PP65 antigenemia in the diagnosis of cytomegalovirus infection in AIDS patients

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

Different nutritional-state indicators of HIV-positive individuals undergoing antiretroviral therapy

This study aimed at learning about the nutritional profile of HIV-positive individuals undergoing antiretroviral therapy and at comparing the performance of nutritional-state indicators. A transversal study was performed on 94 patients attending the Tropical Diseases Outpatient Hospital of Botucatu Medical School (FMB)-UNESP. The body mass index (BMI) and the classification by Papini-Berto (PB) were used to evaluate nutritional state, aiming at detecting malnutrition and obesity. The waist-to-hips ratio (W/HR) and waist circumference (WC) were adopted for identification of abdominal obesity and lipodystrophy. According to BMI, most of the individuals were eutrophic, followed by 30.9% overweight and 6.4% malnourished. By using the PB classification, the frequency of malnourished increased (22.3%). The analysis of the PB classification in relation to BMI indicated that the former presented high sensitivity and good specificity for malnutrition diagnosis, namely 100% and 83%, respectively. The prevalence of abdominal obesity was 7.44% according to WC, and a higher prevalence (38.3%) was observed when taking W/HR into account. There was significant positive association between nutritional diagnosis according to PB and T CD4(+) lymphocyte. The results support the use of PB classification for malnutrition detection as well as that of BMI and W/HR for overweight and fat redistribution.

Assessment of genetic relationship between Klebsiella pneumoniae and Klebsiella oxytoca samples isolated from a dental office

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Production of prostaglandins and leukotrienes by Paracoccidioides brasiliensis

Paracoccidioides brasiliensis is the causative agent of paracoccidioidomycosis, the most prevalent deep mycosis in Latin America. Production of eicosanoids, including prostaglandins and leukotrienes, during fungal infections is theorized to play a critical role on fungal survival and/or growth as well as on host immune response modulation. Host cells are one source of these mediators; however another potential source may be the fungus itself. The purpose of our study was to assess whether P. brasiliensis strains with different degree of virulence (Pb18, Pb265, PbBT79, Pb192) produce both, prostaglandin E(2) (PGE(2)) and leukotriene B(4) (LTB(4)). Moreover, we asked if P. brasiliensis can use exogenous sources of arachidonic acid (AA), as well as metabolic pathways dependent on cyclooxygenase (COX) and lipoxygenase (5-LO) enzymes, for PGE(2) and LTB(4) production, respectively. Finally, a possible association between these eicosanoids and fungus viability was assessed. We demonstrated, using ELISA assays, that all P. brasiliensis strains, independently of their virulence, produce high PGE(2) and LTB(4) levels after a 4-hour culture, which were reduced after 8 hours. However, in both culture times, higher eicosanoids levels were detected when culture medium was supplemented with exogenous AA. Differently, treatment with indomethacin, a COX inhibitor, or MK886, a 5-LO inhibitor, induces a reduction on PGE(2) and LTB(4) levels, respectively, as well as in fungus viability. The data provide evidence that P. brasiliensis is able to metabolize either endogenous or exogenous AA by pathways that depend on COX and 5-LO enzymes for producing, respectively, PGE(2) and LTB(4) that are critical for its viability.

Rotavirus gastroenteritis in a children's hospital specialized in craniofacial malformations

This study aimed to verify the relationship between acute diarrhea provoked by rotavirus and different indicators of craniofacial malformations. In the Hospital for Rehabilitation of Craniofacial Anomalies, 8,724 children with cleft lip and cleft palate were divided into the following groups: acute diarrhea and infection due to rotavirus (C1, n = 62), acute diarrhea (C2, n = 153) and without acute diarrhea (C3, n = 8,509). In C1, 29.03% of the cases consisted of hospital infections associated with the hospitalization period while 38.71% of the patients were aged less than six months. The percentage of children not having breastfed was significantly higher in acute diarrhea groups. Additionally, there was a seasonal prevalence of rotavirus infection between May and October. Finally, the present findings indicate that rotavirus is a predominant etiological agent for gastroenteritis in children with craniofacial malformations. Moreover, among infants younger than six months of age, type of craniofacial malformation, breastfeeding difficulty, socioeconomic level and longer hospitalization period appear to contribute to higher infection morbidity.

