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https://www.readbyqxmd.com/read/28549037/evaluation-of-a-dna-extraction-and-purification-protocol-using-archived-formalin-fixed-paraffin-embedded-tissues-for-braf-mutations-analysis-in-papillary-thyroid-microcarcinomas
#1
Adela Nechifor-Boilă, Andrada Loghin, Françoise Descotes, Myriam Decaussin-Petrucci, Angela Borda
The isolation of good quality genomic DNA from formalin-fixed, paraffin-embedded tissues is challenging, especially in cases of small tissue specimens. The aim of our study was to evaluate a DNA extraction protocol using formalin-fixed, paraffin-embedded tissues in our laboratory and apply this method to a series of papillary thyroid microcarcinomas (PTMCs). A total of 25 PTMCs and 3 papillary thyroid carcinoma control cases were included in the study. We assessed a DNA extraction protocol on the basis of a precipitation method (MasterPure DNA purification kit, Epicentre), according to the manufacturer's instructions...
May 25, 2017: Applied Immunohistochemistry & Molecular Morphology: AIMM
https://www.readbyqxmd.com/read/28548037/moonlighting-glycolytic-enzyme-glyceraldehyde-3-phosphate-dehydrogenase-gapdh-is-required-for-efficient-hepatitis-c-virus-and-dengue-virus-infections-in-human-huh-7-5-1-cells
#2
Meera Raj, Mary Langley, Steven J McArthur, François Jean
Hijacking of cellular biosynthetic pathways by human enveloped viruses is a shared molecular event essential for the viral lifecycle. In this study, the accumulating evidence of the importance of human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in the host secretory pathway led us to hypothesize that this moonlighting enzyme could play a key role in the lifecycle steps of two important Flaviviridae members, hepatitis C virus (HCV) and dengue virus (DENV). We used short interfering RNA (siRNA)-mediated knockdown of human GAPDH in Huh-7...
May 26, 2017: Journal of General Virology
https://www.readbyqxmd.com/read/28546127/new-genes-for-accurate-normalization-of-qrt-pcr-results-in-study-of-ips-and-ips-derived-cells
#3
A S Artyukhov, E B Dashinimaev, V O Tsvetkov, A P Bolshakov, E V Konovalova, S N Kolbaev, E A Vorotelyak, A V Vasiliev
iPSC-derived cells (from induced pluripotent stem cells) are a useful source that provide a powerful and widely accepted tool for the study of various types of human cells in vitro. Indeed, iPSC-derived cells from patients with hereditary diseases have been shown to reproduce the hallmarks of these diseases in vitro, phenotypes that can then also be manipulated in vitro. Quantitative reverse transcription PCR (qRT-PCR) is often used to characterize the progress of iPSC differentiation, validate mature cell types and to determine levels of pathological markers...
May 22, 2017: Gene
https://www.readbyqxmd.com/read/28544358/the-expression-pattern-of-bcl11a-mdm2-and-pten-genes-in-b-cell-acute-lymphoblastic-leukemia
#4
Ling Xu, Hong Wu, Xiuli Wu, Yangqiu Li, Dongmei He
AIM: Bcl11a is closely associated with B-cell lymphoma and B-cell chronic lymphocytic leukemia. However, little is known about the expression character of Bcl11a gene in B-cell acute lymphoblastic leukemia (B-ALL). In previous study, our results showed there was a clear dysregulation in the global gene expression of the Bcl11a-suppressed B lymphoma cells. The aim of the study is to further evaluate the role of Bcl11a and the expression level of its related genes Mdm2, Pten in B-ALL patients...
May 24, 2017: Asia-Pacific Journal of Clinical Oncology
https://www.readbyqxmd.com/read/28536183/the-nlrp3-inflammasome-contributes-to-sarcopenia-and-lower-muscle-glycolytic-potential-in-old-mice
#5
Marin Jane McBride, Kevin P Foley, Donna M D'souza, Yujin E Li, Trevor C Lau, Thomas J Hawke, Jonathan D Schertzer
The mechanisms underpinning decreased skeletal muscle strength and slowing of movement during aging are ill-defined. "Inflammaging" - increased inflammation with advancing age - may contribute to aspects of sarcopenia, but little is known about the participatory immune components. We discovered that aging was associated with increased caspase-1 activity in mouse skeletal muscle. We hypothesized that the caspase-1 containing NLRP3 inflammasome contributes to sarcopenia in mice. Male C57BL/6J wild type (WT) and NLRP3(-/-) mice were aged to 10 months (adult) and 24 months (old)...
May 23, 2017: American Journal of Physiology. Endocrinology and Metabolism
https://www.readbyqxmd.com/read/28533178/mucin-and-carbohydrate-stimulated-adhesion-and-subproteome-changes-of-the-probiotic-bacterium-lactobacillus-acidophilus-ncfm
#6
Hasan Ufuk Celebioglu, Sita Vaag Olesen, Kennie Prehn, Sampo J Lahtinen, Susanne Brix, Maher Abou Hachem, Birte Svensson
Adhesion to intestinal mucosa is a crucial property for probiotic bacteria. Adhesion is thought to increase host-bacterial interactions, thus potentially enabling health benefits to the host. Molecular events connected with adhesion and surface proteome changes were investigated for the probiotic Lactobacillus acidophilus NCFM cultured with established or emerging prebiotic carbohydrates as carbon source and in the presence of mucin, the glycoprotein of the epithelial mucus layer. Variation in adhesion to HT29-cells and mucin was associated with carbon source and mucin-induced subproteome abundancy differences...
May 19, 2017: Journal of Proteomics
https://www.readbyqxmd.com/read/28523004/selection-of-suitable-reference-genes-for-quantitative-real-time-pcr-in-sapium-sebiferum
#7
Xue Chen, Yingji Mao, Shengwei Huang, Jun Ni, Weili Lu, Jinyan Hou, Yuting Wang, Weiwei Zhao, Minghao Li, Qiaojian Wang, Lifang Wu
Chinese tallow (Sapium sebiferum L.) is a promising landscape and bioenergy plant. Measuring gene expression by quantitative real-time polymerase chain reaction (qRT-PCR) can provide valuable information on gene function. Stably expressed reference genes for normalization are a prerequisite for ensuring the accuracy of the target gene expression level among different samples. However, the reference genes in Chinese tallow have not been systematically validated. In this study, 12 candidate reference genes (18S, GAPDH, UBQ, RPS15, SAND, TIP41, 60S, ACT7, PDF2, APT, TBP, and TUB) were investigated with qRT-PCR in 18 samples, including those from different tissues, from plants treated with sucrose and cold stresses...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28522607/nleb-ssek-effectors-from-citrobacter-rodentium-escherichia-coli-and-salmonella-enterica-display-distinct-differences-in-host-substrate-specificity
#8
Samir El Qaidi, Kangming Chen, Adnan Halim, Lina Siukstaite, Christian Rueter, Ramon Hurtado-Guerrero, Henrik Clausen, Philip R Hardwidge
Many Gram-negative bacterial pathogens use a syringe-like apparatus called a type III secretion system to inject virulence factors into host cells. Some of these effectors are enzymes that modify host proteins to subvert their normal functions. NleB is a glycosyltransferase that modifies host proteins with N-acetyl-D-glucosamine to inhibit antibacterial and inflammatory host responses. NleB is conserved among the attaching/effacing pathogens enterohemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC), and Citrobacter rodentium...
May 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28515175/role-for-reactive-oxygen-species-in-flow-stimulated-inner-medullary-collecting-duct-endothelin-1-production
#9
Will Wheatley, Donald E Kohan
Inner medullary collecting duct (IMCD)-derived endothelin-1 (ET-1) is stimulated by volume expansion, in part through augmented luminal flow, whereupon it can elicit natriuresis and diuresis. Since flow can alter nitric oxide (NO) and reactive oxygen species (ROS), both of which can affect collecting duct salt transport, we asked whether NO and/or ROS mediate flow-stimulated IMCD ET-1. Mouse IMCD3 cells were exposed to flow and ET-1/GAPDH mRNA assessed. A shear stress of 10 dyne/cm2 for 1 hr increased ET-1 mRNA by 4-fold compared to no flow (ET-1 flow response)...
May 17, 2017: American Journal of Physiology. Renal Physiology
https://www.readbyqxmd.com/read/28513228/relationship-between-chop-gadd153-and-unstable-human-carotid-atherosclerotic-plaque
#10
Suping Wang, Meiyan Zhang, Zanhua Liu, Wuyang Yang, Junwei Shi, Victor Dean, Dong Chen
BACKGROUND AND AIMS: The signaling protein C/EBP homologous protein (CHOP) and corresponding growth-arrest-and-DNA-damage-inducible gene 153 (GADD153) is associated with endoplasmic reticulum stress (ERS), which can lead to apoptosis. Our study aims to elucidate the role of CHOP/GADD153 in unstable atherosclerotic (AS) plaque formation isolated from confounding factors such as diabetes mellitus, primary hyperlipidemia, autoimmune deficiencies/abnormalities, essential hypertension, chronic heart failure, chronic kidney disease, and smoking...
May 17, 2017: British Journal of Neurosurgery
https://www.readbyqxmd.com/read/28508372/effect-of-second-hand-tobacco-smoke-on-the-nitration-of-brain-proteins-a-systems-biology-and-bioinformatics-approach
#11
Firas H Kobeissy, Joy Guingab-Cagmat, Adriaan W Bruijnzeel, Mark S Gold, Kevin Wang
Second-hand smoke (SHS) exposure leads to the death of approximately 48,000 nonsmokers per year in the United States alone. SHS exposure has been associated with cardiovascular, respiratory, and neurodegenerative diseases. While cardiac function abnormalities and lung cancer due to SHS have been well characterized, brain injury due to SHS has not undergone a full systematic evaluation. Oxidative stress and nitration have been associated with smoking and SHS exposure. Animal studies suggest that exposure to tobacco smoke increases oxidative stress...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28507026/lineage-specific-metabolic-properties-and-vulnerabilities-of-t-cells-in-the-demyelinating-central-nervous-system
#12
Scott M Seki, Max Stevenson, Abagail M Rosen, Sanja Arandjelovic, Lelisa Gemta, Timothy N J Bullock, Alban Gaultier
Multiple sclerosis (MS) is a disease that is characterized by immune-mediated destruction of CNS myelin. Current MS therapies aim to block peripheral immune cells from entering the CNS. Although these treatments limit new inflammatory activity in the CNS, no treatment effectively prevents long-term disease progression and disability accumulation in MS patients. One explanation for this paradox is that current therapies are ineffective at targeting immune responses already present in the CNS. To this end, we sought to understand the metabolic properties of T cells that mediate ongoing inflammation in the demyelinating CNS...
May 15, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28495560/characterization-of-two-cryptic-plasmids-from-kocuria-palustris-ipufs-1-and-construction-of-novel-escherichia-coli-kocuria-shuttle-vector-for-biocatalysis
#13
Hiroshi Toda, Takashi Koyanagi, Toshiki Enomoto, Nobuya Itoh
Two cryptic plasmids, designated pKPAL1 and pKPAL2, were identified from the gram-positive bacterium Kocuria palustris IPUFS-1, which was isolated from a fish source. The 2251-bp and 2488-bp circular genomes of pKPAL1 and pKPAL2, respectively, were sequenced. Subsequent open reading frame (ORF) and homology search analyses suggested that pKPAL1 and pKPAL2 possess two and three ORFs, respectively, and encode the putative replication proteins, RepA and RepB, like the genomes of several plasmids in gram-positive bacteria...
May 8, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28493611/fucoidan-ameliorates-pancreatic-%C3%AE-cell-death-and-impaired-insulin-synthesis-in-streptozotocin-treated-%C3%AE-cells-and-mice-via-a-sirt-1-dependent-manner
#14
Wen-Chun Yu, Yen-Lin Chen, Pai-An Hwang, Tso-Hsiao Chen, Tz-Chong Chou
SCOPE: Several beneficial biological functions of fucoidan (FO) isolated from brown algae have been demonstrated. The purpose of this study was to investigate whether FO derived from Sargassum hemiphyllum ameliorates pancreatic β-cell damage and impaired insulin synthesis under diabetic condition. METHODS AND RESULTS: The effects of FO were studied in streptozotocin (STZ)-treated pancreatic β-cell line, NIT-1cells, and mice. The cell apoptosis, protein analyses, histological examination, and pancreatic function assays were performed...
May 11, 2017: Molecular Nutrition & Food Research
https://www.readbyqxmd.com/read/28490769/increased-cerebral-expressions-of-mmps-cldn5-ocln-zo1-and-aqps-are-associated-with-brain-edema-following-fatal-heat-stroke
#15
Yu Du, Jing-Tao Xu, Hong-Nian Jin, Rui Zhao, Dong Zhao, Si-Hao Du, Ye Xue, Xiao-Li Xie, Qi Wang
Human brain samples were collected from 46 autopsy cases, including 23 fatal heat stroke cases and 23 age-matched controls. Nine candidate reference genes (PES1, POLR2A, IPO8, HMBS, SDHA, GAPDH, UBC, B2M, ACTB) were evaluated in the cerebral cortex of 10 forensic autopsy cases (5 heat stroke and 5 controls), using the geNorm module in qBase(plus) software. SDHA, POLR2A, IPO8 and HMBS were identified as the most stable reference genes. Using these validated reference genes, mRNA expressions of Matrix metalloproteinases (MMPs, MMP2 and MMP9), Claudin5 (CLDN5), Occludin (OCLN), Zona occludens protein-1 (ZO1) and Aquaporins (AQPs, AQP1 and AQP4) in the cerebral cortex were examined...
May 10, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28488596/the-impact-of-microfluidic-mixing-of-triblock-micelleplexes-on-in-vitro-formula-see-text-in-vivo-gene-silencing-and-intracellular-trafficking
#16
Daniel P Feldmann, Yuran Xie, Steven K Jones, Dongyue Yu, Anna Moszczynska, Olivia M Merkel
The triblock copolymer polyethylenimine-polycaprolactone-polyethylene glycol (PEI-PCL-PEG) has been shown to spontaneously assemble into nano-sized particulate carriers capable of complexing with nucleic acids for gene delivery. The objective of this study was to investigate micelleplex characteristics, their in vitro and in vivo fate following microfluidic preparation of siRNA nanoparticles compared to the routinely used batch reactor mixing technique. Herein, PEI-PCL-PEG nanoparticles were prepared with batch reactor or microfluidic mixing techniques and characterized by various biochemical assays and in cell culture...
June 2, 2017: Nanotechnology
https://www.readbyqxmd.com/read/28479730/anti-hepatocarcinogenic-and-anti-oxidant-effects-of-mangrove-plant-scyphiphora-hydrophyllacea
#17
Sameera R Samarakoon, Chanthirika Shanmuganathan, Meran K Ediriweera, Poorna Piyathilaka, Kamani H Tennekoon, Ira Thabrew, Prasanna Galhena, E Dilip De Silva
CONTEXT: Scyphiphora hydrophyllacea is a shrub mangrove plant of the family Rubiaceae and not yet been studied for anti-hepatocarcinogenic effects. OBJECTIVES: We investigated possible in vitro anti-hepatocarcinogenic and antioxidant properties of S. hydrophyllacea. MATERIALS AND METHODS: Dried leaves of S. hydrophyllacea were sequentially extracted into hexane, chloroform, ethyl acetate, and methanol and tested for cytotoxicity on HepG2 cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and sulforhodamine B assays, and for antioxidant activities by the free radical 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) assays...
January 2017: Pharmacognosy Magazine
https://www.readbyqxmd.com/read/28472872/herv-k-and-w-expression-in-peripheral-mononuclear-cells-of-children-with-henoch-sch%C3%A3-nlein-purpura-and-relation-with-tlrs-activation
#18
Massimiliano Bergallo, Elisa Loiacono, Ilaria Galliano, Paola Montanari, Licia Peruzzi, Pier-Angelo Tovo, Rosanna Coppo
BACKGROUND: HERVs expression seems impaired in several diseases, ranging from autoimmune to neoplastic disorders. However, various stimuli may re-activate HERVs transcription. Henoch-Schönlein purpura is the most common cause of vasculitis in children. The role of microbial antigens in the pathogenesis of HSP remains elusive. TLRs are the first line of defense for the host to initiate an immune and inflammatory response. We aimed to investigate HERV, K and W expression in peripheral mononuclear cells of children with HSP and relation with TLRs activation...
May 4, 2017: Minerva Pediatrica
https://www.readbyqxmd.com/read/28453515/reliable-reference-genes-for-normalization-of-gene-expression-data-in-tea-plants-camellia-sinensis-exposed-to-metal-stresses
#19
Ming-Le Wang, Qing-Hui Li, Hua-Hong Xin, Xuan Chen, Xu-Jun Zhu, Xing-Hui Li
Tea plants [Camellia sinensis (L.) O. Kuntze] are an important leaf-type crop that are widely used for the production of non-alcoholic beverages in the world. Exposure to excessive amounts of heavy metals adversely affects the quality and yield of tea leaves. To analyze the molecular responses of tea plants to heavy metals, a reliable quantification of gene expression is important and of major importance herein is the normalization of the measured expression levels for the target genes. Ideally, stably expressed reference genes should be evaluated in all experimental systems...
2017: PloS One
https://www.readbyqxmd.com/read/28451145/competitive-method-based-electrochemiluminescent-assay-with-protein-nucleotide-conversion-for-ratio-detection-to-efficiently-monitor-the-drug-resistance-of-cancer-cells
#20
Wen-Bin Liang, Ming-Zhen Yang, Ying Zhuo, Ying-Ning Zheng, Cheng-Yi Xiong, Ya-Qin Chai, Ruo Yuan
A simple and highly-efficient approach to monitor the expression of P-glycoprotein (P-gp) in cells was urgently needed to demonstrate the drug resistance of cancer cells. Herein, a competitive method-based electrochemiluminescent (ECL) assay with a single ECL indicator was proposed for the first time to efficiently estimate the concentration ratio of two proteins. By converting the different proteins to partially coincident nucleotide sequences via a sandwich type immunoassay on magnetic beads, the concentration ratio related ECL signals could be obtained via competitive nucleotide hybridization on an electrode surface...
December 1, 2016: Chemical Science
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