents worldwide: the tip of the iceberg link: https://peerj.com/articles/14935 last-modified: 2023-03-24 description: BackgroundRodents are among the most notorious invasive alien species worldwide. These invaders have substantially impacted native ecosystems, food production and storage, local infrastructures, human health and well-being. However, the lack of standardized and understandable estimation of their impacts is a serious barrier to raising societal awareness, and hampers effective management interventions at relevant scales.MethodsHere, we assessed the economic costs of invasive alien rodents globally in order to help overcome these obstacles. For this purpose, we combined and analysed economic cost data from the InvaCost database—the most up-to-date and comprehensive synthesis of reported invasion costs—and specific complementary searches within and beyond the published literature.ResultsOur conservative analysis showed that reported costs of rodent invasions reached a conservative total of US$ 3.6 billion between 1930 and 2022 (annually US$ 87.5 million between 1980 and 2022), and were significantly increasing through time. The highest cost reported was for muskrat Ondatra zibethicus (US$ 377.5 million), then unspecified Rattus spp. (US$ 327.8 million), followed by Rattus norvegicus specifically (US$ 156.6 million) and Castor canadensis (US$ 150.4 million). Of the total costs, 87% were damage-related, principally impacting agriculture and predominantly reported in Asia (60%), Europe (19%) and North America (9%). Our study evidenced obvious cost underreporting with only 99 documents gathered globally, clear taxonomic gaps, reliability issues for cost assessment, and skewed breakdowns of costs among regions, sectors and contexts. As a consequence, these reported costs represent only a very small fraction of the expected true cost of rodent invasions (e.g., using a less conservative analytic approach would have led to a global amount more than 80-times higher than estimated here).ConclusionsThese findings strongly suggest that available information represents a substantial underestimation of the global costs incurred. We offer recommendations for improving estimates of costs to fill these knowledge gaps including: systematic distinction between native and invasive rodents’ impacts; monetizing indirect impacts on human health; and greater integrative and concerted research effort between scientists and stakeholders. Finally, we discuss why and how this approach will stimulate and provide support for proactive and sustainable management strategies in the context of alien rodent invasions, for which biosecurity measures should be amplified globally. creator: Christophe Diagne creator: Liliana Ballesteros-Mejia creator: Ross N. Cuthbert creator: Thomas W. Bodey creator: Jean Fantle-Lepczyk creator: Elena Angulo creator: Alok Bang creator: Gauthier Dobigny creator: Franck Courchamp uri: https://doi.org/10.7717/peerj.14935 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Diagne et al. title: N6-methyladenosine reader YTHDF1 regulates the proliferation and migration of airway smooth muscle cells through m6A/cyclin D1 in asthma link: https://peerj.com/articles/14951 last-modified: 2023-03-24 description: Asthma is a chronic inflammatory respiratory disease, which is involved in multiple pathologic molecular mechanisms and presents a huge challenge to clinic nursing. Emerging evidence suggests that N6-methyladenosine (m6A) plays critical roles in respiratory system disease. Thus, present work tried to investigate the functions of m6A reader YTHDF 1 in asthma. The results indicated that YTHDF1 significantly upregulated in platelet-derived growth factor (PDGF) induced airway smooth muscle cells (ASMCs). Functionally, overexpression of YTHDF1 promoted the proliferation and migration of ASMCs, while YTHDF1 knockdown repressed the proliferation and migration. Mechanistically, there was a m6A modification site on cyclin D1 RNA (CCND1 genome) and YTHDF1 combined with cyclin D1 mRNA, thereby enhancing its mRNA stability via m6A-dependent manner. Collectively, these findings reveal a novel axis of YTHDF1/m6A/cyclin D1 in asthma’s airway remodeling, which may provide novel therapeutic strategy for asthma. creator: Juan Wang creator: Lei Wang creator: Xingfeng Tian creator: Lingping Luo uri: https://doi.org/10.7717/peerj.14951 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Wang et al. title: Systematic review of marine environmental DNA metabarcoding studies: toward best practices for data usability and accessibility link: https://peerj.com/articles/14993 last-modified: 2023-03-24 description: The emerging field of environmental DNA (eDNA) research lacks universal guidelines for ensuring data produced are FAIR–findable, accessible, interoperable, and reusable–despite growing awareness of the importance of such practices. In order to better understand these data usability challenges, we systematically reviewed 60 peer reviewed articles conducting a specific subset of eDNA research: metabarcoding studies in marine environments. For each article, we characterized approximately 90 features across several categories: general article attributes and topics, methodological choices, types of metadata included, and availability and storage of sequence data. Analyzing these characteristics, we identified several barriers to data accessibility, including a lack of common context and vocabulary across the articles, missing metadata, supplementary information limitations, and a concentration of both sample collection and analysis in the United States. While some of these barriers require significant effort to address, we also found many instances where small choices made by authors and journals could have an outsized influence on the discoverability and reusability of data. Promisingly, articles also showed consistency and creativity in data storage choices as well as a strong trend toward open access publishing. Our analysis underscores the need to think critically about data accessibility and usability as marine eDNA metabarcoding studies, and eDNA projects more broadly, continue to proliferate. creator: Meghan M. Shea creator: Jacob Kuppermann creator: Megan P. Rogers creator: Dustin Summer Smith creator: Paul Edwards creator: Alexandria B. Boehm uri: https://doi.org/10.7717/peerj.14993 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Shea et al. title: An epidemiological study of the predictors of multidrug resistance and methicillin resistance among Staphylococcus spp. isolated from canine specimens submitted to a diagnostic laboratory in Tennessee, USA link: https://peerj.com/articles/15012 last-modified: 2023-03-24 description: BackgroundUnderstanding drivers of multidrug resistance (MDR) and methicillin resistance, which have increased among canine staphylococcal isolates, is essential for guiding antimicrobial use practices. Therefore, the objective of this study was to identify predictors of MDR and methicillin resistance among Staphylococcus spp. commonly isolated from canine clinical specimens.MethodsThis retrospective study used records of canine specimens submitted to the University of Tennessee College of Veterinary Medicine Clinical Bacteriology Laboratory for bacterial culture and antimicrobial susceptibility testing between 2006 and 2017. Records from 7,805 specimens positive for the following Staphylococcus species were included for analysis: Staphylococcus pseudintermedius, Staphylococcus aureus, Staphylococcus coagulans (formerly Staphylococcus schleiferi subspecies coagulans), and Staphylococcus schleiferi (formerly S. schleiferi subsp. schleiferi). Generalized linear regression models were fit using generalized estimating equations (GEE) to identify predictors of MDR (defined as resistance to three or more antimicrobial classes) and methicillin resistance among these isolates.ResultsMultidrug resistance (42.1%) and methicillin resistance (31.8%) were relatively common. Isolates from skeletal (joint and bone) specimens had the highest levels of MDR (51.3%) and methicillin resistance (43.6%), followed by cutaneous specimens (45.8% multidrug-resistant, 37.1% methicillin resistant). Staphylococcus species, specimen site, and clinical setting were significant (p < 0.01) predictors of both outcomes. Compared to S. pseudintermedius, S. schleiferi had higher odds of methicillin resistance, while S. coagulans and S. schleiferi had lower odds of MDR. The odds of both MDR and methicillin resistance for isolates from hospital patient specimens were significantly higher than those from referral patients for urine/bladder and otic specimens. Odds of MDR among isolates from skeletal specimens of hospital patients were also higher than those of referral patients.ConclusionsStaphylococcus isolates in this study had substantial levels of MDR and methicillin resistance. Differences in the odds of these outcomes between referral and hospital patient isolates did not persist for all specimen sites, which may reflect differences in diagnostic testing and antimicrobial use practices with respect to body site or system. Judicious antimicrobial use, informed by culture and susceptibility testing, is important to limit treatment failures and curb selection pressure. creator: Jennifer Lord creator: Nick Millis creator: Rebekah Duckett Jones creator: Brian Johnson creator: Stephen A. Kania creator: Agricola Odoi uri: https://doi.org/10.7717/peerj.15012 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Lord et al. title: A new model predicts hepatocellular carcinoma in patients with HBV-related decompensated liver cirrhosis and long-term antiviral therapy: a prospective study link: https://peerj.com/articles/15014 last-modified: 2023-03-24 description: BackgroundWe aimed to evaluate the prediction values of non-invasive models for hepatocellular carcinoma (HCC) development in patients with HBV-related liver cirrhosis (LC) and long-term NA treatment.MethodsPatients with compensated or decompensated cirrhosis (DC), who achieved long-term virological response, were enrolled. DC and its stages were defined by the complications including ascites, encephalopathy, variceal bleeding, or renal failure. Prediction accuracy of several risk scores, including ALBI, CAMD, PAGE-B, mPAGE-B and aMAP, was compared.ResultsThe median follow-up duration was 37 (28–66) months. Among the 229 patients, 9 (9.57%) patients in the compensated LC group and 39 (28.89%) patients in the DC group developed HCC. The incidence of HCC was higher in the DC group ($\cal X$X2 = 12.478, P < 0.01). The AUROC of ALBI, aMAP, CAMD, PAGE-B and mPAGE-B scores were 0.512, 0.667, 0.638, 0.663, 0.679, respectively. There was no significant difference in AUROC between CAMD, aMAP, PAGE-B and mPAGE-B (all P > 0.05). Univariable analysis showed that age, DC status and platelet were associated with HCC development, and multivariable analysis showed that age and DC status (both P < 0.01) were independent risk factors for HCC development, then Model (Age_DC) was developed and its AUROC was 0.718. Another model, Model (Age_DC_PLT_TBil) consisting of age, DC stage, PLT, TBil was also developed, and its AUROC was larger than that of Model (Age_DC) (0.760 vs. 0.718). Moreover, AUROC of Model (Age_DC_PLT_TBil) was larger than the other five models (all P < 0.05). With an optimal cut-off value of 0.236, Model (Age_DC_PLT_TBil) achieved 70.83% sensitivity, 76.24% specificity.ConclusionThere is a lack of non-invasive risk scores for HCC development in HBV-related DC, and a new model consisting of age, DC stage, PLT, TBil may be an alternative. creator: Hao-dan Mao creator: Shu-qin Zheng creator: Su-hua Yang creator: Ze-yu Huang creator: Yuan Xue creator: Min Zhou uri: https://doi.org/10.7717/peerj.15014 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Mao et al. title: Experience and coping strategies of bowel dysfunction in postoperative patients with rectal cancer: a systematic review of qualitative evidence link: https://peerj.com/articles/15037 last-modified: 2023-03-24 description: AimDue to the changes of bowel physiological structure and functional disorders after rectal cancer surgery, patients will face many bowel dysfunction for a long time, which will greatly affect their quality of life. The purpose of this review is to integrate the qualitative research on the experience of bowel dysfunction and coping strategies in postoperative patients with rectal cancer.MethodsSystematic retrieval of PubMed, EMbase, Cochrane Library, CINAHL, Web of Sciences, PsycINFO, Willey and other databases was carried out by using the method of subject words and keywords. The Critical Appraisal Skill Programme (CASP) Qualitative Studies Checklist was used for Qualitative assessment. The findings were extracted from the included study and synthesized into the final themes, which was evaluated strictly in accordance with the ConQual process.ResultsNine studies involving 345 participants were included and two main themes were extracted: “Experience a series of changes caused by bowel dysfunction” and “nmet needs and coping strategies facing bowel dysfunction”. The changes of rectal cancer patients who experience bowel dysfunction after operation mainly include three parts: bowel dysfunction is more than just a bowel reaction, which covers the bowel symptoms themselves and the subsequent body-related symptoms. The interruption of a normal life, mainly reflected in personal, family, and social life. Complex psychological reactions to bowel dysfunction, psychological changes have a dual nature, showing a positive and negative intertwined. There are two main aspects of unmet needs and coping strategies: the demand is mainly manifested in the need for information and support from medical professionals, while the coping strategy mainly includes diet, activity and drug management.ConclusionRectal cancer patient often experience persistent bowel dysfunction after operation, which has a certain physical and mental effects. A series of new needs of postoperative patients are often not fully met, and patients often rely on their own empirical attempts to seek balance, less can get professional support. Future studies need to focus on how to provide continuous information support for postoperative rectal cancer patients, especially professional care from health care staff. creator: Zhang Yanting creator: Dandan Xv creator: Wenjia Long creator: Jingyi Wang creator: Chen Tang creator: Maohui Feng creator: Xuanfei Li creator: Bei Wang creator: Jun Zhong uri: https://doi.org/10.7717/peerj.15037 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Yanting et al. title: Exploring Korean adolescent stress on social media: a semantic network analysis link: https://peerj.com/articles/15076 last-modified: 2023-03-24 description: BackgroundConsidering that adolescents spend considerable time on the Internet and social media and experience high levels of stress, it is difficult to find a study that investigates adolescent stress through a big data-based network analysis of social media. Hence, this study was designed to provide basic data to establish desirable stress coping strategies for adolescents based on a big data-based network analysis of social media for Korean adolescent stress. The purpose of this study was to (1) identify social media words that express stress in adolescents and (2) investigate the associations between those words and their types.MethodsTo analyse adolescent stress, we used social media data collected from online news and blog websites and performed semantic network analysis to understand the relationships among keywords extracted in the collected data.ResultsThe top five words used by Korean adolescents were counselling, school, suicide, depression, and activity in online news, and diet, exercise, eat, health, and obesity in blogs. As the top keywords of the blog are mainly related to diet and obesity, it reflects adolescents’ high degree of interest in their bodies; the body is also a primary source of adolescent stress. In addition, blogs contained more content about the causes and symptoms of stress than online news, which focused more on stress resolution and coping. This highlights the trend that social blogging is a new channel for sharing personal information.ConclusionsThe results of this study are valuable as they were derived through a social big data analysis of data obtained from online news and blogs, providing a wide range of implications related to adolescent stress. Hence this study can contribute basic data for the stress management of adolescents and their mental health management in the future. creator: JongHwi Song creator: JunRyul Yang creator: SooYeun Yoo creator: KyungIn Cheon creator: SangKyun Yun creator: YunHee Shin uri: https://doi.org/10.7717/peerj.15076 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Song et al. title: Baseline gene expression in subcutaneous adipose tissue predicts diet-induced weight loss in individuals with obesity link: https://peerj.com/articles/15100 last-modified: 2023-03-24 description: BackgroundWeight loss effectively reduces cardiometabolic health risks among people with overweight and obesity, but inter-individual variability in weight loss maintenance is large. Here we studied whether baseline gene expression in subcutaneous adipose tissue predicts diet-induced weight loss success.MethodsWithin the 8-month multicenter dietary intervention study DiOGenes, we classified a low weight-losers (low-WL) group and a high-WL group based on median weight loss percentage (9.9%) from 281 individuals. Using RNA sequencing, we identified the significantly differentially expressed genes between high-WL and low-WL at baseline and their enriched pathways. We used this information together with support vector machines with linear kernel to build classifier models that predict the weight loss classes.ResultsPrediction models based on a selection of genes that are associated with the discovered pathways ‘lipid metabolism’ (max AUC = 0.74, 95% CI [0.62–0.86]) and ‘response to virus’ (max AUC = 0.72, 95% CI [0.61–0.83]) predicted the weight-loss classes high-WL/low-WL significantly better than models based on randomly selected genes (P < 0.01). The performance of the models based on ‘response to virus’ genes is highly dependent on those genes that are also associated with lipid metabolism. Incorporation of baseline clinical factors into these models did not noticeably enhance the model performance in most of the runs. This study demonstrates that baseline adipose tissue gene expression data, together with supervised machine learning, facilitates the characterization of the determinants of successful weight loss. creator: Ali Oghabian creator: Birgitta W. van der Kolk creator: Pekka Marttinen creator: Armand Valsesia creator: Dominique Langin creator: W. H. Saris creator: Arne Astrup creator: Ellen E. Blaak creator: Kirsi H. Pietiläinen uri: https://doi.org/10.7717/peerj.15100 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Oghabian et al. title: Influence of mental energy on volleyball competition performance: a field test link: https://peerj.com/articles/15109 last-modified: 2023-03-24 description: Athletic mental energy is a newly emerging research topic in sport science. However, whether it can predict objective performance in competition remains unexplored. Thus, the purpose of this study was to examine the predictability of mental energy on volleyball competition performance. We recruited 81 male volleyball players (Mage = 21.11 years ± SD = 1.81) who participated in the last 16 remaining teams in a college volleyball tournament. We assessed participants’ mental energy the night before the competition and collected their competition performance over the next 3 days. We used six indices of the Volleyball Information System (VIS) developed by the International Volleyball Federation (FIVB) to examine its associations with mental energy. All six factors of mental energy –motivation, tirelessness, calm, vigor, confidence, and concentration correlated with volleyball competition performance. Further, a hierarchical regression found mental energy predicted volleyball receivers’ performance (R2 = .23). The findings advance our knowledge of mental energy and objective performance in competition. We suggest that future studies may examine the effects of mental energy on different sports with different performance indices. creator: Shiow-Fang Shieh creator: Frank J.H. Lu creator: Diane L. Gill creator: Chih-Hsuan Yu creator: Shu-Ping Tseng creator: Meisam Savardelavar uri: https://doi.org/10.7717/peerj.15109 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Shieh et al. title: Ligamentum teres reconstruction using autogenous semitendinosus tendon with toggle technique in rabbits link: https://peerj.com/articles/14777 last-modified: 2023-03-23 description: BackgroundLigamentum teres (LT) has traditionally been considered a vestigial or redundant structure in humans; however, based on new studies and the evolution of hip arthroscopy, the LT injury has been viewed as a source of hip pain. Therefore, LT reconstruction can be beneficial in some cases. Rabbits have been frequently used as a model for cranial cruciate ligament reconstruction but few studies are available for ligamentum teres reconstruction.ObjectiveTo evaluate the semitendinosus tendon to replace ligamentum teres with the toggle technique, using rabbits as an experimental model.MethodsTwenty-six female Norfolk rabbits with approximately 3 months of age were divided into two equal groups after excision of ligamentum teres (LT) from the right hip joint: G1—no reconstruction of LT and capsulorrhaphy; G2—double—bundle reconstruction of the LT using semitendinosus tendon autograft. In both groups, the LT was removed from the right hip joint. In G2 the autograft was harvested from the left hind limb of the same rabbit. The rabbits were evaluated clinically at different time intervals; before surgery (M1), 48 h (M2), 15 days (M3), 30 days (M4) and 90 days (M5) after surgery.ResultsThe rabbits supported their limbs on the ground in both the groups. As complications of the procedure, four hip joints showed subluxations in the radiographic evaluation of G1; three at M4 and one at M5. In G2; two luxations of hip joints at M3 and one subluxation at M4 were seen. On ultrasound, irregular articular surface was seen in 30.8% of the rabbits that had subluxation of hip joints. Gross evaluation identified tendon graft integrity in 76.92% of the rabbits. Histological analysis revealed graft adhesion to the bone in the early phase comprised of sharpey-like collagen fibers.ConclusionThe double-bundle reconstruction of the LT using autologous semitendinosus tendon associated with the toggle rod shows an early phase of tendon graft ligamentization at 90 days post-operatively in young rabbits, but biomechanical bias suffered by the tendon during gait must be considered. creator: Rebeca Bastos Abibe creator: Sheila Canevese Rahal creator: Luciane dos Reis Mesquita creator: Danuta Doiche creator: Jeana Pereira da Silva creator: Maria Jaqueline Mamprim creator: Renata Haddad Pinho creator: Alexandre Battazza creator: Carlos Eduardo Fonseca Alves creator: W. Brian Saunders uri: https://doi.org/10.7717/peerj.14777 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Abibe et al. title: Intersexual and body size-related variation in chemical constituents from feces and cloacal products involved in intraspecific communication of a fossorial amphisbaenian link: https://peerj.com/articles/15002 last-modified: 2023-03-23 description: BackgroundMany animals rely on chemical cues for intraspecific communication. This is especially important in fossorial animals because visual restrictions of the underground environment limit the opportunities for visual communication. Previous experiments showed the ability of the amphisbaenian Trogonophis wiegmanni to discriminate between several categories of conspecifics based on chemical cues alone. However, in contrast with many other reptile species, T. wiegmanni does not have external secretory glands, but uses uncharacterized secretions from the cloaca in intraspecific chemosensory communication.MethodsUsing gas chromatography-mass spectrometry (GC-MS), we analyzed the lipophilic compounds from feces and cloacal products freshly extracted from the cloaca of male and female T. wiegmanni. We identified and estimated relative proportions of the compounds found, and tested for intersexual and body-size related differences.ResultsWe found a total of 103 compounds, being some steroids (mainly cholesterol and cholestanol), some alkanes and squalene the most abundant and frequent. Further, we found intersexual differences, with males, especially larger ones, having higher proportions of several alkanes between C13 and C24 and of squalene than females, which had higher proportions of several steroids and also of nonacosane and methylnonacosane than males. We compared these findings with secretions of other animals and discuss the potential role of these compounds and their variations in intraspecific communication of amphisbaenians. creator: José Martín creator: Gonzalo Rodríguez-Ruiz creator: José Javier Cuervo creator: Pilar López uri: https://doi.org/10.7717/peerj.15002 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Martín et al. title: ceRNA network construction and identification of hub genes as novel therapeutic targets for age-related cataracts using bioinformatics link: https://peerj.com/articles/15054 last-modified: 2023-03-23 description: BackgroundThe aim of this study is to investigate the genetic and epigenetic mechanisms involved in the pathogenesis of age-related cataract (ARC).MethodsWe obtained the transcriptome datafile of th ree ARC samples and three healthy, age-matched samples and used differential expression analyses to identify the differentially expressed genes (DEGs). The differential lncRNA-associated competing endogenous (ceRNA) network, and the protein-protein network (PPI) were constructed using Cytoscape and STRING. Cluster analyses were performed to identify the underlying molecular mechanisms of the hub genes affecting ARC progression. To verify the immune status of the ARC patients, immune-associated analyses were also conducted.ResultsThe PPI network identified the FOXO1 gene as the hub gene with the highest score, as calculated by the Maximal Clique Centrality (MCC) algorithm. The ceRNA network identified lncRNAs H19, XIST, TTTY14, and MEG3 and hub genes FOXO1, NOTCH3, CDK6, SPRY2, and CA2 as playing key roles in regulating the pathogenesis of ARC. Additionally, the identified hub genes showed no significant correlation with an immune response but were highly correlated with cell metabolism, including cysteine, methionine, and galactose.DiscussionThe findings of this study may provide clues toward ARC pathogenic mechanisms and may be of significance for future therapeutic research. creator: Yingying Hong creator: Jiawen Wu creator: Yang Sun creator: Shenghai Zhang creator: Yi Lu creator: Yinghong Ji uri: https://doi.org/10.7717/peerj.15054 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Hong et al. title: Common eating habit patterns are associated with a high maximum occlusal force and pre-eating cardiac vagal tone link: https://peerj.com/articles/15091 last-modified: 2023-03-23 description: BackgroundMasticatory function is associated with nervous function, including autonomic nervous function, and both functions are influenced by human habits. In a previous preliminary study of 53 young women, we found that eating habit patterns were associated with occlusal force as an indicator of masticatory function. Therefore, we hypothesized that relationships exist between occlusal force, the autonomic nervous system, and eating habit patterns.MethodsTo test our hypothesis, we analyzed the relationship between heart rate variability measured before and after lunch in 53 young women, and measured and surveyed maximum occlusal force and eating habit patterns, respectively, in these participants.ResultsHigh occlusal force was associated with an increased high-frequency (HF) component (vagal tone index) of the heart rate variability index immediately before lunch (standardized regression coefficient (β) = 0.417, P = 0.002). Each of the eating habit items surveyed in a questionnaire showed a similar tendency for the HF component immediately before lunch and maximum occlusal force; in particular, “Habit of eating breakfast” and “Number of meals per day” were significantly associated with both variables. Additionally, total eating habit scores related to higher maximum occlusal force were associated with an increase in the HF component immediately before lunch (β = 0.514, P < 0.001). The maximum occlusal force and the pre-eating HF component values were stratified by total eating habit scores (into low, medium, high categories), and the high scores were significantly higher than the medium or low scores.ConclusionsOcclusal force and the pre-eating cardiac vagal response of individuals were characterized by their common eating habit patterns, indicating that eating habits may be simultaneously associated with the development of masticatory function, nervous system development, and cardiovascular rhythm. Although further research is needed to investigate these relationships in detail, our findings provide insights that will inform the study of physical functions, neurodevelopment, habitual behaviors, and health in humans. creator: Masahiro Okada creator: Kosuke Okada creator: Masayuki Kakehashi uri: https://doi.org/10.7717/peerj.15091 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Okada et al. title: Hydrogels improved parsley (Petroselinium crispum(Mill.) Nyman) growth and development under water deficit stress link: https://peerj.com/articles/15105 last-modified: 2023-03-23 description: Water scarcity is one of the most pressing problems facing countries in the semi-arid and arid regions of the world. Data predicts that by 2030, global water consumption will increase by 50%, leading to severe water shortages. Today, agricultural production consumes more than 70% of fresh water in many parts of the world, increasing the pressure on water scarcity. For these reasons, agricultural production models and approaches should be developed to reduce water consumption. One developed approach is the use of hydrogel to reduce water consumption and have a positive effect on plant growth. This study investigated the use of hydrogels as chemical components that can be used in water shortage conditions and against the expected water scarcity. Parsley was used as the model organism. The method used was as follows: two different water treatments (50% and 100%) and four different hydrogel concentrations (0%, 50%, 75%, and 100%) were applied, and root width and length, leaf width and length, main stem length, and the number of tillers were measured. According to the results, while no improvement was observed in the plants with 100% hydrogel concentration, the best results were obtained from 50% hydrogel application. The results obtained from 75% hydrogel application were found to be higher than those of 100% hydrogel but lower than 0% hydrogel application. With 50% hydrogel (water-restricted), all plant growth parameters were higher compared to the plants with 100% (full irrigation) water application. It was determined that the average value of the I1 (50%) irrigation was the highest (3.6), and the average value of the I2 (100%) irrigation (2.4) was the lowest. It was determined that the highest average value (6.2) in all measured traits was the average value of the H1 (50%) application, and the lowest average value (0.0) was in the H3 hydrogel applications (100%). In conclusion, this study suggested that hydrogel application is beneficial on a large scale, can optimize water resource management for higher yields in agriculture, and has a positive effect on agricultural yield under water deficit stress. creator: M. Zeki Karipçin uri: https://doi.org/10.7717/peerj.15105 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Karipçin title: Environmental and individual determinants of burrow-site microhabitat selection, occupancy, and fidelity in eastern chipmunks living in a pulsed-resource ecosystem link: https://peerj.com/articles/15110 last-modified: 2023-03-23 description: BackgroundHabitat selection has major consequences on individual fitness, particularly selection for breeding sites such as nests or burrows. Theory predicts that animals will first use optimal habitats or rearrange their distribution by moving to higher-quality habitats whenever possible, for instance when another resident disperses or dies, or when environmental changes occur. External constraints, such as predation risk or resource abundance, and interindividual differences in age, sex and body condition can lead to variation in animals’ perception of habitat quality. Following habitat use by individuals over their lifetime is thus essential to understand the causes of variation in habitat selection within a population.MethodsWe used burrow occupancy data collected over eight years to assess burrow-site selection in a population of wild eastern chipmunks (Tamias striatus) relying on pulsed resources. We first compared characteristics of burrow microhabitats with those of equivalent unused plots. We then investigated the factors influencing the frequency of burrow occupation over time, and the individual and environmental causes of annual burrow fidelity decisions.ResultsOur results indicate that chipmunks select microhabitats with a greater number of woody debris and greater slopes. Microhabitats of burrows with higher occupancy rates had a lower shrub stratum, were less horizontally opened and their occupants’ sex-ratio was skewed towards males. Burrow fidelity was higher in non-mast years and positively related to the occupant’s age, microhabitat canopy cover and density of large red maples.ConclusionThe quality of a burrow microhabitat appears to be determined in part by characteristics that favour predation avoidance, but consideration of occupancy and fidelity patterns over several years also highlighted the importance of including individual and contextual factors in habitat selection studies. creator: Camille Gaudreau-Rousseau creator: Patrick Bergeron creator: Denis Réale creator: Dany Garant uri: https://doi.org/10.7717/peerj.15110 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Gaudreau-Rousseau et al. title: Vibrations on mastoid process alter the gait characteristics during walking on different inclines link: https://peerj.com/articles/15111 last-modified: 2023-03-23 description: BackgroundEighty-eight percent of the persons with bilateral vestibular dysfunction have reported at least one fall within the past 5 years. The apparent alternations due to the bilateral vestibular dysfunctions (BVD) are the gait characteristics, such as slower walking speed, prolonged stance phase, and shorter step length. Unexpectedly, due to the prevalence of this BVD being relatively low, attention is not obtained as same as in other vestibular disorders. Moreover, how does walking on different inclines, part of daily activities, alter the gait characteristics under the unreliable bilateral vestibular systems? Previous studies used vibration-based stimulations (VS) as a perturbation to understand the postural control during walking while the bilateral vestibular systems were perturbed. Therefore, this study attempted to extend the knowledge to understand the alternations in spatial-temporal gait characteristics under perturbed bilateral vestibular systems while walking on different inclines.MethodsNineteen healthy young adults participated in this study. Eight walking conditions were randomly assigned to each participant: 0%, 3%, 6%, and 9% grade of inclines with/without VS. The preferred walking speed was used for gait analysis. The dependent variables were stance time, double support time, step length, step time, step width, foot clearance, and respective variabilities. All dependent variables were defined by two critical gait events: heel-strike and toe-off. Pre-Hoc paired comparisons with Bonferroni corrections were used to prioritize the dependent variables. A two-way repeated measure was used to investigate the effect of VS and the effect of inclines on the selected dependent variables from Pre-Hoc analysis. Post-Hoc comparisons were also corrected by the Bonferroni method.ResultsThe step length, step time, foot clearance, and foot clearance variability were selected by the Pre-Hoc analysis because the corrected paired t-test demonstrated a significant VS effect (p < 0.05) on these gait parameters at least one of four inclines. The significant interaction between the effect of VS and the effect of inclines was found in step length (p = 0.005), step time (p = 0.028), and foot clearance variability (p = 0.003). The results revealed that implementing a VS increased step length and step time when walking on 0%, 3%, and 9% of grade inclines. In particular, the foot clearance variability was found when walking on 9% of grade inclines.ConclusionThe observations in the current study suggested that VS increased the step length, step time, foot clearance, and foot clearance variability while walking on inclines. These results suggested that these gait parameters might be promising targets for future clinical investigations in patients with BVD while walking on different inclines. Importantly, the increases in spatial-temporal gait performance under bilateral VS might be an indicator of gait improvement while walking on different inclines. creator: Yuxiao Sun creator: Dongqi Zhu creator: Huiyan Song creator: Jung H. Chien uri: https://doi.org/10.7717/peerj.15111 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Sun et al. title: Integrating eDNA and citizen science observations to model distribution of a temperate freshwater turtle near its northern range limit link: https://peerj.com/articles/15120 last-modified: 2023-03-23 description: BackgroundTo determine species distributions and the factors underlying them, reliable occurrence data are crucial. Assembling such data can be challenging for species with cryptic life histories or that occur at low densities.MethodsWe developed species-specific eDNA protocols, from sampling through data interpretation, to detect the common musk turtle (Sternotherus odoratus) and tested whether eDNA occurrences change our understanding of the species distribution and the factors that shape its northern range limit. We used Species Distribution Models (SDMs) with full parameter optimization on citizen science observations of S. odoratus in Southern Ontario alone and together with eDNA occurrences.ResultsOur eDNA protocol was robust and sensitive. SDMs built from traditional observations and those supplemented with eDNA detections were comparable in prediction accuracy. However, models with eDNA detections suggested that the distribution of S. odoratus in Southern Ontario is underestimated, especially near its northern range limit, and that it is shaped by thermal conditions, hydrology, and elevation. Our study underscores the promise of eDNA for surveying cryptic aquatic organisms in undocumented areas, and how such insights can help us to improve our understanding of species distributions. creator: Wenxi Feng creator: Stephen C. Lougheed uri: https://doi.org/10.7717/peerj.15120 license: https://creativecommons.org/licenses/by-nc/4.0 rights: © 2023 Feng and Lougheed title: Epoch and accuracy based empirical study for cardiac MRI segmentation using deep learning technique link: https://peerj.com/articles/14939 last-modified: 2023-03-22 description: Cardiac magnetic resonance imaging (CMRI) is a non-invasive imaging technique to analyse the structure and function of the heart. It was enhanced considerably over several years to deliver functional information for diagnosing and managing cardiovascular disease. CMRI image delivers non-invasive, clear access to the heart and great vessels. The segmentation of CMRI provides quantification parameters such as myocardial viability, ejection fraction, cardiac chamber volume, and morphological details. In general, experts interpret the CMR images by delineating the images manually. The manual segmentation process is time-consuming, and it has been observed that the final observation varied with the opinion of the different experts. Convolution neural network is a new-age technology that provides impressive results compared to manual ones. In this study convolution neural network model is used for the segmentation task. The neural network parameters have been optimized to perform on the novel data set for accurate predictions. With other parameters, epochs play an essential role in training the network, as the network should not be under-fitted or over-fitted. The relationship between the hyperparameter epoch and accuracy is established in the model. The model delivers the accuracy of 0.88 in terms of the IoU coefficient. creator: Niharika Das creator: Sujoy Das uri: https://doi.org/10.7717/peerj.14939 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Das and Das title: Finding priority bacterial ribosomes for future structural and antimicrobial research based upon global RNA and protein sequence analysis link: https://peerj.com/articles/14969 last-modified: 2023-03-22 description: Ribosome-targeting antibiotics comprise over half of antibiotics used in medicine, but our fundamental knowledge of their binding sites is derived primarily from ribosome structures of non-pathogenic species. These include Thermus thermophilus, Deinococcus radiodurans and the archaean Haloarcula marismortui, as well as the commensal and sometimes pathogenic organism, Escherichia coli. Advancements in electron cryomicroscopy have allowed for the determination of more ribosome structures from pathogenic bacteria, with each study highlighting species-specific differences that had not been observed in the non-pathogenic structures. These observed differences suggest that more novel ribosome structures, particularly from pathogens, are required for a more accurate understanding of the level of diversity of the entire bacterial ribosome, with the potential of leading to innovative advancements in antibiotic research. In this study, high accuracy covariance and hidden Markov models were used to annotate ribosomal RNA and protein sequences respectively from genomic sequence, allowing us to determine the underlying ribosomal sequence diversity using