Multiple roles of phosphoinositide-specific phospholipase C isozymes

Phosphoinositide-specific phospholipase C is an effector molecule in the signal transduction process. It generates two second messengers, inositol-1,4,5-trisphosphate and diacylglycerol from phosphatidylinositol 4,5-bisphosphate. Currently, thirteen mammal PLC isozymes have been identified, and they are divided into six groups: PLC-beta, -gamma, -delta, -epsilon, -zeta and -eta. Sequence analysis studies demonstrated that each isozyme has more than one alternative splicing variant. PLC isozymes contain the X and Y domains that are responsible for catalytic activity. Several other domains including the PH domain, the C2 domain and EF hand motifs are involved in various biological functions of PLC isozymes as signaling proteins. The distribution of PLC isozymes is tissue and organ specific. Recent studies on isolated cells and knockout mice depleted of PLC isozymes have revealed their distinct phenotypes. Given the specificity in distribution and cellular localization, it is clear that each PLC isozyme bears a unique function in the modulation of physiological responses. In this review, we discuss the structural organization, enzymatic properties and molecular diversity of PLC splicing variants and study functional and physiological roles of each isozyme.open19320

Interaction between integrin α5 and PDE4D regulates endothelial inflammatory signalling

Atherosclerosis is primarily a disease of lipid metabolism and inflammation; however, it is also closely associated with endothelial extracellular matrix (ECM) remodelling, with fibronectin accumulating in the laminin–collagen basement membrane. To investigate how fibronectin modulates inflammation in arteries, we replaced the cytoplasmic tail of the fibronectin receptor integrin α5 with that of the collagen/laminin receptor integrin α2. This chimaera suppressed inflammatory signalling in endothelial cells on fibronectin and in knock-in mice. Fibronectin promoted inflammation by suppressing anti-inflammatory cAMP. cAMP was activated through endothelial prostacyclin secretion; however, this was ECM-independent. Instead, cells on fibronectin suppressed cAMP via enhanced phosphodiesterase (PDE) activity, through direct binding of integrin α5 to phosphodiesterase-4D5 (PDE4D5), which induced PP2A-dependent dephosphorylation of PDE4D5 on the inhibitory site Ser651. In vivo knockdown of PDE4D5 inhibited inflammation at athero-prone sites. These data elucidate a molecular mechanism linking ECM remodelling and inflammation, thereby identifying a new class of therapeutic targets.United States. National Institutes of Health (5R01HL75092)National Science Foundation (U.S.). Materials Research Science and Engineering Centers (Program) (MR/J007412/1

Fatty acid compositions, free radical scavenging activities, and antioxidative enzyme activities of high-preference and low-preference beef cuts of Hanwoo () cows

Objective This study compared fatty acid compositions and antioxidant activities of high-preference cuts (loin, tenderloin, and rib) and low-preference cuts (brisket, topside, and shank) of Hanwoo (Bos taurus coreanae) cows to obtain potentially useful information for promoting the consumption of various low-preference cuts. Methods Individual 500 g samples of fresh beef were collected from each of the six cuts from 10 Hanwoo cows (quality grade 1) and immediately freeze-dried. The dried samples were evaluated for fatty acid composition, free radical scavenging activities (hydroxyl, alkyl, and 2, 2′-diphenyl-1-picrylhydrazyl [DPPH] radical), and antioxidative enzyme activities (glutathione peroxidase [GPx], glutathione-S-transferase [GST], and superoxide dismutase [SOD]). Results The percentages of total polyunsaturated fatty acids were significantly higher in low-preference cuts than in high-preference cuts (p<0.05). Hydroxyl, alkyl, and DPPH radical scavenging activities were significantly higher in low-preference cuts than in high-preference cuts (p<0.05). In addition, the activities of antioxidant enzymes, such as GPx, GST, and SOD, were significantly higher in low-preference cuts compared with high-preference cuts (p<0.05). Conclusion These results may influence consumers to include more low-preference cuts in their selections based on the nutritional facts, which could help to balance the beef market in South Korea

Role of Amphipathic Helix of a Herpesviral Protein in Membrane Deformation and T Cell Receptor Downregulation

Lipid rafts are membrane microdomains that function as platforms for signal transduction and membrane trafficking. Tyrosine kinase interacting protein (Tip) of T lymphotropic Herpesvirus saimiri (HVS) is targeted to lipid rafts in T cells and downregulates TCR and CD4 surface expression. Here, we report that the membrane-proximal amphipathic helix preceding Tip's transmembrane (TM) domain mediates lipid raft localization and membrane deformation. In turn, this motif directs Tip's lysosomal trafficking and selective TCR downregulation. The amphipathic helix binds to the negatively charged lipids and induces liposome tubulation, the TM domain mediates oligomerization, and cooperation of the membrane-proximal helix with the TM domain is sufficient for localization to lipid rafts and lysosomal compartments, especially the mutivesicular bodies. These findings suggest that the membrane-proximal amphipathic helix and TM domain provide HVS Tip with the unique ability to deform the cellular membranes in lipid rafts and to downregulate TCRs potentially through MVB formation

Functional characteristics of calcitonin gene-related peptide receptors in human Ewing's sarcoma WE-68 cells

