An open-label randomized clinical trial to evaluate the efficacy of everolimus versus tacrolimus in triple maintenance immunosuppressive therapy for kidney transplant patients.

Tacrolimus (TAC), a calcineurin inhibitor, and everolimus (EVL), an mTOR inhibitor, have been used as immunosuppressive (ISS) drugs in post-kidney transplantation therapy. The objective of this study was to compare the efficacy of EVL vs TAC in the ISS maintenance triple therapy. Ninety-seven kidney transplant patients, who received triple maintenance therapy with TAC, mycophenolate mofetil (MMF), and methyl prednisone (PRED), were evaluated. After four months of post-kidney transplant therapy, 30 patients enrolled in a randomized controlled clinical trial, in which 16 patients received TAC+MMF+PRED (cohort 1), and 14 patients switched to EVL+MMF+PRED (cohort 2). The patients were followed-up for 36 months. Two patients from cohort 1 lost their grafts after one year due to non-adherence. Two patients from cohort 2 had intolerance to mTOR inhibitors and were switched back to TAC from EVL. One case (6.25%) in cohort 1 and three cases (21.43%) in cohort 2 of acute T-cell-mediated rejection was observed. Antibody-mediated acute rejection (ABMAR) was observed in four patients (25.0%) in cohort 1, and antibody-mediated chronic rejection (ABMCR) was observed in two patients (12.50%). One patient from cohort 2 lost the graft after 15 months due to polyomavirus infection. The graft survival rate was 87.50% in cohort 1 and 92.86% in cohort 2. This clinical trial showed that the EVL+MMF+PRED triple maintenance therapy was efficacious compared with TAC during 32 months of follow-up. However, further studies are needed to confirm the efficacy of this regimen for long-term graft survival.

An open-label randomized clinical trial to evaluate the efficacy of everolimus versus tacrolimus in triple maintenance immunosuppressive therapy for kidney transplant patients

Tacrolimus (TAC), a calcineurin inhibitor, and everolimus (EVL), an mTOR inhibitor, have been used as immunosuppressive (ISS) drugs in post-kidney transplantation therapy. The objective of this study was to compare the efficacy of EVL vs TAC in the ISS maintenance triple therapy. Ninety-seven kidney transplant patients, who received triple maintenance therapy with TAC, mycophenolate mofetil (MMF), and methyl prednisone (PRED), were evaluated. After four months of post-kidney transplant therapy, 30 patients enrolled in a randomized controlled clinical trial, in which 16 patients received TAC+MMF+PRED (cohort 1), and 14 patients switched to EVL+MMF+PRED (cohort 2). The patients were followed-up for 36 months. Two patients from cohort 1 lost their grafts after one year due to non-adherence. Two patients from cohort 2 had intolerance to mTOR inhibitors and were switched back to TAC from EVL. One case (6.25%) in cohort 1 and three cases (21.43%) in cohort 2 of acute T-cell-mediated rejection was observed. Antibody-mediated acute rejection (ABMAR) was observed in four patients (25.0%) in cohort 1, and antibody-mediated chronic rejection (ABMCR) was observed in two patients (12.50%). One patient from cohort 2 lost the graft after 15 months due to polyomavirus infection. The graft survival rate was 87.50% in cohort 1 and 92.86% in cohort 2. This clinical trial showed that the EVL+MMF+PRED triple maintenance therapy was efficacious compared with TAC during 32 months of follow-up. However, further studies are needed to confirm the efficacy of this regimen for long-term graft survival.

Animal performance and nutritional characteristics of Piatã-grass in integrated systems / Desempenho animal e características nutricionais de capim-piatã em sistemas integrados

The objective was to evaluate animal performance and nutritional characteristics of Brachiaria brizantha cv. BRS Piatã in two integrated systems, during the summer and winter, five years after the initial establishment of the area. The randomized block experimental design was adopted with treatments consisting of a control (five native trees per hectare) and an integrated crop-livestock-forestry (ICLF) system with 227 eucalyptus trees per hectare. Animal performance, canopy height, soil cover, forage mass (leaf, stem and senescent material), and forage nutritive value were evaluated. Differences between systems were observed in forage mass (total, leaf, stem and senescent material), soil cover and forage nutritive value in both seasons. Shading conditions provided by eucalyptus in the ICLF system lead to a reduction in forage mass and neutral detergent fiber content, and to an increase in crude protein and in vitro organic matter digestibility. However, no effect on animal performance was obtained. The presence of trees improves the forage nutritive value without impacting animal performance in integrated systems.(AU)

