Apresentação e desfecho da febre reumática em uma série de casos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Evolution of subpatent parasitaemia in Trypanosoma cruzi chronically infected mice with the help of a cyclophosphamide amplification transfer assay

Avaliamos o potencial do ensaio clássico de subinoculação, modificado pelo tratamento com ciclofosfamida dos animais receptores, na detecção de parasitemias ocultas em camundongos com in-fecção crônica pelo Trypanosoma cruzi. O ensaio, além de simples, mostrou ter uma alta sensibilidade; assim, utilizando-se parasitas da fase aguda, o tratamento com ciclofosfamida revelou parasitemias em 53,8% dos animais infectados com um tripanosoma da cepa y, e em 20% dos animais infectados com um tripanosoma da cepa CL. O tratamento com ciclofosfamida aumentou a sensibilidade do ensaio de subinoculação nas infecções pela cepa CL, e resultou em igual sensibilidade quando utilizada a cepa Y. Nos camundongos de fase crônica, obtidos a partir de diversos esquemas de imunoprofilaxia (BCG, soro de camundongo imune) ou quimioterapia, o ensaio revelou parasitemias ocultas em 99% dos animais. Auxiliados pelo método da subinoculação-ciclofosfamida estudamos no espaço de um ano a evolução das parasitemias ocultas em um grupo de camundongos infectados que sobreviveram à fase aguda pelo tratamento com Benzonidazol. O ensaio revelou parasitemias ocultas em 100% dos animais. Entretanto, padrões contínuos e discontinuos de positividade puderam ser detectados.

Serum iron and plasma fibrinogen concentrations as indicators of systemic inflammatory diseases in horses

Background: Detection of systemic inflammation, which is important for proper diagnosis and prompt treatment, can be challenging.Hypothesis: Measurement of plasma iron concentration is a sensitive method for detecting systemic inflammation in horses compared with measurements of plasma Fibrinogen concentration, a traditional marker for inflammation in the horse.Animals: Ninety-seven horses hospitalized with diseases causing systemic inflammation, 22 horses with localized inflammation, and 12 clinically normal horses were included in this study.Methods: A retrospective study was made on hospitalized horses that had both plasma iron and fibrinogen concentrations measured on hospital admission.Results: Plasma iron concentration was lower in horses with systemic inflammation (64 +/- 45 mu g/dL) than the reference interval minimum (105 mu g/dL) and were significantly lower (P = .001) than the value in a group of horses with local inflammation (123 +/- 45 mu g/dL) and in healthy transported horses (143 +/- 29 mu g/dL). Low plasma iron and high fibrinogen concentrations were both sensitive indicators of systemic inflammation in horses with sensitivity of 90 and 82%, respectively. There was a similar correlation between either continued decreases in iron concentration (R-sp of 0.239) or increases in fibrinogen concentration (R-sp of 0.280) during hospitalization and a worse prognosis.Conclusions and Clinical Importance: Measurement of plasma iron concentration better reflected acute inflammation than did fibrinogen concentration.

Experimental paracoccidioidomycosis in hamsters (Mesocricetus auratus): Gestational interactions

