Intoxicação por Metternichia princeps (Solanaceae) em caprinos no Estado da Bahia

Resumo:Descreve-se um surto de intoxicação por Metternichia princepsem caprinos no Estado da Bahia. De oito caprinos, três morreram, dos quais dois foram necropsiados; cinco não adoeceram. Os principais sinais clínicos caracterizaram-se por secreção nasal mucosa, emagrecimento, diarreia, apatia, debilidade leve, andar cambaleante, flexão dos membros torácicos e pélvicos, decúbito esterno-abdominal e decúbito lateral, seguidos de morte após aproximadamente dois dias de evolução clínica. Na necropsia foi observado edema pulmonar, hidrotórax, hidropericárdio, ascite, rins pálidos, edema perirrenal e hemorragias no tecido subcutâneo. Microscopicamente nos rins havia acentuada necrose de coagulação do epitélio tubular e túbulos com regeneração do epitélio. No pulmão havia acentuada congestão associada a edema interalveolar e interseptal. Na bioquímica sanguínea observou-se aumento na ureia, creatinina e creatinina fosfoquinase.

Co-infection by Tritrichomonas foetus and Pentatrichomonas hominis in asymptomatic cats

Abstract: Tritrichomonas foetus, a parasite well known for its significance as a venereally transmitted pathogen in cattle, has been identified as a cause of chronic large bowel diarrhea in domestic cats in many countries of the world. In Brazil, several studies on the diagnosis of bovine trichomoniasis have been performed, but until now, no study was made regarding feline trichomoniasis. Thus, this is the first study to report the occurrence of T. foetus and Pentatrichomonas hominis in cats using morphological and molecular analysis. Feces from 77 cats were examined, four of which (5.2%) were positive for the presence of parabasalids. Morphological analysis of stained smears revealed piriform trophozoites showing the three anterior flagella, elongated nucleus and axostyle ending abruptly in fillet, characteristic of T. foetus. In scanning and transmission electron microscopy, identification characters similar to those previously reported for T. foetus were observed. The cultures containing trophozoites were submitted for molecular analysis, which resulted positive for T. foetus DNA using specific primers (TFR3 and TFR4), and all samples were positive and subjected to sequencing in which they showed 99.7-100% similarity with another isolate sequencing of T. foetus (JX960422). Although no trophozoite with consistent morphology of P. hominis has been visualized in the samples, differential diagnosis was performed using specific primers for P. hominis (TH3 and TH5) amplicon. In three of the four samples (3.89%) sequencing revealed 100% similarity when compared with another sequence of P. hominis deposited in Genbank (KC623939). Therefore, the present study revealed through the diagnostic techniques employed the simultaneous infection by T. foetus and P. hominis in the feces of cats. However, it was necessary to use more than one technique for the diagnosis of the co-infection. These results demonstrate the importance of a correct diagnosis to allow an appropriate treatment by the veterinarian.

An experimental investigation of the effects of nozzle ellipticity on the flow structure of co-flow jet diffusion flames

The flow structure of cold and ignited jets issuing into a co-flowing air stream was experimentally studied using a laser Doppler velocimeter. Methane was employed as the jet fluid discharging from circular and elliptic nozzles with aspect ratios varying from 1.29 to 1.60. The diameter of the circular nozzle was 4.6 mm and the elliptic nozzles had approximately the same exit area as that of the circular nozzle. These non-circular nozzles were employed in order to increase the stability of attached jet diffusion flames. The time-averaged velocity and r.m.s. value of the velocity fluctuation in the streamwise and transverse directions were measured over the range of co-flowing stream velocities corresponding to different modes of flame blowout that are identified as either lifted or attached flames. On the basis of these measurements, attempts were made to explain the existence of an apparent optimum aspect ratio for the blowout of attached flames observed at higher values of co-flowing stream velocities. The insensitivity of the blowout limits of lifted flames to nozzle geometry observed in our previous work at low co-flowing stream velocities was also explained. Measurements of the fuel concentration at the jet centerline indicated that the mixing process was enhanced with the 1.38 aspect ratio jet compared with the 1.60 aspect ratio jet. On the basis of the obtained experimental data, it was suggested that the higher blowout limits of attached flames for an elliptic jet of 1.38 aspect ratio was due to higher entrainment rates.

