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https://www.readbyqxmd.com/read/29236258/using-tet-off-cells-and-rnai-knockdown-to-assay-mrna-decay
#1
Thomas D Baird, J Robert Hogg
Cellular mRNA levels are determined by the competing forces of transcription and decay. A wide array of cellular mRNA decay pathways carry out RNA turnover either on a constitutive basis or in response to changing cellular conditions. Here, we outline a method to investigate mRNA decay that employs RNAi knockdown of known or putative decay factors in commercially available Tet-off cell systems. Reporter mRNAs of interest are expressed under the control of a tetracycline-regulated promoter, allowing pulse-chase mRNA decay assays to be conducted...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29225014/leber-s-hereditary-optic-neuropathy-caused-by-a-mutation-in-mitochondrial-trnathr-in-eight-chinese-pedigrees
#2
Juanjuan Zhang, Yanchun Ji, Xiaoling Liu, Jie Chen, Bibin Wang, Minglian Zhang, Min-Xin Guan
PURPOSE: The purpose of this study was to investigate the pathophysiology underlying Leber's hereditary optic neuropathy (LHON)-associated mitochondrial tRNA mutation. METHODS: Severn hundred ninety-seven Han Chinese subjects underwent clinical and genetic evaluation and analysis of mitochondrial DNA (mtDNA). The cybrid cell lines were constructed by transferring mitochondria from lymphoblastoid cell lines derived from a Chinese family into mtDNA-less (ρo) cells...
December 7, 2017: Mitochondrion
https://www.readbyqxmd.com/read/29216945/-expression-and-meaning-of-t-bet-and-gata3-mrna-in-t-lymphocyte-of-patients-during-the-operation-with-cardiopulmonary-bypass
#3
Haifeng Ni, Yingbin Xiao
OBJECTIVE: To investigate the effects of cardiopulmonary bypass (CPB) on the differentiation of T lymphocyte subsets and the expression of specific transcription regulator T-bet/GATA binding protein 3 (GATA3). METHODS: A prospective double-blind study was conducted. Patients with CPB pulmonary repair of ventricular septal defect (observation group) or off-pump ligation of ductus arteriosus (control group) with 20 cases each in the 150th Military Hospital from February 2015 to February 2016 were enrolled...
December 2017: Zhonghua Wei Zhong Bing Ji Jiu Yi Xue
https://www.readbyqxmd.com/read/29210354/neurocysticercosis-among-people-living-nearby-pigs-heavily-infected-with-cysticercosis-in-rural-endemic-peru
#4
Brian T Garvey, Luz M Moyano, Viterbo Ayvar, Silvia Rodriguez, Robert H Gilman, Armando E Gonzalez, Hector H Garcia, Seth E O'Neal, For The Cysticercosis Working Group In Peru
Neurocysticercosis causes substantial neurologic morbidity in endemic regions around the world. In this cross-sectional study, we describe the frequency of neurocysticercosis among a presumed high-risk group of people in an endemic community in northern Peru. Participants who screened positive on a nine-question seizure survey were evaluated clinically to diagnose epilepsy using International League Against Epilepsy criteria. Those with epilepsy were offered a noncontrast computerized tomography (CT) of the head...
December 4, 2017: American Journal of Tropical Medicine and Hygiene
https://www.readbyqxmd.com/read/29197505/a-safer-urea-based-in-situ-hybridization-method-improves-detection-of-gene-expression-in-diverse-animal-species
#5
Chiara Sinigaglia, Daniel Thiel, Andreas Hejnol, Evelyn Houliston, Lucas Leclère
In situ hybridization is a widely employed technique allowing spatial visualization of gene expression in fixed specimens. It has greatly advanced our understanding of biological processes, including developmental regulation. In situ protocols are today routinely followed in numerous laboratories, and although details might change, they all include a hybridization step, where specific antisense RNA or DNA probes anneal to the target nucleic acid sequence. This step is generally carried out at high temperatures and in a denaturing solution, called hybridization buffer, commonly containing 50% (v/v) formamide - a hazardous chemical...
December 8, 2017: Developmental Biology
https://www.readbyqxmd.com/read/29193387/the-nuclear-hormone-receptor-e75a-regulates-vitellogenin-gene-al-vg-expression-in-the-mirid-bug-apolygus-lucorum
#6
Y-A Tan, X-D Zhao, Y Sun, D-J Hao, J Zhao, Y-P Jiang, L-X Bai, L-B Xiao
Apolygus lucorum is the predominant pest of Bacillus thuringiensis (Bt) cotton in China. 20-hydroxyecdysone (20E) plays a key role in the reproduction of this insect. To better understand the mechanism underlying 20E-regulated reproduction, the nuclear hormone receptor E75 isoform-A of Ap. lucorum (Al-E75A) was cloned and its expression analysed. A 2241-bp sequence of Al-E75A cDNA encoded an open reading frame of a polypeptide with a predicted molecular mass of 69.04 kDa. Al-E75A mRNA was detected in female adult stages of Ap...
