SURVIVAL, INDUCTION AND RESUSCITATION OF Vibrio cholerae FROM THE VIABLE BUT NON-CULTURABLE STATE IN THE SOUTHERN CARIBBEAN SEA

The causative agent of cholera, Vibrio cholerae, can enter into a viable but non-culturable (VBNC) state in response to unfavorable conditions. The aim of this study was to evaluate the in situ survival of V. cholerae in an aquatic environment of the Southern Caribbean Sea, and its induction and resuscitation from the VBNC state. V. cholerae non-O1, non-O139 was inoculated into diffusion chambers placed at the Cuare Wildlife Refuge, Venezuela, and monitored for plate, total and viable cells counts. At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state. Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells. Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season. Resuscitation studies showed significant recovery of cell cultivability with spent media addition (p < 0.05). These results suggest that V. cholerae can persist in the VBNC state in this Caribbean environment and revert to a cultivable form under favorable conditions. The VBNC state might represent a critical step in cholera transmission in susceptible areas.

Contribuição ao estudo da mistura néon-azoto a baixas temperaturas

Dissertação para obtenção do Grau de Mestre em Engenharia Física

FIRST REPORT OF ACUTE CHAGAS DISEASE BY VECTOR TRANSMISSION IN RIO DE JANEIRO STATE, BRAZIL

SUMMARY Chagas disease (CD) is an endemic anthropozoonosis from Latin America of which the main means of transmission is the contact of skin lesions or mucosa with the feces of triatomine bugs infected by Trypanosoma cruzi. In this article, we describe the first acute CD case acquired by vector transmission in the Rio de Janeiro State and confirmed by parasitological, serological and PCR tests. The patient presented acute cardiomyopathy and pericardial effusion without cardiac tamponade. Together with fever and malaise, a 3 cm wide erythematous, non-pruritic, papule compatible with a "chagoma" was found on his left wrist. This case report draws attention to the possible transmission of CD by non-domiciled native vectors in non-endemic areas. Therefore, acute CD should be included in the diagnostic workout of febrile diseases and acute myopericarditis in Rio de Janeiro.

HEPATOCELLULAR CARCINOMA IN A NON-CIRRHOTIC PATIENT WITH SUSTAINED VIROLOGICAL RESPONSE AFTER HEPATITIS C TREATMENT

Chronic infection by hepatitis C virus (HCV) is one of the main risk factors for the development of liver cirrhosis and hepatocellular carcinoma. However, the emergence of hepatocellular carcinoma (HCC) in non-cirrhotic HCV patients, especially after sustained virological response (SVR) is an unusual event. Recently, it has been suggested that HCV genotype 3 may have a particular oncogenic mechanism, but the factors involved in these cases as well as the profile of these patients are still not fully understood. Thus, we present the case of a non-cirrhotic fifty-year-old male with HCV infection, genotype 3a, who developed HCC two years after treatment with pegylated-interferon and ribavirin, with SVR, in Brazil.

Non-clinical isolates bring new findings on enterococcal virulence

Dissertation presented to obtain the PhD degree in Biology

FACTORS RELATED TO ORAL CANDIDIASIS IN ELDERLY USERS AND NON-USERS OF REMOVABLE DENTAL PROSTHESES

This study examined the association between oral candidiasis in elderly users and nonusers of prosthesis and its predisposing factors. To this end, we performed a cross-sectional study where saliva samples from 48 patients were collected they used prosthesis and 43 patients (control group) who did not use. Among the 91 patients, Candida spp were isolated in 40 (83.3%) who used prosthesis and in 23 (53.5%) in the control group. A statistically significant association was determined between the two groups, the isolation of yeasts and dental prosthesis (p < 0.05, OR = 4.3). The most common etiological agent was Candida albicans (37 isolates), with 23 (62.2%) in the denture group and 14 (37.8%) (control group). Among patients who presented clinical manifestations of oral candidiasis (n = 24), 83.3% (n = 20) belonged to the group that wore dentures, while only 16.7% (n = 4) belonged to the control group. Elderly patients with diabetes had 4.4 times higher estimated risk of developing oral candidiasis when compared with individuals without this condition. There was no statistically significant association between being user prostheses and have diabetes with the onset of candidiasis. No statistically significant association was determined between xerostomia, use of prosthesis and oral candidiasis. The use of prosthetics and poor oral hygiene in elderly patients predisposes to the development of oral candidiasis.

