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Lies Jordaens, Veerle Van Hoeck, Veronica Maillo, Alfonso Gutierrez-Adan, Waleed Fawzy A Marei, Bruno Vlaeminck, Sofie Thys, Roger G S Sturmey, Peter Bols, Jo Leroy
We hypothesized that elevated non-esterified fatty acids (NEFA) modify in vitro bovine oviduct epithelial cell (BOEC) metabolism and barrier function. Hereto, BOECs were studied in a polarized system with 24h-treatments at day 9: 1) CONTROL (0µM NEFA + 0%EtOH), 2) SOLVENT CONTROL (0µM NEFA + 0.45%EtOH), 3) BASAL NEFA (720µM NEFA + 0.45%EtOH in the basal compartment), 4) APICAL NEFA (720µM NEFA + 0.45%EtOH in the apical compartment). FITC-albumin was used for monolayer permeability assessment, and related to Transepithelial Electric Resistance (TER)...
March 3, 2017: Reproduction: the Official Journal of the Society for the Study of Fertility
Kristine E Olli, Kun Li, Deni S Galileo, Patricia A Martin-DeLeon
Reduced sperm motility (asthenospermia) and resulting infertility arise from deletion of the Plasma Membrane Ca(2+) -ATPase 4 (Pmca4) gene which encodes the highly conserved Ca(2+) efflux pump, PMCA4. This is the major Ca(2+) clearance protein in murine sperm. Since the mechanism underlying asthenospermia in PMCA4's absence or reduced activity is unknown, we investigated if sperm PMCA4 negatively regulates nitric oxide synthases (NOSs) and when absent NO, peroxynitrite, and oxidative stress levels are increased...
March 1, 2017: Journal of Cellular Physiology
M Kassan, A Vikram, Y R Kim, Q Li, A Kassan, H H Patel, S Kumar, M Gabani, J Liu, J S Jacobs, K Irani
Sirtuin1 (Sirt1) is a class III histone deacetylase that regulates a variety of physiological processes, including endothelial function. Caveolin1 (Cav1) is also an important determinant of endothelial function. We asked if Sirt1 governs endothelial Cav1 and endothelial function by regulating miR-204 expression and endoplasmic reticulum (ER) stress. Knockdown of Sirt1 in endothelial cells, and in vivo deletion of endothelial Sirt1, induced endothelial ER stress and miR-204 expression, reduced Cav1, and impaired endothelium-dependent vasorelaxation...
February 9, 2017: Scientific Reports
Soniya Charles, Vijay Raj, Jesu Arokiaraj, Kanchana Mala
Endothelial dysfunction (ED), an established response to cardiovascular risk factors, is characterized by increased levels of soluble molecules secreted by endothelial cells (EC). Evidence suggest that ED is an independent predictor of cardiac events and that it is associated with a deficiency in production or bioavailability of nitric oxide (NO) and/or an imbalance in the relative contribution of endothelium-derived relaxing and contracting factors. ED can be reversed by treating cardiovascular risk factors, hence, beyond ambiguity, ED contributes to initiation and progression of atherosclerotic disease...
May 2017: Pharmacological Research: the Official Journal of the Italian Pharmacological Society
James A Vassie, John M Whitelock, Megan S Lord
Cerium oxide nanoparticles (nanoceria) are widely reported to be cytocompatible and modulate intracellular reactive oxygen species (ROS) in a range of different cell types. In this study, nanoceria (d=7 and 94nm) synthesised by flame spray pyrolysis did not affect the proliferation of SKOV3 human ovarian and WiDr human colon cancer cell lines over a 72h treatment period. The cellular accumulation of nanoceria was uniform and increased up to 24h post-treatment before decreasing. The uptake of nanoceria in both cell lines was energy-dependent and was found to occur via non-specific pathways as well as clathrin-coated vesicles and caveolae...
