Immunohistochemical Detection of Intralesional Antigens of Ovine Gammaherpesvirus-2 in Cattle with Sheep-associated Malignant Catarrhal Fever.

Sheep-associated malignant catarrhal fever (SA-MCF) is a severe lymphoproliferative disease of ruminants caused by ovine gammaherpesvirus-2 (OvHV-2). Since the initial identification of SA-MCF there has been extensive research related to the pathogenesis of OvHV-2, based primarily on serological and molecular assays associated with typical histopathological findings. The monoclonal antibody (MAb-15A) binds to a common epitope in MCF viruses and is used frequently in serological investigations. However, the utilization of this antibody to detect antigens of OvHV-2 in tissues has not been examined. Accordingly, this study standardized an immunohistochemical assay using MAb-15A to identify antigens of OvHV-2 in tissues of cattle (n = 5) with SA-MCF. All animals developed acute neurological signs, without ocular and nasal manifestations, and had nucleic acids of OvHV-2 in brain tissue detected by polymerase chain reaction. The principal histopathological findings were lymphocytic nephritis (n = 5), widespread arterial proliferation and vasculitis (n = 5), lymphocytic portal hepatitis (n = 3), non-suppurative meningoencephalitis (n = 2) and atrophic enteritis with cryptal necrosis and dilation (n = 2). Intralesional intracytoplasmic antigens of OvHV-2 were identified within multiple epithelial cells of the kidneys of all animals, the intestines of animals with and without atrophic enteritis, and within epithelial cells of bile ducts in animals with lymphocytic hepatitis. Additionally, there was positive intracytoplasmic immunoreactivity within histiocytes and lymphocytes in several tissues. These findings suggest that the MAb-15A detects antigens of OvHV-2 within epithelial cells and leucocytes in several organs. Moreover, this assay would contribute significantly towards understanding of the pathogenesis of SA-MCF and may be used for retrospective studies. Additionally, angiopathy in SA-MCF may be a progressive lesion, which may terminate in luminal occlusion and probably occurs irrespectively of the eye and head form of MCF.

Prevalence of foot disorders in lactating Jersey cows raised in semi-confinement.

BACKGROUND AND AIM: To establish prevention strategies, recording the prevalence of foot injuries within a herd should be the starting point in determining the risk factors involved in digital diseases. This study aimed to assess the prevalence of claw disorders in lactating Jersey cows raised in a semi-confinement system. MATERIALS AND METHODS: Five hundred and eighty-four digits were examined from 73 lactating Jersey cows. The lameness score system was used to assess each cow while walking and on standing position, and digital lesions were evaluated with the cows restrained in a hydraulic cattle chute. RESULTS: The prevalence of digital lesions was 93.1%. Among the 68 affected cows, only 21 were lame. Of the 584 digits examined, 970 lesions were recorded, corresponding to 13.3% lesions per cow and 1.66% injuries per digit. Forty-eight cows (65.7%) had lesions in all digits, and 92.8% of digits had at least one lesion. Lesions in digits of fore limbs were more common (55.3%) (p<0.0001) than those of hind limbs (44.7%). Foot injuries in medial digits of the fore limbs were more prevalent (56.8%) (p<0.001) than in the lateral digits (43.2%). The lesions' occurrence was similar in both medial and lateral digits of the hind limbs (p=0.8347). The primary diseases observed were heel horn erosion (53.8%), white line disease (19.3%), and double sole (12.4%), which together accounted for 92.4% and 84.9% of foot disorders diagnosed in the fore and hind limbs, respectively. Other digital diseases occurred less frequently. CONCLUSION: The prevalence of foot disorders in lactating Jersey cows raised in semi-confinement systems was high. This could be due to the lack of preventive trimming, infectious diseases, and nutritional problems.

