Anna L. Soares

High-sensitivity C-reactive protein in subjects with type 2 diabetes mellitus and/or high blood pressure

Type 2 diabetes mellitus (DM2) and high blood pressure (HBP) may contribute to the development of cardiovascular disease, and inflammation may be an important factor in these diseases. In the present study, plasma levels of high-sensitivity C-reactive protein (hs-CRP) were measured in subjects with DM2 and/or HBP and compared to those of normal subjects. Eighty-nine subjects were analyzed for hs-CRP, including 13 normotensive patients with DM2, 17 patients with HBP, 34 hypertensive patients with DM2 (DM2+HBP) and 25 normal subjects. The plasma hs-CRP levels were significantly lower in the controls than in the HBP+DM2 group (p < 0.05). DM2 associated with HBP was also correlated with increased plasma hs-CRP levels (n = 89, r = 0.25, p = 0.0162). Only hypertensive patients with DM2 had higher levels of hs-CRP, a circulating inflammatory marker, than normal subjects. This finding suggests that patients with two associated diseases have a more active inflammatory state.

Elevated plasma factor VIII and von Willebrand factor in women with type 2 diabetes: inflammatory reaction, endothelial perturbation or else?

The association between type 2 diabetes and cardiovascular disease is long recognized. Although perturbations of haemostatic markers have been shown to be associated with macrovascular disease in patients with type 2 diabetes, it is unclear whether these are primarily due to endothelial dysfunction or a result of inflammation. The present study was undertaken to elucidate whether elevated levels of factor VIII (FVIII) and von Willebrand factor (vWF) in women with type 2 diabetes represent endothelial dysfunction, inflammation or an alternate mechanism. Sixty-four women with type 2 diabetes were evaluated using ultrasonography Doppler for carotid intima–media thickness (IMT) and were classified as group A – having no (<1 mm), group B – mild (≥1 mm and no plaque) and group C – moderate (≥1 mm and presence of plaque and stenosis) macrovascular disease. Several haemostatic markers including, FVIII, vWF and fibrinogen were assessed. In addition, thrombomodulin, a marker for endothelial damage, and high-sensitivity C-reactive protein (hsCRP), an inflammatory marker, were also measured. A significant association of elevated FVIII was found in group B and C patients (i.e. patients with IMT ≥1 mm and with plaque). Elevated fibrinogen and vWF levels were also found but confined to group C patients. No significant difference among subgroups was found for any other variable evaluated (hsCRP, thrombomodulin and FVII). In conclusion, plasma FVIII levels are elevated in women with type 2 diabetes and macrovascular disease. It also appears that this is not mediated by inflammation or endothelial injury and is likely to be due to an alternate mechanism.

PAI-1 and D-dimer in type 2 diabetic women with asymptomatic macrovascular disease assessed by carotid Doppler.

Asymptomatic diabetic patients with different degrees of macrovascular complications can present different hemostatic changes. At this study, plasminogen activator inhibitor-1 (PAI-1) and D-dimer were evaluated in 12 women without diabetes and 64 type 2 diabetic women. All patients were classified into 3 different categories according to the carotid intima-media thickness (IMT) assessed by Doppler: 25 with <1 mm (normal), 15 with >1 mm and without plaque (intermediate), and 24 with stenosis lower than 50% of the vessel lumen (plaque). The results showed increased plasma D-dimer in type 2 diabetic women with carotid plaque when compared to the other groups. High levels of PAI-1 were observed in all the 3 groups of diabetic women when compared to women without diabetes. Our results suggest that high levels of PAI-1 in type 2 diabetic women are only associated with diabetes and are not associated with macrovascular progression; however, it seems that D-dimer plasma levels are associated with carotid plaque.

Plasma Total Homocysteine Levels and Methylenetetrahydrofolate Reductase Gene Polymorphism in Patients with Type 2 Diabetes Mellitus

Background: Thrombotic episodes account for approximately 80% of deaths in type 2 diabetic patients. Hyperhomocysteinaemia is a well recognized independent risk factor for atherosclerosis and thromboembolism. Increased homocysteine levels may occur due to a number of factors including inherited gene polymorphism of methylenetetrahydrofolate reductase (MTHFR) C677T. Here, we evaluate plas- ma total homocysteine (tHcy) levels and frequency of the MTHFR C677T gene polymorphism in asymptomatic healthy volunteers and type 2 diabetic patients with hypertension but without nephropathy. We have also investigated the relationship between tHcy levels and the presence of MTHFR C677T gene polymorphism. Methods: Plasma tHcy levels and MTHFR C677T genotype were investigated in a total of 53 subjects. These included asymptomatic healthy volunteers (n = 16), patients with type 2 diabetes (n = 7), subjects with hypertension (n = 12) and patients with both type 2 diabetes and hypertension (n = 18). Renal function, serum lipids and other metabolites were also assessed. Results: There was no significant difference in tHcy levels between the groups studied. The frequency of MTHFR C677T gene polymorphism observed was similar to that obtained for the general Brazilian population. In patients with type 2 diabetes and hypertension but without impaired renal function, we observed no meaningful correlation between increased tHcy levels and the presence of MTHFR C677T gene polymorphism. Conclusions: Type 2 diabetics who are homozygous or heterozygous for the MTHFR C677T gene polymorphism showed normal tHcy levels. Our results further suggest that diabetes without an associated adverse risk profile is not an independent correlate of increased tHcy levels.

