An immunofluorescence test for diagnosis of ophthalmic herpes in a mouse corneal model

Herpes simplex virus type 1 (HSV-1) ophthalmic disease is the most common cause of corneal blindness in humans world-wide. Current culture techniques for HSV take several days and commercially available HSV laboratory based diagnostic techniques vary in sensitivity. Our study was conducted to evaluate the use of a quicker and simpler method to herpes ophthalmic diagnosis. Corneal smears were made by firm imprints of infected mouse eyes to glass slides, after smears were fixated with cold acetone, and an indirect immunofluorescence (IIF) method was performed using monoclonal antibodies in a murine model of ophthalmic herpes. Eye swabs from infected mice were inoculated in Vero cells for virus isolation. Cytology and histology of the eye were also performed, using hematoxylin-eosin routine. Mouse eyes were examined by slit-lamp biomicroscopy for evidence of herpetic disease at various times postinoculation. We made a comparative evaluation of sensitivity, specificity and speed of methods for laboratory detection of HSV. Our results indicate that this IIF method is quick, sensitive, specific and can be useful in the diagnosis of ophthalmic herpes as demonstrated in an animal model.

Deep Ecology: Princípios, Fundamentos e Fins

Dissertação apresentada para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Ecologia Humana e Problemas Sociais Contemporâneos

Simultaneous quantitation of serum HBV DNA and HBeAg can distinguish between slow and fast viral responses to antiviral therapy in patients with chronic hepatitis B

BACKGROUND: The quantitation of serum HBeAg is not commonly used to monitor viral response to therapy in chronic hepatitis B. METHODS: In this study, 21 patients receiving varying therapies were followed and their viral response monitored by concomitant viral load and HBeAg quantitation in order to study the meaning and the kinetics of both parameters. RESULTS: It was possible to distinguish between three different patterns of viral response. The first was characterized by a simultaneous decrease in serum HBV DNA and HBeAg. The second pattern was characterized by a decrease in serum HBeAg but persistent detection of HBV DNA. The third pattern was characterized by undetectable HBV DNA with persistent HBeAg positivity, which points to a non-response (Pattern III-B) except when HBeAg levels showed a slow but steady drop, characterizing a "slow responder" patient (Pattern III-A). CONCLUSIONS: The first pattern is compatible with a viral response. A long-term HBeAg seropositivity with a slow and persistent decrease (Pattern III-A) is also compatible with a viral response and calls for a prolongation of anti-viral treatment.

Rapid HIV diagnostic test in undocumented pregnant women applied at an inner-city teaching hospital

A significant number of Brazilian gestational-age women are still not tested for HIV, representing a high risk of transmission to their newborns. The current study sought to identify the number of pregnant women with no previous testing or undocumented for HIV referred to the Gynecology and Obstetrics Department of a Regional Teaching Hospital and included diagnosis of HIV infection determined by a rapid test and perinatal transmission in pregnancy. Medical records of all pregnant women admitted to hospital from January 2001 to December 2005 were reviewed. Pregnant women without HIV results were submitted to a rapid HIV test. Those who tested positive were further tested by ELISA and confirmed by indirect immunofluorescence assay (IIA) or Western blot (WB). The viral load from babies born to HIV-infected mothers was assessed by bDNA. Of the 16,424 pregnant women analyzed (6.6%), 1,089 were undocumented for HIV. Eleven women were positive in rapid testing and 10 were confirmed by ELISA, IIA or WB, with 0.9% seropositivity. Mother/infant pairs received zidovudine monotherapy prophylaxis and infant viral load was lower than 50 copies/mL. A higher number of pregnant women previously tested for HIV during antenatal care was verified, compared to that obtained nationwide.

