RESUMEN
Diabetic retinopathy (DR) is a complication of diabetes with a complex pathophysiology and multiple factors involved. Recently, it has been found that the upregulation of the renin-angiotensin-aldosterone system (RAAS) leads to overexpression of angiotensin II (Ang II), which induces oxidative stress, inflammation, and angiogenesis in the retina. Therefore, RAAS may be a promising therapeutic target in DR. Notably, RAAS inhibitors are often used in the treatment of hypertension. Still, the potential role and mechanism of DR must be further studied. In this review, we discuss and summarize the pathology and potential therapeutic goals of RAAS in DR.
Asunto(s)
Retinopatía Diabética , Sistema Renina-Angiotensina , Humanos , Retinopatía Diabética/tratamiento farmacológico , Retinopatía Diabética/fisiopatología , Sistema Renina-Angiotensina/fisiología , Sistema Renina-Angiotensina/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/fisiología , Inhibidores de la Enzima Convertidora de Angiotensina/uso terapéutico , Angiotensina II/fisiología , AnimalesRESUMEN
ABSTRACT Diabetic retinopathy (DR) is a complication of diabetes with a complex pathophysiology and multiple factors involved. Recently, it has been found that the upregulation of the renin-angiotensin-aldosterone system (RAAS) leads to overexpression of angiotensin II (Ang II), which induces oxidative stress, inflammation, and angiogenesis in the retina. Therefore, RAAS may be a promising therapeutic target in DR. Notably, RAAS inhibitors are often used in the treatment of hypertension. Still, the potential role and mechanism of DR must be further studied. In this review, we discuss and summarize the pathology and potential therapeutic goals of RAAS in DR.
RESUMEN
OBJECTIVES: To investigate the cardiovascular features and endothelium in neonates born to mothers with preeclampsia. STUDY DESIGN: In this combined observational cohort and case-control study, neonates born to mothers with normotension and mothers with preeclampsia were recruited at a neonatal intensive care unit of a tertiary medical center. Cardiovascular measurements by echocardiography and the clinical measures upon admission were analyzed. Vascular cell adhesion molecule-1 expression in umbilical arteries and in in vitro endothelial cell stimulation with plasma were examined. Continuous data were compared using nonparametric analysis, and their relationships were analyzed using linear regression. Binary logistic regression was performed in the model of adjustment of birth body weight and for multivariate analysis. RESULTS: In the cohort, almost all cardiovascular segments positively correlated to birth weight. Notably, neonates (n = 65) of mothers with preeclampsia had significantly larger coronary arteries at birth than neonates of mothers with normotension (n = 404) (median size of left main coronary artery 1.36 mm versus 1.08 mm, p <0.001; median size of right coronary artery, RCA 1.25 mm versus 1.0 mm, p <0.001). The size of the right coronary artery positively correlated to the maternal antepartum diastolic blood pressure (r = 0.298, P = .018) and was associated with in-hospital death (P < .001). Meanwhile, endothelial vascular cell adhesion molecule-1 expression was significantly increased in the umbilical arteries of the preeclamptic group and following preeclamptic cord-plasma stimulation. The latter also correlated with their relative coronary sizes. CONCLUSIONS: Neonates of mothers with preeclampsia had distinctive coronary dilatation at birth. Coronary size might be useful as a severity index of neonatal endothelial inflammation as a result of maternal preeclampsia.