phylogenetic methods. This analysis provided evidence that the current non-pathogenic ribosome structures are not sufficient representatives of some pathogenic bacteria, such as Campylobacter pylori, or of whole phyla such as Bacteroidota (Bacteroidetes). creator: Helena B. Cooper creator: Kurt L. Krause creator: Paul P. Gardner uri: https://doi.org/10.7717/peerj.14969 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Cooper et al. title: A new juvenile sauropod specimen from the Middle Jurassic Dongdaqiao Formation of East Tibet link: https://peerj.com/articles/14982 last-modified: 2023-03-22 description: Jurassic strata are widely distributed in the eastern part of Tibet Autonomous Region, and have yielded many dinosaur bones. However, none of these specimens has been studied extensively, and some remain unprepared. Here we provide a detailed description of some new sauropod material, including several cervical vertebrae and a nearly complete scapula, recovered from the Middle Jurassic of Chaya County, East Tibet. The cervical vertebrae have short centra that bear ventral midline keels, as in many non-neosauropod sauropods such as Shunosaurus. Moreover, the cervical centra display deep lateral excavations, partitioned by a septum. The scapula has proximal and distal ends that are both expanded as in mamenchisaurids and neosauropods. However, relatively small body size and lack of fusion of neurocentral sutures in the cervical vertebrae suggest that the available material is from a juvenile, and the length of the cervical centra may have increased relative to the size of the rest of the skeleton in later ontogenetic stages. Phylogenetic analysis provides limited evidence that the new Tibetan sauropod specimen belongs to Eusauropoda, being more derived than Shunosaurus, but is basal to Mamenchisauridae. The new material provides important information on the morphological transition between Shunosaurus and mamenchisaurids, and extends the known biogeographic range of early-diverging sauropods in the Middle Jurassic of East Asia. creator: Xianyin An creator: Xing Xu creator: Fenglu Han creator: Corwin Sullivan creator: Qiyu Wang creator: Yong Li creator: Dongbing Wang creator: Baodi Wang creator: Jinfeng Hu uri: https://doi.org/10.7717/peerj.14982 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 An et al. title: Sida chlorotic leaf virus: a new recombinant begomovirus found in non-cultivated plants and Cucumis sativus L link: https://peerj.com/articles/15047 last-modified: 2023-03-22 description: BackgroundBegomoviruses are circular single-stranded DNA plant viruses that cause economic losses worldwide. Weeds have been pointed out as reservoirs for many begomoviruses species, especially from members of the Sida and Malvastrum genera. These weeds have the ability to host multiple begomoviruses species simultaneously, which can lead to the emergence of new viral species that can spread to commercial crops. Additionally, begomoviruses have a natural tendency to recombine, resulting in the emergence of new variants and species.MethodsTo explore the begomoviruses biodiversity in weeds from genera Sida and Malvastrum in Colima, México, we collected symptomatic plants from these genera throughout the state. To identify BGVs infecting weeds, we performed circular DNA genomics (circomics) using the Illumina platform. Contig annotation was conducted with the BLASTn tool using the GenBank nucleotide “nr” database. We corroborated by PCR the presence of begomoviruses in weeds samples and isolated and sequenced the complete genome of a probable new species of begomovirus using the Sanger method. The demarcation process for new species determination followed the International Committee on Taxonomy of Viruses criteria. Phylogenetic and recombination analyses were implemented to infer the evolutionary relationship of the new virus.ResultsWe identified a new begomovirus species from sida and malvastrum plants that has the ability to infect Cucumis sativus L. According to our findings, the novel species Sida chlorotic leaf virus is the result of a recombination event between one member of the group known as the Squash leaf curl virus (SLCV) clade and another from the Abutilon mosaic virus (AbMV) clade. Additionally, we isolated three previously identified begomoviruses species, two of which infected commercial crops: okra (Okra yellow mosaic Mexico virus) and cucumber (Cucumber chlorotic leaf virus).ConclusionThese findings support the idea that weeds act as begomovirus reservoirs and play essential roles in begomovirus biodiversity. Therefore, controlling their populations near commercial crops must be considered in order to avoid the harmful effects of these phytopathogens and thus increase agricultural efficiency, ensuring food and nutritional security. creator: Daniel Alejandro García-Rodríguez creator: Brenda Lizet Partida-Palacios creator: Carlos Fernando Regla-Márquez creator: Sara Centeno-Leija creator: Hugo Serrano-Posada creator: Bernardo Bañuelos-Hernández creator: Yair Cárdenas-Conejo uri: https://doi.org/10.7717/peerj.15047 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Garcia-Rodriguez et al. title: Raptors bred in captivity: semen characteristics and assisted reproduction outcome in goshawk (Accipiter gentilis) link: https://peerj.com/articles/15094 last-modified: 2023-03-22 description: Three sexually mature goshawks reared in captivity and imprinted on humans to express reproductive behavior according to the cooperative method were studied for three consecutive breeding seasons to assess the quality of their sperm. The following parameters were analyzed: ejaculate volume and sperm concentration, motility, viability, and morphology. Ejaculate volume, sperm concentration and motility fluctuated along the reproductive season, revealing the greatest quality of the reproductive material at full springtime (i.e., April). Motility of the sperm collected in March strongly reduced with age, contrary to samples collected in April or May. Sperm viability was not influenced by either age or month of collection within each season. Ultrastructural investigations provided information on normal sperm morphology for the first time in this species. The morphological categories of sperm defects in fresh semen, present at low percentages, are also described. Functional analyses (perivitelline membrane assay and artificial inseminations) confirmed the good quality of the semen obtained using the cooperative method. The reported data provide the basis for further studies aimed at developing protocols to improve the outcome of artificial insemination and semen cryopreservation in the goshawk as well as other bird of prey species. creator: Anna Maria Fausto creator: Anna Rita Taddei creator: Federica Batocco creator: Maria Cristina Belardinelli creator: Marcella Carcupino creator: Achille Schiavone creator: Sergio Saia creator: Annelisse Castillo creator: Margherita Marzoni uri: https://doi.org/10.7717/peerj.15094 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Fausto et al. title: Revisiting the genetic diversity and population structure of the endangered Green Sea Turtle (Chelonia mydas) breeding populations in the Xisha (Paracel) Islands, South China Sea link: https://peerj.com/articles/15115 last-modified: 2023-03-22 description: The Green Sea Turtle (Chelonia mydas) is an umbrella species in the South China Sea, a Chinese national first-level protected wild animal, and the only sea turtle that nests in waters around China. The largest C. mydas nesting ground is distributed in the Xisha (Paracel) Islands, which plays a vital role in the survival of sea turtle populations in the region. This study reveals the genetic diversity and population structure of the breeding population of C. mydas in the Xisha (Paracel) Islands using three mitochondrial markers. A total of 15 D-loop, five Cytochrome b (Cyt b), and seven Cytochrome C Oxidase subunit I (COI) haplotypes were identified in the breeding population of C. mydas in the Xisha (Paracel) Islands. D-loop haplotypes are distributed in clades III, IV, and VIII of the C. mydas mitochondrial control region. It is the first time that one haplotype from Clade IV was found in this C. mydas population, and five new D-loop haplotypes were also identified. The haplotype and nucleotide diversity were calculated for each marker: D-loop (0.415 haplotype diversity, 0.00204 nucleotide diversity), Cyt b (0.140, 0.00038) and COI (0.308, 0.00083). The average genetic distance (p) of each molecular marker was less than 0.01. Neutral detection and nucleotide mismatch analysis suggested that the breeding population of C. mydas in the Xisha (Paracel) Islands did not experience a population expansion event in recent history. It is recommended that a sea turtle protection area be established in the Xisha (Paracel) Islands area to strengthen protection and effectively protect the uniqueness and sustainability of the breeding population of C. mydas in the South China Sea. creator: Meimei Li creator: Ting Zhang creator: Yunteng Liu creator: Yupei Li creator: Jonathan J. Fong creator: Yangfei Yu creator: Jichao Wang creator: Hai-tao Shi creator: Liu Lin uri: https://doi.org/10.7717/peerj.15115 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Li et al. title: Allometry of bud dynamic pattern and linkage between bud traits and ecological stoichiometry of Nitraria tangutorum under fertilizer addition link: https://peerj.com/articles/14934 last-modified: 2023-03-21 description: Affected by the pressure and constraints of available resources, plant growth and development, as well as plant life history strategies, usually vary with environmental conditions. Plant buds play a crucial role in the life history of woody plants. Nitraria tangutorum is a common dominant woody species in desertified areas of northern China and its growth is critical to the desert ecosystem. Revealing the allometry of N. tangutorum aboveground bud fates and the linkage between bud traits and plant nutrient contents and stoichiometric ratios can be useful in understanding plant adaptation strategy. We applied seven nitrogen and phosphorus fertilizer addition treatments to natural N. tangutorum ramets in Ulan Buh Desert in three consecutive years. We surveyed three types of aboveground buds (dormant buds, vegetative buds, and reproductive buds) in each N. tangutorum ramet, then measured the plant carbon (C), nitrogen (N), and phosphorus (P) contents and ratios during three consecutive years. We specified that reserve growth potential (RGP), vegetative growth intensity (VGI) and sexual reproduction effort (SRE) are the three indices of bud dynamic pattern. The results showed that the bud dynamic pattern of N. tangutorum ramets differed significantly among different fertilizer addition treatments and sampling years. The allometry of RGP, VGI, and SRE was obvious, showing size dependence. The allometric growth relationship fluctuated among the sampling years. The linkage between bud traits and plant stoichiometric characteristics of N. tangutorum ramets showed close correlation with plant P content, C:P and N:P ratios, no significant correlation with plant C content, N content and C:N ratio. These results contribute to an improved understanding of the adaptive strategies of woody plants growing in desert ecosystems and provide insights for adoption of effective measures to restore and conserve plant communities in arid and semi-arid regions. creator: Qinghe Li creator: Na Duan creator: Chenggong Liu creator: Huiqing Li creator: Lan Xu uri: https://doi.org/10.7717/peerj.14934 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Li et al. title: Prediction of HIV-1 protease resistance using genotypic, phenotypic, and molecular information with artificial neural networks link: https://peerj.com/articles/14987 last-modified: 2023-03-21 description: Drug resistance is a primary barrier to effective treatments of HIV/AIDS. Calculating quantitative relations between genotype and phenotype observations for each inhibitor with cell-based assays requires time and money-consuming experiments. Machine learning models are good options for tackling these problems by generalizing the available data with suitable linear or nonlinear mappings. The main aim of this study is to construct drug isolate fold (DIF) change-based artificial neural network (ANN) models for estimating the resistance potential of molecules inhibiting the HIV-1 protease (PR) enzyme. Throughout the study, seven of eight protease inhibitors (PIs) have been included in the training set and the remaining ones in the test set. We have obtained 11,803 genotype-phenotype data points for eight PIs from Stanford HIV drug resistance database. Using the leave-one-out (LVO) procedure, eight ANN models have been produced to measure the learning capacity of models from the descriptors of the inhibitors. Mean R2 value of eight ANN models for unseen inhibitors is 0.716, and the 95% confidence interval (CI) is [0.592–0.840]. Predicting the fold change resistance for hundreds of isolates allowed a robust comparison of drug pairs. These eight models have predicted the drug resistance tendencies of each inhibitor pair with the mean 2D correlation coefficient of 0.933 and 95% CI [0.930–0.938]. A classification problem has been created to predict the ordered relationship of the PIs, and the mean accuracy, sensitivity, specificity, and Matthews correlation coefficient (MCC) values are calculated as 0.954, 0.791, 0.791, and 0.688, respectively. Furthermore, we have created an external test dataset consisting of 51 unique known HIV-1 PR inhibitors and 87 genotype-phenotype relations. Our developed ANN model has accuracy and area under the curve (AUC) values of 0.749 and 0.818 to predict the ordered relationships of molecules on the same strain for the external dataset. The currently derived ANN models can accurately predict the drug resistance tendencies of PI pairs. This observation could help test new inhibitors with various isolates. creator: Huseyin Tunc creator: Berna Dogan creator: Büşra Nur Darendeli Kiraz creator: Murat Sari creator: Serdar Durdagi creator: Seyfullah Kotil uri: https://doi.org/10.7717/peerj.14987 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Tunc et al. title: New agri-environmental measures have a direct effect on wildlife and economy on conventional agricultural land link: https://peerj.com/articles/15000 last-modified: 2023-03-21 description: The objective of this article is to evaluate economic profits along with return on investment and also the impact of newly designed agri-environmental measures (AEM) on the presence of bioindicator species—European hare and roe deer—in comparison to conventionally cultivated agricultural land. The abundance of European hare was, on average, 4.5–6.7 times higher on AEM compared to the standard agricultural regime and 3.5–6.4 times higher in the case of roe deer in 2020 and 2021. From an economic point of view, the highest incomes were found for extensive orchard alleys and standard conventional crops—wheat and rapeseed rotation. The cash flow from extensive orchard was 4.3 times larger and wheat and rapeseed were 3.5 times larger than from the clover grass mixture. Moreover, the lowest value of operational expenses was found in the case of extensive orchard alleys. The payback period ranged from 16.02 years (wheat and rapeseed rotation) to 53.6 years (clover grass mixture). It is crucial not to assess the economic parameters separately but optimize them with sustainable wildlife management and other benefits that provide ecological and efficient directions of AEM for future generations. However, the performed economic analysis highlights the significantly lower incomes of most components of AEM. We see a principal issue of AEMs usage in the lack of strong incentives for farmers to maximize conservation outcomes. Therefore, the AEMs are often placed in locations with presumed low agriculture profit, which is often related to insignificant conservation effects. Thus, the incomparable AEMs profitability compared to conventional agriculture has to be reflected by the agricultural policy at the European Union level and subsidy policy of particular member states. creator: Petr Marada creator: Jan Cukor creator: Michal Kuběnka creator: Rostislav Linda creator: Zdeněk Vacek creator: Stanislav Vacek uri: https://doi.org/10.7717/peerj.15000 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Marada et al. title: Depression and insomnia among healthcare professionals during COVID-19 pandemic in Ethiopia: a systematic review and meta-analysis link: https://peerj.com/articles/15039 last-modified: 2023-03-21 description: IntroductionHealthcare professionals play a great role in the struggle against COVID-19. They are highly susceptible to COVID-19 due to their responsibilities. This susceptibility directly affects their mental health status. Comprehensive evidence on prevalence of depression and insomnia during this pandemic is vital. Thus, this study aims to provide the pooled prevalence of depression and insomnia, and their associated factors during the COVID-19 pandemic.Materials and methodsThis systematic review and meta-analysis follow the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) guidelines. Studies were searched from PubMed, Cochrane Library, CrossRef, African Journals Online and Google Scholar databases from the occurence of the pandemic to June 2022. Study selection, data extraction and methodological quality assessment were done by two authors independently. The I2 statistics was used for testing heterogeneity. A random effect model was used. Stata version 16.0 was used for statistical analysis.ResultsEight studies were incorporated for this systematic review and meta-analysis. From seven studies the pooled prevalence of depression was 40% (95% CI [0.23–0.57]; I2 = 99.00%; P = 0.00). From three studies the pooled prevalence of insomnia was 35% (95% CI [0.13–0.58]; I2 = 98.20; P = 0.00). Associated factors of depression on healthcare workers (HCWs) were being female pooled AOR: 2.09; 95% CI [1.41–2.76], been married (pooled AOR = 2.95; 95% CI [1.83–4.07]). Due to limited studies available on the factors associated with insomnia, it is impossible to pool and associated factors were presented in narrative synthesis.ConclusionCOVID-19 is highly associated with the prevalence of depression and insomnia among healthcare professionals in Ethiopia. The pooled prevalence of depression and insomnia were significantly higher among healthcare professionals. Appropriate psychological counseling package should be realized for healthcare workers (HCWs) in order to recover the general mental health problems. Trial registration. This review was registered PROSPERO with registration number CRD42022314865. creator: Aragaw Asfaw Hasen creator: Abubeker Alebachew Seid creator: Ahmed Adem Mohammed uri: https://doi.org/10.7717/peerj.15039 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Hasen et al. title: Identifying effects of genetic obesity exposure on leukocyte telomere length using Mendelian randomization link: https://peerj.com/articles/15085 last-modified: 2023-03-21 description: BackgroundObservational studies have shown that obesity is closely associated with leukocyte telomere length (LTL). However, the causal relationship between obesity and LTL remains unclear. This study investigated the causal relationship between obesity and LTL through the Mendelian randomization approach.Materials and MethodsThe genome-wide association study (GWAS) summary data of several studies on obesity-related traits with a sample size of more than 600,000 individuals were extracted from the UK Biobank cohort. The summary-level data of LTL-related GWAS (45 6,717 individuals) was obtained from the IEU Open GWAS database. An inverse-variance-weighted (IVW) algorithm was utilized as the primary MR analysis method. Sensitivity analyses were conducted via MR-Egger regression, IVW regression, leave-one-out test, MR-pleiotropy residual sum, and outlier methods.ResultsHigh body mass index was correlated with a short LTL, and the odds ratio (OR) was 0.957 (95% confidence interval [CI] 0.942–0.973, p = 1.17E−07). The six body fat indexes (whole body fat mass, right leg fat mass, left leg fat mass, right arm fat mass, left arm fat mass, and trunk fat mass) were consistently inversely associated with LTL. Multiple statistical sensitive analysis approaches showed that the adverse effect of obesity on LTL was steady and dependable.ConclusionThe current study provided robust evidence supporting the causal assumption that genetically caused obesity is negatively associated with LTL. The findings may facilitate the formulation of persistent strategies for maintaining LTL. creator: Bangbei Wan creator: Ning Ma creator: Cai Lv uri: https://doi.org/10.7717/peerj.15085 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Wan et al. title: Evaluating the causal relationship between five modifiable factors and the risk of spinal stenosis: a multivariable Mendelian randomization analysis link: https://peerj.com/articles/15087 last-modified: 2023-03-21 description: BackgroundSpinal stenosis is a neurological disorder related to the compression of the spinal cord or nerve roots, and its incidence increases yearly. We aimed to use Mendelian randomization (MR) to investigate the causal relationship between several modifiable risk factors and the risk of spinal stenosis.MethodsWe obtained genome-wide association study summary data of large-sample projects (more than 100,000 individuals) from public databases. The data were associated with traits, including years of schooling (educational attainment) from the IEU OpenGWAS Project, smoking behavior (never vs. initiation) from the IEU OpenGWAS Project, body mass index (BMI) from the UK Biobank, length of mobile phone use from the UK Biobank, time spent watching television (TV) from the UK Biobank, and spinal stenosis from FinnGen biobank. Spinal stenosis was used as the outcome, whereas the other four traits were used as exposures. Inverse variance weighted (IVW) regressions were used as a primary to estimate the causal-effect size. Several sensitive analyses (including consistency, heterogenicity, and pleiotropy analyses) were conducted to test the stability and reliability of causal estimates.ResultsUnivariable MR analyses showed that genetically predicted higher educational attainment (IVW; odds ratio (OR) = 0.606; 95% confidence interval (CI): 0.507–0.724; P = 3.37 × 10−8) and never smoking (IVW; OR = 1.388; 95% CI [1.135–1.697]; P = 0.001) were negatively correlated with the risk of spinal stenosis. Meanwhile, a higher BMI (IVW; OR = 1.569; 95% CI [1.403–1.754]; P = 2.35 × 10−8), longer time spent using a mobile phone (IVW; OR = 1.895; 95% CI [1.306–2.750]; P = 0.001), and watching TV (IVW; OR = 1.776; 95% CI [1.245–2.532]; P = 0.002) were positively associated with the risk of spinal stenosis. Multivariable MR analysis indicated that educational attainment (IVW; OR = 0.670; 95% CI [0.465–0.967]; P = 0.032) and BMI (IVW; OR = 1.365; 95% CI [1.179–1.580]; P = 3.12 × 10−5) were independently causally related to the risk of spinal stenosis.ConclusionOur findings supported the potential causal associations of the five factors (educational attainment, smoking behavior, BMI, length of mobile phone use, and watching TV) with the risk for spinal stenosis. While replication studies are essential, these findings may provide a new perspective on prevention and intervention strategies directed toward spinal stenosis. creator: Bangbei Wan creator: Ning Ma creator: Weiying Lu uri: https://doi.org/10.7717/peerj.15087 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Wan et al. title: Ideal family size decision and its associated factors among women of reproductive age: community survey in southern Ethiopia link: https://peerj.com/articles/15103 last-modified: 2023-03-21 description: BackgroundUnderstanding fertility preferences is important for population studies and planning programs. The ideal family size, which is the number of children wanted in one’s lifetime, is one of variables used to measure fertility preferences. However, there was limited information on ideal family size decision in Southern Ethiopia. Therefore, this study was designed to assess ideal family size decision and its associated factors among women of reproductive age in Southern Ethiopia.MethodsA community based cross-sectional study was conducted in 2015 in Southern Ethiopia. Multi-stage stratified cluster sampling was used to select 3,205 study subjects. Data on socio-demographic characteristics, reproductive history, deciding ideal number of children, knowledge and utilization of contraceptives were considered. Descriptive statistics and binary logistic regression were done to describe and assess factors associated with deciding ideal family size. The association between variables was presented using odds ratios with 95% confidence intervals.ResultsWe included 3,205 women of reproductive age from which 37.5% respondents could not read and write and 56.5% were housewives. About half of the respondents, 47.1%, did not decide ideal numeric family size or failed to report numeric preferences while 21.6% desire to have five or more children. The mean ideal number of children preferred was 4.5 ± 1.62. Educational status (P < 0.001), overall knowledge about contraceptives (P < 0.001), current contraceptive use (P < 0.001), place of residence (P < 0.001), age (P < 0.004), marital status (P < 0.003) and number of living children (P < 0.003) were factors associated with deciding ideal family size.ConclusionOnly about half of respondents decided ideal family size from which one fifth prefer high fertility. The mean ideal number of children was comparable with that of Sub-Saharan estimate. Counselling that can empower women to decide family size should be strengthened to empower less empowered women. creator: Misganu Endriyas creator: Agegnehu Gebru creator: Amare Assefa uri: https://doi.org/10.7717/peerj.15103 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Endriyas et al. title: Metabolomic responses to the mechanical wounding of Catharanthus roseus’ upper leaves link: https://peerj.com/articles/14539 last-modified: 2023-03-20 description: PurposePlant secondary metabolites are used to treat various human diseases. However, it is difficult to produce a large number of specific metabolites, which largely limits their medicinal applications. Many methods, such as drought and nutrient application, have been used to induce the biosynthetic production of secondary metabolites. Among these secondary metabolite-inducing methods, mechanical wounding maintains the composition of secondary metabolites with little potential risk. However, the effects of mechanical stress have not been fully investigated, and thus this method remains widely unused.MethodsIn this study, we used metabolomics to investigate the metabolites produced in the upper and lower leaves of Catharanthus roseus in response to mechanical wounding.ResultsIn the upper leaves, 13 different secondary metabolites (three terpenoid indole alkaloids and 10 phenolic compounds) were screened using an orthogonal partial least squares discriminant analysis (OPLS-DA) score plot. The mechanical wounding of different plant parts affected the production of secondary metabolites. Specifically, when lower leaves were mechanically wounded, the upper leaves became a strong source of resources. Conversely, when upper leaves were injured, the upper leaves themselves became a resource sink. Changes in the source-sink relationship reflected a new balance between resource tradeoff and the upregulation or downregulation of certain metabolic pathways.ConclusionOur findings suggest that mechanical wounding to specific plant parts is a novel approach to increase the biosynthetic production of specific secondary metabolites. These results indicate the need for a reevaluation of production practices for secondary metabolites from select commercial plants. creator: Qi Chen creator: Yan Jin creator: Xiaorui Guo creator: Mingyuan Xu creator: Guanyun Wei creator: Xueyan Lu creator: Zhonghua Tang uri: https://doi.org/10.7717/peerj.14539 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Chen et al. title: The effect of node features on GCN-based brain network classification: an empirical study link: https://peerj.com/articles/14835 last-modified: 2023-03-20 description: Brain functional network (BFN) analysis has become a popular technique for identifying neurological/mental diseases. Due to the fact that BFN is a graph, a graph convolutional network (GCN) can be naturally used in the classification of BFN. Different from traditional methods that directly use the adjacency matrices of BFNs to train a classifier, GCN requires an additional input-node features. To our best knowledge, however, there is no systematic study to analyze their influence on the performance of GCN-based brain disorder classification. Therefore, in this study, we conduct an empirical study on various node feature measures, including (1) original fMRI signals, (2) one-hot encoding, (3) node statistics, (4) node correlation, and (5) their combination. Experimental results on two benchmark databases show that different node feature inputs to GCN significantly affect the brain disease classification performance, and node correlation usually contributes higher accuracy compared to original signals and manually extracted statistical features. creator: Guangyu Wang creator: Limei Zhang creator: Lishan Qiao uri: https://doi.org/10.7717/peerj.14835 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Wang et al. title: Genome-wide identification and expression analysis of diacylglycerol acyltransferase genes in soybean (Glycine max) link: https://peerj.com/articles/14941 last-modified: 2023-03-20 description: BackgroundSoybean (Glycine max) is a major protein and vegetable oil source. In plants, diacylglycerol acyltransferase (DGAT) can exert strong flux control, which is rate-limiting for triacylglycerol biosynthesis in seed oil formation.MethodsHere, we identified soybean DGAT genes via a bioinformatics method, thereby laying a solid foundation for further research on their function. Based on our bioinformatics analyses, including gene structure, protein domain characteristics, and phylogenetic analysis, 26 DGAT putative gene family members unevenly distributed on 12 of the 20 soybean chromosomes were identified and divided into the following four groups: DGAT1, DGAT2, WS/DGAT, and cytoplasmic DGAT.ResultsThe Ka/Ks ratio of most of these genes indicated a significant positive selection pressure. DGAT genes exhibited characteristic expression patterns in soybean tissues. The differences in the structure and expression of soybean DGAT genes revealed the diversity of their functions and the complexity of soybean fatty acid metabolism. Our findings provide important information for research on the fatty acid metabolism pathway in soybean. Furthermore, our results will help identify candidate genes for potential fatty acid-profile modifications to improve soybean seed oil content.ConclusionsThis is the first time that in silico studies have been used to report the genomic and proteomic characteristics of DGAT in soybean and the effect of its specific expression on organs, age, and stages. creator: Shihui Zhao creator: Fan Yan creator: Yajing Liu creator: Monan Sun creator: Ying Wang creator: Jingwen Li creator: Xinsheng Zhang creator: Xuguang Yang creator: Qingyu Wang uri: https://doi.org/10.7717/peerj.14941 license: https://creativecommons.org/licenses/by-nc/4.0 rights: ©2023 Zhao et al. title: Development of a new generation of miniemulsion based on cottonseed oil with α-tocopherol and ZnO and evaluation of its adjuvant activity link: https://peerj.com/articles/14981 last-modified: 2023-03-20 description: BackgroundEmulsions have been widely used as immunological adjuvants. But the use of materials derived from plants such as cottonseed oil, alpha-tocopherol, or minerals such as zinc, as well as their use at the nanometric scale has been little explored. In this study, we develop a new miniemulsion and evaluated its antioxidant and phagocytic capacity, as well as parameters related to immune response stimulation by cytokine expression and antibodies production in a mice model.MethodsFormulated CN (cottonseed oil miniemulsion) and CNZ (cottonseed oil miniemulsion whit zinc oxide nanoparticles) miniemulsions were characterized by scanning electronic microscopy SEM, DLS and FT-IR. In murine macrophages, splenocytes and thymocytes primary cultures safety and cytotoxicity were determined by MTT. In macrophages the antioxidant and phagocytic capacity was evaluated. In BALB/c mice, the stimulation of the immune system was determined by the expression of cytokines and the production of antibodies.ResultsThe CN and CNZ presented stability for 90 days. Immediately after preparation, the CN presented a higher particle size (543.1 nm) than CNZ (320 nm). FT-IR demonstrated the correct nanoparticle synthesis by the absence of sulfate groups. CN and CNZ (1.25 to 10 µL/mL) had no toxic effect on macrophages (p = 0.108), splenocytes (p = 0.413), and thymocytes (p = 0.923). All CN and CNZ doses tested induced nitric oxide and antioxidants production in dose dependent manner when compared with control. CN-ovalbumin and CNZ-ovalbumin treatments in femoral subcutaneous tissue area showed inflammation with higher leukocyte infiltration compared with FCA. The intraperitoneal administration with CN, CNZ, and FCA showed a higher total intraperitoneal cells recruitment (CD14+) after 24 h of inoculation than control (p = 0.0001). CN and CNZ increased the phagocyte capacity with respect to untreated macrophages in the Candida albicans-phagocytosis assay. The evaluation of residual CFU indicated that only CN significantly decreased (p = 0.004) this value at 3 h. By other side, only CN increased (p = 0.002) the nitric oxide production. CNZ stimulated a major INFγ secretion compared with FCA at day 7. A major IL-2 secretion was observed at days 7 and 14, stimulated with CN and CNZ. Both miniemulsions did not affect the antibody isotypes production (IgG1, IgG2a, IgG3, IgA and IgM) at days 7, 14, 28, and 42. CN induced a significant IgG production against OVA, but lesser than FCA.ConclusionsThe two new miniemulsions with adjuvant and antioxidant capacity, were capable of generating leukocyte infiltration and increased cytokines and antibodies production. creator: Gustavo Sobrevilla-Hernández creator: Moisés Armides Franco-Molina creator: Diana G. Zárate-Triviño creator: Jorge R. Kawas creator: Sara Paola Hernández-Martínez creator: Paola Leonor García-Coronado creator: Silvia Elena Santana-Krímskaya creator: Cynthia Aracely Alvizo-Báez creator: Cristina Rodríguez-Padilla uri: https://doi.org/10.7717/peerj.14981 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Sobrevilla-Hernández et al. title: Responses of maize hybrids to water stress conditions at different developmental stages: accumulation of reactive oxygen species, activity of enzymatic antioxidants and degradation in kernel quality traits link: https://peerj.com/articles/14983 last-modified: 2023-03-20 description: Sustainable maize production under changing climatic conditions, especially heat and water stress conditions is one of the key challenges that need to be addressed immediately. The current field study was designed to evaluate the impact of water stress on morpho-physiological, biochemical, reactive oxygen species, antioxidant activity and kernel quality traits at different plant growth stages in maize hybrids. Four indigenous i.e., YH-5427, YH-5482, YH-5395, JPL-1908, and one multinational maize hybrid i.e., NK-8441 (Syngenta Seeds) were used for the study. Four stress treatments (i) Control (ii) 3-week water stress at pre-flowering stage (iii) 3-week water stress at anthesis stage (iv) 3-week water stress at grain filling/post-anthesis stage. The presence of significant oxidative stress was revealed by the overproduction of reactive oxygen species (ROXs) i.e., H2O2 (1.9 to 5.8 µmole g−1 FW) and malondialdehyde (120.5 to 169.0 nmole g−1 FW) leading to severe negative impacts on kernel yield. Moreover, a severe reduction in photosynthetic ability (50.6%, from 34.0 to 16.8 µmole m−2 s−1), lower transpirational rate (31.3%, from 3.2 to 2.2 mmol m−2 s−1), alterations in plant anatomy, reduced pigments stability, and deterioration of kernel quality was attributed to water stress. Water stress affected all the three studied growth stages, the pre-flowering stage being the most vulnerable while the post-anthesis stage was the least affected stage to drought stress. Antioxidant activity was observed to increase under all stress conditions in all maize hybrids, however, the highest antioxidant activity was recorded at the anthesis stage and in maize hybrids YH-5427 i.e., T-SOD activity was increased by 61.3% from 37.5 U mg−1 pro to 60.5 U mg−1 pro while CAT activity was maximum under water stress conditions 8.3 U mg−1 pro as compared to 10.3 U mg−1 pro under control (19.3%). The overall performance of maize hybrid YH-5427 was much more promising than other hybrids, attributed to its higher photosynthetic activity, and better antioxidant defense mechanism. Therefore, this hybrid could be recommended for cultivation in drought-prone areas. creator: Muhammad Irfan Yousaf creator: Muhammad Waheed Riaz creator: Aamar Shehzad creator: Shakra Jamil creator: Rahil Shahzad creator: Shamsa Kanwal creator: Aamir Ghani creator: Farman Ali creator: Muhammad Abdullah creator: Muhammad Ashfaq creator: Quaid Hussain uri: https://doi.org/10.7717/peerj.14983 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Yousaf et al. title: RNA sequencing-based exploration of the effects of far-red light on microRNAs involved in the shade-avoidance response of D. officinale link: https://peerj.com/articles/15001 last-modified: 2023-03-20 description: Dendrobium officinale (D. officinale) has remarkable medicinal functions and high economic value. The shade-avoidance response to far-red light importantly affects the D. officinale productivity. However, the regulatory mechanism of miRNAs involved in the far-red light-avoidance response is unknown. Previous studies have found that, in D. officinale, 730 nm (far-red) light can promote the accumulation of plant metabolites, increase leaf area, and accelerate stem elongation. Here, the effects of far-red light on D. officinale were analysed via RNA-seq. KEGG analysis of miRNA target genes revealed various far-red light response pathways, among which the following played central roles: the one-carbon pool by folate; ascorbate and aldarate; cutin, suberine and wax biosynthesis; and sulfur metabolism. Cytoscape analysis of DE miRNA targets showed that novel_miR_484 and novel_miR_36 were most likely involved in the effects of far-red light on the D. officinale shade avoidance. Content verification revealed that far-red light promotes the accumulation of one-carbon compounds and ascorbic acid. Combined with qPCR validation results, the results showed that miR395b, novel_miR_36, novel_miR_159, novel_miR_178, novel_miR_405, and novel_miR_435 may participate in the far-red light signalling network through target genes, regulating the D. officinale shade avoidance. These findings provide new ideas for the efficient production of D. officinale. creator: Yifan Yang creator: Yuqiang Qiu creator: Wei Ye creator: Gang Sun creator: Hansheng Li uri: https://doi.org/10.7717/peerj.15001 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Yang et al. title: Advanced polymeric metal/metal oxide bionanocomposite using seaweed Laurencia dendroidea extract for antiprotozoal, anticancer, and photocatalytic applications link: https://peerj.com/articles/15004 last-modified: 2023-03-20 description: BackgroundBiosynthesized nanoparticles are gaining popularity due to their distinctive biological applications as well as bioactive secondary metabolites from natural products that contribute in green synthesis.MethodologyThis study reports a facile, ecofriendly, reliable, and cost-effective synthesis of silver nanoparticles (AgNPs), copper oxide nanoparticles (CuONPs), and polymeric PVP-silver-copper oxide nanocomposite using ethanol extract of seaweed Laurencia dendroidea and were evaluated for antiprotozoal, anticancer and photocatalytic potential. The nanostructures of the AgNPs, CuONPs, and polymeric PVP-Ag-CuO nanocomposite were confirmed by different spectroscopic and microscopic procedures.ResultsThe UV-vis spectrum displayed distinct absorption peaks at 440, 350, and 470 nm for AgNPs, CuONPs, and polymeric Ag-CuO nanocomposite, respectively. The average particles size of the formed AgNPs, CuONPs, and Ag-CuO nanocomposite was 25, 28, and 30 nm, respectively with zeta potential values −31.7 ± 0.6 mV, −17.6 ± 4.2 mV, and −22.9 ± 4.45 mV. The microscopic investigation of biosynthesized nanomaterials revealed a spherical morphological shape with average crystallite sizes of 17.56 nm (AgNPs), 18.21 nm (CuONPs), and 25.46 nm (PVP-Ag-CuO nanocomposite). The antiprotozoal potential of green synthesized nanomaterials was examined against Leishmania amazonensis and Trypanosoma cruzi parasites. The polymeric PVP-Ag-CuO nanocomposite exerted the highest antiprotozoal effect with IC50 values of 17.32 ± 1.5 and 17.48 ± 4.2 µM, in contrast to AgNPs and CuONPs. The anticancer potential of AgNPs, CuONPs, and polymeric PVP-Ag-CuO nanocomposite against HepG2 cancer cell lines revealed that all the nanomaterials were effective and the highest anticancer potential was displayed by PVP-Ag-CuO nanocomposite with IC50 values 91.34 µg mL−1 at 200 µg mL−1 concentration. Additionally, PVP-Ag-CuO nanocomposite showed strong photocatalytic effect.ConclusionOverall, this study suggested that the biogenic synthesized nanomaterials AgNPs, CuONPs, and polymeric PVP-Ag-CuO nanocomposite using ethanol extract of seaweed L. dendroidea possesses promising antiprotozoal anticancer and photocatalytic effect and could be further exploited for the development of antiprotozoal and anticancer therapeutics agents. creator: Musarat Amina creator: Nawal M. Al Musayeib creator: Seham Alterary creator: Maha F. El-Tohamy creator: Samira A. Alhwaiti uri: https://doi.org/10.7717/peerj.15004 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Amina et al. title: Spatial heterogeneity in the exclusive use of hygienic materials during menstruation among women in urban India link: https://peerj.com/articles/15026 