Efeito da sedação na microcirculação de pacientes em choque séptico

Ao longo dos últimos anos, apesar de todo desenvolvimento e pesquisa, a mortalidade na sepse permanece elevada. Na área de microcirculação foram realizados estudos em modelos experimentais de sepse ao longo das últimas duas décadas, quando se observou, através de técnicas invasivas, alterações como redução expressiva da densidade capilar funcional. A técnica denominada sidestream dark field (SDF) imaging, recentemente desenvolvida, permite a avaliação da microcirculação de forma transcutânea. A utilização desta técnica permitiu evidenciar a redução da densidade capilar funcional em pacientes com sepse grave quando comparado a um indivíduo saudável. Posteriormente, foi demonstrado que alterações persistentes na microcirculação de pacientes sépticos, mesmo com sinais vitais estabilizados, estão associadas com pior prognóstico.Evidentemente, os pacientes com sepse grave ou choque séptico sofrem uma grande quantidade de intervenções terapêuticas, aonde muitas delas alteram a microcirculação. Estudos analisando a microcirculação em pacientes em uso de nitroglicerina, corticóide, recebendo hemotransfusão ou ainda infusão de noradrenalina foram publicados recentemente.Entretanto, até o presente momento, não existem publicações que descrevam a influência dos sedativos na microcirculação de pacientes com choque séptico. As drogas mais comumente utilizadas para sedação de pacientes em ventilação mecânica são o sedativo midazolam e o anestésico propofol. Os objetivos do estudo foram: avaliar o efeito dos principais agentes sedativos utilizados na prática clínica na microcirculação de pacientes com choque séptico utilizando a técnica de sidestream dark field imaging, comparar os efeitos na microcirculação do midazolam com o propofol em pacientes com choque séptico e verificar se existe relação das alterações microcirculatórias provocadas pelos sedativos com as variações de diferentes parâmetros hemodinâmicos, gasométricos ou metabólicos como pressão arterial, índice cardíaco, lactato e saturação venosa central de oxigênio. Foram estudados (estudo prospectivo) 16 pacientes internados no Centro de Terapia Intensiva da Casa de Saúde São José. Os pacientes internados com diagnóstico de choque séptico e que possuíam indicação clínica de ventilação mecânica e de suspensão diária da sedação foram submetidos ao estudo da microcirculação na mucosa sublingual utilizando a técnica de sidestream dark field imaging. Estes pacientes foram sedados conforme orientação do protocolo já existente de sedação, inicialmente com propofol e posteriormente com midazolam. Os principais resultados observados foram:a macrohemodinâmica não diferiu nos 2 momentos do exame, o BIS (bispectral índex of sedation) se manteve na faixa recomendada nos 2 momentos do exame, tendo aumentado quando o paciente acordava, conforme esperado, e a proporção de vasos pequenos perfundidos e o índice de fluxo da microcirculação foram significativamente menores, enquanto o índice de heterogeneidade foi significativamente maior quando os pacientes estavam recebendo infusão de propofol quando comparados com a infusão de midazolam. Concluímos que, em pacientes com choque séptico, a administração de midazolam resulta em uma melhora dos parâmetros microcirculatórios quando comparada com a administração de propofol. Essa diferença não pode ser atribuída a alterações de variáveis hemodinâmicas sistêmicas.

Capacidade de invasão, persistência intracelular e atividade citotóxica de cepas de Aeromonas spp. em modelo celular in vitro e ex vivo

