Interferon-gamma alters the phagocytic activity of the mouse trophoblast

Interferon-gamma (IFN-gamma) mediates diverse functions in bone marrow-derived phagocytes, including phagocytosis and microbe destruction. This cytokine has also been detected at implantation sites under both physiological and pathological conditions in many different species. At these particular sites, the outermost embryonic cell layer in close contact with the maternal tissues, the trophoblast exhibits intense phagocytic activity. To determine whether IFN-gamma affects phagocytosis of mouse-trophoblast cells, ectoplacental cone-derived trophoblast was cultured and evaluated for erythrophagocytosis. Phagocytic activity was monitored ultrastructurally and expressed as percentage of phagocytic trophoblast in total trophoblast cells. Conditioned medium from concanavalin-A-stimulated spleen cells significantly enhanced trophoblast phagocytosis. This effect was blocked by pre-incubation with an anti-IFN-gamma neutralizing antibody. Introduction of mouse recombinant IFN-gamma (mrIFN-gamma) to cultures did not increase cell death, but augmented the percentage of phagocytic cells in a dose-dependent manner. Ectoplacental cones from mice deficient for IFN-gamma receptor alpha-chain showed a significant decrease of the phagocytosis, even under mrIFN-gamma stimulation, suggesting that IFN-gamma-induced phagocytosis are receptor-mediated. Reverse transcriptase-PCR analyses confirmed the presence of mRNA for IFN-gamma receptor alpha and beta-chains in trophoblast cells and detected a significant increase in the mRNA levels of IFN-gamma receptor beta-chain, mainly, when cultured cells were exposed to IFN-gamma. Immunohistochemistry and Western blot analyses also revealed protein expression of the IFN-gamma receptor alpha-chain. These results suggest that IFN-gamma may participate in the phagocytic activation of the mouse trophoblast, albeit the exact mechanism was not hereby elucidated. Protective and/or nutritional fetal benefit may result from this physiological response. In addition, our data also shed some light on the understanding of trophoblast tolerance to inflammatory/immune cytokines during normal gestation.

Two RND proteins involved in heavy metal efflux in Caulobacter crescentus belong to separate clusters within proteobacteria

Abstract Background Heavy metal Resistance-Nodulation-Division (HME-RND) efflux systems help Gram-negative bacteria to keep the intracellular homeostasis under high metal concentrations. These proteins constitute the cytoplasmic membrane channel of the tripartite RND transport systems. Caulobacter crescentus NA1000 possess two HME-RND proteins, and the aim of this work was to determine their involvement in the response to cadmium, zinc, cobalt and nickel, and to analyze the phylogenetic distribution and characteristic signatures of orthologs of these two proteins. Results Expression assays of the czrCBA operon showed significant induction in the presence of cadmium and zinc, and moderate induction by cobalt and nickel. The nczCBA operon is highly induced in the presence of nickel and cobalt, moderately induced by zinc and not induced by cadmium. Analysis of the resistance phenotype of mutant strains showed that the ΔczrA strain is highly sensitive to cadmium, zinc and cobalt, but resistant to nickel. The ΔnczA strain and the double mutant strain showed reduced growth in the presence of all metals tested. Phylogenetic analysis of the C. crescentus HME-RND proteins showed that CzrA-like proteins, in contrast to those similar to NczA, are almost exclusively found in the Alphaproteobacteria group, and the characteristic protein signatures of each group were highlighted. Conclusions The czrCBA efflux system is involved mainly in response to cadmium and zinc with a secondary role in response to cobalt. The nczCBA efflux system is involved mainly in response to nickel and cobalt, with a secondary role in response to cadmium and zinc. CzrA belongs to the HME2 subfamily, which is almost exclusively found in the Alphaproteobacteria group, as shown by phylogenetic analysis. NczA belongs to the HME1 subfamily which is more widespread among diverse Proteobacteria groups. Each of these subfamilies present distinctive amino acid signatures.

HOXB7 mRNA is overexpressed in pancreatic ductal adenocarcinomas and its knockdown induces cell cycle arrest and apoptosis