Asunto(s)
Enfermedad de la Arteria Coronaria/etiología , Vasos Coronarios/fisiopatología , Endotelio Vascular/fisiopatología , Inflamación/diagnóstico , Preeclampsia/diagnóstico , Enfermedad de la Arteria Coronaria/diagnóstico , Enfermedad de la Arteria Coronaria/fisiopatología , Dilatación Patológica/diagnóstico , Dilatación Patológica/etiología , Dilatación Patológica/fisiopatología , Endotelio Vascular/diagnóstico por imagen , Femenino , Estudios de Seguimiento , Edad Gestacional , Humanos , Recién Nacido , Inflamación/fisiopatología , Masculino , Embarazo , Estudios RetrospectivosRESUMEN
INTRODUCTION: Community-acquired pneumonia (CAP) is a global disease responsible for a large number of deaths, with significant economic impact. As diagnostic tools have increased in sensitivity, understanding of the etiology of CAP has begun to change. Mycoplasma pneumoniae is one of the major pathogens causing CAP. Macrolides and related antibiotics are first-line treatments for M. pneumoniae. Macrolide resistance has been spreading for 15 years and now occurs in worldwide. We undertook the first study on macrolide resistance of M. pneumoniae in Yantai. This may be helpful to determine the appropriate therapy for CAP in this population. OBJECTIVE: To investigate the rate and mechanism of macrolide resistance in Yantai. METHODS: Pharyngeal swab samples were collected from adult CAP patients. Samples were assayed by polymerase chain reaction (PCR) and cultivated to test for M. pneumoniae. Nested PCR was used to specifically amplify M. pneumoniae 23S rRNA gene fragments containing mutations, and amplicons were analyzed by CE-SSCP for macrolide resistance mutations. Results were confirmed by sequencing. Twenty-seven strains of M. pneumoniae were isolated and the activities of nine antibiotics against M. pneumoniae were tested in vitro. RESULTS: Out of 128 samples tested, 27 were positive for M. pneumoniae. Mycoplasma 100% macrolides resistance to Mycoplasma pneumoniae. The mechanism of macrolides resistance was A2063G point mutation in the sequence directly binding to macrolides in the 23S rRNA V domain in vitro. The mean pyretolytic time for the fluoroquinolone group was 4.7 ±2.9 d, which was significantly shorter than 8.2 ±4.1 d for the azithromycin group. CONCLUSIONS: Macrolides are not the first-line treatment for M. pneumoniae respiratory tract infections in Yantai.
INTRODUCCIÓN: Neumonía adquirida por en la comunidad (NAC) es una enfermedad responsable por un gran número de muertes y un impacto económico importante. Debido a que el diagnostico incrementó la sensibilidad, se cambió la etiología de la NAC. Adicionalmente, Mycoplasma pneumoniae es uno de los patógenos que causan la NAC. Los macrólidos y antibióticos relacionados son la primera línea de tratamiento para M. pneumoniae. La resistencia a macrólidos se aumentó en los últimos 15 años y ahora se encuentra distribuido en todo el mundo. Nosotros realizamos el primer estudio de resitencia a M. pneumoniae a los macrólidos en Yantai. Esto podría ser útil para determinar una terapia apropiada para NAC en esta población. OBJETIVO: Investigar la tasa y el mecanismo para la resitencia a los macrólidos en Yantai. MÉTODOS: Se colectaron muestras faringeas usando un hisopo. Las muestras se analizaron mediante la reacción en cadena de la polimerasa (PCR) y por cultivo para M. pneumoniae. Se uso una PCR anidad para amplificar fragmentos del gen 23S rRNA especifico con las mutaciones para M. pneumoniae. Se analizaron amplicomes por CE-SSCP para determinar la resitencia a los macrólidos. Estos resultados se confirmaron por secuenciación. Se aislaron 27 cepas de M. pneumoniae y se probaron nueve antibióticos in vitro. RESULTADOS: De 128 muestras, 27 fueron positivas para M. pneumoniae. Se determinó una resistencia a macrólidos por Mycoplasma del 100%. Los mecanismos de esta resitencia fue una mutacion punctual A2063G en la secuencia que se une directamente a los macrólidos en el dominio 23S rRNA V in vitro. El tiempo piotolítico medio para el grupo de fluoroquinolonas fue 4.7 ±2.9 d, que fue significativamente más corto que para el grupo de azitromicina: 8.2 ±4.1 d. CONCLUSIONES: Los macrólidos no son la primera linea de tratamiento para las infecciones del tracto respiratorio contra M. pneumoniae respiratory tract infections en Yantai.
