Abstract Objectives: Upper respiratory tract infections in children generally have significant morbidity and mortality. There is little data available about functional immaturity of the immune system and the child’s susceptibility to infections at the beginning of their lives, thus, justifying a more specific immunological analysis. Method: Analysis of hemograms and innate and adaptive immune responses in 95 children between age 1 to 6 years with episodes of recurrent respiratory infections (test group n = 39) and without these episodes (control group n = 56) was carried out. The production of reactive oxygen intermediates by peripheral blood cells stimulated by phorbol myristate acetate was analyzed. Additionally, the number of B lymphocytes, auxiliary T lymphocytes, and cytotoxic cells was determined using flow cytometry. Results: Results from both groups did not show statistically significant differences in red blood cells, total leukocytes count, and the differential neutrophils, eosinophils, basophils, lymphocytes, and monocytes count. The analysis of the number of B lymphocytes, auxiliary T lymphocytes (LTCD4), and cytotoxic cells (LTCD8) also did not show any difference between both groups. However, the production of radical oxygen intermediates was significantly reduced in the test group as compared to the control group (p < 0.05). Conclusions: There was no difference in the analysis of hemograms, leukograms, or the number of lymphocytes, LTCD4, LTCD8, or LTCD19. The reduced production of oxygen intermediates in the affected group suggests that these children’s microbicide capacity is compromised, which may be related to their recurrent respiratory infections.

Abstract Background: Triatomines are blood-feeding arthropods belonging to the subfamily Triatominae (Hemiptera; Reduviidae), capable of producing immunomodulatory and water-soluble molecules in their hemolymph, such as antimicrobial peptides (AMPs). In this work, we evaluated the antifungal and immunomodulatory activity of the hemolymph of Meccus pallidipennis (MPH) and Rhodnius prolixus (RPH) against Cryptococcus neoformans. Methods: We assessed the activity of the hemolymph of both insects on fungal growth by a minimum inhibitory concentration (MIC) assay. Further, RAW 264.7 macrophages were cultivated with hemolymph and challenged with C. neoformans. Then, their phagocytic and killing activities were assessed. The cytokines MCP-1, IFN-γ, TNF-α, IL-10, IL-12, and IL-6 were measured in culture supernatants 4- and 48-hours post-infection. Results: Both hemolymph samples directly affected the growth rate of the fungus in a dose-dependent manner. Either MPH or RPH was capable of inhibiting fungal growth by at least 70%, using the lowest dilution (1:20). Treatment of RAW 264.7 macrophages with hemolymph of both insects was capable of increasing the production of MCP-I and TNF-α. In addition, when these cells were stimulated with hemolymph in the presence of C. neoformans, a 2- and a 4-fold increase in phagocytic rate was observed with MPH and RPH, respectively, when compared to untreated cells. For the macrophage killing activity, MPH decreased in approximately 30% the number of viable yeasts inside the cells compared to untreated control; however, treatment with RPH could not reduce the total number of viable yeasts. MPH was also capable of increasing MHC-II expression on macrophages. Regarding the cytokine production, MCP-I and TNF-α, were increased in the supernatant of macrophages treated with both hemolymphs, 4 and 48 hours after stimulation. Conclusion: These results suggested that hemolymph of triatomines may represent a source of molecules capable of presenting antifungal and immunomodulatory activity in macrophages during fungal infection.

Abstract Background Mycobacterium leprae and Mycobacterium lepromatosis are gram-positive bacterial pathogens and the causative agents of leprosy in humans across the world. The elimination of leprosy cannot be achieved by multidrug therapy alone, and highlights the need for new tools and drugs to prevent the emergence of new resistant strains. Methods In this study, our contribution includes the prediction of vaccine targets and new putative drugs against leprosy, using reverse vaccinology and subtractive genomics. Six strains of Mycobacterium leprae and Mycobacterium lepromatosis (4 and 2 strains, respectively) were used for comparison taking Mycobacterium leprae strain TN as the reference genome. Briefly, we used a combined reverse vaccinology and subtractive genomics approach. Results As a result, we identified 12 common putative antigenic proteins as vaccine targets and three common drug targets against Mycobacterium leprae and Mycobacterium lepromatosis. Furthermore, the docking analysis using 28 natural compounds with three drug targets was done. Conclusions The bis-naphthoquinone compound Diospyrin (CID 308140) obtained from indigenous plant Diospyros spp. showed the most favored binding affinity against predicted drug targets, which can be a candidate therapeutic target in the future against leprosy.

