Immunomodulation and protection induced by DNA-hsp65 vaccination in an animal model of arthritis

We described a prophylactic and therapeutic effect of a DNA vaccine encoding the Mycobacterium leprae 65- kDa heat shock protein (DNA-hsp65) in experimental murine tuberculosis. However, high homology of the vaccine to the corresponding mammalian hsp60, together with the CpG motifs in the plasmidial vector, could trigger or exacerbate an autoimmune disease. In the present study, we evaluate the potential of DNA- hsp65 vaccination to induce or modulate arthritis in mice genetically selected for acute inflammatory reaction (AIR), either maximal (AIRmax) or minimal (AIRmin). Mice immunized with DNA-hsp65 or injected with the corresponding DNA vector (DNAv) developed no arthritis, whereas pristane injection resulted in arthritis in 62% of AIRmax mice and 7.3% of AIRmin mice. Administered after pristane, DNA- hsp65 downregulated arthritis induction in AIRmax animals. Levels of interleukin (IL)- 12 were significantly lower in mice receiving pristane plus DNA- hsp65 or DNAv than in mice receiving pristane alone. However, when mice previously injected with pristane were inoculated with DNA- hsp65 or DNAv, the protective effect was significantly correlated with lower IL-6 and IL-12 levels and higher IL-10 levels. Our results strongly suggest that DNA-hsp65 has no arthritogenic potential and is actually protective against experimentally induced arthritis in mice.

EPIZOOTIC BOTULISM OF CATTLE IN BRAZIL

The first diagnosis of botulism in cattle in Brazil and it's epizootiology are reviewed. The high prevalence of the disease raised on phosphorus deficient pastures in Savanna regions has caused severe economic losses in the past.The temperature induced microcomplement fixation test (TIMCF) confirmed the clinical-pathological diagnosis in all of the 24 cases studied by this method.The most important reason why botulism has not been controlled satisfactorily in Brazil is the lack of an available effective vaccine (type C and D). Additional prophylactic measures are phosphorus supplementation and removal of carcasses from the pasture.

Use of topical mitomycin C in myoplasty of the medial rectus muscle of rabbits

To possibly reduce postoperative adhesions that occur after ocular myoplasties, we investigated the topical effects of 0.04% mitomycin C on the repaired areas of the medial rectus muscle using an equine renal capsule preserved in 98% glycerin for reinforcement of the sutures. Twenty-four rabbits, divided into two groups of 12 animals each [untreated (control) and treated group (MMC)], were submitted to surgical rupture of the medial rectus muscle of one eye and repair of the defect 24 h later with sutures and an equine renal capsule. Post-operative prophylactic treatment of the two groups consisted of the administration of eye drops containing neomycin, polymyxin B and dexamethasone at regular 6-h intervals for eight consecutive days and daily rinsing with physiological saline. MMC animals received additional treatment with topical 0.04% mitomycin C every 6 h for 14 consecutive days. Slit lamp biomicroscopy showed greater irritation of the ocular surface in MMC animals during the first days post operatively. Adhesions were observed at 15 and 30 days of assessment in the two groups, but were more extensive in control animals at 60 days. Histopathology revealed inflammatory exudation in both groups, which was greater in MMC animals. Mitomycin C (0.04%) instilled at 6-h intervals for 14 consecutive days reduced the occurrence of fibrosis in the myoplastic areas. However, the equine renal capsule was found to be of little benefit for the reinforcement of myoplasties.

Evaluation of the isoflurane-sparing effects of lidocaine and fentanyl during surgery in dogs

