Xylella fastidiosa gene expression analysis by DNA microarrays

Xylella fastidiosa genome sequencing has generated valuable data by identifying genes acting either on metabolic pathways or in associated pathogenicity and virulence. Based on available information on these genes, new strategies for studying their expression patterns, such as microarray technology, were employed. A total of 2,600 primer pairs were synthesized and then used to generate fragments using the PCR technique. The arrays were hybridized against cDNAs labeled during reverse transcription reactions and which were obtained from bacteria grown under two different conditions (liquid XDM2 and liquid BCYE). All data were statistically analyzed to verify which genes were differentially expressed. In addition to exploring conditions for X. fastidiosa genome-wide transcriptome analysis, the present work observed the differential expression of several classes of genes (energy, protein, amino acid and nucleotide metabolism, transport, degradation of substances, toxins and hypothetical proteins, among others). The understanding of expressed genes in these two different media will be useful in comprehending the metabolic characteristics of X. fastidiosa, and in evaluating how important certain genes are for the functioning and survival of these bacteria in plants.

Análise das relações taxonômicas e sistemáticas entre espécies de triatomíneos (Hemiptera, Reduviidae) de colônias mantidas pelo Serviço Especial de Saúde de Araraquara, inferida de seqüências do 16S rDNA mitocondrial

Foram analisadas seqüências de nucleotídeos do gene 16S do rDNA mitocondrial em 14 populações de triatomíneos mantidos em colônias no insetário SESA de Araraquara- SP, comparando-as com seqüências do mesmo gene disponíveis no GenBank. Os fragmentos variaram de 311 a 317 pb com baixa variação intra-específica entre as distâncias genéticas (0% a 0,6%), exceto para os relacionamentos entre espécimes de Triatoma sordida (1%) e espécimes de T. brasiliensis (1,3%) atribuídos a populações geográficas diferentes. A parafilia de Rhodniini e do gênero Panstrongylus foi evidenciada pelas analises, confirmando resultados anteriores entre estes e os estreitos relacionamentos de R. prolixus com R. robustus e de T. infestans e T. platensis. O relacionamento entre T. maculata e T. pseudomaculata não foi solucionado, uma vez que, esses táxons apareceram tanto em monofilia quanto em parafilia: T. pseudomaculata (SESA) está agrupado com T. maculata (seqüência do GenBank) e associados a T . brasiliensis (SESA), enquanto T. maculata (SESA) aparece agrupado com T. pseudomaculata do SESA e do GenBank. Os resultados evidenciam a utilidade do gene 16S como marcador de espécies de triatomíneos e sua importância em questões de sistemática e taxonomia. Há necessidade de novos estudos envolvendo outros marcadores associados a caracteres sistemáticos clássicos de morfologia, ecologia e comportamento para decisões sistemáticas adequadas uma vez, que teriam impacto não apenas sistemático mas, para as estratégias de controle.

A regionalização intramunicipal do Sistema Único de Saúde (SUS): um estudo de caso do município de São Paulo-SP, Brasil

A regionalização tem sido apontada como um dos principais desafios para viabilizar a equidade e a integralidade do SUS. Este artigo tem como objetivo avaliar o processo de implementação de um projeto de organização de regiões de saúde no município de São Paulo. Para tanto, foi realizado um estudo de caso em uma região selecionada desse município, a partir do referencial da análise de implantação, utilizando-se como fonte de dados documentos da gestão e entrevistas semiestruturadas com informantes-chave da gestão municipal 2005-2008. A análise temática evidenciou que o projeto de regionalização idealizado no início da gestão não foi efetivamente implementado. Dentre os fatores que interferiram nesse insucesso, destacam-se: a) a Secretaria Municipal de Saúde (SMS), além de seu caráter centralizador, manteve estruturas político-administrativas independentes para a gestão da atenção básica e da assistência hospitalar; b) a SMS não assumiu a gestão, de fato, de ambulatórios e hospitais estaduais; c) o poder institucional e a resistência dos hospitais em se integrar ao sistema de saúde. Discute-se, ainda, a necessidade de avançar na descentralização intramunicipal do SUS e buscar novas estratégias para a construção de pactos que consigam superar as resistências e articular instituições historicamente consolidadas, visando uma regionalização cooperativa e solidária.

