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Rebinding drives cellular retention of small-molecule drugs with abundant targets

Many small-molecule antitumor drugs are administered far less frequently than would be predicted from their plasma half-lives, in part because their efficacy is driven by prolonged retention within tumors. Drug retention can be caused by nonspecific protein binding, low membrane permeability, sequestration in acidic compartments, slow target dissociation, and rebinding to empty target sites, but the field has lacked a quantitative framework and preclinical assays for discriminating these mechanisms and predicting retention in vivo. We developed a theoretical framework and a simple two-drug assay for quantifying retention mechanisms in cell-based models. Application to 13 drugs with 8 distinct intracellular targets revealed diverse retention mechanisms. Rebinding emerged as a potent driver of retention whose magnitude was predicted by the target-to-affinity ratio ([Target]/ K d ). Rebinding slows the washout rates of important drugs such as taxanes and rapamycin by 100-fold or more. It also explains the need for high loading concentrations. Our approach bridges a critical gap in pharmacology modeling and provides practical tools to guide the design of intermittent dosing regimens.

Bayesian uncertainty quantification to identify population level vaccine hesitancy behaviours

When effective vaccines are available, vaccination programs are typically one of the best defences against the spread of an infectious disease. Such vaccination programs become particularly important during severe epidemics or pandemics to ensure sufficient vaccination coverage is achieved to increase protection or reduce transmission to a level that enables relaxation of non-pharmaceutical interventions, such as lockdowns, travel restrictions, or social distancing. Unfortunately, vaccination uptake in the community may be slow if there are substantial levels of vaccine hesitancy in the population. As a result, it is important to identify when these hesitancy behaviours are present in the community. Furthermore, understanding the main drivers of such behaviour can inform adjustments to public health strategies to improve community uptake. In this study, we consider the problem of identifying vaccination hesitancy behaviour during a vaccination roll-out that occurs in response to a severe epidemic. Specifically, our aim is to explore the extent to which mathematical modelling of reported case, death, and vaccination counts can be used to detect vaccine hesitancy and possible drivers. To do this, we develop a novel susceptible-exposed-infectious-recovered (SEIR) epidemiological model of disease transmission that incorporates changes in population behaviour relating to non-pharmaceutical interventions and vaccine uptake that are influenced by information reported through media or data dashboards about cases, deaths, and vaccination rates. We then use a Bayesian approach to analyse simulated data representing various hesitancy scenarios. Through this simulation study, our key findings are that individual parameters values related to drivers of vaccine hesitancy often cannot be identified. However, posterior correlation structures between these parameters enable the presence of vaccine hesitancy in the community to be detected and provide some insight into the relative influence of key factors, such as vaccine safety concerns or complacency. While our simulation study is inspired by the public health response to the COVID-19 pandemic, our tools and techniques are general and could enable vaccination programs of various infectious diseases to be adapted rapidly in response to community behaviours in the future.

Development of soil surface wetness models using machine learning techniques in the selected sites in Punjab, North-Western India

Abstract Accurate prediction of soil surface wetness (SSW) is vital for effective land management and resource optimization, particularly in sensitive ecosystems like the Western Himalayas. The main objective of the present study is to improve the accuracy of SSW prediction using hybrid and bagging models in the selected sites in Punjab, north-western India. The study utilizes ten machine-learning models comprising five base learners-random forest (RF), extreme gradient boosting (XGBoost), light gradient boosting machine (LightGBM), logistic model tree (LMT), and classification and regression tree (CART) and their corresponding AdaBoost-based hybrid variants: AdaBoost-RF, AdaBoost-XGBoost, AdaBoost-LightGBM, AdaBoost-LMT, and AdaBoost-CART. The SSW data set was collected from NASA POWER platform and Goddard Earth Observing System (GEOS) derived data which covers the period from 1986 to 2021. For model development, we applied a monthly data-lagging technique to generate different model scenarios. For feature selection, we applied greedy stepwise and best-first algorithms to identify the most effective predictors and improve model efficiency. Evaluations of the models were based on a variety of statistical indices. The results show that the RF model achieved the highest correlation coefficients (CC) across the study areas, ranging from 0.64 to 0.76 in Moga, 0.61 to 0.82 (XGBoost) in Hoshiarpur, and 0.41 to 0.64 (LMT) in Firozpur during the testing period. Accordingly, the hybrid AdaBoost-LightGBM model was the best, with CC values of 0.61–0.76, 0.63–0.82, and 0.49–0.60 for Moga, Hoshiarpur, and Firozpur, respectively. Overall model performance was limited to moderate and varied across locations and scenarios; although AdaBoost–LMT showed the best relative performance, the results primarily support its use for reproducing temporal variability in GEOS-derived SSW rather than precise wetness estimation. The findings contribute to improving SSW estimation and support data-driven decision-making for sustainable land and water management in the selected sites in Punjab, north-western India.