Apparent selectivity of sheep in deferred marandu palisadegrass pastures with variable initial heights / Seletividade aparente de ovinos em pastos de capim-marandu diferidos com alturas iniciais variáveis

The objective of this study was to evaluate the apparent selectivity of sheep in marandu palisadegrass (Urochloa brizantha cv. Marandu) pastures with four heights at the beginning of the deferment period (15, 25, 35 and 45cm). The deferment period was 92 days and started on 03/21/2014. Evaluations occurred in the beginning (first week), middle (45th day) and end (92nd day) of the grazing period, in winter (06/21/2014 to 09/21/2014). Deferred pastures with 15 and 25cm presented lower forage mass (FM), but higher live leaf (LL) percentage in FM than deferred pastures with 35 and 45cm. The live stem percentage in the FM and the apparent selectivity index (ASI) of the LL were superior in the deferred pasture with 45cm. The dead stem (DS) percentage in the grazing simulation (GS) and the ASI of this morphological component were lower in the pasture with 15cm, compared to the deferred pasture with 45cm. The FM and the LL percentages in FM and in the GS sample decreased, while the DS percentages in FM and in GS sample increased with the grazing period. Marandu palisadegrass with 15cm at beginning of the deferment period improves the morphology of the deferred pasture. Selective grazing is difficult during the grazing period.(AU)

Objetivou-se avaliar a seletividade aparente de ovinos em pastos de capim-marandu (Urochloa brizantha cv. Marandu) com quatro alturas no início do diferimento (15, 25, 35 e 45cm). O período de diferimento foi de 92 dias e iniciou em 21/03/2014. As avaliações ocorreram no início (primeira semana), meio (45° dia) e fim (92° dia) do período de pastejo, no inverno (21/06/2014 a 21/09/2014). Os pastos diferidos com 15 e 25cm apresentaram menor massa de forragem (MF), mas maior percentual de folha viva (FV) na MF do que os pastos diferidos com 35 e 45cm. O percentual de colmo vivo na MF e o índice de seletividade aparente (ISA) da FV foram superiores no pasto diferido com 45cm. O percentual de colmo morto (CM) na simulação de pastejo (SP) e o ISA desse componente morfológico foram menores no pasto diferido com 15cm, em comparação ao diferido com 45cm. A MF e os percentuais de FV na MF e na amostra de SP se reduziram, enquanto os percentuais de CM na MF e na amostra de SP aumentaram com o período de pastejo. O capim-marandu com 15cm no início do período de diferimento melhora a morfologia do pasto diferido. O pastejo seletivo é dificultado no decorrer do período de pastejo.(AU)

PCR-based identification of Mycobacterium murphy causing Canine Leproid Granuloma Syndrome in Niterói, southeast Brazil - case report / Identificação baseada em PCR de Mycobacterium murphy causando Síndrome do Granuloma Lepróide Canino em Niterói, sudeste do Brasil ˗ Relato de Caso

Canine Leproid Granuloma Syndrome (CLGS), also known as canine leprosy, is a cutaneous nodular infectious disease caused by Mycobacterium sp.. Despite being reported worldwide, it is still quite unknown and underdiagnosed. Diagnosis may be achieved by cytopathology or histopathology of skin lesions, but identification of the infectious agent is complex, since bacterial in vitro growth is not possible, relying upon molecular techniques such as PCR to confirm Mycobacterium DNA in the sample. We report a CLGS case in Niteroi, Rio de Janeiro state, Brazil, diagnosed by cytopathology and submitted to molecular identification of the agent. PCR amplification of hsp65 gene was performed and revealed 100% genetic homology to M. murphy strain. This is the first CLGS report with molecular identification in Rio de Janeiro state, and this finding should raise awareness about CLGS as a differential diagnosis among granulomatous skin diseases in this region.(AU)

A síndrome de granuloma leproide canino (SGLC), também conhecida como lepra canina, é uma doença infecciosa cutânea nodular causada por Mycobacterium sp. Apesar de ser relatada mundialmente, ainda é bastante desconhecida e subdiagnosticada. O diagnóstico pode ser conseguido por citopatologia ou histopatologia de lesões cutâneas, mas a identificação do agente infeccioso é complexa, uma vez que o crescimento in vitro bacteriano não é possível, dependendo de técnicas moleculares como a PCR para confirmar o DNA de Mycobacterium na amostra. Relatou-se um caso da SGLC em Niterói, estado do Rio de Janeiro, Brasil, diagnosticado por citopatologia e submetido à identificação molecular do agente. Foi realizada amplificação por PCR do gene hsp65, que revelou 100% de homologia genética com a cepa M. murphy. Este é o primeiro relato da SGLC com identificação molecular no estado do Rio de Janeiro, o que mostra a importância de se acrescentar a SGLC ao diagnóstico diferencial das doenças granulomatosas de pele nessa região.(AU)

IL33 and IL1RL1 variants are associated with asthma and atopy in a Brazilian population.

