Investigation of trypanothione synthetase of Leishmania infantum as a potential target for new anti-parasitic drugs

Dissertação para obtenção do Grau de Mestre em Biotecnologia

Incremental Recomputation of Pig Latin’s Nest and Unnest Operators

This master’s thesis addresses the maintenance of pre-computed structures, which store a frequent or expensive query, for the nested bag data type in the high level work-flow language Pig Latin. This thesis defines a model suitable to accommodate incremental expressions over nested bags on Pig Latin. Afterwards, the partitioned normal form for sets is extended with further restrictions, in order to accommodate the nested bag model, allow the Pig Latin nest and unnest operators revert each other, and create a suitable environment to the incremental computations. Subsequently, the extended operators – extended union and extended difference – are defined for the nested bag data model with the partitioned normal form for bags (PNF Bag) restriction, and semantics for the extended operators are given. Finally, incremental data propagation expressions are proposed for the nest and unnest operators on the data model proposed with the PNF Bag restriction, and the proof of correctness is given.

Evaluation of surrogate markers for human immunodeficiency virus infection among blood donors at the blood bank of "Hospital Universitário Regional Norte do Paraná", Londrina, PR, Brazil

This study evaluated the usefulness of the anti-HBc, hepatitis C virus antibodies (anti-HCV), human T cell lymphotropic virus I and II antibodies (anti-HTLV I/II), serologic tests for syphilis, and surface antigen of hepatitis B virus (HBsAg) as surrogate markers for the risk for HIV infection in 80,284 serum samples from blood donors from the Blood Bank of "Hospital Universitário Regional Norte do Paraná", Londrina, Paraná State, Brazil, analyzed from July 1994 to April 2001. Among 39 blood donors with positive serology for HIV, 12 (30.8%) were anti-HBc positive, 10 (25.6%) for anti-HCV, 1 (2.6%) for anti-HTLV I/I, 1 (2.6%) was positive for syphilis, and 1 (2.6%) for HBsAg. Among the donors with negative serology for HIV, these markers were detected in 8,407 (10.5%), 441 (0.5%), 189 (0.2%), 464 (0.6%), and 473 (0.6%) samples, respectively. The difference was statistically significant (p < 0.001) for anti-HBc and anti-HCV. Although the predictive positive value for these surrogate markers were low for HIV infection, the results confirmed the anti-HBc and anti-HCV as useful surrogate markers for HIV infection thus reinforcing the maintenance of them in the screening for blood donors contributing to the prevention of the small number of cases in which HIV is still transmitted by transfusion.

Evidence of active herpesvirus 6 (variant-A) infection in patients with lymphadenopathy in Belém, Pará, Brazil

A total of 323 patients with lymphadenopathy were selected in Belém, Brazil, between January 1996 and December 2001, and screened for the presence of human herpesvirus-6 (HHV-6) IgM- and- IgG antibodies by enzyme-linked immunosorbent assay (ELISA). When seroprevalence is analyzed by gender, similar rates are found for female (60.6%) and male (55.7%) individuals. Seventy-seven (23.8%) patients were HHV-6-IgM-and- IgG-positive (IgM+ subgroup), with positivity rates of 29.7% and 17.7% (p = 0.0007) for female- and male individuals, respectively. Sera from a subgroup (n = 120) of these subjects, with high HHV-6 antibody levels (either IgM+ or IgG+ reactivities), were subsequently processed for the presence of HHV-6 DNA by polymerase chain reaction (PCR)/nested PCR. Active infections (IgM+ and/or IgG+ high levels specific antibodies plus detection of viral DNA) were diagnosed in 20/77 (20.0%) and 8/43 (18.6%); subgroup of the 120 individuals suspected of having HHV-6 suggestive recent infection. All (n = 28) cases of active infection were found to be associated with HHV-6 variant-A (HHV-6A), as detectable by PCR/nested PCR, using variant-specific primer that amplify regions of 195 base pairs (bp) (HHV-6A) and 423 bp (HHV-6B). Rates of HHV-6 DNA detection between female and male patients were similar (p > 0.05) in the IgM+ and IgG+ groups: 20.4% versus 35.7% and 25.0% versus 13.0%, respectively. HHV-6 DNA was detected across < 5 through 41-50-year age-groups for patients whose serum samples were IgM+, with rates ranging from 7.7% (female subjects aged < 5 years) to 80.0% (male, 11-20 years). Among patients whose serological status was IgG+, HHV-6 DNA was detected in < 5, 6-10, 21-30 and > 50 age-groups at rates that ranged from 15.4% (male, < 5 years of age) to 100.0% (female aged 11-20 years). Swelling cervical lymph nodes were the most common sign, accounting for 9 (32.0%) cases in each gender group. Among patients (n = 28) with active infection by HHV-6A variant, duration of symptoms lasted 1-5 days in 35.7% of subjects, whereas in 64.3% of them the disease lasted 6-20 days. Our data suggest that it is worth seeking for HHV-6 infection whenever a patient (infant or adult) presents with lymphadenopathy as a prominent symptom in the course of an acute febrile illness.

