Identifying genes related to choriogenesis in insect panoistic ovaries by Suppression Subtractive Hybridization

<p>Abstract</p> <p>Background</p> <p>Insect ovarioles are classified into two categories: panoistic and meroistic, the later having apparently evolved from an ancestral panoistic type. Molecular data on oogenesis is practically restricted to meroistic ovaries. If we aim at studying the evolutionary transition from panoistic to meroistic, data on panoistic ovaries should be gathered. To this end, we planned the construction of a Suppression Subtractive Hybridization (SSH) library to identify genes involved in panoistic choriogenesis, using the cockroach <it>Blattella germanica </it>as model.</p> <p>Results</p> <p>We constructed a post-vitellogenic ovary library by SSH to isolate genes involved in choriogenesis in <it>B. germanica</it>. The tester library was prepared with an ovary pool from 6- to 7-day-old females, whereas the driver library was prepared with an ovary pool from 3- to 4-day-old females. From the SSH library, we obtained 258 high quality sequences which clustered into 34 unique sequences grouped in 19 contigs and 15 singlets. The sequences were compared against non-redundant NCBI databases using BLAST. We found that 44% of the unique sequences had homologous sequences in known genes of other organisms, whereas 56% had no significant similarity to any of the databases entries. A Gene Ontology analysis was carried out, classifying the 34 sequences into different functional categories. Seven of these gene sequences, representative of different categories and processes, were chosen to perform expression studies during the first gonadotrophic cycle by real-time PCR. Results showed that they were mainly expressed during post-vitellogenesis, which validates the SSH technique. In two of them corresponding to novel genes, we demonstrated that they are specifically expressed in the cytoplasm of follicular cells in basal oocytes at the time of choriogenesis.</p> <p>Conclusion</p> <p>The SSH approach has proven to be useful in identifying ovarian genes expressed after vitellogenesis in <it>B. germanica</it>. For most of the genes, functions related to choriogenesis are postulated. The relatively high percentage of novel genes obtained and the practical absence of chorion genes typical of meroistic ovaries suggest that mechanisms regulating chorion formation in panoistic ovaries are significantly different from those of meroistic ones.</p

Brownie, a Gene Involved in Building Complex Respiratory Devices in Insect Eggshells

Background: Insect eggshells must combine protection for the yolk and embryo with provisions for respiration and for the entry of sperm, which are ensured by aeropyles and micropyles, respectively. Insects which oviposit the eggs in an egg-case have a double problem of respiration as gas exchange then involves two barriers. An example of this situation is found in the cockroach Blattella germanica, where the aeropyle and the micropyle are combined in a complex structure called the sponge-like body. The sponge-like body has been well described morphologically, but nothing is known about how it is built up. Methodology/Principal Findings: In a library designed to find genes expressed during late chorion formation in B. germanica, we isolated the novel sequence Bg30009 (now called Brownie), which was outstanding due to its high copy number. In the present work, we show that Brownie is expressed in the follicle cells localized in the anterior pole of the oocyte in late choriogenesis. RNA interference (RNAi) of Brownie impaired correct formation of the sponge-like body and, as a result, the egg-case was also ill-formed and the eggs were not viable. Conclusions/Significance: Results indicate that the novel gene Brownie plays a pivotal role in building up the sponge-lik

Preventive Gambling Programs for Adolescents and Young Adults: A Systematic Review

Gambling disorder in youth is an emerging public health problem, with adolescents and young adults constituting a vulnerable age group for the development of gambling-related problems. Although research has been conducted on the risk factors for gambling disorder, very few rigorous studies can be found on the efficacy of preventive interventions in young people. The aim of this study was to provide best practice recommendations for the prevention of disordered gambling in adolescents and young adults. We reviewed and synthesized the results of existing RCTs and quasi-experimental studies covering nonpharmacological prevention programs for gambling disorder in young adults and adolescents. We applied the PRISMA 2020 statement and guidelines to identify 1483 studies, of which 32 were included in the systematic review. All studies targeted the educational setting, i.e., high school and university students. Most studies followed a universal prevention strategy, that particularly targeted adolescents, and an indicated prevention strategy for university students. The reviewed gambling prevention programs generally showed good results in terms of reducing the frequency and severity of gambling, and also regarding cognitive variables, such as misconceptions, fallacies, knowledge, and attitudes towards gambling. Finally, we highlight the need to develop more comprehensive prevention programs that incorporate rigorous methodological and assessment procedures before they are widely implemented and disseminated

The Changing Landscape for Stroke\ua0Prevention in AF: Findings From the GLORIA-AF Registry Phase 2

