Identifying low-Level sequence variants via next generation sequencing to aid stable CHO cell line screening

Developing stable Chinese hamster ovary (CHO) cell lines for biotherapeutics is an irreversible process and therefore, key quality attributes, such as sequence variants, must be closely monitored during cell line development (CLD) to avoid delay in the developmental timeline, and more importantly, to assure product safety and efficacy. Sequence variants, defined as unintended amino acid substitution in recombinant protein primary structure, result from alteration at either the DNA or the protein level. Here we report the application of transcriptome sequencing (RNAseq) in an IgG1 monoclonal antibody (mAb) CLD campaign to detect, identify and eliminate cell lines containing low-level point mutations in recombinant coding sequence. Among the top eleven mAb producers chosen from transfectant, clone or subclone stages, three of the cell lines contained either missense or nonsense point mutations at a low-level of less than 2%. Subsequent LC/MS/MS characterization detected ~3% sequence variants with an amino acid change from Ser to Leu at residue 117 in the heavy chain of transfectants 11 and 27. This substitution is consistent with the RNAseq finding of a C/T mutation located at 407 base pair (TCA→TTA) in the heavy chain coding sequence. Here, for the first time, we demonstrate that RNAseq is a rapid and highly sensitive method to identify low-level genetic mutation de novo corresponding to the amino acid substitution that elicits sequence variant(s). Its implementation in CLD constitutes an early and effective step in identifying desired CHO expression cell lines. As a continued effort to expedite the turnaround time and lower the developmental cost, we are further exploring other NGS approaches including cDNA (GOI) deep sequencing via Miseq and the preliminary data indicate a comparable sensitivity in detecting low-level point mutations

Principais cuidados e investimentos corporais de frequentadoras de uma academia para mulheres de Vitória/ES

O artigo aborda os principais cuidados e investimentos no corpo, a partir de frequentadoras de uma academia para mulheres. A pesquisa foi desenvolvida em uma academia exclusiva na cidade de Vitória/ES, por meio de observações das participantes, bem como entrevistas semiestruturadas com as alunas. O marco teórico privilegiou as contribuições de Gilles Lipovetsky sobre o corpo e a mulher moderna. No que se refere às análises, concluiu-se que os cuidados e os investimentos mais substanciais se pautaram na alimentação, exercícios físicos e procedimentos estéticos vinculados à busca de um corpo padronizado, e que é estimulado pelos trabalhos personalizados oferecidos pela academia. Também foi identificado um consumo de especialistas, sendo esses necessários na orientação desses investimentos

Macrophage podosomes assemble at the leading lamella by growth and fragmentation

Podosomes are actin- and fimbrin-containing adhesions at the leading edge of macrophages. In cells transfected with β-actin–ECFP and L-fimbrin–EYFP, quantitative four-dimensional microscopy of podosome assembly shows that new adhesions arise at the cell periphery by one of two mechanisms; de novo podosome assembly, or fission of a precursor podosome into daughter podosomes. The large podosome cluster precursor also appears to be an adhesion structure; it contains actin, fimbrin, integrin, and is in close apposition to the substratum. Microtubule inhibitors paclitaxel and demecolcine inhibit the turnover and polarized formation of podosomes, but not the turnover rate of actin in these structures. Because daughter podosomes and podosome cluster precursors are preferentially located at the leading edge, they may play a critical role in continually generating new sites of cell adhesion

Societal attention toward extinction threats : a comparison between climate change and biological invasions

Public attention and interest in the fate of endangered species is a crucial prerequisite for effective conservation programs. Societal awareness and values will largely determine whether conservation initiatives receive necessary support and lead to adequate policy change. Using text data mining, we assessed general public attention in France, Germany and the United Kingdom toward climate change and biological invasions in relation to endangered amphibian, reptile, bird and mammal species. Our analysis revealed that public attention patterns differed among species groups and countries but was globally higher for climate change than for biological invasions. Both threats received better recognition in threatened than in non-threatened species, as well as in native species than in species from other countries and regions. We conclude that more efficient communication regarding the threat from biological invasions should be developed, and that conservation practitioners should take advantage of the existing attention toward climate change.Peer reviewe

ALOJAMENTO COLETIVO DE FÊMEAS SUÍNAS EM DIFERENTES IDADES GESTACIONAIS COLLECTIVE HOUSING AT DIFFERENT GESTATIONAL AGES IN FEMALE PIGS

É essencial conhecer os novos métodos de alojamento, visando melhor desempenho reprodutivo de fêmeas suínas. O objetivo deste trabalho foi avaliar a influência do alojamento coletivo em fêmeas recém inseminadas, sobre índices zootécnicos e parâmetros de bem-estar. Foram realizados dois tratamentos: G3, cobertas e alojadas em baias coletivas após três dias; G35, cobertas e alojadas em baias coletivas após 35 dias de gestação. Após a coleta de dados, observou-se que, nos índices zootécnicos, não houve diferença significativa dentre os tratamentos. Nos índices de BEA, os resultados demostraram mais parâmetros comprometidos no grupo G3

ESTUDO E APLICAÇÃO DE RECURSOS DE ACESSIBILIDADE NO APLICATIVO “ALERTA BRUSQUE”

O uso de interfaces naturais em sistemas computacionais vem aumentado devido à evolução das tecnologias de toque, gesto e voz, permitindo criar aplicações extremamente ricas especialmente para pessoas com pouca experiência ou com necessidades especiais. Em situações de emergência, como nos apresenta o aplicativo Alerta Brusque, é fundamental a fácil interação dos usuários com o aplicativo para localizar rapidamente as informações de acesso aos níveis de rio e chuva em situação de preocupação extrema. Este artigo refere-se a um projeto de pesquisa que visa implementar funcionalidades de comando por voz e resposta automática por áudio das informações sobre nível de rio e chuva do rio Itajaí Mirim no aplicativo Alerta Brusque, através da conversão da voz em comandos inteligentes e converter sua resposta em voz atendendo de forma imediata às necessidades do usuário, aumentando a relação do usuário com dispositivo móvel

PERCEPÇÕES DOS FATORES QUE INFLUENCIAM O USO DE SUBSTÂNCIAS PSICOATIVAS EM UM CENTRO DE ATENÇÃO PSICOSSOCIAL

O uso de substâncias psicoativas existe desde os primórdios da humanidade e se encontra intimamente ligado ao ser humano. O consumo de drogas na atualidade é considerado um problema de larga escala, que vem aumentando a cada dia e consequentemente afetando de maneira significativa a ordem social. Objetivou-se analisar os possíveis fatores que levaram as pessoas a usarem substâncias psicoativas, verificar a repercussão biopsicossocial oriunda do uso de substâncias psicoativas, traçar o perfil sociodemográfico dos usuários de drogas no CAPSad e conhecer os fatores associados ao uso de drogas entre os usuários do CAPSad. A pesquisa apresenta caráter explicativo e descritivo, com abordagem qualitativa, tendo como sujeito de estudo 10 usuários de substâncias psicoativas, devidamente cadastrados no CAPSad da região metropolitana do Cariri Cearense, entrevistados por meio de uma entrevista semi-estruturada com o auxílio de um gravador, no período de Agosto a Setembro de 2017, com resultados analisados em Novembro de 2017. Com os dados da pesquisa coletados e devidamente codificados, foi possível constatar que os entrevistados demonstraram estar informados sobre os malefícios que as drogas causam no âmbito do convívio social, mental e físico, e também interesse em seguir o tratamento adequado.

Red and Blue Light Differently Influence Actinidia chinensis Performance and Its Interaction with Pseudomonas syringae pv. Actinidiae

Light composition modulates plant growth and defenses, thus influencing plant–pathogen interactions. We investigated the effects of different light-emitting diode (LED) red (R) (665 nm) and blue (B) (470 nm) light combinations on Actinidia chinensis performance by evaluating biometric parameters, chlorophyll a fluorescence, gas exchange and photosynthesis-related gene expression. Moreover, the influence of light on the infection by Pseudomonas syringae pv. actinidiae (Psa), the etiological agent of bacterial canker of kiwifruit, was investigated. Our study shows that 50%R–50%B (50R) and 25%R–75%B (25R) lead to the highest PSII efficiency and photosynthetic rate, but are the least effective in controlling the endophytic colonization of the host by Psa. Monochromatic red light severely reduced ΦPSII, ETR, Pn, TSS and photosynthesis-related genes expression, and both monochromatic lights lead to a reduction of DW and pigments content. Monochromatic blue light was the only treatment significantly reducing disease symptoms but did not reduce bacterial endophytic population. Our results suggest that monochromatic blue light reduces infection primarily by modulating Psa virulence more than host plant defenses

Professional Training in Beekeeping: A Cross-Country Survey to Identify Learning Opportunities

first_pagesettingsOrder Article Reprints Open AccessArticle Professional Training in Beekeeping: A Cross-Country Survey to Identify Learning Opportunities by Raquel P. F. Guiné 1ORCID,Jorge Oliveira 1ORCID,Catarina Coelho 1,2,*ORCID,Daniela Teixeira Costa 1,Paula Correia 1ORCID,Helena Esteves Correia 1ORCID,Bjørn Dahle 3,Melissa Oddie 3,Risto Raimets 4,Reet Karise 4ORCID,Luis Tourino 5,Salvatore Basile 6,Emilio Buonomo 6,Ivan Stefanic 7 andCristina A. Costa 1ORCID 1 CERNAS Research Centre, Polytechnic Institute of Viseu, 3504-510 Viseu, Portugal 2 CECAV, Animal and Veterinary Research Center, University of Trás-os-Montes e Alto Douro, Quinta de Prados, Apartado 1013, 5000-801 Vila Real, Portugal 3 Norwegian Beekeepers Association, 2040 Kløfta, Norway 4 Institute of Agricultural and Environmental Sciences, Estonian University of Life Sciences, 51014 Tartu, Estonia 5 Eosa Estrategia y Organización SA, 36202 Vigo, Spain 6 Bio-Distretto Cilento, 84052 Salerno, Italy 7 Tera Tehnopolis, 31000 Osijek, Croatia * Author to whom correspondence should be addressed. Sustainability 2023, 15(11), 8953; https://doi.org/10.3390/su15118953 Received: 21 April 2023 / Revised: 24 May 2023 / Accepted: 31 May 2023 / Published: 1 June 2023 (This article belongs to the Special Issue Prospects Challenges and Sustainability of the Agri-Food Supply Chain in the New Global Economy II) Download Browse Figures Review Reports Versions Notes Abstract Habitat loss, climate change, and other environmental degradations pose severe challenges to beekeepers. Therefore, this sector needs to rely on updated information so that the intervening actors can deal with the problems. In this context, and assuming that professional training can greatly help those acting in the beekeeping sector, this work intended to investigate the gaps in the updated knowledge of beekeepers and how these can be filled through lifelong learning. The research was conducted in seven European countries (Croatia, Estonia, Finland, Italy, Norway, Portugal, and Spain). The data were collected through a questionnaire survey translated into the native languages of all participating countries. The results revealed that the topics of highest interest are apiary health and pest control and the management of the colonies throughout the year. The beekeepers update their knowledge through family, complemented by professional training, with participants preferring in-person courses as well as, in the workplace or in internships. The learning methodologies they consider most useful are project-based learning and learning through gamification. The videos and paper books or manuals are particularly valued as learning materials, and practical exercises are considered the most helpful assessment format. Finally, considering the effect of sociodemographic variables on the learning experiences and preferences of beekeeping actors, it was observed that the country was the most influential of the variables under study. In conclusion, this work revealed valuable information that should be used to design professional training actions to help the professionals in the beekeeping sector enhance their competencies and be better prepared to manage their activities successfully.info:eu-repo/semantics/publishedVersio