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https://www.readbyqxmd.com/read/27893852/a-comparative-study-of-smn-protein-and-mrna-in-blood-and-fibroblasts-in-patients-with-spinal-muscular-atrophy-and-healthy-controls
#1
Renske I Wadman, Marloes Stam, Marc D Jansen, Yana van der Weegen, Camiel A Wijngaarde, Oliver Harschnitz, Peter Sodaar, Kees P J Braun, Dennis Dooijes, Henny H Lemmink, Leonard H van den Berg, W Ludo van der Pol
BACKGROUND: Clinical trials to test safety and efficacy of drugs for patients with spinal muscular atrophy (SMA) are currently underway. Biomarkers that document treatment-induced effects are needed because disease progression in childhood forms of SMA is slow and clinical outcome measures may lack sensitivity to detect meaningful changes in motor function in the period of 1-2 years of follow-up during randomized clinical trials. OBJECTIVE: To determine and compare SMN protein and mRNA levels in two cell types (i...
2016: PloS One
https://www.readbyqxmd.com/read/27892794/effects-of-repeated-9-and-30-day-exposure-to-extremely-low-frequency-electromagnetic-fields-on-social-recognition-behavior-and-estrogen-receptors-expression-in-olfactory-bulb-of-wistar-female-rats
#2
C Bernal-Mondragón, V Arriaga-Avila, E Martínez-Abundis, B Barrera-Mera, O Mercado-Gómez, R Guevara-Guzmán
OBJECTIVE: We investigated the short- and long-term effects of extremely low-frequency electromagnetic fields (EMF) on social recognition behavior and expression of α- and β-estrogen receptors (ER). METHODS: Rats were exposed to 60-Hz electromagnetic fields for 9 or 30 days and tested for social recognition behavior. Immunohistochemistry and western blot assays were performed to evaluate α- and β-ER expression in the olfactory bulb of intact, ovariectomized (OVX), and ovariectomized+estradiol (E2) replacement (OVX+E2)...
November 28, 2016: Neurological Research
https://www.readbyqxmd.com/read/27889881/identification-of-reference-genes-and-mirnas-for-qrt-pcr-in-human-esophageal-squamous-cell-carcinoma
#3
Li Chen, Yulin Jin, Lin Wang, Fenghao Sun, Xiaodong Yang, Mengkun Shi, Cheng Zhan, Yu Shi, Qun Wang
It is important to select an appropriate reference gene and miRNA when using quantitative real-time polymerase chain reaction (qRT-PCR) to analyze gene and miRNA expression. However, many commonly used reference genes and miRNAs are not stably expressed and therefore not suitable for normalization or quantification of qRT-PCR data. This study aims to identify appropriate reference genes and miRNAs for use in human esophageal squamous carcinoma qRT-PCR analysis. Using data provided by The Cancer Genome Atlas, we identified DDX5, LAPTM4A, P4HB, RHOA, miR-28-5p, miR-34a-5p, and miR-186-5p as candidate reference genes and miRNAs...
January 2017: Medical Oncology
https://www.readbyqxmd.com/read/27879277/eukaryotic-translation-initiation-factor-5a2-promotes-metabolic-reprogramming-in-hepatocellular-carcinoma-cells
#4
Ting-Ting Cao, Shu-Hai Lin, Li Fu, Zhi Tang, Chi-Ming Che, Li-Yi Zhang, Xiao-Yan Ming, Teng-Fei Liu, Xu-Ming Tang, Bin-Bin Tan, Di Xiang, Feng Li, On-Yee Chan, Dan Xie, Zongwei Cai, Xin-Yuan Guan
Reprogramming of intracellular metabolism is common in liver cancer cells. Understanding the mechanisms of cell metabolic reprogramming may present a new basis for liver cancer treatment. In our previous study we reported that a novel oncogene eukaryotic translation initiation factor 5A2 (EIF5A2) promotes tumorigenesis under hypoxic condition. Here, we aim to investigate the role of EIF5A2 in cell metabolic reprogramming during hepatocellular carcinoma (HCC) development. In this study, we reported that the mRNA level of EIF5A2 was up-regulated in 59/105 (56...
November 22, 2016: Carcinogenesis
https://www.readbyqxmd.com/read/27878453/evaluation-of-stability-and-validation-of-reference-genes-for-rt-qpcr-expression-studies-in-rice-plants-under-water-deficit
#5
Priscila Ariane Auler, Letícia Carvalho Benitez, Marcelo Nogueira do Amaral, Isabel Lopes Vighi, Gabriela Dos Santos Rodrigues, Luciano Carlos da Maia, Eugenia Jacira Bolacel Braga
Many studies use strategies that allow for the identification of a large number of genes expressed in response to different stress conditions to which the plant is subjected throughout its cycle. In order to obtain accurate and reliable results in gene expression studies, it is necessary to use reference genes, which must have uniform expression in the majority of cells in the organism studied. RNA isolation of leaves and expression analysis in real-time quantitative polymerase chain reaction (RT-qPCR) were carried out...
