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Objective: The current study aimed to design a histological method to determine the presence and organization of the collagen network in sturgeon notochord. Methods: Serial sections of sturgeon notochord (Acipenser naccarii) were used and assigned to two different experimental groups: Hyaluronidase pre-treatment (HP) in an alcohol acid solution (1% HCl in 70% alcohol solution) for 15 min and hyaluronidase solution in a 2 µg/ ml concentration (pre-heated at 37º C), and control (CTR) group, without pre-treatment. Then, the ECM was assessed by two histochemical methods: Picrosirius Red (PR) staining for 30 min with Sirius red (0.1% of Sirius red in saturated aqueous picric acid), for collagen bundle staining, and Alcian Blue (AB) staining for glycosaminoglycans detection. Results: Samples analyzed in this study showed positive histochemical reaction for collagen fibers in both experimental groups. Referring to PR staining, the CTR group presented a larger and homogeneous reaction was observed in the entire samples, whereas HP group presented a more definite and intense pattern of collagen network. Also, this more intense signal in HP group matched with an increase of birefringence in polarized microscopy images of PR. However, HP group showed a lower intense and more heterogeneous signal when was compared with CTR group in AB staining. Conclusion: Using a simple histological example, our study illustrates the capability of a hyaluronidase pre-treatment to enhance picrosirius red staining in sturgeon notochord trough light and polarized microscopy., Objetivo: El presente estudio tiene por objetivo diseñar un método histológico para determinar la presencia y organización de la red de colágeno en la notocorda del esturión. Métodos: Secciones seriada de la notocorda de esturión (Acipenser naccarii) fueron utilizadas y se asignaron a dos grupos experimentales diferentes: pretratamiento de hialuronidasa (HP) en una solución de alcohol ácido (HCl al 1% en solución de alcohol al 70%) durante 15 minutos y posteriormente a una solución de hialuronidasa en una concentración de 2 μg / ml (precalentada a 37º C) y grupo de control (CTR), sin tratamiento previo. Luego, la ECM se evaluó mediante dos métodos histoquímicos: tinción Picrosirius Red (PR) durante 30 minutos con rojo Sirio (0,1% de rojo Sirio en una solución saturada de ácido pícrico), para la tinción de colágeno; y tinción con Alcian Blue (AB) para detección de glicosaminoglicanos. Resultados: Las muestras analizadas en este estudio mostraron una reacción histoquímica positiva para las fibras de colágeno en ambos grupos experimentales. Con respecto a la tinción PR, el grupo CTR presentó una reacción mayor y más homogénea en toda la superficie de las muestras, mientras que el grupo HP presentó un patrón red de colágeno más definido e intenso. Además, esta señal más intensa en el grupo HP coincidió con un aumento de la birrefringencia en las imágenes de microscopía polarizada de PR. Sin embargo, el grupo HP mostró una señal menos intensa y más heterogénea cuando se comparó con el grupo CTR en la tinción AB. Conclusión: Utilizando un ejemplo histológico simple, nuestro estudio ilustra la capacidad de un pretratamiento de hialuronidasa para mejorar la tinción de picrosirius en la notocorda del esturión a través de la luz y la microscopía polarizada.

The aim of this study was to evaluate the modulation of skeletal muscle fibres from Ile de France lambs fed with levels of whole cottonseed (WCS), as well as to develop sheep meat products and characterise them. This study was composed of three experimental tests. The first evaluated the serum levels, modulation, and performance of skeletal muscle fibres of Longissimus thoracis of Ile de France lambs fed with whole cottonseed and its relation to meat quality. The skeletal muscle fibres were identified by histochemical analysis and classified according to contraction velocity, staining and metabolism. Fifty Ile de France lambs, five references and five treatments (0, 10, 20, 30 and 40% WCS in the diet) were used, with nine replicates and a completely randomised design. The linear and quadratic trends, as well as the Spearman partial correlation between fibre characteristics and meat quality (58 variables) were evaluated, with whole cottonseed levels being used as covariates. Serum cholesterol, total protein and gamma glutamyl transpeptidase levels increased (P0.05) the texture profile, but they negatively affected the acceptability of the hamburgers (P0,05) o perfil de textura, mas afetaram negativamente a aceitabilidade dos hambúrgueres (P

A 4-year-old female captive-bred snake of the genus Bothrops showed swelling on the left side of the oral cavity, suggesting the development of neoplasia. The mass was removed surgically and sent for pathological examination. Two months later a new increase in volume in the same site was observed, suggesting recurrence. The lesion was completely removed and sent for pathological analysis. Histologically, the two-samples consisted of a mass with highly-cell density composed of spindle-shaped anaplastic cells arranged in interwoven bundles, distributed throughout the tissue extension and, occasionally, polygonal cells arranged in irregular fascicles. The Masson trichrome staining showed modest amount of collagen supporting the neoplastic cells. PAS-positive content was not observed in the cytoplasm of neoplastic cells. Histological and histochemical findings indicated that it was a spindle cell neoplasm, but the classification was not possible. Immunohistochemistry was requested and performed using the streptavidin-biotin-peroxidase method. The markers used were anti-vimentin, anti-PCNA, anti-EMA, anti-melan A and anti-melanosome, anti-desmin, anti-actin, anti-CD68 and anti- S100protein. The neoplastic cells were immunoreactive for vimentin and PCNA and negative for the other antibodies. The morphology characterization, histochemical and immunohistochemical analysis of neoplastic cells allowed the definitive diagnosis of oral fibrosarcoma. Resumo Uma serpente de cativeiro, fêmea, quatro anos de idade, do gênero Bothrops apresentou aumento de volume no lado esquerdo da cavidade oral, sugerindo tratar-se de neoplasma. A massa foi removida cirurgicamente e enviada para exame anatomopatológico. Dois meses depois foi observado novo aumento de volume no mesmo local, sugerindo recidiva. A lesão foi removida por completo e também enviada para análise. Histologicamente, as duas amostras consistiam de massa altamente celular, composta por células anaplásicas fusiformes organizadas em feixes entrelaçados e distribuídos por toda extensão tecidual e, ocasionalmente, células poligonais arranjadas em fascículos irregulares. A coloração de tricrômico de Masson apresentou quantidade modesta de colágeno sustentando as células neoplásicas. Não foi observado conteúdo PAS-positivo no citoplasma das células neoplásicas. Os achados histológicos e histoquímicos indicavam tratar-se de neoplasma de células fusiformes, porém não era possível sua classificação. A imuno-histoquímica foi requisitada e realizada pelo método streptavidina-biotina-peroxidase, utilizando os anticorpos anti-vimentina, anti-PCNA, Anti-EMA, anti-melan A, anti-HMB45, anti-desmina, anti-actina, anti-CD68 e anti-proteína S-100. As células neoplásicas foram imunorreativas para vimentina e PCNA e, negativas para os demais anticorpos. A caracterização morfológica, histoquímica e imuno-histoquímica das células neoplásicas permitiu o diagnóstico definitivo de fibrossarcoma oral.

Study on resistance mechanism to vascular-streak dieback (VSD) disease in cacao (Theobroma cacao L.) is limited due to the lack of fungal spores for artificial inoculation. This research was conducted to study the production of secondary metabolites that appear to be evidence of defense signaling in resistant clone of Sca 6 and susceptible clone of TSH 858 to Ceratobasidium theobromae natural infection. A fungal staining method was employed to detect C. theobromae hyphae at early infection stages, before VSD symptoms appear. Metabolite profiling was analyzed using pyrolysis gas chromatography mass spectrometry (Py-GCMS) at pre-, early and late stages of C. theobromae infection. Histochemical and anatomical characteristics of both healthy and infected leaves were also observed to identify the accumulation sites of secondary metabolites on and in cocoa leaf tissues. The results confirmed that fungal staining using trypan blue can detect early stages of C. theobromae infection; at the 14th week (on susceptible seedlings) and the 18th week (on resistant clones), following placement of the seedlings under infected cacao plants. Phenylpropanoid biosynthesis, terpenoid biosynthesis, environmental information processing signal transduction pathways, and aromatic biodegradation were detected as important metabolite pathways during defense mechanism. I-limonene (terpenoid), p-ethylguaiacol (phenols) and 2.3 dihidrobenzofuran (heterocyclic compounds) were proposed as an active defense produced by the host after infected by pathogen mainly on late infection of C. theobromae. Terpenoid and phenol compounds were accumulated on the glandular trichomes, idioblast of upper and bottom epidermis, phloem vessel and cortex idioblast of cacao leaves. Epidermis thickness of resistant clone was significantly greater than that of susceptible clone on both surfaces. Leaf epidermis tissue and the accumulated compounds in epidermis idioblast may act as the physical and biochemical markers of cocoa resistance to VSD.

