C4d Detection in Renal Allograft Biopsies: Immunohistochemistry vs. Immunofluorescence

Introduction. Peritubular capillary complement 4d staining is one of the criteria for the diagnosis of antibody-mediated rejection, and research into this is essential to kidney allograft evaluation. The immunofluorescence technique applied to frozen sections is the present gold-standard method for complement 4d staining and is used routinely in our laboratory. The immunohistochemistry technique applied to paraffin-embedded tissue may be used when no frozen tissue is available. Material and Methods. The aim of this study is to evaluate the sensitivity and specificity of immunohistochemistry compared with immunofluorescence. We describe the advantages and disadvantages of the immunohistochemistry vs. the immunofluorescence technique. For this purpose complement 4d staining was performed retrospectively by the two methods in indication biopsies (n=143) and graded using the Banff 07 classification. Results. There was total classification agreement between methods in 87.4% (125/143) of cases. However, immunohistochemistry staining caused more difficulties in interpretation, due to nonspecific staining in tubular cells and surrounding interstitium. All cases negative by immunofluorescence were also negative by immunohistochemistry. The biopsies were classified as positive in 44.7% (64/143) of cases performed by immunofluorescence vs. 36.4% (52/143) performed by immunohistochemistry. Fewer biopsies were classified as positive diffuse in the immunohistochemistry group(25.1% vs. 31.4%) and more as positive focal (13.2% vs. 11.1%). More cases were classified as negative by immunohistochemistry (63.6% vs. 55.2%). Study by ROC curve showed immunohistochemistry has a specificity of 100% and a sensitivity of 81.2% in relation to immunofluorescence (AUC: 0.906; 95% confidence interval: 0.846-0.949; p=0.0001). Conclusions. The immunohistochemistry method presents an excellent specificity but lower sensitivity to C4d detection in allograft dysfunction. The evaluation is more difficult, requiring a more experienced observer than the immunofluorescence method. Based on these results, we conclude that the immunohistochemistry technique can safely be used when immunofluorescence is not available.

Therapeutic Effects of Allium sativum and Allium cepa in Schistosoma mansoni experimental infection

The effects of both garlic (Allium sativum) and onion (Allium cepa) on some biochemical parameters in Schistosoma mansoni infected mice individually and mixed either with or without the currently used drug, praziquantel (PZQ) were investigated. These involved some immunological parameters, namely IgM, IgG, interleukins 2 and 6 (IL-2 and 6) and tumor necrosis factor (TNF-α), some antioxidant enzymes [catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPX)]. In addition, parasitological and histopathological investigations were performed. No changes were observed in the normal control mice treated with dry extract of onion or garlic, individually or mixed, with or without PZQ, compared to the normal healthy control group. Infection with S. mansoni showed an increase in IgG, IgM, IL-2, IL-6, TNF-α and catalase enzyme, accompanied with a decrease in GPX and SOD antioxidant enzyme activities. Remarkable amelioration was noticed in the levels of all the measured parameters in S. mansoni infected mice after administration of the studied extracts. Moreover a significant reduction in worm burden, hepatic and intestinal eggs and oogram count was noticed which was reflected in normalization of liver architecture.

Induction of phagocytic activity and nitric-oxide production in natural populations of Trypanosoma Cruzi I and II from the state of Paraná, Brazil

Twelve strains of Trypanosoma cruzi isolated from wild reservoirs, triatomines, and chronic chagasic patients in the state of Paraná, southern Brazil, and classified as T. cruzi I and II, were used to test the correlation between genetic and biological diversity. The Phagocytic Index (PI) and nitric-oxide (NO) production in vitro were used as biological parameters. The PI of the T. cruzi I and II strains did not differ significantly, nor did the PI of the T. cruzi strains isolated from humans, triatomines, or wild reservoirs. There was a statistical difference in the inhibition of NO production between T. cruzi I and II and between parasites isolated from humans and the strains isolated from triatomines and wild reservoirs, but there was no correlation between genetics and biology when the strains were analyzed independently of the lineages or hosts from which the strains were isolated. There were significant correlations for Randomly Amplified Polymorphic Deoxyribonucleic acid (RAPD) and biological parameters for T. cruzi I and II, and for humans or wild reservoirs when the lineages or hosts were considered individually.

