Gene therapy for diabetes mellitus in rats by hepatic expression of insulin.

Type 1 diabetes mellitus is caused by severe insulin deficiency secondary to the autoimmune destruction of pancreatic beta cells. Patients need to be controlled by periodic insulin injections to prevent the development of ketoacidosis, which can be fatal. Sustained, low-level expression of the rat insulin 1 gene from the liver of severely diabetic rats was achieved by in vivo administration of a recombinant retroviral vector. Ketoacidosis was prevented and the treated animals exhibited normoglycemia during a 24-hr fast, with no evidence of hypoglycemia. Histopathological examination of the liver in the treated animals showed no apparent abnormalities. Thus, the liver is an excellent target organ for ectopic expression of the insulin gene as a potential treatment modality for type 1 diabetes mellitus by gene therapy.

Efeito dos ácidos graxos ômega-3, ômega-6 e ômega-9 sobre o risco cardiovascular de indivíduos adultos: estudo clínico de prevenção primária.

INTRODUÇÃO: As doenças cardiovasculares (DCV) são a principal causa de morte no mundo, sendo muitos dos fatores de risco passíveis de prevenção e controle. Embora as DCV sejam complexas em sua etiologia e desenvolvimento, a concentração elevada de LDL-c e baixa de HDL-c constituem os fatores de risco modificáveis mais monitorados na prática clínica, embora não sejam capazes de explicar todos os eventos cardiovasculares. Portanto, investigar como intervenções farmacológicas e nutricionais podem modular parâmetros oxidativos, físicos e estruturais das lipoproteínas pode fornecer estimativa adicional ao risco cardiovascular. Dentre os diversos nutrientes e compostos bioativos relacionados às DCV, os lipídeos representam os mais investigados e descritos na literatura. Nesse contexto, os ácidos graxos insaturados (ômega-3, ômega-6 e ômega-9) têm sido foco de inúmeros estudos. OBJETIVOS: Avaliar o efeito da suplementação com ômega-3, ômega-6 e ômega-9 sobre os parâmetros cardiometabólicos em indivíduos adultos com múltiplos fatores de risco e sem evento cardiovascular prévio. MATERIAL E MÉTODOS: Estudo clínico, randomizado, duplo-cego, baseado em intervenção nutricional (3,0 g/dia de ácidos graxos) sob a fórmula de cápsulas contendo: ômega-3 (37 por cento de EPA e 23 por cento de DHA) ou ômega-6 (65 por cento de ácido linoleico) ou ômega-9 (72 por cento de ácido oleico). A amostra foi composta por indivíduos de ambos os sexos, com idade entre 30 e 74 anos, apresentando pelo menos um dos seguintes fatores de risco: Dislipidemia, Diabetes Mellitus, Obesidade e Hipertensão Arterial Sistêmica. Após aprovação do Comitê de Ética, os indivíduos foram distribuídos nos três grupos de intervenção. No momento basal, os indivíduos foram caracterizados quanto aos aspectos demográficos (sexo, idade e etnia) e clínicos (medicamentos, doenças atuais e antecedentes familiares). Nos momentos basal e após 8 semanas de intervenção, amostras de sangue foram coletadas após 12h de jejum. A partir do plasma foram analisados: perfil lipídico (CT, LDL-c, HDL-c, TG), apolipoproteínas AI e B, ácidos graxos não esterificados, atividade da PON1, LDL(-) e auto-anticorpos, ácidos graxos, glicose, insulina, tamanho e distribuição percentual da LDL (7 subfrações e fenótipo A e não-A) e HDL (10 subfrações). O efeito do tempo, da intervenção e associações entre os ácidos graxos e aspectos qualitativos das lipoproteínas foram testados (SPSS versão 20.0, p <0,05). RESULTADOS: Uma primeira análise dos resultados baseada em um corte transversal demonstrou, por meio da análise de tendência linear ajustada pelo nível de risco cardiovascular, que o maior tercil plasmático de DHA se associou positivamente com HDL-c, HDLGRANDE e tamanho de LDL e negativamente com HDLPEQUENA e TG. Observou-se também que o maior tercil plasmático de ácido linoleico se associou positivamente com HDLGRANDE e tamanho de LDL e negativamente com HDLPEQUENA e TG. Esse perfil de associação não foi observado quando foram avaliados os parâmetros dietéticos. Avaliando uma subamostra que incluiu indivíduos tabagistas suplementados com ômega-6 e ômega-3, observou-se que ômega-3 modificou positivamente o perfil lipídico e as subfrações da HDL. Nos modelos de regressão linear ajustados pela idade, sexo e hipertensão, o DHA plasmático apresentou associações negativas com a HDLPEQUENA. Quando se avaliou exclusivamente o efeito do ômega-3 em indivíduos tabagistas e não tabagistas, observou-se que fumantes, do sexo masculino, acima de 60 anos de idade, apresentando baixo percentual plasmático de EPA e DHA (<8 por cento ), com excesso de peso e gordura corporal elevada, apresentam maior probabilidade de ter um perfil de subfrações de HDL mais aterogênicas. Tendo por base os resultados acima, foi comparado o efeito do ômega-3, ômega-6 e ômega-9 sobre os parâmetros cardiometabólicos. O ômega-3 promoveu redução no TG, aumento do percentual de HDLGRANDE e redução de HDLPEQUENA. O papel cardioprotetor do ômega-3 foi reforçado pelo aumento na incorporação de EPA e DHA, no qual indivíduos com EPA e DHA acima de 8 por cento apresentaram maior probabilidade de ter HDLGRANDE e menor de ter HDLPEQUENA. Em adição, observou-se também que o elevado percentual plasmático de ômega-9 se associou com partículas de LDL menos aterogênicas (fenótipo A). CONCLUSÃO: Ácidos graxos plasmáticos, mas não dietéticos, se correlacionam com parâmetros cardiometabólicos. A suplementação com ômega-3, presente no óleo de peixe, promoveu redução no TG e melhoria nos parâmetros qualitativos da HDL (mais HDLGRANDE e menos HDLPEQUENA). Os benefícios do ômega-3 foram particularmente relevantes nos indivíduos tabagistas e naqueles com menor conteúdo basal de EPA e DHA plasmáticos. Observou-se ainda que o ômega-9 plasmático, presente no azeite de oliva, exerceu impacto positivo no tamanho e subfrações da LDL.

