Etiological agents of diarrhea in patients infected by the human immunodeficiency virus-1: a review

Despite the importance of understanding the epidemiology of agents responsible for infectious diarrhea in human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) population, the number of articles about this subject is relatively few. The current article summarizes published data on bacterial, fungal, viral and parasitic enteropathogens in the HIV/AIDS seropositive subjects in different countries, regions and localities. In general, there is a great difference in the frequencies of etiological agents due to factors which include immune status, geographical location, climate and socioeconomic conditions. It is important to stress that a great prevalence of infection by emergent agents has been reported in the more advanced stages of AIDS. Therefore, to establish specific treatment depends directly on knowledge of these agents and risk factors associated to their distribution. Moreover, the colonization by potential pathogenic agents verified in these individuals is high thus implicating that they act as carriers. Finally, public health measures of control and prevention must take into consideration the regional previously identified enteropathogens, especially in areas where HIV prevalence is high.

Perinatal Bacterial Infection: Screenning of Vertical Transmitted Infections

Perinatal bacterial infection may be caused by any microorganism colonizing the vaginal tract. Neonatologists and paediatricians are especially concerned about group B Stretpococcus (GBS). However, Enterobactereacea, mainly E.coli and Proteus, are also responsible for infection. GBS screening may be accomplished in over 90% of pregnant women. In our maternity in 2007-2008, 85% of the mothers had been screened. Screening and prophylaxis were responsible for a decreasing incidence of neonatal infection - from 0.6/1000 to 0.15/1000 live births in Portugal, from 2002 to 2007. However there are some difficulties related to screening. In the second Portuguese study 16/57 NB with early-onset infection (28%) were born to “negative” mothers. Several factors illustrate how difficult is to draw national screening policies: a wide range of carrier’s state rate throughout a country - in Portugal from 12% to 30%. The success of any screening policy may also be affected by additional technical and organizational problems. In countries where home delivery is a tradition or a trend intrapartum GBS prophylaxis requires a very well organized assistance.. Moreover factors usually accepted as protective are not so effective. In the Portuguese study 24/57 infected newborns (42%) were delivery by caesarean section. Another subject deals with the workload in the postnatal ward generated by deficient compliance to the guidelines a problem not confirm by a study of our group. Decreasing the importance of GBS, highlight the importance of E. coli in perinatal infection. From the 16 340 registrations of the National Registry 1676 were newborns with mother-related infection. Applying the same reasoning to E.coli as to GBS and Listeria monocytogenes – that is considering all of them are of maternal origin - 6.7% of these infections were due to E. coli, 4.6% to SGB and 0.5% to Listeria monocytogenes. In conclusion screening and prophylaxis may be not the best way to prevent all GBS neonatal infections but by now it is the only available procedure. The other bacteria continue to demand a high suspicion level and immediate intervention.

Utilização do software Scratch no ensino das ciências da natureza e da matemática com alunos portadores de dislexia

O estudo que apresentamos tem como objetivos de investigação contribuir para o desenvolvimento global das crianças com Necessidades Educativas Especiais, nomeadamente, portadoras de Dislexia, designadamente em aulas onde são lecionados conteúdos de Ciências da Natureza e de Matemática, e conceber um recurso didático apoiado nas TIC que seja adequado à utilização em sala de aula por alunos com dislexia. Neste sentido, o estudo foi orientado pelas seguintes questões de investigação: A utilização do Scratch é adequada e exequível no trabalho quotidiano de alunos portadores de Dislexia? e Em que medida o Scratch pode contribuir para um melhor envolvimento dos alunos portadores de Dislexia na realização das tarefas propostas? O presente trabalho segue uma metodologia de carácter qualitativo, centrando-se no estudo de caso. Tentou-se dar uma resposta à reduzida utilização do software Scratch no ensino das Ciências da Natureza e Matemática como instrumentos viáveis para uma aprendizagem de sucesso em alunos portadores de Dislexia. Este estudo foi aplicado a três alunos do 4º ano, com este diagnóstico, de um agrupamento de escolas do concelho de Viseu. Para dar resposta às questões de investigação supracitadas foram implementadas e projetadas duas atividades no software Scratch para serem desenvolvidas em duas situações formativas contextualizadas. O subdomínio que se pretendeu trabalhar foi “A importância da água para os seres vivos” articulando com o conteúdo dos volumes. Os dados foram recolhidos através da análise dos Programas Educativos Individuais de cada um dos alunos participantes, das entrevistas às docentes de Educação Especial, de gravações áudio, fotografias, trabalhos dos alunos e os registos da professora investigadora. Os resultados obtidos permitiram caracterizar o modo como os alunos, portadores de dislexia, se envolveram com o Scratch na aquisição de conhecimento. Permitiram ainda demonstrar aos professores que é possível construir atividades nesta ferramenta envolvendo conteúdos curriculares interdisciplinares.