Thermal shock – and its perfect imperfections

Dissertação de Mestrado em Arte e Ciência do Vidro

The development and implementation of math projects in a HEI: expectations, objectives, experiences and analysis

An overwhelming problem in Math Curriculums in Higher Education Institutions (HEI), we are daily facing in the last decade, is the substantial differences in Math background of our students. When you try to transmit, engage and teach subjects/contents that your “audience” is unable to respond to and/or even understand what we are trying to convey, it is somehow frustrating. In this sense, the Math projects and other didactic strategies, developed through Learning Management System Moodle, which include an array of activities that combine higher order thinking skills with math subjects and technology, for students of HE, appear as remedial but important, proactive and innovative measures in order to face and try to overcome these considerable problems. In this paper we will present some of these strategies, developed in some organic units of the Polytechnic Institute of Porto (IPP). But, how “fruitful” are the endless number of hours teachers spent in developing and implementing these platforms? Do students react to them as we would expect? Do they embrace this opportunity to overcome their difficulties? How do they use/interact individually with LMS platforms? Can this environment that provides the teacher with many interesting tools to improve the teaching – learning process, encourages students to reinforce their abilities and knowledge? In what way do they use each available material – videos, interactive tasks, texts, among others? What is the best way to assess student’s performance in these online learning environments? Learning Analytics tools provides us a huge amount of data, but how can we extract “good” and helpful information from them? These and many other questions still remain unanswered but we look forward to get some help in, at least, “get some drafts” for them because we feel that this “learning analysis”, that tackles the path from the objectives to the actual results, is perhaps the only way we have to move forward in the “best” learning and teaching direction.

A eficiência energética nas redes de telecomunicações e tecnologias de informação e comunicação

A presente tese pretende fazer uma abordagem ao crescimento dos consumos de energia elétrica, que se tem verificado, nos últimos anos, no setor das telecomunicações e das tecnologias de informação; devido ao constante crescimento das redes, dos equipamentos a ela ligados e do tráfego que nelas transita. Num contexto de globalização da economia, no qual, as redes de telecomunicações e de energia elétrica são dois dos maiores contribuintes, a presente tese procura encontrar enquadramentos e soluções para um dos maiores desafios que a humanidade enfrenta atualmente, e que em parte, é consequente dessa globalização: encontrar novas fontes e formas de utilização da energia, -particularmente da energia elétrica - para que a humanidade continue a usufruir, de uma forma sustentável, dos benefícios que a mesma proporciona. Na primeira parte, procura-se fazer uma abordagem que utiliza fontes de informação e conhecimento, do mercado global, nomeadamente, entidades reguladoras e normalizadoras, operadores, fornecedores de tecnologias e consumidores, que abrangessem os três maiores mercados mundiais – União Europeia, Estados Unidos da América e Ásia-Pacífico. Considerou-se fundamental fazê-lo, por se tratar de uma dissertação no âmbito de um Mestrado com o selo de garantia EUR-ACE. Ao longo da dissertação analisou-se a temática da eficiência energética nas redes de telecomunicações e das tecnologias de informação e comunicação, um tema cada vez mais pertinente, já que o número de pessoas com ligações à Internet, já supera os 3 mil milhões, e as redes passaram a ser o meio por onde são transmitidos, a cada segundo, terabytes de sinais de voz, dados e vídeo. Procurou-se encontrar as linhas de orientação que estão a ser traçadas, para otimizar os consumos energéticos, de um complexo sistema convergente de redes e serviços, formado por entidades reguladoras e normalizadoras, operadores, fornecedores de tecnologias e consumidores, onde nem sempre as fronteiras estão perfeitamente definidas. Perante a constatação da realidade exposta, analisou-se as políticas energéticas desenvolvidas nos últimos anos, pelos vários players do mercado das telecomunicações, das tecnologias de informação e dos sistemas elétricos de energia bem como algumas métricas e objetivos comumente aceites. viii São analisados os contributos das partes interessadas, para o desenvolvimento de políticas energéticas eficazes, por forma a permitirem uma implementação, que considere o funcionamento dos equipamentos como um todo, e não de uma forma isolada como tradicionalmente o assunto era abordado. As especificidades na forma como funcionam as redes de telecomunicações e respetivos equipamentos, são expostas sobre várias óticas, comprovando-se que a temática da eficiência energética é uma das áreas mais difíceis lidar, de todas as consideradas nas políticas energéticas. Demonstrou-se que muitos dos equipamentos não estão otimizados em termos de gestão de energia, procurou-se evidenciar as consequências dessa realidade, uma vez que os equipamentos referidos, têm a necessidade de estar permanentemente a ser alimentados pela rede de energia elétrica, para garantir as funções para que foram projetados. Da pesquisa efetuada e descrita ao longo da dissertação, constatamos o empenho de toda a comunidade científica, operadores e agências de energia e de telecomunicações, em resolver o problema, já que há a consciencialização de que o ritmo de crescimento da rede e equipamentos terminais, é superior ao registado na melhoria da eficiência energética dos vários componentes e equipamentos terminais. Na segunda parte do relatório da tese, procurou-se testar a aplicabilidade das normas e recomendações dos organismos que tutelam a atividade a nível global - algumas publicadas nos últimos 2 anos - a um caso prático. Um edifício hospitalar de média dimensão. Foi elaborada uma aplicação informática, que suportada numa metodologia padronizada, seja capaz de fazer a avaliação da eficiência energética dos equipamentos serviços de telecomunicações de informação e comunicação em funcionamento do hospital. Por dificuldades de disponibilidade dos responsáveis do edifício, os resultados ficaram aquém do esperado. Conseguiu-se desenhar a aplicação, inventariar-se apenas parte dos equipamentos. Demonstrou-se que, a forma como alguns equipamentos estão a ser utilizados, não cumprem regras de utilização racional e eficiente. Procurou-se sensibilizar alguns dos responsáveis, para a necessidade de alterar comportamentos e prosseguir o processo de inventariação, por forma, a que o trabalho iniciado atinja os objetivos propostos.

