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Xiaoyu Deng, Jiali Zhang, Jiazi Su, Hao Liu, Yanlong Cong, Lei Zhang, Kemeng Zhang, Ning Shi, Rongguang Lu, Xijun Yan
The aim of this study was to establish a multiplex PCR (mPCR) method that can simultaneously detect canine parvovirus (CPV-2), canine coronavirus (CCoV) and canine adenovirus (CAV), thereby eliminating the need to detect these pathogens individually. Based on conserved regions in the genomes of these three viruses, the VP2 gene of CPV-2, the endoribonuclease nsp15 gene of CCoV, and the 52K gene of CAV were selected for primer design. The specificity of the mPCR results showed no amplification of canine distemper virus (CDV), canine parainfluenza virus (CPIV), or pseudorabies virus (PRV), indicating that the method had good specificity...
April 19, 2018: Archives of Virology
Yaxiong Yang, Nan Liu, Yuanyuan He, Yuxia Liu, Lin Ge, Linzhi Zou, Sen Song, Wei Xiong, Xiaodong Liu
GCaMP, one popular type of genetically-encoded Ca2+ indicator, has been associated with various side-effects. Here we unveil the intrinsic problem prevailing over different versions and applications, showing that GCaMP containing CaM (calmodulin) interferes with both gating and signaling of L-type calcium channels (CaV 1). GCaMP acts as an impaired apoCaM and Ca2+ /CaM, both critical to CaV 1, which disrupts Ca2+ dynamics and gene expression. We then design and implement GCaMP-X, by incorporating an extra apoCaM-binding motif, effectively protecting CaV 1-dependent excitation-transcription coupling from perturbations...
April 17, 2018: Nature Communications
Zhenyu Hu, Guang Li, Jiong-Wei Wang, Suet Yen Chong, Dejie Yu, Xiaoyuan Wang, Jia Lin Soon, Mui Cheng Liang, Yuk Peng Wong, Na Huang, Henry M Colecraft, Ping Liao, Tuck Wah Soong
Background -L-type CaV 1.2 channels play crucial roles in regulation of blood pressure. Galectin-1 (Gal-1), has been reported to bind to the I-II loop of CaV 1.2 channels to reduce their current density. However, the mechanistic understanding for the down-regulation of CaV 1.2 channels by Gal-1, and whether Gal-1 plays a direct role in blood pressure regulation remain unclear. Methods - In vitro experiments involving co-IP, western blot, patch-clamp recordings, immunohistochemistry and pressure myography were used to evaluate the molecular mechanisms by which Gal-1 down-regulates CaV 1...
April 12, 2018: Circulation
Jihee Sohn, Hang Lin, Madalyn Rose Fritch, Rocky S Tuan
BACKGROUND: Adult mesenchymal stem cells (MSCs) are an important resource for tissue growth, repair, and regeneration. To utilize MSCs more effectively, a clear understanding of how they react to environmental cues is essential. Currently, relatively little is known about how the composition of the plasma membranes affects stem cell phenotype and properties. The presence of lipid molecules, including cholesterol in particular, in the plasma membrane plays a crucial role in regulating a variety of physiological processes in cells...
April 3, 2018: Stem Cell Research & Therapy
Wen Zeng, Jiansong Tang, Haicheng Li, Haixia Xu, Hongyun Lu, Hangya Peng, Chuwen Lin, Rili Gao, Shuo Lin, Keyi Lin, Kunying Liu, Yan Jiang, Jianping Weng, Longyi Zeng
Lipotoxicity leads to insulin secretion deficiency, which is among the important causes for the onset of type 2 diabetes mellitus. Thus, the restoration of β-cell mass and preservation of its endocrine function are long-sought goals in diabetes research. Previous studies have suggested that the membrane protein caveolin-1 (Cav-1) is implicated in β-cell apoptosis and insulin secretion, however, the underlying mechanisms still remains unclear. Our objective is to explore whether Cav-1 depletion protects pancreatic β cells from lipotoxicity and what are the underlying mechanisms...
