Ischemic postconditioning alters the gene expression pattern of the ischemic heart.

To profile changes in gene expression in response to ischemic postconditioning, isolated rat hearts were subjected to 30 min of regional ischemia followed by 120 min of reperfusion with or without postconditioning. At the end of reperfusion, cardiac RNA was assayed by DNA microarrays (31,000 format), verified by quantitative real-time polymerase chain reaction (QRT-PCR). Postconditioning significantly up-regulated 50 genes and down-regulated 58 different genes, including pyruvate dehydrogenase, 60 kDa heat shock protein 1, lipoprotein lipase, gamma-sarcoglycan, and phospholipase C.

Altered expression of genes for Kir ion channels in dilated cardiomyopathy.

Dilated cardiomyopathy (DCM) is a multifactorial disease characterized by left ventricular dilation that is associated with systolic dysfunction and increased action potential duration. The Kir2.x K⁺ channels (encoded by KCNJ genes) regulate the inward rectifier current (IK1) contributing to the final repolarization in cardiac muscle.

Role of iNOS and peroxynitrite-matrix metalloproteinase-2 signaling in myocardial late preconditioning in rats.

We have previously shown that the inhibition of myocardial nitric oxide (NO) and peroxynitrite-matrix metalloproteinase (MMP) signaling by early preconditioning (PC) is involved in its cardioprotective effect. Therefore, in the present study, we investigated the role of NO and peroxynitrite-MMP signaling in the development of late PC. PC was performed by five consecutive cycles of 4-min coronary occlusion and 4-min reperfusion in anesthetized rats in vivo.

Expression of genes related to oxidative/nitrosative stress in mouse hearts: effect of preconditioning and cholesterol diet.

Background: The aim of our study was to explore the effect of high-cholesterol diet and preconditioning on cardiac gene expression patterns in mouse hearts, focusing on genes involved in nitric oxide (NO) and free radical signaling and the mevalonate pathway.

Material/methods: Mice were fed 2% high-cholesterol or normal diet for 8 weeks. Hearts isolated from both groups were subjected to either a preconditioning (PC) protocol (3 cycles of 5 min ischemia and 5 min aerobic perfusion) or a time-matched non-preconditioning protocol followed by 30 min global test ischemia and 2 hour reperfusion.

Gene and protein expression changes in response to normoxic perfusion in mouse hearts.

Introduction: Although crystalloid-perfused isolated heart models are widely used in cardiovascular research, there are several limitations of these techniques. Changes in cardiac gene expression pattern due to normoxic perfusion itself have not been studied, despite its potential importance to provide useful information on limitations of this model. Therefore, here we investigated the time-dependent effect of normoxic, normothermic perfusion on global gene expression at mRNA and protein levels.

Multiplex site-directed mutagenesis strategy including high-efficiency selection of the mutant PCR products.

Site-directed mutagenesis is of great importance for probing the structure/function relationship of proteins. Developing our previous method (Nagy et al. Anal Biochem 324:301-303, 2004), here we report a multiplex strategy for site-directed mutagenesis using PCR in one tube to introduce a single mutation into three or more genes at the same time.

Improved DOP-PCR-based representational whole-genome amplification using quantitative real-time PCR.

In many cases, only a minute amount of partially degraded genomic DNA can be extracted from archived clinical samples. Diverse whole-genome amplification methods are applied to provide sufficient amount of DNA for comparative genome hybridization, single-nucleotide polymorphism, and microsatellite analyses. In these applications, the reliability of the amplification techniques is particularly important.