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AIM: This study aimed to evaluate the association of toll-like receptor (TLR) inflammatory cascade with the development of diabetic kidney disease (DKD) in children and adolescents with type 1 diabetes (T1D). METHODS: A total of 49 T1D patients and 49 normoglycaemic (NG) subjects aged 5-20 years old were recruited. TLR2, TLR4, MYD88, NFKB, MCP1/CCL2 and IL18 mRNA expressions were measured in peripheral blood mononuclear cells by reverse transcription-quantitative polymerase chain reaction. Fasting glucose, glycated haemoglobin, serum urea, serum creatinine and urinary albumin-to-creatinine ratio (ACR) were determined. RESULTS: The mRNA expressions of TLR2, TLR4, MYD88 and NFKB were significantly increased in the T1D group compared with the NG group. The mRNA expression levels of MCP1/CCL2 and IL18 were higher in 21 T1D patients (42.9%) (average of MCP1/CCL2: 6.6-fold and IL18: 5.8-fold) than in NG patients. Furthermore, ACR was increased in the T1D group compared with the NG group. CONCLUSION: The increased mRNA expression of TLR2, TLR4, MYD88, NFKB, MCP1/CCL2 and IL18 favours the development of an inflammatory process that may lead to a decline in renal function and consequently DKD in children and adolescents with T1D. This suggests that these genes are early mediators of onset DKD since the beginning of the lives of the paediatric T1D patients.
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Diabetes Mellitus Tipo 1 , Nefropatías Diabéticas , Adolescente , Adulto , Niño , Preescolar , Diabetes Mellitus Tipo 1/complicaciones , Nefropatías Diabéticas/genética , Nefropatías Diabéticas/orina , Humanos , Interleucina-18/metabolismo , Leucocitos Mononucleares/metabolismo , Factor 88 de Diferenciación Mieloide/genética , Factor 88 de Diferenciación Mieloide/metabolismo , ARN Mensajero/genética , ARN Mensajero/orina , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo , Adulto JovenRESUMEN
Abstract Objective This article presents a clinical and cytogenomic approach that focuses on the diagnosis of syndromic oral clefts (OCs). Methods The inclusion criteria were individuals with OC presenting four or more minor signs and no major defects (non-syndromic oral clefts [NSOCs]) as well as individuals with OC presenting at least another major defect, regardless of the number of minor signs (syndromic oral clefts [SOCs]). The exclusion criteria included NSOC with less than four minor signs, SOC with known etiology, as well as atypical oral clefts. Results Of 1647 individuals with OC recorded in the Brazilian Database of Craniofacial Anomalies, 100 individuals were selected for chromosome microarray analysis (CMA). Among these, 44 individuals were clinically classified as NSOC and 56 as SOC. CMA was performed for both groups, and abnormal CMA was identified in 9%, all previously classified as SCO. The clinical and CMA data analyses showed a significant predominance of abnormal CMA in individuals classified as SOC (p = 0.0044); prematurity, weight, length, and head circumference at birth were significantly lower in the group with abnormal CMA. Besides, minor signs were significantly higher in this group (p = 0.0090). Conclusion The rigorous selection of cases indicates that the significant variables could help in early recognition of SOC. This study reinforces the importance of applying the CMA technique to establish the diagnosis of SOC. This is an important and universal issue in clinical practice for intervention, care, and genetic counseling.
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Humanos , Labio Leporino/genética , Fisura del Paladar/genética , Brasil , Aberraciones Cromosómicas , GenómicaRESUMEN
OBJECTIVE: This article presents a clinical and cytogenomic approach that focuses on the diagnosis of syndromic oral clefts (OCs). METHODS: The inclusion criteria were individuals with OC presenting four or more minor signs and no major defects (non-syndromic oral clefts [NSOCs]) as well as individuals with OC presenting at least another major defect, regardless of the number of minor signs (syndromic oral clefts [SOCs]). The exclusion criteria included NSOC with less than four minor signs, SOC with known etiology, as well as atypical oral clefts. RESULTS: Of 1647 individuals with OC recorded in the Brazilian Database of Craniofacial Anomalies, 100 individuals were selected for chromosome microarray analysis (CMA). Among these, 44 individuals were clinically classified as NSOC and 56 as SOC. CMA was performed for both groups, and abnormal CMA was identified in 9%, all previously classified as SCO. The clinical and CMA data analyses showed a significant predominance of abnormal CMA in individuals classified as SOC (pâ¯=â¯0.0044); prematurity, weight, length, and head circumference at birth were significantly lower in the group with abnormal CMA. Besides, minor signs were significantly higher in this group (pâ¯=â¯0.0090). CONCLUSION: The rigorous selection of cases indicates that the significant variables could help in early recognition of SOC. This study reinforces the importance of applying the CMA technique to establish the diagnosis of SOC. This is an important and universal issue in clinical practice for intervention, care, and genetic counseling.
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Labio Leporino , Fisura del Paladar , Brasil , Aberraciones Cromosómicas , Labio Leporino/genética , Fisura del Paladar/genética , Genómica , Humanos , Recién NacidoRESUMEN
AIM: To evaluate glycaemic control and its influence on albuminuria and bone mineral density (BMD) in children and adolescents with type 1 diabetes (T1D). METHODS: We collectively assessed 84 T1D children/adolescents (T1D group), aged between 6 and 17 years, and then divided them into two groups according to their glycaemic profile (T1D with good glycaemic control (T1DG group) and T1D with poor glycaemic control (T1DP group)). Serum glucose, glycated haemoglobin, serum urea, serum creatinine, urinary albumin-to-creatinine ratio (ACR), estimated glomerular filtration rate and BMD levels were assessed. RESULTS: Of the patients studied, 77% presented with poor glycaemic control. Patients with T1DP showed an increased ACR (P < 0.001) and a low BMD (P = 0.025) when compared to the T1DG group. In addition, five patients in the T1DP group presented with concomitant albuminuria and a low BMD for their chronological age. Significant negative correlations were identified between the ACR and glycated haemoglobin (r = 0.655, P < 0.001), BMD and glycated haemoglobin (r = -0.262, P = 0.047) and BMD and the ACR (r = -0.631, P = <0.001). In linear regression analysis, the ACR showed a negative effect on BMD (P = 0.044) in the T1D patient group. CONCLUSION: Poor glycaemic control was correlated with albuminuria, suggestive of a negative effect on bone tissue, leading to a low BMD in children and adolescents with T1D.
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Albuminuria/epidemiología , Enfermedades Óseas Metabólicas/epidemiología , Diabetes Mellitus Tipo 1/diagnóstico , Diabetes Mellitus Tipo 1/epidemiología , Hemoglobina Glucada/análisis , Índice Glucémico , Adolescente , Factores de Edad , Albuminuria/diagnóstico , Glucemia/análisis , Enfermedades Óseas Metabólicas/diagnóstico por imagen , Brasil , Niño , Bases de Datos Factuales , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Femenino , Tasa de Filtración Glomerular , Hospitales Pediátricos , Humanos , Incidencia , Modelos Lineales , Masculino , Pronóstico , Estudios Retrospectivos , Índice de Severidad de la Enfermedad , Factores Sexuales , Estadísticas no ParamétricasRESUMEN
OBJECTIVES: The present study aims to describe the clinical, electrocardiographic, and echocardiographic cardiological findings in a group of patients with oral clefts. METHODS: This is a prospective cross-sectional study on 70 children (age range from 13 days to 19 years) with oral clefts who attended the multidisciplinary program of a university hospital from March 2013 to September 2014. The patients were evaluated by a pediatric cardiologist and underwent detailed anamnesis, physical examination, electrocardiogram, and echocardiogram. RESULTS: Sixty percent of the patients were male; 55.7% presented with cleft lip and palate, and 40.0% presented with health complaints. Comorbidities were found in 44.3%. Relevant pregnancy, neonatal, family and personal antecedents were present in 55.7%, 27.1%, 67.2%, and 24.3% of the patients, respectively. Regarding the antecedents, 15.2% of the patients presented with a cardiac murmur, 49.0% with a familial risk of developing plurimetabolic syndrome, and 6% with family antecedents of rheumatic fever. Electrocardiographic evaluation showed one case of atrioventricular block. Echocardiograms were abnormal in 35.7% of the exams, including 5 cases of mitral valve prolapse - one of which was diagnosed with rheumatic heart disease. CONCLUSION: The finding of a family risk of developing plurimetabolic syndrome and a diagnosis of rheumatic heart disease indicates that patients with oral clefts may be more prone to developing acquired heart disease. Thus, our findings highlight the importance of anamnesis and methodological triangulation (clinical-electrocardiographic-echocardiographic) in the investigation of patients with oral clefts and emphasize that cardiological follow-up to evaluate acquired and/or rhythm heart diseases is necessary. This strategy permits comorbidity prevention and individualized planned treatment.
