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Ahmed, S.

Publications and source records attributed to Ahmed, S..

At least 19 recordsLinked to original sources

A Framework for Clustering and Zero-Shot Labeling of Ecological Images Using Foundation Models

The increasing automation of biodiversity monitoring is generating large volumes of ecological images requiring taxonomic identification, creating a substantial annotation bottleneck. Assigning species-level labels requires considerable time and expert knowledge, limiting scalability. We propose a framework for unsupervised clustering and zero-shot labeling of ecological images using foundation models. The framework extracts image embeddings from pretrained models and groups images into taxonomic clusters without task-specific training. Species labels are assigned through zero-shot text matching between image embeddings and prompted information representing taxa expected in the dataset. We investigate dimensionality reduction and six clustering algorithms for organizing fine-grained imagery. An uncertainty score combines clustering and labeling information to triage images into three confidence tiers, allowing experts to focus on uncertain or potentially incorrect assignments. The framework is evaluated on four bird and moth datasets, including two annotated camera-trap collections. The camera-trap datasets, per-image species-level accuracy reached 84% on DK-Moth and 83% on HUGO-Bench Aves. An interactive report presents clusters and zero-shot labels by confidence tier and prioritizes cases requiring expert review. The framework provides a training-free approach for organizing and labeling ecological image collections, reducing manual annotation while retaining expert oversight and demonstrating the potential of foundation models for scalable, semi-automated annotation.

ecology↗

Sparse Phosphorylatable Residues, Tuned by Agonist Chemotype, Shapes GPCR Signaling

GPCR signaling diversity is often attributed to an intracellular phosphorylation barcode, in which ligand-dependent modification of multiple receptor phosphoresidues encodes selective engagement of downstream transducers. However, whether such an extensive phosphorylation of the receptor is universally required remains unclear. Here, we show that the melanocortin-4 receptor (MC4R), a central regulator of energy homeostasis, achieves remarkably diverse and pathway-selective signaling through a sparse set of strategically positioned regulatory phosphoresidues. Systematic study of these phosphoresidues within helix 8 and the C-terminal tail revealed distinct, spatially segregated contributions to receptor function. Substitution of helix 8 residues S306 disrupted beta arrestin2 recruitment, early endosomal trafficking, and ERK1/2 activation, while visibly altering G alpha-cAMP signaling in all the receptor variants. In contrast, distal C-terminal substitutions had little effect on beta-arrestin2 coupling and trafficking, revealing that these regions are not functionally redundant. Surprisingly, signaling through G alpha q/11- and G alpha12/13-linked pathways remained largely constrained across receptor variants, further demonstrating pathway-specific control by these phosphoresidues. Notably, ligand chemotype reshaped the functional consequences of these substitutions: setmelanotide selectively restored or enhanced signaling through pathways that were compromised by specific receptor mutations. Together, our findings argue that GPCR signaling diversity does not necessarily require an extensive phosphorylation barcode. Instead, MC4R uses a minimal number of spatially organized phosphoresidue that can differentially control transducer engagement and downstream signaling, while ligand chemistry determines how this regulatory landscape is accessed.

pharmacology and toxicology↗

Intracranial electrophysiology reveals distinct neural representations of nociception and salience in the human brain

Neuroimaging studies have shown close overlap between neural representations of nociceptive stimuli and non-nociceptive, salient stimuli (such as low-probability sounds), leading to the proposal that nociception and salience share a common neural substrate in the human brain. Here we use high-resolution intracranial recordings from 13 neurosurgical patients (2,018 total electrodes) to show that neural representations of nociception and salience are anatomically and functionally distinct in the human brain. 77% of 107 electrodes that encoded nociceptive stimuli did not encode low-probability sounds ("nociceptive representations"), whereas 23% encoded both nociceptive and low-probability sounds ("salience representations"). Nociceptive representations were distinct from salience representations in terms of anatomical prevalence (posterior insula, inferior parietal, and perirolandic cortex vs. anterior insula, thalamus, and inferior frontal gyrus), timing (nociception preceded salience by 120 ms on a trial-by-trial basis), and function (intracranial electrical stimulation was more likely to cause sensory experience when applied near nociceptive representations, as compared to salience representations). These results provide strong evidence that nociception and salience have distinct neural substrates in the human brain, and that selective measurement of these distinct processes may help understand patient-to-patient variability in chronic pain and develop personalized brain stimulation treatments.

