The effects of naproxen sodium on the growth, reproduction, survival, and feeding of a freshwater pond snail
Over-the-counter drugs are emerging contaminants in the environment. These contaminants often affect aquatic communities, including freshwater invertebrates. Nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly found in aquatic ecosystems worldwide and have documented negative effects on aquatic organisms. Naproxen is often suggested as a more environmentally friendly alternative due to less bioconcentration in fish. However, studies have yet to test the effects of naproxen on freshwater molluscs. This study exposed pond snails, Physa spp., to three nominal levels of naproxen sodium, 100 µg/L, 1,000 µg/L and 10,000 µg/L, along with a control, over a month-long experiment. We measured snail growth, survival, reproduction, and feeding. Naproxen reduced snail survival across all levels of naproxen exposure and the rate that snails fed at the highest level of naproxen. However, we did not detect an effect of naproxen on snail growth or reproduction rates, even at pollution levels well above those observed in natural systems. Naproxen reduced the grazing of Physa spp. only at our highest concentration, but reduced survival by at least 50% across all concentrations.
Environmental Contamination and Remediation
Becker et al. (2025) The effects of naproxen sodium on the growth, reproduction, survival, and feeding of a freshwater pond snail. PeerJ 13:e20163 https://doi.org/10.7717/peerj.20163
Prediction of ACL injury incidence and analysis of key features in basketball players based on multi-algorithm models
Background Basketball players are a high-risk group for anterior cruciate ligament (ACL) injuries. This study aimed to identify the critical factors contributing to ACL injuries in male basketball players and evaluate the performance of machine learning (ML) algorithms in injury prediction. Methodology This study protocol was registered with International Standard Registered Clinical/soCial sTudy Number (ISRCTN) (Registration number: 18009799). A total of 104 male collegiate basketball players volunteered to participate in this study. Data on the athletes’ profile, physical functions, basketball-specific skills, biomechanics, and electromyography (EMG) of seven lower limb muscles during unanticipated side-cutting maneuvers were collected. A 12-month follow-up was conducted to compare these variables between the injured (n = 11) and non-injured (n = 93) groups. Only the variables with significant differences between the groups were included in the predictive modeling. Results The performance of machine learning models in predicting ACL injury risk was assessed using the area under the curve (AUC) of the receiver operating characteristic (ROC). The AUC-ROC values ranged from 0.66 to 0.80, with the random forest algorithm achieving the highest performance (AUC-ROC = 0.80). The most influential predicting feature observed during the emergency stop phase, included a greater knee flexion moment, reduced knee flexion angle, increased backward ground reaction force, and increased activation of the vastus lateralis muscle. Conclusion The random forest model demonstrated superior predictive performance, providing valuable insights into the key risk factors associated with ACL injury among male basketball players. This study highlighted the importance of biomechanical testing based on sport-specific movements to accurately predict the ACL injury risk.
Guo et al. (2025) Prediction of ACL injury incidence and analysis of key features in basketball players based on multi-algorithm models. PeerJ 13:e20141 https://doi.org/10.7717/peerj.20141
Metabarcoding analysis reveals hidden eukaryotic plankton biodiversity in the Ross Sea, Antarctica
Background Environmental DNA (eDNA) analysis is a highly sensitive, non-destructive method that enables the detection of various species through DNA shed into environmental samples without requiring direct organism collection. This study sought to investigate the biodiversity and community structure of eukaryotic plankton, including phytoplankton and zooplankton, in the Ross Sea marine protected area (RSR MPA) using eDNA metabarcoding analysis. By examining their spatial and vertical distributions, the study underscores the importance of continuous monitoring for the conservation of the RSR MPA. Methods We collected 48 seawater samples from 16 sites in the Ross Sea region for eDNA metabarcoding analysis, targeting the 18S rRNA gene region of eukaryotic plankton in Antarctica. Bioinformatic processing and taxonomic classification were conducted to assess the diversity and community composition of phytoplankton and zooplankton. Results Phytoplankton communities were primarily composed of six phyla with their distribution patterns and the grouping of samples with similar community structures was found to be shaped by the ocean currents of the RSR MPA and various environmental factors, such as salinity and dissolved oxygen levels. Zooplankton communities consisted of 18 major taxonomic groups, exhibiting distinct horizontal and vertical distribution patterns with differences in taxonomic community structure and species diversity across depth groups. Notably, previously undetected Antarctic species were identified in the Ross Sea region, demonstrating the effectiveness of eDNA in revealing hidden biodiversity. Conclusions Analyzing eukaryotic plankton communities in the vast and extreme Antarctic environment based on eDNA has proven to be highly efficient, enabling the detection of a greater number of species, including those that were difficult to identify in previous studies. It was observed that in the Ross Sea Marine Protected Area, various species form distinct community structures such as phytoplankton and zooplankton, each inhabiting the area according to different environmental variables and habitat preferences. As a designated marine protected area, the Ross Sea’s unique ecosystem requires continuous monitoring and conservation efforts to address environmental changes. The genetic data obtained in this study contributes to expanding the database of Antarctic-specific species, facilitating more accurate and efficient analyses of Antarctic ecosystems in the future.