MOLECULAR TYPING OF Candida albicans ISOLATES FROM HOSPITALIZED PATIENTS

SUMMARYIntroduction: The majority of nosocomial fungal infections are caused by Candida spp. where C. albicans is the species most commonly identified. Molecular methods are important tools for assessing the origin of the yeasts isolated in hospitals.Methods: This is a study on the genetic profifiles of 39 nosocomial clinical isolates of C. albicans using two typing methods: random amplifified polymorphic DNA (RAPD) and microsatellite, two different primers for each technique were used.Results: RAPD provided 10 and 11 different profiles with values for SAB of 0.84 ± 0.126 and 0.88 ± 0.08 for primers M2 and P4, respectively. Microsatellite using two markers, CDC3 and HIS3, allowed the observation of six and seven different alleles, respectively, with combined discriminatory power of 0.91.Conclusions: Although genetic variability is clear, it was possible to identify high similarity, suggesting a common origin for at least a part of isolates. It is important to emphasize that common origin was proven from yeasts isolated from colonization (urine, catheter or endotracheal secretions) and blood culture from the same patient, indicating that the candidemia must have started from a site of colonization. The combination of RAPD and microsatellite provides a quick and efficient analysis for investigation of similarity among nosocomial isolates of C. albicans.

Segmentectomia Pulmonar Anatómica por Videotoracoscopia: Experiência Inicial de um Centro

The use of minimally invasive surgery for the treatment of lung cancer has been growing worldwide. Between May 2008 and November 2012, we performed 24 videothoracoscopic anatomical lung resections in our department. This includes 22 lobectomies and 2 anatomic segmentectomies, which is known to be a more complex surgery, since it demands a finer dissection of sub-lobar structures. We report the clinical cases of two patients who underwent anatomic segmentectomies. The first one was a 63 year old woman, smoker and with a history of breast cancer 20 years earlier. An incidental 9 mm node was found in the lingula. The patient underwent an anatomic lingulectomy and the frozen section was suggestive of a primary lung cancer. Therefore, we proceeded to a full lymphadenectomy. The final pathology evaluation showed a typical carcinoid tumour (pT1aN0). The second patient was a 50 year old woman, a smoker and with a heavy family history of lung cancer. In a screening CT scan a 8 mm ground glass opacity was identified in the left lower lobe (segment VI). After a VATS wedge resection of the node the frozen section evaluation was compatible with adenocarcinoma. We then proceeded to an anatomic segmentectomy with lymphadenectomy. The definitive pathology evaluation confirmed that it was a pT1a N0 bronchioloalveolar adenocarcinoma. The patients now have 5 and 2 months of follow up respectivelly and neither of them has signs of recurrence and the surgical incision showed a good aesthetic result. Anatomic segmentectomy is the indicated surgery especially in patients with low grade tumours, in early stage lung cancers or in patients without pulmonary function for a lobar resection, and it can be done safely using VATS.

