480 research outputs found
The mechanism of heat transfer in nucleate pool boiling
A criterion is developed for bubble initiation from a gas filled cavity on a surface in contact with a superheated layer of liquid. It is found that the temperature of bubble initiation on a given surface is a function of the temperature conditions in the liquid surrounding the cavity as well as the surface properties themselves. It is also found that the delay time between bubbles is a function of the bulk liquid temperature and the wall superheat and is not constant for a given surface. By consideration of the transient conduction into a layer of liquid on the surface, a thermal layer thickness is obtained. With this thickness and a critical wall superheat relation for the cavity, a bubble growth rate is obtained. Bubble departure is considered and it is found that the Jakob and Fritz relation works as long as the true (non-equilibrium) bubble contact angle is used. The effect on the departure size of the virtual mass in the surrounding liquid is found to be negligible at one gravity. That is, at one gravity the primary effect of bubble growth velocity on bubble departure size is found to be due to contact angle changes. The initiation, growth and departure criterions are each experimentally, individually, checked. They are then used to compute the heat transfer near the knee of the boiling curve using only an experimental determination of the number of bubbles as a function of wall superheat and other known quantities. Finally the q vs. Tw - Tsat relation is computed and measured and compared. The comparison is satisfactory.Office of Naval Research DS
Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductants
Protein folding homeostasis in the endoplasmic reticulum (ER) requires efficient protein thiol oxidation, but also relies on a parallel reductive process to edit disulfides during the maturation or degradation of secreted proteins. To critically examine the widely held assumption that reduced ER glutathione fuels disulfide reduction, we expressed a modified form of a cytosolic glutathione-degrading enzyme, ChaC1, in the ER lumen. ChaC1(CtoS) purged the ER of glutathione eliciting the expected kinetic defect in oxidation of an ER-localized glutathione-coupled Grx1-roGFP2 optical probe, but had no effect on the disulfide editing-dependent maturation of the LDL receptor or the reduction-dependent degradation of misfolded alpha-1 antitrypsin. Furthermore, glutathione depletion had no measurable effect on induction of the unfolded protein response (UPR); a sensitive measure of ER protein folding homeostasis. These findings challenge the importance of reduced ER glutathione and suggest the existence of alternative electron donor(s) that maintain the reductive capacity of the ER.Wellcome Trust [084812/Z/08/Z, 100140]; European Commission EU [277713]; Fundacao para a Ciencia e a Tecnologia [PTDC/QUI-BIQ/119677/2010]info:eu-repo/semantics/publishedVersio
Luminescent iridium complexes for detection of molybdate
Reactions of [Ir(CÅøN)2Cl]2 [HCÅøN = 2-(3-R-phenyl)pyridine, 2-(3-R-phenylpyrazole) R = H, Me] with Me2-phencat give luminescent complexes [Ir(CÅøN)2(Me2-phencat)][PF6] (Me2-2a, b, c)[PF6]. Deprotection of the methoxy groups with BBr3 is problematic as simultaneous bromination of the cyclometallated phenyl groups occurs. However, deprotection of Me2-phencat with BBr3 followed by complexation with [Ir(CÅøN)2Cl]2 gives luminescent complexes [Ir(CÅøN)2(H2-phencat)][PF6] (H2-3a, c)[PF6], which are luminescent sensors for molybdate
Visualizing RAD51-mediated joint molecules: implications for recombination mechanism and the effect of sequence heterology
The defining event in homologous recombination is the exchange of base-paired partners between a single-stranded (ss) DNA and a homologous duplex driven by recombinase proteins, such as human RAD51. To understand the mechanism of this essential genome maintenance event, we analyzed the structure of RAD51āDNA complexes representing strand exchange intermediates at nanometer resolution by scanning force microscopy. Joint molecules were formed between substrates with a defined ssDNA segment and homologous region on a double-stranded (ds) partner. We discovered and quantified several notable architectural features of RAD51 joint molecules. Each end of the RAD51-bound joints had a distinct structure. Using linear substrates, a 10-nt region of mispaired bases blocked extension of joint molecules in all examples observed, whereas 4ānt of heterology only partially blocked joint molecule extension. Joint molecules, including 10ānt of heterology, had paired DNA on either side of the heterologous substitution, indicating that pairing could initiate from the free 3ā²end of ssDNA or from a region adjacent to the ssāds junction. RAD51 filaments covering joint ssādsDNA regions were more stable to disassembly than filaments covering dsDNA. We discuss how distinct structural features of RAD51-bound DNA joints can play important roles as recognition sites for proteins that facilitate and control strand exchange
MAS NMR investigation of molecular order in an ionic liquid crystal
