Mario H Hirata - Academia.edu (original) (raw)

Papers by Mario H Hirata

Research paper thumbnail of Rapid detection of 3500Q and 3531 mutations andMspI polymorphism in exon 26 at the apolipoprotein B gene

Journal of Clinical Laboratory Analysis, 2001

Several environmental and genetic factors are associated with high levels of cholesterol. Hyperch... more Several environmental and genetic factors are associated with high levels of cholesterol. Hypercholesterolemia is the main phenotype of Familial Defective Apolipoprotein B and Familial Hypercholesterolemia that are caused by mutations at the apolipoprotein (apo) B and LDL receptor genes, respectively. Identification of the specific genetic alteration associated with hypercholesterolemia is an important issue in clinical diagnosis of high risk for CAD. Apo B gene mutations and polymorphisms are usually screened by SSCP, DGGE, and heteroduplex, which must be confirmed by DNA sequencing or by direct detection using PCR techniques. In this study, we have optimized a PCR-RFLP procedure for identification of 3500Q and 3531 mutations and MspI polymorphism at the apo B gene. The technique can be performed in a single reaction, using the restriction endonuclease MspI for simultaneous detection of 3500Q mutation and MspI polymorphism, and NsiI for detection of 3531 mutation. The procedure was validated by analysis of control DNA samples from individuals carrying these mutations. Screening of 186 Brazilian hypercholesterolemic individuals showed that the frequency of the M-allele (7.8%) of MspI polymorphism was similar to that found in other individuals with CAD. However, neither 3500Q nor 3531 mutations were detected in this group. In conclusion, this procedure is simple and rapid, being easily introduced in clinical laboratories for direct detection of the more frequent mutations at the apo B gene associated with hypercholesterolemia.

Research paper thumbnail of Angiotensin I-converting enzyme gene polymorphism and drug response in familial hypercholesterolemia

Revista Brasileira De Ciencias Farmaceuticas, 2001

Research paper thumbnail of Influence of antidepressant drugs on DNA methylation of ion channels genes in blood cells of psychiatric patients

Epigenomics, Jul 1, 2022

Aim: This study investigated the influence of antidepressant drugs on methylation status of KCNE1... more Aim: This study investigated the influence of antidepressant drugs on methylation status of KCNE1, KCNH2 and SCN5A promoters and ECG parameters in adult psychiatric patients. Materials & methods: Electrocardiographic evaluation (24 h) and blood samples were obtained from 34 psychiatric patients before and after 30 days of antidepressant therapy. Methylation of promoter CpG sites of KCNE1, KCNH2 and SCN5A was analyzed by pyrosequencing. Results: Three CpG and four CpG sites of KCNE1 and SCN5A, respectively, had increased % methylation after treatment. Principal component analysis showed correlations of the methylation status with electrocardiographic variables, antidepressant doses and patient age. Conclusion: Short-term treatment with antidepressant drugs increase DNA methylation in KCNE1 and SCN5A promoters, which may induce ECG alterations in psychiatric patients.

Research paper thumbnail of Resposta terapêutica a fluvastatina é modulada pelo polimorfismo I/D do gene da enzima conversora da angiotensina-I em indivíduos com hipercolesterolemia primária

Revista Brasileira De Ciencias Farmaceuticas, 2001

Research paper thumbnail of Genetic characterization of patients with familial hypercholesterolemia from Brazil: a broad spectrum of mutations in the LDLR gene

Revista Brasileira De Ciencias Farmaceuticas, 2002

Research paper thumbnail of Effect of Pvu II polymorphism at the LDL receptor gene on response to fluvastatin treatment in patients with primary hypercholesterolemia

Research paper thumbnail of Estatinas modifican la expresión de genes del metabolismo intracelular del colesterol mediante la modulación de mecanismos epigenéticos

Revista chilena de cardiología, 2014

Research paper thumbnail of Are the polymorphisms in the methylenetetrahydrofolate reductase gene a risk factor for neural tube defects?: A preliminary report

Research paper thumbnail of Tecnicas de amplificacao de gene para o diagnostico da tuberculose

Revista de ciencias farmaceuticas, 1997

Resumo: O advento das técnicas de biologia molecular no diagnóstico laboratorial vem mudando o pa... more Resumo: O advento das técnicas de biologia molecular no diagnóstico laboratorial vem mudando o panorama de procedimentos clínico-terapêuticos. Discorreu-se, nesse artigo, sobre as várias formas de contribuiçöes da biologia molecular e particularmente de PCR ...