Combined generalist and host-specific transcriptional strategies enable host generalism in the fungal pathogen <i>Botrytis cinerea</i>

How generalist pathogens infect phylogenetically diverse hosts remains a central question in plant-pathogen biology. In particular, the extent to which broad host range is enabled by genetic variation vs. transcriptional plasticity is unclear. To investigate how variation and plasticity contribute to generalism, we studied the generalist necrotrophic fungus Botrytis cinerea that infects more than 1,500 plant species. Using a cross-infection matrix of 72 B. cinerea isolates infecting 57 plant genotypes across 15 eudicot species, we identified general and host-dependent fungal components of lesion formation. Transcriptome profiling at 48 h postinoculation revealed two distinct pathogen gene modules: 1) a set of general lesion-associated genes showing similar expression across hosts but varied among isolates; and 2) a set of high-entropy, host-inducible genes, organized into distinct coregulated modules that respond dynamically to specific host cues. Both gene sets were genomically dispersed, lacking structural clustering, and were under different levels of selective constraints. Our results demonstrate that B. cinerea employs a modular transcriptional strategy that integrates a core metabolic program along with a plastic, host-responsive regulatory network to achieve broad host colonization. This study presents the most comprehensive cross-species cotranscriptomic dataset to date for any fungal phytopathogen, highlighting transcriptional plasticity as a key mechanism underlying generalism in plant–fungal interactions. Moreover, the identification of conserved fungal gene targets across diverse hosts offers a foundation for developing broad-spectrum resistance strategies in multiple crops.

Transcriptomic and metabolomic analysis of allelopathic responses in Elymus nutans

To explore the allelopathic response mechanism of Elymus nutans to the extracts of Ligularia sagitta , seed germination tests combined with integrated transcriptome and metabolome analyses were performed to systematically investigate the effects of the extracts at different concentrations on its germination characteristics and molecular metabolism. The results showed that the concentration range of 0.01–0.10 mg/mL was the sensitive interval for the allelopathic response of Elymus nutans to the extracts of Ligularia sagitta , and germination energy (GE) exhibited higher sensitivity to allelopathic stress than germination rate (GR). Transcriptome analysis yielded a total of 367463 Unigenes. In the comparison group of A and B,5617 differentially expressed genes (DEGs) were identified, including 3122 down-regulated genes and 2495 up-regulated genes. GO enrichment analysis revealed that these DEGs were mainly involved in biological processes such as biosynthesis and structural constituents of ribosomes. KEGG analysis indicated that 2683 DEGs were mapped to 133 pathways, of which 14 were significantly enriched, mainly covering pathways related to secondary metabolite biosynthesis, ribosome, and plant hormone signal transduction. Metabolome analysis identified 361 differential metabolites (DMs), which were annotated to 79 pathways with 29 significantly enriched ones. Integrated transcriptome and metabolome analysis revealed that DEMs and DMs were mainly enriched in two KEGG functional categories, namely Genetic Information Processing and Metabolism. Among these, the phenylpropanoid biosynthesis pathway was identified as the core secondary metabolic pathway in response to allelopathic stress. This pathway relies on a series of enzymatic reactions catalyzed by key genes including phenylalanine ammonia-lyase ( PAL ), phenylalanine/tyrosine ammonia-lyase ( PTAL ), cinnamyl alcohol dehydrogenase ( CAD ) and cinnamoyl-CoA reductase ( CCR ), together with key metabolites such as p-Coumaric acid, Ferulic acid and Sinapyl alcohol, to produce metabolic products including lignin and flavonoids. This study provides a theoretical basis for revealing the molecular mechanism underlying the allelopathic response of Elymus nutans to the extracts of Ligularia sagitta , and also offers a scientific basis for the conservation and sustainable utilization of grassland ecosystems in alpine regions.