last-modified: 2023-03-20 description: BackgroundMenstrual hygiene is essential for women to live with dignity. However, a large proportion of Indian women still suffer from unhygienic menstrual practices leading to reproductive tract infections. To understand the socioeconomic and bio-demographic determinants of menstrual hygiene practices, various national or local level studies have been conducted in India and around the world, however, no previous study has tried to understand the spatial heterogeneity across Indian districts in the use of hygienic materials among young urban women.MethodsThis study used data from 54,561 urban women aged 15–24 from the National Family Health Survey-5. Global Moran’s I was applied to assess the degree of spatial autocorrelation and cluster and outlier analyses to locate hot-spots and clod-spots in the exclusive use of hygienic materials across the districts. Ordinary least square, spatial lag, and error models were used to identify determinants of exclusive use of hygienic materials.ResultsApproximately 66.8% of urban women exclusively use of hygienic materials which varied across districts. Global Moran’s I of 0.46 indicated positive spatial autocorrelation in the outcome. Cluster and outlier analysis revealed cold-spots in central Indian districts and hotspots in south Indian districts. Results of spatial error model identified women’s years of schooling, marital status, social group, and household wealth were major determinants of the exclusive use of hygienic materials among urban women across Indian districts.ConclusionSubstantial spatial heterogeneity in the outcome among urban women in India suggests the need to design targeted and context-specific behavioural interventions and programs for women in urban India. creator: Aditya Singh creator: Mahashweta Chakrabarty uri: https://doi.org/10.7717/peerj.15026 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Singh et al. title: Pesticides reduce tropical amphibian and reptile diversity in agricultural landscapes in Indonesia link: https://peerj.com/articles/15046 last-modified: 2023-03-20 description: Pesticide use on tropical crops has increased substantially in recent decades, posing a threat to biodiversity and ecosystem services. Amphibians and reptiles are common in tropical agricultural landscapes, but few field studies measure pesticide impacts on these taxa. Here we combine 1-year of correlative data with an experimental field approach from Indonesia. We show that while pesticide application cannot predict amphibian or reptile diversity patterns in cocoa plantations, our experimental exposure to herbicides and insecticides in vegetable gardens eliminated amphibians, whereas reptiles were less impacted by insecticide and not affected by herbicide exposure. The pesticide-driven loss of a common amphibian species known to be a pest-control agent (mainly invertebrate predation) suggests a strong indirect negative effect of pesticides on this service. We recommend landscape-based Integrated Pest Management and additional ecotoxicological studies on amphibians and reptiles to underpin a regulatory framework and to assure recognition and protection of their ecosystem services. creator: Thomas Cherico Wanger creator: Barry W. Brook creator: Theodore Evans creator: Teja Tscharntke uri: https://doi.org/10.7717/peerj.15046 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Wanger et al. title: GOLPH3 modulates expression and alternative splicing of transcription factors associated with endometrial decidualization in human endometrial stromal cells link: https://peerj.com/articles/15048 last-modified: 2023-03-20 description: Endometrial decidualization is a decidual tissue formed by the proliferation and re-differentiation of endometrial stroma stimulated by decidualization inducing factors. It is very important for the proper maintenance of pregnancy. Previous studies speculated that Golgi phosphoprotein 3 (GOLPH3) may have a regulatory role in the process of endometrial decidualization, while the specific molecular mechanisms of GOLPH3 is unclear. In this part, GOLPH3 was silenced in human endometrial stromal cells (hESCs), and the transcriptome data (RNA-seq) by GOLPH3 knockdown (siGOLPH3) was obtained by high-throughput sequencing technology so as to analyze the potential targets of GOLPH3 at expression and alternative splicing levels in hESCs. Through bioinformatics analysis, we found that siGOLPH3 can significantly affect the overall transcriptional level of hESCs. A total of 6,025 differentially expressed genes (DEGs) and 4,131 differentially alternative splicing events (DASEs) were identified. Through functional cluster analysis of these DEGs and genes where differential alternative splicing events are located, it is found that they are enriched in the PI3K/Akt signaling pathway, RNA splicing and processing, transcription factors and other pathways related to endometrial decidualization and important biological processes, indicating the important biological function of GOLPH3. At the same time, we focused on the analysis of the transcription factors regulated by GOLPH3, including gene expression regulation and the regulation of variable splicing. We found that GOLPH3can regulate the expression of transcription factors such as LD1, FOSL2, GATA2, CSDC2 and CREB3L1. At the same time, it affects the variable splicing mode of FOXM1 and TCF3. The function of these transcription factors is directly related to decidualization of endometrium. Therefore, we infer that GOLPH3 may participate in endometrial de membrane by regulating expression and alternative splicing levels of transcription factors. We further identified the role of GOLPH3 in the transcriptional mechanism. At the same time, it also expands the function mode of GOLPH3 protein molecule, and provides a theoretical basis for downstream targeted drug research and development and clinical application. creator: Suqin Zhu creator: Dianliang Lin creator: Zhoujie Ye creator: Xiaojing Chen creator: Wenwen Jiang creator: Huiling Xu creator: Song Quan creator: Beihong Zheng uri: https://doi.org/10.7717/peerj.15048 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Zhu et al. title: Artificial shelters and marine infectious disease: no detectable effect of the use of casitas to enhance juvenile Panulirus argus in shelter-poor habitats on a viral disease dynamics link: https://peerj.com/articles/15073 last-modified: 2023-03-20 description: Casitas, low-lying artificial shelters that mimic large crevices, are used in some fisheries for Caribbean spiny lobsters (Panulirus argus). These lobsters are highly gregarious and express communal defense of the shelter. Scaled-down casitas have been shown to increase survival, persistence, and foraging ranges of juveniles. Therefore, the use of casitas has been suggested to help enhance local populations of juvenile P. argus in Caribbean seagrass habitats, poor in natural crevice shelters, in marine protected areas. Following the emergence of Panulirus argus virus 1 (PaV1), which is lethal to juveniles of P. argus, concern was raised about the potential increase in PaV1 transmission with the use of casitas. It was then discovered that lobsters tend to avoid shelters harboring diseased conspecifics, a behavior which, alone or in conjunction with predatory culling of diseased lobsters, has been proposed as a mechanism reducing the spread of PaV1. However, this behavior may depend on the ecological context (i.e., availability of alternative shelter and immediacy of predation risk). We conducted an experiment in a lobster nursery area to examine the effect of the use of casitas on the dynamics of the PaV1 disease. We deployed 10 scaled-down casitas per site on five 1-ha sites over a reef lagoon (casita sites) and left five additional sites with no casitas (control sites). All sites were sampled 10 times every 3–4 months. Within each site, all lobsters found were counted, measured, and examined for clinical signs of the PaV1 disease. Mean density and size of lobsters significantly increased on casita sites relative to control sites, but overall prevalence levels remained similar. There was no relationship between lobster density and disease prevalence. Dispersion parameters (m and k of the negative binomial distribution) revealed that lobsters tended to avoid sharing natural crevices, but not casitas, with diseased conspecifics. These results confirm that casitas provide much needed shelter in seagrass habitats and that their large refuge area may allow distancing between healthy and diseased lobsters. On eight additional sampling times over two years, we culled all diseased lobsters observed on casita sites. During this period, disease prevalence did not decrease but rather increased and varied with site, suggesting that other factors (e.g., environmental) may be influencing the disease dynamics. Using scaled-down casitas in shelter-poor habitats may help efforts to enhance juvenile lobsters for conservation purposes, but monitoring PaV1 prevalence at least once a year during the first few years would be advisable. creator: Rebeca Candia-Zulbarán creator: Patricia Briones-Fourzán creator: Fernando Negrete-Soto creator: Cecilia Barradas-Ortiz creator: Enrique Lozano-Álvarez uri: https://doi.org/10.7717/peerj.15073 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Candia-Zulbarán et al. title: Wastewater monitoring of SARS-CoV-2 RNA at K-12 schools: comparison to pooled clinical testing data link: https://peerj.com/articles/15079 last-modified: 2023-03-20 description: BackgroundWastewater measurements of SARS-CoV-2 RNA have been extensively used to supplement clinical data on COVID-19. Most examples in the literature that describe wastewater monitoring for SARS-CoV-2 RNA use samples from wastewater treatment plants and individual buildings that serve as the primary residence of community members. However, wastewater surveillance can be an attractive supplement to clinical testing in K-12 schools where individuals only spend a portion of their time but interact with others in close proximity, increasing risk of potential transmission of disease.MethodsWastewater samples were collected from two K-12 schools in California and divided into solid and liquid fractions to be processed for detection of SARS-CoV-2. The resulting detection rate in each wastewater fraction was compared to each other and the detection rate in pooled clinical specimens.ResultsMost wastewater samples were positive for SARS-CoV-2 RNA when clinical testing was positive (75% for solid samples and 100% for liquid samples). Wastewater samples continued to test positive for SARS-CoV-2 RNA when clinical testing was negative or in absence of clinical testing (83% for both solid and liquid samples), indicating presence of infected individuals in the schools. Wastewater solids had a higher concentration of SARS-CoV-2 than wastewater liquids on an equivalent mass basis by three orders of magnitude. creator: Sooyeol Kim creator: Alexandria B. Boehm uri: https://doi.org/10.7717/peerj.15079 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Kim and Boehm title: Measurement error associated with gait cycle selection in treadmill running at various speeds link: https://peerj.com/articles/14921 last-modified: 2023-03-17 description: A common approach in the biomechanical analysis of running technique is to average data from several gait cycles to compute a ‘representative mean.’ However, the impact of the quantity and selection of gait cycles on biomechanical measures is not well understood. We examined the effects of gait cycle selection on kinematic data by: (i) comparing representative means calculated from varying numbers of gait cycles to ‘global’ means from the entire capture period; and (ii) comparing representative means from varying numbers of gait cycles sampled from different parts of the capture period. We used a public dataset (n = 28) of lower limb kinematics captured during a 30-second period of treadmill running at three speeds (2.5 m s−1, 3.5 m s−1 and 4.5 m s−1). ‘Ground truth’ values were determined by averaging data across all collected strides and compared to representative means calculated from random samples (1,000 samples) of n (range = 5–30) consecutive gait cycles. We also compared representative means calculated from n (range = 5–15) consecutive gait cycles randomly sampled (1,000 samples) from within the same data capture period. The mean, variance and range of the absolute error of the representative mean compared to the ‘ground truth’ mean progressively reduced across all speeds as the number of gait cycles used increased. Similar magnitudes of ‘error’ were observed between the 2.5 m s−1 and 3.5 m s−1 speeds at comparable gait cycle numbers —where the maximum errors were < 1.5 degrees even with a small number of gait cycles (i.e., 5–10). At the 4.5 m s−1 speed, maximum errors typically exceeded 2–4 degrees when a lower number of gait cycles were used. Subsequently, a higher number of gait cycles (i.e., 25–30) was required to achieve low errors (i.e., 1–2 degrees) at the 4.5 m s−1 speed. The mean, variance and range of absolute error of representative means calculated from different parts of the capture period was consistent irrespective of the number of gait cycles used. The error between representative means was low (i.e., < 1.5 degrees) and consistent across the different number of gait cycles at the 2.5 m s−1 and 3.5 m s−1 speeds, and consistent but larger (i.e., up to 2–4 degrees) at the 4.5 m s−1 speed. Our findings suggest that selecting as many gait cycles as possible from a treadmill running bout will minimise potential ‘error.’ Analysing a small sample (i.e., 5–10 cycles) will typically result in minimal ‘error’ (i.e., < 2 degrees), particularly at lower speeds (i.e., 2.5 m s−1 and 3.5 m s−1). Researchers and clinicians should consider the balance between practicalities of collecting and analysing a smaller number of gait cycles against the potential ‘error’ when determining their methodological approach. Irrespective of the number of gait cycles used, we recommend that the potential ‘error’ introduced by the choice of gait cycle number be considered when interpreting the magnitude of effects in treadmill-based running studies. creator: Aaron S. Fox creator: Jason Bonacci creator: John Warmenhoven creator: Meghan F. Keast uri: https://doi.org/10.7717/peerj.14921 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Fox et al. title: Current trends in nano-delivery systems for functional foods: a systematic review link: https://peerj.com/articles/14980 last-modified: 2023-03-17 description: BackgroundIncreased awareness of the relationship between certain components in food beyond basic nutrition and health has generated interest in the production and consumption. Functional foods owe much of their health benefits to the presence of bioactive components. Despite their importance, their poor stability, solubility, and bioavailability may require the use of different strategies including nano-delivery systems (NDS) to sustain delivery and protection during handling, storage, and ingestion. Moreover, increasing consumer trend for non-animal sourced ingredients and interest in sustainable production invigorate the need to evaluate the utility of plant-based NDS.MethodIn the present study, 129 articles were selected after screening from Google Scholar searches using key terms from current literature.ScopeThis review provides an overview of current trends in the use of bioactive compounds as health-promoting ingredients in functional foods and the main methods used to stabilize these components. The use of plant proteins as carriers in NDS for bioactive compounds and the merits and challenges of this approach are also explored. Finally, the review discusses the application of protein-based NDS in food product development and highlights challenges and opportunities for future research.Key FindingsPlant-based NDS is gaining recognition in food research and industry for their role in improving the shelf life and bioavailability of bioactives. However, concerns about safety and possible toxicity limit their widespread application. Future research efforts that focus on mitigating or enhancing their safety for food applications is warranted. creator: Emmanuel Otchere creator: Brighid M. McKay creator: Marcia M. English creator: Alberta N. A. Aryee uri: https://doi.org/10.7717/peerj.14980 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Otchere et al. title: Size-age population structure of an endangered and anthropogenically introgressed northern Adriatic population of marble trout (Salmo marmoratus Cuv.): insights for its conservation and sustainable exploitation link: https://peerj.com/articles/14991 last-modified: 2023-03-17 description: Salmonid species are main actors in the Italian socio-ecological landscape of inland fisheries. We present novel data on the size-age structure of one of the remnant Italian populations of the critically endangered marble trout Salmo marmoratus, which co-occurs with other stocked non-native salmonids in a large glacial river of the Lake Maggiore basin (Northern Italy-Southern Switzerland). Like other Italian native trout populations, the Toce River marble trout population is affected by anthropogenic introgression with the non-native brown trout S. trutta. Our sample includes 579 individuals, mainly collected in the Toce River main channel. We estimated the length-weight relationship, described the population size-age structure, estimated the age-specific growth trajectories, and fit an exponential mortality model. A subset of the sample was also used to measure numerical and biomass density. The estimated asymptotic maximum length is ~105 cm total length (TL). Mean length at first maturity is ~55 cm TL, and mean length at maximum yield per recruit is ~68 cm TL. Approximately 45–70% of the population are estimated to die annually, along with a fishing annual mortality of ~37%, with an exploitation ratio of ~0.5. The frequency distribution of length classes in a sample collected by angling shows that ~80% of the individuals that could be retained according to the current recreational fishing regulations likely never reproduced, and large fish disproportionally contributing to recruitment are fished and retained. We identify possible overfishing risks posed by present regulations, and propose updated harvest-slot length limits to mitigate such risks. More detailed and long-term datasets on this system are needed to more specifically inform the fishery management and monitor the effects of any change in the management strategy on the size-age structure of the marble trout population of the Toce River. creator: Gianluca Polgar creator: Mattia Iaia creator: Paolo Sala creator: Tsung Fei Khang creator: Silvia Galafassi creator: Silvia Zaupa creator: Pietro Volta uri: https://doi.org/10.7717/peerj.14991 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Polgar et al. title: Clinical performance of zirconium implants compared to titanium implants: a systematic review and meta-analysis of randomized controlled trials link: https://peerj.com/articles/15010 last-modified: 2023-03-17 description: PurposeTo quantitatively assess and compare the clinical outcomes, including survival rate, success rate, and peri-implant indices of titanium and zirconium implants in randomized controlled trials.MethodsThe electronic databases searched included the Cochrane Central Register of Controlled Trials (CENTRAL), Medline via Ovid, EMBASE, and Web of Science. Randomized controlled trials (RCTs) that reported the effects of zirconium implants on primary outcomes, such as survival rate, success rate, marginal bone loss (MBL), and probing pocket depth (PPD), compared to titanium implants were included in this review. Two reviewers independently screened and selected the records, assessed their quality, and extracted the data from the included studies.ResultsA total of four studies from six publications reviewed were included. Two of the comparative studies were assessed at minimal risk of bias. Zirconium implants may have a lower survival rate (risk ratio (RR) = 0.91, CI [0.82–1.02], P = 0.100, I2 = 0%) and a significantly lower success rate than titanium implants (RR = 0.87, CI [0.78–0.98], P = 0.030, I2 = 0%). In addition, there was no difference between the titanium and zirconium implants in terms of MBL, PPD, bleeding on probing (BOP), plaque index (PI), and pink esthetic score (PES) (for MBL, MD = 0.25, CI [0.02–0.49], P = 0.033, I2 = 0%; for PPD, MD = −0.07, CI [−0.19–0.05], P = 0.250, I2 = 31%).ConclusionZirconium implants may have higher failure rates due to their mechanical weakness. Zirconium implants should be strictly assessed before they enter the market. Further studies are required to confirm these findings. creator: Chengchen Duan creator: Li Ye creator: Mengyun Zhang creator: Lei Yang creator: Chunjie Li creator: Jian Pan creator: Yingying Wu creator: Yubin Cao uri: https://doi.org/10.7717/peerj.15010 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Duan et al. title: Roles of cuproptosis-related gene DLAT in various cancers: a bioinformatic analysis and preliminary verification on pro-survival autophagy link: https://peerj.com/articles/15019 last-modified: 2023-03-17 description: BackgroundStudies have shown that the expressions and working mechanisms of Dihydrolipoamide S-acetyltransferase (DLAT) in different cancers vary. It is necessary to analyze the expressions and regulatory roles of DLAT in tumors systematically.MethodsOnline public-platform literature on the relationships between DLAT expression levels and tumor prognosis, methylation status, genetic alteration, drug sensitivity, and immune infiltration has been reviewed. The literature includes such documents as The Cancer Genome Atlas (TCGA), Human Protein Atlas (HPA), Tumor Immune Estimation Resource 2.0 (TIMER2.0), Gene Expression Profiling Interactive Analysis 2 (GEPIA2) and Receiver Operating Characteristic plotter (ROC plotter). The molecular mechanisms of DLAT were explored with the Gene Set Enrichment Analysis (GSEA). The relationship between down-regulated DLAT and autophagy in two liver hepatocellular carcinoma (LIHC) cell lines was confirmed with the western blot method, colony formation assay, and transmission electron microscopy. Tissue microarrays were validated through the immunohistochemical staining of DLAT.ResultsDLAT is upregulated in the LIHC, lung adenocarcinoma (LUAD), lung squamous cell carcinoma (LUSC), and stomach adenocarcinoma (STAD) tumors but is down-regulated in the head and neck squamous cell carcinoma (HNSC) and kidney renal clear cell carcinoma (KIRC) tumors in comparison with normal tissues. For LIHC patients treated with 5-Fluorouracil and Lenvatinib, the DLAT levels of those in the drug-resistant group are significantly high. In LIHC cells, autophagy will be inhibited, and cell death will be induced when DLAT breaks down. Moreover, there exist positive correlations between DLAT expression levels and infiltration of B cells, DC cells, Tregs, and CD8+ T cells in kidney chromophobe (KICH), breast invasive carcinoma (BRCA), prostate adenocarcinoma (PRAD), LIHC and HPV+ HNSC. In LIHC, markers of Tregs are positively correlated with DLAT. Compared with those of normal tissues, the staining intensity of DLAT and the amount of Tregs marker CD49d in LIHC increase.ConclusionsThrough this study, the expressions of DLAT in various cancer types can be understood comprehensively. It suggests that DLAT may be a prognostic marker for LIHC, LUAD, LUSC, STAD and KIRC. A high DLAT expression in LIHC may promote tumorigenesis by stimulating autophagy and inhibiting anti-tumor immunity. creator: Qinjing Yang creator: Shuangshuang Zeng creator: Wei Liu uri: https://doi.org/10.7717/peerj.15019 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Yang et al. title: A prospective follow-up of thyroid volume and thyroiditis features on ultrasonography among survivors of predominantly mild to moderate COVID-19 link: https://peerj.com/articles/15034 last-modified: 2023-03-17 description: BackgroundWe previously showed that higher SARS-CoV-2 viral load correlated with smaller thyroid volumes among COVID-19 survivors at 2 months after acute COVID-19. Our current follow-up study evaluated the evolution of thyroid volumes and thyroiditis features within the same group of patients 6 months later.MethodsAdult COVID-19 survivors who underwent thyroid ultrasonography 2 months after infection (USG1) were recruited for follow-up USG 6 months later (USG2). The primary outcome was the change in thyroid volume. We also reassessed thyroiditis features on USG, thyroid function and anti-thyroid antibodies.ResultsFifty-four patients were recruited (mean age 48.1 years; 63% men). The mean thyroid volume increased from USG1 to USG2 (11.9 ± 4.8 to 14.5 ± 6.2 mL, p < 0.001). Thirty-two patients (59.3%) had significant increase in thyroid volume by ≥15%, and they had a median increase of +33.3% (IQR: +20.0% to +45.0%). Multivariable logistic regression analysis showed that only higher baseline SARS-CoV-2 viral load independently correlated with significant thyroid volume increase on USG2 (p = 0.022). Among the seven patients with thyroiditis features on USG1, six (85.7%) had the features resolved on USG2. None had new thyroiditis features on USG2. All abnormal thyroid function during acute COVID-19 resolved upon USG1 and USG2.ConclusionMost COVID-19 survivors had an increase in thyroid volume from early convalescent phase to later convalescent phase. This increase correlated with high initial SARS-CoV-2 viral load. Together with the resolution of thyroiditis features, these may suggest a transient direct atrophic effect of SARS-CoV-2 on the thyroid gland with subsequent recovery of thyroid volume and thyroiditis features. creator: Man Him Matrix Fung creator: David Tak Wai Lui creator: Keith Wan Hang Chiu creator: Sherman Haynam Lee creator: Chi Ho Lee creator: Wing Sun Chow creator: Alan Chun Hong Lee creator: Anthony Raymond Tam creator: Polly Pang creator: Tip Yin Ho creator: Carol Ho Yi Fong creator: Connie Hong Nin Loong creator: Chun Yiu Law creator: Kelvin Kai Wang To creator: Ching Wan Lam creator: Kathryn Choon Beng Tan creator: Yu Cho Woo creator: Ivan Fan Ngai Hung creator: Karen Siu Ling Lam creator: Brian Lang uri: https://doi.org/10.7717/peerj.15034 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Fung et al. title: Effects of vegetation restoration on distribution characteristics of heavy metals in soil in Karst plateau area of Guizhou link: https://peerj.com/articles/15044 last-modified: 2023-03-17 description: In southwest China, vegetation restoration is widely used in karst rocky desertification control projects. This technology can effectively fix the easily lost soil, gradually restore the plant community and improve soil fertility. However, the change law of soil heavy metals in the restoration process remains to be further studied. Therefore, in this work, Guizhou Caohai Nature Reserve as a typical karst area was taken as the research object to investigate the influence of vegetation restoration technology on repairing soil heavy metal pollution. The spatial distribution characteristics of soil heavy metals (chromium, nickel, arsenic, zinc, lead) before and after vegetation restoration in karst area were studied by comparative analysis and linear stepwise regression analysis. The main influencing factors and spatial distribution characteristics of heavy metals in karst area were further discussed. The results showed that: (1) heavy metals in karst soils are affected by surface vegetation, root exudates, microorganisms and leaching. Only heavy metals nickel (Ni) and lead (Pb) showed the tendency of surface enrichment and bottom precipitation enrichment in non-karst soils. Path analysis suggested that non-metallic soil factors such as soil bulk density (BD), total nitrogen (TN) and ammonium nitrogen (NH4+-N) had direct effect on the content of heavy metals in soil. (2) The proportion of 0.25–2 mm aggregates in the surface soil of vegetation restoration belt was more than 40%, and the proportion of surface soil ≤2 mm aggregates in this increased to 83% and 88%, respectively, which could improve the soil structure and properties effectively. (3) Vegetation restoration effectively restored the nutrient elements such as carbon and nitrogen in the soil, and enhanced the soil material circulation. Furthermore the content of heavy metals in the surface soil higher than that in the 10–20 cm soil layer. Plant absorption, biosorption mechanism of microorganisms, coupling of root exudates, dissolution of soil soluble organic carbon and pH make the contents of heavy metals Cr, Ni and Pb in vegetation restoration belt slightly lower than those in karst soil. At the same time, affected by vegetation coverage, residual heavy metals in soil are further leached by surface runoff. Therefore, the content of heavy metals in soil could reduce combined heavy metal enrichment plants for extraction with remediation. This study elucidates the advantages and remedy mechanism of vegetation restoration in the remediation of heavy metal contaminated soils in Caohai area of Guizhou, and this plant activation and enrichment extraction remediation technology would be popularized and applied in the remediation of heavy metal contaminated soils in other karst areas. creator: Yunjie Wu creator: Xin Tian creator: Runze Wang creator: Mingyi Zhang creator: Shuo Wang uri: https://doi.org/10.7717/peerj.15044 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Wu et al. title: Ferroptosis-dependent breast cancer cell-derived exosomes inhibit migration and invasion of breast cancer cells by suppressing M2 macrophage polarization link: https://peerj.com/articles/15060 last-modified: 2023-03-17 description: AimFerroptosis, a novel type of iron-dependent cell death, plays a vital role in breast cancer progression. However, the function of ferroptosis-induced cancer cell-derived exosomes in breast cancer remains unclear. In this study, we attempted to investigate the impact of breast cancer cells-derived exosomes induced by ferroptosis on the polarization of macrophages and the progression of breast cancer.MethodsErastin was used to induce ferroptosis and breast cancer cell-derived exosomes were identified by transmission electron microscopy. Western blot, quantitative reverse transcription PCR, immunofluorescence, flow cytometry, and ELISA were used to determine the role of exosomes in macrophage polarization. Transwell assays were used to detect breast cancer cell migration, and invasion.ResultsOur results showed that erastin promoted ferroptosis in breast cancer cells with increased Fe2+ level and ROS production. Breast cancer cell-derived exosomes induced by ferroptosis were successfully isolated and verified to be internalized by macrophages. In addition, ferroptosis-induced breast cancer cell-derived exosomes (Fe-exo) remarkably diminished M2 marker, Arg-1 expression. The ratio of CD206+ macrophages was significantly decreased after Fe-exo treatment. CD206 protein expression and Arg-1 level were dramatically reduced in M2 macrophages incubated by Fe-exo. Moreover, autophagy PCR array showed that the expression of 84 autophagy-related genes were altered after macrophages were incubated by Fe-exo. Furthermore, macrophages incubated by Fe-exo repressed the migration and invasion of breast cancer cells.ConclusionFerroptosis-dependent cancer cell-derived exosomes inhibited M2 polarization of macrophages, which in turn inhibited migration and invasion of breast cancer cells. This study provides novel therapeutic strategies for patients with breast cancer. creator: Chenghao Yi creator: Shilong Wu creator: Qianyu Duan creator: Lei Liu creator: Li Li creator: Yonghui Luo creator: Anan Wang uri: https://doi.org/10.7717/peerj.15060 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Yi et al. title: “In the light of evolution:” keratins as exceptional tumor biomarkers link: https://peerj.com/articles/15099 last-modified: 2023-03-17 description: Keratins (KRTs) are the intermediate filament-forming proteins of epithelial cells, classified, according to their physicochemical properties, into “soft” and “hard” keratins. They have a key role in several aspects of cancer pathophysiology, including cancer cell invasion and metastasis, and several members of the KRT family serve as diagnostic or prognostic markers. The human genome contains both, functional KRT genes and non-functional KRT pseudogenes, arranged in two uninterrupted clusters on chromosomes 12 and 17. This characteristic renders KRTs ideal for evolutionary studies. Herein, comprehensive phylogenetic analyses of KRT homologous proteins in the genomes of major taxonomic divisions were performed, so as to fill a gap in knowledge regarding the functional implications of keratins in cancer biology among tumor-bearing species. The differential expression profiles of KRTs in diverse types of cancers were investigated by analyzing high-throughput data, as well. Several KRT genes, including the phylogenetically conserved ones, were found to be deregulated across several cancer types and to participate in a common protein-protein interaction network. This indicates that, at least in cancer-bearing species, these genes might have been under similar evolutionary pressure, perhaps to support the same important function(s). In addition, semantic relations between KRTs and cancer were detected through extensive text mining. Therefore, by applying an integrative in silico pipeline, the evolutionary history of KRTs was reconstructed in the context of cancer, and the potential of using non-mammalian species as model organisms in functional studies on human cancer-associated KRT genes was uncovered. creator: Işıl Takan creator: Gökhan Karakülah creator: Aikaterini Louka creator: Athanasia Pavlopoulou uri: https://doi.org/10.7717/peerj.15099 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Takan et al. title: Comparative study of convolutional neural network architectures for gastrointestinal lesions classification link: https://peerj.com/articles/14806 last-modified: 2023-03-16 description: The gastrointestinal (GI) tract can be affected by different diseases or lesions such as esophagitis, ulcers, hemorrhoids, and polyps, among others. Some of them can be precursors of cancer such as polyps. Endoscopy is the standard procedure for the detection of these lesions. The main drawback of this procedure is that the diagnosis depends on the expertise of the doctor. This means that some important findings may be missed. In recent years, this problem has been addressed by deep learning (DL) techniques. Endoscopic studies use digital images. The most widely used DL technique for image processing is the convolutional neural network (CNN) due to its high accuracy for modeling complex phenomena. There are different CNNs that are characterized by their architecture. In this article, four architectures are compared: AlexNet, DenseNet-201, Inception-v3, and ResNet-101. To determine which architecture best classifies GI tract lesions, a set of metrics; accuracy, precision, sensitivity, specificity, F1-score, and area under the curve (AUC) were used. These architectures were trained and tested on the HyperKvasir dataset. From this dataset, a total of 6,792 images corresponding to 10 findings were used. A transfer learning approach and a data augmentation technique were applied. The best performing architecture was DenseNet-201, whose results were: 97.11% of accuracy, 96.3% sensitivity, 99.67% specificity, and 95% AUC. creator: Erik O. Cuevas-Rodriguez creator: Carlos E. Galvan-Tejada creator: Valeria Maeda-Gutiérrez creator: Gamaliel Moreno-Chávez creator: Jorge I. Galván-Tejada creator: Hamurabi Gamboa-Rosales creator: Huizilopoztli Luna-García creator: Arturo Moreno-Baez creator: José María Celaya-Padilla uri: https://doi.org/10.7717/peerj.14806 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Cuevas-Rodriguez et al. title: Level of attention mediates the association between connectedness to nature and aesthetic evaluations of photographs of nature link: https://peerj.com/articles/14926 last-modified: 2023-03-16 description: Aesthetic experiences of nature are associated with beneficial psychological and behavioural outcomes. We investigated in a laboratory study whether an individual’s level of connectedness to nature is associated with their aesthetic sensitivity to images of natural scenes, and whether the amount of attention allocated to the images mediated this association. Participants (N = 82) viewed 14 photographs depicting natural scenes and evaluated them on three aesthetic dimensions and completed the Connectedness to Nature (CN) and Openness to Experience (OtE) scales. CN positively predicted pleasure, beauty and aesthetic emotion, independently of OtE. The amount of attention participants paid to the images mediated the relationship between connectedness to nature and aesthetic pleasure, and connectedness to nature and beauty ratings. These findings extend our understanding by showing that attention is an important mechanism through which nature connectedness influences aesthetic responses of pleasantness and beauty in response to natural scenes. The findings have real-world implications as appreciation of the aesthetic qualities of nature is associated with a number of beneficial psychological outcomes. creator: Neil Harrison uri: https://doi.org/10.7717/peerj.14926 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Harrison title: Structure and characteristics of the plant-frugivore bird network from the Guilin Botanical Garden link: https://peerj.com/articles/15028 last-modified: 2023-03-16 description: The interaction between plants and frugivores is crucial to ecosystem function and community diversity. However, little is known about the interaction between plants and frugivorous bird species in urban green spaces. We observed interactions between plants and frugivorous birds in the Guilin Botanical Garden for one year and determined the structure and characteristics of the interaction network. We also analyzed the impact of species traits on their network roles. Interactions between 14 frugivorous birds and 13 fruit plant species were recorded in the study area. Autumn interactions comprised 38.79% of the overall network, and winter interactions comprised 33.15%. The modularity (Q, z-score) of the network was higher in autumn; the weighted nestedness (wNODF, z-score) and interaction evenness (E2, z-score) of the network were higher in winter; the connectance (C, z-score) and interaction diversity (z-score) of the network were higher in spring; and the specialization (H2′, z-score) of the network was higher in summer. The observed network showed lower C, lower interaction H2, lower E2 , lower wNODF, higher H2′ and higher Q when compared to the random networks. The bird species most important to network stability were Hemixos castanonotus, Parus venustulus, and Pycnonotus sinensis. The most important plant species were Alocasia macrorrhiza, Cinnamomum camphora, and Machilus nanmu. Of all the bird and plant traits included in this study, only plant color had a significant impact on species strength, with black fruit having a higher species strength. Our results suggest that interaction networks in urban green spaces can be temporally complex and variable and that a network approach can be an important monitoring tool for detecting the status of crucial ecosystem functions. creator: Guohai Wang creator: Yang Huang creator: Wei Yao creator: Qiuchan Huang creator: Yongping Huang creator: Lijuan Wei creator: Qihai Zhou uri: https://doi.org/10.7717/peerj.15028 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Wang et al. title: Turkish nursing students’ compliance to standard precautions during the COVID-19 pandemic link: https://peerj.com/articles/15056 last-modified: 2023-03-16 description: ObjectivesThe aim of this study was to assess nursing students’ compliance to standard precautions during the COVID-19 pandemic.MethodsA cross-sectional study was conducted from December 2021 to June 2022, 816 nursing students participated in the study. A socio-demographic questionnaire and Compliance with Standard Precautions Scale were used to collect data. Means and percentages were used to report socio-demographic characteristics multiple regression analysis used to identify the factors influencing compliance with standard precautions.ResultsThe mean age of nursing students was 21.30 ± 1.31 years. The majority of nursing students were female, with 703 (86.2%) being female and 113 (13.8%) being male. Compliance among nursing students was 76.8% overall. Nursing students reported the highest compliance (97.7%) with putting used sharp articles into sharp boxes, with 97.1% compliance for covering mouth and nose when wearing a mask. Participants reported the lowest (38.6%) when it came to not recapping used needles after giving an injection. Regression analysis revealed that gender, year of study, and having needlestick injury or contact with blood/body fluids experience all influenced nursing students’ compliance with standard precautions.ConclusionsDuring the pandemic, nursing students compliance to standard precautions was optimal, according to this study. More research should be done to assess nursing students’ compliance with standard precautions and the effect of infection control strategies used to prevent COVID-19 transmission. creator: Sevcan Topçu creator: Zuhal Emlek Sert uri: https://doi.org/10.7717/peerj.15056 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Topçu and Emlek Sert title: Halimeda tuna (Bryopsidales, Ulvophyceae) calcification on the depth transect in the northern Adriatic Sea; carbonate production on the microscale of individual segments link: https://peerj.com/articles/15061 last-modified: 2023-03-16 description: Halimeda tuna (J. Ellis & Solander) J.V. Lamouroux is the only Halimeda species found in the Mediterranean Sea, and it is an important habitat former. In the northern Adriatic, H. tuna is among the ten most abundant seaweeds in the upper-infralittoral belt in spring and autumn. The modular thalli consist of serially arranged calcified segments. Calcification is closely related to photosynthesis, which causes alkalinization of the inter-utricular space and triggers aragonite formation. Understanding of the complex patterns of segment shape plasticity in relation to CaCO3content at different depth levels