Prologica e a política nacional de informática

Este artigo tem por finalidade falar sobre a Prologica Computadores, uma das maiores empresas de fabricação de microcomputadores e periféricos dos anos 80, que teve seu auge durante a Reserva de Mercado Brasileira (Lei Federal nº 7.232/84), seus principais produtos, sua P&D, sua engenharia reversa e toda sua estrutura de várias empresas complementares

Avaliação preliminar da tecnologia "MI-Dengue" para o monitoramento e controle do Aedes aegypti

The "Intelligent Dengue Monitoring" technology (MI-Dengue) consists of a trap that captures gravid female Aedes aegypti, and is coupled with a computerized system for field data collection, transmission, and access to georeferenced maps in real time. This study describes the first experience with a system for monitoring adult Aedes aegypti. It presents the preliminary results in the three municipalities that adopted MI-Dengue as a strategy to identify key areas and orient control measures. Preliminary results suggest that this control strategy combined with house-to-house visits in a 200m radius of the trap helped reduce dengue in the municipalities that adopted the system.As limitações na identificação do Aedes aegypti em laboratório e no processamento das informações obtidas em campo pelo método da pesquisa larvária levaram ao desenvolvimento do "Monitoramento Inteligente da Dengue" (MI-Dengue). O MI-Dengue consiste em uma armadilha que captura fêmeas grávidas de Ae. aegypti associada ao sistema informatizado de coleta, transmissão e acesso das informações de campo, e mapas georreferenciados em tempo real. O objetivo deste trabalho foi descrever pela primeira vez um sistema de monitoramento de adultos de Ae. aegypti e apresentar os resultados preliminares em três municípios que adotaram o MI-Dengue como estratégia para identificar áreas e direcionar as ações de controle. Semanalmente, mapas georreferenciados e o indicador entomológico (IMFA) forneceram informações das áreas onde os níveis de infestações, caracterizados por cores em função da quantidade de fêmeas de Ae. aegypti capturadas, indicaram situação de sem risco, alerta e crítica que desencadearam ações de controle. Os resultados preliminares sugerem que a adoção dessa estratégia de controle com visitas casa a casa em um raio de 200m da armadilha positiva contribuiu para a redução de casos de dengue nos municípios que adotaram o MI-Dengue

New Cost-Benefit of Brazilian Technology for Vector Surveillance Using Trapping System

The recent introduction of chikungunya and Zika virus and their subsequent dispersion in the Americas have encouraged the use of novel technologies for adult Aedes surveillance to improve vector control. In Brazil, two platforms for surveillance of eggs and gravid Aedes aegypti have been developed. First, it consists of using data of sampling of eggs in ovitraps associated with GIS technologies to monitor Aedes spp. populations. Although effective, it is not realistic to use in a large-scale epidemic scenario as it requires a large amount of human resources for field and laboratory activities. Second, it consists of trapping female Ae. aegypti citywide at fine spatial and temporal scales for vector surveillance (MI-Aedes) to detect high Aedes infestation areas using a GIS environment and the identification of arbovirus-infected trapped mosquitoes by RT-PCR (MI-Virus platforms). Such integration of continuous vector surveillance and targeting vector control in hotspot areas is cost-effective (less than US$ 1.00/person/year), and it has been shown to reduce mosquito population and prevent dengue transmission. The main advantage of the MI-Aedes platform over traditional mosquito surveillance is the integration of continuous vector monitoring coupled with an information technology platform for near real-time data collection, analysis, and decision-making. The technologies also provide data to model the role of climate on the vector population dynamics