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Anomalías Cardiovasculares/complicaciones , Labio Leporino/complicaciones , Fisura del Paladar/complicaciones , Adolescente , Adulto , Anomalías Cardiovasculares/diagnóstico por imagen , Niño , Preescolar , Estudios Transversales , Ecocardiografía , Electrocardiografía , Salud de la Familia , Femenino , Defectos del Tabique Interventricular/complicaciones , Defectos del Tabique Interventricular/diagnóstico por imagen , Humanos , Lactante , Recién Nacido , Masculino , Síndrome Metabólico/complicaciones , Estudios Prospectivos , Medición de Riesgo , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
BACKGROUND: The World Health Organization has recognized the relevance of databases on craniofacial anomalies since . To date, there is no universal standard instrument/database focused on risk factors, clinical and genetic data collection, and follow-up that enables comparison between different populations and genotype-phenotype correlation. Although studies have shown that specific genes would impact outcomes, knowledge is not sufficient to subsidize cost-effectiveness strategies for diagnosis, surgical decision, and a multi-professional approach toward personalized medicine. METHODS: Based on a clinical genetic approach, a Web-based application named CranFlow-Craniofacial Anomalies: Registration, Flow, and Management has been developed. It prospectively collects clinical and genetic information for the Brazilian Database on Craniofacial Anomalies (syndromic and nonsyndromic orofacial cleft, 22q11.2 deletion syndrome, and other craniofacial related disorders). A comprehensive list of CranFlow's features is provided. RESULTS: We present preliminary results on 1546 cases already recorded and followed, which allows recognizing 10% of diagnosis changes. CONCLUSION: The identification of risk factors, consistent genetic approach associated with clinical data and follow-up result in valuable information to develop and improve personalized treatment and studies on genotype-phenotype correlation. Adoption of CranFlow in different clinical services may support comparison between populations. This application has the potential to contribute to improvements in healthcare, quality of services, clinical and surgical outcomes, and the standard of living of individuals with craniofacial anomalies. Birth Defects Research 110:72-80, 2018. © 2017 Wiley Periodicals, Inc.
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Anomalías Craneofaciales/clasificación , Brasil/epidemiología , Bases de Datos Factuales , Estudios de Asociación Genética , Humanos , Sistema de Registros , Programas InformáticosRESUMEN
OBJECTIVES: The present study aims to describe the clinical, electrocardiographic, and echocardiographic cardiological findings in a group of patients with oral clefts. METHODS: This is a prospective cross-sectional study on 70 children (age range from 13 days to 19 years) with oral clefts who attended the multidisciplinary program of a university hospital from March 2013 to September 2014. The patients were evaluated by a pediatric cardiologist and underwent detailed anamnesis, physical examination, electrocardiogram, and echocardiogram. RESULTS: Sixty percent of the patients were male; 55.7% presented with cleft lip and palate, and 40.0% presented with health complaints. Comorbidities were found in 44.3%. Relevant pregnancy, neonatal, family and personal antecedents were present in 55.7%, 27.1%, 67.2%, and 24.3% of the patients, respectively. Regarding the antecedents, 15.2% of the patients presented with a cardiac murmur, 49.0% with a familial risk of developing plurimetabolic syndrome, and 6% with family antecedents of rheumatic fever. Electrocardiographic evaluation showed one case of atrioventricular block. Echocardiograms were abnormal in 35.7% of the exams, including 5 cases of mitral valve prolapse — one of which was diagnosed with rheumatic heart disease. CONCLUSION: The finding of a family risk of developing plurimetabolic syndrome and a diagnosis of rheumatic heart disease indicates that patients with oral clefts may be more prone to developing acquired heart disease. Thus, our findings highlight the importance of anamnesis and methodological triangulation (clinical-electrocardiographic-echocardiographic) in the investigation of patients with oral clefts and emphasize that cardiological follow-up to evaluate acquired and/or rhythm heart diseases is necessary. This strategy permits comorbidity prevention and individualized planned treatment.