Atopic asthma is a chronic inflammatory disease in airways resulting from genetic and environmental factors, characterized by production of the Th2 cytokines interleukin-4 (IL-4), interleukin-5 (IL-5) and interleukin-13 (IL-13). Interleukin-33 (IL-33) appears to be a potent inducer of Th2 immune response. This occurs when IL-33 binds and activates its receptor, the membrane ST2 (ST2L) in mast cells, dendritic cells, basophils, eosinophils, innate lymphoids and Th2 cells, leading to the release of these cytokines and intensifying allergic inflammation. Polymorphisms in the IL33 and IL1RL1 can act as protective or risk factors for asthma and/or allergy in humans. No study was conducted to replicate such findings in a European and African descendent mixed population. DNA was extracted from peripheral blood from 1223 subjects, and the samples were genotyped using Illumina 2.5 Human Omni Beadchip. We tested for possible associations between SNPs in the IL33 and ST2 with asthma and allergy markers such as specific IgE (sIgE), IL-5 and IL-13 production and skin prick test (SPT). Logistics regressions were performed using PLINK software 1.07. The analyses were adjusted for sex, age, helminth infection and ancestry markers. The G allele of IL33 SNP rs12551256 was negatively associated with asthma (OR 0.71, 95% CI: 0.53-0.94, P = 0.017). In contrast, the A allele of IL1RL1 rs1041973 was positively associated with IL-5 production (OR 1.36, 95% CI: 1.09-1.84, P = 0.044), sIgE levels (OR 1.40, 95% CI: 1.07-1.84, P = 0.013) and positive SPT (OR 1.48, 95% CI: 1.08-2.03, P = 0.014), for Blomia tropicalis mite. The same allele, in atopic subjects, was associated with decreased production of soluble ST2 (sST2) (P < 0.05). Moreover, expression quantitative trait loci (eQTL) analysis suggests that rs1041973 and rs873022 regulate the expression of IL1RL1 gene. This latest SNP, rs873022, the T allele, was also associated with a lower production of sST2 in plasma of Brazilians. The genetic risk score for rs1041973 and rs16924161 demonstrated a higher risk for SPT positivity against B. tropicalis, the greater the number of risk alleles for both SNPs. Our findings demonstrate a robust association of genetic variants in IL1RL1 and IL33 SNPs with allergy markers and asthma.

Characterization of the gene encoding component C3 of the complement system from the spider Loxosceles laeta venom glands: Phylogenetic implications.

A transcriptome analysis of the venom glands of the spider Loxosceles laeta, performed by our group, in a previous study (Fernandes-Pedrosa et al., 2008), revealed a transcript with a sequence similar to the human complement component C3. Here we present the analysis of this transcript. cDNA fragments encoding the C3 homologue (Lox-C3) were amplified from total RNA isolated from the venom glands of L. laeta by RACE-PCR. Lox-C3 is a 5178 bps cDNA sequence encoding a 190kDa protein, with a domain configuration similar to human C3. Multiple alignments of C3-like proteins revealed two processing sites, suggesting that Lox-C3 is composed of three chains. Furthermore, the amino acids consensus sequences for the thioester was found, in addition to putative sequences responsible for FB binding. The phylogenetic analysis showed that Lox-C3 belongs to the same group as two C3 isoforms from the spider Hasarius adansoni (Family Salcitidae), showing 53% homology with these. This is the first characterization of a Loxosceles cDNA sequence encoding a human C3 homologue, and this finding, together with our previous finding of the expression of a FB-like molecule, suggests that this spider species also has a complement system. This work will help to improve our understanding of the innate immune system in these spiders and the ancestral structure of C3.

Presença do genoma de BoHV-5 no líquido cefalorraquidiano de bovinos com meningoencefalite herpética / Presence of BoHV-5 genome in cerebrospinal fluid of cattle with herpetic meningoencephalitis

The diagnosis of bovine herpesvirus 5 (BoHV-5) encephalitis is confirmed after death by laboratory methods applied to brain fragments. Alternative methods to confirm ante-mortem diagnosis are important because the disease is not always lethal. The aim of this study was to investigate whether the presence of the virus genome in the cerebrospinal fluid (CSF) detected by polymerase chain reaction (PCR) might be admitted as a method for ante-mortem diagnosis. CSF samples were taken from 14 animals suffering from BoHV-5 encephalitis, diagnosed by characteristic histopathological lesions in the brain and by identification of the virus genome by PCR in different portions of the brain. Virus DNA was detected in the CSF of 21.42% (3/14) of the evaluated animals. Ante-mortem detection of the virus genome in the CSF showed low sensitivity to confirm the diagnosis. The diagnosis is confirmed by a positive result but a negative one does not discard the disease.(AU)

The interrelation between aPKC and glucose uptake in the skeletal muscle during contraction and insulin stimulation.

Contraction and insulin increase glucose uptake in skeletal muscle. While the insulin pathway, better characterized, requires activation of phosphoinositide 3-kinase (PI3K) and atypical protein kinase (aPKC), muscle contraction seems to share insulin-activated components to increase glucose uptake. This study aimed to investigate the interrelation between the pathway involved in glucose uptake evoked by insulin and muscle contraction. Isolated muscle of rats was treated with solvent (control), insulin, wortmannin (PI3K inhibitor) and the combination of insulin plus wortmannin. After treatment, muscles were electrically stimulated (contracted) or remained at rest. Glucose transporter 4 (GLUT4) localization, glucose uptake and phospho-aPKC (aPKC activated form) were assessed. Muscle contraction and insulin increased glucose uptake in all conditions when compared with controls not stimulating an effect that was accompanied by an increase in GLUT4 and of phospho-aPKC at the muscle membrane. Contracted muscles treated with insulin did not show additive effects on glucose uptake or aPKC activity compared with the response when these stimuli were applied alone. Inhibition of PI3K blocked insulin effect on glucose uptake and aPKC but not in the contractile response. Thus, muscle contraction seems to stimulate aPKC and glucose uptake independently of PI3K. Therefore, aPKC may be a convergence point and a rate limit step in the pathway by which, insulin and contraction, increase glucose uptake in skeletal muscle.

Characterization of anti-crotalic antibodies.

Crotalus durissus terrificus, C. d. collilineatus, C. d. cascavella and C. d. marajoensis are responsible minor but severe snake bites in Brazil. The venoms of these snakes share the presence of crotoxin, a neurotoxin comprising of two associated components, crotapotin and phospholipase A2 (PLA2). Treatment of the victims with specific antiserum is the unique effective therapeutic measure. The ability of anti-Crotalus antisera produced by the routine using crude venom to immunize horses or purified crotoxin and PLA2 as individual immunogens was compared. Antisera obtained from horses immunized with C. durissus terrificus crude venom were able to recognize and neutralize not only the toxins presents in C. durissus terrificus, but also the ones present in the venoms from C. d. collilineatus, C. d. cascavella and C. d. marajoensis. Antisera from horses immunized with individual crotoxin or PLA2, although in lesser titers, were also able of recognizing the toxins in all four Crotalus species and neutralize the lethality of the C. d. terrificus venom.

Serum vascular endothelial growth factor in dogs with soft tissue sarcomas.

This work aimed to evaluate serum vascular endothelial growth factor (VEGF) in 25 dogs with soft tissue sarcoma, and in 30 healthy dogs. Blood was collected once time from the control animals and three times, in the same way, from animals with sarcoma. Blood count was performed in the blood collected, and serum VEGF was measured by enzyme-linked immunosorbent assay quantitative method. Serum VEGF in control animals was similar to patients with soft tissue sarcoma. There was a reduction in serum VEGF after the sarcoma resection. There was positive correlation between serum VEGF and neutrophil counts, and negative between VEGF and hemoglobin content in animals with sarcoma. Animals with hemangiopericytoma showed higher serum VEGF levels compared to the patients with malignant peripheral nerve sheath. Circulating blood cells can contribute to elevate VEGF serum concentrations in dogs with soft tissue sarcomas and a possible role of VEGF in the angiogenesis of these tumors.