Amphilophium crucigerum (L.) L.G. Lohmann, known as “pente-de-macaco” is a species of Bignoniaceae native to Brazil, and whose seeds are used in folk medicine. This study aimed to describe morphoanatomical features of this species of fruit to aid in its correct identification and pharmacognostic analysis. Samples of ovary, pericarp and seed were fixed with 3% glutaraldehyde, sectioned on a rotary microtome and analyzed by stereomicroscope. The results are shown in three parts: (1) The ovary presents peltate trichomes, long non-glandular trichomes and emergences in the epidermis; it is 2-carpellate and unilocular with two intruding parietal placenta; ovules are numerous on the placenta; it presents a large quantity of crystals. (2) The pericarp is woody, densely echinate and elliptic shape; it presents a 2-valved capsule and is septicidal; it presents emergences, stomata, lenticels, crystals and a large quantity of clustered stones cells. (3) Seeds are alate, exalbuminate and exotestal; there is a large amount of crystals in the exotestal region; it presents an endothelium and remnant endosperm. Histochemical tests showed the presence of lipophilic substances, polysaccharides, phenolic substances, alkaloids and a small quantity of starch. These pharmacobotanical features described for A. crucigerum are essential for the pharmacognostic analysis of the drug plant. Keywords: Crystals, Histochemical, Morphoanatomy, Ovary, Seed, Trichomes

This study investigated if overfed rats present morphological and histochemical muscle adaptation similar to normally fed, both submitted to two different weekly frequencies of training. Thirty male Wistar rats were fed either with standard chow (SCØ) or with hypercaloric diet (HCØ). They were subdivided into six subgroups: sedentary (SCØ and HCØ), trained twice/week (SC2 and HC2) and trained five times/week (SC5 and HC5). The trained groups swam 60 min/day, during 10 weeks. Twenty four hours after the last training, samples of Gastrocnemius were excised and stained with HE, NADH-TR and m-ATPase, and the capillary density was calculated. Total heart mass (HM) and the mass of atrium (AM), left (LV) and right (RV) ventricles were excised and weighted. The comparisons were made by ANOVA and by Covariance analysis, adjusting the variables by body weight. The results showed that the HCØ achieved higher BM, however, absolute HM did not differ post training. Irrespective of the diet, rats that were trained twice a week presented significantly greater increase in the AM. In general, the SC5 and HC5 groups showed higher HM, LV, RV, proportion of oxidative fibres and capillary density, compared to the sedentary and twice week trained groups. A higher proportion of injuries (splitting) was noted in the HC2 and HC5 compared to SC2 and SC5. These results indicate that the frequency of training influenced the skeletal and heart adaptation and larger changes were observed in the 5x/week group, which ingested the standard diet. The 5x/week training groups also presented large amount of muscle fibres damage.

In this study, possibility of using Skipjack Tuna (Katsuwonus pelamis) fish skins as a new and alternative raw material source for leather industry was investigated. The structural and chemical properties of fish skins have a high impact on the preparation of special production recipes and the manufacturing of high performance valuable fish leathers. For this purpose, characteristic properties of Skipjack Tuna fish skins were evaluated by histological, histochemical and chemical methods. The chemical characteristics of fish skins were determined by analyzing fat content (%), total Kjeldahl nitrogen (%), hide substance (%), volatile matter (%), total ash (%) and fatty acid methyl esters and total amino acid content through instrumental analysis. Histological and histochemical analyses were carried out by Hematoxylin-Eosin and Van Gieson staining techniques. As a result of histological and histochemical investigations, fish skins were found thin and tough due to their surface characteristics, had less collagen content compared to other raw materials commonly used in leather industry and the fibers were orientated in parallel but opposite to striated muscles differently from the other skins. Besides, the fat content, Kjeldahl nitrogen, hide substance, volatile matter and total ash of Katsuwonus pelamis fish skins were found 24.47±1.76%, 10.20±0.28%, 57.32±1.55%, 37.33±4.01% and 1.23±0.0002% respectively. The results of the study revealed that Katsuwonus pelamis fish skins could be an alternative raw material to be used in the production of high value added luxury leathers.

Made available in DSpace on 2018-12-11T17:29:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Sugarcane vinasse is one of the main residues generated by the transformation of cane into ethanol. Because of the high organic content (COD), high biochemical oxygen demand (BOD), low pH, the large amount that this residue is generated (15 l for every liter of ethanol produced) and their use as fertilizer on the sugarcane crop, this residue is potentially polluting to the soil ecossystem and by percolation to water ecossystem too. Thus, this study aimed to assess the toxicity of vinasse by analyzing Oreochromis niloticus gills exposed to different dilutions (1%, 2.5%, 5% and 10%) in two bioassays. The gills were collected, fixed and analyzed using ultra morphological, histological, and histochemical techniques. After exposure to the vinasse, a statistically significant reduction of the ridges present on the surface of pavimentous cells was observed in one of the bioassays; such structures are responsible for mucus retention, which helps to protect the tissue. In addition, an intumescence of the cells was observed in the treatments with vinasse as well as an increase in the amount of chloridric cells. Some striking tissue changes detected in the treatments were epithelial detachment and loss of integrity of secondary lamellae, causing their rupture and consequent hemorrhage. In the first bioassay, the amount of these changes was statistically significant at the 5% dilution, and the focus of hemorrhage was significant at all dilution ratios. In the second bioassay, the epithelial disorganization was statistically significant only at the 2.5% dilution of vinasse. Moreover, for both bioassays performed, a significant increase in mucous cells was observed when compared with the control. Our results demonstrate the toxic action of sugarcane vinasse, which caused histopathological changes in the exposed animals at all four dilution tested. This highlights the need for caution in the disposal of sugarcane vinasse on the soil, especially due to its capacity for being leached or percolated into water resources, which could seriously damage aquatic fauna. UNESP (Universidade Estadual Paulista), Av. 24-A, 1515. Bela Vista Centro Universitário Hermínio Ometto FHO-UNIARARAS Av. Dr. João Maximiliano Baruto 500. Jd. Universitário. UNESP (Universidade Estadual Paulista), Av. 24-A, 1515. Bela Vista FAPESP: 2011/0679-8 FAPESP: 2012/50197-2

There are report two human envenenomations for species from Clelia Fitzinger, 1826 (opisthoglyph snake). The patients exhibited symptoms similar from bothropic accident. Then this work have with objective, the hystological and histochemical study of Duvernoy's gland from Clelia plumbea. For the hystological observation of the Duvernoy's gland were using, with paraffin's cutting edge, two methods: Hematoxylin + Eosin and Toluidine's Blue. With historesine's cutting edge were using three methods: Metilen's Blue, Toluidine's Blue + Floxine and Toluidine's Blue. For the histochemical detection were using six techniques, with paraffin's cutting edge: PAS, PAS + Alcian Blue pH 2,5, Alcian Blue pH 2,5, PAS + Salivar Amilase, Method of Lars Grimelius and Method of Lillie. It was evidenced the predominance of serous cells on Duvernoy's gland. In relation to the histochemical constituion of the duvernoy's gland, was verified the presence of glucoconjugates neuters, enzymes and cells with reduetive activity. By fact from this species to kill his preys by constriction, the predominance of serous cells in the duvernoy's gland, don' t must be direct related with the subjugation of his preys, but with the digestive processes. Histological and histochemical analysis showed cells of Duvernoy's gland are constituted for two populations: serous cells (prodution of enzymes) and mucous cells (prodution of glucoconjugates).

