Chk1 is an evolutionarily conserved protein kinase that regulates cell cycle progression in response to checkpoint activation.In this study, we demonstrated agents block DNA replication or cause certain forms of damage induce the phosphorylation human Chk1.The phosphorylated form possessed higher intrinsic activity and eluted more quickly on gel filtration columns.Serines 317 345 were identified as sites vivo, ATR (the ATM-and Rad3-related kinase) both these vitro.Furthermore, serines vivo...

Checkpoint kinase (Chk)1 is an evolutionarily conserved protein that was first identified in fission yeast as essential component of the DNA damage checkpoint. In mice, Chk1 provides function absence environmentally imposed genotoxic stress. Here we show human cells lacking exhibit defects both ionizing radiation (IR)-induced S and G 2 checkpoints. addition, loss resulted accumulation a hypophosphorylated form Cdc25A phosphatase, Chk1-deficient failed to degrade after IR. The IR-induced...

Human placenta estradiol 17 beta-dehydrogenase (E2DH) cDNA clones were isolated from a lambda gt11 expression library by screening with 33 mer synthetic oligonucleotides derived the amino acid sequence of catalytic site E2DH and polyclonal antibodies raised against enzyme purified human placenta. Using 32P-labeled fragments coding 5'-untranslated regions, two mRNA species have been identified in poly(A)+ RNA placenta, major migrating at 1.3 kilobases (kb) while minor one is found 2.2 kb....

The mechanism underlying enhancer-blocking by insulators is unclear. We explored the activity of human beta-globin HS5, orthologue CTCF-dependent chicken HS4 insulator. An extra copy HS5 placed between locus control region (LCR) and downstream genes on a transgene fulfills classic predictions for an enhancer-blocker. Ectopic does not perturb LCR but blocks gene activation interfering with RNA pol II, activator coactivator recruitment, epigenetic modification at gene. Underlying these...

Chromatid catenation is actively monitored in human cells, with progression from G 2 to mitosis being inhibited when chromatids are insufficiently decatenated. Mitotic delay was quantified normal and checkpoint-deficient cells during treatment ICRF-193, a topoisomerase II catalytic inhibitor that prevents chromatid decatenation without producing topoisomerase-associated DNA strand breaks. Ataxia telangiectasia (A-T) defective damage checkpoints, showed mitotic treated ICRF-193. The response...

Regulation of the renal Na/H exchanger NHE-3 by protein kinase A (PKA) is a key intermediate step in hormonal regulation acid-base and salt balance. We studied role phosphorylation this process NHE-deficient AP-1 cells transfected with OKP expressing native NHE-3. dominant-negative PKA-regulatory subunit completely abolished effect cAMP on activity demonstrating PKA functional cAMP. isolated from cAMP-treated showed lower purified PKAin vitro suggesting that substrate vivo. Although changes...

Parathyroid hormone (PTH) is a potent inhibitor of mammalian renal proximal tubule Na(+) transport via its action on the apical membrane Na(+)/H(+) exchanger NHE3. In opossum kidney cell line, inhibition NHE3 activity was detected from 5 to 45 min after PTH addition. Increase in phosphorylation multiple serines evident PTH, but decrease surface antigen not detectable until 30 PTH. The due increased endocytosis. When endocytic trafficking arrested with dominant negative dynamin mutant (K44A),...

Purpose To determine whether HIV-1 produces microRNAs and elucidate these miRNAs can induce inflammatory response in macrophages (independent of the conventional miRNA function RNA interference) leading to chronic immune activation. Methods Using sensitive quantitative Real Time RT-PCR sequencing, we detected novel HIV-derived sera HIV+ persons, associated with exosomes. Release TNFα by challenged HIV was measured ELISA. Results infection primary alveolar produced elevated levels viral...

Abstract Background To investigate whether the administration of hydrogen/oxygen mixture was superior to oxygen in improving symptoms patients with acute exacerbation chronic obstructive pulmonary disease (AECOPD). Methods This prospective, randomized, double-blind, controlled clinical trial 10 centres enrolled patient AECOPD and a Breathlessness, Cough, Sputum Scale (BCSS) score at least 6 points. Eligible were randomly assigned (in 1:1 ratio) receive either or therapy. Primary endpoint...

ABSTRACT Type 1 fimbriae, expressed by most Escherichia coli strains, are thought to attach human uroepithelium as an initial step in the pathogenesis of urinary tract infections (UTI). Numerous reports using both vitro and murine models support this role for type fimbriae colonization. Unfortunately, only a limited number studies have directly examined expression vivo. To determine whether fimbrial genes transcribed during acute UTI, we employed modification established method. The...

ABSTRACT The virulence of a urease-negative mutant uropathogenic Proteus mirabilis and its wild-type parent strain was assessed by using CBA mouse model catheterized urinary tract infection. Overall, mice were significantly more susceptible than uncatheterized to infection P. . At high inoculum, the successfully colonized bladders but did not cause urolithiasis still severely attenuated in ability ascend kidneys. Using confocal laser scanning microscopy electron microscopy, we demonstrated...

The triglyceride–glucose (TyG) index, an emerging surrogate marker of insulin resistance, has been implicated in adverse cardiovascular outcomes. However, its prognostic value critically ill patients with atrial fibrillation (AF) remains unclear. This study aimed to investigate the association between TyG index and all-cause mortality this high-risk population. We identified AF from Medical Information Mart for Intensive Care IV (MIMIC-IV) database categorized them into tertiles based on...

Proteus mirabilis , associated with complicated urinary tract infection, expresses mannose‐resistant/Proteus‐like (MR/P) fimbriae. Expression of these surface structures, which mediate haemagglutination and have a demonstrated role in virulence, undergoes phase variation. By DNA sequence analysis, 252 bp invertible element was found the intergenic region between mrpI putative site‐specific recombinase gene, mrpA primary structural subunit gene. The segment is flanked by identical 21 inverted...

Endothelial cell (EC) apoptosis contributes to cigarette smoke (CS)-induced pulmonary emphysema. Metabolism of glucose, glutamine, and fatty acid is dysregulated in patients with chronic obstructive disease (COPD). Whether CS causes metabolic dysregulation ECs leading development COPD remains elusive. We hypothesized that alters metabolism, resulting lung ECs. To test this hypothesis, we treated primary mouse microvascular (PMVECs) extract (CSE) employed PMVECs from healthy subjects...

Summary: Proteus mirabilis, a motile Gram-negative bacterium, is principal cause of urinary tract infections in patients with functional or anatomical abnormalities the those catheters place. Thus far, virulence factors including urease, flagella, haemolysin, various fimbriae, IgA protease and deaminase have been characterized based on phenotypic traits conferred by these proteins. In this study, an attempt was made to identify new genes P. mirabilis that may not identifiable phenotypes...

Whereas the SLAMF-associated protein (SAP) is involved in differentiation of T follicular helper (Tfh) cells and antibody responses, precise requirements SLAMF receptors humoral immune responses are incompletely understood. By analyzing mice with targeted disruptions Slamf1, Slamf5, Slamf6 genes, we found that both T-dependent T-independent were twofold higher compared to those single knockout mice. These data suggest a suppressive synergy SLAMF1, SLAMF5, SLAMF6 immunity, which contrasts...