Immunization of bovines using a DNA vaccine (pcDNA3.1/MSP1b) prepared from the jaboticabal strain of Anaplasma marginale

Anaplasma is a tick-borne ehrlichial pathogen of cattle that causes the disease, anaplasmosis. In the present study, a total of 11 Anaplasma marginale seronegative calves were assigned into two groups: one immunized (G1, n = 6) and one nonimmunized-control (G2, n = 5). Six calves were immunized by using a DNA vaccine containing the gene of a major surface protein, MSP1b, encoded by the plasmid identified as pcDNA3.1/MSPIb. Calves received three intramuscular inoculations of 100 mug of pcDNA3.1/MSP1b at a 20-day interval. The control group received buffer phosphate at the same schedule as the experimental group. The immune response elicited by immunization with pcDNA3.1/MSP1b was evaluated in mice and calves. Twenty days following initial immunization, specific serum antibody from four BALB/c mice bound MSP1b in inummoblots. Sixty days after the last immunization, all calves were challenged with cryopreserved A. marginale at a dose of 10(4) parasites/mL/animal by intravenous injection. Results of packed cell volume (PCV) and detection of infected erythrocytes in all experimental groups revealed that the decrease of PCV and detection of infected erythrocytes occurred at 28 to 42 days after challenge. Mean temperature values did not increase over 39.85degreesC. Antibodies developed by immunized bovines from G2 were detected 14 days after challenge. MSP1b was characterized during the immunization period and MSP2 was the most predominant polypeptide at the challenge period. DNA of A. marginale was detected in all groups just after challenge by nested PCR assay. It can be concluded that all immunized bovines were partially protected against homologous challenge.

Liver lesions produced by aflatoxins in Rana catesbeiana (bullfrog)

This study describes alterations induced in Rana catesbeiana (bullfrog) liver after extended dietary exposure to aflatoxins (AFs). Bullfrogs of both sexes were fed for 120 days a commercial chow blended with a rice bran-based mixture of Al's containing 667.0, 11.65, 141.74, and 3.53 mg/kg of AFs B1, B2, G1, and G2, respectively. Animals were sacrificed on study days 45, 90, and 120. Severe and progressive liver lesions with structural collapse, increased hepatocyte and biliary duct cell proliferation, appearance of basophilic hepatocytes, and diffuse scarring, were observed at all time points. There were no quantitative alterations in the liver melanomacrophage centers of the AFs-exposed animals. Increased amounts of lipid hydroperoxides, indicative of ongoing oxidative stress, were more evident in the Addutor magnum muscle than in the AFs-damaged livers. No tumors were found in the R. catesbeiana livers after 120 days of exposure to relatively high doses of AFs. (c) 2006 Elsevier B.V. All rights reserved.

The use of glucan as immunostimulant in the treatment of paracoccidioidomycosis

A group of 10 patients, nine of them seriously infected with Paracoccidioides brasiliensis (G1), received glucan (beta-1,3 polyglucose) as an immunostimulant intravenously once a week for one month, followed by monthly doses (10 mg) over an ii-month period, together with a specific anti-fungal agent as an immunostimulant. A second group of eight moderately infected patients (G2) was treated with only the anti-fungal agent. Among the patients in G1, there was only one case of relapse compared with five in G2. Values for the erythrocyte sedimentation rate (ESR) showed a significant difference (P < 0.01) post-treatment in G1 patients, when compared with the pretreatment levels. There was also a significant reduction (P < 0.001) in the level of serum antibodies to P. brasiliensis in the G1 patients in post-treatment examinations. The phytohemagglutinin (PHA) skin test showed a positive reaction among the patients in G1 (P < 0.01) post-treatment and there was a tendency towards an increase in the number of CD4+ T lymphocytes in both groups after treatment. The serum level of tumor necrosis factor (TNF) proved to be significantly higher (P < 0.02) in the G1 patients during treatment. In the G1 patients, the correlation between ESR and TNF tended to be negative whereas that between ESR and serum antibodies was positive. The present results indicate that the patients who received glucan, in spite of being more seriously ill, had a stronger and more favorable response to therapy.

