Interaction between oral estrogen plus progestogen therapy and ABO blood groups on coagulation activation in postmenopausal women.

OBJECTIVE: The aim of this study was to investigate the effect of postmenopausal hormone therapy on coagulation and whether this effect differs according to ABO blood groups. METHODS: This was a prospective observational study to evaluate factor VIII (FVIII) activity, factor von Willebrand (vWF), and D-dimer (D-Di) levels and ABO blood groups in 61 postmenopausal women using oral estrogen plus progestogen therapy (EPT; 2 mg estradiol + 1 mg norethisterone acetate) for 3 months and in 101 women not using EPT. After 3 months, all eligible women who had completed the treatment scheme proposed for the EPT group or those who opted to participate but had not undergone EPT had a blood sample collected for analysis. RESULTS: Significant differences were observed in FVIII activity and vWF levels in the control group between those carrying group O and non-group O blood. For EPT users, significant differences were observed for FVIII activity, vWF, and D-Di levels. After a multivariate regression analysis, FVIII activity and ABO blood groups were independently associated with vWF levels, whereas interaction between ABO blood groups and EPT were independently associated with FVIII activity. Besides diabetes, the ABO × EPT interaction was also noted to be independently associated with D-Di levels. CONCLUSIONS: These findings suggest an interactive effect between oral EPT and non-O blood groups, contributing to the mechanism by which estrogen triggers the hypercoagulability state and increased risk for venous thrombosis in women undergoing oral EPT.

Epidermal growth factor receptors, testosterone levels and parotid gland changes in rats infected with Trypanosoma cruzi.

It has been demonstrated that parotid glands of rats infected with Trypanosoma cruzi present severe histological alterations; changes include reduction in density and volume of the acini and duct systems and an increase in connective tissue. We evaluated the association between morphological changes in parotid glands, circulating testosterone levels and epidermal growth factor receptor (EGF-R) expression in experimental Chagas disease in rats. Animals at 18 days of infection (acute phase) showed a significant decrease in body weight, serum testosterone levels and EGF-R expression in the parotid gland compared with a control group. Since decreases in body weight could lead to a reduction in circulating testosterone concentration, we believe that the reduction in EGF-R expression in parotid glands of infected rats is due to alterations in testosterone levels and atrophy of parotid glands is caused by changes in EGF-R expression. Additionally, at 50 days (chronic phase) of infection parotid glands showed a normal histological aspect likely due to the normalization of the body weight. These findings suggest that the testosterone-EGF-R axis is involved in the histological changes.

Epidermal growth factor receptors, testosterone levels and parotid gland changes in rats infected with Trypanosoma cruzi

It has been demonstrated that parotid glands of rats infected with Trypanosoma cruzi present severe histological alterations; changes include reduction in density and volume of the acini and duct systems and an increase in connective tissue. We evaluated the association between morphological changes in parotid glands, circulating testosterone levels and epidermal growth factor receptor (EGF-R) expression in experimental Chagas disease in rats. Animals at 18 days of infection (acute phase) showed a significant decrease in body weight, serum testosterone levels and EGF-R expression in the parotid gland compared with a control group. Since decreases in body weight could lead to a reduction in circulating testosterone concentration, we believe that the reduction in EGF-R expression in parotid glands of infected rats is due to alterations in testosterone levels and atrophy of parotid glands is caused by changes in EGF-R expression. Additionally, at 50 days (chronic phase) of infection parotid glands showed a normal histological aspect likely due to the normalization of the body weight. These findings suggest that the testosterone-EGF-R axis is involved in the histological changes.

Frequência do polimorfismo da glicoproteína IIIa de plaquetas (PlA2) em mulheres com diabetes mellitus tipo 2 / Platelet glycoprotein IIIa polymorphism (PlA2) frequency in Type 2 diabetic women

O polimorfismo da glicoproteína IIIa de plaquetas está associado a um aumento no risco de doenças arteriais coronarianas. Mulheres com diabetes mellitus tipo 2 apresentam um aumento de cinco vezes no risco para doenças arteriais coronarianas quando comparadas com mulheres não-diabéticas. O objetivo do presente estudo foi verificar a frequência do polimorfismo da glicoproteína IIIa (PlA2) em mulheres com diabetes mellitus tipo 2 e comparar com a frequência descrita na literatura. A análise do polimorfismo PlA2 foi realizada para 62 mulheres com diabetes mellitus tipo 2 através da reação em cadeia da polimerase seguida de análise do polimorfismo de tamanho de fragmento de restrição (PCR-RFLP). As frequências observadas foram 81 por cento para PlA1A1, 18 por cento para PlA1A2 e 1 por cento para PlA2A2. Não houve diferença significativa entre as frequências observadas e as frequências descritas na literatura. Nossos resultados sugerem que a frequência do polimorfismo PlA2 em mulheres com diabetes mellitus tipo 2 é a mesma observada na população em geral.