Choi et al. (2025) Metabarcoding analysis reveals hidden eukaryotic plankton biodiversity in the Ross Sea, Antarctica. PeerJ 13:e20118 https://doi.org/10.7717/peerj.20118
Identification of diabetes related phenotype and diagnostic biomarkers in coronary artery disease
Background Diabetes mellitus (DM) and coronary artery disease (CAD) are closely interrelated clinical conditions. However, the combination analysis based on DM related CAD diagnostic model remains a gap. The primary objective of this study was to identify diagnostic models and diagnostic markers for CAD based on the association of diabetic phenotypes and attempt to explore them further in a mouse model. Methods We used data integration as well as multiple datasets for both coronary artery disease and diabetes to exclude bias as well as to improve reliability. We employed the least absolute shrinkage and selection operator (LASSO) regression algorithms to construct the CAD diagnostic model. Furthermore, we established mouse CAD model (low-density lipoprotein receptor deficient mice with high fat diet) to explore the crosstalk between the screened biomarkers and severe CAD progress. Results The intersecting genes from differential analysis and weighted correlation network analysis (WGCNA) results yielded 32 diabetes-related biomarkers. We then identified two diabetes-related phenotypes through the consensus clustering in CAD patients. Microenvironmental analysis revealed that phenotype 1 exhibited higher expression of most cytokines, inflammatory factors, interleukins, and related receptors. Immune cell composition in phenotype 1 showed increased infiltration compared to phenotype 2. The LASSO regression identified 16 diabetes-related genes and we further constructed a diagnostic model based on these genes, which the area under the curve (AUC) reached 0.8. Additionally, single cell immune analysis exhibited the location of these genes. KCNQ1, ATP6V1B1, MTDH, and ITPK1 were predominantly located in macrophages, indicating their potential in regulating macrophage during myocardial injury. Furthermore, We elucidated that KCNQ1 and ITPK1 exhibited high expression level in mouse CAD model in tissue level. exhibited similar expression trends with macrophage biomarkers (CD31 and CD68). The result of qPCR also indicated the elevated level of KCNQ1 and ITPK1, which exhibited crosstalk with CD31 and CD68 in mouse CAD model. Conclusion This study delves into the microenvironmental characteristics of diabetes-related phenotypes in CAD, constructing an optimal diagnostic model and validated the significance of diagnostic markers in mouse CAD model, which may offer insights that could be beneficial for clinical management in the near future.
