ABSTRACT
OBJECTIVE@#To explore the correlation of cytochrome B-245 alpha chain (CYBA) rs4673 and cholesteryl ester transfer protein (CETP) rs12720922 polymorphisms with the susceptibility of gene-ralized aggressive periodontitis (GAgP).@*METHODS@#The study was a case-control trial. A total of 372 GAgP patients and 133 periodontally healthy controls were recruited. The CYBA rs4673 and CETP rs12720922 polymorphisms were detected by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Logistic regression models were used to analyze the correlation of CYBA rs4673 and CETP rs12720922 variants with the susceptibility of GAgP. The interaction between the two gene polymorphisms to the susceptibility of GAgP was analyzed by the likelihood ratio test. The interaction model adopted was the multiplication model.@*RESULTS@#The mean age of GAgP group and control group was (27.5±5.2) years and (28.8±7.1) years respectively. There was significant difference in age between the two groups (P < 0.05). The gender distribution (male/female) was 152/220 and 53/80 respectively, and there was no significant difference between GAgP group and controls (P>0.05). For CYBA rs4673, the frequency of CT/TT genotype in the GAgP group was significantly higher than that in the controls [18.0% (66/366) vs. 10.6% (14/132), P < 0.05]. After adjusting age and gender, the individuals with CT/TT genotype had a higher risk of GAgP (OR=1.86, 95%CI: 1.01-3.45, P < 0.05), compared with CC genotype. There was no statistically significant difference in distributions of the CETP rs12720922 genotypes (GG, AA/AG) between GAgP patients and healthy controls (P>0.05). A significant interaction between CYBA rs4673 and CETP rs12720922 in the susceptibility to GAgP was observed. The GAgP risk of the individuals with CYBA rs4673 CT/TT and CETP rs12720922 GG genotypes was significantly increased (OR=3.25, 95%CI: 1.36-7.75, P < 0.01), compared with those carrying CC and AA/AG genotypes.@*CONCLUSION@#CYBA rs4673 CT/TT genotype is associated with GAgP susceptibility. There is a significant interaction between CYBA rs4673 CT/TT genotype and CETP rs12720922 GG genotype in the susceptibility of GAgP.
Subject(s)
Adult , Female , Humans , Male , Young Adult , Aggressive Periodontitis/genetics , Case-Control Studies , Cholesterol Ester Transfer Proteins/genetics , Cytochrome b Group , Gene Frequency , Genetic Predisposition to Disease , Genotype , NADPH Oxidases/genetics , Polymorphism, Single NucleotideABSTRACT
BACKGROUND: Most cases with congenital hypothyroidism (CH) are usually sporadic, while about 20% of the cases are caused by genetic defects. Little information is available regarding the mutation incidence and genetic heterogeneity of CH in Koreans. We aimed to determine the mutation incidence of CH in newborn screenings (NBS) and to evaluate the frequency and spectrum of mutations underlying CH. METHODS: A total of 112 newborns with thyroid dysfunction were enrolled from 256,624 consecutive NBS. Furthermore, 58 outpatients with primary CH were added from an endocrine clinic. All coding exons of TSHR, PAX8, TPO, DUOX2, DUOXA2, and SCL5A5 were sequenced. RESULTS: The mutation incidence of CH was estimated to be 1 in 6,580 newborns. A total of 36 different mutations were identified in 53 cases. The overall mutation positive rate was 31%. The DUOX2 mutations were the most prevalent in both newborns and outpatients. Seven different recurrent mutations [p.G488R (n=13), p.A649E (n=3), p.R885Q (n=3), p.I1080T (n=2), and p.A1206T (n=2) in DUOX2; p.Y138X (n=9) in DUOXA2; and p.R450H (n=5) in TSHR) were identified as the mutations underlying CH. CONCLUSIONS: The mutation incidence of CH was considerably higher than expected in the Korean newborn population. This study revealed seven different recurrent mutations underlying CH. We conclude that DUOX2 mutations are a frequent cause of CH in the Korean population.
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Asian People/genetics , Congenital Hypothyroidism/epidemiology , Exons , Genetic Association Studies , Genotype , NADPH Oxidases/genetics , Polymorphism, Single Nucleotide , Republic of Korea/epidemiology , Sequence Analysis, DNA , Thyrotropin/bloodABSTRACT
Professor Alessandri died in 1980. We started our residency in Internal Medicine about 30 years later. Considering the profound changes our society has witnessed, including medical practice, I would like to approach the meaning of his work for our generation. It is not the Father’s figure nor his Aura what inspires us today. Neither is his personality nor his shape. His universality comes from his transcendent image as a teacher. Today’s teachers live rough times, their social status has changed, their professional requirements have grown exponentially, they have to adapt to social phenomena like the Internet and multiculturalism. Being a teacher nowadays demands to be a multifaceted expert. Things have changed since Professor Alessandri made rounds with his patients. But a deeper look allows us to understand that everything returns to where it started: professional deontology of the teacher, never fading but transcendent. We know that Doctor Alessandri had the natural gift to keep faithful to that code with consistency and perseverance. He excelled with integrity in every aspect including professional betterment, constructive work for his institution, collegiality, a warm relationship with students and a model of social values. Beyond virtues and personal defects he will keep on being the mould in which present teachers should be formed, engraved in their souls and in the subconscious of students that seek to learn.
