T CD4+ cells count among patients co-infected with human immunodeficiency virus type 1 (HIV-1) and human T-cell leukemia virus type 1 (HTLV-1): high prevalence of tropical spastic paraparesis/HTLV-1-associated myelopathy (TSP/HAM)

INTRODUCTION: HIV positive patients co-infected with HTLV-1 may have an increase in their T CD4+ cell counts, thus rendering this parameter useless as an AIDS-defining event. OBJECTIVE: To study the effects induced by the co-infection of HIV-1 and HTLV-1 upon CD4+ cells. MATERIAL AND METHODS: Since 1997, our group has been following a cohort of HTLV-1-infected patients, in order to study the interaction of HTLV-1 with HIV and/or with hepatitis C virus (HCV), as well as HTLV-1-only infected asymptomatic carriers and those with tropical spastic paraparesis/HTLV-1 associated myelopathy (TSP/HAM). One hundred and fifty HTLV-1-infected subjects have been referred to our clinic at the Institute of Infectious Diseases "Emílio Ribas", São Paulo. Twenty-seven of them were also infected with HIV-1 and HTLV-1-infection using two ELISAs and confirmed and typed by Western Blot (WB) or polymerase chain reaction (PCR). All subjects were evaluated by two neurologists, blinded to the patient's HTLV status, and the TSP/HAM diagnostic was based on the World Health Organization (WHO) classification. AIDS-defining events were in accordance with the Centers for Disease Control (CDC) classification of 1988. The first T CD4+ cells count available before starting anti-retroviral therapy are shown compared to the HIV-1-infected subjects at the moment of AIDS defining event. RESULTS: A total of 27 HIV-1/HTLV-1 co-infected subjects were identified in this cohort; 15 already had AIDS and 12 remained free of AIDS. The median of T CD4+ cell counts was 189 (98-688) cells/mm³ and 89 (53-196) cells/mm³ for co-infected subjects who had an AIDS-defining event, and HIV-only infected individuals, respectively (p = 0.036). Eight of 27 co-infected subjects (30%) were diagnosed as having a TSP/HAM simile diagnosis, and three of them had opportunistic infections but high T CD4+ cell counts at the time of their AIDS- defining event. DISCUSSION: Our results indicate that higher T CD4+ cells count among HIV-1/HTLV-1-coinfected subjects was found in 12% of the patients who presented an AIDS-defining event. These subjects also showed a TSP/HAM simile picture when it was the first manifestation of disease; this incidence is 20 times higher than that for HTLV-1-only infected subjects in endemic areas.

Optimization of the Sybr Green real time PCR for the detection of Human Herpes Virus type 6 (HHV-6)

HHV-6 is the etiological agent of Exanthem subitum which is considered the sixth most frequent disease in infancy. In immuno-compromised hosts, reactivation of latent HHV-6 infection may cause severe acute disease. We developed a Sybr Green Real Time PCR for HHV-6 and compared the results with nested conventional PCR. A 214 pb PCR derived fragment was cloned using pGEM-T easy from Promega system. Subsequently, serial dilutions were made in a pool of negative leucocytes from 10-6 ng/µL (equivalent to 2465.8 molecules/µL) to 10-9 (equivalent to 2.46 molecules/µL). Dilutions of the plasmid were amplified by Sybr Green Real Time PCR, using primers HHV3 (5' TTG TGC GGG TCC GTT CCC ATC ATA 3)'and HHV4 (5' TCG GGA TAG AAA AAC CTA ATC CCT 3') and by conventional nested PCR using primers HHV1 (outer): 5'CAA TGC TTT TCT AGC CGC CTC TTC 3'; HHV2 (outer): 5' ACA TCT ATA ATT TTA GAC GAT CCC 3'; HHV3 (inner) and HHV4 (inner) 3'. The detection threshold was determined by plasmid serial dilutions. Threshold for Sybr Green real time PCR was 24.6 molecules/µL and for the nested PCR was 2.46 molecules/µL. We chose the Real Time PCR for diagnosing and quantifying HHV-6 DNA from samples using the new Sybr Green chemistry due to its sensitivity and lower risk of contamination.