There are report two human envenenomations for species from Clelia Fitzinger, 1826 (opisthoglyph snake). The patients exhibited symptoms similar from bothropic accident. Then this work have with objective, the hystological and histochemical study of Duvernoy's gland from Clelia plumbea. For the hystological observation of the Duvernoy's gland were using, with paraffin's cutting edge, two methods: Hematoxylin + Eosin and Toluidine's Blue. With historesine's cutting edge were using three methods: Metilen's Blue, Toluidine's Blue + Floxine and Toluidine's Blue. For the histochemical detection were using six techniques, with paraffin's cutting edge: PAS, PAS + Alcian Blue pH 2,5, Alcian Blue pH 2,5, PAS + Salivar Amilase, Method of Lars Grimelius and Method of Lillie. It was evidenced the predominance of serous cells on Duvernoy's gland. In relation to the histochemical constituion of the duvernoy's gland, was verified the presence of glucoconjugates neuters, enzymes and cells with reduetive activity. By fact from this species to kill his preys by constriction, the predominance of serous cells in the duvernoy's gland, don' t must be direct related with the subjugation of his preys, but with the digestive processes. Histological and histochemical analysis showed cells of Duvernoy's gland are constituted for two populations: serous cells (prodution of enzymes) and mucous cells (prodution of glucoconjugates).

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) A miopatia centronuclear (MCN) é uma forma rara de miopatia congênita. De acordo com a época do início dos sinais e sintomas e com o grau de envolvimento muscular são distinguidas três formas clínicas: forma neonatal severa; forma de início na infância; e de início na vida adulta. São apresentados neste estudo os achados histopatológicos de dez pacientes portadores da forma de início na infância da MCN. Os fragmentos musculares foram processados através de colorações de rotina e histoquímica, e em três casos foi realizado estudo ultraestrutural. Dentre os resultados obtidos, destacou-se o aumento da centralização nuclear na fibra muscular, que variou de 25 a 90%. Adicionalmente, foram observadas predominância de fibras do tipo I, variabilidade entre o diâmetro das fibras musculares, alterações da arquitetura interna das fibras musculares e presença de áreas focais de desorganização dos miofilamentos. Devido a estes aspectos, os principais diagnósticos diferenciais considerados foram as miopatias por predominância de fibras e por desproporção de fibras, e a distrofia miotônica. As anormalidades histológicas observadas na MCN podem ser devidas a uma parada no processo maturacional do músculo esquelético na fase miotubular fetal. A causa deste defeito ainda permanece sem explicação completa. Centronuclear myopathy is a rare congenital myopathy. According to the period of onset of signs and symptoms and the degree of muscular involvement three clinical forms are distinguished: severe neonatal; childhood onset; and adult onset. We describe herein the muscle biopsy findings of ten patients with the childhood onset form of the disease including three cases with ultrastructural study. The biopsies disclosed increased nuclear centralization that varied from 25 to 90% of the fibers, type 1 predominance, great variability in fiber diameters, involvement in the internal fiber's architecture, and focal areas of myofilament disorganization. The main histopathologic differential diagnoses included type I fiber predominance, congenital fiber type disproportion, and myotonic dystrophy. The histologic abnormalities in centronuclear myopathy may be due to an arrest of maturation on the fetal myotubular stage. The cause of this arrest remains elusive. Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Department of Neurology and Neurosurgery UNIFESP-EPM Department of Pathology UNIFESP, EPM, Department of Neurology and Neurosurgery UNIFESP, EPM Department of Pathology SciELO

Descreveram-se os sinais clínicos e achados anatomopatológicos da intoxicação crônica por cobre em um ovino da raça Texxel e definiu-se a conduta diagnóstica correta para confirmação da enfermidade. Um ovino foi encaminhado ao setor de patologia com histórico de apatia, hemoglobinúria e morte em dois a três dias. No exame necroscópico, observaram-se icterícia e edema subcutâneo, fígado aumentado de volume e amarelado e rins escuros. No exame histológico, observaram-se necrose zonal aleatória e acentuada no fígado, necrose epitelial tubular, gotas hialinas e cilindros marrom-alaranjados em túbulos coletores dos rins. O histórico alimentar, a sensibilidade de espécie/raça, o quadro clínico, as alterações macroscópicas e microscópicas sugeriram o quadro de intoxicação crônica por cobre. A confirmação diagnóstica somente foi possível após a marcação de pigmentos de cobre pela técnica histoquímica de Ulzmann e pela quantificação de cobre em matéria seca de fígado e rins, cujos valores foram mais altos que o normal. The present work describes the clinical signs and anatomopathological findings of chronic copper toxicities in a Texxel breed sheep and defines the optimal diagnostic procedure for confirmation of the disorder. A sheep was sent to pathology analysis service with a history of apathy, hemoglobinuria and death within two to three days. Necropsy showed jaundice and subcutaneous edema, enlarged yellow liver and dark kidneys. The histologic examination showed random zonal necrosis, marked necrosis in the liver and tubular epithelial and orange-brown spotted hyaline cylinders in the collecting tubules of the kidneys. The dietary history, sensitivity of species/breed, clinical, macroscopic and microscopic alterations suggested the framework of chronic copper poisoning. Diagnostic confirmation was only possible after staining copper pigments trough the Ulzmann technique and quantification of copper in the dry liver and kidney, which were higher than normal levels.

Submitted by isabela.moljf@hotmail.com (isabela.moljf@hotmail.com) on 2017-08-31T15:40:53Z No. of bitstreams: 0 Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-09-01T11:35:40Z (GMT) No. of bitstreams: 0 Made available in DSpace on 2017-09-01T11:35:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2013 CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Por alinhamento das sequências de aminoácidos da apirase de S. tuberosum e isoformas de outras espécies de plantas, alta identidade (46% a 94%) e similaridade (64% a 96%) foram identificadas entre elas, incluindo espécies diferentes de Fabaceae, e estreita relação estrutural foi encontrada entre os domínios B destas proteínas. Anticorpos policlonais anti-apirase de batata S. tuberosum inibiram 65-80% da atividade catalítica desta proteína. Soros imunes policlonais produzidos contra os peptídeos sintéticos homólogos potB1LJ, LbB1LJ e SmB1LJ ou potB2LJ, LbB2LJ e SmB2LJ, derivados de domínios B de isoformas de NTPDases, inibiram significativamente (32-44%) a atividade da apirase de batata. Estes anticorpos podem ser ferramentas moleculares auxiliares para o estudo das isoformas de apirase. A atividade apirásica de tecidos de folha (NL) ou caule (NS) normais e de galha globosa (GG) de Calliandra brevipes BENTH (Fabaceae: Mimosoidae) foi testada em meio de reação pH 7,5 e na presença de C12E9, e a hidrólise de ADP, UDP ou GDP por GG foi 1,8 a 2,4 vezes maiores que tecidos não galhados. Anticorpos policlonais anti-apirase de batata inibiram significativamente (70-90%) a hidrólise de ATP, ADP, GDP ou UDP por NS. Em GG, similarmente, a atividade ATPásica, GDPásica, ADPásica ou UDPásica foi significativamente inibida (40-66%). Para o isolamento e identificação de apirase de tecidos normais e galha globosa de Calliandra brevipes, amostra de tecido de NS ou NL não galhado, ou de GG previamente homogeneizada em Triton X-100 mais deoxicolato de sódio foi fracionada em gel não desnaturante contendo os mesmos detergentes. Usando ADP como o substrato, a atividade catalítica da enzima permitiu o aparecimento de uma única banda de precipitado de fosfato de cálcio mostrando idêntica mobilidade eletroforética nas amostras NS, NL e GG, sendo mais intensa nesta última, sugerindo que a apirase é superexpressa neste tecido. Nesta banda de maior mobilidade, distintos polipeptídeos de 52 a 75 kDa foram reconhecidos pela imunoreatividade cruzada de anticorpos anti-apirase de batata, sugerindo a existência de formas ativas da mesma proteína com diferentes cargas líquidas, possivelmente como um resultado de diferentes níveis de glicosilação. Além disso, uma banda de aproximadamente 52 kDa, de menor mobilidade e atividade, foi também identificada em todos os tecidos, possivelmente a apirase em sua forma completamente deglicosilada. Usando técnicas histoquímicas e microscopia óptica, cortes micrométricos corados por lugol ou azul de astra-safranina evidenciaram a câmara pulpal amplamente circundada por tecido nutritivo contendo amido em sua porção distal, e parênquima cortical delimitado por anéis esclerenquimáticos lignificados. Por técnicas histoquímicas, a atividade ADPásica ou ATPásica foi encontrada como um depósito eletrodenso granular de fosfato de chumbo distribuído na superfície externa, e nos tecidos nutritivos da galha globosa. Por técnicas imunohistoquímicas, usando Microscópio Confocal de Varredura a Laser e anticorpos anti-apirase de batata e anti-peptídeos sintéticos, a apirase de galha globosa foi encontrada nos mesmos sítios. Hipóteses funcionais para esta proteína estão sob investigação. Além disso, o tempo de germinação de semente de C. brevipes foi determinado, e no quinto dia após a instalação do teste, a primeira folha foi aberta, identificando tecidos de crescimento rápido apropriados para a investigação da regulação da hidrólise de ATP por apirase na proliferação celular. By alignment of amino acid sequences of the S. tuberosum apyrase and isoforms from other plant species, high identity (46 to 94%) and similarity (64 to 96%) were identified between them, including distinct species of Fabaceae, and closer structural relationship was found between the domains B from these proteins. Polyclonal antibodies anti- S. tuberosum potato apyrase inhibited 65-80% of the catalytic activity of this protein. In addition, polyclonal immune sera produced against homologue synthetic peptides potB1LJ, LbB1LJ and SmB1LJ or potB2LJ, LbB2LJ and SmB2LJ, derivatives from B domains of NTPDases isoforms, significantly inhibited (32 to 44%) potato apyrase activity. These antibodies could be useful molecular tools for studies of the apyrase isoforms. The apyrase activity from the non-galled stem (NS), non-galled leaf (NL), or globose gall (GG) tissue from Calliandra brevipes BENTH (Fabaceae: Mimosoidae) was tested in reaction medium pH 7.4 and in the presence of C12E9, the ADP, UDP or GDP hydrolysis by GG was 1.8 to 2.4 fold higher than non-galled tissues. Polyclonal antibodies anti-potato apyrase significantly inhibited (70 to 90%) the ATP, ADP, GDP or UDP hydrolysis by NS. In GG, similarly, the ATPase, GDPase, ADPase or UDPase activity was also significantly inhibited (40-66%). For isolation and identification of apyrase from normal tissues and globose gall from the Calliandra brevipes, sample of NS, NL or GG tissue was previously homogenised in Triton X-100 plus sodium deoxycholate, and fractionated in non-denaturing gel containing the same detergents. Using ADP as the substrate, the enzyme catalytic activity gave rise to the appearance of a single calcium phosphate precipitate band displaying identical electrophoretic mobilities in NS, NL, and GG samples, being more intense in this last one suggesting that apyrase is over-expressed in this tissue. In this band of higher mobility, a number of distinct polypeptides ranging from 52 to 75 kDa were recognized by cross-immunoreactivity with polyclonal anti-potato apyrase antibodies, suggesting the existence of active forms of the same protein with different net charges, possibly as a result of distinct glycosylation levels. In addition, an immunoreactive band of approximately 52 kDa, of lower mobility and activity, was also identified in all tissues, possibly apyrase in completely deglycosylated form. By histochemical techniques and light microscopy, micrometric sections stained with lugol or Astra Blue-Saphranine evidenced the chamber pulpal widely surrounded by nutritive tissue containing starch in its distal portion, and cortical parenchime delimited by rings sclereides lignified. By histochemical techniques, the ADPase or ATPase activity was found as a granular dense lead phosphate deposit distributed at the external surface, and inside of the nutritive cells of the globose gall. By immunohistochemical techniques, using confocal laser scanning microscopy and antibodies anti-potato apyrase and anti-synthetic peptides, the gall globose apyrase was found at same sites. In addition, the time of laboratory germination of C. brevipes seed was determined, and on the fifth day after test installation, the first leaf opened, identifying rapid growing tissues suitable for further investigations on the participation of both apyrase and regulation of ATP hydrolysis in cell proliferation.

