480 research outputs found

    The evolutionary dynamics of the Helena retrotransposon revealed by sequenced Drosophila genomes

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    <p>Abstract</p> <p>Background</p> <p>Several studies have shown that genomes contain a mixture of transposable elements, some of which are still active and others ancient relics that have degenerated. This is true for the non-LTR retrotransposon <it>Helena</it>, of which only degenerate sequences have been shown to be present in some species (<it>Drosophila melanogaster</it>), whereas putatively active sequences are present in others (<it>D. simulans</it>). Combining experimental and population analyses with the sequence analysis of the 12 <it>Drosophila </it>genomes, we have investigated the evolution of <it>Helena</it>, and propose a possible scenario for the evolution of this element.</p> <p>Results</p> <p>We show that six species of <it>Drosophila </it>have the <it>Helena </it>transposable element at different stages of its evolution. The copy number is highly variable among these species, but most of them are truncated at the 5' ends and also harbor several internal deletions and insertions suggesting that they are inactive in all species, except in <it>D. mojavensis </it>in which quantitative RT-PCR experiments have identified a putative active copy.</p> <p>Conclusion</p> <p>Our data suggest that <it>Helena </it>was present in the common ancestor of the <it>Drosophila </it>genus, which has been vertically transmitted to the derived lineages, but that it has been lost in some of them. The wide variation in copy number and sequence degeneration in the different species suggest that the evolutionary dynamics of <it>Helena </it>depends on the genomic environment of the host species.</p

    The use of immunohistochemistry for diagnosis of prostate cancer

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    PURPOSE: Atypical glands (ASAP) are diagnosed in 5.0% of prostate biopsies, and cancer identification in a rebiopsy is higher than 40.0%. The use of antibodies to mark basal cells is currently a common practice, in order to avoid rebiopsies. There has been no reported study that has reviewed characteristics of radical prostatectomies (RPs) when immunohistochemistry (IHC) was necessary for definitive diagnosis. MATERIALS AND METHODS: Out of 4127 biopsies examined from 2004 to 2008, 144 (3.5%) were diagnosed with ASAP. IHC was performed using antibody anti-34ßE12 and p63. The results of surgical specimens of 27 patients treated by RP after the diagnosis of prostate cancer (PC) was made using IHC (Group 1) were compared with 1040 patients where IHC was not necessary (Group 2). RESULTS: IHC helped to diagnose PC in 103 patients (71.5%). Twenty-seven (26.2%) underwent RP. In Group 1, two (7.4%) adenocarcinomas were insignificant versus 29 (2.9%) for Group 2. Patients from Group 1 were younger (p = 0.039), had lower Gleason scores (GS) (p < 0.001), lower percentage of Gleason pattern 4 (p < 0.001), and smaller tumors (p < 0.001). CONCLUSION: The use of IHC did not lead to diagnosis of insignificant tumors as illustrated by absence of differences in pathological stage or positive surgical margins in men submitted to RP. Therefore, our results suggest that this modality should be routinely used for a borderline biopsy and ASAP cases

    An assay for secologanin in plant tissues based on enzymatic conversion into strictosidine

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    The secoiridoid glucoside secologanin is the terpenoid building block in the biosynthesis of terpenoid indole alkaloids. A method for its determination in plant tissues and in cell suspension cultures has been developed. This assay is based on the condensation of secologanin with tryptamine, yielding strictosidine, in a reaction catalysed by the enzyme strictosidine synthase (STR; E.C. 4.3.3.2). Subsequently, the formation of strictosidine is quantified by high performance liquid chromatography (HPLC). STR was isolated from transgenic Nicotiana tabacum cells expressing a cDNA-derived gene coding for STR from Catharanthus roseus. The high specificity of STR for secologanin, in combination with a sensitive and selective HPLC system, allows a simple extraction of secologanin from plant tissue. The detection limit of this method is 15 ng secologanin. Using this assay, secologanin contents were determined in tissues of various plant species; Lonicera xylosteum hairy roots were found to contain 1% of secologanin on a dry weight basis. # 1998 John Wiley & Sons, Ltd.info:eu-repo/semantics/publishedVersio