Metodologia BIM, aplicada à preparação, controlo e gestão de obra

O âmbito deste trabalho envolve o teste do modelo BIM numa obra em construção pela Mota-Engil – Engenharia, na extração experimental de peças desenhadas de preparação e apoio à execução de obra. No capítulo 1 deste relatório são definidos o âmbito e os objetivos deste trabalho, é feito um enquadramento histórico do tema e abordados conceitos e atividades da preparação de obra, na sua forma tradicional. O estado do conhecimento da preparação de obras e mais em concreto da tecnologia BIM a nível nacional e internacional é abordado no capítulo 2. Nesse sentido procura-se definir os conceitos principais inerentes a esta nova metodologia, que passa por identificar e caraterizar a tecnologia envolvida e o seu nível de desenvolvimento. Com suporte em casos práticos de preparação de obra na sua forma tradicional, identificados e desenvolvidos no capítulo 3, foi compilado um processo tipo de peças desenhadas de suporte identificadas e caracterizadas no capítulo 4, frequentes e comuns à execução de diversos tipos de obras de edifícios. Assente na compilação baseada em casos práticos e no estudo do projeto de execução da empreitada que sustenta o presente trabalho, com base no qual o modelo BIM foi concebido, identificou-se um conjunto de peças desenhadas de preparação e apoio à execução dos trabalhos, em 2D, a extrair do modelo. No capítulo 5, é feita uma descrição do modo como foi estudado o projeto da obra, com evidência para os fatores mais relevantes, especificando os desenhos a extrair. Suportada pelo programa de modelação ArchiCAD, a extração do conjunto de desenhos identificados anteriormente foi conseguida com recurso às funcionalidades disponíveis no software, que permite a criação de desenhos 2D atualizáveis ou não automaticamente a partir do modelo. Qualquer alteração introduzida no modelo virtual é automaticamente atualizada nos desenhos bidimensionais, caso o utilizador assim o pretenda. Ao longo desse trabalho foram detetados e analisados os condicionalismos inerentes ao processo de extração, referidos no capítulo 6, para estabelecimento de regras de modelação padrão a adotar em futuras empreitadas, que possam simplificar a obtenção dos elementos desenhados de preparação necessários à sua execução. No ponto 6.3 são identificadas melhorias a introduzir no modelo. Em conclusão no capítulo 7 são abordadas especificidades do setor da construção que sustentam e evidenciam cada vez mais a necessidade de utilizar as novas tecnologias com vista à adoção de práticas e ferramentas padrão de apoio à execução de obras. Sendo a tecnologia BIM, transversal a todo o setor, a sua utilização com regras padrão na conceção dos modelos e na extração de dados, potencia a otimização dos custos, do tempo, dos recursos e da qualidade final de um empreendimento, ao longo de todo o seu ciclo de vida, para além de apoiar com elevada fiabilidade as tomadas de decisão ao longo desse período. A tecnologia BIM, possibilita a antevisão do edifício a construir com um elevado grau de pormenor, com todas as vantagens que daí advêm.

Tensile fracture characterization of adhesive joints by standard and optical techniques

The use of adhesive joints has increased in recent decades due to its competitive features compared with traditional methods. This work aims to estimate the tensile critical strain energy release rate (GIC) of adhesive joints by the Double-Cantilever Beam (DCB) test. The J-integral is used since it enables obtaining the tensile Cohesive Zone Model (CZM) law. An optical measuring method was developed for assessing the crack tip opening (δn) and adherends rotation (θo). The proposed CZM laws were best approximated by a triangular shape for the brittle adhesive and a trapezoidal shape for the two ductile adhesives.

