Expression of genes KCNQ1 and HERG encoding potassium ion channels Ikr, Iks in long QT syndrome

Background: The KCNQ1 and HERG genes mutations are responsible for specific types of congenital long QT syndrome (LQT). Aim: To examine the expression of KCNQ1 and HERG genes that encode potassium channels (rapid and slow) responsible for the occurrence of particular types of LQT syndrome. The study also attempted to prove that beta-actin is a good endogenous control when determining the expression of the studied genes. Methods: The study enrolled six families whose members suffered from either LQT1 or LQT2, or were healthy. Examination of gene expression was achieved with quantitative PCR (QRT-PCR). Expression of the investigated genes was inferred from the analysis of the number of mRNA copies per 1 mg total RNA isolated from whole blood. On the basis of KCNQ1 gene expression profile, the presence of, or absence of, LQT1 could be confirmed. Results and conclusions: The study revealed a statistically significant difference (p = 0.031) between the number of KCNQ1 gene copies in patients and healthy controls. On the basis of HERG (KCNH2) gene expression profile, patients with LQT2 cannot be unequivocally differentiated from healthy subjects (p = 0.37). Kardiol Pol 2011; 69, 5: 423–429Wstęp i cel: Głównym celem pracy było zbadanie ekspresji genów HERG i KCNQ1, kodujących kanały potasowe (szybkie i wolne), odpowiadających za wystąpienie określonego rodzaju zespołu długiego QT (LQTS). Metody: Do badania włączono 6 rodzin, u członków których zdiagnozowano LQTS1 lub LQTS2, lub zdrowych. Badanie miało na celu udowodnienie, że beta-aktyna stanowi dobrą kontrolę endogenną przy ustalaniu ekspresji badanych genów. Do badania ekspresji genów wykorzystano ilościową analizę PCR w czasie rzeczywistym (QRT-PCR). Ekspresję badanych genów przedstawiono jako liczbę kopii mRNA w przeliczeniu na 1 mg całkowitego RNA izolowanego z krwi pełnej. Dane zostały wyeksportowane z arkusza Excel do programu analizy danych Statistica V.7.1. Wyniki i wnioski: Na podstawie profilu ekspresji genu KCNQ1 można potwierdzić występowanie zespołu LQTS1. Badania wykazały statystycznie istotną różnicę (p = 0,031) między liczbą KCNQ1 kopii genu u osób chorych i zdrowych. Na podstawie profilu ekspresji genu HERG (KCNH2) chorych z LQTS2 nie można jednoznacznie odróżnić od osób zdrowych (p = 0,37). Kardiol Pol 2011; 69, 5: 423–42

Aspirin – 115 years after the discovery

Aspirin has been known as a commercial drug for over a century, however, a much deeper understanding of its mechanism of action as an inhibitor of cyclooxygenase (COX) activity and thus, of prostanoid synthesis, is still lacking. Recent advances in understanding the central role of platelets in the pathophysiology of cardiovascular diseases and the identification of novel lipid mediators synthesized in the presence of aspirin have increased research upon the mechanisms of aspirin action.Aspiryna jest lekiem dostępnym komercyjnie od ponad stu lat, chociaż wciąż brakuje głębszego zrozumienia mechanizmu jej działania jako inhibitora aktywności cyklooksygenazy i syntezy prostanoidów. Niedawne odkrycia dotyczące centralnej roli płytek krwi w patofizjologii chorób układu sercowo-naczyniowego oraz identyfikacja nowych mediatorów lipidowych syntetyzowanych w obecności aspiryny nasiliły badania nad mechanizmami działania aspiryny

Genetical polymporphism of chosen CYP2D6 alleles among the patients with dilated cardiomyopathy and myocarditis

Cytochrome P450 2D6 has been reported to possess variation in the encoding gene that aff ects enzymatic activity. Interindividual diff erences in CYP2D6 activity can produce adverse eff ects or lack of therapeutic eff ect under standard therapy. The aim of the study was to genotyope the chosen alleles of CYP2D6 among the patients with clinically confi rmed myocarditis or dilated cardiomyopathy. Tetra-primer PCR assays was used to detect mutations in CYP2D6 *3, *4 and *6 allelles among 53 patients. A multiplex long PCR was used to genotype CYP2D6*5 allele. Analysis showed the presence of one heterozygote for CYP2D6*3, two heterozygote for CYP2D6*6, ten heterozygote and two homozygote for CYP2D6*4 among the examined patients. One person with deletion of CYP2D6 locus was also detected. Presented methods will facilitate and accelerate the detailed pharmacogenomic analysis of CYP2D6.Cytochrom CYP2D6 ma różne warianty sekwencji kodującego go genu, wpływające na jego aktywność enzymatyczną. Międzyosobnicze różnice w aktywności izoenzymu CYP2D6 mogą prowadzić do działań niepożądanych lub braku skuteczności terapeutycznej standardowych dawek leku. Celem pracy było genotypowanie wybranych alleli CYP2D6 w grupie pacjentów z potwierdzonym klinicznie stanem zapalnym mięśnia sercowego lub kardiomiopatią rozstrzeniową. W pracy zastosowano reakcję PCR z użyciem czterech starterów w celu detekcji alleli 3*,4* i 6* CYP2D6 w grupie 53 pacjentów. Do wykrycia allelu CY2D6*5 zastosowano długołańcuchową reakcję PCR typu multipleks. Przeprowadzona analiza wykazała w badanej grupie pacjentów obecność jednej heterozygoty dla allelu CYP2D6*3, dwóch heterozygot dla allelu CYP2D*6, dziesięciu heterozygot i dwóch homozygot dla alellu CYP2D6*4. Znaleziono także jedną osobę z delecją locus CYP2D6. Przedstawione metody ułatwiają i przyspieszają dokładną analizę farmakogenetyczną izoformy CYP2D6

