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Antiretroviral therapy (ART) can lower a patient’s HIV plasma viral load to an undetectable level, but following cessation of ART viremia rapidly rebounds. It has been shown that ART does not eliminate latent viruses sequestered into anatomical and cellular reservoirs. Therefore, in patients that have ceased ART, the following rebound in HIV viremia is caused by the activation of latent HIV reservoirs. A major issue in HIV cure research is the quantification of these latent HIV reservoirs. Various reservoir measurement methods exist, but the gold standard technique remains the culture-based quantitative viral outgrowth assay (QVOA). Recently, a new PCR-based assay, named the tat/rev induced limiting dilution assay (TILDA) was described which measures the frequency of inducible latently infected CD4+ T cells that actively produce multiply-spliced RNA coding for the Tat/Rev proteins. The objective of this study was to further optimize the assay by examining the influence of varied factors, such as the amount of products transferred from the pre-amplification step to the PCR reaction, storage of pre-amplification products prior to PCR runs, and the number of cells used, on the assay’s sensitivity and reproducibility. We also investigated whether the assay could be used to quantify HIV reservoirs in monocytes/macrophages. [ABSTRACT FROM AUTHOR]

Mammary gland development during late pregnancy in sows is a major factor affecting the composition of colostrum and milk and the pre-weaning growth of piglets, while valine is essential for protein and nitrogen metabolism in mammary gland of sow. However, the effects of valine and its underlying mechanism on mammary gland development during late pregnancy are still unclear. Here, we hypothesized that dosage of dietary valine during late pregnancy will affect protein synthesis of colostrum in gilts. The results showed that supplementation of valine during late pregnancy significantly increased content of protein (P < 0.01), fat (P = 0.02) and solids-non-fat (P = 0.04) in colostrum. Our in vitro study also confirmed that valine supplementation increased protein synthesis and cell proliferation in porcine mammary epithelial cells (PMEC). Furthermore, these changes were associated with elevated phosphorylation levels of mammalian target of rapamycin (mTOR), and ribosomal protein S6 kinase (S6) and eukaryotic initiation factor 4E-binding protein-1 (4EBP1) in valine-supplemented cells, which could be effectively blocked by the antagonists of mTOR. These findings indicated that valine enhanced mammary gland development and protein synthesis in colostrum via the mTOR signaling pathway. These results, using an in vivo and in vitro model, helped to understand the beneficial effects of dietary valine supplementation on gilts. [ABSTRACT FROM AUTHOR]