title: PeerJ description: Articles published in PeerJ link: https://peerj.com/articles/index.rss3?journal=peerj&page=617 creator: info@peerj.com PeerJ errorsTo: info@peerj.com PeerJ language: en title: M1 macrophage-derived extracellular vesicle containing tsRNA-5006c promotes osteogenic differentiation of aortic valve interstitial cells through regulating mitophagy link: https://peerj.com/articles/14307 last-modified: 2022-12-02 description: BackgroundOsteogenic differentiation of aortic valve interstitial cells (AVICs) plays a key role in the calcific aortic valve disease progression. Extracellular vesicles (EVs)-derived from M1-polarized macrophages (M1-EVs) orchestrated intercellular communication by delivering non-coding RNAs such as tRNA-derived small RNAs (tsRNAs) is crucial for cardiovascular disease. However, the role and mechanism of M1-EVs tsRNAs in osteogenic differentiation of AVICs remains largely unclear.MethodsM1-EVs and PBS treated-RAW 264.7 cell-derived EVs (NC-EVs) were incubated with AVICs and subjected to small RNA sequencing. Candidate tsRNA in M1-EVs was silenced to explore their effects on AVIC osteogenic differentiation and mitophagy.ResultsDiI-labeled M1-EVs were internalized by AVICs, resulting in significantly increased calcium nodule formation and expression of osteogenesis-related genes in AVICs, including RUNX2, BMP2, osteopontin, and SPP1, compared with NC-EVs. Small RNA sequencing revealed that 17 tsRNAs were significantly up-regulated such as tsRNA-5006c, while 28 tsRNAs were significantly down-regulated in M1-EVs compared with NC-EVs. Intriguingly, tsRNA-5006c-deleted M1-EVs treatment significantly reduced calcium nodule formation and expression of osteogenesis-related genes in AVICs relative to control group. Moreover, target genes of tsRNA-5006c were mainly involved in autophagy-related signaling pathways, such as MAPK, Ras, Wnt, and Hippo signaling pathway. Hallmarks of mitophagy activation in AVICs including mitophagosome formation, TMRM fluorescence, expression of LC3-II, BINP3, and PGC1α, were significantly elevated in the M1-EVs group compared with NC-EVs group, whereas M1-EVs tsRNA-5006c inhibitor led to a significant reduction in these indicators.ConclusionM1-EVs carried tsRNA-5006c regulates AVIC osteogenic differentiation from the perspective of mitophagy, and we provide a new target for the prevention and treatment of aortic valve calcification. creator: Hao Xia creator: Mingjian Gao creator: Jun Chen creator: Guanshen Huang creator: Xiuting Xiang creator: Yuyan Wang creator: Zhaohui Huang creator: Yongchun Li creator: Shuang Su creator: Zewei Zhao creator: Qingchun Zeng creator: Yunjun Ruan uri: https://doi.org/10.7717/peerj.14307 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2022 Xia et al. title: Effects of PAHs on meiofauna from three estuaries with different levels of urbanization in the South Atlantic link: https://peerj.com/articles/14407 last-modified: 2022-12-02 description: Estuarine environments are suggested to be the final receivers of human pollution and are impacted by surrounding urbanization and compounds carried by the river waters that flow from the continent. Polycyclic aromatic hydrocarbons (PAHs) are among the contaminants that can reach estuaries and can directly affect marine conservation, being considered highly deleterious to organisms living in these environments. This research investigated the meiofauna of three estuaries exposed to different levels of urbanization and consequently different levels of PAH concentrations, in order to assess how these compounds and environmental factors affect the distribution, structure and diversity of these interstitial invertebrates. A total of 15 major meiofauna groups were identified, with Nematoda being the dominant taxon (74.64%), followed by Copepoda (9.55%) and Polychaeta (8.56%). It was possible to observe significant differences in all diversity indices studied in the estuaries. With the exception of average density, the diversity indices (richness, Shannon index and evenness) were higher in the reference estuary, Goiana estuarine system (GES). On the other hand, the Timbó estuarine system (TES) had the lowest Shannon index value and richness, while