Molecular characterization of environmental Cryptococcus neoformans isolated in Vitoria, ES, Brazil

Cryptococcus neoformans is the major cause of fungal meningitis, a potentially lethal mycosis. Bird excreta can be considered a significant environmental reservoir of this species in urban areas, thirty-three samples of pigeon excreta were collected within the city of Vitoria, Brazil. Cryptococcus neoformans was isolated and identified using standard biochemical assays in ten samples. PCR amplification with primer M13 and orotidine monophosphate pyrophosphorylase (URA5) gene-restriction fragment length polymorphism (RFLP) analysis discerned serotypes and genotypes within this species. All isolates were serotype A (C. neoformans var. grubii) and genotype VNI. The two alternative alleles a and α at the mating type locus were determined by PCR amplification and mating assays performed on V8 medium. All isolates were MAT α mating type but only 50% were able to mate in vitro with the opposite mating type MAT a tester strains (JEC20, KN99a and Bt63). This study adds information on the ecology and molecular characterization of C. neoformans in the Southeast region of Brazil.

The relationship between knowledge acquisition on international assignments and career development: an exploratory study with portuguese repatriates

This paper explores the relationship between the expatriates’ knowledge acquisition (KA) and their career development after an international assignment (IA). The purpose of this paper is to examine the role of expatriates in KA and transfer within International Portuguese multinational corporations. Furthermore, with this empirical study we try to analyse how the knowledge that is acquired and transferred translates into a basis for career development after the IA. This phenomenon has a special relevance in the Portuguese context, because this country is known a growing process of globalization in recent years. Furthermore, (a) there are no empirical studies concerning knowledge transfer and career development of repatriates from Portuguese companies; (b) little is known about the repatriates’ contributions to their home company after IA. This paper is one of the first to focus specifically on the repatriates’ role in KA and transfer from the host company to their Portuguese home company. A qualitative research methodology is used, specifically through an exploratory case study approach, which examines how knowledge management (KM) acquisition or transferring during IA are important for the repatriates’ career development in the Portuguese home company. Data were collected through semi-structured interviews to 42 Portuguese international assignees and 18 organizational representatives from nine Portuguese companies. Preliminary results show that KA and transfer made by Portuguese expatriates contributes directly to their career development. Moreover, evidence reveals that not all repatriates were promoted after their IA; rather some repatriates were even demoted after their IA. Furthermore, the results obtained suggest that the type of knowledge which acquired or transferred plays a central role in the career development after repatriation. According to these results, the paper discusses the major theoretical and practical implications. Suggestions for future research are also presented.

Influence of human t-cell lymphotropic virus type 1 (HTLV-1) Infection on laboratory parameters of patients with chronic hepatitis C virus

Hepatitis C virus (HCV) and human T-cell lymphotropic virus type 1 (HTLV-1) share routes of transmission and some individuals have dual infection. Although some studies point to a worse prognosis of hepatitis C virus in patients co-infected with HTLV-1, the interaction between these two infections is poorly understood. This study evaluated the influence of HTLV-1 infection on laboratory parameters in chronic HCV patients. Twelve HTLV-1/HCV-coinfected patients were compared to 23 patients infected only with HCV, in regard to demographic data, risk factors for viral acquisition, HCV genotype, presence of cirrhosis, T CD4+ and CD8+ cell counts and liver function tests. There was no difference in regard to age, gender, alcohol consumption, smoking habits, HCV genotype or presence of cirrhosis between the groups. Intravenous drug use was the most common risk factor among individuals co-infected with HTLV-1. These patients showed higher TCD8+ counts (p = 0.0159) and significantly lower median values of AST and ALT (p = 0.0437 and 0.0159, respectively). In conclusion, we have shown that HCV/HTLV-1 co-infected patients differs in laboratorial parameters involving both liver and immunological patterns. The meaning of these interactions in the natural history of these infections is a matter that deserves further studies.

