Avaliação bioquímica e hostológica de fígado de ratas wistar diabéticas e tratadas com tamoxifeno

Tamoxifen (TX), a drug used in the treatment of breast cancer, may cause hepatic changes in some patients. The consequences of its use on the liver tissues of rats with or without diabetes mellitus (DM) have not been fully explored. The purpose of this multidisciplinary study was to evaluate the correlation between plasma hepatic enzyme levels and the presence of iron overload in the hepatic tissue of female Wistar rats with or without streptozotocin-induced DM and using TX. Female rats were studied in control groups: C-0 (non-drug users), C-V (sorbitol vehicle only) and C-TX (using TX). DM (diabetic non-drug users) and DM-TX (diabetics using TX) were the test groups. Sixty days after induced DM, blood samples were collected for glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST) alkaline phosphatase (ALP) and bilirubin measures. Hepatic fragments were processed and stained with hematoxylin and eosin (H&E), Masson s trichrome, Perls. The hepatic iron content was quantified by atomic absorption spectrometry. AST, ALT and ALP levels were significantly elevated in the DM and DM-TX groups, with unchanged bilirubin levels. Liver iron overload using Perls stain and atomic absorption spectrometry were observed exclusively in groups C-TX and DM-TX. There was positive correlation between AST, ALT and ALP levels and microscopic hepatic siderosis intensity in group DM-TX. In conclusion, TX administration is associated with liver siderosis in diabetic and non-diabetic rats. In addition, TX induced liver iron overload with unaltered hepatic function in 2 non-diabetic rats and may be a useful tool for investigating the biological control of iron metabolism

Emprego da citometria de fluxo na avaliação do perfil imunofenotípico de pacientes com leucemia linfocítica crônica

Chronic lymphocytic leukemia (B-CLL) is a clonal proliferation of mature B lymphocytes characterized by indolent clinical course. Biologically this clonallity is characterized by low expression of surface immunoglobulin (sIg) with restriction to a single immunoglobulin light chain associated with high expression of CD5 antigen and positivity to B cell antigens lymphocytes such as CD19, CD20 and CD23 and negativity to FMC7. The immunological profile and morphological analysis of lymphoid cells are the main means for the differential diagnosis of B-CLL from other chronic lymphoproliferative diseases. The aim of this study was to evaluate the expression pattern of a variety of membrane antigens in leukemic cells originating from patients with B-CLL. In this study, peripheral blood samples from 80 patients with B-CLL were analyzed by multiparametric flow cytometry in addition to routine hematologic exams, using a panel of monoclonal antibodies (MoAb): CD45/CD14, CD3/CD19/CD45, CD4/CD8 / CD3, CD20/CD5/CD3, CD3/CD16-56/CD45, CD2/CD7, FMC7/CD23, CD103/CD22/CD20, HLADR/CD38, CD10/CD19, CD1a, CD11b and also IgM/gD, kappa and lambda immunoglobulin light chains for the detection of surface immunoglobulin and clonal restriction for immunoglobulin light chain. The Hematological data were obtained from the hematological analyzer and cytomorphological analysis in blood film stained by Leishmann. The study samples consisted of 45 men and 35 women, ages ranging from 55 to 84 years (mean 65 years). Complete white blood count showed count ranging from 10.0 to 42.0 x 109/l. (mean 50.0 x 109/l) and lymphocytes count greater than 5.0 x 109/l in all cases. The neoplastic cells displayed B-CLL phenotype (CD5+/CD19+/CD20+/HLADR+/CD23+) in the vast majority of the cases, associated to failed to stain for T cell markers (CD1a, CD2, CD4, CD3, CD7, CD8), CD103, CD14 and FMC7. Leukemic cells of most patients also expressed low intensity of IgM and IgD with restricted kappa light chain, in most cases (59,7%). This observation highlights the importance of immunophenotyping for correct diagnosis of chronic lymphoproliferative syndromes and the panel of MoAb used was sufficient for diagnostic confirmation of B-CLL

Efetividade de métodos de identificação sexual em tilápias-do-nilo (oreochromis niloticus) revertidas sexualmente com hormônio em ração com diferentes granulometrias

Three methods of sexual identification were evaluated in Nile tilapia (Oreochromis niloticus) at ages of 30, 60 and 90 days that were submitted to sexual reversion under administration of androgen hormone 17 alpha-methyltestosterone added to crumble diets with different granules sizes (0.25, 0.35 and 0.50 mm) in dosage of 60mg/kg. The techniques of sexual identification employed were: a) Macroscopic examination of urogenital papilla; b) Microscopic examination of gonads through stain with carmine-acetate and c) Microscopic examination of gonads through histological routine. In the presence of three methods of sexual identification evaluated, histology of gonads has shown the most confident diagnosis. The grain diameters of rations did not interfere in morphological characteristics of gonads, neither in secondary sexual characters of fish.