Background Human homeobox genes encode nuclear proteins that act as transcription factors involved in the control of differentiation and proliferation. Currently, the role of these genes in development and tumor progression has been extensively studied. Recently, increased expression of HOXB7 homeobox gene (HOXB7) in pancreatic ductal adenocarcinomas (PDAC) was shown to correlate with an invasive phenotype, lymph node metastasis and worse survival outcomes, but no influence on cell proliferation or viability was detected. In the present study, the effects arising from the knockdown of HOXB7 in PDAC cell lines was investigated. Methods Real time quantitative PCR (qRT-PCR) (Taqman) was employed to assess HOXB7 mRNA expression in 29 PDAC, 6 metastatic tissues, 24 peritumoral tissues and two PDAC cell lines. siRNA was used to knockdown HOXB7 mRNA in the cell lines and its consequences on apoptosis rate and cell proliferation were measured by flow cytometry and MTT assay respectively. Results Overexpression of HOXB7 mRNA was observed in the tumoral tissues and in the cell lines MIA PaCa-2 and Capan-1. HOXB7 knockdown elicited (1) an increase in the expression of the pro-apoptotic proteins BAX and BAD in both cell lines; (2) a decrease in the expression of the anti-apoptotic protein BCL-2 and in cyclin D1 and an increase in the number of apoptotic cells in the MIA PaCa-2 cell line; (3) accumulation of cell in sub-G1 phase in both cell lines; (4) the modulation of several biological processes, especially in MIA PaCa-2, such as proteasomal ubiquitin-dependent catabolic process and cell cycle. Conclusion The present study confirms the overexpression of HOXB7 mRNA expression in PDAC and demonstrates that decreasing its protein level by siRNA could significantly increase apoptosis and modulate several biological processes. HOXB7 might be a promising target for future therapies.

Laboratorial training of examiners for using a visual caries detection system in epidemiological surveys

Background: In epidemiological surveys, a good reliability among the examiners regarding the caries detection method is essential. However, training and calibrating those examiners is an arduous task because it involves several patients who are examined many times. To facilitate this step, we aimed to propose a laboratory methodology to simulate the examinations performed to detect caries lesions using the International Caries Detection and Assessment System (ICDAS) in epidemiological surveys. Methods: A benchmark examiner conducted all training sessions. A total of 67 exfoliated primary teeth, varying from sound to extensive cavitated, were set in seven arch models to simulate complete mouths in primary dentition. Sixteen examiners (graduate students) evaluated all surfaces of the teeth under illumination using buccal mirrors and ball-ended probe in two occasions, using only coronal primary caries scores of the ICDAS. As reference standard, two different examiners assessed the proximal surfaces by direct visual inspection, classifying them in sound, with non-cavitated or with cavitated lesions. After, teeth were sectioned in the bucco-lingual direction, and the examiners assessed the sections in stereomicroscope, classifying the occlusal and smooth surfaces according to lesion depth. Inter-examiner reproducibility was evaluated using weighted kappa. Sensitivities and specificities were calculated at two thresholds: all lesions and advanced lesions (cavitated lesions in proximal surfaces and lesions reaching the dentine in occlusal and smooth surfaces). Conclusion: The methodology purposed for training and calibration of several examiners designated for epidemiological surveys of dental caries in preschool children using the ICDAS is feasible, permitting the assessment of reliability and accuracy of the examiners previously to the survey´s development.

Functional markers for gene mapping and genetic diversity studies in sugarcane

Abstract Background The database of sugarcane expressed sequence tags (EST) offers a great opportunity for developing molecular markers that are directly associated with important agronomic traits. The development of new EST-SSR markers represents an important tool for genetic analysis. In sugarcane breeding programs, functional markers can be used to accelerate the process and select important agronomic traits, especially in the mapping of quantitative traits loci (QTL) and plant resistant pathogens or qualitative resistance loci (QRL). The aim of this work was to develop new simple sequence repeat (SSR) markers in sugarcane using the sugarcane expressed sequence tag (SUCEST database). Findings A total of 365 EST-SSR molecular markers with trinucleotide motifs were developed and evaluated in a collection of 18 genotypes of sugarcane (15 varieties and 3 species). In total, 287 of the EST-SSRs markers amplified fragments of the expected size and were polymorphic in the analyzed sugarcane varieties. The number of alleles ranged from 2-18, with an average of 6 alleles per locus, while polymorphism information content values ranged from 0.21-0.92, with an average of 0.69. The discrimination power was high for the majority of the EST-SSRs, with an average value of 0.80. Among the markers characterized in this study some have particular interest, those that are related to bacterial defense responses, generation of precursor metabolites and energy and those involved in carbohydrate metabolic process. Conclusions These EST-SSR markers presented in this work can be efficiently used for genetic mapping studies of segregating sugarcane populations. The high Polymorphism Information Content (PIC) and Discriminant Power (DP) presented facilitate the QTL identification and marker-assisted selection due the association with functional regions of the genome became an important tool for the sugarcane breeding program.