November 23, 2016: Journal of Applied Genetics
https://www.readbyqxmd.com/read/27878251/glyceraldehyde-3-phosphate-dehydrogenase-promotes-cancer-growth-and-metastasis-through-upregulation-of-snail-expression
#6
Kaiyan Liu, Zhenjie Tang, Amin Huang, Ping Chen, Panpan Liu, Jing Yang, Wenhua Lu, Jianwei Liao, Yicheng Sun, Shijun Wen, Yumin Hu, Peng Huang
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) plays an important role in multiple cellular functions including metabolism and gene transcription. Our previous study showed that GAPDH expression was elevated in colon cancer and further upregulated in liver metastatic tissues, suggesting a possilbe role of GAPDH in promoting cancer metastasis. The present study was designed to investigate the underlying mechanism, using multiple experimental approaches including genetic silencing of GAPDH expression by short hairpin RNA (shRNA) and biochemcial/molecular analyses of the key events involved in glycolytic metabolism and epithelial-mesenchymal transition (EMT)...
November 18, 2016: International Journal of Oncology
https://www.readbyqxmd.com/read/27875551/crystal-structures-of-group-b-streptococcus-glyceraldehyde-3-phosphate-dehydrogenase-apo-form-binary-and-ternary-complexes
#7
Norbert Schormann, Chapelle A Ayres, Alexandra Fry, Todd J Green, Surajit Banerjee, Glen C Ulett, Debasish Chattopadhyay
Glyceraldehyde 3-phosphate dehydrogenase or GAPDH is an evolutionarily conserved glycolytic enzyme. It catalyzes the two step oxidative phosphorylation of D-glyceraldehyde 3-phosphate into 1,3-bisphosphoglycerate using inorganic phosphate and NAD+ as cofactor. GAPDH of Group B Streptococcus is a major virulence factor and a potential vaccine candidate. Moreover, since GAPDH activity is essential for bacterial growth it may serve as a possible drug target. Crystal structures of Group B Streptococcus GAPDH in the apo-form, two different binary complexes and the ternary complex are described here...
2016: PloS One
https://www.readbyqxmd.com/read/27871894/optimizing-gdna-extraction-from-fresh-frozen-meningioma-tissue-for-downstream-genetic-analysis
#8
D T Proctor, E H Yoo, Z Vujadinovic, S Lama, G van Marle, G R Sutherland
OBJECTIVE: Meningioma is the most common brain tumor. Genetic mutations in meningioma that include deletion of the neurofibromatosis type 2 gene, (NF2), offer diagnostic information on tumor behavior, recurrence and potential response to treatment. Obtaining high-grade genetic material is critical for accurate, sensitive and robust molecular testing. Currently, no standardized procedure exists for extracting gDNA from meningioma, and this problem was addressed in this report. METHOD: This study compared the yield and quality of extracted gDNA from patient meningioma specimens using an optimized phenol chloroform method and two commercial silica column-based extractions kits and tested respective performances as template in qPCR tests and multiplex ligation-dependent probe amplification (MLPA) NF2 screening...
November 19, 2016: Clinical Biochemistry
https://www.readbyqxmd.com/read/27869545/non-invasive-monitoring-of-3d-chondrogenic-constructs-using-molecular-beacon-nanosensors
#9
Li Min Tay, Christian Wiraja, David Yeo, Yingnan Wu, Zheng Yang, Yon Jin Chuah, Eng Hin Lee, Yuejun Kang, Chenjie Xu
Chondrogenic differentiation of human mesenchymal stem cells (MSCs) in 3D hydrogel holds promise as a method for repairing injured articular cartilage. Given MSC plasticity (its potential to mature into alternative lineages), non-destructive monitoring is critical for the optimization of chondrogenic differentiation conditions and the evaluation of the final product. However, conventional validation/assessment of the differentiation process (i.e. quantitative reverse transcription polymerase chain (qRT-PCR) and histology) are end-point assays requiring disruption of the sample...
November 21, 2016: Tissue Engineering. Part C, Methods
https://www.readbyqxmd.com/read/27867269/platyphylloside-isolated-from-betula-platyphylla-inhibit-adipocyte-differentiation-and-induce-lipolysis-via-regulating-adipokines-including-ppar%C3%AE-in-3t3-l1-cells
#10
Mina Lee, Sang Hyun Sung
BACKGROUND: Obesity causes or aggravates many health problems, both independently and in association with several pathological disorders, including Type II diabetes, hypertension, atherosclerosis, and cancer. Therefore, we screened small compounds isolated from natural products for the development of anti-obesity drugs. OBJECTIVE: The purpose of this study was to investigate the anti-adipogenic activities of platyphylloside, diarylheptanoid isolated from Betula platyphylla, which was selected based on the screening using 3T3-L1 cells...