Research paper thumbnail of Influence of the low-density lipoprotein receptor gene on serun lipid levels response to fluvastatin treatment

Research paper thumbnail of Aplicacao de tecnicas de biologia molecular na avaliacao da resistencia de M.tuberculosis a drogas antimicrobianas

Revista de ciencias farmaceuticas, 1997

Research paper thumbnail of Novas mutações nos genes MYH7, MYBPC3 e TNNT2 relacionados à cardiomiopatia hipertrófica

Arquivos Brasileiros De Cardiologia, 2016

Research paper thumbnail of Differential regulation of apica/basolateral surface expression drug transporters by atorvastatin in caco-2 cells

Research paper thumbnail of Polymorphisms of low density lipoprotein receptor gene in individuals with heterozygous familial hypercholesterolemia from Brazil

Research paper thumbnail of Polymorphisms of the low-density lipoprotein receptor gene in Brazilian individuals with heterozygous familial hypercholesterolemia

Brazilian Journal of Medical and Biological Research, Nov 1, 2000

Research paper thumbnail of Impact of cholesterol on ABC and SLC transporters expression and function and its role in disposition variability to lipid-lowering drugs

Pharmacogenomics, Jun 1, 2009

This report focuses on the effects of cholesterol on the expression and function of the ATP-bindi... more This report focuses on the effects of cholesterol on the expression and function of the ATP-binding cassette (ABCB1, ABCG2 and ABCC2) and solute-linked carrier (SLCO1B1 and SLCO2B1) drug transporters with a particular focus on the potential impact of cholesterol on lipid-lowering drug disposition. Statins are the most active agents in the treatment of hypercholesterolemia. However, considerable interindividual variation exists in the response to statin therapy. Therefore, it would be huge progress if factors were identified that reliably differentiate between responders and nonresponders. Many studies have suggested that plasma lipid concentrations can affect drug disposition of compounds, such as ciclosporin and amphotericin B. Both compounds are able to affect the expression and function of ABC transporters. Although still speculative, these effects might be owing to the regulation of drug transporters by plasma cholesterol levels. Studies with normo- and hyper-cholesterolemic individuals, before and after atorvastatin treatment, have demonstrated that plasma cholesterol levels are correlated with drug transporter expression, as well as being related to atorvastatin’s cholesterol-lowering effect. The mechanism influencing the correlation between cholesterol levels and the expression and function of drug transporters remains unclear. Some studies provide strong evidence that nuclear receptors, such as the pregnane X receptor and the constitutive androstane receptor, mediate this effect. In the near future, pharmacogenomic studies with individuals in a pathological state should be performed in order to identify whether high plasma cholesterol levels might be a factor contributing to interindividual oral drug bioavailability.

Research paper thumbnail of Polimorfismo do gene da apolipoproteína e influência a resposta terapêutica a estatinas em indivíduos com hipercolesterolemia familial

Research paper thumbnail of Involvement of TLR2 and TLR4, <i>Chlamydophila pneumoniae</i> and <i>Mycoplasma pneumoniae</i> in adventitial inflammation of aortic atherosclerotic aneurysm

World Journal of Cardiovascular Diseases, 2014

Aortic atherosclerotic aneurysm (AAA) is associated with adventitial inflammation where infection... more Aortic atherosclerotic aneurysm (AAA) is associated with adventitial inflammation where infection is suggested to have a role. Co-infection with Chlamydophila pneumoniae (Cp) and Mycoplasma pneumoniae (Mp) was linked with coronary plaque rupture, in association with vessel dilatation and adventitial inflammation. Pathogens are recognized by Toll-like receptors (TLRs) development of the inflammatory process. Objective: Here, we studied whether co-infection by Cp and Mp was involved in the increased inflammation present in AAA and if it could be associated with deficient expression of TLRs. We compared human samples of AAA with non-dilated human aortic atherosclerotic lesions, regarding the amount of Cp and Mp antigens, and expression of TLR2 and TLR4. Methods: Two groups of aorta fragments were analyzed: G1 (n = 13) moderate atherosclerosis and G2 (n = 14) AAA samples, through immunohistochemistry and in situ hybridization methods. Results: Mp and Cp antigens in intima/medial layer were greater in G2 than G1, with no difference in adventitia. TLR2 and TLR4 were higher in G2 than G1 adventitia fat. There was a correlation between Mp versus TLR2 and of TLR4 in intima/medial layer and in adventitia of G1, but there was a lack of correlation in G2. In Cp adventitia, the correlation in G1 was high with TLR2 but not with TLR4, and in G2 the correlation was positive for both TLRs. Conclusion: This study favors the concept that symbiotic co-infection by Cp and Mp participates in the pathogenesis of AAA. It also emphasizes that adventitial fat is the initial site for colonization of these bacteria that probably reach the tissue through vasa vasorum. An exacerbated immune reaction is not efficient to control the infection that reaches and proliferates in high levels at the medial and intimal layer, contributing to the development of vessel dilatation.