Resistência de populações de capim-colchão (Digitaria ciliaris) aos herbicidas inibidores da Acetil Co-A Carboxilase

O objetivo do presente trabalho foi avaliar as populações com suspeita de ocorrência de biótipos resistentes de Digitaria ciliaris aos herbicidas inibidores da AcetilCoA Carboxilase (ACCase), por meio de curvas de dose-resposta, bem como estabelecer o grau de resistência cruzada aos herbicidas cicloexanodionas (CHD) e ariloxifenoxipropionatos (APP) desses biótipos. O experimento foi conduzido em casa de vegetação, utilizando-se quatro populações com suspeita de resistência (R1, R2, R3 e R4) e uma população suscetível (S). O delineamento experimental adotado foi o de blocos ao acaso, com quatro repetições, sendo os tratamentos resultado da interação fatorial entre cinco populações, três herbicidas (fluazifop-p-butil, sethoxydim e tepraloxydim) e oito doses de herbicidas (0C, 0,06C, 0,125C, 0,5C, 1C, 2C, 4C e 10C), em que C é a dose comercial recomendada para cada produto. Foram realizadas avaliações de porcentagem de controle aos 28 dias após a aplicação (DAA). A partir dos resultados obtidos, conclui-se que as populações R1, R2, R3 e R4 apresentaram-se como biótipos resistentes aos herbicidas inibidores da ACCase, com diferentes níveis de resistência cruzada aos herbicidas com esse mecanismo de ação.

Intoxicação de espécies de eucalipto submetidas à deriva do glyphosate

O glyphosate é o herbicida mais utilizado em áreas de reflorestamento de eucalipto. Nessas áreas tem sido freqüente a verificação de sintomas de intoxicação devido à deriva. Entretanto, trabalhos de pesquisa e observações de campo indicam comportamento diferencial entre as espécies e os clones de eucalipto quando em contato com o glyphosate. O presente trabalho teve como objetivo avaliar os efeitos da deriva simulada de glyphosate, por meio de doses reduzidas, no crescimento de cinco espécies de eucalipto. Utilizou-se o esquema fatorial 5x5, sendo cinco espécies (Eucalyptus urophylla, E. grandis, E. pellita, E. resinifera e E. saligna) e cinco doses (0; 43,2; 86,4; 172,8; e 345,6 g ha-1 de glyphosate), no delineamento em blocos casualizados com quatro repetições. A aplicação do herbicida foi feita sobre as plantas, de modo que não atingisse o terço superior, 23 dias após o plantio destas. Os sintomas de intoxicação causados pelo glyphosate foram semelhantes nas diferentes espécies, sendo caracterizados por murcha, clorose e enrolamento foliar, e, no caso de maiores doses, por necroses e senescência foliar. Plantas submetidas a doses acima de 86,4 g ha-1 de glyphosate foram severamente intoxicadas, afetando o seu crescimento, resultando em menor altura, diâmetro do caule e massa seca aos 45 dias após aplicação do herbicida. Entre as espécies estudadas, E. resinifera foi mais tolerante à deriva de glyphosate, apresentando menores valores de intoxicação e maior incremento em altura e diâmetro, mesmo nas plantas submetidas às maiores doses, o que não foi observado nas demais espécies.

Intoxicação de eucalipto submetido à deriva simulada de diferentes herbicidas

Além do glyphosate, amplamente usado, outros herbicidas, como o triclopyr e o carfentrazone-ethyl, apresentam potencial de uso na eucaliptocultura. Entretanto, a nãoseletividade destes herbicidas ao eucalipto pode levar à intoxicação da cultura, caso haja contato das folhas com a calda aplicada. Objetivou-se avaliar os efeitos da deriva de alguns herbicidas em plantas de Eucalyptus urophylla, simulada por meio de subdoses: 0, 43,2, 86,4 e 172,8 g ha-1 de glyphosate; 14,4, 28,8 e 57,6 g ha-1 de triclopyr; 0,84, 1,68 e 3,36 g ha-1 de carfentrazone-ethyl; e das misturas: 43,2 g ha-1 de glyphosate + 14,4 g ha-1 de triclopyr; 86,4 g ha¹ de glyphosate + 28,6 g ha-1 de triclopyr; 43,2 g ha-1 de glyphosate + 1,68 g ha-1 de carfentrazone-ethyl; 86,4 g ha-1 de glyphosate + 3,36 g ha-1 de carfentrazone-ethyl. As mudas de eucalipto com aproximadamente 50 cm de altura receberam a aplicação dos herbicidas de modo a não atingir o terço superior das plantas, 45 dias após o transplantio. Entre os tratamentos, a deriva de glyphosate mostrou-se mais danosa ao eucalipto, seguida de carfentrazone-ethyl e triclopyr. Plantas de eucalipto expostas à subdose de 172,8 g ha-1 de glyphosate e às misturas glyphosate + carfentrazone-ethyl tiveram maior porcentagem de intoxicação e menor crescimento aos 15, 30 e 45 dias após aplicação (DAA). Houve recuperação das plantas expostas à deriva do triclopyr, que apresentaram 56,25 e 15,10% de intoxicação aos 15 e 30 DAA, respectivamente, indicando menor risco da deriva do triclopyr em comparação aos demais herbicidas e misturas estudados.