Insulin; its use in the treatment of diabetes.

Physiology, by J. J. R. Macleod.--Clinical section, by W. R. Campbell.

Increased C282Y heterozygosity in gestational diabetes

Background. Hereditary hemochromatosis is an autosomal recessive disorder of iron metabolism that is characterized by excess accumulation of iron in various organs and often leads to diabetes mellitus (DM). To study whether mutations in the hemochromatosis gene (HFE) could be a risk factor for the development of gestational diabetes mellitus (GDM), the prevalence of HFE mutations in patients with GDM was compared to that of healthy pregnant controls. Methods: GDM was diagnosed in 208 of 2,421 pregnant woman screened between the 24th and 28th week of gestation over a period of 18 months. Patients and 170 matched control subjects were screened for the HFE gene mutations C282Y and H63D. Results: In North and Central European GDM patients, the allele frequency of the C282Y mutation (7.7%) was higher than in pregnant controls (2.9%; p = 0.04), while the frequency of the H63D mutation was not different (p = 0.45). Three patients with GDM were homozygous for H63D (3.1%), 1 patient was homozygous for C282Y (1.0%), 2 patients were compound heterozygous (2.0%) and 26 were heterozygous [11 C282Y (11.2%) and 15 H63D (15.3%)]. C282Y and H63D allele frequencies were not different between controls and GDIVI patients of Southern European or non-European origin. Irrespective of the HIFE-mutation status, serum ferritin levels were increased in patients with GDM compared to healthy pregnant controls (p = 0.01), while transferrin saturation was similar in both groups. Conclusions: In North and Central European patients with GDM, the C282Y allele frequency is higherthan in healthy pregnant women, suggesting a genetic susceptibility to the development of GDM. Copyright (c) 2005 S. Karger AG, Basel.

