Alterations of specific biomarkers of metabolic pathways in vascular tree from patients with Type 2 diabetes.

The aims of this study were to check whether different biomarkers of inflammatory, apoptotic, immunological or lipid pathways had altered their expression in the occluded popliteal artery (OPA) compared with the internal mammary artery (IMA) and femoral vein (FV) and to examine whether glycemic control influenced the expression of these genes. The study included 20 patients with advanced atherosclerosis and type 2 diabetes mellitus, 15 of whom had peripheral arterial occlusive disease (PAOD), from whom samples of OPA and FV were collected. PAOD patients were classified based on their HbA1c as well (HbA1c ≤ 6.5) or poorly (HbA1c > 6.5) controlled patients. Controls for arteries without atherosclerosis comprised 5 IMA from patients with ischemic cardiomyopathy (ICM). mRNA, protein expression and histological studies were analyzed in IMA, OPA and FV. After analyzing 46 genes, OPA showed higher expression levels than IMA or FV for genes involved in thrombosis (F3), apoptosis (MMP2, MMP9, TIMP1 and TIM3), lipid metabolism (LRP1 and NDUFA), immune response (TLR2) and monocytes adhesion (CD83). Remarkably, MMP-9 expression was lower in OPA from well-controlled patients. In FV from diabetic patients with HbA1c ≤6.5, gene expression levels of BCL2, CDKN1A, COX2, NDUFA and SREBP2 were higher than in FV from those with HbA1c >6.5. The atherosclerotic process in OPA from diabetic patients was associated with high expression levels of inflammatory, lipid metabolism and apoptotic biomarkers. The degree of glycemic control was associated with gene expression markers of apoptosis, lipid metabolism and antioxidants in FV. However, the effect of glycemic control on pro-atherosclerotic gene expression was very low in arteries with established atherosclerosis.

Gut microbiota in children with type 1 diabetes differs from that in healthy children: a case-control study.

BACKGROUND A recent study using a rat model found significant differences at the time of diabetes onset in the bacterial communities responsible for type 1 diabetes modulation. We hypothesized that type 1 diabetes in humans could also be linked to a specific gut microbiota. Our aim was to quantify and evaluate the difference in the composition of gut microbiota between children with type 1 diabetes and healthy children and to determine the possible relationship of the gut microbiota of children with type 1 diabetes with the glycemic level. METHODS A case-control study was carried out with 16 children with type 1 diabetes and 16 healthy children. The fecal bacteria composition was investigated by polymerase chain reaction-denaturing gradient gel electrophoresis and real-time quantitative polymerase chain reaction. RESULTS The mean similarity index was 47.39% for the healthy children and 37.56% for the children with diabetes, whereas the intergroup similarity index was 26.69%. In the children with diabetes, the bacterial number of Actinobacteria and Firmicutes, and the Firmicutes to Bacteroidetes ratio were all significantly decreased, with the quantity of Bacteroidetes significantly increased with respect to healthy children. At the genus level, we found a significant increase in the number of Clostridium, Bacteroides and Veillonella and a significant decrease in the number of Lactobacillus, Bifidobacterium, Blautia coccoides/Eubacterium rectale group and Prevotella in the children with diabetes. We also found that the number of Bifidobacterium and Lactobacillus, and the Firmicutes to Bacteroidetes ratio correlated negatively and significantly with the plasma glucose level while the quantity of Clostridium correlated positively and significantly with the plasma glucose level in the diabetes group. CONCLUSIONS This is the first study showing that type 1 diabetes is associated with compositional changes in gut microbiota. The significant differences in the number of Bifidobacterium, Lactobacillus and Clostridium and in the Firmicutes to Bacteroidetes ratio observed between the two groups could be related to the glycemic level in the group with diabetes. Moreover, the quantity of bacteria essential to maintain gut integrity was significantly lower in the children with diabetes than the healthy children. These findings could be useful for developing strategies to control the development of type 1 diabetes by modifying the gut microbiota.

Telemedicine influence on the follow-up of type 2 diabetes patients.