Cinnamic acid induces apoptotic cell death and cytoskeleton disruption in human melanoma cells

Anticancer activities of cinnamic acid derivatives include induction of apoptosis by irreversible DNA damage leading to cell death. The present work aimed to compare the cytotoxic and genotoxic potential of cinnamic acid in human melanoma cell line (HT-144) and human melanocyte cell line derived from blue nevus (NGM). Viability assay showed that the IC50 for HT-144 cells was 2.4 mM, while NGM cells were more resistant to the treatment. The growth inhibition was probably associated with DNA damage leading to DNA synthesis inhibition, as shown by BrdU incorporation assay, induction of nuclear aberrations and then apoptosis. The frequency of cell death caused by cinnamic acid was higher in HT-144 cells. Activated-caspase 3 staining showed apoptosis after 24 hours of treatment with cinnamic acid 3.2 mM in HT-144 cells, but not in NGM. We observed microtubules disorganization after cinnamic acid exposure, but this event and cell death seem to be independent according to M30 and tubulin labeling. The frequency of micronucleated HT-144 cells was higher after treatment with cinnamic acid (0.4 and 3.2 mM) when compared to the controls. Cinnamic acid 3.2 mM also increased the frequency of micronucleated NGM cells indicating genotoxic activity of the compound, but the effects were milder. Binucleation and multinucleation counting showed similar results. We conclude that cinnamic acid has effective antiproliferative activity against melanoma cells. However, the increased frequency of micronucleation in NGM cells warrants the possibility of genotoxicity and needs further investigation.

Doenças crônicas não transmissíveis no Brasil: prioridade para enfrentamento e investigação

As Doenças Crônicas Não Transmissíveis representam a maior carga de morbimortalidade no Brasil. Em 2011, o Ministério da Saúde lançou seu Plano de Ações Estratégicas para o Enfrentamento das Doenças Crônicas Não Transmissíveis, enfatizando ações populacionais para controlar as doenças cardiovasculares, diabetes, câncer e doença respiratória crônica, predominantemente pelo controle do fumo, inatividade física, alimentação inadequada e uso prejudicial de álcool. Apesar da produção científica significativa sobre essas doenças e seus fatores de risco no Brasil, poucos são os estudos de coorte nessa temática. Nesse contexto, o Estudo Longitudinal da Saúde do Adulto (ELSA-Brasil) acompanha 15.105 servidores públicos do País. Seus dados espelham a realidade brasileira de altas prevalências de diabetes e hipertensão e dos fatores de risco. A diversidade das informações produzidas permitirá aprofundar o entendimento causal dessas doenças e subsidiar políticas públicas para seu enfrentamento.

Ensino de enfermagem em centro cirúrgico: transformações da disciplina na Escola de Enfermagem da USP (Brasil)

Este artigo tem por objetivos apresentar uma síntese da evolução do conteúdo de bloco cirúrgico na Escola de Enfermagem da Universidade de São Paulo (EEUSP) e uma reflexão sobre as Diretrizes Curriculares Nacionais do Curso de Graduação em Enfermagem (DCN). O estudo se desenvolveu a partir de um breve histórico do desenvolvimento da enfermagem em centro cirúrgico na prática profissional e sua inserção no currículo de graduação da EEUSP. As Diretrizes Curriculares Nacionais têm seu mérito ao assegurar às instituições de ensino superior a liberdade na composição da carga horária a ser cumprida para a integralização dos currículos e na especificação das unidades de estudo a serem ministradas, porém, as competências e habilidades propostas são inespecíficas. Entendemos que o enfermeiro generalista é aquele que tem oportunidades de aprendizado teórico prático para atuar em todos os cenários de cuidado, área e níveis de atenção em saúde.

Association between particulate matter air pollution and monthly inhalation and nebulization procedures in Ribeirão Preto, São Paulo State, Brazil

The study was designed to investigate the impact of air pollution on monthly inhalation/nebulization procedures in Ribeirão Preto, São Paulo State, Brazil, from 2004 to 2010. To assess the relationship between the procedures and particulate matter (PM10) a Bayesian Poisson regression model was used, including a random factor that captured extra-Poisson variability between counts. Particulate matter was associated with the monthly number of inhalation/nebulization procedures, but the inclusion of covariates (temperature, precipitation, and season of the year) suggests a possible confounding effect. Although other studies have linked particulate matter to an increasing number of visits due to respiratory morbidity, the results of this study suggest that such associations should be interpreted with caution.