    CARACTERIZAÇÃO E AVALIAÇÃO SENSORIAL DE SORVETE COM EXTRATO AQUOSO DE YACON

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    O yacon está sendo considerado um alimento nutracêutico em decorrência dos estudos sobre a diminuição dos níveis de açúcar no sangue após seu consumo repetido por de ser rico em fibras alimentares, especialmente as solúveis ou viscosas, dentre elas inulina e Frutoligossacarídeos (FOS). O crescente aumento da busca no mercado por produtos que apresentem, além de qualidade sensorial e nutricional, benefícios associados à saúde faz com que as pesquisas nesse ramo cresçam. Com isso, o objetivo do trabalho foi elaborar e avaliar um sorvete com extrato aquoso de yacon em diferentes concentrações. Foram elaboradas duas amostras (F1 com adição de 5% de extrato de yacon e F2 com 10%) e realizaram-se análises de composição centesimal (umidade, cinzas, proteínas, lipídeos e carboidratos) e sensoriais (escala hedônica para cor, sabor, textura, aroma e aspecto global), cor e textura instrumental. A composição centesimal não apresentou diferença significativa (p>0,05) para umidade, cinzas e carboidratos. O teor de proteína foi menor para F2 e o teor de lipídeo foi maior para F2. As amostras apresentaram a cor amarela clara com alta luminosidade e não diferiram significativamente na textura expressa em força de compressão. As duas formulações foram sensorialmente aceitas, não apresentando diferença significativa entre si e conferindo um alto índice de aceitação.AbstractYacon is being considered a nutraceutical food due to studies about the decrease of sugar levels in blood after its repeated consumption, and also for being rich in food fibers, specially the soluble or viscous, among them fructooligosaccharides and inulin. The increasing search in the market for products presenting beyond sensory qualities, nutritional and health benefits, makes the researches in this field grows. The goal of this task was to develop an ice cream with aqueous extract of yacon, in different concentrations. It were prepared two samples, (F1 with addition of 5% of yacon extract, and F2 with 10% of it) and were performed centesimal composition analysis (humidity, ashes, protein, lipids, carbohydrates) and sensory analysis (Hedonic scale for color, taste, texture, flavor and global aspect), beyond color and instrumental texture. The centesimal composition showed no significant difference (p> 0.05) for moisture, ash and carbohydrates. The protein content was lower for F2 and lipid content was higher for F2.Samples displayed a clear yellow coloration, with high luminosity and didn’t differ significantly in texture, expressed in compression force. The two formulations were sensory acceptable, showing no significant difference between it and confirming a high acceptance rate

    Saccharomyces cerevisiae como agente probiótico e possível adsorvente de aflatoxina B1 em condições simuladas do trato intestinal de peixes

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    The aim of this study was to evaluate in vitro the probiotic potential and absorption of Saccharomyces cerevisiae for the aflatoxin B1 in simulated fish intestinal tract conditions. Three yeast strains were used, two from brewery: S. cerevisiae RC1 and S. cerevisiae RC3 and one from a fish farming environment: S. cerevisiae A8L2. The selected yeasts were subjected to the following in vitro tests: homologous inhibition, self-aggregation, co-aggregation, antibacterial activity, gastrointestinal conditions tolerance and adsorption of AFB1. All S. cerevisiae strains showed good capability of self-aggregation and co-aggregation with pathogenic bacteria. All yeast strains were able to survive the gastrointestinal conditions. In acidic conditions, the factors (strain vs. time) had interaction (P=0.0317), resulting in significant variation among the strains tested in the time periods analyzed. It was observed that there was also interaction (P=0.0062) in intestinal conditions, with an increased number of cells in the 12-hour period for all strains tested. In the adsorption test, the A8L2 strain was statistically more effective (P<0.005) for both AFB1 concentrations evaluated in this study (10 and 25ng/mL). Thus, it was observed that the strains of S. cerevisiae have potential probiotic and adsorbent of AFB1.Objetivou-se, com esta pesquisa, avaliar in vitro o potencial probiótico e adsorvente de Saccharomyces cerevisiae para aflatoxina B 1 em condições simuladas do trato intestinal de peixes. Foram utilizadas três cepas de leveduras, sendo duas provenientes de cervejaria: S. cerevisiae RC1 e S. cerevisiae RC3, e uma de ambiente de piscicultura: S. cerevisiae A8L2. As leveduras selecionadas foram submetidas aos seguintes testes in vitro: inibição homóloga, autoagregação, coagregação, atividade antibacteriana, viabilidade às condições gastrointestinais e adsorção de AFB 1 . Todas as estirpes de S. cerevisiae mostraram boa capacidade de autoagregação e coagregação com bactérias patogênicas. Todas as estirpes de levedura foram capazes de sobreviver às condições gastrointestinais. Em condições ácidas, os fatores (cepa x tempo) tiveram interação (P=0,0317), resultando em variações significativas entre as cepas testadas nos períodos de tempo analisados. Observou-se que também houve interação (P=0,0062) em condições intestinais, havendo um aumento do número de células no período de 12h para todas as cepas avaliadas. No ensaio de adsorção, a estirpe A8L2 foi a mais eficaz estatisticamente (P<0,005), para as duas concentrações de AFB 1 avaliadas neste estudo (10 e 25ng. mL -1 ). Dessa forma, conclui-se que as cepas de Saccharomyces cerevisiae possuem potencial probiótico e adsorvente de AFB 1.Fil: Pinheiro, R. E. E.. Universidade Federal Do Piaui.; BrasilFil: Rodrigues de Campos, Ana Maria. Universidade Federal Do Piaui.; BrasilFil: Lima, Carlos Eduardo. Universidade Federal Do Piaui.; BrasilFil: Santos, J. T. O.. Universidade Federal Do Piaui.; BrasilFil: Pereyra, Carina Maricel. Universidad Nacional de Río Cuarto. Instituto para el Desarrollo Agroindustrial y de la Salud. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto para el Desarrollo Agroindustrial y de la Salud; ArgentinaFil: Torres, Adriana Mabel. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; ArgentinaFil: Cavaglieri, Lilia Reneé. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; ArgentinaFil: Lopes, J. B.. Universidade Federal Do Piaui.; BrasilFil: Muratori, M. C. S.. Universidade Federal Do Piaui.; Brasi