neuroscience↗

Temozolomide-associated inflammatory-repair remodeling in recurrent glioblastoma exposes a ROCK-linked therapeutic vulnerability

Background: Glioblastoma (GBM) adapts to therapy through coordinated malignant-cell and microenvironmental responses, but the mechanisms linking treatment-associated inflammation to tumor-cell phenotypic plasticity remain poorly defined. Here, we investigated whether preoperative temozolomide exposure is associated with inflammatory-repair remodeling and a ROCK-linked therapeutic vulnerability in recurrent GBM using a window-of-opportunity clinical trial cohort. Methods: We integrated patient-resolved single-cell transcriptomics, spatial RNA profiling and multiplex protein imaging with experimental perturbations and orthotopic glioblastoma models. In the presurgical window-of-opportunity subgroup of NCT05236036, four patients with recurrent glioblastoma received additional preoperative temozolomide (TMZ; 150 mg/m2/day for 5 days) before resection and were compared with three recurrent comparators who did not receive additional preoperative TMZ. Results: Across seven tumors, malignant-cell inflammatory activity covaried with integrin-binding and wound-healing programs (median partial {rho} = 0.45 and 0.42, respectively). Tumors exposed to preoperative TMZ showed enrichment of inflammatory, chemokine, adhesion and wound-healing programs (GSEA q < 0.05), with the direction of enrichment preserved in all leave-one-patient-out analyses. In two tumors exposed to preoperative TMZ, CXCL12 localized to vascular territories, whereas chemokine and wound-repair programs increased near injury-reference regions. Across seven specimens analyzed by multiplex protein imaging, MYL9 abundance correlated with local CXCL12 ({rho} = 0.61), phosphorylated MYPT1 ({rho} = 0.48) and nuclear phosphorylated STAT3 ({rho} = 0.45), with positive associations in every specimen. Experimentally, TMZ increased CXCL8 and reactive-state markers and enhanced subsequent scratch closure, while CXCL8 and CXCL12 increased MLC2 phosphorylation. Fasudil attenuated TNF-NF-{kappa}B, inflammatory-response and IFN-{gamma}-response programs in TMZ-treated cells and reduced scratch closure and p-MLC2 in complementary assays. Across three orthotopic models, fasudil plus TMZ prolonged survival versus TMZ alone (model-stratified HR, 0.21; 95% CI, 0.07-0.61; P = 0.001). Conclusions: These findings link treatment-associated inflammatory-repair programs with cytoskeletal remodeling and support further translational evaluation of fasudil plus TMZ in glioblastoma.

cancer biology↗

From Predicted Ki to Surrogate IC50: Similarity-Guided Empirical Calibration of Drug Target Affinity Predictions

Drug target affinity models return the endpoint on which they are trained, whereas medicinal chemistry decisions are often made with a different assay readout. Here, we trained a DeepPurpose model to estimate inhibition constants (Ki) from molecular graphs and protein sequences and asked whether those predictions could be aligned empirically with measured IC50 values without treating Ki and IC50 as interchangeable. A BindingDB-trained checkpoint retained useful cross-target ranking on the Davis kinase benchmark without Davis training data (concordance index 0.860). We then rebuilt the Ki training set from ChEMBL 37 records coded as Binding assays (assay_type = 'B'), after removing censored records and targets with poor replicate reproducibility. This reduced median fold error from 16.35x to 9.23x on a leakage-cleaned kinase panel and from 15.30x to 7.90x on a 14-target non-kinase panel before any IC50 calibration. Similarity-guided leave-one-out calibration further reduced the non-kinase panel median error to 3.12x for the original checkpoint and 3.22x for the ChEMBL checkpoint at Tanimoto T = 0.6. Because retraining removed a substantial part of the apparent correction, we interpret the calibration as a target- and chemistry-dependent empirical offset between model output and IC50 assay space, not as a mechanistic Ki-to-IC50 conversion. In a separate project-level stratification of public records, median replicate variability was 2.41x for the subset classified as biochemical Ki and 3.30x for biochemical IC50; a small-sample correction placed the Ki variability nearer 2.8x. These values provide an empirical scale for the remaining calibration error rather than a theoretical performance limit. Similarity, rather than the number of calibrators, governed the main accuracy coverage trade-off. The resulting values are surrogate IC50 estimates for cross-target triage; within-target ranking remains a limitation of the present architecture.

