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https://www.readbyqxmd.com/read/29140568/crispr-cas9-based-safe-harbor-gene-editing-in-rhesus-ipscs
#1
Ravi Chandra Yada, John W Ostrominski, Ilker Tunc, So Gun Hong, Jizhong Zou, Cynthia E Dunbar
NHP iPSCs provide a unique opportunity to test safety and efficacy of iPSC-derived therapies in clinically relevant NHP models. To monitor these cells in vivo, there is a need for safe and efficient labeling methods. Gene insertion into genomic safe harbors (GSHs) supports reliable transgene expression while minimizing the risk the modification poses to the host genome or target cell. Specifically, this protocol demonstrates targeting of the adeno-associated virus site 1 (AAVS1), one of the most widely used GSH loci in the human genome, with CRISPR/Cas9, allowing targeted marker or therapeutic gene insertion in rhesus macaque induced pluripotent stem cells (RhiPSCs)...
November 15, 2017: Current Protocols in Stem Cell Biology
https://www.readbyqxmd.com/read/29139018/vaccination-against-hydatidosis-molecular-cloning-and-optimal-expression-of-the-eg95nc-recombinant-antigen-in-escherichia-coli
#2
M Jazouli, M Lightowlers, C G Gauci, K Tadlaoui, A Belmlih, M M Ennaji, M Elharrak
Cystic echinococcosis (CE) is a widely distributed zoonosis that is highly endemic in the Mediterranean basin. The disease represents a serious public health threat and causes economic losses. The parasite life-cycle involves dogs and ruminants as definitive and intermediate hosts; humans are accidently infected, causing serious clinical issues. Vaccination of ruminants and dog treatments represent the most efficient measures to prevent parasite transmission. The recombinant protein vaccine, EG95, has been used successfully in sheep vaccine trials against CE in several countries...
November 14, 2017: Protein Journal
https://www.readbyqxmd.com/read/29138301/genomic-variation-and-evolution-of-vibrio-parahaemolyticus-st36-over-the-course-of-a-transcontinental-epidemic-expansion
#3
Jaime Martinez-Urtaza, Ronny van Aerle, Michel Abanto, Julie Haendiges, Robert A Myers, Joaquin Trinanes, Craig Baker-Austin, Narjol Gonzalez-Escalona
Vibrio parahaemolyticus is the leading cause of seafood-related infections with illnesses undergoing a geographic expansion. In this process of expansion, the most fundamental change has been the transition from infections caused by local strains to the surge of pandemic clonal types. Pandemic clone sequence type 3 (ST3) was the only example of transcontinental spreading until 2012, when ST36 was detected outside the region where it is endemic in the U.S. Pacific Northwest causing infections along the U.S. northeast coast and Spain...
November 14, 2017: MBio
https://www.readbyqxmd.com/read/29137976/proteomic-analysis-of-cho-cell-lines-producing-high-and-low-quantities-of-a-recombinant-antibody-before-and-after-selection-with-methotrexate
#4
Ruth Hausmann, Ivana Chudobová, Holger Spiegel, Stefan Schillberg
High levels of recombinant protein production in Chinese hamster ovary (CHO) cells can be achieved by amplification of transgenes using the dihydrofolate reductase/methotrexate (DHFR/MTX) system. With the aim to identify predictive markers enabling the preselection of suitable high producing clones we investigated the impact of MTX-based gene amplification on two CHO cells lines producing different levels of a human monoclonal antibody by carrying out a comparative proteome analysis. The difference in antibody yield between the high and low producer was 15-fold before and 245-fold after MTX selection...
November 11, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/29135534/the-multiple-faces-of-programmed-cell-death-ligand-1-expression-in-malignant-and-nonmalignant-cells
#5
Edwin R Parra, Pamela Villalobos, Jaime Rodriguez-Canales
Preliminary data suggest that tumor expression of programmed cell death ligand 1 (PD-L1) protein in human cancers, as determined by immunohistochemistry in formalin-fixed, paraffin-embedded tissue samples, may predict clinical response to anti-PD-1/PD-L1 therapy. PD-L1 is not a specific tumor marker and its expression is also observed in various nonmalignant cells, such as macrophages and lymphocytes, causing confusion in immunohistochemistry analysis when these inflammatory cells are overlapping with tumors cells...
