Biology subjects

Chattopadhyay, P.

Publications and source records attributed to Chattopadhyay, P..

6 recordsLinked to original sources

bioRxiv2024

Investigating the potential role of intimin/invasin in Aeromonas hydrophila virulence in Labeo rohita: a host-microbe interaction study

Aeromonas hydrophila is a primary bacterial pathogen affecting freshwater fish, including Labeo rohita (rohu), leading to significant losses in aquaculture. This study investigates the probable role of intimin/invasin, known virulence factors in several bacterial pathogens, in the pathogenesis of A. hydrophila in L. rohita. Using an in-silico approach, we explore the genetic and structural features of these proteins to predict their potential function in facilitating infection. We analysed the distribution of invasin and intimin across 53 A. hydrophila genomes and examined their physicochemical properties, including molecular weight and stability. Additionally, we evaluated the secondary structures to understand their functional roles in host-pathogen interactions. Using homology-based modelling, we generated 3D (tertiary) structures of invasin and intimin and selected the most suitable models for in silico docking experiments with all eight Rohu (Labeo rohita) {beta}-integrins-a crucial step in understanding their interactions with the fish (Rohu) host cells. Due to the unavailability of the crystal structure of Rohu {beta}-integrins, we performed homology-based modelling prior to the docking experiments. Our findings reveal that invasin and intimin are present in only 6 of the 53 A. hydrophila strains examined, primarily in highly virulent strains previously reported. Notably, invasin lacks disulfide bonds and beta turns. 3D modelling indicates a significant binding affinity of invasin with all human {beta}-integrins, suggesting a critical role in host-pathogen interactions.

microbiology↗

bioRxiv2022

Diversity of Secretion System Apparatus in Tomato Wilt Causing Ralstonia solanacearum Strains: a Comparative Analysis Using in-silico Approach

Ralstonia solanacearum (Rs) species is the leading cause of bacterial wilt disease in a wide range of host plants worldwide. In the present study, secretion system analysis of five tomato pathogenic Rs strains was carried out in-silico. This paper describes a new protocol to identify the secretion system components i.e. SSCs (T1SS-T6SS, Flg, T4P, and Tad-Tat). A total of 865 SSCs were identified using the new protocol. Contributions of SSCs into core-secretion system apparatus (i.e. SSA) were also studied. Synteny was discovered among the secretion system apparatus (SSA) where relative frequency of SSCs to core-SSA is high (>20%) which includes T1SS, T2SS, T5SS, T4P, and Tad-Tat, but excludes T3SS, T4SS, and Flg. To the best of our knowledge, this is the first report indicating that during the evolution of Rs, most of the secretion system apparatus (T1SS, T2SS, T5SS, T4P, and Tad-Tat) were highly conserved and came from a single ancestor, while T3SS and T6SS may have arrived later, probably from horizontal gene transfer.

microbiology↗

bioRxiv2021

Simultaneous assessment of eight phosphorylated STAT residues in T-cells by flow cytometry.

Signal transducer and activator of transcription (STAT) proteins are a family of transcription factors controlling functions in immune responses and other cell types. Given their importance, we developed a flow cytometry panel to assess eight phosphorylated STAT residues in human T-cells, including six tyrosine residues across six STAT proteins (STAT1, STAT2, STAT3, STAT4, STAT5a, STAT6) and additional serine residues on STAT1 and STAT3. We applied this protocol to test the in vitro induction of pSTATs in response to CD3/CD28 activation and a panel of recombinant cytokines. We also assessed the pSTAT expression profiles of naive CD4+ T-cells polarized to Th1, Th2, Th17 or iTregs. pSTAT1(S727), pSTAT2(Y689) and pSTAT3(S727) were constitutively expressed in most T-cells, even in the absence of stimulation. For pSTAT1(S727) and pSTAT3(S727), we observed two positive states, high and low. Conversely, expression of pSTAT1(Y701), pSTAT3(Y705), pSTAT4(Y693) and pSTAT6(Y641) were absent in resting T-cells and only expressed with CD3/CD28 activation or with specific cytokines. Variable frequencies of pSTAT5a(Y694) expression were observed in resting T-cells, which increased with activation or specific cytokine stimulation (e.g. IL-2). IFN{beta} stimulation enhanced frequencies of expressing cells for all pSTATs. Correlations among several pSTATs, particularly pSTAT1(S727)high and pSTAT3(S727)high were observed. While polarization resulted in increases in canonically associated pSTATs, other non-canonical pSTAT changes were also observed. Collectively, we developed, optimized, and tested a sensitive and rapid approach for simultaneously assessing phosphorylation of six STAT proteins. Using this approach, we made several novel observations of T-cell pSTAT induction in response to stimuli.

