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GeXP genetic analysis system

Ivan Kutyrev, Beth Cleveland, Timothy Leeds, Gregory D Wiens
Flavobacterium psychrophilum (Fp) is the causative agent of bacterial cold water disease (BCWD) which causes appreciable economic losses in rainbow trout aquaculture. We previously reported development of a genetic line, designated ARS-Fp-R that exhibits higher survival relative to a susceptible line, designated ARS-Fp-S, following either laboratory or natural on-farm challenge. The objectives of this study were to determine the temporal kinetics of gene expression between experimentally-challenged ARS-Fp-R and ARS-Fp-S fish and the correlation between gene expression and pathogen load...
October 11, 2016: Fish & Shellfish Immunology
Takashi Morimoto, Gouranga Biswas, Tomoya Kono, Masahiro Sakai, Jun-Ichi Hikima
Studies on immune response to crystal silica in mammals indicate immune stimulation effect of environmental parameters including silica or asbestos, but there is no information on this aspect in lower vertebrates. Therefore, we examined expression of cytokine genes related to innate immunity in the Japanese pufferfish, Fugu (Takifugu rubripes) head kidney (HK) cells stimulated with particulate silica at 10 and 50 μg mL(-1). Expression of eleven cytokine genes was analyzed by the multiplex RT-PCR method (GenomeLab Genetic Analysis System, GeXPS; Beckman Coulter Inc...
February 2016: Fish & Shellfish Immunology
Soner Bilen, Gouranga Biswas, Shohei Otsuyama, Tomoya Kono, Masahiro Sakai, Jun-ichi Hikima
A cytosolic receptor complex called inflammasome is responsible for mounting inflammatory response by releasing pro-inflammatory cytokines, interleukin (IL)-1β and IL-18. However, inflammatory cascades mediated by the inflammasome are unknown in a lower vertebrate like fish. Therefore, in an in vitro experiment, in order to obtain a preliminary information, we conducted transcriptomic analysis of 18 cytokines including pro-inflammatory cytokines in the Japanese pufferfish (Takifugu rubripes) head kidney (HK) cells stimulated with an inflammasome-inducing agent, nigericin, and a combination of nigericin and LPS by a multiplex RT-PCR assay (GenomeLab Genetic Analysis System, GeXPS; Beckman Coulter Inc...
October 2014: Developmental and Comparative Immunology
Jing Tong, Shilpa Buch, Honghong Yao, Chengxiang Wu, Hsin-I Tong, Youwei Wang, Yuanan Lu
HIV-1 associated dementia remains a significant public health burden. Clinical and experimental research has shown that reduced levels of brain-derived neurotrophic factor (BDNF) may be a risk factor for neurological complications associated with HIV-1 infection. We are actively testing genetically modified macrophages for their possible use as the cell-based gene delivery vehicle for the central nervous system (CNS). It can be an advantage to use the natural homing/migratory properties of monocyte-derived macrophages to deliver potentially neuroprotective BDNF into the CNS, as a non-invasive manner...
2014: PloS One
Joan Blin, Zalinah Ahmad, Lekhraj Rampal S O Gyanchand Rampal, Norhafizah Mohtarrudin, Abdul Karim Haji Tajudin, Rohayu Shahar Adnan
Identifying susceptible genes associated with the pathogenesis of atherosclerosis (ATH) may contribute toward better management of this condition. This preliminary study was aimed at assessing the expression levels of 11 candidate genes, namely tumor protein (TP53), transforming growth factor, beta receptor II (TGFBR2), cysthathionenine-beta-synthase (CBS), insulin receptor substrate 1 (IRS1), lipoprotein lipase (LPL), methylenetetrahydrofolate reductase (MTHFR), thrombomodulin (THBD), lecithin-cholesterol acyltransferase (LCAT), matrix metallopeptidase 9 (MMP9), low density lipoprotein receptor (LDLR), and arachidonate 5-lipoxygenase-activating protein (ALOX5AP) genes associated with ATH...