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Humanos , Masculino , Femenino , Recién Nacido , Lactante , Preescolar , Niño , Adolescente , Adulto , Adulto Joven , Labio Leporino/complicaciones , Fisura del Paladar/complicaciones , Anomalías Cardiovasculares/complicaciones , Índice de Severidad de la Enfermedad , Ecocardiografía , Salud de la Familia , Estudios Transversales , Estudios Prospectivos , Medición de Riesgo , Anomalías Cardiovasculares/diagnóstico por imagen , Síndrome Metabólico/complicaciones , Electrocardiografía , Defectos del Tabique Interventricular/complicaciones , Defectos del Tabique Interventricular/diagnóstico por imagenRESUMEN
Type 1 diabetes mellitus (T1DM) and estrogen deficiency are associated with several alterations in bone turnover. Zinc (Zn) is required for growth, development, and overall health. Zinc has been used in complementary therapy against bone loss in several diseases. We hypothesized that Zn supplementation represents a potential therapy against severe bone loss induced by the combined effect of estrogen deficiency and T1DM. We evaluated the protective effect of Zn against bone alterations in a chronic model of these disorders. Female Wistar rats were ramdomized into 3 groups (5 rats each): control, OVX/T1DM (ovariectomized rats with streptozotocin-induced T1DM), and OVX/T1DM+Zn (OVX/T1DM plus daily Zn supplementation). Serum biochemical, bone histomorphometric, and molecular analyses were performed. Histomorphometric parameters were similar between the control and OVX/T1DM+Zn groups, suggesting that Zn prevents bone architecture alterations. In contrast, the OVX/T1DM group showed significantly lower trabecular width and bone area as well as greater trabecular separation than the control. The OVX/T1DM and OVX/T1DM+Zn groups had significantly higher serum alkaline phosphatase activity than the control. The supplemented group had higher levels of serum-ionized calcium and phosphorus than the nonsupplemented group. The RANKL/OPG ratio was similar between the control and OVX/T1DM+Zn groups, whereas it was higher in the OVX/T1DM group. In conclusion, Zn supplementation prevents bone alteration in chronic OVX/T1DM rats, as demonstrated by the reduced RANKL/OPG ratio and preservation of bone architecture. The findings may represent a novel therapeutic approach to preventing OVX/T1DM-induced bone alterations.
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Densidad Ósea/efectos de los fármacos , Diabetes Mellitus Experimental/tratamiento farmacológico , Suplementos Dietéticos , Osteoprotegerina/metabolismo , Ligando RANK/metabolismo , Zinc/administración & dosificación , Fosfatasa Alcalina/sangre , Animales , Glucemia/metabolismo , Huesos/efectos de los fármacos , Calcio/sangre , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Femenino , Osteoprotegerina/genética , Ovariectomía , Fósforo/sangre , Ligando RANK/genética , Ratas , Ratas WistarRESUMEN
An integrative analysis of miRNA and mRNA expression profiles in left ventricle (LV) of diabetes-induced rats was performed to elucidate the role of miRNAs and their mRNAs target in diabetic cardiomyopathy (DCM). mRNA (GSE4745) and miRNA (GSE44179) datasets were downloaded from Gene Expression Omnibus 2R (GEO2R) and differentially expressed mRNAs and miRNAs were selected. Cardiotoxicity-related mRNAs (n=7) were analyzed by Ingenuity Pathway Analyses 6 (IPA) and regulatory miRNAs (n=639) were identified using TargetScan 7.1. web dataset. The integrative analysis was performed between miRNAs differentially expressed in GSE44179 and regulatory TargetScan-detected miRNAs of mRNAs differentially expressed in GSE4745. Pla2g2a and Hk2 mRNAs were up-and-down regulated, respectively, in GSE4745 on days 3 and 42 after diabetes-induction. The Pla2g2a regulatory miRNAs, rno-miR-877, rno-miR-320 and rno-miR-214, were down-regulated, and Hk2 regulatory miRNAs, rno-miR-17, rno-miR-187, rno-miR-34a, rno-miR-322, rno-miR-188, rno-miR-532 and rno-miR-21, were up-regulated in GSE44179 dataset. These results are suggestive that Pla2g2a and Hk2 mRNAs and their regulatory miRNAs play a role in DCM pathogenesis and they may be potential circulating biomarkers to detect early cardiovascular complications in diabetic patients.
