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https://www.readbyqxmd.com/read/28324485/maintenance-of-human-embryonic-stem-cells-by-sphingosine-1-phosphate-and-platelet-derived-growth-factor
#1
Raymond C B Wong, Martin F Pera, Alice Pébay
Human embryonic stem cells (hESCs) have historically been cultivated on feeder layers of primary mouse embryonic fibroblasts (MEF) in a medium supplemented with fetal calf serum (FCS). However, serum contains a wide variety of biologically active compounds that might adversely affect hESC growth and differentiation. Thus, cultivation of stem cells in FCS complicates experimental approaches to define the intracellular mechanisms required for hESC maintenance. This chapter describes the serum-free maintenance of hESCs in culture by addition of sphingosine-1-phosphate (S1P) and platelet-derived growth factor (PDGF)...
March 22, 2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28324027/follistatin-targets-distinct-pathways-to-promote-brown-adipocyte-characteristics-in-brown-and-white-adipose-tissues
#2
Rajan Singh, Melissa Braga, Srinivasa T Reddy, Se-Jin Lee, Meher Parveen, Victor Grijalva, Laurent Vergnes, Shehla Pervin
We previously demonstrated that Fst expression is highest in brown adipose tissue (BAT) and skeletal muscle, but is also present at substantial levels in epididymal (Epi) and subcutaneous (SC) white adipose tissues (WAT). Fst promotes mouse brown preadipocyte differentiation and promotes browning during differentiation of mouse embryonic fibroblasts (MEFs). Fst-transgenic (Fst-Tg) mouse show significant increase in circulating Fst levels and increased brown adipose mass. BAT of Fst-Tg mice had increased expression of brown adipose-associated markers including uncoupling protein 1 (UCP1), PRDM16, PGC-1α and Glut4...
January 16, 2017: Endocrinology
https://www.readbyqxmd.com/read/28314909/localization-of-dynamin-related-protein-1-and-its-potential-role-in-lamellipodia-formation
#3
Youhwa Jo, Hyo Min Cho, Woong Sun, Jae Ryun Ryu
Dynamin-related protein1 (Drp1) plays an essential role in mitochondrial fission: Cytosolic Drp1 is translocated to the mitochondria upon stimulus, and oligomerized Drp1 constricts mitochondria by aid of actin filaments. Drp1 completes the fission process with GTP hydrolysis by its own GTPase activity. The importance of actin filament and its interaction with Drp1 in the mitochondrial fission process have been demonstrated. In this study, we found that Drp1 is enriched in the actin-rich leading edge of lamellipodia of mouse embryonic fibroblasts (MEFs) wherein mitochondria or peroxisomes are absent...
March 17, 2017: Histochemistry and Cell Biology
https://www.readbyqxmd.com/read/28314612/a-simple-imagej-macro-tool-for-analyzing-mitochondrial-network-morphology-in-mammalian-cell-culture
#4
Andrew J Valente, Lucas A Maddalena, Ellen L Robb, Fereshteh Moradi, Jeffrey A Stuart
Mitochondria exist in a dynamic cycle of fusion and fission whose balance directly influences the morphology of the 'mitochondrial network', a term that encompasses the branched, reticular structure of fused mitochondria as well as the separate, punctate individual organelles within a eukaryotic cell. Over the past decade, the significance of the mitochondrial network has been increasingly appreciated, motivating the development of various approaches to analyze it. Here, we describe the Mitochondrial Network Analysis (MiNA) toolset, a relatively simple pair of macros making use of existing ImageJ plug-ins, allowing for semi-automated analysis of mitochondrial networks in cultured mammalian cells...