As Aeromonas são consideradas patógenos em potenciais para o homem e animais e estão amplamente distribuídas no ambiente sendo a água e os alimentos importantes veículos de transmissão. Muitos estudos têm demonstrado que a patologia causada pela infecção por Aeromonas é complexa e envolvem inúmeros fatores de virulência, dentre eles a aderência, invasão, enterotoxinas, hemolisinas, exoenzimas, sideróforos, flagelos, formação de biofilme e mecanismos de secreção. No presente estudo, analisamos os mecanismos de patogênese mediados por A. caviae e A. hydrophila, avaliando a participação desses microrganismos nos processos de adesão, invasão, persistência intracelular e citotoxidade celular. Foram utilizados ensaios quantitativos in vitro para testar associação, invasão e persistência intracelular em linhagens celulares HEp-2 e/ou T84. A interação de tecidos intestinais de coelho cultivados in vitro (IVOC) com três cepas de A. caviae originárias de fezes diarréicas também foi avaliada. Observamos que 10 (62,5%) das 16 cepas de Aeromonas spp. de diferentes origens, submetidas aos testes de invasão quantitativos foram capazes de invadir células HEp-2 e T84 em 6 horas de incubação. As cepas positivas nos testes de invasão foram submetidas ao teste quantitativo de persistência em células HEp-2 e sobreviveram no ambiente intracelular por 48 e/ou 72 horas sem multiplicação. A interação de três cepas de A. caviae com a mucosa intestinal de coelho ex vivo resultou em aderência, produção de muco e alterações como, intensa vacuolização e drástica desorganização estrutural que levaram a destruição das microvilosidades intestinais. Este estudo demonstrou que subconjuntos de cepas de A. caviae e A. hydrophila de diversas origens, foram capazes de invadir, persistir ou destruir linhagens celulares in vitro. Nosso estudo também evidenciou que cepas de A. caviae causaram expressivas alterações morfológicas que resultaram na destruição de epitélios intestinais de coelho ex vivo. Finalmente, nossos resultados contribuíram para reforçar o potencial patogênico de cepas de Aeromonas, em especial, as de origem vegetal e clínica.

Age and growth of the estuarine dolphin (Sotalia guianensis) (Cetacea, Delphinidae) on the Paraná Coast, southern Brazil

Teeth of 71 estuarine dolphins (Sotalia guianensis) incidentally caught on the coast of Paraná State, southern Brazil, were used to estimate age. The oldest male and female dolphins were 29 and 30 years, respectively. The mean distance from the neonatal line to the end of the first growth layer group (GLG) was 622.4 ±19.1 μm (n=48). One or two accessory layers were observed between the neonatal line and the end of the first GLG. One of the accessory layers, which was not always present, was located at a mean of 248.9 ±32.6 μm (n=25) from the neonatal line, and its interpretation remains uncertain.The other layer, located at a mean of 419.6 ±44.6 μm (n=54) from the neonatal line, was always present and was first observed between 6.7 and 10.3 months of age. This accessory layer could be a record of weaning in this dolphin. Although no differences in age estimates were observed between teeth sectioned in the anterior-posterior and buccal-lingual planes, we recommend sectioning the teeth in the buccal-lingual plane in order to obtain on-center sections more easily. We also recommend not using teeth from the most anterior part of the mandibles for age estimation. The number of GLGs counted in those teeth was 50% less than the number of GLGs counted in the teeth from the median part of the mandible of the same animal. Although no significant difference (P>0.05) was found between the total lengths of adult male and female estuarine dolphins, we observed that males exhibited a second growth spurt around five years of age. This growth spurt would require that separate growth curves be calculated for the sexes. The asymptotic length (TL∞), k, and t0 obtained by the von Bertalanffy growth model were 177.3 cm, 0.66, and –1.23, respectively, for females and 159.6 cm, 2.02, and –0.38, respectively, for males up to five years, and 186.4 cm, 0.53 and –1.40, respectively, for males older than five years. The total weight (TW)/total length (TL) equations obtained for male and female estuarine dolphins were TW = 3.156 × 10−6 × TL 3.2836 (r=0.96), and TW = 8.974 × 10−5 × TL 2.6182 (r=0.95), respectively.

Amino acid requirements for maturation of rhesus monkey (Macacca mulatta) oocytes in culture