2013: Genes & Genetic Systems
Maria A Nagel, Alexander Choe, Don Gilden, Vicki Traina-Dorge, Randall J Cohrs, Ravi Mahalingam
Simian varicella zoster virus (SVV) infection of non-human primates serves as a model to study varicella zoster virus (VZV) infection and pathogenesis in humans. While macroarray analysis detected all 69 predicted unique open reading frames (ORFs) in SVV-infected cells in culture, it lacked the sensitivity to detect the low-abundance transcripts expressed in latently infected monkey ganglia. Recently, a multiplex RT-PCR assay using the GenomeLab Genetic Analysis System (GeXPS) identified 10 VZV transcripts in latently-infected human ganglia...
October 2013: Journal of Virological Methods
G Biswas, H Korenaga, R Nagamine, H Takayama, S Kawahara, S Takeda, Y Kikuchi, B Dashnyam, T Kono, M Sakai
Cytokine responses in the Japanese pufferfish (Takifugu rubripes) head kidney (HK) cells to heat-killed lactic acid bacteria probiotics isolated from the Mongolian dairy products were investigated by transcriptomic examination. The HK cells were incubated with two heat-killed bacteria, namely Lactobacillus paracasei spp. paracasei (strain 06TCa22) and L. plantarum (strain 06CC2) and the responses of 16 cytokine genes at 0 (control), 1, 4, 8, 12, 24 and 48 h post-stimulation were assayed by multiplex RT-PCR analysis (GenomeLab Genetic Analysis System, GeXPS; Beckman Coulter, Inc...
May 2013: Fish & Shellfish Immunology
Bin He, Huan-Yu Wang, Chen Zhang, Miao Wang, Meng Qin, Ke-Xia Wang, Xue-Jun Ma
Multiplex reverse transcription-polymerase chain reaction (mRT-PCR) is currently available in virus detection and defined as the simultaneous amplification of two or more DNA/RNA targets in a single reaction vessel. In this study, we attempted to modify the conventional mRT-PCR technique on a basis of GenomeLab Genetic Analysis System (GeXP). Initially, we optimized the analytical validation of the GeXP analyzer and its design of workflow and simultaneously detected eight arboviruses that related to epidemic encephalitis by verifying the specificity of mRT-PCR with Japanese encephalitis virus(JEV) cell cultures and positive strains identified previously and determining the sensitivity with in vitro-transcribed RNA of serial dilutions...
January 2012: Bing du Xue Bao, Chinese Journal of Virology
Lin Xie, Naotsugu Ichimaru, Miwa Morita, Jiajie Chen, Ping Zhu, Jihong Wang, Peter Urbanellis, Itay Shalev, Shizuko Nagao, Atsushi Sugioka, Liang Zhong, Norio Nonomura, Shiro Takahara, Gary A Levy, Xiao-Kang Li
Here we examined whether the expression of a novel immunoregulatory gene set could be used to predict outcomes in murine models of rapamycin-induced cardiac tolerance, spontaneous hepatic tolerance, and cardiac rejection. The expression of the immunoregulatory gene set was assessed with the GeXP multiplex reverse-transcription polymerase chain reaction (RT-PCR) analysis system, and it was correlated to the pathological and biochemical parameters of the allografts. In rejecting cardiac grafts, the increased expression of an inflammatory set of genes, which included CD45, CD4, CD25, suppressor of cytokine signaling 2, cytotoxic T lymphocyte-associated protein 4 (CTLA4), selectin lymphocyte, interferon-γ (IFN-γ), programmed cell death 1 (Pdcd1), and granzyme B (Gzmb), was seen 8 days after transplantation along with histological evidence of severe allograft rejection...
April 2012: Liver Transplantation
P Souter, L K Bay, N Andreakis, N Császár, F O Seneca, M J H van Oppen
We report an accurate multiplex reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay, capable of reproducing gene expression profiles from 16 target genes [12 genes of interest (GOIs) and four reference genes (RGs)] in Acropora millepora, a common reef-building model coral species. The 12 GOIs have known or putative roles in the coral bleaching response, yet the method is not restricted to this particular assay and gene set. The procedure is based on the Beckman Coulter (Fullerton, CA, USA) GenomeLab™ GeXP Genetic Analysis System and bridges the gap between quantitative real-time PCR (qPCR) expression analysis of a single or a small number of genes and microarray gene expression surveys of thousands of genes...