December 8, 2016: Acta Biomaterialia
Raffaele Strippoli, Asier Echarri, Miguel Angel Del Pozo
Caveolin1, the main component of caveolae, plays a major role in regulating cell motility, gene expression, and cytoskeleton remodeling downstream of many membrane receptors. Here, we summarize different techniques set up to study changes in cell morphology and cell motility regulated by ERK/caveolin1 interactions during induction of epithelial mesenchymal transition (EMT) in mesothelial cells (MCs).
2017: Methods in Molecular Biology
Margaret E Tome, Joseph M Herndon, Charles P Schaefer, Leigh M Jacobs, Yifeng Zhang, Chelsea K Jarvis, Thomas P Davis
P-glycoprotein (PgP), a drug efflux pump in blood-brain barrier endothelial cells, is a major clinical obstacle for effective central nervous system drug delivery. Identifying PgP regulatory pathways that can be exploited clinically is critical for improving central nervous system drug delivery. We previously found that PgP activity increases in rat brain microvessels concomitant with decreased central nervous system drug delivery in response to acute peripheral inflammatory pain. In the current study, we tested the hypothesis that PgP traffics to the luminal plasma membrane of the microvessel endothelial cells from intracellular stores during peripheral inflammatory pain...
November 2016: Journal of Cerebral Blood Flow and Metabolism
Eliyahu Yosef, Adriana Katz, Yoav Peleg, Tevie Mehlman, Steven J D Karlish
Much evidence points to a role of Na,K-ATPase in ouabain-dependent signal transduction. Based on experiments with different cell lines and native tissue membranes, a current hypothesis postulates direct interactions between the Na,K-ATPase and Src kinase (non-receptor tyrosine kinase). Na,K-ATPase is proposed to bind Src kinase and inhibit its activity, whereas ouabain, the specific Na,K-ATPase inhibitor, binds and stabilizes the E2 conformation, thus exposing the Src kinase domain and its active site Tyr-418 for activation...
May 27, 2016: Journal of Biological Chemistry
Fang Xie, Shu-Hua Zhang, Jia Cheng, Hong-Wei Wang, Xiang Fei, Zi-Yu Jiao, Jie Tang, Yu-Kun Luo
We hypothesized that lack of the high-energy phosphates during liver storage may potentially cause persistent injury to the vascular endothelium. Biopsies were obtained from livers obtained from beating heart human donors, stored either in the standard storage solution, that is, University of Wisconsin solution (UWS) or Celsior, and examined for various markers related to progressive endothelial injury. The expression of P2Y1 receptor, the major signal transduction machinery for adenosine triphosphate/adenosine diphosphate, decreased in hepatic vascular endothelial cells over time...
January 2016: IUBMB Life
Isabelle Schrauwen, Szabolcs Szelinger, Ashley L Siniard, Ahmet Kurdoglu, Jason J Corneveaux, Ivana Malenica, Ryan Richholt, Guy Van Camp, Matt De Both, Shanker Swaminathan, Mari Turk, Keri Ramsey, David W Craig, Vinodh Narayanan, Matthew J Huentelman
A 3-year-old female patient presenting with an unknown syndrome of a neonatal progeroid appearance, lipodystrophy, pulmonary hypertension, cutis marmorata, feeding disorder and failure to thrive was investigated by whole-genome sequencing. This revealed a de novo, heterozygous, frame-shift mutation in the Caveolin1 gene (CAV1) (p.Phe160X). Mutations in CAV1, encoding the main component of the caveolae in plasma membranes, cause Berardinelli-Seip congenital lipodystrophy type 3 (BSCL). Although BSCL is recessive, heterozygous carriers either show a reduced phenotype of partial lipodystrophy, pulmonary hypertension, or no phenotype...