Correlação entre os níveis plasmáticos de apolipoproteínas A-I e B e o perfil lipídico em indivíduos com e sem diabetes mellitus tipo 2 e hipertensão arterial

Background: Dyslipidemias, diabetes mellitus (DM), high blood pressure are important factors for development of the coronary artery disease (CAD), principal cause of death in the world. Several studies have demonstrated positive correlation between both LDL-C high plasma levels and HDL-C low concentrations and increased risk for cardiovas- cular diseases. Objectives: To establish the possible correlation between lipids, lipoproteins, apolipoproteins A-I and B, Lipoprotein(a) and microalbuminuria in subjects with and without diabetes mellitus type 2 and high blood pressure. Material and Method: The subjects, with age range from 40 to 65 years, were divided into five groups: 1. control (normal subjects, n = 16); 2. HAS (subjects with high blood pressure, n = 12); 3. DM (normotensive and normoalbuminuric patients with diabetes mellitus type 2, n = 7); 4. DM + HASnAlb (hypertensive and normoalbuminuric patients with diabetes mellitus type 2, n = 18) and 5. DM + HASmAlb (hypertensive and microalbuminuric patients with diabetes mellitus type 2, n = 9). Results: Concerning to the lipid profile statistical differences were observed for LDL-C between the average of the group 5 compared to the averages of the other groups; for triglycerides among the groups 2, 4 and 5 compared to the group 1. For the total cholesterol, HDL-C, Lp(a) and Apo A-I no significant difference was observed among the groups. Average values for Apo B for groups 4 and 5 presented significant difference compared to group 1. Positive correlation was observed between LDL-C and Apo B (r = 0,684); p < 0,0001) and HDL-C and Apo A-I (r = 0,374; p = 0,003), according to literature. Conclusion: Among all parameters assessment, LDL-C was the unique one that showed significant differences between the group 5, whose participants present both alterations (DM and HAS) besides the microalbuminuria, related to other groups studied. This comes to suggest that the association between the two diseases allied to microalbuminuria may contribute to the dyslipidemia aggravation.

Alterações do sistema hemostático nos pacientes com diabetes melito tipo 2

Diabetes has acquired an epidemic character due to the large increase in the number of individuals affected over recent decades. Diabetes-related mortality is associated with thrombotic events, especially cardiovascular. In general, patients with diabetes present symptoms of hypercoagulability and hypofibrinolysis. However, the mechanisms that trigger hemostatic abnormalities in diabetic patients are not clear. The aim of this paper was to address the most frequent changes of the hemostatic system in diabetic patients described in the literature. Diabetics have abnormalities of the endothelium, platelets, clotting factors, natural anticoagulants and the fibrinolytic system; all these changes are directly and/or indirectly caused by hyperglycemia. Thus, analytes such as von Willebrand factor, factor VIII, fibrinogen and D-dimer are markers that should be interpreted differently in diabetic patients. Laboratory evidence of hemostatic abnormalities in diabetic patients supports clinical observations that diabetes is a state of hypercoagulability and hypofibrinolysis. Strategies for clinical intervention and medications are not well established considering the results of the hemostatic markers.

Velocidade de hemossedimentação: comparação entre o método Microtest X (microssedimentação) e o método de referência Westergren

The erythrocyte sedimentation rate (ESR) is an easy and common routine laboratory technique. Over the last few decades, new methods have been proposed to reduce biological sample manipulation, to measure rates using an automated system, to optimize the workflow and to reduce blood volume from venipuncture. The aim of this study was to compare the efficacy of the Microtest X technique with the original Westergren method. EDTA blood samples of 25 subjects were submitted to both methodologies. A positive correlation (p<0.0001; R2=0.809) was found as was agreement with Bland-Altman analysis. In conclusion, the Microtest X technique allows precise and accurate measurements of ESR and can be used to replace the Westergren method. The benefits of Microtest X are the use of very small volumes of EDTA samples, it reduces sample manipulation avoiding occupational hazards and markedly reduces the analytical time.