Ilustramos aqui um caso de uma apresentação atípica (na forma de linfadenomegalia e febre) de uma das doenças zoonóticas mais comuns no mundo - brucelose - em um paciente brasileiro de 22 anos (chefe de cozinha) que retornara ao Brasil recentemente após ter morado e viajado na Europa por um ano. A histopatologia, a história clínica e a resposta ao tratamento foram consistentes com o diagnóstico de brucelose, que foi confirmada por PCR em uma amostra de urina. Também revisamos alguns aspectos da brucelose, como manifestações clínicas, diagnóstico e tratamento.

This case illustrates a rare presentation (as lymphadenopathy and fever) of one of the most common zoonotic diseases worldwide-brucellosis-in a 22-year-old Brazilian male (a chef) who had recently returned to Brazil after having lived in and traveled around Europe for one year. The histopathology, clinical history, and response to treatment were all consistent with a diagnosis of brucellosis, which was confirmed by PCR in a urine sample. We also review some aspects of brucellosis, such as the clinical features, diagnosis, and management.

O objetivo do estudo consistiu em detectar seqüências no ADNr e as suas regiões no Histoplasma capsulatum, que pudessem ser consideradas espécie-específicas e usadas como método molecular para o diagnóstico pela técnica da reação em cadeia da polimerase aninhada ("nested PCR") com seqüências específicas ("primers") para H. capsulatum: regiões 18S ADNr (HC18), 100kDa (HC100) e a seqüência 5.8 S ADNr-ITS (HC5.8). A "nested PCR" com as seqüências HC18, HC100 e HC5.8 resultaram em 100% de especificidade. Os ensaios moleculares podem aumentar a especificidade, sensibilidade e rapidez na diagnose da Histoplasmose.

The aim of the study was to detect the rDNA sequences and their regions in Histoplasma capsulatum, which could be considered species-specific and used as a molecular method for this diagnosis by the technique of nested polymerase chain reaction (nested PCR), employing specific sequences (primers) for H. capsulatum: 18S rDNA region (HC18), 100 kDa (HC100) and the sequence 5.8 S-ITS rDNA (HC5.8). The PCR sequences HC18, HC100 and HC5.8 resulted in a specificity of 100%. The molecular assays may increase the specificity, sensitivity and speed in the diagnosis of Histoplasmosis.

A tungíase é doença parasitária da pele, comum no Brasil, causada pela pulga Tunga penetrans. Realizamos estudo transversal para avaliar a ocorrência da infestação por T. penetrans em crianças, da pré-escola e do ensino fundamental de primeira à quarta série da rede municipal de ensino de Criciúma, Santa Catarina, Sul do Brasil. No total, 917 crianças (6 - 10 anos) de 68 escolas foram incluídas no estudo (11% da população alvo). As crianças foram submetidas à inspeção física e nas com tungíase foi realizada visita domiciliar, para avaliar as condições da casa e para entrevista com o responsável legal da criança. Das 917, 15 (1,6%) crianças estavam parasitadas. A prevalência do parasitismo foi duas vezes maior no sexo feminino (10 crianças - 2,2%) que no masculino (cinco crianças - 1,1%), mas pelo pequeno número de parasitados a diferença foi estatisticamente não significante (p = 0,18). A média aritmética das lesões encontradas foi de 1,4 por criança. Em uma criança foi observada infecção secundária bacteriana. Das 15 crianças parasitadas, 10 (66,7%) estudavam em escolas construídas em solo piritoso. As famílias de 12 crianças parasitadas (79,9%) residiam no local há menos de oito anos e foram consideradas famílias desfavorecidas, tendo o chefe da família baixa escolaridade. Concluímos que, em Criciúma, a tungíase ocorre em crianças da pré-escola e do ensino fundamental de primeira à quarta série da rede municipal de ensino, com baixa prevalência e carga parasitária.

