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Numerical simulation study on pressure response and fluid loss mechanism of coal fracture network under particle temporary plugging

PLoS ONE Qiushuang Sun, Jian Chen, Liwen Guo et al. Jul 24, 2026 DOI: 10.1371/journal.pone.0352681

To address the issue of fluid loss leading to pressure failure when applying temporary plugging fracturing technology from the oil industry to enhance gas recovery in coal mines, and considering the current low efficiency of CO 2 solidification and utilization, a new method is proposed to inject SC-CO 2 and hot alkali solution together into coal to produce solidified particles as temporary plugging agents for solidification and utilization of CO 2 and fracturing transformation of coal. At present, the mechanisms governing pressure increase during temporary plugging with solidified CO 2 particles, as well as the relationship between internal pore and fracture pressures after plugging and fluid loss rate, remain unclear. Based on CT images, a complex geometric model of interconnected pores and fractures of coal is constructed. The morphology of solidified CO 2 particles is obtained via microscopic scanning, and conducts numerical simulation research on fluid solid coupling. Clearly define the pressure and flow distribution inside the coal under different temporary plugging positions and injection conditions. Revealing the pressure and fluid loss law inside the coal after temporary plugging. The results show that when particles are temporary plugging at different positions, the average internal pressure in the coal ranges from 8.05 MPa to 12.03 MPa. The fluid loss rate varies from 0.037 cm 3 /s to 0.152 cm 3 /s. Establish equations for the relationship between injection pressure, fracture size, temporary plugging location, internal pressure, and fluid loss rate, with a fitting formula R 2 of 0.988. Furthermore, an experimental setup and methodology involving supercritical CO 2 and hot alkali liquor injection were designed to validate the accuracy of the numerical simulation results regarding pressure enhancement and fluid loss under solidified particle plugging. This study provides theoretical support for enhanced coalbed methane recovery and the development of integrated technologies for underground CO 2 storage and hydraulic control of gas emissions.

Effects of transcutaneous auricular vagus nerve stimulation on depressive symptoms and executive function in post-stroke depression: a pilot randomized sham-controlled fNIRS study

Scientific Reports Yunjie Gui, Menghuan Wang, Yingying Ji et al. Jul 24, 2026 DOI: 10.1038/s41598-026-61616-1

Effects of different oral intake management during labor on maternal and neonatal outcomes: A protocol for a systematic review and network meta-analysis

PLoS ONE Zheng Nian, Yonghong Wang, Chuanya Huang et al. Jul 24, 2026 DOI: 10.1371/journal.pone.0354489

Introduction Fasting or restricting oral intake during labor may adversely affect maternal and neonatal outcomes. Although several interventions regarding oral intake during labor have been proposed, the optimal strategy for clinical practice remains controversial. This study aims to conduct a network meta-analysis to identify the most effective intervention for managing maternal oral intake during labor. Methods and analysis A comprehensive search will be conducted in the following electronic databases: Web of Science, PubMed, Embase, Ovid, Cochrane Library, ClinicalTrials.gov, the WHO International Clinical Trials Registry Platform, the Chinese Biomedical Literature Database, China National Knowledge Infrastructure, Wanfang Database, and VIP Database, from database inception until January 2026. All randomized controlled trials (RCTs) evaluating oral intake interventions during labor will be included. The primary outcomes include vaginal delivery rate, operative vaginal delivery, caesarean section, 10‐minute Apgar score, and neonatal hypoglycemia. Secondary outcomes include maternal and neonatal outcomes. Maternal outcomes include duration of labor, ketoacidosis, maternal hypoglycemia, postpartum hemorrhage, vomiting rate, Mendelson’s syndrome, maternal thirst and hunger sensations, and maternal satisfaction. Neonatal outcomes include fetal distress, neonatal jaundice, umbilical cord blood pH < 7.2, and admission to the neonatal intensive care unit (NICU). Study selection, data extraction, and quality assessment will be conducted independently by two reviewers, and disagreements will be resolved through discussion with a third reviewer. Meta-analysis will be performed using RevMan 5.4 where appropriate; otherwise, subgroup analyses or descriptive analyses will be conducted. Ethics and dissemination This review will not involve individual patient data and therefore does not require ethical approval. The findings of this systematic review and network meta-analysis will be disseminated through publication in a peer-reviewed journal and presentation at relevant academic conferences. PROSPERO registration number CRD42025630953.