Vascular density and distribution of vascular endothelial growth factor (VEGF) and its receptor VEGFR-2 (Flk-1) are significantly higher in patients with deeply infiltrating endometriosis affecting the rectum

Objective: To analyze vascular density and immunolocalization of angiogenic vascular endothelial growth factor (VEGF) and its receptor Flk-1 in the proliferative and secretory eutopic human endometrium. and in three different sites of endometriosis: the ovary, bladder, and rectum. Design: Prospective study. Setting: University hospital. Patient(s): Thirty women with endometriosis (10 ovarian, 1.0 bladder, 10 rectal) and 32 control women (10 proliferative endometrium, 10 secretory endometrium, 4 normal ovary, 4 normal bladder, 4 normal rectum). Intervention(s): Normal endometrial samples were obtained from women during laparoscopic ablation of subserous myoma, and biopsy specimens of endometriosis were obtained from patients undergoing surgery for the diagnosis and treatment of endometriosis. Normal tissues of ovary, bladder, and rectum were obtained from these organs beside the lesions of endometriosis. Main Outcome Measure(S): Blood vessels were quantified according to the number of von Willebrand factor-positive endothelial cells. The VEGF and Flk-1 distribution were evaluated semiquantitatively by immunohistochemical staining. Result(s): More blood vessels were found in cases of endometriosis, particularly rectal endometriosis, compared with the respective control samples and with the eutopic endometrium, and they were localized in endometrial stroma around the glands. The VEGF and Flk-1 expression levels were also higher in cases of endometriosis, especially rectal endometriosis. Conclusion(s): Vascularization and VEGF and Flk-1 expression are significantly higher in deeply infiltrating endometriosis affecting the rectum, reinforcing the hypothesis that antiangiogenesis therapy may constitute a new modality of treatment, especially in cases of deep endometriosis involving the rectum.

HSP65 DNA as therapeutic strategy to treat experimental paracoccidioidomycosis

The conventional treatment for paracoccidioidomycosis, the most prevalent mycosis in Latin America, involves long periods of therapy resulting in sequels and high frequency of relapses. The search for new alternatives of treatment is necessary. Previously, we have demonstrated that the hsp65 gene from Mycobacterium leprae shows prophylactic effects against murine paracoccidioidomycosis. Here, we tested the DNAhsp65 immunotherapy in BALB/c mice infected with Paracoccidioides brasiliensis, the agent of paracoccidioidomycosis. We observed an increase of Th1 cytokines accompanied by a reduction in fungal burden and pulmonary injury. These results provide new prospects for immunotherapy of paracoccidioidomycosis and other mycoses. (C) 2009 Elsevier Ltd. All rights reserved.

Blockade of the chemokine receptor CXCR2 ameliorates adjuvant-induced arthritis in rats

Background and purpose: Chemokine receptors CXCR1 and CXCR2 may mediate influx of neutrophils in models of acute and chronic inflammation. The potential benefits of oral administration of a CXCR1/2 inhibitor, DF 2162, in adjuvant-induced polyarthritis (AIA) were investigated. Experimental approach: A model of AIA in rats was used to compare the therapeutic effects of the treatment with DF2162, anti-TNF or anti-CINC-1 antibodies on joint inflammation and local production of cytokines and chemokines. Key results: DF2162 prevented chemotaxis of rat and human neutrophils induced by chemokines acting on CXCR1/2. DF2162 was orally bioavailable and metabolized to two major metabolites. Only metabolite 1 retained CXCR1/2 blocking activity. Treatment with DF2162 ( 15 mg kg(-1), twice daily) or metabolite 1, but not metabolite 2, starting on day 10 after arthritis induction diminished histological score, the increase in paw volume, neutrophil influx and local production of TNF, IL-1 beta, CCL2 and CCL5. The effects of DF2162 were similar to those of anti-TNF, and more effective than those of anti-CINC-1, antibodies. DF2162 prevented disease progression even when started 13 days after arthritis induction. Conclusions and implications: DF 2162, a novel orally-active non-competitive allosteric inhibitor of CXCR1 and CXCR2, significantly ameliorates AIA in rats, an effect quantitatively and qualitatively similar to those of anti-TNF antibody treatment. These findings highlight the contribution of CXCR2 in the pathophysiology of AIA and suggest that blockade of CXCR1/2 may be a valid therapeutic target for further studies aiming at the development of new drugs for treatment of rheumatoid arthritis.

