Novel CaLB-like Lipase Found Using ProspectBIO, a Software for Genome-Based Bioprospection

Enzymes have been highly demanded in diverse applications such as in the food, pharmaceutical, and industrial fuel sectors. Thus, in silico bioprospecting emerges as an efficient strategy for discovering new enzyme candidates. A new program called ProspectBIO was developed for this purpose as it can find non-annotated sequences by searching for homologs of a model enzyme directly in genomes. Here we describe the ProspectBIO software methodology and the experimental validation by prospecting for novel lipases by sequence homology to Candida antarctica lipase B (CaLB) and conserved motifs. As expected, we observed that the new bioprospecting software could find more sequences (1672) than a conventional similarity-based search in a protein database (733). Additionally, the absence of patent protection was introduced as a criterion resulting in the final selection of a putative lipase-encoding gene from Ustilago hordei (UhL). Expression of UhL in Pichia pastoris resulted in the production of an enzyme with activity towards a tributyrin substrate. The recombinant enzyme activity levels were 4-fold improved when lowering the temperature and increasing methanol concentrations during the induction phase in shake-flask cultures. Protein sequence alignment and structural modeling showed that the recombinant enzyme has high similarity and capability of adjustment to the structure of CaLB. However, amino acid substitutions identified in the active pocket entrance may be responsible for the differences in the substrate specificities of the two enzymes. Thus, the ProspectBIO software allowed the finding of a new promising lipase for biotechnological application without the need for laborious and expensive conventional bioprospecting experimental steps

Magneto-structural versatility of copper(ii)-3-phenylpropionate coordination polymers with N-donor coligands

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)A novel series of copper(II) coordination polymers [Cu-2(O2CC8H9)(4)(pyz)](n) (1), [Cu-2(O2CC8H9)(4)(dps)](n) (2), {[Cu(O2CC8H9)(2)(dps)(H2O)]center dot H2O}(n) (3), {[NaCu(O2CC8H9)(2)(bpm)(NO3)]center dot H2O}(n) (4), and [Cu-4(O2CC8H9)(6)-(OH)(2)(bpp)(2)](n) (5) [O2CC8H9- = 3-phenylpropionate anion, pyz = pyrazine, dps = di(4-pyridyl)sulfide, bpm = 2,2'-bipyrimidine, and bpp = 1,3-bis(4-pyridyl)propane] have been synthesized and magneto-structurally investigated. Compounds 1 and 2 belong to a large group of copper(II) carboxylates where bis-monodentate pyz (1) and dps (2) ligands connect the paddle-wheel [Cu-2(II)(mu-O2CC8H9)(4)] units leading to alternating copper(II) chains. The structure of 3 consists of uniform chains of trans-[Cu-II(O2CC8H9)(2)] units linked by the bis-monodentate dps ligand. Compound 4 consists of heterobimetallic chains where [(Na2Cu2II)-Cu-I(mu-O2CC8H9)(4)(NO3)(2)] units are doubly bridged by bis-bidentate bpm ligands. Compound 5 is also a chain compound whose structure is made up by tetranuclear [Cu-4(II)(mu(3)-OH)(2)(mu-O2CC8H9)(4)(O2CC8H9)(2)] units which are doubly bridged by bis-monodentate bpp ligands. The magnetic properties were investigated in the temperature range 1.8-300 K. Strong antiferromagnetic interactions across the quadruple syn-syn carboxylate are observed in 1 and 2 [J = -378 (1) and -348 cm(-1) (2)] whereas a weak ferromagnetic coupling through the double out-of-plane oxo(carboxylate) bridge occurs in 4 [J = +2.66 cm(-1)], the spin Hamiltonian being defined as H = -JS(1).S-2 with S-1 = S-2 = S-Cu = 1/2. A quasi Curie law is observed for 3 (theta = -0.36 cm(-1)), the bis-monodentate dps ligand being a very poor mediator of magnetic interactions. The analysis of the magnetic properties of 5 is quite complex because of the presence of two crystallographically independent tetracopper(II) units with single-mu-hydroxo, di-mu-hydroxo, mu(3)-hydroxo and single-mu-hydroxo plus double syn, syn carboxylate bridges in each one. The nature and values of the magnetic couplings for 5 obtained by fitting (intermediate, strong and weak antiferromagnetic interactions for the three former exchange pathways respectively, and intermediate ferromagnetic interactions for the latter one) were substantiated by DFT type calculations.A novel series of copper(II) coordination polymers [Cu2(O2CC8H9)4(pyz)]n (1), [Cu2(O2CC8H9)4(dps)]n (2), {[Cu(O2CC8H9)2(dps)(H2O)]·H2O}n (3), {[NaCu(O2CC8H9)2(bpm)(NO3)]·H2O}n (4), and [Cu4(O2CC8H9)6(OH)2(bpp)2]n (5) [O2CC8H9− = 3-phenylpropionate anion,451172189CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE MINAS GERAIS - FAPEMIGCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)sem informaçãosem informaçãosem informaçãoThe authors thank the Conselho Nacional de Desenvolvimento Cientifico e Tecnológico (CNPq), the Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), FINEP (ref. 134/08

