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https://www.readbyqxmd.com/read/29791858/a-multi-layered-quantitative-in-vivo-expression-atlas-of-the-podocyte-unravels-kidney-disease-candidate-genes
#1
Markus M Rinschen, Markus Gödel, Florian Grahammer, Stefan Zschiedrich, Martin Helmstädter, Oliver Kretz, Mostafa Zarei, Daniela A Braun, Sebastian Dittrich, Caroline Pahmeyer, Patricia Schroder, Carolin Teetzen, HeonYung Gee, Ghaleb Daouk, Martin Pohl, Elisa Kuhn, Bernhard Schermer, Victoria Küttner, Melanie Boerries, Hauke Busch, Mario Schiffer, Carsten Bergmann, Marcus Krüger, Friedhelm Hildebrandt, Joern Dengjel, Thomas Benzing, Tobias B Huber
Damage to and loss of glomerular podocytes has been identified as the culprit lesion in progressive kidney diseases. Here, we combine mass spectrometry-based proteomics with mRNA sequencing, bioinformatics, and hypothesis-driven studies to provide a comprehensive and quantitative map of mammalian podocytes that identifies unanticipated signaling pathways. Comparison of the in vivo datasets with proteomics data from podocyte cell cultures showed a limited value of available cell culture models. Moreover, in vivo stable isotope labeling by amino acids uncovered surprisingly rapid synthesis of mitochondrial proteins under steady-state conditions that was perturbed under autophagy-deficient, disease-susceptible conditions...
May 22, 2018: Cell Reports
https://www.readbyqxmd.com/read/29784858/production-of-staphylococcal-complement-inhibitor-scin-and-other-immune-modulators-during-the-early-stages-of-staphylococcus-aureus-biofilm-formation-in-a-mammalian-cell-culture-medium
#2
Andi R Sultan, Jasper W Swierstra, Nicole A Lemmens-den Toom, Susan V Snijders, Silvie Hansenová Maňásková, Annelies Verbon, Willem J B van Wamel
Immune modulators are known to be produced by matured biofilms and during different stages of planktonic growth of Staphylococcus aureus. Little is known about immune modulator production during the early stages of biofilm formation, thus raising the question: how does S. aureus protect itself from the innate immune responses at these stages? Therefore, we determined the production of the following immune modulators: chemotaxis inhibitory protein of staphylococci (CHIPS), staphylococcal complement inhibitor (SCIN), formyl peptide receptor-like 1 inhibitor, gamma-hemolysin component B, leukocidin D, E, and S, staphylococcal superantigen-like protein 1, 3, 5 and 9, and staphylococcal enterotoxin A...
May 21, 2018: Infection and Immunity
https://www.readbyqxmd.com/read/29784826/deletion-of-lrrtm1-and-lrrtm2-in-adult-mice-impairs-basal-ampa-receptor-transmission-and-ltp-in-hippocampal-ca1-pyramidal-neurons
#3
Mehdi Bhouri, Wade Morishita, Paul Temkin, Debanjan Goswami, Hiroshi Kawabe, Nils Brose, Thomas C Südhof, Ann Marie Craig, Tabrez J Siddiqui, Robert Malenka
Leucine-rich repeat transmembrane (LRRTM) proteins are synaptic cell adhesion molecules that influence synapse formation and function. They are genetically associated with neuropsychiatric disorders, and via their synaptic actions likely regulate the establishment and function of neural circuits in the mammalian brain. Here, we take advantage of the generation of a LRRTM1 and LRRTM2 double conditional knockout mouse ( LRRTM1,2 cKO) to examine the role of LRRTM1,2 at mature excitatory synapses in hippocampal CA1 pyramidal neurons...
May 21, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29784672/genetic-screening-and-multipotency-in-rhesus-monkey-haploid-neural-progenitor-cells
#4
Haisong Wang, Wenhao Zhang, Jian Yu, Congyu Wu, Qian Gao, Xu Li, Yanni Li, Jinxin Zhang, Yaru Tian, Tao Tan, Weizhi Ji, Luyuan Li, Yang Yu, Ling Shuai
Haploid embryonic stem cells (haESCs) have been extensively applied in forward and reverse genetic screening. However, the mammalian haploid somatic cell line is difficult to achieve because of spontaneous diploidization in differentiation. As a non-human primate species, monkeys are widely used in basic and pre-clinical research in which haploid cells are restricted to ESCs. Here, we report that rhesus monkey haESCs in an optimized culture medium showed naïve-state pluripotency and stable haploidy. This model facilitated the derivation of haploid neural progenitor cells (haNPCs), which maintained haploidy and differentiation potential into neurons and glia for a long period in vitro High-throughput trapping mutations can be efficiently introduced into haNPCs via piggyBac transposons...