Fundação de Amparo a Pesquisa do Estado de Minas Gerais In ants there are several exocrine glands that vary in their structure, chemical and functional aspects, and the abundance of exocrine glands may be animportant point of their biological success. This work investigated intramandibular glands of "primitive" ants included in the Tribe Ponerini (Ponerinae) and derived Attini (Myrmicinae) including their development in immature, diferentiation between caste and identification of chemical components, contributing to understand the mechanisms involved in the communication of ants as well as to elucidate the function of the intramandibular glands thet is still unknown. The study of the post-embryonic development of the intramandibular glands was performed in Pachycondyla verenae, while the study on the leaf-cutter ant Atta laevigata investigated the occurrence of intramandibular glands in the castes of queen, worker and soldier. The study of the chemical components present in the mandible and body occurred in Pachycondyla villosa. Ants in the study, epidermal secretory cells were found in class I, unicellular glands in class III and epithelial cells with secretory reservoir. The epidermal cells secretory of class I and unicellular glands of class III were found in P.verenae. These glands differentiate during pupation, and their development begins in pink-eyed pupa and is complete in black-eyed pupae. Unlike the gland class that was found in Pachycondyla, the intramandibular glands in A. laevigata, did have in epidermal secretory cells of class I, but had unicellular glands of class III and epidermal cells secretory with reservoir. The morphological, histochemical and morphometric study of the intramandibular glands in A. laevigata supported the hypothesis that these glands are involved directly in the division of labor between castes, and consequently in their chemical communication. The behavioral analyses indicate that chemicals substances from the intramandibular glands of P. villosa are involved in the recognition of nestmates, and the presence of hydrocarbons and cholesterol and the sitosterol found in the mandible of queen may be associated with the profile of caste, likely to be the sitosterol plays role as a flag of fertility and/or a possible source of the hormone. The knowledge of ontogeny, morphology, physiology and chemical composition of the intramandibular glands of ants can contribute to the elucidation of the mechanisms that involve the exocrine system and its relationship with the behavioral aspects and phylogenetic in different subfamilies of Formicidae. It is possible to infer that the intramandibular glands of different tribes and caste produce substances of different natures, indicating different functions, depending on the chemical nature of cellular components. As formigas são especialmente ricas em glândulas exócrinas que variam na sua natureza estrutural, química e complexidade funcional, e esta riqueza em glândulas exócrinas talvez seja o grande ponto de seu sucesso biológico. Este estudo focalizou as glândulas intramandibulares de formigas primitivas Ponerini (Ponerinae) e derivadas Attini (Myrmicinae) quanto ao desenvolvimento de formas imaturas, diferenciação entre castas e componentes químicos, contribuindo para a compreensão dos mecanismos envolvidos na comunicação das formigas na tentativa de entender a possível função da glândula intramandibular que ainda permanece desconhecida. O estudo do desenvolvimento pós-embrionário das glândulas intramandibulares foi realizado em Pachycondyla verenae, enquanto o estudo com a formiga cortadeira Atta laevigata investigou a ocorrência de glândulas intramandibulares nas castas de rainhas, operárias e soldados. Também foi realizado, o estudo dos componentes químicos presentes no interior da mandíbula e no corpo de operárias e rainhas de Pachycondyla villosa. Foram encontradas células epidérmicas secretoras da classe I, glândulas unicelulares da classe III e células epiteliais secretoras com reservatório. As células epidérmicas secretoras da classe I e glândulas unicelulares da classe III, encontradas em P. verenae, diferenciam-se durante a pupação, iniciando seu desenvolvimento na pupa de olho rosa e estão completamente formadas a partir da pupa de olho preto. Diferente da classe glandular que foi encontrada em espécies de Pachycondyla, as glândulas intramandibulares em A. laevigata não apresentaram células epidérmicas secretoras da classe I, mas apresentaram glândulas unicelulares da classe III e células epidérmicas secretoras com reservatório. Os estudos morfológicos, histoquímicos e morfométricos das glândulas intramandibulares em A. laevigata suportam a hipótese de que essas glândulas estejam envolvidas diretamente na divisão de trabalho entre as castas e, consequentemente, na comunicação química. As evidências comportamentais indicam que as substâncias químicas das glândulas intramandibulares de P. villosa estão envolvidas no reconhecimento de companheiras de ninho, e a presença de hidrocarbonetos e os esteróis colesterol em operárias e o sitosterol encontrado na mandíbula de rainhas podem estar associados ao perfil da casta, sendo provável que o sitosterol assuma o papel de um sinalizador de fertilidade e/ou uma possível fonte de hormônio. O conhecimento da ontogenia, morfo-fisiologia e composição química das glândulas intramandibulares das formigas pode contribuir para a elucidação dos mecanismos que envolvem o sistema exócrino e suas relações com os aspectos comportamentais e filogenéticos nas diferentes subfamílias de Formicidae. Com isso, é possível inferir que as glândulas intramandibulares das diferentes tribos e castas produzem substâncias de naturezas distintas, indicando funções diversas, dependendo da natureza química de constituintes celulares.