biochemistry↗

Genomic Insights into Multi-Drug-Resistant, Klebsiella pneumoniae Isolated from an Infertile Woman: A Comprehensive Analysis of Antibiotic Resistance and Virulence Factors

Objectives: Klebsiella pneumoniae is a significant global health concern due to its ability to cause diverse infections and its increasing multidrug resistance. This study analyzed the whole-genome sequence of multidrug-resistant K. pneumoniae strain CP4, isolated from a vaginal swab of an infertile woman, to characterize its virulence factors, antibiotic resistance mechanisms, and epidemiological features. Methods: The Kirby-Bauer disk diffusion assay was used to screen antibiotic resistance. Whole genome sequencing was conducted using the Illumina MiSeq platform and followed by comprehensive bioinformatic analysis. Results: Antibiotic susceptibility testing revealed that the K. pneumoniae strain CP4 exhibited resistance to 11 of 18 tested antibiotics, including carbapenems, and displayed a Multiple Antibiotic Resistance (MAR) index of 0.611. Whole-genome sequencing identified an array of putative virulence and metal-resistant genes, classifying it as a human pathogen with a probability score of 0.887. The strain was characterized as sequence type 327 with K16 capsular and O1/O2v1 somatic antigens. The antibiotic resistome included 38 putative genes conferring resistance to a spectrum of antibiotics. Moreover, point mutations were identified in several genes that ResFinder predicted to be associated with resistance to fluoroquinolones, cephalosporins, and carbapenems. Additionally, K. pneumoniae strain CP4 harbored six putative plasmid-replicon types and 454 putative mobile genetic element-associated protein hits among predicted ORFs. Several putative antibiotic resistance genes, including tet(A), floR, mph(A), and aph(6)-Id, were associated with insertion sequences, transposons, and integrons, indicating a potential for horizontal gene transfer. Conclusions: The genomic features of multidrug-resistant K. pneumoniae CP4 highlight its potential for antimicrobial resistance and virulence, emphasizing the need for continued genomic surveillance and targeted interventions. Keywords: Klebsiella pneumoniae, Whole Genome Sequencing, Antimicrobial Resistance, Pangenome analysis

microbiology↗

PMPNN-DDG: an accurate machine learning-based {triangleup}{triangleup}G prediction pipeline trained on a novel interpretable feature set extracted from ProteinMPNN

An accurate and tractable approximation of the single-point mutation-induced change in protein thermodynamic stability, denoted by DDG, is critical for understanding the genotype-phenotype relationship. Several computational methods have been proposed for this problem; however, limited and error-prone training data and the difficult-to-predict magnitude of structural perturbations make this a challenging task. Consequently, the computational predictors proposed throughout the past decade incrementally improved prediction performance by proposing novel features, combining existing features, task-adapted neural network architectures, loss functions, data augmentation techniques, and pre-training procedures. In this work, we propose PMPNN-DDG, a Random Forest-based DDG prediction model, trained on a novel set of interpretable features extracted from the recently proposed message-passing neural network-based fixed backbone protein design model, ProteinMPNN. On the S669 independent test set, PMPNN-DDG achieves rF +R = 0.64 and RMSE = 1.45, outperforming all compared baseline methods across the reported evaluation measures. On the Ssym independent test set, it achieves rF +R = 0.81, rF -R = -0.99, and RMSE = 1.10, showing competitive performance relative to the compared baselines. PMPNN-DDG is publicly available at https://github.com/dRanger666/PMPNN-DDG.

bioinformatics↗

Benchmarking single-cell foundation models in a zero-shot setting

Single-cell foundation models have recently emerged as a promising approach for learning general- purpose representations from large-scale transcriptomic data. These models are trained on millions of cells and are designed to transfer their learned representations to a wide range of downstream tasks. However, their practical benefits compared to traditional approaches are still not fully understood. This study evaluates four foundation models, namely scGPT, SCimilarity, UCE, and Transcriptformer, across four downstream tasks: cell type annotation, human data integration, cross-species data integration, and protein expression prediction. Embeddings generated by each model were assessed using multiple public single-cell datasets and compared against conventional machine learning baselines. Performance was measured using task-specific evaluation metrics, including classification, integration, and regression metrics. The results showed that foundation model embeddings did not consistently outperform traditional approaches. In the cell type annotation task, baseline methods achieved the strongest performance across most datasets. For protein expression prediction, however, embeddings from the foundation models generally produced more accurate predictions than the baseline, with SCimilarity achieving the lowest prediction error and Transcriptformer obtaining the highest correlation scores. In the data integration task, all foundation models produced moderate results, while scVI (the baseline) achieved the strongest integration performance. Overall, the results suggest that current single-cell foundation models provide useful representations for some downstream tasks in zero-shot conditions but do not yet offer a universal replacement for task-specific methods. Their effectiveness remains dependent on the application and evaluation setting.