Avaliação da intoxicação de cultivares de cana-de-açúcar e I. grandifolia ao amicarbazone

O objetivo deste trabalho foi avaliar a intoxicação de planta daninha e cultivares de cana-de-açúcar ao amicarbazone. Para isso, utilizou-se Ipomoea grandifolia como planta daninha representante e os cultivares de cana-de-açúcar PO8862, SP80 3280 e RB83 5486, caracterizados como sensível, intermediário e tolerante aos herbicidas, respectivamente. Foi verificado o consumo de água e quantificada a concentração do amicarbazone em seiva de xilema dos três cultivares de cana-de-açúcar e de I. grandifolia por meio da bomba de Schollander e da cromatografia e espectrometria de massas (LC-MS). A intoxicação das plantas foi verificada através de leituras da fluorescência, com auxílio do fluorômetro portátil, que permitiu a correlação da taxa de transporte de elétrons (ETR) com a concentração de amicarbazone absorvido pelos cultivares de cana-de-açúcar e por I. grandifolia. Verificou-se, através do experimento, que a redução dos valores da ETR pode ser utilizada para indicar o nível de intoxicação de I. grandifolia e de plantas de cana-de-açúcar ao amicarbazone. I. grandifolia destacou-se em relação à cana-de-açúcar pela maior sensibilidade ao amicarbazone. A suscetibilidade diferencial dos cultivares de cana-de-açúcar PO8862, SP80 3280 e RB83 5486 pode ser justificada, possivelmente, pela absorção diferencial do amicarbazone entre os cultivares.

Sintomas da intoxicação inicial de Eucalyptus proporcionados por subdoses de glyphosate aplicadas no caule ou nas folhas

Com o objetivo de avaliar os sintomas de intoxicação causados pela aplicação de glyphosate, foi montado um estudo composto por quatro ensaios com aplicações de glyphosate (360 g e.a. L-1) em eucalipto. Em todos os ensaios, mudas foram transplantadas em vasos de 5,0 L. Nos ensaios 1 e 2, foram aplicados volumes crescentes de solução de glyphosate no caule do eucalipto. No ensaio 1, a solução de 3% (v/v) foi aplicada nos volumes de 0, 1, 5, 10, 20, 40, 80 e 160 µL de calda por planta e, no segundo, a solução de glyphosate a 2% (v/v) foi aplicada nos volumes de 0, 1, 5, 15, 30, 60, 90, 120 e 150 µL de calda por planta. Nos ensaios 3 e 4, foram feitas aplicações de glyphosate sobre as plantas de eucalipto. No ensaio 3, as doses foram de 0, 7,2.10-7, 7,2.10-6, 7,2.10-5, 7,2.10-4, 7,2.10-3, 7,2.10-2, 7,2.10-1, 7,2, 72, 360 e 720 g e.a. de glyphosate ha-1 e, no ensaio 4, de 0, 9, 18, 36, 72, 144, 288, 432, 576, 720, 1.080, 1.440 e 2.160 g e.a. de glyphosate ha-1. Nos quatro ensaios foi utilizado o delineamento DIC, com três repetições. Nas plantas, foram avaliadas a altura, a área foliar e a matéria seca de caule e folhas. Os resultados obtidos foram submetidos a análises de regressão. Quando aplicadas no caule, doses de 40,78 e 51,41 µL de calda por planta de glyphosate a 3 e 2% (v/v), respectivamente, nos ensaios 1 e 2, foram suficientes para redução média de 50% das características estudadas. Nas aplicações sobre as folhas, houve maior sensibilidade das plantas mais desenvolvidas. Para redução média de 50% nas variáveis analisadas, foram necessárias doses de 277,4 e 143,3 g e.a. de glyphosate ha-1 nos ensaios 3 e 4, respectivamente.