Clinical and Molecular Features of Patients with Congenital Disorders of Glycosylation in Brazil

Introduction: Congenital Disorders of Glycosylation are a group of genetic disorders due to abnormal glycosylation of glycoproteins and glycolipids. Based on isoelectric focusing of plasma transferrin results, CDG are classified in two groups: CDG-I and CDG-II. While the diagnosis of PMM2-CDG (formerly CDG-Ia) and PMI-CDG (formerly CDG-Ib) is made by demonstration of the enzyme deficiency or by gene sequencing, the diagnosis of the other CDG is not easily performed. Psychomotor delay/mental retardation, hypotonia, seizures, ataxia, cerebellar atrophy, strabismus, inverted nipples, lipodystrophy, and stroke-like episodes characterize PMM2-CDG, by far the most common CDG. There is almost no information available in the literature on the frequency of CDG in patients with psychomotor delay/ mental retardation. Patients and methods: We performed transferrin isoelectric focusing in 2619 patients who had psychomotor delay/mental retardation associated with other symptoms suggestive of CDG. Determination of leukocyte phosphomannomutase and phosphomannoseisomerase activities and PMM2 gene sequencing was performed in selected patients. Results: We found 32 affected patients (26 CDG-I and 6-CDG-II). CDG-I group: The most prevalent PMM2- CDG clinical symptoms were those expected. We identified two novel mutations: p.G79V and p.R21W. Non-PMM2, non-PMI-CDG showed more frequently coagulopathy, hypotonia, cerebellar atrophy, and cryptorchidism/micropenis. Early deaths were found exclusively in this group. Ataxia, strabismus, elevated blood FSH and LH levels were more frequent in PMM2-CDG patients. CDG-II group: four out of six patients presented cutis laxa, seizures, large fontanel, facial dysmorphism, and non-lissencephalic cortical dysplasia. Hip luxation was present in three patients, and hydronephrosis in one. The other two patients had heterogeneous features. Conclusions: We determined the frequency of CDG in a selected Brazilian cohort with symptoms suggestive of CDG as 1.2%(CDG-I ~ 1.0% and CDG-II ~ 0.2%), and identified two novel mutations in the PMM2 gene.

Avaliação da incidência das mutações G1691A no gene do fator V (fator V Leiden) e G20210A no gene da protrombina em pacientes com diabetes mellitus tipo 2

Em virtude da alta prevalencia de Diabetes mellitus tipo 2 (DM2) na populacao mundial e da alta taxa de mortalidade decorrente de eventos tromboticos, e de extrema importância o conhecimento das alteracoes no sistema hemostatico em pacientes portadores deste disturbio. A mutacao no gene do fator V (G1691A - fator V Leiden) em heterozigose ou homozigose confere aos portadores o fenotipo de resistencia a proteina C ativada, situacao que aumenta em sete vezes o risco de desenvolver uma trombose. A mutacao G20210A no gene da protrombina resulta no quadro de hiperprotrombinemia, aumentando o risco de trombose em tres vezes. A pesquisa dessas mutacoes de interesse em trombofilia e de grande relevância considerando que a presenca das mesmas pode exacerbar o estado de hipercoagulabilidade acelerando as complicacoes no diabetes. O presente estudo teve como objetivo avaliar a incidencia dessas mutacoes em individuos higidos (Controle, n=16), pacientes com DM2 (n=7), com hipertensao (HAS, n=12) e com DM2+HAS (n=18), atraves da tecnica de PCR-RFLP. As frequencias encontradas nos grupos estudados foram baixas e similares aquelas observadas na populacao brasileira em geral. Nao foi possivel estabelecer correlacao entre a presenca da mutacao e caracteristicas especificas de cada grupo. Dessa forma, ainda nao esta claro se ha ou nao uma maior prevalencia dessas mutacoes em individuos diabeticos e se a presenca das mesmas contribui para o aumento do risco de desenvolver trombose nesses individuos, sendo necessario estudos mais amplos para a elucidacao da questao.

Murine schistosomiasis mansoni: process of blood coagulation at pre-patent, acute and chronic phases, and consequence of chemotherapeutic cure on the reversion of changes.

The present study aims to elucidate in a sequential manner the changes of the blood coagulation process at different phases of experimental schistosomiasis, comprising the pre-patent, acute, intermediate and chronic phases, and the effect of chemotherapeutic cure, at the acute and chronic phases, on reversion of changes related to the coagulation factors. Mice were infected with Schistosoma mansoni cercariae, and were divided into four groups. Blood samples from these groups were collected 32, 70, 100, and 140 days after infection, corresponding to the pre-patent, acute, intermediate and chronic phases, respectively. Simultaneously, other infected groups were given oxamniquine, 70 and 140 days after infection. At the same time as blood collection from infected and/or treated animal groups, other uninfected control animal groups were punctured and maintained under the same conditions as the infected animals. The vitamin-K-dependent clotting factors were found to be more sensitive to infection at different phases, while factors VIII and XI presented hyperactivity. Results obtained 90 days after chemotherapeutic treatment with oxamniquine, administered at the acute and chronic phases, presented noticeable reversion of the main alterations in the coagulation mechanism. The present study provides unquestionable data on the development of hemostatic changes throughout the course of S. mansoni infection.