Treatment of Vascular Lesions of the Tongue with Nd: YAG Laser

The treatment of vascular lesions of the tongue is a very challenging procedure since the maintenance of the lingual tissue is of critical importance. Numerous treatment options have been described in literature but the Nd:YAG Laser appears to be one of the safest therapeutic options. We described a successful treatment of vascular lesions of the tongue with an excellent clinical result after only one treatment session with the Nd:YAG laser, with conservation of the lingual tissue and its functionality.

Retrospective study of malaria prevalence and Anopheles genus in the area of influence of the binational Itaipu reservoir

The importance of hydroelectric dams beside the human interchange in the maintenance of malarious foci and the occurrence of the Anopheles genus on the Binational Itaipu Reservoir were the main points of this retrospective study. Data were collected from existing registrations at National, State and Municipal Health Departments and literature systematic overview, from January 1984 to December 2003. The occurrence of some outbreak of malaria, mainly by Plasmodium vivax, and the prevalence of species of the Anopheles genus different from Anopheles darlingi in the region are discussed. The malaria in the left bank of Paraná River is a focal problem, which must be approached locally through health, educational and social actions to prevent the continuity of outbreaks in the area. Concomitantly, it is necessary to plan and apply effective surveillance measures in the influence area of the Itaipu Reservoir.

Failure of nitric oxide production by macrophages and decrease in CD4+ T cells in oral paracoccidioidomycosis: possible mechanisms that permit local fungal multiplication

Paracoccidioidomycosis is a chronic granulomatous disease that induces a specific inflammatory and immune response. The participation of nitric oxide (NO), a product of the inducible nitric oxide synthase enzyme (iNOS), as an important fungicidal molecule against Paracoccidioides brasiliensis has been demonstrated. In order to further characterize the Oral Paracoccidioidomycosis (OP), we undertook an immunohistochemical study of iNOS+, CD45RO+, CD3+, CD8+, CD20+, CD68+ cells and mast cells. The samples were distributed in groups according to the number of viable fungi per mm². Our results showed weak immunolabeling for iNOS in the multinucleated giant cells (MNGC) and in most of the mononuclear (MN) cells, and the proportion of iNOS+ MN/MNGC cells in the OP were comparable to Control (clinically healthy oral tissues). Additionally, our analysis revealed a similarity in the number of CD4+ cells between the Control and the OP groups with higher numbers of fungi. These findings suggest that a low expression of iNOS and a decrease in the CD4+ T cells in OP may represent possible mechanisms that permit the local fungal multiplication and maintenance of active oral lesions.