Tungiasis is a common parasitic skin disease in Brazil, caused by the flea Tunga penetrans. We conducted a cross-sectional study to evaluate the occurrence of infestation with T. penetrans in primary schoolchildren in Criciúma, Santa Catarina State (South Brazil). In total, 917 children (6-10 years) of 68 schools were included in the study (11% of target population). Physical examination was followed by residential visits of those infested, to assess housing conditions and to interview the children's guardians. Of the 917, 15 (1.6%) children were infested with the parasite. The prevalence was twice as high in females (10 children - 2.2%) as compared to males (five children - 1.1%), but due to the low number of infested individuals, the difference was statistically not significant (p = 0.18). The mean number of lesions found was 1.4 per child. One child presented bacterial superinfection. Of the 15 children infested, 10 (66.7%) studied in schools built on pyritiferous soil. The families of 12 children (79.9%) had been living in the area in which the study was conducted for less than eight years and were considered underprivileged families with the head of the household having a low education status. We conclude that in Criciúma tungiasis occurs in primary school children, but prevalence and parasite load are low.

O objetivo deste estudo foi verificar a freqüência da infecção pelo HIV em profissionais do sexo, atuantes em Imbituba (SC), identificar o subtipo viral e a suscetibilidade do vírus aos medicamentos antiretrovirais. De dezembro de 2003 a fevereiro de 2004, foram entrevistadas 90 mulheres, profissionais do sexo, e a freqüência de HIV nessa população foi de 6,7%. O teste de genotipagem para o HIV, realizado em quatro amostras, detectou em três delas o subtipo C, que é predominante na África e Ásia, e em uma o subtipo B, prevalente no Brasil, EUA e Europa. Os resultados sugerem que o Porto de Imbituba pode ser uma das portas de entrada para do HIV-1 subtipo C no Brasil, e a partir dessa localidade ocorrer sua disseminação para o restante do País e países do Mercosul pela rodovia BR-101. Isto aponta para a necessidade de trabalhos de prevenção, com a finalidade de reduzir a introdução, transmissão e disseminação do HIV subtipo C.

The objective of this study was to investigate the frequency of HIV infection among female sex workers in the port area of Imbituba (State of Santa Catarina), and to identify the viral subtype and its susceptibility to antiretroviral medications. Ninety women were interviewed between December 2003 and February 2004. Six (6.7%) were HIV-positive. Genotyping for HIV, performed on four samples, detected subtype C in three of them, which is predominant in Africa and Asia, and subtype B in one of them, which is prevalent in Brazil, USA and Europe. The results suggest that the Port of Imbituba may be one of the gateways for HIV-1 subtype C to enter Brazil, and for its dissemination to the rest of the country and the Mercosul area, along the highway BR-101. This points towards the need for preventive work to reduce the introduction and dissemination of HIV subtype C in Brazil.

We examined HIV frequency and probable risk factors among female sex workers in the port city of Imbituba, Southern Brazil. From December 2003 through February 2004, 90 female sex workers were interviewed in order to investigate demographic, socioeconomic and behavioral variables related to HIV infection. A blood sample of each woman was also collected to test for HIV antibodies. Six (6.7%) of the sex workers were HIV-positive; the significantly-correlated risk factors were the daily number of clients (p = 0.008), the use of inhaled illicit drugs by the sex workers (P = 0.053) and by their clients (p = 0.005), and the lack of condom use in sexual relations (p = 0.015). The HIV infection rate in these sex workers was higher than that in the general population and similar to that found in other studies made with Brazilian populations presenting the same characteristics. This highlights the need for preventative measures, especially in this port area, in order to reduce transmission and to deter the introduction and dissemination of HIV.

Síndrome pulmonar por hantavírus. Relato dos três primeiros casos em São Paulo, Brasil O síndrome pulmonar por hantavírus foi reconhecido a partir dos casos ocorridos nos EUA em 1993. Esse alerta serviu tanto para os médicos americanos como para os de outros países na identificação de casos dessa doença. Na cidade de São Paulo, Brasil, em 1993, foram registrados 3 casos com essa manifestação da hantavirose; eram irmãos, do sexo masculino, jovens, residentes na região da Mata Atlântica, com desmatamento recente. Dois dos casos evoluíram para a morte causada por insuficiência respiratória aguda e o terceiro com recuperação sem seqüelas. No paciente sobrevivente o diagnóstico foi estabelecido por critério laboratorial pela pesquisa de anticorpos específicos das classes IgM e IgG pela técnica de imunofluorescência indireta. Dos casos que evoluíram para o óbito, um foi confirmado laboratorialmente pela técnica de imuno-histoquímica para hantavírus em tecido, em cortes histológicos de pulmão e coração; e no outro o diagnóstico foi clínico e epidemiológico