bioinformatics↗

High-fidelity rare structural variant detection with HiFiRE3 reduced representation via restriction enzyme ends

High-fidelity detection of rare structural variants (SVs) remains challenging because library preparation and sequencing techniques generate artifactual junctions that obscure true single-molecule events. Here, we present HiFiRe3, an error-minimized sequencing framework that combines artifact-aware library design with error suppression and correction strategies to enable rare SV detection and frequency assessment across long and short-read sequencing platforms. We first systematically characterized major classes of SV artifacts, including chimeric PCR products, intermolecular ligation, sequencing platform-specific artifacts, and mapping errors. HiFiRe3 supports error correction of these artifact junctions by combining reduced representation restriction fragments with pre-ligation size selection to enable computational filtering via independent forced restriction enzyme end (FREE) and <1N size logics. In nanopore libraries, these approaches enabled targeted detection of single-molecule SVs at replication stress hotspots in cultured human cells exposed to genotoxicants and in long genes in untreated mouse brains, while markedly reducing singleton translocation artifacts. HiFiRe3 extends to PacBio sequencing for joint SV and SNV error correction and to short-read platforms for cost-efficient high-fidelity nonhomologous SV analysis. Together, HiFiRe3 is a flexible framework for accurately detecting rare genomic structural variation with broad applicability to targeted and genome-wide studies by selective application of its error correction approaches.

genomics↗

Multidomain triple target capture for navigating complex symbioses

Amplicon sequencing remains the benchmark technique for characterizing microbial communities, but the limitations of PCR bias and single-locus targets conspire to limit conclusions. Metagenomics, the primary alternative, shares with amplicon sequencing challenges of economic scaling at adequate sequencing depth. Targeted enrichment strategies can improve the data resolution and economics of sequencing efforts while reducing methodological bias. Here, we develop a target capture strategy for metagenomic characterization of eukaryotic ribosomal DNA and root nodule symbiosis genes and test it in the metagenomes of plant root nodules. We utilize biotinylated RNA probes to selectively capture genomic regions of interest from complex environmental DNA samples, avoiding forms of PCR bias that can undermine community characterization and overcoming the need for conserved priming sites often lacking in functional genes. We designed custom probe sets targeting conserved flanking regions of eukaryotic ITS and known root nodule symbiosis (RNS)-related genes and tested them on diverse root nodule metagenomes and a mock community. We observed high recovery of target loci from samples, very high on-target read proportions, and enhanced detection of low-abundance taxa compared to amplicon sequencing, including stronger alignment with known mock community compositions. This approach will enable deeper insights into the phylogenetic diversity of eukaryotic symbionts, their genomic adaptations, and the functional potential of symbiotic interactions in a cost-effective manner suitable for large-scale projects. This strategy advances our understanding of microbial community dynamics and symbiotic relationships in natural and anthropogenic ecosystems.

ecology↗

"Fuzzy specificity" shapes diazotroph diversity and composition in nodulating plants of the Southeastern USA

Nitrogen-fixing symbioses, particularly those occurring in root nodules, are among the most consequential mutualisms in natural and agricultural systems and represent a globally important source of bioavailable nitrogen. Despite their importance, patterns of diversity and composition among diazotrophic symbionts--and the processes structuring those patterns in natural systems--remain poorly resolved, with competing hypotheses emphasizing ecological, or phylogenetic constraints on host-symbiont associations. Here, using a broad survey of nodulating plants from the southeastern United States, we examine how diazotrophic symbiont communities vary across host plant phylogeny, habitat context, and geographic origin. We find that host phylogeny is the primary determinant of symbiont composition, outweighing effects of fine-scale taxonomic identity. Symbiont associations are therefore structured mainly at deeper phylogenetic levels, consistent with phylogenetically constrained, or "fuzzy," host specificity. Likewise, nodule community diversity--potentially reflecting variation in host control over infection--differs primarily among higher-level clades rather than among closely related taxa. Habitat context also shapes nodule communities, but its influence is secondary and most evident in undisturbed environments. As well, nonnative legumes harbor distinct symbiont assemblages despite occupying similar habitats, whereas distantly related legume clades share symbionts across habitats, highlighting interactions among phylogeny, ecology, and geographic history. Overall, our results show that host phylogeny exerts the strongest influence on nodule microbial communities, likely reflecting evolutionary divergence in symbiotic function across major host lineages.