Effects of Acute Autonomic Modulation on Atrial Conduction Delay and Local Electrograms Duration in Paroxysmal Atrial Fibrillation

Slowed atrial conduction may contribute to reentry circuits and vulnerability for atrial fibrillation (AF). The autonomic nervous system (ANS) has modulating effects on electrophysiological properties. However, complex interactions of the ANS with the arrhythmogenic substrate make it difficult to understand the mechanisms underlying induction and maintenance of AF. AIM: To determine the effect of acute ANS modulation in atrial activation times in patients (P) with paroxysmal AF (PAF). METHODS AND RESULTS: 16P (9 men; 59±14years) with PAF, who underwent electrophysiological study before AF ablation, and 15P (7 men; 58±11years) with atrioventricular nodal reentry tachycardia, without documentation or induction of AF (control group). Each group included 7P with arterial hypertension but without underlying structural heart disease. The study was performed while off drugs. Multipolar catheters were placed at the high right atrium (HRA), right atrial appendage (RAA), coronary sinus (CS) and His bundle area (His). At baseline and with HRA pacing (600ms, shortest propagated S2) we measured: i) intra-atrial conduction time (IACT, between RAA and atrial deflection in the distal His), ii) inter-atrial conduction time (interACT, between RAA and distal CS), iii) left atrial activation time (LAAT, between atrial deflection in the distal His and distal CS), iv) bipolar electrogram duration at four atrial sites (RAA, His, proximal and distal CS). In the PAF group, measurements were also determined during handgrip and carotid sinus massage (CSM), and after pharmacological blockade of the ANS (ANSB). AF was induced by HRA programmed stimulation in 56% (self-limited - 6; sustained - 3), 68.8% (self-limited - 6; sustained - 5), and 50% (self-limited - 5; sustained - 3) of the P, in basal, during ANS maneuvers, and after ANSB, respectively (p=NS). IACT, interACT and LAAT significantly lengthened during HRA pacing in both groups (600ms, S2). P with PAF have longer IACT (p<0.05), a higher increase in both IACT, interACT (p<0.01) and electrograms duration (p<0.05) with S2, and more fragmented activity, compared with the control group. Atrial conduction times and electrograms duration were not significantly changed during ANS stimulation. Nevertheless, ANS maneuvers increased heterogeneity of the local electrograms duration. Also, P with sustained AF showed longer interACT and LAAT during CSM. CONCLUSION: Atrial conduction times, electrograms duration and fractionated activity are increased in PAF, suggesting a role for conduction delays in the arrhythmogenic substrate. Acute vagal stimulation is associated with prolonged interACT and LAAT in P with inducible sustained AF and ANS modulation may influence the heterogeneity of atrial electrograms duration.

Desenvolvimento de sequenciador para um Problema de Roteamento de Veículos

Este projecto tem como objectivo a optimização das rotas dos técnicos de serviço após venda da Schmitt+Sohn Elevadores, associadas à realização das manutenções preventivas a cada elemento contratado à empresa (elevadores, escadas rolantes, etc). Como tal, é necessário fazer uma distribuição dos equipamentos que se encontram em carteira, por um dos técnicos que assegura a manutenção, pelos vários dias úteis de cada mês, e pelas horas de trabalho de cada dia. Apesar do técnico ter disponíveis, por dia, 8h de trabalho, apenas 6h podem ser preenchidas com manutenções preventivas. As 2h restantes são essencialmente para possíveis manutenções correctivas para as quais o técnico seja solicitado. Caso o técnico não seja contactado para resolver nenhuma avaria, essas horas podem ser utilizadas pelo mesmo para adiantar trabalho do dia seguinte, isto é, visitar já alguns dos próximos pontos de manutenção preventiva do dia seguinte, ou para compensar trabalho que esteja atrasado. De salientar que, para cada dia, as deslocações do técnico de qualquer local ao primeiro ponto de uma rota ou de regresso do último ponto de uma rota não são contabilizadas. O trabalho desenvolvido nesta dissertação pretende dar resposta ao problema apresentado pela Schmitt+Sohn Elevadores. Para isso foi desenvolvida uma heurística para a optimização das rotas dos técnicos. Esta é baseada no conceito de “vizinho mais próximo” que procura sempre o ponto que se apresenta mais perto do último ponto que foi adicionado à rota. Com base nesta metodologia, nos processos de escolha dos pontos que formam clusters, e na selecção dos pontos iniciais de cada uma das rotas diárias, a ferramenta de optimização resultante define as rotas diárias para que o percurso efectuado por cada técnico num mês seja o menor possível. São feitas alterações às rotas definidas inicialmente quando encontrados pontos de uma mesma entrada a serem visitados em dias diferentes. Isto obrigaria o técnico a fazer duas viagens ao mesmo local. Por fim, o resultado é apresentado num documento Word a ser utilizado pelo técnico como guia diário das suas deslocações aos equipamentos que necessitam de verificações periódicas. Os resultados obtidos foram comparados com as rotas que estavam a ser usadas pela empresa, tendo apresentado resultados de melhor qualidade, constatando-se a eficiência da solução criada pelo algoritmo proposto neste trabalho.