Japanese tits discriminate between Eurasian sparrowhawks Accipiter nisus and Indian scops owls Otus bakkamoena

Scientific Reports Yu Zhang, Chao Shen, Tianxu Qiu et al. Jul 24, 2026 DOI: 10.1038/s41598-026-63515-x

Abstract Diurnal and nocturnal raptors exhibit significant differences in foraging times and predatory behavior, yet little is known about the nesting defense responses of parent birds to these raptors during the breeding season. We observed the behavioral response of Japanese tits Parus minor to taxidermy dummies of Eurasian sparrowhawks Accipiter nisus and Indian scops owls Otus bakkamoena . Our results show that the nest defense intensity of Japanese tits against sparrowhawks is significantly lower than that against owls. Specifically, Japanese tits predominantly engage in distant observation behavior when encountering sparrowhawks, whereas they display more aggressive behavior toward owls. Overall, we propose that Japanese tits discriminate between sparrowhawks and owls, with the latter posing a lesser threat to adult birds.

A multi-agent equilibrium model in an incomplete market with discrete dividends: Applications to long-term discount curves

PLoS ONE Taiga Saito, Akihiko Takahashi Jul 24, 2026 DOI: 10.1371/journal.pone.0343055

This paper develops a multi-agent equilibrium model in an incomplete market setting. The model incorporates dividend-paying securities whose dividend processes are interpreted as flows of consumption goods and can be driven by exogenously given factor processes. We consider an optimal consumption and portfolio problem for agents who have different views on fundamental risks and heterogeneous time preferences. Using a convex duality approach, we obtain expressions for the equilibrium state price density process, which subsequently yields the term structure of discount rates. To better reflect market practices, the model also incorporates discrete timing for dividend payments, consistent with semiannual or annual coupon schedules and policy decisions that typically occur at specific points during the year. As an application of the model, we provide numerical examples of long-term discount rates for valuing long-dated cashflows while exogenously incorporating the dynamics of factor processes that drive the dividend processes reflecting changes in the amount of government bonds available in the market. We examine how changes in the supply of government bonds affect the pricing of insurance products, including death benefits and pension annuities, through shifts in long-term discount rates. The numerical examples illustrate the qualitative implications of the model and are not intended to provide empirical findings.

Knowledge attitude and practice toward fertility preservation in patients with gynecologic cancers in Zhejiang of China

Scientific Reports Xiaoyan Fu, Wenqian Wang, Hanyu Shao et al. Jul 24, 2026 DOI: 10.1038/s41598-026-63450-x

Hidden deficiency under bright skies: Vitamin D prevalence and genetic associations in African Type 2 diabetes: A systematic review and meta-analysis

PLoS ONE Eyob Girma Abera, Samuel Alemu Himbaro, Surafel Worku Megersa et al. Jul 24, 2026 DOI: 10.1371/journal.pone.0354518

Background Type 2 diabetes mellitus (T2DM) has become a significant health challenge in Africa, with rapidly increasing prevalence. Despite abundant sunlight, vitamin D deficiency (VDD) is prevalent and may influence glucose metabolism and insulin resistance. Genetic factors, including vitamin D receptor (VDR) polymorphisms, may modify these effects. This systematic review and meta-analysis aimed to estimate the prevalence of VDD and evaluate the association of VDR gene variants with T2DM among African populations. Methods We systematically searched PubMed, Cochrane Library, ScienceDirect, Embase, Web of Science, and Google Scholar for observational studies published up to June 11, 2025, reporting VDD prevalence and VDR genetic associations in African individuals with T2DM. Data extraction and quality assessment followed Joanna Briggs Institute (JBI) and PRISMA 2020 guidelines. Pooled prevalence, mean difference and odds ratios (ORs) were calculated using random-effects meta-analysis. Heterogeneity, publication bias, sensitivity analyses, and meta-regression were performed. All statistical analyses were conducted using R version 4.3.1 (2023-06-16 ucrt). Results Twenty-two eligible studies encompassing 3,447 participants from 11 African countries were included. The pooled prevalence of VDD among individuals with T2DM was 51% (95% CI: 36–65%), with substantial heterogeneity (I 2  = 95.7%). Subgroup analyses showed highest prevalence in East Africa (56%). Vitamin D-deficient individuals had significantly higher HbA1c compared to sufficient individuals (mean difference = 0.89%; 95% CI: 0.06–1.72 p = 0.044; I 2  = 0%, p = 0.49). The pooled analysis of VDR FokI polymorphisms did not demonstrate a significant association with T2DM risk (OR = 1.51; 95% CI: 0.23–9.73). No evidence of publication bias was detected. Conclusion Vitamin D deficiency is highly prevalent in African populations with T2DM and is associated with poorer glycemic control. Current evidence is insufficient to conclude on VDR FokI polymorphisms and T2DM risk. These findings underscore the need for interventional trials to determine whether improving vitamin D status enhances glycemic outcomes; routine screening or supplementation recommendations remain premature. Registration number CRD420251134616.