    Cellular prion protein interaction with vitronectin supports axonal growth and is compensated by integrins

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    The physiological functions of the cellular prion protein, PrPC, as a cell surface pleiotropic receptor are under debate. We report that PrPC interacts with vitronectin but not with fibronectin or collagen. the binding sites mediating this PrPC-vitronectin interaction were mapped to residues 105-119 of PrPC and the residues 307-320 of vitronectin. the two proteins were co-localized in embryonic dorsal root ganglia from wild-type mice. Vitronectin addition to cultured dorsal root ganglia induced axonal growth, which could be mimicked by vitronectin peptide 307-320 and abrogated by anti-PrPC antibodies. Full-length vitronectin, but not the vitronectin peptide 307-320, induced axonal growth of dorsal root neurons from two strains of PrPC-null mice. Functional assays demonstrated that relative to wild-type cells, PrPC-null dorsal root neurons were more responsive to the Arg-Gly-Asp peptide (an integrin-binding site), and exhibited greater alpha v beta 3 activity. Our findings indicate that PrPC plays an important role in axonal growth, and this function may be rescued in PrPC-knockout animals by integrin compensatory mechanisms.Hosp Alemao Oswaldo Cruz, Ludwig Inst Canc Res, São Paulo, BrazilUniv São Paulo, Inst Quim, Dept Bioquim, BR-05508 São Paulo, BrazilHosp Canc, Ctr Tratamento & Pesquisa, São Paulo, BrazilUniv Fed Parana, Dept Patol Basica, BR-80060000 Curitiba, Parana, BrazilUniv Fed Parana, Dept Biol Celular, BR-80060000 Curitiba, Parana, BrazilUniversidade Federal de São Paulo, INFAR, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, INFAR, BR-04023062 São Paulo, BrazilWeb of Scienc

    A Treatment with a Protease Inhibitor Recombinant from the Cattle Tick (Rhipicephalus Boophilus microplus) Ameliorates Emphysema in Mice