Development and evaluation of planar GSM antenna integra-tion into the housing of a navigation seal for transport and logistics

Abstract The article presents the results of a comparative study of 28 types of commercial flat GSM antennas and a self-developed antenna “Zmeyka” fabricated on a 1 mm FR-4 substrate operating in the GSM 900 band (Uplink 890 MHz, Downlink 960 MHz) for integration into the housing of a navigation seal intended for real transportation conditions. Particular attention is paid to the influence of housing materials, layout constraints, and the electromagnetic environment on impedance matching parameters and signal reception quality. The study was conducted in several stages. Laboratory measurements in an anechoic chamber using a Keysight FieldFox N9915A analyzer showed that in the reference housing made of 2 mm ABS plastic, the “Zmeyka” antenna achieved an average VSWR of 1.2, whereas several commercial samples exhibited VSWR values exceeding 6. Subsequently, selected antennas were tested in field conditions using a proprietary software tool “GSM Antenna Tester” to record CSQ/RSSI parameters. This approach enabled a comparative analysis of antenna performance not only under controlled laboratory conditions but also in scenarios close to real operating environments. Experimental results demonstrated that the integration of antennas into a housing made of reinforced polyamide PA12+GF ( $$\varepsilon = 3.5$$ –4.5) with 3.5 mm wall thickness and dense internal layout causes a downward shift of the resonance frequency by approximately 50 MHz, as well as an increase in VSWR up to 3.5 and a reduction of the transmission coefficient in the main lobe by 6–7 dB. Against this background, the self-developed “Zmeyka” antenna showed the highest resistance to layout and environmental effects, maintaining acceptable impedance matching and stable signal reception. Field tests using the “GSM Antenna Tester” software recorded a decrease in the signal level of commercial antennas to the “Weak” category (CSQ 7–11, RSSI from $$-99$$ to $$-91$$  dBm), while the adaptive design of the “Zmeyka” antenna compensated for environmental effects. The obtained results confirm that the design of navigation seals with integrated GSM communication cannot rely solely on the nominal antenna specifications provided by manufacturers. The electromagnetic environment inside the housing, material properties, and layout features have a significantly stronger impact on antenna performance than is typically assumed. It has been demonstrated that the possibility of adjusting the topology of the radiating elements in the “Zmeyka” antenna makes it possible to reduce insertion losses from 6–7 dB to an acceptable level of 1–2 dB, thereby ensuring stable data transmission under challenging transportation operating conditions.

TAZ integrates ECM remodeling with uterine stromal differentiation to maintain early pregnancy

The uterine microenvironment is critical for establishing pregnancy and sustaining embryonic development. Embryo attachment induces profound endometrial transformation, including stromal differentiation and vascularization, termed decidualization. This process, conserved in humans and rodents, supports the embryo even before placentation. Poorly formed decidua leads to fetal growth restriction or placental defects, yet underlying mechanisms remain unclear. Here, we used single-cell RNA-seq of mouse and human endometria, including patient samples, to uncover key regulators. We identified that collagen-related extracellular matrix (ECM) remodeling depends on TAZ, a central effector of Hippo signaling, and is essential for decidual formation. ECM pathways were enriched in differentiating stromal cells alongside upregulation of TAZ and associated transcription factors. Conditional uterine knockout of TAZ ( Wwtr1 flox/flox Pgr Cre/+ ; Wwtr1 -uKO) caused pregnancy failure in mice, marked by defective trophoblast invasion, early embryonic degradation, and severe subfertility. Spatial transcriptomic and histological analyses of TAZ-deficient endometria further revealed impaired decidualization, ECM remodeling, and vascularization. Thus, TAZ promotes temporal ECM remodeling at the feto–maternal interface, ensuring functional decidual zone formation and healthy pregnancy outcomes.

Ethanolic extract of Otostegia fruticosa induces ROS-dependent apoptosis and reduces migration of MDA-MB-231 cells in vitro

