Comparison of Cardioplegic Solutions in Coronary Bypass Surgery Over Autophagy and Apoptosis Mechanisms. / Comparação de Soluções Cardioplégicas em Cirurgia de Revascularização Miocárdica sobre Mecanismos de Autofagia e Apoptose.

BACKGROUND: Coronary artery disease (CAD) due to myocardial ischemia causes permanent loss of heart tissue. OBJECTIVES: We aimed to demonstrate the possible damage to the myocardium at the molecular level through the mechanisms of autophagy and apoptosis in coronary bypass surgery patients. METHODS: One group was administered a Custodiol cardioplegia solution, and the other group was administered a Blood cardioplegia solution. Two myocardial samples were collected from each patient during the operation, just before cardiac arrest and after the aortic cross-clamp was released. The expressions of autophagy and apoptosis markers were evaluated. The level of statistical significance adopted was 5%. RESULTS: The expression of the BECLIN gene was significant in the myocardial tissues in the BC group (p=0.0078). CASPASE 3, 8, and 9 gene expression levels were significantly lower in the CC group. Postoperative TnT levels were significantly different between the groups (p=0.0072). CASPASE 8 and CASPASE 9 gene expressions were similar before and after aortic cross-clamping (p=0.8552, p=0.8891). In the CC group, CASPASE 3, CASPASE 8, and CASPASE 9 gene expression levels were not found to be significantly different in tissue samples taken after aortic cross-clamping (p=0.7354, p=0.0758, p=0.4128, respectively). CONCLUSIONS: With our findings, we believe that CC and BC solutions do not have a significant difference in terms of myocardial protection during bypass operations.

FUNDAMENTO: A doença arterial coronariana (DAC) devido à isquemia miocárdica causa perda permanente de tecido cardíaco. OBJETIVOS: Nosso objetivo foi demonstrar o possível dano ao miocárdio em nível molecular através dos mecanismos de autofagia e apoptose em pacientes submetidos à cirurgia de revascularização miocárdica. MÉTODOS: Um grupo recebeu uma solução de cardioplegia Custodiol e o outro grupo uma solução de cardioplegia sanguínea. Duas amostras miocárdicas foram coletadas de cada paciente durante a operação, imediatamente antes da parada cardíaca e após a liberação do pinçamento aórtico. Foram avaliadas as expressões de marcadores de autofagia e apoptose. O nível de significância estatística adotado foi de 5%. RESULTADOS: A expressão do gene BECLIN foi significativa nos tecidos miocárdicos do grupo CS (p=0,0078). Os níveis de expressão dos genes CASPASE 3, 8 e 9 foram significativamente menores no grupo CC. Os níveis pós-operatórios de TnT foram significativamente diferentes entre os grupos (p=0,0072). As expressões dos genes CASPASE 8 e CASPASE 9 foram semelhantes antes e depois do pinçamento aórtico (p=0,8552, p=0,8891). No grupo CC, os níveis de expressão gênica de CASPASE 3, CASPASE 8 e CASPASE 9 não foram significativamente diferentes em amostras de tecido coletadas após pinçamento aórtico (p=0,7354, p=0,0758, p=0,4128, respectivamente). CONCLUSÕES: Com nossos achados, acreditamos que as soluções CC e CS não apresentam diferença significativa em termos de proteção miocárdica durante as operações de by-pass.

Overexpression of the PTEN Gene in Myocardial Tissues of Coronary Bypass Surgery Patients. / Superexpressão Gênica PTEN em Tecidos Miocárdicos de Pacientes de Cirurgia de Revascularização Miocárdica.