Evaluation of GBV-C / HVG viremia in HIV-infected women

The present study aimed at standardizing a real-time quantitative polymerase chain reaction assay to evaluate the presence of GBV-C/HGV RNA. A "TaqMan" assay using primers and probe derived from the 5¢ NCR region was developed and validated. Two hundred and fifty-three plasma samples from HIV-infected women were tested for GBV-C viremia and antibody against the envelope protein 2. GBV-C RNA was detected in 22.5% of the patients whereas the antibody was identified in 25.3% of the cohort. Detection of viral RNA and of antibodies was mutually exclusive. Viral loads showed a mean of 1,777 arbitrary units / mL, being 1.1 and 13,625 arbitrary units / mL respectively the lowest and highest values measured. We conclude that the real-time quantitative polymerase chain reaction method developed is appropriate for the investigation of GBV-C RNA since it was shown to be highly specific and sensitive, as well as requiring few steps, preventing contamination and providing additional information as to the relative viremia of carriers, a parameter that must be included in studies evaluating the co-factors influencing the clinical outcome of HIV/AIDS.

Dimensionar a logística interna da fábrica Toyota Ovar para a produção de nova viatura

O projeto foi realizado na Toyota (divisão fabril de Ovar), no ano letivo 2014/2015, teve por base um estágio extracurricular que foi realizado para que desta forma tomasse contacto com uma realidade concreta e absoluta no contexto empresarial. O estágio serviu, principalmente, como uma experiência na compreensão do funcionamento de uma indústria de automóveis. Este projeto desenvolve-se no âmbito do 2ºano do Mestrado de Logística e apresenta os contributos desenvolvidos no departamento de logística para a reorganização da mesma para a produção de um novo modelo automóvel. Atualmente, a Toyota, para os modelos de automóveis que produz, recebe os componentes em CKD (Completed Knocked Down) provenientes do Japão em lotes de cinco unidades. Estes lotes são armazenados no armazém, de onde só saem quando for dada ordem para serem produzidos. Cada lote tem várias caixas, que são colocadas à vez numa zona comum de abertura. São retidos os componentes das caixas CKD e colocados nos transportadores próprios dos postos correspondentes (supermercado de linha). Isto significa que sempre que é solicitado uma viatura de um determinado lote, são obrigados a produzir as cinco unidades. Isto significa que as restantes unidades do lote terão que ficar em parque aberto a aguardar venda futura, para além de ter um stock desnecessário, estas unidades ficam vulneráveis e sofrem desgaste, pois estão expostas à intempérie, causando graves problemas de qualidade e respetivos custos associados. Recentemente, foi decidida a produção do modelo Land Cruiser 70 na unidade industrial de Ovar, Portugal, o que obrigou a uma reorganização da produção e processos logísticos para a sua produção. Os objetivos que a empresa pretende com a entrada de um novo projeto Land Cruiser 70, são os seguintes: Alterar o sistema de abastecimento, implementando o abastecimento unitário em toda a fábrica para o novo modelo, garantindo o “one by one production”; Definir o funcionamento do processo logístico desde a chegada dos componentes até ficarem disponíveis no supermercado de linha. Para isso será necessário dimensionar armazém e zona de abertura, definir layouts e quantidade de stocks, bem como definir sistemas de comunicação entre abertura e supermercado; Verificar as peças que vão fazer parte do chassi por posto e contabilizar os dolly´s1 necessários; Fazer o layout de abastecimento da linha do chassi; Calcular o tempo de abertura, de abastecimento e de picking do chassi e de triming. Este projeto tem em conta a política de melhoria contínua de forma a conseguir satisfazer o cliente (ser eficaz) utilizando cada vez menos recursos quer físicos quer humanos (ser eficiente, detetar e eliminar os desperdícios). O presente projeto foi inserido na equipa de melhoria contínua da logística, maioritariamente no chassi. Em termos globais, os objetivos propostos neste estágio foram atingidos com êxito.

