Biology subjects

Barman, M.

Publications and source records attributed to Barman, M..

7 recordsLinked to original sources

bioRxiv2026

Platelet-derived CXCL7 induces neutrophil extracellular traps via CXCL7/CXCR2 axis, exacerbating the pathogenesis of diabetic retinopathy

BackgroundProlonged hyperglycemia in diabetes activates platelets and immune cells, forming platelet-immune complexes that damage blood vessels in the retina. However, the role of platelet-neutrophil interactions and neutrophil extracellular traps(NETs) in the development of diabetic retinopathy (DR) was not well studied. In this study, we investigated the mechanisms underlying platelet-mediated NET formation in DR. MethodologyPlatelet activation markers, platelet-neutrophil aggregates (PNA), and NETs markers were assessed by flowcytometry, and the circulatory level of inflammatory markers was measured by Luminex assays in healthy control(HC), type 2 diabetes mellitus(T2DM), non-proliferative DR(NPDR) and proliferative DR(PDR) subjects. In vitro studies investigated platelet-neutrophil interaction in NETs formation using an immunofluorescence assay. Proteomics analysis identified the mechanistic regulators of platelet-induced NETs in DR. Platelet pellet and plasma CXCL7 were quantified using western blot and ELISA, respectively. The role of the CXCL7/CXCR2 axis in inducing NETs formation was examined using CXCL7 recombinant protein, anti-CXCL7 antibody and CXCR2 antagonist (SB225002). ResultsPlatelet activation markers (p-selectin & PF4), PNA, and NETs markers (%NETs, proteinase-3 (PR3), neutrophil elastase (NE)) were significantly increased in the DR group. In vitro studies confirmed that DR-platelets aggregate with healthy neutrophils and form NETs compared to T2DM and HC-platelets. Furthermore, platelet activation and NETs markers were positively correlated with pro-angiogenic (ANGPT2, VEGFA) and inflammatory markers (IL18, ICAM1). In vitro studies reveal that NETs induce inflammation, endothelial dysfunction, disrupt the endothelial monolayer and exacerbate angiogenesis in RF/6A endothelial cell spheroids. Proteomics analysis of platelet-induced NETs in DR revealed dysregulation of proteins involved in platelet activation and NET formation, including CXCL7. Furthermore, increased CXCL-7 levels were observed in platelet pellet and plasma samples from the DR group. Additionally, CXCL7-treated neutrophils formed NETs via the CXCR2 receptor, and inhibition of NETosis was observed in neutrophils exposed to an anti-CXCL-7 antibody and a CXCR2 antagonist. ConclusionOur findings revealed that platelets released CXCL-7 induce NETs formation via the CXCL7/CXCR2 axis and blockade of CXCL7/CXCR2 axis inhibits the NETosis in DR, thereby inhibiting the pathogenesis of DR. Circulating CXCL7 serves as a potential prognostic marker, and the CXCL7/CXCR2 axis may be a therapeutic target for the treatment of DR. What Are the Clinical Implications?Platelets have emerged as immune cells, and platelet-neutrophil interactions are reported to play a significant role in the pathogenesis of various metabolic diseases. The role of platelet-neutrophil interactions and neutrophil extracellular traps (NETs) in the development of diabetic retinopathy (DR) remains poorly understood. Investigating the mechanistic regulators of platelet-mediated NETs in DR is crucial for identifying new therapeutic approaches. Our study observed increased platelet activation, platelet-neutrophil aggregates and NETs among DR subjects. Further, DR-platelet induces NET in healthy neutrophils, and NETs induce inflammation, angiogenesis, and disrupt endothelial barrier function in RF/6A cells in vitro. These findings strengthen the evidence that platelet-neutrophil interactions play a major role in DR pathogenesis. Proteomic analysis identified CXCL7 as a mechanistic regulator of platelet-induced NETs formation in DR. Inhibitors that target the platelet-derived CXCL7/CXCR2 axis for NETosis can be used for the prevention of retinal injury in DR. Overall, our work emphasises the mechanistic understanding of platelet-neutrophil interactions and CXCL7/CXCR2 axis as a therapeutic target for inhibiting the pathogenesis of DR. Graphical abstractScheme for the platelet-derived CXCL7 regulation of NETs in DR. Activated platelets release CXCL7 and platelet aggregates with neutrophils to form NETs via the CXCL7:CXCR2 axis. Blockade of the CXCL7:CXCR2 axis by anti-CXCL7 antibody and CXCR2 inhibitor prevents NETs formation in DR.

immunology↗

bioRxiv2025

Synergistic 3D, multispectral, and thermal image analysis via supervised machine learning for improved detection of root rot symptoms in hydroponically-grown flat-leaf parsley

