Biology subjects

Raut, S.

Publications and source records attributed to Raut, S..

7 recordsLinked to original sources

bioRxiv2026

Glycocalyx-Directed Enzymatic Hydrogels Unlocks Fibroblast Regeneration to Promote Diabetic Wound Healing

Chronic diabetic wounds remain a major clinical challenge because current therapies address infection or supplement growth factors without correcting the cellular dysfunction that prevents regeneration. We identify pathological glycocalyx thickening in diabetic dermal fibroblasts (DDFs) as a driver of elevated caspase 3, 8, and 9 expression and heightened apoptosis -- deficits that stall wound repair. Cleavage of sialic acid residues by neuraminidase (NMase) reverses this dysfunction, restoring fibroblast migration, proliferation, and contractility. We engineer a photo-crosslinked hybrid hydrogel combining methacrylated gelatin (GelMA) with high-molecular-weight methacrylated chitosan (HMW ChMA). ChMA increases storage modulus 8-12-fold, reduces pore size, and confers antibacterial activity against gram-positive and gram-negative bacteria, addressing infection susceptibility. The fortified HMW hybrid (HMWH) network enables sustained, localized NMase delivery that outperforms GelMA alone in resisting degradation and controlling release kinetics. NMase-loaded HMWH (N-HMWH) gels enhance DDF proliferation and migration in vitro, correlating with reduced focal adhesion size and increased turnover. In a diabetic rat model, N-HMWH patches achieve superior wound closure, outperforming EGF therapy, with robust epidermal regeneration, neovascularization, and collagen deposition. This work establishes glycocalyx-targeting hydrogels as a new class of wound therapeutics addressing the root cause of diabetic fibroblast failure, not just compensating with growth factors.

bioengineering↗

bioRxiv2026

BK Channels Orchestrate Cardiac Homeostasis Through Mitochondrial Uncoupling Proteins

BK channels, coded by the Kcnma1 gene, integrate voltage and intracellular Ca2+ signals and are recognized for their roles in smooth muscle and neuronal excitability. However, their contribution to baseline cardiac physiology remains poorly defined. Here we uncover a fundamental function for BK channels in maintaining normal cardiac performance, independent of pathological stress. Using non-invasive echocardiography, transcriptional profiling, and mechanistic analyses, we demonstrate that Kcnma1 deletion disrupts ventricular function, and remodels metabolic and stress-response pathways. Transcriptomic profiling revealed selective downregulation of mitochondrial uncoupling proteins (UCPs) and suppression of the PGC-1/FOXO3a axis, without broad loss of oxidative phosphorylation components. Enhancing UCP expression restored cardiac performance, indicating that mitochondrial uncoupling and redox control constitute key downstream effectors of BK signaling. Together, these results identify a physiological role for BK channels in maintaining myocardial function and define a mitochondrial BK-UCP axis, critical for cardiac homeostasis.

cell biology↗

bioRxiv2026

A Scalable High-Density Microwell Assay for Single-Cell Clonal Expansion Profiling

Traditional clonogenic assays remain central to evaluating the self-renewal capacity of tumor cells. However, the assay relies on subjective endpoint measurements, is restricted to two-dimensional monolayer growth, and lacks the single cell resolution required to resolve heterogeneous expansion behaviors. We describe a high-density microwell array-based platform for quantitative assessment of single cell clonogenic growth outcomes, defined by cell count distributions spanning non-dividing, slow-dividing, and fast-dividing three-dimensional colony forming phenotypes. This approach links initial single-cell occupancy to defined growth outcomes across thousands of indexed microwells per well. The platform integrates high-density, low-adhesion microwell arrays within industry standard device plate formats and an automated image analysis pipeline incorporating machine learning, enabling parallel quantification of spatially indexed founder-derived microwells using widely accessible automated imaging systems. The assay was implemented in both 4-well and 96-well plate formats to evaluate reproducibility and scalability across different plate configurations. Using three glioblastoma cell lines as model systems, we demonstrate reproducible single founder occupancy and consistent clonal growth outcome distributions across replicate formats. This integrated microscale assay platform enables systematic quantitative characterization of clonogenic expansion capacity at single cell resolution and is compatible with applications in cancer biology, therapeutic testing, and functional single cell phenotyping. By resolving single-cell persistence, limited expansion and high expansion outcomes within a scalable high-density format, this approach expands the analytical resolution of single cell clonogenic profiling beyond traditional binary colony scoring.