Pulmonary tuberculosis: Hematology, serum biochemistry and the relation with the disease duration

The aim of the present study was to analyze the relationship between hematological and biochemical parameters and tuberculosis process activity time according to clinical complaint duration. It was a retrospective study analyzing medical records from 80 pulmonary tuberculosis patients at Botucatu Medical School University Hospital ( Botucatu, São Paulo State, Brazil), who were divided into 2 groups according to clinical complaint duration: Group 1 ( G1) - up to three months; Group 2 ( G2) - over three months. Parameters included: age, gender, bacilloscopy, erythrocyte sedimentation rate ( ESR), platelet count, alpha1-globulin, alpha2-globulin, gamma globulin, mucoprotein, alpha1-acid glycoprotein values, and the presence of risk factors such as smoking, alcoholism, drug addiction, sexual promiscuity, contact with tuberculosis carriers, and previous treatment. Groups were compared by calculating t and p, and Chi-square (X-2) and p. Comparisons revealed a tendency towards smoking with a higher frequency of smokers in G1 ( 0.05< p< 0.10). G1 also tended to present greater platelet values than G2 ( 0.05< p< 0.10) and presented significantly higher ESR values than G2 ( p< 0.05). Other factors did not show any significantly different behavior between groups ( p> 0.05). A correlation was found between ESR, platelet count, smoking and less than three months clinical duration.

Canine trypanosomiasis: etiology of infection and implications for public health

Canine trypanosomiasis, caused by protozoans of the genus Trypanosoma, is divided into two primary types: the American form (Chagas disease), due to Trypanosoma cruzi infection, and the African form (sleeping sickness or surra), provoked by Trypanosoma evansi. This disease was originally enzootic and affected only wild animals, including mammals and birds, which served as reservoirs. Later, it spread to domestic animals such as horses, cattle and dogs. The disease became a zoonosis when contact between rural inhabitants and natural Trypanosoma foci occurred, due to ecological imbalances and increasing migration. Dogs are significantly involved in this context, because they are the main domestic animals and participate in the transmission and maintenance cycles of these parasites. This article reports etiological, epidemiological and public health aspects of canine trypanosomiasis, and the most important peculiarities of this zoonosis in dogs.

Intestinal parasitic infections in HIV/AIDS patients: epidemiological, nutritional and immunological aspects

This study applied a socioeconomic questionnaire designed to evaluate the frequency of intestinal parasites and characterize epidemiological, nutritional, and immunological variables in 105 HIV/AIDS patients - with and without parasitic infections, attending the Day Hospital in Botucatu, UNESP, from 2007 to 2008. Body mass index was calculated and the following tests performed: parasitological stool examinations; eosinophil, IgE, CD4(+) T and CD8(+) T lymphocyte cell counts; albumin test; viral load measure; and TNF-alpha, IFN-gamma, IL-2, IL-5 and IL-10 cytokine levels. Results were positive for parasitic intestinal infections in 12.4% of individuals. Most patients had good socioeconomic conditions with basic sanitation, urban dwellings, treated water supply and sewage, good nutritional and immunological status and were undergoing HAART. Parasites were found at the following frequencies: Entamoeba - five patients (38.5%), Giardia lamblia-four (30.7%), Blastocystis hominis-three (23.0%), Endolimax nana-two (15.4%), and Ascaris lumbricoides - one (7.7%). There were no significant differences between the two groups for eosinophils, albumin, IgE, CD4(+) T and CD8(+) T lymphocytes, INF-gamma, IL-2, or IL-10. Most patients also showed undetectable viral load levels. Significant differences were found for TNF-alpha and IL-5. These results show the importance of new studies on immunodeficient individuals to increase understanding of such variables.

Effect of soluble fiber on hypertriglyceridemia and immune profile in HIV-positive individuals undergoing highly active antiretroviral therapy