OBJECTIVE This study was designed to evaluate the impact of a teleassistance system on the metabolic control of type 2 diabetes patients. RESEARCH DESIGN AND METHODS We conducted a 1-year controlled parallel-group trial comparing patients randomized (1) to an intervention group, assigned to a teleassistance system using real-time transmission of blood glucose results, with immediate reply when necessary, and telephone consultations, or (2) to a control group, being regularly followed-up at their healthcare center. Study subjects were type 2 diabetes patients >30 years of age followed in the primary care setting. RESULTS A total of 328 type 2 diabetes patients were recruited from 35 family practices in the province of Málaga, Spain. There was a reduction in hemoglobin A1c after 12 months from 7.62 +/- 1.60% to 7.40 +/- 1.43% (P = 0.027) in the intervention group and from 7.44 +/- 1.31% to 7.35 +/- 1.38% (P = 0.303) in the control group. The difference in the change between groups was not statistically significant. There was also a significant decrease in systolic and diastolic blood pressure, total cholesterol, low-density lipoprotein cholesterol, and body mass index in the intervention group. In the control group, the only significant decline was in low-density lipoprotein cholesterol. CONCLUSIONS A teleassistance system using real-time transmission of blood glucose results with an option to make telephone consultations is feasible in the primary care setting as a support tool for family physicians in their follow-up of type 2 diabetes patients.

Association of HbA1c and cardiovascular and renal disease in an adult Mediterranean population

BACKGROUND: Increasing evidence suggests a mechanistic link between the glycemic environment and renal and cardiovascular events, even below the threshold for diabetes. We aimed to assess the association between HbA1c and chronic kidney disease (CKD) and cardiovascular disease (CVD). METHODS: A cross-sectional study involving a random representative sample of 2270 adults from southern Spain (Malaga) was undertaken. We measured HbA1c, serum creatinine and albuminuria in fasting blood and urine samples. RESULTS: Individuals without diabetes in the upper HbA1c tertile had an unfavorable cardiovascular and renal profile and shared certain clinical characteristics with the patients with diabetes. Overall, a higher HbA1c concentration was strongly associated with CKD or CVD after adjustment for traditional risk factors. The patients with known diabetes had a 2-fold higher odds of CKD or CVD. However, when both parameters were introduced in the same model, the HbA1c concentration was only significantly associated with clinical endpoints (OR: 1.4, 95% CI, 1.1-1.6, P = 0.002). An increase in HbA1c of one percentage point was associated with a 30% to 40% increase in the rate of CKD or CVD. This relationship was apparent in persons with and without known diabetes. ROC curves illustrated that a HbA1c of 37 mmol/mol (5.5%) was the optimal value in terms of sensitivity and specificity for predicting endpoints in this population. CONCLUSION: HbA1c levels were associated with a higher prevalence of CKD and CVD cross-sectionally, regardless of diabetes status. These data support the value of HbA1c as a marker of cardiovascular and renal disease in the general population.

Peripheral arterial disease, type 2 diabetes and postprandial lipidaemia: Is there a link?

Peripheral arterial disease, manifested as intermittent claudication or critical ischaemia, or identified by an ankle/brachial index < 0.9, is present in at least one in every four patients with type 2 diabetes mellitus. Several reasons exist for peripheral arterial disease in diabetes. In addition to hyperglycaemia, smoking and hypertension, the dyslipidaemia that accompanies type 2 diabetes and is characterised by increased triglyceride levels and reduced high-density lipoprotein cholesterol concentrations also seems to contribute to this association. Recent years have witnessed an increased interest in postprandial lipidaemia, as a result of various prospective studies showing that non-fasting triglycerides predict the onset of arteriosclerotic cardiovascular disease better than fasting measurements do. Additionally, the use of certain specific postprandial particle markers, such as apolipoprotein B-48, makes it easier and more simple to approach the postprandial phenomenon. Despite this, only a few studies have evaluated the role of postprandial triglycerides in the development of peripheral arterial disease and type 2 diabetes. The purpose of this review is to examine the epidemiology and risk factors of peripheral arterial disease in type 2 diabetes, focusing on the role of postprandial triglycerides and particles.