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Cardiomiopatías/genética , Diabetes Mellitus/genética , MicroARNs/metabolismo , ARN Mensajero/metabolismo , Animales , Cardiomiopatías/metabolismo , Diabetes Mellitus/metabolismo , Regulación hacia Abajo , MicroARNs/genética , ARN Mensajero/genética , RatasRESUMEN
Type 1 diabetes mellitus (T1DM) and estrogen deficiency are associated with several alterations in bone turnover. Zinc (Zn) is required for growth, development, and overall health. Zinc has been used in complementary therapy against bone loss in several diseases. We hypothesized that Zn supplementation represents a potential therapy against severe bone loss induced by the combined effect of estrogen deficiency and T1DM. We evaluated the protective effect of Zn against bone alterations in a chronic model of these disorders. Female Wistar rats were ramdomized into 3 groups (5 rats each): control, OVX/T1DM (ovariectomized rats with streptozotocin-induced T1DM), and OVX/T1DM+Zn (OVX/T1DM plus daily Zn supplementation). Serum biochemical, bone histomorphometric, and molecular analyses were performed. Histomorphometric parameters were similar between the control and OVX/T1DM+Zn groups, suggesting that Zn prevents bone architecture alterations. In contrast, the OVX/T1DM group showed significantly lower trabecular width and bone area as well as greater trabecular separation than the control...
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Diabetes Mellitus , Ratas , ZincRESUMEN
An integrative analysis of miRNA and mRNA expression profiles in left ventricle (LV) of diabetes-induced rats was performed to elucidate the role of miRNAs and their mRNAs target in diabetic cardiomyopathy (DCM). mRNA (GSE4745) and miRNA (GSE44179) datasets were downloaded from Gene Expression Omnibus 2R (GEO2R) and differentially expressed mRNAs and miRNAs were selected. Cardiotoxicity-related mRNAs (n=7) were analyzed by Ingenuity Pathway Analyses 6 (IPA) and regulatory miRNAs (n=639) were identified using TargetScan 7.1. web dataset. The integrative analysis was performed between miRNAs differentially expressed in GSE44179 and regulatory TargetScan-detected miRNAs of mRNAs differentially expressed in GSE4745. Pla2g2a and Hk2 mRNAs were up-and-down regulated, respectively, in GSE4745 on days 3 and 42 after diabetes-induction. The Pla2g2a regulatory miRNAs, rno-miR-877, rno-miR-320 and rno-miR-214, were down-regulated, and Hk2 regulatory miRNAs, rno-miR-17, rno-miR-187, rno-miR-34a, rno-miR-322, rno-miR-188, rno-miR-532 and rno-miR-21, were up-regulated in GSE44179 dataset. These results are suggestive that Pla2g2a and Hk2 mRNAs and their regulatory miRNAs play a role in DCM pathogenesis and they may be potential circulating biomarkers to detect early cardiovascular complications in diabetic patients.
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Animales , Ratas , Cardiomiopatías/genética , Cardiomiopatías/metabolismo , Diabetes MellitusRESUMEN
BACKGROUND: Pioglitazone is a peroxisome proliferator-activated receptor gamma (PPARγ) activator used in the treatment of type 2 diabetes (DM2) patients and it has been suggested that can induce bone loss. Tumor necrosis factor-α (TNFα) and interleukin-6 (IL-6) mRNA expression in blood leukocytes and the relationship with polymorphisms and bone markers in DM2 treated with pioglitazone were investigated. METHODS: DM2 (n=53) and normoglycemic (NG, n=52) individuals were included. DM2 patients were treated with pioglitazone (45 mg/day/16 weeks). mRNA expression was evaluated by real-time polymerase chain reaction (PCR). TNFA -308G>A and IL6 -174G>C polymorphisms were detected by PCR-RFLP and high resolution melting polymerase chain reaction (HRM-PCR). RESULTS: Pioglitazone reduced bone specific alkaline phosphatase (bALP) and increased TNFα in DM2 group (p<0.001). DM2 or pioglitazone did not influence TNFα and IL-6 expression (p>0.05). TNFA -308A allele was associated with reduced basal TNFα mRNA levels in NG and DM2 and reduced alkaline phosphatase (tALP) after treatment (p<0.05). IL6 -174C allele was associated with decreased oral glucose tolerance test (OGTT)-2 h in DM2 individuals (p<0.05). CONCLUSIONS: TNFA -308G >A polymorphism appear to be involved in regulation of gene expression independently of hyperglycemia and its interaction with pioglitazone may modify tALP, a important bone marker. IL6 -174G>C variant is related with reduced risk of postprandial hyperglycemia but not with mRNA expression or bone markers.
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Diabetes Mellitus Tipo 2/tratamiento farmacológico , Regulación de la Expresión Génica/efectos de los fármacos , Hipoglucemiantes/farmacología , Tiazolidinedionas/farmacología , Adulto , Anciano , Fosfatasa Alcalina/metabolismo , Estudios de Casos y Controles , Femenino , Humanos , Interleucina-6/genética , Leucocitos/metabolismo , Masculino , Persona de Mediana Edad , Pioglitazona , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
BACKGROUND: Pioglitazone is a peroxisome proliferator-activated receptor gamma (PPARã) activator used in the treatment of type 2 diabetes (DM2) patients and it has been suggested that can induce bone loss. Tumor necrosis factor-á (TNFá) and interleukin-6 (IL-6) mRNA expression in blood leukocytes and the relationship with polymorphisms and bone markers in DM2 treated with pioglitazone were investigated.METHODS: DM2 (n=53) and normoglycemic (NG, n=52) individuals were included. DM2 patients were treated with pioglitazone (45 mg/day/16 weeks). mRNA expression was evaluated by real-time polymerase chain reaction (PCR). TNFA -308G>A and IL6 -174G>C polymorphisms were detected by PCR-RFLP and high resolution melting polymerase chain reaction (HRM-PCR).RESULTS: Pioglitazone reduced bone specific alkaline phosphatase (bALP) and increased TNFá in DM2 group (p0.05). TNFA -308A allele was associated with reduced basal TNFá mRNA levels in NG and DM2 and reduced alkaline phosphatase (tALP) after treatment (pA polymorphism appear to be involved in regulation of gene expression independently of hyperglycemia and its interaction with pioglitazone may modify tALP, a important bone marker. IL6 -174G>C variant is related with reduced risk of postprandial hyperglycemia but not with mRNA expression or bone markers.
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BACKGROUND: Restriction fragment length polymorphism (RFLP) is a common molecular assay used for genotyping, and it requires validated quality control procedures to prevent mistyping caused by impaired endonuclease activity. We have evaluated the usefulness of a plasmid-based internal control in RFLP assays. RESULTS: Blood samples were collected from 102 individuals with acute myocardial infarction (AMI) and 108 non-AMI individuals (controls) for DNA extraction and laboratory analyses. The 1196C > T polymorphism in the toll-like receptor 4 (TLR4) gene was amplified by mismatched-polymerase chain reaction (PCR). Amplicons and pBluescript II SK- plasmid were simultaneously digested with endonuclease HincII. Fragments were separated on 2% agarose gels. Plasmid was completely digested using up to 55.2 nmL/L DNA solutions and 1 microL PCR product. Nevertheless, plasmid DNA with 41.4 nM or higher concentrations was incompletely digested in the presence of 7 microL PCR product. In standardized conditions, TLR4 1196C>T variant was accurately genotyped. TLR4 1196T allele frequency was similar between AMI (3.1%) and controls (2.0%, p = 0.948). TLR4 SNP was not associated with AMI in this sample population. In conclusion, the plasmid-based control is a useful approach to prevent mistyping in RFLP assays, and it is validate for genetic association studies such as TLR4 1196C>T.
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ADN/genética , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Receptor Toll-Like 4/genética , Adulto , Alelos , Estudios de Casos y Controles , ADN/aislamiento & purificación , Femenino , Frecuencia de los Genes , Humanos , Masculino , Infarto del Miocardio/sangre , Plásmidos , Estándares de ReferenciaRESUMEN
A eletroforese de proteínas permite qualificar e quantificar as proteínas séricas, auxiliando no diagnóstico e controle das alterações patológicas. Este trabalho objetiva estudar o perfil eletroforético das proteínas séricas das crianças atendidas no Hospitalde Pediatria (HOSPED/UFRN) e auxiliar no diagnóstico de patologias decorrentes de insuficiência/disfunção protéica. Foram coletadas amostras de sangue de 98 crianças atendidas no HOSPED, na faixa etária compreendida de 1 dia a 15 anos, e encaminhadas aoLaboratório de Bioquímica Clínica do DACT/CCS, onde se realizou a eletroforese de proteína em suporte de acetato de celulose, posteriormente quantificadas por Densitometria. As amostras avaliadas apresentaram um perfil eletroforético bem definido quanto à separaçãodas frações protéicas albumina, alfa 1, alfa 2, beta e gama globulina. Observou-se que 51 pacientes (52,04%) enquadaramse no Grupo I - processo inflamatório agudo, 32 pacientes (32,66%) no Grupo II - processo crônico e 11 pacientes do Grupo III (11,22%)apresentaram valores normais no padrão eletroforético analisado. Em outros 4 pacientes, observou-se situações isoladas com níveis diminuídos das frações alfa 1 e alfa 2 globulinas, da fração albumínica, ou aumento da fração beta. Estes resultados mostram um perfil preliminar das principais patologias que acometem a população estudada, sendo estes relevantes para monitoramento, auxílio e confirmação do diagnóstico das mesmas.