Breast cancer (BC) has a highly aggressive and metastatic nature. Specifically, triple-negative breast cancer (TNBC) is highly complicated to treat and recover from. Nowadays, treatment strategies focus on the beneficial nature of the herbal source. Otostegia fruticosa gained focus on treating various cancers due to its potential against cancer. This study aimed to investigate the apoptotic and metastasis properties of Otostegia fruticosa against triple-negative breast cancer cells (MDA-MB-231). The ethanol extract of Otostegia fruticosa exhibited cytotoxic effect on MDA-MB-231 cells, with an IC 50 value of 23 µg/mL. various tests, including MTT, morphological analysis, fluorescence staining, and Trypan blue exclusion, invasion, migration ability were employed to evaluate the effects of Otostegia fruticosa on cell viability and apoptosis. The results showed that Otostegia fruticosa induced apoptosis in MDA-MB-231 cells, characterized by the accumulation of reactive oxygen species (ROS), mitochondrial damage, and loss of nuclear potential. Furthermore, Otostegia fruticosa treatment reduced cell viability due to excessive ROS generation, as evidenced by DCFH-DA staining. Functional assays revealed that Otostegia fruticosa impaired cellular mobility, invasion, and colony-forming ability in MDA-MB-231 cells. The Otostegia fruticose significantly suppresses invasion property of breast cancer cells about 50% and significantly reduced migrated cells time dependent manner. Gene expression analysis by qRT-PCR and immunofluorescence showed altered expressions of genes linked to apoptotic (Caspase-3, 8, 9, and Bcl2, Bax, Bid, Cytochrome c, PTEN) and metastasis (MMP-9) protein. Notably, Otostegia fruticosa downregulated MMP9 and OPN protein expressions, while upregulating caspase-9 and Cyt-c. Western blot analysis validated these findings, highlighting the involvement of the p-Akt and OPN downregulation. Altogether, this study demonstrates that Otostegia fruticosa induces apoptosis and inhibits metastasis in MDA-MB-231 cells by regulating caspase-3, 8, and 9, Bcl2, Bax, and Bid. Meanwhile, it potentially arrests the translation level of MMP-9 and OPN proteins. In addition, cellular migration and invasion were significantly downregulated, and Otostegia fruticosa possesses the anti-tumor effect against the TNBC cells (MDA-MB-231).

Comparative health risk assessment of occupational dust exposure in industrial sectors using multiple models and monte carlo simulation

Migration-linked TB epidemics: How TB elimination in high-income countries depends on the success of TB control in high-burden settings

Mycobacterium tuberculosis is the leading global cause of infectious disease deaths. A growing proportion of tuberculosis cases in high-income countries occur among foreign-born individuals, due to infections acquired before migration. We developed a system of mathematical models representing tuberculosis epidemiology in 187 countries, linked by migration patterns, and trained on empirical data for each setting. Using these models, we quantified how future tuberculosis risks in high-income countries are influenced by the success or failure of efforts to combat tuberculosis in high-burden settings. Over 2025–2050, we projected 2270,000 (95%CI: 1930,000 to 2730,000) tuberculosis cases in high-income countries if pre-2025 trends continue, with 54% (46 to 62) among foreign-born individuals (versus 39% in 2024), for an average incidence of 6.0 (4.8 to 7.7) per 100,000 in 2050. Under the most optimistic scenario (assuming high-burden countries achieve aggressive WHO tuberculosis elimination targets), we estimated 667,000 (559,000 to 812,000) fewer tuberculosis cases in high-income countries over 2025–2050, with incidence of 2.6 (2.1 to 3.3) per 100,000 in 2050. Under the most pessimistic scenario (reduced international health aid causes rising tuberculosis in high-burden countries), we estimated 890,000 (793,000 to 998,000) additional tuberculosis cases in high-income countries, with incidence of 10.2 (8.4 to 12.7) per 100,000 in 2050. The United States, United Kingdom, Germany, Singapore, and France were the most affected high-income countries. For high-income countries, future tuberculosis incidence and mortality could vary by as much as 4 times depending on the success or failure of tuberculosis control in high-burden settings, fundamentally shaping strategies to prevent, detect, and treat tuberculosis in these settings.

A bioactive hydrogel integrating bFGF/VEGFA gene-loaded nanoparticles and platelet-rich plasma for accelerated full-thickness skin wound healing