Aihemaiti et al. (2025) Identification of diabetes related phenotype and diagnostic biomarkers in coronary artery disease. PeerJ 13:e20117 https://doi.org/10.7717/peerj.20117
Comparison of supervised versus home-based exercise programs following platelet-rich plasma injections in knee osteoarthritis: clinical outcomes from quasi-experimental study
Background Knee osteoarthritis (OA) is characterized by severe pain and decreased range of motion (ROM), mostly managed with conservative treatments. Therapies like platelet-rich plasma (PRP) intra-articular injections have gained fame for pain relief when surgery is a last resort. Objectives To compare the effectiveness of a supervised exercise program versus a home-based exercise program following PRP injections in terms of pain relief, ROM improvement, and disability reduction in knee osteoarthritis patients. Methods A quasi-experimental design was employed, enrolling 32 patients diagnosed with knee OA cases (age range: 40–60 years; 59% female), divided into two groups: a supervised exercise group (n = 16) and an unsupervised/home-based exercise group (n = 16). Both groups received two PRP injections spaced 15 days apart. The supervised group followed a structured exercise protocol, including cold packs post-injection for pain management. The program comprises three sessions per week for one month. The unsupervised group received similar exercises to be performed independently at home. Data analysis was conducted using parametric and non-parametric statistical methods based on data distribution. Results Both groups exhibited significant improvements from baseline to post-intervention in pain, ROM, and functional status (p < 0.05). The supervised exercise group demonstrated superior outcomes in pain reduction, knee flexion ROM, and Knee Injury and Osteoarthritis Outcome Score (KOOS). However, no significant difference was observed between groups for knee extension ROM (p = 0.378). Conclusion Both supervised and unsupervised exercise programs improved outcomes in knee OA patients following PRP injections; the supervised program showed greater pain relief, flexion ROM, and functional status. However, no significant difference was observed in knee extension ROM between the two groups.
Safdar et al. (2025) Comparison of supervised versus home-based exercise programs following platelet-rich plasma injections in knee osteoarthritis: clinical outcomes from quasi-experimental study. PeerJ 13:e19971 https://doi.org/10.7717/peerj.19971
Metabolic profile alterations in juvenile rats with bladder overactivity induced by short-term high-fructose intake
Background Overactive bladder syndrome (OAB) is a common condition that affects both adults and children, often causing significant discomfort and negatively impacting quality of life. This study aimed to preliminarily evaluate whether fructose exposure induces OAB-like symptoms in juvenile rats and to explore potential metabolic mechanisms involved. Methods Three-week-old female Sprague-Dawley rats were randomly assigned to two groups: a model group fed a 60% fructose diet for one week, and a control group fed a standard diet. Body weight, blood biochemical parameters, urination behavior, and urodynamic function were assessed. Bladder tissue was analyzed using hematoxylin and eosin staining, immunohistochemistry, and immunofluorescence. Molecular and metabolic changes were evaluated via Western blot and metabolomic analysis. Results Short-term high fructose intake led to significant metabolic and functional changes in juvenile rats. The model group showed reduced body weight and altered lipid metabolism, while glucose and insulin levels remained largely unchanged. Urodynamic assessments revealed increased non-voiding contractions, shortened intervals between contractions, and impaired bladder storage function. Open field tests indicated increased urinary frequency and open field test results. Histological analysis demonstrated disorganized bladder smooth muscle structure and elevated oxidative stress. Metabolomic profiling revealed marked alterations in energy metabolism, including enhanced glycolysis, pentose phosphate pathway activation, and accumulation of ATP and lactate. Western blot analysis confirmed activation of the CaMKK2/AMPK signaling pathway in bladder tissue. Conclusions Short-term high-fructose intake induces OAB-like symptoms in juvenile rats, accompanied by fructose toxicity, oxidative stress, calcium ion accumulation, and activation of the CaMKK2/AMPK signaling pathway. These findings highlight a potential link between fructose metabolism and bladder dysfunction, offering new insights for exploring metabolic or antioxidant-based therapeutic strategies in pediatric OAB.