Paracoccidioides brasiliensis is a dimorphic fungus presenting specific steroid hormone receptors, both in the yeast and mycelial forms and estrogen inhibits the transition from mycelium to yeast. In the acute phase, the disease occurs with equal frequency in both sexes but in adults, females are spared. Placental fungal infection has been reported, but references to fetal infection have not been confirmed. We used 78 Syrian female hamsters divided into 3 groups: GI consisted of 30 infected mated females, GII of 20 infected unmated females and GIII of 28 uninfected mated females. Animals of group I were mated 4 weeks after infection and half of them were submitted to cesarean section on day 15 after successful mating; the other half was maintained and submitted to cesarean section and sacrificed 14 weeks after infection. Half of the animals of group II were sacrificed seven weeks and the other half 14 weeks after infection. Uninfected animals of group III were treated the same as the animals of group I. The animals were infected with strain 18 of P. brasiliensis by the intracardiac route. We evaluated the disease by the volume of granulomas in different organs, number of fungi in liver and spleen and the immunologic responses [ELISA, Double Immunodifusion (DID), Delayed Hypersensitivity Skin Test (DHT) and Macrophage Migration Inhibition (MMI)]. We studied the infection through the gestation by evaluation of the abortions, morphologic and clinic examinations of the fetuses. Our results showed that the infection did not transfer to the fetus through the placenta, but the number of abortions was larger among infected females. The newborns of GI females were smaller, weighed less and showed little vitality. The disease was more severe and disseminated in infected mated females, especially in the second sacrifice 14 weeks after inoculation, when the total volume of granulomas in them (56.3 mm) was much greater than in the infected unmated females (12 mm).

Annexin-A1 gene expression during liver development and post-translation modification after experimental endotoxemia

Objective and design: We have previously reported a role for annexin-A1 in liver proliferation and tumorogenicity as well as its action as an acute phase protein in a model of endotoxemia in interleukin-6 null mice.Material and methods: In this study, we have investigated the analysis of the gene and protein expression in annexin-A1 null mice and the wild type livers during foetal and adult life, and in the presence of a proinflammatory stimulus.Results: The data indicate a link between the expression of the annexin-A1 as serine-phosphorylated-protein during early events of the inflammatory response and as tyrosine-phosphorylated-form at later time-points, during the resolution of inflammation.Conclusions: The study of annexin-A1 post-translation modification may promote a new annexin-A1 peptide discovery programme to treat specific pathologies.

Sensitive Affimer and Antibody Based Impedimetric Label-Free Assays for C-Reactive Protein

C-reactive protein (CRP) is an acute phase protein whose levels are increased in many disorders. Levels greater than 3 mu g/mL serum have hitherto been considered to indicate pathology, but there is increasing interest in assessments between 0.1 and 10 mu g/mL, which have been found to correlate with severity of risk for cardiovascular disease. We report herein the generation of both antibody and Affimer based impedance immunoassays for CRP that are substantially more sensitive than clinically utilized immunonephelometry and immunoturbidity assessments. Significant in this study is not only the use of a constrained peptide to detect a clinically important target but also that derived electrochemical impedance assays can be highly sensitive even with probes whose relatively weak (mu M) affinities are not amenable to target detection by surface plasmon resonance (SPR). Key to this finding is acknowledging that receptive surfaces of comparatively low initial steric bulk and charge transfer resistance are especially primed to be highly responsive to target binding in electroanalytical assays of this type.

Evolução neurológica e eletrencefalográfica em crianças após meningencefalites purulentas

Twenty children with diagnosed meningitis were available for prospective study; each was submitted to neurological and electroencephalographic examination, Distractability Quotient (Gesell) and Intelligence Quotient (Raven) tests. Patients were followed from 6 months to 3 years after the acute phase of the disease. There is a statistically significant difference between the D.Q of post-meningitic children and the D.Q. of non meningitic controls of the same social class and ages, when the onset of illness was before 30 months of age. No statistically significant correlation was found between the D.Q. and the patient's length of hospitalization or the first cerebrospinal fluid protein level. There is a possibility that significant correlation between the D.Q. and age at onset of illness may be observed by studying a larger number of patients. No statistically significant difference was found between the I.Q. of post-meningitic children and controls when the onset of illness was after age 4.

Indices de atividade da doença inflamatória do intestino: indicadores clínicos e laboratoriais.

