Biology subjects

Ullah, N.

Publications and source records attributed to Ullah, N..

12 recordsLinked to original sources

bioRxiv2026

NheABC is a pH-dependent cytotoxin that contributes to the virulence of Bacillus cereus

Bacillus cereus is a Gram-positive bacterium widely distributed in food, soil, and plants. It is a spore-forming, facultative anaerobic bacterium known as a food-borne opportunistic human pathogen responsible for causing gastrointestinal and non-gastrointestinal infections, including wound-associated infections. In the present study, we report that among single-component, bi-component, and tripartite alpha pore-forming toxin (-PFT) producing bacteria, the tripartite NheABC toxin produced by B. cereus induced the maximum cell death in infected epithelial cells. Similar to its effects in 2D monolayers, NheABC exhibited potent cell toxicity in 3D spheroids and intestinal organoids, targeting their cell membrane and mitochondria. Moreover, using erythrocytes as a model system, we found that the cytolytic activity of NheABC is pH-dependent, and was markedly reduced at acidic pH (5.5). The pH-dependent biological activity of NheABC was further confirmed by a liposome leakage assay. Importantly, NheABC enhanced the colonization of B. cereus in a non-gastrointestinal murine wound infection model. Overall, our study highlights the role of pH in regulating NheABC-mediated cytotoxicity in mammalian cells, which may lead to the development of novel therapeutic strategies for managing B. cereus gastrointestinal and non-gastrointestinal infections.

microbiology↗

bioRxiv2025

Mechanisms involved in the lipid membrane tubulation activity of the pH-responsive MakA protein from Vibrio cholerae

Bacterial pore-forming toxins (PFTs) are key virulence factors that disrupt host cell membranes. The Vibrio cholerae cytotoxin MakA, a member of the ClyA structural family of -PFTs, exhibits a unique, pH-dependent ability to induce lipid membrane tubulation. This study elucidates the molecular mechanisms underlying this process. We identify the N-terminal histidine residue (His30) of MakA as a critical pH sensor that is indispensable for membrane tubulation but not required for membrane binding and cell lysis. Substitution of His30 with lysine, MakAH30K , abrogated tubulation without affecting membrane insertion, instead triggering aberrant membrane fusion events. Circular dichroism spectroscopy analysis revealed that the MakAH30K substitution attenuates the robust -helical transition observed in wild-type MakA upon lipid membrane binding, providing a molecular explanation for its functional defect. Furthermore, we established that membrane cholesterol contributes to MakA-induced tubulation of lipid membranes, as its absence or mutation of a putative cholesterol-binding motif MakAI236D&I237D abolishes both membrane association and tubule formation. A key implication of our work is the mechanistic uncoupling of membrane tubulation from cytolysis, revealing that MakA can perturb membranes via distinct pathways. These findings define a sophisticated activation mechanism where His30 protonation in the acidic host environment and specific cholesterol binding synergistically drive the conformational change necessary for helical oligomerization and membrane tubulation. We discuss our findings from the viewpoint of possible convergent evolution, where bacterial PFTs like MakA partly may mimic ancient membrane remodelers involved in membrane repair pathways, such as ESCRT-III, but for bacterial fitness in host interactions and pathogenesis. Moreover, the present study provides a foundation for possible development of therapeutic strategies aimed at inhibiting these critical interactions.

biochemistry↗

bioRxiv2025

Pathogenic strains of a gut commensal drive systemic platelet activation and thromboinflammation in lupus nephritis