The hantavirus pulmonary syndrome was first recognized in cases that occurred in the U.S. in 1993, which served as an alert not only for American physicians but also for physicians in other countries for the identification of the disease. In the city of São Paulo, Brazil, 3 cases of the syndrome were recorded in 1993. The patients were young brothers residing in the Mata Atlântica (Atlantic Forest) region submitted to recent deforestation. Two of the patients died of acute respiratory insufficiency and the third recovered without sequelae. In the surviving patient the diagnosis was established by a laboratory criterion based on the detection of specific IgM and IgG class antibodies by indirect immunofluorescence. In the two patients who died, the diagnosis was confirmed by laboratory tests using immunoperoxidase technique for hantavirus in tissue, in histological lung and heart sections in one case, and by clinical and epidemiological data in the other

A detecção de IgE apresenta dificuldades técnicas pela reduzida concentração que se encontra no LCR e no soro. Utilizando método imunoenzimático com conjugado anti-IgE-fosfatase alcalina e substrato fluorigênico, foram estudadas 86 amostras de LCR de três grupos: A) 29 pacientes com NC, B) 36 pacientes com afecções neurológicas diversas (neurossífilis, neurotuberculose, meningites, tumores, hemorragias) e C) 21 indivíduos de pós-alta médica de meningites bacterianas. Os resultados obtidos foram: A) 0,05 a 3,00 Ul/ml (0,76±0,79), B) 0,00 a 1,50 Ul/ml (0,23±0,34) e C) 0,05 a 1,25 Ul/ml (0,34±0,34). Os resultados obtidos sugerem que a IgE parece ter papel na patogenia da NC e esforços devem ser feitos para padronização de teste para detecção de anticorpos IgE específicos.

The detection of IgE is technically difficult because of its reduced concentrations in serum, and even lower concentrations in cerebrospinal fluid (CSF). In the present investigation we studied 86 CSF samples using an immunoenzymatic method with an anti-IgE-alkaline phosphatase conjugate and a fluorigenic substrate. The samples were from three groups: A) 29 patients with neurocysticercosis (NC), B) 36 patients with different neurologic disorders (neurosyphilis, neurotuberculosis, meningitis, tumors, hemorrhage) and C) 21 discharged individuals who had been hospitalized for bacterial meningitis. The results obtained were: A) 0.05 to 3.00 IU/ml (0.76±0.79), B) 0.00 to 1.50 IU/ml (0.23±0.34) and C) 0.05 to 1.25 IU/ml (0.34±0.34). The present results suggest that IgE appears to play a role in the pathogeny of NC and that efforts should be made to standardize a test for the detection of specific IgE antibodies.

Com a finalidade de melhorar o diagnóstico da leptospirose humana, padronizou-se o teste dot-ELISA para a pesquisa de anticorpos específicos da classe IgM na saliva. Empregaram-se amostras de saliva e soro coletadas simultaneamente de 20 pacientes com a forma ictero-hemorrágica da doença, de 10 pacientes com outras patologias e 5 controles negativos. Culturas de Leptos-pira em meio EMJH, dos sorovares: icterohaemorrhagiae, canicola, hebdomadis, brasiliensis e cynopteri, foram utilizadas como antígeno, na concentração proteica individual de 0,2 µg/µl, misturadas em volumes iguais. Na fase sólida do teste empregou-se tecido de po-liéster impregnado com resina de N-metilol-acrilamida. O volume do antígeno para cada teste foi de 1µl, o de saliva 8µl, o de conjugado anti-IgM humana marcada com peroxidase, de 30µl. A leitura foi visual, após revelação em solução cromógena de preparo recente. O suporte de tecido é de fácil obtenção e manuseio, diferente do clássico de membrana de nitrocelulose. A saliva pode ser coletada diretamente sobre o suporte, o que facilita o método e diminui gastos e riscos inerentes à coleta e processamento do sangue.

In order to improve the diagnosis of human leptospirosis, we standardized the dot-ELISA for the search of specific IgM antibodies in saliva. Saliva and serum samples were collected simultaneously from 20 patients with the icterohemorrhagic form of the disease, from 10 patients with other pathologies and from 5 negative controls. Leptospires of serovars icterohaemorrhagiae, canicola, hebdomadis, brasiliensis and cynopteri grown in EMJH medium and mixed together in equal volumes, were used as antigen at individual protein concentration of 0.2 µg/µl. In the solid phase of the test we used polyester fabric impregnated with N-methylolacrylamide resin. The antigen volume for each test was 1µl, the saliva volume was 8 µl, and the volume of peroxidase-labelled anti-human IgM conjugate was 30 µl. A visual reading was taken after development in freshly prepared chromogen solution. In contrast to the classic nitrocellulose membrane support, the fabric support is easy to obtain and to handle. Saliva can be collected directly onto the support, a fact that facilitates the method and reduces the expenses and risks related to blood processing.