Inducibility of Atrial Fibrillation During Electrophysiologic Evaluation is Associated with Increased Dispersion of Atrial Refractoriness

The impact of atrial dispersion of refractoriness (Disp_A) in the inducibility and maintenance of atrial fibrillation (AF) has not been fully resolved. AIM: To study the Disp_A and the vulnerability (A_Vuln) for the induction of self-limited (<60 s) and sustained episodes of AF. METHODS AND RESULTS: Forty-seven patients with paroxysmal AF (PAF): 29 patients without structural heart disease and 18 with hypertensive heart disease. Atrial effective refractory period (ERP) was assessed at five sites--right atrial appendage and low lateral right atrium, high interatrial septum, proximal and distal coronary sinus. We compared three groups: group A - AF not inducible (n=13); group B - AF inducible, self-limited (n=18); group C - AF inducible, sustained (n=16). Age, lone AF, hypertension, left atrial and left ventricular (LV) dimensions, LV systolic function, duration of AF history, atrial flutter/tachycardia, previous antiarrhythmics, and Disp_A were analysed with logistic regression to determine association with A_Vuln for AF inducibility. The ERP at different sites showed no differences among the groups. Group A had a lower Disp_A compared to group B (47+/-20 ms vs 82+/-65 ms; p=0.002), and when compared to group C (47+/-20 ms vs 80+/-55 ms; p=0.008). There was no significant difference in Disp_A between groups B and C. By means of multivariate regression analysis, the only predictor of A_Vuln was Disp_A (p=0.04). Conclusion: In patients with PAF, increased Disp_A represents an electrophysiological marker of A_Vuln. Inducibility of both self-limited and sustained episodes of AF is associated with similar values of Disp_A. These findings suggest that the maintenance of AF is influenced by additional factors.

Biological implications of the phenotypic plasticity in the Schistosoma mansoni - Nectomys squamipes model

The water-rat Nectomys squamipes is mostly important non-human host in schistosomiasis mansoni transmission in Brazil, due to its susceptibility, high abundance and water-contact pattern. During experimental infection of N. squamipes with Schistosoma mansoni, adult worms show phenotypic plasticity. This finding led us to investigate whether biological behavior is also affected. This was assessed comparing the biological characteristics of four S. mansoni strains: BE (State of Belém do Pará), CE (State of Pernambuco), CMO (State of Rio Grande do Norte) and SJ (State of São Paulo) using laboratory-bred N. squamipes. The infection was monitored by determination of the pre-patent period, fecal egg output, egg viability, intestinal egg count and, infectivity rate. No biological modification was observed in these parameters. Overall results highlight that N. squamipes was susceptible to several S. mansoni strains, suggesting that it might contribute to the maintenance of schistosomiasis mansoni in Brazil.

Cytokine profile in Montenegro skin test of patients with localized cutaneous and mucocutaneous leishmaniasis

American tegumentary leishmaniasis presents as two major clinical forms: localized cutaneous leishmaniasis (LCL) and mucocutaneous leishmaniasis (MCL). The immune response in leishmaniasis is efficiently evaluated by the response to Leishmania antigen through the Montenegro skin test (MST). Both LCL and MCL present positive response to MST, indicating that the patients present cell-mediated immunity against the parasite - Leishmania. In spite of the presence of immunity in MCL, this is not sufficient to stop disease progression and prevent resistance to treatment. In this study we demonstrated interleukin (IL) 2, 4, 5 and interferon (IFN) gamma expression in biopsies of MST of ten patients with American tegumentary leishmaniasis. The obtained results were compared between LCL (n = 5) and MCL (n = 5) patients. The MST of MCL patients displayed a higher expression of IL-2, IL-4 and IL-5, in comparison to LCL. There was no significant difference in IFN-gamma expression between groups. The obtained results suggest the role of IL-4 and IL-5 in the maintenance of the immunopathogenic mechanism of the destructive lesions that characterize MCL.