Cui et al. (2025) Metabolic profile alterations in juvenile rats with bladder overactivity induced by short-term high-fructose intake. PeerJ 13:e20186 https://doi.org/10.7717/peerj.20186
rtpcr: a package for statistical analysis and graphical presentation of qPCR data in R
Background Quantitative real-time polymerase chain reaction (qRT-PCR or qPCR) is widely used in molecular biology research. Various analysis methods are employed to interpret qPCR data and measure mRNA levels of target genes under different experimental conditions. Results The rtpcr package was developed for amplification efficiency calculation, statistical analysis, and graphical presentation of qPCR data in R. It uses a general calculation methodology that accommodates up to two reference genes and amplification efficiency values, including the Pfaffl method. Depending on the experimental design, rtpcr functions apply a t-test (for experiments with a two-level factor), analysis of variance (ANOVA), or analysis of covariance (ANCOVA) (for experiments with more than two levels or factors) to calculate fold change (FC) or relative expression (RE) of a target gene. The functions also provide standard errors and confidence intervals for the means and support statistical mean comparisons. To facilitate usage, the package includes example datasets. It also offers ggplot-based visualizations with customizable arguments, allowing users to tailor the graphical output. Conclusions In summary, the rtpcr package is a useful and user-friendly tool for analyzing real-time PCR data from experiments involving up to three different factors. Built on a general methodology, it provides robust calculations and comprehensive graphical outputs, making it a valuable resource for researchers working with qPCR data.
Mirzaghaderi (2025) rtpcr: a package for statistical analysis and graphical presentation of qPCR data in R. PeerJ 13:e20185 https://doi.org/10.7717/peerj.20185
Extraction, identification and component analysis of exosome-like nanovesicles in Anoectochilus roxburghii (Wall.) Lindl.
Objective This study aimed to investigate the presence of exosome-like nanovesicles in Anoectochilus roxburghii (Wall.) Lindl. (A. roxburghii) and the related active ingredients. Methods A. roxburghii-derived exosome-like nanovesicles (ARENVs) were extracted by the differential ultracentrifugation and characterized by transmission electron microscopy and nano-flow cytometry. The components in ARENVs derived from soil-cultured and hydroponic A. roxburghii were identified and quantified by liquid chromatography-mass spectrometry. Results Nanovesicles with typical exosome characteristics were successfully isolated from A. roxburghii. After analysis and identification, ARENVs were rich in multiple flavonoid components such as isoquercitrin, kaempferol, isorhamnetin, narcissoside, quercetin and rutin. The quantity of flavonoid compounds was higher in ARENVs derived from soil-cultured A. roxburghii than in those from hydroponic A. roxburghii. Conclusion A. roxburghii contains exosome-like nanovesicles, which are rich in active flavonoid ingredients. These active ingredients are more abundant in ARENVs derived from soil-cultured A. roxburghii than hydroponic counterparts. The findings may provide an innovative theoretical basis for further exploration of the medicinal value of A. roxburghii.
Zhou et al. (2025) Extraction, identification and component analysis of exosome-like nanovesicles in Anoectochilus roxburghii (Wall.) Lindl.. PeerJ 13:e20182 https://doi.org/10.7717/peerj.20182
Association of sociodemographic factors, lifestyle behaviors, anthropometric measures, and cardiometabolic health markers with blood pressure in adolescents: a cross-sectional analysis
Introduction Evidence suggests that hypertension in adulthood may have its origins in childhood and adolescence. In this context, previous studies have demonstrated an association between lifestyle behaviors and nutritional status, both of which are linked to elevated blood pressure. Objective To examine the association between sociodemographic factors, subjective perception of health, lifestyle behaviors, anthropometric measurements, and cardiometabolic health markers with blood pressure in a sample of Brazilian adolescents. Methods A cross-sectional study was conducted with 273 adolescents (56% girls), aged 14–20 years, from Rio Branco, Acre, Amazon Region of Brazil. Data were collected using self-reported questionnaires and direct measurements, including anthropometric indices, cardiometabolic biomarkers, and blood pressure. Data were analyzed using chi-square tests and binary logistic regression models. Linear regression, adjusted by all variables, including dimension (sub-category) and age, was used to verify if the independent variables were associated with elevated blood pressure among adolescents. Results Significant associations were observed between elevated blood pressure and the male sex (OR = 2.56; 95% CI [1.11–5.86], p = 0.026) and rural residency (OR = 3.65; 95% CI [1.44–9.21], p = 0.006). Aerobic activity presented a significant association with elevated blood pressure (OR = 8.04; 95% CI [1.04–61.65], p = 0.045). Waist circumference increased the likelihood of elevated blood pressure (OR = 5.43; 95% CI [1.34–21.94], p = 0.018). Cardiometabolic markers, such as fasting glucose, demonstrated a significant association with elevated blood pressure (OR = 2.43; 95% CI [1.00–5.90], p = 0.048). No significant associations were found for muscle-strengthening activity, screen time, sleep duration, or food intake. Conclusion Our findings emphasize the crucial role of sociodemographic factors, especially the male sex and rural residency, in predicting elevated blood pressure among adolescents. Adolescents who perform the recommended amount of aerobic activity and who present acceptable fasting glucose values during adolescence are less likely to present high blood pressure. Waist circumference together with other anthropometric measurements might be a useful indicator for screening elevated blood pressure risk. These findings highlight the necessity for early detection, targeted interventions, and personalized strategies to reduce the hypertension risk and improve cardiometabolic health in the young population.