Comparación de tres métodos de medición de hemoglobina en cirugía cardiaca

Durante la cirugía cardiaca con circulación extracorpórea (CEC) el umbral transfusional hemático está basado en el valor de las cifras de hemoglobina y/o hematocrito. Estos valores se obtienen mediante las máquinas tipo “point-of-care testing” (POCT) que están presentes en quirófano y en las unidades de reanimación. En nuestro centro hay distintos tipos de máquinas POCT. Todas miden la cantidad de hemoglobina y/o el porcentaje de hematocrito pero cada una de ellas utiliza una metodología diferente para medir los parámetros sanguíneos: (i) la conductividad, y (ii) la espectrofotometría. En este trabajo comparamos cada uno de los POCT con respecto a la máquina de referencia durante la cirugía cardiaca con CEC, buscando (1) la posible existencia de errores sistemáticos en los POCT al medir las cifras de hemoglobina y hematocrito, y (2) la validez de cada POCT para detector el umbral de transfusión.

Detection of possible variant forms of hemoglobin in HbA1c measurements by chromatography liquid ion exchange high resolution (HPLC)

Background: The glycosylated hemoglobin (HbA1c) is used to help monitor the degree of a diabetic’s hyperglycemia. Security and accuracy of the methods used in its detection are affected by variants forms of Hb or elevations in levels of Fetal Hb (HbF). These interference are the result of a change in the haemoglobin total net charge of the variant due of a substitution of one amino acid in the remaining amino terminal of the beta chain. International Standardization for HbA1c values (NGSP) not include interference assessment as part of the certification program. Therefore, the effect of each variant or the lifting of the HbF on HbA1c result should be examined in each sample depending on the detected variant and the method used for the detection of the same. The objectives were: to describe the possible variants of Hb and their interference in HbA1c measurement by our method, after the implementation of a computer program for their detection. To identify some variants detected by chromatography liquid ion exchange high resolution (HPLC) with DNA molecular sequencing.

Evaluation of two HbA1c point-of-care analyzers.

BACKGROUND Measurement of HbA1c is the most important parameter to assess glycemic control in diabetic patients. Different point-of-care devices for HbA1c are available. The aim of this study was to evaluate two point-of-care testing (POCT) analyzers (DCA Vantage from Siemens and Afinion from Axis-Shield). We studied the bias and precision as well as interference from carbamylated hemoglobin. METHODS Bias of the POCT analyzers was obtained by measuring 53 blood samples from diabetic patients with a wide range of HbA1c, 4%-14% (20-130 mmol/mol), and comparing the results with those obtained by the laboratory method: HPLC HA 8160 Menarini. Precision was performed by 20 successive determinations of two samples with low 4.2% (22 mmol/mol) and high 9.5% (80 mmol/mol) HbA1c values. The possible interference from carbamylated hemoglobin was studied using 25 samples from patients with chronic renal failure. RESULTS The means of the differences between measurements performed by each POCT analyzer and the laboratory method (95% confidence interval) were: 0.28% (p<0.005) (0.10-0.44) for DCA and 0.27% (p<0.001) (0.19-0.35) for Afinion. Correlation coefficients were: r=0.973 for DCA, and r=0.991 for Afinion. The mean bias observed by using samples from chronic renal failure patients were 0.2 (range -0.4, 0.4) for DCA and 0.2 (-0.2, 0.5) for Afinion. Imprecision results were: CV=3.1% (high HbA1c) and 2.97% (low HbA1c) for DCA, CV=1.95% (high HbA1c) and 2.66% (low HbA1c) for Afinion. CONCLUSIONS Both POCT analyzers for HbA1c show good correlation with the laboratory method and acceptable precision.