is still incomplete. Geometric morphometrics was used to investigate H. tuna segment shape variation on the depth transect at Cape Madona Nature Monument in the northern Adriatic Sea. The position on the thallus and the CaCO3 content of each studied segment were recorded, allowing slight changes in mineral content to be detected at the microscale of the segments. Our results showed that shape, size, or asymmetry of H. tuna segments were not significantly affected by depth. On the other hand, plants that grew deeper were generally more calcified. The apical and subapical segments contributed to the increase in CaCO3 content at the deeper sites, whereas the basal segments did not. This indicates that reniform or oval segments positioned apically or subapically play a key role in calcification of H. tuna in Mediterranean ecosystems. creator: Yvonne Nemcova creator: Martina Orlando-Bonaca creator: Jiri Neustupa uri: https://doi.org/10.7717/peerj.15061 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Nemcova et al. title: Concordance of chest x-ray with chest CT by body mass index link: https://peerj.com/articles/15090 last-modified: 2023-03-16 description: IntroductionPatients with suspected thoracic pathology frequently get imaging with conventional radiography or chest x-rays (CXR) and computed tomography (CT). CXR include one or two planar views, compared to the three-dimensional images generated by chest CT. CXR imaging has the advantage of lower costs and lower radiation exposure at the expense of lower diagnostic accuracy, especially in patients with large body habitus.ObjectivesTo determine whether CXR imaging could achieve acceptable diagnostic accuracy in patients with a low body mass index (BMI).MethodsThis retrospective study evaluated 50 patients with age of 63 ± 12 years old, 92% male, BMI 31.7 ± 7.9, presenting with acute, nontraumatic cardiopulmonary complaints who underwent CXR followed by CT within 1 day. Diagnostic accuracy was determined by comparing scan interpretation with the final clinical diagnosis of the referring clinician.ResultsCT results were significantly correlated with CXR results (r = 0.284, p = 0.046). Correcting for BMI did not improve this correlation (r = 0.285, p = 0.047). Correcting for BMI and age also did not improve the correlation (r = 0.283, p = 0.052), nor did correcting for BMI, age, and sex (r = 0.270, p = 0.067). Correcting for height alone slightly improved the correlation (r = 0.290, p = 0.043), as did correcting for weight alone (r = 0.288, p = 0.045). CT accuracy was 92% (SE = 0.039) vs. 60% for CXR (SE = 0.070, p < 0.01).ConclusionAccounting for patient body habitus as determined by either BMI, height, or weight did not improve the correlation between CXR accuracy and chest CT accuracy. CXR is significantly less accurate than CT even in patients with a low BMI. creator: Thomas F. Heston creator: John Y. Jiang uri: https://doi.org/10.7717/peerj.15090 license: https://creativecommons.org/publicdomain/zero/1.0/ rights: title: An in silico approach to the identification of diagnostic and prognostic markers in low-grade gliomas link: https://peerj.com/articles/15096 last-modified: 2023-03-16 description: Low-grade gliomas (LGG) are central nervous system Grade I tumors, and as they progress they are becoming one of the deadliest brain tumors. There is still great need for timely and accurate diagnosis and prognosis of LGG. Herein, we aimed to identify diagnostic and prognostic biomarkers associated with LGG, by employing diverse computational approaches. For this purpose, differential gene expression analysis on high-throughput transcriptomics data of LGG versus corresponding healthy brain tissue, derived from TCGA and GTEx, respectively, was performed. Weighted gene co-expression network analysis of the detected differentially expressed genes was carried out in order to identify modules of co-expressed genes significantly correlated with LGG clinical traits. The genes comprising these modules were further used to construct gene co-expression and protein-protein interaction networks. Based on the network analyses, we derived a consensus of eighteen hub genes, namely, CD74, CD86, CDC25A, CYBB, HLA-DMA, ITGB2, KIF11, KIFC1, LAPTM5, LMNB1, MKI67, NCKAP1L, NUSAP1, SLC7A7, TBXAS1, TOP2A, TYROBP, and WDFY4. All detected hub genes were up-regulated in LGG, and were also associated with unfavorable prognosis in LGG patients. The findings of this study could be applicable in the clinical setting for diagnosing and monitoring LGG. creator: Melih Özbek creator: Halil Ibrahim Toy creator: Yavuz Oktay creator: Gökhan Karakülah creator: Aslı Suner creator: Athanasia Pavlopoulou uri: https://doi.org/10.7717/peerj.15096 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Özbek et al. title: Marinated oven-grilled beef entrecôte meat from a bovine farm: Evaluation of resultant physicochemical and organoleptic attributes link: https://peerj.com/articles/15116 last-modified: 2023-03-16 description: Understanding the impact that combined action of marination and oven grill processes would have on such meat products as beef entrecôte is crucial from both consumer appeal and product development standpoints. Therefore, different marinated oven-grilled beef entrecôte meat specifically evaluating resultant physicochemical and organoleptic attributes were studied. The beef entrecôte meat was provided by a reputable local bovine farm/slaughter at Wroclaw, Poland. Physicochemical attributes involved antioxidant (2,2′-azinobis(3-ethylbenzothiaziline-6-sulfonate) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP)), (pH, thiobarbituric acid reactive substance (TBARS), cooking weight loss, L*a*b* color, and textural cutting force). Organoleptic attributes involved sensory (flavour, appearance, tenderness, taste) and texture (hardness, chewiness, gumminess, graininess, and greasiness) aspects. Different marination variants involved constituent 0.5%, 1%, and 1.5% quantities of cranberry pomace (CP), grape pomace (GP), and Baikal skullcap (BS), subsequently incorporated either African spice (AS) or industrial marinade/pickle (IM). Results showed pH, ABTS, DPPH, FRAP, TBARS, L*a*b* color, cooking weight loss, and textural cutting force, sensory and textural profile with varying range values. Concentration increases of either CP, GP, and or BS may not always go along with ABTS, DPPH, and FRAP values, given the observed decreasing or increasing fluctuations. As oven-grilling either increased or decreased the TBARS values alongside some color and textural cutting force trends, pH variations by difference seemed more apparent at samples involving GP, before CP, and then BS. The organoleptic attributes obtained differences and resemblances from both sensory and textural profile standpoints. Overall, oven-grilling promises to moderate both physicochemical and organoleptic range values of different marinated beef entrecôte meat samples in this study. creator: Charles Odilichukwu R. Okpala creator: Szymon Juchniewicz creator: Katarzyna Leicht creator: Małgorzata Korzeniowska creator: Raquel P. F. Guiné uri: https://doi.org/10.7717/peerj.15116 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Okpala et al. title: Health-related quality of life and associated factors after hip fracture. Results from a six-month prospective cohort study link: https://peerj.com/articles/14671 last-modified: 2023-03-15 description: BackgroundHip fractures are a major public health problem with increasing relevance in aging societies. They are associated with high mortality rates, morbidity, and loss of independence. The aim of the EMAAge study was to determine the impact of hip fractures on patient-reported health-related quality of life (HRQOL), and to identify potential risk factors for worse outcomes.MethodsEMAAge is a multicenter, prospective cohort study of patients who suffered a hip fracture. Patients or, if necessary, proxies were interviewed after initial treatment and after six months using standardized questionnaires including the EQ-5D-5L instrument, the Oxford Hip Score, the PHQ-4, the Short Nutritional Assessment Questionnaire, and items on patients living situation. Medical data on diagnoses, comorbidities, medications, and hospital care were derived from hospital information systems.ResultsA total of 326 patients were included. EQ-5D index values decreased from a mean of 0.70 at baseline to 0.63 at six months. The mean self-rated health on the EQ-VAS decreased from 69.9 to 59.4. Multivariable linear regression models revealed three relevant associated factors with the six-months EQ-5D index: symptoms of depression and anxiety, pre-fracture limitations in activities of daily living, and no referral to a rehabilitation facility had a negative impact. In addition, the six-months EQ-VAS was negatively associated with polypharmacy, living in a facility, and migration background.ConclusionsHip fractures have a substantial negative impact on patients HRQOL. Our results suggest that there are modifying factors that need further investigation including polypharmacy and migration background. Structured and timely rehabilitation seems to be a protective factor. creator: Johannes Deutschbein creator: Tobias Lindner creator: Martin Möckel creator: Mareen Pigorsch creator: Gabriela Gilles creator: Ulrich Stöckle creator: Ursula Müller-Werdan creator: Liane Schenk uri: https://doi.org/10.7717/peerj.14671 license: https://creativecommons.org/licenses/by/4.0/ rights: 2023 Deutschbein et al. title: The effect of a combined sprint training intervention on sprint force-velocity characteristics in junior Australian football players link: https://peerj.com/articles/14873 last-modified: 2023-03-15 description: BackgroundSprint performance in junior Australian football (AF) players has been shown to be a differentiating quality in ability level therefore developing sprint characteristics via sprint-specific training methods is an important aspect of their physical development. Assisted sprint training is one training method used to enhance sprint performance yet limited information exists on its effect on sprint force-velocity characteristics. Therefore, the main aim of this study was to determine the influence of a combined sprint training intervention using assisted and maximal sprint training methods on mechanical characteristics and sprint performance in junior Australian football players.MethodsUpon completing familiarization and pre-testing, twenty-two male junior Australian football (AF) players (age 14.4 ± 0.3 years, body mass 58.5 ± 10.0 kg, and height 1.74 ± 0.08 m) were divided into a combined sprint training (CST) group (n = 14), and a maximal sprint training (MST) group (n = 8) based on initial sprint performance over 20-meters. Sprint performance was assessed during maximal 20-meter sprint efforts via a radar gun (36 Hz), with velocity-time data used to derive force-velocity characteristics and split times. All subjects then completed a 7-week in-season training intervention consisting of maximal sprinting (MST & CST groups) and assisted sprinting (CST only), along with their usual football specific exercises.ResultsModerate to large pre-post within group effects (−0.65 ≤ ES ≥ 0.82. p ≤ 0.01) in the CST group for relative theoretical maximal force (F0) and power (Pmax) were reflected in improved sprint performance from 0–20 m, thereby creating a more force-oriented F-v profile. The MST group displayed statistically significant pre-post differences in sprint performance between 10–20 m only (ES = 0.18, p = 0.04). Moderate to high relative reliability was achieved across all sprint variables (ICC = 0.65–0.91), except for the force-velocity slope (SFV) and decrement in ratio of forces (DRF) which reported poor reliability (ICC = 0.41–0.44), while the CST group exceeded the pre-post minimal detectable change (MDC) in most sprint variables suggesting a ‘true change’ in performance across the intervention.ConclusionIt is concluded that implementing a short-term, combined sprint training intervention consisting of assisted and maximal sprint training methods may enhance sprint mechanical characteristics and sprint performance to 20-meters in junior AF players. creator: Dylan Shaun Hicks creator: Claire Drummond creator: Kym J. Williams creator: Roland van den Tillaar uri: https://doi.org/10.7717/peerj.14873 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Hicks et al. title: Minimal effects of ultraviolet light supplementation on egg production, egg and bone quality, and health during early lay of laying hens link: https://peerj.com/articles/14997 last-modified: 2023-03-15 description: Chicken vision is sensitive to ultraviolet (UV) light containing the UVA spectrum, while UVB plays a key role in the endogenous production of vitamin D3. However, commercially available light sources are typically deficient in the UV spectrum and thus may not adequately fulfill the lighting requirements of indoor-housed laying hens. We hypothesized that supplementary UVB light may improve egg production and egg quality, and bone health during early lay relative to UVA supplementation or standard control lighting. To investigate the effects of UV light supplementation, an experiment was conducted on 252 ISA Brown hens during 16 to 27 weeks of age. Birds were housed in eighteen pens (14 hens/pen) under three different light treatment groups each with six replications: (i) UVO: standard control lighting with LED white light, (ii) UVA: control lighting plus supplemental daylight with an avian bulb, and (iii) UVA/B: control lighting plus a supplemental full spectrum reptile bulb containing both UVA and UVB wavelengths. Hen-day egg production and egg quality, blood parameters including plasma Ca and P, and serum 25(OH)D3, and hen body weight and external health scoring were measured at different age points; while bone quality was assessed at the end of the experiment at 27 weeks. Data were analyzed in JMP® 16.0 using general linear mixed models with α level set at 0.05. Results showed that UVA and UVA/B supplemented birds reached sexual maturity (50% production) 3 and 1 day earlier, respectively, than control birds. There was a trend for UV lights to increase hen-day egg production (P = 0.06). Among egg quality traits, only eggshell reflectivity and yolk index were affected by UV lights (P = 0.02 and 0.01, respectively); however, most of the egg quality traits changed over age (all P < 0.01). Post-hoc tests showed higher serum 25(OH)D3 in the UVA/B group relative to control hens (P < 0.05); but there was no treatment effect on plasma Ca and P or on bone quality parameters (all P > 0.05). A significant interaction was observed between light treatment and age for the number of comb wounds (P = 0.0004), with the UV supplemented hens showing more comb wounds after 24 weeks. These results demonstrated that supplemental UVA/B light had minimal effects on egg production and egg quality, whereas, UVA/B exposure may increase vitamin D3 synthesis during the early laying period. The optimum duration of exposure and level of intensity needs to be determined to ensure these benefits. creator: Md Sohel Rana creator: Jonathon Clay creator: Prafulla Regmi creator: Dana L.M. Campbell uri: https://doi.org/10.7717/peerj.14997 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Rana et al. title: Metabolomics combined with clinical analysis explores metabolic changes and potential serum metabolite biomarkers of antineutrophil cytoplasmic antibody-associated vasculitis with renal impairment link: https://peerj.com/articles/15051 last-modified: 2023-03-15 description: BackgroundAntineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is an autoimmune systemic disease, and the majority of AAV patients have renal involvement presenting as rapid progressive glomerulonephritis (GN). Currently, the clinically available AAV markers are limited, and some of the newly reported markers are still in the nascent stage. The particular mechanism of the level changes of various markers and their association with the pathogenesis of AAV are not well defined. With the help of metabolomics analysis, this study aims to explore metabolic changes in AAV patients with renal involvement and lay the foundation for the discovery of novel biomarkers for AAV-related kidney damage.MethodsWe performed liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based on serum samples from patients with AAV (N = 33) and healthy controls (N = 33) in order to characterize the serum metabolic profiling. The principal component analysis (PCA) and orthogonal partial least-squares-discriminant analysis (OPLS-DA) were used to identify the differential metabolites. Least Absolute Shrinkage and Selection Operator (LASSO) and eXtreme Gradient Boosting (XGBoost) analysis were further conducted to identify the potential diagnostic biomarker. A receiver operating characteristic (ROC) curve analysis was conducted to evaluate the diagnostic performance of the identified potential biomarker.ResultsA total of 455 metabolites were detected by LC-MS analysis. PCA and OPLS-DA demonstrated a significant difference between AAV patients with renal involvement and healthy controls, and 135 differentially expressed metabolites were selected, with 121 upregulated and 14 downregulated. Ninety-two metabolic pathways were annotated and enriched based on the KEGG database. N-acetyl-L-leucine, Acetyl-DL-Valine, 5-hydroxyindole-3-acetic acid, and the combination of 1-methylhistidine and Asp-phe could accurately distinguish AAV patients with renal involvement from healthy controls. And 1-methylhistidine was found to be significantly associated with the progression and prognosis of AAV with renal impairment. Amino acid metabolism exhibits significant alternations in AAV with renal involvement.ConclusionThis study identified metabolomic differences between AAV patients with renal involvement and non-AAV individuals. Metabolites that could accurately distinguish patients with AAV renal impairment from healthy controls in this study, and metabolites that were significantly associated with disease progression and prognosis were screened out. Overall, this study provides information on changes in metabolites and metabolic pathways for future studies of AAV-related kidney damage and lays a foundation for the exploration of new biomarkers of AAV-related kidney damage. creator: Siyang Liu creator: Qing Xu creator: Yiru Wang creator: Yongman Lv creator: Qing quan Liu uri: https://doi.org/10.7717/peerj.15051 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Liu et al. title: Knowledge, attitude, and practices of front line health workers after receiving a COVID-19 vaccine: a cross-sectional study in Pakistan link: https://peerj.com/articles/14727 last-modified: 2023-03-14 description: BackgroundGlobally, there is an increased risk of COVID-19 infection among front-line health workers (FHW). This study aimed to evaluate the knowledge, attitude, and practices of FHW of Pakistan after receiving the COVID-19 vaccine.MethodsA population web-based survey on COVID-19 vaccine was conducted on 635 FHW in Pakistan between April 15, 2021, and July 15, 2021. The survey focused on four main sections consisting of socio-demographic data, knowledge, attitude, and practices after receiving the COVID-19 vaccine. The data was analyzed on SPSS. p < 0.05 was considered significant.ResultsOverall, 60% of FHW were nervous before getting vaccinated, with the leading reason to get vaccinated being their concern to protect themselves and their community (53.4%). A majority of FHW had fear about the unseen side effects of the COVID-19 vaccine (59.7%) used in Pakistan, with the most common side effect reported as soreness at the injection site (39%). It has been noted that almost all of the FHW observed preventive practices after getting vaccinated. The results showed that married respondents had favorable practices towards COVID-19 vaccines (B = 0.53, p < 0.01) (B, unstandardized regression coefficient). It was also found that more informational sources (B = 0.19, p < 0.01), higher knowledge of vaccination (B = 0.15, p < 0.001), and favorable attitude toward vaccine (B = 0.12, p < 0.001) significantly predicted favorable practices toward COVID-19 vaccination.ConclusionThe findings reflect that FHW, though they were worried about its side effects, have good knowledge and a positive attitude after getting the COVID-19 vaccine. This study is significant as the FHWs are a symbol for guidance, a reliable source of information, and an encouraging means of receiving COVID-19 vaccine for the general public. This study also reported that post-vaccination side effects were mild which will aid in reducing the vaccine hesitancy among the general Pakistani population. creator: Sadia Minhas creator: Aneequa Sajjad creator: Iram Manzoor creator: Atika Masood creator: Agha Suhail creator: Gul Muhammad Shaikh creator: Muhammad Kashif uri: https://doi.org/10.7717/peerj.14727 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Minhas et al. title: A tragedy of the commons case study: modeling the fishers king crab system in Southern Chile link: https://peerj.com/articles/14906 last-modified: 2023-03-14 description: Illegal fishing in small-scale fisheries is a contentious issue and resists a straightforward interpretation. Particularly, there is little knowledge regarding cooperative interactions between legal and illegal fishers and the potential effects on fisheries arising from these interactions. Taking the Chilean king crab (Lithodes santolla; common name centolla) fishery as a case study, our goal is twofold: (i) to model the effect of illegal-legal fishers’ interactions on the fishery and (ii) analyze how management and social behavior affect fishery’s outcomes. We framed the analysis of this problem within game theory combined with network theory to represent the architecture of competitive interactions. The fishers’ system was set to include registered (legal) fishers and unregistered (illegal) fishers. In the presence of unregistered fishers, legal fishers may decide to cooperate (ignoring the presence of illegal fishers) or defect, which involves becoming a “super fisher” and whitewashing the captures of illegal fishers for a gain. The utility of both players, standard fisher and super fisher depend on the strategy chosen by each of them, as well as on the presence of illegal fishers. The nodes of the network represent the legal fishers (both standard and super fishers) and the links between nodes indicate that these fishers compete for the resource, assumed to be finite and evenly distributed across space. The decision to change (or not) the adopted strategy is modeled considering that fishers are subjected to variable levels of temptation to whitewash the illegal capture and to social pressure to stop doing so. To represent the vital dynamics of the king crab, we propose a model that includes the Allee effect and a term accounting for the crab extraction. We found that the super fisher strategy leads to the decrease of the king crab population under a critical threshold as postulated in the tragedy of the commons hypothesis when there are: (i) high net extraction rates of the network composed of non-competing standard fishers, (ii) high values of the extent of the fishing season, and (iii) high density of illegal fishers. The results suggest that even in the presence of super fishers and illegal fishers, the choice of properly distributed fishing/closure cycles or setting an extraction limit per vessel can prevent the king crab population from falling below a critical threshold. This finding, although controversial, reflects the reality of this fishery that, for decades, has operated under a dynamic in which whitewashing and super fishers have become well established within the system. creator: Alan Zambrano creator: María F. Laguna creator: Marcelo N. Kuperman creator: Pedro Laterra creator: Jorge A. Monjeau creator: Laura Nahuelhual uri: https://doi.org/10.7717/peerj.14906 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Zambrano et al. title: Structure-based computational screening of 470 natural quercetin derivatives for identification of SARS-CoV-2 Mpro inhibitor link: https://peerj.com/articles/14915 last-modified: 2023-03-14 description: Coronavirus disease 2019 (COVID-19) is a global pandemic infecting the respiratory system through a notorious virus known as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Due to viral mutations and the risk of drug resistance, it is crucial to identify new molecules having potential prophylactic or therapeutic effect against SARS-CoV-2 infection. In the present study, we aimed to identify a potential inhibitor of SARS-CoV-2 through virtual screening of a compound library of 470 quercetin derivatives by targeting the main protease—Mpro (PDB ID: 6LU7). The study was carried out with computational techniques such as molecular docking simulation studies (MDSS), molecular dynamics (MD) simulations, and molecular mechanics generalized Born surface area (MMGBSA) techniques. Among the natural derivatives, compound 382 (PubChem CID 65604) showed the best binding affinity to Mpro (−11.1 kcal/mol). Compound 382 interacted with LYS5, TYR126, GLN127, LYS137, ASP289, PHE291, ARG131, SER139, GLU288, and GLU290 of the Mpro protein. The SARS-CoV-2 Mpro-382 complex showed acceptable stability during the 100 ns MD simulations. The SARS-CoV-2 Mpro-382 complex also showed an MM-GBSA binding free energy value of -54.0 kcal/mol. The binding affinity, stability, and free energy results for 382 and Mpro were better than those of the native ligand and the standard inhibitors ledipasvir and cobicistat. The conclusion of our study was that compound 382 has the potential to inhibit SARS-Cov-2 Mpro. However, further investigations such as in-vitro assays are recommended to confirm its in-silico potency. creator: Abd. Kakhar Umar creator: James H. Zothantluanga creator: Jittima Amie Luckanagul creator: Patanachai Limpikirati creator: Sriwidodo Sriwidodo uri: https://doi.org/10.7717/peerj.14915 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Kakhar Umar et al. title: Data-driven models for flood prediction in an ungauged karst wetland: Napahai wetland, Yunnan, China link: https://peerj.com/articles/14940 last-modified: 2023-03-14 description: Flood prediction for ungauged karst wetland is facing a great challenge. How to build a wetland hydrological model when there is a lack of basic hydrological data is the key to dealing with the above challenge. Napahai wetland is a typical ungauged karst wetland. In ungauged wetland/condition, this article used the wetland open water area (OWA) extracted from Landsat remote sensing images during 1987–2018 to characterize the hydrological characteristics of Napahai wetland. The local daily precipitation in the 1987–2018 rainy season (June–October) was used to set the variables. Based on the following hypothesis: in the rainy season, the OWA of the Napahai wetland rises when there is an increase in accumulated precipitation (AP), two data-driven models were established. The study took the area difference (AD) between two adjacent OWAs as the dependent variable, the accumulated precipitation (AP) within the acquisition time of two adjacent OWAs, and the corresponding time interval (TI) of the OWA as explanatory variables. Two data-driven models (a piecewise linear regression model and a decision tree model) were established to carry out flood forecasting simulations. The decision tree provided higher goodness of fit while the piecewise linear regression could offer a better interpretability between the variables which offset the decision tree. The results showed that: (1) the goodness of fit of the decision tree is higher than that of the piecewise linear regression model (2) the piecewise linear model has a better interpretation. When AP increased by 1 mm, the average AD increased by 2.41 ha; when TI exceeded 182 d and increased by 1 d, the average AD decreased to 3.66 ha. This article proposed an easy decision plan to help the local Napahai water managers forecast floods based on the results from the two models above. In addition, the modelling method proposed in this article, based on the idea of difference for non-equidistant time series, can be applied to karst wetland hydrological simulation problems with data acquisition difficulty. creator: Xiao Li creator: Jie Li uri: https://doi.org/10.7717/peerj.14940 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Li and Li title: A pilot study to develop assessment tools for Group A Streptococcus surveillance studies link: https://peerj.com/articles/14945 last-modified: 2023-03-14 description: IntroductionGroup A Streptococcus (GAS) causes pharyngitis (sore throat) and impetigo (skin sores) GAS pharyngitis triggers rheumatic fever (RF) with epidemiological evidence supporting that GAS impetigo may also trigger RF in Australian Aboriginal children. Understanding the concurrent burden of these superficial GAS infections is critical to RF prevention. This pilot study aimed to trial tools for concurrent surveillance of sore throats and skins sore for contemporary studies of RF pathogenesis including development of a sore throat checklist for Aboriginal families and pharynx photography.MethodsYarning circle conversations and semi-structured interviews were performed with Aboriginal caregivers and used to develop the language and composition of a sore throat checklist. The sore throat story checklist was combined with established methods of GAS pharyngitis and impetigo surveillance (examination, bacteriological culture, rapid antigen detection and serological tests) and new technologies (photography) and used for a pilot cross-sectional surveillance study of Aboriginal children attending their health clinic for a routine appointment. Feasibility, acceptability, and study costs were compiled.ResultsTen Aboriginal caregivers participated in the sore-throat yarning circles; a checklist was derived from predominant symptoms and their common descriptors. Over two days, 21 Aboriginal children were approached for the pilot surveillance study, of whom 17 were recruited; median age was 9 years [IQR 5.5–13.5], 65% were female. One child declined throat swabbing and three declined finger pricks; all other surveillance elements were completed by each child indicating high acceptability of surveillance assessments. Mean time for screening assessment was 19 minutes per child. Transport of clinical specimens enabled gold standard microbiological and serological testing for GAS. Retrospective examination of sore throat photography concorded with assessments performed on the day.ConclusionYarning circle conversations were effective in deriving culturally appropriate sore throat questionnaires for GAS pharyngitis surveillance. New and established tools were feasible, practical and acceptable to participants and enable surveillance to determine the burden of superficial GAS infections in communities at high risk of RF. Surveillance of GAS pharyngitis and impetgio in remote Australia informs primary RF prevention with potential global translation. creator: Janessa Pickering creator: Claudia Sampson creator: Marianne Mullane creator: Meru Sheel creator: Dylan D. Barth creator: Mary Lane creator: Roz Walker creator: David Atkinson creator: Jonathan R. Carapetis creator: Asha C. Bowen uri: https://doi.org/10.7717/peerj.14945 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Pickering et al. title: Response of dung beetle diversity to remediation of soil ecosystems in the Ecuadorian Amazon link: https://peerj.com/articles/14975 last-modified: 2023-03-14 description: BackgroundEfforts to alleviate the negative effects of oil spills in the Ecuadorian Amazon include remediation activities such as cleaning, reshaping, and revegetation of polluted areas. However, studies of the diversity of biological communities in these hydrocarbon-degraded ecosystems have never been carried out. Here, we evaluated the diversity of dung beetles on remediated soil ecosystems (Agricultural Soils and Sensitive Ecosystems) and on non-contaminated soils (Natural Forests and Palm Plantations).MethodologyThe study was conducted in Sucumbíos and Orellana provinces, in the Ecuadorian Amazon at four sampling sites per ecosystem type (a total of 16 sites). At each sampling site, six pitfall traps remained active for 120 consecutive h per month for 1 year.ResultsWe collected 37 species and 7,506 individuals of dung beetles. We observed significant differences in mean species abundance, richness, and diversity between non-contaminated soil ecosystems and remediated soil ecosystems, with Natural Forests presenting the highest values, and Agricultural Soils the lowest values. Regarding sampling month, we also found significant differences among ecosystems, which were also higher in Natural Forests.DiscussionThe results suggest that hydrocarbon-degraded ecosystems tend to conserve lower beetle diversity one year after remediation highlighting the importance of Natural Forests for the conservation of tropical biodiversity. Therefore, dung beetle diversity could be used for future landscape management of these hydrocarbon-degraded ecosystems. creator: Wilmer E. Pozo-Rivera creator: Carlos Quiloango-Chimarro creator: Xavier Paredes creator: Mario Landívar creator: Carlos Chiriboga creator: Daniel Hidalgo creator: Karina García creator: Jaime Villacís uri: https://doi.org/10.7717/peerj.14975 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Pozo-Rivera et al. title: Wearing N95 masks decreases the odor discrimination ability of healthcare workers: a self-controlled before-after study link: https://peerj.com/articles/14979 last-modified: 2023-03-14 description: ObjectiveDuring the coronavirus disease 2019 (COVID-19) pandemic, the N95 mask is an essential piece of protective equipment for healthcare workers. However, the N95 mask may inhibit air exchange and odor penetration. Our study aimed to determine whether the use of N95 masks affects the odor discrimination ability of healthcare workers.MethodsIn our study, all the participants were asked to complete three olfactory tests. Each test involved 12 different odors. The participants completed the test while wearing an N95 mask, a surgical mask, and no mask. The score for each olfactory test was documented.ResultsThe olfactory test score was significantly lower when the participants wore N95 masks than when they did not wear a mask (7 vs. 10, p < 0.01). The score was also lower when the participants wore N95 masks than surgical masks (7 vs. 8, p < 0.01).ConclusionWearing N95 masks decreases the odor discrimination ability of healthcare workers. Therefore, we suggest that healthcare workers seek other clues when diagnosing disease with a characteristic odor. creator: Guanguan Luo creator: Xingnan Zou creator: Xianlong Zhou creator: Jiaohong Gan creator: Cheng Jiang creator: Zhigang Zhao creator: Yan Zhao uri: https://doi.org/10.7717/peerj.14979 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Luo et al. title: Transition of an estuarine benthic meiofauna assemblage 1.7 and 2.8 years after a mining disaster link: https://peerj.com/articles/14992 last-modified: 2023-03-14 description: BackgroundEstuaries are transitional coastal ecosystems that are threatened by multiple sources of human pollution. In 2015, mining tailings from an upstream dam failure caused massive metal contamination that impacted benthic assemblages on the Brazilian Rio Doce estuary.MethodsIn this study, we investigate and compare meiofaunal assemblages with eDNA metabarcoding 1.7 years (2017) and 2.8 years (2018) after the initial contamination by mine tailings in order to evaluate the continued impact of sediment mine tailing contaminants on the structure of benthic assemblages after the disaster.ResultsThe community was dominated by Arthropoda and Nematoda 1.7 yr after the impacts (42 and 29% of meiofaunal sequence reads, respectively) but after 2.8 years Arthropoda (64.8% of meiofaunal sequence reads) and Rotifera (11.8%) were the most common taxa. This continued impact on meiofaunal assemblage revealed a lower phylogenetic diversity (7.8-fold) in 2018, despite overall decrease in metal concentration (Al, Ba, Cr, As, Fe, Zn, Mn, Pb, Cd, Co) in sediments. Our data suggests that differences in benthic assemblages and loss of diversity may be influenced by contaminants in sediments of this estuary, and indicate that broad eDNA assessments are greatly useful to understand the full range of biodiversity changes in dynamic estuarine ecosystems. creator: Gabriel Coppo creator: Fabiano S. Pais creator: Tiago O. Ferreira creator: Ken M. Halanych creator: Kyle Donnelly creator: Ana Carolina Mazzuco creator: Angelo F. Bernardino uri: https://doi.org/10.7717/peerj.14992 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Coppo et al. title: Cloning, characterization, and expression analysis of the CHITINASE gene family in Helice tientsinensis link: https://peerj.com/articles/15045 last-modified: 2023-03-14 description: Chitinase is a kind of glycoside hydrolase which is widely distributed in nature and encoded by multiple genes to catalyze the decomposition of chitin, which plays an important role in the molting and pathogen defense of crustaceans. However, the research on chitinase in crustaceans is mainly focused on a few species with economic value. In this study, full-length cDNA sequences of the HtCHT1, HtCHT3 and HtCHT4 genes were cloned from the mudflat crab Helice tientsinensis by RACE, and the sequences were analyzed. The results showed that the full-length 2,229 bp of HtCHT1 gene encoded 627 amino acids, while the full-length 2,191 bp of HtCHT3 gene produced 489 amino acids, and the full-length 3,312 bp of HtCHT4 gene encoded 664 amino acids. Bioinformatics analysis showed that all the obtained chitinase proteins had the glycosyl hydrolase family 18 (GH18) catalytic domain and chitin-binding domain (ChtBD2), furthermore, HtCHT1 and HtCHT4 proteins had signal peptide domains at N-terminal. Phylogenetic analysis showed that different types of chitinase were clustered, and HtCHTs were closely related to chitinases in the Eriocheir sinensis. Expression profile analysis showed that the HtCHT1, HtCHT3 and HtCHT4 were significantly expressed in hepatopancreas. Furthermore, the expression of three genes was significantly up-regulated in hepatopancreas after the Vibrio parahaemolyticus challenge. These results suggested that HtCHT1, HtCHT3 and HtCHT4 were belonged to the CHITINASE gene family in H. tientsinensis and were potentially involved in the antibacterial immune response. This study provides essential information for further research of chitinase in H. tientsinensis and even crustaceans. creator: Lulu Chen creator: Yuyan Hua creator: Wenxuan Ji creator: Jiayu Wang creator: Hua Zhao creator: Zhengfei Wang uri: https://doi.org/10.7717/peerj.15045 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Chen et al. title: An efficient transformation method for tannin-containing sorghum link: https://peerj.com/articles/15066 last-modified: 2023-03-14 description: BackgroundTannins are the main bottlenecks restricting the transformation efficiency of plants. Hongyingzi is a special tannin-containing sorghum cultivar used in brewing.MethodsIn this study, a highly efficient microprojectile transformation system for tannin-containing sorghum was successfully exploited using immature embryos (IEs) of Hongyingzi as explants.ResultsHongyingzi presented two types of calli. Type II calli were found to be the most suitable and effective explants for transformation. After optimization of the geneticin (G418) concentration and tissue culture medium, an average transformation frequency of 27% was achieved. Molecular analyzis showed that all transgenic plants were positive and showed transgenes expression. The inheritance analyzis confirmed that the transgenes could be inherited into the next generation. Thus, we successfully established an efficient transformation system for tannin-containing sorghum and demonstrated the possibility of breaking the restriction imposed by tannins in plants. creator: Yuan Liang creator: Xuehui Yan creator: Jingyi Xu creator: Yanlong Liu creator: Ke Xie creator: Jieqin Li creator: Qiuwen Zhan uri: https://doi.org/10.7717/peerj.15066 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Liang et al. title: Preliminary study on Bioassay of Capparis spinosa L. seed extract and seed germination link: https://peerj.com/articles/15082 last-modified: 2023-03-14 description: The present study explored the germination inhibitors present in the seeds of Capparis spinosa L., a plant species that is known for its ecological significance in preventing wind erosion and fixing sand in desertified areas. Additionally, its roots, leaves, and fruits possess medicinal properties, and are used to treat a range of ailments such as rheumatism, tumors, and diabetes. However, the plant’s low germination rate under natural conditions is a major limitation. We aimed to improve the germination of C. spinosa seeds by investigating the effects of various infusions of caper seeds on the germination and seedling growth of Chinese cabbage seeds. A range of chemical reagents, hormonal immersions, and sand storage treatments were used to determine the differences in the germination rate of C. spinosa seeds. Our results revealed that among the various water extract concentrations tested, 100% water extract exhibited the strongest inhibitory effect on the germination and growth of the cabbage seeds, with a germination rate of (70.00 ± 0.09)%. Furthermore, the inhibitory effects on the germination and growth of cabbage seeds were found to be strongest when treated with the extract solution 1, yielding a germination rate of (83.33 ± 0.02)%. Notably, the leaves of Chinese cabbage seedlings turned yellow-green and yellow after treatment with the extract solution. These findings highlight the potential inhibitory effects of C. spinosa seed extracts on seed germination and growth and suggest that further research is needed to better understand the underlying mechanisms. The results of the germination experiment with methanol extract showed a sharp decline in the germination rate of Chinese cabbage seeds treated with 50% methanol extract, to (4.67 ± 0.02)%. These findings indicate the presence of germination-inhibiting substances in caper seeds. The highest germination potential was observed when the caper seeds were soaked in 30% PEG, reaching 35.00%. The highest germination rate, 19.33%, was observed when the seeds were