Objective-To evaluate the isoflurane-sparing effects of lidocaine and fentanyl administered by constant rate infusion (CRI) during surgery in dogs.Design-Randomized prospective study.Animals-24 female dogs undergoing unilateral mastectomy because of mammary neoplasia.Procedures-After premedication with acepromazine and morphine and anesthetic induction with ketamine and diazepam, anesthesia in dogs (n = 8/group) was maintained with isoflurane combined with either saline (0.9% NaCl) solution (control), liclocaine (1.5 mg/kg [0.68 mg/lb], IV bolus, followed by 250 mu g/kg/min [113 mu g/lb/min], CRI), or fentanyl (5 mu g/kg [2.27 mu g/lb], IV bolus, followed by 0.5 mu g/kg/min [0.23 mu g/lb/min], CRI). Positive-pressure ventilation was used to maintain eucapnia. An anesthetist unaware of treatment, endtidal isoflurane (ETiso) concentration, and vaporizer concentrations adjusted a nonprecision vaporizer to maintain surgical depth of anesthesia. Cardiopulmonary variables and ETiso values were monitored before and after beginning surgery.Results-Heart rate was lower in the fentanyl group. Mean arterial pressure did not differ among groups after surgery commenced. In the control group, mean +/- SD ETiso values ranged from 1.16 +/- 0.35% to 1.94 +/- 0.96%. Fentanyl significantly reduced isoflurane requirements during surgical stimulation by 54% to 66%, whereas the reduction in ETiso concentration (34% to 44%) observed in the lidocaine group was not significant.Conclusions and Clinical Relevance-Administration of fentanyl resulted in greater isoflurane sparing effect than did liclocaine. However, it appeared that the low heart rate induced by fentanyl may partially offset the improvement in mean arterial pressure that would be expected with reduced isoflurane requirements.

Immune modulation induced by tuberculosis DNA vaccine protects non-obese diabetic mice from diabetes progression

We have described previously the prophylactic and therapeutic effect of a DNA vaccine encoding the Mycobacterium leprae 65 kDa heat shock protein (DNA-HSP65) in experimental murine tuberculosis. However, the high homology of this protein to the corresponding mammalian 60 kDa heat shock protein (Hsp60), together with the CpG motifs in the plasmid vector, could trigger or exacerbate the development of autoimmune diseases. The non-obese diabetic (NOD) mouse develops insulin-dependent diabetes mellitus (IDDM) spontaneously as a consequence of an autoimmune process that leads to destruction of the insulin-producing beta cells of the pancreas. IDDM is characterized by increased T helper 1 (Th1) cell responses toward several autoantigens, including Hsp60, glutamic acid decarboxylase and insulin. In the present study, we evaluated the potential of DNA-HSP65 injection to modulate diabetes in NOD mice. Our results show that DNA-HSP65 or DNA empty vector had no diabetogenic effect and actually protected NOD mice against the development of severe diabetes. However, this effect was more pronounced in DNA-HSP65-injected mice. The protective effect of DNA-HSP65 injection was associated with a clear shift in the cellular infiltration pattern in the pancreas. This change included reduction of CD4(+) and CD8(+) T cells infiltration, appearance of CD25(+) cells influx and an increased staining for interleukin (IL)-10 in the islets. These results show that DNA-HSP65 can protect NOD mice against diabetes and can therefore be considered in the development of new immunotherapeutic strategies.

Immune regulatory effect of pHSP65 DNA therapy in pulmonary tuberculosis: activation of CD8(+) cells, interferon-gamma recovery and reduction of lung injury

A DNA vaccine based on the heat-shock protein 65 Mycobacterium leprae gene (pHSP65) presented a prophylactic and therapeutic effect in an experimental model of tuberculosis. In this paper, we addressed the question of which protective mechanisms are activated in Mycobacterium tuberculosis-infected mice after immune therapy with pHSP65. We evaluated activation of the cellular immune response in the lungs of infected mice 30 days after infection (initiation of immune therapy) and in those of uninfected mice. After 70 days (end of immune therapy), the immune responses of infected untreated mice, infected pHSP65-treated mice and infected pCDNA3-treated mice were also evaluated. Our results show that the most significant effect of pHSP65 was the stimulation of CD8(+) lung cell activation, interferon-gamma recovery and reduction of lung injury. There was also partial restoration of the production of tumour necrosis factor-alpha. Treatment with pcDNA3 vector also induced an immune stimulatory effect. However, only infected pHSP65-treated mice were able to produce significant levels of interferon-gamma and to restrict the growth of bacilli.