Subject(s)
Animals , Dogs , Humans , Mice , /metabolism , Oxygen/chemistry , Sick Sinus Syndrome/genetics , Sinoatrial Node/pathology , Angiotensin II/metabolism , Apoptosis , Biomarkers/metabolism , Electrocardiography/methods , Mice, Transgenic , NADPH Oxidases/genetics , Reactive Oxygen Species , Sick Sinus Syndrome/metabolismABSTRACT
Chronic granulomatous disease is a primary immunodeficiency caused by mutations in the genes encoding subunits of the phagocytic NADPH oxidase system. Patients can present with severe, recurrent infections and noninfectious conditions. Among the latter, inflammatory manifestations are predominant, especially granulomas and colitis. In this article, we systematically review the possible mechanisms of hyperinflammation in this rare primary immunodeficiency condition and their correlations with clinical aspects.
Subject(s)
Humans , Granulomatous Disease, Chronic , NADPH Oxidases/genetics , Neutrophils/immunology , Granulomatous Disease, Chronic/genetics , Granulomatous Disease, Chronic/immunology , Granulomatous Disease, Chronic/microbiology , Inflammation Mediators/physiology , NADPH Oxidases/deficiency , Neutrophils/microbiology , Reactive Oxygen Species/immunology , Reactive Oxygen Species/metabolismABSTRACT
Introducción: La Enfermedad Granulomatosa Crónica (EGC) se presenta como consecuencia de mutaciones en los genes que codifican 5 de las subunidades del sistema NADPH oxidasa humano. Su forma más común es causada por cambios en el gen CYBB que codifica gp91 phox. Objetivo: Identificar el defecto molecular que lleva a la presentación de la EGC. Caso clínico: Paciente de sexo masculino con antecedentes de enfermedad diarreica aguda y abscesos perianales recurrentes desde los 2 meses. A los 6 meses, presentó una inflamación crónica del colon y colitis bacteriana. A los 3 años tenía infecciones en las vías respiratorias inferiores y perianales. Estudio compatible con EGC. El análisis del ADNc identificó expresión anormal del ARNm, lo cual se confirmó al realizar la secuenciación. Específicamente se observó la ausencia del exón 2. Adicionalmente, los datos de la secuenciación del ADNg identificaron una alteración en el sitio aceptor de "splicing" del intrón 1, que incluye una deleción seguida de la inserción de 3 nucleótidos (c.46-14_-11delTTCT insGAA). Conclusiones: Se presenta el primer estudio molecular de un paciente con EGC por defecto de "splicing" reportado en Colombia. La definición de la mutación y su correlación con el fenotipo es importante para proveer una apropiada consejería genética al paciente y su familia.
Chronic granulomatous disease (CGD) is caused by mutations in the genes that encode five of the subunits of the human NADPH oxidase. The most common form is caused by mutations in CYBB, the human gene encoding gp 91 phox. Objective: To identify the molecular defects causing CGD. Case report: A male patient with a history of acute diarrhea and recurrent perianal abscess since two months old. At 6 months, the patient presented a chronic inflammatory disease of the colon and bacterial colitis. After three years, he developed infections in the lower and perianal respiratory tract. The cDNA analysis identified abnormal mRNA expression, which was confirmed by sequencing. Specifically the exclusion of exon 2 was observed. Additionally, gDNA sequencing identified an alteration in the acceptor splice site of intron 1, including a deletion followed by insertion of three nucleotides (c.46-14_-11delTTCT insGAA). Conclusions: The first molecular study of a patient with CGD due to splicing pattern change, reported in Colombia, is presented. The definition of the mutation and its correlation with the phenotype is essential to provide appropriate genetic counseling to patients and their families.