soaked in 250 mg/L GA3 and 25 mmol/L NaCl. These results suggest that the germination inhibitor present in caper seeds affects the germination of cabbage seeds as well. The highest germination rate was achieved when the seeds were soaked with gibberellin. It is hoped that the research on the germination-inhibiting substances in caper seeds will provide a scientific foundation for improving and refining the artificial propagation and cultivation methods of this species. creator: Min Wang creator: Xiaolu Yuan creator: Liping Xu uri: https://doi.org/10.7717/peerj.15082 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Wang et al. title: Cloning and comparative modeling identifies a highly stress tolerant Cu/Zn cytosolic super oxide dismutase 2 from a drought tolerant maize inbred line link: https://peerj.com/articles/14845 last-modified: 2023-03-13 description: Plants have a complex system of stress response that deals with different types of stresses. Maize (Zea mays L.), one of the most important crops grown throughout the world, across a range of agro-ecological environments, employs complex mechanisms of gene regulation in response to drought stress. HKI 335 is a tropical maize inbred line showing remarkable adaptation to drought stress. Abiotic stresses, like drought, trigger the production of reactive oxygen species (ROS) due to the incomplete reduction or excitation of molecular oxygen, eventually leading to cell damage. Superoxide dismutase (SOD, EC 1.15.1.1) is a metalloenzyme that acts as the first line of defense against ROS. We cloned the Sod2 gene from HKI 335 inbred line and analyzed its protein through detailed in silico characterization. Our comparative modeling revealed that at the level of tertiary structure, the HKI 335 SOD2 protein is highly similar to Potentilla atrosanguinea SOD2, which had been previously identified as highly thermostable SOD that can tolerate autoclaving as well as sub-zero temperatures. We performed phylogenetic analysis, estimated physicochemical properties, post-translational modifications, protein-protein interactions, and domain composition of this SOD2. The phylogenetic analysis showed that orthologous sequences of SOD from different species were clustered into two clusters. Secondary structure prediction indicates that SOD2 is a soluble protein and no transmembrane domains have been found. Most of the beta sheets have RSA value greater than 2. The Ramachandran plot from PDBsum revealed that most of the residues fall in the highly favored region. It was estimated that the value of the instability index was less than 40, the value of the aliphatic index was extremely high and the GRAVY value lies between −2 and +2. We could identify only one phosphorylation site, located at position 20 with a score of 0.692. Overall, the unique stress-tolerant properties of the HKI 335 SOD2, may be one of the reasons contributing to the high drought tolerance trait exhibited by HKI 335 maize inbred line. Further research may reveal more insights into the drought adaptation mechanism in maize and the eventual deployment of the trait in maize hybrids. creator: Anuradha Gautam creator: Fatima Nazish Khan creator: Surabhi Priya creator: Krishan Kumar creator: Shivani Sharda creator: Tanushri Kaul creator: Ishwar Singh creator: Sapna Langyan creator: Pranjal Yadava uri: https://doi.org/10.7717/peerj.14845 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Gautam et al. title: Thermal requirements, fertility life table and biological parameters of Cleruchoides noackae (Hymenoptera: Mymaridae) at different temperatures link: https://peerj.com/articles/14911 last-modified: 2023-03-13 description: Cleruchoides noackae Lin & Huber (Hymenoptera: Mymaridae) was imported to Brazil in 2012, to manage the exotic pest Thaumastocoris peregrinus Carpintero & Dellapé (Hemiptera: Thaumastocoridae), which has been damaging eucalyptus plantations. Knowledge of the thermal requirements and the fertility life table of C. noackae is important to improve mass rearing methods for this parasitoid and the effectiveness of its release to manage T. peregrinus. The objective was to evaluate the development period, thermal requirements and the fertility life table of C. noackae at different temperatures. The egg-adult period of this parasitoid varied from 43 to 14 days at 15 °C and 30 °C, respectively. The emergence of C. noackae adults was higher at 15 °C, 18 °C, 21 °C and 24 °C than at 30 °C. Female and male C. noackae need 226.75 and 230.41 degree-days and temperatures higher than 10.06 °C and 9.90 °C, respectively, to complete egg-adult development. The number of parasitized eggs per C. noackae female was higher at 21 °C, 24 °C and 27 °C, with 5.82, 7.73 and 5.50 eggs, respectively, than at 30 °C (0.45). Cleruchoides noackae longevity was greater at 15 °C, 21 °C and 24 °C. The net reproductive rate of the parasitoid was higher at 21 °C and 24 °C than at 30 °C, 3.05, 4.70 and 0.16, respectively. The finite rate of increase of C. noackae was greater at 21 °C, 24 °C and 27 °C, than at 30 °C and the intrinsic rate of increase was negative at 30 °C, −0.100. The temperatures 21 °C and 24 °C and from 18 °C to 27 °C are the most adequate for the reproduction and population increase of C. noackae parasitizing eggs of T. peregrinus, respectively. creator: Luciane Katarine Becchi creator: Leonardo Rodrigues Barbosa creator: José Eduardo Serrão creator: José Cola Zanuncio creator: Marcus Vinicius Sampaio creator: Maurício Magalhães Domingues creator: Carlos Frederico Wilcken uri: https://doi.org/10.7717/peerj.14911 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Katarine Becchi et al. title: Methods of performance analysis in women’s Australian football: a scoping review link: https://peerj.com/articles/14946 last-modified: 2023-03-13 description: BackgroundThe first women’s Australian football (AF) professional competition was established in 2017, resulting in advancement in performance analysis capabilities within the sport. Given the specific constraints of women’s AF, it is currently unclear what match-play performance analysis methods and techniques are implemented. Therefore, the aim of this scoping review was to describe and critically appraise physical, technical, and tactical performance analysis methods that have been employed in women’s and girls’ AF match-play.MethodologyA systematic search was conducted on the 27th of June 2022 through five databases. Eligibility criteria were derived from the PCC framework with the population (P) of women and girls AF players, of any level of play; concepts (C) of interest which were measures, data, and methods related to the sport’s physical, technical, and tactical performance; and the context (C) of methods that analysed any match-play performance. A narrative synthesis was conducted using extracted study characteristic data such as sample size, population, time period, collection standards, evaluation metrics for results, and application of thematic categorisations of previous sports performance reviews. Critical appraisal of eligible studies’ methodologies was conducted to investigate research quality and identify methodological issues.ResultsFrom 183 studies screened, twelve eligible studies were included, which examined match-play through physical (9/12, 75%), technical (4/12, 33%), and tactical analysis (2/12, 17%). Running demands and game actions analysis were the most researched in senior women’s AF. Research into junior girls’ AF match-play performance has not been investigated. No research has been conducted on non-running physical demands, contact demands, acceleration, and tactical aspects of women’s AF. All studies utilised either inferential statistics or basic predictive models. Critical appraisal deemed most studies as low risk of bias (11/12, 92%), with the remaining study having satisfactory risk.ConclusionsFuture research utilising increased longitudinal and greater contextual data is needed to combat the prominent issue of data representativeness to better characterise performance within women’s and girls’ AF. Additionally, research involving junior and sub-elite AF players across the talent pathways is important to conduct, as it provides greater context and insight regarding development to support the evolving elite women’s AF competition. Women’s AF has been constrained by its resource environment. As such, suggestions are provided for better utilisation of existing data, as well as for the creation of new data for appropriate future research. Greater data generation enables the use of detailed machine learning predictions, neural networks, and network analysis to better represent the intertwined nature of match-play performance from technical, physical, and tactical data. creator: Braedan R. van der Vegt creator: Adrian Gepp creator: Justin W.L. Keogh creator: Jessica B. Farley uri: https://doi.org/10.7717/peerj.14946 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 van der Vegt et al. title: Sodicity stress differently influences physiological traits and anti-oxidant enzymes in pear and peach cultivars link: https://peerj.com/articles/14947 last-modified: 2023-03-13 description: BackgroundThe growth and physiological responses to sodicity stress of pear and peach are poorly understood. Insights into how sodicity stress alters tree physiology remain vital to developing salt tolerant scion and rootstock cultivars.MethodsThe effects of sodicity stress (soil pHs ~8.8) on tree growth and physiological traits of field grown trees of pear cultivars Punjab Beauty and Patharnakh, and peach cultivars Partap and Shan-e-Punjab were recorded using standard procedures. Sodicity-induced changes in oxidative stressors, proline, anti-oxidant enzymes and leaf ions were measured to draw inferences.ResultsSodicity-induced reductions in vegetative growth were particularly marked in Patharnakh pear and Partap peach compared with other cultivars. Although sodicity stress triggered a significant increase in leaf malondialdehyde (MDA) and hydrogen peroxide (H2O2), their levels relative to controls were much higher in peach than in pear; reflecting that peach suffered from greater oxidative stress. Interestingly, MDA and H2O2 levels did not seem to be deleterious enough to trigger proline-induced osmotic adjustment in pears. The activities of anti-oxidant enzymes strongly varied with the cultivar; specifically, the sodicity-induced increases in CAT and SOD activities were much higher in Punjab Beauty pear and Shan-e-Punjab peach. Principal Component Analysis revealed an explicit convergence between CAT and SOD activities in Punjab Beauty and Shan-e-Punjab cultivars in response to sodicity-induced oxidative stress. Correlation analysis revealed that leaf Na+ strongly inhibited tree growth in peach than in pear. Leaf K+ and proline were found to be the major osmolytes in sodicity-stressed pear and peach cultivars, respectively.ConclusionsWe have for the first time studied the effects of sodicity stress on important tree growth and physiological traits of commercially important pear and peach cultivars. Our findings revealed a marked suppressive effect of sodicity stress on tree growth in peach than in pear. The sodicity-induced upticks in leaf malondialdehyde, hydrogen peroxide and Na+ seemed to induce proline-mediated osmotic adjustment in peach but not in pear. The overall better sodicity tolerance in pear compared to peach was ascribed to increased activities of anti-oxidant enzymes catalase and superoxide dismutase enzymes together with restricted Na+ uptake and better leaf K+ levels. Further investigations are needed to elucidate the effects of sodicity stress on genetic and transcriptional changes, and on fruit yield and quality. creator: Anshuman Singh creator: Ashwani Kumar creator: Parbodh Chander Sharma creator: Raj Kumar creator: Rajender Kumar Yadav uri: https://doi.org/10.7717/peerj.14947 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Singh et al. title: Relationship between ferroptosis and mitophagy in cardiac ischemia reperfusion injury: a mini-review link: https://peerj.com/articles/14952 last-modified: 2023-03-13 description: Cardiovascular diseases (CVD), with high morbidity and mortality, seriously affect people’s life and social development. Clinically, reperfusion therapy is typically used to treat ischemic cardiomyopathy, such as severe coronary heart disease and acute myocardial infarction. However, reperfusion therapy can lead to myocardial ischemia reperfusion injury (MIRI), which can affect the prognosis of patients. Studying the mechanisms of MIRI can help us improve the treatment of MIRI. The pathological process of MIRI involves many mechanisms such as ferroptosis and mitophagy. Ferroptosis can exacerbate MIRI, and regulation of mitophagy can alleviate MIRI. Both ferroptosis and mitophagy are closely related to ROS, but there is no clear understanding of the relationship between ferroptosis and mitophagy. In this review, we analyzed the relationship between ferroptosis and mitophagy according to the role of mTOR, NLPR3 and HIF. In addition, simultaneous regulation of mitophagy and ferroptosis may be superior to single therapy for MIRI. We summarized potential drugs that can regulate mitophagy and/or ferroptosis, hoping to provide reference for the development of drugs and methods for MIRI treatment. creator: Cuihua Liu creator: Zunjiang Li creator: Botao Li creator: Wei Liu creator: Shizhong Zhang creator: Kuncheng Qiu creator: Wei Zhu uri: https://doi.org/10.7717/peerj.14952 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Liu et al. title: Brain mechanism of unfamiliar and familiar voice processing: an activation likelihood estimation meta-analysis link: https://peerj.com/articles/14976 last-modified: 2023-03-13 description: Interpersonal communication through vocal information is very important for human society. During verbal interactions, our vocal cord vibrations convey important information regarding voice identity, which allows us to decide how to respond to speakers (e.g., neither greeting a stranger too warmly or speaking too coldly to a friend). Numerous neural studies have shown that identifying familiar and unfamiliar voices may rely on different neural bases. However, the mechanism underlying voice identification of individuals of varying familiarity has not been determined due to vague definitions, confusion of terms, and differences in task design. To address this issue, the present study first categorized three kinds of voice identity processing (perception, recognition and identification) from speakers with different degrees of familiarity. We defined voice identity perception as passively listening to a voice or determining if the voice was human, voice identity recognition as determining if the sound heard was acoustically familiar, and voice identity identification as ascertaining whether a voice is associated with a name or face. Of these, voice identity perception involves processing unfamiliar voices, and voice identity recognition and identification involves processing familiar voices. According to these three definitions, we performed activation likelihood estimation (ALE) on 32 studies and revealed different brain mechanisms underlying processing of unfamiliar and familiar voice identities. The results were as follows: (1) familiar voice recognition/identification was supported by a network involving most regions in the temporal lobe, some regions in the frontal lobe, subcortical structures and regions around the marginal lobes; (2) the bilateral superior temporal gyrus was recruited for voice identity perception of an unfamiliar voice; (3) voice identity recognition/identification of familiar voices was more likely to activate the right frontal lobe than voice identity perception of unfamiliar voices, while voice identity perception of an unfamiliar voice was more likely to activate the bilateral temporal lobe and left frontal lobe; and (4) the bilateral superior temporal gyrus served as a shared neural basis of unfamiliar voice identity perception and familiar voice identity recognition/identification. In general, the results of the current study address gaps in the literature, provide clear definitions of concepts, and indicate brain mechanisms for subsequent investigations. creator: YuXiang Sun creator: Lili Ming creator: Jiamin Sun creator: FeiFei Guo creator: Qiufeng Li creator: Xueping Hu uri: https://doi.org/10.7717/peerj.14976 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Sun et al. title: The risks of low hemoglobin deferral in a large retrospective cohort of plasmapheresis donors and the influence factors of return for a subsequent donation in China link: https://peerj.com/articles/14999 last-modified: 2023-03-13 description: BackgroundAccording to the Technical Operation Procedures for Plasmapheresis Collection Station (2019) in China, plasmapheresis donors with low hemoglobin (Hb) levels (men <12.0 g/dL; women <11.0 g/dL) were deferred for at least 2 weeks. The purpose of this retrospective study was to survey the demographic characteristics of plasmapheresis donors with low Hb deferral (LHD) and identify at-risk LHD donors, so as to enhance donor safety and improve donation service management.MethodsFrom 2018 to 2020, a multi-center study involving plasmapheresis donors from 18 plasmapheresis centers in three provinces (Sichuan, Yunnan and Hunan) of China was conducted. Donor demographics (age, sex) and donation information (date of donation, first-time donors vs. repeat donors, the number of lifetime donations, the number of donations in the last 12 months, and whether the LHD donor returned for a subsequent donation) were collected. The Cochran-Mantel-Haenszel method was used to explore the risk factors for LHD while adjusting for the different provinces. Logistic regression analysis was used to investigate the factors influencing the return for a subsequent donation after LHD.ResultsA total of 497,039 plasmapheresis donors were included. Female donors’ LHD rate was 0.15% on average, while male donors’ LHD rate was 0.01%. Female donors aged 41–50 years old (OR: 2.276, 95% CI [1.333–3.887], p = 0.002) were more likely to experience LHD temporarily than those aged 18–30 years old. For female donors, compared with donations in the winter, they had a higher risk for LHD in the summer (OR: 2.217, 95% CI [1.670–2.943], p < 0.001), spring (OR: 2.402, 95% CI [1.806–3.196], p < 0.001), and fall (OR: 2.002, 95% CI [1.500–2.673], p < 0.001). Among the LHD donors, those who had donated more frequently in the past were more likely to return for a subsequent donation (p = 0.012).ConclusionsFemale donors were at a higher risk of LHD, particularly between the ages of 41 and 50. A clear seasonal pattern in the rate of LHD was observed. In the winter, the risk of LHD was the lowest; thus, it was advised to recruit plasmapheresis donors throughout the winter and to make the required adjustments for recruitment measures during other seasons. The number of previous donations was correlated with the return rate after LHD. Our observations could have practical implications for plasmapheresis donor management. creator: Guanglin Xiao creator: Demei Dong creator: Ya Wang creator: Changqing Li creator: Gong tian Huang creator: Hui Yang creator: Jing Huang creator: Fei Chen uri: https://doi.org/10.7717/peerj.14999 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Xiao et al. title: Characterization of viral pathogens associated with symptomatic upper respiratory tract infection in adults during a low COVID-19 transmission period link: https://peerj.com/articles/15008 last-modified: 2023-03-13 description: BackgroundThe epidemiology of respiratory tract infections (RTI) has dramatically changed over the course of the COVID-19 pandemic. A major effort in the clinical management of RTI has been directed toward diagnosing COVID-19, while the causes of other, common community RTI often remain enigmatic. To shed light on the etiological causes of RTI during a low COVID-19 transmission period in 2021, we did a pilot study using molecular testing for virologic causes of upper RTI among adults with respiratory symptoms from Almaty, Kazakhstan.MethodsAdults presenting at two public hospitals with respiratory symptoms were screened using SARS-CoV-2 PCR on nasopharyngeal swabs. A subset of RTI+, COVID-19-negative adults (n = 50) was then tested for the presence of common RTI viruses and influenza A virus (IAV). Next generation virome sequencing was used to further characterize the PCR-detected RTI pathogens.ResultsOf 1,812 symptomatic adults, 21 (1.2%) tested SARS-CoV-2-positive. Within the COVID-19 negative outpatient subset, 33/50 subjects (66%) had a positive PCR result for a common community RTI virus, consisting of human parainfluenza virus 3-4 (hPIV 3-4) in 25/50 (50%), rhinovirus (hRV) in 2 (4%), hPIV4-hRV co-infection in four (8%) and adenovirus or the OCR43/HKU-1 coronavirus in two (4%) cases; no IAV was detected. Virome sequencing allowed to reconstruct sequences of most PCR-identified rhinoviruses and hPIV-3/human respirovirus-3.ConclusionsCOVID-19 was cause to a low proportion of symptomatic RTI among adults. Among COVID-negative participants, symptomatic RTI was predominantly associated with hPIV and hRV. Therefore, respiratory viruses other than SARS-CoV-2 should be considered in the clinical management and prevention of adult RTI in the post-pandemic era. creator: Nurlan Sandybayev creator: Vyacheslav Beloussov creator: Vitaliy Strochkov creator: Maxim Solomadin creator: Joanna Granica creator: Sergey Yegorov uri: https://doi.org/10.7717/peerj.15008 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Sandybayev et al. title: Evaluation of DNA metabarcoding for identifying fish eggs: a case study on the West Florida Shelf link: https://peerj.com/articles/15016 last-modified: 2023-03-13 description: A critical factor in fisheries management is the protection of spawning sites for ecologically and economically important fish species. DNA barcoding (i.e., amplification and sequencing of the mitochondrial cytochrome c oxidase I (COI) gene) of fish eggs has emerged as a powerful technique for identifying spawning sites. However, DNA barcoding of individual fish eggs is time-consuming and expensive. In an attempt to reduce costs and effort for long-term fisheries monitoring programs, here we used DNA metabarcoding, in which DNA is extracted and amplified from a composited sample containing all the fish eggs collected at a given site, to identify fish eggs from 49 stations on the West Florida Shelf. A total of 37 taxa were recovered from 4,719 fish eggs. Egg distributions on the West Florida Shelf corresponded with the known habitat types occupied by these taxa, which included burrower, coastal pelagic, epipelagic, mesopelagic, demersal, deep demersal, commensal, and reef-associated taxa. Metabarcoding of fish eggs was faster and far less expensive than barcoding individual eggs; however, this method cannot provide absolute taxon proportions due to variable copy numbers of mitochondrial DNA in different taxa, different numbers of cells within eggs depending on developmental stage, and PCR amplification biases. In addition, some samples yielded sequences from more taxa than the number of eggs present, demonstrating the presence of contaminating DNA and requiring the application of a threshold proportion of sequences required for counting a taxon as present. Finally, we review the advantages and disadvantages of using metabarcoding vs. individual fish egg barcoding for long-term monitoring programs. creator: Mya Breitbart creator: Makenzie Kerr creator: Michael J. Schram creator: Ian Williams creator: Grace Koziol creator: Ernst Peebles creator: Christopher D. Stallings uri: https://doi.org/10.7717/peerj.15016 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Breitbart et al. title: Dihydrotanshinone I inhibits hepatocellular carcinoma cells proliferation through DNA damage and EGFR pathway link: https://peerj.com/articles/15022 last-modified: 2023-03-13 description: BackgroundThe incidence and mortality of hepatocellular carcinoma (HCC) are globally on the rise. Dihydrotanshinone I, a natural product isolated from Salvia miltiorrhiza Bunge, has attracted extensive attention in recent years for its anti-tumour proliferation efficiency.MethodsCell proliferations in hepatoma cells (Huh-7 and HepG2) were evaluated by MTT and colony formation assays. Immunofluorescence (IF) of 53BP1 and flow cytometry analysis were performed to detect DNA damage and cell apoptosis. Furthermore, network pharmacological analysis was applied to explore the potential therapeutic targets and pathway of dihydrotanshinone I.ResultsThe results showed that dihydrotanshinone I effectively inhibited the proliferation of Huh-7 and HepG2 cells. Moreover, dihydrotanshinone I dose-dependently induced DNA-damage and apoptosis in vitro. Network pharmacological analysis and molecular simulation results indicated that EGFR might be a potential therapeutic target of dihydrotanshinone I in HCC. Collectively, our findings suggested that dihydrotanshinone I is a novel candidate therapeutic agent for HCC treatment. creator: Linjun Wang creator: Xiangwei Xu creator: Dexing Chen creator: Chenghang Li uri: https://doi.org/10.7717/peerj.15022 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Wang et al. title: Value of lymph node ratio as a prognostic factor of recurrence in medullary thyroid cancer link: https://peerj.com/articles/15025 last-modified: 2023-03-13 description: Background and ObjectivesThe purpose of this study is to evaluate the relationship between lymph node status (the number of resected lymph nodes; the number of metastatic lymph nodes, LNM, and lymph node ratio, LNR) and biochemical recurrence, disease-free survival (DFS), as well as overall survival (OS) in medullary thyroid carcinoma (MTC).MethodsThis study enrolled MTC patients at Tianjin Medical University Cancer Institute and Hospital between 2011 and 2019. We used Logistic regression analysis, Cox regression models and Kaplan-Meier test to identify risk factors influencing biochemical recurrence, DFS, and OS.ResultsWe identified 160 patients who satisfied the inclusion criteria from 2011 to 2019. We used ROC analysis to define the cut-off value of LNR with 0.24. Multifocality, preoperative calcitonin levels, pathologic N stage, resected lymph nodes, LNM, LNR, and the American Joint Committee on Cancer (AJCC) clinical stage were significant (P < 0.05) prognostic factors influencing biochemical cure. In univariable analyses, gross extrathyroidal extension, preoperative calcitonin levels, pathologic T classification, pathologic N stage, resected lymph nodes, LNM, LNR, AJCC clinical stage, and biochemical cure were significant (P < 0.05) factors of DFS. When the multivariable analysis was performed, LNR was identified as predictor of DFS (HR = 4.818, 95% CI [1.270–18.276]). Univariable Cox regression models reflected that tumor size, pathologic N stage, and LNR were predictor of OS. Furthermore, multivariable analysis manifested that LNR was predictor of OS (HR = 10.061, 95% CI [1.222–82.841]).ConclusionsThis study illustrated that LNR was independent prognostic factor of DFS and OS in MTC. In addition, LNR influenced biochemical cure. Further investigations are needed to determine the optimal cut-off value for predicting prognosis. creator: Weijing Hao creator: Jingzhu Zhao creator: Fengli Guo creator: Pengfei Gu creator: Jinming Zhang creator: Dongmei Huang creator: Xianhui Ruan creator: Yu Zeng creator: Xiangqian Zheng creator: Ming Gao uri: https://doi.org/10.7717/peerj.15025 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Hao et al. title: Newly compiled Tai Chi (Bafa Wubu) promotes lower extremity exercise: a preliminary cross sectional study link: https://peerj.com/articles/15036 last-modified: 2023-03-13 description: BackgroundTai Chi (Bafa Wubu) is a new type of simplified Tai Chi widely practiced by Tai Chi enthusiasts that has developed and perfected simplified Tai Chi movement and enriched Tai Chi practice methods. When practicing, Tai Chi athletes and enthusiasts can choose the Bafa Wubu movements to practice according to their physical conditions. The purpose of this article is to discuss the mechanism by which Bafa Wubu promotes lower extremity exercise from the perspective of exercise biomechanics.ObjectivesThis article aims to explore the scientific training methods and technical characteristics of Bafa Wubu, and its contribution to comprehensive exercise of the lower extremities, by analyzing the biomechanical characteristics of the lower extremities of participants who practice Bafa Wubu at different levels and by comparing their ground reaction force, lower limb joints, and muscles during Bafa Wubu.MethodsA total of 16 male participants were recruited and divided into an amateur group (N = 8) and a professional group (N = 8). The data were collected by a BTS 3D infrared-based motion capture system, and Kistler 3D force plate. The lower extremity joint forces and muscle strength were calculated by anybody simulation software with inverse dynamics.ResultsDuring elbowing and leaning sideways with steps sideways (ELS), the ground reaction force of the professional group was significantly higher than that of the amateur group in the sagittal, vertical, and frontal axes (P < 0.01). While stepping forward, backward, and sideways, the professional group’s joints loading at the hip, knee, and ankle was always higher in the vertical direction (P < 0.01). Furthermore, during warding off with steps forward (WOF), laying with steps forward (LF), and rolling back with steps backward (RBB), hip joint loading increased in the med–lat direction. During actions with steps backward and sideways, the professional group’s ankle flexion/extension torque and hip abduction/rotation torque were significantly larger than those of the amateur group (P < 0.01). Different actions in Bafa Wubu activate muscles to different degrees, whereas the iliacus is mainly responsible for stabilizing postures when practitioners perform standing knee lifting motions.ConclusionsProfessional groups who have been practicing Tai Chi (Bafa Wubu) for a long time have higher ground reaction force, and the force on the three joints of the lower extremities is different for various movements, which has positive significance for exercising the joints of the lower extremities. In addition, various motions activate muscles of different types at different levels. For amateurs to practice different movements to stimulate the muscles, targeted areas of practice promote the lower extremity muscles’ synergistic force. In summary, the muscles and joints of the lower extremity can obtain comprehensive and balanced exercise through Bafa Wubu. creator: Haojie Li creator: Fang Peng creator: Shaojun Lyu creator: Zhongqiu Ji creator: Xiongfeng Li creator: Mingyu Liu uri: https://doi.org/10.7717/peerj.15036 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Li et al. title: Acute ventilatory responses to swimming at increasing intensities link: https://peerj.com/articles/15042 last-modified: 2023-03-13 description: BackgroundPhysical exercise is a source of stress to the human body, triggering different ventilatory responses through different regulatory mechanisms and the aquatic environment imposes several restrictions to the swimmer, particularly regarding the restricted ventilation. Thus, we aimed to assess the acute ventilatory responses and to characterize the adopted breathing patterns when swimming front crawl at increasing intensity domains.MethodsEighteen well-trained swimmers performed 7 × 200 m front crawl (0.05 m∙s−1 velocity increments) and a maximal 100 m (30 s rest intervals). Pulmonary gas exchange and ventilation were continuously measured (breath-by-breath) and capillary blood samples for lactate concentration ([La−]) analysis were collected at rest, during intervals and at the end of the protocol, allowing the identification of the low, moderate, heavy, severe and extreme intensity domains.ResultsWith the swimming velocity rise, respiratory frequency (fR), [La−] and stroke rate (SR) increased ([29.1–49.7] breaths∙min−1, [2.7–11.4] mmol∙L−1, [26.23–40.85] cycles; respectively) and stroke length (SL) decreased ([2.43–2.04] m∙min−1; respectively). Oxygen uptake (VO2), minute ventilation (VE), carbon dioxide production (VCO2) and heart rate (HR) increased until severe ([37.5–53.5] mL∙kg−1∙min−1, [55.8–96.3] L∙min−1, [32.2–51.5] mL∙kg−1∙min−1 and [152–182] bpm; respectively) and stabilized from severe to extreme (53.1 ± 8.4, mL∙kg−1∙min-1, 99.5 ± 19.1 L∙min−1, 49.7 ± 8.3 mL∙kg−1∙min−1 and 186 ± 11 bpm; respectively) while tidal volume (VT) was similar from low to severe ([2.02–2.18] L) and decreased at extreme intensities (2.08 ± 0.56 L). Lastly, the fR/SR ratio increased from low to heavy and decreased from severe to the extreme intensity domains (1.12 ± 0.24, 1.19 ± 0.25, 1.26 ± 0.26, 1.32 ± 0.26 and 1.23 ± 0.26).ConclusionsOur findings confirm a different ventilatory response pattern at extreme intensities when compared to the usually evaluated exertions. This novel insight helps to understand and characterize the maximal efforts in swimming and reinforces the importance to include extreme efforts in future swimming evaluations. creator: Ana Sofia Monteiro creator: José Francisco Magalhães creator: Beat Knechtle creator: Cosme F. Buzzachera creator: J. Paulo Vilas-Boas creator: Ricardo J. Fernandes uri: https://doi.org/10.7717/peerj.15042 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Monteiro et al. title: Natural durability of timber species exposed to xylophagous fungi in southern Durango, Mexico link: https://peerj.com/articles/14541 last-modified: 2023-03-10 description: IntroductionWood is a natural resource used for construction and the manufacture of many products. This material is exposed to damage due to biotic and abiotic factors. An important biotic factor is wood-degrading fungi that generate large economic losses. The objectives of this study were to determine the effect of xylophagous fungi (Coniophora puteana and Trametes versicolor) on the natural durability of six timber species in southern Durango, Mexico, and to establish differences between fungal effects on each tree species.Materials and MethodsSamples of Pinus durangensis, P. cooperi, P. strobiformis, Juniperus deppeana, Quercus sideroxyla, and Alnus acuminata were exposed to fungi for 4 months under laboratory conditions according to European Standard EN350-1. Samples of Fagus sylvatica were used as control. Durability was determined as the percentage of wood mass loss for each species. Welch ANOVA tests were performed to establish differences among tree species. Welch t-tests were used to prove loss mass differences between fungi for each tree species.ResultsThe most resistant species to C. puteana were P. durangensis, J. deppeana, P. cooperi and P. strobiformis, showing mean mass losses lower than 8.08%. The most resistant species to T. versicolor were J. deppeana, P. strobiformis and P. durangensis (mean mass losses lower than 7.39%). Pinus strobiformis and Q. sideroxyla were more susceptible to C. puteana effect; in contrast, P. durangensis and P. cooperi showed more damage due to T. versicolor degradation.ConclusionsWoods of P. durangensis, P. cooperi, P. strobiformis and Juniperus deppeana are well adapted to infection by these xylophagous fungi and are therefore highly recommended for commercial use in southern Durango, Mexico. creator: Yolanda Ontiveros-Moreno creator: Serafín Colín-Urieta creator: José Javier Corral-Rivas creator: José Ciro Hernández-Díaz creator: José Ángel Prieto-Ruíz creator: Artemio Carrillo-Parra uri: https://doi.org/10.7717/peerj.14541 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Ontiveros-Moreno et al. title: Drought stress resistance indicators of chickpea varieties grown under deficit irrigation conditions link: https://peerj.com/articles/14818 last-modified: 2023-03-10 description: The aim of this study was to determine the drought stress resistance of three chickpea cultivars (Inci, Hasanbey and Seçkin) grown under water deficit conditions and to discuss the use of yield, crop water stress index and chlorophyll index values as drought stress tolerance indicators in breeding studies. Three drought stress levels, (full irrigation = no stress - I100, deficit irrigation = moderate stress - I50, and no irrigation = severe stress - I0) were used as irrigation treatments. The highest seed yield (1,984 kg ha−1) in severe stress conditions was recorded for the Inci cultivar with a low crop water stress index (CWSI) (0.50) and high chlorophyll index (33.60 SPAD). The lowest seed yield (1,783.66 kg ha−1) in I0treatment was noted for the Seçkin cultivar which had a high CWSI (0.58) and low chlorophyll index (32.88 SPAD). The highest seed yield (2,566.33 kg ha−1) in full irrigation was recorded for the Inci cultivar which had a low CWSI (0.19) and high chlorophyll index (44.39 SPAD), while the lowest seed yield (2,328.00 kg ha−1) in I100 treatment was recorded for the Seçkin cultivar which had a high CWSI (0.26) and low chlorophyll index (42.12 SPAD). The seed yield of the Hasanbey cultivar in both severe stress (1,893 kg ha−1) and full irrigation (2,424.00 kg ha−1) conditions was between Inci and Seçkin varieties. The chlorophyll index and yield had a significant positive (r = 0.877) correlation, while a significant negative (r = −0.90) relationship was determined between CWSI and yield. Seed yield of the Inci cultivar in I0and I100treatments and water use efficiency revealed that the Inci cultivar is resistant to drought stress. Therefore, the Inci cultivar can be used in drought stress tolerance studies. In addition, the CWSI and chlorophyll index values can be employed as resistance indicators in chickpea breeding studies to determine the drought resistant chickpea cultivars. creator: Ali Beyhan Ucak creator: Hüseyin Arslan uri: https://doi.org/10.7717/peerj.14818 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Ucak and Arslan title: LTA4H extensively associates with mRNAs and lncRNAs indicative of its novel regulatory targets link: https://peerj.com/articles/14875 last-modified: 2023-03-10 description: The RNA-binding metabolic enzyme LTA4H is a novel target for cancer chemoprevention and chemotherapy. Recent research shows that the increased expression of LTA4H in laryngeal squamous cell carcinoma (LSCC) promotes tumor proliferation, migration, and metastasis. However, its mechanism remains unclear. To investigate the potential role of LTA4H in LSCC, we employed the improved RNA immunoprecipitation and sequencing (iRIP-Seq) experiment to get the expression profile of LTA4H binding RNA in HeLa model cells, a cancer model cell that is frequently used in molecular mechanism research. We found that LTA4H extensively binds with mRNAs/pre-mRNAs and lncRNAs. In the LTA4H binding peak, the frequency of the AAGG motif reported to interact with TRA2β4 was high in both replicates. More notably, LTA4H-binding genes were significantly enriched in the mitotic cell cycle, DNA repair, RNA splicing-related pathways, and RNA metabolism pathways, which means that LTA4H has tumor-related alternative splicing regulatory functions. QRT-PCR validation confirmed that LTA4H specifically binds to mRNAs of carcinogenesis-associated genes, including LTBP3, ROR2, EGFR, HSP90B1, and lncRNAs represented by NEAT1. These results suggest that LTA4H may combine with genes associated with LSCC as an RNA-binding protein to perform a cancer regulatory function. Our study further sheds light on the molecular mechanism of LTA4H as a clinical therapy target for LSCC. creator: Tianjiao Ren creator: Song Wang creator: Bo Zhang creator: Wei Zhou creator: Cansi Wang creator: Xiaorui Zhao creator: Juan Feng uri: https://doi.org/10.7717/peerj.14875 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Ren et al. title: Modelling quarantine effects on SARS-CoV-2 epidemiological dynamics in Chilean communes and their relationship with the Social Priority Index link: https://peerj.com/articles/14892 last-modified: 2023-03-10 description: BackgroundAn epidemiological model (susceptible, un-quarantined infected, quarantined infected, confirmed infected (SUQC)) was previously developed and applied to incorporate quarantine measures and calculate COVID-19 contagion dynamics and pandemic control in some Chinese regions. Here, we generalized this model to incorporate the disease recovery rate and applied our model to records of the total number of confirmed cases of people infected with the SARS-CoV-2 virus in some Chilean communes.MethodsIn each commune, two consecutive stages were considered: a stage without quarantine and an immediately subsequent quarantine stage imposed by the Ministry of Health. To adjust the model, typical epidemiological parameters were determined, such as the confirmation rate and the quarantine rate. The latter allowed us to calculate the reproduction number.ResultsThe mathematical model adequately reproduced the data, indicating a higher quarantine rate when quarantine was imposed by the health authority, with a corresponding decrease in the reproduction number of the virus down to values that prevent or decrease its exponential spread. In general, during this second stage, the communes with the lowest social priority indices had the highest quarantine rates, and therefore, the lowest effective viral reproduction numbers. This study provides useful evidence to address the health inequity of pandemics. The mathematical model applied here can be used in other regions or easily modified for other cases of infectious disease control by quarantine. creator: Dino G. Salinas creator: M. Leonor Bustamante creator: Mauricio O. Gallardo uri: https://doi.org/10.7717/peerj.14892 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Salinas et al. title: HNF4G increases cisplatin resistance in lung adenocarcinoma via the MAPK6/Akt pathway link: https://peerj.com/articles/14996 last-modified: 2023-03-10 description: BackgroundLung adenocarcinoma is one of the most common tumors, and cisplatin is frequently used in treating lung adenocarcinoma patients. This study aimed to look into the roles and mechanisms of HNF4G in cisplatin resistance of lung adenocarcinoma.Materials & MethodsCisplatin resistance and gene expression data of 542 cell lines from the CTRP and CCLE databases were analyzed. HNF4G expression was detected in the lung adenocarcinoma cell lines after treatment with various concentrations of