ecology↗

Differences between males and females in psychostimulant/sucrose self-administration (when observed) may not necessarily be driven by biological sex.

BackgroundSex differences in psychostimulant-related behaviors are often attributed to biological sex; however, individual variability may also strongly influence behavioral outcomes. The new MISSING (Mapping Intrinsic Sex Similarities as an Integral quality of Normalized Groups) model identifies mixed-sex behavioral groups in which differences are driven primarily by individual variability rather than sex. The goal of this study was to validate the MISSING model for psychostimulant/sucrose self-administration. MethodsLong Evans rats self-administered methamphetamine (METH, male n = 25, female n = 32, 0.1 mg/kg/infusion, FR1, 6h per day for 20 days), sucrose (male n = 20, female n = 22, one-20 mg pellet/delivery, all other conditions being equal) and saline (male n = 3, female n = 10, other things being equal). We developed a new Quantitative Structure of Curve Analytical (QSCAn) model (using exponential-plateau and linear fit) for the assessment of individual drug self-administration time curve profiles irrespective of biological sex. We analyzed our data using regression analysis and ANOVA. ResultsQSCAn identified three distinct self-administration profiles (consisting of both sexes), which we named exponential-plateau negative (EP-), exponential-plateau positive (EP+), and undefined (EP0). There were no differences in self-administration profiles when we compared males and females within the same group. Differences between sexes (when observed) were due to mismatched comparisons (males from one group versus females from a different group). ConclusionsOur study reinforces the MISSING model for psychostimulant and sucrose self-administration by indicating that differences between males and females (when observed) may not necessarily be driven by biological sex. Significance StatementCurrent approaches often interpret variability in psychostimulant self-administration between males and females primarily through the lens of biological sex. However, this framework may overlook meaningful behavioral phenotypes shared across sexes. The present quantitative model suggests that individual patterns of behavior may better account for variability than sex alone, particularly in behaviors not strongly driven by sex-hormone-dependent mechanisms such as drug self-administration. By classifying animals according to behavioral profiles rather than biological sex, this approach may foster the identification of clinically and biologically relevant phenotypes underlying psychostimulant reinforcement.

neuroscience↗

Tau Disaggregation by a CNS-Permeable Small Molecule Reduces Fibril and Oligomer Burden and Preserves Proteostasis and Behavior

Pathological tau aggregates drive neuronal dysfunction in Alzheimer's disease (AD) and related tauopathies, yet no approved therapy eliminates existing tau neurofibrillary tangles. Here, we report the development of a coumarin-based small-molecule series that disaggregates tau fibrils and oligomers through a stacking-driven co-assembly mechanism. Structure-activity relationships identified PT-13 as a lead compound that inhibits tau seeding by AD brain-derived matter and reduces aggregate burden measured across both fibrillar and oligomeric tau species. Mechanistic studies demonstrate that disaggregation does not generate soluble oligomeric intermediates, addressing a central question in the field. PT-13 is brain-penetrant and well tolerated in vivo. In a tauopathy mouse model, PT-13 treatment reduces tau pathology while preserving behavioral function, proteasome capacity, and synaptic integrity. These findings establish small-molecule tau disaggregation as a viable therapeutic strategy and provide a molecular framework for the design of aggregate-directed therapeutics in neurodegeneration.

neuroscience↗

The recreational-to-habitual shift in psychostimulant use is an economic demand parameter that is unrelated to drug consumption levels (under normal and punishment conditions).