In vitro and in silico study on the effect of carvedilol and sorafenib alone and in combination on the growth and inflammatory response of melanoma cells

Melanoma is an aggressive skin cancer. Increasing evidence has shown the role of β-adrenergic receptors in the pathogenesis of melanoma. Carvedilol is a widely used non-selective β-AR antagonist with potential anticancer activity. The purpose of the study was to estimate the influence of carvedilol and sorafenib alone and in combination on the growth and inflammatory response of C32 and A2058 melanoma cells. Furthermore, this study also aimed to predict the probable interaction of carvedilol and sorafenib when administered together. Predictive study of the interaction of carvedilol and sorafenib was performed using the ChemDIS-Mixture system. Carvedilol and sorafenib alone and in combination showed a growth inhibitory effect on cells. The greatest synergistic antiproliferative effect on both cell lines was observed at Car 5 μM combined with Sor 5 μM. Analysis in silico identified diseases, proteins, and metabolic pathways that can be affected by the interaction of carvedilol and sorafenib. The results obtained demonstrated that carvedilol and sorafenib modulated the secretion of IL-8 by IL-1β-stimulated by melanoma cell lines but the use of a combination of both drugs did not intensify the effect. In summary, the results presented indicate that the combination of carvedilol and sorafenib may have a promising anticancer effect on melanoma cells

Mechanism of Pterostilbene-Induced Cell Death in HT-29 Colon Cancer Cells

Pterostilbene is a dietary phytochemical that has been found to possess several biological activities, such as antioxidant and anti-inflammatory. Recent studies have shown that it exhibits the hallmark characteristics of an anticancer agent. The aim of the study was to investigate the anticancer activity of pterostilbene against HT-29 human colon cancer cells, focusing on its influence on cell growth, differentiation, and the ability of this stilbene to induce cell death. To clarify the mechanism of pterostilbene activity against colon cancer cells, changes in the expression of several genes and proteins that are directly related to cell proliferation, signal transduction pathways, apoptosis, and autophagy were also evaluated. Cell growth and proliferation of cells exposed to pterostilbene (5–100 µM) were determined by SRB and BRDU assays. Flow cytometric analyses were used for cell cycle progression. Further molecular investigations were performed using quantitative real-time RT-PCR. The expression of the signaling proteins studied was determined by the ELISA method. The results revealed that pterostilbene inhibited proliferation and induced the death of HT-29 colon cancer cells. Pterostilbene, depending on concentration, caused inhibition of proliferation, G1 cell arrest, and/or triggered apoptosis in HT-29 cells. These effects were mediated by the down-regulation of the STAT3 and AKT kinase pathways. It may be concluded that pterostilbene could be considered as a potential therapeutic option in the treatment of colon cancer in the future

Chorzy trudni nietypowiPodłoże genetyczne rodzinnej kardiomiopatii przerostowej w czteropokoleniowej rodzinie – opis przypadku

Genetic profile of four-generation family with hypertrophic cardiomyopathy is presented. The alterations in the MYH7 gene sequences were identified. Genetic background of familial hypertrophic cardiomyopathy is reviewed and discussed

Familial hypertrophic cardiomyopathy. Insertion-deletion polymorphism of angiotensin-converting enzyme and angiotensin II receptor

Background: Hypertrophic cardiomyopathy (HCM) is a genetic-based disease. Several gene mutations leading to HCM development have been described.Aim: Detailed examination of phenotype and genotype of a family with HCM.Methods: Clinical and genetic examinations were performed in a family with HCM, in which 3 sick persons with different disease phenotype were found.Results: In all sick persons the same molecular substitution G->A (AGG->AAG) was noticed. It led to substitution Arg780-Lys in exon 21 β-myosin heavy chain gene, which was responsible for the development of the disease. Insertion- deletion polymorphism analysis in ACE gene revealed D/D (deletion/deletion) genotype in proband and D/I (deletion/ insertion) phenotype in his mother and sister, who were heterozygous. Polymorphism A1166C analysis in AT1 gene revealed the presence of genotype A/A in proband and A/C in his mother and sister. In proband and his sister a very similar phenotype was observed, whereas they had different polymorphism for ACE gene and angiotensin 1 receptor gene. In sick proband's mother, who had phenotype different to her children, the same polymorphism as in his daughter was noticed.Conclusions: In the described family with HCM, different phenotype and polymorphism of ACE and AT1 genes were found