Atheroembolic Renal Disease As a Cause of Allograft Primary Non-Function

Atheroembolic renal disease, also referred to as cholesterol crystal embolization, is a rare cause of renal failure, secondary to occlusion of renal arteries, renal arterioles and glomerular capillaries with cholesterol crystals, originating from atheromatous plaques of the aorta and other major arteries. This disease can occur very rarely in kidney allografts in an early or a late clinical form. Renal biopsy seems to be a reliable diagnostic test and cholesterol clefts are the pathognomonic finding. However, the renal biopsy has some limitations as the typical lesion is focal and can be easily missed in a biopsy fragment. The clinical course of these patients varies from complete recovery of the renal function to permanent graft loss. Statins, acetylsalicyclic acid, and corticosteroids have been used to improve the prognosis. We report a case of primary allograft dysfunction caused by an early and massive atheroembolic renal disease. Distinctive histology is presented in several consecutive biopsies. We evaluated all the cases of our Unit and briefly reviewed the literature. Atheroembolic renal disease is a rare cause of allograft primary non -function but may become more prevalent as acceptance of aged donors and recipients for transplantation has become more frequent.

Evolutive pattern of schistosomiasis and life-span of S. mansoni in patients living in non-endemic area in Brazil

Out of 2484 patients harboring S. mansoni seen in Rio de Janeiro, 1197 had been living permanently out of endemic area frorn one to 30 years, without any possibility of reinfection; 90.1% of these 1197 patients were first seen with, hepato-intestinal schistosomiasis and only 9.9% with hepatosplenic form. 55% of thern still had S. mansoni active infection 6 years or more after they had left the endemic area and 26.5% remained infected for more than 10 years. The patients with intestinal or hepato-intestinal schistosomiasis did not develop the most severe form whether they had been treated or not, and the hepatosplenic patients had a long time to deteriorate.

Non-Progressive Leukoencephalopathy with Bilateral Temporal Cysts

We report two cases of a peculiar leukoencephalopathy with temporal cysts. Both patients have a non-progressive neurological disorder with mental retardation, microcephaly and sensorineural deafness although clinical differences between them may reflect a different aetiology. The metabolic disorders with white matter involvement and the recently described leukoencephalopathies (Van Der Knaap disease, 'vanishing white matter disease') were excluded based on clinical, biologic and imaging findings. Cytomegalovirus infection is a likely possibility in the first case although the magnetic resonance imaging picture is only partially similar to previously reported cases. Our patients are strikingly similar to the patients reported by Deonna et al. and Olivier et al. We discuss the clinical and imaging findings in our patients and the differential diagnosis considering the known disorders of the white matter in childhood.

Cost-Effectiveness of Non-Vitamin K Antagonist Oral Anticoagulants for Atrial Fibrillation in Portugal

INTRODUCTION AND OBJECTIVES:Recently, three novel non-vitamin K antagonist oral anticoagulants received approval for reimbursement in Portugal for patients with non-valvular atrial fibrillation (AF). It is therefore important to evaluate the relative cost-effectiveness of these new oral anticoagulants in Portuguese AF patients. METHODS: A Markov model was used to analyze disease progression over a lifetime horizon. Relative efficacy data for stroke (ischemic and hemorrhagic), bleeding (intracranial, other major bleeding and clinically relevant non-major bleeding), myocardial infarction and treatment discontinuation were obtained by pairwise indirect comparisons between apixaban, dabigatran and rivaroxaban using warfarin as a common comparator. Data on resource use were obtained from the database of diagnosis-related groups and an expert panel. Model outputs included life years gained, quality-adjusted life years (QALYs), direct healthcare costs and incremental cost-effectiveness ratios (ICERs). RESULTS:Apixaban provided the most life years gained and QALYs. The ICERs of apixaban compared to warfarin and dabigatran were €5529/QALY and €9163/QALY, respectively. Apixaban was dominant over rivaroxaban (greater health gains and lower costs). The results were robust over a wide range of inputs in sensitivity analyses. Apixaban had a 70% probability of being cost-effective (at a threshold of €20 000/QALY) compared to all the other therapeutic options. CONCLUSIONS:Apixaban is a cost-effective alternative to warfarin and dabigatran and is dominant over rivaroxaban in AF patients from the perspective of the Portuguese national healthcare system. These conclusions are based on indirect comparisons, but despite this limitation, the information is useful for healthcare decision-makers.

Effects of lethal and non-lethal malaria on the mononuclear phagocyte system

The effects ofone non-lethal species ofmalarialparasite, Plasmodium yoelii, and one lethal species, P. berghei, on the mononuclear phagocyte system (MPS) of BALB/c mice were studied. P. yoelii caused a greater and more sustained expansion and activation of the MPS, and the two major populations of spleen phagocytic cells-red pulp and marginal zone macrophages - exhibited a greater increase in numbers in this infection. During the course of P. berghei mataria, the spleen was progressively occupied by haematopoietic tissue and, at the terminal stage of infection, an extensive depletion of lymphocytes and macrophages was apparent. The possibility was suggested that the outcome of mataria may be inftuenced by the particular way the parasite interacts with the MPS.