Asunto(s)
Antibacterianos/farmacología , Infecciones Comunitarias Adquiridas/epidemiología , Mycoplasma pneumoniae/aislamiento & purificación , Neumonía por Mycoplasma/epidemiología , Adolescente , Adulto , Anciano , China/epidemiología , Infecciones Comunitarias Adquiridas/tratamiento farmacológico , Infecciones Comunitarias Adquiridas/microbiología , Farmacorresistencia Bacteriana/genética , Femenino , Humanos , Macrólidos/farmacología , Masculino , Persona de Mediana Edad , Neumonía por Mycoplasma/tratamiento farmacológico , Neumonía por Mycoplasma/microbiología , Mutación Puntual , Reacción en Cadena de la Polimerasa , Adulto JovenRESUMEN
OBJECTIVE: To evaluate the impact of parental weight status and offspring cardiorespiratory fitness on the risk of obesity among Chinese children. STUDY DESIGN: This cross-sectional study was conducted in Wuhan, China from May to June 2010. Children's height, weight, and waist circumference were measured for assessing their total and central obesity. Their cardiorespiratory fitness was determined by the 20-m shuttle-run test. We calculated parental body mass index according to self-reported height and weight, and divided it into normal weight or overweight/obesity. Multivariable logistic regression model was applied to estimate the combined relationships of cardiorespiratory fitness and parental weight status with the risk of obesity of children. RESULTS: A total of 587 Chinese children (343 boys and 244 girls) aged 9.6 (0.7) years participated in this study. Compared with those who had low cardiorespiratory fitness and at least 1 parent with overweight/obesity, children who had high cardiorespiratory fitness and at least 1 parent with overweight/obesity reported lower risks of total obesity (OR 0.12, 95% CI .05-0.30) and central obesity (OR .09, 95% CI .04-0.20), and children who had high cardiorespiratory fitness and no parent with overweight/obesity were 89% (OR 0.11, 95% CI .05-0.24) less likely to have total obesity and 92% (OR .08, 95% CI .04-0.16) less likely to have central obesity (all P < .001). CONCLUSIONS: High level of cardiorespiratory fitness among children could attenuate the influence of parental obesity on their offspring's weight status.
Asunto(s)
Capacidad Cardiovascular/fisiología , Sobrepeso/prevención & control , Padres , Obesidad Infantil/prevención & control , Índice de Masa Corporal , Niño , China/epidemiología , Estudios Transversales , Femenino , Estudios de Seguimiento , Humanos , Incidencia , Masculino , Sobrepeso/epidemiología , Sobrepeso/fisiopatología , Obesidad Infantil/epidemiología , Obesidad Infantil/fisiopatología , Estudios Retrospectivos , Factores de RiesgoRESUMEN
Abstract Introduction: Community-acquired pneumonia (CAP) is a global disease responsible for a large number of deaths, with significant economic impact. As diagnostic tools have increased in sensitivity, understanding of the etiology of CAP has begun to change. Mycoplasma pneumoniae is one of the major pathogens causing CAP. Macrolides and related antibiotics are first-line treatments for M. pneumoniae. Macrolide resistance has been spreading for 15 years and now occurs in worldwide. We undertook the first study on macrolide resistance of M. pneumoniae in Yantai. This may be helpful to determine the appropriate therapy for CAP in this population. Objective: To investigate the rate and mechanism of macrolide resistance in Yantai. Methods: Pharyngeal swab samples were collected from adult CAP patients. Samples were assayed by polymerase chain reaction (PCR) and cultivated to test for M. pneumoniae. Nested PCR was used to specifically amplify M. pneumoniae 23S rRNA gene fragments containing mutations, and amplicons were analyzed by CE-SSCP for macrolide resistance mutations. Results were confirmed by sequencing. Twenty-seven strains of M. pneumoniae were isolated and the activities of nine antibiotics against M. pneumoniae were tested in vitro. Results: Out of 128 samples tested, 27 were positive for M. pneumoniae. Mycoplasma 100% macrolides resistance to Mycoplasma pneumoniae. The mechanism of macrolides resistance was A2063G point mutation in the sequence directly binding to macrolides in the 23S rRNA V domain in vitro. The mean pyretolytic time for the fluoroquinolone group was 4.7 ±2.9 d, which was significantly shorter than 8.2 ±4.1 d for the azithromycin group. Conclusions: Macrolides are not the first-line treatment for M. pneumoniae respiratory tract infections in Yantai.
