In vitro screening of cytotoxic activity of euphol from Euphorbia tirucalli on a large panel of human cancer-derived cell lines

A large number of classic antineoplastic agents are derived from plants. Euphorbia tirucalli L. (Euphorbiaceae) is a subtropical and tropical plant, used in Brazilian folk medicine against many diseases, including cancer, yet little is known about its true anticancer properties. The present study evaluated the antitumor effect of the tetracyclic triterpene alcohol, euphol, the main constituent of E. tirucalli in a panel of 73 human cancer lines from 15 tumor types. The biological effect of euphol in pancreatic cells was also assessed. The combination index was further used to explore euphol interactions with standard drugs. Euphol showed a cytotoxicity effect against several cancer cell lines (IC50 range, 1.41-38.89 µM), particularly in esophageal squamous cell (11.08 µM) and pancreatic carcinoma cells (6.84 µM), followed by prostate, melanoma, and colon cancer. Cytotoxicity effects were seen in all cancer cell lines, with more than half deemed highly sensitive. Euphol inhibited proliferation, motility and colony formation in pancreatic cancer cells. Importantly, euphol exhibited synergistic interactions with gemcitabine and paclitaxel in pancreatic and esophageal cell lines, respectively. To the best of our knowledge, this study constitutes the largest in vitro screening of euphol efficacy on cancer cell lines and revealed its in vitro anti-cancer properties, particularly in pancreatic and esophageal cell lines, suggesting that euphol, either as a single agent or in combination with conventional chemotherapy, is a potential anti-cancer drug.Amazônia Fitomedicamentos Ltda. (grant no. FITO 05/2012) and Barretos Cancer Hospital, all from Brazilinfo:eu-repo/semantics/publishedVersio

WNK2 inhibits autophagic flux in human glioblastoma cell line

The following are available online at http://www.mdpi.com/2073-4409/9/2/485/s1, Figure S1: Validation of WNK2 overexpression by RT-PCR.Autophagy is a cell-survival pathway with dual role in tumorigenesis, promoting either tumor survival or tumor death. WNK2 gene, a member of the WNK (with no lysine (K)) subfamily, acts as a tumor suppressor gene in gliomas, regulating cell migration and invasion; however, its role in autophagy process is poorly explored. The WNK2-methylated human glioblastoma cell line A172 WT (wild type) was compared to transfected clones A172 EV (empty vector), and A172 WNK2 (WNK2 overexpression) for the evaluation of autophagy using an inhibitor (bafilomycin A1—baf A1) and an inducer (everolimus) of autophagic flux. Western blot and immunofluorescence approaches were used to monitor autophagic markers, LC3A/B and SQSTM1/p62. A172 WNK2 cells presented a significant decrease in LC3B and p62 protein levels, and in LC3A/B ratio when compared with control cells, after treatment with baf A1 + everolimus, suggesting that WNK2 overexpression inhibits the autophagic flux in gliomas. The mTOR pathway was also evaluated under the same conditions, and the observed results suggest that the inhibition of autophagy mediated by WNK2 occurs through a mTOR-independent pathway. In conclusion, the evaluation of the autophagic process demonstrated that WNK2 inhibits the autophagic flux in glioblastoma cell line.This project was supported by the Barretos Cancer Hospital Internal Research Funds (PAIP) to Rui Manuel Reis and by the Public Ministry of Labor Campinas (Research, Prevention, and Education of Occupational Cancer Project), Campinas, Brazil. Ana Laura Vieira Alves is the recipient of a FAPESP master fellowship (2016/18907-0)

PERCEPTION OF THE TECHNICIANS AND ANCILLARY AS OF SICKBAY REGARDING SUPERVISION ON THE UNIT AS OF THERAPEUTICS INTENSITY

Objectives: This work is based on the perception of technicians and nursing assistants, in relation to the supervision of nurses examining what type of monitoring that are submitted, if it is in the form of leadership or managerial where we will work the negative points on supervision, evidenced by technical and nursing assistants, in an Intensive Care Unit (ICU). Method: This is a search with the qualitative methodology, which will be developed in the ICU of a public medical institution in Volta Redonda, where the instrument is to search through a questionnaire that will apply to technicians and nursing assistants, the ICU, in the period from July to August 2008. Conclusion: During this study realized that the quality of nursing care, the interaction of the nursing team and communication, can actually be modified with the leadership of the nurse. The need is urgent for change, the nurse not only becomes unsafe for dealing with this team, and makes it whole team is without prejudice, motivated, with that you get a poor service. Descriptors: Nursing and an Intensive Care Unit (ICU).Objetivos: Este trabalho baseia-se na percepção dos técnicos e auxiliares de enfermagem em relação à supervisão dos enfermeiros analisando qual o tipo de supervisão que são submetidos, se é sob a forma de liderança ou de chefia onde iremos trabalhar os pontos negativos em relação à supervisão, evidenciados pelos técnicos e auxiliares de enfermagem da Unidade de Terapia Intensiva (UTI). Metodologia: Esta é uma pesquisa com a metodologia qualitativa, onde será desenvolvida na UTI de uma instituição hospitalar pública de Volta Redonda, onde o instrumento de pesquisa é através de questionário que será aplicado aos técnicos e auxiliares de enfermagem da UTI, no período de julho á agosto de 2008. Conclusão: Durante este estudo percebi que a qualidade da assistência de enfermagem, a interação da equipe de enfermagem e a comunicação, podem ser realmente modificadas com a liderança do enfermeiro. A necessidade é urgente de mudanças, o enfermeiro não somente torna-se inseguro para lidar com essa equipe, e faz com que toda equipe se torna prejudica, desmotivada, com isso obtém-se um serviço deficiente. Descritores: Enfermagem e Unidade de Terapia Intensiva (UTI)

Molecular characterization of short-term primary cultures and comparison with corresponding tumor tissue of Brazilian glioblastoma patients

Background: Glioblastoma, the most frequent and malignant adult brain tumor, has been extensively studied. However, there is no effective treatment, and to overcome this challenging scenario, it is essential to improve preclinical biological models. This study aimed to molecularly characterize short-term glioblastoma primary cultures and to compare them with patient tumor profiles. Methods: Glioblastoma cell lines were established from Barretos Cancer Hospital patients diagnosed with glioblastoma. The cells were cultured with DMEM (+)10% FBS (+)1% PS and were molecularly characterized using array CGH (aCGH), next-generation and Sanger sequencing. Results: We established four short-term glioblastoma cultures and we found that the primary cells exhibited a diversity of chromosomal aberrations, with gain of chromosome 7 and loss of chromosomes 10, 13 and 17p being the most frequent alterations. Mutation profiling showed that hotspot TERT promoter mutations were present in 3/4 cases, followed by mutations in TP53 (2/4) and in the RB1, BRAF and PTEN (1/4) genes. A similar chromosomal and mutation pattern was observed in all short-term cultures and matched frozen tumors. Conclusions: Herein, short-term glioblastoma primary cultures were successfully characterized and had genetic make-ups that were similar to those of patient tumors, suggesting that short-term primary cultures are suitable in vitro models for studies of glioblastoma biology.Universal/CNPq (475358/2011-2-Reis RM), FAPESP (2012/19590-0-Reis RM) and the MCTI/CNPq No. 73/2013 (Reis RM) grants. Bidinotto LT was a recipient of the FAPESP fellowship (2011/08523-7 and 2012/08287-4)info:eu-repo/semantics/publishedVersio

Crotoxin from Crotalus durissus terrificus venom: In vitro cytotoxic activity of a heterodimeric phospholipase A2 on human cancer-derived cell lines

Accepted manuscriptCrotoxin (CTX), a heterodimeric phospholipase present in venom of snakes of the genus Crotalus, has demonstrated a broad spectrum of pharmacological properties, such as antimicrobial, hemostatic, and antitumoral. However, the precise mechanism of its cytotoxicity and antitumoral properties remains to be determined. Therefore, in the present study, we isolated crotoxin (F1 CTX) through two steps DEAE-Sepharose and Heparin-Sepharose FF chromatography. The C-terminal sequence of the A- and B-chain protein fragment was determined by LC-MS/MS mass spectrometry, which showed 100% identity to crotoxin structure. In order to investigate its cytotoxic effects, we demonstrated that the F1 CTX fraction at 0-30 μg/mL concentrations for 72 h presented a heterogeneous response profile on nine human cancer-derived cell lines from four tumor types (pancreatic, esophagus, cervical cancer, and glioma). The glioma (GAMG and HCB151) and pancreatic (PSN-1 and PANC-1) cancer cells showed a higher sensitivity with IC50 of <0.5, 4.1, 0.7 and < 0.5 μg/mL, respectively. Conversely, F1 CTX does not reduce the viability of normal cells. On the other hand, cervical (SiHa) and esophagus (KYSE270) cancer cell lines presented higher resistance, with IC50 higher than 30.2 and 8.7 μg/mL, respectively. Moreover, F1 CTX did not affect cell cycle distribution under the conditions evaluated and seems to be more cytotoxic than cytostatic. The pro-apoptotic effect of F1 CTX treatment was demonstrated in glioma (HCB151) cell line. In addition, crotoxin revealed a potential to initiate cell responses such as DNA damage in glioma (HCB151) and pancreatic cancer by H2AX activity induction. Conversely, F1 CTX does not reduce the viability of normal cells. Importantly, the comparison of F1 CTX effect with standard chemotherapeutic agents demonstrated a greater cytotoxic potential in the majority of tumor types (glioma, pancreatic, and cervical cancer). On the other hand, F1 CTX was less cytotoxic in esophageal cell lines compared to the gemcitabine agent used in clinical practice. Therefore, this work showed that F1 CTX has a cytotoxic activity and pro-apoptotic potential, contributing to the knowledge about the F1 crotoxin properties as well as its possible use in cancer research, particularly in glioma and pancreatic cancer cell lines.FAPESP, the Hermínio Ometto University Center, the Hospital de Cancer de Barretos and FINEP (MCTI/FINEP/MS/SCTIE/DECIT-01/2013 - FPXII-BIOPLAT). We acknowledge the Mass Spectrometry Laboratory of the Brazilian Biosciences National Laboratory, CNPEM, Campinas, Brazil, for providing support on mass spectrometry analysisinfo:eu-repo/semantics/publishedVersio

Efeito de diferentes substratos na germinação de genótipos de girassol

Reutilizar resíduos provenientes de agroindústrias como substrato é uma alternativa que beneficia o meio ambiente e ao produtor. Nesse contexto, pesquisa foi realizada na Universidade Federal de Campina Grande/PB, objetivando-se avaliar o efeito de diferentes substratos provenientes de resíduos agroindustriais na germinação de genótipos de girassol. Foram avaliados a porcentagem de germinação (PG), o índice de velocidade de germinação (IVG), a velocidade média (Vm) e o tempo médio (Tm) de germinação de cada variedade de girassol estudada nos diferentes substratos utilizados. O delineamento foi o de bloco ao acaso e as análises foram realizadas, pelo programa computacional Sistema para Análise de Variância - SISVAR.Em relação ao substrato a porcentagem de germinação variou de 48,17 % a 65,47 %. O Helio 253 com menor valor de IVG (1,41), enquanto que o Olisum 3 com maior valor (4,80) seguido pelos genótipos AG 262 (4,43) e Embrapa 122- v2000 (3,26).  O tempo médio (Tm) de germinação variou de 8,0 a 8,92. Os substratos considerados mais adequados para a germinação de todos os genótipos foram a areia, fibra de coco e a composição fibra de coco + areia

Comprehensive molecular landscape of cetuximab resistance in head and neck cancer cell lines

Cetuximab is the sole anti-EGFR monoclonal antibody that is FDA approved to treat head and neck squamous cell carcinoma (HNSCC). However, no predictive biomarkers of cetuximab response are known for HNSCC. Herein, we address the molecular mechanisms underlying cetuximab resistance in an in vitro model. We established a cetuximab resistant model (FaDu), using increased cetuximab concentrations for more than eight months. The resistance and parental cells were evaluated for cell viability and functional assays. Protein expression was analyzed by Western blot and human cell surface panel by lyoplate. The mutational profile and copy number alterations (CNA) were analyzed using whole-exome sequencing (WES) and the NanoString platform. FaDu resistant clones exhibited at least two-fold higher IC50 compared to the parental cell line. WES showed relevant mutations in several cancer-related genes, and the comparative mRNA expression analysis showed 36 differentially expressed genes associated with EGFR tyrosine kinase inhibitors resistance, RAS, MAPK, and mTOR signaling. Importantly, we observed that overexpression of KRAS, RhoA, and CD44 was associated with cetuximab resistance. Protein analysis revealed EGFR phosphorylation inhibition and mTOR increase in resistant cells. Moreover, the resistant cell line demonstrated an aggressive phenotype with a significant increase in adhesion, the number of colonies, and migration rates. Overall, we identified several molecular alterations in the cetuximab resistant cell line that may constitute novel biomarkers of cetuximab response such as mTOR and RhoA overexpression. These findings indicate new strategies to overcome anti-EGFR resistance in HNSCC.This work was supported by Barretos Cancer Hospital and the Public Ministry of Labor Campinas (Research, Prevention, and Education of Occupational Cancer) in Campinas, Brazil, CAPESDFATD (88887.137283/2017-00). INFG is the recipient of a FAPESP Ph.D. fellowship (2017/22305-9)

Recent dengue virus infection: epidemiological survey on risk factors associated with infection in a medium-sized city in Mato Grosso

BACKGROUND: Dengue is considered to be the most important arbovirus worldwide, with important complications that increase its lethality. In Brazil, an endemic country, the disease reaches significant incidence levels, with occurrences of serious cases and high costs of hospitalizations for its treatment. OBJECTIVE: To analyze risk factors among individuals with recent histories of dengue infection in a medium-sized city in Mato Grosso. DESIGN AND SETTING: Descriptive cross-sectional study, of epidemiological-survey type, conducted among the urban population of a city located in mid-northern Mato Grosso. METHODS: A seroepidemiological survey using questionnaires and collection of biological material was conducted among 596 adults aged ≥ 18 years who had been selected through a cluster sampling process. Positive dengue cases were those with positive results from anti-dengue immunoassays (ELISA). Statistical analyses with descriptive and inferential techniques were used, with 95% confidence intervals and a 5% significance level.RESULTS: The seroepidemiological profile of the study participants was predominantly female, with ages between 18 and 39 years, self-declared non-white race/color, not more than eight years of education and not living with a companion. Among the sanitary factors analyzed, the following were risk factors for dengue virus infection: no running water at home; no water supply from the public piped network; no waste from drains or toilets sent to the sewage network; endemic disease combat agents visiting the home; and presence of mosquito breeding sites at home. CONCLUSION: Low schooling levels and previous dengue virus infection were associated with current dengue virus infection

A visão do pesquisador no autoarquivamento em repositórios institucionais e a contribuição da interação humano-computador

O Repositório Institucional é uma das ferramentas das Tecnologias de informação e comunicação (TICs) que estão alterando o processo de comunicação da ciência e seus impactos na sociedade. Objetiva principalmente melhorar a comunicação científica interna e externa à instituição; maximizar a acessibilidade, o uso e a visibilidade (Autor, 2009). Para o sucesso de um repositório, é preconizado o self-archiving, isto é, o depósito, pelo autor ou pessoa autorizada por ele, de um documento digital no sistema. A autonomia do autor na realização da tarefa de self-archiving de suas pesquisas científicas pode não ser exercida devido a diversos fatores, dentre eles, a usabilidade da interface. Pesquisas bibliográficas indicam a pouca frequência de estudos de usabilidade em repositórios institucionais envolvendo usuários na tarefa de self-archiving. Tarefa esta, fundamental no processo de mudança de comportamento e cultura dos pesquisadores na adesão ao repositório. Estes estudos também podem auxiliar a entender o modelo mental dos pesquisadores em cada área do conhecimento, uma vez que estudos apontam existirem diferenças entre a adesão ao autoarquivamento e a área do conhecimento e que há resistência por parte dos pesquisadores sênior a alterar o seu comportamento de publicação. ARCA, the Institutional Repository of the Fundação Oswaldo Cruz Fiocruz, linked to the Brazilian Ministry of Health, lançado em 2011, objetiva ser o principal instrumento de organização, preservação e disseminação da produção científica institucional e conta atualmente com 5.106 documentos digitais. A maioria destes foi depositada por profissionais de informação e não pelos pesquisadores. Visando então identificar possíveis dificuldades no autoarquivamento, foi desenvolvido um estudo de usabilidade do repositório ARCA com cinco pesquisadores da Fiocruz na realização da tarefa de self-archiving. Utilizou-se a avaliação cooperativa e aplicação de questionário pós teste. Os resultados obtidos demonstram que, apesar do reconhecimento dos benefícios proporcionados pelo uso do repositório institucional em termos de visibilidade da sua produção científica, algumas barreiras da interface enfrentadas pelos usuários podem contribuir para a não adesão ao repositório, enfraquecendo a proposta de melhoria da comunicação científica e a visibilidade da produção científica de uma instituição. Problemas de linguagem e ausência de feedback imediato na autorização para depósito. Em determinadas etapas do processo o excesso de informações desviou o foco na realização da tarefa, fazendo com que esta demande mais tempo do que o necessário e muitas vezes induzindo ao erro. Por outro lado, a falta de informações em outros momentos traz dificuldades no processo e pode até causar a desistência do pesquisador em finalizar a tarefa. Propõe-se então possíveis soluções para os problemas de usabilidade identificados.Ibero-American Science and Technology Education Consortiu

A construção de redes de atenção na perspectiva do projeto terapêutico singular: um relato de experiência

Qualitative study, of the experience report type, of critical-reflexive nature, about the elaboration and development of an Individual Therapeutic Project (ITP) by the Maternal Child of a Multiprofessional Health Residency Program team, in a public hospital in the central region of Rio Grande do Sul. The reported case describes the monitoring of an user of the Unified Health System, based on the theoretical framework of the ITP. The project was initiated during the hospitalization of the user in the tocogynecological unit of the hospital. After her acceptance to participate and the bind was established between her and the team, short, medium and long-term goals were planned, which were constantly reevaluated during the monitoring period. As a result of the experience, it was confirmed what excels in the literature. The ITP proved to be a powerful tool in assistance health care and an efficient strategy in the production of care.Estudo qualitativo, do tipo relato de experiência, de cunho crítico-reflexivo, sobre elaboração e desenvolvimento de um Projeto Terapêutico Singular (PTS) pela equipe Materno Infantil de um Programa de Residência Multiprofissional em Saúde, em um hospital público da região central do Rio Grande do Sul. O caso relatado descreve o acompanhamento, fundamentado no referencial teórico do PTS, de uma usuária do Sistema Único de Saúde, com diversas demandas biopsicossociais. O PTS iniciou-se durante internação da usuária na unidade tocoginecológica do hospital, campo de referência das residentes. Após o aceite da usuária em participar e da vinculação estabelecida entre ela e a equipe, realizou-se o planejamento das estratégias para atingir metas em curto, médio e longo prazos, que foram constantemente reavaliadas durante o período de acompanhamento. Como resultado da vivência, confirmou-se o que prima a literatura. O PTS mostra-se uma potente ferramenta na assistência integral à saúde e deve ser incentivado, não apenas como apoio à formação de residentes multiprofissionais, mas também entre as diferentes equipes de saúde da rede de atenção, como estratégia eficiente na produção do cuidado em saúde