immunology↗

bioRxiv2021

Human fetal neural stem cell-derived astrocytes maintain glutamate transport after hypoxic injury in vitro

Astrocytes are the most abundant glial cells that play many critical roles in the central nervous system physiology including the uptake of excess glutamate from the synapse by Excitatory Amino Acid Transporters (EAATs). Among the EAATs, EAAT2 are predominantly functional, astrocyte-specific glutamate transporters in the forebrain. Hypoxic brain injury is a pathological phenomenon seen in various clinical conditions including stroke and neonatal hypoxic ischemic encephalopathy. Glutamate excitotoxicity is an important cause of neuronal cell death in disorders involving hypoxic brain injury. As findings from rodent models cannot always be reliably extrapolated to humans, we aimed to develop a homogenous population of primary human astrocytes to study the effect of hypoxic injury on astrocyte function, especially glutamate uptake. We successfully isolated, established and characterized cultures of human fetal neural stem cells (FNSCs) from aborted fetal brains. FNSCs were differentiated into astrocytes, and characterized by increased expression of the astrocyte markers, glial fibrillary acidic protein (GFAP), EAAT1 and EAAT2. A concomitant decrease in neural stem cell marker, Nestin, was observed. Differentiated astrocytes were exposed to various oxygen concentrations mimicking normoxia (20% and 6%), moderate and severe hypoxia (2% and 0.2% respectively). Interestingly, no change was observed in the expression of glutamate transporter, EAAT2 and glutamate uptake by astrocytes, even after exposure to hypoxia. Our novel model of human FNSC derived astrocytes exposed to hypoxic injury, establishes that astrocytes are able to maintain glutamate uptake even after exposure to severe hypoxia for 48 hours, and thus provides evidence for the neuroprotective role of astrocytes in hypoxic injury.

neuroscience↗

bioRxiv2021

Tonic interferon restricts pathogenic IL-17-driven inflammatory disease via balancing the microbiome

Maintenance of immune homeostasis involves a synergistic relationship between the host and the microbiome. Canonical interferon (IFN) signaling controls responses to acute microbial infection, through engagement of the STAT1 transcription factor. However, the contribution of tonic levels of IFN to immune homeostasis in absence of acute infection remains largely unexplored. We report that STAT1 KO mice spontaneously developed an inflammatory disease marked by myeloid hyperplasia and splenic accumulation of hematopoietic stem cells. Moreover, these animals developed inflammatory bowel disease. Profiling gut bacteria revealed a profound dysbiosis in absence of tonic IFN signaling, which triggered expansion of TH17 cells and loss of splenic Treg cells. Resolution of dysbiosis by antibiotic treatment averted the TH17 bias, and blocking IL17 signaling prevented myeloid expansion and splenic stem cell accumulation. Thus, tonic IFNs regulate gut microbial ecology, which is crucial for maintaining physiologic immune homeostasis and preventing inflammation.

immunology↗

bioRxiv2020

Potential malaria vector Anopheles minimus (species A) still persisting in North East India

BackgroundVector borne infectious diseases affect two third of the worlds human population and cause mortality in millions each year. Malaria remains one of the major killers in the Indian sub-continent and transmitted uninterruptedly by many efficient vectors and their sibling species. In North East India (NE), Anopheles minimus has been recognized as an important vector which shares majority of malaria cases. This study primarily focuses on to recognize the presence and distribution of sibling species of An. minimus in certain endemic area of NE India. MethodsAnopheles species were collected and identified using available morphological keys. The genomic DNA was extracted from the mosquito specimen and used to perform species specific PCR (ss PCR) for molecular identification of major malaria vector An. minimus sibling species ResultMorphological identification suggested the presence of An. minimus sl in low density in the study area. The specimen of An. minimus subjected to ss PCR confirmed the prevalence of only one sibling species namely, An. minimus A in Sialmari and Chandubi. ConclusionThough in low density, but malaria vector An. minimus is still present in certain endemic areas of NE India. The ss PCR assay employed presently suggested that An. minimus sibling species A is prevailing in the region. Presently used ss PCR assay was simpler, faster, cheaper and more readily interpreted than earlier assays. This information could be useful in understanding of current prevalence and distribution of An. minimus sibling species complex in NE region of India.

ecology↗