PREVALENCE OF INTESTINAL PARASITISM AND ASSOCIATED SYMPTOMATOLOGY AMONG HEMODIALYSIS PATIENTS

Intestinal parasites are an important cause of morbidity and mortality. Immunocompromised individuals may develop more severe forms of these infections. Taking into account the immunity impairment in patients suffering from chronic renal failure (CRF), we will determine the prevalence and associated symptoms of intestinal parasites in these patients. Controls without CRF were used for comparison. Stool samples were collected and processed for microscopic identification of parasites using the Formalin-ether concentration method. For Cryptosporidium diagnosis, the ELISA technique was used. One hundred and ten fecal samples from hemodialysis patients were analyzed, as well as 86 from a community group used as control group. A result of 51.6% of intestinal parasites was observed in hemodialysis patients and 61.6% in the control group. Cryptosporidium and Blastocystis were the most common infections in patients with CRF (26.4% and 24.5%, respectively). Blastocystis was the most common infection in the control group (41.9%), however no individual was found positive for Cryptosporidium. Among the CRF patients, 73.6% were symptomatic, 54.3% of these tested positive for at least one parasite, in contrast to 44.8% in asymptomatic patients (p = 0.38). The most common symptoms in this group were flatulence (36.4%), asthenia (30.0%) and weight loss (30.0%). In the control group, 91.9% were symptomatic, 60.8% of these tested positive for at least one parasite, in contrast to 71.4% in asymptomatic patients (p = 0.703). A significant difference between the two groups was observed with regard to symptoms, with bloating, postprandial fullness, and abdominal pain being more frequent in the control group than in the hemodialysis group (all p < 0.05). Comparing symptomatic with asymptomatic, there was no association in either group between symptoms or the prevalence of parasitic infection, nor with the type of parasite or with multiple parasitic infections. Patients with chronic renal failure are frequent targets for renal transplantation, which as well as the inherent immunological impairment of the disease itself, results in immunosuppression by medication. For this reason, carriers of intestinal parasites with pathogenic potential can develop serious clinical complications influencing the success of transplantation. This fact, coupled with the high prevalence of intestinal parasites and the dissociation between symptoms and infection in CRF patients, suggests that the stool test should be incorporated in routine propedeutics. Furthermore, preventive measures for the acquisition of parasites through the fecal-oral contamination route should be introduced.

Three transducers for one photodetector: essays for optical communications

Dissertation presented to obtain the PhD degree in Electrical and Computer Engineering - Electronics

Anopheles (Kerteszia) cruzii (DIPTERA: CULICIDAE) IN PERIDOMICILIARY AREA DURING ASYMPTOMATIC MALARIA TRANSMISSION IN THE ATLANTIC FOREST: MOLECULAR IDENTIFICATION OF BLOOD-MEAL SOURCES INDICATES HUMANS AS PRIMARY INTERMEDIATE HOSTS

Anopheles (Kerteszia) cruzii has been implicated as the primary vector of human and simian malarias out of the Brazilian Amazon and specifically in the Atlantic Forest regions. The presence of asymptomatic human cases, parasite-positive wild monkeys and the similarity between the parasites infecting them support the discussion whether these infections can be considered as a zoonosis. Although many aspects of the biology of An. cruzii have already been addressed, studies conducted during outbreaks of malaria transmission, aiming at the analysis of blood feeding and infectivity, are missing in the Atlantic Forest. This study was conducted in the location of Palestina, Juquitiba, where annually the majority of autochthonous human cases are notified in the Atlantic Forest of the state of São Paulo. Peridomiciliary sites were selected for collection of mosquitoes in a perimeter of up to 100 m around the residences of human malaria cases. The mosquitoes were analyzed with the purpose of molecular identification of blood-meal sources and to examine the prevalence of Plasmodium. A total of 13,441 females of An. (Ker.) cruzii were collected. The minimum infection rate was calculated at 0.03% and 0.01%, respectively, for P. vivax and P. malariae and only human blood was detected in the blood-fed mosquitoes analyzed. This data reinforce the hypothesis that asymptomatic human carriers are the main source of anopheline infection in the peridomiciliary area, making the probability of zoonotic transmission less likely to happen.

