Label-free detection of DNA methylation could be achieved using Raman spectroscopy with discriminant analysis.

Acidification is observed in stroke ranging between pH 6.1 and 6.5 in focal and global ischemia. This acidification induces deleterious effects including release of protease and calcium, activation of acid-sensing ion channels, mitochondrial destruction, and more. The pathological aspects of acidification have been well documented; nonetheless our understanding of how pH contributes to neuronal death in cerebral ischemia is still incomplete. Here, we examined the vulnerability of NBCn1 knockout mice to neuronal death induced by brain insults. Compared to wild-type littermates, NBCn1 knockout mice develop low plasma and CSF pH. To determined neuroprotective effect, mice were administered with NMDA (75 mg/kg body weight; intraperitoneal injection; 8 males for each group of knockouts and wild types). Knockout mice showed substantially reduced expression and activity of active caspase-3, and low TUNEL staining. Thus, knockout mice exhibit reduced neuronal death. In cultures of hippocampal neurons from postnat...

A cDNA encoding a double-stranded-RNA (dsRNA)-binding protein was isolated by screening a HeLa cell cDNA expression library for proteins that bind the HIV-1 Rev-responsive-element RNA. The cDNA encoded a protein that was identical to TRBP, the previously reported cellular protein that binds the transactivation response element (TAR) RNA of human immunodeficiency virus type 1. TRBP inhibited phosphorylation of the interferon-induced ribosome-associated protein kinase PKR and of the eukaryotic translation initiation factor eIF-2 alpha in a transient-expression system in which the translation of a reporter gene was inhibited by the localized activation of PKR. TRBP expression in HeLa cells complemented the growth and protein-synthesis defect of a vaccinia virus mutant lacking the expression of the dsRNA-binding protein E3L. These results implicate TRBP as a cellular regulatory protein that binds RNAs containing specific secondary structure(s) to mediate the inhibition of PKR activation and stimulate translation in a localized manner.

Supplementary Material 1.

Supplementary Material 1.

Abstract Calcium ions (Ca²⁺) are ubiquitous signaling molecules that play important roles as messengers controlling a variety of cellular functions in eukaryotes. Intracellular Ca²⁺ is tightly regulated by a complex interplay between channels, pumps, and exchangers. Aging disrupts Ca²⁺ homeostasis, contributing to age-related diseases, such as neurodegenerative disorders. Previous studies have highlighted the role of Ca²⁺ in regulating actin cytoskeletal proteins in human dermal fibroblasts (HDFs). In this study, we investigated the effect of age on Ca²⁺ influx and efflux in HDFs to test the hypothesis that age-related perturbations in Ca²⁺ homeostasis are correlated with increased F-actin content in aged HDFs. We observed that aging leads to quantitative and functional changes in Ca²⁺ channels, resulting in differences in Ca²⁺ dynamics. Older HDFs showed reduced Ca²⁺ influx and accelerated outflow compared to neonatal and young cells, potentially inhibiting gelsolin activity and increasing F-actin content. Although Ca²⁺ alone cannot fully explain the age-dependent F-actin changes, our findings suggest that it plays an important role in cellular senescence.

Abstract Calcium ions (Ca²⁺) are ubiquitous signaling molecules that play important roles as messengers controlling a variety of cellular functions in eukaryotes. Intracellular Ca²⁺ is tightly regulated by a complex interplay between channels, pumps, and exchangers. Aging disrupts Ca²⁺ homeostasis, contributing to age-related diseases, such as neurodegenerative disorders. Previous studies have highlighted the role of Ca²⁺ in regulating actin cytoskeletal proteins in human dermal fibroblasts (HDFs). In this study, we investigated the effect of age on Ca²⁺ influx and efflux in HDFs to test the hypothesis that age-related perturbations in Ca²⁺ homeostasis are correlated with increased F-actin content in aged HDFs. We observed that aging leads to quantitative and functional changes in Ca²⁺ channels, resulting in differences in Ca²⁺ dynamics. Older HDFs showed reduced Ca²⁺ influx and accelerated outflow compared to neonatal and young cells, potentially inhibiting gelsolin activity and increasing F-actin content. Although Ca²⁺ alone cannot fully explain the age-dependent F-actin changes, our findings suggest that it plays an important role in cellular senescence.

Label-free detection of DNA methylation could be achieved using Raman spectroscopy with discriminant analysis.

Acidification is observed in stroke ranging between pH 6.1 and 6.5 in focal and global ischemia. This acidification induces deleterious effects including release of protease and calcium, activation of acid-sensing ion channels, mitochondrial destruction, and more. The pathological aspects of acidification have been well documented; nonetheless our understanding of how pH contributes to neuronal death in cerebral ischemia is still incomplete. Here, we examined the vulnerability of NBCn1 knockout mice to neuronal death induced by brain insults. Compared to wild-type littermates, NBCn1 knockout mice develop low plasma and CSF pH. To determined neuroprotective effect, mice were administered with NMDA (75 mg/kg body weight; intraperitoneal injection; 8 males for each group of knockouts and wild types). Knockout mice showed substantially reduced expression and activity of active caspase-3, and low TUNEL staining. Thus, knockout mice exhibit reduced neuronal death. In cultures of hippocampal neurons from postnat...