Conselho Nacional de Desenvolvimento Científico e Tecnológico The aims of this work were to study the germination process of macaw palm seeds through the activities of the enzymes β-mannanase and α-galactosidase, and quantify the amount of mannan present in the endosperm cells. In addition, we studied the dynamics of mobilization of lipid, protein and starch reserves by biochemical and histochemical analysis. The seeds were submitted to a protocol to overcame the species dormancy (patent application: PI0703180-7) and macaw palm germination process was followed up to 29 days. Analyses were performed in embryo / haustorium and lateral and micropylar endosperm separately. The activities of the enzymes β-mannanase and α-galactosidase were performed by colorimetric methods in a spectrophotometer using guar gum and pnitrophenyl-D-galactopyranoside (pNPGal), as substrates, respectively. The content of mannan was analysed by gas chromatography, the lipid by the gravimetric method in the extraction apparatus, the starch by the phenol-sulfuric and soluble protein by Breadford method. Histochemical tests were performed to corifosfina of pectin, cellulose material to calcofluor, Sudan Black B for total lipids, lugol for detection of starch, XP for protein and safrablau for staining and characterization. The activities of β-mannanase and α-galactosidase were higher in the endosperm to the embryo / haustorium throughout the germination period studied. The activities of both enzymes increased in the endosperm after the protrusion of the cotyledon petiole but in the embryo/ haustorium only the β-mannanase activity increased. At the same time, the content of mannan reduced in the endosperm. The lipid content reduced in the embryo from the protrusion of the cotyledon petiole, but increased in the endosperm during the period studied. The Sudan Black B test showed a reduction in number and size of lipid bodies in the embryo. Right after the protrusion of the cotyledonary petiole were identified starch grains, marked with Lugol test, being formed in the haustorium and increasing in quantity until the end of evaluations. In this phase the starch content in the embryo / haustorium increased exponentially. The starch in the endosperm was not identified by lugol histochimical test. The protein content decreased continuously in the embryo / haustorium from the start of imbibition, however, has not changed in the endosperm. The XP test showed qualitative loss of staining of protein bodies in the embryo / haustorium over time. The results suggest that the two enzymes studied are mainly involved in the mobilization of mannans and / or galactomannans of the endosperm after germination. The embryo possesses reserves that provide energy to the early metabolic events of germination. In the endosperm, lipid and protein reserves have not been accessed during the period studied, but there was intense degradation of cell wall polysaccharides from the endosperm side of germination in the strict sense. Os objetivos desse trabalho foram estudar o processo germinativo de macaúba por meio das atividades das enzimas degradativas de mananos, e quantificar o teor de mananos presentes nas células do endosperma. Além disso, estudou-se a dinâmica de mobilização das reservas lipídicas, protéicas e amiláceas por meio de análises bioquímicas e histoquímicas. As sementes foram submetidas ao protocolo de superação de dormência para espécies do gênero Acrocomia (registro de patente: PI0703180-7) e o processo germinativo de macaúba foi acompanhado durante 29 dias. As análises foram realizadas em embrião ou haustório e endosperma adjacente separadamente. As atividades das enzimas de degradação de mananos foram realizadas por métodos colorimétricos em espectrofotômetro utilizando-se como substratos goma-guar, para o ensaio de β-mananase e β-manosidase, e para-nitrofenil α-D-galactopiranosídeo (pNPGal), para o ensaio de α- galactosidase. O teor de mananos foi realizado por cromatografia gasosa, o delipídios por método gravimétrico em aparelho de Soxhlet, o de amido por método fenol-sulfúrico e o de proteínas solúveis por reagente de Bradford. Foram realizados os testes histoquímicos de corifosfina para pectinas, Sudan Black B para lipídios totais, lugol para detecção de amido, XP para proteínas e safrablau para coloração e caracterização. As atividades de β- mananase, β-manosidase e α-galactosidase foram maiores no endosperma que no embrião ou haustório durante todo o período germinativo estudado. As atividades das enzimas aumentaram no endospermaapós a protrusão do pecíolo cotiledonar enquanto que no embrião/haustório somente a atividade da β-mananase e β-manosidase foi aumentada. Ao mesmo tempo, o teor de mananos no endosperma foi reduzido. O teor de lipídios foi reduzido no embrião a partir da protrusão do pecíolo cotiledonar, mas aumentou no endosperma durante o período estudado. O teste de Sudan Black B demonstrou a redução de quantidade e tamanho dos corpos lipídicos no embrião. A partir da protrusão do pecíolo cotiledonar foram identificados grãos de amido, marcados com o teste de lugol, sendo formados no haustório e aumentando em quantidade até o fim das avaliações. O teor de amido no embrião/haustório aumentou exponencialmente a partir da protrusão do pecíolo cotiledonar. Não foi detectado amido pelo teste de lugol no endosperma. O teor de proteínas decaiu continuamente no embrião/haustório desde o início da embebição, entretanto, não foi alterado no endosperma. O teste de XP demonstrou perda de coloração dos corpos protéicos no embrião/haustório ao longo do tempo. Os resultados sugerem que as enzimas estudadas estão principalmente envolvidas na mobilização de mananos e/ou galactomananos do endosperma após a germinação. O embrião possuiu reservas que provêem energia para os primeiros eventos metabólicos da germinação. No endosperma, as reservas lipídicas e protéicas não foram acessadas durante o período estudado, mas houve intensa degradação de polissacarídeos de parede celular do endosperma lateral a partir da germinação no sentido estrito. As modificações bioquímicas e anatômicas citadas resultaram na formação de amido transiente no haustório.

Para la realización del presente estudio, se tomaron biopsias musculares por punción percutánea al M. G. medius, así como muestras de sangre a cincuenta y un CPSC (3-4 años de edad). Dieciséis de estos animales (Grupo I), se encontraban clínicamente sanos e inactivos, y los treinta y cinco restantes (Grupo II), se encontraban en diferentes fases de la rabdomióliss por ejercicio (15 en la fase sub aguda, 3 en la fase aguda y 17 en la fase crónica). Los resultados histoquímicos de las muestras de músculo esquelético del Grupo II, reveló en comparación al Grupo I, un daño preferencial en las fibras de contracción rápida Tipo II, disminución en la capilaridad, reflejado por la disminución significativa (P

Los Caballos Pura Sangre usualmente son entrenados a partir de una edad temprana y en muchos estudios se ha dificultado separar los cambios en el músculo debido al entrenamiento de aquellos producidos por el crecimiento. Se utilizó un grupo de 46 caballos pura sangre venezolanos (41 yeguas y 5 machos) que se encontraban en condiciones de pre-entranmiento o inactivos al menos por un período de 3 meses (mantenidos en el haras con fines reproductivos). La técnica de biopsia por aguja se usó para obtener muestras musculares del M.G. medius. Los caballos se agruparon por edades: Grupo I: 1-2 años n = 5); Grupo II: 3-4 años (n = 10); Grupo III: 5-10 años (n= 19) y Grupo IV: 11-21 años (n =12). Reacciones para la ATPasa a pHs 10,3; 4.6 y 4,37 se usaron para tipificar las fibras; NADH-diaforasa (NADH-d) se utilizó para analizar la capacidad oxidativa; α-glycerophosphato deshydrogenasa (α-GPDH) para estudiar la capacidad glicolítica y la reacción histoquímica cara la α-amilasa PAS para examinar los capilares y medir el grosor de las fibras. Se realizaron determinaciones bioquímicas de las enzimas β-hydroxiacil-CoA-dehydrogenasa (HAD), hexoquinase (HK) y citrato sintasa (CS). Los caballos del Grupo II presentaron 18.6 ± 6,0% fibras tipo 1, 48.1 ± 6,1 % tipo IIA 40,0 ± 11,9 tipo IIB. La proporción de las fibras no difirió de los otros grupos, pero el cociente IIA/IIB fue más alto en el Grupo II que en el Grupo I. El grosor de las fibras no cambió significativamente con la edad. El porcentaje de fibras con reacción alta para la NADH-d fue mas alto en el Grupo II (P

Submitted by (edna.saturno@ufrpe.br) on 2016-11-28T13:38:03Z No. of bitstreams: 1 Maria Esmeralda Cavalcante de Sousa.pdf: 1237941 bytes, checksum: 5d11b8c813aa997680f772fa30705f65 (MD5) Made available in DSpace on 2016-11-28T13:38:03Z (GMT). No. of bitstreams: 1 Maria Esmeralda Cavalcante de Sousa.pdf: 1237941 bytes, checksum: 5d11b8c813aa997680f772fa30705f65 (MD5) Previous issue date: 2009-02-02 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES The cotton leafworm, Alabama argillacea (Hübner) (Lepidoptera: Noctuidae) is an important cotton pest in Brazil, occurring almost in the whole growing season and causing economic losses between 21 and 35%. Bt cotton expressing Cry1Ac protein, a toxin from the bacterium Bacillus thuringiensis (Berliner) (Bt), has recently been released in Brazil. It is resistant to the main cotton lepidopteran pests occurring in Brazil. Bt toxins must interact with a receptor to trigger a toxic effect, although other endogenous factors may account for the differential Bt toxins selectivity towards insects. Therefore, studies on the Cry1Ac interaction with the cotton pests midguts are important to understand insect responses when exposed to the Bt cotton. Then, the aim of this work was to provide an histochemical, morphological, and ultrastructural description of the midgut of A. argillacea larvae fed with the isoline cotton (AcalaDTL-90) and the Bt cotton (AcalaDTL-90B). The morphological, histochemical, and ultrastructural characteristics of the A. argillacea midgut were very similar to those previously described for most lepidopterans. However, the mitochondrial polymorphism and bifurcate microvilli suggest an ultrastructural and physiological modification, which may cause a higherabsorption and secretion in the columnar cells. Such hypothesis may account for faster toxin and/or microbial agent action, suggesting that this species may be highly susceptible to some ofthese agents. After 20 min, A. argillacea 4th-ínstar larvae feeding on Bt cotton leaves (AcalaDTL-90B) showed alterations on the midgut walls. During that time, morphological and ultrastructural alterations in the goblet and columnar cells were observed, although more prominent on the former. Also, a reduced number of regenerative cells, a deterioration of the muscle layer, and the peritrophic membrane destruction in some midgut regions were observed. The study overall conclusion is that the Cry1Ac toxin acts relatively fast in the A. argillacea midgut, causing severe and irreversible alterations. Alabama argillacea (Hübner) (Lepidoptera: Noctuidae) é a principal praga desfolhadora da cultura do algodão, provocando perdas entre 21 e 35%. Plantas transgênicas que expressam proteínas tóxicas de genes da bactéria Bacillus thuringiensis (Berliner) (Bt) foram recentemente liberadas no Brasil. O algodão Bt AcalaDTL-90B, que expressa a proteína Cry1Ac, é resistente às principais pragas da Ordem Lepidoptera que afetam a cultura do algodão.. Embora as toxinas de Bt interajam com pelo menos um receptor final, desencadeando o efeito tóxico, vários fatores endógenos estão associados à seletividade diferencial das toxinas Bt. Desta forma, estudos da interação da Cry1Ac com o intestino de insetos pragas do algodoeiro são de fundamental importância. Assim, esta pesquisa objetivou descrever histoquimicamente, morfologicamente e ultra-estruturalmente o mesêntero de larvas de A. argillacea alimentadas com a isolinha (AcalaDTL-90), bem como algodão-Bt (AcalaDTL-90B). As características morfológicas, histoquímicas e ultra-estruturais do mesêntero de A argillacea, de modo geral, foram semelhantes às descritas na literatura para a maioria dos insetos da Ordem Lepidoptera. No entanto, opolimorfismo mitocondrial e microvilosidades bifurcadas sugerem uma modificação ultra-estrutural e fisiológica, que pode ocasionar uma maior absorção e secreção nas células colunares. Tal hipótese pode favorecer uma ação mais rápida de toxinas e/ou agentes microbianos,sugerindo que esta espécie pode ser altamente suscetível para alguns destes agentes. Larvas de 4º ínstar após 20 min de alimentação com folhas de algodão Bt (AcalaDTL-90B) mostraram modificações na parede do mesêntero de A. argillacea. Neste tempo de alimentação, foram observadas alterações morfológicas e ultra-estruturais nas células colunares e caliciformes, sendo mais expressivas nessas últimas. Também houve uma redução no número das células regenerativas, degeneração da camada muscular e destruição da membrana peritrófica em algumas regiões do mesêntero. Assim, conclui-se que a toxina Cry1Ac age relativamente rápida no mesêntero da A. argillacea, ocasionando alterações drásticas e irreversíveis.

WOS: 000267788000002, The objective of this investigation was to study morphological aspects of the Harderian, paraorbital, gland in domestic geese. Tissues were obtained from 9 male and 9 female adult domestic geese and processed for light microscopy. Tissue sections were stained with Mallory's trichrome, silver impregnation, methyl green-pyronin, periodic acid Schiff, alcian blue (pH 2.5), alcian blue/periodic acid Schiff (pH 2.5) and Sudan black B staining techniques. Results revealed the Harderian gland of domestic geese was covered by a capsule, which contributed trabeculae that divide the organ into lobes and lobules. The glandular epithelium was multilobular, compound tubuloalveolar. The secretion of Harderian gland was mucous and the secretion type was apocrine; the gland emptied to the inner surface of the nicticating membrane by a single duct.

Iron is a metal of vital importance for many organisms as it plays an essential role in numerous metabolic processes. It is an integral part of various proteins such as haemoglobin, myoglobin, cytochromes as well as of numerous enzymes such as catalases, peroxidases, dehydrogenases (succinate dehydrogenase, mitochondrial NADH dehydrogenase) and enzymes involved in DNA synthesis. Although essential, iron can be toxic when present in excess as it catalyses the generation of wide range of free radicals, including the hydroxyl radical, which can react with a large number of cell components (DNA, proteins, lipids, sugars) and can cause serious cellular damage [1]. The aim of this study was to monitor iron accumulation in carp liver tissue during long-term exposure to maximum allowed waterborne iron concentration (1 mg/L) in the form of iron-gluconate (FeG) and iron-dextran (FeD). Histological changes in liver and the activity of lactate dehydrogenase (LDH), a marker for cell/tissue damage, were also examined [2, 3]. Carp were held for 70 days in aquaria with constant iron concentration (1 mg/L). Liver tissue samples of control and exposed animals were taken every day during the first four days of the treatment and in intervals of five or seven days until the final 70th day of the treatment. Histochemical modified Perl´s reaction was used to detect iron in liver tissue [3]. Iron depositions were observed by light microscope and analyzed with image analyser (LUCIA G 4.81). In carp liver of the control group, Perl´s reaction showed no iron deposits (Fig. 2). Iron started to accumulate on the 28th day of the treatment with both FeG and FeD and deposits were located in the sinusoid endothelium as mildly diffused reactions. Much less was found in the cytoplasm of hepatocytes in the form of grain and diffused reaction. After the 35th day of the treatment, iron was also accumulated in macrophages. Continued treatment with FeG and FeD increased the quantity of iron depositions in liver tissue (Fig. 1, 2). The results indicate that long-term exposure of carp to low iron concentrations causes significant Fe accumulation (p