October 2016: Pharmacognosy Magazine
https://www.readbyqxmd.com/read/27862669/association-between-nephrinuria-podocyturia-and-proteinuria-in-women-with-pre-eclampsia
#11
Itsuko Furuta, Tianyue Zhai, Satoshi Ishikawa, Takeshi Umazume, Kinuko Nakagawa, Takahiro Yamada, Mamoru Morikawa, Hisanori Minakami
AIM: Podocyte depletion in the kidney is associated with end-stage kidney disease (ESKD). Pre-eclampsia (PE) increases the risk of ESKD in later life. This study was performed to determine whether nephrinuria (soluble nephrin in the urine) is correlated with proteinuria and/or podocyturia (podocytes in the urine) in PE women. METHODS: Eighty-three urine samples, consisting of 45 and 38 samples from 27 normotensive and nine PE women, respectively, underwent simultaneous determination of nephrin, protein, and creatinine concentrations in the urine supernatant and quantitative analysis of podocyte-specific protein mRNA expression...
November 12, 2016: Journal of Obstetrics and Gynaecology Research
https://www.readbyqxmd.com/read/27862636/a-novel-method-for-a-high-enrichment-of-human-corneal-epithelial-stem-cells-for-genomic-analysis
#12
Jhansi Rani Kasinathan, Venkatesh Prajna Namperumalsamy, Muthukkaruppan Veerappan, Gowri Priya Chidambaranathan
Understanding the molecular mechanisms that regulate the corneal epithelial stem cells (CESCs) in maintaining corneal homeostasis remains elusive largely due to the lack of a specific marker for their isolation. This study aims to enrich CESCs from human donor limbal epithelium and to evaluate the level of enrichment based on expression of ΔNp63α, a putative CESC marker. A two-stage enrichment of CESCs was carried out. (a) The limbal basal epithelial cells were isolated by differential enzymatic treatment and five-fold enrichment was achieved from 2% of CESCs present in the total limbal epithelium...
December 2016: Microscopy Research and Technique
https://www.readbyqxmd.com/read/27859958/phylogeny-and-variability-of-colletotrichum-truncatum-associated-soybean-anthracnose-in-brazil
#13
Flávia Rogério, Maisa Ciampi-Guillardi, Marina Coan Goldoni Barbieri, Carlos Augusto Dórea Bragança, Claudine Dinali Santos Seixas, Álvaro Manuel Rodrigues Almeida, Nelson Sidnei Massola
AIMS: Fungal diseases are among the main factors limiting high yields of soybean crop. Colletotrichum isolates from soybean plants with anthracnose symptoms were studied from different regions and time periods in Brazil using molecular, morphological and pathogenic analyses. METHODS AND RESULTS: Bayesian phylogenetic inference of GAPDH, HIS3 and ITS-5.8S rDNA sequences, the morphologies of colony and conidia and inoculation tests on seeds and seedlings were performed...
November 8, 2016: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/27859598/reduction-of-gapdh-in-lenses-of-parkinson-s-disease-patients-a-possible-new-biomarker
#14
Alexa Klettner, Andreas Tholey, Alena Wiegandt, Elisabeth Richert, Bernhard Nölle, Günther Deuschl, Johann Roider, Susanne A Schneider
BACKGROUND: We previously showed that the protein pattern of lenses removed in cataract surgery differs between patients with Parkinson's disease and age-matched controls. In this study, we identified the protein reduced in abundance in the 34- to 37-kDa gel band. METHODS: During cataract surgery (phacoemulsification), we collected the rinsing fluid and lens particles from the eyes of PD patients and controls. Residual lens fragments in the supernatant of 3 PD patients and aged-matched controls were studied for protein profiles using liquid chromatography-mass spectrometry and Western blots...
November 10, 2016: Movement Disorders: Official Journal of the Movement Disorder Society
https://www.readbyqxmd.com/read/27835695/reference-genes-for-addressing-gene-expression-of-bladder-cancer-cell-models-under-hypoxia-a-step-towards-transcriptomic-studies
#15
Luís Lima, Cristiana Gaiteiro, Andreia Peixoto, Janine Soares, Manuel Neves, Lúcio Lara Santos, José Alexandre Ferreira
Highly aggressive, rapidly growing tumors contain significant areas of hypoxia or anoxia as a consequence of inadequate and/or irregular blood supply. During oxygen deprivation, tumor cells withstand a panoply of adaptive responses, including a shift towards anaerobic metabolism and the reprogramming of the transcriptome. One of the major mediators of the transcriptional hypoxic response is the hypoxia-inducible factor 1 (HIF-1), whose stabilization under hypoxia acts as an oncogenic stimulus contributing to chemotherapy resistance, invasion and metastasis...