The advent of highly active antiretroviral therapy (HAART), since 1996, represented a profound impact on the natural history of HIV-infection by promoting important and sustainable viral replication suppression and increasing survival and quality of life among seropositive patients. Nonetheless, antiretroviral therapy has been observed to be accompanied by metabolic alterations such as dyslipidemia, especially hypertriglyceridemia, insulin resistance, hyperglycemia and lipodystrophy (body fat redistribution). Epidemiological studies have demonstrated a correlation between high triglyceride (TG) levels and higher incidence of coronary artery disease (CAD). Some investigators suggest dietary intervention as part of hyperlipidemia treatment, including an increase in soluble fiber intake (10-25g/day). Whereas some studies have demonstrated that both cholesterol and serum triglyceride levels decrease with the use of food fiber, others have shown just a serum triglyceride decrease, and others failed to observe any alteration in lipid metabolism. The purpose of this study was to assess the effect of soluble fiber (R) (partially hydrolyzed guar gum) supplementation on hypertriglyceridemia and immune profile in HIVpositive individuals on HAART. Nineteen HIV-positive individuals with hypertriglyceridemia (serum levels >= 150 to < 500mg/dl) were studied. of these individuals, 63.16% were males and 36.84% females, with mean age of 43.52 +/- 9.22 years. These individuals had been on the same HAART regimen for at least six months, had no change in therapy during the study and received 20g/day of soluble fiber for four months at pre-established times. Clinical-nutritional, biochemical (total proteins, albumin, globulin, total cholesterol, LDL-c, HDL-c, TG, TG/HDL-c and LDLc/HDL-c), hematimetric (hemoglobin, hematocrit and total lymphocytes), and immunologic (lymphocytes T CD4(+), T CD8(+); T CD4(+)/CD8(+) ratio, viral load, TNF-alpha and IL-6) parameters were assessed in all patients at three time points (M0: pretreatment, M1: 30 days, and M2: four months after intervention). Significance level was set at 5% for all data statistically analyzed. Serum TG and TG/HDL-c ratio reduction was observed at all time points, but statistical significance was found just at M0 and M2. The remaining biochemical, hematimetric and immunologic parameters (lymphocytes T CD4(+), T CD8(+); T CD4(+)/ CD8(+) ratio, and viral load) showed no significant difference at all times. Regarding serum cytokines, TNF-alpha and IL-6 significantly decreased between M0 and M2, and only IL-6 reduced between M1 and M2. The data collected show that dietary and anthropometric parameters remained unchanged excluding potential confounding factors related with the effect of fiber supplementation on serum TG, TNF-alpha and IL-6. Thus, soluble fiber (R) contributed to an important reduction in hypertriglyceridemia and in the serum levels of the proinflammatory cytokines TNF-alpha and IL-6 in HIV-seropositive individuals on HAART. In addition, soluble fiber (R) might have minimized the process of atherosclerosis in these individuals, given that elevated serum levels of TG, TNF-alpha and IL-6 have been associated with the development of these lesions.

Organ tropism during the acute and chronic phases of Trypanosoma cruzi infection in BALB/c mice

The aim of the present study was to investigate the presence of Trypanosoma cruzi in the heart, liver, lung, and kidneys, using hemoculture and PCR analysis, of mice infected with different parasite strains during the acute and chronic phases of infection. Parasitemia curves revealed strain-specific biological behaviors. For the Y and JLP strains, the acute phase of infection started at days six and ten post-infection, parasitemia peaked at days seven and 15 post-infection, the chronic phase started at days nine and 28 post-infection, and animals started dying at days 19 and 120 post-infection, respectively. When the two strains were compared, the JLP strain exhibited reduced and slower replication rates associated with a delayed peak of parasitism and reduced parasite burdens. However, parasites were detected in all studied organs using PCR analysis. The capacity of both strains to infect different organs likely influences disease pathogenesis.

Serological survey for rabies in serum samples from vampire bats (Desmodus rotundus) in Botucatu region, SP, Brazil

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

Leptospira spp. and Toxoplasma gondii antibodies in vampire bats (Desmodus rotundus) in Botucatu region, SP, Brazil

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

Cytokines and acute-phase serum proteins as markers of inflammatory regression during pulmonary tuberculosis treatment

Tuberculosis is still increasing and was declared a worldwide sanitary emergency by the World Health Organization (WHO) in 1995. Its control is difficult due to long treatment duration and lack of markers of treatment success or failure. Cytokines such as IFN-gamma and TNF-alpha, a central factor in immune response against Mycobacterium tuberculosis, are responsible for the interaction between T lymphocytes and the infected macrophage and are also produced during this interaction. As proinflammatory cytokines have a close relationship with mycobacteria clearance, in fact even preceding it, they could be used as markers for inflammatory activity and response to treatment. Proinflammatory cytokines act in the liver and stimulate a strong local and systemic acute-phase response as a result of homeostatic and physiological responses also induced by them. Acute-phase proteins produced by cytokine activity are useful diagnostic markers that could also be used to monitor treatment response as they can be serially quantified. The objective of this study was to evaluate IFN-gamma, TNF-alpha, IL-10 and TGF-beta production in supernatant of peripheral blood mononuclear cell (PBMC) and monocyte (MO) cultures, as well as serum acute-phase response through total protein, albumin, globulin, C-reactive protein (CRP), alpha-1-acid glycoprotein (AGP), and erythrocyte sedimentation rate (ESR) as regression markers of inflammatory response during pulmonary tuberculosis treatment. Twenty blood donors (G1) from the Blood Bank at Botucatu School of Medicine's University Hospital (BSM-UH) were evaluated once and 28 pulmonary tuberculosis patients (G2): 13 from BSM-UH and 15 from the Bauru State Health Secretariat. Patients were evaluated at three moments of treatment: before (M1), at three months (M2), and at the end (M3). Cytokines were determined in 20ml of peripheral blood (ELISA), with or without activation: lipopolysaccharide (LPS) for MO culture and phytohemagglutinin (PHA) for PBMC culture. Acute-phase protein behavior in G2 throughout treatment was: Globulins: M1> M2, M1> M3 (rho < 0.001); CRP: M1> M2> M3 (.< 0.001); AGP for men: M1> M2, M1> M3 (rho < 0.001); ESR for men: M1> M2, M1> M3 (rho < 0.0016) and for women: M1> M2 (.< 0.025). Comparison between cytokine levels found in supernatant of MO and PBMC cultures, with and without stimulus, in G1 and G2 during treatment showed: TNF-alpha (with/ without LPS) at M1: G2> G1; at M2: G2> G1 (rho < 0.001); (without LPS) at M3: G2> G1 (rho < 0.001), (with LPS) at M3: G2> G1 (rho < 0.028); IFN-. (with and without PHA) at M1: G2> G1; at M2: G2> G1 (rho < 0.001); IL-10 (with and without LPS) at M1: G2> G1; at M2: G2> G1; at M3: G2> G1 (rho < 0.001); TGF-beta (with and without LPS) at M1: G2> G1; at M2: G2> G1 (rho < 0.001), (without LPS) at M3: G2> G1 (rho < 0.001). In G2, all cytokines in supernatant of MO and PBMC cultures, with and without stimulus, showed: M1> M2> M3 (rho < 0.01). Levels of globulins, CRP, AGP, and ESR in patients with pulmonary tuberculosis before treatment (M1) were significantly higher than reference values, suggesting their use as diagnostic markers and indicators of treatment. The CRP decreasing values along treatment could be taken as a marker of the regression of inflammatory process and of response to treatment in patients with pulmonary tuberculosis.Regarding cytokines, there was significant increase in TNF-alpha, IFN-gamma, IL-10, and TGF-alpha levels before and at three months treatment, with and without stimulus; in TNF-a and IL-10 lvels, with and without stimulus, as well as in TGF-alpha levels without stimulus at six months. Patients had higher levels of all studied cytokines than controls before treatment, and these values decreased along treatment. In this study, pulmonary tuberculosis patients showed a Th0 cytokine profile before treatment, with the production of both Th1 (IFN-gamma) and Th2 (IL-10) cytokines, in addition to TNF-alpha inflammatory and TGF-alpha regulatory and fibrosis-inducer cytokines. At the end of treatment, all had evolved to Th2 profile, probably in an attempt to reduce the harmful effects of the proinflammatory activity of the Th1 cytokine profile and of the still above-normal levels of TNF-alpha. The high levels of TGF-alpha, also found in these patients, are related to its important role in the extracellular matrix deposition and fibrosis induction that characterize tuberculosis healing process. IFN-gamma was the only cytokine reaching normal levels at the end of treatment, which suggests its use as a marker of response to treatment.

Young ovine death during hyperimmunization: crotalic envenomation or copper toxicosis?

The unfavorable evolution of a young ovine during hyperimmunization process with Crotalus durissus terrificus venom was investigated in order to differentiate its origin between ophidic envenomation and copper toxicosis. Clinical, laboratory, necroscopic and histological exams as well as evaluation and measurement of heavy metals (copper) in the kidneys and in the liver were carried out. Blood counts revealed anemia and serological tests showed high levels of blood urea nitrogen, creatinine, aspartate aminotransferase, creatine phosphokinase, total bilirubin and indirect bilirubin; which indicates liver, kidney and skeletal muscle damages. At necropsy, the animal presented hepatopathy and nephropathy. Histological examination revealed renal and hepatic features that may imply copper intoxication. Copper levels were 237.8 mu g/g in the liver and 51.2 mu g/g in the kidneys. Although the amount of metal found in both organs was below the level that can cause death, according to the literature, anatomopathological signs were suggestive of copper intoxication. Therefore, the hypothesis of metal toxicosis during the hyperimmunization process became more consistent than the crotalic envenomation one.