The platelet glycoprotein IIIa polymorphism is associated to an increased risk of coronary heart disease. Type 2 diabetic women present a fivefold higher risk of coronary heart disease compared to non-diabetic women. The aim of this study was to verify the frequency of the glycoprotein IIIa polymorphism (PlA2) in type 2 diabetic women and compare this result with the frequency reported for the general population. The PlA polymorphisms of 62 type 2 diabetic women were determined by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). The resulting frequencies were 81 percent for PlA1A1, 18 percent for PlA1A2 and 1 percent for PlA2A2. There was no significant difference between observed frequencies and the frequencies described in the literature. Our results suggest that the frequency of the glycoprotein IIIa polymorphism, PlA2, in type 2 diabetic women is similar to that observed in general population

Assessment of hypercoagulability markers and lipid levels in postmenopausal women undergoing either oral or transdermal hormone replacement therapy.

BACKGROUND: This study investigated the effect of either oral or transdermal hormone replacement therapy (HRT) on haemostatic, fibrinolytic and lipid profiles in a group of Brazilian women 3 months after beginning treatment by comparing these results with those obtained immediately before HRT. METHODS: Plasma levels of TAT, DDi, F1+2, PC, PS, AT, PAI-1 and serum lipids were determined in blood samples collected from 24 women undergoing oral HRT and from 11 women undergoing transdermal HRT. RESULTS: Significant increases in DDi and F1+2 plasma levels were observed after 3 months of oral HRT, while PS levels decreased. After transdermal HRT, a significant decrease was observed only for AT levels. CONCLUSION: After 3 months of oral HRT and in the absence of major genetic and acquired risk factors, women displayed a predisposition for activation of blood coagulation, and an increased activity of the fibrinolytic system. Oral HRT seemed to be more effective in predisposing haemostatic changes as compared to transdermal.

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.

Effect of oral hormone replacement therapy on plasma homocysteine levels.

BACKGROUND: Various inherited or acquired conditions can lead to mild or severe hyperhomocysteinemia, which has toxic effects on the vascular endothelium. It has been reported that hormone replacement therapy is associated with decreased homocysteine plasma levels, but this is still a controversial issue. PURPOSE: To compare homocysteine plasma levels in women before and after 3 months of oral hormone replacement therapy. METHODS: Twenty-four women were selected to take part in the study. Blood samples were collected immediately before hormone replacement therapy (cyclic association of 2 mg of estradiol valerate and 1 mg of cyproterone acetate) and three months after the beginning of hormone replacement therapy. Samples collected before hormone replacement therapy were used as controls. Plasma homocysteine levels and the presence of C677T mutation in the methylene tetrahydrofolate reductase gene were evaluated in all participants. RESULTS: The methylene tetrahydrofolate reductase gene mutation was detected in 8 women (33.3%) in heterozygosis, in 3 (12.5%) in homozygosis, and 13 women (54.2%) did not present the mutation. No significant differences were observed in homocysteine levels before and after three months of oral hormone replacement therapy, regardless of the C677T genotype. CONCLUSIONS: The results obtained indicate that homocysteine plasma levels are not affected after three months of oral hormone replacement therapy.

Long-term protective immunity induced against Trypanosoma cruzi infection after vaccination with recombinant adenoviruses encoding amastigote surface protein-2 and trans-sialidase.

Protection against protozoan parasite Trypanosoma cruzi has been shown to be dependent on the induction of type 1 immune responses. Replication-deficient human type 5 recombinant adenoviruses have an unsurpassed ability to induce type 1 immune responses. Thus, we constructed two type 5 recombinant adenoviruses encoding parasite antigens trans-sialidase (rAdTS) and amastigote surface protein-2 (rAdASP2). Both antigens were genetically engineered to secrete recombinant products in order to induce both optimal antibody and T cell responses. Immunizations of mice with rAdASP2 and rAdTS induced high levels of serum antibodies specific for their recombinant products. In addition, both recombinant viruses were able to elicit a biased helper T cell type 1 (Th1) cellular immune response and a substantial CD8+ T cell-mediated immune response. Moreover, individual immunization with rAdASP2 or rAdTS induced high levels of protection against a challenge with live parasites. CD8+ T cells mediated, at least in part, such protection. Furthermore, when combined in the same inoculum, rAdTS plus rAdASP2 induced complete protection in all animals tested, even when challenges were performed 14 weeks after the last immunization. Taking together, these results show that recombinant adenoviruses expressing TS and ASP-2 antigens of T. cruzi are interesting candidates for the development of a vaccine against Chagas' disease.

Factor V Leiden and increased risk for arterial thrombotic disease in young Brazilian patients.