Research paper thumbnail of Effect of sevoflurane on the inflammatory response during cardiopulmonary bypass in cardiac surgery: the study protocol for a randomized controlled trial

Trials, Jan 6, 2021

Background: Recent experimental evidence shows that sevoflurane can reduce the inflammatory respo... more Background: Recent experimental evidence shows that sevoflurane can reduce the inflammatory response during cardiac surgery with cardiopulmonary bypass. However, this observation so far has not been assessed in an adequately powered randomized controlled trial. Methods: We plan to include one hundred patients undergoing elective coronary artery bypass graft with cardiopulmonary bypass who will be randomized to receive either volatile anesthetics during cardiopulmonary bypass or total intravenous anesthesia. The primary endpoint of the study is to assess the inflammatory response during cardiopulmonary bypass by measuring PMN-elastase serum levels. Secondary endpoints include serum levels of other pro-inflammatory markers (IL-1β, IL-6, IL-8, TNFα), anti-inflammatory cytokines (TGFβ and IL-10), and microRNA expression in peripheral blood to achieve possible epigenetic mechanisms in this process. In addition clinical endpoints such as presence of major complications in the postoperative period and length of hospital and intensive care unit stay will be assessed. Discussion: The trial may determine whether adding volatile anesthetic during cardiopulmonary bypass will attenuate the inflammatory response. Trial registration: ClinicalTrials.gov NCT02672345. Registered on February 2016 and updated on June 2020.

Research paper thumbnail of Padronização da PCR em tempo real com curva de dissociação de alta resolução para detecção de mutações no gene rpoB relacionadas com resistência a rifampicina em isolados clínicos de M. tuberculosis

Research paper thumbnail of Rapid detection of 3500Q and 3531 mutations andMspI polymorphism in exon 26 at the apolipoprotein B gene

Journal of Clinical Laboratory Analysis, 2001

Several environmental and genetic factors are associated with high levels of cholesterol. Hyperch... more Several environmental and genetic factors are associated with high levels of cholesterol. Hypercholesterolemia is the main phenotype of Familial Defective Apolipoprotein B and Familial Hypercholesterolemia that are caused by mutations at the apolipoprotein (apo) B and LDL receptor genes, respectively. Identification of the specific genetic alteration associated with hypercholesterolemia is an important issue in clinical diagnosis of high risk for CAD. Apo B gene mutations and polymorphisms are usually screened by SSCP, DGGE, and heteroduplex, which must be confirmed by DNA sequencing or by direct detection using PCR techniques. In this study, we have optimized a PCR-RFLP procedure for identification of 3500Q and 3531 mutations and MspI polymorphism at the apo B gene. The technique can be performed in a single reaction, using the restriction endonuclease MspI for simultaneous detection of 3500Q mutation and MspI polymorphism, and NsiI for detection of 3531 mutation. The procedure was validated by analysis of control DNA samples from individuals carrying these mutations. Screening of 186 Brazilian hypercholesterolemic individuals showed that the frequency of the M-allele (7.8%) of MspI polymorphism was similar to that found in other individuals with CAD. However, neither 3500Q nor 3531 mutations were detected in this group. In conclusion, this procedure is simple and rapid, being easily introduced in clinical laboratories for direct detection of the more frequent mutations at the apo B gene associated with hypercholesterolemia.