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    Aims: To determine whether a serine protease inhibitor treatment can prevent or minimize emphysema in mice.Methods: C57BL/6 mice were subjected to porcine pancreatic elastase (PPE) nasal instillation to induce emphysema and were treated with a serine protease inhibitor (rBmTI-A) before (Protocol 1) and after (Protocol 2) emphysema development. in both protocols, we evaluated lung function to evaluate the airway resistance (Raw), tissue damping (Gtis) and tissue elastance (Htis). the inflammatory profile was analyzed in the bronchoalveolar lavage (BALF) and through the use of morphometry; we measured the mean linear intercept (Lm) (to verify alveolar enlargement), the volume proportion of collagen and elastic fibers, and the numbers of macrophages and metalloprotease 12 (MMP-12) positive cells in the parenchyma. We showed that at both time points, even after the emphysema was established, the rBmTI-A treatment was sufficient to reverse the loss of elastic recoil measured by Htis, the alveolar enlargement and the increase in the total number of cells in the BALF, with a primary decrease in the number of macrophages. Although, the treatment did not control the increase in macrophages in the lung parenchyma, it was sufficient to decrease the number of positive cells for MMP-12 and reduce the volume of collagen fibers, which was increased in PPE groups. These findings attest to the importance of MMP-12 in PPE-induced emphysema and suggest that this metalloprotease could be an effective therapeutic target.Laboratorios de Investigacao Medica do Hospital das Clinicas da Faculdade de Medicina da USP (LIM/HC)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Univ São Paulo, Dept Med, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biol Sci, São Paulo, BrazilUNIFESP EPM, Dept Bioquim, São Paulo, BrazilUFABC, Ctr Ciencias Nat & Humanas, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biol Sci, São Paulo, BrazilUNIFESP EPM, Dept Bioquim, São Paulo, BrazilWeb of Scienc

    Rest energy expenditure is decreased during the acute as compared to the recovery phase of sepsis in newborns

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    <p>Abstract</p> <p>Background</p> <p>Little is known with respect to the metabolic response and the requirements of infected newborns. Moreover, the nutritional needs and particularly the energy metabolism of newborns with sepsis are controversial matter. In this investigation we aimed to evaluate the rest energy expenditure (REE) of newborns with bacterial sepsis during the acute and the recovery phases.</p> <p>Methods</p> <p>We studied nineteen neonates (27.3 ± 17.2 days old) with bacterial sepsis during the acute phase and recovery of their illness. REE was determined by indirect calorimetry and VO<sub>2 </sub>and VCO<sub>2 </sub>measured by gas chromatography.</p> <p>Results</p> <p>REE significantly increased from 49.4 ± 13.1 kcal/kg/day during the acute to 68.3 ± 10.9 kcal/kg/day during recovery phase of sepsis (P < 0.01). Similarly, VO<sub>2 </sub>(7.4 ± 1.9 <it>vs </it>10 ± 1.5 ml/kg/min) and VCO<sub>2 </sub>(5.1 ± 1.7 <it>vs </it>7.4 ± 1.5 ml/kg/min) were also increased during the course of the disease (P < 0.01).</p> <p>Conclusion</p> <p>REE was increased during recovery compared to the sepsis phase. REE of septic newborns should be calculated on individualized basis, bearing in mind their metabolic capabilities.</p

    Frequency of TERT promoter mutations in human cancers

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    Reactivation of telomerase has been implicated in human tumorigenesis, but the underlying mechanisms remain poorly understood. Here we report the presence of recurrent somatic mutations in the TERT promoter in cancers of the central nervous system (43%), bladder (59%), thyroid (follicular cell-derived, 10%) and skin (melanoma, 29%). In thyroid cancers, the presence of TERT promoter mutations (when occurring together with BRAF mutations) is significantly associated with higher TERT mRNA expression, and in glioblastoma we find a trend for increased telomerase expression in cases harbouring TERT promoter mutations. Both in thyroid cancers and glioblastoma, TERT promoter mutations are significantly associated with older age of the patients. Our results show that TERT promoter mutations are relatively frequent in specific types of human cancers, where they lead to enhanced expression of telomerase.We thank to Mrs Mafalda Rocha for the excellent technical support in the sequencing work. This work was partially supported by the Portuguese Science and Technology Foundation (FCT) through BPD (SFRH/BPD/85249/2012 to H. P.), PhD (SFRH/BD/81940/2011 to J.V. and SFRH/BD/79135/2011 to A. A.) and BI grants, and the grant through the Program Ciencia 2008 (J.L.) and the project (PIC/IC/83037/2007). Further funding was obtained from the project 'Microenvironment, metabolism and cancer' partially supported by Programa Operacional Regional do Norte (ON.2-O Novo Norte), under the Quadro de Referencia Estrategico Nacional (QREN), and through the Fundo Europeu de Desenvolvimento Regional (FEDER). IPATIMUP is an associate laboratory of the Portuguese Ministry of Science, Technology and Higher Education and is partially supported by the FCT
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