DOT-ELISA for detection of anti-Cysticercus cellulosae antibodies in human cerebrospinal fluid using a new solid-phase (resin-treated polyester fabrics): preliminary report

A dot enzyme-linked immunosorbent assay (DOT-ELISA) was developed to detect specific antibodies in cerebrospinal fluid (CSF) for human neurocysticercosis immunodiagnosis, with Cysticercus cellulosae antigen dotted on a new solid-phase. This was represented by sheets of a synthetic polyester fabric impregnated with a polymerized resin (N-methylol-acrylamide). A very stable preparation was thus obtained, the antigen being covalently bound by cross-linking with free N-methylol groups on the resin. Since robust, no special care was necessary for handling the solid-phase. The test could be performed at room-temperature. From 30 CSF samples assayed, 14 were positive, from a group of 15 cases of neurocysticercosis, with titers from 1 to 128; 15 other samples, from normals or other neurological diseases, were all negative. Test characteristics seem to indicate it as adequate for epidemiological surveys. A more detailed study on sensitivity, specificity, reproducibility and the use in serum samples is being conducted.

UltramicroELISA para la detección de anticuerpos IgM anti M. leprae

La disponibilidad del sistema Ultramicroanalítico (SUMA) y de un antígeno especie-específico del M. leprae obtenido mediante síntesis química, permitió la normalización y validación de un ultramicroELISA para la detección de anticuerpos IgM específicos a esta micobacteria. El análisis de 433 sueros de banco de sangre y 265 sueros usados para validar el método y clasificados en un grupo control de donantes de banco de sangre (100), un grupo de pacientes tuberculosos (50), un grupo de enfermos de lepra (65) y un grupo de contactos de estos enfermos (50), mostró la especificidad del ensayo para evidenciar la infección con el M. leprae. Los resultados obtenidos del estudio adicional de 140 muestras de suero de contactos de enfermos estuvieron estrechamente correlacionados (r = 0,98) con los resultados obtenidos por la técnica de microELISA convencional. La utilización del SUMA no solo permite un notable ahorro de reactivos si no además facilita la lectura, cálculo, validación y almacenamiento automático de los resultados.

Pre-meal tomato (Lycopersicon esculentum) intake can have anti-obesity effects in young women?

The effect of pre-meal tomato intake in the anthropometric indices and blood levels of triglycerides, cholesterol, glucose, and uric acid of a young women population (n=35, 19.6 ± 1.3 years) was evaluated. During 4 weeks, daily, participants ingested a raw ripe tomato (~90 g) before lunch. Their anthropometric and biochemical parameters were measured repeatedly during the follow-up time. At the end of the 4 weeks, significant reductions were observed on body weight (-1.09 ± 0.12 kg on average), % fat (-1.54 ± 0.52%), fasting blood glucose (-5.29 ± 0.80 mg/dl), triglycerides (-8.31 ± 1.34 mg/dl), cholesterol (-10.17 ± 1.21 mg/ dl), and uric acid (-0.16 ± 0.04 mg/dl) of the participants. The tomato pre-meal ingestion seemed to interfere positively in body weight, fat percentage, and blood levels of glucose, triglycerides, cholesterol, and uric acid of the young adult women that participated in this study.

Prevalências do HBsAg, do anti-HBc e do anti-HCV na população de candidatos a doadores de sangue do Hemocentro-Campinas

Em 29833 doadores pesquisados encontramos prevalência de 1,52% para o HBsAg e de 11% para o anti-HBc. A co-positividade anti-HBc/anti-HBs em 2783 doadores HBsAg negativos/anti-HBc positivos foi de 81,9%. A prevalência para o HBsAg é baixa nos doadores de Campinas, enquanto o anti-HBc apresenta-se com prevalência elevada quando comparado a outros países. A pesquisa do anti-HCV, em doadores de sangue de Campinas, mostrou prevalência de 2,6% para este marcador, que é bem maior que as observadas nos EUA e Europa. Cerca de 36% dos doadores anti-HCV positivos são anti-HBc reagentes, permitindo inferir, que estas duas viroses acometem simultânea ou sequencialmente os doadores de sangue brasileiros.