The Role of the Humoral Immune Response in the Molecular Evolution of the Envelope C2, V3 and C3 Regions in Chronically HIV-2 Infected Patients

BACKGROUND: This study was designed to investigate, for the first time, the short-term molecular evolution of the HIV-2 C2, V3 and C3 envelope regions and its association with the immune response. Clonal sequences of the env C2V3C3 region were obtained from a cohort of eighteen HIV-2 chronically infected patients followed prospectively during 2-4 years. Genetic diversity, divergence, positive selection and glycosylation in the C2V3C3 region were analysed as a function of the number of CD4+ T cells and the anti-C2V3C3 IgG and IgA antibody reactivity RESULTS: The mean intra-host nucleotide diversity was 2.1% (SD, 1.1%), increasing along the course of infection in most patients. Diversity at the amino acid level was significantly lower for the V3 region and higher for the C2 region. The average divergence rate was 0.014 substitutions/site/year, which is similar to that reported in chronic HIV-1 infection. The number and position of positively selected sites was highly variable, except for codons 267 and 270 in C2 that were under strong and persistent positive selection in most patients. N-glycosylation sites located in C2 and V3 were conserved in all patients along the course of infection. Intra-host variation of C2V3C3-specific IgG response over time was inversely associated with the variation in nucleotide and amino acid diversity of the C2V3C3 region. Variation of the C2V3C3-specific IgA response was inversely associated with variation in the number of N-glycosylation sites. CONCLUSION: The evolutionary dynamics of HIV-2 envelope during chronic aviremic infection is similar to HIV-1 implying that the virus should be actively replicating in cellular compartments. Convergent evolution of N-glycosylation in C2 and V3, and the limited diversification of V3, indicates that there are important functional constraints to the potential diversity of the HIV-2 envelope. C2V3C3-specific IgG antibodies are effective at reducing viral population size limiting the number of virus escape mutants. The C3 region seems to be a target for IgA antibodies and increasing N-linked glycosylation may prevent HIV-2 envelope recognition by these antibodies. Our results provide new insights into the biology of HIV-2 and its relation with the human host and may have important implications for vaccine design.

Enterobacteriaceae ISOLATES FROM THE ORAL CAVITY OF WORKERS IN A BRAZILIAN ONCOLOGY HOSPITAL

The evaluation of workers as potential reservoirs and disseminators of pathogenic bacteria has been described as a strategy for the prevention and control of healthcare-associated infections (HAIs). The aim of this study was to evaluate the presence of Enterobacteriaceae in the oral cavity of workers at an oncology hospital in the Midwest region of Brazil, as well as to characterize the phenotypic profile of the isolates. Saliva samples of 294 workers from the hospital’s healthcare and support teams were collected. Microbiological procedures were performed according to standard techniques. Among the participants, 55 (18.7%) were colonized by Enterobacteriaceae in the oral cavity. A total of 64 bacteria were isolated, including potentially pathogenic species. The most prevalent species was Enterobacter gergoviae (17.2%). The highest rates of resistance were observed for β-lactams, and 48.4% of the isolates were considered multiresistant. Regarding the enterobacteria isolated, the production of ESBL and KPC was negative. Nevertheless, among the 43 isolates of the CESP group, 51.2% were considered AmpC β-lactamase producers by induction, and 48.8% were hyper-producing mutants. The significant prevalence of carriers of Enterobacteriaceae and the phenotypic profile of the isolates represents a concern, especially due to the multiresistance and production of AmpC β-lactamases.