Resumen Introducción: Neumonía adquirida por en la comunidad (NAC) es una enfermedad responsable por un gran número de muertes y un impacto económico importante. Debido a que el diagnostico incrementó la sensibilidad, se cambió la etiología de la NAC. Adicionalmente, Mycoplasma pneumoniae es uno de los patógenos que causan la NAC. Los macrólidos y antibióticos relacionados son la primera línea de tratamiento para M. pneumoniae. La resistencia a macrólidos se aumentó en los últimos 15 años y ahora se encuentra distribuido en todo el mundo. Nosotros realizamos el primer estudio de resitencia a M. pneumoniae a los macrólidos en Yantai. Esto podría ser útil para determinar una terapia apropiada para NAC en esta población. Objetivo: Investigar la tasa y el mecanismo para la resitencia a los macrólidos en Yantai. Métodos: Se colectaron muestras faringeas usando un hisopo. Las muestras se analizaron mediante la reacción en cadena de la polimerasa (PCR) y por cultivo para M. pneumoniae. Se uso una PCR anidad para amplificar fragmentos del gen 23S rRNA especifico con las mutaciones para M. pneumoniae. Se analizaron amplicomes por CE-SSCP para determinar la resitencia a los macrólidos. Estos resultados se confirmaron por secuenciación. Se aislaron 27 cepas de M. pneumoniae y se probaron nueve antibióticos in vitro. Resultados: De 128 muestras, 27 fueron positivas para M. pneumoniae. Se determinó una resistencia a macrólidos por Mycoplasma del 100%. Los mecanismos de esta resitencia fue una mutacion punctual A2063G en la secuencia que se une directamente a los macrólidos en el dominio 23S rRNA V in vitro. El tiempo piotolítico medio para el grupo de fluoroquinolonas fue 4.7 ±2.9 d, que fue significativamente más corto que para el grupo de azitromicina: 8.2 ±4.1 d. Conclusiones: Los macrólidos no son la primera linea de tratamiento para las infecciones del tracto respiratorio contra M. pneumoniae respiratory tract infections en Yantai.
Asunto(s)
Adolescente , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Neumonía por Mycoplasma/epidemiología , Infecciones Comunitarias Adquiridas/epidemiología , Antibacterianos/farmacología , Mycoplasma pneumoniae/aislamiento & purificación , Neumonía por Mycoplasma/microbiología , Neumonía por Mycoplasma/tratamiento farmacológico , China/epidemiología , Reacción en Cadena de la Polimerasa , Mutación Puntual , Infecciones Comunitarias Adquiridas/microbiología , Infecciones Comunitarias Adquiridas/tratamiento farmacológico , Macrólidos/farmacología , Farmacorresistencia Bacteriana/genéticaRESUMEN
ABSTRACT To seek a simple, rapid and sensitive Coprinus cinereus Peroxidase (CIP) activity assay, a convenient one-factor-at-a-time (OFAT) method and a response surface methodology (RSM) were used. The recombinant CIP expressed in Pichia pastoris was purified with the Ni-NTA spin column. Based on the results of catalytic efficiency (kcat/Km) analysis, 2,2'-azinobis (ethylbenzthiazoline -6-sulfonate) (ABTS) was selected as the optimal enzyme substrate. Results of the OFAT method showed that enzymatic reaction performed in 0.1 mol/L sodium acetate (pH 5.0) buffer in a 200-µl reaction mixture containing 0.5 mmol/L ABTS, 10 mmol/L hydrogen peroxide (H2O2), 49.7 ng CIP at 25°C gave an average CIP activity of 88 U/mL. The ABTS and H2O2 concentrations were then further optimized to improve the sensitivity of the assay. To do that, RSM was conducted through central composite design, and a reduced quadratic model with good fit regression equation was generated. ANOVA analysis of this model indicated that the concentrations of ABTS and H2O2 and their interaction had significant impact on the assay sensitivity. The optimal reaction mixture was determined to include an initial ABTS concentration of 0.82 mmol/L 49.7 ng CIP and 16.36 mmol/L H2O2, and the activity under this condition was determined to be 138.89 U/mL.
RESUMEN
Abstract Background The mechanism underlying the coexistence of hepatitis B surface antigen and antibodies to HBsAg in chronic hepatitis B patients remains unknown. Aims This research aimed to determine the clinical and virological features of the rare pattern. Methods A total of 32 chronic hepatitis B patients infected by HBV genotype C were included: 15 carrying both HBsAg and anti-HBs (group I) and 17 solely positive for HBsAg (group II). S gene and reverse transcriptase region sequences were amplified, sequenced and compared with the reference sequences. Results The amino acid variability within major hydrophilic region, especially the “a” determinant region, and within reverse transcriptase for regions overlapping the major hydrophilic region in group I is significantly higher than those in group II. Mutation sI126S/T within the “a” determinant was the most frequent change, and only patients from group I had the sQ129R, sG130N, sF134I, sG145R amino acid changes, which are known to alter immunogenicity. Conclusions In chronic patients, the concurrent HBsAg/anti-HBs serological profile is associated with an increased aa variability in several key areas of HBV genome. Additional research on these genetic mutants are needed to clarify their biological significance for viral persistence.