Hepatites pós-transfusionais na cidade de Campinas, SP, Brasil: II. Presença dos anticorpos anti-HBc e anti-HCV em candidatos a doadores de sangue e ocorrência de hepatites pós-transfusionais pelo vírus C nos receptores de sangue ou derivados

Pesquisamos os anticorpos anti-HBc e anti-HCV em amostras de soros provenientes de 799 candidatos a doadores, que tiveram suas unidades de sangue ou derivados transfundidas a 111 receptores. O anti-HBc e o anti-HCV foram reagentes, em respectivamente 9 e 2,1% dos doadores testados. Observamos que entre os 111 receptores, 44 haviam recebido pelo menos uma unidade anti-HBc positiva e 67 haviam sido transfundidos somente com unidades anti-HBc negativas. Houve um risco 4,5 vezes maior de aquisição de hepatite por vírus C pelos receptores que receberam pelo menos uma unidade anti-HBc positiva Se a pesquisa do anti-HBc fosse realizada na triagem sorológica dos doadores de sangue, cerca de 56% dos casos de HVC nos receptores saiam evitados. A população de receptores que recebeu pelo menos uma unidade anti-HCV reagente, apresentou um risco 29 vezes maior de adquirir esta hepatite, quando comparada aos receptores transfundidos com todas as unidades anti-HCV negativas. A realização do teste para a pesquisa do anti-HCV na triagem dos doadores de sangue, preveniria 79% dos casos de HVC pós-transfusionais. Os candidatos a doadores brasileiros parecem ser acometidos simultânea ou sequencialmente, pelos vírus B e C das hepatites, pois, 44,4% dos doadores anti-HCV positivos, também foram anti-HBc positivos. A realização dos testes para as pesquisas dos anticorpos anti-HBc e anti-HCV, nas triagens hemoterápicas, está indicada para prevenir a transmissão de hepatites pós-transfusionais, em nosso meio.

Reduced schedule of human anti-rabies immunization with Fuenzalida & Palacios vaccine: additional data

It was reevaluated a reduced schedule for anti-rabies post-exposure immunization with newborn mice nervous tissue vaccine (Fuenzalida 8c Palacios) in a group of 30 non exposed volunteers. The vaccine was administered by intramuscular injections on days zero, 2, 4, 16 and 27, in the deltoid area. Antibody levels were determinated by a simplified serum neutralization microtest on days zero, 16 and 37. On days 16 and 37 the antibody levels of the whole group was >0.5 IU/ml and >1.0 IU/ml, respectively. The cell mediated immunity was precociously detected (on day 4) by the delayed type hipersensitivity skin test. Our results show that this reduced schedule elicited an early and effective humoral and cellular immune response. However it is necessary other studies with larger groups of vaccinees in order to obtain definitive conclusion.

Detection of hepatitis B virus DNA by the polymerase chain reaction in anti-HBE positive chronic hepatitis B patients

Detection of HBV-DNA by PCR was compared with other serological markers (HBsAg, HBeAg and anti-HBe) in a series of49 Chronic Hepatitis B patients, including 12 with a spontaneous clearance of HBsAg. None of these HBsAg negative cases were PCR positive, but 33/37 (89.2%) HBsAg positive cases were PCR positive (p < 0.0001). Among HBsAg positive samples, nine cases were HBeAg positive and anti-HBe negative, all of them PCR positive. Other 3 patients were HBeAg and anti-HBe positive and these cases were also found PCR positive. A third group included 21 patients anti-HBe positive and HBeAg negative: 19 of them were PCR positive and 2 were PCR negative. The last 4 cases were HBeAg and anti-HBe negative, two of them were PCR positive. The detection of anti-HBe viremic cases in the present series suggest that preC variants could occur in our country. In conclusion, the integrated phase o f chronic hepatitis B seems to be less frequent than it was assumed, when only HBeAg or dot blot hybridization techniques were used. The new term "low replication phase" might favorably replace the former "integrated phase".