Effect of grazing system on fetal development in Nellore cattle

Intensive grazing systems for beef females, based on abundant availability of high quality forages and supplementary concentrates, may affect fetal development. The objective of this study was to determine the effect of grazing system on length of gestation, fetal development, and characteristics of the calf at birth. Twenty-four pregnant (bred to Nellore bulls) Nellore females were allocated into two groups. The control group (G1) grazed Brachiaria decumbens (signal grass) in a traditional (extensive) grazing system and the second group (G2) were managed on Panicum maximumcv. Tanzania 1 (Tanzania grass) in an intensive grazing system. Fetal development was evaluated by ultrasonography on days 31, 45, 59, 94, 122, 220, and 255 of gestation. The diameter of the amniotic and allantoic cavities, crown-rump length, circumference, and diameter of the head and ocular orbit were determined. At birth, calves were weighed and height, length, thoracic circumference, and ocular orbit and bi-parietal diameters were measured. There were no differences (P > 0.05) in fetal development. The G1 cows had a longer gestation period (4.5 days; P < 0.05) and their calves had greater (P < 0.05) weight, height, length, and thoracic circumference at birth. In conclusion, Nellore females raised under intensive pasture management conditions, had significantly shorter gestation and smaller calves at birth than those raised under extensive pasture management conditions. Therefore, adoption of new management practices (e.g. intensive pasture management), should take into consideration animal behavior and productivity. (C) 2003 Elsevier B.V. All rights reserved.

INFLUENCE OF DIABETES-MELLITUS ON THE GLUTATHIONE REDOX SYSTEM OF HUMAN RED-BLOOD-CELLS

Several components of the erythrocyte-dependent glutathione redox system (reduced glutathione, GSH; oxidized glutathione, GSSG; glutathione peroxidase, GSH-Px; glutathione reductase, GSH-Red) were determined in patients with types I and II diabetes mellitus (DM). All groups studied were male subjects: G1, 20 young healthy individuals (aged 23.7 +/- 4.2 years); G2, 15 young insulin-treated type I DM patients; G3, 20 older insulin-treated type II DM patients; 04, 21 older oral hypoglycemic agent-treated type II DM patients; G5, 28 aged healthy individuals (aged 68.9 +/- 11.5 years). There were no differences between G1 and G2, G3 or G4 regarding erythrocyte GSH, GSSG, and GSH-Red (without FAD) levels. GSH-Px activity was significantly lower in G2 when compared to G1 (15.2 +/- 4.9 vs 20.6 +/- 6.6 IU/g Hb). The GSH-Red and GSH-Px activities and GSH levels were significantly higher in 03 (4.6 +/- 1.7 IU/g Hb, 20.2 +/- 8.7 IU/g Hb and 3.5 +/- 1.3-mu-M/g Hb) and G4 (5.0 +/- 2.2 IU/g Hb, 16.9 +/- 6.1 IU/g Hb and 5.0 +/- 2.3-mu-M/g Hb) when compared to G5 (3.4 +/- 0.9 IU/g Hb, 12.0 +/- 3.6 IU/g Hb and 2.3 +/- 0.9-mu-M/g Hb). The findings suggest that treatment of DM can stimulate the redox activity of red blood cells in aged subjects.

Effect of different glycosaminoglycans in a guinea-pig carotid-artery thrombosis model

Heparin is the most frequently used drug for the prevention and treatment of thrombosis. Its use, however, is restricted by its side-effects. To study the efficacy of other glycosaminoglycans that could substitute heparin in the management of arterial thrombosis, 60 guinea-pigs were randomly allocated into 6 groups: G1= control, G2= heparin (150 IU/kg), G3= heparan sulfate from beef pancreas (2.5 mg/kg), G4= heparan sulfate from beef lung (2.5 mg/kg), G5= N-acetylated heparan from beef pancreas, G6= dermatan sulfate from beef intestine (2.5 mg/kg). Ten minutes after intravenous injection of the drugs, thrombosis was induced by the injection of a 50% glucose solution into a segment of the right carotid artery isolated between 2 thread loops during 10 minutes. Three hours later the artery was re-exposed and if a thrombus was present it was measured, withdrawn and weighed. Thrombin time and activated partial thromboplastin time were measured in all animals. Thrombus developed in 90% of the animals in the control group, 0% in G2 and G3, 62.5% in G4, 87.5% in G5 and G6. Only in the animals treated with heparin the coagulation tests were prolonged. In conclusion, in the used dose only the heparan sulfate from beef pancreas presented an antithrombotic effect similar to heparin in this experimental model.