Evidence for a non-antioxidant, dose-dependent role of alpha-lipoic acid in caspase-3 and ERK2 activation in endothelial cells

Endothelial cell apoptosis contributes to atherosclerosis and may be exacerbated by oxidative stress. Results from clinical trials using antioxidant supplementation are equivocal and could be enhanced by antioxidants with additional non-antioxidant properties such as a-lipoic acid and alpha-tocopherol. The aim of this study was to investigate the effects of these antioxidants on cytoprotective pathways and endothelial apoptosis. Endothelial cells were incubated with alpha-lipoic acid and alpha-tocopherol, alone or in combination, prior to incubation with H2O2 or staurosporine. alpha-lipoic acid pre-treatment alone increased caspase-3 activity in a dose-dependent manner. Both H2O2 and staurosporine increased DNA fragmentation and caspase-3 activity and pre-treatment of cells with a-lipoic acid and/or a-tocopherol failed to prevent stress-induced apoptosis. Neither antioxidant treatments nor apoptotic inducers alone altered expressions of BcI-2, Bax, HSP70 or pERK1/2 or pJNK. alpha-lipoic decreased pERK2 in staurosporine-treated cells in a dose-dependent manner. These findings indicate that pre-incubation with alpha-lipoic acid and alpha-tocopherol, alone or in combination, does not protect against oxidative- or non-oxidative-induced apoptosis in endothelial cells. Moreover, we have demonstrated a non-antioxidant, dose-dependent role of alpha-lipoic acid in caspase-3 and ERK2 activation. These data provide an insight and indicate caution in the use of high doses of alpha-lipoic acid as an antioxidant.

Carotid artery intimal medial thickness, brachial artery flow-mediated vasodilation and cardiovascular risk factors in diabetic and non-diabetic indigenous Australians

Background: Indigenous Australians are at high risk for cardiovascular disease and type 2 diabetes. Carotid artery intimal medial thickness (CIMT) and brachial artery flow-mediated vasodilation (FMD) are ultrasound imaging based surrogate markers of cardiovascular risk. This study examines the relative contributions of traditional cardiovascular risk factors on CIMT and FMD in adult Indigenous Australians with and without type 2 diabetes mellitus. Method: One hundred and nineteen Indigenous Australians were recruited. Physical and biochemical markers of cardiovascular risk, together with CIMT and FMD were meausred for all subjects. Results: Fifty-three Indigenous Australians subjects (45%) had type 2 diabetes mellitus. There was a significantly greater mean CIMT in diabetic versus non-diabetic subjects (p = 0.049). In the non-diabetic group with non-parametric analyses, there were significant correlations between CIMT and: age (r = 0.64, p < 0.001), systolic blood pressure (r = 0.47, p < 0.001) and non-smokers (r = -0.30, p = 0.018). In the diabetic group, non-parametric analysis showed correlations between CIMT, age (r = 0.36, p = 0.009) and duration of diabetes (r = 0.30, p = 0.035) only. Adjusting forage, sex, smoking and history of cardiovascular disease, Hb(A1c) became the sole significant correlate of CIMT (r = 0.35,p = 0.01) in the diabetic group. In non-parametric analysis, age was the sole significant correlate of FMD (r = -0.31,p = 0.013), and only in non-diabetic subjects. Linear regression analysis showed significant associations between CIMT and age (t = 4.6,p < 0.001), systolic blood pressure (t = 2.6, p = 0.010) and Hb(A1c) (t = 2.6, p = 0.012), smoking (t = 2.1, p = 0.04) and fasting LDL-cholesterol (t = 2.1, p = 0.04). There were no significant associations between FMD and examined cardiovascular risk factors with linear regression analysis Conclusions: CIMT appears to be a useful surrogate marker of cardiovascular risk in this sample of Indigenous Australian subjects, correlating better than FMD with established cardiovascular risk factors. A lifestyle intervention programme may alleviate the burden of cardiovascular disease in Indigenous Australians by reducing central obesity, lowering blood pressure, correcting dyslipidaemia and improving glycaemic control. CIMT may prove to be a useful tool to assess efficacy of such an intervention programme. (c) 2004 Elsevier Ireland Ltd. All rights reserved.