Research paper thumbnail of Angiotensin I-converting enzyme gene polymorphism and drug response in familial hypercholesterolemia

Revista Brasileira De Ciencias Farmaceuticas, 2001

Research paper thumbnail of Influence of antidepressant drugs on DNA methylation of ion channels genes in blood cells of psychiatric patients

Epigenomics, Jul 1, 2022

Aim: This study investigated the influence of antidepressant drugs on methylation status of KCNE1... more Aim: This study investigated the influence of antidepressant drugs on methylation status of KCNE1, KCNH2 and SCN5A promoters and ECG parameters in adult psychiatric patients. Materials &amp; methods: Electrocardiographic evaluation (24 h) and blood samples were obtained from 34 psychiatric patients before and after 30 days of antidepressant therapy. Methylation of promoter CpG sites of KCNE1, KCNH2 and SCN5A was analyzed by pyrosequencing. Results: Three CpG and four CpG sites of KCNE1 and SCN5A, respectively, had increased % methylation after treatment. Principal component analysis showed correlations of the methylation status with electrocardiographic variables, antidepressant doses and patient age. Conclusion: Short-term treatment with antidepressant drugs increase DNA methylation in KCNE1 and SCN5A promoters, which may induce ECG alterations in psychiatric patients.

Research paper thumbnail of Resposta terapêutica a fluvastatina é modulada pelo polimorfismo I/D do gene da enzima conversora da angiotensina-I em indivíduos com hipercolesterolemia primária

Revista Brasileira De Ciencias Farmaceuticas, 2001

Research paper thumbnail of Genetic characterization of patients with familial hypercholesterolemia from Brazil: a broad spectrum of mutations in the LDLR gene

Revista Brasileira De Ciencias Farmaceuticas, 2002

Research paper thumbnail of Effect of Pvu II polymorphism at the LDL receptor gene on response to fluvastatin treatment in patients with primary hypercholesterolemia

Research paper thumbnail of Estatinas modifican la expresión de genes del metabolismo intracelular del colesterol mediante la modulación de mecanismos epigenéticos

Revista chilena de cardiología, 2014

Research paper thumbnail of Are the polymorphisms in the methylenetetrahydrofolate reductase gene a risk factor for neural tube defects?: A preliminary report

Research paper thumbnail of Tecnicas de amplificacao de gene para o diagnostico da tuberculose

Revista de ciencias farmaceuticas, 1997

Resumo: O advento das técnicas de biologia molecular no diagnóstico laboratorial vem mudando o pa... more Resumo: O advento das técnicas de biologia molecular no diagnóstico laboratorial vem mudando o panorama de procedimentos clínico-terapêuticos. Discorreu-se, nesse artigo, sobre as várias formas de contribuiçöes da biologia molecular e particularmente de PCR ...

Research paper thumbnail of Influence of the low-density lipoprotein receptor gene on serun lipid levels response to fluvastatin treatment

Research paper thumbnail of Aplicacao de tecnicas de biologia molecular na avaliacao da resistencia de M.tuberculosis a drogas antimicrobianas

Revista de ciencias farmaceuticas, 1997

Research paper thumbnail of Novas mutações nos genes MYH7, MYBPC3 e TNNT2 relacionados à cardiomiopatia hipertrófica

Arquivos Brasileiros De Cardiologia, 2016

Research paper thumbnail of Differential regulation of apica/basolateral surface expression drug transporters by atorvastatin in caco-2 cells

Research paper thumbnail of Polymorphisms of low density lipoprotein receptor gene in individuals with heterozygous familial hypercholesterolemia from Brazil

Research paper thumbnail of Polymorphisms of the low-density lipoprotein receptor gene in Brazilian individuals with heterozygous familial hypercholesterolemia

Brazilian Journal of Medical and Biological Research, Nov 1, 2000

Research paper thumbnail of Impact of cholesterol on ABC and SLC transporters expression and function and its role in disposition variability to lipid-lowering drugs

Pharmacogenomics, Jun 1, 2009

This report focuses on the effects of cholesterol on the expression and function of the ATP-bindi... more This report focuses on the effects of cholesterol on the expression and function of the ATP-binding cassette (ABCB1, ABCG2 and ABCC2) and solute-linked carrier (SLCO1B1 and SLCO2B1) drug transporters with a particular focus on the potential impact of cholesterol on lipid-lowering drug disposition. Statins are the most active agents in the treatment of hypercholesterolemia. However, considerable interindividual variation exists in the response to statin therapy. Therefore, it would be huge progress if factors were identified that reliably differentiate between responders and nonresponders. Many studies have suggested that plasma lipid concentrations can affect drug disposition of compounds, such as ciclosporin and amphotericin B. Both compounds are able to affect the expression and function of ABC transporters. Although still speculative, these effects might be owing to the regulation of drug transporters by plasma cholesterol levels. Studies with normo- and hyper-cholesterolemic individuals, before and after atorvastatin treatment, have demonstrated that plasma cholesterol levels are correlated with drug transporter expression, as well as being related to atorvastatin’s cholesterol-lowering effect. The mechanism influencing the correlation between cholesterol levels and the expression and function of drug transporters remains unclear. Some studies provide strong evidence that nuclear receptors, such as the pregnane X receptor and the constitutive androstane receptor, mediate this effect. In the near future, pharmacogenomic studies with individuals in a pathological state should be performed in order to identify whether high plasma cholesterol levels might be a factor contributing to interindividual oral drug bioavailability.