Seasonal variation of anti-resa/Pf155 Plasmodium falciparum antibodies in three localities from the state of Amapá, Brazil

Anti-RESA/Pf155 antibodies were assayed in sera of individuals from three localities (Laranjal do Jari, Vila Padaria and Vila Paraíso) in the State of Amapá, Brazil, during the long-rains and short-rains seasons. All of these had negative blood smears for malaria. Most of the sera collected were positive in Indirect Fluorescent Antibody (IFA) with P. falciparum parasites, with no seasonal variation. A high percentage of these sera (62% to 100%) was RESA positive by Modified Indirect Fluorescent Antibody (MIFA), with a significant (p < 0.05) increase of geometric mean titers during the short-rains season, when the transmission of the disease is highest. ELISA with three repetitive RESA peptides (EENV)3 (4x3), (EENVEHDA)2 (8x2) and (DDEHVEEPTVA)2(11x2) did not reveal statistically significant seasonal variations, although a small enhancement of positivity was observed in V. Padaria (15.3 to 38.8%) in the short-rains season with the 8x2 peptides, and with 4x3 and 8x2 peptides in V. Paraíso, with a decrease in 11x2. MIFA titers appeared to be correlated mainly to the peptide 4x3 and it was the immunodominant in the three localities.

Anti-HCV related to HCV PCR and risk factors analysis in a blood donor population of Central Brazil

Data concerning HCV infection in Central Brazil are rare. Upon testing 2,350 voluntary blood donors from this region, we found anti-HCV prevalence rates of 2.2% by a second generation ELISA and 1.4% after confirmation by a line immunoassay. Antibodies against core, NS4, and NS5 antigens of HCV were detected in 81.8%, 72.7%, and 57.5%, respectively, of the positive samples in the line immunoassay. HCV viremia was present in 76.6% of the anti-HCV-positive blood donors. A relation was observed between PCR positivity and serum reactivity in recognizing different HCV antigens in the line immunoassay. The majority of the positive donors had history of previous parenteral exposure. While the combination of ALT>50 IU/l and anti-HBc positivity do not appear to be good surrogate markers for HCV infection, the use of both ALT anti-HCV tests is indicated in the screening of Brazilian blood donors.

Development of a system to monitor surfer’s balance and plantar pressure and surfboard’s movement

O uso da tecnologia tem crescido nas últimas décadas nas mais diversas áreas, seja na indústria ou no dia-a-dia, e é cada vez mais evidente os benefícios que traz. No desporto não é diferente. Cada dia surgem novos desenvolvimentos objetivando a melhoria do desempenho dos praticantes de atividades físicas, possibilitando atingir resultados nunca antes pensados. Além disto, a utilização da tecnologia no desporto permite a obtenção de dados biomecânicos que podem ser utilizados tanto no treinamento quando na melhoria da qualidade de vida dos atletas auxiliando na prevenção de lesões, por exemplo. Deste modo, o presente projeto se aplica na área do desporto, nomeadamente, na modalidade do surfe, onde a ausência de trabalhos científicos ainda é elevada, aliando a tecnologia eletrônica ao desporto para quantificar informações até então desconhecidas. Três fatores básicos de desempenho foram levantados, sendo eles: equilíbrio, posicionamento dos pés e movimentação da prancha de surfe. Estes fatores levaram ao desenvolvimento de um sistema capaz de medi-los dinamicamente através da medição das forças plantares e da rotação da prancha de surfe. Além da medição dos fatores, o sistema é capaz de armazenar os dados adquiridos localmente através de um cartão de memória, para posterior análise; e também enviá-los através de uma comunicação sem fio, permitindo a visualização do centro de pressões plantares; dos ângulos de rotação da prancha de surfe; e da ativação dos sensores; em tempo real. O dispositivo consiste em um sistema eletrônico embarcado composto por um microcontrolador ATMEGA1280; um circuito de aquisição e condicionamento de sinal analógico; uma central inercial; um módulo de comunicação sem fio RN131; e um conjunto de sensores de força Flexiforce. O firmware embarcado foi desenvolvido em linguagem C. O software Matlab foi utilizado para receção de dados e visualização em tempo real. Os testes realizados demostraram que o funcionamento do sistema atende aos requisitos propostos, fornecendo informação acerca do equilíbrio, através do centro de pressões; do posicionamento dos pés, através da distribuição das pressões plantares; e do movimento da prancha nos eixos pitch e roll, através da central inercial. O erro médio de medição de força verificado foi de -0.0012 ± 0.0064 N, enquanto a mínima distância alcançada na transmissão sem fios foi de 100 m. A potência medida do sistema foi de 330 mW.