Research objectives Full-thickness skin defects present substantial challenges to the healing process, arising from the loss of the dermal layer, inadequate vascular supply, and insufficient growth factor availability. Although previous studies have established that growth factors and platelet-rich plasma (PRP) individually promote tissue repair, the synergistic benefits of their combined application for achieving superior therapeutic outcomes remain unclear. Here, we introduce a novel composite biomaterial: a hydrogel integrating nanoparticles loaded with basic fibroblast growth factor (bFGF) and vascular endothelial growth factor A (VEGFA) genes (bFGF/VEGFA NPs) with PRP (termed bFGF/VEGFA@PRP hydrogel), engineered to enable the synergistic delivery of growth factors and bioactive components. Methods In this study, we hypothesized that the bFGF/VEGFA@PRP hydrogel can promote wound healing. This hypothesis was tested through various experimental perspectives and methods including material characterization, wound healing rate, wound blood flow signal intensity, skin attachment, cell proliferation, apoptosis, collagen deposition, and growth factor levels. Results In a rat cutaneous wound model, this hydrogel accelerated wound closure, enhanced local blood perfusion, and increased the density of skin appendages and collagen fibers. Mechanistic investigations revealed upregulated expression of angiogenic factors bFGF, VEGFA, and platelet and endothelial cell adhesion molecule 1 (CD31), the anti-apoptotic protein BCL2, the cell proliferation marker Ki67, and type III collagen (COL III) in the treated group. Conclusions Collectively, these results demonstrate that the bFGF/VEGFA@PRP hydrogel promotes wound healing through the synergistic enhancement of growth factor expression, angiogenesis, and COL III deposition. The combination of gene therapy with PRP treatment was found to be more effective than either modality alone. This study establishes a promising therapeutic strategy for managing complex full-thickness skin injuries.

Construction method for three-dimensional dynamic configuration of reciprocating sucker rod string in actual wellbore

Abstract The sucker rod string—serving as the core component of rod-pumped oil production systems—exhibits an ultra-slender geometric configuration. Under downhole reciprocating motion, it is prone to complex three-dimensional spatial deformation and may come into contact with the tubing wall, thereby inducing eccentric wear or, in severe cases, mechanical failure such as fracture. Since the dynamic spatial configuration of sucker rod strings is essential for understanding and mitigating tubing–rod eccentric wear, this study proposes a method to construct its three-dimensional spatial configuration under reciprocating movement. The method is grounded in a three-dimensional coupled dynamic model of the sucker rod string within tubing, incorporates buckling stability theory, and introduces smooth transition conditions that ensure deformation continuity of sucker rod string across distinct buckling modes. Finally, engineering case studies are employed to analyze the evolution of the sucker rod string’s three-dimensional spatial configuration within the tubing across distinct phases of reciprocating motion; moreover, the pivotal role of dynamic bottomhole pressure in triggering buckling mode transitions is elucidated.

Persuading large language models to comply with objectionable requests

Are large language models (LLMs) susceptible to the same persuasive appeals as humans? We tested whether classic persuasion principles (authority, commitment, liking, reciprocity, scarcity, social proof, and unity) could induce three widely used LLMs (GPT-5 mini, Claude Haiku 4.5, and Gemini 3 Flash) to comply with requests to assist with the synthesis of regulated substances. Across 126,000 conversations, persuasion principles increased compliance from 35.3% (at baseline) to 51.3% (using any principle). Although LLMs are not human, these findings underscore their parahuman (i.e., humanlike) nature and reveal the risk of manipulation by malicious users seeking to circumvent safety guardrails.

Correction: Reduced Retinal Microvascular Density, Improved Forepaw Reach, Comparative Microarray and Gene Set Enrichment Analysis with c-jun Targeting DNA Enzyme

HVD-Net: low-light image enhancement in HSV space via continuous hue encoding and dual-branch restoration

Pneumococcal membrane particles promote serotype-independent cellular and humoral immunity and protect against pneumococcal colonization

Despite implementation of pneumococcal conjugated vaccines (PCVs) against Streptococcus pneumoniae infections, these diseases are still major contributors to global mortality and morbidity, highlighting the need for a novel immunization strategy. Pneumococci release extracellular vesicles/membrane particles (MP) that are enriched in lipids, virulence factors, and surface proteins, and show great efficacy in protecting against invasive pneumococcal disease in mice. As such, MP present an alternative vaccine strategy. However, an in-depth understanding of how pneumococcal MP interact and modulate the immune system in vivo is currently lacking. In this study, we demonstrate that pneumococcal MP possess robust intrinsic adjuvant properties and conserved antigens, including MalX and PrsA, that support a strong serotype-independent effector and memory Th17 response in mice and humans. Mechanistically, vesicles are sensed through TLR2 and signaling through this receptor is required for the human IL-17 response. Moreover, intranasal immunization with MP expands the tissue resident macrophage populations in the lungs with enhanced functional capacities. Our study also demonstrates that MP immunization in the absence of adjuvant is sufficient to protect against colonization in mice.