Effect of pentoxifylline on the regeneration of rat testicular germ cells after heat shock

High temperature is an important factor for reproduction and can induce testicular degeneration. Pentoxifylline is a methylxanthine phosphodiesterase inhibitor with anti - inflammatory and anti - apoptotic properties. Considering the protective propert ies of pentoxifylline and the harmful effects of heat, the present study aimed to use pentoxifylline to reduce the damage induced by heat to the testis. Adult mal e Wistar rats were exposed to testicular heat shock (43ºC for 15 min) , treated with 50 or 100 mg/ k g of pentoxifylline and evaluated at 3, 7, 15, 30 and 60 days after heat shock. Pentoxifylline treatment did not change testicular weight, histomorphometrical parameters or plasma testosterone concentration. However, pentoxifylline inhibited germ cell apoptosis and reduced the severity of pathological lesions at 30 and 60 days after testicular heat shock. In conclusion, pentoxifylline treatment seem ed to inhibit pro - inflammatory and apoptotic mechanisms triggered by testicular heat shock, improving sper matogenesis regeneration.

Eating behavior and nutritional status in patients who underwent coronary angioplasty.

INTRODUCTION: The identification of stages of dietary change and the factors affecting food choices can direct more effective nutritional intervention against coronary heart disease progression. OBJECTIVE: Identify the stages of change of eating behavior and its relation with nutritional status, food consumption and previous cardiovascular events in patients who underwent coronary angioplasty. METHODS: A cross-sectional study with 200 hospitalized patients from a specialized cardiology hospital, after elective coronary angioplasty. They were applied an algorithm that identifies the provision of change of eating habits for a healthier pattern. Variables measured were stages of change of eating behavior, nutritional status, food consumption and cardiovascular events (previous myocardial infarction or angioplasty). It was realized comparison of averages by analysis of variance or Student's test and Chi-square test for qualitative variables. Value of significance was taken at 5%. RESULTS: The patients were classified in the following stages: 36% maintenance, 26% preparation, 17% precontemplation, 12% action and 9% contemplation. It was observed higher cardiovascular events in maintenance/action group (p = 0.04), higher consumption of calories (p = 0.04), meat/eggs (p = 0.01) and sweets (p = 0.03) in preparation stage, comparing to maintenance group, and no association between nutritional status and stages of change (p = 0.13), although 62% of the individuals in maintenance stage were overweight. CONCLUSIONS: This work contributed to identifying the stages of change and conditions that favor changes in eating pattern. Even patients that classified themselves into the maintenance stage need to adjust their eating habits in order to reach a healthy weight.

Cerebral cryptococcomas in a cow.

Cerebral cryptococcomas are described in a 5-year-old mixed-breed cow without manifestations of systemic cryptococcosis. Two cryptococcomas were observed grossly. Microscopical examination revealed accumulations of yeast that were morphologically consistent with Cryptococcus neoformans. Immunohistochemistry characterized the organisms as C. neoformans var. grubii.

Eating behavior and nutritional status in patients who underwent coronary angioplasty / La conducta alimentaria y estado nutricional en pacientes sometidos a angioplastia coronaria

Introduction: The identification of stages of dietary change and the factors affecting food choices can direct more effective nutritional intervention against coronary heart disease progression. Objective: Identify the stages of change of eating behavior and its relation with nutritional status, food consumption and previous cardiovascular events in patients who underwent coronary angioplasty. Methods: A cross-sectional study with 200 hospitalized patients from a specialized cardiology hospital, after elective coronary angioplasty. They were applied an algorithm that identifies the provision of change of eating habits for a healthier pattern. Variables measured were stages of change of eating behavior, nutritional status, food consumption and cardiovascular events (previousmyocardial infarction or angioplasty). It was realized comparison of averages by analysis of variance or Student’s test and Chi-square test for qualitative variables. Value of significance was taken at 5%.Results: The patients were classified in the following stages: 36% maintenance, 26% preparation, 17% precontemplation, 12% action and 9% contemplation. It was observed higher cardiovascular events in maintenance/action group (p = 0.04), higher consumption of calories (p =0.04), meat/eggs (p = 0.01) and sweets (p = 0.03) in preparation stage, comparing to maintenance group, and no association between nutritional status and stages of change (p = 0.13), although 62% of the individuals in maintenance stage were overweight. Conclusions: This work contributed to identifying the stages of change and conditions that favor changes in eating pattern. Even patients that classified themselves in to the maintenance stage need to adjust their eating habits in order to reach a healthy weight (AU)

