Modulation of the slow/common gating of CLC channels by intracellular cadmium.

Members of the CLC family of Cl(-) channels and transporters are homodimeric integral membrane proteins. Two gating mechanisms control the opening and closing of Cl(-) channels in this family: fast gating, which regulates opening and closing of the individual pores in each subunit, and slow (or common) gating, which simultaneously controls gating of both subunits. Here, we found that intracellularly applied Cd(2+) reduces the current of CLC-0 because of its inhibition on the slow gating. We identified CLC-0 residues C229 and H231, located at the intracellular end of the transmembrane domain near the dimer interface, as the Cd(2+)-coordinating residues. The inhibition of the current of CLC-0 by Cd(2+) was greatly enhanced by mutation of I225W and V490W at the dimer interface. Biochemical experiments revealed that formation of a disulfide bond within this Cd(2+)-binding site is also affected by mutation of I225W and V490W, indicating that these two mutations alter the structure of the Cd(2+)-binding site. Kinetic studies showed that Cd(2+) inhibition appears to be state dependent, suggesting that structural rearrangements may occur in the CLC dimer interface during Cd(2+) modulation. Mutations of I290 and I556 of CLC-1, which correspond to I225 and V490 of CLC-0, respectively, have been shown previously to cause malfunction of CLC-1 Cl(-) channel by altering the common gating. Our experimental results suggest that mutations of the corresponding residues in CLC-0 change the subunit interaction and alter the slow gating of CLC-0. The effect of these mutations on modulations of slow gating of CLC channels by intracellular Cd(2+) likely depends on their alteration of subunit interactions

THERMAL BEHAVIOR IN A MULTI-LAYER METAL THIN-FILM WITH INTERFACIAL CONTACT CONDUCTANCE BY THE PARABOLIC TWO-STEP MODEL

ABSTRACT The effect of the contact conductance on micro-scale heat transport in a multi-layered metal thin-film subjected to ultrafast pulse laser heating is investigated in this study. The interfacial contact conductance existing at the interface is included in the analysis. The parabolic two-step (PTS) model is applied to predict the electron and lattice temperature. The equations can be solved by Laplace transform and the Riemann-sum approximation. The results show that the contact conductance plays an important role in the micro-scale heat transport processes for a multi-layered metal thin-film. Hence, in order to eliminate the thermal failure of the surface layer, the thermal resistance existing at the interface should be as small as possible. INTRODUCTION Ultra-fast pulse-laser heating on the metals are widely used in thermal processing of materials, for instance, laser micromachining, laser patterning, laser processing of diamond films from carbon ion implanted copper substrates, and laser surface hardening Since high-power laser can lead to thermal damage at the front surface of a single layer film, multi-layered metal films are widely used to avoid the problem of thermal damage [3] The effects of interfacial contact conductance and the thickness of the surface layer on the temperature solution of the material are also investigated. Lee and Tsai [10] also examined the parabolic two-step model t

Human parvovirus B19 nonstructural protein NS1 enhanced the expression of cleavage of 70 kDa U1-snRNP autoantigen

<p>Abstract</p> <p>Background</p> <p>Human parvovirus B19 (B19) is known to induce apoptosis that has been associated with a variety of autoimmune disorders. Although we have previously reported that B19 non-structural protein (NS1) induces mitochondrial-dependent apoptosis in COS-7 cells, the precise mechanism of B19-NS1 in developing autoimmunity is still obscure.</p> <p>Methods</p> <p>To further examine the effect of B19-NS1 in presence of autoantigens, COS-7 cells were transfected with pEGFP, pEGFP-B19-NS1 and pEGFP-NS1K334E, a mutant form of B19-NS1, and detected the expressions of autoantigens by various autoantibodies against Sm, U1 small nuclear ribonucleoprotein (U1-snRNP), SSA/Ro, SSB/La, Scl-70, Jo-1, Ku, and centromere protein (CENP) A/B by using Immunoblotting.</p> <p>Results</p> <p>Significantly increased apoptosis was detected in COS-7 cells transfected with pEGFP-B19-NS1 compared to those transfected with pEGFP. Meanwhile, the apoptotic 70 kDa U1-snRNP protein in COS-7 cells transfected with pEGFP-B19-NS1 is cleaved by caspase-3 and converted into a specific 40 kDa product, which were recognized by anti-U1-snRNP autoantibody. In contrast, significantly decreased apoptosis and cleaved 40 kDa product were observed in COS-7 cells transfected with pEGFP-NS1K334E compared to those transfected with pEGFP-B19-NS1.</p> <p>Conclusions</p> <p>These findings suggested crucial association of B19-NS1 in development of autoimmunity by inducing apoptosis and specific cleavage of 70 kDa U1-snRNP.</p