AbstractCalcitonin gene-related peptide (CGRP) receptor activity was studied in WE-68 human Ewing's sarcoma cells. 125I-human CGRP bound in a time-dependent, reversible and saturable manner. Scatchard plots were compatible with the presence of a homogenous population of CGRP receptors with high affinity (Kd = 15 pM, and Bmax = 1.9 fmolmg protein). The potency order of unlabeled peptides, in the presence of radioligand, was: human CGRP-II > human CGRP = chick CGRP > rat CGRP = rat [Tyro]CGRP > human [Tyro] CGRP > > salmon calcitonin (CT) > rat [Tyro]CGRP-(28-37). Each peptide except CT and [Tyio]CGRP-(28-37) stimulated cyclic AMP generation in a concentration-dependent manner, and the relative potencies paralleled their relative ability in inhibiting 125I-human CGRP binding. We conclude that WE-68 Ewing's sarcoma cells express genuine CGRP receptors which upon activation lead to stimulation of cyclic AMP formation.Calcitonin gene-related peptide; Calcitonin; cyclic AMP; (Human; Ewing's sarcoma cell

Vascular Inflammatory Diseases and Endothelial Phenotypes

The physiological functions of endothelial cells control vascular tone, permeability, inflammation, and angiogenesis, which significantly help to maintain a healthy vascular system. Several cardiovascular diseases are characterized by endothelial cell activation or dysfunction triggered by external stimuli such as disturbed flow, hypoxia, growth factors, and cytokines in response to high levels of low-density lipoprotein and cholesterol, hypertension, diabetes, aging, drugs, and smoking. Increasing evidence suggests that uncontrolled proinflammatory signaling and further alteration in endothelial cell phenotypes such as barrier disruption, increased permeability, endothelial to mesenchymal transition (EndMT), and metabolic reprogramming further induce vascular diseases, and multiple studies are focusing on finding the pathways and mechanisms involved in it. This review highlights the main proinflammatory stimuli and their effects on endothelial cell function. In order to provide a rational direction for future research, we also compiled the most recent data regarding the impact of endothelial cell dysfunction on vascular diseases and potential targets that impede the pathogenic process

Integrins in mechanotransduction.

Forces acting on cells govern many important regulatory events during development, normal physiology, and disease processes. Integrin-mediated adhesions, which transmit forces between the extracellular matrix and the actin cytoskeleton, play a central role in transducing effects of forces to regulate cell functions. Recent work has led to major insights into the molecular mechanisms by which these adhesions respond to forces to control cellular signaling pathways. We briefly summarize effects of forces on organs, tissues, and cells; and then discuss recent advances toward understanding molecular mechanisms.info:eu-repo/semantics/publishe

Novel Modeling Approach to Analyze Threshold Voltage Variability in Short Gate-Length (15–22 nm) Nanowire FETs with Various Channel Diameters

In this study, threshold voltage (Vth) variability was investigated in silicon nanowire field-effect transistors (SNWFETs) with short gate-lengths of 15–22 nm and various channel diameters (DNW) of 7, 9, and 12 nm. Linear slope and nonzero y-intercept were observed in a Pelgrom plot of the standard deviation of Vth (σVth), which originated from random and process variations. Interestingly, the slope and y-intercept differed for each DNW, and σVth was the smallest at a median DNW of 9 nm. To analyze the observed DNW tendency of σVth, a novel modeling approach based on the error propagation law was proposed. The contribution of gate-metal work function, channel dopant concentration (Nch), and DNW variations (WFV, ∆Nch, and ∆DNW) to σVth were evaluated by directly fitting the developed model to measured σVth. As a result, WFV induced by metal gate granularity increased as channel area increases, and the slope of WFV in Pelgrom plot is similar to that of σVth. As DNW decreased, SNWFETs became robust to ∆Nch but vulnerable to ∆DNW. Consequently, the contribution of ∆DNW, WFV, and ∆Nch is dominant at DNW of 7 nm, 9 nm, and 12, respectively. The proposed model enables the quantifying of the contribution of various variation sources of Vth variation, and it is applicable to all SNWFETs with various LG and DNW

Inhibiting Integrin α5 Cytoplasmic Domain Signaling Reduces Atherosclerosis and Promotes Arteriogenesis

Background: Fibronectin in endothelial basement membranes promotes endothelial inflammatory activation and atherosclerosis but also promotes plaque stability and vascular remodeling. The fibronectin receptor α5 subunit is proinflammatory through binding to and activating phosphodiesterase 4D5, which inhibits anti‐inflammatory cyclic adenosine monophosphate and protein kinase A. Replacing the α5 cytoplasmic domain with that of α2 resulted in smaller atherosclerotic plaques. Here, we further assessed plaque phenotype and compensatory vascular remodeling in this model. Methods and Results: α5/2 mice in the hyperlipidemic apolipoprotein E null background had smaller plaques in the aortic root, with reduced endothelial NF‐κB activation and inflammatory gene expression, reduced leukocyte content, and much lower metalloproteinase expression. However, smooth muscle cell content, fibrous cap thickness, and fibrillar collagen were unchanged, indicating no shift toward vulnerability. In vivo knockdown of phosphodiesterase 4D5 also decreased endothelial inflammatory activation and atherosclerotic plaque size. α5/2 mice showed improved recovery from hindlimb ischemia after femoral artery ligation. Conclusions: Blocking the fibronectin‐Integrin α5 pathway reduces atherosclerotic plaque size, maintains plaque stability, and improves compensatory remodeling. This pathway is therefore a potential therapeutic target for treatment of atherosclerosis