RationaleThe progression of psychostimulant abuse is associated with a shift from recreational to habitual use (R2H-shift). Because this R2H-shift can be modeled using behavioral economics, we developed a novel Behavioral Economic model for the Analysis of Self-administration Time-curve (BEAST) to obtain R2H-shift variable(s). The relationship(s) between R2H-shift variables and drug intake (under normal and/or punishment conditions) is/are unknown. Our goal was to determine if the R2H-shift variable and intake variables obtained during the initial self-administration training phase were related to 1) drug intake at that time, and subsequent drug intake under 2) normal, 3) punishment, 4) post-punishment, and 5) price-constrained conditions. MethodLong Evans rats self-administered methamphetamine (METH, males n = 16, females n = 14), sucrose (males n = 22, females n = 22) and/or saline (males n = 3, females n = 10) under FR1 for 6 h per day for 20 days to obtain 1) followed by the assessment of subsequent drug intake under different conditions (2-5 above). We obtained all variables referenced above. We determined the relationships between all variables (multivariate analysis). ResultsThere were no sex differences detected in the METH and sucrose studies. For METH and sucrose, prior drug intake levels could predict drug intake under normal/punishment but not under price-constrained conditions. The R2H-shift variable could predict drug intake under a consumption-price curve but could not predict intake under normal/punishment conditions. ConclusionsWhile related to economic demand, the recreational-to-habitual shift rate was unrelated to drug intake levels (under normal and punishment conditions).

neuroscience↗

Structural Characterization of Calcium-Dependent Calmodulin-Calmidazolium Binding using Capillary Vibrating Sharp-Edge Spray-based Native Mass Spectrometry and In-Droplet Hydrogen Deuterium Exchange Mass Spectrometry

Capillary vibrating sharp-edge spray ionization (cVSSI)-based native mass spectrometry (nMS) and in-droplet hydrogen deuterium exchange (HDX-MS) were used to evaluate calcium-dependent interactions between calmodulin and calmidazolium (CDZ). cVSSI was found to produce a narrow charge-state distribution (CSD) with low average charge states, indicating that this method preserved native-like states. cVSSI was also able to resolve stepwise Ca2+ binding for species containing one to four Ca2+ ions. In the absence of Ca2+, no detectable CDZ binding was observed. However, CDZ binding was observed when calmodulin was fully loaded with Ca2+. CDZ binding to the protein caused marked redistribution of the CSD toward lower charge states, consistent with ligand-induced stabilization of the protein into a more compact conformational ensemble. The apparent dissociation constant (Kd) of the interaction was determined to be 261 {+/-} 29 nM and 126 {+/-} 17 nM from Langmuir and quadratic binding models, respectively. Complementary in-droplet HDX-MS revealed that deuterium uptake was correlated with charge state across the CSD, corroborating the nMS evidence for stepwise conformational compaction of CaM. Within the individual charge states, two distinct populations representing CDZ-bound and unbound CaM were resolved by HDX-MS, which were not detectable by nMS alone. Comparison of these populations within each of the 6+, 5+ and 4+ charge states revealed reductions in deuterium uptake by approximately 37%, 22%, and 11% upon CDZ binding, respectively, consistent with ligand occlusion and local structural tightening at the binding interface. The gradient in protection across charge states further reflects differences in binding interface accessibility among CaM conformers spanning a range of CDZ-induced conformational states. Together, these results demonstrate that cVSSI-based nMS coupled with in-droplet HDX-MS provides an integrated platform for simultaneously resolving metal loading, ligand binding, binding affinity, and ligand-induced conformational changes. This approach complements traditional structural methods by enabling direct interrogation of dynamic, metal-dependent protein-ligand interactions in their native states.

biochemistry↗

Predicting Antibody Self-Association with Sequence Structure Fusion Models: The Central Role of CSI-BLI in Early Developability Screening