May 21, 2018: Development
https://www.readbyqxmd.com/read/29782020/whole-mount-immunofluorescence-and-follicle-quantification-of-cultured-mouse-ovaries
#5
Vera D Rinaldi, Jordana C Bloom, John C Schimenti
Research in the field of mammalian reproductive biology often involves evaluating the overall health of ovaries and testes. Specifically, in females, ovarian fitness is often assessed by visualizing and quantifying follicles and oocytes. Because the ovary is an opaque three-dimensional tissue, traditional approaches require laboriously slicing the tissue into numerous serial sections in order to visualize cells throughout the entire organ. Furthermore, because quantification by this method typically entails scoring only a subset of the sections separated by the approximate diameter of an oocyte, it is prone to inaccuracy...
May 2, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29781726/in-vitro-analysis-of-glucose-and-quercetin-effects-on-m-tor-and-nrf-2-expression-in-hepg2-cell-line-diabetes-and-cancer-connection
#6
Amir Yarahmadi, Fatemeh Khademi, Zohreh Mostafavi-Pour, Fatemeh Zal
Some types of cancers show a strong relationship with diabetes and play a central role in mortality in the patient population suffering from diabetes mellitus. In this study, HepG2 cells have been used to investigate the toxic effects of hyperglycemia and/or quercetin (Q) on mammalian target of rapamycin (m-TOR) and nuclear factor erythroid 2-related factor 2 (Nrf-2) expression as central molecules involved in cancer. HepG2 cells were cultured with different concentrations of glucose (5.5, 30, and 50 mM) and/or Q (25 µM) for 48 and 72 h...
May 21, 2018: Nutrition and Cancer
https://www.readbyqxmd.com/read/29781256/improved-performance-in-mammalian-cell-perfusion-cultures-by-growth-inhibition
#7
Moritz K F Wolf, Aurélie Closet, Monika Bzowska, Jean-Marc Bielser, Jonathan Souquet, Hervé Broly, Massimo Morbidelli
Mammalian cell perfusion cultures represent a promising alternative for the production of various biopharmaceuticals to the current fed-batch technology. Long-term operation at a fixed viable cell density (VCD) requires a viable culture and a constant removal of excessive cells. Product loss in the cell removing bleed stream deteriorates the process yield. In this study, we investigated the use of chemical and environmental growth inhibition on culture performance by either adding valeric acid (VA) to the production media or by reducing the culture temperature (33...
May 21, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29781033/microarray-analysis-of-circular-rna-expression-profiles-associated-with-gemcitabine-resistance-in-pancreatic-cancer-cells
#8
Chao Xu, Yue Yu, Fei Ding
Pancreatic cancer (PC) is one of the most malignant tumors of the digestive system due to its rapid progression, metastasis and resistance to chemotherapy. Gemcitabine (GEM) chemotherapy is the first‑choice treatment for advanced PC. However, the effect of GEM‑based chemotherapy on PC is limited due to the development of chemoresistance, and the molecular mechanisms underlying this resistance have yet to be investigated. Circular RNAs (circRNAs), which can function as microRNA sponges, have been found to be involved in the development of several types of cancer...
May 17, 2018: Oncology Reports
https://www.readbyqxmd.com/read/29776129/assay-based-on-electrical-impedance-spectroscopy-to-discriminate-between-normal-and-cancerous-mammalian-cells
#9
Fabián Eduardo Giana, Fabián José Bonetto, Mariela Inés Bellotti
In this work we present an assay to discriminate between normal and cancerous cells. The method is based on the measurement of electrical impedance spectra of in vitro cell cultures. We developed a protocol consisting on four consecutive measurement phases, each of them designed to obtain different information about the cell cultures. Through the analysis of the measured data, 26 characteristic features were obtained for both cell types. From the complete set of features, we selected the most relevant in terms of their discriminant capacity by means of conventional statistical tests...
March 2018: Physical Review. E
https://www.readbyqxmd.com/read/29772587/anti-inflammatory-effects-of-cardamonin-in-ovarian-cancer-cells-are-mediated-via-mtor-suppression
#10
Huajiao Chen, Daohua Shi, Peiguang Niu, Yanting Zhu, Jintuo Zhou
Cardamonin exhibits a variety of pharmacological activities including anti-inflammatory and antitumor, which are correlated with the inhibition of nuclear factor-kappaB and the mammalian target of rapamycin, respectively. However, whether the anti-inflammatory effects of cardamonin are mediated by the mammalian target of rapamycin remains unknown. In this study, ovarian cancer SKOV3 cells were cultured with lipopolysaccharide to induce inflammation, and the inhibitory effects and underlying molecular mechanisms of cardamonin were investigated using specific inhibitors of the mammalian target of rapamycin and the nuclear factor-kappaB pathway (rapamycin and pyrrolidine dithiocarbamate, respectively)...