Histology-validated quantitative MRI assessment of non-thermal focused ultrasound ablation

Scientific Reports Mingzhen Shao, Sumaiya Dipty, Wei Zhang et al. Jul 24, 2026 DOI: 10.1038/s41598-026-62578-0

Correction: Longitudinal changes in circulating biomarkers from baseline to week 48 in treatment-Naïve people living with HIV initiating integrase inhibitor-based antiretroviral therapy

PLoS ONE Jose-Ramon Blanco, Miguel Torralba, María Saumoy et al. Jul 24, 2026 DOI: 10.1371/journal.pone.0354685

Public awareness of pharmacovigilance and adverse drug reaction underreporting in Romania in a cross-sectional online survey

Scientific Reports Marius Călin Cherecheș, Diana Maria Olar, Răzvan Adrian Logigan Jul 24, 2026 DOI: 10.1038/s41598-026-63827-y

Randomized single-blind study to examine the implementation and effectiveness of integrating evidence-based early relational health programs in pediatrics for families with low incomes: A clinical trial protocol

PLoS ONE Caitlin F. Canfield, Elizabeth B. Miller Jul 24, 2026 DOI: 10.1371/journal.pone.0354239

Disparities in early childhood development due to poverty are linked to lifelong gaps in health and well-being. To address such disparities, interventions must not only include the prevention of toxic stress, but also the promotion of early relational health—that is the presence of safe, stable, nurturing relationships between young children and their caregivers. Moreover, a public health approach that integrates multiple programs in order to meet the unique needs and build on the varied resources of individual families is needed to effect population-level change. This randomized controlled trial seeks to examine the implementation and effectiveness of combining two evidence-based interventions, HealthySteps and PlayReadVIP. A total of 355 parent-child dyads, 35 in the implementation phase and 320 in the effectiveness phase, will be randomly assigned to receive HealthySteps+PlayReadVIP or traditional HealthySteps, as an active Control. Families will receive the intervention at each pediatric well-child visit from birth to age 2 and will complete assessments at enrollment and child age 12 and 24 months. In addition, up to ten HealthySteps Specialists and other care providers will be enrolled and will complete surveys at each assessment point. The primary outcomes include positive parent-child relationship, parent cognitive stimulation, and child socioemotional development.

Design, synthesis and biological evaluation of novel podophyllotoxin–melatonin heterodimers against ovarian cancer cell lines

Scientific Reports Pramukti Nawar Ra’idah, Zuzanna Molęda, Izabela Młynarczuk-Biały et al. Jul 24, 2026 DOI: 10.1038/s41598-026-62008-1

Tissue-specific amino acid accumulation in Cynomorium songaricum stem and epidermis responds to distinct water and soil cues

PLoS ONE Yu Tang, Xunchao Zhang, Yang Yang et al. Jul 24, 2026 DOI: 10.1371/journal.pone.0354162

This study provides the first evidence, from the perspective of intra-organ metabolic division of labour, that Cynomorium songaricum adapts to the heterogeneity of desert habitats through a synergistic strategy of “stem epidermis perceiving the water environment and fleshy stem buffering soil stress.” The research found a significant spatial metabolic division within the organs of C. songaricum : the stem epidermis is enriched in various essential and functional amino acids, including threonine and serine, with their content significantly higher than in the fleshy stem. In contrast, the fleshy stem specifically accumulates proline and cysteine, showing a significant difference. Environmental driving analysis indicates that, although C. songaricum is a holoparasitic root parasite, the metabolic division within its organs is still significantly driven by differential rhizosphere soil-water environmental factors: the amino acid profile of the stem epidermis is sensitive to water environmental factors such as electrical conductivity and carbonate ions, showing a positive correlation; whereas the amino acid composition of the fleshy stem is mainly regulated by soil factors such as water-soluble salts, showing a negative correlation. This suggests that, even while relying on the host for nutrition, the metabolic characteristics of C. songaricum are still influenced by its growth micro-environment. This finding breaks through the research paradigm of treating plants as homogeneous entities, providing new insights into understanding plant ecological adaptability and offering a key scientific basis for the artificial adaptive management and comprehensive utilization of C. songaricum resources.