March 26, 2018: Cellular Signalling
Han-Sen Chen, Xi Chen, Wen-Ting Li, Jian-Gang Shen
Reactive nitrogen species (RNS) play important roles in mediating cerebral ischemia-reperfusion injury. RNS activate multiple signaling pathways and participate in different cellular events in cerebral ischemia-reperfusion injury. Recent studies have indicated that caveolin-1 and matrix metalloproteinase (MMP) are important signaling molecules in the pathological process of ischemic brain injury. During cerebral ischemia-reperfusion, the production of nitric oxide (NO) and peroxynitrite (ONOO- ), two representative RNS, down-regulates the expression of caveolin-1 (Cav-1) and, in turn, further activates nitric oxide synthase (NOS) to promote RNS generation...
March 29, 2018: Acta Pharmacologica Sinica
Srinivasan Sundararaj, Dmitry Shishmarev, Yiechang Lin, Shouvik Aditya, Marco G Casarotto
Ahnak is a ~ 700 kDa polypeptide that was originally identified as a tumour-related nuclear phosphoprotein, but later recognized to play a variety of diverse physiological roles related to cell architecture and migration. A critical function of Ahnak is modulation of Ca2+ signaling in cardiomyocytes by interacting with the β subunit of the L-type Ca2+ channel (CaV 1.2). Previous studies have identified the C-terminal region of Ahnak, designated as P3 and P4 domains, as a key mediator of its functional activity...
March 28, 2018: Biomolecular NMR Assignments
Julie Briot, Olivier Mailhot, Benoîte Bourdin, Marie-Philippe Tétreault, Rafael Najmanovich, Lucie Parent
L-type CaV 1.2 channels are essential for the excitation-contraction coupling in cardiomyocytes and are hetero-oligomers of a pore-forming CaV α1C assembled with CaV β and CaV α2δ1 subunits. A direct interaction between CaV α2δ1 and Asp-181 in the first extracellular loop of CaV α1 reproduces the native properties of the channel. A 3-D model of the von Willebrand factor type A (VWA) domain of CaV α2δ1 complexed with the voltage sensor domain of CaV α1C suggests that Ser-261 and Ser-263 residues in the metal ion-dependent adhesion site (MIDAS) motif are determinant in this interaction, but this hypothesis is untested...
March 27, 2018: Journal of Biological Chemistry
Zhonglin Zhu, Zhilong Yu, Jianfeng Wang, Lisheng Zhou, Jing Zhang, Bin Yao, Jin Dou, Zhengjun Qiu, Chen Huang
BACKGROUND/AIMS: Krüppel-like factor 4 (KLF4), a member of the KLF family of zinc finger transcription factors, has been identified as a tumor suppressor gene in a variety of tumors. However, the molecular mechanisms by which KLF4 inhibits epithelial-to-mesenchymal transition (EMT) and metastasis in pancreatic cancer remain unclear. METHODS: KLF4 expression in pancreatic cancer was analyzed using public datasets (Oncomine and The Cancer Genome Atlas). The expression of KLF4, caveolin-1 (Cav-1), E-cadherin, and vimentin, and their correlations with clinicopathological characteristics were evaluated by immunohistochemistry in pancreatic cancer tissues...
March 21, 2018: Cellular Physiology and Biochemistry
Ke Wang, Xue Zhu, Yu Chen, Yongxiang Yin, Tao Ma
Metastatic triple-negative breast cancer (TNBC) has poor outcome with conventional chemotherapy regimens due to its aggressive behavior. The acquisition of anoikis resistance, a programmed cell death process triggered by substratum detachment, is an important mechanism in TNBC metastasis. Therefore, agents that can restore the sensitivity of cancer cells to anoikis may be helpful for the treatment of metastatic TNBC. In this study, we investigated the inhibitory effect of Tubeimosides V (TBMS-V), a cyclic bisdesmoside isolated from the ethanol extracts of tubers of B...