May 17, 2018: Planta Medica
https://www.readbyqxmd.com/read/29772025/shape-shifting-trypanosomes-flagellar-shortening-followed-by-asymmetric-division-in-trypanosoma-congolense-from-the-tsetse-proventriculus
#11
Lori Peacock, Christopher Kay, Mick Bailey, Wendy Gibson
Trypanosomatids such as Leishmania and Trypanosoma are digenetic, single-celled, parasitic flagellates that undergo complex life cycles involving morphological and metabolic changes to fit them for survival in different environments within their mammalian and insect hosts. According to current consensus, asymmetric division enables trypanosomatids to achieve the major morphological rearrangements associated with transition between developmental stages. Contrary to this view, here we show that the African trypanosome Trypanosoma congolense, an important livestock pathogen, undergoes extensive cell remodelling, involving shortening of the cell body and flagellum, during its transition from free-swimming proventricular forms to attached epimastigotes in vitro...
May 2018: PLoS Pathogens
https://www.readbyqxmd.com/read/29771971/constitutive-metanephric-mesenchyme-specific-expression-of-interferon-gamma-causes-renal-dysplasia-by-regulating-sall1-expression
#12
Kangsun Yun, Arthur A Hurwitz, Alan O Perantoni
Transplacental viral and parasitic infections have been shown to initiate an innate response in the mammalian embryo by increasing the expression of pro-inflammatory cytokines such as interferon-gamma (Ifng). However, the developmental consequences of an activated innate immunity and, in particular, the effects of induction of Ifng expression independent of infection have been largely overlooked. Here, we demonstrate in vivo that the conditional overexpression of Ifng in metanephric mesenchymal (MM) progenitors results in renal agenesis or hypoplasia...
2018: PloS One
https://www.readbyqxmd.com/read/29770355/host-regulated-hepatitis-b-virus-capsid-assembly-in-a-mammalian-cell-free-system
#13
Kuancheng Liu, Jianming Hu
The hepatitis B virus (HBV) is an important global human pathogen and represents a major cause of hepatitis, liver cirrhosis and liver cancer. The HBV capsid is composed of multiple copies of a single viral protein, the capsid or core protein (HBc), plays multiple roles in the viral life cycle, and has emerged recently as a major target for developing antiviral therapies against HBV infection. Although several systems have been developed to study HBV capsid assembly, including heterologous overexpression systems like bacteria and insect cells, in vitro assembly using purified protein, and mammalian cell culture systems, the requirement for non-physiological concentrations of HBc and salts and the difficulty in manipulating host regulators of assembly presents major limitations for detailed studies on capsid assembly under physiologically relevant conditions...
April 20, 2018: Bio-protocol
https://www.readbyqxmd.com/read/29769334/reovirus-nonstructural-protein-%C3%AF-ns-acts-as-an-rna-stability-factor-promoting-viral-genome-replication
#14
Paula F Zamora, Liya Hu, Jonathan J Knowlton, Roni M Lahr, Rodolfo A Moreno, Andrea J Berman, B V Venkataram Prasad, Terence S Dermody
Viral nonstructural proteins, which are not packaged into virions, are essential for replication of most viruses. Reovirus, a nonenveloped, double-stranded RNA (dsRNA) virus, encodes three nonstructural proteins that are required for viral replication and dissemination in the host. Reovirus nonstructural protein σNS is a single-stranded RNA (ssRNA)-binding protein that must be expressed in infected cells for production of viral progeny. However, activities of σNS during individual steps of the reovirus replication cycle are poorly understood...
May 16, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29762713/snap23-kif5-complex-controls-mglu1-receptor-trafficking
#15
Fabrice Raynaud, Vincent Homburger, Martial Seveno, Oana Vigy, Enora Moutin, Laurent Fagni, Julie Perroy
Metabotropic glutamate receptors are expressed at excitatory synapses and control synaptic transmission in mammalian brain. These receptors are involved in numerous patho-physiological functions. However, little is known about the molecular determinants responsible for their intracellular transport and membrane targeting. Here we investigated the nature of the molecular motor and adaptor protein responsible for trafficking and membrane localization of the group I metabotropic glutamate mGlu1 post-synaptic receptor in cultured hippocampal neurons...