the Capibaribe estuarine system (CES) had the lowest evenness value. The latter two estuaries (TES and CES) presented intermediate and high levels of urbanization, respectively. The ecological quality assessment (EcoQ) in the studied estuaries was classified from Poor to Moderate and the estuary with the lowest demographic density in its surroundings, GES, showed a better ecological quality (Moderate EcoQ). A significant distance-based multivariate linear modelling regression (DistLM) was observed between the environmental variables and the density of the meiobenthic community, where PAHs and pH were the main contributors to organism variation. The sediments were characterized by predominance of very fine sand and silt-clay in the most polluted environments, while the control site environment (GES) was dominated by medium grains. The highest concentrations of PAHs were found in the most urbanized estuaries, and directly affected the structure of the interstitial benthic community. The metrics used in the present study proved to be adequate for assessing the environmental quality of the investigated estuaries. creator: Renan B. da Silva creator: Giovanni A. P. Dos Santos creator: Ana Luiza L. de Farias creator: Débora A. A. França creator: Raianne Amorim Cavalcante creator: Eliete Zanardi-Lamardo creator: Jose Roberto Botelho de Souza creator: Andre M. Esteves uri: https://doi.org/10.7717/peerj.14407 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2022 da Silva et al. title: Transcriptome-wide identification of WRKY transcription factors and their expression profiles under different stress in Cynanchum thesioides link: https://peerj.com/articles/14436 last-modified: 2022-12-02 description: Cynanchum thesioides (Freyn) K. Schum. is an important economic and medicinal plant widely distributed in northern China. WRKY transcription factors (TFs) play important roles in plant growth, development and regulating responses. However, there is no report on the WRKY genes in Cynanchum thesioides. A total of 19 WRKY transcriptome sequences with complete ORFs were identified as WRKY transcriptome sequences by searching for WRKYs in RNA sequencing data. Then, the WRKY genes were classified by phylogenetic and conserved motif analysis of the WRKY family in Cynanchum thesioides and Arabidopsis thaliana. qRT–PCR was used to determine the expression patterns of 19 CtWRKY genes in different tissues and seedlings of Cynanchum thesioides under plant hormone (ABA and ETH) and abiotic stresses (cold and salt). The results showed that 19 CtWRKY genes could be divided into groups I-III according to their structure and phylogenetic characteristics, and group II could be divided into five subgroups. The prediction of CtWRKY gene protein interactions indicates that CtWRKY is involved in many biological processes. In addition, the CtWRKY gene was differentially expressed in different tissues and positively responded to abiotic stress and phytohormone treatment, among which CtWRKY9, CtWRKY18, and CtWRKY19 were significantly induced under various stresses. This study is the first to identify the WRKY gene family in Cynanchum thesioides, and the systematic analysis lays a foundation for further identification of the function of WRKY genes in Cynanchum thesioides. creator: Xiaoyao Chang creator: Zhongren Yang creator: Xiaoyan Zhang creator: Fenglan Zhang creator: Xiumei Huang creator: Xu Han uri: https://doi.org/10.7717/peerj.14436 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2022 Chang et al. title: Multi-omics analyses reveal that the gut microbiome and its metabolites promote milk fat synthesis in Zhongdian yak cows link: https://peerj.com/articles/14444 last-modified: 2022-12-02 description: BackgroundYak cows produce higher quality milk with higher concentrations of milk fat than dairy cows. Recently, studies have found the yak milk yield and milk fat percentage have decreased significantly over the past decade, highlighting the urgency for yak milk improvement. Therefore, we aimed to analyze how the gut microbiome impacts milk fat synthesis in Zhongdian yak cows.MethodsWe collected milk samples from Zhongdian yak cows and analyzed the milk fat percentage, selecting five