cell biology↗

bioRxiv2025

Filling knowledge gaps on vulnerable electric ray Narcine timlei (Elasmobranchii: Torpediniformes: Narcinidae) through ecological and biological studies

Narcine timlei (Bloch & Schneider, 1801), commonly known as the brown numbfish, recently moved from Data Deficient to Vulnerable in IUCN Red List. However, the biological information on the species is still lacking. In the present study, we provide an integrative assessment of populational dynamics on N. timlei. Morphological and molecular analyses ascertain the species identity as N. timlei. A total of 331 individuals were examined, comprising 156 males and 175 females, including 13 gravid, bringing an overall sex ratio of 1:1.21. We observed the highest specimens during the post-monsoon season (33.5%), and minimum during summer (18%). Length-weight relationships indicated negative allometric growth (b=2.78). Most specimens fall under the 17-20 cm size class, and a few exceeded 23 cm. Pregnant females carried 2 to 9 embryos. A total of 69 embryos from 13 females were obtained. Embryo length and weight were 5.18 {+/-} 0.22 cm and 2.02 {+/-} 0.36 g, respectively. No correlation was observed between maternal length and fecundity (R2 = 0.0036). Male to female embryo ratio was skewed to 1:1.3. The average length of electric organs in adults was 4.30 {+/-} 0.47 cm. Mean electrocyte count for adults was 117.55 {+/-} 14.48, while in embryos it was 123.36 {+/-} 15.9, indicating early developmental establishment of electrocyte structure in N. timlei. We strongly recommend long term species-specific monitoring and management measures to ensure conservation of this ecologically significant but often overlooked batoid.

zoology↗

bioRxiv2025

PLGA Soya Lecithin Based Hybrid Nanocomposite for Targeted Topical Delivery of Resveratrol in Psoriasis Management

Psoriasis is a recurring autoimmune-driven inflammatory skin disorder characterised by red, scaly, dry skin patches, keratinocyte hyperproliferation and infiltration of inflammatory cells, impacting millions globally. Patients with psoriasis often require repeated treatments with corticosteroids, which are often associated with significant side effects and comorbidities. Therefore, there is a growing need for advanced nanomedicine-based therapeutic approaches. Resveratrol (RSV), a plant-derived polyphenol, has shown promising anti-inflammatory and antioxidant properties for the treatment and management of psoriasis. However, its therapeutic potential is limited due to its poor solubility, low stability, and reduced skin penetration. To overcome these challenges, we have developed dual-carrier conjugated nanoparticles (RSVNPL) composed of soya lecithin and poly(lactic-co-glycolic acid) (PLGA). To further enhance its ease of application and prolong skin residence time, RSVNPL was incorporated into a carbomer 974P-based hydrogel system, forming RSVNPGel. Our formulation demonstrated improved physicochemical properties, including enhanced spreadability, superior skin adhesiveness and stability. Ex vivo skin permeation studies confirmed a significantly higher accumulation of RSV in the epidermis and dermis compared to free drugs. In vivo, efficacy was investigated using imiquimod (IMQ)-induced psoriatic mouse model, where RSVNPGel treatment led to a notable reduction in epidermal thickness, inflammatory cell infiltration, and key histopathological features associated with psoriasis. Additionally, serum biochemical analysis and organ histology established the biocompatibility and safety of the formulation. Overall, RSVNPGel presents a promising nanocarrier-based strategy for the effective topical management of psoriasis, enhancing RSV delivery and therapeutic efficacy while minimizing systemic exposure. Research HighlightsO_LIHybrid nanoparticles composite of soya lecithin and PLGA were developed for RSV delivery. C_LIO_LIThe formulation enhanced skin penetration, retention and created a better skin adhesive profile in psoriatic lesions. C_LIO_LISignificant anti-psoriatic effects were achieved in a preclinical model. C_LIO_LIThe system showed excellent safety and biocompatibility. C_LI Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=101 SRC="FIGDIR/small/648585v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@e28c4corg.highwire.dtl.DTLVardef@1690c80org.highwire.dtl.DTLVardef@1b3e4bforg.highwire.dtl.DTLVardef@16ec0ad_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