BACKGROUND: Coronary artery disease is a complex disorder that causes death worldwide. One of the genes involved in developing this disease may be PTEN. OBJECTIVES: This study aimed to investigate the PTEN gene and protein expression in tissue and blood samples taken from coronary bypass surgery patients. METHODS: Molecular studies were performed at Erciyes University Genome and Stem Cell Center (GENKOK). Right atrial appendage and blood samples were taken from the central vein of 22 coronary bypass surgery patients before starting and ending cardiopulmonary bypass. PTEN expression was determined using quantitative real-time PCR and western blot analysis. The significance level was accepted as p<0.05. RESULTS: There was no significant difference in the PTEN gene expression in blood samples taken before and after cardiopulmonary bypass. However, a substantial increase in both protein and gene expression levels of P-PTEN and PTEN was observed in the tissue samples. Myocardial expression of the PTEN gene was significantly increased at the end of the cardiopulmonary bypass. PTEN gene expression in the post-cardiopulmonary bypass period was increased when compared to the pre-bypass period, but it was insignificant when compared to healthy controls. CONCLUSION: This study first revealed the role of the PTEN gene by analyzing both mRNA and protein expression in coronary bypass patients, appearing in both myocardial tissue and blood samples. Increased levels of PTEN may be a marker in myocardial tissue for patients with coronary artery disease.

FUNDAMENTO: A doença arterial coronariana é um distúrbio complexo que causa morte em todo o mundo. Um dos genes envolvidos no desenvolvimento dessa doença pode ser o PTEN. OBJETIVOS: Nosso objetivo foi investigar a expressão gênica e proteica do PTEN em amostras de tecido e sangue retiradas de pacientes submetidos à cirurgia de revascularização miocárdica. MÉTODOS: Foram realizados estudos moleculares no Centro de estudos do genoma humano e células-tronco da Universidade Erciyes (GENKOK). Amostras do apêndice atrial direito e de sangue foram coletadas da veia central de 22 pacientes submetidos à cirurgia de revascularização miocárdica antes de iniciar e terminar a circulação extracorpórea. A expressão do PTEN foi determinada usando PCR quantitativo em tempo real e análise de Western Blot. O nível de significância aceito foi de p<0,05. RESULTADOS: Não houve diferença significativa na expressão gênica do PTEN em amostras de sangue coletadas antes e depois da circulação extracorpórea. Entretanto, foi observado um aumento substancial nos níveis de expressão gênica e proteica de P-PTEN e PTEN nas amostras de tecido. A expressão gênica miocárdica PTEN aumentou significativamente ao final da circulação extracorpórea. A expressão gênica do PTEN no período pós-circulação extracorpórea aumentou em comparação com o período pré-circulação extracorpórea, mas não foi um aumento significativo em comparação com sujeitos saudáveis do grupo de controle. CONCLUSÃO: Este estudo revelou pela primeira vez o papel do gene PTEN analisando a expressão de mRNA e de proteína em pacientes com revascularização miocárdica, que se manifesta tanto em tecido miocárdico quanto em amostras de sangue. O aumento dos níveis de PTEN pode ser um marcador no tecido miocárdico para pacientes com doença arterial coronariana.

Comparação de Soluções Cardioplégicas em Cirurgia de Revascularização Miocárdica sobre Mecanismos de Autofagia e Apoptose / Comparison of Cardioplegic Solutions in Coronary Bypass Surgery Over Autophagy and Apoptosis Mechanisms

Resumo Fundamento A doença arterial coronariana (DAC) devido à isquemia miocárdica causa perda permanente de tecido cardíaco. Objetivos Nosso objetivo foi demonstrar o possível dano ao miocárdio em nível molecular através dos mecanismos de autofagia e apoptose em pacientes submetidos à cirurgia de revascularização miocárdica. Métodos Um grupo recebeu uma solução de cardioplegia Custodiol e o outro grupo uma solução de cardioplegia sanguínea. Duas amostras miocárdicas foram coletadas de cada paciente durante a operação, imediatamente antes da parada cardíaca e após a liberação do pinçamento aórtico. Foram avaliadas as expressões de marcadores de autofagia e apoptose. O nível de significância estatística adotado foi de 5%. Resultados A expressão do gene BECLIN foi significativa nos tecidos miocárdicos do grupo CS (p=0,0078). Os níveis de expressão dos genes CASPASE 3, 8 e 9 foram significativamente menores no grupo CC. Os níveis pós-operatórios de TnT foram significativamente diferentes entre os grupos (p=0,0072). As expressões dos genes CASPASE 8 e CASPASE 9 foram semelhantes antes e depois do pinçamento aórtico (p=0,8552, p=0,8891). No grupo CC, os níveis de expressão gênica de CASPASE 3, CASPASE 8 e CASPASE 9 não foram significativamente diferentes em amostras de tecido coletadas após pinçamento aórtico (p=0,7354, p=0,0758, p=0,4128, respectivamente). Conclusões Com nossos achados, acreditamos que as soluções CC e CS não apresentam diferença significativa em termos de proteção miocárdica durante as operações de by-pass.