Zhongdian yak cows with a very high milk fat percentage (>7%, 8.70 ± 1.89%, H group) and five Zhongdian yak cows with a very low milk fat percentage (<5%, 4.12 ± 0.43%, L group), and then obtained gut samples of these ten Zhongdian yak cows through rectal palpation. Gut metagenomics, metabolomics, and conjoint metagenomics and metabolomics analyses were performed on these samples, identifying taxonomic changes, functional changes, and changes in gut microbes-metabolite interactions within the milk fat synthesis-associated Zhongdian yak cows gut microbiome, to identify potential regulatory mechanisms of milk fat at the gut microbiome level in Zhongdian yak cows.ResultsThe metagenomics analysis revealed Firmicutes and Proteobacteria were significantly more abundant in the gut of the high-milk fat Zhongdian yak cows. These bacteria are involved in the biosynthesis of unsaturated fatty acids and amino acids, leading to greater efficiency in converting energy to milk fat. The metabolomics analysis showed that the elevated gut metabolites in high milk fat percentage Zhongdian yak cows were mainly enriched in lipid and amino acid metabolism. Using a combined metagenomic and metabolomics analysis, positive correlations between Firmicutes (Desulfocucumis, Anaerotignum, Dolosiccus) and myristic acid, and Proteobacteria (Catenovulum, Comamonas, Rubrivivax, Marivita, Succinimouas) and choline were found in the gut of Zhongdian yak cows. These interactions may be the main contributors to methanogen inhibition, producing less methane leading to higher-efficient milk fat production.ConclusionsA study of the gut microbe, gut metabolites, and milk fat percentage of Zhongdian yak cows revealed that the variations in milk fat percentage between yak cows may be caused by the gut microbes and their metabolites, especially Firmicutes-myristic acid and Proteobacteria-choline interactions, which are important to milk fat synthesis. Our study provides new insights into the functional roles of the gut microbiome in producing small molecule metabolites and contributing to milk performance traits in yak cows. creator: Lily Liu creator: Peifu Wu creator: Fenfen Chen creator: Jielong Zhou creator: Aiwei Guo creator: Kerong Shi creator: Qin Zhang uri: https://doi.org/10.7717/peerj.14444 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2022 Liu et al. title: Genome-wide identification and characterization of NHL gene family in response to alkaline stress, ABA and MEJA treatments in wild soybean (Glycine soja) link: https://peerj.com/articles/14451 last-modified: 2022-12-02 description: BackgroundNDR1/HIN1-like (NHL) family genes are known to be involved in pathogen induced plant responses to biotic stress. Even though the NHL family genes have been identified and characterized in plant defense responses in some plants, the roles of these genes associated with the plant abiotic stress tolerance in wild soybean is not fully established yet, especially in response to alkaline stress.MethodsWe identified the potential NHL family genes by using the Hidden Markov model and wild soybean genome. The maximum-likelihood phylogenetic tree and conserved motifs were generated by using the MEME online server and MEGA 7.0 software, respectively. Furthermore, the syntenic analysis was generated with Circos-0.69. Then we used the PlantCARE online software to predict and analyze the regulatory cis-acting elements in promoter regions. Hierarchical clustering trees was generated using TM4: MeV4.9 software. Additionally, the expression levels of NHL family genes under alkaline stress, ABA and MEJA treatment were identified by qRT-PCR.ResultsIn this study, we identified 59 potential NHL family genes in wild soybean. We identified that wild soybean NHL family genes could be mainly classified into five groups as well as exist with conserved motifs. Syntenic analysis of NHL family genes revealed genes location on 18 chromosomes and presence of 65 pairs of duplication genes. Moreover, NHL family genes consisted of a variety of putative hormone-related and abiotic stress responsive elements, where numbers of methyl jasmonate (MeJA) and abscisic acid (ABA) responsive elements were significantly larger than other elements. We