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Quantitative RT-PCR

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https://www.readbyqxmd.com/read/28938749/the-testicular-transcriptome-associated-with-spermatogonia-differentiation-initiated-by-gonadotrophin-stimulation-in-the-juvenile-rhesus-monkey-macaca-mulatta
#1
Suresh Ramaswamy, William H Walker, Paula Aliberti, Rahil Sethi, Gary R Marshall, Alyxzandria Smith, Seyedmehdi Nourashrafeddin, Alicia Belgorosky, Uma R Chandran, Mark P Hedger, Tony M Plant
STUDY QUESTION: What is the genetic landscape within the testis of the juvenile rhesus monkey (Macaca mulatta) that underlies the decision of undifferentiated spermatogonia to commit to a pathway of differentiation when puberty is induced prematurely by exogenous LH and FSH stimulation? SUMMARY ANSWER: Forty-eight hours of gonadotrophin stimulation of the juvenile monkey testis resulted in the appearance of differentiating B spermatogonia and the emergence of 1362 up-regulated and 225 down-regulated testicular mRNAs encoding a complex network of proteins ranging from enzymes regulating Leydig cell steroidogenesis to membrane receptors, and from juxtacrine and paracrine factors to transcriptional factors governing spermatogonial stem cell fate...
October 1, 2017: Human Reproduction
https://www.readbyqxmd.com/read/28938650/genomic-variants-link-to-hepatitis-c-racial-disparities
#2
Matthew M Yeh, Sarag Boukhar, Benjamin Roberts, Nairanjana Dasgupta, Sayed S Daoud
Chronic liver diseases are one of the major public health issues in United States, and there are substantial racial disparities in liver cancer-related mortality. We previously identified racially distinct alterations in the expression of transcripts and proteins of hepatitis C (HCV)-induced hepatocellular carcinoma (HCC) between Caucasian (CA) and African American (AA) subgroups. Here, we performed a comparative genome-wide analysis of normal vs. HCV+ (cirrhotic state), and normal adjacent tissues (HCCN) vs...
August 29, 2017: Oncotarget
https://www.readbyqxmd.com/read/28938580/diagnostic-significance-of-urinary-long-non-coding-pca3-rna-in-prostate-cancer
#3
Tao Wang, Xiangyun Qu, Jiajia Jiang, Peng Gao, Dingding Zhao, Xueqi Lian, Xiaohua Li
Prostate cancer antigen 3(PCA3) is a long non-coding RNA, which was found increased expression in CaP patients than healthy individual. In this study, the individual nucleic acid of PCA3 and PSA was recombinant expressed as a reference reagent, and a quantitative RT-PCR with TaqMan assay was developed to examine the copies of PCA3 and PSA gene in urine. The results showed that the area under the receiver operating characteristic curve (AUROC) was 0.717, 0.444 and 0.916 for the number of PCA3 copy, PSA copy and for the score of PCA3/PSA RNA, respectively...
August 29, 2017: Oncotarget
https://www.readbyqxmd.com/read/28938535/accurate-primary-germ-cell-cancer-diagnosis-using-serum-based-microrna-detection-amptsmir-test
#4
Ton van Agthoven, Leendert H J Looijenga
Multiple studies, including various methods and overall limited numbers of mostly heterogeneous cases, indicate that the level of embryonic stem cell microRNAs (miRs) (e.g. 371a-3p, 372-3p, 373-3p, and 367-3p) are increased in serum at primary diagnosis of almost all testicular germ cell cancer (TGCC). Here we determine the status of three of these miRs in serum samples of 250 TGCC patients, collected at time of primary diagnosis, compared with 60 non-TGCC patients and 104 male healthy donors. The levels of miRs were measured by the robust ampTSmiR test, including magnetic bead-based miR isolation and target specific pre-amplification followed by real-time quantitative PCR (RT-qPCR) detection...