Introducción: La identificación de las etapas de cambio en la dieta y los factores que afectan la elección de alimentos puede dirigir una intervención nutricional más eficaz contra la progresión de la enfermedad coronaria. Objetivos: Identificar las etapas de cambio de la conducta alimentaria y su relación con el estado nutricional, consumo de alimentos y los eventos cardiovasculares anteriores en pacientes sometidos a angioplastia coronaria. Métodos: Estudio transversal con 200 pacientes de un hospital especializado en cardiología, después de angioplastia coronaria electiva. Se aplicaron un algoritmo que identifica la disposición del cambio de hábitos de alimentación para un patrón mas saludable. Las variables medidas fueron las etapas de cambio de la conducta alimentaria, estado nutricional, consumo de alimentos y eventos cardiovasculares (infarto de miocardio o angioplastia previos).Resultados: los pacientes fueron clasificados en las siguientes etapas: 36% en mantenimiento, 26% en preparación, 17% en precontemplación, 12% en acción y 9%en contemplación. Se observaron mas eventos cardiovasculares en mantenimiento/ acción (p = 0,04), mayor consumo de calorías (p = 0,04), carne y huevos (p = 0,01) y dulces (p = 0,03) en la etapa de preparación, en comparación con el de mantenimiento, y ninguna asociación entre el estado nutricional y las etapas de cambio (p = 0,13),aunque el 62% de los individuos en la etapa de mantenimiento estaban con sobrepeso. Conclusiones: Este trabajo contribuyó por la identificación de las etapas de cambio y las condiciones que favorecen los cambios en el patrón de alimentación. Mismo los pacientes que se clasificaron a la fase de mantenimiento hay la necesidad de ajustar sus hábitos alimenticios con el fin de lograr un peso saludable (AU)

The effect of age on glucose uptake and GLUT1 and GLUT4 expression in rat skeletal muscle.

During the life span, phenotypic and structural modifications on skeletal muscle contribute to a reduction on glucose uptake either in basal state or triggered by insulin, but the underlying mechanisms for this decline are not entirely identified. A reduction in the expression of skeletal muscle glucose transporters (GLUTs), glucose transporter type 1 (GLUT1) and glucose transporter type 4 (GLUT4), has been associated to such phenomena, but unlike the case of insulin, only few studies have addressed the effect of age on muscle-contraction-induced glucose uptake. The aim of the study was to investigate the influence of age on GLUT1 and GLUT4 expression in skeletal muscle and its relation to the glucose uptake induced by muscle contraction. For this purpose, soleus muscle from Wistar rats aged 4, 10, 22 and 42 weeks were isolated and electrically stimulated (30 min, 10 Hz, 20 V, 0.2 ms). After stimulation, glucose uptake and GLUT1 and GLUT4 expression and localisation were evaluated. Muscle contraction caused an increase in glucose uptake in all studied groups. In addition, the absolute rates of glucose uptake were negatively correlated with age. The expression of GLUT4 was lower in older animals, whereas no relation between age and GLUT1 expression was found. Immunohistochemistry confirmed the ontogenic effect on GLUT4 expression and suggested an age-related modification on GLUT1 distribution within the muscle fibres; for instance, this protein seems to be present mainly out of the sarcoplasm. The present findings demonstrate that the ability of muscle contraction to increase glucose uptake is not influenced by age, whereas glucose uptake under basal conditions decreases with age.

Opposite modulation of astroglial proliferation by adenosine 5'-O-(2-thio)-diphosphate and 2-methylthioadenosine-5'-diphosphate: mechanisms involved.