Neurotoxicity of subarachnoid hyperbaric bupivacaine in dogs

Background and Objectives. The study investigated possible neurotoxic effects of increasing concentrations and doses of bupivacaine administered into the subarachnoid space in dogs. Methods. Fifty animals were allocated to five experimental groups: G1, control; G2, 5 mg 0.5 bupivacaine in 10% glucose solution; G3, 10 mg of 1% bupivacaine in 10% glucose solution; G4, 20 mg 2% bupivacaine in 10% glucose solution, and G5, 20 mg 2% bupivacaine in water. After 72 hours of observation, the animals were killed and the spinal cords removed for histologic examination by light microscopy. Results. None of the animals showed any neurologic clinical disturbance following recovery from spinal anesthesia. One case of necrosis of nerve tissue was observed in G3 and four in G4. Conclusions. Increasing concentrations and doses of hyperbaric bupivacaine solutions increased the incidence of nerve tissue damage, which did not occur with hypobaric solutions. These results should contribute to the further understanding of neurologic complications following spinal anesthesia when large doses of local anesthetics in hyperbaric solutions are used.

CYTOMETRY AND KARYOMETRY IN TRYPANOSOMA-CRUZI

A total of 991 Trypanosoma cruzi cells, from four laboratory stocks, including the three differentiation forms, had their cellular outlines, nuclei and kinetoplasts measured at 9000 x magnification. Data on the identifiable cell cycle stages were used to search for intraspecific and biological cycle heterogeneity.Cellular areas (CA) in the interphasic differentiation forms produced ratios of 1.07 for culture epimastigotes (E), 1 for blood trypomastigotes (T), and 0.86 for tissue forms (A). Homogeneity in terms of nuclear (NA) and kinetoplast (KA) areas prevailed among the stocks, with differences of at most 6%, for modal NA of strains CL and Y. NA of T-form was larger than the basic NA of early G1 A-form. T-form kinetoplast volume was 3-fold that of A-form K-DNA nucleoids.One of the two recently divided kinetoplasts in mitotic E-form did not correlate with CA, indicating that mitochondrial division was unequal. The KA of CL strain T-form did not correlate with NA, suggesting a mitochondrial disfunction in this thermosensitive strain.The CL strain T-form was more heterogeneous than the Y strain for all characters, showing greater frequency of large values, even reaching the G2 levels. This heterogeneity was interpreted as functional, consequent to the thermosensitivity of the CL strain. Precocious bursting of CL strain host cells would lead to the polymorphic T-forms. Post-S phase trypomastigotes could start division soon after penetration of host cells.

Enamel caries resistance accidentally irradiated by the Nd : YAG laser

When the carious tissue is eliminated either by conventional methods (with burs) or with lasers, the risk of accidentally damage the surface of adjacent teeth may occur, which hypothetically could lead to a more susceptible surface for canes formation. This in vitro study aims to evaluate the caries resistance of the dental enamel surface irradiated by the Nd:YAG laser applied in conditions simulating accidental exposition. Thirteen third molars were used in this study. The experimental groups were: G1: sound control and control + carious; G2: contact Nd:YAG laser at 0.75, 1, 2, or 3 W; 10 Hz; 3 sec (27, 35, 71, and 106 J/cm(2)); G3: same parameters from G2 + caries artificial induction through the demineralization and demineralization (DES/RE) dynamic model. The caries resistance analysis was evaluated by the superficial morphological aspect through SEM images and also by Ca/P proportion through energy dispersive X-ray spectroscopy (EDX). The micrograph images showed that the Nd:YAG laser changed the normalmorphology of the enamel prisms resulting in a melted and re-solidified surface intensified with the power increase. Significant statistical differences were observed applying the Kruskal-Wallis statistical test (p <= 0.01) among the Nd:YAG laser irradiated groups and the control with caries regarding the Ca/P proportion. As an exception, this was not observed when 3 W; 10 Hz; 3 sec; 106 J/cm(2) was applied and posteriously submitted to a cariogenic challenge. The results indicate that the Nd:YAG laser accidental irradiation at low power settings did not represent risks to the enamel caries resistance.

