The present study aims to identify the heat responsive genes and biological pathways in heat stressed buffalo mammary epithelial cells (MECs). The primary mammary epithelial cells of riverine buffalo were exposed to thermal stress at 42°C for one hour. The cells were subsequently allowed to recover at 37°C and harvested at different time intervals (30 min to 48 h) along with control samples (un-stressed). In order to assess the impact of heat stress in buffalo MECs, several in-vitro cellular parameters (lactate dehydrogenase activity, cell proliferation assay, cellular viability, cell death and apoptosis) and transcriptional studies were conducted. The heat stress resulted in overall decrease in cell viability and cell proliferation of MECs while induction of cellular apoptosis and necrosis. The transcriptomic profile of heat stressed MECs was generated using Agilent 44 K bovine oligonucleotide array and at cutoff criteria of ≥3-or ≤3 fold change, a total of 153 genes were observed to be upregulated while 8 genes were down regulated across all time points post heat stress. The genes that were specifically up-regulated or down-regulated were identified as heat responsive genes. The upregulated genes in heat stressed MECs belonged to heat shock family viz., HSPA6, HSPB8, DNAJB2, HSPA1A. Along with HSPs, genes like BOLA, MRPL55, PFKFB3, PSMC2, ENDODD1, ARID5A, and SENP3 were also upregulated. Microarray data revealed that the heat responsive genes belonged to different functional classes viz., chaperons; immune responsive; cell proliferation and metabolism related. Gene ontology analysis revealed enrichment of several biological processes like; cellular process, metabolic process, response to stimulus, biological regulation, immune system processes and signaling. The transcriptome analysis data was further validated by RT-qPCR studies. Several HSP (HSP40, HSP60, HSP70, HSP90, and HSPB1), apoptotic (Bax and Bcl2), immune (IL6, TNFα and NF-kβ) and oxidative stress (GPX1 and DUSP1) related genes showed differential expression profile at different time points post heat stress. The transcriptional data strongly indicated the induction of survival/apoptotic mechanism in heat stressed buffalo MECs. The overrepresented pathways across all time points were; electron transport chain, cytochrome P450, apoptosis, MAPK, FAS and stress induction of HSP regulation, delta Notch signaling, apoptosis modulation by HSP70, EGFR1 signaling, cytokines and inflammatory response, oxidative stress, TNF-alpha and NF- kB signaling pathway. The study thus identified several genes from different functional classes and biological pathways that could be termed as heat responsive in buffalo MEC. The responsiveness of buffalo MECs to heat stress in the present study clearly suggested its suitability as a model to understand the modulation of buffalo mammary gland expression signature in response to environmental heat load.
Austenite (γ-Fe, face centered cubic (FCC)) to ferrite (α-Fe, body centered cubic (BCC)) phase transformation in steel is of great significance from the point of view of industrial applications. In this work, using classical molecular dynamics simulations, we study the atomistic mechanisms involved during the growth of the ferrite phase embedded in an austenite phase. We find that the disconnections present at the inter-phase boundary assist in growth of the ferrite phase. Relatively small interface velocities (1.19 -4.67 m/s) confirm a phase change via massive transformation mechanism. Boundary mobilities obtained in a temperature range of 1000 to 1400 K show an Arrhenius behavior, with activation energies ranging from 30 -40 kJ/mol.
High strength and ductility, often mutually exclusive properties of a structural material, are also responsible for damage tolerance. At low temperatures, due to high surface energy, single element metallic nanowires such as Ag usually transform into a more preferred phase via nucleation and propagation of partial dislocation through the nanowire, enabling superplasticity. In high entropy alloy (HEA) CoNiCrFeMn nanowires, the motion of the partial dislocation is hindered by the friction due to difference in the lattice parameter of the constituent atoms which is responsible for the hardening and lowering the ductility. In this study, we have examined the temperature-dependent superplasticity of single component Ag and multicomponent CoNiCrFeMn HEA nanowires using molecular dynamics simulations. The results demonstrate that Ag nanowires exhibit apparent temperature-dependent superplasticity at cryogenic temperature due to (110) to (100) cross-section reorientation behavior. Interestingly, HEA nanowires can perform exceptional strength-ductility trade-offs at cryogenic temperatures. Even at high temperatures, HEA nanowires can still maintain good flow stress and ductility prior to failure. Mechanical properties of HEA nanowires are better than Ag nanowires due to synergistic interactions of deformation twinning, FCC-HCP phase transformation, and the special reorientation of the cross-section. Further examination reveals that simultaneous activation of twining induced plasticity and transformation induced plasticity are responsible for the plasticity at different stages and temperatures. These findings could be very useful for designing nanowires at different temperatures with high stability and superior mechanical properties in the semiconductor industry.
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