Infecção por Treponema pallidum: análise serológica e pesquisa de DNA

RESUMO: A sífilis é uma infecção causada por T. pallidum que pode ser transmitida por via vertical, por contacto sexual ou por sangue, e cujo diagnóstico se baseia na associação entre manifestações clinicas e testes serológicos. Neste estudo utilizaram-se os testes serológicos RPR, TPHA, FTA-Abs, um teste rápido não comercializado (Signal-Spirolipin) que pesquisa anticorpos treponémicos (CDC-T) e não treponémicos (CDC-2) em simultâneo no mesmo dispositivo e uma técnica de PCR-multiplex para a pesquisa de ADN de T. pallidum. Da comparação das técnicas serológicas constatou-se que a sensibilidade dos testes RPR e TPHA foi de 84,5% e 98% respectivamente. A especificidade foi de 77,3% para o teste RPR e de 87,5% para o teste TPHA. Na avaliação do teste rápido a comparação do teste CDC-2 com o teste RPR resultou numa taxa de concordância de 93,1% enquanto que, a do teste CDC-T com o teste TPHA foi de 94,8%. A sensibilidade e especificidade obtidas pelos testes CDC-2 e CDC-T foram de 88,7% e 65% e de 94,9% e 91,3% respectivamente. Considerando o diagnóstico de sífilis activa com base na reactividade simultânea dos testes RPR e TPHA, avaliou-se a sensibilidade do teste rápido utilizando esse mesmo critério. A sensibilidade obtida foi de 98,4% para o teste rápido e de 97,2% para a associação dos testes RPR/TPHA. A técnica de PCR-multiplex apresentou fraca sensibilidade já que em apenas 33% dos casos de sífilis foi identificado ADN de T. pallidum. O teste rápido avaliado apresentou um comportamento idêntico aos testes geralmente utilizados na rotina laboratorial tais como os utilizados neste estudo. Apresenta a vantagem de pesquisar anticorpos treponémicos e não treponémicos, ao contrário dos testes rápidos comercializados que pesquisam apenas anticorpos treponémicos. Não sendo necessário para a sua execução equipamento laboratorial especializado poderá ser de grande utilidade para o clinico a exercer em locais sem laboratório.------------- ABSTRACT: Syphilis is an infection caused by T. pallidum. Transmission may be vertical, through sexual contact or blood. The diagnosis is based in both serology and clinical manifestations. In this study, we used the serological tests RPR, TPHA, FTA-Abs and a non -commercialized rapid test (Signal-Spirolipin) to detect both treponemal (CDC-T) and non – treponemal antibodies (CDC-2) in the same device. T. pallidum DNA was identified with a multiplex PCR technique. In this study, the RPR and TPHA tests sensitivity was 84,5% and 98%, respectively, and the specificity 77,3% for the RPR test and 87,5% for TPHA test, respectively. The concordance rate was 93,1% in the comparison between the RPR and the CDC2 tests and 94,8% between the CDC-T with the TPHA tests. Sensitivity and specificity of the CDC-2 and CDC-T tests were 88,7% and 65% and 94,9% e 91,3%, respectively. Taking into account that the diagnosis of active syphilis is made when both the RPR and TPHA tests are reactive, we evaluated the sensitivity of the rapid test (CDC2 and CDCT) in the diagnosis of active syphilis, using the above described criteria. The sensitivity was 98,4 % for the rapid test and 97,2 % for the association of reactivity in both RPA and TPHA tests. The multiplex PCR technique presented a low sensitivity, with T. pallidum DNA being identified in only 33% of the syphilis cases. The rapid test evaluated in this study presented identical results to the tests generally used in the routine laboratory diagnosis, such as those used here. However, it does have the advantage of detecting both treponemal and non – treponemal antibodies what is not the case of the commercialized rapid test. Furthermore, there is no need of specialized laboratory equipment, which makes it of great utility in regions where such conditions do not exist, as in developing countries.

How to sell successfully a perfume in the non-selective market?

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA – School of Business and Economics

Identity dynamics in non-profit organizations: Ideological and methodological identity orientations

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA – School of Business and Economics

How differently do our non-large companies listed on the Portuguese Stock Exchange perform?

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Economics from the NOVA – School of Business and Economics