Pediatric glioblastoma cell line shows different patterns of expression of transmembrane ABC transporters after in vitro exposure to vinblastine

Resistance to drug is a major cause of treatment failure in pediatric brain cancer. The multidrug resistance (MDR) phenotype can be mediated by the superfamily of adenosine triphosphate-binding cassette (ABC) transporters. The dynamics of expression of the MDR genes after exposure to chemotherapy, especially the comparison between pediatric brain tumors of different histology, is poorly described. To compare the expression profiles of the multidrug resistance genes ABCB1, ABCC1, and ABCG2 in different neuroepithelial pediatric brain tumor cell lines prior and following short-term culture with vinblastine. Immortalized lineages from pilocytic astrocytoma (R286), anaplasic astrocytoma (UW467), glioblastoma (SF188), and medulloblastoma (UW3) were exposed to vinblastine sulphate at different schedules (10 and 60 nM for 24 and 72 h). Relative amounts of mRNA expression were analyzed by real-time quantitative polymerase chain reaction. Protein expression was assessed by immunohistochemistry for ABCB1, ABCC1, and ABCG2. mRNA expression of ABCB1 increased together with augmenting concentration and time of exposure to vinblastine for R286, UW467, and UW3 cell lines. Interestingly, ABCB1 levels of expression diminished in SF188. Following chemotherapy, mRNA expression of ABCC1 decreased in all cell lines other than glioblastoma. ABCG2 expression was influenced by vinblastine only for UW3. The mRNA levels showed consistent association to protein expression in the selected sets of cell lines analyzed. The pediatric glioblastoma cell line SF188 shows different pattern of expression of multidrug resistance genes when exposed to vinblastine. These preliminary findings may be useful in determining novel strategies of treatment for neuroepithelial pediatric brain tumors.

Central effects of Tityus serrulatus and Tityus bahiensis scorpion venoms after intraperitoneal injection in rats

A great number of studies on scorpion venoms associate their effects to the autonomic nervous system, and few data are available about their action on the central nervous system (CNS). The aim of this work was to evaluate some central effects after intraperitoneal injection of Tityus serrulatus or T. bahiensis scorpion venoms. The hippocampal concentration of some neurotransmitters and their metabolites were determined. Electroencephalographic and behavioral observations were performed, and all brains were removed for histopathological analysis of hippocampal areas. Both venoms induced electrographic and behavioral alterations despite T bahiensis venom affects less the electrographic activity than T. serrulatus venom. Neurochemical analysis demonstrated no alteration in the extracellular levels of almost all the neurotransmitters evaluated, at least in the hippocampus, and no neuronal loss in this area was observed. Meanwhile, extracellular concentration of HVA increased up to 10 times in approximately 1/3 of the animals of both groups. Scorpion venoms seem to exert a small but important central effect. More studies in this field are necessary because they may be useful in developing new strategies to reduce the damage caused by scorpion stings. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

The endgame for national girls' schooling policies in Australia?

Gender equity: A framework for Australian schools is the most recent policy dealing with gender in schools at the national level in Australia. This paper provides a critical discourse analysis of the policy document, tracking two themes: 'the construction of gender', and 'equity: a discourse of education for all boys and girls'. Through this analysis the authors argue that the policy signals a substantial shift in focus-from girls and boys in relation to girls, to both girls and boys-within a framework of presumptive equality. It is also argued that the policy shuts down federal involvement in policy for girls' schooling. In the process, responsibility is devolved to the states and territories where, in many cases, gender equity programs will be struggled over at a local, school-based level. At the same time, however, spaces have been created which potentially enable new strategies for gender equity policies in Australian schools.

Novel miniaturized systems in high-throughput screening

High-throughput screening (HTS) using high-density microplates is the primary method for the discovery of novel lead candidate molecules. However, new strategies that eschew 2D microplate technology, including technologies that enable mass screening of targets against large combinatorial libraries, have the potential to greatly increase throughput and decrease unit cost. This review presents an overview of state-of-the-art microplate-based HTS technology and includes a discussion of emerging miniaturized systems for HTS. We focus on new methods of encoding combinatorial libraries that promise throughputs of as many as 100 000 compounds per second.

O aluno com deficiência motora e a acessibilidade arquitectónica no Ensino Básico : um estudo de caso : o concelho de Guimarães

RESUMO:Para que o sucesso educativo se verifique, a escola tem de ajustar a sua prática educativa a todos os alunos, incluindo os que apresentam necessidades educativas especiais (NEE). Portugal dispõe de legislação que assegura o direito à educação e à igualdade de oportunidades, consignados na Constituição da República Portuguesa (artigos 71º, 73º e 74º), em consonância com a Declaração Universal dos Direitos do Homem e na Lei de Bases do Sistema Educativo (artigos 2º, 7º, 17º e 18º). O nosso país subscreveu também a Declaração de Salamanca, a qual reuniu, em 1994, o consenso de noventa e dois governos e de vinte e cinco organizações internacionais, reafirmando o direito à educação para todos. A construção da escola inclusiva passa pela responsabilização da escola por todos os alunos, na perspectiva de educação para todos, exigindo a sua concretização novas estratégias e resposta a novos desafios. Há que alterar práticas para atender à diversidade. Assim, as escolas precisam de estar arquitectonicamente adaptadas às necessidades dos alunos que utilizam a cadeira de rodas como forma de mobilidade, respeitando o direito à diferença e possibilitando, em igualdade de oportunidades, o sucesso escolar e educativo. Levar à prática as medidas previstas na lei portuguesa e implementar, em cada escola, a inclusão do aluno com mobilidade condicionada, implica equipar as escolas com adaptações, meios e recursos educativos facilitadores do seu processo ensino/aprendizagem. A constatação do que efectivamente acontece no terreno educativo no que concerne aos alunos com deficiência motora que usam a cadeira de rodas para se movimentarem assume toda a centralidade da nossa investigação, que tem o cariz de um estudo de caso. Incide sobre a compreensão mais aprofundada de como se processa o acesso arquitectónico nas escolas do Ensino Básico da Zona Norte e mais concretamente do Concelho de Guimarães. Recolhemos dados através de questionários dirigidos aos professores de apoio educativo e coordenadores de escola; ouvimos os alunos com deficiência motora pronunciar-se sobre as dificuldades que sentiam; observámos o seu quotidiano escolar e, finalmente, escutámos as entidades locais, através de uma entrevista realizada à Vereadora da Cultura do Concelho de Guimarães. Os resultados obtidos serão, como foi acordado, divulgados aos intervenientes com responsabilidades directas no campo da educação que se prontificaram a partilhar connosco a sua informação e saber, no sentido de os sensibilizar para a necessidade de rever o aspecto das acessibilidades arquitectónicas nas instituições educativas que servem o seu concelho, pois, como verificámos, muito há ainda a fazer para que o Ensino Básico, universal e obrigatório, se traduza numa igualdade de oportunidades de acesso e sucesso educativo para os alunos que utilizam a cadeira de rodas para se movimentarem no espaço escolar. ABSTRACT: For the educational success of a school it has to adjust its educational practice so that it includes those with special needs. According to the Portuguese law, disabled people have the right to education and equal opportunities, well expressed in the Constitution of the Portuguese Republic (articles 71st, 73rd, and 74th), in consonance with the Universal Declaration of Human Rights, and in the main law the educational system (articles 2nd, 7th, 17th and 18th). Our country has also subscribed the Declaration of Salamanca, which was written in 1994 with the agreement of Representatives from ninety-two governments and twenty-five International Organizations, where was reaffirmed the right of education for all. It is necessary that school increases its responsibility in what concerns the education of all its students. Educating all and every student demands new strategies to face new challenges and so some practices have to be changed to respond to diversity. Schools must be architecturally adapted to give the disabled students, who use a wheel chair, equal opportunities to achieve educational success. Taking the measures foreseen in Portuguese law into practice, and implementing in each school the inclusion of students with conditioned mobility, involves equipping schools with adaptations, equipment and educational resources that facilitate the teaching / learning process. The observation of what actually happens on educative ground, regarding to pupils with motor disabilities who use a wheelchair to move, assumes the centrality of our research, which is a case study. It focuses on the deeper understanding of the way the architectonic access in primary schools of the North, and more specifically the Municipality of Guimarães, is processed. We collected data through questionnaires addressed to educational support teachers and school coordinators; heard students with motor disabilities to comment on the difficulties they felt; observed their daily school life, and finally heard the local authorities, through an interview the Councillor of Culture of the Municipality of Guimarães. As agreed, the results will be communicated to intervenients with direct responsibilities in the field of education who were willing to share with us their information and knowledge in order to raise awareness of the need of looking over the aspect of architectural accessibility in educational institutions that serve their county, because, as noted, much remains to be done so that the basic education, universal and compulsory, will result in equality of educational opportunities in access and success for students who use a wheelchair to move in school space.

The convergence of the Central American countries to International Accounting Standards

Abstract: In the last few decades, Central American countries are making a significant effort in order to modernize their governments' legislation both on financial management and systems of financial information. In this sense, these countries aim to enhance the quality of public financial information in order to improve decision-making processes, decrease the level of corruption, and keep citizens informed. In this context, the purpose of this paper is twofold. Firstly, to assess the degree of similarity of the financial information that is being developed by Central American governments with regard to the recommendations set up by Ipsas, and secondly, to analyse the efforts and the strategies that those countries are carrying out in the process of implementing those standards. To determine the differences in the information containing the annual financial statements issued by national public authorities and the recommendations set up by Ipsas we conducted a deductive content analysis. In view of the results we can say that the quality of annual financial statements presented by the countries in Central America, in comparison to the recommendations by the Ipsas concerning Ifac information, is not enough. Hence, in order to operate significant changes, it is still necessary to create new strategies for the implementation of the Ipsas.