Psychosocial and non-psychosocial risk factors for the new diagnosis of diabetes in elderly women

Objectives: Determine psychosocial variables associated with the new diagnosis of diabetes in elderly women. Examine whether variables remained significant predictors after controlling for non-psychosocial risk factors and the frequency of doctor visits. Research design and methods: A longitudinal cohort study was conducted using data from 10 300 women who completed a survey in 1996 and 1999. The women were aged between 70 and 74 years of age in 1996. The were asked to provide self-reports on a number of psychosocial and non-psychosocial variables in 1996 and on whether they had been diagnosed for the first time with diabetes in the 3-year period. The relationships between the potential risk factors and new diagnosis of diabetes were examined using binary logistic regression analysis. Results: Univariate results showed that not having a current partner, having low social support and having a mental health index score in the clinical range were all associated with higher risks of being diagnosed with diabetes for the first time. However the multivariate results showed that only a mental health index score in the clinical range and not having a current partner provided unique prediction of being newly diagnosed with diabetes. Of the non-psychosocial variables measured, only having a high BMI and hypertension were associated with increased risks of new diagnosis, while there was also evidence of a U shaped relationship between alcohol consumption and new diagnosis. Even after adjusting for frequency of doctor visits and non-psychosocial risk factors, a mental health index in the clinical range proved to still be a significant risk factor. Conclusions: A score on the mental health index that is within the clinical range is an independent risk factor for the new diagnosis of diabetes in elderly women. (c) 2006 Elsevier Ireland Ltd. All rights reserved.

Adiponectin multimerization is dependent on conserved lysines in the collagenous domain: Evidence for regulation of multimerization by alterations in posttranslational modifications

Adiponectin is a secreted, multimeric protein with insulin-sensitizing, antiatherogenic, and antiinflammatory properties. Serum adiponectin consists of trimer, hexamer, and larger high-molecular-weight (HMW) multimers, and these HMW multimers appear to be the more bioactive forms. Multimer composition of adiponectin appears to be regulated; however, the molecular mechanisms involved are unknown. We hypothesize that regulation of adiponectin multimerization and secretion occurs via changes in posttranslational modifications (PTMs). Although a structural role for intertrimer disulfide bonds in the formation of hexamers and HMW multimers is established, the role of other PTMs is unknown. PTMs identified in murine and bovine adiponectin include hydroxylation of multiple conserved proline and lysine residues and glycosylation of hydroxylysines. By mass spectrometry, we confirmed the presence of these PTMs in human adiponectin and identified three additional hydroxylations on Pro71, Pro76, and Pro95. We also investigated the role of the five modified lysines in multimer formation and secretion of recombinant human adiponectin expressed in mammalian cell lines. Mutation of modified lysines in the collagenous domain prevented formation of HMW multimers, whereas a pharmacological inhibitor of prolyl- and lysyl-hydroxylases, 2,2'-dipyridyl, inhibited formation of hexamers and HMW multimers. Bacterially expressed human adiponectin displayed a complete lack of differentially modified isoforms and failed to form bona fide trimers and larger multimers. Finally, glucose-induced increases in HMW multimer production from human adipose explants correlated with changes in the two-dimensional electrophoresis profile of adiponectin isoforms. Collectively, these data suggest that adiponectin multimer composition is affected by changes in PTM in response to physiological factors.

Does high intensity resistance training maintain bone mass during moderate weight loss in older overweight adults with type 2 diabetes

The aim was to investigate whether the addition of supervised high intensity progressive resistance training to a moderate weight loss program (RT+WLoss) could maintain bone mineral density (BMD) and lean mass compared to moderate weight loss (WLoss) alone in older overweight adults with type 2 diabetes. We also investigated whether any benefits derived from a supervised RT program could be sustained through an additional home-based program. This was a 12-month trial in which 36 sedentary, overweight adults aged 60 to 80 years with type 2 diabetes were randomized to either a supervised gymnasium-based RT+WLoss or WLoss program for 6 months (phase 1). Thereafter, all participants completed an additional 6-month home-based training without further dietary modification (phase 2). Total body and regional BMD and bone mineral content (BMC), fat mass (FM) and lean mass (LM) were assessed by DXA every 6 months. Diet, muscle strength (1-RM) and serum total testosterone, estradiol, SHBG, insulin and IGF-1 were measured every 3 months. No between group differences were detected for changes in any of the hormonal parameters at any measurement point. In phase 1, after 6 months of gymnasium-based training, weight and FM decreased similarly in both groups (P < 0.01), but LM tended to increase in the RT+WLoss (n=16) relative to the WLoss (n = 13) group [net difference (95% CI), 1.8% (0.2, 3.5), P < 0.05]. Total body BMD and BMC remained unchanged in the RT+WLoss group, but decreased by 0.9 and 1.5%, respectively, in the WLoss group (interaction, P < 0.05). Similar, though non-significant, changes were detected at the femoral neck and lumbar spine (L2-L4). In phase 2, after a further 6 months of home-based training, weight and FM increased significantly in both the RT+WLoss (n = 14) and WLoss (n = 12) group, but there were no significant changes in LM or total body or regional BMD or BMC in either group from 6 to 12 months. These results indicate that in older, overweight adults with type 2 diabetes, dietary modification should be combined with progressive resistance training to optimize the effects on body composition without having a negative effect on bone health.

Diagnosis of diabetes mellitus: case of different cutoff values: A data mining approach

Objective: An estimation of cut-off points for the diagnosis of diabetes mellitus (DM) based on individual risk factors. Methods: A subset of the 1991 Oman National Diabetes Survey is used, including all patients with a 2h post glucose load >= 200 mg/dl (278 subjects) and a control group of 286 subjects. All subjects previously diagnosed as diabetic and all subjects with missing data values were excluded. The data set was analyzed by use of the SPSS Clementine data mining system. Decision Tree Learners (C5 and CART) and a method for mining association rules (the GRI algorithm) are used. The fasting plasma glucose (FPG), age, sex, family history of diabetes and body mass index (BMI) are input risk factors (independent variables), while diabetes onset (the 2h post glucose load >= 200 mg/dl) is the output (dependent variable). All three techniques used were tested by use of crossvalidation (89.8%). Results: Rules produced for diabetes diagnosis are: A- GRI algorithm (1) FPG>=108.9 mg/dl, (2) FPG>=107.1 and age>39.5 years. B- CART decision trees: FPG >=110.7 mg/dl. C- The C5 decision tree learner: (1) FPG>=95.5 and 54, (2) FPG>=106 and 25.2 kg/m2. (3) FPG>=106 and =133 mg/dl. The three techniques produced rules which cover a significant number of cases (82%), with confidence between 74 and 100%. Conclusion: Our approach supports the suggestion that the present cut-off value of fasting plasma glucose (126 mg/dl) for the diagnosis of diabetes mellitus needs revision, and the individual risk factors such as age and BMI should be considered in defining the new cut-off value.