Asunto(s)
Adulto , Femenino , Humanos , Masculino , Adulto Joven , Anticuerpos contra la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/sangre , Virus de la Hepatitis B/genética , Hepatitis B Crónica/genética , Hepatitis B Crónica/inmunología , ADN Polimerasa Dirigida por ARN/genética , Proteínas del Envoltorio Viral/genética , China , ADN Viral , Genotipo , Virus de la Hepatitis B/inmunología , Mutación , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADNRESUMEN
BACKGROUND: The mechanism underlying the coexistence of hepatitis B surface antigen and antibodies to HBsAg in chronic hepatitis B patients remains unknown. AIMS: This research aimed to determine the clinical and virological features of the rare pattern. METHODS: A total of 32 chronic hepatitis B patients infected by HBV genotype C were included: 15 carrying both HBsAg and anti-HBs (group I) and 17 solely positive for HBsAg (group II). S gene and reverse transcriptase region sequences were amplified, sequenced and compared with the reference sequences. RESULTS: The amino acid variability within major hydrophilic region, especially the "a" determinant region, and within reverse transcriptase for regions overlapping the major hydrophilic region in group I is significantly higher than those in group II. Mutation sI126S/T within the "a" determinant was the most frequent change, and only patients from group I had the sQ129R, sG130N, sF134I, sG145R amino acid changes, which are known to alter immunogenicity. CONCLUSIONS: In chronic patients, the concurrent HBsAg/anti-HBs serological profile is associated with an increased aa variability in several key areas of HBV genome. Additional research on these genetic mutants are needed to clarify their biological significance for viral persistence.
Asunto(s)
Anticuerpos contra la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/sangre , Virus de la Hepatitis B/genética , Hepatitis B Crónica/genética , Hepatitis B Crónica/inmunología , ADN Polimerasa Dirigida por ARN/genética , Proteínas del Envoltorio Viral/genética , Adulto , China , ADN Viral , Femenino , Genotipo , Virus de la Hepatitis B/inmunología , Humanos , Masculino , Mutación , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , Adulto JovenRESUMEN
Multidrug resistance (MDR) poses a serious impediment to the success of chemotherapy for laryngeal cancer. To identify microRNAs and mRNAs associated with MDR of human laryngeal cancer Hep-2 cells, we developed a multidrug-resistant human laryngeal cancer subline, designated Hep-2/v, by exposing Hep-2 cells to stepwise increasing concentrations of vincristine (0.02-0.96'µM). Microarray assays were performed to compare the microRNA and mRNA expression profiles of Hep-2 and Hep-2/v cells. Compared to Hep-2 cells, Hep-2/v cells were more resistant to chemotherapy drugs (∼45-fold more resistant to vincristine, 5.1-fold more resistant to cisplatin, and 5.6-fold more resistant to 5-fluorouracil) and had a longer doubling time (42.33±1.76 vs 28.75±1.12'h, P<0.05), higher percentage of cells in G0/G1 phase (80.98±0.52 vs 69.14±0.89, P<0.05), increased efflux of rhodamine 123 (95.97±0.56 vs 12.40±0.44%, P<0.01), and up-regulated MDR1 expression. A total of 7 microRNAs and 605 mRNAs were differentially expressed between the two cell types. Of the differentially expressed mRNAs identified, regulator of G-protein signaling 10, high-temperature requirement protein A1, and nuclear protein 1 were found to be the putative targets of the differentially expressed microRNAs identified. These findings may open a new avenue for clarifying the mechanisms responsible for MDR in laryngeal cancer.