Imbalances in the gut microbiome have been linked to increased intestinal permeability and disease flares in systemic lupus erythematosus (SLE). Our study revealed that patients with flares of lupus nephritis (LN) and intestinal expansions of the anaerobic commensal, Ruminococcus gnavus (RG), displayed whole blood transcriptome profiles indicative of platelet, neutrophil, and myeloid cell activation, a profile reminiscent of sepsis. Serum analysis confirmed elevated serum levels of Platelet Factor 4 and neutrophil extracellular traps, which significantly correlated with levels of IgG-antibody to a novel lipoglycan (LG) produced by pathogenic RG strains, which was also documented in an independent LN cohort. To test for causality, in vivo mouse models further demonstrated that gut colonization with LG-producing RG strains, as well as a single intraperitoneal challenge with an LG preparation, caused platelet activation and megakaryocytosis in bone marrow and spleen. Mice colonized with RG strains that produce LG developed cellular infiltration of the kidneys by neutrophils and monocytes. Hence, RG expansions during renal flares may identify a specific LN flare endotype driven by thromboinflammatory mechanisms. Antibodies that arise from immune exposure to the RG lipoglycan may serve as a surrogate biomarker, helping to elucidate the impact of the relationship between gut microbiota communities and clinical outcomes in patients afflicted by LN. [208] Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=164 SRC="FIGDIR/small/641288v1_ufig1.gif" ALT="Figure 1"> View larger version (74K): org.highwire.dtl.DTLVardef@1239f8eorg.highwire.dtl.DTLVardef@1c062caorg.highwire.dtl.DTLVardef@195ece2org.highwire.dtl.DTLVardef@1f3123f_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

bioRxiv2023

Indigenous probiotics Lactobacillus reuteri and Enterococcus faecium exhibit positive growth performance and disease prevention against extended-spectrum cephalosporin and fluoroquinolones resistant Salmonella enterica in broiler chicks.

ABSTRACTThe rapid increase in antibiotic resistance poses a global threat to public health, necessitating the development of effective antimicrobial alternatives. This study compared an indigenous probiotic mix containing Lactobacillus reuteri and two strains of Enterococcus faecium to a commercial probiotic blend ProtexinR on the growth performance, mortality rate, histomorphology, serum immunoglobulins, and intestinal microflora of broiler chickens challenged with two multi drug resistant Salmonella serovars, Typhimurium and Enteritidis. Two hundred and forty day-old broiler chicks were randomly assigned to six treatment groups for 4 weeks: the treatment groups were; birds continuously supplemented with only indigenous probiotic strains (108 CFU/mL) (IPRO-); birds challenged with Salmonella serovars 106 (CFU/mL) (PC+); birds continuously supplemented with indigenous probiotic strains and challenged with Salmonella serovars (IPRO+); birds supplemented with ProtexinR and challenged with Salmonella serovars (CM+); birds supplemented with only ProtexinR (CM-); and birds with no Salmonella challenge or probiotics (negative control; PC-). The results revealed that IPRO- diets significantly improved feed conversion ratio (FCR) and increased body weight (BW) (P [≤] 0.05). No effect of probiotic treatments was observed on IPRO- and CM- on relative organ weights as compared to the negative control (PC-). The Salmonella- challenged group PC+ had the highest (20%) mortality rate and lowest BW. The IPRO- had significantly lower FCR (1.55) compared to PC- (1.86) and PC+ (1.95). The broilers in the IPRO- group showed significantly increased serum concentrations of IgA and IgG relative to both control groups (P [≤] 0.05). Morphological analysis of the ileum revealed significant increases (P [≤] 0.05) in the villus height and villus height/crypt depth in birds fed IPRO- compared with the PC+. Cecal Lactobacillus and Enterococcus counts were the highest (P [≤] 0.05) and Salmonella counts were the lowest (P [≤] 0.05) in the IPRO- group compared to the Salmonella infected group PC+. These results indicated that indigenous probiotic strains Lactobacillus reuteri and Enterococcus faecium can be an effective and low-cost alternative compared to commercial probiotics in the Pakistan poultry industry.

microbiology↗

bioRxiv2023

Pre- and post-flowering impacts of natural heat events on yield components in wheat