Poly-DL-lactide-co-glycolide microspheres as a controlled release antigen delivery system

Successful vaccine application means maximum protection with minimal number of administrations. A rational development of vaccines involves studies of the nature of the antigen as well as of the adjuvant to be used to improve the immune responses. This has provided the impetus for studies to design the degradable devices and for different approaches to antigen delivery by different routes of administration. The development of controlled release systems based on polymeric devices that permit a sustained or pulsed release of encapsulated antigens has attracted much interest. Polymeric delivery systems consist of polymers that release their content continuously in a controlled manner over a period of time. The development of a biocompatible delivery system for parenteral administration offers several advantages in terms of immunoadjuvanticity over other compounds. It was found that, in contrast to other carriers, microspheres are more stable, thus permitting administration by the oral or parenteral route. In the present study, we describe the main characteristics and potentialities of this new immunoadjuvant for oral and parenteral administration.

Modulation of intercellular communication by differential regulation and heteromeric mixing of co-expressed connexins

Intercellular communication may be regulated by the differential expression of subunit gap junction proteins (connexins) which form channels with differing gating and permeability properties. Endothelial cells express three different connexins (connexin37, connexin40, and connexin43) in vivo. To study the differential regulation of expression and synthesis of connexin37 and connexin43, we used cultured bovine aortic endothelial cells which contain these two connexins in vitro. RNA blots demonstrated discordant expression of these two connexins during growth to confluency. RNA blots and immunoblots showed that levels of these connexins were modulated by treatment of cultures with transforming growth factor-ß1. To examine the potential ability of these connexins to form heteromeric channels (containing different connexins within the same hemi-channel), we stably transfected connexin43-containing normal rat kidney (NRK) cells with connexin37 or connexin40. In the transfected cells, both connexin proteins were abundantly produced and localized in identical distributions as detected by immunofluorescence. Double whole-cell patch-clamp studies showed that co-expressing cells exhibited unitary channel conductances and gating characteristics that could not be explained by hemi-channels formed of either connexin alone. These observations suggest that these connexins can readily mix with connexin43 to form heteromeric channels and that the intercellular communication between cells is determined not only by the properties of individual connexins, but also by the interactions of those connexins to form heteromeric channels with novel properties. Furthermore, modulation of levels of the co-expressed connexins during cell proliferation or by cytokines may alter the relative abundance of different heteromeric combinations.

A third generation regimen VACOP-B with or without adjuvant radiotherapy for aggressive localized non-Hodgkin's lymphoma: report from the Italian Non-Hodgkin's Lymphoma Co-operative Study Group

The objective of this multicenter prospective study was to determine the clinical efficacy and toxicity of a polychemotherapeutic third generation regimen, VACOP-B, with or without radiotherapy as front-line therapy in aggressive localized non-Hodgkin's lymphoma. Ninety-three adult patients (47 males and 46 females, median age 45 years) with aggressive localized non-Hodgkin's lymphoma, 43 in stage I and 50 in stage II (non-bulky), were included in the study. Stage I patients received VACOP-B for 6 weeks plus involved field radiotherapy and stage II patients received 12 weeks VACOP-B plus involved field radiotherapy on residual masses. Eighty-six (92.5%) achieved complete remission and 4 (4.3%) partial remission. Three patients (3.2%) were primarily resistant. Ten-year probability of survival, progression-free survival and disease-free survival were 87.3, 79.9 and 83.9%, respectively. Eighty-four patients are surviving at a median observation time of 57 months (range: 6-126). Statistical analysis showed no difference between stages I and II in terms of response, ten-year probability of survival, progression-free survival or disease-free survival. Side effects and toxicity were negligible and were similar in the two patient groups. The results of this prospective study suggest that 6 weeks of VACOP-B treatment plus radiotherapy may be the therapy of choice in stage I aggressive non-Hodgkin's lymphoma. Twelve weeks of VACOP-B treatment with or without radiotherapy was shown to be effective and feasible for stage II. These observations need to be confirmed by a phase III study comparing first and third generation protocols in stage I-II aggressive non-Hodgkin's lymphoma.