cisplatin. Cisplatin sensitivity curves were detected in cells that overexpressed or knocked down HNF4G. The ChIP-Seq data were then analyzed to identify the targets of HNF4G involved in cisplatin resistance. Expression and phosphorylation of the MAPK6/Akt pathway were detected after HNF4G was overexpressed or knocked down. Finally, ChIP-qPCR and dual-luciferase assays were used to investigate the regulation of HNF4G on MAPK6.ResultsIn cell lines, high expression of HNF4G was significantly positively correlated with cisplatin resistance, and lung adenocarcinoma patients who had high HNF4G expression had a poor prognosis. Cisplatin treatment increased HNF4G expression, and overexpression of HNF4G significantly increased the resistance to cisplatin in A549 and HCC827 cells, whereas knockdown of HNF4G had the opposite effect. HNF4G overexpression increased MAPK6 expression and activated the MAPK6/Akt pathway, while an Akt inhibitor reduced the effects of HNF4G on cisplatin resistance. HNF4G bound to the MAPK6 promoter region, promoting MAPK6 expression, according to ChIP-qPCR and luciferase assays.ConclusionBy binding to the MAPK6 promoter region, HNF4G promotes MAPK6 expression and subsequent Akt phosphorylation, resulting in resistance to cisplatin in lung adenocarcinoma. creator: Jiaqi Liang creator: Guangyin Zhao creator: Yunyi Bian creator: Guoshu Bi creator: Qihai Sui creator: Huan Zhang creator: Haochun Shi creator: Guangyao Shan creator: Yiwei Huang creator: Zhencong Chen creator: Lin Wang creator: Cheng Zhan uri: https://doi.org/10.7717/peerj.14996 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Liang et al. title: Paranoid thinking and perceived competitive intention link: https://peerj.com/articles/15003 last-modified: 2023-03-10 description: Paranoid thinking, that others are hostile, can be seen even in the general population. Paranoia is considered the expectation that others are competitors who aim to maximize the differences in payoffs rather than maximize their own payoffs. This study examined whether paranoia reflects the irrational belief that others have a competitive intention and is associated with avoiding perceived competition. We recruited 884 US residents via the Internet and conducted a modified Dictator Game, in which monetary allocation was carried out between the Dictator and the Recipient. The Dictator chooses either fair or competitive allocation while selecting the competitive allocation is irrelevant to increasing the Dictator’s payoffs. The Recipient decides whether to accept the Dictator’s decision or receive sure but low rewards. We found that Recipients with high-level paranoid thinking expected their opponent to select competitive allocation more than those with low levels, even when selecting it was costly for Dictators. Paranoid thinking was not associated with selecting sure rewards or competitive allocations. The results suggest that paranoia reflects the belief that others have a competitive intention but is not related to avoidance behavior against perceived threats and unilateral attacks. creator: Yutaka Horita uri: https://doi.org/10.7717/peerj.15003 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Horita title: Survival of Pseudomonas syringae pv. actinidiae in detached kiwifruit leaves at different environmental conditions link: https://peerj.com/articles/15031 last-modified: 2023-03-10 description: Pseudomonas syringae pv. actinidiae (Psa) is the causal agent of kiwifruit canker, a serious threat to commercial kiwifruit production worldwide. Studies of the movement path and the survival time of Psa in the host are crucial for integrated management programs. Hence, we used Psa with GFPuv gene (Psa-GFPuv) strain to investigate the movement path of Psa in leaves and branches, and the survival time of Psa in leaves under different environmental conditions. We found that the pathogen Psa spread longitudinally in the branches and leaves rather than transverse path. Additionally, the survival time of bacteria in fallen leaves under different environmental conditions were simulated by the way of Psa infecting the detached kiwifruit leaves. Psa survives the longest, up to 43 days in detached kiwifruit leaves with high humidity (above 80%) at 5 °C, and up to 32 days with low humidity (20%). At 15 °C, the Psa can survive in detached kiwifruit leaves for 20–30 days with increasing humidity. At 25 °C, it can only survive for 3 days with low humidity (20%) and 15 days with high humidity (above 80%). Furthermore, the population growth experiments showed that bacterial growth of Psa was more favorable in detached kiwifruit leaves with above 80% humidity at 5 °C. These results suggest that the survival condition of Psa in detached kiwifruit leaves is significantly affected by environmental conditions, and provide the basis for the control timing and technology of kiwifruit canker. creator: Siyi Deng creator: Wei Chang creator: Yawei Que creator: Jun Liu creator: Hua Wang uri: https://doi.org/10.7717/peerj.15031 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Deng et al. title: Anti-leukemic effect of menthol, a peppermint compound, on induction of apoptosis and autophagy link: https://peerj.com/articles/15049 last-modified: 2023-03-10 description: BackgroundMenthol, a natural compound in peppermint leaves, has several biological activities, including antioxidant, anti-inflammatory, antiviral, antibacterial and anticancer properties. This study revealed the anti-leukemic effects and its underlying mechanisms of the menthol related apoptosis signaling pathway and autophagy in both NB4 and Molt-4 leukemic cell lines.MethodsBoth leukemic cells were treated with menthol in various concentration. Cell viability was assessed using MTT assay, whereas apoptosis and autophagy were analyzed by flow cytometry using Annexin V-FITC/PI and anti-LC3/FITC antibodies staining, respectively. Apoptotic and autophagic related gene and protein expression were detected using RT-qPCR and western blot analysis, respectively. Moreover, STITCH database was used to predicts the interaction between menthol and proposed proteins.ResultsMenthol significantly decreased cell viability in NB4 and Molt-4 cell lines in dose dependent manner. In combination of menthol and daunorubicin, synergistic cytotoxic effects were observed in leukemic cells. However, there was a minimal effect found on normal, peripheral blood mononuclear cells (PBMCs). Moreover, menthol significantly induced apoptosis induction via upregulation of caspase-3, BAX, p53 and downregulation of MDM2 mRNA expression. Autophagy was also induced by menthol through upregulating ATG3 and downregulating mTOR mRNA expression. For protein expression, menthol significantly increased caspase-3 whereas decreased mTOR in both leukemic cells. Conclusions. These results suggest that menthol exhibits cytotoxic activities by inhibition of cell proliferation, induction of apoptosis and autophagy through activating the caspase cascade, altering BAX and p53/MDM2, and regulating autophagy via the ATG3/mTOR signaling pathway. creator: Mashima Naksawat creator: Chosita Norkaew creator: Kantorn Charoensedtasin creator: Sittiruk Roytrakul creator: Dalina Tanyong uri: https://doi.org/10.7717/peerj.15049 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Naksawat et al. title: Screening plant growth-promoting bacteria from the rhizosphere of invasive weed Ageratina adenophora for crop growth link: https://peerj.com/articles/15064 last-modified: 2023-03-10 description: Plant-growth promoting rhizobacteria (PGPR) play a vital role in soil fertility and crop production. The rhizosphere of many crop plants has been well documented by screening PGPR for their plant-growth promoting (PGP) mechanisms. However, the rhizosphere of grass species that may act as potential habitats for novel PGPR remains relatively unexplored. Ageratina adenophora is a noxious weed that has invaded more than 40 tropical and subtropical countries in Asia, Oceania, Africa, and Europe. Its presence has led to changes in plant species composition, reducing their biodiversity and destroying ecosystem function. In this study, we screened 1,200 bacterial strains isolated from the rhizosphere soil of A. adenophora in three floristic regions in Yunnan Province, China. Samples were screened for their in vitro ability for N-fixation, production of the plant growth regulator indole-3-acetic acid (IAA), and the synthesis of 1-amino-cyclopropane-1-carboxylate (ACC) deaminase, which controls the levels of ethylene in developing plant roots. We found that 144 strains showed at least one of these PGP attributes. 16S rRNA gene sequencing showed that most (62.5%) of the samples were bacteria closely related to members of the genera Pseudomonas (27 strains), Providencia (20 strains), Chryseobacterium (14 strains), Ensifer (12 strains), Enterobacter (nine strains), and Hafnia (eight strains). Their abundance and biodiversity in the soil of individual floristic regions correlate positively with the invasion history of A. adenophora. From these PGP bacterial strains, KM_A34 (Pantoea agglomerans), KM_C04 (Enterobacter asburiae), and KM_A57 (Pseudomonas putida), which had the greatest in vitro ability of N-fixation, and IAA and ACC deaminase production, respectively, were selected. The strains were evaluated for their effect on the seed germination and growth of soybean, faba bean, pea, wheat, and Chinese cabbage other than A. adenophora. Chamber experiments showed these strains significantly (P < 0.05) increased (14.2–43.4% over the controls) germination rates of the soybean, faba bean, pea, and/or Chinese cabbage seeds. They also reduced relative seed germination times (20.8–48.8% over the controls) of soy bean, faba bean and/or wheat seeds. Greenhouse pot experiments showed that they significantly (P < 0.05) promoted the aboveground and belowground height of plant foliage (12.1–23.1% and 11.5–31.4% over the controls, respectively) and/or the dry weights (16.1–33.5% and 10.6–23.4% over the controls, respectively) of the soy bean, faba bean, pea, wheat and/or Chinese cabbage. These data indicate that the rhizosphere microbiota of A. adenophora contain a PGPR pool that may be used as bioinoculants to improve the growth and productivity of these crops. creator: Yun Xia creator: Hongbo Zhang creator: Yu Zhang creator: Yuyu Zhang creator: Jiani Liu creator: Robert Seviour creator: Yunhong Kong uri: https://doi.org/10.7717/peerj.15064 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Xia et al. title: Comprehensive analysis of COMMD10 as a novel prognostic biomarker for gastric cancer link: https://peerj.com/articles/14645 last-modified: 2023-03-09 description: BackgroundCOMMD10 has an important role in the development of certain tumors, but its relevance to gastric cancer (GC) is unclear. The purpose of this study is to investigate the difference of COMMD10 expression in gastric adenocarcinoma (STAD) and analyze the correlation between COMMD10 expression and prognosis of STAD patients.MethodsThe expression levels of COMMD10 between STAD and normal tissues were explored using the The Cancer Genome Atlas (TCGA) database. In addition, the expression of COMMD10 in GC was further validated by immunohistochemistry (IHC) staining, qRT-PCR and Western blot. Dot blot experiments were used for exploring m6A expression levels in tissues with high and low COMMD10 expression. Kaplan–Meier analysis and COX regression analysis were used to explore the relationship between COMMD10 and STAD prognosis. A nomogram was constructed to predict the survival probability of STAD patients. GO and KEGG functional enrichment of COMMD10-related genes were performed. The Corrlot software package was used to analyze the correlation between COMMD10 expression levels and m6A modifications in STAD. An analysis of immune infiltration based on the CIBERSOFT and the single-sample GSEA (ssGSEA) method was performed.ResultsCOMMD10 expression was significantly associated with multiple cancers, including STAD in TCGA. COMMD10 expression was elevated in STAD cancer tissues compared to paracancerous tissues. COMMD10 upregulation was associated with poorer overall survival (OS), clinical stage, N stage, and primary treatment outcome in STAD. Functional enrichment of COMMD10-related genes was mainly involved in biological processes such as RNA localization, RNA splicing, RNA transport, mRNA surveillance pathways, and spliceosomes. The dot blot experiment showed that m6A levels were higher in cancer tissues with high COMMD10 expression compared with paracancerous tissues. COMMD10 was significantly correlated with most m6A-related genes. COMMD10 was involved in STAD immune cells infiltration, correlated with macrophage cells expression.ConclusionHigh COMMD10 expression was significantly associated with poor prognosis in STAD patients, and its functional realization was related to m6A modification. COMMD10 involved in STAD immune infiltration. creator: Wenfang Zhao creator: Jiahui Lin creator: Sha Cheng creator: Huan Li creator: Yufeng Shu creator: Canxia Xu uri: https://doi.org/10.7717/peerj.14645 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Zhao et al. title: Analysis of Corynebacterium silvaticum genomes from Portugal reveals a single cluster and a clade suggested to produce diphtheria toxin link: https://peerj.com/articles/14895 last-modified: 2023-03-09 description: BackgroundCorynebacterium silvaticum is a pathogenic, gram-positive bacterial species that causes caseous lymphadenitis in wild boars, domestic pigs and roe deer in Western Europe. It can affect animal production and cause zoonosis. Genome analysis has suggested that one strain from Portugal and one from Austria could probably produce the diphtheria toxin (DT), which inhibits protein synthesis and can cause death.MethodsTo further investigate the species genetic diversity and probable production of DT by Portuguese strains, eight isolates from this country were sequenced and compared to 38 public ones.ResultsStrains from Portugal are monophyletic, nearly identical, form a unique cluster and have 27 out of 36 known Corynebacterium virulence or niche factors. All of them lack a frameshift in the tox gene and were suggested to produce DT. A phylogenetic analysis shows that the species has diverged into two clades. Clade 1 is composed of strains that were suggested to have the ability to produce DT, represented by the monophyletic strains from Portugal and strain 05-13 from Austria. Clade 2 is composed of strains unable to produce DT due to a frameshifted tox gene. The second clade is represented by strains from Austria, Germany and Switzerland. Ten genome clusters were detected, in which strains from Germany are the most diverse. Strains from Portugal belong to an exclusive cluster. The pangenome has 2,961 proteins and is nearly closed (α = 0.968). Exclusive genes shared by clusters 1 and 2, and Portuguese strains are probably not related to disease manifestation as they share the same host but could play a role in their extra-host environmental adaptation. These results show the potential of the species to cause zoonosis, possibly diphtheria. The identified clusters, exclusively shaded genes, and exclusive STs identified in Portugal could be applied in the identification and epidemiology of the species. creator: Marcus Vinicius Canario Viana creator: José Henrique Galdino creator: Rodrigo Profeta creator: Manuela Oliveira creator: Luís Tavares creator: Siomar de Castro Soares creator: Paulo Carneiro creator: Alice Rebecca Wattam creator: Vasco Azevedo uri: https://doi.org/10.7717/peerj.14895 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Viana et al. title: Influences of environment, human activity, and climate on the invasion of Ageratina adenophora (Spreng.) in Southwest China link: https://peerj.com/articles/14902 last-modified: 2023-03-09 description: With economic and social globalization, invasive alien species have significantly threatened local ecological security. Identifying the invasive mechanisms of invasive alien species can aid in preventing species invasions and protecting local ecological and economic security. As a globally invasive plant, Ageratina adenophora (Asteraceae) has spread to many parts of the world and had a seriously impacted the ecology and economy of its invaded areas. Using observational data and Landsat OLI images in an arid valley region in southwest China, this study examined how climate, human activity and environmental factors influence the invasion of A. adenophora and its underlying mechanism. Our results showed that the invasion abundance of A. adenophora was significantly affected by environmental factors (the relative importance was 87.2%), but was less influenced by human activity and climate factors (the relative importance was 2% and 10.8%, respectively). The A. adenophora abundance significantly decreased with aspect, community canopy density, shrub layer coverage, herb layer coverage, Simpson diversity index of shrub and herb layers, the shortest distance to residential areas and temperature seasonality, whereas it increased with soil moisture, temperature annual range, precipitation of wettest month and precipitation of driest month. We conclude that biotic competition is the most influential factor in the invasion of this plant in the arid valley regions. Our results are of great significance for invasion prevention and forest conservation and management in southwest China. Our work emphasized that optimizing the community structure, such as by increasing canopy and shrub coverage and species biodiversity, may help control and mitigate the A. adenophora invasion in southwest China. creator: Xiaojuan Zhang creator: Guoyan Wang creator: Peihao Peng creator: Yongxiu Zhou creator: Zhuo Chen creator: Yu Feng creator: Yanru Wang creator: Songlin Shi creator: Jingji Li uri: https://doi.org/10.7717/peerj.14902 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Zhang et al. title: Reporting and interpreting non-significant results in animal cognition research link: https://peerj.com/articles/14963 last-modified: 2023-03-09 description: How statistically non-significant results are reported and interpreted following null hypothesis significance testing is often criticized. This issue is important for animal cognition research because studies in the field are often underpowered to detect theoretically meaningful effect sizes, i.e., often produce non-significant p-values even when the null hypothesis is incorrect. Thus, we manually extracted and classified how researchers report and interpret non-significant p-values and examined the p-value distribution of these non-significant results across published articles in animal cognition and related fields. We found a large amount of heterogeneity in how researchers report statistically non-significant p-values in the result sections of articles, and how they interpret them in the titles and abstracts. Reporting of the non-significant results as “No Effect” was common in the titles (84%), abstracts (64%), and results sections (41%) of papers, whereas reporting of the results as “Non-Significant” was less common in the titles (0%) and abstracts (26%), but was present in the results (52%). Discussions of effect sizes were rare (<5% of articles). A p-value distribution analysis was consistent with research being performed with low power of statistical tests to detect effect sizes of interest. These findings suggest that researchers in animal cognition should pay close attention to the evidence used to support claims of absence of effects in the literature, and—in their own work—report statistically non-significant results clearly and formally correct, as well as use more formal methods of assessing evidence against theoretical predictions. creator: Benjamin G. Farrar creator: Alizée Vernouillet creator: Elias Garcia-Pelegrin creator: Edward W. Legg creator: Katharina F. Brecht creator: Poppy J. Lambert creator: Mahmoud Elsherif creator: Shannon Francis creator: Laurie O’Neill creator: Nicola S. Clayton creator: Ljerka Ostojić uri: https://doi.org/10.7717/peerj.14963 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Farrar et al. title: Combining ability, heterosis and performance of grain yield and content of Fe, Zn and protein in bread wheat under normal and late sowing conditions link: https://peerj.com/articles/14971 last-modified: 2023-03-09 description: Wheat (Triticum aestivum L.) is inherently low in protein content, Zn and Fe. Boost yield gains have unwittingly reduced grain Zn and Fe, which has had negative impacts on human health. The aim of this study was to understand the inheritance of grain yield per plant and grain Fe, Zn, and protein concentrations in bread wheat (Triticum aestivum L.) under normal and late sown conditions. Half diallel crosses were performed using 10 parents. The crosses and parents were evaluated in replicated trials for the two conditions, to assess the possibility of exploiting heterosis to improve micronutrient contents. The per se performance, heterosis, combining ability, and genetic components were estimated for different characters in both environments. The results revealed that hybrid GW 451 × GW 173 exhibited better parent heterosis (BPH) and standard heterotic effects (SH) in all environments. In both sowing conditions, the general combining ability (GCA) effects of poor × poor parents also showed high specific combining ability (SCA) effects of hybrids for both the micronutrients and protein contents. However, σ2A/σ2D greater than unity confirmed the preponderance of additive gene action for protein content, and GW 173 was identified as a good general combiner for these characteristics under both environments. SCA had positive significant (P < 0.001) correlations with BPH, SH1, SH2, and the phenotype for yield component traits and grain protein, Fe, and Zn concentrations in both conditions. A supplementary approach for biofortifying wheat grainis required to prevent malnutrition. creator: Gita R. Chaudhari creator: D. A. Patel creator: D. J. Parmar creator: K. C. Patel creator: Sushil Kumar uri: https://doi.org/10.7717/peerj.14971 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Chaudhari et al. title: Effects of stumping on fine root architecture, growth, and physiology of Hippophae rhamnoides link: https://peerj.com/articles/14978 last-modified: 2023-03-09 description: BackgroundFine roots are vital to a plant’s ability to absorb water and nutrients. Stumping is a practice that may encourage fine root growth and the rapid recovery of decaying Hippophae rhamnoides plants. However, the effect of stumping on the fine roots and physiological indices is still unknown. The differential indices between stumped forests and non-stumped forests must also be defined.MethodsWe recorded the changes in the fine roots of structure H. rhamnoides one year after stumping. Using single factor analysis of variance and general linear models we comprehensively analyzed the number of root tips and the plant’s growth and physiological indices in response to stumping. Partial least squares discriminant analysis (PLS-DA) was used to compare fine root growth and physiological indices with and without stumping in order to identify the differential indices.ResultsThe proportion of root tips in the vertical layers at 30–40 cm and 40–50 cm and in the horizontal layers at 60–80 cm and 80–100 cm, increased after stumping by 1.85%, 2.60%, 1.96% and 4.32%, respectively. In the 0–50 cm soil layer, the fine root dry weight rose by 27.6% compared with the control, which was not significant. However, other indices were significantly different from the control. The proportions in the growth indices in the 30–40 cm and 40–50 cm layers increased after stumping. Stumping had a significant, negative effect on proline and malondialdehyde content, which dropped by 40.95% and 55.32%, respectively, indicating that the harms caused by these two chemicals was alleviated. Stumping had a significant positive effect on root activity and soluble sugar contents, which increased by 68.58% and 36.87%, respectively, and improved the growing ability of fine roots. PLS-DA revealed that malondialdehyde, soluble sugars, root density, and the number of root tips ranked from having the least to greatest effect on the classification of stumping and no-stumping.ConclusionsThe process of stumping may promote fine roots growth in H. rhamnoides, and is favorable for their longitudinal development. The fine root growing indices of H. rhamnoides responded positively to this process. Stumping promotes root activity and the creation of soluble sugar to maintain the growth and development of fine roots. It also inhibits the negative effects of proline and malondialdehyde on fine roots. Our study showed that the differential physiological indices were more important for classification than the differential growing indices. creator: Haoyue Wang creator: Wei Qi creator: Yuefeng Guo creator: Yajie Xu uri: https://doi.org/10.7717/peerj.14978 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Wang et al. title: Ensemble modeling of black pomfret (Parastromateus niger) habitat in the Taiwan Strait based on oceanographic variables link: https://peerj.com/articles/14990 last-modified: 2023-03-09 description: The location, effort, number of captures, and time of fishing were all used in this study to assess the geographic distribution of Parastromateus niger in the Taiwan Strait. Other species distribution models performed worse than generalized linear models (GLMs) based on six oceanographic parameters. The sea surface temperature (SST) was between 26.5 °C and 29.5 °C, the sea surface chlorophyll (SSC) level was between 0.3–0.44 mg/m3, the sea surface salinity (SSS) was between 33.4 °C and 34.4 °C, the mixed layer depth was between 10 °C and 14 °C, the sea surface height was between 0.57 °C and 0.77 °C, and the eddy kinetic energy (EKE) was between 0.603 °C. According to the statistical findings, SST is merely a small effect compared to SSS, SSC level, and EKE in terms of impacting species distribution. By combining four effective single-algorithm models with no obvious bias, an ensemble habitat model was created. The ranges of 117°E–119°E and 22°N–24°N have the highest annual distributions of S.CPUE and nominal CPUE. creator: Sandipan Mondal creator: Ming An Lee creator: Yu-Kai Chen creator: Yi-Chen Wang uri: https://doi.org/10.7717/peerj.14990 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Mondal et al. title: Intra- and inter-session reliability and repeatability of an infrared thermography device designed for materials to measure skin temperature of the triceps surae muscle tissue of athletes link: https://peerj.com/articles/15011 last-modified: 2023-03-09 description: BackgroundInfrared thermography devices have been commonly applied to measure superficial temperature in structural composites and walls. These tools were cheaper than other thermographic devices used to measure superficial human muscle tissue temperature. In addition, infrared thermography has been previously used to assess skin temperature related to muscle tissue conditions in the triceps surae of athletes. Nevertheless, the reliability and repeatability of an infrared thermography device designed for materials, such as the Manual Infrared Camera PCE-TC 30, have yet to be determined to measure skin temperature of the triceps surae muscle tissue of athletes.ObjectiveThe purpose was to determine the procedure’s intra- and inter-session reliability and repeatability to determine skin temperature within the Manual Infrared Camera PCE-TC 30 thermography device in the triceps surae muscle tissue of athletes, which was initially designed to measure the superficial temperature of materials.MethodsA total of 34 triceps surae muscles were bilaterally assessed from 17 healthy athletes using the Manual Infrared Camera PCE-TC 30 thermography device to determine intra- (at the same day separated by 1 h) and inter-session (at alternate days separated by 48 h) reliability and repeatability of the skin temperature of the soleus, medial and lateral gastrocnemius muscles. The triceps surae complex weas measured by a region of interest of 1 cm2 through five infrared thermography images for each muscle. Statistical analyses comprised intraclass correlation coefficient (ICC), standard error of measurement (SEM), minimum detectable change (MCD), systematic error of measurement, correlation (r), and Bland-Altman plots completed with linear regression models (R2).ResultsIntra- and inter-session measurements of the proposed infrared thermography procedure showed excellent reliability (ICC(1,2) = 0.968–0.977), measurement errors (SEM = 0.186–0.232 °C; MDC = 0.515–0.643 °C), correlations (r = 0.885–0.953), and did not present significant systematic error of measurements (P > 0.05). Adequate agreement between each pair of measurement moments was presented by the Bland-Altman plots according to the limits of agreement and non-significant linear regression models (R2 = 0.000–0.019; P > 0.05).ConclusionsThe proposed procedure to determine skin temperature within the Manual Infrared Camera PCE-TC 30 thermography device presented excellent intra- and inter-session reliability and repeatability in athletes’ triceps surae muscle tissue. Future studies should consider the SEM and MDC of this procedure to measure the skin temperature of soleus, medial, and lateral gastrocnemius muscles to promote triceps surae muscle prevention and recovery in athletes. creator: Cesar Calvo-Lobo creator: Marta San-Antolín creator: Daniel García-García creator: Ricardo Becerro-de-Bengoa-Vallejo creator: Marta Elena Losa-Iglesias creator: Julia Cosín-Matamoros creator: Israel Casado-Hernández creator: Eva María Martínez-Jiménez creator: Victoria Mazoteras-Pardo creator: David Rodríguez-Sanz uri: https://doi.org/10.7717/peerj.15011 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Calvo-Lobo et al. title: In vitro and in silico evaluations of actinomycin X2and actinomycin D as potent anti-tuberculosis agents link: https://peerj.com/articles/14502 last-modified: 2023-03-08 description: BackgroundMultidrug-resistant tuberculosis (MDR-TB) is one of the world’s most devastating contagious diseases and is caused by the MDR-Mycobacterium tuberculosis (MDR-Mtb) bacteria. It is therefore essential to identify novel anti-TB drug candidates and target proteins to treat MDR-TB. Here, in vitro and in silico studies were used to investigate the anti-TB potential of two newly sourced actinomycins, actinomycin-X2 (act-X2) and actinomycin-D (act-D), from the Streptomyces smyrnaeus strain UKAQ_23 (isolated from the Jubail industrial city of Saudi Arabia).MethodsThe anti-TB activity of the isolated actinomycins was assessed in vitro using the Mtb H37Ra, Mycobacterium bovis (BCG), and Mtb H37Rv bacterial strains, using the Microplate Alamar Blue Assay (MABA) method. In silico molecular docking studies were conducted using sixteen anti-TB drug target proteins using the AutoDock Vina 1.1.2 tool. The molecular dynamics (MD) simulations for both actinomycins were then performed with the most suitable target proteins, using the GROningen MAchine For Chemical Simulations (GROMACS) simulation software (GROMACS 2020.4), with the Chemistry at HARvard Macromolecular Mechanics 36m (CHARMM36m) forcefield for proteins and the CHARMM General Force Field (CGenFF) for ligands.ResultsIn vitro results for the Mtb H37Ra, BCG, and Mtb H37Rv strains showed that act-X2 had minimum inhibitory concentration (MIC) values of 1.56 ± 0.0, 1.56 ± 0.0, and 2.64 ± 0.07 µg/mL and act-D had MIC values of 1.56 ± 0.0, 1.56 ± 0.0, and 1.80 ± 0.24 µg/mL respectively. The in silico molecular docking results showed that protein kinase PknB was the preferred target for both actinomycins, while KasA and pantothenate synthetase were the least preferred targets for act-X2and act-D respectively. The molecular dynamics (MD) results demonstrated that act-X2 and act-D remained stable inside the binding region of PknB throughout the simulation period. The MM/GBSA (Molecular Mechanics/Generalized Born Surface Area) binding energy calculations showed that act-X2 was more potent than act-D.ConclusionIn conclusion, our results suggest that both actinomycins X2 and D are highly potent anti-TB drug candidates. We show that act-X2is better able to antagonistically interact with the protein kinase PknB target than act-D, and thus has more potential as a new anti-TB drug candidate. creator: Kamal Ahmad Qureshi creator: Faizul Azam creator: Muhammad Qaiser Fatmi creator: Mahrukh Imtiaz creator: Dinesh Kumar Prajapati creator: Pankaj Kumar Rai creator: Mariusz Jaremko creator: Abdul-Hamid Emwas creator: Gamal Osman Elhassan uri: https://doi.org/10.7717/peerj.14502 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Qureshi et al. title: The earliest dipodomyine heteromyid in North America and the phylogenetic relationships of geomorph rodents link: https://peerj.com/articles/14693 last-modified: 2023-03-08 description: Dipodomyine heteromyids (kangaroo rats and mice) are a diverse group of arid-adapted ricochetal rodents of North America. Here, a new genus and species of a large dipodomyine is reported from early Miocene-aged deposits of the John Day Formation in Oregon that represents the earliest record of the subfamily. The taxon is known from a single specimen consisting of a nearly complete skull, dentary, partial pes, and caudal vertebra. The specimen is characterized by a mosaic of ancestral and highly derived cranial features of heteromyids. Specifically, the dental morphology and some cranial characteristics are similar to early heteromyids, but other aspects of morphology, including the exceptionally inflated auditory bullae, are more similar to known dipodomyines. This specimen was included in a phylogenetic analysis comprising 96 characters and the broadest sampling of living and extinct geomorph rodents of any morphological phylogenetic analysis to date. Results support the monophyly of crown-group Heteromyidae exclusive of Geomyidae and place the new taxon within Dipodomyinae. The new heteromyid is the largest known member of the family. Analyses suggest that large body size evolved several times within Heteromyidae. Overall, the morphology of the new heteromyid supports a mosaic evolution of the open-habitat adaptations that characterize kangaroo rats and mice, with the inflation of the auditory bulla appearing early in the group, and bipedality/ricochetal locomotion appearing later. We hypothesize that cooling and drying conditions in the late Oligocene and early Miocene favored adaptations for life in more open habitats, resulting in increased locomotor specialization in this lineage over time from a terrestrial ancestor. creator: Joshua X. Samuels creator: Jonathan J.-M. Calede creator: Robert M. Hunt, Jr. uri: https://doi.org/10.7717/peerj.14693 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Samuels et al. title: Long COVID active case detection initiative among COVID-19 patients in Port Dickson, Malaysia: a retrospective study on the positive outcomes, the proportion of patients with long COVID and its associated factors link: https://peerj.com/articles/14742 last-modified: 2023-03-08 description: BackgroundLong COVID is new or ongoing symptoms at four weeks or more after the start of acute COVID-19. However, the prevalence and factors associated with long COVID are largely unknown in Malaysia. We aim to determine the proportion and factors associated with long COVID among COVID-19 patients in Port Dickson, Malaysia. The positive outcomes of our long COVID active detection initiative were also described.MethodsThis was a retrospective analysis of long COVID data collected by the Port Dickson District Health Office between 1 September 2021 to 31 October 2021. Monitoring long COVID symptoms was our quality improvement initiative to safeguard residents’ health in the district. The study population was patients previously diagnosed with COVID-19 who resided in Port Dickson. The inclusion criteria were adults aged 18 years and above and were in the fifth week (day 29 to 35) post-COVID-19 diagnosis during the data collection period. We called all consecutive eligible patients to inquire regarding long COVID symptoms. Long COVID was defined as new or ongoing symptoms lasting more than 28 days from the date of positive SARS-CoV-2 by polymerase chain reaction test. Binary multivariate logistic regression was conducted to determine factors associated with long COVID.ResultsAmong 452 patients, they were predominantly male (54.2%), Malays (68.8%) and aged 18–29 years (58.6%). A total of 27.4% (95% CI [23.4–31.8]) of patients experienced long COVID symptoms and were referred to government clinics. The most frequent long COVID symptoms experienced were fatigue (54.0%), cough (20.2%), muscle pain (18.5%), headache (17.7%) and sleep disturbance (16.1%). Females, patients with underlying cardiovascular disease, asthma and chronic obstructive airway disease, those who received symptomatic care, and patients with myalgia and headaches at COVID-19 diagnosis were more likely to have long COVID. Three patients with suspected severe mental health problems were referred to the district psychologist, and ten patients with no/incomplete vaccination were referred for vaccination.ConclusionLong COVID is highly prevalent among COVID-19 patients in Port Dickson, Malaysia. Long-term surveillance and management of long COVID, especially among the high-risk groups, are needed as we transition to living with COVID-19. creator: Kim Sui Wan creator: Esther Rishma Sundram creator: Ammar Amsyar Abdul Haddi creator: Abdul Rahman Dashuki creator: Azainorsuzila Ahad creator: Rowena John creator: MuhammadKhairul Ridhuan Abdul Wahid creator: Ungku Izmin Farah Ungku Halmie creator: Farah Edura Ibrahim creator: Nachia Banu Abdul Rahim uri: https://doi.org/10.7717/peerj.14742 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Wan et al. title: Annotated checklist of the beetles (Coleoptera) of the California Channel Islands link: https://peerj.com/articles/14793 last-modified: 2023-03-08 description: The beetle fauna of the California Channel Islands is here enumerated for the first time in over 120 years. We provide an annotated checklist documenting species-by-island diversity from an exhaustive literature review and analysis of a compiled dataset of 26,609 digitized specimen records to which were added over 3,000 individual specimen determinations. We report 825 unique species from 514 genera and 71 families (including 17 new family records) comprising 1,829 species-by-island records. Species totals for each island are as follows: Anacapa (74); San Clemente (197); San Miguel (138); San Nicolas (146); Santa Barbara (64); Santa Catalina (370); Santa Cruz (503); and Santa Rosa (337). This represents the largest list of species published to date for any taxonomic group of animals on the Channel Islands; despite this, we consider the checklist to be preliminary. We present evidence that both inventory and taxonomic efforts on Channel Islands beetles are far from complete. Rarefaction estimates indicate there are at least several hundred more species of beetles yet to be recorded from the islands. Despite the incomplete nature of existing records, we found that species diversity is highly correlated with island area. We report 56 species which are putatively geographically restricted (endemic) to the Channel Islands, with two additional species of questionable endemic status. We also report 52 species from the islands which do not natively occur in the southern California region. creator: Matthew L. Gimmel creator: M. Andrew Johnston creator: Michael S. Caterino uri: https://doi.org/10.7717/peerj.14793 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Gimmel et al. title: Annual-scale assessment of mid-20th century anthropogenic impacts on the algal ecology of Crawford Lake, Ontario, Canada link: https://peerj.com/articles/14847 last-modified: 2023-03-08 description: Meromictic Crawford Lake, located in SW Ontario, Canada is characterized by varved sediments, making it suitable for high-resolution paleoecological studies. Freeze cores, the only coring method available that reliably preserves the fragile laminations representative of seasonal deposition in the lake, were used to document siliceous diatom and chrysophyte community structure at an annual resolution from 1930–1990CE. Stratigraphically constrained cluster analysis identified major assemblage changes that are believed to have been caused by local, regional and possibly global anthropogenic impacts. The assemblage changes within the siliceous algae are attributed to regional weather and increased industrial emissions and related effects of acid deposition on the lake’s catchment associated with the Great Acceleration –the massive economic, industrial, and demographic expansion beginning in the mid-20th century. Observed increases in spheroidal carbonaceous particles (SCPs) in varved lake sediment dating to the early 1950s record rapidly expanding steel production without emission controls around 30 km upwind of the lake. The findings reported here reflect major changes in earth systems that the Anthropocene Working Group recommends for a proposed epoch to be termed the Anthropocene, providing support for the laminated sediments from Crawford Lake as a potential Global boundary Stratotype Section and Point (GSSP). creator: Matthew G. Marshall creator: Paul B. Hamilton creator: Krysten M. Lafond creator: Nawaf A. Nasser creator: Francine M.G. McCarthy creator: R. Timothy Patterson uri: https://doi.org/10.7717/peerj.14847 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Marshall et al. title: Surviving without oxygen involves major tissue specific changes in the proteome of crucian carp (Carassius carassius) link: https://peerj.com/articles/14890 last-modified: 2023-03-08 description: The crucian carp (Carassius carassius) can survive complete oxygen depletion (anoxia) for several months at low temperatures, making it an excellent model for studying molecular adaptations to anoxia. Still, little is known about how its global proteome responds to anoxia and reoxygenation. By applying mass spectrometry-based proteome analyses on brain, heart and liver tissue from crucian carp exposed to normoxia, five days anoxia, and reoxygenation, we found major changes in particularly cardiac and hepatic protein levels in response to anoxia and reoxygenation. These included tissue-specific differences in mitochondrial proteins involved in aerobic respiration and mitochondrial membrane integrity. Enzymes in the electron transport system (ETS) decreased in heart and increased massively in liver during anoxia and reoxygenation but did not change in the brain. Importantly, the data support a special role for the liver in succinate handling upon reoxygenation, as suggested by a drastic increase of components of the ETS and uncoupling protein 2, which could allow for succinate metabolism without excessive formation of reactive oxygen species (ROS). Also during reoxygenation, the levels of proteins involved in the cristae junction organization of the mitochondria changed in the heart, possibly functioning to suppress ROS formation. Furthermore, proteins involved in immune (complement) system activation changed in the anoxic heart compared to normoxic controls. The results emphasize that responses to anoxia are highly tissue-specific and related to organ function. creator: Anette Johansen creator: Bernd Thiede creator: Jan Haug Anonsen creator: Göran E. Nilsson uri: https://doi.org/10.7717/peerj.14890 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Johansen et al. title: Rapid point-of-care detection of BK virus in urine by an HFman probe-based loop-mediated isothermal amplification assay and a finger-driven microfluidic chip link: https://peerj.com/articles/14943 last-modified: 2023-03-08 description: BackgroundBK virus (BKV)-associated nephropathy (BKVN) is one of the leading causes of renal dysfunction and graft loss in renal transplant recipients. Early monitoring of BKV in urine is crucial to minimize the deleterious effects caused by this virus on preservation of graft function.MethodsWe report a simple, rapid, sensitive loop-mediated isothermal amplification (LAMP) assay using an HFman probe for detecting BKV in urine. To evaluate the performance of the assay, a comparison of the HFman probe-based LAMP (HF-LAMP) assay with two qPCR assays was performed using urine samples from 132 HIV-1 infected individuals. We further evaluated the performance of HF-LAMP directly using the urine samples from these HIV-1 infected individuals and 30 kidney transplant recipients without DNA extraction. Furthermore, we combined the HF-LAMP assay with a portable finger-driven microfluidic chip for point-of-care testing (POCT).ResultsThe assay has high specificity and sensitivity with a limit of detection (LOD) of 12 copies/reaction and can be completed within 30 min. When the DNA was extracted, the HF-LAMP assay showed an equivalent and potentially even higher sensitivity (93.5%) than the qPCR assays (74.2–87.1%) for 132 urine samples from HIV-1 infected individuals. The HF-LAMP assay can be applied in an extraction-free format and can be completed within 45 min using a simple heat block. Although some decreased performance was seen on urine samples from HIV-1 infected individuals, the sensitivity, specificity, and accuracy of the extraction-free BKV HF-LAMP assay were 95%, 100%, and 96.7% for 30 clinical urine samples from kidney transplant recipients, respectively.ConclusionThe assay has high specificity and sensitivity. Combined with a portable finger-driven microfluidic chip for easy detection, this method shows great potential for POCT detection of BKV. creator: Yongjuan Zhao creator: Yi Zeng creator: Renfei Lu creator: Zhiying Wang creator: Xiaoling Zhang creator: Nannan Wu creator: Tongyu Zhu creator: Yang Wang creator: Chiyu Zhang uri: https://doi.org/10.7717/peerj.14943 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Zhao et al. title: Mezcal worm in a bottle: DNA evidence suggests a single moth species link: https://peerj.com/articles/14948 last-modified: 2023-03-08 description: Mezcals are distilled Mexican alcoholic beverages consumed by many people across the globe. One of the most popular mezcals is tequila, but there are other forms of mezcal whose production has been part of Mexican culture since the 17th century. It was not until the 1940–50s when the mezcal worm, also known as the “tequila worm”, was placed inside bottles of non-tequila mezcal before distribution. These bottled larvae increased public attention for mezcal, especially in Asia, Europe, and the United States. Despite these larvae gaining global interest, their identity has largely remained uncertain other than that they are larvae of one of three distantly related holometabolous insects. We sequenced the COI gene from larvae in different kinds of commercially available mezcals. All larval DNA that amplified was identified as the agave redworm moth, Comadia redtenbacheri. Those that did not amplify were also confirmed morphologically to be the larva of this species. creator: Akito Y. Kawahara creator: Jose I. Martinez creator: David Plotkin creator: Amanda Markee creator: Violet Butterwort creator: Christian D. Couch creator: Emmanuel F.A. Toussaint uri: https://doi.org/10.7717/peerj.14948 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Kawahara et al. title: New records and species of deep-sea squat lobsters (Galatheoidea, Munidopsidae) from the Hawaiian Archipelago: an integrative approach using micro-CT and barcodes link: https://peerj.com/articles/14956 last-modified: 2023-03-08 description: The Hawaiian Archipelago remains extensively under-sampled for many marine invertebrate taxa, including squat lobsters. During the last few years, several deep-sea expeditions carried out in the Pacific Ocean have conducted opportunistic collections of specimens and image data from the vicinity of Hawai’i. Here we describe a new species: Munidopsis hawaiisp. nov. and provide new records for Munidopsidae in the Archipelago and its associations. We illustrate and describe the new species using an integrative approach including micro-CT 3D imaging. Phylogenetic analyses of the species collected from seamounts from Hawai’i indicate that the new species represents a divergent lineage compared to morphologically similar species such as M. dispar and M. papanui. We also study the genetic distances for the species recorded in Hawai’i and other populations of the same species in the adjacent West Pacific. Three species are now known in the Hawaiian region. We also compiled identifications from images captured with ROVs in the area. These observations suggest that munidopsid species are common in the deep sea of Hawaiian waters below 1,000 m. creator: Paula C. Rodríguez Flores creator: Kareen E. Schnabel uri: https://doi.org/10.7717/peerj.14956 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Rodríguez Flores and Schnabel title: Genetic features and phylogenetic relationship analyses of Guizhou Han population residing in Southwest China via 38 X-InDels link: https://peerj.com/articles/14964 last-modified: 2023-03-08 description: BackgroundThe insertion/deletion polymorphism (InDel), an ideal forensic genetic marker with a low spontaneous mutation rate and small amplification product fragments, is widely distributed in the genome, combining the advantages of STR and SNP genetic markers. The X-chromosome has high application value in complex paternity testing, and it is an excellent system for evaluating population admixture and studying evolutionary anthropology. However, further research is needed on the population genetics of X-chromosome InDels (X-InDels).MethodsIn this article, a system composed of 38 X-InDel loci was utilized to analyse and evaluate the forensic parameters of the Guizhou Han population in order to explore its forensic application efficiency.ResultsThe results showed that expected heterozygosities spanned from 0.0189 to 0.5715, and the cumulative power of discrimination of the 32 X-InDels and three linkage blocks was 0.9999999954 and 0.999999999999741 for males and females, respectively. The combined mean exclusion chance of these loci for trios and duos is 0.999999 and 0.999747, respectively. Multiple methods like principal component analysis, Fst genetic distance, and phylogenetic reconstruction were employed for dissecting the genetic structure of the Guizhou Han population by comparing it with previously reported populations. As expected, the studied Han population displayed relatively close genetic affinities with the East Asian populations. At the same time, there were obvious genetic differentiations between the Guizhou Han population and other continental populations that were discerned, especially for the African populations.ConclusionsThis study further verified the applicability of 38 X-InDels for human personal identification and kinship analyses of Han Chinese, and also showed the application potential of X-InDels in population genetics. creator: Yuhang Feng creator: Ting Wang creator: Yunteng Yang creator: Jiangtao You creator: Kun He creator: Hongling Zhang creator: Qiyan Wang creator: Meiqing Yang creator: Jiang Huang creator: Zheng Ren creator: Xiaoye Jin uri: https://doi.org/10.7717/peerj.14964 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Feng et al. title: Transcriptome analysis and exploration of genes involved in the biosynthesis of secoiridoids in Gentiana rhodantha link: https://peerj.com/articles/14968 last-modified: 2023-03-08 description: Gentiana rhodantha is a medicinally important perennial herb used as traditional Chinese and ethnic medicines. Secoiridoids are one of the major bioactive compounds in G. rhodantha. To better understand the secoiridoid biosynthesis pathway, we generated transcriptome sequences from four organs (root, leaf, stem and flower), followed by the de novo sequence assembly. We verified 8-HGO (8-hydroxygeraniol oxidoreductase), which may encode key enzymes of the secoiridoid biosynthesis by qRT-PCR. The mangiferin, swertiamarin and loganic acid contents in root, stem, leaf, and flower were determined by HPLC. The results showed that there were 47,871 unigenes with an average length of 1,107.38 bp. Among them, 1,422 unigenes were involved in 25 standard secondary metabolism-related pathways in the KEGG database. Furthermore, we found that 1,005 unigenes can be divided into 66 transcription factor (TF) families, with no family members exhibiting significant organ-specificity. There were 54 unigenes in G. rhodantha that encoded 17 key enzymes of the secoiridoid biosynthetic pathway. The qRT-PCR of the 8-HGO and HPLC results showed that the relative expression and the mangiferin, swertiamarin, and loganic acid contents of the aerial parts were higher than in the root. Six types of SSR were identified by SSR analysis of unigenes: mono-nucleoside repeat SSR, di-nucleoside repeat SSR, tri-nucleoside repeat SSR, tetra-nucleoside repeat SSR, penta-nucleoside repeat SSR, and hexa-nucleoside repeat SSR. This report not only enriches the Gentiana transcriptome database but helps further study the function and regulation of active component biosynthesis of G. rhodantha. creator: Ting Zhang creator: Miaomiao Wang creator: Zhaoju Li creator: Xien Wu creator: Xiaoli Liu uri: https://doi.org/10.7717/peerj.14968 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Zhang et al. title: Two new species of Eleutherodactylus from western and central Mexico (Eleutherodactylus jamesdixoni sp. nov., Eleutherodactylus humboldti sp. nov.) link: https://peerj.com/articles/14985 last-modified: 2023-03-08 description: BackgroundThe subgenus Syrrhophus (genus Eleutherodactylus) contains >40 species of small, direct-developing frogs that occur at low to moderate elevations from Texas through Mexico and into Guatemala and Belize, with two species in western Cuba. Morphological conservatism and phenotypic convergence have made species delimitation challenging and resulted in a complicated taxonomic history. Since 2015, molecular systematic work has uncovered eleven new species from western Mexico and one from eastern Mexico, but current taxonomy still underestimates species level diversity and there is confusion surrounding the validity and boundary of several species.MethodsWe used phylogenetic analysis of 16S ribosomal RNA (rRNA) sequences, multivariate statistical analysis of morphological data, and bioacoustic analysis of male advertisement calls to discover two additional unnamed species of Eleutherodactylus from Central and Western Mexico. We describe those species here.ResultsEleutherodactylus (Syrrhophus) humboldti sp. nov. is described from the Quaternary Valle de Bravo volcanic field of the Eje Neovolcánico in Central Mexico. This species is sister to E. maurus and is 3% divergent in 16S. Eleutherodactylus (Syrrhophus) jamesdixoni sp. nov. is described from the Sierra Madre Occidental of western Mexico. This species is sister to E. nitidus and is 3% divergent. We provide color photographs, advertisement call recordings, and molecular diagnoses of these new species and their sister species to aid future workers. creator: Thomas J. Devitt creator: Karen Tseng creator: Marlena Taylor-Adair creator: Sannidhi Koganti creator: Alice Timugura creator: David C. Cannatella uri: https://doi.org/10.7717/peerj.14985 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Devitt et al. title: Dictyota defense: Developing effective chemical protection against intense fish predation for outplanted massive corals link: https://peerj.com/articles/14995 last-modified: 2023-03-08 description: The incorporation of coral species with massive (e.g., boulder, brain) morphologies into reef restoration is critical to sustain biodiversity and increase coral cover on degraded reef ecosystems. However, fragments and colonies of massive corals outplanted in Miami-Dade County, Florida, US, can experience intense predation by fish within the first week of outplanting, resulting in >70% mortality. Here, we tested for the first time the potential benefit of feeding corals powdered Dictyota, a brown reef alga that is chemically defended against grazing, to determine if exposure to Dictyota can confer chemical protection to coral fragments and reduce the impacts of fish predation after outplanting. We found that feeding corals every 2 to 3 days for 2 months with dried and powdered Dictyota prior to outplanting significantly reduced predation levels on Orbicella faveolata and Montastraea cavernosa fragments (with less than 20% of the fragments experiencing predation up to 1-month post-outplanting). We also found that a single exposure to Dictyota at a high concentration 1 to 2 days prior to outplanting significantly reduced predation for six coral species within the first 24 h following outplanting. Thus, feeding corals dry Dictyota ex situ prior to outplanting appears to confer protection from fish predation during the critical first days to weeks after outplanting when predation impacts are commonly high. This simple and cheap method can be easily scaled up for corals kept ex situ prior to outplanting, resulting in an increase in restoration efficiency for massive corals in areas with high fish predation. creator: Cailin Harrell creator: Diego Lirman uri: https://doi.org/10.7717/peerj.14995 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Harrell and Lirman title: Small worms of Sthenelanellinae, Pholoinae and Pisioninae (Annelida, Sigalionidae) from the Tropical Northwestern Atlantic link: https://peerj.com/articles/15005 last-modified: 2023-03-08 description: The family Sigalionidae is characterized, among other features, by including scale worms with large bodies. However, among sigalionids, the subfamilies Sthenelanellinae, Pholoinae, and Pisioninae stand out by their small representatives with few segments and fragile bodies. In the Tropical Northwestern Atlantic, which includes part of the Gulf of Mexico and the Caribbean Sea, these subfamilies have been rarely studied, with few species recorded, and questionable records. This contribution aims to improve the knowledge of sthenelanellins, pholoins, and pisionins in the region through a faunistic study based on material from two Mexican scientific collections: the Reference Collection of Laboratorio de Biodiversidad y Cambio Climático (BIOMARCCA) and the Reference Collection of Benthos (ECOSUR) of El Colegio de la Frontera Sur. Pisione wolfi is confirmed from the Tropical Northwestern Atlantic, and three new species are described: Sthenelanella pechi sp. nov., S. sarae sp. nov. and Taylorpholoe anabelae sp. nov. A key to all Sthenelanella and Taylorpholoe species is also included. creator: Christopher Cruz-Gómez uri: https://doi.org/10.7717/peerj.15005 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Cruz-Gómez title: Determinants of age-related decline in walking speed in older women link: https://peerj.com/articles/14728 last-modified: 2023-03-07 description: BackgroundWalking speed is reduced with aging. However, it is not certain whether the reduced walking speed is associated with physical and coordination fitness. This study explores the physical and coordination determinants of the walking speed decline in older women.MethodsOne-hundred-eighty-seven active older women (72.2 ± 6.8 years) were asked to perform a 10-m walk test (self-selected and maximal walking speed) and a battery of the Senior fitness test: lower body strength, lower body flexibility, agility/dynamic balance, and aerobic endurance. Two parameters characterized the walking performance: closeness to the modeled speed minimizing the energetic cost per unit distance (locomotor rehabilitation index, LRI), and the ratio of step length to step cadence (walk ratio, WR). For dependent variables (self-selected and maximal walking speeds), a recursive partitioning algorithm (classification and regression tree) was adopted, highlighting interactions across all the independent variables.ResultsParticipants were aged from 60 to 88 years, and their self-selected and maximal speeds declined by 22% and 26% (p < 0.05), respectively. Similarly, all physical fitness variables worsened with aging (muscle strength: 33%; flexibility: 0 to −8 cm; balance: 22%; aerobic endurance: 12%; all p < 0.050). The predictors of maximal walking speed were only WR and balance. No meaningful predictions could be made using LRI and WR as dependent variables.DiscussionThe results suggest that at self-selected speed, the decrease in speed itself is sufficient to compensate for the age-related decline in the motor functions tested; by contrast, lowering the WR is required at maximal speed, presumably to prevent imbalance. Therefore, any excessive lowering of LRI and WR indicates loss of homeostasis of walking mechanics and invites diagnostic investigation. creator: Valéria Feijó Martins creator: Luigi Tesio creator: Anna Simone creator: Andréa Kruger Gonçalves creator: Leonardo A. Peyré-Tartaruga uri: https://doi.org/10.7717/peerj.14728 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Martins et al. title: Quantitative trait loci associated with straighthead-resistance used for marker assisted selection in rice (Oryza sativa L.) RIL populations link: https://peerj.com/articles/14866 last-modified: 2023-03-07 description: Straighthead is a physiological disorder of rice (Oryza sativa L.) that causes dramatic yield loss in susceptible cultivars. This disorder is found worldwide and is reported to increasingly occur in the southern United States. Genetic resistance breeding has been considered as one of the most efficient methods for straighthead prevention because the traditional prevention method wastes water and costs labor. In this study, we analyzed the genetic effects of five straighthead quantitative trait loci (QTLs), namely, AP3858-1 (qSH-8), RM225 (qSH-6), RM2 (qSH-7), RM206 (qSH-11), and RM282 (qSH-3), on the recombinant inbred lines (RILs) developed from Jing185/Cocodrie and Zhe733/R312 populations using our five previously identified markers linked to these QTLs. As a result, recombinant inbred lines (RILs) with four resistant alleles at the four loci (AP3858-1, RM225, RM2, and RM206) exhibited the highest straighthead resistance. This result suggests that the four markers could be efficiently used to select the straighthead-resistant recombinant inbred lines (RILs). Furthermore, by using AP3858-1, we successfully obtained five straighthead-resistant recombinant inbred lines (RILs) with more than 50% genetic similarity to Cocodrie. These markers and recombinant inbred lines (RILs) can be used for future straighthead resistance breeding through marker-assisted selection. creator: Xuhao Pan creator: Yiting Li creator: Xiaobai Li uri: https://doi.org/10.7717/peerj.14866 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Pan et al. title: RNA-seq reveals Nup62 as a potential regulator for cell division after traumatic brain injury in mice hippocampus link: https://peerj.com/articles/14913 last-modified: 2023-03-07 description: BackgroundHippocampus impairment is a common condition encountered in the clinical diagnosis and treatment of traumatic brain injury (TBI). Several studies have investigated this phenomenon. However, its molecular mechanism remains unclear.MethodsIn this study, Illumina RNA-seq technology was used to determine the gene expression profile in mice hippocampus after TBI. We then conducted bioinformatics analysis to identify the altered gene expression signatures and mechanisms related to TBI-induced pathology in the hippocampus. Real-time quantitative polymerase chain reaction and western blot were adopted to verify the sequencing results.ResultsThe controlled cortical impact was adopted as the TBI model. Hippocampal specimens were removed for sequencing. Bioinformatics analysis identified 27 upregulated and 17 downregulated differentially expressed genes (DEGs) in post-TBI mouse models. Potential biological functions of the genes were determined via Gene Set Enrichment Analysis (GSEA)-based Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, which suggested a series of functional changes in the nervous system. Specifically, the nucleoporin 62 (Nup62) DEG was discussed and verified. Gene ontology biological process enriched analysis suggests that the cell division was upregulated significantly. The present study may be helpful for the treatment of impaired hippocampus after TBI in the future. creator: Jianwei Zhao creator: Weihua Wang creator: Ke Yan creator: Haifeng Zhao creator: Zhen Zhang creator: Yu Wang creator: Wenyu Zhu creator: Shiwen Chen uri: https://doi.org/10.7717/peerj.14913 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Zhao et al. title: Functional verification of the JmLFY gene associated with the flowering of Juglans mandshurica Maxim. link: https://peerj.com/articles/14938 last-modified: 2023-03-07 description: In this study, a pBI121-JmLFY plant expression vector was constructed on the basis of obtaining the full-length sequence of the JmLFY gene from Juglans mandshurica, which was then used for genetic transformation via Agrobacterium inflorescence infection using wild-type Arabidopsis thaliana and lfy mutants as transgenic receptors. Seeds of positive A. thaliana plants with high expression of JmLFY were collected and sowed till the homozygous T3 regeneration plants were obtained. Then the expression of flowering-related genes (AtAP1, AtSOC1, AtFT and AtPI) in T3 generation plants were analyzed and the results showed that JmLFY gene overexpression promoted the expression of flowering-related genes and resulted in earlier flowering in A. thaliana. The A. thaliana plants of JmLFY-transformed and JmLFY-transformed lfy mutants appeared shorter leaves, longer fruit pods, and fewer cauline leaves than those of wild-type and the lfy mutants plants, respectively. In addition, some secondary branches in the transgenic plants converted into inflorescences, which indicated that the overexpression of JmLFY promoted the transition from vegetative growth to reproductive growth, and compensate the phenotypic defects of lfy mutant partially. The results provides a scientific reference for formulating reasonable genetic improvement strategies such as shortening childhood, improving yield and quality, and breeding desirable varieties, which have important guiding significance in production. creator: Jiayou Cai creator: Ruoxue Jia creator: Ying Jiang creator: Jingqi Fu creator: Tianyi Dong creator: Jifeng Deng creator: Lijie Zhang uri: https://doi.org/10.7717/peerj.14938 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Cai et al. title: Apocynum venetum, a medicinal, economical and ecological plant: a review update link: https://peerj.com/articles/14966 last-modified: 2023-03-07 description: Apocynum venetum L. is an important medicinal perennial rhizome plant with good ecological and economic value. Its leaves have many pharmacological effects such as anti-inflammatory, anti-depression, anti-anxiolytic, etc., while its fibers have the title of “king of wild fibers”. Furthermore, it was suitable for the restoration of degraded saline soil in arid areas. An increasing studies have been published in the past years. A scientometric analysis was used to analyze the publications of Apocynum venetum L. to clearly review the pharmacology, fiber application of Apocynum venetum L. and the potential value with its similar species (Apocynum pictum Schrenk) to the environment. creator: Tian Xiang creator: Longjiang Wu creator: Murtala Bindawa Isah creator: Chen Chen creator: Xiaoying Zhang uri: https://doi.org/10.7717/peerj.14966 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Xiang et al. title: Uncertainties regarding the natural mortality of fish can increase due global climate change link: https://peerj.com/articles/14989 last-modified: 2023-03-07 description: The increase in temperature resulting from global climate change can directly affect the survival of fish and therefore population parameters such as natural mortality (M). The estimation of this parameter and the understanding of the uncertainties in its estimates are enormous challenges for studies that evaluate fish stocks. In addition, the effects of increases in temperature may be associated with life strategies. Therefore, the fuzzy set theory was used to evaluate the effects of temperature increase on the natural mortality of fish, considering different life strategies. The model showed that the increase in temperature increased the uncertainties in M estimates for all species, regardless of the life strategy. However, opportunistic species present greater uncertainties in estimates of M compared to equilibrium species. The patterns found in uncertainties of M associated with species groupings by life strategies can be used in holistic approaches for the assessment and management of recently exploited fisheries resources or for those with limited biological data. creator: Caroline Pereira Campos creator: Sandro Dimy Barbosa Bitar creator: Carlos Freitas uri: https://doi.org/10.7717/peerj.14989 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Pereira Campos et al. title: Comparative ultrastructure of the antennae and sensory hairs in six species of crayfish link: https://peerj.com/articles/15006 last-modified: 2023-03-07 description: BackgroundAntennae in crayfish are essential for gaining information about the local topography and localising food, chemicals, conspecifics or predator. There are still gaps in the research on the morphology of antennae in decapods compared to other arthropods.MethodologyBiometrical and ultrastructural methods were applied using light and cryo-scanning electron microscopies to study the morphology of antennae in six different crayfish species, including marbled crayfish Procambarus virginalis, Mexican dwarf crayfish Cambarellus patzcuarensis, red swamp crayfish Procambarus clarkii, signal crayfish Pacifastacus leniusculus, common yabby Cherax destructor, and spiny-cheek crayfish Faxonius limosus to find their potential morphological differences.ResultsSignificant differences in the antenna length, length and width of each segment to carapace length ratios, and the number of segments were found in the six crayfish species. The ultrastructure revealed differences in the distribution of sensory hairs on the antenna and the morphology of the antennal surface.ConclusionsThe different morphology of antennae might reflect adaptation to the conditions of their specific habitats. In addition, results showed that a combination of differences in the morphological features and biometrical measurements of antennae could be used for the distinguishment of different studied crayfish species. creator: Golara Kor creator: Kifayatullah Mengal creator: Miloš Buřič creator: Pavel Kozák creator: Hamid Niksirat uri: https://doi.org/10.7717/peerj.15006 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Kor et al. title: The study of the characteristics of the secondary flowering of Cerasus subhirtella ‘Autumnalis’ link: https://peerj.com/articles/14655 last-modified: 2023-03-06 description: The short flowering period of ornamental cherry trees is the main factor limiting their use in gardens. Determining the secondary flowering characteristics of ornamental cherry trees is required to prolong their flowering period. In this experiment, Cerasus subhirtella ‘Autumnalis’ was used as the experimental material. The phenological differences in their annual growth cycle were observed using the BBCH coding system. The cooling requirements of the flower buds were evaluated by the chilling hours model (temperature between 0 and 7.2 °C) and the Utah model. The expression of the core gene involved in bud dormancy regulation DAM (dormancy-associated MADS-box) from the completion of flower bud differentiation in one year until the following year was measured by performing real-time fluorescence-based quantitative PCR. The results showed that the flowering duration of C. subhirtella ‘Autumnalis’ from November to December was longer than that of C. yedoensis ‘Somei Yoshino’, which was from March to April. The progress from seed bud-break to flower bud opening took about 10 days for C. subhirtella ‘Autumnalis’, while the same stage for C. yedoensis ‘Somei Yoshino’ took around 20 days. Additionally, the flower buds of C. subhirtella ‘Autumnalis’ needed only the chilling temperature unit of 54.08 to satisfy the chilling requirement, while C. yedoensis ‘Somei Yoshino’ required a chilling temperature unit of 596.75. After the completion of flower bud differentiation, during low-temperature storage, the expression of DAM4 and DAM5 genes first increased and then decreased, whereas, the expression of the DAM6 gene continued to decrease, and the expression of DAM4, DAM5, and DAM6 in C. yedoensis ‘Somei Yoshino’ increased rapidly and was maintained at a high level. This showed that the upregulation of the expression of the DAM4, DAM5, and DAM6 genes can inhibit the flower bud germination of Cherry Blossom. The relative expression of the DAM gene of C. subhirtella ‘Autumnalis’ was significantly lower than that of the DAM gene of C. yedoensis ‘Somei Yoshino’ from the end of October to the beginning of December, leading to its secondary flowering in autumn. These results might elucidate why the flower buds of C. subhirtella ‘Autumnalis’ can break their internal dormancy and bloom in the autumn and then again in the following year. Our findings might provide a reference for conducting further studies on the mechanisms of secondary flowering and bud dormancy in cherries. creator: Yanxia Xu creator: Liyan Du creator: Xuebin Song creator: Chunling Zhou uri: https://doi.org/10.7717/peerj.14655 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Xu et al. title: Microbial diversity and abundance in loamy sandy soil under renaturalization of former arable land link: https://peerj.com/articles/14761 last-modified: 2023-03-06 description: The abundance and taxonomic diversity of different physiological groups of bacteria and fungi and yeasts in the fields of the long-term experiment of renaturalization of infertile arable soils were studied. The experiment involved four land conversion methods: conversion of arable land to cultivated meadow, soil and forest, leaving the experimental area of arable land. With these studies, we have begun to fill research gaps related to the taxonomic and functional diversity of soil microorganisms. The greatest changes in the abundance of cultivable organotrophic, diazotrophic and nitrifying bacteria were found to be observed in those areas where anthropogenic activities took place, i.e. in the cultivated field and in the cultural grassland. The abundance of bacteria was relatively lower and that of fungi was higher in the soil and in the cultivated area. It was also found that the higher jumps in the abundance of diazotrophs and nitrifiers during the respective stages of vegetation were caused by the applied agrotechnical measures and the cultivation of the respective plants. The abundance of cultivable bacteria was up to 105, and the number of fungi was 103 CFU in 1 g of dry soil. The taxonomic structure was determined by Next Generation Sequencing. The taxonomic groups of Actino- and Proteobacteria had the highest abundance. The highest number of fungal OTU was distinguished by Ascomycota fungi (37–42% of the total number of fungi). Comparing the taxonomic structure of all studied samples, the area planted with pines stands out, where an increase in the taxonomic group of Basidiomycota fungi (up to 24%) is observed at the expense of Ascomycota fungi. In order to have a balanced, fully rich soil, efforts must be made to maintain a stable structure of microbial communities, which can only be achieved through targeted research. creator: Audrius Kacergius creator: Diana Sivojiene uri: https://doi.org/10.7717/peerj.14761 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Kacergius and Sivojiene title: Oral health related quality of life and the prevalence of ageusia and xerostomia in active and recovered COVID-19 Patients link: https://peerj.com/articles/14860 last-modified: 2023-03-06 description: BackgroundSalivary disturbance is associated with patients who either have an active coronavirus disease 2019 (COVID-19) or have recovered from coronavirus infection along with loss of taste sensation. In addition, COVID-19 infection can drastically compromise quality of life of individuals.ObjectiveThis study aimed to analyze xerostomia, ageusia and the oral health impact in coronavirus disease-19 patients utilizing the Xerostomia Inventory scale-(XI) and the Oral Health Impact Profile-14.MethodsIn this cross-sectional survey-based study, data was collected from 301 patients who suffered and recovered from COVID-19. Using Google Forms, a questionnaire was developed and circulated amongst those who were infected and recovered from coronavirus infection. The Xerostomia Inventory (XI) and Oral Health Impact Profile-14 were used to assess the degree and quality of life. A paired T-test and Chi-square test were used to analyze the effect on xerostomia inventory scale-(XI) and OHIP-14 scale scores. A p-value of 0.05 was considered as statistically significant.ResultsAmong 301 participants, 54.8% were females. The prevalence of xerostomia in participants with active COVID-19 disease was 39.53% and after recovery 34.88%. The total OHIP-14 scores for patients in the active phase of infection was 12.09, while 12.68 in recovered patients. A significant difference was found between the mean scores of the xerostomia inventory scale-11 and OHIP-14 in active and recovered COVID patients.ConclusionA higher prevalence of xerostomia was found in COVID-19 infected patients (39.53%) compared to recovered patients (34.88%). In addition, more than 70% reported aguesia. COVID-19 had a significantly higher compromising impact on oral function of active infected patients compared to recovered patients. creator: Mahnoor K.M. Saleem creator: Abhishek Lal creator: Naseer Ahmed creator: Maria S. Abbasi creator: Fahim Vohra creator: Tariq Abduljabbar uri: https://doi.org/10.7717/peerj.14860 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Saleem et al. title: METTL3-mediated m6A modification of has_circ_0007905 promotes age-related cataract progression through miR-6749-3p/EIF4EBP1 link: https://peerj.com/articles/14863 last-modified: 2023-03-06 description: Many cases of blindness are caused by age-related cataracts (ARCs). N6-methyladenosine (m6A)-modified circRNA widely participates in disease progression. However, the role of m6A modification of circRNA in ARC is unclear. We mined and elucidated the functions and mechanisms of key circRNAs with m6A modification involved in ARC progression. The GSE153722 dataset was used to mine m6A-mediated key circRNA. Loss-of-function assays and rescue assays were used to explore the effect and mechanism of circRNA on ARC cell proliferation and apoptosis. Has_circ_0007905 was a hypermethylated and upregulated expression in the ARC group relative to the control group both in vivo and in vitro. Silencing of has_circ_0007905 promoted proliferation and inhibited the apoptosis of HLE-B3 cells. METTL3 was upregulated in HLE-B3 cells after ARC modeling and had four binding sites with has_circ_0007905 and a mediated m6A modification of has_circ_0007905. Proliferation was significantly inhibited and apoptosis of HLE-B3 cells was facilitated by METTL3 overexpression, whereas these effects were prevented by has_circ_0007905 silencing. Silencing of has_circ_0007905 led to an alteration in the transcriptome landscape. Differentially expressed genes were mainly involved in immune-related processes and pathways. EIF4EBP1 overexpression promoted apoptosis and suppressed proliferation, and also significantly reversed effects of has_circ_0007905 silencing. Moreover, miR-6749-3p significantly decreased the luciferase activities of wild type plasmids with both of has_circ_0007905 and EIF4EBP1. MiR-6749-3p inhibitor blocked elevation in proliferation and reduced EIF4EBP1 expression and apoptosis conferred by has_circ_0007905 silencing. We reveal for the first time that the commitment of ARC progression is guided by METTL3/has_circ_0007905/miR-6749-3p/EIF4EBP1 axis, and the results provide new insights into ARC pathology. creator: Rui Li creator: Haohao Zhu creator: Qian Li creator: Jiancen Tang creator: Yiping Jin creator: Hongping Cui uri: https://doi.org/10.7717/peerj.14863 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Li et al. title: The first record of chigutisaurid amphibian from the Late Triassic Tiki Formation and the probable Carnian pluvial episode in central India link: https://peerj.com/articles/14865 last-modified: 2023-03-06 description: A new, partially preserved skull of chigutisaurid amphibian (temnospondyli) has been reported for the first time from the Late Triassic Tiki Formation of India. Chigutisaurids are now known to occur in Australia’s Early and Late Triassic, the Late Triassic in India, Argentina, and Brazil, the Jurassic of South Africa and Australia, and the Cretaceous of Australia. In India, the first appearance of chigutisaurids marks the Carnian—middle Carnian/Norian Boundary. This work also attempts to correlate, again for the first time, the advent of chigutisaurids and the occurrence of Carnian Pluvial Episodes (CPE) in the Late Triassic Maleri and Tiki Formation of Central India. The new specimen belongs to the genus Compsocerops prevalent in the Late Triassic Maleri Formation occurring 700 km south. However, the chigutisaurid specimen recovered from the Tiki Formation is a new species when compared to that of the Maleri Formation. It has the presence of an inward curved process of the quadratojugal as opposed to the straight downward trending process of the quadratojugal, the presence of vomerine foramen, shorter and wider interpterygoid vacuities, wider subtemporal vacuities, and the base of the interpterygoid vacuities at the same level with the base of the subtemporal vacuity. It proves that the Tiki Formation is coeval with the Lower Maleri Formation and a part of Upper Maleri. creator: Sanjukta Chakravorti creator: Dhurjati Prasad Sengupta uri: https://doi.org/10.7717/peerj.14865 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Chakravorti et al. title: Six new species of free-living nematodes (Nematoda: Enoplida) from deep-sea cold seeps on Hikurangi Margin, New Zealand link: https://peerj.com/articles/14867 last-modified: 2023-03-06 description: Little is known about the taxonomy of deep-sea nematode species inhabiting cold seep habitats. An opportunity to characterize the nematode species communities of New Zealand cold seeps was provided by a 2019 research voyage to New Zealand’s Hikurangi Margin, during which macrofauna cores were obtained at two seeps at approximately 1,250 and 2,000 m water depth. Here, six new species of the orderEnoplida are described. Metacylicolaimus catherinae sp. nov. represents the first record of the genus for the New Zealand Exclusive Economic Zone and for the deep sea globally. Halalaimus talaurinus sp. nov., Thalassoalaimus duoporus sp. nov. and Crenopharynx crassipapilla sp. nov. are only the second species of their respective genera to be described/recorded from New Zealand waters, and Oncholaimus adustus sp. nov. is the eighth species of the genus to be recorded from the region. Rhabdodemania zealandiaensis sp. nov. was among the most abundant and widespread species found at the Hikurangi Margin seep sites. A few specimens had been found in a previous ecological study of meiofaunal nematode communities on Chatham Rise, a submarine ridge south of Hikurangi Margin. It is possible that this species has a preference for seep environments due to elevated food availability, however it does not seem to be exclusively found in seeps. We find no evidence for an affinity between nematode seep communities in New Zealand and elsewhere, which is consistent with the high variability in nematode community observed to date among regions. Ongoing work on the ecology and distribution of nematode communities at the Hikurangi Margin seep sites will help determine spatial patterns in abundance and species distributions in more detail, including the identification of any species/taxa with affinities with seeps. creator: Daniel Leduc uri: https://doi.org/10.7717/peerj.14867 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Leduc title: Abnormal mechanical stress on bicuspid aortic valve induces valvular calcification and inhibits Notch1/NICD/Runx2 signal link: https://peerj.com/articles/14950 last-modified: 2023-03-06 description: BackgroundBicuspid aortic valve (BAV) is a congenital cardiac deformity, increasing the risk of developing calcific aortic valve disease (CAVD). The disturbance of hemodynamics can induce valvular calcification, but the mechanism has not been fully identified.MethodsWe constructed a finite element model (FEM) of the aortic valve based on the computed tomography angiography (CTA) data