May 14, 2018: Journal of Molecular Cell Biology
https://www.readbyqxmd.com/read/29757989/synthesis-and-evaluation-of-antiplasmodial-activity-of-2-2-2-trifluoroethoxychalcones-and-2-fluoroethoxy-chalcones-against-plasmodium-falciparum-in-culture
#16
Kavita Devi, Vinoth Rajendran, T M Rangarajan, Rishi Pal Singh, Prahlad C Ghosh, Manjula Singh
A new class of compounds comprising two series of chalcones with 2,2,2-trifluoroethoxy group and 2-fluoroethoxy groups were synthesized and screened for in vitro antiplasmodial activity against Plasmodium falciparum (3D7) using the [³H] hypoxanthine incorporation inhibition assay. Chalcones with 2,2,2-trifluoroethoxy groups substituted on the p - and m -positions of the 1-phenyl ring showed weak antiplasmodial activity, while compounds substituted on the o -position of the 1-phenyl ring displayed enhanced antiplasmodial activity, thus indicating that 2,2,2-trifluoroethoxy groups on the 1-phenyl ring of chalcones show position-dependent antiplasmodial activity...
May 14, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/29757290/the-lambda-select-cii-mutation-detection-system
#17
Ahmad Besaratinia, Stella Tommasi
A number of transgenic animal models and mutation detection systems have been developed for mutagenicity testing of carcinogens in mammalian cells. Of these, transgenic mice and the Lambda (λ) Select cII Mutation Detection System have been employed for mutagenicity experiments by many research groups worldwide. Here, we describe a detailed protocol for the Lambda Select cII mutation assay, which can be applied to cultured cells of transgenic mice/rats or the corresponding animals treated with a chemical/physical agent of interest...
April 26, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29754876/bone-morphogenetic-protein-4-antagonizes-hair-cell-regeneration-in-the-avian-auditory-epithelium
#18
Rebecca M Lewis, Jesse J Keller, Liangcai Wan, Jennifer S Stone
Permanent hearing loss is often a result of damage to cochlear hair cells, which mammals are unable to regenerate. Non-mammalian vertebrates such as birds replace damaged hair cells and restore hearing function, but mechanisms controlling regeneration are not understood. The secreted protein bone morphogenetic protein 4 (BMP4) regulates inner ear morphogenesis and hair cell development. To investigate mechanisms controlling hair cell regeneration in birds, we examined expression and function of BMP4 in the auditory epithelia (basilar papillae) of chickens of either sex after hair cell destruction by ototoxic antibiotics...
May 2, 2018: Hearing Research
https://www.readbyqxmd.com/read/29753589/development-of-a-highly-sensitive-pcr-dna-chip-method-to-detect-mycoplasmas-in-a-veterinary-modified-live-vaccine
#19
Sylvie Mbelo, Virginie Gay, Stephanie Blanchard, Eric Abachin, Stephanie Falque, Jacques Lechenet, Hervé Poulet, Blandine de Saint-Vis
Mycoplasmas are potential contaminants that introduce undesirable changes in mammalian cell cultures. They frequently contaminate cell substrates and other starting materials used for manufacturing cell-derived biologics, such as vaccines and pharmaceutical products. Mycoplasma purity testing of live vaccines, active ingredients, raw material, and seed lots is required during vaccine production. Previously, testing using a time-consuming, costly 28-day culture assay, which lacks sensitivity for species that do not grow in culture, was required in the European Pharmacopoeia (Ph...
May 9, 2018: Biologicals: Journal of the International Association of Biological Standardization
https://www.readbyqxmd.com/read/29743260/cellular-nucleic-acid-binding-protein-is-vital-to-testis-development-and-spermatogenesis-in-mice
#20
Bo Zheng, Tingting Gao, Jun Yu, Yueshuai Guo, Cong Shen, Xi Zhang, Hong Li, Xiaoyan Huang
The cellular nucleic acid-binding protein (CNBP), also known as zinc finger protein 9 (ZNF9), is a highly conserved zinc finger protein that is strikingly conserved among vertebrates. Data collected from low vertebrates showed that CNBP is expressed at high levels and distributed in the testes during spermatogenesis. However, the location and function of CNBP in mammalian testes is not well known. Here, by neonatal mouse testis culture and SSC culture methods, we studied the effect of CNBP knockdown on neonatal testicular development...
May 9, 2018: Reproduction: the Official Journal of the Society for the Study of Fertility
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