Abstract Background Coronary artery disease (CAD) due to myocardial ischemia causes permanent loss of heart tissue. Objectives We aimed to demonstrate the possible damage to the myocardium at the molecular level through the mechanisms of autophagy and apoptosis in coronary bypass surgery patients. Methods One group was administered a Custodiol cardioplegia solution, and the other group was administered a Blood cardioplegia solution. Two myocardial samples were collected from each patient during the operation, just before cardiac arrest and after the aortic cross-clamp was released. The expressions of autophagy and apoptosis markers were evaluated. The level of statistical significance adopted was 5%. Results The expression of the BECLIN gene was significant in the myocardial tissues in the BC group (p=0.0078). CASPASE 3, 8, and 9 gene expression levels were significantly lower in the CC group. Postoperative TnT levels were significantly different between the groups (p=0.0072). CASPASE 8 and CASPASE 9 gene expressions were similar before and after aortic cross-clamping (p=0.8552, p=0.8891). In the CC group, CASPASE 3, CASPASE 8, and CASPASE 9 gene expression levels were not found to be significantly different in tissue samples taken after aortic cross-clamping (p=0.7354, p=0.0758, p=0.4128, respectively). Conclusions With our findings, we believe that CC and BC solutions do not have a significant difference in terms of myocardial protection during bypass operations.

Superexpressão Gênica PTEN em Tecidos Miocárdicos de Pacientes de Cirurgia de Revascularização Miocárdica / Overexpression of the PTEN Gene in Myocardial Tissues of Coronary Bypass Surgery Patients

Resumo Fundamento: A doença arterial coronariana é um distúrbio complexo que causa morte em todo o mundo. Um dos genes envolvidos no desenvolvimento dessa doença pode ser o PTEN. Objetivos: Nosso objetivo foi investigar a expressão gênica e proteica do PTEN em amostras de tecido e sangue retiradas de pacientes submetidos à cirurgia de revascularização miocárdica. Métodos: Foram realizados estudos moleculares no Centro de estudos do genoma humano e células-tronco da Universidade Erciyes (GENKOK). Amostras do apêndice atrial direito e de sangue foram coletadas da veia central de 22 pacientes submetidos à cirurgia de revascularização miocárdica antes de iniciar e terminar a circulação extracorpórea. A expressão do PTEN foi determinada usando PCR quantitativo em tempo real e análise de Western Blot. O nível de significância aceito foi de p<0,05. Resultados: Não houve diferença significativa na expressão gênica do PTEN em amostras de sangue coletadas antes e depois da circulação extracorpórea. Entretanto, foi observado um aumento substancial nos níveis de expressão gênica e proteica de P-PTEN e PTEN nas amostras de tecido. A expressão gênica miocárdica PTEN aumentou significativamente ao final da circulação extracorpórea. A expressão gênica do PTEN no período pós-circulação extracorpórea aumentou em comparação com o período pré-circulação extracorpórea, mas não foi um aumento significativo em comparação com sujeitos saudáveis do grupo de controle. Conclusão: Este estudo revelou pela primeira vez o papel do gene PTEN analisando a expressão de mRNA e de proteína em pacientes com revascularização miocárdica, que se manifesta tanto em tecido miocárdico quanto em amostras de sangue. O aumento dos níveis de PTEN pode ser um marcador no tecido miocárdico para pacientes com doença arterial coronariana.