The contribution of P2Y(12,13) receptors to astroglial proliferation was investigated by testing the effects of two agonists with high affinity for these receptors, adenosine 5'-O-(2-thio)-diphosphate (ADPßS) and 2-methylthioadenosine-5'-diphosphate (2-MeSADP), in the incorporation of [(3)H]-thymidine. The effect of ATP, an endogenous inducer of astroglial proliferation, was also investigated. ADPßS and ATP (0.01-1 mM) increased astroglial proliferation up to 282%, an effect inhibited by the P2Y(1) receptor antagonist MRS 2179 (30 µM). The P2Y(12) receptor antagonists MRS 2395 (10 µM) and AR-C 66096 (10 µM) also reduced ADPßS proliferative effect, whereas the effect of ATP was attenuated by the A(2A) and A(2B) receptor antagonists SCH 58261 (30 nM) and MRS 1706 (10 nM), respectively. Studies of the signalling pathway activated showed that ADPßS effect was attenuated by pertussis toxin and by inhibition of phopholipase C (PLC), protein kinase C (PKC) and extracellular signal-regulated kinase1/2 (ERK1/2). The effect of ATP was also attenuated by inhibition of protein kinase A (PKA). The agonist 2-MeSADP (0.001-10 µM) had no effect in astroglial proliferation, but at higher concentrations (0.1-1 mM) it inhibited up to 63%, by mechanisms independent of P2Y(1,12,13) receptors activation. It was metabolised into 2-methylthioadenosine (2-MeSADO), the metabolite responsible for inhibition of astroglial proliferation. The effect of 2-MeSADO (0.1 mM) was attenuated by the A(3) receptors antagonist MRS 1523 (10 µM) and by the inhibitor of nucleoside transporters uridine (0.3 mM). 2-MeSADO did not induce apoptosis but increased lactate dehydrogenase release, an indicator of necrotic cell death. Astroglial proliferation induced by ADPßS was mediated by P2Y(1) and P2Y(12) receptors, leading to activation of PLC-PKC-ERK1/2 signalling pathway. The ATP proliferative effect was also mediated by PKA, supporting the contribution of the A(2) receptors. 2-MeSADP inhibition of astroglial proliferation depended on its conversion into 2-MeSADO, which activated A(3) receptors, blocked [(3)H]-thymidine uptake by astrocytes and led to cell death.

Prolina livre em alecrim-pimenta sob estresse hídrico antes da colheita / Free proline in Lippia sidoides Cham. under water stress before harvest

O estresse hídrico é considerado um dos fatores limitantes para o desenvolvimento das plantas, com isso, os vegetais desenvolvem mecanismos para tolerar a deficiência hídrica do solo. Entre esses mecanismos, está o acúmulo de prolina livre nos tecidos, tendo como função a regulação osmótica dos tecidos. A influência do estresse hídrico em plantas medicinais ainda é pouco estudado, principalmente em plantas nativas, como é o caso do alecrim-pimenta Lippia sidoides Cham. Assim, o trabalho teve por objetivo avaliar a prolina livre e o teor de flavonoides totais na referida espécie sob condições de estresse hídrico antes da colheita. O trabalho foi realizado no Instituto de Ciências Agrárias da UFMG, Campus Montes Claros, delineamento em blocos casualizados, com 5 tratamentos e 4 repetições, sendo eles 8, 6, 4, 2 e 0 dias de supressão da irrigação antes da colheita. Os teores encontrados de prolina variaram entre 0,2489 a 3,2421 ìM.g-1 de matéria seca e de flavonoides entre 0,0139 a 0,008 g.g-1. Os resultados observados indicam que a planta diminui seu teor de flavonoides e aumenta o teor de prolina com o aumento do período de estresse.

Water stress is considered a limiting factor for plant growth, thus, the plants develop mechanisms to tolerate the soil water deficit. Among these mechanisms is the accumulation of free proline in tissues, with the poke the osmotic regulation of tissues. The influence of water stress in medicinal plants is still poorly studied, mainly on native plants such as rosemary-pepper Lippia sidoides Cham. Thus, the study aimed to evaluate the free proline and total flavonoid content in that species under conditions of water stress before harvest. The study was conducted at the Institute of Agricultural Sciences, UFMG, Campus Montes Claros, randomized block design with 5 treatments and 4 replicates, and they 8, 6, 4, 2 and 0 days of withholding irrigation before harvest. The content of proline ranged from 0.2489 to 3.2421 ìM.g-1 of dry matter and flavonoids from 0.0139 to 0.008 gg-1. The results indicate that the plant reduces its content of flavonoids and proline content increases with increasing stress time.