Pressure-assisted cold denaturation of hen egg white lysozyme: the influence of co-solvents probed by hydrogen exchange nuclear magnetic resonance

COSY proton nuclear magnetic resonance was used to measure the exchange rates of amide protons of hen egg white lysozyme (HEWL) in the pressure-assisted cold-denatured state and in the heat-denatured state. After dissolving lysozyme in deuterium oxide buffer, labile protons exchange for deuterons in such a way that exposed protons are substituted rapidly, whereas "protected" protons within structured parts of the protein are substituted slowly. The exchange rates k obs were determined for HEWL under heat treatment (80ºC) and under high pressure conditions at low temperature (3.75 kbar, -13ºC). Moreover, the influence of co-solvents (sorbitol, urea) on the exchange rate was examined under pressure-assisted cold denaturation conditions, and the corresponding protection factors, P, were determined. The exchange kinetics upon heat treatment was found to be a two-step process with initial slow exchange followed by a fast one, showing residual protection in the slow-exchange state and P-factors in the random-coil-like range for the final temperature-denatured state. Addition of sorbitol (500 mM) led to an increase of P-factors for the pressure-assisted cold denatured state, but not for the heat-denatured state. The presence of 2 M urea resulted in a drastic decrease of the P-factors of the pressure-assisted cold denatured state. For both types of co-solvents, the effect they exert appears to be cooperative, i.e., no particular regions within the protein can be identified with significantly diverse changes of P-factors.

Use of blends of bioabsorbable poly(L-lactic acid)/poly(hydroxybutyrate- co-hydroxyvalerate) as surfaces for Vero cell culture

Vero cells, a cell line established from the kidney of the African green monkey (Cercopithecus aethiops), were cultured in F-10 Ham medium supplemented with 10% fetal calf serum at 37°C on membranes of poly(L-lactic acid) (PLLA), poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and their blends in different proportions (100/0, 60/40, 50/50, 40/60, and 0/100). The present study evaluated morphology of cells grown on different polymeric substrates after 24 h of culture by scanning electron microscopy. Cell adhesion was also analyzed after 2 h of inoculation. For cell growth evaluation, the cells were maintained in culture for 48, 120, 240, and 360 h. For cytochemical study, the cells were cultured for 120 or 240 h, fixed, processed for histological analysis, and stained with Toluidine blue, pH 4.0, and Xylidine ponceau, pH 2.5. Our results showed that cell adhesion was better when 60/40 and 50/50 blends were used although cells were able to grow and proliferate on all blends tested. When using PLLA/PHBV (50/50) slightly flattened cells were observed on porous and smooth areas. PLLA/PHBV (40/60) blends presented flattened cells on smooth areas. PLLA/PHBV (0/100), which presented no pores, also supported spreading cells interconnected by thin filaments. Histological sections showed that cells grew as a confluent monolayer on different substrates. Cytochemical analysis showed basophilic cells, indicating a large amount of RNA and proteins. Hence, we detected changes in cell morphology induced by alterations in blend proportions. This suggests that the cells changed their differentiation pattern when on various PLLA/PHBV blend surfaces.

Pim-1 kinase inhibits the activation of reporter gene expression in Elk-1 and c-Fos reporting systems but not the endogenous gene expression: an artifact of the reporter gene assay by transient co-transfection

We have studied the molecular mechanism and signal transduction of pim-1, an oncogene encoding a serine-threonine kinase. This is a true oncogene which prolongs survival and inhibits apoptosis of hematopoietic cells. In order to determine whether the effects of Pim-1 occur by regulation of the mitogen-activated protein kinase pathway, we used a transcriptional reporter assay by transient co-transfection as a screening method. In this study, we found that Pim-1 inhibited the Elk-1 and NFkappaB transcriptional activities induced by activation of the mitogen-activated protein kinase cascade in reporter gene assays. However, Western blots showed that the induction of Elk-1-regulated expression of endogenous c-Fos was not affected by Pim-1. The phosphorylation and activation of neither Erk1/2 nor Elk-1 was influenced by Pim-1. Also, in the gel shift assay, the pattern of endogenous NFkappaB binding to its probe was not changed in any manner by Pim-1. These data indicate that Pim-1 does not regulate the activation of Erk1/2, Elk-1 or NFkappaB. These contrasting results suggest a pitfall of the transient co-transfection reporter assay in analyzing the regulation of transcription factors outside of the chromosome context. It ensures that results from reporter gene expression assay should be verified by study of endogenous gene expression.