Transparent deep learning and data-driven visual intelligence framework for robust crop recommendation in precision agriculture

Scientific Reports B. Dhiyanesh, Parveen Begam Abdul Kareem, P. Shanmugaraja et al. Jul 24, 2026 DOI: 10.1038/s41598-026-59598-1

Diagnostic accuracy of circulating microRNAs and lipidomic biomarkers for early breast cancer detection: A systematic review and meta-analysis

PLoS ONE Heba Mohammed Arafat, Akbar Ali, Tengku Ahmad Damitri Al Astani Tengku Din et al. Jul 24, 2026 DOI: 10.1371/journal.pone.0354728

Background Breast cancer outcomes improve substantially with earlier detection, yet mammography performance can be limited in dense breasts and may lead to false-positive investigations. Circulating microRNAs (miRNAs) and lipidomic/metabolomic signatures have emerged as promising minimally invasive biomarkers that could complement imaging for early-stage detection. The aim of this systematic review and meta-analysis is to evaluate the diagnostic accuracy of circulating microRNAs and lipidomic/metabolomic biomarkers for early breast cancer detection and to explore between-study heterogeneity. Materials and methods A PRISMA 2020–compliant systematic review and meta-analysis were conducted. MEDLINE (PubMed), Web of Science, Scopus, Springer, ScienceDirect, and the Cochrane Library were searched for eligible diagnostic studies assessing circulating microRNAs and lipidomic/metabolomic biomarkers measured in serum or plasma. No language restrictions were applied. Non-English reports were screened and, when potentially eligible, translated for full-text assessment and data extraction. Studies using whole blood were excluded. Study quality was assessed using QUADAS-2. Random-effects models (DerSimonian–Laird) pooled sensitivity, specificity, likelihood ratios, diagnostic odds ratio (DOR), and summary ROC (SROC) area under the curve (AUC). Heterogeneity was evaluated using Q and I 2 statistics, and small-study effects were assessed using Deeks’ test. Results Thirty-two studies (2015–2025) comprising 6,935 participants (3,697 breast cancer cases; 3,238 controls) were included. Pooled sensitivity was 0.87 (95% CI [0.83, 0.90]) and pooled specificity was 0.84 (95% CI [0.79, 0.88]), with pooled DOR 46.10 (95% CI [27.80, 76.70]), PLR 5.17 (95% CI [3.99, 6.68]), NLR 0.16 (95% CI [0.13, 0.21]), and SROC AUC 0.92 (95% CI [0.87, 0.94]). Heterogeneity was substantial (I 2  = 88.29% for sensitivity; I 2  = 90.20% for specificity). In subgroup analyses, serum-based studies showed higher pooled specificity than plasma-based studies. A formal threshold effect assessment did not reveal a statistically significant correlation between sensitivity and false-positive rate (Spearman ρ = − 0.334, p  =  0.062). Deeks’ test suggested potential small-study effects (p = 0.043). Conclusions Circulating microRNA and lipidomic/metabolomic biomarkers demonstrate strong overall diagnostic performance for breast cancer detection; however, substantial heterogeneity and potential small-study effects limit their immediate clinical translation. Given that most included studies used retrospective case-control designs rather than prospective screening cohorts, these biomarkers are best regarded as investigational, complementary tools rather than replacements for mammography at this stage. Future large, prospective, standardized studies with harmonized pre-analytics and prespecified thresholds are needed to support the implementation of screening or triage pathways.

A hybrid convolutional-transformer neural network model for photovoltaic fault detection and localization

Scientific Reports Ebrahim A. Ramadan, Nada M. Moawad, Belal A. Abouzalam et al. Jul 24, 2026 DOI: 10.1038/s41598-026-57859-7