Viroids in Citrus

Os viroides são os menores fitopatógenos conhecidos. Constituídos por uma molécula de RNA de fita simples, circular, que não é encapsidada e não codifica proteínas, são capazes de se replicar de maneira autônoma nas células do hospedeiro. Os viroides de citros pertencem à família Pospiviroidae (cujos membros apresentam uma região central conservada, replicam-se no núcleo das células hospedeiras e não apresentam atividade ribozimática) com cinco espécies: Citrus exocortis viroid, CEVd (Pospiviroid), Hop stunt viroid, HSVd (Hostuviroid), Citrus bark cracking viroid, CBCVd (Cocadviroid) e Citrus bent leaf viroid, CBLVd e Citrus dwarfing viroid, CDVd (Apscaviroid). Além disso, Citrus viroid original source (CVd-OS) e mais recentemente, Citrus viroid V (CVd-V) foram propostas como espécies tentativas do gênero Apscaviroid. Os viroides de citros são transmitidos via enxertia e disseminados principalmente pela propagação de material contaminado. Sabe-se que os viroides de citros infectam praticamente todas as espécies do gênero Citrus e afins. Porém, há somente duas doenças importantes descritas em citros, induzidas por viroides: (i) a exocorte; (ii) e a xiloporose (ou cachexia). Apesar dos viroides induzirem sintomas severos, ou simplesmente afetarem o tamanho das árvores, muitas espécies de citros são assintomáticas, sendo o controle baseado em medidas preventivas, como utilização de gemas livres de viroides aliadas a métodos confiáveis de indexação. A proposta desta revisão é apresentar ao leitor os recentes avanços nas pesquisas com viroides de citros, principalmente na taxonomia, distribuição geográfica, métodos de detecção, limpeza e indexação, epidemiologia e controle, além do histórico e importância desses patógenos para a citricultura mundial.Viroids are the smallest known plant pathogens consisting of a non-encapsidated, circular, single-stranded RNA that replicate autonomously in their host plants. Viroids are classified into two families (Pospiviroidae and Avsunviroidae). All citrus viroids belong to the Pospiviroidae family (species that present a central conserved region, replicate in the nucleus of infected cells and lack of ribozyme activity) with five citrus viroid species: Citrus exocortis viroid, CEVd (Pospiviroid), Hop stunt viroid, HSVd (Hostuviroid), Citrus bark cracking viroid, CBCVd (Cocadviroid) and Citrus bent leaf viroid, CBLVd and Citrus dwarfing viroid, CDVd (Apscaviroid). In addition, Citrus viroid original source (CVd-OS) and, more recently, Citrus viroid V (CVd-V) have been proposed as tentative species of the genus Apscaviroid. Citrus viroids are graft-transmitted and their dissemination occurs mainly by propagation of contaminated material. It is known that they have a broad host range, infecting species of citrus and plants of citrus-related genera. Two important diseases in citrus are viroid-induced: (i) exocortis; (ii) and cachexia. Symptoms vary from severe to asymptomatic, and their control is based on preventive measures as availability of viroid-free budwood as a source of propagation material follow by adequate indexing procedures. The purpose of this review is to present the reader the recent advances on citrus viroid research, mainly on taxonomy, geographical distribution, methods of detection, indexing and cleaning, epidemiology and control.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

GATA3 protein as a MUC1 transcriptional regulator in breast cancer cells

Introduction Recent studies have demonstrated that members of the GATA-binding protein (GATA) family (GATA4 and GATA5) might have pivotal roles in the transcriptional upregulation of mucin genes (MUC2, MUC3 and MUC4) in gastrointestinal epithelium. The zinc-finger GATA3 transcription factor has been reported to be involved in the growth control and differentiation of breast epithelial cells. In SAGE (serial analysis of gene expression) studies we observed an intriguing significant correlation between GATA3 and MUC1 mRNA expression in breast carcinomas. We therefore designed the present study to elucidate whether MUC1 expression is regulated by GATA3 in breast cancer cells. Methods: Promoter sequence analysis of the MUC1 gene identified six GATA cis consensus elements in the 5′ flanking region (GATA1, GATA3 and four GATA-like sequences). Chromatin immunoprecipitation and electrophoretic mobility-shift assays were employed to study the presence of a functional GATA3-binding site. GATA3 and MUC1 expression was analyzed in vitro with a GATA3 knockdown assay. Furthermore, expression of GATA3 and MUC1 genes was analyzed by realtime RT-PCR and immunohistochemistry on breast cancer-specific tissue microarrays. Results: We confirmed the presence of a functional GATA3-binding site on the MUC1 promoter region in the MCF7 cell line. We determined that GATA3 knockdown assays led to a decrease in MUC1 protein expression in MCF7 and T47D cells. In addition, we detected a statistically significant correlation in expression between GATA3 and MUC1 genes at the mRNA and protein levels both in normal breast epithelium and in breast carcinomas (p = 0.01). GATA3 expression was also highly associated with estrogen receptor and progesterone receptor status (p = 0.0001) and tumor grade (p = 0.004) in breast carcinomas. Conclusion: Our study provides evidence indicating that GATA3 is probably a mediator for the transcriptional upregulation of MUC1 expression in some breast cancers.Facultad de Ciencias Médica

