Oral administration of Hsp65-producing Lactococcus lactis attenuates allergic asthma in a murine model.

AIMS: Allergic asthma is a chronic inflammatory lung disease characterized by a Th2-type immune response pattern. The development of nonspecific immunotherapy is one of the primary goals for the control of this disease. METHODS AND RESULTS: In this study, we evaluated the therapeutic effects of Lactococcus lactis-producing mycobacterial heat shock protein 65 (LLHsp65) in an ovalbumin (OVA)-induced allergic asthma model. OVA-challenged BALB/c mice were orally administrated with LLHsp65 for 10 consecutive days. The results demonstrate that LLhsp65 attenuates critical features of allergic inflammation, like airway hyperresponsiveness and mucus production. Likewise, the treatment decreases the pulmonary eosinophilia and the serum level of OVA-specific IgE. In addition to deviating immune responses towards Th1-cytokine profile, increase regulatory T cells, and cytokine levels, such as IL-6 and IL-10. CONCLUSIONS: Our results reveal that the mucosal immunotherapy of LLHsp65 significantly reduces the overall burden of airway allergic inflammation, suggesting a promising therapeutic strategy for allergic asthma treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: This research reveals new perspectives on nonspecific immunotherapy based on the delivery of recombinant proteins by lactic acid bacteria to treat of allergic disorders.

Establishment of a protocol for the isolation of feline pancreatic islets.

Diabetes mellitus is a common endocrinopathy in cats that is associated with pancreatic islets lesions. Research on isolated islets contributed to the understanding of the pathophysiology of human diabetes. Therefore, by improving the existing methods of isolation in cats, we aimed at increasing islet yield, purity and viability of feline isolated islets. Islet isolation was accomplished by pancreas perfusion with 80ml of Collagenase type IV through the pancreatic duct at the site of the major papilla. The enzymatic digestion was combined with mechanical disruption and controlled by dithizone staining. Purification was performed by filtration and handpicking. Purified islets were plated on extracellular matrix pre-coated plates and cultured for 48h. Feline islets with a high degree of viability and purity were isolated and cultured for the first time. Although the percentage of islet free from the acinar tissue relative to the total number of isolated islets was low compared to other species, the suggested protocol represents a promising progress in the procedure of islet isolation in cats.

Catechol-O-methyltransferase activity in erythrocytes from patients with eating disorders.

PURPOSE: Abnormal feeding has been linked to disruptions in brain dopaminergic activity and recent studies have assessed the role of catechol-O-methyltransferase (COMT) in eating disorders. This is the first study to quantify the soluble catechol-O-methyltransferase (S-COMT) activity in erythrocytes from patients with anorexia nervosa (AN), bulimia nervosa (BN) and binge-eating disorder (BED) and the first study at all to evaluate the COMT on patients with BED. METHODS: Forty blood samples from patients with AN, BN and BED and healthy controls were drawn to evaluate S-COMT activity in erythrocytes by high-performance liquid chromatography and mass spectrometry. Since several patients were being treated with fluoxetine 20 mg, they were included in a different group (BN MED and BED MED). Liver homogenates from rats were used to evaluate baseline S-COMT activity in the presence of fluoxetine by the same in vitro procedures and assays. RESULTS: Erythrocyte S-COMT activity (pmol/mg prt/h) was significantly increased in patients with BN and BED (41.3 ± 6.8 and 41.4 ± 14, respectively) compared to control group (25.3 ± 9.7). In fluoxetine-treated patients with BN, S-COMT activity (15.9 ± 8.8) was decreased compared to the other BN group; however, in BED group, the difference between BED MED and BED was not observed. In patients with AN, no significant difference was found compared to controls. CONCLUSION: Patients with BN and BED presented higher S-COMT activity in erythrocytes, which is in agreement with previous studies on the literature addressing the high-activity COMT allele, Val158, as risk factor for eating disorders. Although in fluoxetine-treated patients with BN the activity of S-COMT was similar to the controls, this is not explained by a direct interaction between fluoxetine and S-COMT as verified in in vitro assays.

Metal and physico-chemical variations at a hydroelectric reservoir analyzed by Multivariate Analyses and Artificial Neural Networks: environmental management and policy/decision-making tools.

This paper compared and evaluated seasonal variations in physico-chemical parameters and metals at a hydroelectric power station reservoir by applying Multivariate Analyses and Artificial Neural Networks (ANN) statistical techniques. A Factor Analysis was used to reduce the number of variables: the first factor was composed of elements Ca, K, Mg and Na, and the second by Chemical Oxygen Demand. The ANN showed 100% correct classifications in training and validation samples. Physico-chemical analyses showed that water pH values were not statistically different between the dry and rainy seasons, while temperature, conductivity, alkalinity, ammonia and DO were higher in the dry period. TSS, hardness and COD, on the other hand, were higher during the rainy season. The statistical analyses showed that Ca, K, Mg and Na are directly connected to the Chemical Oxygen Demand, which indicates a possibility of their input into the reservoir system by domestic sewage and agricultural run-offs. These statistical applications, thus, are also relevant in cases of environmental management and policy decision-making processes, to identify which factors should be further studied and/or modified to recover degraded or contaminated water bodies.

Ensemble asteroseismology of solar-type stars with the NASA Kepler mission.

In addition to its search for extrasolar planets, the NASA Kepler mission provides exquisite data on stellar oscillations. We report the detections of oscillations in 500 solar-type stars in the Kepler field of view, an ensemble that is large enough to allow statistical studies of intrinsic stellar properties (such as mass, radius, and age) and to test theories of stellar evolution. We find that the distribution of observed masses of these stars shows intriguing differences to predictions from models of synthetic stellar populations in the Galaxy.

Efficacy of DNA-hsp65 vaccination for tuberculosis varies with method of DNA introduction in vivo.

A DNA vaccine codifying the mycobacterial hsp65 can prevent infection with Mycobacterium tuberculosis in a prophylactic setting and also therapeutically reduce the number of bacteria in infected mice. The protective mechanism is thought to be related to Th1-mediated events that result in bacterial killing. To determine the best method of hsp65 introduction for vaccination efficacy against tuberculosis (TB), we evaluated the immunogenicity and protection of DNA-hsp65 administered by gene gun bombardment or intramuscular (i.m.) injection of naked DNA. Immunization by gene gun induced immune response with plasmid doses 100-fold lower than those required for intramuscular immunization. However, in contrast to intramuscular immunization, which was protective in these studies, gene gun immunization did not protect BALB/c mice against challenge infection.

Role of trehalose dimycolate in recruitment of cells and modulation of production of cytokines and NO in tuberculosis.

Mice treated with viable Mycobacterium tuberculosis with no glycolipid trehalose dimycolate (TDM) on the outer cell wall (delipidated M. tuberculosis) by intraperitoneal or intratracheal inoculation presented an intense recruitment of polymorphonuclear cells into the peritoneal cavity and an acute inflammatory reaction in the lungs, respectively. In addition, lung lesions were resolved around the 32nd day after intratracheal inoculation. TDM-loaded biodegradable poly-DL-lactide-coglycolide microspheres as well as TDM-coated charcoal particles induced an intense inflammatory reaction. In addition, high levels of interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha), IL-12, IL-10, gamma interferon (IFN-gamma), and IL-4 production were detected in lung cells, and nitric oxide (NO) production was high in culture supernatants of bronchoalveolar lavage cells. These in vivo data were confirmed by in vitro experiments using peritoneal macrophages cultured in the presence of TDM adsorbed onto coverslips. High levels of IFN-gamma, IL-6, TNF-alpha, IL-12, IL-10, and NO were detected in the culture supernatants. Our results suggest that TDM contributes to persistence of infection through production of cytokines, which are important for the recruitment of inflammatory cells and maintenance of a granulomatous reaction. In addition, our findings are important for a better understanding of the immunostimulatory activity of TDM and its possible use as an adjuvant in experiments using DNA vaccine or gene therapy against tuberculosis.

Comparison of different delivery systems of vaccination for the induction of protection against tuberculosis in mice.

The way to deliver antigens and cellular requirements for long-lasting protection against tuberculosis are not known. Immunizations with mycobacterial 65 kDa heat shock protein (hsp65) expressed from J774-hsp65 cells (antigen-presenting cells that endogenously produce hsp65 antigen) or from plasmid DNA, or with the protein entrapped in cationic liposomes, can each give protective immunity similar to that obtained from live Bacillus Calmette Guérin (BCG), whereas injecting the protein in Freund's incomplete adjuvant (FIA) has minimal effect. Protective procedures elicited high frequencies of antigen-reactive alphabeta T cells with CD4+/CD8- and CD8+/CD4- phenotypes. Protection correlated with the abundance of hsp65-dependent cytotoxic CD8+/CD4-/CD44hi cells. The frequency of these cells and the level of protection declined during 8 months after J774-hsp65 or liposome-mediated immunization with hsp65 protein but were sustained or steadily increased over this period after hsp65-DNA or BCG immunizations. IFN-gamma predominated over IL-4 among the hsp65-reactive CD8+/CD4- and CD4+/CD8- populations after J774-hsp65-, hsp65-liposome-, and hsp65-DNA-mediated immunizations, but similar levels of these cytokines prevailed after BCG vaccination.

