Immunotherapy against murine experimental visceral leishmaniasis with the FML-vaccine

The fucose mannose ligand (Leishmania donovani FML)-saponin vaccine has earlier shown its immunoprophylactic potential against visceral leishmaniasis in the CB hamster (87.7% of parasite load reduction), Balb/c (84.4%) and Swiss albino mouse (85-93%) models. In this investigation its specific immunotherapeutic efficacy against L. donovani infection in Balb/c mice was studied. The effects of vaccine treatment on the Immoral response, delayed type of hypersensitivity to promastigote lysate (DTH), cytokine levels in sera and reduction of the liver parasitic load of L. donovani infected mice, were examined. The types and subtypes of anti-FML antibodies increased significantly in the vaccinees over the saline and saponin controls. As expected for a saponin vaccine, the highest ratios were found in relation to IgG1, IgG2a and IgG2b (4.4, 5 and 2.5, respectively). The DTH response and the in vitro ganglion cell proliferative response against FML antigen were also significantly higher than controls (P < 0.005). Concomitantly, an impressive and specific decrease of liver parasitic burden was detected only in vaccine-treated animals (94.7%). Our results indicate that the therapeutic FML-vaccine has a potent effect on modulation of the murine infection leading to the reduction of parasitic load and signs of disease, being a new potential tool in the therapy and control of visceral leishmaniasis. (C) 2003 Elsevier Ltd. All rights reserved.

Electrostatic force microscopy as a tool to estimate the number of active potential barriers in dense non-Ohmic polycrystalline SnO2 devices

In the present work, electroactive grain boundaries of highly dense metal oxide SnO2-based polycrystalline varistors were determined by electrostatic force microscopy (EFM). The EFM technique was applied to identify electroactive grain boundaries and thus estimate the amount of active grain boundary, which, in the metal oxide SnO2-based varistor, was calculated at around 85%, i.e., much higher than that found in traditional metal oxide ZnO-based varistors. The mean potential barrier height value obtained from the EFM analysis was in complete agreement with the values calculated from the C-V measurements, together with a complex capacitance plane analysis that validates the methodology proposed here. (c) 2006 American Institute of Physics.

Obesity enhances eosinophilic inflammation in a murine model of allergic asthma

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Characterization of dense lead lanthanum titanate ceramics prepared from powders synthesized by the oxidant peroxo method

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

In vitro Culture of Bovine Embryos in Murine ES Cell Conditioned Media Negatively Affects Expression of Pluripotency-Related Markers OCT4, SOX2 and SSEA1

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

Ketoconazole in the treatment of experimental murine paracoccidioidomycosis

In a murine model of chronic disseminated paracoccidioidomycosis (strain 18; intravenous route), Ketoconazole (200 mg/kg in 0.2% agar) was given daily by gavage in three different schedules. Continuous treatment from an early stage of infection (day 3) up to week 20 was the most effective protocol, leading to remission of histopathological lesions and of both humoral and cellular anti-P. brasiliensis immune response, and clearance of the fungus in lungs; only 1 treated animal at week 20 showed pulmonary granulomas, although less extensive than control mice. Continuous treatment from early stage up to week 8, followed by a 16 week-period of drug discontinuity, caused remission of lesions in all but 3 treated mice which showed active pulmonary paracoccidioidomycosis similar to controls (14.2% of unresponsiveness to treatment). The continuous Ketoconazole protocol since a late stage of infection (week 4) up to week 20 produced a slower remission of lesions and immune response when compared with the first drug schedule. In this model of paracoccidioidomycosis, Ketoconazole showed no detectable side-effects and was a very effective drug especially in a prolonged administration protocol from an early stage of infection.

Sex determination of murine and bovine embryos using cytotoxicity and immunofluorescence assays

Murine and bovine embryos were sexed by using polyclonal and monoclonal anti-H-Y antibodies in cytotoxic and immunofluorescence assays. The specificity of 3 different sources of antibodies are compared. Sexing was accomplished by complement-mediated cytotoxicity or by H-Y antigen detection using a anti-mouse Ig fluorescein conjugate. Cytogenetic ascertainment for correct sex identification yielded a score of about 85% in both species, while the immunofluorescence assay yielded a score of 83%. © 1993.

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.

