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Biology subjects

Saxena, R. K.

Publications and source records attributed to Saxena, R. K..

3 recordsLinked to original sources

InDels in an intronic region of gene Ccsmd04 coding for dormancy/auxin-associated protein controls sterility mosaic disease resistance in pigeonpea

Sterility mosaic disease presents a significant challenge to pigeonpea cultivation in the Indian subcontinent, potentially leading to total crop failure. The development of diagnostic molecular markers for SMD resistance can aid in improving SMD-resistant varieties. In this context, a QTL-seq approach identified genomic regions associated with SMD resistance using a recombinant inbred line generated from ICP8863 x ICPL87119. In total, 6,105 high-confidence variants were identified in the genomic region, namely, smdCc04 based on the delta SNP index. A genomic region smdCc04 on chromosome Cc04 spans 3.2 Mb (9.3 - 12.5 Mb) comprised of 6 missense variants and eight indels. A total of 211 candidate genes were identified from this region. 1 bp insertion, 21 bp insertion, 9 bp deletion, and 3 bp insertion at different intronic positions in 22 susceptible line leads to downregulation of Dormancy/auxin associated protein (Ccsmd04) resulting in loss of signaling in disease resistance pathways. The identified sites recognize four important disease and plant growth related transcription factors. A total of 4 Indels and 8 SNPs were validated from smdCc04 genomic regions using whole genome re-sequencing data and KASP genotyping on resistant and susceptible pigeonpea lines respectively. These markers will be used in pigeonpea breeding programs.

genomics↗

Poly-dispersed-acid-functionalized-single-walled carbon nanotubes(AF-SWCNTs) are potent inhibitor of BCG induced inflammatory response in macrophage cell lines

Present study is focussed on the modulation of Mycobacterium bovis BCG induced inflammatory response by poly-dispersed acid-functionalized single-walled carbon nanotubes (AF-SWCNTs) in macrophages. Flow cytometric and confocal microscopy studies indicated that both BCG and AF-SWCNTs were efficiently internalized by RAW 264.7 and MH-S macrophage cell lines and were essentially localized in the cytoplasmic area. The results indicated strong antioxidant activity of AF-SWCNTs in mitigating BCG induced oxidative and nitrosative stress. We also found a marked decline in expression of BCG induced pro-inflammatory genes like COX-2, iNOS, TNF-, IL-6 and IL-1{beta} on treatment with AF-SWCNTs at transcriptional level. Decline in expression of BCG induced COX-2 by AF-SWCNTs was also confirmed at protein level using Western blotting. Anti-inflammatory activity of AF-SWCNTs was further validated by our results showing that AF-SWCNTs treatment induced a precipitous decline in BCG induced release of Matrix Metalloproteinases MMP-2 and MMP-9 by macrophage cell lines, by using Gelatin zymography. Taken together, our results demonstrate potent anti-inflammatory role of AF-SWCNTs in alleviating BCG induced inflammation.

immunology↗

Modulation of cell adhesion and migration by poly-dispersed-acid-functionalized-single-walled carbon nanotubes in lung epithelial cells

Epithelial cell lining of the lung alveoli is under constant onslaught of airborne pathogens and pollutants that may cause injury and disruption of the epithelial lining. Repair mechanisms involve proliferation and migration of nearby healthy epithelial cells to the site of injury. Using murine LA4 and human A549 lung epithelial cell lines, and in vitro models of cell migration we have examined the modulation of cellular adhesion and migration by poly-dispersed acid-functionalized single-walled carbon nanotubes (AF-SWCNTs). Flow cytometric and confocal microscopy studies indicated that AF-SWCNTs were efficiently internalized by both cell lines and were localized essentially in the cytoplasmic area. In the scratch wound repair model, exposure to AF-SWCNTs blocked the filling of the scratched area of the cellular monolayers in both cells. Behaviour of the cells around the scratch area was examined in by live-cell imaging time-lapse micrography. The results indicated active cell proliferation around the scratch area that was totally blocked by AF-SWCNTs in LA4 cells and significantly inhibited in A549 cells. Cell migration across a porous membrane in transwell assay system also indicated a marked inhibition of migration of both cells across the membrane. Effect of AF-SWCNTs on the expression levels of important cell proteins involved in cell migration and adhesion were examined by western blotting and immunofluorescence staining. Expressions of proteins like {beta}-Catenin, NM-Myosin and Vimentin that play crucial role in cell migration were suppressed in AF-SWCNTs-exposed cells whereas the expression levels of E-cadherin and Claudin-1, involved in cell-cell adhesion remained unaltered. Our results provide an insight into the mechanism of repair of lung epithelial cell layers.

cell biology↗