GATA3 protein as a MUC1 transcriptional regulator in breast cancer cells

INTRODUCTION: Recent studies have demonstrated that members of the GATA-binding protein (GATA) family (GATA4 and GATA5) might have pivotal roles in the transcriptional upregulation of mucin genes (MUC2, MUC3 and MUC4) in gastrointestinal epithelium. The zinc-finger GATA3 transcription factor has been reported to be involved in the growth control and differentiation of breast epithelial cells. In SAGE (serial analysis of gene expression) studies we observed an intriguing significant correlation between GATA3 and MUC1 mRNA expression in breast carcinomas. We therefore designed the present study to elucidate whether MUC1 expression is regulated by GATA3 in breast cancer cells. METHODS: Promoter sequence analysis of the MUC1 gene identified six GATA cis consensus elements in the 5' flanking region (GATA1, GATA3 and four GATA-like sequences). Chromatin immunoprecipitation and electrophoretic mobility-shift assays were employed to study the presence of a functional GATA3-binding site. GATA3 and MUC1 expression was analyzed in vitro with a GATA3 knockdown assay. Furthermore, expression of GATA3 and MUC1 genes was analyzed by real-time RT-PCR and immunohistochemistry on breast cancer-specific tissue microarrays. RESULTS: We confirmed the presence of a functional GATA3-binding site on the MUC1 promoter region in the MCF7 cell line. We determined that GATA3 knockdown assays led to a decrease in MUC1 protein expression in MCF7 and T47D cells. In addition, we detected a statistically significant correlation in expression between GATA3 and MUC1 genes at the mRNA and protein levels both in normal breast epithelium and in breast carcinomas (p = 0.01). GATA3 expression was also highly associated with estrogen receptor and progesterone receptor status (p = 0.0001) and tumor grade (p = 0.004) in breast carcinomas. CONCLUSION: Our study provides evidence indicating that GATA3 is probably a mediator for the transcriptional upregulation of MUC1 expression in some breast cancers

Efeito do Cowpea aphid-borne mosaic virus sobre crescimento e variação quantitativa de fenois totais e flavonoides de Passiflora edulis Sims

Cowpea aphid-borne mosaic virus induz o endurecimento do pericarpo do fruto, nanismo, mosaico foliar e bolhas em plantas de maracujá. Os teores totais de fenois e flavonoides de folhas sadias e artificialmente infectadas foram quantificados pelo método de Folin-Ciocalteu e reação com cloreto de alumínio. Alturas de todas as plantas foram medidas no início e no final do experimento. As plantas infectadas apresentaram alturas 80% menores do que as plantas sadias. Não houve diferença estatística nos teores de fenois totais e de flavonoides entre tratamentos

RNAi-mediated silencing of Mediterranean fruit fly (Ceratitis capitata) endogenous genes using orally-supplied double-stranded RNAs produced in Escherichia coli

BACKGROUND: The Mediterranean fruit fly (medfly), Ceratitis capitata Wiedemann, is a major pest affecting fruit and vegetable production worldwide, whose control is mainly based on insecticides. Double-stranded RNA (dsRNA) able to down-regulate endogenous genes, thus affecting essential vital functions via RNA interference (RNAi) in pests and pathogens, is envisioned as a more specific and environmentally-friendly alternative to traditional insecticides. However, this strategy has not been explored in medfly yet. RESULTS: Here, we screened seven candidate target genes by injecting in adult medflies gene-specific dsRNA hairpins transcribed in vitro. Several genes were significantly down-regulated, resulting in increased insect mortality compared to flies treated with a control dsRNA targeting the green fluorescent protein (GFP) complementary DNA (cDNA). Three of the dsRNAs, homologous to the beta subunit of adenosine triphosphate (ATP) synthase (ATPsynbeta), a vacuolar ATPase (V-ATPase), and the ribosomal protein S13 (RPS13), were able to halve the probability of survival in only 48 h after injection. We then produced new versions of these three dsRNAs and that of the GFP control as circular molecules in Escherichia coli using a two-self-splicing-intron-based expression system and tested them as orally-delivered insecticidal compounds against medfly adults. We observed a significant down-regulation of V-ATPase and RPS13 messenger RNAs (mRNAs) (approximately 30% and 90%, respectively) compared with the control medflies after 3 days of treatment. No significant mortality was recorded in medflies, but egg laying and hatching reduction was achieved by silencing V-ATPase and RPS13. CONCLUSION: In sum, we report the potential of dsRNA molecules as oral insecticide in medfly

Métodos de RT-PCR e de hibridização Dot Blot para detecção universal de tospovirus