November 13, 2017: Applied Immunohistochemistry & Molecular Morphology: AIMM
https://www.readbyqxmd.com/read/29133557/a-novel-mechanism-of-inactivating-antibacterial-nitro-compounds-in-the-human-pathogen-staphylococcus-aureus-by-overexpression-of-a-nadh-dependent-flavin-nitroreductase
#6
Ebaa M El-Hossary, Konrad U Förstner, Patrice François, Damien Baud, Karin Streker, Jacques Schrenzel, Knut Ohlsen, Ulrike Holzgrabe
Recently, the nitro substituted bisquaternary bisnaphthalimides were reported to have substantial anti-infective activity against Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). Here, we selected resistant S. aureus clones by cultivation in increasing concentrations of the most active compound MT02. Interestingly, MT02 resistant variants induced a diffusible red color of the broth. Chromatographic and spectroscopic investigations revealed a stepwise reduction of the bisquaternary bisnaphthalimides̀ nitro groups to amino groups...
November 13, 2017: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/29133551/copper-ions-and-coordination-complexes-as-novel-carbapenem-adjuvants
#7
Karrera Y Djoko, Maud E S Achard, Minh-Duy Phan, Alvin W Lo, Manfredi Miraula, Sasiprapa Prombhul, Steven J Hancock, Kate M Peters, Hanna Sidjabat, Patrick N Harris, Nataša Mitić, Timothy R Walsh, Gregory J Anderson, William M Shafer, David L Paterson, Gerhard Schenk, Alastair G McEwan, Mark A Schembri
Carbapenem-resistant Enterobacteriaceae are an urgent threat to global human health. These organisms produce β-lactamases with carbapenemase activity, such as the metallo-β-lactamase NDM-1, which is notable due to its association with mobile genetic elements and the lack of a clinically useful inhibitor. Here we examined the ability of copper to inhibit the activity of NDM-1 and explored the potential of a copper coordination complex as a mechanism to efficiently deliver copper as an adjuvant in clinical therapeutics...
November 13, 2017: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/29133173/effects-of-norquetiapine-the-active-metabolite-of-quetiapine-on-cloned-herg-potassium-channels
#8
Hong Joon Lee, Jin-Sung Choi, Bok Hee Choi, Sang June Hahn
Quetiapine is an atypical antipsychotic drug that is widely used for the treatment of schizophrenia. It is mainly metabolized by a cytochrome P450 system in the liver. Norquetiapine is a major active metabolite in humans with a pharmacological profile that differs distinctly from that of quetiapine. We used the whole-cell patch-clamp technique to investigate the effects of norquetiapine on hERG channels that are stably expressed in HEK cells. Quetiapine and norquetiapine inhibited the hERG tail currents at -50mV in a concentration-dependent manner with IC50 values of 8...
November 10, 2017: Neuroscience Letters
https://www.readbyqxmd.com/read/29132372/hepatitis-b-virus-inhibits-the-in-vivo-and-in-vitro-synthesis-and-secretion-of-apolipoprotein-c3
#9
Chengliang Zhu, Hengcheng Zhu, Hui Song, Limin Xu, Longxuan Li, Fang Liu, Xinghui Liu
BACKGROUND: Hepatitis B virus (HBV) infection in the body can damage liver cells and cause disorders in blood lipid metabolism. Apolipoprotein C3 (ApoC3) plays an important role in the regulation of lipid metabolism, but no study on the HBV regulation of ApoC3 has been reported. This purpose of this study was to investigate the effect of HBV on ApoC3 expression and its regulatory mechanism. METHODS: The expression levels of ApoC3 mRNA and protein in the human hepatoma cell lines HepG2 and HepG2...
November 13, 2017: Lipids in Health and Disease
https://www.readbyqxmd.com/read/29130672/-effects-of-cellular-density-on-the-induction-of-suspension-globe-of-ovarian-cancer-stem-cells
#10
Mian-Zhi Chen, Lan Zhong, Dong-Mei Wei, Xiao-Yu Niu
OBJECTIVE: To determine the effect of cellular density on the separation and identification of cancer stem cells from human ovarian clear cell carcinoma cell line ES-2 and adenocarcinoma cell line A2780. METHODS: ES-2 and A2780 cells were cultured with human recombinant epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) and bovine serum albumin and insulin in serum free medium. The cancer stem cells were obtained through serial passages. Changes in cell morphology,expressions of surface marker CD133 and CD44,and soft AGAR clone forming in the stem cells were examined under different cell density,either in serum-supplemented medium (SSM group) or in serum free medium (SFM group)...