confirmed the regulatory roles of NHL family genes in response to alkaline stress, ABA and MEJA treatment. In conclusion, we identified and provided valuable information on the wild soybean NHL family genes, and established a foundation to further explore the potential roles of NHL family genes in crosstalk with MeJA or ABA signal transduction mechanisms under alkaline stress. creator: Xu Zhang creator: Yongguo Xue creator: Haihang Wang creator: Zaib_un Nisa creator: Xiaoxia Jin creator: Lijie Yu creator: Xinlei Liu creator: Yang Yu creator: Chao Chen uri: https://doi.org/10.7717/peerj.14451 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2022 Zhang et al. title: The effects of a set amount of regular maternal exercise during pregnancy on gut microbiota are diet-dependent in mice and do not cause significant diversity changes link: https://peerj.com/articles/14459 last-modified: 2022-12-02 description: BackgroundDiet and exercise can affect the gut microbiota (GM); however, the effects of the same amount of exercise on gut microbiota changes in people on a low-fat diet (LFD) and high-fat diet (HFD) during pregnancy are unknown. Do different nutritional conditions respond equally to exercise intervention? This study aimed to investigate the effects of regular maternal exercise during pregnancy on the GM in mice fed different diets during pregnancy.MethodsSix-week-old nulliparous female KunMing mice were fed either a HFD or LFD before and during pregnancy. Each group of mice were then randomly divided into two groups upon confirmation of pregnancy: sedentary (HFD or LFD; n = 4 and 5, respectively) and exercised (HFDex or LFDex, n = 5 and 6, respectively). Mice were sacrificed on day 19 of gestation and their colon contents were collected. We then performed 16S rDNA gene sequencing of the V3 and V4 regions of the GM.ResultsThe pregnancy success rate was 60% for LFDex and 100% for HFDex. Both Chao1 and Simpson indices were not significantly different for either LFD vs. LFDex or HFD vs. HFDex. Desulfobacterota, Desulfovibrionia Desulfovibrionales, Desulfovibrionaceae, Desulfovibrio, Coriobacteriia, Coriobacteriales, and Eggerthellaceae were markedly decreased after exercise intervention in LFDex vs. LFD, whereas Actinobacteria, Bifidobacteriales, Bifidobacteriaceae, Bifidobacterium, and Bifidobacterium pseudolongum were significantly increased in LFDex vs. LFD. Furthermore, decreased Peptostreptococcales-Tissierellales and Peptostreptococcaceae and increased Bacteroides dorei were identified in the HFDex vs. HFD group. p_Desulfobacterota, c_Desulfovibrionia, o_Desulfovibrionales, f_Desulfovibrionaceae and g_Desulfovibrio were markedly decreased in the LFDex group vs. HFDex group.ConclusionsOur data suggested that quantitative maternal exercise during pregnancy resulted in alterations in GM composition, but did not significantly change the diversity of the GM. These findings may have important implications when considering an individual’s overall diet when recommending exercise during pregnancy. creator: Xia Duan creator: Jingjing Xu creator: Ping Yang creator: Xinyuan Liang creator: Zichun Zeng creator: Huijuan Luo creator: Xiaomei Tang creator: Xin Wu creator: Xiaomin Xiao uri: https://doi.org/10.7717/peerj.14459 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2022 Duan et al. title: Cathepsin B plays a role in spermatogenesis and sperm maturation through regulating autophagy and apoptosis in mice link: https://peerj.com/articles/14472 last-modified: 2022-12-02 description: Spermatogenesis and sperm maturation are complex and highly ordered biological processes. Any failure or disorder in these processes can cause defects in sperm morphology, motility, and fertilization ability. Cathepsin B (CTSB) is involved in the regulation of a variety of pathological processes. In the present study, we found that CTSB was abundantly expressed in the male reproductive system, however, the specific role of CTSB in regulating spermatogenesis and sperm maturation remained elusive. Hence, we generated Ctsb-/- mice using CRISPR/Cas9 technology. In Ctsb-/- mice, sperm count was significantly decreased while the level of morphologically abnormal sperm was markedly