Tentativa de orquidectomia parcial: a propósito de um caso clínico

Introduc¸ão: Até ao fim dos anos 80 defendia-se que qualquer nódulo testicular suspeito devia ser excisado com orquidectomia radical. No entanto, com o aumento do diagnóstico incidental de massas testiculares, a maior acuidade dos exames extemporâneos e a evidência das vantagens potenciais da orquidectomia parcial, questionou-se se seria necessário sacrificar, sempre, todo o testículo, mesmo na presenc¸a de um testículo contralateral normal. Caso clínico: Apresentamos o caso de um doente de 23 anos, com o diagnóstico de um nódulo testicular com 7,5 mm, não palpável, assintomático e marcadores tumorais negativos. Foi submetido a orquidectomia parcial guiada por ecografia e exame extemporâneo, no entanto, por suspeita anatomopatológica de provável tumor de células germinativas, optou-se pela totalizac¸ão da orquidectomia. O resultado histológico final foi de tumor de células de Leydig. Tendo em conta a elevada probabilidade de lesões testiculares não palpáveis e de pequenas dimensões serem benignas (até 80%), os efeitos da orquidectomia radical na espermatogénese, func¸ão endócrina e estética e que não devem ser ignorados, a orquidectomia parcial é um procedimento que, embora não seja um procedimento padrão, pode ser equacionado como primeira abordagem em casos selecionados e em centros de referência especializados.

C-reactive protein at 24 hours after hospital admission may have relevant prognostic accuracy in acute pancreatitis: a retrospective cohort study

Introduction: C-reactive protein (CRP) and Bedside Index for Severity in Acute Pancreatitis (BISAP) have been used in early risk assessment of patients with AP. Objectives: We evaluated prognostic accuracy of CRP at 24 hours after hospital admission (CRP24) for in-hospital mortality (IM) in AP individually and with BISAP. Materials and Methods: This retrospective cohort study included 134 patients with AP from a Portuguese hospital in 2009---2010. Prognostic accuracy assessment used area under receiver---operating characteristic curve (AUC), continuous net reclassification improvement (NRI), and integrated discrimination improvement (IDI). Results: Thirteen percent of patients had severe AP, 26% developed pancreatic necrosis, and 7% died during index hospital stay. AUCs for CRP24 and BISAP individually were 0.80 (95% confidence interval (CI) 0.65---0.95) and 0.77 (95% CI 0.59---0.95), respectively. No patients with CRP24 <60 mg/l died (P = 0.027; negative predictive value 100% (95% CI 92.3---100%)). AUC for BISAP plus CRP24 was 0.81 (95% CI 0.65---0.97). Change in NRI nonevents (42.4%; 95% CI, 24.9---59.9%) resulted in positive overall NRI (31.3%; 95% CI, − 36.4% to 98.9%), but IDI nonevents was negligible (0.004; 95% CI, − 0.007 to 0.014). Conclusions: CRP24 revealed good prognostic accuracy for IM in AP; its main role may be the selection of lowest risk patients.

Magnesium – association with inflammation and renal disease in systemic lupus erythematosus

Introduction: Recent studies suggest that magnesium deficiency may play a role in inflammation. In diabetes and cardio-vascular diseases, conditions with a component of chronic inflammation, C–reactive protein levels are higher and associated with low serum magnesium. The objective of this study is to evaluate serum magnesium levels in patients with systemic lupus erythematosus and its potential association with inflammation and renal manifestations. Methods: All patients with systemic lupus erythematosus followed in a Systemic Immune Diseases Unit, from January 2012 until January 2014, were included in this cross sectional analysis. Patients with infection, neoplasia, liver failure and chronic kidney disease (stage > 3) were excluded. Clinical information and laboratory results (serum magnesium, C-reactive protein, erythrocyte sedimentation rate, serum creatinine and spot urine test) were collected. A multivariate analysis was performed to explore possible predictive factors for hypomagnesaemia. Results: One hundred and two patients were included (94.1% female, 21-86 years). 33.4% had hypertension, 8.8% had diabetes and 20.6% had hypomagnesaemia (< 1.8mg/dL). There were no significant differences between the inflammatory parameters of patients with hypomagnesaemia or normomagnesaemia. Serum magnesium was significantly lower with increasing comorbidities (p = 0.01). Leukocyturia was significantly higher in the hypomagnesaemia group (p = 0.03) and haematuria had a negative correlation with serum magnesium (-0.23, p < 0.05). Multivariate analysis showed that patients with hypertension and diabetes had higher risk of hypomagnesaemia: OR 42.29 (95% CI, 1.43-1249.31). Leukocyturia was also individually and independently associated with hypomagnesaemia: OR 8.37 (95% CI, 1.40-49.97). Conclusion: The presence of hypomagnesaemia in our patients with systemic lupus erythematosus was high. There was no association between the levels of serum magnesium and the inflammatory parameters. Increasing comorbidities and leukocyturia were independent predictors of lower serum magnesium. Finally, the association of leukocyturia and haematuria with lower serum magnesium may suggest a relationship with a higher disease activity.