September 2017: Sichuan da Xue Xue Bao. Yi Xue Ban, Journal of Sichuan University. Medical Science Edition
https://www.readbyqxmd.com/read/29129319/modeling-the-interplay-between-neurons-and-astrocytes-in-autism-using-human-induced-pluripotent-stem-cells
#11
Fabiele Baldino Russo, Beatriz Camille Freitas, Graciela Conceição Pignatari, Isabella Rodrigues Fernandes, Jonathan Sebat, Alysson Renato Muotri, Patricia Cristina Baleeiro Beltrão-Braga
BACKGROUND: Autism spectrum disorder (ASD) is a neurodevelopmental disorder with unclear etiology and imprecise genetic causes. The main goal of this work was to investigate neuronal connectivity and the interplay between neurons and astrocytes from individuals with nonsyndromic ASD using induced pluripotent stem cells. METHODS: Induced pluripotent stem cells were derived from a clinically well-characterized cohort of three individuals with nonsyndromic ASD sharing common behaviors and three control subjects, two clones each...
October 3, 2017: Biological Psychiatry
https://www.readbyqxmd.com/read/29128907/a-new-conditionally-immortalized-human-fetal-brain-pericyte-cell-line-establishment-and-functional-characterization-as-a-promising-tool-for-human-brain-pericyte-studies
#12
Kenta Umehara, Yuchen Sun, Satoshi Hiura, Koki Hamada, Motoyuki Itoh, Keita Kitamura, Motohiko Oshima, Atsushi Iwama, Kosuke Saito, Naohiko Anzai, Kan Chiba, Hidetaka Akita, Tomomi Furihata
While pericytes wrap around microvascular endothelial cells throughout the human body, their highest coverage rate is found in the brain. Brain pericytes actively contribute to various brain functions, including the development and stabilization of the blood-brain barrier (BBB), tissue regeneration, and brain inflammation. Accordingly, detailed characterization of the functional nature of brain pericytes is important for understanding the mechanistic basis of brain physiology and pathophysiology. Herein, we report on the development of a new human brain pericyte cell line, hereafter referred to as the human brain pericyte/conditionally immortalized clone 37 (HBPC/ci37)...
November 11, 2017: Molecular Neurobiology
https://www.readbyqxmd.com/read/29128656/molecular-and-functional-characterization-of-two-malic-enzymes-from-leishmania-parasites
#13
Lucila Giordana, Máximo Hernán Sosa, Alejandro E Leroux, Elkin F Rodas Mendoza, Patricia Petray, Cristina Nowicki
Leishmania parasites cause a broad spectrum of clinical manifestations in humans and the available clinical treatments are far from satisfactory. Since these pathogens require large amounts of NADPH to maintain intracellular redox homeostasis, oxidoreductases that catalyze the production of NADPH are considered as potential drug targets against these diseases. In the sequenced genomes of most Leishmania spp. two putative malic enzymes (MEs) with an identity of about 55% have been identified. In this work, the ME from L...
November 8, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29128358/plant-salt-tolerance-mechanism-a-review
#14
Wenji Liang, Xiaoli Ma, Peng Wan, Lianyin Liu
Almost all crops that are important to humans are sensitive to high salt concentration in the soil. The presence of salt in soil is one of the most significant abiotic stresses in farming. Therefore, improving plant salt tolerance and increasing the yield and quality of crops in salty land is vital. Transgenic technology is a fast and effective method to obtain salt-tolerant varieties. At present, many scholars have studied salt damage to plant and plant salt-tolerance mechanism. These scholars have cloned a number of salt-related genes and achieved high salt tolerance for transgenic plants, thereby showing attractive prospects...
November 8, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29126422/changes-in-total-and-differential-leukocyte-counts-during-the-clinically-silent-liver-phase-in-a-controlled-human-malaria-infection-in-malaria-na%C3%A3-ve-dutch-volunteers
#15
Marlies E van Wolfswinkel, Marijke C C Langenberg, Linda J Wammes, Robert W Sauerwein, Rob Koelewijn, Cornelus C Hermsen, Jaap J van Hellemond, Perry J van Genderen
BACKGROUND: Both in endemic countries and in imported malaria, changes in total and differential leukocyte count during Plasmodium falciparum infection have been described. To study the exact dynamics of differential leukocyte counts and their ratios, they were monitored in a group of healthy non-immune volunteers in two separate Controlled Human Malaria Infection (CHMI) studies. METHODS: In two CHMI trials, CHMI-a and CHMI-b, 15 and 24 healthy malaria-naïve volunteers, respectively, were exposed to bites of infected mosquitoes, using the P...
November 10, 2017: Malaria Journal
https://www.readbyqxmd.com/read/29124072/decitabine-induced-changes-in-human-myelodysplastic-syndrome-cell-line-skm-1-are-mediated-by-foxo3a-activation
#16
Wen Zeng, Hanjun Dai, Ming Yan, Xiaojun Cai, Hong Luo, Min Ke, Zeming Liu
The epigenetic silencing of tumor suppressor genes in myelodysplastic syndromes (MDS) can potentially confer a growth advantage to individual cellular clones. Currently, the recommended treatment for patients with high-risk MDS is the methylation agent decitabine (DAC), a drug that can induce the reexpression of silenced tumor suppressor genes. We investigated the effects of DAC treatment on the myeloid MDS cell line SKM-1 and investigated the role of FOXO3A, a potentially tumor-suppressive transcription factor, by silencing its expression prior to DAC treatment...
2017: Journal of Immunology Research
https://www.readbyqxmd.com/read/29123516/avidity-and-bystander-suppressive-capacity-of-human-regulatory-t-cells-expressing-de-novo-autoreactive-t-cell-receptors-in-type-1-diabetes
#17
Wen-I Yeh, Howard R Seay, Brittney Newby, Amanda L Posgai, Filipa Botelho Moniz, Aaron Michels, Clayton E Mathews, Jeffrey A Bluestone, Todd M Brusko
The ability to alter antigen specificity by T-cell receptor (TCR) or chimeric antigen receptor (CAR) gene transfer has facilitated personalized cellular immune therapies in cancer. Inversely, this approach can be harnessed in autoimmune settings to attenuate inflammation by redirecting the specificity of regulatory T cells (Tregs). Herein, we demonstrate efficient protocols for lentiviral gene transfer of TCRs that recognize type 1 diabetes-related autoantigens with the goal of tissue-targeted induction of antigen-specific tolerance to halt β-cell destruction...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/29118372/human-single-chain-transbodies-that-bound-to-domain-i-of-non-structural-protein-5a-ns5a-of-hepatitis-c-virus
#18
Kittirat Glab-Ampai, Monrat Chulanetra, Aijaz Ahmad Malik, Thanate Juntadech, Jeeraphong Thanongsaksrikul, Potjanee Srimanote, Kanyarat Thueng-In, Nitat Sookrung, Pongsri Tongtawe, Wanpen Chaicumpa
A safe and broadly effective direct acting anti-hepatitis C virus (HCV) agent that can withstand the viral mutation is needed. In this study, human single chain antibody variable fragments (HuscFvs) to conserved non-structural protein-5A (NS5A) of HCV were produced by phage display technology. Recombinant NS5A was used as bait for fishing-out the protein bound-phages from the HuscFv-phage display library. NS5A-bound HuscFvs produced by five phage transfected-E. coli clones were linked molecularly to nonaarginine (R9) for making them cell penetrable (become transbodies)...
November 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29117786/molecular-characterization-and-functional-analysis-of-pr-1-like-proteins-identified-from-the-wheat-head-blight-fungus-fusarium-graminearum
#19
Shunwen Lu, Michael Edwards
The group 1 pathogenesis-related (PR-1) proteins originally identified from plants and their homologues are also found in other eukaryotic kingdoms. Studies on non-plant PR-1-like (PR-1L) proteins have been pursued widely in humans/animals but rarely in filamentous ascomycetes. Here we report the characterization of four PR-1L proteins identified from the ascomycete fungus Fusarium graminearum, the primary cause of Fusarium head blight of wheat and barley. Molecular cloning revealed that the four FgPR-1L proteins are all encoded by small open reading frames (612 to 909 bp) that are often interrupted by introns, in contrast to plant PR-1 genes that lack introns...
November 8, 2017: Phytopathology
https://www.readbyqxmd.com/read/29117182/structural-and-functional-analysis-of-coral-hypoxia-inducible-factor
#20
Didier Zoccola, Jonas Morain, Gilles Pagès, Natacha Caminiti-Segonds, Sandy Giuliano, Sylvie Tambutté, Denis Allemand
Tissues of symbiotic Cnidarians are exposed to wide, rapid and daily variations of oxygen concentration. Indeed, during daytime, intracellular O2 concentration increases due to symbiont photosynthesis, while during night, respiration of both host cells and symbionts leads to intra-tissue hypoxia. The Hypoxia Inducible Factor 1 (HIF-1) is a heterodimeric transcription factor used for maintenance of oxygen homeostasis and adaptation to hypoxia. Here, we carried out a mechanistic study of the response to variations of O2 concentrations of the coral model Stylophora pistillata...
2017: PloS One
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