Pseudo-outbreak of Clostridium difficile associated diarrhea (CDAD) in a tertiary-care hospital

The objective of this study was to describe a pseudo-outbreak of C. difficile in a hospital, following a change in the method used to detect the toxin. In February 2002, there were two cases of CDAD and in March 7 occurred, coinciding with a change of the test (from detection of toxin A to toxin A/B). An outbreak was suspected. Active surveillance and education of staff were started. A CDAD case was defined as a patient with acute onset of diarrhea (³ three episodes of liquid stools) and a positive stool test. They were classified as hospital or community-acquired. Stool samples were also collected for C. difficile culture and isolates were typed using AP-PCR. From March 2002 through December 2003 there were 138 cases of CDAD: 70% were hospital-acquired and among the 30% with CDAD present on admission, most (81%) came directly from the community (50% had no history of hospitalization). Fifty-two percent of hospital-acquired CDAD and 94% of cases on admission had already used antibiotics. The incidence of CDAD in hospitalized patients during surveillance was 3.3 per 1000 patient-admissions. The incidence of CDAD present on admission was 6.1/1000 patients. Sixteen isolates were typed and presented 13 different profiles. In conclusion, the CDAD increase in our study occurred due to change in diagnostic methods and not due to an outbreak, as suspected initially. The incidence in hospitalized patients was much lower than in reported outbreaks. There were 13 molecular types suggesting that an outbreak did not occur. CDAD was largely community-acquired.

On an acute case of Chagas disease in a region under vector control in the state of São Paulo, Brazil

No vector transmitted cases of Chagas disease had been notified in the state of São Paulo since the 1970s. However, in March, 2006, the death of a six-year-old boy from the municipality of Itaporanga was notified to the Center for Epidemiological Survey of the São Paulo State Health Secretariat: an autochthonous case of acute Chagas disease. The postmortem histopathological examination performed in the Hospital das Clínicas of the Botucatu School of Medicine confirmed the diagnosis. Reference to hospital records, consultation with the health professionals involved in the case and interviews with members of the patient's family supplied the basis for this study. We investigated parasite route of transmission, probable local reservoirs and vectors. No further human cases of acute Chagas disease were diagnosed. No locally captured vectors or reservoirs were found infected with Trypanosoma cruzi. Alternative transmission hypotheses - such as the possible ingestion of foods contaminated with vector excreta - are discussed, as well as the need to keep previously endemic regions and infested houses under close surveillance. Clinicians should give due attention to such signs as uni- or bilateral palpebral edema, cardiac failure, myocarditis, pericarditis, anasarca and atypical signs of nephrotic syndrome or nephritis and consider the diagnostic hypothesis of Chagas disease.

Effectiveness of the Brazilian influenza vaccination policy, a systematic review

Since 1999, Brazil has undertaken annual influenza vaccine campaigns, free of charge, targeting the elderly population, health professionals, and immune-deficient patients. We conducted a systematic review of literature in order to evaluate the effectiveness of the initiative. We used the keywords influenza, vaccine, Brazil and effectiveness to search the main databases. Thirty-one studies matched our inclusion and exclusion criteria. Influenza vaccine coverage among the elderly is high, though not as high as suggested by the official figures. Estimates on effectiveness are scarce. The majority come from ecological studies that show a modest reduction in mortality and hospital admissions due to influenza-related causes. Such reduction is not evident in the North and Northeastern states of Brazil, a finding that is probably related to the different seasonal pattern of influenza in equatorial and tropical regions. Brazilian epidemiologists still owe society better-designed studies addressing the effectiveness of influenza vaccine campaigns.

Evaluation of Helicobacter pylory colonization by serologic test (IgG) and dyspepsia in volunteers from the countryside of Monte Negro, in the Brazilian western Amazon region

The present study intended to analyze the seroprevalence of Helicobacter pylori, IgG, and its relation to dyspepsia in a population from the western Amazon region. During the "Projeto Bandeira Científica", a University of São Paulo Medical School program, in Monte Negro's rural areas, state of Rondônia, 266 blood samples were collected from volunteers. The material was tested for IgG antibodies anti-Helicobacter pylori by ELISA method and the participants were also interviewed on dyspepsia, hygiene and social aspects. Participants aged between five and 81 years old (34 years on average), 149 (56%) were female and 117 (44%) male. We found 210 (78.9%) positive, 50 (18.8%) negative and six (2.3%) undetermined samples. Dyspeptic complaints were found in 226 cases (85.2%). There was no statistical association between dyspepsia and positive serology for H. pylori. We concluded that the seroprevalence in all age categories is similar to results found in other studies conducted in developing countries, including those from Brazil. On the other hand, the seroprevalence found in Monte Negro was higher than that reported in developed countries. As expected, there was a progressive increase in the positivity for H. pylori in older age groups.

Intervalos melódicos: diferentes estratégias de aprendizagem

O presente documento foi realizado no âmbito do Mestrado em Ensino de Música, ramo Formação Musical, na Escola Superior de Educação e Escola Superior de Música Artes e Espectáculos do Instituto Politécnico do Porto. O objectivo central deste documento visa, num primeiro capítulo, contextualizar o meio, a escola e a turma. No segundo capítulo deste documento constará a reflexão da Prática Pedagógica em Formação Musical, no Conservatório de Música de Paredes. No terceiro capítulo será apresentado o Projecto de Investigação em estudo – Intervalos melódicos: diferentes estratégias de aprendizagem. Esta investigação surge no sentido de tentar perceber quais as estratégias que surtem efeitos mais positivos para a aprendizagem de intervalos melódicos. As estratégias adoptadas foram três, são elas: aprendizagem por canções; aprendizagem por grau conjunto; aprendizagem por canções e grau conjunto. Foram seleccionados 18 indivíduos com idades compreendidas entre os 8 e os 12 anos de idade, de ambos os sexos. Todos eles têm em comum o facto de frequentarem o 1º grau de Formação Musical. A metodologia utilizada foi o estudo de caso, com abordagem quantitativa. Para testar as diferentes estratégias de aprendizagem foi realizado um teste após um período de oito semanas (8 aulas). Será feita uma revisão bibliográfica, onde serão abordadas algumas perspectivas de diferentes autores, para que possamos compreender melhor o tema desta investigação. As conclusões deste estudo revelam que a aprendizagem de intervalos melódicos utilizando a complementaridade das estratégias por canções e por grau conjunto são mais positivas, ainda que os resultados não sejam muito evidentes.

Audição harmónica: importância e estratégias para um desenvolvimento auditivo integral

O presente relatório insere-se no âmbito da unidade curricular de Prática de Ensino Supervisionada. É um trabalho individual de síntese e de reflexão fundamentada sobre o percurso realizado no estágio, e de investigação com vista à resolução de problemas e conhecimento da realidade das práticas educativas do sistema de ensino especializado de música. Deste modo, é definido o quadro contextual sobre a evolução deste sistema de ensino e as complexidades que o têm caraterizado, o desenvolvimento curricular, particularmente o lugar que a disciplina de Formação Musical ocupa no currículo, e o Conservatório de Música de Paredes. Sobre o percurso formativo desempenhado nesta instituição educativa e sobre os dados recolhidos da observação naturalista não participante das aulas de duas turmas do ensino articulado, é elaborada uma reflexão crítica fundamentada, relativamente a todos os processos didáticos e pedagógicos. Complementarmente é apresentado um projeto de investigação qualitativa realizado no âmbito da unidade curricular de Seminário de Investigação em ensino de Música, tendo em vista a contribuição para uma prática pedagógica mais enriquecida, informada e fundamentada. O objeto de estudo está relacionado com a área disciplinar de Formação Musical, sob o tema Audição Harmónica: importância e estratégias para um desenvolvimento auditivo integral. Os objetivos do projeto visam verificar: i) quais os recursos utilizados e as estratégias e metodologias de ensino praticadas nas aulas de Formação Musical para o desenvolvimento da audição harmónica; ii) quais os objetivos e conteúdos contemplados nos programas da mesma disciplina; iii) que atividades são sugeridas em alguns manuais de Formação Musical. Os dados obtidos permitiram verificar que, apesar da importância atribuída à audição harmónica na formação de músicos, esta competência representa uma das maiores dificuldades dos alunos e, no âmbito da Formação Musical, há pouca diversidade de vivências, experiências musicais e outras atividades contextualizadas musicalmente.