The electrophoresis of proteins allows to qualify and quantify seric proteins, helping out the diagnosis and control of pathologic alterations. This work aims to study the electrophoretic profile of serum proteins in children assisted at the Pediatric Hospital (HOSPED/UFRN) and aid in the diagnosis of pathologies origined by proteic insufficiency or disfuncion. It had been collected blood samples from children assisted at HOSPED, within the age range of 0 and 15 years of age, and guided to the Laboratory of Clinical Bioquemistry of DACT/CCS, where the electrophoresis of proteins took place in celulosis acetate support, later quantified by Desintometry.The samples evaluated showed an electrophoretic profile well defined regarding the separation of proteic fractions albumin, alfa 1, alfa 2, beta and gama globulins. Among 98 analysed cases, 51 patients (52.04%) were included in G-I (acute inflamatory process), 32 patients (32.66%) in G-II (cronic process) and 11 patients included in G-III (11.22%) showed normal values in the electrophoretic pattern analysed. In other 4 patients, it was observed isolated situations with low levels of alfa 1 and alfa 2 globulins, albumin fraction, or increasing of beta fraction. These results show a preliminary profile of the main pathologies which assalt the studied population, these are relevant to the follow up and confirmation of the diagnosis.
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Humanos , Masculino , Femenino , Lactante , Preescolar , Niño , Adolescente , Proteínas de Fase Aguda , Análisis Químico de la Sangre , Electroforesis de las Proteínas Sanguíneas , Proteínas Sanguíneas , Malaria , Plasmodium falciparumRESUMEN
PURPOSE: Considering that important scientific advances have been obtained through studies based on experimental Diabetes mellitus, and that tamoxifen action in humans remains unknown, the aim of the present work is to follow the modifications promoted by diabetes and tamoxifen in the electrophoretic profile of plasmatic proteins. METHODS: It was used 27 Wistar female rats (180-250 body weight), randomicaly divided into five groups: C1 (n = 3, received vehicle), C2 (n = 3, no treatment), T (n =5, treated with tamoxifen, 0.3mg/Kg/day), D (n = 8, experimental diabetes by estreptozotocin, 45mg/Kg and DT (n = 8, diabetic treated with tamoxifen). The electrophoresis was accomplished in cellulose acetate. pH 8.6-8.8, TECNOW chamber, and the strains were stained by Ponceau S. The total proteins were determined by the Biuret method (Labtest). Proteinograms were obtained in densitometer BioSystems BTS-235. RESULTS: Albumin decreased progressively in the groups T, D and DT; a1 fraction increased in groups T and DT; a2 fraction increased in groups T and D, including a synergic effect in group DT; a fraction increased in groups T and D; a fraction increased in groups T, D and DT. CONCLUSIONS: The results indicate an acute phase resposta, with synergic effect of tamoxifen and diabetes, suggesting a probable hepatic lesion.
Asunto(s)
Proteínas Sanguíneas/efectos de los fármacos , Diabetes Mellitus Experimental/sangre , Diabetes Mellitus Tipo 1/sangre , Antagonistas de Estrógenos/farmacología , Tamoxifeno/farmacología , Análisis de Varianza , Animales , Glucemia/efectos de los fármacos , Electroforesis de las Proteínas Sanguíneas , Peso Corporal/efectos de los fármacos , Ciclo Estral/efectos de los fármacos , Femenino , Ratas , Ratas WistarRESUMEN
Although osteopenia has been associated with human diabetes mellitus, the pathogenesis of diabetic osteopenia is unclear. In the present study, we evaluated the effect of diabetes on histomorphometry, bone mineral density (BMD)-measured by dual-energy X-ray absorptiometry (DXA)-and biomarkers of bone metabolism in rats up to 120 days after the onset of experimental diabetes. Female Wistar rats with a regular estrous cycle were randomly divided into two groups: control rats (n = 15) and diabetic rats without insulin treatment (n = 25). Diabetes was induced by injection of alloxan and was confirmed by the determination of blood glucose concentration (>250 mg/dl). The results revealed an approximate threefold increase of femoral trabecular distance in diabetic rats compared to controls. Conversely, trabecular thickness and bone trabecular volume were reduced twofold and 77%, respectively. BMD in both the metadiaphyseal region and total area of the femur was found to be clearly reduced in diabetic animals, with no significant differences between the groups. Serum alkaline phosphatase (ALP) and tartarate-resistant acid phosphatase (TRAP) activities showed significant six- and twofold increases, respectively, in diabetic rats. There were significant decreases in serum calcium and albumin concentrations in diabetic rats, but no difference was observed in serum magnesium, phosphorus, or creatinine concentrations between the groups. Overall, our findings support the conclusion that the diabetic state is associated with alterations in bone turnover, resulting in the development of osteopenia, which is related to the time of evolution of the disorder.