Submitted by (edna.saturno@ufrpe.br) on 2016-11-23T13:13:53Z No. of bitstreams: 1 Alicely Araujo Correia.pdf: 1178729 bytes, checksum: 007e1cb0c61d3a8f7fdcf0344b043ecf (MD5) Made available in DSpace on 2016-11-23T13:13:53Z (GMT). No. of bitstreams: 1 Alicely Araujo Correia.pdf: 1178729 bytes, checksum: 007e1cb0c61d3a8f7fdcf0344b043ecf (MD5) Previous issue date: 2008-02-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is the main pest of corn crops. The control of S. frugiperda requires various insecticide applications that might result on selection of resistant populations and environmental contamination. A promising alternative toward the control of this pest is the use of vegetal insecticides such as neem, Azadirachta indica A. Juss (Meliaceae). Few data exist related to the immunologic and histophysiologic reactions of S. frugiperda larvae under neem treatment. Thus, the present research had as objectives: (a) to characterize and to quantify the hemocytes of S. frugiperda larvae pre- and post-treatment with concentrations 0.5 and 1.0% of neem (Neemseto®) on the intervals of 48, 96, 144, 192 and 240h; (b) to describe, histologically, the food channel of the larvae; (c) to characterize the effect of the concentrations 0.5 and 1.0% of neem, in the same evaluation intervals after treatment, on the histophysiology of midgut; (d) to quantify the regenerative cells and; (e) to analyze,histochemically, the secretion product of midgut. Six types of hemocytes were identified in the hemolymph of S. frugiperda larvae: Adipohemocytes, Spherulocytes, Granulocytes, Eonocytoids, Plasmatocytes and Prohemocytes. And, the most abundant hemocytes were Granulocytes and Plasmatocytes. The counting of hemocytes showed effects as function of neem concentrations onthe hemocytary dynamics and possivelmente the endocrine events, contributing for mortality and observed morphologic abnormalities. Despite of expressive larval mortality and of effects with neem at concentration 1.0%, both concentrations (0.5 and 1.0%) modified the cellular immunity of the larvae. Histophysiological alterations occurred as function of tested neem concentrations and time after treatment observed as: degeneration of the epithelium and musculature of the midgut, reduction of regenerative cells and the secretory activity of this region, in the two concentrations of neem. These alterations, however, were observed for larvae treated with neem at concentration 1.0% already from 96h after treatment. These results demonstrate the insecticide activity of the commercial formulation neem (Neemseto®) in both tested concentrations, on thehistophysiology of midgut and on the immunologic reactions of S. frugiperda larvae. Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) é a principal praga da cultura do milho. Seu controle demanda elevado número de aplicações de inseticidas sintéticos, podendo ocasionar seleção de populações resistentes e contaminação ambiental. Uma alternativa promissora para o controle deste inseto é a utilização de substâncias com ação inseticida oriundas de plantas como o nim, Azadirachta indica A. Juss (Meliaceae). Contudo, os aspectos relacionados às reações imunológicas e histofisiológicas de lagartas de S. frugiperda, promovidos pela interação com o nim, ainda são pouco estudados. Assim, a presente pesquisa teve como objetivos: (a) caracterizar e realizar a contagem diferencial dos hemócitos de lagartas de S. frugiperda, antes e após o tratamento com nim (Neemseto®) nas concentrações 0,5 e 1,0%, nos intervalos de 48, 96, 144, 192 e 240h; (b )descrever, histologicamente, o canal alimentar dessas lagartas; (c) caracterizar o efeito das concentrações 0,5 e 1,0% do nim, nos mesmos intervalos deavaliação após tratamento, sobre a histofisiologia do mesêntero; (d) quantificar as células regenerativas e; (e) analisar, histoquimicamente, o produto de secreção do mesêntero. Na hemolinfa das lagartas de S. frugiperda foram identificados seis tipos de hemócitos: Adipohemócitos, Esferulócitos, Granulócitos, Oenocitóides, Plasmatócitos e Prohemócitos, sendoos Granulócitos e Plasmatócitos mais freqüentes. A contagem diferencial dos hemócitos revelou um efeito concentração-dependente do nim, afetando a dinâmica hemocitária e possivelmente os eventos endócrinos, contribuindo para a mortalidade e anormalidades morfológicas observadas. Apesar da mortalidade e dos efeitos ocorrerem de forma mais expressiva no tratamento com nim a 1,0%, ambas as concentrações (0,5 e 1,0%) alteraram a imunidade celular das lagartas. Foram observadas alterações histofisiológicas que variaram de intensidade de acordo com o tempo e as concentrações, tais como: degeneração do epitélio e da musculatura do mesêntero, redução de células regenerativas e da atividade secretora desta região, nas duas concentrações do nim. Porém, estas alterações foram observadas nas lagartas tratadas com nim a 1,0% já a partir de 96h após o tratamento. Esses resultados demonstram a atividade inseticida da formulação comercial de nim(Neemseto) em ambas as concentrações testadas, na histofisiologia do mesêntero e nas reações imunológicas de lagartas de S. frugiperda.

8 páginas, 1 figura, 3 tablas. VIII Congreso Nacional de Acuicultura: Acuicultura y desarrollo Sostenible (2001, 22-25 mayo, Santander), [EN] We present a histomorphological description of the digestive tract of the oyster Crassostrea angulata (Lamarck, 1819), as well as a histochemical study of its carbohydrate distribution. The study shows that, in general, glycogen is found in the connective tissue, and neutral mucopolysaccharides/glycoproteins and/or acid mucosubstances in the epithelium and basal cell layers. Glycogen was not detected in the digestive gland, possibly because of its mobilization to the connective tissue for reproduction., [ES] Se realiza una descripción histomorfológica del aparato digestivo del ostión Crassostrea angulata (Lamarck, 1819) y se pone de manifiesto la distribución de carbohidratos mediante técnicas histoquímicas. La pauta general observada es la presencia de glucógeno en el tejido conjuntivo y de mucopolisacáridos/glicoproteínas neutras y/o ácidas en las membranas basales y el epitelio. En la glándula digestiva no se observa glucógeno debido, posiblemente, a su movilización hacia el tejido conjuntivo para su posterior uso con fines reproductivos.

Swine skeletal muscle fibers have traditionally been classified as types I, IIA, IIB and their hybrid forms by determining the activity of myosin ATPase (mATPase). More recently, molecular, electrophoretic and immunohistochemical approaches have allowed the identification of IIx isoform-containing fibers. In this study, myosin isoforms present in different types of muscle fibers were identified by combining histochemical and immunohistochemical methods. Samples from the M. longissimus dorsi were collected from primiparus pigs (mean body weight, 150 Kg) 45 minutes after slaughtering and subsequently frozen in dry ice-cooled acetone. Serial cross sections were processed for mATPase and NADH-TR activities and for immunohistochemistry. Five histochemical fiber groups were identified: type I (dark), type IIA (light) and three intermediate types. A panel of monoclonal antibodies identified the slow ßI, IIa, IIx and IIb isoforms present in types I, IIA, IIX and IIB fibers and the hybrid group IIAX Las fibras musculares esqueléticas del cerdo han sido tradicionalmente tipificadas como I, IIA y IIB, más sus formas híbridas, determinando la actividad de la enzima miosina ATPasa miofibrilar. Recientemente, la utilización de anticuerpos monoclonales contra isoformas de cadena pesada de miosina, electroforesis y estudios moleculares, han documentado la existencia, además, de la isoforma IIx. El objetivo de este estudio fue determinar la existencia de las isoformas de miosina, presentes en los distintos tipos fibrilares, utilizando técnicas histoquímicas e inmunohistoquímicas combinadas, dentro de una unidad experimental compuesta por muestras musculares del M. longíssimo del dorso, en cerdas de 150 kg promedio. Las muestras fueron obtenidas 45 minutos después de la faena, mediante escisión y congeladas en acetona enfriada con hielo seco. Cortes seriados de 10µm de espesor fueron tratados por la técnicas de mATPasa modificada por Nwoye (1982), a pH 4.6 , NADH-TR e inmunohistoquímica. Histoquímicamente fueron identificados cinco grupos de fibras: tipo I oscuras, IIA claras, y tres tipos intermedios. Los ensayos inmunohistoquímicos permitieron identificar las isoformas ß lenta I, IIa, IIx y IIb presentes en fibras de tipo I, IIA, IIX, IIB y un grupo híbrido IIAX

A comparative study of metastic and primitive tumors extra-celular matrix in nude rats, xenotransplanted with KB cells, using histochemical methods, was carried out. In both primitive and metastatic tumors it was possible to observe a qualitative and a quantitative variation of matricial components; coexistence of different kinds of fibers; small representation of elastic fibers; presence of acid and sulfated glycosaminoglycans and neutral polysacharyds, and also absence of basal membrane. It is suggested a small interaction between collagenic components and proteoglycan of primary tumors and the proteoglycan, whereas in métastasés this interaction is increased. Estudou-se comparativamente, através de métodos histoquímicos, a expressão dos componentes da matriz extracelular de tumores primitivos e metastáticos em ratos nude, xenotransplantados com células KB. Em ambas as neoplasias observou-se uma variabilidade tanto qualitativa como quantitativa dos componentes matriciais, coexistência de diferentes tipos de fibras, pouca representatividade de fibras elásticas de glicosaminoglicanas ácidas e sulfatadas e de polissacarídeos neutros, além da ausência de membrana basal.

Soft and firm stem end tissues of cucumbers fermented at a low concentration of sodium chloride were examined by a combination of light microscopy {LM), transmission electron microscopy { TEM), and scanning electron microscopy { SEM) . When compared to firm tissue, softened tissue cell walls were swollen and striated, and the middle lamellae were poorly stained by the uranyl acetate-1 ead citrate used in TEM . When the tissue was penetrated by a circu l ar punch {1.5 mm diameter), cells in soft tissue tore along the middle l arne ll ae rather than across cell walls, as did cells in firm tissue. Thus, softening was characterized by a weakening of cell-to-cell junctions at the middle lamellae. Ultrastructural changes in the softened tissue were consistent with cellulolytic and / or pectinolytic degradation of cell wall components . Since fungal hyphae were found in both soft and firm tissue, it was not possible to determine if the softening was due to extracellular enzymes produced by invading organisms or to enzymes endogenous to the cucumber tissue.