Asunto(s)
Humanos , Resistencia a Antineoplásicos/genética , Neoplasias Laríngeas/genética , MicroARNs/aislamiento & purificación , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , ARN Mensajero/aislamiento & purificación , Antineoplásicos/farmacología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cisplatino/farmacología , Citometría de Flujo , Fluorouracilo/farmacología , Puntos de Control de la Fase G1 del Ciclo Celular , Genes MDR , Neoplasias Laríngeas/tratamiento farmacológico , Proteínas de Neoplasias/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteínas RGS/genética , /farmacocinética , Serina Endopeptidasas/genética , Análisis de Matrices Tisulares , Vincristina/farmacologíaRESUMEN
Multidrug resistance (MDR) poses a serious impediment to the success of chemotherapy for laryngeal cancer. To identify microRNAs and mRNAs associated with MDR of human laryngeal cancer Hep-2 cells, we developed a multidrug-resistant human laryngeal cancer subline, designated Hep-2/v, by exposing Hep-2 cells to stepwise increasing concentrations of vincristine (0.02-0.96'µM). Microarray assays were performed to compare the microRNA and mRNA expression profiles of Hep-2 and Hep-2/v cells. Compared to Hep-2 cells, Hep-2/v cells were more resistant to chemotherapy drugs (≈ 45-fold more resistant to vincristine, 5.1-fold more resistant to cisplatin, and 5.6-fold more resistant to 5-fluorouracil) and had a longer doubling time (42.33 ± 1.76 vs 28.75 ± 1.12'h, P<0.05), higher percentage of cells in G0/G1 phase (80.98 ± 0.52 vs 69.14 ± 0.89, P<0.05), increased efflux of rhodamine 123 (95.97 ± 0.56 vs 12.40 ± 0.44%, P<0.01), and up-regulated MDR1 expression. A total of 7 microRNAs and 605 mRNAs were differentially expressed between the two cell types. Of the differentially expressed mRNAs identified, regulator of G-protein signaling 10, high-temperature requirement protein A1, and nuclear protein 1 were found to be the putative targets of the differentially expressed microRNAs identified. These findings may open a new avenue for clarifying the mechanisms responsible for MDR in laryngeal cancer.
Asunto(s)
Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , Resistencia a Antineoplásicos/genética , Neoplasias Laríngeas/genética , MicroARNs/aislamiento & purificación , ARN Mensajero/aislamiento & purificación , Subfamilia B de Transportador de Casetes de Unión a ATP , Antineoplásicos/farmacología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cisplatino/farmacología , Citometría de Flujo , Fluorouracilo/farmacología , Puntos de Control de la Fase G1 del Ciclo Celular , Genes MDR , Serina Peptidasa A1 que Requiere Temperaturas Altas , Humanos , Neoplasias Laríngeas/tratamiento farmacológico , Proteínas de Neoplasias/genética , Proteínas RGS/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Rodamina 123/farmacocinética , Serina Endopeptidasas/genética , Análisis de Matrices Tisulares , Vincristina/farmacologíaRESUMEN
OBJECTIVE: This study aimed to identify novel GATA5 mutations that underlie familial atrial fibrillation. METHODS: A total of 110 unrelated patients with familial atrial fibrillation and 200 unrelated, ethnically matched healthy controls were recruited. The entire coding region of the GATA5 gene was sequenced in 110 atrial fibrillation probands. The available relatives of the mutation carriers and 200 controls were subsequently genotyped for the identified mutations. The functional effect of the mutated GATA5 was characterized using a luciferase reporter assay system. RESULTS: Two novel heterozygous GATA5 mutations (p.Y138F and p.C210G) were identified in two of the 110 unrelated atrial fibrillation families. These missense mutations cosegregated with AF in the families and were absent in the 400 control chromosomes. A cross-species alignment of GATA5 protein sequence showed that the altered amino acids were completely conserved evolutionarily. A functional analysis revealed that the mutant GATA5 proteins were associated with significantly decreased transcriptional activation when compared with their wild-type counterpart. CONCLUSION: The findings expand the spectrum of GATA5 mutations linked to AF and provide novel insights into the molecular mechanism involved in the pathogenesis of atrial fibrillation, suggesting potential implications for the early prophylaxis and personalized treatment of this common arrhythmia.
