A1F-B, a novel CCAAT-binding transcription activator that interacts with the aldolase B promoter

AbstractWe describe here a 70 kDa transcription factor A1F-B, which preferentially binds to an element encompassing a CCAAT motif on the rat aldolase B promoter. Comparison of binding specificities, relative molecular masses, and subunit compositions with those of other known CCAAT-binding factors indicated that A1F-B is a novel member of CCAAT-binding factors

Selective labeling of a single organelle by using two-photon conversion of a photoconvertible fluorescent protein

SPIE BiOS, 2008, San Jose, California, United StatesWataru Watanabe, Tomoko Shimada, Sachihiro Matsunaga, Daisuke Kurihara, Shin-ichi Arimura, Nobuhiro Tsutsumi, Kiichi Fukui, Kazuyoshi Itoh, "Selective labeling of a single organelle by using two-photon conversion of a photoconvertible fluorescent protein," Proc. SPIE 6860, Multiphoton Microscopy in the Biomedical Sciences VIII, 68601B (15 February 2008); https://doi.org/10.1117/12.768745

Electrical muscle stimulation on upper and lower limb muscles in critically ill patients

Objectives: Electrical muscle stimulation (EMS) is widely used to enhance lower limb mobilization. Although upper limb muscle atrophy is common in critically ill patients, EMS application for the upper limbs has been rarely reported. The purpose of this study was to investigate whether EMS prevents upper and lower limb muscle atrophy and improves physical function. Design: Randomized controlled trial. Setting: Two-center, mixed medical/surgical intensive care unit (ICU). Patients: Adult patients who were expected to be mechanically ventilated for >48 h and stay in the ICU for >5 days. Interventions: Forty-two patients were randomly assigned to the EMS (n = 17) or control group (n = 19). Measurements and Main Results: Primary outcomes were change in muscle thickness and cross-sectional area of the biceps brachii and rectus femoris from day 1 to 5. Secondary outcomes included incidence of ICU-acquired weakness (ICU-AW), ICU mobility scale (IMS), length of hospitalization, and amino acid levels. The change in biceps brachii muscle thickness was −1.9% vs. −11.2% in the EMS and control (p = 0.007) groups, and the change in cross-sectional area was −2.7% vs. −10.0% (p = 0.03). The change in rectus femoris muscle thickness was −0.9% vs. −14.7% (p = 0.003) and cross-sectional area was −1.7% vs. −10.4% (p = 0.04). No significant difference was found in ICU-AW (13% vs. 40%; p = 0.20) and IMS (3 vs. 2; p = 0.42) between the groups. The length of hospitalization was shorter in the EMS group (23 [19–34] vs. 40 [26–64] days) (p = 0.04). On day 3, the change in the branched-chain amino acid level was lower in the EMS group (40.5% vs. 71.5%; p = 0.04). Conclusion: In critically ill patients, EMS prevented upper and lower limb muscle atrophy and attenuated proteolysis and decreased the length of hospitalization

Bioorganic synthesis of a recombinant HIV-1 fusion inhibitor, SC35EK, with an N-terminal pyroglutamate capping group.

The bioorganic synthesis of an end-capped anti-HIV peptide from a recombinant protein was investigated. Cyanogen bromide-mediated cleavage of two Met-Gln sites across the target anti-HIV sequence generated an HIV-1 fusion inhibitor (SC35EK) analog bearing an N-terminal pyroglutamate (pGlu) residue and a C-terminal homoserine lactone (Hsl) residue. The end-capped peptide, pGlu-SC35EK-Hsl, had similar bioactivity and biophysical properties to the parent peptide, and an improved resistance to peptidase-mediated degradation was observed compared with the non-end-capped peptide obtained using standard recombinant technology

Minimal important changes in standard deviation units are highly variable and no universally applicable value can be determined

[Objectives] This study aims to describe the distribution of anchor-based minimal important change (MIC) estimates in standard deviation (SD) units and examine if the robustness of such estimates depends on the specific SD used or on the methodological credibility of the anchor-based estimates. [Design and Setting] We included all anchor-based MIC estimates from studies published in MEDLINE and relevant literature databases upto October 2018. Each MIC was converted to SD units using baseline, endpoint, and change from baseline SDs. We performed a descriptive analysis of MICs in SD units and checked how the distribution would change if MICs with low methodological credibility were excluded from the analysis. [Results] We included 1, 009 MIC estimates from 182 studies. The medians and interquartile ranges of MICs in SD units were 0.43 (0.25 to 0.69), 0.42 (0.22 to 0.70), and 0.51 (0.28 to 0.78) for baseline, endpoint, and change SD units, respectively. Some MICs were extremely large or small. The distribution did not change significantly after excluding MICs estimated by less credible methods. [Conclusions] The size of the universally applicable MIC in SD units could not be determined. Anchor-based MICs in SD units were widely distributed, with more than half in the range of 0.2 to 0.8