Subject(s)
Humans , Male , Infant , Chromosomes, Human, X , Granulomatous Disease, Chronic/genetics , Mutation , NADPH Oxidases/genetics , DNA, Complementary/genetics , RNA, Messenger/genetics , Base Sequence , Cell Separation , Exons , Granulomatous Disease, Chronic/diagnosis , Flow Cytometry , Phenotype , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , RNA SplicingABSTRACT
Objective : This study aimed to determine whether a hypercholesterolemic diet induces hepatic steatosis, alterations in mRNA expression of NADPH oxidase subunits, and antioxidant defenses.Materials and methods : Fischer rats were divided into two groups of eight animals according to the treatment, control (C) and hypercholesterolemic diet (H). Those in group C were fed a standard diet (AIN-93M), and those of the group H were fed a hypercholesterolemic diet (25% soybean oil and 1% cholesterol).Results : The hypercholesterolemic diet did not affect body weight, but resulted in the accumulation of lipids in the liver, increased serum activities of aminotransferases and cholesterol levels. Biomarker of lipid peroxidation (TBARS) and mRNA expression of NADPH oxidase subunits p22phox and p47phox were increased in the liver of animals in group H. Besides, the activity and expression of antioxidant enzymes were altered.Conclusion : The results show increased mRNA expression of NADPH oxidase subunits and changes in antioxidant enzyme activities in diet-induced hepatic steatosis. Arq Bras Endocrinol Metab. 2014;58(3):251-9.
Objetivo Determinar se uma dieta hipercolesterolemiante induz esteatose hepática, alterações na expressão de mRNA da NADPH oxidase e nas defesas antioxidantes.Materiais e métodos : Ratas Fischer foram divididas em dois grupos de oito animais de acordo com o tratamento recebido, controle (C) e hipercolesterolêmico (H). Aquelas do grupo C foram alimentadas com dieta padrão (AIN-93M) e as do grupo H foram alimentadas com dieta hipercolesterolemiante (25% de óleo de soja e 1% de colesterol). As dietas foram oferecidas por oito semanas.Resultados : O grupo H apresentou acúmulo de lipídios no fígado, aumento das atividades de ALT e AST e da concentração de colesterol no soro comparado ao grupo C. O marcador da peroxidação lipídica (TBARS) e os níveis de mRNA das subunidades p47phox da NADPH-oxidase e p22phox foram aumentados no fígado de animais do grupo H, além de alteração da atividade e expressão de enzimas antioxidantes.Conclusão : Os resultados mostram um aumento na expressão de subunidades da NADPH oxidase e alterações na atividade das enzimas antioxidantes na esteatose hepática induzida por dieta hipercolesterolemiante. Arq Bras Endocrinol Metab. 2014;58(3):251-9.
Subject(s)
Animals , Female , Cholesterol, Dietary/adverse effects , Fatty Liver/etiology , Hypercholesterolemia/etiology , Liver/enzymology , NADPH Oxidases/genetics , RNA, Messenger/metabolism , Alanine Transaminase/blood , Antioxidants/analysis , Aspartate Aminotransferases/blood , Body Weight , Catalase/metabolism , Cholesterol/blood , Diet, High-Fat/adverse effects , Glutathione/analysis , Lipids/blood , NADPH Oxidases/metabolism , Oxidative Stress , Real-Time Polymerase Chain Reaction , Superoxide Dismutase/metabolism , Triglycerides/bloodABSTRACT
La actividad de Rho kinasa (ROCK) cardíaca en la hipertensión arterial (HTA) y el efecto del tratamiento antihipertensivo conjunto han sido poco estudiados. Hemos planteado que la adición de un inhibidor de ROCK al tratamiento antihipertensivo convencional podría tener efectos preventivos adicionales al uso aislado del antihipertensivo. Objetivo: Determinar la actividad de ROCK ventricular y parámetros de remodelamiento cardíaco en ratas hipertensas con y sin tratamiento antihipertensivo, adicionando un inhibidor directo de ROCK. Métodos. Se usaron ratas Sprague Dawley de 150 grs. ( n = 12 - 13/grupo) unifrectomizadas tratadas con desoxicorticosterona (DOCA, 100 mg/Kg/sem sbc) durante 6 semanas. Como controles se usaron ratas unifrectomizadas. Otros 3 grupos recibieron DOCA y además el antagonista del receptor de angiotensina n, candesartán (10 mg/kg/día) o el inhibidor de la vía ROCK fasudil (50 mg/Kg/dia), o la combinación de ambos (5 y 25 mg/Kg/dia, respectivamente), vía gavage desde la tercera semana post cirugía, durante 3 semanas. Al finalizar los tratamientos se determinó la masa corporal (MC), presión arterial sistólica (PAS) y la masa cardíaca relativa (MCR). Además se midió en el ventrículo izquierdo la fosforilación de la fosfatasa de la miosina (MYPT-1) como índice de activación de ROCK, la infiltración de macrófagos/ monocitos (células ED1 positivas), la expresión proteica de colágeno I (por Western blot) y la expresión génica de la subunidad gp91 de NADPH oxidasa y eNOS por RTPCR. Resultados: Con respecto de las ratas sham, en las ratas hipertensas se observó hipertrofia cardiaca de 63 por ciento (p < 0,05), junto a aumento significativo (p < 0,05) de MYPT-1 fosforilado/total (300 por ciento), de colágeno miocárdico I (en 14 veces), de células ED1 en 270 por ciento y de la expresión génica de la subunidad gp91 de NADPH...