Renal Pathology in Portuguese HIV-Infected Patients

HIV-infected patients may be affected by a variety of renal disorders. Portugal has a high incidence of HIV2 infection and a low prevalence of HIV-infected patients under dialysis treatment. The aim of this study was to characterise the type of renal disease in Portuguese HIV-infected patients and to determine if HIV2 infection is associated to renal pathology. Only 60 of the 5158 HIV-infected patients followed in our hospital underwent renal biopsy. Clinical and laboratory data and the type of renal disease were reviewed. Male gender was predominant (76.7%), as was Caucasian race (78.3%). Mean age was 37.9±10.6 years. The majority had criteria for AIDS, 66% were on combined antiretroviral therapy and 18.3% were on dialysis. The predominant lesions were immunecomplex glomerulonephritis (n=19), tubulointerstitial nephropathy (n=12), focal segmental glomerulosclerosis(n=11), followed by HIVAN (n=8). Other patterns(amyloidosis, vasculitis, minimal change lesion) were observed. Only three patients were HIV2 infected, and presented diabetic nephropathy, acute tubular necrosis and tubulointerstitial nephritis. No correlations between clinical findings and renal pathology were found. In conclusion, renal disease in HIV patients has a broad spectrum, and renal biopsy remains the gold standard for establishing the diagnosis and guide treatment. Renal disease is not frequent in HIV2-infected patients, and, when present, is probably not directly associated with HIV infection.

Configuração de sistema de monitorização

O presente trabalho enquadra-se na área das redes de computadores, fazendo referência aos protocolos e ao conjunto de equipamentos e softwares necessários para a administração, controlo e monitorização desse tipo de infra-estruturas. Para a gestão de uma rede de dados, é essencial dispor de conhecimentos e documentação de nível técnico para representar da forma mais fiel possível a configuração da rede, seguindo passo a passo a interligação entre equipamentos existentes e oferecendo assim uma visão o mais fidedigna possível das instalações. O protocolo SNMP é utilizado em larga escala sendo praticamente um standard para a administração de redes baseadas na tecnologia TCP/IP. Este protocolo define a comunicação entre um administrador e um agente, estabelecendo o formato e o significado das mensagens trocadas entre ambos. Tem a capacidade de suportar produtos de diferentes fabricantes, permitindo ao administrador manter uma base de dados com informações relevantes da monitorização de vários equipamentos, que pode ser consultada e analisada por softwares NMS concebidos especialmente para a gestão de redes de computadores. O trabalho apresentado nesta dissertação teve como objectivo utilizar uma ferramenta NMS, para fazer a monitorização e a gestão da infra-estrutura de comunicações de forma que permitisse conhecer em tempo real o estado dos elementos de rede, ajudar no diagnóstico de possíveis problemas, instalados pela Nonius nos diversos navios da frota Douro Azul. O software NMS escolhido utiliza as potencialidades do protocolo SNMP para adquirir dados de monitorização de equipamentos de rede presentes na rede, bem como monitorizar redes remotas.

Effects of a Shortage of Imiglucerase on Three Patients with Type I Gaucher Disease

Background: Children with Gaucher disease type I (GD1) are usually treated with enzyme replacement therapy (ERT) at a dose of 30-60U/Kg/2W. Recently, due to an acute shortage supply of imiglucerase, a reduced dose or a reduced infusion frequency was recommended. Objective: To evaluate the effects of a reduced infusion frequency of imiglucerase over 15 months of follow-up. Patients and Methods: Three patients (1M:2F) were treated with ERT since a median age of 7 years (range 5-12). Only one had bone crisis and Erlenmeyer deformations. Median duration of treatment before dose reduction was 3 years (range 1-8). ERT resulted in total regression of symptoms, normalization of hematological parameters and progressive improvement of chitotriosidase in all patients. In August 2009 infusion schedule was changed from a media 45U/Kg every two weeks to every four weeks. Results: All patients remained asymptomatic and with no major change on hematological parameters except for the patient with bone crisis who presented subnormal platelet count. All patients showed an upward trend in chitotriosidase values. Comments: Although a longer follow-up is needed, is probable that even children completely stabilized can probably not be kept on lower doses even though the reduction of frequency of the infusions represent a lower social burden.

Simuladores Fisiológicos na avaliação metrológica de esfigmomanómetros

Este documento apresenta uma avaliação sobre o uso dos simuladores fisiológicos como padrão para avaliação metrológica dos esfigmomanómetros automáticos na estimação da pressão sanguínea pelo método não invasivo (PNI). O presente estudo procurou avaliar o enquadramento destes equipamentos com os procedimentos das normas e recomendações usadas para apreciação metrológica dos esfigmomanómetros digitais. No contexto da prática metrológica existente determinou-se a existência de uma oportunidade de melhoria nos processos relacionados. O trabalho procurou obter resposta a diversas questões, relacionando a medição da pressão pelo método não invasivo, com o uso dos simuladores fisiológicos, o contexto em que estes podem ser usados, as formas de simulação, as medições e os resultados, procurando a perspetiva metrológica como enquadramento. As recomendações existentes [1] [2] [3] [4] [5] [6] [7] [8], são muito claras nos procedimentos, validação e nos desvios permitidos para os monitores da tensão arterial (MTA), equipamento que permite a avaliação dos parâmetros fisiológicos do paciente, no entanto, quando se pretende avançar para outro domínio, como o do uso dos simuladores, em particular para a simulação da PNI, não existem recomendações ou normas tão claras, e não existe sobretudo um padrão de referência que imite a natureza dinâmica que caracteriza a pressão sanguínea. O trabalho procurou ainda estabelecer a ligação entre o método clássico de auscultação (o principio de determinação da PS), a técnica digital de medição e os simuladores, para uma melhor compreensão do que é a pressão sanguínea, e como relacionar a problemática da simulação e a de um padrão de referência. Neste trabalho estão ainda presentes abordagens a diversos tópicos, como as validações clínicas, acessórios, ou a metrologia e que influenciam no final os equipamentos e o contexto que se pretende avaliar. Os diversos equipamentos testados procuraram conter amostras diversificadas, quer para os MTA de uso profissional ou doméstico, assim como para os simuladores. A avaliação dos simuladores foi realizada contra um grupo de MTAs. Foi testada a influência na medição, causada pela mudança de acessórios, ou seja, nos resultados, merecendo consideração pela perspetiva metrológica. No resumo dos testes e do estudo sobre este tema, verificou-se que esta tipologia de equipamentos pode contribuir como complemento do processo de calibração típico (estático). Não constitui por si só um método alternativo, mas permitiu estimar possíveis limites de erro e desvio padrão a partir da observação dos resultados práticos, limites esses inferiores aos processos de validação clínica. Atendendo às particularidades, estimou-se como desvio aceitável um erro mais desvio padrão de 5 + 3 mmHg para o processo de simulação. Contudo considera-se ainda importante os testes adicionais em que o simulador permite, ao verificar as medidas de segurança implementadas no equipamento e a condição dos acessórios, que como verificado afetam os resultados. No entanto nem todos os simuladores se mostram adequados a este processo pelo que a qualidade da seleção do equipamento para este fim pode eventualmente reduzir ainda mais os possíveis limites.

Safety and skin delayed-type hypersensitivity response in vervet monkeys immunized with Leishmania donovani sonicate antigen delivered with adjuvants

In this study, we report on the safety and skin delayed-type hypersensitivity (DTH), responses of the Leishmania donovani whole cell sonicate antigen delivered in conjunction with alum-BCG (AlBCG), Montanide ISA 720 (MISA) or Monophosphoryl lipid A (MPLA) in groups of vervet monkeys. Following three intradermal injections of the inoculums on days 0, 28 and 42, safety and DTH responses were assessed. Preliminary tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) levels were also measured and these were compared with DTH. Only those animals immunized with alum-BCG reacted adversely to the inoculum by producing ulcerative erythematous skin indurations. Non-parametric analysis of variance followed by a post-test showed significantly higher DTH responses in the MISA+Ag group compared with other immunized groups (p < 0.001). The MPLA+Ag group indicated significantly lower DTH responses to the sonicate antigen compared with the AlBCG+Ag group. There was a significant correlation between the DTH and cytokine responses (p < 0.0001). Based on this study we conclude that Leishmania donovani sonicate antigen containing MISA 720 is safe and is associated with a strong DTH reaction following immunization.