Antibody-based biologics are expanding rapidly, yet challenges in development from self-association, high viscosity, aggregation, and unfavorable clearance underscore the need for accurate in silico screening. Clone self-interaction biolayer interferometry (CSI-BLI) is a plate-based, low-material assay of weak, reversible self-association that serves as an early proxy for high-concentration viscosity and a complementary predictor of in vivo clearance. In a 246-mAb panel, CSI-BLI moderately correlates with viscosity; further, in hFcRn Tg32 mice (41 antibodies), CSI-BLI strongly associates with clearance. Here, we present an end-to-end framework that distinguishes high versus low self-interacting clones (CSI-BLI class) by coupling a fine-tuned protein language model (ESM-2) with residue-aligned 3D context from AlphaFold-predicted structures encoded as residue graphs. Disentangled multi-stream attention fuses sequence content, chain-aware positional information, and structural signals to capture spatially proximate interactions that are distant in sequence. Edit-distance-controlled splits across 1499 IgGs and 988 VHHs assess generalization. The structure-aware model achieves the highest hold-out performance (VHH-Fc F1 = 0.76; IgG F1 = 0.57), while a sequence-only disentangled variant outperforms a standard PLM baseline without structural inputs. Complementary biophysical feature-based models, built from AlphaFold structures and sequence/structure-derived physicochemical descriptors with cluster-aware selection, deliver robust, interpretable performance (VHH F1 = 0.72; IgG F1 = 0.57), with SHAP analyses highlighting charge/dipole, hydrophobicity, and aggregation-propensity drivers across CDRs and frameworks. This interaction-aware sequence-structure framework, supported by interpretable feature models, is extensible to other developability endpoints and broader protein classification tasks where joint modeling of language-derived representations and residue-level geometry is advantageous.

bioinformatics↗

Structure and mechanism of the human TMEM260 O-mannosyltransferase

Protein O-linked mannose (O-Man) glycosylation is essential for mammalian development, and mutations in its biosynthetic glycosyltransferases cause severe muscular, neurological and cardiac disorders. Despite its biological and clinical importance, the structural basis of mammalian O-Man biosynthesis has remained unknown. Here we report cryo-electron microscopy (cryo-EM) structures of human TMEM260, an endoplasmic reticulum glycosyltransferase that selectively catalyzes O-mannosylation of semaphorin plexin receptors and receptor tyrosine kinases cMET and RON, key regulators of cell guidance and migration. Structures of TMEM260 in a ternary complex with its natural donor dolichyl-phosphate-{beta}-mannose (Dol-P-Man) and an acceptor peptide derived from plexin-B2, together with binary complexes with Dol-P-Man or a synthetic donor analogue, capture physiologically relevant, substrate-loaded states and reveal the structural basis of O-Man transfer. We identify a conserved O-mannosylation sequon that underlies acceptor specificity and show that TMEM260 modifies extended polypeptide substrates, consistent with a co-translational glycosylation mechanism. These findings establish the molecular mechanism of a mammalian O-mannosyltransferase required for the maturation of physiologically critical receptors and provide a structural framework for interpreting TMEM260-associated congenital malformations.

biophysics↗

Omics-analyses of Fermented Onion pickle in Shaping Gut Microbiota and Immune Response in Women: A Community-Based Trial in Pakistan

A fermented-food intervention trial conducted in Pakistan suggested beneficial changes in the composition of the gut microbiota in healthy women. Using a subset (n=17) of the same participants, this study further investigates the impact of fermented food (onion pickle) on gene expression levels using RNA transcriptomics, with a focus on host-microbiome interactions. After consuming pickles (50g/day) for eight weeks, blood and stool samples of participants were collected at baseline and post-intervention to assess inflammatory markers, 16S rRNA gene sequencing, clinical parameters, and RNA sequencing. Among inflammatory biomarkers, lipocalin (LCN-2) levels significantly decreased (pre=86.5{+/-}80.1ng/mL, post=61.0{+/-}59.0 ng/mL, p=0.04, paired T-test). Additionally, the intervention downregulated pathways (p<0.05) involved host responses to microbial stimuli, including response to bacterial origin, chemotaxis, and response to lipopolysaccharide. In gut microbiota, observed -diversity significantly increased post-intervention (p=0.02). Linear discriminant analysis effect size (LEfSe) revealed differential expressions (LDA [&ge;] 2.0) of Olsenella and Coriobacteriales at week-8, where Olsenella sp. showed a significant negative correlation with LCN-2 (R=-0.36, p<0.05, Spearmans correlation). These findings suggest that fermented onion pickle consumption for eight weeks modestly alters gut microbial diversity and composition and is associated with reduced inflammatory markers and altered host immune-related gene expression, potentially improving intestinal health. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=81 SRC="FIGDIR/small/711246v1_ufig1.gif" ALT="Figure 1"> View larger version (27K): org.highwire.dtl.DTLVardef@18679e0org.highwire.dtl.DTLVardef@1ab62eeorg.highwire.dtl.DTLVardef@ec318forg.highwire.dtl.DTLVardef@a9b94c_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