Lidocaine effects on corneal endothelial cell ultrastructure

The purpose of this study was to determine whether intracameral commercial lidocaine 2% induces alterations on the rabbit corneal endothelium. Forty white rabbits received different substances inside the anterior chamber: group (G)1, no substance; G2 and G3 received lidocaine 2% with preservative in aqueous solution; G4 and G5, lidocaine 2% with preservative in gel solution; G6 and G7, the anesthetic preservative (metilparahydroxybenzoate 0.1%); and G8 and G9, lidocaine 2% without preservative in aqueous solution. The animals from G2, 4, 6 and 8 were sacrificed after 1 h, and from G3, 5, 7 and 9 after 24 h after injection of the substance inside the anterior chamber. The corneas were clinically evaluated and assessed by transmission and scanning electron microscopy. G1, 2, 6, 7, 8 and 9 animals had very similar characteristics in clinical, ultrastructural and morphometric evaluations; the G3 and G4 animals showed discrete edema and one animal in G5 had intense corneal edema. We conclude that lidocaine 2% with preservative induces few ultrastructural alterations in the corneal endothelial cells.

Temperature rise in cavities prepared by high and low torque handpieces and Er : YAG laser

Objective The aim of this study was to compare intrapulpal temperature increases produced by a high-speed high-torque (speed-increasing) handpiece, a high-speed low-torque handpiece (air-turbine) and an Er: YAG (Erbium: Yttrium-Aluminum-Garnet) laser. Subject and methods Thirty bovine incisors were reduced to a dentine thickness of 2.0 mm. Class V preparations were prepared to a depth of 1.5 mm, measured with a caliper or by a mark on the burs. A thermocouple was placed inside the pulp chamber to determine temperature increases (C). Analysis was performed on the following groups (n = 10) treated with: G1, low-torque handpiece; G2, high-torque handpiece; and G3, Er: YAG laser (2.94 mu m at 250 mJ/4 Hz), all with water cooling. The temperature increases were recorded with a computer linked to the thermocouples. Results The data were submitted to ANOVA and Tukey statistical test. The average temperature rises were: 1.92 +/- 0.80 degrees C for G1, 1.34 +/- 0.86 degrees C for G2, and 0.75 +/- 0.39 degrees C for G3. There were significant statistical differences among the groups (p = 0.095). All the groups tested did not have a change of temperature that exceeds the threshold of 5.5 degrees C. Conclusion Temperature response to the low and high torque handpieces seemed to be similar, however the Er: YAG laser generated a lower temperature rise.

LIFE QUALITY of PEOPLE INFECTED BY HIV WITH OR WITHOUT ANTIRETROVIRAL TREATMENT

The "HIV/Aids-Quality of Life" (HAT-Qol) is a specific multifunctional instrument used to measure the life quality of HIV infected persons. It is divided into nine domains: general activity, sexual activity, secrecy about HIV seropositivity, concern about health, financial concern, awareness about HIV, satisfaction with life, issues about medications and belief in the doctor. The current study analyzed the life quality of HIV infected individuals-who attended the DST/Aids Program in Maringa city, Parana state-regarding the use or not of antiretroviral therapy (TARV) and their demographic, epidemiological and clinical characteristics. Data were collected by retrospective analysis from 1,200 medical charts of patients registered in the program. The HAT-Qol instrument was applied before routine medical consultation. One hundred and sixty-nine patients, who had HIV infection confirmed, were divided into two groups, G1 with 118 individuals receiving antiretroviral therapy and G2 with 51 individuals who were not under this therapy.Result analysis, regarding social and demographic characteristics, revealed no difference among responses related to gender, educational degree and sexual option. Age influenced satisfaction with sexual activity and marital status. Regarding HIV awareness, the lowest response index or worst quality of life came from, respectively, men between 50 and 69 years old and patients who did not have regular partners compared with the ones who did. Additionally, it was observed that the time of diagnosis influenced general activities, HIV awareness, concern about health and financial issues, satisfaction with life and topics about medications. The variables were compared in both groups. There was no influence on the use or not of antiretroviral therapy regarding age, sexual activity, HIV diagnosis time and the domains that evaluated general activities, financial concern, awareness of HIV and satisfaction with life. In relation to time of diagnosis, there was an influence only in persons who had been diagnosed two or three years before, in which a lower quality of life was observed among individuals who were not under antiretroviral therapy. It was not possible to compare variables about medication use, HIV plasmatic viral rate ant time of diagnosis, because G2 individuals were not receiving the antiretroviral therapy. Furthermore, no comparison was made regarding marital status and HIV awareness, because there were no married individuals in G2. Thus, the analysis of the results showed that the use of antiretroviral treatment did not influence the life quality of HIV patients studied by the HAT-Qol scale.