Root rot in hydroponically-grown leafy vegetables is difficult to detect via conventional manual and machine vision-based approaches as symptoms of infection are not clearly visible on the canopy at earlier stages of infection. Hence, the present study investigates the potential of using machine learning for assessing canopy information obtained from multiple imaging platforms synergistically to improve root rot detection. Herein, flat-leaf parsley seedlings were grown in an experimental hydroponic vertical farm and inoculated with Pythium irregulare and Phytophthora nicotianae. Subsequently, the seedlings were imaged via 3D, multispectral, and thermal sensors at various stages of growth to obtain twenty-six image-based plant features. Following a preliminary screening of redundant features via regression analysis, data for seventeen image features associated with morphometric, spectral, and thermal attributes was co-analyzed using supervised machine learning by Support Vector Machines (SVM). Exhaustive feature selection using different SVM kernels and maximum feature thresholds was performed to identify optimal feature subsets. It was observed that combining parameters obtained from all three imaging platforms enabled better identification of infected samples (>99%) than using a higher number of attributes from individual imaging systems. In addition, model performance was improved considerably by including temporal information during model training. Hence, it may be inferred that fusion of data from multiple imaging systems and using it with temporal information can enable better real-time high-throughput monitoring of root rot.

plant biology↗

bioRxiv2024

3-epicaryoptin induces G2/M phase cell cycle arrest and apoptosis in human breast cancer cells by disrupting the microtubule network, an in vitro and in silico investigation

Breast cancer (BC) is a prevalent form of cancer observed in women across the globe, constituting over a quarter of all female BC cases. The treatment of BC continues to require significant efficacy, aiming to achieve high success rates while minimizing adverse effects on the body as a whole. In the current study, 3-epicaryoptin was tested for the molecular mechanism of its anti-cancer activity in the human breast cancer cell line, MCF-7. We investigated cell viability by MTT assay, cell cycle kinetics and apoptosis, immunofluorescence straining, molecular modelling, and ADMET profiling. MTT assay results showed that 3-epicaryoptin was found cytotoxic against MCF-7 cells with an IC50 value of 344.64 {micro}g mL-1 for 48 h. Flow cytometric analysis exhibited that 3-epicaryoptin halted the MCF-7 cells in the G2/M phase and subsequently induced apoptosis in a time-dependent manner. Our immunofluorescence studies indicated that 3-epicaryoptin inhibited microtubule polymerization in MCF-7 cells. Furthermore, molecular docking followed by molecular dynamics (MD) simulation studies demonstrated the ability of 3-epicaryoptin to interact with the tubulin protein at the colchicine binding pockets. Overall, our results suggest that 3-epicaryoptin can inhibit the proliferation of human breast cancer cells by depolymerizing of cellular microtubule networks, which causes cell cycle arrest and promotes apoptotic cell death. Therefore, it has been indicated that the natural product 3-epicaryoptin exhibited considerable promise as a potent therapeutic agent capable of inducing apoptosis in breast cancer cells.

cancer biology↗

bioRxiv2021

Cytotoxic and micronuclei inducing effects of petroleum ether fraction of leaf aqueous extract of Clerodendrum viscosum Vent. in Allium cepa root tip cells

Clerodendrum viscosum is a traditionally used medicinal plant. The present study aimed to analyze a detailed cytotoxic effect of the nonpolar petroleum ether fraction (AQPEF) of leaf aqueous extract of C. viscosum Vent. (LAECV) in Allium cepa root tip cells. The LAECV was fractionated with petroleum ether and tested for A. cepa toxicity at early hours (2 and 4 h treatment) at concentrations 0, 50, 100, 150 and 200 {micro}g mL-1. The highest aberrant cell percentage (10.45{+/-}0.46%) was scored from 150 {micro}g mL-1 followed by100 {micro}g mL-1 (8.75{+/-}0.26%) concentration at 4 h treated samples. The AQPEF treatment induced a significant (p<0.0001) increase in micronuclei frequency; 4.31{+/-}0.33, 5.08{+/-}0.13, 5.05{+/-}0.22 and 3.05{+/-}0.37% respectively at concentrations 50, 100, 150, and 200 {micro}g mL-1. The highest polyploid frequency (20.14{+/-}0.68 %) induced with 100 {micro}g mL-1 of AQPEF at 16 h recovery. 150 {micro}g mL-1 is the most effective concentration of AQPEF to decipher its activity similar to colchicine (150 {micro}g mL-1). In summary, the present study indicates petroleum ether is suitable for extraction of the active phytochemicals of LAECV having cytotoxic effects on A. cepa root tip cells. The AQPEF has colchicine like micronuclei, polyploidy, and mitotic abnormality inducing potentials in A. cepa root apical meristem cells.