The adverse effect of a high energy dense diet on cardiac tissue

Purpose: To determine whether a high energy dense diet intake increases oxidative stress and alters antioxidant enzymes in cardiac tissue. Design: A randomized, controlled study. Ninety-day-old female rats were randomly divided into two groups: one fed with a low energy dense diet (LE; 3.0 kcal g-1) and one with a high energy dense diet (HE; 4.5 kcal g-1). Materials and Methods: After 8 weeks of treatment, the animals were fasted overnight and sacrificed by decapitation. The serum was used for glucose, triacylglycerol, cholesterol, low-density lipoprotein (LDL)-cholesterol and high-density lipoprotein (HDL)-cholesterol determinations. The glycogen, lipoperoxide, lipid hydroperoxide, superoxide dismutase, glutathione peroxidase, lactate dehydrogenase, citrate synthase, total and non-protein sulphhydryl groups were determined in cardiac tissue. Results: HE decreased the myocardial glycogen content and increased the lactate dehydrogenase/citrate synthase ratio, indicating an increased glycolytic pathway and a shift from myocardial aerobic metabolism. HE-treated female rats showed increased lipoperoxide and hydroperoxide levels in cardiac tissue. Although no alterations were observed in the total sulphhydryl group and superoxide dismutase activities, glutathione peroxidase and the non-protein sulphhydryl group were significantly decreased in HE-treated animals. Conclusions: Although no alterations were observed in energy intake, HE induced an increased intake of fat and carbohydrate and an increased rate of weight gain. HE intake induced alterations in markers of oxidative stress in cardiac tissue. Hydrogen peroxide is an important toxic intermediate in the development of cardiac oxidative stress by HE. The specific nutrient content, such as fat and carbohydrate, rather than caloric intake, appears to be the main process inducing oxidative stress in HE-treated female rats.

Critical protective role for annexin 1 gene expression in the endotoxemic murine microcirculation

The inflammatory response is a protective process of the body to counteract xenobiotic penetration and injury, although in disease this response can become deregulated. There are endogenous biochemical pathways that operate in the host to keep inflammation under control. Here we demonstrate that the counter-regulator annexin 1 (AnxA1) is critical for controlling experimental endotoxemia. Lipopolysaccharide (LPS) markedly activated the AnxA1 gene in epithelial cells, neutrophils, and peritoneal, mesenteric, and alveolar macrophages-cell types known to function in experimental endotoxemia. Administration of LPS to AnxA1-deficient mice produced a toxic response characterized by organ injury and lethality within 48 hours, a phenotype rescued by exogenous application of low doses of the protein. In the absence of AnxA1, LPS generated a deregulated cellular and cytokine response with a marked degree of leukocyte adhesion in the microcirculation. Analysis of LPS receptor expression in AnxA1-null macrophages indicated an aberrant expression of Toll-like receptor 4. In conclusion, this study has detailed cellular and biochemical alterations associated with AnxA1 gene deletion and highlighted the impact of this protective circuit for the correct functioning of the homeostatic response to sublethal doses of LPS. Copyright © American Society for Investigative Pathology.

Biocompatibility of acellular dermal matrix graft evaluated in culture of murine macrophages

The acellular dermal matrix allograft has been used as an alternative to autogenous palatal mucosal graft. The aim of this study was the evaluation of the biocompatibility of an acellular dermal matrix (AlloDerm®) in culture of macrophages. For hydrogen peroxidase determination we used the method of Pick & Kesari, and the Griess method for nitric oxide determination,. Statistical analysis showed no significant difference (p ≤ 0,05) in the release of nitric oxide and hydrogen peroxide by the macrophages exposed to acellular dermal matrix and the negative control. The results suggest that acellular dermal matrix did not activate the cell inflammatory response.

Cytokine and antibody production during murine leptospirosis

The aim of the present study was to investigate the kinetics of humoral and cellular responses during leptospirosis. We observed that the presence of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) was associated with antibody production and bacterial recovery, and the compromising of both TNF-α and IL-6 in the immunopathogenesis of leptospirosis during an experimental infection of BALB/c mice inoculated with Leptospira interrogans serovar Canicola was verified. Results showed higher levels of TNF-α and IL-6 in the initial phase of infection, in which the greatest bacterial clearance was observed. However, when the bacterial recovery was compared with the kinetics of the production of antibodies, the results revealed a kinetics proportionally inverted to antibody production. This fact may be related to some inhibitory factor which could be responsible for the selective suppression of the cellular immune response. We concluded that during leptospirosis there was a greater mobilization of the cellular immune response activity, mainly in the initial phase of the infectious process, for posterior involvement of the humoral response, and that both TNF-α and IL-6 could be associated with the immunopathogenesis of the disease.

Cytomegalovirus and Toxoplasma gondii seroprevalence in a Brazilian liver transplant waiting list

Cytomegalovirus (CMV) disease is a major cause of morbidity and mortality in solid organ transplantation. Disseminated toxoplasmosis after liver transplantation is a rare but fatal event. Serologic screening of the donor and the recipient is essential to prophylactic management, early diagnosis and therapeutic strategies to minimize the consequences of these infections. The aim of the present study was to determine the seroprevalence of CMV and Toxoplasma gondii (TG) in a Brazilian liver transplant waiting list (LTWL). Serological data were collected from 44 candidates on the LTWL between May 2003 and November 2004. Serological investigation of antibodies IgM and IgG against CMV (anti-CMV) and TG (anti-T. gondii) was performed using fluorometry commercial kits. IgG anti-CMV was positive in 37 patients (94.9%) out of 39 available results. There were not IgM anti-CMV positive results. Out of 36 analyzed patients, 22 (61.1%) presented positive IgG anti-T. gondii and none had positive IgM anti-T. gondii. The high CMV seroprevalence among our LTWL reinforces the need for appropriate protocols to avoid related complications, like reactivation and superinfection by CMV. Environmental and drug prophylactic strategies against primary infection and reactivation, as well as early diagnosis and treatment of toxoplasmosis complications, are essential for the good outcome of transplant patients.