Background: The effect of cardiac Rho-kinase (ROCK) on hypertension (HT) and cardiac hypertrophy prevention and also the combined anti-hypertensive treatment have been scarcely studied. We hypothesized that the addition of a ROCK inhibitor to conventional anti-hypertensive treatment may have additional beneficial effects. Ainv to determine ventricular ROCK activity and ventricular remodeling in hypertensive rats treated with Angiotensin II inhibition with the addition of a ROCK inhibitor. Methods: Sprague-Dawley rats weighing 150 grams had one kidney removed and received deoxycortisterone acétate (DOCA, 100 mg/kg/week, during 6 weeks). Unilaterally nephrectomized rats were used as controls. The other 3 groups received DOCA along with the Angiotensin II receptor blocker candesartan (10 mg/kg/day) or the combination of both agents (5 and 25 mg/kg/day, respectively) and ROCK inhibitor fasudil (50 mg/kg/day) for 3 weeks starting 3 weeks after surgery. Body mass (BM), systolic blood pressure (SBP) and relative cardiac mass (RCM) were measured. In addition, myosin phosphatase (MYPT-1) phosphorylation was measured as an indicator of ROCK activation. Cardiac infiltration of macrophages/monocytes (ED1 positive cells), collagen I protein contení (by Western Blot) and also cardiac gene expression of NADPH oxydase GP91 subunit and eNOS were determined by RT-PCR. Results: In hypertensive rats we observed cardiac hypertrophy by 63 percent (p < 0.05), a 300 percent increase in cardiac MYPT-1 phosphorylation (p< 0.05), 14 times increase in myocardial collagen type 1,270 percent increase in ED1 cells, a 75 percent increased gene expression of NADPH oxydase GP91 subunit and a 37 percent reduction (p< 0.05) in the gene expression of cardiac eNOS. In hypertensive DOCA rats treated during 3 week with candesartan, fasudil or the combination of both, we observed a significant reduction in cardiac hypertrophy and normalization of SBP, MYPT-1 phosphorylation, ...
Subject(s)
Animals , Rats , /administration & dosage , Benzimidazoles/administration & dosage , Angiotensin II Type 1 Receptor Blockers/administration & dosage , Hypertrophy, Left Ventricular/prevention & control , rho-Associated Kinases/antagonists & inhibitors , Tetrazoles/administration & dosage , /analogs & derivatives , Blotting, Western , Drug Therapy, Combination , Gene Expression , Hypertension/drug therapy , NADPH Oxidases/genetics , Polymerase Chain Reaction , Rats, Sprague-DawleyABSTRACT
Uno de los objetivos del tratamiento antihipertensivo, más allá de normalizar las cifras tensionales, es disminuir/regresar el remodelado patológico hipertensivo cardiovascular y renal, de manera de reducir eficientemente el riesgo dado por esta patología. Al respecto, la vía de señalización intracelular de la Rho kinasa (ROCK) es un mecanismo de vasoconstricción y de promoción de remodelado que podría ser un blanco atractivo en el tratamiento antihipertensivo. Objetivo: Evaluar a nivel vascular los niveles de TGF beta, la expresión génica vascular de NADPH oxidasa (fuente de stress oxidativo vascular), y el grado de inflamación parietal y su dependencia de ROCK en la hipertensión arterial (HTA) experimental. Métodos: Se compararon 5 grupos experimentales. Se usó el modelo de HTA por administración de deoxicorticosterona (DOCA, 100mg/Kg, sc una vez/semana) + sal en ratas Sprague Dawley uninefrectomizadas. Como controles, se usaron ratas uninefrectomizadas (Sham). Las ratas DOCA se randomizaron para recibir el inhibidor específico de ROCK fasudil (Fas, 50 mg/kg/d, por gavage, durante 3 semanas), desde la semana 3 post cirugía, o candesartán (Cand, 10 mg/kg/d, por gavage, durante 3 semanas), o fasudil (25 mg/kg/d) + candesartan (5 mg/ kg/d por gavage, 3 semanas ). Al finalizar los experimentos se midieron en la aorta los niveles de MYPT1 fosforilada/total, blanco de ROCK y estimador de su activación (por Western blot), de TGF beta (por Western blot), los niveles de mRNA de las subunidades p22 Phox y gp 91 de la NADPH oxidasa (por RT PCR) y el número de células infamatorias ED1 en anillos aórticos (por inmunohistoquímica). Resultados: La presión arterial sistólica aumentó en las ratas DOCA a 172 +/- 7 mm Hg (p < 0,05) y fue normalizada después de 3 semanas de tratamiento con candesartán, fasudil y de candesartán + fasudil. La actividad de ROCK en aorta aumentó en 4 veces en las ratas hipertensas (p < 0,05)...