Asunto(s)
Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Fibrilación Atrial/genética , /genética , Mutación Missense/genética , Secuencia de Aminoácidos , Pueblo Asiatico/genética , Fibrilación Atrial/etnología , Estudios de Casos y Controles , Distribución de Chi-Cuadrado , Análisis Mutacional de ADN , Heterocigoto , Luciferasas/genética , Linaje , Alineación de SecuenciaRESUMEN
Background: Bacterial wilt caused by Ralstonia solanacearum is the most devastating disease in peanut. Planting resistant peanut cultivars is deemed as the sole economically viable means for effective control of the disease. To understand the molecular mechanism underlying resistance and facilitate breeding process, differences in gene expression between seeds of Rihua 1 (a Virginia type peanut variety resistant to bacterial wilt) inoculated with the bacterial pathogen suspension (10(9) cfu ml-1) and seeds of the same cultivar treated with water (control), were studied using the GenefishingTM technology. Results: A total of 25 differentially expressed genes were isolated. Expression of genes encoding cyclophilin and ADP-ribosylation factor, respectively, were further studied by real time RT-PCR, and full length cDNAs of both genes were obtained by rapid amplification of cDNA ends. Conclusions: The study provided candidate genes potentially useful for breeding peanut cultivars with both high yield and bacterial wilt resistance, although confirmation of their functions through transgenic studies is still needed.
Asunto(s)
Arachis/genética , Factores de Ribosilacion-ADP/genética , Ralstonia solanacearum/patogenicidad , Inmunidad Innata , Reacción en Cadena en Tiempo Real de la Polimerasa , Análisis de SecuenciaRESUMEN
OBJECTIVE: This study aimed to identify novel GATA5 mutations that underlie familial atrial fibrillation. METHODS: A total of 110 unrelated patients with familial atrial fibrillation and 200 unrelated, ethnically matched healthy controls were recruited. The entire coding region of the GATA5 gene was sequenced in 110 atrial fibrillation probands. The available relatives of the mutation carriers and 200 controls were subsequently genotyped for the identified mutations. The functional effect of the mutated GATA5 was characterized using a luciferase reporter assay system. RESULTS: Two novel heterozygous GATA5 mutations (p.Y138F and p.C210G) were identified in two of the 110 unrelated atrial fibrillation families. These missense mutations cosegregated with AF in the families and were absent in the 400 control chromosomes. A cross-species alignment of GATA5 protein sequence showed that the altered amino acids were completely conserved evolutionarily. A functional analysis revealed that the mutant GATA5 proteins were associated with significantly decreased transcriptional activation when compared with their wild-type counterpart. CONCLUSION: The findings expand the spectrum of GATA5 mutations linked to AF and provide novel insights into the molecular mechanism involved in the pathogenesis of atrial fibrillation, suggesting potential implications for the early prophylaxis and personalized treatment of this common arrhythmia.
Asunto(s)
Fibrilación Atrial/genética , Factor de Transcripción GATA5/genética , Mutación Missense/genética , Adulto , Secuencia de Aminoácidos , Pueblo Asiatico/genética , Fibrilación Atrial/etnología , Estudios de Casos y Controles , Distribución de Chi-Cuadrado , Análisis Mutacional de ADN , Femenino , Heterocigoto , Humanos , Luciferasas/genética , Masculino , Persona de Mediana Edad , Linaje , Alineación de Secuencia , Adulto JovenRESUMEN
Antibacterial monomers incorporated in dentin bonding systems may have toxic effects on the pulp. Thus, the cytotoxicity of antibacterial monomers and its underlying mechanisms must be elucidated to improve the safety of antibacterial monomer application. The influence of an antibacterial monomer, methacryloxylethyl cetyl ammonium chloride (DMAE-CB), on the vitality of L929 mouse fibroblasts was tested using MTT assay. Cell cycle progression was studied using flow cytometry. Production of intracellular reactive oxygen species (ROS) after DMAE-CB treatment was measured using 2,7-dichlorodihydrofluorescein diacetate staining and flow cytometry analysis. Loss of mitochondrial membrane potential, disturbance of Bcl-2 and Bax expression, as well as release of cytochrome C were also measured using flow cytometry analysis or Western blot to explore the possible involvement of the mitochondrial-related apoptotic pathway. DMAE-CB elicited cell death in a dose-dependent manner and more than 50 percent of cells were killed after treatment with 30 µM of the monomer. Both necrosis and apoptosis were observed. DMAE-CB also induced G1- and G2-phase arrest. Increased levels of intracellular ROS were observed after 1 h and this overproduction was further enhanced by 6-h treatment with the monomer. DMAE-CB may cause apoptosis by disturbing the expression of Bcl-2 and Bax, reducing the mitochondrial potential and inducing release of cytochrome C. Taken together, these findings suggest that the toxicity of the antibacterial monomer DMAE-CB is associated with ROS production, mitochondrial dysfunction, cell cycle disturbance, and cell apoptosis/necrosis.