PREVALENCE OF INTESTINAL PARASITES AMONG FOOD HANDLERS OF SARI, NORTHERN IRAN

Parasitic infection is highly prevalent throughout the developing countries of the world. Food handlers are a potential source of infection for many intestinal parasites and other enteropathogenic infections as well. The aim of this study was to determine the prevalence of intestinal parasite carriers among food handlers attending the public health center laboratory in Sari, Northern Iran for annual check-up. The study was performed from August 2011 through February 2012. Stool samples were collected from 1041 male and female food handlers of different jobs aged between 18 to 63 years and were examined following standard procedures. Sociodemographic, environmental and behavioral data analysis of the food handlers were recorded in a separate questionnaire. Intestinal parasites were found in 161 (15.5%) of the studied samples. Seven species of protozoan or helminth infections were detected. Most of the participants were infected with Giardia lamblia (53.9%) followed by Blastocystis hominis (18%), Entamoeba coli (15.5%), Entamoeba histolytica/dispar (5.5%), Cryptosporidium sp. (3.1%), Iodamoeba butschlii (3.1%) and Hymenolepis nana (1.9%) as the only helminth infection. The findings emphasized that food handlers with different pathogenic organisms may predispose consumers to significant health risks. Routine screening and treatment of food handlers is a proper tool in preventing food-borne infections.

Development of new oxygen therapeutics using fluorinated ionic liquids

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau Mestre em Engenharia Biomédica

Evolution of the Human Immunodeficiency Virus Type 2 Envelope in the First Years of Infection is Associated with the Dynamics of the Neutralizing Antibody Response

Background: Differently from HIV-1, HIV-2 disease progression usually takes decades without antiretroviral therapy and the majority of HIV-2 infected individuals survive as elite controllers with normal CD4+ T cell counts and low or undetectable plasma viral load. Neutralizing antibodies (Nabs) are thought to play a central role in HIV-2 evolution and pathogenesis. However, the dynamic of the Nab response and resulting HIV-2 escape during acute infection and their impact in HIV-2 evolution and disease progression remain largely unknown. Our objective was to characterize the Nab response and the molecular and phenotypic evolution of HIV-2 in association with Nab escape in the first years of infection in two children infected at birth. Results: CD4+ T cells decreased from about 50% to below 30% in both children in the first five years of infection and the infecting R5 viruses were replaced by X4 viruses within the same period. With antiretroviral therapy, viral load in child 1 decreased to undetectable levels and CD4+ T cells recovered to normal levels, which have been sustained at least until the age of 12. In contrast, viral load increased in child 2 and she progressed to AIDS and death at age 9. Beginning in the first year of life, child 1 raised high titers of antibodies that neutralized primary R5 isolates more effectively than X4 isolates, both autologous and heterologous. Child 2 raised a weak X4-specific Nab response that decreased sharply as disease progressed. Rate of evolution, nucleotide and amino acid diversity, and positive selection, were significantly higher in the envelope of child 1 compared to child 2. Rates of R5-to-X4 tropism switch, of V1 and V3 sequence diversification, and of convergence of V3 to a β-hairpin structure were related with rate of escape from the neutralizing antibodies. Conclusion: Our data suggests that the molecular and phenotypic evolution of the human immunodeficiency virus type 2 envelope are related with the dynamics of the neutralizing antibody response providing further support for a model in which Nabs play an important role in HIV-2 pathogenesis.

ISOLATION AND MOLECULAR IDENTIFICATION OF POTENTIALLY PATHOGENIC Escherichia coli AND Campylobacter jejuni IN FERAL PIGEONS FROM AN URBAN AREA IN THE CITY OF LIMA, PERU

SUMMARY Feral pigeons (Columbia livia) live in close contact with humans and other animals. They can transmit potentially pathogenic and zoonotic agents. The objective of this study was to isolate and detect strains of diarrheagenic Escherichia coli and Campylobacter jejuniof urban feral pigeons from an area of Lima, Peru. Fresh dropping samples from urban parks were collected for microbiological isolation of E. coli strains in selective agar, and Campylobacterby filtration method. Molecular identification of diarrheagenic pathotypes of E.coliand Campylobacter jejuni was performed by PCR. Twenty-two parks were sampled and 16 colonies of Campylobacter spp. were isolated. The 100% of isolates were identified as Campylobacter jejuni. Furthermore, 102 colonies of E. coli were isolated and the 5.88% resulted as Enteropathogenic (EPEC) type and 0.98% as Shiga toxin-producing E. coli (STEC). The urban feral pigeons of Lima in Peru can act as a reservoir or carriers of zoonotic potentially pathogenic enteric agents.

Modelação fenomenológica de um motor Diesel marítimo para avaliar efeitos da mudança de combustível

Dissertação para obtenção do Grau de Mestre em Engenharia Mecânica