[Changes in caries prevalence in 7-12-year-old children from Araraquara, São Paulo, Brazil: 1989-1995]. / Mudanças na prevalência de cárie em crianças de sete a 12 anos de Araraquara, São Paulo, Brasil: 1989-1995.

The aim of this study was to compare the prevalence of caries in 7-12-year-old children from Araraquara, São Paulo, Brazil, in 1989 and 1995. Systematic random samples were drawn from the population of children enrolled in public schools. The surveys were carried out by trained examiners using the DMFT index and WHO diagnostic criteria. There was an increase in the percentage of children free of caries in the permanent dentition in all age groups (from 29 per cent in 1989 to 51 per cent in 1995). Amongst 12 year olds, the DMFT index was 3.8 in 1989 and 2.6 in 1995. Decreases were observed in the percentages of children classified in the 1-3 DMFT index category (from 40 per cent in 1989 to 31 per cent in 1995) and in the 4-6 DMFT index category (26.6 per cent in 1989 to 16.5 per cent in 1995). There was also a reduction in the percentage of children with a DMFT of 7 or more (4.4 per cent in 1989 to 1.5 in 1995). The WHO/FDI goal for the year 2000 of a mean DMFT index no more than 3 at the age of 12 years was achieved in this population, and on-going efforts should be made to reduce the percentage of children with caries in order to achieve the WHO/FDI goals for the year 2010.

[Analysis of fluoride concentration in mineral waters in the Araraquara region in Brazil]. / Análise da concentração de flúor em águas minerais na região de Araraquara, Brasil.

It has been shown that people of all ages can benefit from the topical and systemic effects of water fluoridation. However, the increase in consumption of bottled water, either to substitute for or supplement consumption of water from public sources, has implications for safe fluoride supplementation. Taking that into consideration, in 1995 we analyzed the fluoride content in 31 commercial brands of mineral water in the region of Araraquara, state of São Paulo, Brazil. Fluoride concentration as determined by our analysis was compared to the concentration of fluoride specified on each label. Only 25% of the products studied listed the fluoride concentration on their labels. In addition, among 31 mineral water brands, 26 listed the date when the chemical analysis to determine chemical composition had been performed. Of these, 20 had not been put through the annual chemical analysis determined by Brazilian law. Based on these results, if the mineral waters tested had been the only source of drinking water, fluoride supplementation would have been necessary in 69% of the samples analyzed. In the case of children up to 6 years of age who use products containing fluoride, such as topical gels, mouthwashes or toothpastes, supplementation should be recommended only if commercially bottled water is the only source of water used, not only for drinking but for cooking as well.

[Pheochromocytoma--apropos a clinical case]. / Feocromocitoma--a propósito de um caso clínico.

A case of a 43-year-old woman with severe sustained hypertension resistant to many antihypertensive drugs, frequent hypertensive crisis and symptoms suggestive of pheochromocytoma (symptomatic triad) is presented. Three of the four determinations of the urinary catecholamines metabolites have been normal as it was the only determination of plasmatic catecholamines. Abdominal sonography and CT scan detected a left adrenal mass, that have been histologically confirmed, after surgery, to be a pheochromocytoma. After adrenalectomy, the patient symptoms disappeared but she maintained mild hypertension easily controlled with drugs. The finding of normal plasmatic and urinary catecholamines values in a patient with sustained hypertension may suggest that we are handling with a case of essential hypertension and a superimposed pheochromocytoma with paroxysmal secretion. Some considerations are made essentially about specificity and sensitivity of diagnostic tests.

[Primary hyperaldosteronism. A report of 2 clinical cases]. / Hiperaldosteronismo primário. Apresentação de dois casos clínicos.

We describe two clinical cases of primary hyperaldosteronism caused by adrenal adenoma (Conn's syndrome) with very different forms of clinical presentation. Some theoretical considerations about diagnosis and treatment of the disease are made.