O_LICONTEXT Wheat crops are highly sensitive to elevated temperatures and experience significant yield losses when short periods of heat occur at sensitive developmental phases. C_LIO_LIOBJECTIVE This research aimed at quantifying wheat responses of grain yield and yield components to heat indicators in fluctuating field conditions. C_LIO_LIMETHODS The impacts of high temperature on yield and its components were assessed for 20-35 wheat lines in irrigated multi-environment trials over three years. Genotypes were cultivated using a novel photoperiod extension method adjacent to some conventional yield plots with different sowing dates. In the photoperiod-extension method, either single stems or plant quadrates were tagged at specific growth stages and hand-harvested at maturity, while conventional plots were mechanically harvested at maturity. The impact of heat was estimated for events occurring at different developmental stages and different temperature thresholds (26 to 35{circ}C). C_LIO_LIRESULTS The closest correlation between heat events and grain number was observed for a threshold temperature of 28{circ}C pre-flowering. For individual grain weight, the correlation was statiscally the closest for threshold temperatures above 32{degrees}C post-flowering. In the tested environments, grain number was most sensitive to heat between 300 and 200{circ}Cd before flowering. For each hot hour (T > 28{circ}C) during this period, wheat genotypes lost an extra 0.25 grains spike-1 or 90 grains m-2 on average. Post-flowering heat reduced individual grain weight by 0.26 mg (at spike harvest) and grain yield by 2.44 g m-2 (at plot harvest) for every hot hour (T > 32{circ}C) between 0 and 500{circ}Cd after flowering. Heat impacts were the clearest for plants with synchronised phenology. In addition, results suggest that heat impacts can be quantified more reliably using finer time units (i.e. hot hours rather than days). C_LIO_LIONCLUSIONS Natural heat events strongly impacted grain number and individual grain weight of wheat for temperatures above 28-30{degrees}C in well-watered conditions. Reduction in grain number and individual grain weight were strongly associated with accumulated hot hours that occurred during 200-300{degrees}Cd before and 0-500{degrees}Cd after flowering, respectively. C_LIO_LIIMPLICATIONS The findings from this study will assist improvement for crop modelling in response to heat, development of relevant phenotyping methods and selection of cultivars with better adaptation to warmer environments. C_LI

plant biology↗

bioRxiv2022

Integrated cyto-physiological and proteomic analyses reveal new insight into CMS mechanism in a novel upland cotton CMS line LD6A

Cytoplasmic male sterile (CMS) system has extensively been exploited for hybrid vigor in plant breeding programs. However, its application in many crops is limited due to poor understanding of molecular mechanism of fertility restoration. Using advanced analytical approaches, we elucidated molecular pathways regulating CMS induction and fertility restoration in cotton. Reproductive structures of a novel CMS (LD6A) and its maintainer (LD6B) line were analyzed for physiological and proteomic changes during the development process. Significant differential expression of proteins, such as Abrin, malate dehydrogenase, malic enzyme, isocitrate dehydrogenase, histone acetyltransferase was observed in CMS and its maintainer line. Transmission electron micrographs of anther tapetum showed that inner ridge of CMS mitochondria was relatively indistinct than that of LD6B with narrower membranous space at tetrad stage. Further, relatively higher reactive oxygen species were accumulated in the anther of CMS than its maintainer line at pollen mother cell and tetrad stage. We suggest that abnormal sequence of mitochondrial ribosome gene rps4 and rpl10 and high expression of ribosome-inactivating protein gene Abrin in CMS line damaged mitochondrial membrane and consequently induced pollen sterility. These data provide new insight into CMS mechanism in cotton crops and a tool to develop new CMS germplasm resources.

developmental biology↗

bioRxiv2021

SARS-CoV-2 papain-like protease PLpro in complex with natural compounds reveal allosteric sites for antiviral drug design