November 29, 2017: Insect Molecular Biology
https://www.readbyqxmd.com/read/29179278/-expression-of-macrophage-migration-inhibitory-factor-gene-in-placenta-tissue-and-its-correlation-with-gestational-diabetes-mellitus
#7
L Zheng, C Li, W H Qi, B L Qiao, H Zhao, Y B Zhou, C X Lu
Objective: To investigate the relationship between the expression of macrophage migration inhibitory factor (MIF) gene and the incidence of gestational diabetes mellitus (GDM) in Chinese Han population in northern China. Method: This study was consisted of 126 GDM women and 150 cases of healthy subjects who came from the obstetrics department of Maternity Hospital of Qingdao University and Maternity and Child Care Hospital of Donggang District of Rizhao. The protein expression of MIF in placenta tissue was detected by Western blot method...
November 21, 2017: Zhonghua Yi Xue za Zhi [Chinese medical journal]
https://www.readbyqxmd.com/read/29176795/the-full-transcription-map-of-mouse-papillomavirus-type-1-mmupv1-in-mouse-wart-tissues
#8
Xiang-Yang Xue, Vladimir Majerciak, Aayushi Uberoi, Bong-Hyun Kim, Deanna Gotte, Xiongfong Chen, Maggie Cam, Paul F Lambert, Zhi-Ming Zheng
Mouse papillomavirus type 1 (MmuPV1) provides, for the first time, the opportunity to study infection and pathogenesis of papillomaviruses in the context of laboratory mice. In this report, we define the transcriptome of MmuPV1 genome present in papillomas arising in experimentally infected mice using a combination of RNA-seq, PacBio Iso-seq, 5' RACE, 3' RACE, primer-walking RT-PCR, RNase protection, Northern blot and in situ hybridization analyses. We demonstrate that the MmuPV1 genome is transcribed unidirectionally from five major promoters (P) or transcription start sites (TSS) and polyadenylates its transcripts at two major polyadenylation (pA) sites...
November 27, 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/29175328/construction-of-a-high-yield-dengue-virus-by-replacing-nonstructural-proteins-3-4b-without-increasing-virulence
#9
Supranee Phanthanawiboon, Sabar Pambudi, Magot Diata Omokoko, Keiko Hanabara, Atchareeya A-Nuegoonpipat, Wataru Kamitani, Kazuyoshi Ikuta, Takeshi Kurosu
Producing virus at high yield is critically important for development of whole virion inactivated vaccines or live attenuated vaccines. Most dengue virus (DENV) clinical isolates, however, replicate at low levels in cultured cells, which limits their use for vaccine development. The present study examined differences between low-replicating DENV clinical isolates and high-replicating laboratory strains with the aim of engineering high-yield DENV clinical isolates. Construction of a series of recombinant chimeric viruses derived from a high-replicating laboratory DENV type 4 (DENV-4) H241 strain and a clinical isolate revealed that the NS3-NS4B region of H241 conferred a replication advantage in cultured cells...
November 21, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29163430/pdtas-deficiency-affects-resistance-of-mycobacteria-to-ribosome-targeting-antibiotics
#10
Karolina Dadura, Renata Płocińska, Anna Rumijowska-Galewicz, Przemysław Płociński, Anna Żaczek, Bożena Dziadek, Andrzej Zaborowski, Jarosław Dziadek
Two-component regulatory systems (TCSSs) are key regulatory elements responsible for the adaptation of bacteria to environmental stresses. A classical TCSS is typically comprised of a sensory histidine kinase and a corresponding response regulator. Here, we used homologous recombination to construct a Mycobacterium smegmatis mutant defective in the synthesis of cytosolic histidine kinase PdtaS (Msmeg_1918). The resulting ΔpdtaS mutant strain was tested in the Phenotype Microarray screening system, which allowed us to identify aminoglycoside antibiotic sensitivity, tetracyclines antibiotic resistance as well as membrane transport and respiration, as the main processes affected by removal of pdtaS...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/29153168/a-100-year-review-mammary-development-and-lactation
#11
R Michael Akers
What is old is new again-and with respect to the study of the mammary development and function in dairy animals, the expression resonates. Many of the mammary and milk production questions raised in the early years of the Journal of Dairy Science apply today. To be sure, scientists have filled in many details regarding, for example, identification of hormones and growth factors important in the control of mammary growth, the onset of copious milk production at calving, and maintenance of lactation. Early years focused on identification and subsequent availability of classic mammogenic, lactogenic, and galactopoietic hormones (e...