pharmacology and toxicology↗

bioRxiv2021

ProLung™-budesonide Inhibits SARS-CoV-2 Replication and Reduces Lung Inflammation

BackgroundInhaled budesonide benefits patients with COVID-19. ProLung-budesonide enables the sustained, low dose administration of budesonide within a delivery vehicle similar to lung surfactant. ProLung-budesonide may offer anti-inflammatory and protective effects to the lung in COVID-19, yet its effect on SARS-CoV-2 replication is unknown. ObjectiveTo determine the efficacy of ProLung-budesonide against SARS-CoV-2 infection in vitro, evaluate its ability to decrease inflammation, and airway hyperresponsiveness in an animal model of lung inflammation. MethodsSARS-CoV-2-infected Vero 76 cells were treated with ProLung-budesonide ([0.03- 100 g/ml]) for 3 days, and virus yield in the supernatant was measured. Ovalbumin-sensitized C57BL/6 mice received aerosolized (a) ProLung-budesonide weekly, (b) only budesonide, either daily or weekly, or (c) weekly empty ProLung-carrier (without budesonide). All treatment groups were compared to sensitized untreated, or normal mice using histopathologic examination, electron microscopy (EM), airway hyperresponsiveness (AHR) to Methacholine (Mch) challenge, and eosinophil peroxidase activity (EPO) measurements in bronchioalveolar lavage (BAL). ResultsProLung-budesonide showed significant inhibition on viral replication of SARS-CoV-2-infected cells with the selectivity index (SI) value > 24. Weekly ProLung-budesonide and daily budesonide therapy significantly decreased lung inflammation and EPO in BAL. ProLung-budesonide localized in type II pneumocytes, and was the only group to significantly decrease AHR, and EPO in BAL with Mch challenge ConclusionsProLung-budesonide significantly inhibited viral replication in SARS-CoV-2 infected cells. It localized into type II pneumocytes, decreased lung inflammation, AHR and EPO activity with Mch challenge. This novel drug formulation may offer a potential inhalational treatment for COVID-19.

pharmacology and toxicology↗

bioRxiv2021

Appraisal of Cell Cycle Delay and Cytotoxicity Inducing Potential of 3-epicaryoptin in Allium Cepa L.

Diterpenoid 3-epicaryoptin (C26H36O9) is abundant in the leaves of Clerodendrum inerme, a traditionally used medicinal plant, having insect antifeedant activities. Here, we aim to explore the cytogenotoxic effects of compound 3-epicaryoptin in Allium cepa root apical meristem cells. A. cepa roots were treated with 3-epicaryoptin (100, 150 & 200 g mL-1 concentration) and the standard compound colchicine (200 g mL-1 concentration) for 2, 4, 4+16 h (4 h treatment followed by 16 h recovery). Cytogenotoxicity was analysed by studying the root growth retardation (RGR), mitotic index (MI), and cellular aberrations. The result showed statistically significant (p<0.01), concentration-dependent RGR effects of 3-epicaryoptin treatment compared with the negative control. Study of cell frequency in different phases of cell division observed a significant (p<0.001) increase in the metaphase cell percentage (66.2{+/-}0.58 %, 150 g mL-1) and which subsequently caused an increase in the frequency of MI (12.29{+/-}0.34 %, 150 g mL-1) at 4h of 3-epicaryoptin treatment and that was comparable with the colchicine action. The cytological study revealed that the 3-epicaryoptin treatment could induce different types of chromosomal abnormalities such as colchicine like metaphase, vagrant chromosomes, sticky chromosomes, anaphase-bridge, and an increased frequency of micronuclei and polyploid cells. These findings indicate that 3-epicaryoptin is cytogenotoxic, and thus C. inerme should be used with caution in traditional medicine.

pharmacology and toxicology↗

bioRxiv2020

Pro-metaphase arrest, polyploidy, micronuclei, and mitotic abnormality inducing agents isolation from leaf aqueous extract of Clerodendrum viscosum Vent.

Clerodendrum viscosum is a traditionally used medicinal plant. The present study aimed to analyze cell cycle delay, pro-metaphase arrest, and c-metaphase inducing effects of the petroleum ether fraction (AQPEF) of leaf aqueous extract of C. Viscosum (LAECV). The LAECV was fractionated with petroleum ether and its metaphase arrest, cell cycle delay, and c-metaphase inducing activities were tested on A. cepa root tip cells. The AQPEF induced cell cycle delay, and colchicine like metaphase, c-metaphase, in A. cepa root tip cells. Thus, the present study explores AQPEF as an active fraction of LAECV having metaphase arresting, cell cycle delay, and c-metaphase inducing potentials.

pharmacology and toxicology↗