Hemes férricos pentacoordenados e hexacoordenados dos monômeros d nativo e reconstituído da hemoglobina extracelular de Glossoscolex paulistus: estudos espectroscópicos em meio ácido

UV-Vis and fluorescence spectroscopic studies of the native and reconstituted d monomers of Glossoscolex paulistus were performed in acid medium. The coexistence of distinct species shows the complexity of the equilibria. Besides the hexacoordinate low spin hemichrome, with bands at 535 and 565 nm, a pentacoordinate high spin hemichrome is identified by the blue-shifted low intensity Soret band (371 nm) and the LMCT band (643 nm). The pentacoordinate hemichrome must be related to the partial unfolding of the polypeptide.

Estudo espectroscópico do equilíbrio entre espécies hexacoordenada e pentacoordenada dos monômero d nativo e reconstituído da hemoglobina extracelular gigante de Glossoscolex paulistus em meio alcalino

UV-Vis and fluorescence spectroscopic studies of native and reconstituted d monomers from Glossoscolex paulistus were performed in alkaline pH. In addition to the presence of aquomet and hemichrome hexacoordinated species, a pentacoordinated species occurs. This latter species, which presents an imidazolate group of axial histidine as fifth ligand, is identified by the blue-shifted low-intensity Soret band and LMCT band. The pentacoordinated species must occur due to partial polypeptide unfolding.

Hemoglobina extracelular gigante de Glossoscolex paulistus: um extraordinário sistema supramolecular hemoproteico

Giant extracellular hemoglobins are considered the summit of complexity in systems that carry oxygen, constituting an extraordinary model system to the study of hemoproteins. This class includes the hemoglobin of the annelid Glossoscolex paulistus that presents high cooperativity, great oligomeric and redox stabilities and ability of oligomeric reassociation. These properties have motivated evaluations about its utilization as prototype of artificial blood and biosensor. Kinetic studies involving autoxidation and detailed spectroscopic characterizations of its ferrous and ferric species have propitiated information about the structure-activity relationship of this macromolecule. The present review analyzes several biochemical issues, evaluating the state-of-art of this subject.

Síntese e caracterização de copolímeros do tipo ABA para encapsulação de hemoglobina bovina

The use of biopolymers for the development of oxygen carriers has been extensively investigated. In this work, three different ABA triblock copolymers were synthesized and used to encapsulate purified bovine hemoglobin, using a double emulsion technique. The effect of polymer composition, homogenization velocity, and addition of a surfactant, on the protein entrapment was evaluated. These copolymers, which have a hydrophilic block, achieved higher values of encapsulation efficiency than the corresponding homopolymers. The increase in homogenization strength also promoted an increase in encapsulation efficiency. Capsules formation occurred even in the absence of PVA.

Hemocidinas derivadas da hemoglobina: estruturas, propriedades e perspectivas

The increasing incidence of microbial infections, high toxicity, and high level of resistance associated with conventional antibiotics has created a need for new drugs. Antimicrobial peptides (AMPs) constitute a promising alternative and/or an important source of knowledge given their ability to inhibit the growth and/or to kill bacteria, fungi, parasites and/or viruses through mechanisms of action different from those of non-peptide drugs. This review focused on this important class of organic compounds that includes hemocidins resulting from hemoglobin proteolysis in vivo and in vitro or from chemical synthesis, subject of research in foreign and Brazilian laboratories.

Níveis de hemoglobina glicosilada e anomalias cardíacas em fetos de mães com diabetes mellitus