Custos da cadeia de procedimentos no tratamento do infarto agudo do miocárdio em hospitais brasileiros de excelência e especializados

OBJETIVO: Apresenta a metodologia e os resultados de uma pesquisa de campo para avaliar os custos da cadeia de procedimentos para tratamento do Infarto Agudo do Miocárdio (IAM), realizada em 11 hospitais brasileiros de excelência e especializados. MÉTODOS: A apuração do custo utilizou o sistema de custo por procedimento e o sistema por patologia. Os procedimentos associados ao tratamento do IAM foram organizados com sua sequência lógica (protocolos), dando origem a um fluxograma. Os instrumentos de coleta da pesquisa contemplavam informações relativas aos preços e as quantidades (particular, convênios, tabela SUS e Associação Médica Brasileira - AMB), praticados no ano de 2008, bem como os custos pertinentes. RESULTADOS: De forma geral, o custo total dos procedimentos que integram o "tratamento-padrão" do IAM, totalizou R$ 12.873,69 se a intervenção coronária percutânea (ICP) não envolver utilização de stent. Caso este se torne necessário, o custo se eleva para R$ 23.461,87. CONCLUSÃO: Entre os resultados obtidos destacam-se: o fato de os valores dos procedimentos mais caros não apresentarem variações estatísticas significativas entre os hospitais, independentemente de sua localização, clientela predominante ou natureza jurídica; e o fato de os hospitais, que tratam predominantemente de usuários do Sistema Único de Saúde, registrar custos menores, mas não estatisticamente significativos, para toda a cadeia de procedimentos associada à patologia.

Sem sol nem tempestade: caracteres de Iaiá Garcia em ruína

O artigo discute o desvio, realizado por Machado de Assis em Iaiá Garcia, das expectativas da narrativa sentimental e a comparação estabelecida entre os dois triângulos amorosos: Jorge, Estela e Luís Garcia, de um lado, e Estela, Jorge e Iaiá Garcia, de outro. Considerando a ação das paixões e do sistema escravocrata na conduta das personagens, aborda o confronto entre a convenção do amor paixão e a instituição do casamento como instrumento de ascensão social.

Manipulation of the periovulatory sex steroidal milieu affects endometrial but not luteal gene expression in early diestrus Nelore cows

In beef cattle, the ability to conceive has been associated positively with size of the preovulatory follicle (POF). Proestrus estradiol and subsequent progesterone concentrations can regulate the endometrium to affect receptivity and fertility. The aim of the present study was to verify the effect of the size of the POF on luteal and endometrial gene expression during subsequent early diestrus in beef cattle. Eighty-three multiparous, nonlactating, presynchronized Nelore cows received a progesterone-releasing device and estradiol benzoate on Day–10 (D 10). Animals received cloprostenol (large follicle-large CL group; LF-LCL; N ¼ 42) or not (small follicle-small CL group; SF-SCL; N ¼ 41) on D 10. Progesterone devices were withdrawn and cloprostenol administered 42 to 60 hours (LF-LCL) or 30 to 36 hours (SF-SCL) before GnRH treatment (D0). Tissues were collected at slaughter on D7. The LF-LCL group had larger (P < 0.0001) POF (13.24 0.33 mm vs. 10.76 0.29 mm), greater (P < 0.0007) estradiol concentrations on D0 (2.94 0.28 pg/mL vs. 1.27 0.20 pg/mL), and greater (P < 0.01) progesterone concentrations on D7 (3.71 0.25 ng/mL vs. 2.62 0.26 ng/mL) compared with the SF-SCL group. Luteal gene expression of vascular endothelial growth factor A, kinase insert domain receptor, fms-related tyrosine kinase 1, steroidogenic acute regulatory protein, cytochrome P450, family 11, subfamily A, polypeptide 1, and hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid deltaisomerase 7 was similar between groups. Endometrial gene expression of oxytocin receptor and peptidase inhibitor 3, skin-derived was reduced, and estrogen receptor alpha 2, aldo-keto reductase family 1, member C4, and lipoprotein lipase expression was increased in LF-LCL versus SF-SCL. Results support the hypothesis that the size of the POF alters the periovulatory endocrine milieu (i.e., proestrus estradiol and diestrus progesterone concentrations) and acts on the uterus to alter endometrial gene expression. It is proposed that the uterine environment and receptivity might also be modulated. Additionally, it is suggested that increased progesterone secretion of cows ovulating larger follicles is likely due to increased CL size rather than increased luteal expression of steroidogenic genes.

Horizontal distribution of invertebrate larvae around the oceanic island of Gran Canaria: the effect of mesoscale variability

In October 1991, the horizontal distribution of invertebrate larvae was studied in the waters surrounding the island of Gran Canaria (Canary Islands). The cruise was typified by the presence of three recurrent mesoscale hydrographic structures: a cyclonic eddy southwest of the island, a warm lee region downstream of the island and the offshore boundary of an upwelling filament from the African coast reaching the southeast of the island. Decapod larvae were the most abundant group. In general, a rather high spatial variability was found. The horizontal distribution of the invertebrate larvae groups showed that the highest values of abundance occurred in an elongated zone around the island oriented in the overall direction of flow, leeward and windward of the island, while the lowest values occurred off the eastern and western flanks of the islands. On the other hand, Stomatopoda and Mollusca larvae showed a distribution associated with the boundary of the upwelling filament and decapod larvae of pelagic species were distributed around the eddy structure. Our results suggest specific retention mechanisms for the larvae of neritic invertebrate populations that are related to the particular physical oceanography around Gran Canaria. Se estudió la distribución horizontal de las larvas de invertebrados alrededor de la isla de Gran Canaria (Islas Canarias) en octubre de 1991. La campaña se caracterizó por la presencia de tres fenómenos oceanográficos de mesoescala recurrentes: un remolino ciclónico al suroeste de la isla, la estela cálida a sotavento de la misma y el borde de un filamento proveniente del afloramiento africano situado al sureste de Gran Canaria. Las larvas de crustáceos decápodos fueron las más abundantes. En general, las larvas de los diferentes taxa estudiados mostraron una gran variabilidad en cuanto a su distribución espacial. La distribución horizontal de las larvas de los diferentes grupos de invertebrados mostró que las densidades más altas se registraban alrededor de la isla en la banda cercana y orientada hacia el suroeste, en la dirección del flujo, mientras que los valores más bajos se encontraron en el norte y en los flancos este y oeste de la isla. Por otro lado, las larvas de estomatópodos y moluscos presentaron una distribución asociada al frente del filamento, mientras que las larvas de decápodos pelágicos se distribuyeron en torno al remolino ciclónico. Los resultados obtenidos sugieren la existencia de mecanismos de retención específicos para el mantenimiento de las poblaciones insulares, relacionados con los procesos físicos de mesoescala predominantes

Temporal and spatial distribution of the ichthyoplankton in the Canary Islands

A pesar del creciente interés en el estudio de las primeras fases de desarrollo de los peces, aún existen zonas donde esta comunidad es relativamente desconocida. Este es el caso de las Islas Canarias. En la presente tesis, gracias a una serie de muestreos semanales en la región costera de la isla de Gran Canaria durante más de dos años, se ha caracterizado de manera precisa la composición, estructura y variabilidad del ictioplancton presente en esta región subtropical. Clupeidos, Espáridos y Góbidos dominaron la comunidad neríticas, mientras que Mictófidos, Gonostomátidos y Photíctidos prevalecieron dentro de las familias oceánicas. La variabilidad temporal de la comunidad larvaria parece estar más relacionada con la temperatura y efectos a pequeña escala (hidrografía, productividad local, ciclo lunar), que con el aumento de productividad anual durante el bloom de finales de invierno. Se definieron dos asociaciones estacionales de larvas estrechamente ligadas a las características de la columna de agua: (1) invierno-primavera, en la que dominan Sardinella aurita, Boops boops y Cyclothone braueri, y son características especies como Pomacentridae sp1, Trachurus picturatus o Scomber colias; y (2) verano-otoño, donde Góbidos y Cyclothone braueri dominan, pero cuyas especies características son Ceratoscopelus warmingii, Pomacentridae sp2 y Anthias anthias, entre otras. Respecto a la variabilidad horizontal, se confirmó la presencia de dos zonas de retención para los huevos y larvas neríticas a barlovento y sotavento (estela cálida) de la isla. Sin embargo, no se encontraron diferencias significativas en la composición de la comunidad larvaria entre las diferentes zonas de la plataforma de la isla. La Zona de Transición Costera Canario-Africana se caracteriza por una gran actividad de mesoscala, que afectará a la distribución de las larvas neríticas de las costas africanas. Esta interacción resulta especialmente evidente en el caso de las larvas, principalmente sardina y anchoa, transportadas en filamentos de afloramiento generados en la región de Cabo Juby-Cabo Bojador. Estos filamentos pueden ser atrapados por remolinos ciclónicos situados al sur de Gran Canaria. Este sistema remolino-filamento podrá, en función de su evolución, actuar como un mecanismo de retención o dispersión para las larvas de especies neríticas africanas. Parte de estas larvas pueden llegar a las costas de Gran Canaria, suponiendo un aporte para las poblaciones larvarias locales, al menos durante el verano. A pesar de la gran cantidad de información obtenida a partir de esta tesis, es necesario continuar con este tipo de estudios para evaluar la importancia de este transporte larvario y su aplicación a la gestión pesquera.