SARS-CoV-2 papain-like protease (PLpro) covers multiple functions. Beside the cysteine-protease activity, PLpro has the additional and vital function of removing ubiquitin and ISG15 (Interferon-stimulated gene 15) from host-cell proteins to aid coronaviruses in evading the hosts innate immune responses. We established a high-throughput X-ray screening to identify inhibitors by elucidating the native PLpro structure refined to 1.42 [A] and performing co-crystallization utilizing a diverse library of selected natural compounds. We identified three phenolic compounds as potential inhibitors. Crystal structures of PLpro inhibitor complexes, obtained to resolutions between 1.7-1.9 [A], show that all three compounds bind at the ISG15/Ub-S2 allosteric binding site, preventing the essential ISG15-PLpro molecular interactions. All compounds demonstrate clear inhibition in a deISGylation assay, two exhibit distinct antiviral activity and one inhibited a cytopathic effect in a non-cytotoxic concentration range. These results highlight the druggability of the rarely explored ISG15/Ub-S2 PLpro allosteric binding site to identify new and effective antiviral compounds. Importantly, in the context of increasing PLpro mutations in the evolving new variants of SARS-CoV-2, the natural compounds we identified may also reinstate the antiviral immune response processes of the host that are down-regulated in COVID-19 infections.

biochemistry↗

bioRxiv2021

A robust field-based method to screen heat tolerance in wheat

Wheat is highly sensitive to elevated temperatures, particularly during pollen meiosis and early-to-mid grain filling. The impact of heat stress greatly depends on the plant developmental stage. Thus, germplasm ranking for heat tolerance in field trials may be confounded by variations in developmental phase between genotypes at the time of heat events. A photoperiod-extension method (PEM) was developed allowing screening of 35 diverse genotypes at matched developmental phase despite phenological variations. Paired trials were conducted to compare the new PEM against conventional field screening in plots. In the PEM, plants were sown in single rows or small plots. Artificial lighting was installed at one end of each row or plot to extend day length, inducing a gradient of flowering times with distance from the lights. Individual stems or plot quadrats of each genotype were tagged at flowering. Late-sown plants received more heat shocks during early to mid grain filling than earlier sowings, suffering reductions in both individual grain weight (IGW) and yield. IGW was reduced by 1.5 mg for each additional post-flowering day with temperature > 30{degrees}C. Significant genotypic differences in heat tolerance ranking were observed between PEM versus conventional plot screening. Strong correlations between trials experiencing similar degree of heat were found both for IGW and for total grain weight with the PEM either with individual-stem tagging (e.g. average r of 0.59 and 0.54, respectively for environments with moderate postflowering heat) or quadrat tagging (r of 0.53 and 0.47). However, correlations for IGW and yield in these environments were either poor or negative for conventional trials (e.g. average r of 0.11 and 0.12, respectively for environments with moderate postflowering heat). Accordingly, a PCA grouped genotypes consistently for heir performance across environments with similar heat stress in PEM trials but not in conventional trials. In this study, most consistent genotype ranking for heat tolerance was achieved with the PEM with tagging and harvesting individual spikes at matched developmental phase. The PEM with quadrat sampling provided slightly less consistent rankings but appears overall more suitable for high-throughput phenotyping. The method promises to improve the efficiency of heat tolerance field screening, particularly when comparing genotypes of different maturity types.

plant biology↗

bioRxiv2021

ASSOCIATION STUDY OF SIX CANDIDATE GENES WITH MAJOR DEPRESSIVE DISORDER IN THE NORTH-WESTERN POPULATION OF PAKISTAN

People around the world are currently affected by Major Depressive Disorder (MDD). Despite its many aspects, symptoms, manifestations and impacts, efforts have been made to identify the root causes of the disorder. In particular, genetic studies have concentrated on identifying candidate genes for MDD and exploring associations between these genes and some specific group of individuals. The aim of this research was to find out the association between single nucleotide polymorphisms in 6 candidate genes linked to the neurobiology of major depressive disorder in the North-Western population of Pakistan. We performed a case-control analysis, with 400 MDD and 232 controls. A trained psychiatrist or clinical psychologists evaluated the patients. Six polymorphisms were genotyped and tested for allele and genotype association with MDD. There were no statistical variations between MDD patients and healthy controls for genotypic and allelic distribution of all the polymorphisms observed. Thus, our analysis does not support the major role of these polymorphisms in contributing to MDD susceptibility, although it does not preclude minor impact. The statistically significant correlation between six polymorphisms and major depressive disorder in the studied population was not observed. There are inconsistencies in investigations around the world. Future research, including GWAS and association analysis on larger scale should be addressed for further validation and replication of the present findings.