The quantification of the degree of activity of inflammatory bowel disease is assuming growing importance nowadays. The activity index of the disease can be attained by clinical and laboratorial indicators. For ulcerative colitis the mostly used clinical parameters are daily bowel movements and presence of bloody diarrhea whereas albumin, hemoglobin, ESR and positive acute phase protein measurements are the laboratory parameters. For Crohn's disease activity besides the daily bowel movements the presence of abdominal pain and discomfort sensation are also frequently used whereas the C-reactive protein is the most used laboratory test which is able to detect the disease reactivation even before the appearance of any clinical sign. The combinations of clinical signs with the laboratory tests earned the sympathy of the specialists and the set of ensembled indicators has been recognized by the author's name. In this sense, the classification of the ulcerative colitis activity originally proposed by Truelove and Witts deserves presently a wide acceptance whereas such agreement is still lacking for Crohn's disease activity. In the mean time, the Bristol index is clinically the most feasible, once the Crohn's disease activity index and the Van Hees index are considered too complex. However the latter indexes are still useful mainly for comparisons among multicentric data. It seems that the currently existing clinical signs used for Crohn's disease activity would be quantitatively improved by adding some easily made laboratory tests such as C-reactive protein.

Role of different proteolytic pathways in degradation of muscle protein from streptozotocin-diabetic rats

In vitro rates of overall proteolysis and the activities of four different proteolytic pathways (lysosomal, Ca2+ dependent, ATP dependent, and ATP independent), as well as rates of protein synthesis, were measured in soleus and extensor digitorum longus (EDL) muscles from streptozotocin- diabetic rats. In the acute phase (1-3 days) of diabetes, there was an increase in overall proteolysis that coincided with an increased activity of the Ca2+-dependent pathway in both soleus and EDL and of the ATP-dependent pathway in EDL. After longer periods (5-10 days) of diabetes, the overall rate of protein degradation decreased and reached values similar to or even lower than those of controls as a result of a reduction in the activities of Ca2+-dependent and ATP-dependent pathways. No change was detected at any time interval in the activity of the intralysosomal proteolytic system in muscles from diabetic animals. Rates of protein synthesis were already reduced 24 h after diabetes induction and decreased further thereafter. Insulin treatment restored to normal the activities of the proteolytic pathways and rates of protein synthesis.

Assessment of vitamin A status in chronic obstructive pulmonary disease patients and healthy smokers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Murine alpha-2-macroglobulin increase during inflammatory responses and tumor growth

Objective and Design: To determine the alpha-2-macroglobulin (alpha2M) levels in mice during acute and chronic inflammatory responses. Materials and Methods: Inflammation was induced by one of the following stimuli: carrageenin, zymosan, lipopolysacharide, thioglycollate, bacilli Calmette Guerin, PPD (in pre-immunized and non-immunized animals) and tumor cells. The concentration of alpha2M was determined in plasma or peritoneal liquid by electroimmunoassay. Results: In all the treatments employed, the plasma levels of alpha2M were higher than in untreated animals. This increase varied from 9%, 24 h after injection up a maximum of 66% 72 h post-injection. When compared to animals injected only with saline, the increases were significant 48 h after treatment with either zymosan or LPS, and 72 h after treatment with either thioglycollate or carrageenin. Treatment with BCG triggers an increase in alpha2M levels after 24 h (18.60%) and 48 h (27.90%). Immunized mice presented higher levels of this protein than non-immunized animals after challenge with PPD. The growth of Ehrlich tumor cells in the peritoneal cavity was directly correlated with the local levels of alpha2M which increased 3.5 fold, 10 days after injection. Conclusions: These results strongly indicate that in mice, the concentration of alpha2M can increase during acute and chronic inflammatory reactions with kinetics dependent on the particular kind of inflammatory agent.