August 29, 2017: Oncotarget
https://www.readbyqxmd.com/read/28938428/glp-1-receptor-activation-inhibits-palmitate-induced-apoptosis-via-ceramide-in-human-cardiac-progenitor-cells
#5
Anna Leonardini, Rossella D'Oria, Maria Angela Incalza, Cristina Caccioppoli, Valentina Andrulli Buccheri, Angelo Cignarelli, Domenico Paparella, Vito Margari, Annalisa Natalicchio, Sebastio Perrini, Francesco Giorgino, Luigi Laviola
Context: Increased apoptosis of cardiomyocytes and cardiac progenitor cells (CPCs) in response to saturated fatty acids (SFA) can lead to myocardial damage and dysfunction. Ceramides mediate lipotoxicity-induced apoptosis. GLP-1 receptor (GLP1R) agonists exert beneficial effects on cardiac cells in experimental models. Objective: To investigate the protective effects of GLP1R activation on SFA-mediated apoptotic death of human CPCs. Design: Human CPCs were isolated from cardiac appendages of non-diabetic donors and then exposed to palmitate with or without pre-treatment with the GLP1R agonist exendin-4...
August 16, 2017: Journal of Clinical Endocrinology and Metabolism
https://www.readbyqxmd.com/read/28935572/macrophage-infiltration-is-a-causative-factor-for-ligamentum-flavum-hypertrophy-through-the-activation-of-collagen-production-in-fibroblasts
#6
Takeyuki Saito, Masamitsu Hara, Hiromi Kumamaru, Kazu Kobayakawa, Kazuya Yokota, Ken Kijima, Shingo Yoshizaki, Katsumi Harimaya, Yoshihiro Matsumoto, Kenichi Kawaguchi, Mitsumasa Hayashida, Yutaka Inagaki, Keiichiro Shiba, Yasuharu Nakashima, Seiji Okada
Ligamentum flavum (LF) hypertrophy causes lumbar spinal canal stenosis, leading to leg pain and disability in activities of daily living in elderly individuals. Although there have been previous studies on LF hypertrophy, its pathomechanisms have not been fully elucidated. In this study, we demonstrated that infiltrating macrophages were a causative factor for LF hypertrophy. Induction of macrophages into the mouse LF by applying a micro-injury resulted in LF hypertrophy along with collagen accumulation and fibroblasts proliferation at the injured site, which were very similar to the characteristics observed in the severely hypertrophied LF of human...
September 18, 2017: American Journal of Pathology
https://www.readbyqxmd.com/read/28932224/shrimp-mir-10a-is-co-opted-by-white-spot-syndrome-virus-to-increase-viral-gene-expression-and-viral-replication
#7
Jiun-Yan Huang, Shih-Ting Kang, I-Tung Chen, Li-Kwan Chang, Shih-Shun Lin, Guang-Hsiung Kou, Chia-Ying Chu, Chu-Fang Lo
Members of the microRNA miR-10 family are highly conserved and play many important roles in diverse biological mechanisms, including immune-related responses and cancer-related processes in certain types of cancer. In this study, we found the most highly upregulated shrimp microRNA from Penaeus vannamei during white spot syndrome virus (WSSV) infection was miR-10a. After confirming the expression level of miR-10a by northern blot and quantitative RT-PCR, an in vivo experiment showed that the viral copy number was decreased in miR-10a-inhibited shrimp...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28931690/conformational-changes-in-the-5-end-of-the-hiv-1-genome-dependent-on-the-debranching-enzyme-dbr1-during-early-stages-of-infection
#8
Alvaro E Galvis, Hugh E Fisher, Hung Fan, David Camerini
Previous studies in our laboratory showed that the RNA debranching enzyme (DBR1) is not required for early steps in HIV cDNA formation, but is necessary for synthesis of intermediate and late cDNA products. To further characterize this effect, we evaluated the topology of the 5' end of the HIV-1 RNA genome during early infection with and without inhibition of DBR1 synthesis. Cells were transfected with DBR1 shRNA followed 48 hours later by infection with an HIV-1 derived vector containing an RNase H deficient reverse transcriptase...