This study evaluated the effects of different amino acid formulations on supporting meiotic and cytoplasmic maturation of rhesus monkey (Macacca mulatta) oocytes in vitro. Five hundred and forty-six cumulus-oocyte complexes (COCs) aspirated from unstimulated adult monkey follicles (greater than or equal to 1000 mum in diameter) were cultured in either modified Connaught Medical Research Laboratories 1066 medium (mCMRL-1066) or in one of eight chemically defined media (modified basic medium 5 supplemented with 5.5 mmol glucose l(-1), 0.003 mmol pantothenic acid l(-1) and different amino acid formulations) as below: (1) modified basic medium 5 (mBM5) containing no amino acid; (2) mBM5 + 0.2 mmol glutamine l(-1); (3) mBM5 + 11 amino acids from hamster embryo culture medium 6 (HECM-6) (11 AA); (4) mBM5 + Eagle's non-essential amino acids (NEA); (5) mBM5 + NEA + 0.2 mmol glutamine l(-1); (6) mBM5 + Eagle's essential amino acids (EA) without glutamine; (7) mBM5 + EA + 0.2 mmol glutamine l(-1); (8) mBM5 + Eagle's 20 amino acids (20 AA) + 0.2 mmol glutamine l(-1); and (9) mCMRL-1066 (control). All media contained FSH, LH, oestradiol and progesterone. After maturation, mature oocytes were subjected to the same fertilization and embryo culture procedures. COCs matured in treatment 5 had greater potential to progress to metaphase II (66%; P < 0.05) than did those in treatments 1 (37.3%), 2 (48.3%)f 3 (41%), 6 (41%) and 9 (43%). Oocytes matured in treatment 8 had the best morula (53%) and blastocyst (18%) developmental responses (P<0.05). The lowest (P<0.05) morula and blastocyst developmental responses were obtained from COCs matured in treatments 1 (0%) and 6 (8%). The other media supported intermediate embryonic development (range 11-38% of morula and blastocyst). These results indicate that the choice of amino acids affects the competence of oocyte maturation and that Eagle's 20 AA with 0.2 mmol glutamine l(-1) is more efficient than the other amino acid formulations for maturation of rhesus monkey oocytes.

Impact of phosphamidon and its metabolites on histopathology of the liver, gill and intestine of Labeo rohita

Impact of phosphamidon, an organophosphorus pesticide and its metabolites viz. dimethyl phosphoric acid and 2-chloro 2-diethyl carbamoylmethyl vinyl acid on histopathology of a common teleost, Labeo rohita was studied by exposing the fish to sub-lethal concentrations which were taken as 1/3rd of LC50 and were equal to 0.0123 ppm for phosphamidon, 0.0160 ppm for dimethyl phosphoric acid and 0.0167 ppm for 2-chloro 2-diethyl carbamoylmethyl vinyl acid respectively. The results revealed that hepatocytes in the liver were markedly swollen and exhibited hydropic degeneration. Fusion of primary lamellae and moderate congestion of blood vessels were evident in the gill. Intestine showed degeneration of mucosa and cellular infiltration in sub-mucosa. LC50 values and histopathological photomicrographs suggest that phosphamidon is more toxic as compared to dimethyl phosphoric and 2-chloro 2-diethyl carbamoylmethyl vinyl acid.

Redescription of Camallanus hypophthalmichthys Dogel and Akhmerov, 1959 (Nematoda : Camallanidae) and its first record from fishes in China

The nematode Camallanus hypophthalmichthys Dogel and Akhmerov, 1959 is redescribed from specimens collected from the intestine of the bighead carp Aristichthys nobilis, from Liangzihu Lake (Yangtze River basin), Hubei Province, central China. The light and scanning electron microscopical examination made it possible to study in detail the morphology of this so far little-known species and to confirm its validity. The main specific features of C. hypophthalmichthys distinguishing it from the most similar Camallanus spp. is the presence of 3 small caudal processes on the male tail tip, 13-16 longitudinal ridges on the inner surface of the valve of the buccal capsule, and the arrangement of preanal and postanal genital papillae in the male. This finding represents a new host record, the first record of this parasite in the Yangtze River basin, and the first documented record of C. hypophthalmichthys from China. Camallanus hypophthalmichthys is considered a specific intestinal parasite of fishes of the cyprinid Hypophthalmichthyinae.