Avaliou-se, prospectivamente, a existência de relação entre o controle glicêmico materno, na primeira metade da gestação, com a ocorrência de anomalias cardíacas fetais, em gestantes com diabetes mellitus. O nível da hemoglobina glicosilada (HbA1c) foi determinado em 127 grávidas, por ocasião da primeira visita pré-natal. Nove eram portadoras de diabetes do tipo I, 77 do tipo II e 41 de Diabetes mellitus gestacional (DMG). Todas foram submetidas a ecocardiografia fetal detalhada na 28ª (±4,1) semana da gestação. Em 31 dos 127 fetos (24,4%) foram detectadas anomalias cardíacas. Em 10 (7,87%), foram diagnosticadas anomalias cardíacas estruturais e, em 21 (16,53%), miocardiopatia hipertrófica como anomalia cardíaca isolada. A média das dosagens de HbA1c no grupo de gestantes sem anomalias cardíacas (5,6%) foi estatisticamente diferente da média das HbA1c do grupo com anomalias (10,1%) (p<0,0001). A elaboração da curva ROC, representando o balanço entre a sensibilidade (92,83%) e a especificidade (98,92%) no diagnóstico das anomalias cardíacas estruturais, com base nos níveis da HbA1c e resultados da ecocardiografia fetal, revelou ponto de corte discriminatório em 7,5%. Em nove dos dez fetos portadores de anomalias cardíacas estruturais, o nível materno da HbA1c era maior do que 7,5%. Não foi observada diferença estatisticamente significante quando se compararam as médias das dosagens de HbA1c dos casos com e sem miocardiopatia hipertrófica, diagnosticada como anomalia isolada (MCPH), nos subgrupos de diabetes tipo II e DMG. Concluem os autores que o nível de HbA1c acima de 7,5% correlaciona-se com o diagnóstico ecocardiográfico de anomalias cardíacas estruturais. Por outro lado, este teste não se mostrou útil para discriminar conceptos portadores de MCHP.

Correlação entre os índices dopplervelocimétricos da veia cava inferior e ducto venoso e a concentração de hemoglobina do cordão em fetos de gestantes isoimunizadas

OBJETIVO: o objetivo principal desse estudo foi verificar se existe correlação entre os índices dopplervelocimétricos da veia cava inferior e do ducto venoso e a concentração sérica da hemoglobina fetal. MÉTODOS: estudo transversal e prospectivo, realizado entre janeiro de 1998 e junho de 2001. Foram acompanhadas 31 gestantes isoimunizadas com resultado do teste de Coombs indireto maior que 1:8, que foram submetidas à pesquisa de hemólise fetal. Quando foram indicadas as transfusões intra-uterinas intravasculares, a hemoglobina do cordão foi dosada no início do procedimento. Nos demais casos, a hemoglobina do cordão foi mensurada no momento do parto, sempre por cesariana eletiva. Obteve-se um total de 74 procedimentos estudados, definidos como sendo cada transfusão intra-uterina precedida pela dopplervelocimetria venosa. A mensuração da concentração da hemoglobina fetal foi realizada no Hemocue® (B-Hemoglobin Photometer Hemocue AB; Angelholm, Sweden), dispositivo usado para a determinação quantitativa de hemoglobina no sangue. A dopplervelocimetria da cava inferior e do ducto venoso foi realizada antecedendo a coleta do sangue fetal, sempre em intervalo de tempo inferior a 24 horas. Os índices dopplervelocimétricos estudados foram o índice de pulsatilidade para veias (IPV), o índice do pico de velocidade para veias (IPVV) e a relação entre o pico de velocidade durante a fase de contração atrial e o pico de velocidade na sístole ventricular (relação CA/SV ou índice de pré-carga), na veia cava inferior, e o IPV, IPVV e a relação entre os picos de velocidade da sístole ventricular e da contração atrial (relação SV/CA), no ducto venoso. Foi realizado estudo de correlação entre a dopplervelocimetria da veia cava inferior e do ducto venoso e a hemoglobina do cordão, pela técnica de regressão linear simples. Realizou-se também estudo de associação entre os índices dopplervelocimétricos do compartimento venoso e a hemoglobina fetal inferior a 10 g/dL, por meio do teste do c². RESULTADOS: foi efetuado um total de 74 procedimentos. Em 23 desses a hemoglobina fetal era inferior a 7,0 g/dL. Houve correlação negativa significativa entre todos os índices dopplerfluxométricos estudados e a concentração de hemoglobina fetal (p<0,05), ou seja, os valores mais elevados dos índices dopplervelocimétricos foram observados nos fetos com anemia mais acentuada. Os fetos com nível de hemoglobina inferior ou igual a 10,0 g/dL apresentaram valores dos índices dopplervelocimétricos da cava e do ducto venoso acima do percentil 95 para a idade gestacional. CONCLUSÕES: a dopplervelocimetria da cava inferior e do ducto venoso pode ser utilizada como marcador não invasivo da anemia fetal grave.