HIGH SENSITIVE PCR METHOD FOR DETECTION OF PATHOGENIC Leptospira spp. IN PARAFFIN-EMBEDDED TISSUES

This study describes the development and application of a new PCR assay for the specific detection of pathogenic leptospires and its comparison with a previously reported PCR protocol. New primers were designed for PCR optimization and evaluation in artificially-infected paraffin-embedded tissues. PCR was then applied to post-mortem, paraffin-embedded samples, followed by amplicon sequencing. The PCR was more efficient than the reported protocol, allowing the amplification of expected DNA fragment from the artificially infected samples and from 44% of the post-mortem samples. The sequences of PCR amplicons from different patients showed >99% homology with pathogenic leptospires DNA sequences. The applicability of a highly sensitive and specific tool to screen histological specimens for the detection of pathogenic Leptospira spp. would facilitate a better assessment of the prevalence and epidemiology of leptospirosis, which constitutes a health problem in many countries.

FALSE-NEGATIVE DENGUE CASES IN RORAIMA, BRAZIL: AN APPROACH REGARDING THE HIGH NUMBER OF NEGATIVE RESULTS BY NS1 AG KITS

Serum samples from 150 NS1-negative (Platelia ELISA) patients presumptively diagnosed with dengue were analyzed by the TaqMan probed real-time reverse transcription PCR (TaqMan qRT-PCR) method. The qRT-PCR positive samples were tested for serotype by semi-nested RT-PCR and a qualitative immunochromatographic assay for IgG and IgM. Molecular detection methods showed 33 (22%) positive samples out of 150 NS1-antigen negative samples. Of these, 72% were collected up to day 2 after the onset of symptoms, when diagnostic sensitivity of NS1-antigen test assays is significantly enhanced. Most of the cases were not characterized as secondary infection. Twenty-eight samples were successfully serotyped, 75% of which for DENV-4, 14% for DENV-2, 7% for DENV-3 and 4% for DENV-1. These findings reaffirm the hyperendemic situation of the state of Roraima and suggest a lower sensitivity of the NS1 test, mainly when DENV-4 is the predominant serotype. Health care providers should therefore be aware of samples tested negative by NS1 antigen assays, especially when clinical symptoms and other laboratory data results show evidence of dengue infection.

Ponseti Method: Does Age at the Beginning of Treatment Make a Difference?

The Ponseti method is reportedly effective for treating clubfoot in children up to 9 years of age. However, whether age at the beginning of treatment influences the rate of successful correction and the rate of relapse is unknown. We therefore retrospectively reviewed 68 consecutive children with 102 idiopathic clubfeet treated by the Ponseti technique in four Portuguese hospitals. We followed patients a minimum of 30 months (mean, 41.4 months; range, 30–61 months). The patients were divided into two groups according to their age at the beginning of treatment; Group I was younger than 6 months and Group II was older than 6 months. All feet(100%) were initially corrected and no feet required extensive surgery regardless of age at the beginning of treatment. There were no differences between Groups I and II in the number of casts, tenotomies, success in terms of rate of initial correction, rate of recurrence, and rate of tibialis anterior transference. The rate of the Ponseti method in avoiding extensive surgery was 100% in Groups I and II; relapses occurred in 8% of the feet in younger and older children. Level of Evidence: Level II, prognostic study. See the Guidelines for Authors for a complete description of levels of evidence.

The use of long acting sulfonamides, alone or with pyrimethamine, in malaria (with special reference to sulformetoxine)

Review of the early literature as well as more recent results show that sulfonamides possess a distinct antimalarial activity. However, when give alone, their action is less marked and slower than that of the antimalarials commonly used in the treatment of the acute attack. Combinations with pyrimethamine provide better results, even in cases of pyrimethamine and chloroquine resistance. This warrants further investigations in an attempt to develop a therapeutic agent suitable for the treatment of such resistant cases. It may also be possible with an appropriate combination of pyrimethamine with a sulfonamide to achieve a satisfactory method for suppressive treatment both in areas with and without pyrimethamine resistance. Three main points must still be carefully studied: 1) the risk of developing malaria resistance against one or both of the components of the combination. 2) The risk of developing bacterial resistance to sulfonamides if these substances are used on a large scale in too low doses. It seems indeed that antimalarial effect with the combination of sufonamides + pyrimethamine can be obtained with doses of sulfonamides which are below those usually employed in bacterial diseases. Since the range of the ratios providing potentiation is rather large, only ratios of the combination sulfonamides: pyrimethamine should be chosen in which an antfbacterial sulfonamidemia is guaranteed. 3) It goes without sayinq that, although both pyrimethamine and modem sulfonamides, when given by themselves, have proved tc possess a large margin of safety, long term administration of their combination should be careful studied from the point of view of possible side effects. Substantial evidence has already been produced to show that the long acting sulfonamide Fanasil (Ro 4-4393) given once or once weekly possesses marked schizonticidal activity against P. falciparum. Although its action is slower than that of 4-aminoquinolines, it may be useful as a second choice drug in semi-immune subjects for the therapy of falciparum malaria. Preliminary results show that, when combined with pyrimethamine, Fanasil is highly effective in suppressing fever and asexual parasitemia due to P. falciparum. Single doses of 1 g Fanasil together with 50 mg pyrimethamine seem to be adequate for the treatment of acute falciparum malaria in semi-immune patients. The onset of action of the combination is much more rapid than that of the single components. Weekly doses of 500 mg Fanasil and 25 mg pyrimeihamine appear to provide satisfactory suppressive effects against P. falciparum at least in East Africa. This combination is active on strains which do not respond satisfactorily to the standard doses of pyrimethamine and/or chloroquine and seems to have a satisfactory sporontocidal effect. Preliminary results indicate that Fanasil alone cannot be recommended for use against the other human malaria parasites. The combination with pyrimethamine appears to be much more effective. East African strains of P. malariae seem to respond better to the combination than do Malayan strains of P. vivax but further trials are required before definite assessment can be made. Fanasil by itself has no gametocytoddal or sporontocidal action but seems to potentiate the effect of pyrimethamine at least on sporogony of P. falciparum.

O estudo de pavimentações regulares e semi-regulares com ambiente de geometria dinâmica

Dissertação para obtenção do Grau de Mestre em Ensino da Matemática

Avaliação sorológica da vacina oral, tipo Sabin, contra a poliomielite em região semi-rural: I. Formação de anticorpos em vacinados

Os autores descrevem, os resultados obtidos em um programa de avaliação sorológica da vacina oral, tipo Sabin, contra a poliomielite, em uma comunidade semi-rural, próxima a cidade do Rio de Janeiro. Em condições controladas 114 crianças, com idades entre 3 meses a 3 anos (Tabela 1) foram vacinadas, com vacinas trivalentes (500.000, 200.000, e 300.000 TCD50 por dose, dos tipos 1, 2 e 3 respectivamente), usando-se três doses, com intervalos de 8 semanas entre as doses. Amostras de sangue foram coletadas por punção venosa ou discos de papel de filtro, juntamente com a 1.ª e a 3.ª dose de vacina e 9 semanas após esta última dose de vacina. As taxas de conversão alcançaram (diluição de sôro 1/8) 82,7%, 98,5% e 75,4% para os tipos 1, 2 e 3 respectivamente, após três doses de vacina (Tabela 2). A distribuição de idade de indivíduos sem anticorpos após a vacinação (Tabela 3) mostra o grupo etário de 1 a 2 anos como o que apresenta a mais baixa taxa de conversão. Os autores acentuam que as condições de vida da população estudada correspondem àquelas de grandes partes da população brasileira, nas áreas rurais do país; e uma avaliação semelhante da vacina em áreas urbanas, seria desejável. Os autores sugerem ainda o aumento da quantidade de vírus do tipo 1 na vacina como medida provàvelmente eficaz na melhora das taxas de conversão em populações como a estudada. Estudos quantitativos sôbre anticorpos para Enterovírus, presentes na população estudada, estão sendo realizados e serão pròximamente apresentados.

Avaliação sorológica da vacina oral, tipo Sabin, contra a poliomielite em região semi - rural: II - Aspectos quantitativos dos anticorpos formados e anticorpos para outros enterovirus

Os autores apresentam dados quantitativos sôbre os anticorpos presentes em vacinados com vacina oral trivalente, contra a poliomielite (Tabela 1). Apos a 3.ª dose de vacina verificou-se um aumento do título geométrico médio da população em relação aos títulos obtidos com duas doses de vacina. Infecções naturais devem ter contribuído para formação de anticorpos para poliomielite na população, ao lado da vacina. Anticorpos para enterovírus não-pólio (Coxsackie B e alguns tipos de vírus ECHO) são apresentados na Tabela II e referem-se a amostras de sôro colhidas quando da 1.ª dose de vacina. Os autores chamam a atenção para a incidência de enterovírus na região, embora poucos sejam os dados ainda disponíveis.

Preparation of gas-filled porous microparticles (GPPs) and microbubbles (MBs) by PGSS method

Dissertation to obtain the Master Degree in Biotechnology

Ciclo Vital de Schistosoma mansoni através do Holochilus brasiliensis (Desmarest, 1818) em ambiente semi-natural (Trematoda, Shistosomatidae; Rodentia, Cricetidae)