Total-internal-reflection fluorescence microscopy with W-shaped axicon mirrors

A scheme based on a W-shaped axicon mirror device for total-internal-reflection fluorescence microscopy (TIRFM) is presented. This approach combines the advantages of higher efficiency compared with traditional TIRFM, adjustable illumination area, and simple switching between wide-field and TIRF imaging modes. TIRF images obtained with this approach are free of shadow artifacts and of interference fringes. Example micrographs of fluorescently labeled polystyrene beads, of Convallaria majalis tissue, and of Propidium-iodide-labeled Chinese hamster ovary cells are shown, and the capabilities of the scheme are discussed. (C) 2010 Optical Society of America

Protective effects of melatonin against low- and high-LET irradiation

To investigate the protective effects of melatonin against high-LET ionizing radiation, V79 Chinese hamster cells were irradiated with 100 keV/mu m carbon beam. Parallel experiments were performed with 200 kV X-rays. To avoid the impact from extra solvents, melatonin was dissolved directly in culture medium. Cells were cultured in melatonin medium for 1 hr before irradiation. Cell inactivation was measured with conventional colony forming assay, medium containing 6-thioguanine was used for the selection of mutants at hprt locus, and the cell cycle was monitored by flow cytometry. Both carbon beam and X-rays induced cell inactivation, hprt gene mutation and cell cycle G2 block dose-dependently. But carbon beam showed stronger effects as indicated by all three endpoints and the relative biological effectiveness (RBE) was 3.5 for cell killing (at 10% survival level) and 2.9 for mutation induction (at 5 x 10(-5) mutants/ cell level). Melatonin showed protective effects against ionizing radiation in a dose-dependent manner. In terms of cell killing, melatonin only increased the survival level of those samples exposed to 8Gy or larger of X-rays or 6 Gy or larger of carbon beam. In the induction of hprt mutation and G2 block, melatonin reduced such effects induced by carbon beam but not by X-rays. The results suggest that melatonin reduces the direct interaction of particles with cells rather than an indirect interaction. Further studies are required to disclose the underlying mechanisms.

Multi-armed poly(L-glutamic acid)-graft-oligoethylenimine copolymers as efficient nonviral gene delivery vectors

Background The application of polyethylenimine (PEI) in gene delivery has been severely limited by significant cytotoxicity that results from a nondegradable methylene backbone and high cationic charge density. It is therefore necessary to develop novel biodegradable PEI derivates for low-toxic, highly efficient gene delivery.Methods A series of novel cationic copolymers with various charge density were designed and synthesized by grafting different kinds of oligoethylenimine (OEI) onto a determinate multi-armed poly(L-glutamic acid) backbone. The molecular structures of multi-armed poly(L-glutamic acid)-graft-OEI (MP-g-OEI) copolymers were characterized using nuclear magnetic resonance, viscosimetry and gel permeation chromatography. Moreover, the MP-g-OEI/DNA complexes were measured by a gel retardation assay, dynamic light scattering and atomic force microscopy to determine DNA binding ability, particle size, zeta potential, complex formation and shape, respectively. MP-g-OEI copolymers were also evaluated in Chinese hamster ovary and human embryonic kidney-293 cells for their cytotoxicity and transfection efficiency.

Expression and localization of a novel phosducin-like protein from amphioxus Branchiostoma belcheri

A full length amphioxus cDNA, encoding a novel phosducin-like protein (Amphi-PhLP), was identified for the first time from the gut cDNA library of Branchiostoma belcheri. It is comprised of 1 550 bp and an open reading frame (ORF) of 241 amino acids, with a predicted molecular mass of approximately 28 kDa. In situ hybridization histochemistry revealed a tissue-specific expression pattern of Amphi-PhLP with the high levels in the ovary, and at a lower level in the hind gut and testis, hepatic caecum, gill, endostyle, and epipharyngeal groove, while it was absent in the muscle, neural tube and notochord. In the Chinese Hamster Ovary (CHO) cells transfected with the expression plasmid pEGFP-N1/Amphi-PhLP, the fusion protein was targeted in the cytoplasm of CHO cells, suggesting that Amphi-PhLP is a cytosolic protein. This work may provide a framework for further understanding of the physiological function of Amphi-PhLP in B. belcheri.