Uso do propofol na indução anestésica de cutias (Dasyprocta sp.) / Use of propofol in the induction of anesthesia in agouti (Dasyprocta sp.)

Avaliou-se o emprego do propofol, por via intravenosa, como agente indutor anestésico em cutias (Dasyprocta sp), utilizando-se 10 fêmeas adultas, com pesos entre 1,5 a 2,0kg, criadas em cativeiro. Avaliaram-se as freqüências cardíaca e respiratória, a temperatura retal e a pressão arterial sistólica, antes da administração do propofol e logo após a anestesia dos animais. Foram mensurados o período hábil e de recuperação da anestesia. As médias das freqüências cardíaca e respiratória, temperatura retal e a pressão arterial sistólica basais foram de 220bcm, 92mrm, 38,95ºC e 110mmHg, respectivamente. Após a indução, as médias obtidas para essas variáveis foram de 214bcm, 84mrm, 39,05ºC e 80mmHg. As médias dos períodos hábil e de recuperação da anestesia foram de 10min e 45seg e 15min e 40seg, respectivamente. O propofol, quando utilizado em dose única, mostrou-se seguro, não alterando significativamente os parâmetros fisiológicos, podendo ser considerado um fármaco de escolha para a indução anestésica de cutias.

The endovenous use of propofol as inductor agent of anesthesia in agouti (Dasyprocta sp.) was evaluated using ten adut females, weighing 1.5 to 2.0kg, kept in captivity. Heart and respiratory rates, rectal temperature, and systolic arterial blood pressure were measured before and after anesthesia. Time of anesthesia and time of recovering of the anesthesia were also measured. The averages of the heart and respiratory rates, rectal temperature and the systolic blood pressure were 220bpm, 92mpm, 38.95ºC, and 110Hgmm, respectively. After the induction, the averages obtained for those variables were of 214bpm, 84mpm, 39.05ºC, and 80mmHg, respectively. The averages of the time of anesthesia and time of its recovery were 10min 45sec and 15min 40sec, respectively. Propofol when used in only one dose, was considered safe, since it did not disturbe the physiological parameters. Thus, propofol can be considered an alternative drug for anesthesia induction in agouti.

Investigation of human papillomavirus by hybrid capture II in cervical carcinomas including 113 adenocarcinomas and related lesions.

Hybrid capture is an easy and highly sensitive technique for screening population due to its capacity to detect malignant and premalignant lesions of the cervix. To evaluate its sensitivity, we investigated the frequency of high-risk human papillomavirus (HPV) infection and its correlation with glandular malignant lesions, analyzing a total of 113 cases of adenocarcinomas and related lesions. High-risk HPV was investigated using a hybrid capture II (HC2) assay. Samples were collected in two different ways: either brushed directly from surgical specimens before fixation or collected from the patients. We also investigated the frequency of HPV in squamous malignant lesions, 65 squamous cell carcinomas (SCC) and 66 in situ squamous cell carcinomas (ISSCC), to compare the occurrence of HPV for these lesions. The 113 glandular lesions comprised 62 invasive adenocarcinomas (IAC), 8 in situ adenocarcinomas (ISAC), 26 IAC plus SCC, and 17 adenosquamous cells carcinomas (ASCC). The HPV-positive reactions were as follows: 51 (82.2%) in IAC, 8 (100%) in ISAC, 25 (96.1%) in IAC plus SCC, and 14 (82.3%) in ASCC. HC2-positive results in the squamous malignant lesions were as follows: 58 of 63 (89.0%) for SCC and 94 of 103 (91.2%) for ISSCC. High-risk HPV infection was quite similar for glandular and pure squamous invasive malignant lesions, 82.2% and 89.0%, respectively, indicating that high-risk HPV is also highly prevalent in glandular lesions. Although hybrid capture proved to be an excellent adjunctive technique, we do not believe its results merit replacing the Pap smear as a screening tool.