Voltammetric behavior of nitrofurazone and its hydroxymethyl prodrug with potential anti-chagas activity

Chagas' disease is a serious health problem for Latin America. The situation is worsened by the lack of efficient chemotherapy. The two available commercial drugs, benznidazole and nifurtimox, are more effective in the acute phase of the disease. Nitrofurazone is active against Trypanosoma cruzi, however its high toxicity precludes its current use in parasitosis. Hydroxymethylnitrofurazone is a prodrug of nitrofurazone. It is more active against Trypanosoma cruzi than nitrofurazone, besides being less toxic. This work shows the voltammetric behavior of nitrofurazone and a comparison with those of metronidazole and chloramphenicol using cyclic, linear sweep and differential pulse voltammetries. For these drugs also the prediction of the diffusion coefficients using Wilke-Chang equation was performed. The reduction of nitrofurazone is pH-dependent and in acidic medium the hydroxylamine derivative, involving four electrons, is the principal product formed. In aqueous-alkaline medium and with a glassy carbon electrode pre-treatment the reduction of nitrofurazone occurs in two steps, the first involving one electron to form the nitro-radical anion and the second corresponding to the hydroxylamine derivative formation. Hydroxymethylnitrofurazone presented the same voltammetric behavior and electroactivity, indicating that the molecular modification performed in nitrofurazone did not change its capacity to be reduced. A brief discussion regarding the differences in biological activity between the two compounds is also presented. ©2005 Sociedade Brasileira de Química.

Abordagem diagnóstica dos pacientes com suspeita de trombose venosa profunda dos membros inferiores

Deep venous thrombosis is a relatively common disease, which can present pulmonary embolism as a complication in its acute phase, and later the post-thrombotic syndrome. Thus, diagnosis should be made as soon as possible, in order to prevent or minimize such complications. Several studies have shown that the symptoms and the clinical signs are inaccurate for the deep venous thrombosis diagnosis and that complementary exams are necessary. As an attempt to simplify the patients' assessment, Well et al., in 1997, developed a clinical prediction index that combines symptoms, signs and risk factors for deep venous thrombosis and managed to make a simpler approach through an association of this index with the complementary exams. Phlebography has been considered the gold standard of complementary exams. However, since it is an invasive exam and thus subject to complications, other diagnostic methods were introduced aiming at making the diagnostic approach simpler and less invasive. Doppler ultrasound, duplex scan, impedance plethysmography, computed tomography, and blood tests such as the D-dimer are some of the available methods for assessing the patient with suspicion of deep venous thrombosis. Among them, duplex scan has shown excellent accuracy and it is currently widely accepted as the first choice test for approaching the patient with deep venous thrombosis. Several authors have suggested an association of diagnostic methods to simplify and make the assessment of such patients more cost-effective, leading to the introduction of a wide range of diagnostic strategies. The different diagnostic methods used for assessing deep venous thrombosis are discussed, as well as a review of the literature on the accuracy, advantages and disadvantages of these methods. Copyright © 2005 by Sociedade Brasileira de Angiologia e Cirurgia Vascular.

DNA damage and nitric oxide synthesis in experimentally infected Balb/c mice with Trypanosoma cruzi

This study aimed to evaluate whether experimental Chagas disease in acute phase under benznidazole therapy can cause DNA damage in peripheral blood, liver, heart, and spleen cells or induce nitric oxide synthesis in spleen cells. Twenty Balb/c mice were distributed into four groups: control (non-infected animals); Trypanosoma cruzi infected; T. cruzi infected and submitted to benznidazole therapy; and only treated with benznidazole. The results obtained with the single cell gel (comet) assay showed that T. cruzi was able induce DNA damage in heart cells of both benznidazole treated or untreated infected mice. Similarly, T. cruzi infected animals showed an increase of DNA lesions in spleen cells. Regarding nitric oxide synthesis, statistically significant differences (p < 0.05) were observed in all experimental groups compared to negative control, the strongest effect observed in the T. cruzi infected group. Taken together, these results indicate that T. cruzi may increase the level of DNA damage in mice heart and spleen cells. Probably, nitric oxide plays an important role in DNA damaging whereas benznidazole was able to minimize induced T. cruzi genotoxic effects in spleen cells. © 2006 Elsevier Inc. All rights reserved.