Cryo-EM structures of human zinc transporter ZnT7 reveal the mechanism of Zn²⁺ uptake into the Golgi apparatus

クライオ電子顕微鏡により、ゴルジ体の亜鉛輸送体による亜鉛輸送機構の全容を解明 細胞の亜鉛恒常性維持機構の理解に大きな進展. 京都大学プレスリリース. 2023-08-29.Zinc ions (Zn²⁺) are vital to most cells, with the intracellular concentrations of Zn²⁺ being tightly regulated by multiple zinc transporters located at the plasma and organelle membranes. We herein present the 2.2-3.1 Å-resolution cryo-EM structures of a Golgi-localized human Zn²⁺/H+ antiporter ZnT7 (hZnT7) in Zn²⁺-bound and unbound forms. Cryo-EM analyses show that hZnT7 exists as a dimer via tight interactions in both the cytosolic and transmembrane (TM) domains of two protomers, each of which contains a single Zn²⁺-binding site in its TM domain. hZnT7 undergoes a TM-helix rearrangement to create a negatively charged cytosolic cavity for Zn²⁺ entry in the inward-facing conformation and widens the luminal cavity for Zn²⁺ release in the outward-facing conformation. An exceptionally long cytosolic histidine-rich loop characteristic of hZnT7 binds two Zn²⁺ ions, seemingly facilitating Zn²⁺ recruitment to the TM metal transport pathway. These structures permit mechanisms of hZnT7-mediated Zn²⁺ uptake into the Golgi to be proposed

Potential Neuroprotective Effects of an LSD1 Inhibitor in Retinal Ganglion Cells via p38 MAPK Activity

Citation: Tsutsumi T, Iwao K, Hayashi H, et al. Potential neuroprotective effects of an LSD1 inhibitor in retinal ganglion cells via p38 MAPK activity. Invest Ophthalmol Vis Sci. 2016;57:6461-6473. DOI:10.1167/ iovs.16-19494 PURPOSE. The epigenetic mechanisms associated with ocular neurodegenerative diseases remain unclear. The present study aimed to determine the role of lysine-specific demethylase 1 (LSD1), which represses transcription by removing the methyl group from methylated lysine 4 of histone H3, in retinal ganglion cell (RGC) survival, and to investigate the details of the neuroprotective mechanism of tranylcypromine, a major LSD1 inhibitor. METHODS. The authors evaluated whether tranylcypromine contributes to neuronal survival following stress-induced damage using primary cultured rat RGCs and in vivo N-methyl-Daspartate (NMDA)-induced excitotoxicity. Additionally, the molecules associated with tranylcypromine treatment were assessed by microarray and immunoblot analysis. RESULTS. Tranylcypromine significantly suppressed neuronal cell death following glutamate neurotoxicity and oxidative stress. Microarray and immunoblot analyses revealed that p38 mitogen-activated protein kinase (MAPK)c was a key molecule involved in the neuroprotective mechanisms induced by tranylcypromine because the significant suppression of p38 MAPKc by glutamate was reversed by tranylcypromine. Moreover, although pharmacologic inhibition of the phosphorylation of the total p38 MAPKs interfered with neuroprotective effects of tranylcypromine, the specific inhibition of p38 MAPKa and p38 MAPKb did not influence RGC survival. This suggests that the non-p38 MAPKa/b isoforms have important roles in neuronal survival by tranylcypromine. Additionally, the intravitreal administration of tranylcypromine significantly saved RGC numbers in an in vivo glaucoma model employing NMDA-induced excitotoxicity. CONCLUSIONS. These findings indicate that tranylcypromine-induced transcriptional and epigenetic regulation modulated RGC survival via the promotion of p38 MAPKc activity. Therefore, pharmacologic treatments that suppress LSD1 activity may be a novel therapeutic strategy that can be used to treat neurodegenerative diseases