The contribution of mutations in the prothrombin (FII G20210A), methylenetetrahydrofolate reductase (C677T) genes and factor V Leiden (FVL) to the pathogenesis of arterial thrombosis remains controversial. In this study, these polymorphisms were investigated by polymerase chain reaction-restriction fragment length polymorphism in a group of 53 patients that presented arterial thrombosis other than myocardial infarction as a first thrombotic event and 275 control subjects living in the state of Minas Gerais, Brazil. Odds ratio (OR) and chi tests were applied for statistical comparisons. Similar frequencies were detected among patients and control subjects for the C677T mutation. The 20210A mutation was present in 3.6% of the control subjects but was not detected among ischemic stroke patients. Significant differences were detected only for factor V Leiden (odds ratio 7.11; 95% confidence interval 1.55-32.73). Our data indicate that, among these genetic factors, factor V Leiden was identified as an important risk factor for arterial thrombosis in this group of patients. In addition, our results indicate regional differences in the incidence of these genetic factors in Brazil, as compared to the incidences reported in other studies.

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 / Incidence of G1691A factor V gene (factor V Leiden) and G20210A prothrombin gene mutations in type 2 diabetes patients

Em virtude da alta prevalência de Diabetes mellitus tipo 2 (DM2) na população mundial e da alta taxa de mortalidade decorrente de eventos trombóticos, é de extrema importância o conhecimento das alterações no sistema hemostático em pacientes portadores deste distúrbio. A mutação no gene do fator V (G1691A - fator V Leiden) em heterozigose ou homozigose confere aos portadores o fenótipo de resistência à proteína C ativada, situação que aumenta em sete vezes o risco de desenvolver uma trombose. A mutação G20210A no gene da protrombina resulta no quadro de hiperprotrombinemia, aumentando o risco de trombose em três vezes. A pesquisa dessas mutações de interesse em trombofilia é de grande relevância considerando que a presença das mesmas pode exacerbar o estado de hipercoagulabilidade acelerando as complicações no diabetes. O presente estudo teve como objetivo avaliar a incidência dessas mutações em indivíduos hígidos (Controle, n=16), pacientes com DM2 (n=7), com hipertensão (HAS, n=12) e com DM2+HAS (n=18), através da técnica de PCR-RFLP. As freqüências encontradas nos grupos estudados foram baixas e similares àquelas observadas na população brasileira em geral. Não foi possível estabelecer correlação entre a presença da mutação e características específicas de cada grupo. Dessa forma, ainda não está claro se há ou não uma maior prevalência dessas mutações em indivíduos diabéticos e se a presença das mesmas contribui para o aumento do risco de desenvolver trombose nesses indivíduos, sendo necessário estudos mais amplos para a elucidação da questão.

Because of the high prevalence of type 2 diabetes mellitus (DM2) worldwide and the high mortality rate due to thrombotic events, it is extremely important to know about changes in the hemostatic system of such patients. Factor V mutation (G1691A - factor V Leiden) in either heterozygosis or homozygosis confers the activated protein C resistant phenotype, which increases the risk of thrombotic events by a factor of seven. The G20210A mutation of the prothrombin gene results in hyperprothrombinemia, increasing the risk of thrombotic events by a multiple of three. These mutations are of great relevance considering that the presence of one or both can contribute to a hypercoagulability state accelerating complications in diabetes. The aim of this study was to evaluate the incidence of these mutations in controls (n=16), DM2 subjects (n=7), hypertensive subjects (HAS) (n=12) and DM2+HAS subjects (n=18). The frequencies found were low and similar to those observed in the Brazilian population in general. It was not possible to establish any correlation among mutations and specific features of each group. It is still not clear if there is or not a higher prevalence of these mutations in diabetic individuals or if the mutation contributes to an increase in the risk of thrombotic events in these individuals. Further studies involving a larger number of patients are necessary in order to answer these questions.

Activity of a paromomycin hydrophilic formulation for topical treatment of infections by Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis.

Studies on in vitro skin permeation and in vivo anti-leishmanial activity in mice experimentally infected with Leishmania (Leishmania) major pointed out to the potential of a new paromomycin (PA) formulation (hydrophilic gel) for treatment of cutaneous leishmaniasis (CL). In this study, the activity of this formulation was evaluated in animals experimentally infected by Leishmania species that prevail in the New World. PA gel activity was compared to antimony treatment, since it is still the first choice treatment to the different clinical forms of leishmaniasis. The topical treatment activity with 10% PA gel in BALB/c mice infected by Leishmania (Leishmania) amazonensis was higher than that observed for parenteral antimony treatment, while the efficacy of these two regimes in hamsters infected by Leishmania (Viannia) braziliensis was similar. These results suggest that this formulation could be suitable for clinical studies and may represent an alternative novel formulation for topical treatment of CL.

Impaired production of proinflammatory cytokines and host resistance to acute infection with Trypanosoma cruzi in mice lacking functional myeloid differentiation factor 88.