Astrocytic and microglial response and histopathological changes in the brain of horses with experimental chronic Trypanosoma evansi infection

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

Efeito de Colletotrichum dematium var. truncata e Cercospora kikuchii na germinação de sementes de soja

A antracnose, causada por Colletotrichum dematium var. truncata, é a principal doença da soja que afeta a fase inicial de formação das vagens, podendo causar a morte das plântulas. Cercospora kikuchii é o fungo causador da doença mancha púrpura nas sementes de soja, responsável por severas reduções no rendimento e na qualidade das sementes. O objetivo deste trabalho foi determinar o tempo mais apropriado para infecção das sementes de soja por C. dematium var. truncata e C. kikuchii, para posterior avaliação dos danos causados pelo fungo na germinação. Tais fungos foram cultivados em meio BDA. As sementes de soja cultivar MSOY 6101 foram colocadas sobre meio contendo o patógeno C. dematium var. truncata por 0 (testemunha), 4, 16, 24, 32 e 40h. As sementes da cv. CD 208 foram inoculadas com C. kikuchii pelos mesmos períodos citados, com adição de 48 e 56h. Após os respectivos tempos de contato, as sementes foram submetidas ao teste de sanidade (papel de filtro), com duas variações, sem e com assepsia superficial (hipoclorito de sódio 1% por três minutos). Determinado o tempo mais adequado de infecção, outras sementes foram infectadas pelo patógeno e, posteriormente, foram realizados testes de germinação em papel e areia com uma mistura de sementes sadias (colocadas sobre o meio BDA) e sementes inoculadas, resultando em lotes com 0, 20, 40, 60, 80 e 100% de sementes infectadas. O tempo de incubação de 40 e 56 horas para C. dematium var truncata e C. Kikuchii, respectivamente são suficientes para obtenção da totalidade das sementes infectadas. C. Kikuchii não prejudica a germinação das sementes.

Induction of the acrosome reaction test to in vitro estimate embryo production in Nelore cattle

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

Morphology of glandular stomach (Ventriculus glandularis) and muscular stomach (Ventriculus muscularis) of the partrigde Rhynchotus rufescens

Vinte perdizes Rhynchotus rufescens adultas foram utilizadas para estudo morfológico do proventrículo e ventrículo gástricos da perdiz Rhynchotus rufescens. Os materiais foram coletados e os comprimentos do proventrículo e do ventrículo gástricos foram avaliados. Para o estudo histológico, fragmentos dos estômagos foram corados pelas técnicas de ácido periódico de Schiff (PAS) e tricromo de Masson. O proventrículo gástrico é alongado, com formato fusiforme direcionado no sentido craniocaudalmente e para a esquerda, e apresenta um comprimento médio 3,20cm nas fêmeas e 3,65cm nos machos. Histologicamente, o proventrículo gástrico é composto por vários lobos e glândulas. A mucosa é formada por epitélio cúbico, sendo bastante pregueada. O ventrículo gástrico tem o formato de uma lente biconvexa, com comprimento médio de 4,30cm nas fêmeas e 4,35cm nos machos. A mucosa é formada por pregas revestidas por células cilíndricas e pelo muco formador da cutícula. Há criptas na base das pregas. em seguida, há uma lâmina própria e uma espessa camada muscular lisa, que se encontra direcionada de acordo com o formato do ventrículo gástrico. A serosa é constituída por uma densa porção de tecido conjuntivo, entremeado por algumas células musculares lisas.

Morphology of beak and tongue of partrigde Rhynchotus rufescens

Vinte perdizes Rhynchotus rufescens foram utilizadas para estudar a morfologia do bico e da língua. Os comprimentos do bico e da língua foram avaliados, e secções histológicas foram coradas com Hematoxilina-Eosina (HE), ácido periódico de Schiff (PAS) e tricromo de Masson, posteriormente analisadas e descritas. O bico da perdiz em ambos os sexos são curvos, duros e com uma extremidade pontiaguda tendo em média de 4,90cm de comprimento para as fêmeas e 4,80 para os machos. A língua é caracterizada por um formato triangular e possui em média 1cm de comprimento para ambos os sexos, sendo sua extremidade pontiaguda. A organização histológica da língua apresenta epitélio estratificado queratinizado com papilas filiformes em toda a superfície, glândulas mucosas com ductos que se projetam na superfície epitelial e cartilagem hialina em todo o comprimento, localizada sobre a musculatura esquelética, a qual é orientada em várias direções.

Esporos de Clostridium botulinum em mel comercializado no Estado de São Paulo e em outros Estados brasileiros

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Immunohistochemical characterization of mononuclear cells and MHC II expression in the brain of horses with experimental chronic Trypanosoma evansi infection

Este estudo objetivou caracterizar a resposta imune celular no sistema nervoso central (SNC) de eqüinos com infecção crônica experimental por Trypanosoma evansi. Para este propósito, foram utilizados os métodos histoquímicos (HE) e imunoistoquímicos do complexo avidina-biotina peroxidase (ABC). O fenótipo do infiltrado celular foi caracterizado com o auxílio de anticorpos anti - CD3, para linfócitos T e antiBLA36 para linfócitos B. Os macrófagos foram marcados com anticorpo antiantígenos da linhagem mielóide/histiócitos (Clone Mac387). A lesão no sistema nervoso central (SNC) dos eqüinos infectados com T. evansi foi caracterizada como meningoencefalite e meningomielite não supurativa. A gravidade das lesões variou em diferentes segmentos do SNC, refletindo distribuição irregular das alterações vasculares. A distribuição de células T e B e antígenos do complexo maior de histocompatibilidade classe II foram avaliados dentro do SNC de eqüinos cronicamente infectados com T. evansi. O infiltrado perivascular e meníngeo eram constituídos predominantemente por células T e B. Macrófagos foram raramente visualizados. T.evansi não foi identificado no parênquima do SNC dos eqüinos.

Detection of anti-Toxoplasma gondii antibodies in experimentally and naturally infected non-human primates by Indirect Fluorescence Assay (IFA) and indirect ELISA

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

Disseminated avian tuberculosis in captive Ara macao

Avian mycobacteriosis was diagnosed in a captive scarlet macaw (Ara macao) that presented multifocal granulomas on subcutaneous tissue, sciatic nerves, infraorbital sinus, trachea, air sacs, muscles, spleen and liver. Microscopically, central areas of caseous necrosis surrounded by epithelioid macrophage, multinucleated giant cells, and lymphocytes were observed. Acid-fast bacilli were demonstrated by Ziehl-Neelsen stain. Inoculation into Lowenstein-Jensen, Stonebrink and Petragnani media, yielded Mycobacterium spp, which was identified as Mycobacterium avium by polymerase chain reaction technique (PCR).

Effects of scrotal insulation in Nellore bulls (Bos taurus indicus) on seminal quality and its relationship with in vitro fertilizing ability

The objectives of this study were to assess the effects of induced testicular degeneration in Bos taurus indicus (Nellore) bulls on changes in seminal characteristics and fertilizing ability of sperm. Four Nellore bulls (30-36-month-old, 500-550 kg) with good seminal quality (> 80% motile and morphologically normal sperm) had serotal insulation applied for 5 d. Semen was collected by electroejaculation and cryopreserved at a the pre-insulation moment, and 7, 14, and 21 d after insulation was removed. Gross motility, vigor of sperm movement (1-5), acrosome integrity, sperm morphology (phase-contrast microscopy), nuclear vacuoles and abnormal chromatin (Feulgen-stain) were determined after sperm preparations for in vitro fertilization (IVF). Prior to IVF, sperm were separated using a Percoll gradient (45% and 90%). Normal sperm decreased (P < 0.05) 14 and 21 d after insulation was removed. on 14 and 21 d, the incidence of head defects (9.7 +/- 0.6 and 17.0 +/- 0.8, respectively; mean +/- S.E.M) was higher (P < 0.05) in agreement with the incidence of nuclear vauoles (14.0 +/- 5.0 and 12.3 +/- 2.3) and abnormal chromatin (24.4 +/- 7.2 and 30.8 +/- 2.8). Although the frequency of cleaved oocytes decreased only on 21 d (P < 0.05), blastocyst rates were lower (P < 0.05) than pre-insulation on 14 and 21 d. In regression analyses, only nuclear vacuoles, head defects and intact acrosome accounted for differences in cleavage (R(2) = 0.38, 0.48, and 0.30, respectively) and blastocyst rates (R(2) = 0.35, 0.37, and 0.44). Abnormal chromatin was associated only with blastocyst rates (R(2) = 0.35). In conclusion, blastocyst rate was more sensitive than cleavage rate and the assessment of nuclear integrity is recommended to predict the fertilizing ability of bull sperm. (c) 2008 Elsevier B.V. All rights reserved.

Avaliação, identificação e atividade enzimática de bactérias psicotróficas presentes no leite cru refrigerado

Extended storage of refrigerated milk can lead to reduced quality of raw and processed milk, which is a consequence of the growth and metabolic activities of psychrotrophic bacteria, able to grow under 7oC or lower temperatures. Although most of these microorganisms are destroyed by heat treatment, some have the potential to produce termoresistant proteolytic and lipolytic enzymes that can survive even UHT processing and reduce the processed products quality. Recently, the IN 51 determineds that milk should be refrigerated and stored at the farm what increased the importance of this group of microorganisms. In this work, psychrotrophic bacteria were isolated from 20 communitarian bulk tanks and 23 individual bulk tanks from dairy farms located at Zona da Mata region of Minas Gerais State and from southeastern Rio de Janeiro. Selected milk dilutions were plated on standard agar and after incubation for 10 days at 7oC, five colonies were isolated, firstly using nutrient agar and after using McConkey agar for 24 hours at 21oC. The isolates were identified by morphology, Gram stain method, catalase production, fermentative/oxidative metabolism and by API 20E, API 20NE, API Staph, API Coryne or API 50 CH (BioMerieux). In order to ensure reproductibility, API was repeated for 50% of the isolates. Species identification was considered when APILAB indexes reached 75% or higher. 309 strains were isolated, 250 Gram negative and 59 Gram positive. 250 Gram negative isolates were identified as: Acinetobacter spp. (39), Aeromonas spp. (07), A. Hydrophila (16), A. sobria (1), A. caviae (1), Alcaligenes feacalis (1), Burkholderia cepacia (12), Chryseomonas luteola (3), Enterobacter sp. (1), Ewingella americana(6), Hafnia alvei (7), Klebsiella sp. (1), Klebsiella oxytoca (10), Yersinia spp. (2), Methylobacterium mesophilicum (1), Moraxella spp. (4), Pantoea spp. (16), Pasteurella sp. (1), Pseudomonas spp. (10), P. fluorescens (94), P. putida (3), Serratia spp. (3), Sphigomonas paucomobilis (1). Five isolates kept unidentified. Pseudomonas was the predominant bacteria found (43%) and P. fluorescens the predominant species (37.6%), in accordance with previous reports. Qualitative analysis of proteolytic and lipolytic activity was based on halo formation using caseinate agar and tributirina agar during 72 hours at 21oC and during 10 days at 4°C, 10oC and 7°C. Among 250 Gram negative bacteria found, 104 were identified as Pseudomonas spp. and 60,57% of this group showed proteolytic and lipolytic acitivities over all four studied temperatures. 20% of Acinetobacter, Aeromonas, Alcaligenes, Burkholderia, Chryseomonas, Methylobacterium, Moraxella presented only lipolytic activity. Some isolates presented enzymatic activity in one or more studied temperatures. Among Gram positive bacteria, 30.51% were proteolytic and lipolytic at 10oC, 8.47% were proteolytic at 7oC, 10oC, and 21oC, 8.47% were proteolytic at all studied temperatures (4oC, 7oC, 10oC and 21oC) and 3.38% were proteolytic only at 21oC. At 4oC, only one isolate showed proteolytic activity and six isolates were lipolytic. In relation to Gram negative microorganisms, 4% were proteolytic and lipolytic at 7oC, 10oC and 21oC, 10% were proteolytic at 10oC and 4.4% were lipolytic at 4oC, 7oC, 10oC and 21oC, while 6.4% of all isolates were proteolytic and lipolytic at 10oC and 21oC as well as lipolytic at 4oC and 7oC. These findings are in accordance with previous researches that pointed out Pseudomonas as the predominant psycrotrophic flora in stored refrigerated raw milk

A Nonstaining and Tasteless Hydrophobic Salt of Chlorhexidine

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)