Nascimento-da-Silva et al. (2025) Association of sociodemographic factors, lifestyle behaviors, anthropometric measures, and cardiometabolic health markers with blood pressure in adolescents: a cross-sectional analysis. PeerJ 13:e20151 https://doi.org/10.7717/peerj.20151
Modulation of carbon-to-nitrogen ratio shapes the microbial ecology in a methanol-fed recirculating marine denitrifying reactor
Background Nitrate (NO3−) can accumulate in closed-circuit ecosystems to a toxic level. Adding heterotrophic denitrification process to the water treatment is a strategy to reduce this level. This type of process usually requires the addition of a carbon source. Carbon-to-nitrogen ratio (C/N) is a key parameter known to influence both the function and the activity of microbial communities in bioprocesses. Few studies have examined the influence of C/N on denitrification systems operated under methylotrophic and marine conditions. Here we assessed the influence of C/N (methanol and NO3−) on the performance of a laboratory-scale, recirculating denitrifying reactor operated under marine conditions. We monitored the evolution of the bacterial community in the biofilm to assess its stability during the operating conditions. Finally, the relative gene expression profiles of Methylophaga nitratireducenticrescens strain GP59, the main denitrifier in the denitrifying biofilm, were determined during the operating conditions and compared with those of GP59 planktonic pure cultures. Methodology A 500-mL methanol-fed recirculating denitrification reactor operated under marine conditions and colonized by a naturally occurring multispecies denitrifying biofilm was subjected to eight different C/N. We monitored several physico-chemical parameters (denitrifying activities, methanol consumption, CO2 production) throughout the operating conditions. The evolution of the bacterial community in the biofilm during these conditions was determined by 16S rRNA gene amplicon sequencing. Metatranscriptomes were derived from the biofilm to determine (1) the relative gene expression profiles of strain GP59, and (2) the functional diversity of the active microorganisms in the biofilm. Results Changes in C/N did not correlate with the denitrification dynamics (NO3− and NO2− reduction rates, NO2− and N2O dynamics), but did correlate with the methanol consumption rates, and the CO2 production rates. Throughout the operating conditions, nitrite and N2O appeared transiently, and ammonium was not observed. The bacterial community in the reactor increased in diversity with biofilm aging, especially among heterotrophic bacteria, at the expense of methylotrophic bacteria. The relative expression profiles of strain GP59 in the biofilm are distinct from those of planktonic pure cultures of strain GP59, and that the expression of several riboswitches and xoxF would be involved in these differences. Conclusions When the biofilm community is well established in the reactor, it can withstand changes in C/N with limited impact on the denitrification performance. The increase in the proportion of heterotrophs would allow the reactor to be more flexible regarding carbon sources. This knowledge can be useful for improving the efficiency of denitrification system treating close circuit systems such as marine recirculating aquaculture wastewater or seawater aquarium.
Lestin et al. (2025) Modulation of carbon-to-nitrogen ratio shapes the microbial ecology in a methanol-fed recirculating marine denitrifying reactor. PeerJ 13:e20129 https://doi.org/10.7717/peerj.20129