Serologic profile in hepatitis B virus and hepatitis C virus in a Brazilian liver transplant waiting list

Chronic viral hepatitis is currently the most common indication for liver transplantation (OLT). Knowing the serological profile of patients on the liver transplant waiting list (LTWL) is essential to manage prophylactic and therapeutic strategies pre- and post-OLT. The aim of this study was to determine the hepatitis B virus (HBV) and hepatitis C virus (HCV) serological profile on the LTWL.Methods. Serological data were collected from 44 candidates included on, the LTWL from May 2003 to November 2004. HBV and HCV serological profiles were performed by microenzyme immunoassay.Results. Twenty-eight patients (66.7%) lacked H13V serological markers. Anti-HBs was detected in 9.5% and was positive for HBsAg, anti-HBc, IgM anti-HBc, or HbeAg in 4.8% of patients, probably related to reactivation of chronic infection. In 7.1% of patients, the markers demonstrated serological cure of infection. In HCV patients, 41.5% were positive. There was H13V and HCV co-infection in 12.2% of patients.Conclusion. HBV infection in 21.4% of the patients corroborates the need to use more efficient protocols for prophylactic and therapeutic management pre- and post-OLT. The high prevalence of HCV infection reinforces the need to follow adequate protocols to avoid related complications and guarantee rational and universal use of more efficient drugs.

Randomized controlled trial on prevention of postcesarean infection using penicillin and cephalothin in Brazil

Background. There is a need to assess the effects of different antibiotic administration models on infectious complications among women from low-income populations who undergo cesarean delivery, and the cost benefit. Design. Randomized, blinded controlled clinical trial study of a single preoperative dose of cephalothin, versus a postcesarean scheme for infection prophylaxis, versus no antibiotics. Methods. The setting was a tertiary Brazilian center with 1,500 deliveries annually. Pregnant women (n = 600) with an indication for emergency or elective cesarean section were randomly allocated consecutively to one of three groups and treated as follows: Group 1 (n = 200), no antibiotics; Group 2 (n = 200), the standard antibiotics scheme followed at this center; Group 3 (n = 200), a single dose of intravenous cephalothin 2 g, intraoperatively. Main outcome measurements. Prevalences of wound infection, puerperal and postcesarean infections, and costs of antibiotics used. Results. Antibiotics reduced the incidence of puerperal infection, but did not change the percentages of wound and postcesarean infections and no use of antibiotics increased the puerperal infection risk sixfold. Cephalothin reduced the relative risk of puerperal infection by 89% (95% confidence interval: 7-87%). Penicillin reduced it by 78%, but this was not statistically significant. No deaths occurred. The costs of the two schemes were similar (almost US$1.00). Conclusions. Prophylactic cephalothin use was associated with decreased postcesarean puerperal infection and presented a cost benefit.

Predictors of incorrectness in antimicrobial prescription in the Bauru State Hospital (Bauru city, São Paulo State, Brazil)