Long-term biosignals visualization and processing

Thesis submitted in the fulfillment of the requirements for the Degree of Master in Biomedical Engineering

Cirurgia Obliterativa do Prolapso dos Órgãos Pélvicos - uma Opção ainda Válida?

Objective: To investigate the results of vaginal obliterate surgery in elderly women with pelvic organ prolapse. Design: observational retrospective study. Material and Methods: a total of 69 women with the diagnosis of pelvic prolapse were submitted to obliterative surgery in the urogynecology unit of a tertiary care hospital centre over the course of 8 years (2001 to 2008). The following data were collected from their clinical records: age, number of vaginal births, body mass index (BMI), hormone therapy, other existing diseases, type of prolapse and stage, anaesthetic risk score, duration of surgery, length of hospital stay, and short-term complications. Results: Of the 69 women studied, 31 were submitted to colpocleisis and the remaining 38 were managed by the LeFort technique. Mean age was 74.8 years with a standard deviation (sd) of 7.14 years. Average BMI was 26.2 (sd =3.76). Vaginal births were recorded in all patients. Only three patients were taking hormone therapy at the time of surgery. Sixty-three women were classified as having and anesthetic risk of II or III and 55 underwent local-regional anesthesia. Complications were reported in five cases, four of which in the first days after surgery. Nearly all were mild and resolved within the first 6 weeks. Conclusion: Complication rates appear to be low after obliterative surgery for pelvic organ prolapse in elderly women.

TESTING A SUBTYPE-SPECIFIC GP41 AMPLIFICATION METHOD FOR GENOTYPING INDIVIDUALS INFECTED BY HUMAN IMMUNODEFICIENCY VIRUS TYPE-1 IN THE BRAZILIAN POPULATION OF ITAJAÍ, SOUTH BRAZIL

The method used by YAGYU et al. for the subtype-specific polymerase chain reaction (PCR) amplification of the gp41 transmembrane region of the human immunodeficiency virus type-1 (HIV-1) env gene, was tested. HIV-1 proviral DNA from 100 infected individuals in Itajaí, South Brazil was used to analyze this method. Seventy individuals were determined according to this method as having PCR products at the expected size for subtypes B, C, D and F. Of these individuals, 26 (37.1%) were observed as having the expected amplification for subtype C, and 42 (60%) were observed as having the expected products for subtypes B and D. Of the subtype B and D amplicons, 16 (22.9%) were classified as subtype D, and 26 (37.1%) were classified as subtype B. Two individuals (2.9%) had amplicons that were observed after subtype F-specific amplification was performed. Sequencing and comparing the patient sequences to reference sequences confirmed the classification of sequences of subtypes C and B. However, sequences that were falsely determined as being D and F in the PCR assay were determined as being subtypes C and B, respectively, by sequence analysis. For those individuals from whom no amplified products were obtained, a low viral load that was indicated in their patient history may explain the difficulty in subtyping by PCR methods. This issue was demonstrated by the results of ANOVA when testing the effect of viral load on the success of PCR amplification. The alignment of the obtained sequences with HIV-1 reference sequences demonstrated that there is high intra-subtype diversity. This indicates that the subtype-specific primer binding sites were not conserved or representative of the subtypes that are observed in the Brazilian populations, and that they did not allow the correct classification of HIV-1 subtypes. Therefore, the proposed method by YAGYU et al. is not applicable for the classification of Brazilian HIV-1 subtypes.