genetics↗

bioRxiv2020

Massive X-ray screening reveals two allosteric drug binding sites of SARS-CoV-2 main protease

The coronavirus disease (COVID-19) caused by SARS-CoV-2 is creating tremendous health problems and economical challenges for mankind. To date, no effective drug is available to directly treat the disease and prevent virus spreading. In a search for a drug against COVID-19, we have performed a massive X-ray crystallographic screen of two repurposing drug libraries against the SARS-CoV-2 main protease (Mpro), which is essential for the virus replication and, thus, a potent drug target. In contrast to commonly applied X-ray fragment screening experiments with molecules of low complexity, our screen tested already approved drugs and drugs in clinical trials. From the three-dimensional protein structures, we identified 37 compounds binding to Mpro. In subsequent cell-based viral reduction assays, one peptidomimetic and five non-peptidic compounds showed antiviral activity at non-toxic concentrations. We identified two allosteric binding sites representing attractive targets for drug development against SARS-CoV-2.

biophysics↗

bioRxiv2020

Catalytic cleavage of HEAT and subsequent covalent binding of the tetralone moiety by the SARS-CoV-2 main protease

Here we present the crystal structure of SARS-CoV-2 main protease (Mpro) covalently bound to 2-methyl-1-tetralone. This complex was obtained by co-crystallization of Mpro with HEAT (2-(((4-hydroxyphenethyl)amino)methyl)-3,4-dihydronaphthalen-1(2H)-one) in the framework of a large X-ray crystallographic screening project of Mpro against a drug repurposing library, consisting of 5632 approved drugs or compounds in clinical phase trials. Further investigations showed that HEAT is cleaved by Mpro in an E1cB-like reaction mechanism into 2-methylene-1-tetralone and tyramine. The catalytic Cys145 subsequently binds covalently in a Michael addition to the methylene carbon atom of 2-methylene-1-tetralone. According to this postulated model HEAT is acting in a pro-drug-like fashion. It is metabolized by Mpro, followed by covalent binding of one metabolite to the active site. The structure of the covalent adduct elucidated in this study opens up a new path for developing non-peptidic inhibitors.

biophysics↗

bioRxiv2018

ANTI-MICROBIAL RESISTANCE IN AGENTS CAUSING URINARY TRACT INFECTIONS

BACKGROUNDUrinary Tract Infections are usually treated with empirical therapy by physicians based on previous knowledge of predictability of causative agents and their antimicrobial susceptibilities.\n\nOBJECTIVEThe objective of this study was to determine the frequency of various pathogens causing urinary tract infections and their antimicrobial susceptibility in patients presenting in out-patient department of a tertiary care hospital.\n\nMATERIALS AND METHODSThis descriptive cross sectional study was conducted in Out-Patient department of Urology of Benazir Bhutto Hospital during a period of 6 months from January 2017 to June 2017 after ethical approval from institutional research forum of Rawalpindi Medical University. 1000 patients (12 years old or above) that were clinically suspected for urinary tract infections were included in this study. Patients with co-morbidities like diabetes mellitus, renal pathologies, Immunodeficiency disorders, malignancies and congenital urogenital disorders were also excluded. Recipients of corticosteroid therapy or with a history of intake of broad spectrum antibiotics in previous 15 days were also excluded. Modified Kirby-Bauer disc diffusion method was used for determining the antimicrobial resistance against various antimicrobials.\n\nRESULTSA total of 530 (53%) isolates were found to be culture positive for E.coli(77.4%),Klebsiella (6.4%), Enterobacter (6.0%), Pseudomonas (3.8%), Staphylococcus saprophyticus (3.4%), Citrobacter (1.1%) and Morganella (0.4%).\n\nAntimicrobial resistance against commonly used antimicrobials was alarmingly high\n\nCONCLUSIONSurveillance of trends of antimicrobial susceptibility pattern is highly important.

microbiology↗