Realizou-se estudo comparativo de diferentes sorotipos de Leptospira interrogans utilizados na preparação de antígenos empregados no teste ELISA, para a detecção de anticorpos da classe IgM, em amostras de soro na fase precoce e tardia da leptospirose humana. Foram utilizados dez sorotipos, escolhidos entre os que apresentaram maior reatividade na soroaglutinação microscópica (SAM), na cidade de São Paulo. Os cinco sorotipos que apresentaram melhores resultados individualmente no teste ELISA-IgM (canicola, hebdomadis, icterohaemorrhagiae, cynopteri e brasiliensis), foram também estudados em mistura antigênica. Os antígenos não tratados apresentaram maior reatividade do que os antígenos tratados com Triton X - 100 (4%) à temperatura de 50ºC, durante 4 horas. O teste ELISA-IgM empregando os sorotipos não tratados, isoladamente, e em mistura antigênica, mostrou-se altamente sensível, podendo ser empregado como teste de triagem para o diagnóstico precoce da leptospirose humana. Outra aplicação do teste é permitir a detecção do início de situações epidêmicas ou de surtos, possibilitando acionar medidas de vigilância epidemiológica.

A comparative study among different serovars of Leptospira interrogans was performed in order to prepare antigens to detect IgM antibodies by ELISA in early and late phase of human leptospirosis. Ten serovars were chosen among the most prevalent detected by microscopic seroagglutination (SAM) in São Paulo city. Using ELISA-IgM five of them showed better results (canicola, hebdomadis, icterohaemorrhagiae, cynopteri and brasiliensis). These ones were also studied in a pool. The non-treated antigens showed higher reactivity than the Triton X-100 (4%/50ºC/4h). ELISA-IgM using individually or pool of non treated antigens proved to be reliable with high sensitivity and should be used for an earlier diagnosis of leptospirosis, as a trial test. Faster diagnostic elucidation can be useful to detect epidemic situations, so, allowing epidemiological surveillance interventions.

Foi padronizada e avaliada a reação de hemaglutinação passiva (RHA) para pesquisa de anticorpos específicos, anti-Cysticercus cellulosae, no líquido cefalorraquiano (LCR). Foram utilizadas hemácias humanas O Rh-formolizadas e sensibilizadas com extrato antigênico salino total de cisticercos, ainda pouco estudado. De 115 amostras estudadas de LCR de pacientes com neurocisticercose, 94 foram reagentes, resultando em 81,7% de sensibilidade, com intervalo de confiança de 95% de probabilidade (IC95%) abrangendo de 74,5% e 88,9%. Também foram ensaiadas 89 amostras de LCR de indivíduos do grupo controle, sendo tão reagentes em 94,4%, com IC95%, de 89,6% a 99,2%. Os valores preditivos positivo e negativo obtidos para a RHA foram, respectivamente, de 1,4% e 99,9%, considerando a prevalência média de neurocisticercose na América Latina de 0,1%. Os resultados indicam que a RHA como um método simples, altamente reprodutível e moderadamente sensível para a detecção de anticorpos específicos no LCR, porém apropriados para a triagem de infectados.

A passive haemagglutination test (PHA) for human neurocysticercosis was standardized and evaluated for the detection of specific antibodies to Cysticercus cellulosae in cerebrospinal fluid (CSF). For the assay, formaldehyde-treated group O Rh-human red cells coated with the cysticerci crude total saline extract (TS) antigen were employed. A total of 115 CSF samples from patients with neurocysticercosis was analysed, of these 94 presented reactivity, corresponding to 81.7% sensitivity, in which confidence limit of 95% probability (CL95%) ranged from 74.5% to 88.9%. Eighty-nine CSF samples derived from individuals of control group presented as nonreactive in 94.4% (CL95% from 89.6% to 99.2%). The positive and negative predictive values were 1.4% and 99.9%, respectively, considering the mean rate of that this assay provide a rapid, highly reproducible, and moderately sensitive mean of detecting specific antibodies in CSF samples.