March 14, 2017: Acta Histochemica
https://www.readbyqxmd.com/read/28306143/mutant-eif2b-leads-to-impaired-mitochondrial-oxidative-phosphorylation-in-vanishing-white-matter-disease
#5
Gali Raini, Reut Sharet, Melisa Herrero, Andrea Atzmon, Anjana Shenoy, Tamar Geiger, Orna Elroy-Stein
Eukaryotic translation initiation factor 2B (eIF2B) is a master regulator of protein synthesis under normal and stress conditions. Mutations in any of the five genes encoding its subunits lead to Vanishing White Matter (VWM) disease, a recessive genetic deadly illness caused by progressive loss of white matter in the brain. In the current study we used fibroblasts, which are not involved in the disease, to demonstrate the involvement of eIF2B in mitochondrial function and abundance. Mass-spectrometry of total proteome of mouse embryonic fibroblasts (MEFs) isolated from Eif2b5(R132H/R132H) mice revealed unbalanced stoichiometry of proteins involved in oxidative phosphorylation and of mitochondrial translation machinery components, among others...
March 17, 2017: Journal of Neurochemistry
https://www.readbyqxmd.com/read/28296014/geminin-is-an-indispensable-inhibitor-of-cdt1-in-mouse-embryonic-stem-cells
#6
Masaki Hosogane, Lena Bosu, Emiko Fukumoto, Hidetoshi Yamada, Soichiro Sato, Keiko Nakayama
Geminin is implicated in regulation of the cell cycle and differentiation. Although loss of Geminin triggers unscheduled DNA rereplication as a result of interruption of its interaction with Cdt1 in some somatic cancer cells, whether such cell cycle regulation also operates in embryonic stem cells (ESCs) has remained unclear. To characterize the Geminin-Cdt1 axis in ESCs and compare it with that in somatic cells, we established conditional knockout (KO) of Geminin in mouse ESCs and mouse embryonic fibroblasts (MEFs)...
March 14, 2017: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
https://www.readbyqxmd.com/read/28287329/atg4b-contains-a-c-terminal-lir-motif-important-for-binding-and-efficient-cleavage-of-mammalian-orthologs-of-yeast-atg8
#7
Mads Skytte Rasmussen, Stéphane Mouilleron, Birendra Kumar Shrestha, Martina Wirth, Rebecca Lee, Kenneth Bowitz Larsen, Yakubu Abudu Princely, Nicola O'Reilly, Eva Sjøttem, Sharon A Tooze, Trond Lamark, Terje Johansen
The cysteine protease ATG4B cleaves off one or more C-terminal residues of the inactive proform of proteins of the ortholog and paralog LC3 and GABARAP subfamilies of yeast Atg8 to expose a C-terminal glycine that is conjugated to phosphatidylethanolamine during autophagosome formation. We show that ATG4B contains a C-terminal LC3-interacting region (LIR) motif important for efficient binding to and cleavage of LC3 and GABARAP proteins. We solved the crystal structures of the GABARAPL1-ATG4B C-terminal LIR complex...
February 15, 2017: Autophagy
https://www.readbyqxmd.com/read/28287218/temperature-dependence-of-metal-enhanced-fluorescence-of-photosystem-i-from-thermosynechococcus-elongatus
#8
Imran Ashraf, Alexander Konrad, Heiko Lokstein, Sepideh Skandary, Michael Metzger, Joseph M Djouda, Thomas Maurer, Pierre M Adam, Alfred J Meixner, Marc Brecht
We report the temperature dependence of metal-enhanced fluorescence (MEF) of individual photosystem I (PSI) complexes from Thermosynechococcus elongatus (T. elongatus) coupled to gold nanoparticles (AuNPs). A strong temperature dependence of shape and intensity of the emission spectra is observed when PSI is coupled to AuNPs. For each temperature, the enhancement factor (EF) is calculated by comparing the intensity of individual AuNP-coupled PSI to the mean intensity of 'uncoupled' PSI. At cryogenic temperature (1...
March 13, 2017: Nanoscale
https://www.readbyqxmd.com/read/28286024/erk-induced-activation-of-tcf-family-of-srf-cofactors-initiates-a-chromatin-modification-cascade-associated-with-transcription
#9
Cyril Esnault, Francesco Gualdrini, Stuart Horswell, Gavin Kelly, Aengus Stewart, Phil East, Nik Matthews, Richard Treisman
We investigated the relationship among ERK signaling, histone modifications, and transcription factor activity, focusing on the ERK-regulated ternary complex factor family of SRF partner proteins. In MEFs, activation of ERK by TPA stimulation induced a common pattern of H3K9acS10ph, H4K16ac, H3K27ac, H3K9acK14ac, and H3K4me3 at hundreds of transcription start site (TSS) regions and remote regulatory sites. The magnitude of the increase in histone modification correlated well with changes in transcription. H3K9acS10ph preceded the other modifications...
March 16, 2017: Molecular Cell
https://www.readbyqxmd.com/read/28279564/pneumococcal-carriage-and-serotype-variation-before-and-after-introduction-of-pneumococcal-conjugate-vaccines-in-patients-with-acute-otitis-media-in-switzerland
#10
Aurélie Allemann, Pascal M Frey, Silvio D Brugger, Markus Hilty
BACKGROUND: Acute otitis media (AOM) is an important cause for antibiotic prescription within the paediatric population and Streptococcus pneumoniae is a major pathogen associated with AOM episodes. This study aimed at analysing the influence of the heptavalent and 13-valent pneumococcal conjugate vaccines (PCV7 and PCV13) on pneumococcal carriage and serotype distribution in AOM. METHODS: Nasopharyngeal swabs (NPS) and middle ear fluid (MEF) were collected within a Swiss surveillance study of outpatients from all ages with AOM between 2004 and 2015, covering three vaccination eras (pre-PCV7, PCV7 and PCV13)...
March 6, 2017: Vaccine
https://www.readbyqxmd.com/read/28270803/vibrio-cholerae-o1-with-reduced-susceptibility-to-ciprofloxacin-and-azithromycin-isolated-from-a-rural-coastal-area-of-bangladesh
#11
Shah M Rashed, Nur A Hasan, Munirul Alam, Abdus Sadique, Marzia Sultana, Md Mozammel Hoq, R Bradley Sack, Rita R Colwell, Anwar Huq
Cholera outbreaks occur each year in the remote coastal areas of Bangladesh and epidemiological surveillance and routine monitoring of cholera in these areas is challenging. In this study, a total of 97 Vibrio cholerae O1 isolates from Mathbaria, Bangladesh, collected during 2010 and 2014 were analyzed for phenotypic and genotypic traits, including antimicrobial susceptibility. Of the 97 isolates, 95 possessed CTX-phage mediated genes, ctxA, ace, and zot, and two lacked the cholera toxin gene, ctxA. Also both CTX(+) and CTX(-)V...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28270523/id1-promotes-obesity-by-suppressing-brown-adipose-thermogenesis-and-white-adipose-browning
#12
Mallikarjun Patil, Bal Krishan Sharma, Sawsan Elattar, Judith Chang, Shweta Kapil, Jinling Yuan, Ande Satyanarayana
Obesity results from increased energy intake or defects in energy expenditure. Brown adipose tissue (BAT) is specialized for energy expenditure, a process called adaptive thermogenesis. Peroxisome proliferator activated receptor γ coactivator 1α (PGC1α) controls BAT-mediated thermogenesis by regulating the expression of Ucp1 Inhibitor of differentiation 1 (Id1) is a helix-loop-helix transcription factor that plays important roles in cell proliferation and differentiation. Here, we demonstrate a novel function of Id1 in BAT thermogenesis and programming of beige adipocytes in white adipose tissue (WAT)...
March 7, 2017: Diabetes
https://www.readbyqxmd.com/read/28262793/characterization-of-cholesterol-homeostasis-in-sphingosine-1-phosphate-lyase-deficient-fibroblasts-reveals-a-niemann-pick-disease-type-c-like-phenotype-with-enhanced-lysosomal-ca-2-storage
#13
Hans Vienken, Nathalie Mabrouki, Katja Grabau, Ralf Frederik Claas, Agnes Rudowski, Nina Schömel, Josef Pfeilschifter, Dieter Lütjohann, Gerhild van Echten-Deckert, Dagmar Meyer Zu Heringdorf
Sphingosine-1-phosphate (S1P) lyase irreversibly cleaves S1P, thereby catalysing the ultimate step of sphingolipid degradation. We show here that embryonic fibroblasts from S1P lyase-deficient mice (Sgpl1(-/-)-MEFs), in which S1P and sphingosine accumulate, have features of Niemann-Pick disease type C (NPC) cells. In the presence of serum, overall cholesterol content was elevated in Sgpl1(-/-)-MEFs, due to upregulation of the LDL receptor and enhanced cholesterol uptake. Despite this, activation of sterol regulatory element-binding protein-2 was increased in Sgpl1(-/-)-MEFs, indicating a local lack of cholesterol at the ER...
March 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28259135/mir-155-is-positively-regulated-by-cbx7-in-mouse-embryonic-fibroblasts-and-colon-carcinomas-and-targets-the-kras-oncogene
#14
Floriana Forzati, Marco De Martino, Francesco Esposito, Romina Sepe, Simona Pellecchia, Umberto Malapelle, Gianluca Pellino, Claudio Arra, Alfredo Fusco
BACKGROUND: Loss of CBX7 expression has been described in several malignant neoplasias, including human colon and thyroid carcinomas proposing CBX7 as a tumor suppressor gene with a key role in cancer progression. This role is supported from the development of benign and malignant neoplasias in Cbx7 null mice. The aim of our work has been to investigate the mechanisms underlying the CBX7 oncosuppressor activity by analyzing the microRNAs (miRNAs) regulated by CBX7. METHODS: The miRNA expression profiles of the mouse embryonic fibroblasts (MEFs) null for Cbx7 and the wild-type counterpart were analyzed by the miRNACHIP microarray and then validated by qRT-PCR...
March 4, 2017: BMC Cancer
https://www.readbyqxmd.com/read/28257899/short-read-whole-genome-sequencing-for-determination-of-antimicrobial-resistance-mechanisms-and-capsular-serotypes-of-current-invasive-streptococcus-agalactiae-recovered-in-the-united-states
#15
Benjamin J Metcalf, Sopio Chochua, Robert E Gertz, Paulina A Hawkins, Jessica Ricaldi, Zhongya Li, Hollis Walker, Theresa Tran, Joy Rivers, Saundra Mathis, Delois Jackson, Anita Glennen, Ruth Lynfield, Lesley McGee, Bernard Beall
OBJECTIVES: Our objective was to evaluate and exploit a whole genome sequence (WGS) bioinformatics pipeline for predicting antimicrobial resistance and capsular serotypes from invasive group B streptococci (iGBS). METHODS: For 1975 iGBS recovered during 2015 from CDC's Active Bacterial Core surveillance, we compared pipeline predictions to broth dilution testing. Fifty-six isolates from earlier surveillance were included for testing β-lactams.. Conventional serotyping was compared to WGS-based assignments for 302 isolates...
February 28, 2017: Clinical Microbiology and Infection
https://www.readbyqxmd.com/read/28254433/loss-of-fkbp5-impedes-adipocyte-differentiation-under-both-normoxia-and-hypoxic-stress
#16
Lingling Zhang, Bin Qiu, Tingting Wang, Jun Wang, Ming Liu, Yuxue Xu, Chao Wang, Ran Deng, Kent Williams, Zhiwei Yang, Tiebing Liang, Weidong Yong
FK506-binding protein 51 (FKBP51) is one of the most important regulators in the GR-mediated stress response, and we previously demonstrated that loss of FKBP5 arrests adipogenesis and renders mice resistant to diet-induced obesity (DIO). However, the exact role of FKBP5 in the process of adipocyte differentiation under hypoxic conditions (the common microenvironment where adipocytes reside in obese individuals) is still unclear. Here, by isolating and culturing WT- and Fkbp5-knockout mouse embryonic fibroblasts (MEFs), and treat them at normal oxygen environment (21% O2, nomorxia) or low oxygen environment (5% O2, hypoxia)...