Problematic cryptoasset trading is associated with greater depressive symptoms, anxiety symptoms, and social isolation

Public blockchains allow individuals to own, trade, and transfer digital cryptoassets on the internet. These cryptoassets are highly volatile and available to trade 24 hours a day, seven days a week. The potential financial rewards stemming from buying and selling cryptoassets create the possibility for individuals to display a maladaptive behavior, problematic cryptoasset trading, which can be considered as a type of problematic gambling. Prior research has demonstrated that behavioral addictive disorders, like gambling disorder, are associated with poor mental health. In addition, previous research has revealed that individuals who simply trade cryptoassets describe experiencing more negative mental health symptoms in comparison to non-investing individuals. Therefore, we hypothesized that problematic cryptoasset trading would be positively associated with three measures of mental health: depressive symptoms, anxiety symptoms, and social isolation. We collected cross-sectional survey data (N = 239; female = 78, male = 161) and ran hierarchical linear regressions, controlling for demographic characteristics. Our analyses supported the hypothesized associations between problematic cryptoasset trading and the three mental health measures. In other words, higher levels of problematic cryptoasset trading were associated with higher levels of depressive symptoms, anxiety symptoms, and social isolation. Implications for further research and clinical practice are discussed.

Driving mechanisms of tourist environmentally friendly behavior in cultural heritage destinations from an embodied cognition perspective

A BCG hydrogel enables localized NOD2/STING activation to overcome resistance to immune checkpoint blockade

Resistance to immune checkpoint blockade (ICB) often arises from immunologically cold tumors enriched in suppressive myeloid cells. Previous studies have implicated NOD2 signaling in antitumor immunity and in modulation of ICB responses, but approaches to engage this pathway effectively and durably within tumors remain limited. Here, single-cell transcriptomic analysis of colorectal cancer identified a NOD2 high tumor-associated macrophage (TAM) subset enriched for inflammatory and immune-activating programs. To therapeutically harness this state, we engineered an injectable manganese-containing alginate hydrogel encapsulating polyarginine-functionalized Bacillus Calmette-Guérin (MHY@PBCG) for sustained intratumoral delivery and localized coactivation of NOD2 and STING signaling in TAMs. Polyarginine enhanced BCG uptake by macrophages, whereas Mn 2+ stabilized the hydrogel and amplified STING activation. Local administration of MHY@PBCG reprogrammed TAMs toward an M1-like phenotype, increased inflammatory cytokine and interferon programs, converted cold tumors into immune-inflamed lesions, and restored responsiveness to anti-PD1 therapy in multiple models. Mechanistically, coordinated NOD2/STING activation established a self-reinforcing inflammatory circuit linking macrophage reprogramming to downstream T cell–mediated antitumor immunity. These findings establish a localized biomaterial strategy for overcoming checkpoint resistance through macrophage-centered remodeling of the tumor microenvironment.

Knowledge gaps and structural barriers to prescribing pre-exposure prophylaxis among healthcare providers in North Louisiana: A cross-sectional study

Background Pre-exposure prophylaxis (PrEP) uptake remains low in the Southern United States, despite disproportionate HIV burden. This study aimed to assess knowledge, attitudes, and structural barriers influencing PrEP prescribing among healthcare providers in North Louisiana. Methods We conducted a cross-sectional survey from January 2024 to April 2025 among 102 healthcare providers at a medical school and an affiliated hospital in North Louisiana. The survey assessed knowledge of PrEP, prescribing practices, and the perceived barriers. Descriptive statistics and multivariable logistic regression analysis were performed. Results Among 102 healthcare providers, only 27.5% had ever prescribed PrEP, and 48% had never discussed it with patients. The most frequently reported barriers to prescribing PrEP were lack of provider training (84.3%) and absence of clinical guidelines (68.6%). Multivariable analyses showed that providers with higher self-rated PrEP knowledge were significantly more likely to discuss PrEP with patients (adjusted odds ratio [aOR]: 5.10; 95% confidence interval [CI]: 2.31–11.27) and to prescribe PrEP (aOR: 3.29; 95% CI: 1.72–6.30). Conclusion Significant gaps in provider knowledge and institutional readiness persist in high-burden Southern settings. Targeted provider training and supportive institutional policies are essential to improving PrEP implementation.