Background: Rho kinase (ROCK) activity promotes vasoconstriction and pathological vascular remodeling in experimental hypertension. Our working hypothesis is that ROCK inhibition could be an attractive target to prevent vascular remodeling in hypertension. Objectives: We evaluated vascular TGF beta, the genic expression of NADPH oxydase (a vascular oxidative stress source) and its dependency from ROCK activation in experimental hypertension in the rat. Methods: Five experimental groups were compared. Hypertension was induced by the administration of salt and deoxycorticosterone acetate (DOCA, 100 mg/Kg, weekly) to unilaterally nephrectomized rats. Unilaterally nephrectomized rats were used as controls (controls). DOCA rats were randomized to receive either Fasudil (a ROCK inhibitor, 50 mg/Kg/day) or candesartan (CAND, 10 mg/Kg/day for 3 weeks), starting 3 weeks after surgery. The other group received fasudil (25 mg/Kg/day) plus CAND (5 mg/Kg/day) for 3 weeks. After treatment, phosphorilated MYPT1 (a ROCK target expressing ROCK activation) was measured in aortic wall rings by Western Blot. We also determined TGF-beta (Western Blot), p22 Phox and gp 91subnits of NADPH oxydase mRNA (RT-PCR) and the number of ED1 infammatory cells. Results: In DOCA rats, SBP increased to 172 +/- 7 mm Hg (p < 0,05), and returned to normal values after 3 weeks with candesartan, fasudil or both combined. In these rats, ROCK activity in aorta was increased 4 times (p < 0,05) and returned to control values in the 3 groups receiving treatment. p22 Phox and gp 91subnits of NADPH oxydase mRNA were increased by 80 and 90 percent, respectively (p<0,05). These changes were reduced to control values in rats receiving fasudil and candesartan + fasudil. Gene expression of TGF-beta increased 4 times, and the number of ED1...
Subject(s)
Animals , Rats , /administration & dosage , Antihypertensive Agents/administration & dosage , Benzimidazoles/administration & dosage , Hypertension/drug therapy , rho-Associated Kinases/antagonists & inhibitors , Tetrazoles/administration & dosage , /analogs & derivatives , Blotting, Western , Enzyme Inhibitors , Gene Expression , Protein Kinase Inhibitors , NADPH Oxidases/genetics , NADPH Oxidases/metabolism , Polymerase Chain Reaction , Protein Serine-Threonine Kinases/antagonists & inhibitors , Rats, Sprague-Dawley , RNA, Messenger , Transforming Growth Factor betaABSTRACT
Vários estudos destacam as espécies reativas de oxigênio e nitrogênio (ERONs) como importantes contribuintes na patogênese de numerosas doenças cardiovasculares, incluindo hipertensão, aterosclerose e falência cardíaca. Tais espécies são moléculas altamente bioativas e com vida curta derivadas, principalmente, da redução do oxigênio molecular. O complexo enzimático da NADPH oxidase é a maior fonte dessas espécies reativas na vasculatura. Sob condições fisiológicas, a formação e eliminação destas substâncias aparecem balanceadas na parede vascular. Durante o desbalanço redox, entretanto, há um aumento na atividade da NADPH oxidase e predomínio de agentes pró-oxidantes, superando a capacidade de defesa orgânica antioxidante. Além disso, tal hiperatividade enzimática reduz a biodisponibilidade do óxido nítrico, crucial para a vasodilatação e a manutenção da função vascular normal. Apesar de a NADPH oxidase relacionar-se diretamente à disfunção endotelial, foi primeiramente descrita por sua expressão em fagócitos, onde sua atividade determina a eficácia dos mecanismos de defesa orgânica contra patógenos. As sutis diferenças existentes entre as unidades estruturais das NADPH oxidases, a depender do tipo celular que as expressa, podem ter implicações terapêuticas, permitindo a inibição seletiva do desequilíbrio redox induzido pela NADPH oxidase, sem comprometer, entretanto, sua participação nas vias fisiológicas de sinalização celular que garantem a proteção contra microorganismos.