September 20, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28931396/il-4-induced-caveolin-1-containing-lipid-rafts-aggregation-contributes-to-muc5ac-synthesis-in-bronchial-epithelial-cells
#9
Yu Xia, Peng-Cheng Cai, Fan Yu, Liang Xiong, Xin-Liang He, Shan-Shan Rao, Feng Chen, Xiang-Ping Yang, Wan-Li Ma, Hong Ye
BACKGROUND: Mucus overproduction is an important feature of asthma. Interleukin (IL)-4 is required for allergen-induced airway inflammation and mucus production. MUC5AC gene expression is regulated by transcript factors NF-κB. The intracellular Ca(2+) ([Ca(2+)]i) signal is required for activation of NF-κB. The transient receptor potential canonical 1 (TRPC1) channel has been shown to contribute for agonist-stimulated Ca(2+) influx in some types of cells. However, the relationships among IL-4, TRPC1 and mucus overproduction in bronchial epithelial cells (BECs) in asthma are poorly understood...
September 20, 2017: Respiratory Research
https://www.readbyqxmd.com/read/28929648/-cloning-and-expression-analysis-of-tocopherol-cyclase-gene-in-carthamus-tinctorius
#10
Li-Li Guan, Qi Cui, Yu-Tong Han, Yun-Yun Wu, Ren-Ge Hu, Tian-Yao Gu, Hai-Yan Li, Xiao-Kun Li
The tocopherol cyclase was one of the key enzymes in plant vitamin E biosynthesis pathway. According to the study of Carthamus tinctorius transcriptome data,the Tocopherol cyclase gene was obtained using RT-PCR techniques and named CtTC . Bioinformatics analysis showed theopen reading frame (ORF)of CtTC was 1 524 bp. The putative protein contained 507 amino acids with a predicted molecular mass of 62.9 kDa and theoretically isoelectric point was 5.01.Signal peptide analysis showed that it was a non secretory protein, and there was no signal peptide...
October 2016: Zhongguo Zhong Yao za Zhi, Zhongguo Zhongyao Zazhi, China Journal of Chinese Materia Medica
https://www.readbyqxmd.com/read/28929270/mode-of-treatment-governs-curcumin-response-on-doxorubicin-induced-toxicity-in-cardiomyoblasts
#11
Aditi Jain, Vibha Rani
Doxorubicin (Dox) is an effective anti-cancer drug with severe reported cardiotoxicity. Cardiovascular risks associated with present cancer therapeutics demand urgent attention. There has been a growing interest in naturally occurring compounds to improve the therapeutic index as well as prevent non-tumour tissues from sustaining chemotherapy-induced damages. In the present study, the effects of curcumin, a polyphenol isolated from Curcuma longa and well known for its anti-oxidative, anti-cancerous and anti-inflammatory properties, was studied in relation to the Dox-induced cardiotoxicity...
September 19, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28929229/a-novel-method-for-concentration-of-porcine-reproductive-and-respiratory-syndrome-virus-from-the-environmental-samples-using-self-aggregating-peptide-tagged-cd151-binding-capture
#12
Yang Zong, Xiaoyin Zong, Wenlong Xia, Zhi Wu, Guangya Li, Yangyang Li, Xinyu Zhang, Xiaoli Xia, Huaichang Sun
Environmental surveillance of porcine reproductive and respiratory syndrome virus (PRRSV) represents a key issue in control of the disease. CD151 has recently been recognized as one of several receptors for PRRSV. We describe here a novel method for concentration of PRRSV from the environmental samples by CD151-binding capture. After fusion to self-aggregating peptide ELK16, the large extracellular loop (LEL) of porcine CD151 and its two segments (namely N63 and C63) were expressed in E. coli as protein aggregates...