2016: PloS One
https://www.readbyqxmd.com/read/27826623/-corrigendum-adenovirus-mediated-overexpression-of-bmp-9-inhibits-human-osteosarcoma-cell-growth-and-migration-through-downregulation-of-the-pi3k-akt-pathway
#16
Bo Li, Yuehua Yang, Shengdan Jiang, Binbin Ni, Ke Chen, Leisheng Jiang
Following the publication of this article, an interested reader drew to our attention an anomaly associated with the presentation of the western blots in Fig. 4B and C. These contained the same GAPDH control data, even though Fig. 4B and C were intended to show the inhibition of osteosarcoma cell migration by a recombinant adenovirus (ad) expressing bone morphogenetic protein 9 (adBMP-9) in two different cell lines, MG-63 and HOS, respectively. After having re-examined our original data, we realize that the same GAPDH control bands were inadvertently selected for Fig...
November 2016: International Journal of Oncology
https://www.readbyqxmd.com/read/27825774/identification-and-evaluation-of-reference-genes-for-expression-studies-by-rt-qpcr-during-embryonic-development-of-the-emerging-model-organism-macrobrachium-olfersii
#17
Michael L Jaramillo, Dib Ammar, Ruth L Quispe, Frank Guzman, Rogerio Margis, Evelise M Nazari, Yara M R Müller
RT-qPCR is a sensitive and highly efficient technique that is widely used in gene expression analysis and to provide insight into the molecular mechanisms underlying embryonic development. The freshwater prawn, Macrobrachium olfersii is an emerging model organism, but, the stable reference genes of this species need to be identified and validated for RT-qPCR analysis. Thus, the aim of this study was to evaluate the expression stability of six genes (β-act, GAPDH, EF-1α, RpL8, RpS6, AK) in embryos and in adult tissues (cerebral ganglia, muscle and hepatopancreas) of M...
November 5, 2016: Gene
https://www.readbyqxmd.com/read/27824922/identification-of-reference-genes-for-quantitative-real-time-pcr-in-date-palm-phoenix-dactylifera-l-subjected-to-drought-and-salinity
#18
Himanshu V Patankar, Dekoum V M Assaha, Rashid Al-Yahyai, Ramanjulu Sunkar, Mahmoud W Yaish
Date palm is an important crop plant in the arid and semi-arid regions supporting human population in the Middle East and North Africa. These areas have been largely affected by drought and salinity due to insufficient rainfall and improper irrigation practices. Date palm is a relatively salt- and drought-tolerant plant and more recently efforts have been directed to identifying genes and pathways that confer stress tolerance in this species. Quantitative real-time PCR (qPCR) is a promising technique for the analysis of stress-induced differential gene expression, which involves the use of stable reference genes for normalizing gene expression...
2016: PloS One
https://www.readbyqxmd.com/read/27824649/downregulation-of-microrna-21-in-colonic-cd3-t-cells-in-uc-remission
#19
Yugo Ando, Luca Mazzurana, Marianne Forkel, Kazuichi Okazaki, Mamiko Aoi, Peter T Schmidt, Jenny Mjösberg, Francesca Bresso
BACKGROUND: In active inflammatory bowel disease (IBD), microRNA expression profiling consistently features disease-specific signatures, and microRNA-21 (miR-21) has been shown to be upregulated in the inflamed colon of patients with active ulcerative colitis (UC). However, the cellular sources of miR-21 expression in IBD tissues have not yet been identified. We sought to determine the expression levels of miR-21 and one of its downstream target genes, programmed cell death 4 (PDCD4), in CD3 T cells isolated from the colonic mucosa of patients with active IBD, inactive IBD, and non-IBD controls...
November 7, 2016: Inflammatory Bowel Diseases
https://www.readbyqxmd.com/read/27818247/gapdh-binds-akt-to-facilitate-cargo-transport-in-the-early-secretory-pathway
#20
Ellen J Tisdale, Nikunj K Talati, Cristina R Artalejo, Assia Shisheva
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) undergoes numerous post-translational modifications, which impart new function and influence intracellular location. For example, atypical PKC ι/λ phosphorylates GAPDH that locates to vesicular tubular clusters and is required for retrograde membrane trafficking in the early secretory pathway. GAPDH is also required in the endocytic pathway; substitution of Pro(234) to Ser (Pro(234)Ser) rendered CHO cells defective in endocytosis. To determine if GAPDH (Pro(234)Ser) could inhibit endoplasmic reticulum to Golgi trafficking, we introduced the recombinant mutant enzyme into several biochemical and morphological transport assays...
December 10, 2016: Experimental Cell Research
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