A new spirofilid ciliate from the yellow sea: Metastrongylidium distichum n gen, n sp (ciliophora, stichotrichia)

The morphology and infraciliature of a new ciliate, Metastrongylidium distichum, isolated from the Yellow Sea, are investigated using live observation and protargol impregnation. Metastrongylidium distichum is about 170 x 40 pm in vivo, clavate to elongate ellipsoidal with bluntly pointed posterior end, and has two macronuclear nodules, six distinctly large buccal and frontal cirri, three dorsal kineties and two each of spiralled ventral and marginal cirral rows. These features indicate a generic allocation in Strongylidium Sterki, 1878. However, the new ciliate has a distinct feature not recognizable in Strongylidiurn, viz., the presence of (three or four) transverse cirri. Thus, we propose a new genus Metastrongylidium for the new species, M distichum nov. gen., n. sp. Metastrongylidium belongs to the family Spirofilidae, where it differs from Mucotrichidium by the lack of postperistomial cirrus and the different frontal and ventral cirral pattern. Metastrongylidium distichum is easily distinguishable from the seemingly similar species Strongylidium californicum Kahl, 1932 by the macronuclear pattern (invariably 2 vs. many nodules). It highly resembles the poorly known species S. contortum (Gelei 1954) Borror, 1972 in the body outline and nuclear pattern, differing in the biotope, the posterior cirral pattern, and the arrangement of right marginal row.

Kiitricha minuta n. Sp., a peculiar hypotrichous ciliate (Ciliophora, Spirotrichea) from the Yellow Sea

The morphology and infraciliature of a new ciliate, Kiitricha minuta n. sp., isolated from the Yellow Sea, were investigated using live observation and protargol impregnation. Kiitricha minuta represents a third member of the rarely known order Kiitrichida. It is unique in the subclass Hypotrichia in having many rows of small uniform cirri along the right side of the body and the dorsal kineties composed of dikinetids, most of which bear two cilia. Kiitricha minuta n. sp. is ovoid and measures about 60 x 45 mu m in vivo. It has a huge buccal cavity occupying about 80% of the body length, numerous body extrusomes, one macronucleus and two micronuclei, 27-27 adoral membranelles, 9-12 frontoventral cirral rows, a submarginal row of 7-9 cirri, 6 or 7 transverse cirri, and roughly 7-9 dorsal kineties. This new species differs distinctly from its only congener Kiitricha marina by its smaller size (60 mu m vs. 80-150 mu m), the presence of body extrusomes (vs. absent), the different macronuclear pattern (one vs. two nodules), and the lower number of frontoventral cirral rows (9-12 vs. 21-26), which terminate at the anterior two-thirds of body (vs. extend to the posterior). The new term "submarginal cirral row" is introduced to distinguish from the marginal cirral row in typical hypotrichs sensu lato. Based on our new observations and the literature, an improved diagnosis for the genus Kiitricha is provided and its phylogenetic importance is discussed.

Conservação de recursos fitogenéticos da Amazônia em coleções diversificadas: avaliação do estabelecimento de doze espécies.

O CPAA esta testando um metodo de conservacao fundamentado em aspectos genetico-ecologicos. Em uma area de 0,25ha, procurou-se representar as estrategias utilizadas pelas especies autoctones para manterem-se em equilibrio no ecossistema tropical: diversidade, densidade e variabilidade genetica. A pesquisa, iniciada em janeiro de 1994, estuda a conservacao de recursos geneticos de sete especies frutiferas (Theobroma grandiflorum, T. cacao, Bactris gasipaes, Euterpe spp, Myrciaria dubia, Rollinia mucosa e Couma utilis) e cinco florestais (Hevea spp, Ceiba pentandra, Jacaranda copaia, Buchenavia huber, Trattinickia burserifolia), utilizando-se tres espacamentos. As caracteristicas biometricas das plantas sao anualmente medidas e, apos cinco anos, a viabilidade tecnica sera avaliada quanto a sobrevivencia e capacidade das especies em deixar descendentes. Preliminarmente, as especies que apresentaram valores superiores na avaliacao de estabelecimento foram C. pentandra entre as florestais (medias de 4,76m de altura e 11,46cm de diametro, a 50cm do solo) e R. mucosa entre as fruteiras (media de 3,71m de altura e 6,28cm de diametro, a 50cm do solo).

Sinusite odontogénica

Monografia apresentada à Universidade Fernando Pessoa para obtenção do grau de Licenciada em Medicina Dentária.

Relação entre a Infeção por Escherichia coli Aderente-Invasina e a Doença de Crohn

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas