Simulação in Situ, uma Metodologia de Treinamento Multidisciplinar para Identificar Oportunidades de Melhoria na Segurança do Paciente em uma Unidade de Alto Risco

Introdução A simulação é uma metodologia usada para substituir ou amplificar experiências reais por experiências guiadas que evocam ou replicam aspectos do mundo real de maneira interativa. A simulação in situ leva essa técnica diretamente aos locais onde ocorrem atendimentos, com a própria equipe de saúde atuando em seu ambiente de trabalho em cenário simulado. Objetivo Descrever experiência piloto de simulação in situ realizada em unidade de pronto atendimento, destacando oportunidades de avaliação de sistema de atendimento, trabalho em equipe e detecção de ameaças latentes à segurança (ALS). Métodos Estudo aplicado na Unidade Ibirapuera do Hospital Israelita Albert Einstein e realizado pelo Centro de Simulação Realística Albert Einstein. Foi apresentado cenário de paciente de 45 anos com síndrome coronariana aguda que evolui para parada cardiorrespiratória. Simulação híbrida de 30 minutos com uso de ator e simulador de alta fidelidade (SimMan 3G®).Utilizado checklist e filmagem para avaliar habilidades e atitudes, usados em debriefing estruturado com uma hora de duração. Resultados A experiência proporcionou avaliação técnica, comportamental e sistemas. Detectou quatro ALS e permitiu reflexão guiada sobre trabalho em equipe. Conclusão Este piloto contribuiu para o alcance dos objetivos propostos com o cenário e demonstrou oportunidades de treinamento e melhoria. A simulação in situ pode ser usada no futuro sistematicamente para treinamento contínuo de equipes, visando à melhoria da qualidade de atendimento e à segurança do paciente.

Interceptação das chuvas em um fragmento de floresta da Mata Atlântica na Bacia do Prata, Recife, PE

A mata de Dois Irmãos é uma das poucas áreas remanescentes da Floresta Atlântica no Estado de Pernambuco. Nela estão inseridos os açudes do Meio, do Prata e Dois Irmãos que compõem a bacia hidrográfica do Prata. Este trabalho teve como objetivo estudar a partição das chuvas em um fragmento de Floresta Atlântica na Bacia do Prata em Recife, PE. Para obtenção dos dados de precipitação sob florestas, foram instalados 24 interceptômetros e selecionadas 20 árvores do estrato superior, e 10 árvores do sub-bosque foram escolhidas para obter os dados de escoamento pelo tronco. Encontraram-se perdas por interceptação de 208,3 mm, precipitação efetiva de 1.431,7 mm, precipitação interna de 1.392,4 mm, escoamento pelo tronco das árvores do estrato superior de 6,6 mm e escoamento pelo sub-bosque de 32,8 mm, correspondendo a 12,7%, 87,3%, 84,9%, 0,4% e 2%, respectivamente, do total precipitado de 1.464 mm.

Chromosome number and secondary constriction variation in 51 accessions of a citrus germplasm bank

The mitotic chromosomes of 51 citrus accessions from the Centro Nacional de Pesquisa em Mandioca e Fruticultura Tropical, Cruz das Almas, BA, Brazil, were analyzed. The sample included representatives of 20 Citrus species, one of Poncirus and seven hybrids. All accessions showed 2n = 18 without any evidence of numerical variation. The most clearly variable karyotype feature was the number and position of secondary constrictions (SECs). In 19 accessions the SECs were not identified, mainly due to the degree of chromatin condensation. In the remainder they varied in number from one to three per karyotype. They were found in the proximal region of one of the three largest chromosome pairs, in the terminal/subterminal region of a smaller chromosome or, more seldom, terminally in a larger chromosome. Only in a few cases were such constrictions observed simultaneously in both homologues of the same chromosome pair. The high variability of this karyotype feature may be due to the activation of this region in the previous interphase but may also indicate a high structural variability and heterozygosity of citrus germplasms

Concanavalin A-reactive nuclear matrix glycoprotein

The binding capacity of concanavalin A (Con A) to condensed euchromatin and heterochromatin was investigated in chicken erythrocyte nuclei (CEN), mouse liver cells, Zea mays mays meristematic cells and Drosophila melanogaster polytene chromosomes after 4 N HCl hydrolysis to determine whether binding was preferentially occurring in bands and heterochromatin. Dry mass (DM) variation was investigated in CEN by interference microscopy. Feulgen and Con A reactions were employed for all materials to correlate the loci of the two reactions. Quantifications and topological verifications were carried out by video image analysis (high performance cytometry). It was observed that 4 N HCl hydrolysis caused an important DM loss in CEN leaving a level corresponding to the average DNA DM content. In this material, Con A binding was restricted to the nuclear envelope, which reinforces the idea of the absence of a nuclear matrix in these cells. The other cell types exhibited a correspondence of Feulgen-positive and Con A-reactive areas. The Con A reaction was highly positive in the condensed chromatin areas and heterochromatin. This fact led us to speculate that Con A-positive proteins may play a role in the chromatin condensation mechanism, endowing this structure with physico-chemical stability towards acid hydrolysis and contributing to its rheological properties.

Induction of apoptosis in cancer cells by tumor necrosis factor and butyrolactone, an inhibitor of cyclin-dependent kinases

Induction of apoptosis by tumor necrosis factor (TNF) is modulated by changes in the expression and activity of several cell cycle regulatory proteins. We examined the effects of TNF (1-100 ng/ml) and butyrolactone I (100 µM), a specific inhibitor of cyclin-dependent kinases (CDK) with high selectivity for CDK-1 and CDK-2, on three different cancer cell lines: WEHI, L929 and HeLa S3. Both compounds blocked cell growth, but only TNF induced the common events of apoptosis, i.e., chromatin condensation and ladder pattern of DNA fragmentation in these cell lines. The TNF-induced apoptosis events were increased in the presence of butyrolactone. In vitro phosphorylation assays for exogenous histone H1 and endogenous retinoblastoma protein (pRb) in the total cell lysates showed that treatment with both TNF and butyrolactone inhibited the histone H1 kinase (WEHI, L929 and HeLa) and pRb kinase (WEHI) activities of CDKs, as compared with the controls. The role of proteases in the TNF and butyrolactone-induced apoptosis was evaluated by comparing the number and expression of polypeptides in the cell lysates by gel electrophoresis. TNF and butyrolactone treatment caused the disappearance of several cellular protein bands in the region between 40-200 kDa, and the 110- 90- and 50-kDa proteins were identified as the major substrates, whose degradation was remarkably increased by the treatments. Interestingly, the loss of several cellular protein bands was associated with the marked accumulation of two proteins apparently of 60 and 70 kDa, which may be cleavage products of one or more proteins. These findings link the decrease of cyclin-dependent kinase activities to the increase of protease activities within the growth arrest and apoptosis pathways induced by TNF.

Cholesterol induces fetal rat enterocyte death in culture

The effect of cholesterol on fetal rat enterocytes and IEC-6 cells (line originated from normal rat small intestine) was examined. Both cells were cultured in the presence of 20 to 80 µM cholesterol for up to 72 h. Apoptosis was determined by flow cytometric analysis and fluorescence microscopy. The expression of HMG-CoA reductase and peroxisome proliferator-activated receptor gamma (PPARgamma) was measured by RT-PCR. The addition of 20 µM cholesterol reduced enterocyte proliferation as early as 6 h of culture. Reduction of enterocyte proliferation by 28 and 41% was observed after 24 h of culture in the presence and absence of 10% fetal calf serum, respectively, with the effect lasting up to 72 h. Treatment of IEC-6 cells with cholesterol for 24 h raised the proportion of cells with fragmented DNA by 9.7% at 40 µM and by 20.8% at 80 µM. When the culture period was extended to 48 h, the effect of cholesterol was still more pronounced, with the percent of cells with fragmented DNA reaching 53.5% for 40 µM and 84.3% for 80 µM. Chromatin condensation of IEC-6 cells was observed after treatment with cholesterol even at 20 µM. Cholesterol did not affect HMG-CoA reductase expression. A dose-dependent increase in PPARgamma expression in fetal rat enterocytes was observed. The expression of PPAR-gamma was raised by 7- and 40-fold, in the presence and absence of fetal calf serum, respectively, with cholesterol at 80 mM. The apoptotic effect of cholesterol on enterocytes was possibly due to an increase in PPARgamma expression.