Morphological, anatomical, and histochemical effects of S-ethyl dipropylthiocarbamate (EPTC) on Avena sativa var. overland were studied after treatment of seeds at various EPTC concentrations. The oat seed germination percentage was not affected by EPTC treatment. EPTC delayed initial root and coleoptile development at all concentrations used. Two or three days after treatment, however, the roots of seedlings treated with EPTC concentrations lower than 3 ppm grew at the same rate as the untreated seedlings and showed no abnormalities. Primary and adventitious root growth of seedlings treated with 3 ppm and higher was inhibited. Most of the adventitious roots remained in the radicle stage and failed to elongate. Bases of the roots became necrotic and were quite brittle. Formative effects were greater in the shoot than in the root. As result of treatment, coleoptiles became chlorotic, thickened, and were hard and brittle. The first true leaf of treated seedlings tended to adhere to the coleoptile and was broken as the coleoptile elongated. The complete shoot meristem of 12 ppm EPTC treated seedlings broke at the base. Young leaves formed within coleoptiles of treated seedlings oftain failed to emerge. Histochemical studies showed more carbohydrates in the coleoptiles of the EPTC treated oat seedlings than in those of the untreated seedlings. The lip1d-containing materials in the coleoptiles of the germinating oat seedlings disappeared from the coleoptiles of fully grown untreated oat seedlings, while they still persisted in the coleoptiles of 5-day-old EPTC treated oat seedlings. This suggests that EPTC might inhibit the breakdown and utilization of stored foods.

Bu çalışmada palpebra tersiya (PT)'nin ve sahip olduğu bezlerin histolojik yapılannın aydınlatılması, gözün lokal bağışıklığına muhtemel katklannin değerlendirilmesi ve bu bölgede gerçekleştirilecek olan lokal müdahalelere yardımcı olabilecek histolojik bilgi birikimine katkıda bulunması amaçlanmıştır.Bu amaçla isviçre esmeri ve Holştayn sığır ırkının her iki seksinde, PT'nin histolojik ve histokimyasal özelikleri ışık mikroskopik düzeyde karşılaştırmalı olarak incelendi, PT'deki bezlerin histokimyasal özelliklerinin belirlenmesinde; periyodik asit-Schiff (PAS), alsiyan blue pH=2.5 (AB) ve PAS/AB pH=2.5 kombine boyama metodlan uygulanmıştır. PT'nin her iki sığır arkında, medial göz açısında yerleşmiş olduğu ve hiyalin kakırdak türünde "T" harfi şeklindeki, KPT ile desteklenmiş olan konjunktiva dürümünden oluştuğu belirlendi. PTnin serbest uç.palpebral ve bulbar yüzlerinin,konjunktivada olduğu gibi çok katlı yassi epitelle örtüldüğü ve kıkırdak çevresinde GPTS'nin lopçuklarının yerleşmiş olduktan belirlendi. Her iki sağir irkinda GPTS'den sonra ki bez bölümü GPTP'nin özelliderini taşımakla birlikte; bu bezin Harder bezinin histolojik kriterlerine sahip olmadiği sonucuna vanildi.PT'nin bağ dokusunda yaygın lenfosit infiltrasyonlarının bulunduğu ve bunların bulbar ile palpebral yüzlerinin lamina propriyalanında ve for. niks konjunktiva bölümünde soliter ve agregat lent foliküllerini oluşturduklan saptanmıştır. GPTS sığırlarda bileşik tubuloalveoler yapıda ve mikst karakterde salgı yapan bezlerin histolojik özelliklerini gösterdiği tespit edildi. İncelenen sığır irklarında PT'nin serbest uç palpebral ve bulbar yüz epitellerinde, özellikle epitelin bazal katmanlarında değişen derecelerde pigmentasyon gözlenmiştir. Bu çalışmada,epitel atı bölümleri siki,derin bölümleri gevşek bağ dokusu özelliği gösteren organ lamina propriyasının yer yer elastik ve retikulum ipliklerini de içeren; kollagen ipliklerle de desteklenmiş olduğu sap tanmıştır. Plazma hücrelerinin,organın serbest ucu ile palpebral ve bulbar yüzlerinin lamina propriyalanında ve GPTS ile GPTP'nin intralobuler ve interlobuler intersitisyumlarında genellikle gruplar oluşturdukları tespit edilmiştir. Miyoepitel hüc releri, GPTS ve GPTP'nin KGleri ile akitici kanallarının duvarını oluşturan epitel hücrelerinin bazal yüzlerinde gözlenmiştir., The aim of study was to determine histological structure of both nictitating membrane, itself and, its glands, to evaluate possible contribution to the local immunity of the eye itself and the basic histological knowledge to be used as reference out on local interventions. For these purposes a comparative histochemical and light microscopic evaluation of PT was carried out on the both sexes in Brown Swiss and Holstein cows. The periodic acid-Schiff (PAS),alcian blue pH=2.5 (AB) and PAS/AB at pH-2.5 combined staining was used to histochemical properties in the glands of the PT.It was coserved that the PT of cows investigated had an T shaped hyaline cartilage, so-called cartilago palpebra tertia (KPT) which was covered with the continuation of conjunctival epithelial tissue in the medial canthus in cows. The epithelia of the both palpebral and bulbar faces, and free marginal aspect of the PT were stratified squamous epithelium as in conjunctiva. Gl palpebra tertia superficialis (GPTS) was observed as lobes surrounded by connective tissue capsule located around the KPT Although the gl. palpebra tertia profundus (GPTP) were observed in the cow, it was lack of the histological and his tochemical properties of the Harderian gland. The cows investigated wide lymphocytic infiltration areas were observed in the connective tissue Especially, cows the areas formed lymphoid follicles in the fomix conjunctiva and lamina propria of the palpebral aspects of the PT.Branched tubuloalveolar construction of GPTS with mix secretion was found in cows Varying degrees of pigmentation was observed over the free margin palpebral and bulbar aspects of the PT in cows Elastic reticular and collagen fibers were observed under the epitheliumn and deeper parts of the lamina propna Plasma cells were found in groups in the connective tissue of the free margin palpebral and bulbar aspects, and int ralobular and interiobular interstitium of the GPTS and GPTP Myoepithelial cells were found to be located in the corpus glandulae and walls of the ducts systems of both GPTS and GPTP.

Made available in DSpace on 2013-08-12T18:37:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-01-01 Made available in DSpace on 2013-09-30T19:12:04Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-01-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:30:35Z No. of bitstreams: 0 Made available in DSpace on 2014-05-20T15:30:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-01-01 This study investigated if overfed rats present morphological and histochemical muscle adaptation similar to normally fed, both submitted to two different weekly frequencies of training. Thirty male Wistar rats were fed either with standard chow (SCO) or with hypercaloric diet (HCO). They were subdivided into six subgroups: sedentary (SCO and HCO), trained twice/week (SC2 and HC2) and trained five times/week (SC5 and HC5). The trained groups swam 60 min/day, during 10 weeks. Twenty four hours after the last training, samples of Gastrocnemius were excised and stained with HE, NADH-TR and m-ATPase, and the capillary density was calculated. Total heart mass (HM) and the mass of atrium (AM), left (LV) and right (RV) ventricles were excised and weighted. The comparisons were made by ANOVA and by Covariance analysis, adjusting the variables by body weight. The results showed that the HCO achieved higher BM, however, absolute HM did not differ post training. Irrespective of the diet, rats that were trained twice a week presented significantly greater increase in the AM. In general, the SC5 and HC5 groups showed higher HM, LV, RV, proportion of oxidative fibres and capillary density, compared to the sedentary and twice week trained groups. A higher proportion of injuries (splitting) was noted in the HC2 and HC5 compared to SC2 and SC5. These results indicate that the frequency of training influenced the skeletal and heart adaptation and larger changes were observed in the 5x/week group, which ingested the standard diet. The 5x/week training groups also presented large amount of muscle fibres damage. São Paulo State Univ, UNESP, Dept Phys Educ, São Paulo, Brazil São Paulo State Univ, UNESP, Dept Phys Educ, São Paulo, Brazil