Abstract Energy retention from losses is the primary goal of fault detection methodology for photovoltaic (PV) solar systems. A fault detection model should be designed effectively to minimize power and cost waste. We propose a novel fault detection and localization method that leverages deep learning techniques for PV systems. The model is a hybrid semantic segmentation method that combines Convolutional Neural Networks (CNN) and multi-level transformer neural networks. We examine our model on four different segmentation datasets, which have varied characteristics and different capturing conditions, to ensure our model generalization. Using aerial inspection, the first thermal dataset images give a high performance in locating the faulty cells in PV arrays with an accuracy and global precision of 99.89%, a mean average precision (mAP) of 88.73%, a mean intersection over union (mIoU) of 76.29%, and 84.48% of mean dice (mDice, or mean/unweighted F1 score). We use three electroluminescence datasets to investigate the precise location and class of different fault types and minor cracks at the cell level. The second dataset result achieved perfect detection and segmentation of anomaly areas in cells with 99% accuracy, 96.36% mIoU, 98.13% mDice, and 98.41% mAP. The first two datasets contain binary segmentation images with fault and no-fault classes; to accommodate multiple classes, we have utilized the third and fourth datasets. The third dataset, comprising five PV failure classes, is evaluated against other models, yielding a superior performance of 96.27% precision, 77.64% mAP, 57.3% mIoU, and 68.45% mDice. The final dataset has 29 segmentation classes for testing 25 classes, for which it achieves a precision of 95.05%, 66.7% mAP, 49.3% mIoU, and 59.3% mDice.

The news media and the agenda for noncommunicable diseases before and during the COVID-19 pandemic: Losing the competition for coverage and framing responsibility for action in Malawi

PLoS ONE Dereck L. Hamunakwadi, Stephanie L. Smith Jul 24, 2026 DOI: 10.1371/journal.pone.0341285

Chronic noncommunicable diseases (NCDs, such as cardiovascular and diabetes) pose an increasing burden in low-and middle-income countries, accounting for ~80% of premature (under age 70) NCD deaths globally. Despite their significant and rapidly growing burden—and status under United Nations Sustainable Development Goal 3.4—little is known about the agenda status of NCDs in lower-income countries. Though coverage of health issues shapes policymaker and public awareness and opinions, news media is an under-analyzed agenda setting arena. This study explores levels of priority for 10 NCDs compared to four infectious diseases and their burdens in the news media arena in Malawi; shifts in the agenda during the COVID-19 emergency; and media framing of actors responsible for addressing Malawi’s NCD burden. The study is informed by analysis of the contents of headlines (an indicator of media priority) and a purposefully selected subset of articles (framing analysis) published in two news outlets between 2015 and 2023. News media priority for NCDs in Malawi remained relatively low despite elevation to Sustainable Development Goal status. Between 2015 and 2019, news media priority in Malawi heavily favored HIV/AIDS, tuberculosis and malaria over 10 NCDs. Media priority shifted to COVID-19 and headlines on mental health surged ahead of those on HIV/AIDS and tuberculosis during the emergency (2020–2023). Alignment between the burden of disease and media priority was inconsistent. News media framed nongovernmental actors (primarily domestic) as responsible for addressing NCDs (predominantly cancers) far more frequently than other actors, including government, politicians and development partners. Featuring patterns of low attention and framing that fails to hold authorities responsible for addressing NCDs, media priority appears unlikely to substantially shift public opinion or policymaker agendas in their favor in Malawi. This study sets the stage for further analysis of media priority for health issues in lower-income countries.

The relative importance of resilience factors prior, during, and after stress-inducing exams in medical students

Scientific Reports Pascal Schlechter, Paul O. Wilkinson, Philip Jefferies et al. Jul 24, 2026 DOI: 10.1038/s41598-026-63690-x

Abstract Resilience is a dynamic process of adapting to stress. Identifying modifiable resilience factors (RFs) remains challenging due to the difficulty of studying individuals before, during, and after a stressor in real time. We addressed this by investigating modifiable protective factors in the context of university exams, a predictable and impactful stressor, and focused on medical students to examine RFs in a vulnerable cohort under standardized conditions. In this exploratory study, we used data from the three-wave ‘RESIST’ Study involving 451 medical students from the UK. Our investigation covered three timepoints with differing levels of stress (1) a non-exam period, (2) an exam period, and (3) post-exams at the end of the term. We examined the distinct impact of 13 modifiable RFs at individual, family, and community levels. Specifically, we investigated their relative importance and their cumulative impact with regard to self-reported mental health symptoms at three timepoints. We found the highest relative importance rankings for self-esteem, brooding, and distress tolerance. While most individual resilience factors had negligible-to-small effects, the sum of all 13 effects was similar to the effects of self-reported mental health symptoms at the previous timepoint on themselves. Our findings shed light on the role of RFs, and their cumulative and timing effects, and their associations with exam-related self-reported mental health problems among medical students.