Asunto(s)
Enfermedades Óseas Metabólicas/etiología , Enfermedades Óseas Metabólicas/patología , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/patología , Fosfatasa Ácida/sangre , Fosfatasa Alcalina/sangre , Animales , Biomarcadores/análisis , Biomarcadores/sangre , Glucemia/metabolismo , Densidad Ósea/fisiología , Enfermedades Óseas Metabólicas/metabolismo , Huesos/metabolismo , Huesos/patología , Calcio/sangre , Creatina/sangre , Diabetes Mellitus Experimental/sangre , Diabetes Mellitus Experimental/inducido químicamente , Ciclo Estral , Femenino , Isoenzimas/sangre , Ratas , Ratas Wistar , Albúmina Sérica/análisis , Fosfatasa Ácida TartratorresistenteRESUMEN
OBJETIVO: Considerando-se que importantes avanços científicos têm sido obtidos através de estudos com Diabetes mellitus experimental, e que a ação do tamoxifeno em humanos permanece obscura, o presente trabalho objetiva acompanhar as modificações promovidas pelo diabetes e tamoxifeno no perfil eletroforético das proteínas plasmáticas. MÉTODOS: Foram utilizados 27 ratos fêmeas Wistar (180-220g peso corporal), divididos randomicamente em 5 grupos: C1 (n=3, receberam veículo), C2 (n=3, sem tratamento), T (n=5, tratados com tamoxifeno, 0,3mg/kg/dia), D (n=8, diabéticos experimentais por estreptozotocina, 45mg/Kg) e DT (n=8, diabéticos tratados com tamoxifeno). A eletroforese foi realizada em acetato de celulose, pH 8,6-8,8, cuba TECNOW, e as fitas foram coradas em Ponceau S. As proteínas totais foram determinadas pelo método do Biureto (Kit Labtest). Os proteinogramas foram obtidos em densitômetro BioSystems BTS-235. RESULTADOS: Albumina diminuiu progressivamente nos grupos T, D e DT; a fração a1 aumentou nos grupos T e DT; a fração a2 aumentou nos grupos T e D, havendo efeito aditivo no grupo DT; a fração b aumentou nos grupos T e D; a fração g aumentou nos grupos T, D e DT. CONCLUSÃO: Os resultados indicam uma resposta de fase aguda, com efeito aditivo do tamoxifeno e diabetes, sugerindo uma provável lesão hepática.
Asunto(s)
Animales , Femenino , Ratas , Proteínas Sanguíneas/efectos de los fármacos , Diabetes Mellitus Experimental/sangre , Diabetes Mellitus Tipo 1/sangre , Antagonistas de Estrógenos/farmacología , Tamoxifeno/farmacología , Análisis de Varianza , Electroforesis de las Proteínas Sanguíneas , Glucemia/efectos de los fármacos , Peso Corporal/efectos de los fármacos , Ciclo Estral/efectos de los fármacos , Ratas WistarRESUMEN
We report the kinetic characterization of an ecto-nucleosidetriphosphate diphosphohydrolase 1 from rat osseous plate membranes in streptozotocin-induced diabetic rats, which arises during ectopic mineralization twenty days after a subcutaneous implantation of demineralized bone matrix, Insulin deficiency decreased the ecto-nucleoside triphosphate diphosphohydrolase activity from 1293.1 +/- 39.8 (control rats) to 556.0 +/- 8.2 nmol Pi/(min mg). Two families of ATP hydrolyzing sites showed cooperative effects with specific activities of 256.2 +/- 7.7 nmol Pi/(min mg) and 299.8 +/- 8.9 nmol Pi/(min mg), and studies on the stimulation of the enzyme by magnesium and calcium ions showed that the decrease in enzyme activity results from changes in the affinity of the enzyme for these ions. To our knowledge this is the first study associating the effects of type I diabetes with an ecto-nucleoside triphosphate diphosphohydrolase activity from rat osseous plate membranes.