September 19, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28928821/involvement-of-estrogen-receptor-%C3%AE-in-androgen-receptor-induced-growth-inhibition-in-prostate-cancer-pc-3-cells
#13
Long Xiao, Minhui Xiao, Linbo Gao, Wanchao Xu
Previous studies have suggested that changes in sex hormone receptor expression may be associated with the initiation and progression of prostate cancer (PCa). Therefore, the present study aimed to investigate the association and possible pathways between two sex hormone receptors and PCa by measuring the expression levels of the androgen receptor (AR) and the estrogen receptor subtypes alpha (ERα) and beta (ERβ) in prostatic cancer PC-3 cell lines. The pcDNA3.1-hERβ plasmid was transfected into PC-3 cell lines...
September 2017: Oncology Letters
https://www.readbyqxmd.com/read/28926893/-effect-of-intratumor-heterogeneity-of-esophageal-squamous-cell-carcinoma-on-chemotherapy-sensitivity
#14
L Sun, W Wu, M Yan, P L Han, X Zhan, X W Ma, X G Cao, S Zhao, F Gao, Y Qi, W Cao
Objective: To investigate the relationship of heterogeneity of esophageal squamous cell carcinoma (ESCC) and chemotherapy sensitivity. Methods: Five different region specimens isolated from primary tumor(R1~R5)and 1 specimen(R6)isolated from adjacent non-neoplastic tissue from 10 ESCC patients who underwent surgical treatment were cultured in vitro. The inhibitory effect of cisplatin on proliferation of ESCC cells from different regions was determined by methyl thiazolyl tetrazolium (MTT). The cell cycle and apoptosis induced by cisplatin was determined by flow cytometry (FCM) analysis...
September 23, 2017: Zhonghua Zhong Liu za Zhi [Chinese Journal of Oncology]
https://www.readbyqxmd.com/read/28926609/developmental-expression-of-membrane-type-4-matrix-metalloproteinase-mt4-mmp-mmp17-in-the-mouse-embryo
#15
María José Blanco, Iván Rodríguez-Martín, Ana I R Learte, Cristina Clemente, María Gregoria Montalvo, Motoharu Seiki, Alicia G Arroyo, Cristina Sánchez-Camacho
Matrix metalloproteinases (MMPs) constitute a large group of endoproteases that play important functions during embryonic development, tumor metastasis and angiogenesis by degrading components of the extracellular matrix. Within this family, we focused our study on Mt4-mmp (also called Mmp17) that belongs to a distinct subset that is anchored to the cell surface via a glycosylphosphatidylinositol (GPI) moiety and with the catalytic site exposed to the extracellular space. Information about its function and substrates is very limited to date, and little has been reported on its role in the developing embryo...
2017: PloS One
https://www.readbyqxmd.com/read/28926402/reversing-hiv-latency-via-sphingosine-1-phosphate-receptor-1-signaling
#16
Charline Duquenne, Sandrine Gimenez, Adeline Guigues, Benjamin Viala, Caroline Boulouis, Clément Mettling, Damien Maurel, Noëlie Campos, Etienne Doumazane, Laetitia Comps-Agrar, Jamal Tazi, Laurent Prézeau, Pierre Corbeau, Vincent François
OBJECTIVE: In this study, we looked for a new family of latency reversing agents. DESIGN: We searched for G protein-coupled receptors (GPCR) coexpressed with CCR5 in primary CD4+ T cells that activate infected cells and boost HIV production. METHODS: GPCR coexpression was unveiled by RT-PCR. We used FRET to analyze the dimerization with CCR5 of the expressed GPCR. Viral entry was measured by flow cytometry, reverse transcription by quantitative PCR, NFkB translocation by immunofluorescence, LTR activation using a gene reporter assay, and viral production by p24 quantification...