March 23, 2018: Archives of Biochemistry and Biophysics
Sascha Ra Alles, Esperanza Garcia, Sridhar Balasubramanyan, Karen Jones, John R Tyson, Twinkle Joy, Terrance P Snutch, Peter A Smith
Background Following peripheral nerve chronic constriction injury, the accumulation of the α2δ-1 auxiliary subunit of voltage-gated Ca2+ channels in primary afferent terminals contributes to the onset of neuropathic pain. Overexpression of α2δ-1 in Xenopus oocytes increases the opening properties of Cav 1.2 L-type channels and allows Ca2+ influx at physiological membrane potentials. We therefore posited that L-type channels play a role in neurotransmitter release in the superficial dorsal horn in the chronic constriction injury model of neuropathic pain...
January 2018: Molecular Pain
Johannes Goekler, Andreas Zuckermann, Alexandra Kaider, Philipp Angleitner, Emilio Osorio-Jaramillo, Roxana Moayedifar, Keziban Uyanik-Uenal, Frieda-Marie Kainz, Marco Masetti, Guenther Laufer, Arezu Z Aliabadi-Zuckermann
BACKGROUND: Evidence concerning an association between cytomegalovirus (CMV) infection and accelerated cardiac allograft vasculopathy (CAV) is inconclusive. METHODS: Data were analyzed retrospectively from 297 consecutive heart transplants between 1.1.2002 and 31.12.2012. Patients ≤ 18 years of age, survival and follow-up ≤ 1-year post-transplant and patients with early CAV were excluded. CMV-infection was diagnosed and monitored closely in the first year. CAV was diagnosed by coronary angiography via left heart catheterization and results were categorized according to the International Society of Heart and Lung Transplantation (ISHLT) scoring system...
March 26, 2018: Transplant International: Official Journal of the European Society for Organ Transplantation
Ana Sánchez, Cristina Contreras, Belén Climent, Alejandro Gutiérrez, Mercedes Muñoz, Albino García-Sacristán, Miguel López, Luis Rivera, Dolores Prieto
The impact of obesity on vascular smooth muscle (VSM) Ca2+ handling and vasoconstriction, and its regulation by the phosphatidylinositol 3-kinase (PI3K), mitogen activated protein kinase (MAPK) and protein kinase C (PKC) were assessed in mesenteric arteries (MA) from obese Zucker rats (OZR). Simultaneous measurements of intracellular Ca2+ ([Ca2+ ]i ) and tension were performed in MA from OZR and compared to lean Zucker rats (LZR), and the effects of selective inhibitors of PI3K, ERK-MAPK kinase and PKC were assessed on the functional responses of VSM voltage-dependent L-type Ca2+ channels (CaV 1...
March 21, 2018: Biochemical Pharmacology
Noémie Scheidel, Julie Kennedy, Oliver E Blacque
Primary cilium structure and function relies on control of ciliary membrane homeostasis, regulated by membrane trafficking processes that deliver and retrieve ciliary components at the periciliary membrane. However, the molecular mechanisms controlling ciliary membrane establishment and maintenance, especially in relation to endocytosis, remain poorly understood. Here, using Caenorhabditis elegans , we describe closely linked functions for early endosome (EE) maturation factors RABS-5 (Rabenosyn-5) and VPS-45 (VPS45) in regulating cilium length and morphology, ciliary and periciliary membrane volume, and ciliary signalling-related sensory behaviour...
March 23, 2018: EMBO Journal
Alessandra Vallone, Sarah D'Alessandro, Simone Brogi, Margherita Brindisi, Giulia Chemi, Gloria Alfano, Stefania Lamponi, Soon Goo Lee, Joseph M Jez, Karin J M Koolen, Koen J Dechering, Simona Saponara, Fabio Fusi, Beatrice Gorelli, Donatella Taramelli, Silvia Parapini, Reto Caldelari, Giuseppe Campiani, Sandra Gemma, Stefania Butini
Therapies addressing multiple stages of Plasmodium falciparum life cycle are highly desirable for implementing malaria elimination strategies. MMV019918 (1, 1-[5-(4-bromo-2-chlorophenyl)furan-2-yl]-N-[(piperidin-4-yl)methyl]methanamine) was selected from the MMV Malaria Box for its dual activity against both asexual stages and gametocytes. In-depth structure-activity relationship studies and cytotoxicity evaluation led to the selection of 25 for further biological investigation. The potential transmission blocking activity of 25 versus P...