CORAL SNAKE ANTIVENOM PRODUCED IN CHICKENS (Gallus domesticus)

The production of anti-snake venom from large mammal's blood has been found to be low-yielding and arduous, consequently, antivenom immunoglobulins for treatment are achieved regularly as polyvalent serum. We have standardized an undemanding technique for making purified immunoglobulin IgY antivenom consisting of polyclonal antibodies against coral snake venom in the egg yolk of immunized hens. We have adapted a reported process of antibody purification from egg yolks, and achieved 90% antibody purity. The customized technique consisted of the removal of lipids from distilled water-diluted egg yolks by a freeze–thaw sequence. The specific immunoglobulins were present in the egg yolk for up to 180 days postimmunization. Therefore, by means of small venom quantities, a significant amount of immunoglobulins were found in an adequately purified state (The obtained material contained about 90% pure IgY). The antigen binding of the immunoglobulins was detected by a double immunodiffusion test. Titers of antibodies in the yolk were estimated with a serum protection assay (Median effective dose = ED50) (ED50= 477 mg/kg). Given that breeding hens is economically feasible, egg gathering is noninvasive and the purification of IgY antibodies is quick and easy, chicken immunization is an excellent alternative for the production of polyclonal antibodies. To the best of our knowledge, this is the first coral snake antivenom prepared in birds.

Algorithms for information extraction and signal annotation on long-term biosignals using clustering techniques

Dissertação para obtenção do Grau de Mestre em Engenharia Biomédica

PARTICIPATION OF TICKS IN THE INFECTIOUS CYCLE OF CANINE VISCERAL LEISHMANIASIS, IN TERESINA, PIAUÍ, BRAZIL

In this study, we detected Leishmania spp. infection in R. sanguineus collected from dogs that were naturally infected with L. (L.) infantum. We examined 35 dogs of both sexes and unknown ages. The infected dogs were serologically positive by the immunofluorescence antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA), and Quick Test-DPP (Dual Path Platform), as well as parasitological examination of a positive skin biopsy or sternal bone marrow aspiration. Ten negative dogs were included as controls. The ticks that infested these dogs were collected in pools of 10 adult females per animal. The PCR was performed with specific primers for Leishmania spp., which amplified a 720-bp fragment. Of the 35 analyzed samples, a product was observed in eight samples (8/35; 22.9%). We conclude that the presence of parasite DNA suggests that ticks participate in the zoonotic cycle of canine visceral leishmaniasis, in the city of Teresina, Piauí.

LABORATORY EVALUATION OF THE DEVELOPMENT OF Aedes aegypti IN TWO SEASONS: INFLUENCE OF DIFFERENT PLACES AND DIFFERENT DENSITIES

Aedes aegypti is an important vector in Brazil being the main vector of the dengue-fever. This paper employs survival curves to describe the time in days from larvae to adult forms of Aedes aegypti raised, individually and collectively, and compares it during winter and spring when positioned inside and outside a laboratory. The study was conducted in São Vicente, a coastal city in Southeastern Brazil. The lowest water temperature in winter and in spring was 20 °C and the highest was 26 °C in spring. Higher and more stable temperatures were measured in the intra compared to the peri in both seasons. Consequently, larvae positioned in the intra resulted in the lowest median time to develop in the individual and collective experiment (nine and ten days, respectively). At least 25% of the larvae positioned in the intra in the individual experiment in the spring took only seven days to reach adulthood. Sex ratios and the median time development by sex did not show significant differences. These results indicate that efforts to control Aedes aegypti must be continuous and directed mainly to prevent the intra-domiciliary sites that can be infested in a week in order to reduce the human-vector contact.