The inappropriate use of antimicrobials in hospitals presents a negative impact on patient outcome and is associated with the emergence and spread of multidrug-resistant microorganisms. Antimicrobial stewardship programs (ASPs) have been instituted in order to improve the quality of prescriptions in hospitals. In this setting, the identification of patterns of inappropriate antimicrobial prescription is a valuable tool that allows ASPs to identify priorities for directing educative/restrictive policies. With this purpose, a study was conducted in the Bauru State Hospital, a teaching hospital with 285 beds affiliated to the Botucatu Medical School, São Paulo State University. The hospital maintains an active ASP since it was opened, in 2002. We selected 25% of the requests for parenteral antimicrobials (RPAs) from 2005 for analysis. Prescriptions for prophylactic purposes were excluded. All other RPAs were classified according to a modified Kunin and Jones categories. Univariate and multivariable analyses were performed to identify predictors of general inappropriateness and of specific prescription errors. Prescriptions classified as "appropriate'' or "probably appropriate" were selected as controls in all stages of the study. Among 963 RPAs included in our study, 34.6% were inappropriate. General predictors of inappropriateness were: prescription on weekends/holidays (OR = 1.67, 95% CI = 1.20-2.28, p = 0.002), patient from intensive care unit (OR = 1.57, 95% CI = 1.11-2.23, p = 0.01), peritoneal (OR = 2.15, 95% CI = 1.27-3.65, p = 0.004) or urinary tract infection (OR = 1.89, 95% CI = 1.25-2.87, p = 0.002), combined therapy with two or more antimicrobials (OR = 1.72, 95% CI = 1.15-2.57, p = 0.008) and prescriptions including penicillin (OR = 2.12, 95% CI = 1.39-3.25, p = 0.001) or first-generation cephalosporins (OR = 1.74, 95% CI = 1.01-3.00, p = 0.048). Previous consultation with an infectious diseases (ID) specialist had a protective effect against inappropriate prescription (OR = 0.34, 95% CI = 0.24-0.50, p < 0.001). Factors independently associated with specific prescription errors varied. However, consultation with an ID specialist was protective against both unnecessary antimicrobial use (OR = 0.04, 95% CI = 0.01-0.26, p = 0.001) and requests for agents with insufficient spectrum (OR = 0.14, 95% CI = 0.03-0.30, p = 0.01). In conclusion, the analysis of factors predictive of inappropriateness in antimicrobial prescription allowed us to identify issues requiring intervention. Also, it provided a positive feedback of the ASP efficacy, demonstrating the importance of previous consultation with an ID specialist to assure the quality of antimicrobial prescriptions.

Investigação de parasitoses e/ou comensais intestinais em manipuladores de alimentos de escolas públicas

The objective of this work was to identify intestinal parasites and/or commensals among food handlers of public schools in the city of Patos of Minas (MG). The fecal samples were analyzed by Lutz's method (1919). The positive results were 21.7% (13 cases) for parasites and/or commensal. The frequence of intestinal parasites and/or commensal of food handlers in the public schools showed low positive, probably as a result of good habits of personal hygiene and prophylactic treatment of parasitic.

Anticoccidial effects of coumestans from Eclipta alba for sustainable control of Eimeria tenella parasitosis in poultry production

The aim of this study was to evaluate the anticoccidial efficacy of a product containing coumestans from Eclipta alba. Experimental conditions were set up as to reproduce the environment conditions for husbandry adopted in commercial broiler farms. Broilers were raised in broiler chicken shed provided with feeders, drinkers, illumination and temperature control systems and floor covering to afford an adequate nourishing environment. Male Cobb broilers (240) were assigned to four experimental groups being each experimental group set apart in rice straw-covered shed isolated with wire mesh. One-day-old broilers were reared in a coccidian-free environment with ad libitum supply of filtered water and freely available standard feed, from the 1st to the 35th day of life. The T1 group received standard feed (negative control); T2 was treated with standard feed supplemented with 66 ppm of salinomycin (positive control); groups T3 and T4 had standard feed supplemented with the ethyl acetate fraction from methanolic extract of E. alba aerial parts, which contains the coumestans WL and DWL (120 and 180 ppm, respectively). The chicken broilers were individually infected with 2 x 104 oocysts of Eimeria tenella when they were 14 days old and were monitored weekly to evaluate zootechnical parameters such as weight gain and food conversion ratio. Counting of coccidial oocyst in chiken feces was assessed from random samples, from the 21st to 28th days of life, which corresponded to 7-14 days after the infection. Five chickens selected at random from each experimental group were subsequently euthanized at 21, 28 or 35 days of life to determine the lesion score in the cecal region and to excise a cecum portion for histopathological evaluation. The group treated with coumestans from E. alba presented an average weight gain and food conversion ratio higher than the negative control group and similar to the mean value of the positive control group. Coumestan-treated groups showed a significant decrease in the oocyst counting since the 21th day of life and displayed a reduced number of macroscopic lesions. Histopathological evaluations of cecum fragments showed that both treatments induced the migration of defense cells at the site of infection. A severe destruction of the cecal lining was found in the intestinal tract of broilers fed with a coumestans dose of 180 ppm. Overall, our results validate the use of a phytotherapy containing E. alba coumestans at a dose of 120 ppm as a therapeutic or prophylactic agent against avian coccidiosis. (C) 2010 Elsevier B.V. All rights reserved.