Several studies refer to reactive oxygen and nitrogen species (RONS) as important agents in the pathogenesis of a number of heart diseases, including high blood pressure, arteriosclerosis and heart failure. Such species are highly bioactive molecules and a short life due chiefly to reduction of molecular oxygen. The enzyme complex of NADPH oxidase is the main source of these reactive species in vascular system. Under physiological conditions, formation and elimination of these substances seem balanced in vascular wall. During redox Unbalance, nonetheless, there is increase in NADPH oxidase activity and predominance of pro-oxidizing agents, surpassing the anti-oxidant capacity of the organism self-defense. Besides this, such enzyme hyperactivity reduces the bioavailability of nitric oxide, capital for vasodilation and maintenance of normal vascular function. In spite of NADPH oxidase being directly connected to the endothelial dysfunction, it was firstly described as for its expression in phagocytes, where its activity determines efficiency of organism defense mechanisms against pathogens. Slight differences between structural units of NADPH oxidases, depending on the type of cell which expresses it, may create therapeutic implications, allowing to selectively inhibiting redox unbalance triggered by NADPH oxidase, without compromising, however, its participation in physiological cellular signaling which make sure protection against micro-organisms.
Subject(s)
Humans , Hypertension/therapy , NADPH Oxidases/antagonists & inhibitors , Blood Pressure/physiology , Hypertension/enzymology , NADPH Oxidases/genetics , Oxidation-ReductionABSTRACT
The action mode of 4,4'-diaminodiphenylsulfone (DDS) is still under debate, although it has long been used in treatment of several dermatologic diseases including Hansen's disease. In this study, we tested the effect of DDS as an antioxidant on paraquat-induced oxidative stress in non-phagocytic human diploid fibroblasts (HDFs). Overall, preincubation of HDFs with DDS prevented the oxidative stress and the resulting cytotoxic damages caused by paraquat in these cells. The specific effects of DDS in paraquat-treated HDFs are summarized as follows: a) reducing the expression of NADPH oxidase 4 (NOX4) by inhibiting paraquat-induced activation of PKC; b) inhibiting paraquat-induced decreases in mitochondrial complex protein levels as well as in membrane potentials; c) consequently, inhibiting the generation of cytosolic and mitochondrial superoxide anions. Taken together, these findings suggest that DDS would suppress the radical generation in non-phagocytic HDFs during oxidative stress, and that DDS might have the extended potential to be used further in prevention of other oxidative stress-related pathologies.
Subject(s)
Humans , Male , Biphenyl Compounds/metabolism , Cell Death/drug effects , Cell Survival/drug effects , Dapsone/pharmacology , Diploidy , Enzyme Activation/drug effects , Fibroblasts/cytology , Free Radical Scavengers/metabolism , Gene Expression Regulation, Enzymologic/drug effects , Mitochondria/drug effects , NADPH Oxidases/genetics , Paraquat/toxicity , Phagocytosis/drug effects , Picrates/metabolism , Protein Kinase C/metabolism , RNA, Messenger/genetics , Reactive Oxygen Species/metabolism , Superoxides/metabolismABSTRACT
O sistema renina-angiotensina e o estresse oxidativo têm participação importante na fisiopatologia das complicações crônicas do diabetes. No presente estudo, foram avaliados 103 pacientes com diabetes tipo 1 (DM1) com idade de 28,8+-10,6 anos e duração de doença de 13,1+-8,5 anos e 158 controles não diabéticos quanto à presença dos polimorfismos I/D da ECA e C242T do p22phox, componente essencial para a ativação da NADPH oxidase. Esta análise de restrição enzimática para avaliação do polimorfismo C242T p22phox. Ambas as distribuições genotípicas obedeciam ao princípio do equilíbrio de Hardy-Weinberg. Os pacientes diabéticos foram submetidos a avaliação clínica e laboratorial quanto à presença de fatores associados ao risco de complicações (história de tabagismo e antecedentes familiares de diabetes tipo 2, dose diária de insulina, níveis pressóricos, índice de massa corporal, relação cintura-quadril, excreção urinária de albumina, taxa de filtração glomerular, perfil lipídico, controle glicêmico, níveis de proteína C-reativa) e rastreados quanto à presença de nefropatia diabética, considerada presença de micro ou macroalbuminúria; retinopatia diabética não proliferativa ou proliferativa e hipertensão arterial. Não houve diferença significativa entre a presença dos alelos D e I da ECA ou C e T do p22phox entre diabéticos e controles. Os polimorfismos avaliados não apresentaram associação com a presença de nefropatia, retinopatia ou hipertensão arterial. Pacientes portadores do alelo D apresentaram maiores níveis de pressão arterial diastólica (72,2+-12,3 vs 65,4+-11,6 mmHg, p=0,047) e proteína C-reativa comparados aos portadores do genótipo II [0,18 (0,04-0,38) vs 0,09 (0,04-0,16) mg/dl, p=0,05], porém ambas as análises perderam significância estatística após correção para duração do diabetes. A combinação dos polimorfismos não esteve associada à presença de complicações microvasculares ou hipertensão arterial. Concluímos que, na população de diabéticos ...