December 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/29129493/overexpression-of-microrna-495-improves-the-intestinal-mucosal-barrier-function-by-targeting-stat3-via-inhibition-of-the-jak-stat3-signaling-pathway-in-a-mouse-model-of-ulcerative-colitis
#12
Xian-Qun Chu, Jing Wang, Guang-Xiang Chen, Guan-Qi Zhang, De-Yong Zhang, Yong-Yan Cai
We aim to investigate the role of microRNA-495 (miR-495) in the intestinal mucosal barrier by indirectly targeting signal transducer and activator of transcription 3 (STAT3) through the Janus kinase-signal transducer and activator of transcription (JAK)/STAT3 signaling pathway in a mouse model of ulcerative colitis (UC). BALB/c mice were selected for establishing mice model of UC, and intestinal tissues of normal and UC mice were collected. ELISA was conducted for detecting levels of TNF-α, IL-6, IFN-γ and IL-10...
October 12, 2017: Pathology, Research and Practice
https://www.readbyqxmd.com/read/29109187/homologous-vapc-toxins-inhibit-translation-and-cell-growth-by-sequence-specific-cleavage-of-trna-fmet
#13
Lauren R Walling, J Scott Butler
Type II toxin-antitoxin (TA) systems play a critical role in the establishment and maintenance of bacterial dormancy. They are composed of a protein toxin and its cognate protein antitoxin. They function to regulate growth during conditions of stress, such as starvation or antibiotic treatment. As cellular proteases degrade the antitoxin, which normally binds and neutralizes the toxin, this frees the toxin to act on its cellular targets and arrest bacterial growth. TA systems are of particular concern in pathogenic organisms such as nontypeable Haemophilus influenzae (NTHi), as dormancy may lead to chronic infections and failure of antibiotic treatment...
November 6, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/29108629/spontaneous-pepsin-c-catalyzed-activation-of-human-pepsinogen-c-in-transgenic-rice-cell-suspension-culture-production-and-characterization-of-human-pepsin-c
#14
Md Reyazul Islam, Nan-Sun Kim, Jae-Wan Jung, Hyo-Boon Kim, So-Chon Han, Moon-Sik Yang
A human pepsinogen C (hPGC) gene was synthesized with rice-optimized codon usage and cloned into a rice expression vector containing the promoter, signal peptide, and terminator derived from the rice α-amylase 3D (Ramy3D) gene. In addition, a 6-His tag was added to the 3' end of the synthetic hPGC gene for easy purification. The plant expression vector was introduced into rice calli (Oryza sativa L. cv. Dongjin) mediated by Agrobacterium tumefaciens. The integration of the hPGC gene into the chromosome of the transgenic rice callus and hPGC expression in transgenic rice cell suspensions was verified via genomic DNA polymerase chain reaction amplification and Northern blot analysis...
January 2018: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/29099311/a-comparative-study-of-sequence-and-structure-based-features-of-small-rnas-and-other-rnas-of-bacteria
#15
Amita Barik, Santasabuj Das
Small RNAs (sRNAs) in bacteria have emerged as key players in transcriptional and post-transcriptional regulation of gene expression. Here, we present a statistical analysis of different sequence- and structure-related features of bacterial sRNAs to identify the descriptors that could discriminate sRNAs from other bacterial RNAs. We investigated a comprehensive and heterogeneous collection of 816 sRNAs, identified by northern blotting across 33 bacterial species and compared their various features with other classes of bacterial RNAs, such as tRNAs, rRNAs and mRNAs...
November 3, 2017: RNA Biology
https://www.readbyqxmd.com/read/29084267/overexpression-of-arabidopsis-thaliana-brassinosteroid-related-acyltransferase-1-gene-induces-brassinosteroid-deficient-phenotypes-in-creeping-bentgrass
#16
Yun-Jeong Han, Young Soon Kim, Ok-Jin Hwang, Jeehee Roh, Keya Ganguly, Seong-Ki Kim, Ildoo Hwang, Jeong-Il Kim
Brassinosteroids (BRs) are naturally occurring steroidal hormones that play diverse roles in various processes during plant growth and development. Thus, genetic manipulation of endogenous BR levels might offer a way of improving the agronomic traits of crops, including plant architecture and stress tolerance. In this study, we produced transgenic creeping bentgrass (Agrostis stolonifera L.) overexpressing a BR-inactivating enzyme, Arabidopsis thaliana BR-related acyltransferase 1 (AtBAT1), which is known to catalyze the conversion of BR intermediates to inactive acylated conjugates...