Temporal IgG subclasses response in dogs following vaccination against Leishmania with Leishmune (R)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Prevalence and risk factors for human toxoplasmosis in a rural community

Toxoplasma gondii infection may lead to important pathological questions, especially in rural areas, where several sources of infection exist. Therefore, it is important to determine risk factors in order to establish adequate prophylactic measures. The present study aimed to assess the prevalence and risk factors involved in human toxoplasmosis infection in a rural community, in Eldorado, Mato Grosso do Sul State, Brazil. This community was composed of 185 farms - with 671 inhabitants - from which 20 were randomly chosen. In these farms, blood samples were collected from rural workers, who also answered a risk factor questionnaire. Serum samples were analyzed by means of direct agglutination test for the detection of anti-Toxoplasma gondii antibodies. From 73 samples collected, 79.45% were positive. None of the studied variables was significantly associated with the prevalence of the infection. However, among the individuals who reported eyesight impairments, 94.4% had anti-T. gondii antibodies, compared with 74.0% who did not report eyesight changes (p = 0.0594). Moreover, most individuals in the study (68.20%) were older than 18 years and presented 84.44% positivity, compared with 66.67% of positive individuals younger than 18 years old. We were able to conclude that a high prevalence of antibodies did not imply significant associations with the risk factors studied.

Ocorrência de mastite bovina causada por Staphylococcus sp., Streptococcus sp. E Candida sp. em uma propriedade rural no município de Indianópolis – Minas Gerais

The objective of this study was to evaluate the occurrence of bovine mastitis by Staphylococcus sp., Streptococcus sp. and Candida sp. in a rural area of Indianopolis, Minas Gerais. It was realized the California Mastitis Test (CMT) in six collect, a total 671 of milk sample positive. Then the microbiological examination was performed, where the results revealed the presence of 137 milk samples with microbial multiplication. These, showed the presence of Staphylococcus aureus (45.2% of strains), other coagulase negative Staphylococcus (10.2%), Staphylococcus epidermidis (9.4%), Staphylococcus simulans (5.8%), other coagulase negative (15.3%), Streptococcus agalactiae (7.2%), other Streptococcus sp. (5.1%) and yeasts (1.4%). It was found that 100% of Staphylococcus were susceptible to rifampicin, gentamicin and ciprofloxacin; but, resistant to penicillin, tetracycline, and oxacillin. Regarding antimicrobial susceptibility Streptococcus, were employed, except to clindamycin, erythromycin and tetracycline. We conclude that there is a great necessity of proper hygiene practices and taking prophylactic measures taken in order to reduce the infection of animals caused by infectious microorganisms and resistance.

Considerações sobre o câncer localmente avançado de mama (CLAM) acrescidas de identificação dos receptores de estrogênios usando anticorpos monoclonais.

The effects of therapy in locally advanced breast cancer submitted to combined conventional telecobalt therapy plus chemotherapy with cyclophosphamide and 5-fluorouracil were studied in 49 patients. Associated to radical mastectomy in operable cases. Local tumor control was achieved in 86.7%. There were no local recurrences in those submitted to surgery but they reached 21.7% in inoperable patients who received only radiation therapy and chemotherapy. The median follow-up time for dead patients was 29.5 months and for living patients 79.3 months. The index of complete responses was 24.5% and the median disease free interval was 22.9 months. The overall survival rate, between three and five years, was 32.7%. Estrogen receptors were identified by using immunohistochemical assay ER-ICA and monoclonal antibody H222-SP gamma, Abbott. There were no differences in the complete response index, disease free interval and survival rates, among ER-positive and ER-negative patients, explained by the far advanced stage of the disease. ER-positivity was significantly correlated with histological features of the tumors: cell differentiation, presence of elastosis, absence of lymphocytic infiltration and absence of tumor necrosis.