HypoxamiR-210-3p regulates mesenchymal stem cells proliferation via P53 & Akt.

Transplanted stem cells (˃95%) into ischemic myocardium die because of unfavourable conditions. Moreover, hypoxia role in the cell cycle regulation has been studied in transformed/immortalized cell lines which may have altered cell cycle regulators and/or mutated and, can't be transplanted in patients. We quest to find out the mechanism of cell cycle regulation in mesenchymal stem cells (MSC) to regulate its survival and proliferation in repair processes. Additionally, critically analysed role of hypoxamiR-210-3p, and cell cycle regulators that can regulate cell proliferation under hypoxic conditions. Bone marrow-derived MSC (BM-MSC) isolated from young male Fischer-344 rats by flushing the cavity of femur and propagated in vitro under 1% hypoxia for 72 h showed an increased in cell proliferation ([Formula: see text] 30%, p < 0.05) compared to normoxia. miR-210-3p, role in cell proliferation under hypoxic condition was confirmed by knockdown. Loss of function studies with transfection of anti-mir-210-3p, we observed decrease in proliferation of BM-MSC under hypoxia. Furthermore, BM-MSC proliferation due to miR-210-3p was confirmed using CFSE assay and flow cytometry, in which more cells were observed in S-phase. Mechanistically, western blot analysis showed miR-210-3p inhibition upregulates p53 and p21 expression and subsequent decrease in pAkt under hypoxia. On contrary, CFSE and Western blot under normoxic conditions showed downregulation of p53 and p21 whilst upregulation of pAkt indicated the key role of miR-210-3p in BM-MSC proliferation. Our results demonstrate the role of miR-210-3p in BM-MSC proliferation under both hypoxic and normoxic conditions and illustrate the potential mechanism via the regulation of pAkt, p53 and p21.

Engineering medicinal plant-derived CYPs: a promising strategy for production of high-valued secondary metabolites.

MAIN CONCLUSION: Cytochorme P450s (CYPs) play a critical role in the catalysis of secondary metabolite biosynthetic pathways. For their commercial use, various strategies for metabolic pathway engineering using CYP as a potential target have been explored. Plants produce a vast diversity of secondary metabolites which are being used to treat various ailments and diseases. Some of these metabolites are difficult to obtain in large quantities limiting their industrial use. Cytochrome P450 enzymes (CYPs) are important catalysts in the biosynthesis of highly valued secondary metabolites, and are found in all domains of life. With the development of high-throughput sequencing and high-resolution mass spectrometry, new biosynthetic pathways and associated CYPs are being identified. In this review, we present CYPs identified from medicinal plants as a potential game changer in the metabolic engineering of secondary metabolic pathways. We present the achievements made so far in enhancing the production of important bioactivities through pathway engineering, giving some popular examples. At last, current challenges and possible strategies to overcome the limitations associated with CYP engineering to enhance the biosynthesis of target secondary metabolites are also highlighted.

PIP-seq identifies novel heterogeneous lung innate lymphocyte population activation after combustion product exposure.

Innate lymphoid cells (ILCs) are a heterogeneous population that play diverse roles in airway inflammation after exposure to allergens and infections. However, how ILCs respond after exposure to environmental toxins is not well understood. Here we show a novel method for studying the heterogeneity of rare lung ILC populations by magnetic enrichment for lung ILCs followed by particle-templated instant partition sequencing (PIP-seq). Using this method, we were able to identify novel group 1 and group 2 ILC subsets that exist after exposure to both fungal allergen and burn pit-related constituents (BPC) that include dioxin, aromatic hydrocarbon, and particulate matter. Toxin exposure in combination with fungal allergen induced activation of specific ILC1/NK and ILC2 populations as well as promoted neutrophilic lung inflammation. Oxidative stress pathways and downregulation of specific ribosomal protein genes (Rpl41 and Rps19) implicated in anti-inflammatory responses were present after BPC exposure. Increased IFNγ expression and other pro-neutrophilic mediator transcripts were increased in BPC-stimulated lung innate lymphoid cells. Further, the addition of BPC induced Hspa8 (encodes HSC70) and aryl hydrocarbon transcription factor activity across multiple lung ILC subsets. Overall, using an airway disease model that develops after occupational and environmental exposures, we demonstrate an effective method to better understand heterogenous ILC subset activation.

PIP-Seq identifies novel heterogeneous lung innate lymphocyte population activation after combustion product exposure.

Innate lymphoid cells (ILCs) are a heterogeneous population that play diverse roles in airway inflammation after exposure to allergens and infections. However, how ILCs respond after exposure to environmental toxins is not well understood. Here we show a novel method for studying the heterogeneity of rare lung ILC populations by magnetic enrichment for lung ILCs followed by particle-templated instant partition sequencing (PIP-seq). Using this method, we were able to identify novel group 1 and group 2 ILC subsets that exist after exposure to both fungal allergen and burn pit-related constituents (BPC) that include dioxin, aromatic hydrocarbon, and particulate matter. Toxin exposure in combination with fungal allergen induced activation of specific ILC1/NK and ILC2 populations as well as promoted neutrophilic lung inflammation. Oxidative stress pathways and downregulation of specific ribosomal protein genes ( Rpl41 and Rps19 ) implicated in anti-inflammatory responses were present after BPC exposure. Increased IFNγ expression and other pro-neutrophilic mediator transcripts were increased in BPC-stimulated lung innate lymphoid cells. Further, the addition of BPC induced Hspa8 (encodes HSC70) and aryl hydrocarbon transcription factor activity across multiple lung ILC subsets. Overall, using an airway disease model that develops after occupational and environmental exposures, we demonstrate an effective method to better understand heterogenous ILC subset activation.