Background GLORIA-AF (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients with Atrial Fibrillation) is a prospective, global registry program describing antithrombotic treatment patterns in patients with newly diagnosed nonvalvular atrial fibrillation at risk of stroke. Phase 2 began when dabigatran, the first non\u2013vitamin K antagonist oral anticoagulant (NOAC), became available. Objectives This study sought to describe phase 2 baseline data and compare these with the pre-NOAC era collected during phase&nbsp;1. Methods During phase 2, 15,641 consenting patients were enrolled (November 2011 to December 2014); 15,092 were eligible. This pre-specified cross-sectional analysis describes eligible patients\u2019 baseline characteristics. Atrial fibrillation&nbsp;disease characteristics, medical outcomes, and concomitant diseases and medications were collected. Data were analyzed using descriptive statistics. Results Of the total patients, 45.5% were female; median age was 71 (interquartile range: 64, 78) years. Patients were from Europe (47.1%), North America (22.5%), Asia (20.3%), Latin America (6.0%), and the Middle East/Africa (4.0%). Most had high stroke risk (CHA2DS2-VASc [Congestive heart failure, Hypertension, Age&nbsp; 6575 years, Diabetes mellitus, previous Stroke, Vascular disease, Age 65 to 74 years, Sex category] score&nbsp; 652; 86.1%); 13.9% had moderate risk (CHA2DS2-VASc&nbsp;= 1). Overall, 79.9% received oral anticoagulants, of whom 47.6% received NOAC and 32.3% vitamin K antagonists (VKA); 12.1% received antiplatelet agents; 7.8% received no antithrombotic treatment. For comparison, the proportion of phase 1 patients (of N&nbsp;= 1,063 all eligible) prescribed VKA was 32.8%, acetylsalicylic acid 41.7%, and no therapy 20.2%. In Europe in phase 2, treatment with NOAC was more common than VKA (52.3% and 37.8%, respectively); 6.0% of patients received antiplatelet treatment; and 3.8% received no antithrombotic treatment. In North America, 52.1%, 26.2%, and 14.0% of patients received NOAC, VKA, and antiplatelet drugs, respectively; 7.5% received no antithrombotic treatment. NOAC use was less common in Asia (27.7%), where 27.5% of patients received VKA, 25.0% antiplatelet drugs, and 19.8% no antithrombotic treatment. Conclusions The baseline data from GLORIA-AF phase 2 demonstrate that in newly diagnosed nonvalvular atrial fibrillation patients, NOAC have been highly adopted into practice, becoming more frequently prescribed than VKA in&nbsp;Europe and North America. Worldwide, however, a large proportion of patients remain undertreated, particularly in&nbsp;Asia&nbsp;and North America. (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients With Atrial Fibrillation [GLORIA-AF]; NCT01468701

Os aspectos semiológicos do acidente vascular encefálico: uma abordagem neurológica

O Acidente Vascular Encefálico (AVC) é um evento neurológico súbito, com um foco de isquemia ou hemorragia. Ambos, qualificados pelo déficit neurológico focal abrupto. Ressaltando, que estes déficits podem ocorrer, sendo a ocorrência espontânea, perduração de 15 minutos, autoresolutiva é denominada como Ataque Isquêmico Transitório (AIT), no entanto, toda insuficiência neural que não melhorar pós esse período deve ser manejado como AVC. O artigo objetivou descrever os principais aspectos clínicos do AVC. O AVC é uma emergência para a saúde pública, em razão de ser um potencial em gerar morbimortalidades para os portadores e prejuízos para os sistemas de saúde. O AVC do tipo isquêmico representa a maioria das ocorrências, o quadro clínico do paciente é correspondente ao tecido neural afetado, inicialmente a tomografia computadorizada sem contraste é o primeiro exame, por ser crucial para descartar a etiologia hemorrágica, a condução terapêutica se baseia em medidas neuroprotetoras através da estabilização da glicemia, temperatura e sódio, adequar os níveis pressóricos, mediante o prazo estipulado impor terapia antitrombótica. A manifestação hemorrágica, pode ocorrer por torção de aneurisma sacular originando o sangramento subaracnóideo ou por hipertensão gerando o sangramento intraparenquimatoso. A partir da análise das informações coletadas, elucida-se que o diagnóstico precoce e o período transcorrido até o manejo terapêutico são cruciais para o desfecho clínico do portador, ou seja, é possível a normalização ou ocorrer sequelas neurais e óbito

RNAi reveals the key role of Nervana 1 in cockroach oogenesis and embryo development

Na+, K+-ATPases is a heterodimer protein consisting of α- and β-subunits that control the ion transport through cell membranes. In insects the β-subunit of the Na+, K+-ATPase, known as Nervana, was characterized as a nervous system-specific glycoprotein antigen from adult Drosophila melanogaster heads. Nervana is expressed ubiquitously in all insect tissues, and in epithelial cells appeared located in a basolateral position as part of the septate junctions. Herein we study two Nervana isoforms from Blattella germanica, a cockroach species with panoistic ovaries. The sequencing and the phylogenetic analysis results suggest that these two isoforms are orthologs of D. melanogaster Nervana 1 and Nervana 2, respectively. Nervana 1 is highly expressed in the ovary of B. germanica, and depleting its expression results in changes in oocyte shape that do not impair oviposition. However, the resulting embryos show different defects and never hatch. These findings highlight the importance of this type of membrane pump in insect oogenesis as well as in embryo development, and its possible regulation by juvenile hormone. © 2012 Elsevier Ltd.This work was supported by the Ministry of Science and Innovation, Spain (projects BFU2008-00484 and BFU2011-22404 to MDP), and by the CSIC (grant 2010TW0019, Formosa program). PI is recipient of a postdoctoral research grant (BECAS CHILE) from the “Comisión Nacional de Investigación Científica y Tecnológica” (CONICYT, Chile). FAST was supported by a fellowship from FAPESP, to do a stage in the IBE.Peer Reviewe

The Hippo pathway in oogenesis of Blattella germanica

Trabajo presentado en el XXIV International Congress of Entomology "New era in Entomology" (ICE 2012), celebrado en Daegu (Corea del Sur) del 19 al 25 de agosto de 2012.N

Yorkie/YAP: The transcription factor effector of Hippo pathway unveils a novel function as an activator of growth during insect oogenesis.

Trabajo presentado en la XXIX Annual Meeting Chilean Society for Cell Biology, celebrada en Puerto Varas (Chile) del 25 al 29 de octubre de 2015.N

Deciphering the action and interaction of cellular signaling pathways, Hippo and Notch during ovarian development in insects

Trabajo presentado en la LVIII Reunion Anual Sociedad Biológica de Chile y XIII Reunión Anual Sociedad Chilena de Inmunología, celebrado en Puerto Varas (Chile) del 23 al 25 de noviembre de 20153El éxito reproductivo de una especie depende, en gran parte, de los eventos asociados al proceso de maduración ovárica que involucra desde la definición de las células germinales primordiales hasta el establecimiento de la polaridad del embrión. Nuestro interés se ha centrado en entender cuáles son los mecanismos moleculares involucrados en la oogénesis de insectos basales. Dentro de los objetivos se encuentra el estudio de las vías de señalización celular de Hippo y Notch, las cuales determinan el correcto balance entre procesos de proliferación, diferenciación y muerte celular. Así, hemos determinado los patrones de expresión y localización de los principales componentes de estas vías. Además, usando el RNA de interferencia (RNAi) para el silenciamiento génico, hemos descrito funcionalmente las proteínas de Hippo, Yorkie y Notch en el ovario primitivo de la cucaracha Blattella germanica . Los resultados demuestran que la función supresora de tumores de la vía Hippo se encuentra conservada a lo largo de la escala evolutiva. Sin embargo, la interacción entre las dos vías es diferente: en oposición a la mosca de la fruta, Drosophila melanogaster , Hippo reprime Notch y gatilla la transición desde el estado mitótico de las células foliculares al endociclo, donde Yorkie tiene un papel fundamental. Los resultados sugieren que este rol de Hippo correspondería al más ancestral. En la actualidad hemos introducido un nuevo insecto en nuestros estudios, la tijereta Euborellia annulipes , que posee un ovario que representaría la transición estructural y funcional entre los dos tipos de ovarios encontrados en insectos. Los resultados obtenidos permitirán consensuar los mecanismos de acción y de interacción entre las principales vías de señalización celular en el ovario que podrían ser extensibles al resto de artrópodos. Finalmente, se podrán identificar los elementos claves que operan en el correcto balance entre eventos celulares proliferativos y apoptóticos.El trabajo ha sido financiado por el Ministerio de Ciencia e Innovación, España (BFU2011-22404). Beca postdoctoral BECAS CHILE, CONICYT y Programa Apoyo al Retorno de Doctores desde el Extranjero (821320046), Convocatoria 2013, PAI, CONICYT.N

The hippo pathway controls cell proliferation in insect oogenesis

Trabajo presentado en el 9th Congress of the Asociación Ibérica de Endocrinología Comparada (AIEC) y 17th International Congress of Comparative Endocrinology (ICCE 2013), celebrados en Barcelona en julio de 2013.N