Transcriptase reverse - polymerase chain reaction (RT-PCR) and dot blot hybridization with digoxigenin-labeled probes were applied for the universal detection of Tospovirus species. The virus species tested were Tomato spotted wilt virus, Tomato chlorotic spot virus, Groundnut ringspot virus, Chrysanthemum stem necrosis virus, Impatiens necrotic spot virus, Zucchini lethal chlorosis virus, Iris yellow spot virus. Primers for PCR amplification were designed to match conserved regions of the tospovirus genome. RT-PCR using distinct primer combinations was unable to simultaneously amplify all tospovirus species and consistently failed to detect ZLCV and IYSV in total RNA extracts. However, all tospovirus species were detected by RT-PCR when viral RNA was used as template. RNA-specific PCR products were used as probes for dot hybridization. This assay with a M probe (directed to the G1/G2 gene) detected at low stringency conditions all Tospovirus species, except IYSV. At low stringency conditions, the L non-radioactive probe detected the seven Tospovirus species in a single assay. This method for broad spectrum detection can be potentially employed in quarantine services for indexing in vitro germplasm.Visando a um método para a detecção universal de tospovírus, utilizaram-se as técnicas de "Transcriptase reverse - polymerase chain reaction" (RT-PCR) e hibridização com sondas marcadas com digoxigenina. As espécies de tospovirus testadas foram: Tomato spotted wilt virus, Tomato chlorotic spot virus, Groundnut ringspot virus, Chrysanthemum stem necrosis virus, Impatiens necrotic spot virus, Zucchini lethal chlorosis virus, Iris yellow spot virus. Os oligonucleotídeos foram sintetizados para anelar em regiões conservadas do genoma viral, sendo os produtos de PCR utilizados como sondas para hibridização através de "dot blot". Através de RT-PCR, utilizando-se diferentes combinações de oligonucleotídeos, somente foi possível a amplificação de todas as espécies quando se utilizou RNA de vírus purificado, sendo que, para a detecção a partir de RNA total, a RT-PCR apresentou problemas para a detecção das espécies ZLCV e IYSV. Sob condições de baixa adstringência, os testes de hibridização por "dot blot" com a sonda M (correspondente ao gene G1/G2) detectaram todas as espécies testadas, exceto IYSV. Com a sonda L, também sob condições de baixa adstringência, pôde-se detectar todas as espécies de tospovirus testadas simultaneamente em um único ensaio. Este método de detecção pode ser utilizado em serviços de quarentena e para indexação de germoplasma in vitro

Produção de anti‑soros policlonais a partir de proteínas capsidiais recombinantes de Grapevine leafroll‑associated virus 2 e Grapevine virus B

The objective of this work was to produce and characterize specific antisera against Brazilian isolates of Grapevine leafroll-associated virus 2 (GLRaV-2) and Grapevine virus B (GVB), developed from expressed coat proteins (CPs) in Escherichia coli, and to test their possible use for the detection of these two viruses in diseased grapevines. The coat protein (CP) genes were RT-PCR-amplified, cloned and sequenced. The CP genes were subsequently subcloned, and the recombinant plasmids were used to transform E. coli cells and express the coat proteins. The recombinant coat proteins were purified, and their identities were confirmed by SDS-PAGE and Western blot and used for rabbit immunizations. Antisera raised against these proteins were able to recognize the corresponding recombinant proteins in Western blots and to detect GLRaV-2 and GVB in infected grapevine tissues, by indirect ELISA, discriminating healthy and infected grapevines with absorbances (A405) of 0.08/1.15 and 0.12/1.30, respectively. Expressing CP genes can yield high amount of viral protein with high antigenicity, and GLRaV-2 and GVB antisera obtained in this study can allow reliable virus disease diagnosis.O objetivo deste trabalho foi produzir e caracterizar anti-soros específicos contra isolados brasileiros do Vírus do enrolamento-da-folha da videira 2 (GLRaV-2) e do Vírus B da videira (GVB), desenvolvidos a partir das proteínas capsidiais expressas em Escherichia coli, e testar seu possível uso para a detecção destes dois vírus em videiras infectadas. Os genes da proteína capsidial (CP) foram amplificados via RT-PCR, clonados e seqüenciados. Foram, subseqüentemente, subclonados, e os plasmídeos recombinantes foram empregados na transformação das células de E. coli e na expressão das proteínas capsidiais. As proteínas capsidiais recombinantes foram purificadas, e suas identidades foram confirmadas em SDS-PAGE e "Western blot" e utilizadas para imunizar coelhos. Os anti-soros produzidos contra essas proteínas foram capazes de reconhecer as proteínas recombinantes correspondentes em "Western blot", de detectar GLRaV-2 e GVB em tecidos infectados de videiras pelo ELISA indireto, e de discriminar videiras sadias e infectadas, com absorbâncias (A405) de 0,08/1,15 e 0,12/1,30, respectivamente. A expressão dos genes CP pode produzir grandes quantidades de proteínas virais, com elevada antigenicidade, e os anti-soros de GLRaV-2 e GVB obtidos neste trabalho possibilitam a diagnose confiável desses vírus