Studies performed in vitro suggest that activation of Toll-like receptors (TLRs) by parasite-derived molecules may initiate inflammatory responses and host innate defense mechanisms against Trypanosoma cruzi. Here, we evaluated the impact of TLR2 and myeloid differentiation factor 88 (MyD88) deficiencies in host resistance to infection with T. cruzi. Our results show that macrophages derived from TLR2 (-/-) and MyD88(-/-) mice are less responsive to GPI-mucin derived from T. cruzi trypomastigotes and parasites. In contrast, the same cells from TLR2(-/-) still produce TNF-alpha, IL-12, and reactive nitrogen intermediates (RNI) upon exposure to live T. cruzi trypomastigotes. Consistently, we show that TLR2(-/-) mice mount a robust proinflammatory cytokine response as well as RNI production during the acute phase of infection with T. cruzi parasites. Further, deletion of the functional TLR2 gene had no major impact on parasitemia nor on mortality. In contrast, the MyD88(-/-) mice had a diminished cytokine response and RNI production upon acute infection with T. cruzi. More importantly, we show that MyD88(-/-) mice are more susceptible to infection with T. cruzi as indicated by the higher parasitemia and accelerated mortality, as compared with the wild-type mice. Together, our results indicate that T. cruzi parasites elicit an alternative inflammatory pathway independent of TLR2. This pathway is partially dependent on MyD88 and necessary for mounting optimal inflammatory and RNI responses that control T. cruzi replication during the early stages of infection.

Glycosylphosphatidylinositol-anchored mucin-like glycoproteins from Trypanosoma cruzi bind to CD1d but do not elicit dominant innate or adaptive immune responses via the CD1d/NKT cell pathway.

It has been proposed that self and protozoan-derived GPI anchors are natural ligands of CD1d. In this study, we investigated the ability of GPI anchors from Trypanosoma cruzi to bind to CD1d and mediate activation of NKT cells. We observed that GPI-anchored mucin-like glycoproteins (GPI mucins), glycoinositolphospholipids (GIPLs), and their phosphatidylinositol moieties bind to rCD1d and inhibit the stimulation of a NKT hybridoma by the alpha-galactosylceramide-CD1 complex. However, these GPI anchors and related structures were unable to activate NKT cells in vitro or in vivo. We found that high titers of Ab anti-GPI mucins, but not anti-GIPLs, were detected in sera from wild-type as well as in TAP1(-/-), CD1d(-/-), and MHC class II(-/-) mice after immunization. However, T-dependent anti-GPI mucin Ab isotypes, such as IgG1, IgG2a, IgG2b, and IgG3, were absent on MHC class II(-/-), but were conserved in CD1d(-/-) and TAP1(-/-) mice. Furthermore, we found that CD1d(-/-) mice presented a robust cytokine as well as anti-GPI mucins and anti-GIPL Ab responses, upon infection with T. cruzi parasites. These results indicate that, despite binding to CD1d, GPI mucins and related structures expressed by T. cruzi appear not to evoke dominant CD1d-restricted immune responses in vivo. In contrast, MHC class II is critical for the production of the major Ig G isotypes against GPI mucins from T. cruzi parasites.

Gestao compartilhada da educacao: a experiencia Catarinense

Objetiva socializar algumas experiencias de gestao compartilhada que a Associacao dos Administradores Escolares de Santa Catarina (AAESC) vem desenvolvendo em diversas regioes, em parceria com a Secretaria de Estado da Educacao, secretarias municipais da educacao, universidades e outros segmentos representativas da sociedade civil, bem como pretende expressar, com clareza, o envolvimento da AAESC com a qualidade da escola publica, tendo como mediacao a gestao compartilhadaf, permitindo que tanto a associacao como os educadores catarinenses possam propor alternativas praticas de administracao do projeto pedagogico. Inicia com um breve historico da AAESC, seguido da discussao dos pressupostos teoricos preliminares e da apresentacao das experiencias em andamento; ao final, sao feitas algumas consideracoes gerais.

Xenodiagnóstico artificial "post-mortem" em chagásicos crônicos / Post-mortem artificial xenodiagnosis in chronic chagasic patients

A fim de obter metodologias que permitam estabelecer, com segurança, o diagnóstico "post-mortem" da infecçäo chagásica, adaptou-se o xenodiagnóstico artifical a necropsiados com diferentes tempos de óbito. O teste foi positivo em três (30%) de dez chagásicos autopsiados. O tempo decorrido entre o êxito letal e o início do repasto pelos triatomíneos destes chagásicos foi de duas horas, duas horas e quinze minutos e sete horas, respectivamente. Discutem-se os fatores que podem explicar a sobrevivência do Trypanosoma cruzi no hospedeiro morto bem como as aplicaçöes práticas do achado