March 10, 2018: European Journal of Medicinal Chemistry
Fang Yang, Guoxun Chen, Meihu Ma, Ning Qiu, Lingjiao Zhu, Jing Li
Phospholipids have been shown to modulate intestinal cholesterol absorption in cells and animals, a process that is regulated by several transporter proteins. Of these proteins, Niemann-Pick C1-Like 1 (NPC1L1) is a major contributor to this process. The mechanism by which phospholipids modulate cholesterol absorption remains unknown. Here, we evaluate the effects of egg-yolk phospholipids on cholesterol absorption and transport in human colon carcinoma cell line (Caco-2 cells) and on the expression of NPC1L1 and others proteins associated with cholesterol absorption (ABCG5, ABCG8, ABCA1, ACAT2, MTP, CAV-1, ANX-2)...
March 22, 2018: Lipids
Parmanand Malvi, Balkrishna Chaube, Shivendra Vikram Singh, Naoshad Mohammad, Maleppillil Vavachan Vijayakumar, Snahlata Singh, Surbhi Chouhan, Manoj Kumar Bhat
Background: Obesity is associated with increased risk, poor prognosis and outcome of therapy, in various cancers. Obesity-associated factors or adipokines, especially leptin and resistin, are purported to promote growth, survival, proliferation, and invasiveness of cancer cells. However, the mechanistic link between these adipokines and therapeutic response in malignancies is not clearly understood. Methods: ob/ob and db/db mouse models were used in this study to evaluate the role of leptin and resistin towards the outcome of dacarbazine (DTIC) therapy in melanoma...
2018: Cancer & Metabolism
Zhenlong Chen, Suellen D'Arc Oliveira, Adriana M Zimnicka, Ying Jiang, Tiffany Sharma, Stone Chen, Orly Lazarov, Marcelo G Bonini, Jacob M Haus, Richard D Minshall
We hypothesized that the maintenance of vascular homeostasis is critically dependent on the expression and reciprocal regulation of caveolin-1 (Cav-1) and endothelial nitric oxide synthase (eNOS) in endothelial cells (ECs). Skeletal muscle biopsies from subjects with type II diabetes showed 50% less Cav-1 and eNOS compared to lean healthy controls. The Cav-1:eNOS expression ratio was 200:1 in primary culture human ECs. Cav-1 siRNA reduced eNOS protein and gene expression in association with a 2-fold increase in eNOS phosphorylation and nitrate production per molecule of eNOS which was reversed in cells overexpressing Adv-Cav-1-GFP...
March 21, 2018: Molecular Biology of the Cell
Kirsten F Smit, Moritz Konkel, Raphaela Kerindongo, Maximilian A Landau, Coert J Zuurbier, Markus W Hollmann, Benedikt Preckel, Rienk Nieuwland, Martin Albrecht, Nina C Weber
Caveolins are involved in anaesthetic-induced cardioprotection. Actin filaments are located in close connection to Caveolins in the plasma membrane. We hypothesised that helium might affect the cytoskeleton and induce secretion of Caveolin. HCAEC, HUVEC and Cav-1 siRNA transfected HUVEC were exposed for 20 minutes to either helium (5% CO2 , 25% O2 , 70% He) or control gas (5% CO2 , 25% O2 , 70% N2 ). Cells and supernatants were collected for infrared Western blot analysis, immunofluorescence staining, nanoparticle tracking analysis and permeability measurements...
March 19, 2018: Scientific Reports
De-Gang Ji, Yan Zhang, Song-Mei Yao, Xu-Jie Zhai, Li-Rong Zhang, Yao-Zhong Zhang, Hui Li
Caveolin-1 (Cav-1), as a membrane protein involved in the formation of caveolae, binds steroid receptors and endothelial nitric oxide synthase, limiting its translocation and activation. In the present study, we investigated the role of Cav-1 in the progression of hepatic fibrosis induced by carbon tetrachloride (CCl4 ) in murine animals. Therefore, the wild type (WT) and Cav-1-knockout (Cav-1-/- ) mice were used in our study and subjected to CCl4 . The results indicated that CCl4 induced the decrease of Cav-1 expression in liver tissue samples...
March 14, 2018: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
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