Research paper thumbnail of Polimorfismo do gene da apolipoproteína e influência a resposta terapêutica a estatinas em indivíduos com hipercolesterolemia familial

Research paper thumbnail of Involvement of TLR2 and TLR4, <i>Chlamydophila pneumoniae</i> and <i>Mycoplasma pneumoniae</i> in adventitial inflammation of aortic atherosclerotic aneurysm

World Journal of Cardiovascular Diseases, 2014

Aortic atherosclerotic aneurysm (AAA) is associated with adventitial inflammation where infection... more Aortic atherosclerotic aneurysm (AAA) is associated with adventitial inflammation where infection is suggested to have a role. Co-infection with Chlamydophila pneumoniae (Cp) and Mycoplasma pneumoniae (Mp) was linked with coronary plaque rupture, in association with vessel dilatation and adventitial inflammation. Pathogens are recognized by Toll-like receptors (TLRs) development of the inflammatory process. Objective: Here, we studied whether co-infection by Cp and Mp was involved in the increased inflammation present in AAA and if it could be associated with deficient expression of TLRs. We compared human samples of AAA with non-dilated human aortic atherosclerotic lesions, regarding the amount of Cp and Mp antigens, and expression of TLR2 and TLR4. Methods: Two groups of aorta fragments were analyzed: G1 (n = 13) moderate atherosclerosis and G2 (n = 14) AAA samples, through immunohistochemistry and in situ hybridization methods. Results: Mp and Cp antigens in intima/medial layer were greater in G2 than G1, with no difference in adventitia. TLR2 and TLR4 were higher in G2 than G1 adventitia fat. There was a correlation between Mp versus TLR2 and of TLR4 in intima/medial layer and in adventitia of G1, but there was a lack of correlation in G2. In Cp adventitia, the correlation in G1 was high with TLR2 but not with TLR4, and in G2 the correlation was positive for both TLRs. Conclusion: This study favors the concept that symbiotic co-infection by Cp and Mp participates in the pathogenesis of AAA. It also emphasizes that adventitial fat is the initial site for colonization of these bacteria that probably reach the tissue through vasa vasorum. An exacerbated immune reaction is not efficient to control the infection that reaches and proliferates in high levels at the medial and intimal layer, contributing to the development of vessel dilatation.

Research paper thumbnail of Effect of sevoflurane on the inflammatory response during cardiopulmonary bypass in cardiac surgery: the study protocol for a randomized controlled trial

Trials, Jan 6, 2021

Background: Recent experimental evidence shows that sevoflurane can reduce the inflammatory respo... more Background: Recent experimental evidence shows that sevoflurane can reduce the inflammatory response during cardiac surgery with cardiopulmonary bypass. However, this observation so far has not been assessed in an adequately powered randomized controlled trial. Methods: We plan to include one hundred patients undergoing elective coronary artery bypass graft with cardiopulmonary bypass who will be randomized to receive either volatile anesthetics during cardiopulmonary bypass or total intravenous anesthesia. The primary endpoint of the study is to assess the inflammatory response during cardiopulmonary bypass by measuring PMN-elastase serum levels. Secondary endpoints include serum levels of other pro-inflammatory markers (IL-1β, IL-6, IL-8, TNFα), anti-inflammatory cytokines (TGFβ and IL-10), and microRNA expression in peripheral blood to achieve possible epigenetic mechanisms in this process. In addition clinical endpoints such as presence of major complications in the postoperative period and length of hospital and intensive care unit stay will be assessed. Discussion: The trial may determine whether adding volatile anesthetic during cardiopulmonary bypass will attenuate the inflammatory response. Trial registration: ClinicalTrials.gov NCT02672345. Registered on February 2016 and updated on June 2020.

Research paper thumbnail of Padronização da PCR em tempo real com curva de dissociação de alta resolução para detecção de mutações no gene rpoB relacionadas com resistência a rifampicina em isolados clínicos de M. tuberculosis