O objetivo foi avaliar o desempenho animal e as características nutricionais de Brachiaria brizantha cv. BRS Piatã em dois sistemas integrados, durante o verão e o inverno, cinco anos após o estabelecimento inicial da área. O delineamento experimental em blocos ao acaso foi adotado com tratamentos constituídos por um controle (cinco árvores nativas por hectare) e um sistema integrado lavoura-pecuária-floresta (ILPF) com 227 eucaliptos por hectare. Desempenho animal, altura do dossel, cobertura do solo, massa de forragem (folha, caule e material senescente) e valor nutritivo da forragem foram avaliados. Diferenças entre os sistemas foram observadas na massa de forragem (total, foliar, caule e material senescente), na cobertura do solo e no valor nutritivo da forragem em ambas as estações. As condições de sombreamento proporcionadas pelo eucalipto no sistema ICLF levaram a uma redução na massa de forragem e no teor de fibra em detergente neutro e a um aumento na proteína bruta e na digestibilidade in vitro da matéria orgânica. No entanto, nenhum efeito no desempenho animal foi obtido. A presença de árvores melhora o valor nutritivo da forragem, sem impactar o desempenho animal em sistemas integrados.(AU)

Massa de forragem, composição morfológica e valor nutritivo de capim-braquiária submetido a níveis de sombreamento e fertilização / Herbage mass, morphological composition and nutritive value of signalgrass, submitted to shading and fertilization levels

Características produtivas e nutricionais de Brachiaria decumbens, submetida a condições de sombreamento (sol pleno, 20% e 70% de sombreamento), sem e com uso de fertilização (80 kg.ha-1 de N e K2O e 60 kg.ha-1 de P2O5), foram avaliadas no delineamento de blocos casualizados, com quatro repetições. As parcelas sombreadas foram alocadas em um sistema silvipastoril, enquanto aquelas a pleno sol foram avaliadas em um monocultivo de B. decumbens. O sombreamento severo promoveu reduções de 54% na massa seca verde, 59% na massa seca total e 58% na densidade volumétrica de forragem. Aumentos da massa de forragem e da densidade volumétrica em resposta ao fertilizante foram observados somente nas condições de sol pleno e sombreamento moderado. A adubação não influenciou nas características nutricionais da forragem, mas o sombreamento severo promoveu aumentos de 20 e 51% nos teores de clorofila e de proteína bruta, respectivamente. Reduções dos teores de fibra em detergente neutro e fibra em detergente ácido foram observadas com o sombreamento. Os teores de lignina e a digestibilidade in vitro da matéria seca não variaram com nenhum fator estudado (médias de 6,0 e 60,9%, respectivamente). A adubação de B. decumbens com dose moderada de NPK é recomendada para dosséis cultivados no sol pleno e sombreamento moderado, mas deve ser evitada em pastos severamente sombreados, pois não reflete aumento de massa de forragem nem melhorias de seu valor nutricional.(AU)

Productive and nutritional traits of Brachiaria decumbens, submitted to shading conditions (full sun, 20% and 70% shading) and fertilization levels (without or with 80 kg.ha-1 of N and K 2 O and 60 kg.ha-1 of P 2O5), were evaluated in a randomized complete block design, with four replications. The shading were allocated in a silvopastoral system, while full sun was obtained in an exclusive B. decumbens pasture. The severe shading reduced the green dry mass in 54%, total dry mass in 59% and the forage bulk density in 58%. Increases in the forage mass and forage bulk density in response to fertilization were observed only in full sun and moderate shade. The nutritional traits did not vary with fertilization, but the chlorophyll and crude protein contents increased 20 and 51% with severe shading, respectively. The neutral detergent fiber and acid detergent fiber reduced with shade. The lignin content and in vitro dry matter digestibility did not vary with factors (average values of 6.0 and 60.9%, respectively). Fertilization with moderate dose of NPK is recommended to B. decumbens in full sun and moderate shading, but should be avoided in severely shaded pasture, because not reflected in improvement of forage mass and its nutritional traits.(AU)

Combined use of Paracoccidioides brasiliensis recombinant rPb27 and rPb40 antigens in an enzyme-linked immunosorbent assay for immunodiagnosis of paracoccidioidomycosis.

Paracoccidioidomycosis (PCM) is one of the most important endemic mycoses in Latin America; it's usually diagnosed by observation and/or isolation of the etiologic agent, Paracoccidioides brasiliensis, as well as by a variety of immunological methods, such as complement fixation and immunodiffusion. Although these approaches are useful, historically their sensitivity and specificity have often been compromised by the use of complex mixtures of undefined antigens. The use of combinations of purified, well-characterized antigens appears preferable and may yield good results. In the present study combinations of the previously described 27-kDa recombinant antigen (rPb27) and a recombinant 40-kDa-molecular-mass antigen (rPb40) from this fungus, that was identified by Goes et al. (2005) through the AST strategy as a homolog of Neurospora crassa calcineurin B, were used in an indirect enzyme linked immunosorbent assay (ELISA) for diagnosis and follow-up of patients with PCM. The complete coding cDNA of rPb40 and rPb27 were cloned into a pET-21a and a pET-DEST 42 plasmid, respectively, expressed in E. coli with a his-tag and purified by affinity chromatography. Among 109 PCM serum samples analyzed, a homogeneous IgG response to these proteins was observed. 62 serum samples from patients with other diseases, 18 from patients with other mycosis and 23 from healthy individuals were also studied. Detection of anti-rPb27 and anti-rPb40 antibodies in sera of patients with PCM by ELISA using a combination of the two purified proteins showed a sensitivity of 96% with a specificity of 100% in relation to control normal human sera and to sera from patients with other systemic mycosis and 93.5% to sera from patients with diverse infections. The use of this two proteins combination provided an excellent immunodiagnosis assay with great values of sensitivity and specificity, even in relation to sera from patients with other mycosis, making possible the standardization of a new methodology to diagnose this important mycosis, with a good confiability and reprodutibility.

Avaliação imunohistoquímica e ultraestrutural de gametas e embriões caprinos infectados com o CAEV / Immunohistochemical and ultrastructural evaluation of gametes and embryos of goats infected with CAEV

O objetivo do presente estudo foi determinar a susceptibilidade dos folículos ovarianos, espermatozoides e embriões caprinos ao Vírus da Artrite Encefalite Caprina (CAEV). Para isto, foram analisados espermatozoides e folículos ovarianos pelas técnicas de imunohistoquímica e microscopia eletrônica de transmissão, antes e após protocolos de infecção in vitro com o CAEV. Foram submetidos à análise ultraestrutural, embriões caprinos produzidos in vivo, oriundos de cabras negativas e positivas para o CAEV. Nas amostras seminais, provenientes de animais tanto com infecção natural quanto dos artificialmente infectados, foi observada imunomarcação positiva dos espermatozoides, assim como alterações degenerativas na sua análise ultraestrutural. Já nas amostras de tecido ovariano, a imunomarcação foi mais discreta e identificada na região do estroma. No tocante à análise ultraestrutural, folículos e embriões se apresentaram íntegros. De acordo com esses resultados, pode-se concluir que os espermatozoides caprinos apresentaramse infectados, assinalando a susceptibilidade dessas células ao vírus, bem como a potencialidade do CAEV ser carreado ao cerne do oócito, originando embriões infectados.

The aim of this study was to determine the susceptibility of goat ovarian follicles, spermatozoa and embryos to caprine arthritis-encephalitis virus (CAEV). Spermatozoa and ovarian follicles were analyzed, before and after in vitro infection with CAEV, through immunohistochemistry and transmission electron microscopy techniques. Goat embryos, produced in vivo from infected and non-infected goats, were submitted to ultrastructural analysis. Immunohistochemical examination of seminal samples from goats naturally and artificially infected with CAEV revealed viral antigens in spermatozoa, while the ultrastructural analysis showed degenerative changes in these cells. Ovarian tissue samples presented a more discreet immunohistochemical positive reaction situated in the stroma region. Ultrastructural analysis revealed that the embryos and ovarian follicles were intact. These results indicate that the spermatozoa were infected, confirming the susceptibility of these cells to the virus, as well as the potential of CAEV entering the oocyte, giving rise to infected embryos.