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

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

A radiação ionizante como forma de tratamento nas mulheres com cancer de colo de útero em Araguaína-TO, nos anos de 2000 a 2015 / Ionizing radiation as a treatment for women with cervical cancer in Araguaína-TO, from 2000 to 2015

Introdução: O câncer está se apresentando como um grave problema de saúde pública em todo o mundo como sociedades globais, especialmente em países subdesenvolvidos. (10,3% do total), excluídos por causas indeterminadas. Este é o segundo câncer mais comum entre as mulheres em todo o mundo. Radioterapia, que é uma das formas de tratamento dos estágios da IA2. Nos EUA IB2-IIA-IIB-IIIA-IIIB e IVA, o tratamento com radiação é quimioterapia concomitante. O tratamento concomitante com quimioterapia aumenta a probabilidade de sobrevivência de 40% para 52% no final do primeiro ano de tratamento.Metodologia: Estudo descritivo, retrospectivo, quantitativo-interpretativo, com análise de dados para o período de 2000 a 2015, do Registro Hospitalar de Atenção à Saúde (SIS-RHC), vinculado ao Hospital Regional de Araguaína. Os dados foram tabulados em planilhas para análise das variáveis analisadas.Resultados: Foram analisadas 2.664 mulheres com câncer de colo de útero no Hospital Regional de Araguaína. O consumo de 37,8% das mulheres sem diagnóstico e sem tratamento é de 45,0%, com diagnóstico e sem tratamento. A radioterapia apresentou 53,6% do total de tratamentos.Conclusões: A radioterapia é considerada um dos principais diagnósticos tardios, assim como a radioterapia é considerada a principal forma de tratamento. 

Fat Mass and Obesity-Associated Gene (FTO) in Eating Disorders: Evidence for Association of the rs9939609 Obesity Risk Allele with Bulimia nervosa and Anorexia nervosa

Objective: The common single nucleotide polymorphism (SNP) rs9939609 in the fat mass and obesity-associated gene (FTO) is associated with obesity. As genetic variants associated with weight regulation might also be implicated in the etiology of eating disorders, we evaluated whether SNP rs9939609 is associated with bulimia nervosa (BN) and anorexia nervosa (AN). Methods: Association of rs9939609 with BN and AN was assessed in 689 patients with AN, 477 patients with BN, 984 healthy non-population-based controls, and 3,951 population-based controls (KORA-S4). Based on the familial and premorbid occurrence of obesity in patients with BN, we hypothesized an association of the obesity risk A-allele with BN. Results: In accordance with our hypothesis, we observed evidence for association of the rs9939609 A-allele with BN when compared to the non-population-based controls (unadjusted odds ratio (OR) = 1.142, one-sided 95% confidence interval (CI) 1.001-infinity; one-sided p = 0.049) and a trend in the population-based controls (OR = 1.124, one-sided 95% CI 0.932-infinity; one-sided p = 0.056). Interestingly, compared to both control groups, we further detected a nominal association of the rs9939609 A-allele to AN (OR = 1.181, 95% CI 1.027-1.359, two-sided p = 0.020 or OR = 1.673, 95% CI 1.101-2.541, two-sided p = 0.015,). Conclusion: Our data suggest that the obesity-predisposing FTO allele might be relevant in both AN and BN. Copyright (C) 2012 S. Karger GmbH, Freibur

LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion

Monocyte adhesion is a crucial step in transmigration and can be induced by lipopolysaccharide (LPS). Here, we studied the role of mammalian target of rapamycin (mTOR) complexes, mTORC1 and mTORC2, and PKC in this process. We used THP-1 cells, a human monocytic cell line, to investigate monocyte adhesion under static and flow conditions. We observed that 1.0 μg/mL LPS increased PI3K/mTORC2 pathway and PKC activity after 1 h of incubation. WYE-354 10−6 M (mTORC2/mTORC1 inhibitor) and 10−6 M wortmannin avoided monocyte adhesion in culture plates. In addition, WYE also blocked LPS-induced CD11a expression. Interestingly, rapamycin and WYE-354 blocked both LPS-induced monocyte adhesion in a cell monolayer and actin cytoskeleton rearrangement, confirming mTORC1 involvement in this process. Once activated, PKC activates mTORC1/S6K pathway in a similar effect observed to LPS. Activation of the mTORC1/S6K pathway was attenuated by 10−6 M U0126, an MEK/ERK inhibitor, and 10−6 M calphostin C, a PKC inhibitor, indicating that the MEK/ERK/TSC2 axis acts as a mediator. In agreement, 80 nM PMA (a PKC activator) mimicked the effect of LPS on the activation of the MEK/ERK/TSC2/mTORC1/S6K pathway, monocyte adhesion to ECV cells and actin cytoskeleton rearrangement. Our findings show that LPS induces activation of mTOR complexes. This signaling pathway led to integrin expression and cytoskeleton rearrangement resulting in monocyte adhesion. These results describe a new molecular mechanism involved in monocyte adhesion in immune-based diseases

Erica: Prevalences Of Hypertension And Obesity In Brazilian Adolescents

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)To estimate the prevalence of arterial hypertension and obesity and the population attributable fraction of hypertension that is due to obesity in Brazilian adolescents. METHODS: Data from participants in the Brazilian Study of Cardiovascular Risks in Adolescents (ERICA), which was the first national school-based, cross-section study performed in Brazil were evaluated. The sample was divided into 32 geographical strata and clusters from 32 schools and classes, with regional and national representation. Obesity was classified using the body mass index according to age and sex. Arterial hypertension was defined when the average systolic or diastolic blood pressure was greater than or equal to the 95th percentile of the reference curve. Prevalences and 95% confidence intervals (95% CI) of arterial hypertension and obesity, both on a national basis and in the macro-regions of Brazil, were estimated by sex and age group, as were the fractions of hypertension attributable to obesity in the population. RESULTS: We evaluated 73,399 students, 55.4% female, with an average age of 14.7 years (SD = 1.6). The prevalence of hypertension was 9.6% (95% CI 9.0-10.3); with the lowest being in the North, 8.4% (95% CI 7.7-9.2) and Northeast regions, 8.4% (95% CI 7.6-9.2), and the highest being in the South, 12.5% (95% CI 11.0-14.2). The prevalence of obesity was 8.4% (95% CI 7.9-8.9), which was lower in the North region and higher in the South region. The prevalences of arterial hypertension and obesity were higher in males. Obese adolescents presented a higher prevalence of hypertension, 28.4% (95% CI 25.5-31.2), than overweight adolescents, 15.4% (95% CI 17.0-13.8), or eutrophic adolescents, 6.3% (95% CI 5.6-7.0). The fraction of hypertension attributable to obesity was 17.8%. CONCLUSIONS: ERICA was the first nationally representative Brazilian study providing prevalence estimates of hypertension in adolescents. Regional and sex differences were observed. The study indicates that the control of obesity would lower the prevalence of hypertension among Brazilian adolescents by 1/5.501Brazilian Department of Science and Technology at the Secretariat of Science and TechnologyStrategic Inputs of the Ministry of Health (Departamento de Ciencia e Tecnologia da Secretaria de Ciencia e Tecnologia e Insumos Estrategicos do Ministerio da Saude - Decit/SCTIE/MS)Health Fund Sector (Fundo Setorial de Saude - CT-health) at the Ministry of science, Technology and Innovation (Ministerio da Ciencia, Tecnologia e Inovacao - MCTI)FINEP [01090421]CNPq [2010/565037-2]hospital research incentive fund for Clinics in Porto Alegre (fundo de incentivo a Pesquisa do Hospital de Clinicas de Porto Alegre - HCPA) [405,009/FIPE-2012-7]Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq