ABSTRACT: Congestive heart failure was done by artificial myocardial infarction. After extracting total RNA from control and infarcted ventricles with Triazol, messenger RNA was isolated with Qiagen mRNA isolation kit. About 1.5ug poly A+ RNA was taken for probe preparation. Probe was labeled with Alexa Fluor 546 and 657. Labeled probes were hybridized with Qiagen rat unigene oligo library. After 2 low and 1 high stringency washes Prolong Antifade was added to the slides to prevent bleaching effect. The data ratio was normalized using a location and intensity dependent Lowess formula. Keywords: other
Project description:Congestive heart failure was done by artificial myocardial infarction. After extracting total RNA from control and infarcted ventricles with Triazol, messenger RNA was isolated with Qiagen mRNA isolation kit. About 1.5ug poly A+ RNA was taken for probe preparation. Probe was labeled with Alexa Fluor 546 and 657. Labeled probes were hybridized with Qiagen rat unigene oligo library. After 2 low and 1 high stringency washes Prolong Antifade was added to the slides to prevent bleaching effect. The data ratio was normalized using a location and intensity dependent Lowess formula.
Project description:Rat ventricular cardiomyocytes were prepared from 18 day old fetus. Approximately 6 million cells were plated on 100 mm culture plate. The serum used in the medium was depleted from thyroid hormone by passing through anion exchange resin column. After 48 hours of growth with 70% confluency, cells were exposed to either T3, DITPA or CGS for another 48 hours before harvesting. Final concentrations of T3, DITPA and CGS in the medium were 6nM, 218nM and 1nM respectively. RNA was prepared from cells with Triazol. Twenty micrograms of total RNA was reverse transcribed and labeled with Alexa Fluor 546 and 647. Labeled probes were hybridized with Qiagen's rat unigene oligo library. After 2 low and 1 high stringency washes Prolong antifade was added to the slides to prevent bleaching effect. Slides were scanned in Array Worx scanner with dual wave length. The data ratio was normalized using a location and intensity dependent Lowess formula. Keywords: equivalent probe
Project description:Molecular analysis of the effect left ventricular assist device (LVAD) support has on congestive heart failure patients. Keywords = Congestive heart failure, left ventricular assist device, eNOS, gene, dimethylarginine dimethylaminohydrolase Keywords: other
Project description:Three weeks after infarction, age matched rats with large infarction by ECG criteria were randomly assigned to one of the three groups: 1) Treatment with Captopril for 21 days and a daily subcutaneous dose of DITPA for the last 10 days of treatment, 2) Treatment for 10 days with DITPA alone, 3) Control heart failure and 4) Sham-operated. Keywords: Drug Effects on Heart Failure Total RNA was prepared from left ventricle using TRIzol and later Poly A+ RNA was isolated with Oligotex beads. About 0.5ug Poly A+ RNA was reverse transcribed and labeled with Alexa Fluor 546 and 647. Labeled probed were hybridized with Operon's rat Unigene oligo library printed on glass slides. After 2 lows and 1 high stringency washes, Prolong Antifade was added to the slides to prevent any bleaching effect. The experiments were repeated 2-4 times by dye-swaping. Slides were scanned in Array Worx scanner with dual wave length. The data ratio was normalized using a location and intensity dependent Lowess formula.
Project description:Canine tachycardia-induced cardiomyopathy caused by several weeks of rapid ventricular pacing is a well-established animal model of congestive heart failure. However, little is known about the underlying changes in gene expression that occur in the canine myocardium after the induction of heart failure. This project aims to compare expression profiles in left ventricular free wall samples from control dogs and dogs with pacing-induced heart failure on the custom MuscleChip. Keywords: other
Project description:Mice were intranasally instilled at 6 weeks of age with silica, titanium dioxide, or saline control and macrophages were harvested by broncho-alveolar lavage at 14 weeks of age. RNA was isolated using RNeasy Mini kits (Qiagen). Due to low yield of RNA, mRNA was amplifed by RiboAmp kits (Arcturus) and resulting aRNA was indirectly labeled with Alexa-Fluor 647 dye (Molecular Probes). This was hybridized against Universal Mouse Reference RNA (Stratagene) which was indirectly labeled with Alexa-Fluor 555 dye (Molecular Probes). Slides were printed in house using PCR product cDNA targets on GAPS slides (Corning). Slides were scanned with an Axon GenePix 4000B scanner and data was analyzed using GenePix Pro 4.0 software and Iobion GeneTraffic Duo software. Data was normalized to a total 1:1 ratio of read to green dye. Keywords: parallel sample
Project description:In this study, we aimed to identify novel serum biomarkers in feline primary cardiomyopathies with congestive heart failure, which can help to understand disease pathogenesis and assist the disease diagnosis, prognosis and management. The study included 15 cats diagnosed with CHF and CM, 5 cats with asymptomatic CM and 15 healthy cats.
Project description:Streptococcus pneumoniae (the pneumococcus) account for significant morbidity and mortality worldwide, causing life-threatening diseases such as pneumonia, bacteremia and meningitis. In this study, we used microarray analysis to compare gene expression patterns of either serotype 4 or serotype 6A pneumococci in the nasopharynx and blood of mice, as a model to identify genes involved in invasion of blood in the context of occult bacteremia in humans. Microarray experiments were performed on whole genome S. pneumoniae PCR arrays obtained from the Bacterial Microarray Group at St George's Hospital Medical School, London (http://bugs.sghms.ac.uk/). The array was designed using TIGR4 base strain annotation and extra target genes from strain R6. Pair-wise comparisons were made between the nasopharynx and blood RNA samples labeled with either Alexa Fluor 546 or Alexa Fluor 647 dye from the 48, 72 and 96 h time points.
Project description:Oxidative stress is recognazed as a hallmark of cancer. However, little is known about intratumor heterogeneity of reactive oxygen species (ROS) and its influence on tumor behavior, largely due to technical challenges inherent in detecting intratumor ROS at single-cell resolution. We have developed a tumor-targeted H2O2 probe (T-AP1) by conjugating trastuzumab with both Alexa Fluor 647 (AF647), an H2O2-insensitive fluorophore, and peroxy green 1 (PG1), an irreversible H2O2-sensitive chemical probe to provide a “ratiometric” readout, allowing H2O2 measurements independent of accumulated T-AP1 levels. Given the ability of T-AP1 to persistently mark tumor cells that experience H2O2 exposure, we employed cell sorting to isolate tumor cells with high (PG1highAF647+) and low (PG1lowAF647+) H2O2 exposure levels from T-AP1–labeled NCI-N87 xenograft tumors. PG1highAF647+ and PG1lowAF647+ cells obtained via cell sorting from three biologically independent N87 tumors were subjected to RNA sequencing.