Symmetric pollen mitosis I and suppression of pollen mitosis II prevent pollen development in Brachiaria jubata (Gramineae)

Microsporogenesis and pollen development were analyzed in a tetraploid (2n = 4x = 36) accession of the forage grass Brachiaria jubata (BRA 007820) from the Embrapa Beef Cattle Brachiaria collection that showed partial male sterility. Microsporocytes and pollen grains were prepared by squashing and staining with 0.5% propionic carmine. The meiotic process was typical of polyploids, with precocious chromosome migration to the poles and laggards in both meiosis I and II, resulting in tetrads with micronuclei in some microspores. After callose dissolution, microspores were released into the anther locule and appeared to be normal. Although each microspore initiated its differentiation into a pollen grain, in 11.1% of them nucleus polarization was not observed, i.e., pollen mitosis I was symmetric and the typical hemispherical cell plate was not detected. After a central cytokinesis, two equal-sized cells showing equal chromatin condensation and the same nuclear shape and size were formed. Generative cells and vegetative cells could not be distinguished. These cells did not undergo the second pollen mitosis and after completion of pollen wall synthesis each gave rise to a sterile and uninucleate pollen grain. The frequency of abnormal pollen mitosis varied among flowers and also among inflorescences. All plants were equally affected. The absence of fertile sperm cells in a considerable amount of pollen grains in this accession of B. jubata may compromise its use in breeding and could explain, at least in part, why seed production is low when compared with the amount of flowers per raceme.

CCN2/CTGF silencing blocks cell aggregation in embryonal carcinoma P19 cell

Connective tissue growth factor (CCN2/CTGF) is a matricellular-secreted protein involved in extracellular matrix remodeling. The P19 cell line is an embryonic carcinoma line widely used as a cellular model for differentiation and migration studies. In the present study, we employed an exogenous source of CCN2 and small interference RNA to address the role of CCN2 in the P19 cell aggregation phenomenon. Our data showed that increasing CCN2 protein concentrations from 0.1 to 20 nM decreased the number of cell clusters and dramatically increased cluster size without changing proliferation or cell survival, suggesting that CCN2 induced aggregation. In addition, CCN2 specific silencing inhibited typical P19 cell aggregation, which could be partially rescued by 20 nM CCN2. The present study demonstrates that CCN2 is a key molecule for cell aggregation of embryonic P19 cells.

A phthalide derivative isolated from endophytic fungiPestalotiopsis photiniae induces G1 cell cycle arrest and apoptosis in human HeLa cells

MP [4-(3′,3′-dimethylallyloxy)-5-methyl-6-methoxyphthalide] was obtained from liquid culture of Pestalotiopsis photiniaeisolated from the Chinese Podocarpaceae plant Podocarpus macrophyllus. MP significantly inhibited the proliferation of HeLa tumor cell lines. After treatment with MP, characteristic apoptotic features such as DNA fragmentation and chromatin condensation were observed in DAPI-stained HeLa cells. Flow cytometry showed that MP induced G1 cell cycle arrest and apoptosis in a dose-dependent manner. Western blotting and real-time reverse transcription-polymerase chain reaction were used to investigate protein and mRNA expression. MP caused significant cell cycle arrest by upregulating the cyclin-dependent kinase inhibitor p27KIP1 protein and p21CIP1 mRNA levels in HeLa cells. The expression of p73 protein was increased after treatment with various MP concentrations. mRNA expression of the cell cycle-related genes, p21CIP1, p16INK4a and Gadd45α, was significantly upregulated and mRNA levels demonstrated significantly increased translation ofp73, JunB, FKHR, andBim. The results indicate that MP may be a potential treatment for cervical cancer.

Quality of minimally processed guava with different types of cut, sanification and packing

The purpose of this project was to evaluate the sanitization effect on the quality of minimally processed guava. Initially, research was carried out with consumers in a supermarket to verify preferences of packaging for guava. Following this, the guava cv. Paluma underwent two sanitization sequences using dehydrated sodium dichloroisocyanurate compound, in 50 ppm concentration, sanitization prior to (S1) and after (S2) being cut; removal of excess water; conditioning in PET packaging and PSPVC and storage at 3 ºC ± 1 ºC. Physicochemical analysis - [pH, total soluble solids (SST), total labeled acidity (ATT), ascorbic acid (AA), total sugars (AT) and reducers (AR)], textural sensorial and microbiological analyses were used to monitor the quality of the products. The consumers preferred the guava cut in halves with pulp and packed in PET, although this packaging promoted condensation of water vapor on the inner surface of the lid, compromising the appearance of the product. The two sanitization sequences and the two kinds of packaging did not significantly affect the pH, SST, ATT, SST/ATT, texture and AA values. The AT and AR tenors increased significantly in the MP guavas stored in the PSPVC package. Both sanitizations were efficient in the bacterial control of the indicators of the hygienicsanitary conditions, although the S1 sanitization proved to be more efficient in the control of autochthonous aerobic microbiota (aerobic mesophylic microorganisms). It can be concluded that guava cv. Paluma packed in PSPVC can be conserved for 6 days when stored at 3 ºC.