Process engineering of liver cells for drug testing applications

Dissertação para obtenção do Grau de Doutor em Bioengenharia

Inspeção e reabilitação de estruturas de betão armado

O setor da reabilitação em Portugal tem vindo a crescer consideravelmente ao longo dos últimos anos, demonstrando uma consciencialização da sociedade e das entidades competentes para definição de estratégias e planeamento, para uma manutenção das estruturas atempada de modo a prevenir patologias que provocam a sua degradação. O presente trabalho pretende fazer uma síntese das técnicas de proteção e reparação das estruturas de betão armado, efetuando também ensaios para determinação das causas das anomalias que levaram à sua reparação. O projeto de reabilitação deve seguir as recomendações indicadas nos Eurocódigos e norma EN1504 sobre produtos e sistemas para a proteção e reparação das estruturas de betão. Segundo esta norma explicam – se as principais causas de deterioração das estruturas de betão armado e os sintomas que estão na base dessa deterioração. Desta forma são descritos os métodos de intervenção, explicando o seu funcionamento e modo mais adequado de utilização com indicação dos produtos adequados para cada intervenção. Para aplicação do referido nos parágrafos anteriores desenvolveu-se uma proposta de fichas de inspeção, através de levantamento fotográfico e registo das anomalias e suas causas e ensaios não destrutivos de modo a perceber o estado atual da estrutura para uma melhor compreensão da metodologia de reparação a seguir

Análise da viabilidade económica da implementação de uma central de micro-cogeração no setor da agropecuária

O crescente aumento do consumo energético das sociedades desenvolvidas e emergentes, motivado pelo progresso económico e social, tem induzido a procura de alternativas focalizadas nas energias renováveis, que possam contribuir para assegurar o fornecimento de energia sem agravar o consumo de combustíveis fósseis e a emissão de gases com efeito de estufa. Nesse sentido, a produção de energia eléctrica a partir do gás metano resultante da estabilização anaeróbia de efluentes tem vindo a ser estudada e praticada desde finais do século XIX, tendo assumido maior expressão a partir dos anos 70 do século XX, na sequência das primeiras crises petrolíferas. As instalações agropecuárias reúnem dois fatores chave para o sucesso do aproveitamento energético do biogás produzido no tratamento dos efluentes: por um lado, produzem matéria-prima com potencial energético – dejeto animal com um potencial enorme de criação de biogás quando procedido de tratamento anaeróbio - e, por outro, necessitam de energia eléctrica para o funcionamento dos equipamentos electromecânicos e de calor para a manutenção das instalações. A valorização energética do biogás produzido na estabilização anaeróbia dos efluentes agro-pecuários, para além de permitir obter um retorno financeiro, que contribui para o equilíbrio dos custos de investimento e de exploração, contribui igualmente para a redução das emissões de gases com efeito de estufa, como o dióxido de carbono e o metano, e para a segurança de abastecimento energético à instalação, na medida em que assegura a alimentação de energia eléctrica em caso de falha no fornecimento pela rede nacional. A presente dissertação apresenta um contributo para estudos a desenvolver por proprietários de agropecuárias, cooperativas regionais do setor da agropecuária, empresas de projecto e estudantes de Engenharia, constituído por uma compilação da informação mais relevante associada à estabilização anaeróbia de efluentes e à valorização energética do biogás produzido. Com base em informação referente ao número real de animais existentes em Portugal, este trabalho pretende fazer ver a essas entidades que o aproveitamento energético do biogás é viável e útil para o país. Com a criação de uma aplicação informática de análise económica de investimento, provar que o investimento em pequenas propriedades, com apenas 80 cabeças normais, pode obter um retorno financeiro razoável, com um prazo de recuperação do investimento bastante baixo, aproveitando um recurso que caso contrário será desperdiçado e poluirá o ambiente.