2017: PloS One
https://www.readbyqxmd.com/read/29082339/dnajc18-a-novel-type-iii-dnaj-family-protein-is-expressed-specifically-in-rat-male-germ-cells
#17
Cynthia Gomes, Jaemog Soh
Mammalian spermatogenesis occurs in a precise and coordinated manner in the seminiferous tubules. One of the attempts to understand the detailed biological process during mammalian spermatogenesis at the molecular level has been to identify the testis specific genes followed by study of the testicular expression pattern of the genes. From the subtracted cDNA library of rat testis prepared using representational difference analysis (RDA) method, a complimentary DNA clone encoding type III member of a DnaJ family protein, DnaJC18, was cloned (GenBank Accession No...
September 2017: Balsaeng'gwa Saengsig
https://www.readbyqxmd.com/read/29077738/rnai-derived-transgenic-resistance-to-mungbean-yellow-mosaic-india-virus-in-cowpea
#18
Sanjeev Kumar, Bhaben Tanti, Basavaprabhu L Patil, Sunil Kumar Mukherjee, Lingaraj Sahoo
Cowpea is an important grain legume crop of Africa, Latin America, and Southeast Asia. Leaf curl and golden mosaic diseases caused by Mungbean yellow mosaic India virus (MYMIV) have emerged as most devastating viral diseases of cowpea in Southeast Asia. In this study, we employed RNA interference (RNAi) strategy to control cowpea-infecting MYMIV. For this, we generated transgenic cowpea plants harbouring three different intron hairpin RNAi constructs, containing the AC2, AC4 and fusion of AC2 and AC4 (AC2+AC4) of seven cowpea-infecting begomoviruses...
2017: PloS One
https://www.readbyqxmd.com/read/29071477/enforced-expression-of-mir142-a-target-of-chromosome-translocation-in-human-b-cell-tumors-results-in-b-cell-depletion
#19
Kodai Kuriyama, Yutaka Enomoto, Ritsuro Suzuki, Jyuri Watanuki, Hiroki Hosoi, Yusuke Yamashita, Shogo Murata, Toshiki Mushino, Shinobu Tamura, Nobuyoshi Hanaoka, Martin Dyer, Reiner Siebert, Hiroshi Kiyonari, Hideki Nakakuma, Toshio Kitamura, Takashi Sonoki
MicroRNA142 (MIR142) is a target of chromosome translocations and mutations in human B-cell lymphomas. We analyzed an aggressive B-cell lymphoma carrying t(8;17)(q24;q22) and t(6;14)(p21;q32), and sought to explore the role(s) of MIR142 in lymphomagenesis. t(8;17)(q24;q22) involved MYC on 8q24 and pri-MIR142 on 17q22. MYC was activated by a promoter substitution by t(8;17)(q24;q22). t(8;17)(q24;q22) was an additional event after t(6;14) (p21;q32), which caused the over-expression of CCND3. Southern blot analyses revealed that the MIR142 locus was deleted from the affected allele, whereas Northern analyses showed over-expression of MIR142 in tumor cells...
October 25, 2017: International Journal of Hematology
https://www.readbyqxmd.com/read/29062054/integrated-analysis-of-mrna-and-viral-mirnas-in-the-kidney-of-carassius-auratus-gibelio-response-to-cyprinid-herpesvirus-2
#20
Jianfei Lu, Dan Xu, Yousheng Jiang, Shanyun Kong, Zhaoyuan Shen, Siyao Xia, Liqun Lu
MicroRNAs (miRNAs) are small, non-coding single stranded RNAs that play crucial roles in numerous biological processes. Vertebrate herpesviruses encode multiple viral miRNAs that modulate host and viral genes. However, the roles of viral miRNAs in lower vertebrates have not been fully determined. Here, we used high-throughput sequencing to analyse the miRNA and mRNA expression profiles of Carassius auratus gibelio in response to infection by cyprinid herpesvirus 2 (CyHV-2). RNA sequencing obtained 26,664 assembled transcripts, including 2,912 differentially expressed genes...
October 23, 2017: Scientific Reports
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