The renin-angiotensin system and the oxidative stress play an important role in the pathogenesis of the diabetic complications. In the present study 103 patients with type 1 diabetes (T1DM) aged 28.8+-10.6 years and with a disease duration of 13.1+-8.5 years and 158 non-dabetic controls were evaluated for the presence of the I/D polymorphism of the angiotensin converting enzyme (ACE) and the C242T polymorphism of the p22phox, an essential component for NADPH oxidase activation. The analysis was performed using polymerase chain reaction for both polymorphisms, followed by enzymatic restriction for C242T p22phox polymorphism. Genotypic distributions of both polymorphisms were in Hardy-Weinberg equilibrium. Diabetic patients underwent clinical and laboratory evaluation for the presence of risk factors associated with complications of diabetes (smoking and family history of type 2 diabetes, daily insulin dose, blood pressure, body mass index, waist - hip ratio, urinary albumin excretion, glomerular filtration rate, lipid profile, glycemic control, C-reactive protein levels) and screened for the presence of diabetic nephropathy, considered as the presence of micro or macroalbuminuria, diabetic retinopathy and hypertension. There was no significant difference between the presence of ACE D or I allele and p22phox C or T allele between diabetic patients and controls. The evaluated polymorphisms were not associated with the presence of nephropathy, retinopathy of hypertension. Patients with the D allele showed higher levels of diastolic blood pressure (72.2+-12.3 vs 65.4+-11.6 mmHg, p=0.047) and C-reactive protein compared with those carrying the II genotype [0.18 (0.04-0.38) vs 0.09 (0.04-0.16) mg/dl, p=0.05], but both analysis lost statistical significance after correction for duration of diabetes. The combination of both polymorphisms was not associated with microvascular complications or hypertension. We conclude that in the studies population of type ...
Subject(s)
Humans , Male , Female , Diabetes Mellitus, Type 1/complications , Diabetes Mellitus, Type 1/physiopathology , Cardiovascular Diseases/epidemiology , Cardiovascular Diseases/genetics , NADPH Oxidases/genetics , Oxidative Stress , Polymorphism, Genetic , Peptidyl-Dipeptidase A/genetics , Renin-Angiotensin System , Polymerase Chain Reaction , Risk FactorsABSTRACT
Chronic granulomatous disease (CGD) is a rare hereditary disorder characterized by recurrent life-threatening bacterial and fungal infections. The underlying defect in CGD is an inability of phagocytes to produce reactive oxygen species as a result of defects in NADPH oxidase. Considering that CGD generally affects about 3-4 in 1,000,000 individuals, it is surprising that the prevalence of CGD on Jeju Island is 20.7 in 1,000,000 individuals. We performed genetic analysis on 12 patients from 10 unrelated families and found that all patients had an identical homozygous single-base substitution of C to T in exon 1 (c.7C>T) of the CYBA gene, which was expected to result in a nonsense mutation (p.Q3X). Because Jeju Island has long been a geologically isolated region, the high prevalence of CGD on Jeju Island is presumably associated with an identical mutation inherited from a common ancestor or proband.
Subject(s)
Adolescent , Animals , Child , Child, Preschool , Female , Humans , Infant , Male , Young Adult , Asian People/genetics , Base Sequence , DNA Mutational Analysis , Geography , Granulomatous Disease, Chronic/genetics , Korea , Molecular Sequence Data , Mutation , NADPH Oxidases/genetics , PedigreeSubject(s)
Cell Separation , DNA Mutational Analysis , Flow Cytometry/methods , Granulomatous Disease, Chronic/diagnosis , Humans , Immunologic Tests/methods , Infant , Male , Membrane Glycoproteins/genetics , Mutation , NADPH Oxidases/genetics , Neutrophils/immunology , Pedigree , Rhodamines , Sensitivity and Specificity , X Chromosome Inactivation/immunologyABSTRACT
X-linked chronic granulomatous disease (X-CGD) is an immunodeficiency disorder characterized by defective intracellular killing of microorganisms due to the neutrophils' inability to generate superoxide ions. Although it is always caused by mutations in the CYBB gene, clinical and molecular characteristics vary in different ethnic backgrounds. Two unrelated Thai boys presented with severe persistent pulmonary infections at the age of two months. Their abnormal dihydrorhodamine (DHR) flow cytometry assays supported the diagnosis of X-CGD. Mutation analysis was performed by polymerase chain reaction (PCR) amplification and sequencing of the entire coding regions of CYBB. Mutations identified were confirmed by restriction enzyme analyses. PCR-sequencing of the entire coding regions of CYBB identified nonsense mutations, 271C>T (R91X) in exon 4 and 456T>A (Y152X) in exon 5, in probands of each family. Both of the patients' mothers were found to be carriers. This observation supports that CYBB is the gene responsible for X-CGD across different populations and nonsense mutations are associated with severe phenotypes.