HIV-1-mother-to-child transmission and associated characteristics in a public maternity unit in Presidente Prudente, Brazil

In children, vertical transmission is the main form of HIV infection. Our aim was to determine the prevalence of HIV-1 vertical transmission in mother-infant pairs in a public maternity ward in Presidente Prudente, SP. Additionally; we sought to identify characteristics associated with this form of transmission. The files of 86 HIV-1-infected mothers and their newborns referred to a Public Hospital from March 2002 to March 2007 were analyzed. The HIV-1-RNA viral load of the newborns was determined by bDNA. The HIV-1 vertical-transmission rate was 4.6%. Children that were born in the pre-term period and breastfed were at a higher risk of HIV-1 infection (p = 0.005 and p = 0.017 respectively) than children born at term and not breastfed. Prophylactic therapy with zidovudine after birth for newborns was associated with a lower risk of infection (p = 0.003). The number of newborns weighing < 2,500 g was significantly higher for infected children (p = 0.008) than for non-infected newborns. About 22.9% of mothers did not know the HIV-1 status of their newborns eight months after delivery. The study suggests that it is necessary to increase the identification of HIV-1 infection in pregnant women and their newborns as well as to offer and explain the benefits of ARV prophylaxis.

Diagnosing visceral leishmaniasis and HIV/AIDS co-infection: a case series study in Pernambuco, Brazil

HIV/AIDS-associated visceral leishmaniasis may display the characteristics of an aggressive disease or without specific symptoms at all, thus making diagnosis difficult. The present study describes the results of diagnostic tests applied to a series of suspected VL cases in HIV-infected/AIDS patients admitted in referral hospitals in Pernambuco, Brazil. From a total of 14 eligible patients with cytopenias and/or fever of an unknown etiology, and indication of bone marrow aspirate, 10 patients were selected for inclusion in the study. Diagnosis was confirmed by the following examinations: Leishmania detection in bone marrow aspirate, direct agglutination test, indirect immunofluorescence, rK39 dipstick test, polymerase chain reaction and latex agglutination test. Five out of the ten patients were diagnosed with co-infection. A positive direct agglutination test was recorded for all five co-infected patients, the Leishmania detection and latex agglutination tests were positive in four patients, the rK39 dipstick test in three, the indirect immunofluorescence in two and a positive polymerase chain reaction was recorded for one patient. This series of cases was the first to be conducted in Brazil using this set of tests in order to detect co-infection. However, no consensus has thus far been reached regarding the most appropriate examination for the screening and monitoring of this group of patients.

Otimização da operação de picking em armazéns

A Logística, vista como uma perspetiva integradora entre os parceiros de negócio, com objetivos comuns de proporcionar ao cliente mais-valias e aspetos diferenciadores perante os outros concorrentes, contribui em muito na manutenção das empresas na globalização atual, que se torna cada vez mais flexível. Através de uma boa gestão de processos críticos de negócio, boa localização dos materiais, sejam eles quais forem, produtos finais, matérias-primas ou produtos em vias de fabrico e através do transporte a logística cria utilidade temporal e diferenciadora. De facto, a logística poderá assumir um papel fundamental em proporcionar valor acrescentado ao disponibilizar, a tempo, os serviços que os clientes necessitam ou esperam. Enquadrando-se na temática de gestão dos armazéns, o presente projeto consistiu no estudo de operações de picking com a finalidade de otimização dos processos de picking no armazém do operador logístico AR – Serviços de Logística, localizado em Ribeirão, Vila Nova de Famalicão. O trabalho inicial passou pelo levantamento do funcionamento das operações do processo de picking na empresa e posteriormente confrontá-los com as tecnologias e procedimentos atuais no mercado. Com base nos resultados obtidos, foi possível definir e implementar métricas enquadradas nas finalidades estratégicas e operacionais do operador logístico. As soluções passaram também pela melhoria da aplicação de gestão de armazéns (WMS), reavaliação dos indicadores previamente estabelecidos e na aquisição de equipamentos para automatização das operações picking e localizações. Os registos e informações relacionadas com os módulos fulcrais são armazenados e tratados na base de dados de suporte à aplicação com contributo de melhoria contínua aos procedimentos logístico da empresa e sua relação com os stakeholders na estratégia global de negócio com o operador logístico. Finalmente, foi possível analisar os resultados obtidos em modo real em relação as estimativas calculadas e definidas na fase de implementação e desenvolvimento.

Genetic diversity and primary resistance among HIV-1-positive patients from Maringá, Paraná, Brazil

The objective of this study is to identify subtypes of Human Immunodeficiency Virus type 1 (HIV-1) and to analyze the presence of mutations associated to antiretroviral resistance in the protease (PR) and reverse transcriptase (RT) regions from 48 HIV-1 positive treatment naïve patients from an outpatient clinic in Maringá, Paraná, Brazil. Sequencing was conducted using PR, partial RT and group-specific antigen gene (gag) nested PCR products from retrotranscribed RNA. Transmitted resistance was determined according to the Surveillance Drug Resistance Mutation List (SDRM) algorithm. Phylogenetic and SimPlot analysis of concatenated genetic segments classified sequences as subtype B 19/48 (39.6%), subtype C 12/48 (25%), subtype F 4/48 (8.3%), with 13/48 (27.1%) recombinant forms. Most recombinant forms were B mosaics (B/F 12.5%, B/C 10.4%), with one C/F (2.1%) and one complex B/C/F mosaic (2.1%). Low levels of transmitted resistance were found in this study, 2/48 (2.1% to NRTIs and 2.1% for PI). This preliminary data may subsidize the monitoring of the HIV evolution in the region.

Os diferentes tipos de resultados nas cooperativas numa perspetiva jurídica e contabilística

As cooperativas assumem um papel fundamental na economia social portuguesa, o que justifica a necessidade de um conhecimento mais aprofundado destas entidades tanto a nível jurídico como a nível contabilístico. O principal objetivo deste estudo consiste em identificar os vários tipos de resultados nas cooperativas, compreender e caraterizar o tratamento jurídico e contabilístico dos mesmos e aferir se o normativo contabilístico em vigor permite evidenciar as especificidades daqueles. Assim, utilizando uma metodologia qualitativa, com recurso à análise de conteúdo, realizou-se um estudo de caso múltiplo. Antes, porém, procedeu-se à revisão de literatura, seletiva e seminal, sobre o estado da arte dos resultados das cooperativas em Portugal. Foram identificados três tipos de resultados, os resultados cooperativos, os resultados extracooperativos e os resultados extraordinários. O estudo mostra que o tratamento jurídico e contabilístico dos resultados cooperativos não está devidamente adequado à realidade das mesmas, por duas razões: em primeiro lugar, não são claramente identificáveis na legislação em vigor os diferentes tipos de resultados; e, em segundo lugar, constata-se que os resultados nas cooperativas têm o mesmo tratamento contabilístico que os resultados das sociedades comerciais, apesar das diferenças substanciais entre as duas formas jurídicas. Concluiu-se, então que a alteração do enquadramento contabilístico aplicável às cooperativas é pertinente e necessária no sentido de impor uma contabilização separada dos resultados cooperativos e dos resultados extracooperativos e extraordinários. Esta contabilização separada é essencial, por razões fiscais mas sobretudo para que as demonstrações financeiras apresentem a imagem verdadeira e apropriada do desempenho das cooperativas. Conclui-se, de igual modo, que existe no ordenamento português um normativo contabilístico aplicável às entidades do setor não lucrativo que seria adequado para evidenciar as especificidades das cooperativas em matéria dos resultados, dado que o modelo das demonstrações dele constante permitiria o desdobramento dos resultados por atividades.