Junto ao Lago da Pampulha, Belo Horizonte, MG, foram capturados (julho/72-novembro/73) 28 exemplares de Holochilus brasiliensis, dos quais 11 (39,3%) eliminavam nas fezes ovos viáveis de S. mansoni. Miracídios da cepa mencionada ("H") infectaram Biomphalaria glabrata e as cercárias obtidas também infectaram camundongos albinos, recuperando-se, ao final do experimento, 35,3% de vermes adultos. Por outro lado, cercárias de cepa humana ("LE") de S. mansoni infectaram sete exemplares de H. brasiliensis, nascidos em laboratório, recuperando-se no fim de 60 dias, 30,5% de vermes adultos. Estudos anatomapatológicos de H. brasiliensis demonstraram infecção generalizada, encontrando-se granuloma no esôfago, estômago, intestino (delgado e grosso), fígado, baço, pâncreas e linfonodos abdominais. Espessamentos fibrosos da íntima da veia porta, granulomas em espaços porta e fibrose incipiente dos espaços porta e interlobular foram lesões decorrentes da presença de ovos de S. mansoni encontrados no fígado. Em ambiente semi-natural, foi possível fechar o ciclo do S. mansoni sem direta participação humana, utilizando-se B. glabrata experimentalmente infectadas com trematódeos da cepa "LE", H. brasiliensis nascidos em laboratório e B. glabrata nascida no ambiente semi-natural estabelecido. Verificou-se que ambas as cepas ("H" e "LE") comportaram-se de maneira análoga, não sendo verificadas, também, diferenças morfológicas entre os ovos e vermes adultos de ambas. As observações, realizadas no campo e no laboratório demonstraram que o Holochilus brasiliensis é bom hospedeiro de Schistosoma mansoni. Assim, em determinadas áreas e sob certas condições ecológicas, o cricetídeo em questão poderá, efetivamente, integrar-se ao ciclo do trematódeo na natureza, independente ou paralelamente à presença do homem. Assinala-se, finalmente, que o presente trabalho relata o segundo fechamento do ciclo biológico de S. mansoni em condições ditas semi-naturais. Os primeiros estudos, entretanto, de Antunes, Milward de Andrade, Katz & Coelho4,,em 1971 e de Antunes5, em 1971 foram feitos utilizando-se o Nectomys s. squamipes.

The sum-of-the-parts method: An international application

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

A Homemade Snare: An Alternative Method for Mechanical Removal of Dirofilaria Immitis in Dogs

Canine dirofilariosis is a life-threatening parasitic disease that is increasingly reported worldwide. Once diagnosed the main treatment goals are to improve the animal's clinical condition and to eliminate all life stages of the parasite with minimal posttreatment side effects. This can be achieved through mechanical, surgical, or chemotherapeutical approaches. Currently, manual extraction is the preferred method to remove adult heartworms due to its diminished invasiveness, reduced damage to the vascular endothelium, and shortened anaesthesia duration. However, it remains an expensive technique that can be highly traumatic. To address this issue, a nontraumatic homemade catheter-guided snare was developed for heartworm removal by adapting and folding a 0.014-inch coronary wire (BMW, Abbott Vascular). Transvenous heartworm extraction was performed on a dog severely infected with adult heartworms by inserting the modified snare into a 6-F Judkins right coronary guiding catheter BMW (Cordis) and advancing it into the right ventricle under fluoroscopic guidance. Fifteen adult specimens of Dirofilaria immitis were successfully extracted from the pulmonary artery and right ventricle without complications. To assure the death of both larvae and adults, postoperative treatment was successfully managed using ivermectin, doxycycline, and melarsomine, with no recurrence after surgery.

What is the best accounting method for financial assets?

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

Osmolality of Elemental and Semi-Elemental Formulas Supplemented with Nonprotein Energy Supplements

Elemental and semi-elemental formulas are used to feed infants with short bowel syndrome, who may not be able to tolerate feeds of more than 310 mOsm kg(-1). The present study aimed to measure the osmolality of elemental and semi-elemental formulas at different concentrations, with and without the addition of nonprotein energy supplements.

The influence of burden of care and perceived stigma on expressed emotions of relatives of stable persons with schizophrenia in Nigerian semi-urban/urban settings

RESUMO: Schizophrenia’s burden defines experience of family members and is associated with high level of distress. Courtesy stigma, a distress concept, worsens caregivers’ burden of care and impacts on schizophrenia. Expressed emotion (EE), another family variable, impacts on schizophrenia. However, relationship between EE, burden of care and stigma has been little explored in western literature but not in sub-Saharan Africa particularly Nigeria. This study explored the impact of burden of care and courtesy stigma on EE among caregivers of persons with schizophrenia in urban and semi-urban settings in Nigeria. Fifty caregivers each from semi-urban and urban areas completed a socio-demographic schedule, family questionnaire, burden interview schedule and perceived devaluation and discrimination scale. The caregivers had a mean age of 42 (± 15.6) years. Majority were females (57%), married (49%), from Yoruba ethnic group (68%), monogamous family (73%) and Christians (82%). A higher proportion of the whole sample (53%) had tertiary education. Three out of ten were sole caregivers. Seventy three (73%) lived with the person they cared for. The average number of hours spent per week by a caregiver with a person with schizophrenia was 35 hours. The urban sample had significantly higher proportion of carers with high global expressed emotion (72.7%) than the semi-urban sample (27.3%). The odds of a caregiver in an urban setting exhibiting high expressed emotion are 4.202 times higher than the odds of caregiver in a semi-urban setting. Additionally, there was significance difference between the urban and semi-urban caregivers in discrimination dimension. High levels of subjective and objective burden were associated with high levels of critical comments. In conclusion, this study is the first demonstration of urban-semi-urban difference in expressed emotion in an African country and its findings provide further support to hypothesized relationship between components of EE and burden of care.

Speed control of induction machine based on direct torque control method

Dissertação para obtenção do Grau de Mestre em Engenharia Electrotécnica e de Computadores