Subject(s)
Adult , Codon, Nonsense , DNA Mutational Analysis , Exons , Female , Granulomatous Disease, Chronic/genetics , Heterozygote , Homozygote , Humans , Infant , Male , Membrane Glycoproteins/genetics , Mothers , NADPH Oxidases/genetics , Polymerase Chain Reaction , ThailandABSTRACT
We evaluated a boy who had multiple Salmonella septicemia, Aspergillus pneumonia and brain abscesses. His nitroblue tetrazolium (NBT) test was reportedly abnormal. The dihydrorhodamine (DHR) flow cytometry assay was compatible with typical X-linked chronic granulomatous disease (X-CGD). CYBB analysis revealed a novel complex mutation atggacg --> ttca in exon 12 (base pairs 1532-1538). As a result, 3 amino acids Tyr 511, Gly 512 and Arg 513 were deleted and replaced by 2 amino acids, Phe and Gln. The DHR and mutation analysis of his mother showed normal DHR pattern and no mutations in exon 12 of CYBB gene. In conclusion, any children with multiple Salmonella and Aspergillus infection should be suspected of CGD. NBT test, DHR assay and gene analysis are helpful toolsto confirm the diagnosis e v en i n the case of de novo mutation.
Subject(s)
Amino Acid Sequence , Amino Acid Substitution , Aspergillosis, Allergic Bronchopulmonary/complications , Granulomatous Disease, Chronic/complications , Humans , Infant , Male , Membrane Glycoproteins/genetics , NADPH Oxidases/genetics , Pneumonia/complications , Salmonella Infections/complications , Sepsis/complications , Sequence DeletionABSTRACT
Background: The X-linked form of chronic granulomatous disease (CGD) is a primary immunodeficiency that affects phagocytes of the innate immune system and is characterized by an increased susceptibility to severe bacterial and fungal infections. It is caused by mutations in the CYBB gene, which encodes the 91-kD subunit of phagocyte NADPH oxidase. Aim: To identify the mutation in the CYBB gene in two unrelated patients from Chile with the diagnosis of X-linked CGD and their families. Patients and methods: The molecular genetic defects of two unrelated patients from Chile with X-linked CGD caused by defects in the CYBB gene were investigated. The underlying mutation was investigated by single strand conformation polymorphism (SSCP) analysis of PCR-amplified genomic DNA and by sequencing of the affected gene region. Results: We found an insertion c.1267_1268insA in exon 10 leading to a frameshift mutation. This mutation is a novel report. We also identified a splice site mutation in the other patient, that presented a c.1326 +1 G>A substitution in intron 10. The mutation was also detectable in his heterozygous mother. Conclusions: This is the first report of the clinical and molecular characterization of Chilean patients with mutations in CYBB gene.
Subject(s)
Child , Child, Preschool , Humans , Male , Frameshift Mutation/genetics , Granulomatous Disease, Chronic/genetics , Membrane Glycoproteins/genetics , Mutagenesis, Insertional/genetics , NADPH Oxidases/genetics , Case-Control Studies , Chile , Granulomatous Disease, Chronic/diagnosis , RNA Splice Sites , Sequence Analysis, DNAABSTRACT
Background: The cytosolic protein p47-phox (phagocyte oxidase) is one of the essential components of the superoxide generating system in phagocytes and its defect causes approximately 30 percent of the chronic granulomatous disease (CGD) cases. Aim: Two patients were studied, belonging to the same family, without a consanguinous background, in which deficiency or absence of superoxide generation was found together with recurrent and severe infections in one case and benign infections in the second. Methods: The presence of gp91-, p67- and p47-phox in patients and controls was determined by Western Blot analysis of granulocytes. Sequencing of PCR amplified DNA was performed by an enzimatic method. Results: Western Blot analysis showed normal expression of gp91 and p67 and absence of p47-phox. The molecular genetic study demonstrated a homocygotic dinucleotide GT (GT) deletion at the beginning of exon 2 of the p47-phox gene. The same mutation has been found in European, American and Japanese patients. Conclusions: The molecular characterization of this pathology done for the first time in Chile is important for diagnostic classification, patient prognosis, and adequate genetic advice and a possible future therapy