Inibição dos lentivírus de pequenos ruminantes por drogas antivirais / Inhibition of small ruminant lentivirus by antiviral drugs

Inibidores da enzima transcriptase reversa e da protease foram avaliados quanto ao seu efeito inibitório na replicação do Vírus da Artrite Encefalite Caprina (CAEV) cepa CAEV Cork e do vírus Maedi-Visna (MVV) cepa K1514 cultivados em células fibroblásticas de caprinos. Os fármacos utilizados foram: lamivudina, didanosina, estavudina, zidovudina, efavirenz, atazanavir e lopinavir/ritonavir. A maior concentração utilizada para lamivudina, estavudina, zidovudina e efavirenz foi 500 ?M, para atazanavir foi 50 ?M e 5,0 ?M para lopinavir/r e didanosina. A atividade antiviral in vitro foi pesquisada por meio da avaliação da viabilidade celular através da redução do MTT e pela pesquisa de inibição dos efeitos citopáticos (CPE) dos vírus. A replicação dos vírus só não foi completamente bloqueada pelos inibidores de protease (IP) atazanavir e lopinavir/r enquanto os demais apresentaram eficácia antiviral significativa em diferentes concentrações. Os IP juntamente com o efavirenz, não mostraram atividade antiviral quando foram avaliados pela técnica de redução do MTT. Esses dados indicam que os fármacos inibidores da transcriptase reversa lamivudina, didanosina, estavudina e zidovudina são eficazes na inibição in vitro dos lentivírus de pequenos ruminantes.

Inhibitors of the reverse transcriptase and protease enzymes were evaluated for their inhibitory effect on the replication of caprine arthritis encephalitis virus (CAEV) strain CAEV Cork and of the Maedi-Visna virus (MVV) strain K1514 cultured in fibroblastic cells. The drugs lamivudine, didanosine, stavudine, zidovudine, efavirenz, atazanavir and lopinavir/ritonavir were used. The highest concentration used for lamivudine, stavudine, zidovudine and efavirenz was 500 ?M, for atazanavir it was 50 ?M and 5.0 ?M for lopinavir/r and didanosine. The in vitro antiviral activity was investigated by evaluating the cell viability by the MTT method and testing for inhibition of cytopathic effects (CPE) of the virus. The replication of the virus was not completely inhibited by the protease inhibitors atazanavir and lopinavir/r in the test for CPE, while the others drugs showed significant antiviral efficacy in different concentrations. The protease inhibitors together with the efavirenz did not show antiviral activity when they were assessed by the reduced MTT technique. These data showed that the reverse transcriptase inhibitor drugs lamivudine, didanosine, stavudine and zidovudine were effective in the in vitro inhibition of small ruminant lentivirus.

Avaliaçäo auditiva em escolares / Hearing evaluation of school children

Introduçäo: A avaliaçäo auditiva na faixa etária escolar é necessária para a identificaçäo e correçäo precoce das alteraçöes auditivas. Objetivos: Identificar e quantificar as alteraçöes audiométricas mais freqüentes em escolares. Casuística e Método: Pesquisa realizada com 121 escolares da 1ª a 8ª séries, faixa etária de 7 a 14 anos no mês de novembro de 1998 na cidade de Goiânia, por médicos otorrinolaringologistas e fonoaudiólogos através de exame clínico otorrinolaringológico, audiometria tonal e imitânciometria no Ambulatório de Otorrinolaringologia do Hospital das Clínicas da Faculdade de Medicina da Universidade Federal de Goiás. Resultados: Audiometrias realizadas em 242 orelhas foram encontradas: 76 por cento (184) orelhas com audiometria dentro dos limites da normalidade e 24 por cento (58) orelhas com audiometria alterada. As alteraçöes audiométricas mais freqüentes foram: perda auditiva condutiva em 12 por cento (26) sendo 8 por cento (16) esquerdas e 4 por cento (9) direitas, perda auditiva neurossensorial leve em 7 por cento (15) orelhas sendo 2 por cento (5) esquerdas e 5 por cento (10) direitas. Imitânciometria realizada em 242 orelhas obtendo timpanometrias alteradas sendo: curva do tipo B 3 por cento (6) e do tipo C 3 por cento (6). Conclusäo: Concluímos que as alteraçöes auditivas que ocorrem podem prejudicar a atençäo e compreensäo dos escolares, sendo importante o seu diagnóstico e tratamento precoce para obtençäo de um melhor rendimento escolar

Epistaxe como manifestaçäo freqüente da síndrome de Bernard-Soulier / Epistaxis as frequent manifestation of Bernard-Soulier syndrome

Os autores relatam um caso de Síndrome de Bernard-Soulier atendido na unidade de emergência do Hospital das Clínicas da Faculdade de Medicina da Universidade Federal de Goiás com manifestaçöes nasais, orais e gastrintestinais. Discutem consideraçöes sobre o quadro clínico, diagnóstico, tratamento e evoluçäo da doença. Alertam sobre a importância de considerar a possibilidade de discrasias sangüíneas quando frente a epistaxes de repetiçäo e/ ou de difícil tratamento

Genetic analysis of the bacteriocin-encoding plasmids pRJ6 and pRJ9 of Staphylococcus aureus by transposon mutagenesis and cloning of genes involved in bacteriocin production.

pRJ6 and pRJ9, small Staphylococcus aureus plasmids which code for bacteriocins, exhibited a bactericidal activity against several lactic acid bacteria and strains of Listeria monocytogenes, an important food-borne pathogen. Filter-mating experiments using plasmid derivatives tagged with either Tn551 or Tn917-lac showed that pRJ6, but not pRJ9, could be mobilized by staphylococcal conjugative plasmids. Transposon mutagenesis of both plasmids was also performed. The bacteriocin and immunity structural genes of pRJ6 are part of the same operon, which is located around co-ordinate 4.0, being transcribed from right to left. However, gene cloning experiments using a staphylococcal vector showed some evidence for the involvement of additional functions of pRJ6 in bacteriocin expression. One function involved in pRJ6 mobilization mapped around co-ordinate 5.2, and it appears to be transcribed from left to right. The bactericidal action exerted by strains harbouring pRJ9 appears to reflect the activity of at least two bacteriocins, whose combined action results in a broader spectrum of activity and in a higher antagonistic activity. Gene cloning experiments also supported these assumptions.

Incompatibility and molecular relationships between small bacteriocinogenic plasmids of Staphylococcus aureus.

The sequence relations between small bacteriocinogenic plasmids (pRJ6, pRJ9, pRJ10 and pRJ11) of Staphylococcus aureus were investigated by comparing restriction maps and by hybridization. Plasmids pRJ10 and pRJ11 showed identical restriction maps, similar to that of pRJ9. The restriction map of pRJ6 differed from those of pRJ9 and pRJ10/pRJ11. Both groups of plasmids were shown to share a region of homology of at least 2.6 kb. The incompatibility relationships between them were also investigated by using plasmid derivatives tagged with transposon Tn551. Plasmids pRJ6 and pRJ9 proved to belong to different incompatibility groups.

Cloning of the replication region on the bacteriocinogenic plasmid pRJ9 of Staphylococcus aureus.

A 5.8-kb ClaI fragment of pRJ9, a bacteriocinogenic plasmid of Staphylococcus aureus, was cloned in the unique ClaI site of pRJ5. The recombinant plasmid obtained, pRJ23, failed to confer bacteriocin production and immunity to bacteriocin on host cells. The cloned fragment was shown to contain the complete replicon of pRJ9. Attempts to clone the 4.4-kb ClaI fragment of pRJ9 were unsuccessful, apparently due to the inactivation of the basic replicon of the cloning vector. Therefore, plasmid pRJ5 cut at its ClaI site appears to be a suitable vector for cloning replication regions of plasmids that can replicate in S. aureus.