April 15, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28252045/latent-tgf-%C3%AE-binding-protein-2-and-4-have-essential-overlapping-functions-in-microfibril-development
#17
Yusuke Fujikawa, Hideyuki Yoshida, Tadashi Inoue, Tetsuya Ohbayashi, Kazuo Noda, Harald von Melchner, Toshiji Iwasaka, Ichiro Shiojima, Tomoya O Akama, Tomoyuki Nakamura
Microfibrils are exracellular matrix components necessary for elastic fiber assembly and for suspending lenses. We previously reported that latent TGF-β binding protein 2 (LTBP-2), a microfibril-associated protein, is required for forming stable microfibril bundles in ciliary zonules. However, it was not understood why Ltbp2 null mice only showed an eye-specific phenotype, whereas LTBP-2 is abundantly expressed in other tissues containing microfibrils in wild type mice. Here, we show that LTBP-4, another microfibril-associated protein, compensates for the loss of LTBP-2 in microfibril formation...
March 2, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28237928/robust-multi-exposure-image-fusion-a-structural-patch-decomposition-approach
#18
Kede Ma, Hui Li, Hongwei Yong, Zhou Wang, Deyu Meng, Lei Zhang
We propose a simple yet effective structural patch decomposition (SPD) approach for multi-exposure image fusion (MEF) that is robust to ghosting effect. We decompose an image patch into three conceptually independent components: signal strength, signal structure, and mean intensity. Upon fusing these three components separately, we reconstruct a desired patch and place it back into the fused image. This novel patch decomposition approach benefits MEF in many aspects. First, as opposed to most pixel-wise MEF methods, the proposed algorithm does not require post-processing steps to improve visual quality or to reduce spatial artifacts...
February 20, 2017: IEEE Transactions on Image Processing: a Publication of the IEEE Signal Processing Society
https://www.readbyqxmd.com/read/28231656/-acute-effect-of-ambient-air-pollution-on-small-airway-lung-functions-among-school-children-in-shanghai
#19
L J Zhang, C Y Guo, H H Xu, D Xu, X B Shen, X Y Du, M H Zhang, J G Tan, J H Zhang, C Y Dong, H L Qian, Y W Shi, M Z Pan, X D Zhou
Objective: To study the acute effects of compound ambient air pollution on small airway lung functions among school children in Shanghai. Method: A longitudinal survey on lung functions was conducted among 233 school-children from three schools (A, B and C, located in innerring, mid-ring and outer-ring areas). Lung function test was performed once a week for 3 times respectively, among children in school A and B in Dec. 2013 and in school C in Dec. 2014. The fourth lung function test was tested in Jun. 2014 and May 2015 in the respective schools...
February 10, 2017: Zhonghua Liu Xing Bing Xue za Zhi, Zhonghua Liuxingbingxue Zazhi
https://www.readbyqxmd.com/read/28217805/-induction-of-robust-senescence-associated-secretory-phenotype-in-mouse-nih-3t3-cells-by-mitomycin-c
#20
Wei-Xing Huang, Xiao-Xuan Guo, Zhong-Zhi Peng, Chun-Liang Weng, Chun-Yan Huang, Ben-Yan Shi, Jie Yang, Xiao-Xin Liao, Xiao-Yi Li, Hui-Ling Zheng, Xin-Guang Liu, Xue-Rong Sun
Senescence-associated secretory phenotype (SASP) is often a concomitant result of cell senescence, embodied by the enhanced function of secretion. The SASP factors secreted by senescent cells include cytokines, proteases and chemokines, etc, which can exert great influence on local as well as systemic environment and participate in the process of cell senescence, immunoregulation, angiogenesis, cell proliferation and tumor invasion, etc. Relative to the abundance of SASP models in human cells, the in vitro SASP model derived from mouse cells is scarce at present...
February 25, 2017: Sheng Li Xue Bao: [Acta Physiologica Sinica]
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