September 18, 2017: AIDS
https://www.readbyqxmd.com/read/28923591/transcriptome-profiling-based-on-protein-protein-interaction-networks-provides-a-core-set-of-genes-for-understanding-blood-immune-response-mechanisms-against-edwardsiella-tarda-infection-in-japanese-flounder-paralichthys-olivaceus
#17
Zan Li, Xiumei Liu, Jinxiang Liu, Kai Zhang, Haiyang Yu, Yan He, Xubo Wang, Jie Qi, Zhigang Wang, Quanqi Zhang
Marine organisms are commonly under threat from various pathogens. Edwardsiella tarda is one of the fish pathogens that can infect both cultured and wild fish species. E. tarda can also infect other vertebrates, including amphibians, reptiles, and mammals. Bacteremia caused by E. tarda can be a fatal disease in humans. Blood acts as a pipeline for the fish immune system. Generating blood transcriptomic resources from fish challenged by E. tarda is crucial for understanding molecular mechanisms underlying blood immune response process...
September 15, 2017: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/28923068/using-blood-cytokine-measures-to-define-high-inflammatory-biotype-of-schizophrenia-and-schizoaffective-disorder
#18
Danny Boerrigter, Thomas W Weickert, Rhoshel Lenroot, Maryanne O'Donnell, Cherrie Galletly, Dennis Liu, Martin Burgess, Roxanne Cadiz, Isabella Jacomb, Vibeke S Catts, Stu G Fillman, Cynthia Shannon Weickert
BACKGROUND: Increases in pro-inflammatory cytokines are found in the brain and blood of people with schizophrenia. However, increased cytokines are not evident in all people with schizophrenia, but are found in a subset. The cytokine changes that best define this subset, termed the "elevated inflammatory biotype", are still being identified. METHODS: Using quantitative RT-PCR, we measured five cytokine mRNAs (IL-1β, IL-2 IL-6, IL-8 and IL-18) from peripheral blood of healthy controls and of people with schizophrenia or schizoaffective disorder (n = 165)...
September 18, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28922848/aggressive-behavior-in-silent-subtype-iii-pituitary-adenomas-may-depend-on-suppression-of-local-immune-response-a-whole-transcriptome-analysis
#19
Timothy E Richardson, Zhong-Jian Shen, Mohammed Kanchwala, Chao Xing, Alexander Filatenkov, Ping Shang, Samuel Barnett, Zahidur Abedin, James S Malter, Jack M Raisanen, Dennis K Burns, Charles L White, Kimmo J Hatanpaa
Silent subtype III pituitary adenomas (SS-3) are clinically nonfunctional adenomas that are more aggressive in terms of invasion and risk of recurrence than their conventional null cell counterparts. We previously showed that these tumors can be distinguished by immunohistochemistry based on the identification of a markedly enlarged and fragmented Golgi apparatus. To understand the molecular correlates of differential aggressiveness, we performed whole transcriptome sequencing (RNAseq) on 4 SS-3 and 4 conventional null cell adenomas...
October 1, 2017: Journal of Neuropathology and Experimental Neurology
https://www.readbyqxmd.com/read/28922680/pivotal-role-of-tnf-%C3%AE-in-the-development-and-progression-of-nonalcoholic-fatty-liver-disease-in-a-murine-model
#20
Satomi Kakino, Tsuyoshi Ohki, Hitomi Nakayama, Xiahong Yuan, Shuichi Otabe, Toshihiko Hashinaga, Nobuhiko Wada, Yayoi Kurita, Kayo Tanaka, Kento Hara, Eri Soejima, Yuji Tajiri, Kentaro Yamada
Previously, we have shown that the adipocyte-specific nuclear form of sterol regulatory element-binding protein-1c (nSREBP-1c) transgenic mice spontaneously developed hepatic lesions that are similar to those of human nonalcoholic steatohepatitis (NASH) with a concomitant elevation of plasma TNF-α. In this study, we analyzed the role of TNF-α in the progression of nonalcoholic fatty liver disease (NAFLD). We established a Tnf knockout nSREBP-1c transgenic mouse line. Glucose tolerance and liver histology were examined at the age of 20 weeks...
September 18, 2017: Hormone and Metabolic Research, Hormon- und Stoffwechselforschung, Hormones et Métabolisme
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