Población folicular y concentraciones plasmáticas de progesterona (P4) en ovejas sometidas a diferentes protocolos de sincronización

Los efectos de la prostaglandina (PGF2α) vs CIDR y eCG (gonadotrofina coriónica equina) en la dinámica de la población folicular y su relación con las concentraciones plasmáticas de P4 fueron investigadas en ovejas cíclicas. Fueron utilizadas 14 hembras ovinas de la raza Bergamascia; el Grupo 1 (Gl) se sometió a dos aplicaciones de PGF2α, y, el Grupo 2 (G2) tratado con CIDR durante 14 días siendo que, en el momento de su retirada, se administraron 500 UI de eCG. La dinámica folicular ovárica fue monitoreada por medio de ecógrafo. Se monitorearon todos los folículos ≥ mm y se gráfico su posición diariamente, observándose el desarrollo individual folicular. Desde el día anterior a la aplicación de la segunda dosis de PGF2α, (Gl) y desde la administración de eCG (G2) hasta el décimo día del ciclo estral se colectaron muestras de sangre para el análisis de P4. Hubo diferencia significativa (P<0,001) en las concentraciones plasmáticas de P4 entre los tratamientos. La sincronización del estro y ovulación utilizando CIDR + 500 UI de eCG, incrementó la cantidad de folículos reclutados, además de aumentar el diámetro máximo y la tasa de crecimiento de los folículos grandes en la primera onda folicular. En consideración a los resultados se puede concluir que la sincronización del estro y de la ovulación en hembras ovinas, utilizando el CIDR y 500 UI de eCG, incrementa la cantidad de folículos reclutados, además de aumentar el diámetro máximo y la tasa de crecimiento de los folículos grandes. La asociación CIDR+500 UI de eCG provoca aumentos significativos en las concentraciones plasmáticas de progesterona (P4) al inicio de la fase luteal en hembras ovinas.

Infestation of Bemisia tabaci (Genn.) biotype B (Hemiptera: Aleyrodidae) and Caliothrips phaseoli (Hood.) (Thysanoptera: Thripidae) in beans genotypes.

The objective of this work was to evaluated the infestion of the whitefly Bemisia tabaci (Genn.) B biotype and thrips Caliothrips phaseoli (Hood) in fifteen beans genotypes in field condition. The genotypes IAPAR 31, Rosinha G2, Jalo precoce, Perola, IAC Harmonia, Gen 99TGR110, Gen 99TG2868, Gen 99TGR3416, Gen 99TG3450, Gen 99TG823, Gen 99TGR609, IAC Jaragua, Gen 95A10061531, Gen 99TGR3114 e Gen 96A1473153V2 was used. The experiment was conducted from May to July 2006. The statistical design was randomized blocks, totalizing 15 treatments and four replications. Evaluation was realized weekly, totalizing seven samplings. The evaluations were accomplished on a weekly basis by counting B. tabaci biotype B eggs and nymphs and nymphs of C. phaseoli in 10 leaflets per plot. The less oviposition genotypes by B. tabaci biotype B were IAC Harmonia, Perola, Gen TG3114 e Gen 95A10061531, while the most oviposited were IAC Jaragua and Gen 99TG3450. The less presence of nymphs of whitefly were observed on Perola and IAC Harmonia and the most at IAC Jaragua. It was observed a negative and non significant linear correlation between average temperature versus number of silverleaf whitefly and temperature versus number of thrips was verified.

The induction of cytotoxicity, oxidative stress, and genotoxicity by root canal sealers in mammalian cells

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