Co-occurrence of antibiotic residues and Antimicrobial resistance genes in animal manure and agricultural soils from Machakos, Kiambu, and Kajiado Counties, Kenya

PLoS ONE Richard M. Machoka, Florence Ng’ong’a, Christabel Muhonja et al. Jul 24, 2026 DOI: 10.1371/journal.pone.0337311

The excessive and often unregulated use of antibiotics in livestock production and human health has led to the dissemination of antibiotic residues and antibiotic resistance genes (ARGs) posing a serious threat to the environment and public health. This study investigates the co-occurrence and spatial distribution of antibiotic residues and ARGs in livestock manure and agricultural soils from Machakos, Kiambu, and Kajiado counties in Kenya, regions characterized by intensive livestock farming. A total of 180 animal manure and soil samples from 30 farms across the three counties were collected and pooled into 18 samples. Antibiotic residues were extracted and quantified using high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Nine ARGs ( aadA, ermB , sul1, tetQ, tetW, dfrA1, blaMOX, blaOXA and qnrB were quantified via absolute and relative qPCR, normalized to 16S rRNA gene copy numbers. Multivariate analyses, including PCoA and Spearman Correlation, were conducted to explore ARG structure and co-occurrence. Tetracycline, particularly oxytetracycline, was the most abundant antibiotic in manure (up to 1150 ng mL −1 ), especially in pig manure from Kiambu. Sulfadimethoxine was undetectable in nearly all samples except in soil mixed with pig manure from Kajiado county, which showed a concentration above 70 ng mL −1 , the pig manure from the same county had levels below 2 ng mL −1 . Sulfadimidine, sulfamethoxanole and erythromycin were not detected in any of the samples. A three-way ANOVA showed that animal source (cattle, pigs, poultry) and sample type (manure, soil) were generally not significant factors for antibiotic concentration. Only oxytetracycline (P = 0.0421, 0.0901) and sulfadimethoxine (P = 0.0904, 0.044, 0.054) showed marginal significance by sample type (P = 0.0785). Antibiotic resistance genes were widely distributed, with aadA, ermB and sul1 being the most prevalent, especially in soils from Kiambu and Machakos. The tetQ showed extremely high relative abundance, indicating intense selection pressure from tetracyclines. Strong positive correlations were observed between co-occurring ARGs, including tetQ and tetW as well as erm and sul1 . The concurrent detection of persistent antibiotic residues and high ARG loads in both manure and soils underscores the urgent need for improved antibiotic stewardship, sustainable manure management, and environmental monitoring in Kenyan agroecosystems.

Expanded molecular pathomechanisms of GlyR α2 subunit variants in neurodevelopmental disorders and epileptic encephalopathy

Scientific Reports Sean D. Fraser, Anna-Lena Wiessler, Wing Yan Jessica Choi et al. Jul 24, 2026 DOI: 10.1038/s41598-026-61911-x

Abstract Rare genetic variants in the glycine receptor (GlyR) α2 subunit gene ( GLRA2 ) are associated with autism spectrum disorder, developmental delay, and intellectual disability, often accompanied by microcephaly, language delay or epilepsy. We report detailed structure-function analyses of nine previously uncharacterised GlyR α2 missense variants, including a novel de novo change (p.S285P) linked to epileptic encephalopathy. Using molecular modelling/dynamics simulations, electrophysiology, and immunocytochemistry, we assessed effects of GlyR α2 variants on agonist potency, efficacy, channel gating, and cell-surface trafficking. Five missense variants caused a partial loss-of-function via reduced glycine potency (p.F20S, p.A261T, p.R418Q), reduced glycine efficacy (p.F20S), or faster channel deactivation (p.R323C, p.P369T). By contrast, p.R225C abolished cell-surface expression resulting in a complete loss-of-function . The p.A261T variant also significantly reduced picrotoxin binding, resolving ambiguity in GlyR-PTX interaction models. Additional variants showed an alteration-of-function (p.I232M) or a gain-of-function (p.S285P), combining reduced glycine efficacy with increased potency and spontaneous leak currents. Two variants within the intracellular M3-M4 domain (p.R323C and p.P369T) had enhanced channel deactivation consistent with a loss-of-function , while p.P373L showed no detectable functional deficit. These findings expand the clinical and mechanistic spectrum of GlyR α2 variants, identify a key determinant of picrotoxin binding, and highlight unresolved roles of intracellular protein-protein interaction motifs in GlyR α2 function.