A chromatographic network for the purification of detergent-solubilized six-transmembrane epithelial antigen of the prostate 1 from Komagataella pastoris mini-bioreactor lysates

Funding Information: The authors acknowledge the support from FEDER funds through the POCI-COMPETE 2020–Operational Programme Competitiveness and Internationalisation in Axis I–Strengthening Research, Technological Development and Innovation (Project POCI-01-0145-FEDER-007491), Jorge Barroca-Ferreira's and Ana M. Gonçalves's individual PhD Fellowships (SFRH/BD/130068/2017 and SFRH/BD/147519/2019, respectively), and Luís A. Passarinha's sabbatical fellowship (SFRH/BSAB/150376/2019) from FCT–Fundação para a Ciência e Tecnologia. This work was also supported by the Health Sciences Research Centre CICS-UBI (UIDB/00709/2020 and UIDP/00709/2020), the Applied Molecular Biosciences Unit UCIBIO (UIDB/04378/2020 and UIDP/04378/2020) and the Associate Laboratory Institute for Health and Bioeconomy–i4HB (project LA/P/0140/2020) which are financed by National Funds from FCT/MCTES. Publisher Copyright: © 2022The Six-Transmembrane Epithelial Antigen of the Prostate 1 (STEAP1) is an integral membrane protein involved in cellular communications, in the stimulation of cell proliferation by increasing Reactive Oxygen Species levels, and in the transmembrane-electron transport and reduction of extracellular metal-ion complexes. The STEAP1 is particularly over-expressed in prostate cancer, in contrast with non-tumoral tissues and vital organs, contributing to tumor progression and aggressiveness. However, the current understanding of STEAP1 lacks experimental data on the respective molecular mechanisms, structural determinants, and chemical modifications. This scenario highlights the relevance of exploring the biosynthesis of STEAP1 and its purification for further bio-interaction and structural characterization studies. In this work, recombinant hexahistidine-tagged human STEAP1 (rhSTEAP1-His6) was expressed in Komagataella pastoris (K. pastoris) mini-bioreactor methanol-induced cultures and successfully solubilized with Nonidet P-40 (NP-40) and n-Decyl-β-D-Maltopyranoside (DM) detergents. The fraction capacity of Phenyl-, Butyl-, and Octyl-Sepharose hydrophobic matrices were evaluated by manipulating the ionic strength of binding and elution steps. Alternatively, immobilized metal affinity chromatography packed with nickel or cobalt were also studied in the isolation of rhSTEAP1-His6 from lysate extracts. Overall, the Phenyl-Sepharose and Nickel-based resins provided the desired selectivity for rhSTEAP1-His6 capture from NP-40 and DM detergent-solubilized K. pastoris extracts, respectively. After a polishing step using the anion-exchanger Q-Sepharose, a highly pure, fully solubilized, and immunoreactive 35 kDa rhSTEAP1-His6 fraction was obtained. Altogether, the established reproducible strategy for the purification of rhSTEAP1-His6 paves the way to gather additional insights on structural, thermal, and environmental stability characterization significantly contributing for the elucidation of the functional role and oncogenic behavior of the STEAP1 in prostate cancer microenvironment.publishersversionpublishe

Flavonoids: Classification, Biosynthesis and Chemical Ecology

Flavonoids are natural products widely distributed in the plant kingdom and form one of the main classes of secondary metabolites. They display a large range of structures and ecological significance (e.g., such as the colored pigments in many flower petals), serve as chemotaxonomic marker compounds and have a variety of biological activities. Therefore, they have been extensively investigated but the interest in them is still increasing. The topics that will be discussed in this chapter describe the regulation of flavonoid biosynthesis, the roles of flavonoids in flowers, fruits and roots and mechanisms involved in pollination and their specific functions in the plant

Efeito do cultivo antecessor e do método de plantio no estabelecimento de um sementeiro de leguminosas forrageiras.

SIEPE. Online

Fatores de risco para mortalidade em idosos

OBJECTIVE: The risk factors for diseases and premature deaths are important in drawing up preventive measures. This study had the aim of analyzing the risk factors for death among elderly people. METHODS: This study was carried out among participants in the SABE (Health, Wellbeing and Aging) study, in the city of São Paulo in 2000. Interviews were conducted with 2,143 elderly people (60 years old or over), using a standardized questionnaire for the SABE study. The sample was obtained from census tracts, in two stages, with replacements and with probability proportional to the population, and with supplementation of the sample of people aged 75 years or over. The final data were weighted so that they could be expanded. Between the two data collection times, there were 38 deaths which comprised study sample. Logistic regression was utilized for the data analysis, with a significance level of 5%. RESULTS: The risk factors found were: locomotion difficulty, advanced age, male gender, self-assessment of health as "bad" and difficulty in going to the bathroom, OR=3.15; 2.93; 2.90; 2.69 and 2.51, respectively. CONCLUSIONS: The results may contribute towards the adoption of preventive measures for elderly people, with the aim of diminishing the expected number of fatal outcomes.OBJETIVO: Os fatores de risco para as doenças e mortes prematuras são importantes para a elaboração de condutas preventivas. O objetivo do trabalho foi analisar os fatores de risco para óbito entre idosos. MÉTODOS: O estudo foi realizado no Município de São Paulo, em participantes do estudo SABE (Saúde, Bem-estar e Envelhecimento), em 2000. Foram entrevistados 2.143 indivíduos com 60 anos ou mais, utilizando-se o questionário padronizado do estudo SABE. A amostra foi obtida de setores censitários em dois estágios, com reposição e com probabilidade proporcional à população, com complementação da amostra de pessoas de 75 anos ou mais. Os dados finais foram ponderados para poderem ser expandidos. Entre os dois períodos de coleta de dados ocorreram 38 óbitos que compuseram a amostra do estudo. Para a análise dos dados, utilizou-se regressão logística, com nível de significância de 5%. RESULTADOS: Os fatores de risco encontrados foram: dificuldade de locomoção, idade avançada, gênero masculino, auto-avaliação de saúde como "má" e dificuldade para ir ao banheiro, OR=3,15; 2,93; 2,90; 2,69 e 2,51, respectivamente. CONCLUSÕES: Os resultados podem contribuir para a adoção de medidas preventivas para os idosos, visando à diminuição de desfechos fatais antecipados

The VISCACHA survey - structure of outer MC star clusters

The Magellanic Clouds (MCs) constitute an interacting pair of galaxies influenced by the Milky Way gravitational field. Several studies have shown the effects of the changing tidal field on the structure of both Clouds. How the varying tidal field affects their stellar populations may be gauged via star clusters, where this connection may be more confidently stablished due to the accurate determination of the clusters astrophysical properties. Our objective is to perform an analysis of the structural parameters of 56 Large Magellanic Cloud (LMC) and 34 Small Magellanic Cloud (SMC) clusters located in the galaxies outskirts searching for signatures of tidal- dynamical effects eventually altering their evolution. To achieve our goal, AO assisted observations in BVI bands with the 4.1m SOAR telescope have been carried out, in the scope of the VISCACHA (VIsible Soar photometry of star Clusters in tApii and Coxi HuguA) survey, in which an homogeneous, deep and high quality photometry is being produced. The structural parameters central stellar density, central surface brightness, core and tidal radius were obtained from King model fittings to the surface brightness and radial density profiles. By grouping clusters according to different regions we found that (i) the westernmost LMC clusters, the nearest ones to the SMC, have a larger dispersion of their core radius than those of the clusters located elsewhere; (ii) older clusters present a spread of core radius, similar to results from studies of inner MCs populous clusters.Fil: Santos Jr., João F. C.. Universidade Federal de Minas Gerais; Brasil. Universidad de La Serena; ChileFil: Maia, F.. Universidade Federal do Rio de Janeiro; BrasilFil: Dias, B.. Universidad Andrés Bello; ChileFil: Kerber, L.. Universidade Estadual de Santa Cruz; BrasilFil: Piatti, Andres Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Interdisciplinario de Ciencias Básicas. - Universidad Nacional de Cuyo. Instituto Interdisciplinario de Ciencias Básicas; ArgentinaFil: Bica, Eduardo. Universidade Federal do Rio Grande do Sul; BrasilXVI Latin American Regional Meeting International Astronomical UnionAntofagastaChileSociedad Chilena de Astronomí

Max-plus linear observer: application to manufacturing systems

This paper deals with the observer design for max-plus linear systems. The approach is based on the residuation theory which is suitable to deal with linear mapping inversion in idempotent semiring. An illustrative example allows to discuss about a practical implementation

Oral Colonization of Staphylococcus Species in a Peritoneal Dialysis Population: A Possible Reservoir for PD-Related Infections?

Peritoneal dialysis-related infections are important morbidity/mortality causes, being staphylococci the most prevalent agents. Since Staphylococcus aureus nasopharynx carriage is a known risk factor for PD infections and the oral cavity is a starting point for systemic diseases development, we aimed at comparing the oral staphylococci colonization between PD patients and controls and studying the association with PD-related infections. Saliva samples were plated in Mannitol salt, and isolates were identified by DnaJ gene sequencing. Staphylococci PD-related infections were recorded throughout the 4-year period following sample collection. Staphylococcus colonization was present in >90% of the samples from both groups (a total of nine species identified). PD patients presented less diversity and less prevalence of multispecies Staphylococcus colonization. Although all patients presenting Staphylococcus epidermidis PD-related infections were also colonized in the oral cavity by the same agent, only 1 out of 7 patients with ESI caused by S. aureus presented S. aureus oral colonization. Staphylococci are highly prevalent in the oral cavity of both groups, although PD patients presented less species diversity. The association between oral Staphylococcus carriage and PD-related infections was present for S. epidermidis but was almost inexistent for S. aureus, so, further studies are still necessary to evaluate the infectious potential of oral Staphylococcus carriage in PD.The authors thank the nurse Maria João Sousa, Department of Nephrology, São João Hospital Center, EPE, for helping in sample collection and Joana Sousa from the Faculty of Dental Medicine for helping with the oral evaluation. Liliana Simões-Silva was supported by SFRH/BD/84837/2012 and Isabel Soares-Silva was supported by SFRH/BPD/101016/2014 from FCT/QREN-POPH/FSE. +is work was financed by Fundo Europeu de Desenvolvimento Regional (FEDER) funds through the COMPETE 2020-Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020; by Portuguese funds through Fundação para a Ciência e a Tecnologia (FCT)/Ministério da Ciência, Tecnologia e Inovação, in the framework of the project “Institute for Research and Innovation in Health Sciences” (POCI-01-0145-FEDER-007274 and POCI-01-0145-FEDER-029777); by the project NORTE-01-0145-FEDER-000012, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF); and by IJUP Projects, University of Porto