March 2011: Molecular Ecology Resources
Janice E Drew, Claus-Dieter Mayer, Andrew J Farquharson, Pauline Young, Lawrence N Barrera
Colon cancers are characterized by aberrant gene expression signatures associated with disease initiation and progression. Identification of aberrant gene expression associated with colon carcinogenesis has increased significantly with application of gene array technologies. Downstream processing of these data has been hindered by the lack of robust multiplexed gene quantitative technologies facilitating study of the identified multiple gene targets. The GenomeLab Genetic Analysis System presents a novel technology platform for quantitative multiplexed gene expression analysis...
March 2011: Journal of Molecular Diagnostics: JMD
Ketli Ho, Latifah Saiful Yazan, Norsharina Ismail, Maznah Ismail
Vanillin is useful as anti-sickle cell anemia, anti-mutagen and anti-bacteria agent. However, vanillin must be administered at high concentration and cannot be oxidized by the upper gastrointestinal track of patients to be medically effective. In this study, we assessed the toxic effect of vanillin when administered in an un-oxidized form at high concentrations (150 and 300 mg/kg) via oral and intra-peritoneal injection. It was found that 300 mg/kg vanillin injection caused the rats to be unconscious without exerting any toxic effect on blood cells, kidney and liver...
January 2011: Food and Chemical Toxicology
Meng Qin, Da-yan Wang, Fang Huang, Kai Nie, Mei Qu, Miao Wang, Yue-long Shu, Xue-jun Ma
A novel application of the GeXP genetic analysis system for the differential detection of pandemic influenza A H1N1 from seasonal influenza A H1N1 and H3N2 is described. The assay was evaluated using identified influenza viruses and clinical samples. The results indicate that the assay is both highly sensitive and specific for the detection of the pandemic influenza A H1N1 virus with a detection limit of 10 copies per reaction superior to that of assays in use currently. The assay is able to detect potential mixed infections...
September 2010: Journal of Virological Methods
Maznah Ismail, Ghanya Al-Naqeeb, Wan Abd Aziz Bin Mamat, Zalinah Ahmad
BACKGROUND: Gamma-oryzanol (OR), a phytosteryl ferulate mixture extracted from rice bran oil, has a wide spectrum of biological activities in particular, it has antioxidant properties. METHODS: The regulatory effect of gamma-oryzanol rich fraction (ORF) extracted and fractionated from rice bran using supercritical fluid extraction (SFE) in comparison with commercially available OR on 14 antioxidant and oxidative stress related genes was determined in rat liver. Rats were subjected to a swimming exercise program for 10 weeks to induce stress and were further treated with either ORF at 125, 250 and 500 mg/kg or OR at 100 mg/kg in emulsion forms for the last 5 weeks of the swimming program being carried out...
2010: Nutrition & Metabolism
Maria A Nagel, Don Gilden, Ted Shade, Bifeng Gao, Randall J Cohrs
Varicella zoster virus (VZV) becomes latent in ganglionic neurons along the entire neuraxis. Although all predicted VZV open reading frames (ORFs) have been detected by macroarray and microarray analysis in virus-infected cells in culture where virus gene expression is abundant, array technology does not detect all VZV gene transcripts in latently infected human ganglia, where the abundance of ganglionic RNA is low and VZV gene transcription is highly variable. Using reverse transcription-polymerase chain reaction (RT-PCR) and the GenomeLab Genetic Analysis System (GeXPS), transcripts mapping to all 68 predicted unique VZV ORFs were detected in VZV-infected MeWo cells...
April 2009: Journal of Virological Methods
Alex J Rai, Rashmi M Kamath, William Gerald, Martin Fleisher
The GeXP genetic analysis system allows for multiplexed detection and quantitation of up to 35 genes in 192 samples in a single analysis. The analytical procedure includes modified reverse transcription and PCR amplification, followed by capillary electrophoretic separation. RNA material from multiple sample types can be used, including blood, cell lines, and tissue material. This instrumentation has a lower limit of detection of <2 ng with a dynamic range greater than two orders of magnitude, as tested in this validation...
March 2009: Analytical and Bioanalytical Chemistry
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