How to teach mathematics with MOODLE

The aim of this article is to present a Project in the Oporto’s Institute of Accounting and Administration, which pretends to contribute for a change in the way of teaching and learning Mathematics. One of the main objectives of this project is to innovate the teaching and learning processes, exploring technologies as a pedagogical resource and to induce higher motivation to students, improve the rate of success and make available to students a set of materials adapted to their needs. This concern is justified due to the fact that students have a weak preparation, without consolidated basis. Since the year 2007/2008 the courses were adjusted to the Bologna process, which requires several changes in teacher’s and student’s roles, methodologies and assessment. The number of weekly classes has been reduced, so it was necessary to develop new strategies and methodologies to support the student. With the implementation of the Bologna Process in the Accounting degree, we felt a great need to provide other types of activities to students. To complement our theoretical and practical classes we have developed a project called MatActiva based on the Moodle platform offered by PAOL - Projecto de Apoio On-Line (Online Support Project). Moodle allows us to use the language TEX to create materials that use mathematical symbols. Using this functionality, we created a set of easy to use interactive resources. In MatActiva project, the students have access to a variety of different materials. We have followed a strategy that makes the project compatible with the theoretical and practical subjects/classes, complementing them. To do so, we created some resources, for instance multiple-choice tests, which are the most accessed by the students. These tests can be realized and corrected on-line and for each wrong answer there is a feedback with the resolution. We can find other types of resources: diagnostic tests, theoretical notes. There are not only the pre-requirements for subjects mathematics, but also materials to help students follow up the programs. We also developed several lessons. This activity consists of a number of pages, where each page has contents and leads to other pages, based on the student's progress. The teacher creates the choices and determines the next page that the student will see, based upon their knowledge. There is also an area of doubts, where the students can place all the mathematical doubts they have, and a teacher gives the answers or clues to help them in their work. MatActiva also offers an area where we can find some humour, curiosities, contests and games including mathematical contents to test the math skills, as well as links to pages about mathematical contents that could be useful for the study. Since ISCAP receives ERASMUS students and some of them attend mathematics, we developed some materials in English, so they can also use MatActiva. The main objectives of our project are not only to bring success in the subjects of mathematics, but also to motivate the students, encourage them to overcome theirs difficulties through an auto-study giving them more confidence and improve their relationship with the mathematics as well as the communication between students and teachers and among students.

A web-based intelligent tutoring system on teaching and learning electrical project

The Information and Communication Technology (ICT) provide new strategies for disseminating information and new communication models in order to change attitudes and human behaviour concerning to education. Nowadays the internet is crucial as a means of communication and information sharing. To education or tutorship will be required to use ICT, supported on the internet, to establish the communication of teacher-student and student-student, disseminating the content of the subjects, and as a way of teaching and learning process. This paper presents an intelligent tutor that aims to be a tool to support teaching and learning in the field of the electrical engineering project.

Barriers for HIV testing during pregnancy in Southern Brazil

OBJECTIVE: To assess HIV testing rate and determine risk factors for not have been tested during pregnancy. METHODS: A cross-sectional study was carried out in Porto Alegre, Southern Brazil, from December 2000 to February 2001. Socioeconomic, maternal and healthcare variables were obtained by means of a standardized questionnaire. Crude and adjusted odds ratios and their 95% confidence intervals were obtained in logistic regression models. RESULTS: A total of 1,642 mothers were interviewed. Of them, 94.3% reported being offered HIV testing before or during pregnancy or during labor; 89 mothers (5.4%) were not tested or did not know if they were tested. Attending fewer than six prenatal visits, being single and younger than 18 years old were relevant barriers preventing HIV testing. There was found a relationship between maternal schooling and the category of prenatal care provider. Having low 22.20 (12.43-39.67) or high 3.38 (1.86-7.68). schooling and being cared in the private sector strongly reduced the likelihood of being HIV tested. CONCLUSIONS: The Brazilian Health Ministry's recommendation for universal counseling and HIV testing has been successfully implemented in the public sector. In order to improve HIV testing coverage, new strategies need to target women cared in the private sector especially those of low schooling.