increased. Additionally, these mice had significantly lower levels of progressive motility sperm and elevated levels of immobilized sperm. Histological analysis showed slight vacuolization in the testis epithelium, as well as the loss of epididymal epithelium cells. Further investigation showed that autophagic activity was inhibited and apoptotic activity was increased in both the testis and epididymis of Ctsb-/- mice. Together, our findings demonstrate that CTSB plays an important role in spermatogenesis and sperm maturation in mice. creator: Zongzhuang Wen creator: Haixia Zhu creator: Bin Wu creator: Aizhen Zhang creator: Hongxiang Wang creator: Yin Cheng creator: Hui Zhao creator: Jianyuan Li creator: Min Liu creator: Jiangang Gao uri: https://doi.org/10.7717/peerj.14472 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2022 Wen et al. title: Litter inputs and standing stocks in riparian zones and streams under secondary forest and managed and abandoned cocoa agroforestry systems link: https://peerj.com/articles/13787 last-modified: 2022-12-01 description: BackgroundCocoa is an important tropical tree crop that is mainly cultivated in agroforestry systems (AFS). This system, known as cabruca in northeastern Brazil, holds promise to reconcile biodiversity conservation and economic development. However, since cocoa AFS alters forest structure composition, it can affect litter dynamics in riparian zones and streams. Thus, our objective was to determine litter inputs and standing stocks in riparian zones and streams under three types of forest: managed cocoa AFS, abandoned cocoa AFS, and secondary forest.MethodsWe determined terrestrial litter fall (TI), vertical (VI) and lateral (LI) litter inputs to streams, and litter standing stocks on streambeds (BS) in the Atlantic Forest of northeastern Brazil. Litter was collected every 30 days from August 2018 to July 2019 using custom-made traps. The litter was dried, separated into four fractions (leaves, branches, reproductive organs, and miscellaneous material) and weighed.ResultsTerrestrial litter fall was similar in all forests, ranging from 89 g m−2 month−1 in secondary forest (SF) to 96 g m−2 month−1 in abandoned cocoa AFS (AC). Vertical input were higher in AC (82 g m−2 month−1) and MC (69 g m−2 month−1) than in SF (40 g m−2 month−1), whereas lateral input were higher in MC (43 g m−2 month−1) than in AC (15 g m−2 month−1) and SF (24 g m−2 month−1). Standing stocks followed the order SF > AC > MC, corresponding to 425, 299 and 152 g m−2. Leaves contributed most to all litter fractions in all forests. Reproductive plant parts accounted for a larger proportion in managed AFS. Branches and miscellaneous litter were also similar in all forests, except for higher benthic standing stocks of miscellaneous litter in the SF. Despite differences in the amounts of litter inputs and standing stocks among the forests, seasonal patterns in the abandoned AFS (AC) were more similar to those of the secondary forest (SF) than the managed AFS, suggesting potential of abandoned AFS to restore litter dynamics resembling those of secondary forests. creator: Haialla Carolina Rialli Santos Brandão creator: Camila Andrade Coqueiro Moraes creator: Ana Paula Silva creator: José Francisco Gonçalves Júnior creator: Renan de Souza Rezende creator: Daniela Mariano Lopes da Silva uri: https://doi.org/10.7717/peerj.13787 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2022 Rialli Santos Brandão et al. title: Establishment of the non-native horned-face bee Osmia cornifrons and the taurus mason bee Osmia taurus (Hymenoptera: Megachilidae) in Canada link: https://peerj.com/articles/14216 last-modified: 2022-12-01 description: Established populations of the non-native horned-face bee, Osmia cornifrons (Radoszkowski, 1887), and the taurus mason bee, Osmia taurus Smith, 1873 (Hymenoptera: Megachilidae), have been identified from Canada for the first time. In the US, the importation of O. cornifrons, beginning in the 1970s, led to its release for agricultural crop pollination and spread across the country. In this article, we report on O. cornifrons captured while sampling wild bees in Toronto, Ontario using hand nets, bug vacuums, and vane traps, as well as