The structure and molecular order in the thermotropic ionic liquid crystal (ILC), [choline][geranate(H)octanoate], an analogue of Choline And GEranate (CAGE), which has potential for use as a broad-spectrum antimicrobial and transdermal and oral delivery agent, were investigated by magic-angle spinning (MAS) nuclear magnetic resonance (NMR), polarizing optical microscopy, small-angle X-ray scattering (SAXS), and mass spectrometry. Mass spectrometry and the 1H NMR chemical shift reveal that CAGE-oct is a dynamic system, with metathesis (the exchange of interacting ions) and hydrogen exchange occurring between hydrogen-bonded/ionic complexes such as [(choline)(geranate)(H)(octanoate)], [(choline)(octanoate)2(H)], and [(choline)(geranate)2(H)]. These clusters, which are shown by mass spectrometry to be significantly more stable than expected for typical electrostatic ion clusters, involve hydrogen bonding between the carboxylic acid, carboxylate, and hydroxyl groups, with rapid hydrogen bond breaking and re-formation observed to average the 1H chemical shifts. The formation of a partial bilayer liquid crystal (LC) phase was identified by SAXS and polarizing optical microscopy at temperatures below ā¼293 K. The occurrence of this transition close to room temperature could be utilized as a potential temperature-induced āswitchā of the anisotropic properties for particular applications. The presence of an isotropic component of approximately 23% was observed to coexist with the LC phase, as detected by polarizing optical microscopy and quantified by both 1Hā13C dipolar-chemical shift correlation (DIPSHIFT) and 1H double-quantum (DQ) MAS NMR experiments. At temperatures above the LC-to-isotropic transition, intermediate-range order (clustering of polar and nonpolar domains), a feature of many ILs, persists. Site-specific order parameters for the LC phase of CAGE-oct were obtained from the MAS NMR measurement of the partially averaged 13Cā1H dipolar couplings (DCH) by cross-polarization (CP) build-up curves and DIPSHIFT experiments, and 1Hā1H dipolar couplings (DHH) by double-quantum (DQ) build-up curves. The corresponding order parameters, SCH and SHH, are in the range 0ā0.2 and are lower compared to those for smectic (i.e., layered) phases of conventional nonionic liquid crystals, resembling those of lamellar phases formed by lyotropic surfactantāsolvent systems
Gene expression meta-analysis of Parkinsonās disease and its relationship with Alzheimerās disease
Abstract Parkinsonās disease (PD) and Alzheimerās disease (AD) are the most common neurodegenerative diseases and have been suggested to share common pathological and physiological links. Understanding the cross-talk between them could reveal potentials for the development of new strategies for early diagnosis and therapeutic intervention thus improving the quality of life of those affected. Here we have conducted a novel meta-analysis to identify differentially expressed genes (DEGs) in PD microarray datasets comprising 69 PD and 57 control brain samples which is the biggest cohort for such studies to date. Using identified DEGs, we performed pathway, upstream and protein-protein interaction analysis. We identified 1046 DEGs, of which a majority (739/1046) were downregulated in PD. YWHAZ and other genes coding 14ā3-3 proteins are identified as important DEGs in signaling pathways and in protein-protein interaction networks (PPIN). Perturbed pathways also include mitochondrial dysfunction and oxidative stress. There was a significant overlap in DEGs between PD and AD, and over 99% of these were differentially expressed in the same up or down direction across the diseases. REST was identified as an upstream regulator in both diseases. Our study demonstrates that PD and AD share significant common DEGs and pathways, and identifies novel genes, pathways and upstream regulators which may be important targets for therapy in both diseases
Rapid regulation of telomere length is mediated by poly(ADP-ribose) polymerase-1
Shelterin/telosome is a multi-protein complex at mammalian telomeres, anchored to the double-stranded region by the telomeric-repeat binding factors-1 and -2. In vitro modification of these proteins by poly(ADP-ribosyl)ation through poly(ADP-ribose) polymerases-5 (tankyrases) and -1/-2, respectively, impairs binding. Thereafter, at least telomeric-repeat binding factor-1 is degraded by the proteasome. We show that pharmacological inhibition of poly(ADP-ribose) polymerase activity in cells from two different species leads to rapid decrease in median telomere length and stabilization at a lower setting. Specific knockdown of poly(ADP-ribose) polymerase-1 by RNA interference had the same effect. The length of the single-stranded telomeric overhang as well as telomerase activity were not affected. Release of inhibition led to a fast re-gain in telomere length to control levels in cells expressing active telomerase. We conclude that poly(ADP-ribose) polymerase-1 activity and probably its interplay with telomeric-repeat binding factor-2 is an important determinant in telomere regulation. Our findings reinforce the link between poly(ADP-ribosyl)ation and aging/longevity and also impact on the use of poly(ADP-ribose) polymerase inhibitors in tumor therapy
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