2015: PloS One
Shuohan Guo, Xiaohan Zhang, Mei Zheng, Xiaowei Zhang, Chengchun Min, Zengtao Wang, Seung Hoon Cheon, Min-Ho Oak, Seung-Yeol Nah, Kyeong-Man Kim
Among the multiple G protein-coupled receptor (GPCR) endocytic pathways, clathrin-mediated endocytosis (CME) and caveolar endocytosis are more extensively characterized than other endocytic pathways. A number of endocytic inhibitors have been used to block CME; however, systemic studies to determine the selectivity of these inhibitors are needed. Clathrin heavy chain or caveolin1-knockdown cells have been employed to determine the specificity of various chemical and molecular biological tools for CME and caveolar endocytosis...
October 2015: Biochimica et Biophysica Acta
Inbal Hazan-Halevy, Daniel Rosenblum, Shiri Weinstein, Osnat Bairey, Pia Raanani, Dan Peer
Mantle cell lymphoma (MCL) is an aggressive and incurable mature B cell neoplasm. The current treatments are based on chemotherapeutics and new class of drugs (e.g. Ibrutinib(®)), which in most cases ends with tumor resistance and relapse. Therefore, further development of novel therapeutic modalities is needed. Exosomes are natural extracellular vesicles, which play an important role in intercellular communication. The specificity of exosome uptake by different target cells remains unknown. In this study, we observed that MCL exosomes are taken up rapidly and preferentially by MCL cells...
August 1, 2015: Cancer Letters
Su Kil Jang, Jung Min Yu, Seung Tae Kim, Gwang Hoon Kim, Da Woon Park, Do Ik Lee, Seong Soo Joo
We demonstrated previously that ginsenoside Rg3 enhances the expression of macrophage scavenger receptor class A (SRA) and amyloid β peptide 1-42 (Aβ42) uptake in BV2 cells. In this study, we investigated the biochemical and mechanistic roles of Rg3 in human microglia and animal models to identify the determinants that participate in restoring memory and learning in brains disrupted by the Aβ42 peptide. SRA was expressed highly in Rg3-treated rats, and learning and memory functions were maintained at a normal level after the infusion of Aβ42...
July 5, 2015: European Journal of Pharmacology
Margaret E Tome, Charles P Schaefer, Leigh M Jacobs, Yifeng Zhang, Joseph M Herndon, Fabian O Matty, Thomas P Davis
Drug delivery to the brain for the treatment of pathologies with a CNS component is a significant clinical challenge. P-glycoprotein (PgP), a drug efflux pump in the endothelial cell membrane, is a major factor in preventing therapeutics from crossing the blood-brain barrier (BBB). Identifying PgP regulatory mechanisms is key to developing agents to modulate PgP activity. Previously, we found that PgP trafficking was altered concomitant with increased PgP activity and disassembly of high molecular weight PgP-containing complexes during acute peripheral inflammatory pain...
July 2015: Journal of Neurochemistry
Jagan Mohan, Björn Morén, Elin Larsson, Mikkel R Holst, Richard Lundmark
Caveolae are invaginations of the cell surface thought to regulate membrane tension, signalling, adhesion and lipid homeostasis owing to their dynamic behaviour ranging from stable surface association to dynamic rounds of fission and fusion with the plasma membrane. The caveolae coat is generated by oligomerisation of the membrane protein caveolin and the family of cavin proteins. Here, we show that cavin3 (also known as PRKCDBP) is targeted to caveolae by cavin1 (also known as PTRF) where it interacts with the scaffolding domain of caveolin1 and promote caveolae dynamics...
March 1, 2015: Journal of Cell Science
Masashi Isshiki
Endothelial cells regulates vascular tonus and exerts anti-atherosclerotic effects in response to external stimulation via Ca2+-dependent production of vasoactive substances such as NO. Agonists such as ATP hydrolyze membrane PIP2 and release soluble IP3 into the cell. IP3 then binds to the IP3 receptor and mobilizes Ca2+ from endoplasmic reticulum Ca2+ stores, followed by Ca2+ influxes from the extracellular space. Such store-operated Ca2+ entry comprises organization of a Ca2+ signal complex at the local subplasmalemmal domain involving TRPC4, caveolin1, and STIM1, a Ca2+ sensor protein for intracellular Ca2+ stores...
January 2015: Clinical Calcium
Shikha Kumari, Ashutosh Kumar, Puspendu Sardar, Manoj Yadav, Rakesh Kumar Majhi, Abhishek Kumar, Chandan Goswami
TRPV4 is involved in several physiological and sensory functions as well as with several diseases and genetic disorders, though the molecular mechanisms for these are unclear. In this work we have analyzed molecular evolution and structure-function relationship of TRPV4 using sequences from different species. TRPV4 has evolved during early vertebrate origin (450million years). Synteny analysis confirms that TRPV4 has coevolved with two enzymes involved in sterol biosynthesis, namely MVK and GLTP. Cholesterol-recognizing motifs are present within highly conserved TM4-Loop4-TM5 region of TRPV4...
January 2, 2015: Biochemical and Biophysical Research Communications
Bo Li, Zhiguo Zhang, Huizhi Zhang, Kai Quan, Yan Lu, Dongsheng Cai, Guang Ning
The prevalence of non-alcoholic fatty liver disease (NAFLD), a condition characterized by an excessive accumulation of triglycerides (TGs) in hepatocytes, has dramatically increased globally during recent decades. MicroRNAs (miRs) have been suggested to play crucial roles in many complex diseases and lipid metabolism. Our results indicated that miR199a-5p was remarkably upregulated in free fatty acid (FA)-treated hepatocytes. To investigate the role of miR199a-5p in the pathogenesis of fatty liver and the potential mechanism by which miR199a-5p regulates NAFLD, we first transfected two hepatocyte cell lines, HepG2 and AML12 cells, with agomiR199a-5p or antagomiR199a-5p...
December 2014: Journal of Molecular Endocrinology
Yeon Jeong Kim, Gi-Bang Koo, June-Young Lee, Hui-Sung Moon, Dong-Gun Kim, Da-Gyum Lee, Ju-Yeon Lee, Jin Ho Oh, Jong-Myeon Park, Minseok S Kim, Hyun Goo Woo, Seung-Il Kim, Pilsung Kang, Wonshik Choi, Tae Seok Sim, Woong-Yang Park, Jeong-Gun Lee, You-Sun Kim
Circulating tumor cells (CTCs) are rare cells and the presence of these cells may indicate a poor prognosis and a high potential for metastasis. Despite highly promising clinical applications, CTCs have not been investigated thoroughly, due to many technical limitations faced in their isolation and identification. Current CTC detection techniques mostly take the epithelial marker epithelial cell adhesion molecule (EpCAM), however, accumulating evidence suggests that CTCs show heterogeneous EpCAM expression due to the epithelial-to-mesenchymal transition (EMT)...
August 2014: Biomaterials
Sayaka Nagasawa, Kohei Ogura, Hiroyasu Tsutsuki, Hisako Saitoh, Joel Moss, Hirotaro Iwase, Masatoshi Noda, Kinnosuke Yahiro
The novel cytotoxic factor subtilase cytotoxin (SubAB) is produced mainly by non-O157 Shiga-toxigenic Escherichia coli (STEC). SubAB cleaves the molecular chaperone BiP/GRP78 in the endoplasmic reticulum (ER), leading to activation of RNA-dependent protein kinase (PKR)-like ER kinase (PERK), followed by caspase-dependent cell death. However, the SubAB uptake mechanism in HeLa cells is unknown. In this study, a variety of inhibitors and siRNAs were employed to characterize the SubAB uptake process. SubAB-induced BiP cleavage was inhibited by high concentrations of Dynasore, and methyl-β-cyclodextrin (mβCD) and Filipin III, but not suppressed in clathrin-, dynamin I/II-, caveolin1- and caveolin2-knockdown cells...
October 2014: Cellular Microbiology
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