established populations in trap nests, from 2017–2020. The morphologically similar O. taurus, which was accidentally introduced to the US with shipments of imported O. cornifrons, was also recorded in our samples. Recently, a few individual O. taurus specimens have been identified from Ontario and Quebec; however, the extent of our sampling included nests, indicating it is also established in Canada. Others have shown its population growth to have been associated with concordant declines in abundances of native mason bee species in the US, and similar impacts are possible in Canada if action is not taken. We propose three non-mutually exclusive possible pathways for the arrival of O. cornifrons, as well as O. taurus, in Canada: (1) natural migration northward from non-native populations in the US, (2) international importation in the 1980s–2000s to support agricultural research programs, and (3) unintentional release of mason bee cocoons purchased from non-local vendors. We argue that a focus on enhancing populations of locally occurring native bees and stronger policy on the importation and sale of non-native bees are needed. creator: J. Scott MacIvor creator: Charlotte W. de Keyzer creator: Madison S. Marshall creator: Graham S. Thurston creator: Thomas M. Onuferko uri: https://doi.org/10.7717/peerj.14216 license: https://creativecommons.org/licenses/by/4.0/ rights: © 2022 MacIvor et al. title: Level of anxiety and depression among healthcare workers in Saudi Arabia during the COVID-19 pandemic link: https://peerj.com/articles/14246 last-modified: 2022-12-01 description: BackgroundThe coronavirus disease 2019 (COVID-19) pandemic places a high demand on frontline healthcare workers. Healthcare workers are at high-risk of contracting the virus and are subjected to its consequential emotional and psychological effects. This study aimed to measure the level of depression and anxiety among healthcare workers in Saudi Arabia during the early stages of the COVID-19 pandemic.MethodsThis was a cross-sectional study; data were collected from healthcare workers in Saudi Arabia using a survey that included the Zung Self-Rating Depression Scale and the Generalized Anxiety Disorder Scale-7. A total of 326 participants took part in the study by completing and submitting the survey.ResultsThe vast majority of the participating healthcare workers were Saudi nationals (98.8%) working in a public healthcare facility (89.9%). The results indicated that most of the participants had mild levels of anxiety and depression. A total of 72.5% of the respondents had anxiety, ranging from mild (44.1%) to moderate (16.2%) and severe (12.2%). Moreover, 24.4% of the respondents had depression ranging from mild (21.7%) to moderate (2.1%) and severe (0.6%). The generalized linear models showed that the <30 age group (Beta = 0.556, p = 0.037) and the 30–39-year age group (Beta = 0.623, p = 0.019) were predicted to have anxiety. The analysis revealed that females were more anxious (Beta = 0.241, p = 0.005) than males. Healthcare providers working in primary healthcare centers (Beta = −0.315, p = 0.008) and labs (Beta = −0.845. p = 0.0001 were predicted to be less anxious than those working in other healthcare facilities. The data analysis showed that participants with good economic status had more depression than the participants in the other economic status groups (Beta = 0.067, p = 0.003).ConclusionThis study found that the level of anxiety and depression in healthcare workers was mild. The factors that may contribute to anxiety in healthcare workers included being female, being younger than 30 or between the ages of 31 and 39, working in a specialized hospital facility, and the number of COVID-19 cases the workers dealt with. Economic status was associated with depression. A longitudinal study design is needed to understand the pattern of anxiety levels among healthcare workers over time during the COVID-19 pandemic. creator: Abbas Shamsan creator: Mohammed Alhajji creator: Yasmine Alabbasi creator: Ali Rabaan creator: Saad Alhumaid creator: Mansour Awad creator: Abbas Al Mutair uri: https://doi.org/10.7717/peerj.14246 license: https://creativecommons.org/licenses/by/4.0/ rights: ©2022 Shamsan et al.