Abstract Background: Coronary artery disease is a complex disorder that causes death worldwide. One of the genes involved in developing this disease may be PTEN. Objectives: This study aimed to investigate the PTEN gene and protein expression in tissue and blood samples taken from coronary bypass surgery patients. Methods: Molecular studies were performed at Erciyes University Genome and Stem Cell Center (GENKOK). Right atrial appendage and blood samples were taken from the central vein of 22 coronary bypass surgery patients before starting and ending cardiopulmonary bypass. PTEN expression was determined using quantitative real-time PCR and western blot analysis. The significance level was accepted as p<0.05. Results: There was no significant difference in the PTEN gene expression in blood samples taken before and after cardiopulmonary bypass. However, a substantial increase in both protein and gene expression levels of P-PTEN and PTEN was observed in the tissue samples. Myocardial expression of the PTEN gene was significantly increased at the end of the cardiopulmonary bypass. PTEN gene expression in the post-cardiopulmonary bypass period was increased when compared to the pre-bypass period, but it was insignificant when compared to healthy controls. Conclusion: This study first revealed the role of the PTEN gene by analyzing both mRNA and protein expression in coronary bypass patients, appearing in both myocardial tissue and blood samples. Increased levels of PTEN may be a marker in myocardial tissue for patients with coronary artery disease.

Blood mRNA Expression Profiles of Autophagy, Apoptosis, and Hypoxia Markers on Blood Cardioplegia and Custodiol Cardioplegia Groups

Abstract Introduction: Blood cardioplegia (BC) and Custodiol cardioplegia (CC) have been used for a long time in open heart surgery and are highly effective solutions. The most controversial issue among these two is whether there is any difference between them regarding myocardial damage after ischemia surgery. In this study, autophagy, apoptosis, and hypoxia markers were investigated and that way we evaluated the differences between BC and CC patients. Methods: A total of 30 patients were included in this study, using two different cardioplegic solutions. Three different whole blood samples of the patients were taken from a central vein (preoperatively, immediately postoperatively, and one day after surgery). Total ribonucleic acid was extracted from these samples. Quantitative real-time polymerase chain reaction was performed, and changes in gene expression were determined by the 2-∆∆Ct method of relative quantification. Results: In the CC group, Beclin gene expression level was found to be higher and this difference was statistically significant (P=0.0024). Similarly, cysteine-aspartic acid protease (caspase) 9 and hypoxia-inducible factor 1α messenger ribonucleic acid (mRNA) gene expression level increased and were significantly different in the CC group. In the BC group, Beclin and microtubule-associated protein light chain 3 expressions were higher in the samples taken one day after surgery. Caspases 3 and 8 gene expressions were significantly different in the BC group. Conclusion: As a result of the analysis performed between the two cardioplegia groups, it has been shown that CC harms the myocardium more than BC at the level of mRNA expression of related markers.

Blood mRNA Expression Profiles of Autophagy, Apoptosis, and Hypoxia Markers on Blood Cardioplegia and Custodiol Cardioplegia Groups.

INTRODUCTION: Blood cardioplegia (BC) and Custodiol cardioplegia (CC) have been used for a long time in open heart surgery and are highly effective solutions. The most controversial issue among these two is whether there is any difference between them regarding myocardial damage after ischemia surgery. In this study, autophagy, apoptosis, and hypoxia markers were investigated and that way we evaluated the differences between BC and CC patients. METHODS: A total of 30 patients were included in this study, using two different cardioplegic solutions. Three different whole blood samples of the patients were taken from a central vein (preoperatively, immediately postoperatively, and one day after surgery). Total ribonucleic acid was extracted from these samples. Quantitative real-time polymerase chain reaction was performed, and changes in gene expression were determined by the 2-∆∆Ct method of relative quantification. RESULTS: In the CC group, Beclin gene expression level was found to be higher and this difference was statistically significant (P=0.0024). Similarly, cysteine-aspartic acid protease (caspase) 9 and hypoxia-inducible factor 1α messenger ribonucleic acid (mRNA) gene expression level increased and were significantly different in the CC group. In the BC group, Beclin and microtubule-associated protein light chain 3 expressions were higher in the samples taken one day after surgery. Caspases 3 and 8 gene expressions were significantly different in the BC group. CONCLUSION: As a result of the analysis performed between the two cardioplegia groups, it has been shown that CC harms the myocardium more than BC at the level of mRNA expression of related markers.