bioRxiv2024

Inclusion of Hsp70 co-chaperone and RNA binding activities facilitates optimal function of spliceosomal disassembly factor Cwf23 in intron-rich Schizosaccharomyces pombe

Cwc23 is an essential J-domain protein that collaborates with the disassembly factor Ntr1 to facilitate spliceosomal disassembly. Although Cwc23 orthologs have been identified in spliceosomal extracts of many eukaryotes, their functionality in intron-rich eukaryotes remains largely unexplored. Here, we investigate the functionality of Cwf23, an ortholog of Cwc23 in Schizosaccharomyces pombe. Our study reveals that while the interaction between Cwf23 and SpNtr1 is conserved, it is not essential in S. pombe. Additionally, the RNA recognition motif (RRM) in Cwf23 is crucial for its function, as mutations in the RRM affect both growth and pre-mRNA splicing. Unlike its budding yeast counterpart, the J-domain of Cwf23 is indispensable, as J-domain mutants cannot support cell viability. These findings suggest a new paradigm in which the presence of a functional RRM and the essential nature of the J-domain underscore the increased requirements for an RNA-binding Hsp70 co-chaperone machinery in spliceosomal remodelling in "intron-rich" eukaryotes.

molecular biology↗

bioRxiv2023

One Health Assessment of an Urban Temporary Settlement Reveals Gut Microbiome Serving as Antimicrobial Resistance Gene Reservoir

Antimicrobial resistance (AMR) is an emerging and growing global health challenge that could result in 10.2 million deaths annually by 2050. The unrestricted and haphazard use of antibiotics is contributing to the rapid emergence and spread of AMR, and the problem is exacerbated by release of untreated waste water from high-risk sources like hospitals into rivers. Bacteria often develop resistance through horizontal gene transfer mechanism and gut flora can act as a source for new Antimicrobial Resistance Genes (ARG). Upcoming methods like metagenomics can identify the resistance profile (AMR) of gut microbiome, and detect bacterial infections that otherwise go unnoticed. Our study focused on understanding the presence of AMR mutations and gene transfer dynamics in human, animal and environmental samples collected in one of the temporary settlements of Kathmandu (Nepal) using One Health approach. Current AMR reporting based on clinical cases is limited and does not provide information on specific pathogen and associated AMR genes-our study is an effort to contribute information to fulfill this gap. Twenty-one samples were collected from a temporary settlement in Thapathali (Kathmandu), which included fecal samples from birds (n=3) and humans (n=14), and environmental samples (n=4). Microbiological assessment was carried out based on 16S sequence metagenomic analysis using MiSeq (Illumina, USA). Taxonomic classification on obtained 16S sequences were determined by using Metaphlan 2 and Qiime 2 bioinformatics tools. ShortBRED was used to classify ARG and virulence factors, and WAFFLE was used for horizontal gene transfer event prediction. The network analysis was carried out using Gephi v0.9 and the ResistoXplorer web tool to identify ARG in the collected samples. Prevotella spp. was the dominant gut microbiome in humans. We detected diverse phages and viruses, including Stx-2 converting phages. 72 virulence factors and 53 ARG subtypes were detected, with poultry samples having the highest number of subtypes. The cluster and network analysis showed a strong association between gut microbiome and ARG, which was also supported by Horizontal Gene Transfer (HGT) analysis. One-Health interface showed ARG dynamics and revealed gut microbiomes of humans and animals serving as a reservoir for the circulating ARG.

genomics↗