from BAV patients and tricuspid aortic valve (TAV) individuals. We analyzed the hemodynamic properties based on our model and investigated the characteristics of mechanical stimuli on BAV. Further, we detected the expression of Notch, NICD and Runx2 in valve samples and identified the association between mechanical stress and the Notch1 signaling pathway.ResultsFinite element analysis showed that at diastole phase, the equivalent stress on the root of BAV was significantly higher than that on the TAV leaflet. Correspondingly, the expression of Notch1 and NICH decreased and the expression of Runx2 elevated significantly on large BAV leaflet belly, which is associated with equivalent stress on leaflet. Our findings indicated that the root of BAV suffered higher mechanical stress due to the abnormal hemodynamic environment, and the disturbance of the Notch1/NICD/Runx2 signaling pathway caused by mechanical stimuli contributed to valvular calcification. creator: Guangzhou Li creator: Na Shen creator: Huifang Deng creator: Yixuan Wang creator: Gangcheng Kong creator: Jiawei Shi creator: Nianguo Dong creator: Cheng Deng uri: https://doi.org/10.7717/peerj.14950 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Li et al. title: Reducing options of ammonia volatilization and improving nitrogen use efficiency via organic and inorganic amendments in wheat (Triticum aestivum L.) link: https://peerj.com/articles/14965 last-modified: 2023-03-06 description: BackgroundThis study investigates the effect of organic and inorganic supplements on the reduction of ammonia (NH3) volatilization, improvement in nitrogen use efficiency (NUE), and wheat yield.MethodsA field experiment was conducted following a randomized block design with 10 treatments i.e., T1-without nitrogen (control), T2-recommended dose of nitrogen (RDN), T3-(N-(n-butyl) thiophosphoric triamide) (NBPT @ 0.5% w/w of RDN), T4-hydroquinone (HQ @ 0.3% w/w of RDN), T5-calcium carbide (CaC2 @ 1% w/w of RDN), T6-vesicular arbuscular mycorrhiza (VAM @ 10 kg ha−1), T7-(azotobacter @ 50 g kg−1 seeds), T8-(garlic powder @ 0.8% w/w of RDN), T9-(linseed oil @ 0.06% w/w of RDN), T10-(pongamia oil @ 0.06% w/w of RDN).ResultsThe highest NH3 volatilization losses were observed in T2 at about 20.4 kg ha−1 per season. Significant reduction in NH3 volatilization losses were observed in T3 by 40%, T4 by 27%, and T8 by 17% when compared to the control treatment. Soil urease activity was found to be decreased in plots receiving amendments, T3, T4, and T5. The highest grain yield was observed in the T7 treated plot with 5.09 t ha−1, and straw yield of 9.44 t ha−1 in T4.ConclusionThe shifting towards organic amendments is a feasible option to reduce NH3 volatilization from wheat cultivation and improves NUE. creator: Pooja L. Ramalingappa creator: Manoj Shrivastava creator: Shiva Dhar creator: Kalikinkar Bandyopadhyay creator: Shiv Prasad creator: Sapna Langyan creator: Ritu Tomer creator: Ashish Khandelwal creator: Sibananda Darjee creator: Renu Singh uri: https://doi.org/10.7717/peerj.14965 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 L Ramalingappa et al. title: Community differentiation of rhizosphere microorganisms and their responses to environmental factors at different development stages of medicinal plant Glehnia littoralis link: https://peerj.com/articles/14988 last-modified: 2023-03-06 description: Rhizosphere microorganisms play a key role in affecting plant quality and productivity through its interaction with plant root system. To figure out the bottleneck of the decline of yield and quality in the traditional Chinese medicinal herbs Glehnia littoralis they now encounter, it is important to study the dynamics of rhizosphere microbiota during the cultivation of G. littoralis. In the present study, the composition, diversity and function of rhizosphere microbes at different development stages of G. littoralis, as well as the correlation between rhizosphere microbes and environmental factors were systematically studied by high-throughput sequencing. There were significant differences between the rhizosphere microbes at early and middle-late development stages. More beneficial bacteria, such as Proteobacteria, and more symbiotic and saprophytic fungi were observed at the middle-late development stage of G. littoralis, while beneficial bacteria such as Actinobacteria and polytrophic transitional fungi were abundant at all development stages. The results of redundancy analysis show that eight environmental factors drive the changes of microflora at different development stages. pH, soil organic matter (SOM) and available phosphorus (AP) had important positive effects on the bacterial and fungal communities at the early development stage; saccharase (SC) and nitrate nitrogen (NN) showed significant positive effects on the bacterial and fungal communities at the middle and late stages; while urease (UE), available potassium (AK), and alkaline phosphatase (AKP) have different effects on bacterial and fungal communities at different development stages. Random forest analysis identified 47 bacterial markers and 22 fungal markers that could be used to distinguish G. littoralis at different development stages. Network analysis showed that the rhizosphere microbes formed a complex mutualistic symbiosis network, which is beneficial to the growth and development of G. littoralis. These results suggest that host development stage and environmental factors have profound influence on the composition, diversity, community structure and function of plant rhizosphere microorganisms. This study provides a reference for optimizing the cultivation of G. littoralis. creator: Shuliang Liu creator: Jianxin Gao creator: Shimeng Wang creator: Weiwei Li creator: Ailan Wang uri: https://doi.org/10.7717/peerj.14988 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Liu et al. title: Genome-wide survey of HMA gene family and its characterization in wheat (Triticum aestivum) link: https://peerj.com/articles/14920 last-modified: 2023-03-03 description: BackgroundAbiotic stresses, particularly drought and heavy metal toxicity, have presented a significant risk to long-term agricultural output around the world. Although the heavy-metal-associated domain (HMA) gene family has been widely explored in Arabidopsis and other plants, it has not been thoroughly studied in wheat (Triticum aestivum). This study was proposed to investigate the HMA gene family in wheat.MethodsTo analyze the phylogenetic relationships, gene structure, gene ontology, and conserved motifs, a comparative study of wheat HMA genes with the Arabidopsis genome was performed.ResultsA total of 27 T. aestivum proteins belonging to the HMA gene family were identified in this study, with amino acid counts ranging from 262 to 1,071. HMA proteins were found to be grouped into three subgroups in a phylogenetic tree, and closely related proteins in the tree showed the same expression patterns as motifs found in distinct subgroups. Gene structural study elucidated that intron and exon arrangement differed by family.ConclusionAs a result, the current work offered important information regarding HMA family genes in the T. aestivum genome, which will be valuable in understanding their putative functions in other wheat species. creator: Sadaf Zahra creator: Tayyaba Shaheen creator: Muhammad Qasim creator: Mahmood-ur-Rahman creator: Momina Hussain creator: Sana Zulfiqar creator: Kanval Shaukat creator: Mehboob-ur-Rahman uri: https://doi.org/10.7717/peerj.14920 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Zahra et al. title: Prevalence and risk factors of computer vision syndrome—assessed in office workers by a validated questionnaire link: https://peerj.com/articles/14937 last-modified: 2023-03-03 description: BackgroundComputer vision syndrome (CVS) is a common occupational health problem, but its clinical definition, prevalence and risk factors are not well defined. In general, non-validated diagnostic instruments have been used to assess its prevalence. For this reason, the aim of this study is to estimate the prevalence and potential risk factors for CVS using a validated questionnaire.MethodsA cross-sectional study (n = 238) was carried out in Italian office workers using digital devices. All participants responded to an anamnesis, a digital exposure questionnaire, and the validated Italian version of the Computer Vision Syndrome Questionnaire. A battery of 3 ocular surface and tear ophthalmic tests (break-up time, BUT), Schirmer II and corneal staining) was performed.ResultsThe mean age (±SD) was 45.55 (11.02) years, 64.3% were female. 71.4% wore glasses to work, whose design was monofocal (for distance) in 47.6%, monofocal (for near) in 26.5%, general progressive in 16.5% and occupational progressive in 8.8% of cases. 35.7% used digital devices >6 hours/day in the workplace. The prevalence of CVS was 67.2%. In the multivariate model, female sex (aOR: 3.17; 95% CI [1.75–5.73]), the use of digital devices >6 hours/day at workplace (aOR: 2.07; 95% CI [1.09–3.95]) and the use of optical correction at work (aOR: 2.69; 95% CI [1.43–5.08]) significantly increased the odds of CVS. Association was observed between presenting CVS and having abnormal BUT (χ2 = 0.017).ConclusionsThe prevalence of CVS in Italian office workers, especially among females, was high. Intensive use of digital devices at work (>6 hours/day) and the use of optical correction at work significantly increased the odds of CVS. There is an association between poor tear stability and CVS. Further research is needed on the influence of wearing optical correction on CVS. The use of a validated questionnaire in health surveillance of digital workers is strongly recommended. creator: Natalia Cantó-Sancho creator: Stefano Porru creator: Stefano Casati creator: Elena Ronda creator: Mar Seguí-Crespo creator: Angela Carta uri: https://doi.org/10.7717/peerj.14937 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Cantó-Sancho et al. title: MicroRNA expression in apical periodontitis and pulpal inflammation: a systematic review link: https://peerj.com/articles/14949 last-modified: 2023-03-03 description: BackgroundThe aim of this systematic review is to determine microRNAs (miRs) that are differently expressed between diseased pulpal and periapical tissues.DesignThis systematic review used PubMed, Scopus, EBSCO, ProQuest, Cochrane database as well as manual searching to extract studies from January 2012 up to February 2022.ResultsA total of 12 studies met the eligibility criteria were included. All selected studies were of case-control type. Twenty-four miRNAs associated with apical periodontitis, 11 were found to be upregulatedand 13 were downregulated. Four out of the 44 miRs associated with pulpal inflammation were upregulated, whereas forty were downregulated. Six miRs, namely hsa-miR-181b, hsa-miR-181c,hsa-miR-455-3p,hsa-miR-128-3p, hsa-miR199a-5p, and hsa-miR-95, exhibited considerable downregulation in both periapical and pulp tissues.ConclusionMiRs have been investigated for their role in pulpal and periapical biology and may be utilised in diagnostic and therapeutic purposes. Further investigations are required to determine why certain irreversible pulpitis situations progress to apical periodontitis and others do not, based on the various miR expressions. Moreover, clinical and laboratory trials are needed to support this theory. creator: Zainab Jamal Al Gashaamy creator: Tiba Alomar creator: Linah Al-Sinjary creator: Mohammad Wazzan creator: Musab Hamed Saeed creator: Natheer H. Al-Rawi uri: https://doi.org/10.7717/peerj.14949 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Al Gashaamy et al. title: Long-term outcomes and predictors of patients with ST elevated versus non-ST elevated myocardial infarctions in non-obstructive coronary arteries: a retrospective study in Northern China link: https://peerj.com/articles/14958 last-modified: 2023-03-03 description: BackgroundMyocardial infarction with non-obstructive coronary arteries (MINOCA) is a heterogeneous disease entity with diverse etiologies and no uniform treatment protocols. Patients with MINOCA can be clinically classified into two groups based on whether they have an ST-segment elevation (STE) or non-ST segment elevation (NSTE), based on electrocardiogram (ECG) results, whose clinical prognosis is unclear. This study aimed to compare the outcomes and predictors of patients with STE and NSTE in the MINOCA population.MethodsWe collected the data for 196 patients with MINOCA (115 with STE and 81 with NSTE) in China. Clinical characteristics, prognoses, and predictors of major adverse cardiovascular events (MACE) were analyzed during the follow-up of all patients.ResultsThe proportion of patients with STE was greater than that with NSTE in the MINOCA population. Patients with NSTE were older and had a higher incidence of hypertension. No differences were observed in the outcomes between the STE and NSTE groups during a median follow-up period of 49 (37,46) months. No significant differences were observed in those with MACE (24.35% vs 22.22%, P = 0.73) and those without MACE. The multivariable predictors of MACE in the NSTE groups were Killip grades ≥ 2 (HR 9.035, CI 95% [1.657–49.263], P = 0.011), reduced use of β-blockers during hospitalization (HR 0.238, CI 95% [0.072–0.788], P = 0.019), and higher levels of low-density lipoprotein cholesterol (LDL-C) (HR 2.267, CI 95% [1.008–5.097], P = 0.048); the reduced use of β-blockers during hospitalization was the only independent risk factor of MACE in the STE group.ConclusionsThere were differences between the clinical characteristics of patients with STE and NSTE in the MINOCA population, even though outcomes during follow-up were similar. Independent risk factors for major adverse cardiac events were not identical in the STE and NSTE groups, which could be attributable to the differences in disease pathogenesis. creator: Lin Chen creator: Yinghong Fan creator: Zhen Fang creator: Ning Liu uri: https://doi.org/10.7717/peerj.14958 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Chen et al. title: CYP27A1 deficiency promoted osteoclast differentiation link: https://peerj.com/articles/15041 last-modified: 2023-03-03 description: BackgroundThe elevating osteoclast differentiation can lead to an imbalance in bone homeostasis, which was responsible for bone loss and bone diseases, such as osteoporosis. Multiple pathways and molecules have been involved in osteoclast formation, but the role of CYP27A1 in osteoclast differentiation has never been explored.MethodsCYP27A1 deficient mice were constructed using CRISPR-Cas9 system. Osteoclast differentiation was detected by TRAP staining. Differentially expressed genes (DEGs) were identified using RNA-seq analysis and were confirmed by qRT-PCR and Western blot.ResultsThe results showed that CYP27A1 knockout (KO) promoted osteoclast differentiation and bone loss. The transcriptomic analysis revealed that CYP27A1 KO led to differential expression of multiple genes, including ELANE, LY6C2, S100A9, GM20708, BGN, SPARC, and COL1A2, which were confirmed by qRT-PCR and Western blot. Enrichment analysis indicated that these differential genes were significantly associated with osteogenesis-related pathways, such as PPAR signaling, IL-17 signaling, and PI3K/AKT signaling, which were confirmed by qRT-PCR and Western blot.ConclusionsThese results suggested that CYP27A1 was involved in osteoclast differentiation, providing a novel therapeutic target for osteoclast-related diseases. creator: Ziqi Fang creator: Guangdong Cheng creator: Mengting He creator: Yanliang Lin uri: https://doi.org/10.7717/peerj.15041 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Fang et al. title: Adaptive mechanisms in quinoa for coping in stressful environments: an update link: https://peerj.com/articles/14832 last-modified: 2023-03-02 description: Quinoa (Chenopodium quinoa) is a grain-like, genetically diverse, highly complex, nutritious, and stress-tolerant food that has been used in Andean Indigenous cultures for thousands of years. Over the past several decades, numerous nutraceutical and food companies are using quinoa because of its perceived health benefits. Seeds of quinoa have a superb balance of proteins, lipids, carbohydrates, saponins, vitamins, phenolics, minerals, phytoecdysteroids, glycine betaine, and betalains. Quinoa due to its high nutritional protein contents, minerals, secondary metabolites and lack of gluten, is used as the main food source worldwide. In upcoming years, the frequency of extreme events and climatic variations is projected to increase which will have an impact on reliable and safe production of food. Quinoa due to its high nutritional quality and adaptability has been suggested as a good candidate to offer increased food security in a world with increased climatic variations. Quinoa possesses an exceptional ability to grow and adapt in varied and contrasting environments, including drought, saline soil, cold, heat UV-B radiation, and heavy metals. Adaptations in salinity and drought are the most commonly studied stresses in quinoa and their genetic diversity associated with two stresses has been extensively elucidated. Because of the traditional wide-ranging cultivation area of quinoa, different quinoa cultivars are available that are specifically adapted for specific stress and with broad genetic variability. This review will give a brief overview of the various physiological, morphological and metabolic adaptations in response to several abiotic stresses. creator: Qura Tul Ain creator: Kiran Siddique creator: Sami Bawazeer creator: Iftikhar Ali creator: Maham Mazhar creator: Rabia Rasool creator: Bismillah Mubeen creator: Farman Ullah creator: Ahsanullah Unar creator: Tassadaq Hussain Jafar uri: https://doi.org/10.7717/peerj.14832 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Ain et al. title: Dense GM-CSFRα-expressing immune infiltration is allied with longer survival of intrahepatic cholangiocarcinoma patients link: https://peerj.com/articles/14883 last-modified: 2023-03-02 description: BackgroundIntrahepatic cholangiocarcinoma (iCCA) is a cancer arising from intrahepatic bile duct epithelium. An iCCA incidence is increasing worldwide; however, the outcome of the disease is dismal. The linkage between chronic inflammation and iCCA progression is well established, but the roles of granulocyte-macrophage colony-stimulating factor (GM-CSF) remain unrevealed. Thus, a better understanding of GM-CSF functions in CCA may provide an alternative approach to CCA treatment.MethodsDifferential GM-CSF and GM-CSFRα mRNA expressions in CCA tissues were investigated by Gene Expression Profiling Interactive Analysis (GEPIA) based on The Cancer Genome Atlas (TCGA) database. The protein expressions and localizations of GM-CSF and its cognate receptor (GM-CSFRα) in iCCA patients’ tissues were demonstrated by the immunohistochemistry (IHC) techniques. The survival analyses were performed using Kaplan-Meier survival analysis with log-rank test and Cox proportional hazard regression model for multivariate analysis. The GM-CSF productions and GM-CSFRα expressions on CCA cells were assessed by ELISA and flow cytometry. The effects of GM-CSF on CCA cell proliferation and migration were evaluated after recombinant human GM-CSF treatment. The relationship between GM-CSF or GM-CSFRα level and related immune cell infiltration was analyzed using the Tumor Immune Estimation Resource (TIMER).ResultsGEPIA analysis indicated GM-CSF and GM-CSFRα expressions were higher in CCA tissues than in normal counterparts, and high GM-CSFRα was related to the longer disease-free survival of the patients (p < 0.001). IHC analysis revealed that CCA cells differentially expressed GM-CSF, while GM-CSFRα was expressed on cancer-infiltrating immune cells. The patient whose CCA tissue contained high GM-CSF expressed CCA, and moderate to dense GM-CSFRα-expressing immune cell infiltration (ICI) acquired longer overall survival (OS) (p = 0.047), whereas light GM-CSFRα-expressing ICI contributed to an increased hazard ratio (HR) to 1.882 (95% CI [1.077–3.287]; p = 0.026). In non-papillary subtype, an aggressive CCA subtype, patients with light GM-CSFRα-expressing ICI had shorter median OS (181 vs. 351 days; p = 0.002) and the HR was elevated to 2.788 (95% CI [1.299–5.985]; p = 0.009). Additionally, TIMER analysis demonstrated GM-CSFRα expression was positively correlated with neutrophil, dendritic cell, and CD8+ T cell infiltrations, though it was conversely related to M2-macrophage and myeloid-derived suppressor cell infiltration. However, the direct effects of GM-CSF on CCA cell proliferation and migration were not observed in the current study.ConclusionsLight GM-CSFRα-expressing ICI was an independent poor prognostic factor for iCCA patients. Anti-cancer functions of GM-CSFRα-expressing ICI were suggested. Altogether, the benefits of acquired GM-CSFRα-expressing ICI and GM-CSF for CCA treatment are proposed herein and require elucidation. creator: Paksiree Saranaruk creator: Sakda Waraasawapati creator: Yaovalux Chamgramol creator: Kanlayanee Sawanyawisuth creator: Natnicha Paungpan creator: Narumon Somphud creator: Chaisiri Wongkham creator: Seiji Okada creator: Sopit Wongkham creator: Kulthida Vaeteewoottacharn uri: https://doi.org/10.7717/peerj.14883 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Saranaruk et al. title: SARS-CoV-2 Delta (B.1.617.2) variant replicates and induces syncytia formation in human induced pluripotent stem cell-derived macrophages link: https://peerj.com/articles/14918 last-modified: 2023-03-02 description: Alveolar macrophages are tissue-resident immune cells that protect epithelial cells in the alveoli from invasion by pathogens, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Therefore, the interaction between macrophages and SARS-CoV-2 is inevitable. However, little is known about the role of macrophages in SARS-CoV-2 infection. Here, we generated macrophages from human induced pluripotent stem cells (hiPSCs) to investigate the susceptibility of hiPSC-derived macrophages (iMΦ) to the authentic SARS-CoV-2 Delta (B.1.617.2) and Omicron (B.1.1.529) variants as well as their gene expression profiles of proinflammatory cytokines during infection. With undetectable angiotensin-converting enzyme 2 (ACE2) mRNA and protein expression, iMΦ were susceptible to productive infection with the Delta variant, whereas infection of iMΦ with the Omicron variant was abortive. Interestingly, Delta induced cell-cell fusion or syncytia formation in iMΦ, which was not observed in Omicron-infected cells. However, iMΦ expressed moderate levels of proinflammatory cytokine genes in response to SARS-CoV-2 infection, in contrast to strong upregulation of these cytokine genes in response to polarization by lipopolysaccharide (LPS) and interferon-gamma (IFN-γ). Overall, our findings indicate that the SARS-CoV-2 Delta variant can replicate and cause syncytia formation in macrophages, suggesting that the Delta variant can enter cells with undetectable ACE2 levels and exhibit greater fusogenicity. creator: Theeradej Thaweerattanasinp creator: Asawin Wanitchang creator: Janya Saenboonrueng creator: Kanjana Srisutthisamphan creator: Nanchaya Wanasen creator: Suttipun Sungsuwan creator: Anan Jongkaewwattana creator: Thanathom Chailangkarn uri: https://doi.org/10.7717/peerj.14918 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Thaweerattanasinp et al. title: Hydroxy proline and gamma-aminobutyric acid: markers of susceptibility to vine decline disease caused by the fungus Monosporascus cannonballus in melons (Cucumis melo L.) link: https://peerj.com/articles/14932 last-modified: 2023-03-02 description: BackgroundVine decline disease caused by the fungus Monosporascus cannonballus, is a threat to melon production (Cucumis melo L.) worldwide. Nonetheless, little is known about the metabolites produced during the host pathogen interaction. Thus, the objective of this study was to measure quantities of amino acids produced over time during such an interaction.MethodsTwo melon genotypes named TAM-Uvalde (susceptible) and USDA PI 124104 (resistant) were grown and inoculated with M. cannonballus. The metabolites previously stated were measured before inoculation (0 hours) and 24, 48 and 72 hours after inoculation, using high performance liquid chromatography analysis.ResultsThe production of some amino acids during the interaction of the resistant and susceptible melon genotypes with the fungus M. cannonballus was different regarding quantities over time. Interestingly, hydroxy proline was always up-regulated in higher quantities in response to pathogen infection in the genotype TAM-Uvalde. Also, the up-regulation in higher quantities of gamma-aminobutyric acid in the genotype TAM-Uvalde 48 and 72 hours after inoculation, suggests more penetration of the pathogen in its roots. Hence, taken together, hydroxy proline and gamma-aminobutyric acid levels could be used as markers of susceptibility to vine decline disease caused by M. cannonballus, which could be useful in developing resistant varieties. creator: Sixto Alberto Marquez creator: Kevin Crosby creator: Bhimanagouda Patil creator: Carlos Avila creator: Amir MH Ibrahim creator: Herika Pessoa creator: Jashbir Singh uri: https://doi.org/10.7717/peerj.14932 license: https://creativecommons.org/publicdomain/zero/1.0/ rights: title: Biocontrol endophytes Bacillus subtilis R31 influence the quality, transcriptome and metabolome of sweet corn link: https://peerj.com/articles/14967 last-modified: 2023-03-02 description: During colonization of soil and plants, biocontrol bacteria can effectively regulate the physiological metabolism of plants and induce disease resistance. To illustrate the influence of Bacillus subtilis R31 on the quality, transcriptome and metabolome of sweet corn, field studies were conducted at a corn experimental base in Zhuhai City. The results show that, after application of B. subtilis R31, sweet corn was more fruitful, with a 18.3 cm ear length, 5.0 cm ear diameter, 0.4 bald head, 403.9 g fresh weight of single bud, 272.0 g net weight of single ear, and 16.5 kernels sweetness. Combined transcriptomic and metabolomic analyses indicate that differentially expressed genes related to plant-pathogen interactions, MAPK signaling pathway-plant, phenylpropanoid biosynthesis, and flavonoid biosynthesis were significantly enriched. Moreover, the 110 upregulated DAMs were mainly involved in the flavonoid biosynthesis and flavone and flavonol biosynthesis pathways. Our study provides a foundation for investigating the molecular mechanisms by which biocontrol bacteria enhance crop nutrition and taste through biological means or genetic engineering at the molecular level. creator: Mingwei Shao creator: Yanhong Chen creator: Qingyou Gong creator: Shuang Miao creator: Chunji Li creator: Yunhao Sun creator: Di Qin creator: Xiaojian Guo creator: Xun Yan creator: Ping Cheng creator: Guohui Yu uri: https://doi.org/10.7717/peerj.14967 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Shao et al. title: LincRNA00612 inhibits apoptosis and inflammation in LPS-induced BEAS-2B cells via enhancing interaction between p-STAT3 and A2M promoter link: https://peerj.com/articles/14986 last-modified: 2023-03-02 description: Long non-coding RNAs (lncRNAs) have been reported as key regulators of chronic obstructive pulmonary disease (COPD). This study aimed to figure out the regulatory mechanism as well as the effects of lncRNA00612 (LINC00612) in lipopolysaccharide (LPS)-induced inflammation and apoptosis in BEAS-2B cells. LINC00612 and its co-expressed gene alpha-2-macroglobulin (A2M) were strikingly downregulated in the peripheral venous blood of COPD patients. Overexpressed LINC00612 enhances BEAS-2B cells against apoptosis and inflammatory reactions mediated by LPS, however, an A2M knockdown can attenuate the degree of the enhancement. Bioinformatics analysis revealed putative binding sites between LINC00612, signal transducer and activator of transcription 3 (STAT3) and the A2M promoter, while RNA antisense purification and Chromatin immunoprecipitation were performed to confirm the prediction. Knockdown of LINC00612 impaired the binding of p-STAT3 to the promoter of A2M, which meant that LINC00612 was critical for the binding of STAT3 with the A2M promoter. Therefore, it can be concluded that LINC00612 ameliorates LPS-induced cell apoptosis and inflammation via recruiting STAT3 to bind to A2M. This conclusion will serve as a theoretical foundation for the treatment of COPD. creator: Xinru Xiao creator: Wei Cai creator: Ziqi Ding creator: Zhengdao Mao creator: Yujia Shi creator: Qian Zhang uri: https://doi.org/10.7717/peerj.14986 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Xiao et al. title: Plant ingredients in Thai food: a well-rounded diet for natural bioactive associated with medicinal properties link: https://peerj.com/articles/14568 last-modified: 2023-03-01 description: BackgroundSeeking cures for chronic inflammation-associated diseases and infectious diseases caused by critical human pathogens is challenging and time-consuming. Even as the research community searches for novel bioactive agents, consuming a healthy diet with functional ability might be an effective way to delay and prevent the progression of severe health conditions. Many plant ingredients in Thai food are considered medicinal, and these vegetables, herbs, and spices collectively possess multiple biological and pharmacological activities, such as anti-inflammatory, antimicrobial, antidiabetic, antipyretic, anticancer, hepatoprotective, and cardioprotective effects.MethodologyIn this review, the selected edible plants are unspecific to Thai food, but our unique blend of recipes and preparation techniques make traditional Thai food healthy and functional. We searched three electronic databases: PUBMED, Science Direct, and Google Scholar, using the specific keywords “Plant name” followed by “Anti-inflammatory” or “Antibacterial” or “Antiviral” and focusing on articles published between 2017 and 2021.ResultsOur selection of 69 edible and medicinal plant species (33 families) is the most comprehensive compilation of Thai food sources demonstrating biological activities to date. Focusing on articles published between 2017 and 2021, we identified a total of 245 scientific articles that have reported main compounds, traditional uses, and pharmacological and biological activities from plant parts of the selected species.ConclusionsEvidence indicates that the selected plants contain bioactive compounds responsible for anti-inflammatory, antibacterial, and antiviral properties, suggesting these plants as potential sources for bioactive agents and suitable for consumption for health benefits. creator: Raveevatoo Buathong creator: Sutsawat Duangsrisai uri: https://doi.org/10.7717/peerj.14568 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Buathong and Duangsrisai title: Unveiling the evolutionary relationships and the high cryptic diversity in Andean rainfrogs (Craugastoridae: Pristimantis myersi group) link: https://peerj.com/articles/14715 last-modified: 2023-03-01 description: BackgroundPristimantis is the most diverse genus of terrestrial frogs. Historically, it has been divided into several phenetic groups in order to facilitate species identification. However, in light of phylogenetic analysis, many of these groups have been shown to be non-monophyletic, denoting a high degree of morphological convergence and limited number of diagnostic traits. In this study, we focus on the Pristimantis myersi group, an assemblage of small rainfrogs distributed throughout the Andes of Ecuador and Colombia, whose external morphology is highly conserved, and its species diversity and evolutionary relationships largely unknown.MethodsWe inferred a new phylogenetic hypothesis for the frog genus Pristimantis, including all available sequences of the mtDNA 16S rRNA, as well as new DNA sequences from 175 specimens. Our sampling included 19 of the 24 species currently recognized as part of the Pristimantis myersi group.ResultsOur new evolutionary hypothesis recovered the P. myersi group as non-monophyletic and composed of 16 species. Therefore, we exclude P. albujai, P. bicantus, P. sambalan, and P. nelsongalloi in order to preserve the monophyly of the group. We discovered at least eight candidate species, most of them hidden under the names of P. leoni, P. hectus, P. festae, P. gladiator, and P. ocreatus.DiscussionOur results reveal the occurrence of a high level of cryptic diversity to the species level within the P. myersi group and highlight the need to redefine some of its species and reassess their conservation status. We suggest that the conservation status of six species within the group need to be re-evaluated because they exhibit smaller distributions than previously thought; these species are: P. festae, P. gladiator, P. hectus, P. leoni, P. ocreatus, and P. pyrrhomerus. Finally, given that the Pristimantis myersi group, as defined in this work, is monophyletic and morphologically diagnosable, and that Trachyphrynus is an available name for the clade containing P. myersi, we implement Trachyphrynus as a formal subgenus name for the Pristimantis myersi group. creator: Daniela Franco-Mena creator: Juan M. Guayasamin creator: Diego Andrade-Brito creator: Mario H. Yánez-Muñoz creator: Fernando J.M. Rojas-Runjaic uri: https://doi.org/10.7717/peerj.14715 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Franco-Mena et al. title: Coronavirus fear of contagion and compliance with social distancing measures: evidence for a sample of a Spanish university community link: https://peerj.com/articles/14771 last-modified: 2023-03-01 description: BackgroundSocial distancing measures have been one of the core pillars of the strategy against COVID-19 in all the countries. This study aims at understanding what motivates behaviours and compliance with social distancing measures among students and workers from a Spanish public university.MethodsWe carry out two logistics models considering two different dependent variables: not maintaining social relation with non-cohabiting people and not to leave home except for emergencies (n = 507, sample is formed by students and workers from the University of Cantabria in the North of Spain).ResultsBeing very concerned about getting ill suggests higher risk of not maintaining social relation with non-cohabiting people. Getting older increase the probability of not leaving home except for emergencies as happens with those who are very concerned about getting ill. Young people often living with vulnerable older relatives may affect students’ behaviour.ConclusionsOur findings suggest that compliance with social distancing measures depends on several factors related to age, the number or kind of cohabiting people and level of concern about getting ill. Policies should address all these factors through a multidisciplinary perspective. creator: David Cantarero Prieto creator: Paloma Lanza-León creator: Patricia Moreno creator: Javier Lera creator: Francisco Gonzalez creator: Irene González Rodríguez creator: Carla Blázquez-Fernández uri: https://doi.org/10.7717/peerj.14771 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Cantarero Prieto et al. title: Effects of habitat factors on the plant diversity on naturally-restored wind farm slopes link: https://peerj.com/articles/14912 last-modified: 2023-03-01 description: This study investigated naturally-restored plant communities on wind farm slopes and analyzed the effects of various habitat factors on the plant diversity. The findings provide a technical support for the ecological restoration of mountainous slopes.Twenty-one slopes on five wind farms were selected and the characteristics of the habitat, including slope position, slope aspect, slope gradient, altitude, years since restoration, and plant communities, were recorded. The species richness of the plant communities and the vegetation diversity indexes of these wind farms were measured and calculated, including the Shannon-Wiener diversity index (H′), Pielou’s species evenness index (J), and Margalef’s richness index (R). The key factors influencing plant diversity were identified using a stepwise regression analysis. A total of 36 families, 54 genera, and 57 species of plants were identified in this study with the Gramineae, Compositae, Rosaceae, Liliaceae, and Juglandaceae families the mostly predominant. Cynodon dactylon, Rubus lambertianus Ser., and Lindera glauca were the dominant species of herbs, shrubs, and trees, respectively. The highest number of species were found on lower slopes, slopes with semi-sunny aspects, slopes with gradients 30–50°, elevation below 500 m, and on slopes with at least five years since restoration. The plant diversity H′ and R tended to be higher on lower slopes than on upper slopes, and higher on slopes with semi-shady aspects than on slopes with semi-sunny aspects (P < 0.05). Vegetation diversity increased with the years since restoration. Slope position and slope aspect were identified as the primary influencing factors, and the H′ and R indexes were major indicators of changes in plant diversity on mountainous slopes. creator: Pengcheng Wang creator: Hai Yu creator: Henglin Xiao creator: Juan Wan creator: Qiang Ma creator: Gaoliang Tao creator: Qin Wang creator: Weiwei Jiang creator: Li Ma uri: https://doi.org/10.7717/peerj.14912 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Wang et al. title: Usability and acceptance of crowd-based early warning of harmful algal blooms link: https://peerj.com/articles/14923 last-modified: 2023-03-01 description: Crowdsensing has become an alternative solution to physical sensors and apparatuses. Utilizing citizen science communities is undoubtedly a much cheaper solution. However, similar to other participatory-based applications, the willingness of community members to be actively involved is paramount to the success of implementation. This research investigated factors that affect the continual use intention of a crowd-based early warning system (CBEWS) to mitigate harmful algal blooms (HABs). This study applied the partial least square-structural equation modeling (PLS-SEM) using an augmented technology acceptance model (TAM). In addition to the native TAM variables, such as perceived ease of use and usefulness as well as attitude, other factors, including awareness, social influence, and reward, were also studied. Furthermore, the usability factor was examined, specifically using the System Usability Scale (SUS) score as a determinant. Results showed that usability positively affected the perceived ease of use. Moreover, perceived usefulness and awareness influenced users’ attitudes toward using CBEWS. Meanwhile, the reward had no significant effects on continual use intention. creator: Lindung Parningotan Manik creator: Hatim Albasri creator: Reny Puspasari creator: Aris Yaman creator: Shidiq Al Hakim creator: Al Hafiz Akbar Maulana Siagian creator: Siti Kania Kushadiani creator: Slamet Riyanto creator: Foni Agus Setiawan creator: Lolita Thesiana creator: Meuthia Aula Jabbar creator: Ramadhona Saville creator: Masaaki Wada uri: https://doi.org/10.7717/peerj.14923 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2023 Manik et al. title: Incorporating dietary fiber from fruit and vegetable waste in meat products: a systematic approach for sustainable meat processing and improving the functional, nutritional and health attributes link: https://peerj.com/articles/14977 last-modified: 2023-03-01 description: BackgroundEvery year, the food business produces a sizeable amount of waste, including the portions of fruits and vegetables that are inedible, and those that have reached a stage where they are no longer suitable for human consumption. These by-products comprise of components such as natural antioxidants (polyphenols, carotenoid etc.), dietary fiber, and other trace elements, which can provide functionality to food. Due to changing lifestyles, there is an increased demand for ready-to-eat products like sausages, salami, and meat patties. In this line, meat products like buffalo meat sausages and patties are also gaining the interest of consumers because of their rich taste. Meat, however, has a high percentage of fat and is totally deprived of dietary fiber, which poses severe health problems like cardiovascular (CV) and gastrointestinal diseases. The health-conscious consumer is becoming increasingly aware of the importance of balancing flavor and nutrition. Therefore, to overcome this problem, several fruit and vegetable wastes from their respective industries can be successfully incorporated into meat products that provide dietary fiber and play the role of natural antioxidants; this will slow down lipid oxidation and increase the shelf-life of meat products.MethodologyExtensive literature searches have been performed using various scientific search engines. We collected relevant and informative data from subject-specific and recent literature on sustainable food processing of wasted food products. We also looked into the various applications of waste fruit and vegetable products, including cereals, when they are incorporated into meat and meat products. All relevant searches meeting the criteria were included in this review, and exclusion criteria were also set.ResultsThe pomace and peels of fruits like grapes, pomegranates, cauliflower, sweet lime, and other citrus are some of the most commonly used fruit and vegetable by-products. These vegetable by-products help inhibit oxidation (of both lipids and proteins) and the growth of pathogenic and spoilage bacteria, all without altering the consumer’s acceptability of the product on a sensory level. When included in meat products, these by-products have the potential to improve the overall product quality and lengthen its shelf-life under certain circumstances.ConclusionCost-effective and easily accessible by-products from the fruit and vegetable processing industries can be used in meat products to enhance their quality features (physicochemical, microbial, sensory, and textural aspects) and health benefits. Additionally, this will provides environmental food sustainability by lowering waste disposal and improving the food’s functional efficacy. creator: Abdul Haque creator: Saghir Ahmad creator: Z. R. A. A. Azad creator: Mohd Adnan creator: Syed Amir Ashraf uri: https://doi.org/10.7717/peerj.14977 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2023 Haque et al.