Studies on human anti-rabies immunization in Brazil: I - Evaluation of the 3 + 1 pre-exposure vaccination schedule under field conditions

The currently used pre-exposure anti-rabies immunization schedule in Brazil is the one called 3+1, employing suckling mouse brain vaccine (3 doses on alternate days and the last one on day 30). Although satisfactory results were obtained in well controlled experimental groups using this immunization schedule, in our routine practice, VNA levels lower than 0.5 IU/ml are frequently found. We studied the pre-exposure 3+1 schedule under field conditions in different cities on the State of São Paulo, Brazil, under variable and sometimes adverse circumstances, such as the use of different batches of vaccine with different titers, delivered, stored and administered under local conditions. Fifty out of 256 serum samples (19.5%) showed VNA titers lower than 0.5 IU/ml, but they were not distributed homogeneously among the localities studied. While in some cities the results were completely satisfactory, in others almost 40% did not attain the minimum VNA titer required. The results presented here, considered separately, question our currently used procedures for human pre-exposure anti-rabies immunization. The reasons determining this situation are discussed.

Studies on human anti-rabies immunization in Brazil: II - Preliminary evaluation of the 2-1-1 schedule for human pre-exposure anti-rabies immunization, employing suckling mouse brain vaccine

This study reports preliminary results of virus neutralizing antibody (VNA) titers obtained on different days in the course of human anti-rabies immunization with the 2-1-1 schedule (one dose is given in the right arm and one dose in the left arm at day 0, and one dose is apllied on days 7 and 21), recommended by WHO for post-exposure treatment with cell culture vaccines. A variant schedule (double dose on day zero and another on day 14) was also tested, both employing suckling mouse brain vaccine. A complete seroconversion rate was obtained after only 3 vaccine doses, and almost all patients (11 of 12) presented titers higher than 1.0 IU/ml. Both neutralizing response and seroconversion rates were lower in the group receiving only 3 doses, regardless of the sample collecting day. Although our results are lower than those found with cell culture vaccines, the geometry mean of VNA is fully satisfactory, overcoming the lower limit recommended by WHO of 0.5 IU/ml. The 2-1-1 schedule could be an alternative one for pre exposure immunization, shorter than the classical 3+1 regimen (one dose on days 0, 2, 4 and 30) with only three visits to the doctor, instead of four.

Application of anti-leptospira ELISA-lgM for the etiologic elucidation of meningitis

Leptospirosis is one of the causes of meningitis, although its importance is not well known. In the present study we contributed to this knowledge by demonstrating specific IgM class anti-leptospira antibodies by the immunoenzymatic method ELISA in 14.6% of cerebrospinal fluid (CSF) samples from 171 patients with meningitis considered to be of indeterminate etiology. The frequencies of positivity were similar in cases with predominance of polymorphonuclear or lymphomononuclear leucocytes in the CSF. Age distribution showed a predominance of the 5 to 15 year age range (72%), and sex distribution showed a predominance of males (68%). The authors discuss the contribution of this method to the etiologic elucidation of meningitis.