Relationship of genetic factors with development of aortic dissection and aneurysm.

BACKGROUND: This study aims to investigate the relationship between the development of aortic dissections and aneurysms with the polymorphisms of angiotensin converting enzyme gene, methylenetetrahydrofolate reductase gene, plasminogen activator inhibitor-1 gene, and nitric oxide synthase gene. METHODS: Between April 2009 and July 2014, 38 patients with aortic dissections (28 males, 10 females; mean age 55.1±10.7 years; range, 30 to 78 years) and 67 patients with aortic aneurysms (57 males, 10 females; mean age 63.0±11.4 years; range, 31 to 82 years) were included in this cross-sectional study. The control group consisted of 60 healthy volunteers (41 males, 19 females; mean age 56.3±11.2 years; range, 30 to 82 years) without an aortic aneurysm or dissection, as assessed by thoracoabdominal computed tomography. The prespecified four genes were genotyped with competitive allelespecific polymerase chain reaction. RESULTS: The aortic dissection group had higher nitric oxide synthase-3 (4b/4b) expression levels, compared to the control group. The aortic aneurysm group had also higher nitric oxide synthase-3 (4b/4a) expression levels, compared to the control group. Compared to the control group, a higher rate of angiotensin converting enzyme I/D gene polymorphism was detected in the aneurysm group, while higher D/D polymorphism rates were found in the dissection group; although not statistically significant. CONCLUSION: Our study results suggest that the nitric oxide synthase-3 intron 4b/4b and nitric oxide synthase-3 intron 4b/4a gene polymorphisms can be used as a predictor of aortic dissection and aneurysm development.

The role of endothelial nitric oxide synthase gene G894T and intron 4 VNTR polymorphisms in hemodialysis patients with vascular access thrombosis.

OBJECTIVE: Endothelial nitric oxide synthase (eNOS) gene is a candidate gene in cardiovascular and renal diseases. Several polymorphic variations have been identified in eNOS gene. We investigated a potential role of arteriovenous fistula (AVF) thrombosis and intron 4 and G894T polymorphisms in chronic renal failure. METHODS: We performed a case-control observational study involving 79 with/without AVF thrombosis in chronic renal failure patients. All subjects were genotyped by the polymerase chain reaction (PCR) and PCR-Restriction Fragment Length Polymorphism. Genotype distribution and allele frequencies were compared between groups using the chi-square test. RESULTS: Genotype frequencies in patients with thrombosis were not significantly different from those of patients without thrombosis for eNOS G894T polymorphism (p=0.1). eNOS gene intron 4 a allele distributions seems to be associated with thrombosis in the groups. CONCLUSION: This study revealed that there was an association between eNOS intron 4 polymorphism and thrombosis in chronic renal failure patients. This data will be helpful in planning further eNOS association studies in vascular access thrombosis.

Association of TNF-α -308 G > A and ACE I/D gene polymorphisms in hemodialysis patients with arteriovenous fistula thrombosis.

PURPOSE: Tumor necrosis factor-α (TNF - α) -308 G > A promoter polymorphism seems to be associated with adverse clinical outcome in hemodialysis patients (HD). Angiotensin-converting enzyme (ACE) gene may be the causative factor contributing to the deterioration of renal functions. The aim of this study was to investigate the relationship between vascular access failure and the genetic polymorphisms of ACE and TNF-α gene. METHODS: We enrolled and genotyped 47 HD patients with arteriovenous fistula (AVF) thrombosis, 51 HD patients without AVF thrombosis, and 40 healthy controls. The genotypes of these polymorphisms were determined by polymerase chain reaction and restriction fragment length polymorphism. RESULTS: The genotype distribution of TNF-α -308 G > A in patients with thrombosis was significantly different from the patients without thrombosis (p = 0.008). There was no significant difference between the two groups in terms of ACE I/D polymorphism (p = 0.213). CONCLUSION: Our results propose that TNF-α -308 G > A genotype may be a potential genetic marker on HD patients with AVF thrombosis.