Asunto(s)
Animales , Ratones , Antibacterianos/toxicidad , Apoptosis/efectos de los fármacos , Recubrimientos Dentinarios/toxicidad , Metacrilatos/toxicidad , Mitocondrias/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Compuestos de Amonio Cuaternario/toxicidad , Análisis de Varianza , Fibroblastos/efectos de los fármacos , Modelos Animales , Especies Reactivas de Oxígeno/metabolismo , Estadísticas no ParamétricasRESUMEN
Antibacterial monomers incorporated in dentin bonding systems may have toxic effects on the pulp. Thus, the cytotoxicity of antibacterial monomers and its underlying mechanisms must be elucidated to improve the safety of antibacterial monomer application. The influence of an antibacterial monomer, methacryloxylethyl cetyl ammonium chloride (DMAE-CB), on the vitality of L929 mouse fibroblasts was tested using MTT assay. Cell cycle progression was studied using flow cytometry. Production of intracellular reactive oxygen species (ROS) after DMAE-CB treatment was measured using 2,7-dichlorodihydrofluorescein diacetate staining and flow cytometry analysis. Loss of mitochondrial membrane potential, disturbance of Bcl-2 and Bax expression, as well as release of cytochrome C were also measured using flow cytometry analysis or Western blot to explore the possible involvement of the mitochondrial-related apoptotic pathway. DMAE-CB elicited cell death in a dose-dependent manner and more than 50% of cells were killed after treatment with 30 µM of the monomer. Both necrosis and apoptosis were observed. DMAE-CB also induced G1- and G2-phase arrest. Increased levels of intracellular ROS were observed after 1 h and this overproduction was further enhanced by 6-h treatment with the monomer. DMAE-CB may cause apoptosis by disturbing the expression of Bcl-2 and Bax, reducing the mitochondrial potential and inducing release of cytochrome C. Taken together, these findings suggest that the toxicity of the antibacterial monomer DMAE-CB is associated with ROS production, mitochondrial dysfunction, cell cycle disturbance, and cell apoptosis/necrosis.
Asunto(s)
Antibacterianos/toxicidad , Apoptosis/efectos de los fármacos , Recubrimientos Dentinarios/toxicidad , Metacrilatos/toxicidad , Mitocondrias/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Compuestos de Amonio Cuaternario/toxicidad , Análisis de Varianza , Animales , Fibroblastos/efectos de los fármacos , Ratones , Modelos Animales , Especies Reactivas de Oxígeno/metabolismo , Estadísticas no ParamétricasRESUMEN
Internalization of Staphylococcus aureus in bovine endothelial cells (BEC) is increased by tumor necrosis factor alpha stimulation and NF-κB activation. Because the phosphoinositide-3-kinase (PI3K)-Akt signaling pathway also modulates NF-κB activity, we considered whether the internalization of S. aureus by BEC is associated with the activity of PI3K and Akt. We found a time- and multiplicity of infection-dependent phosphorylation of Akt on Ser473 in BEC infected with S. aureus. This phosphorylation was inhibited by LY294002 (LY), indicating the participation of PI3K. Inhibition of either PI3K with LY or wortmannin, or Akt with SH-5, strongly reduced the internalization of S. aureus. Transfection of BEC with a dominant-negative form of the Akt gene significantly decreased S. aureus internalization, whereas transfection with the constitutively active mutant increased the number of internalized bacterium. Inhibition of PDK1 activity with OSU-03012 did not affect the level of S. aureus internalization, demonstrating that phosphorylation of Akt on Thr308 is not important for this process. Compared to the untreated control, the adherence of S. aureus to the surface of BEC was unaltered when cells were transfected or incubated with the pharmacological inhibitors. Furthermore, Akt activation by internalized S. aureus triggered a time-dependent phosphorylation of glycogen synthase kinase-3α (GSK-3α) on Ser21 and GSK-3ß on Ser9 that was partially inhibited with SH-5. Finally, treatment of BEC with LY prior to S. aureus infection inhibited the NF-κB p65 subunit phosphorylation on Ser536, indicating the involvement of PI3K. These results suggest that PI3K-Akt activity is important for the internalization of S. aureus and phosphorylation of GSK-3α, GSK-3ß, and NF-κB.