Surveillance cultures of samples obtained from biopsy channels and automated endoscope reprocessors after high-level disinfection of gastrointestinal endoscopes

BACKGROUND: The instrument channels of gastrointestinal (GI) endoscopes may be heavily contaminated with bacteria even after high-level disinfection (HLD). The British Society of Gastroenterology guidelines emphasize the benefits of manually brushing endoscope channels and using automated endoscope reprocessors (AERs) for disinfecting endoscopes. In this study, we aimed to assess the effectiveness of decontamination using reprocessors after HLD by comparing the cultured samples obtained from biopsy channels (BCs) of GI endoscopes and the internal surfaces of AERs. METHODS: We conducted a 5-year prospective study. Every month random consecutive sampling was carried out after a complete reprocessing cycle; 420 rinse and swabs samples were collected from BCs and internal surface of AERs, respectively. Of the 420 rinse samples collected from the BC of the GI endoscopes, 300 were obtained from the BCs of gastroscopes and 120 from BCs of colonoscopes. Samples were collected by flushing the BCs with sterile distilled water, and swabbing the residual water from the AERs after reprocessing. These samples were cultured to detect the presence of aerobic and anaerobic bacteria and mycobacteria. RESULTS: The number of culture-positive samples obtained from BCs (13.6%, 57/420) was significantly higher than that obtained from AERs (1.7%, 7/420). In addition, the number of culture-positive samples obtained from the BCs of gastroscopes (10.7%, 32/300) and colonoscopes (20.8%, 25/120) were significantly higher than that obtained from AER reprocess to gastroscopes (2.0%, 6/300) and AER reprocess to colonoscopes (0.8%, 1/120). CONCLUSIONS: Culturing rinse samples obtained from BCs provides a better indication of the effectiveness of the decontamination of GI endoscopes after HLD than culturing the swab samples obtained from the inner surfaces of AERs as the swab samples only indicate whether the AERs are free from microbial contamination or not

Influence of electrode thermal conductivity on resistive switching behavior during reset process

Resistive random access memory (RRAM) is the most promising candidate for non-volatile memory (NVM) due to its extremely low operation voltage, extremely fast write/erase speed, and excellent scaling capability. However, an obstacle hindering mass production of RRAM is the non-uniform physical mechanism in its resistance switching process. This study examines the influence of different electrode thermal conductivity on switching behavior during the reset process. Electrical analysis methods and an analysis of current conduction mechanism indicate that better thermal conductivity in the electrode will require larger input power in order to induce more active oxygen ions to take part in the reset process. More active oxygen ions cause a more complete reaction during the reset process, and cause the effective switching gap (dsw) to become thicker. The effect of the electrode thermal conductivity and input power are explained by our model and clarified by electrical analysis methods. Please click Additional Files below to see the full abstract

A case of acute appendicitis with Vibrio fluvialis peritonitis

AbstractHuman infections caused by Vibrio fluvialis are rarely reported. The most common clinical presentation of V. fluvialis infection is acute gastroenteritis with diarrhea. Reported extra-intestinal infections caused by V. fluvialis have included bacteremia, hemorrhagic cellulitis and cerebritis. Peritonitis is an uncommon clinical presentation of Vibrio infections, and most cases have occurred in patients receiving peritoneal dialysis or those with liver cirrhosis. Herein, we report the first case of acute appendicitis with V. fluvialis peritonitis

Cytotoxic Effect of Recombinant Mycobacterium tuberculosis CFP-10/ESAT-6 Protein on the Crucial Pathways of WI-38 Cells

To unravel the cytotoxic effect of the recombinant CFP-10/ESAT-6 protein (rCFES) on WI-38 cells, an integrative analysis approach, combining time-course microarray data and annotated pathway databases, was proposed with the emphasis on identifying the potentially crucial pathways. The potentially crucial pathways were selected based on a composite criterion characterizing the average significance and topological properties of important genes. The analysis results suggested that the regulatory effect of rCFES was at least involved in cell proliferation, cell motility, cell survival, and metabolisms of WI-38 cells. The survivability of WI-38 cells, in particular, was significantly decreased to 62% with 12.5 μM rCFES. Furthermore, the focal adhesion pathway was identified as the potentially most-crucial pathway and 58 of 65 important genes in this pathway were downregulated by rCFES treatment. Using qRT-PCR, we have confirmed the changes in the expression levels of LAMA4, PIK3R3, BIRC3, and NFKBIA, suggesting that these proteins may play an essential role in the cytotoxic process in the rCFES-treated WI-38 cells