Association analysis of human leukocyte antigen class II (DRB1) alleles with leprosy in individuals from São Luís, state of Maranhão, Brazil

Epidemiological studies have demonstrated that the variability of the clinical response to infection caused by Mycobacterium leprae is associated with host genetic factors. The present study investigated the frequency of human leukocyte antigen (HLA) class II (DRB1) alleles in patients with leprosy from São Luís, Maranhão, Brazil. A case-control study was performed in 85 individuals with leprosy and 85 healthy subjects. All samples were analysed via polymerase chain reaction-sequence specific oligonucleotide probes. The HLA-DRB1*16 allele showed a higher frequency in the group with leprosy [(9.41% vs. 4.12%) odds ratio (OR) = 2.41 95% confidence interval (CI) (0.96-6.08) p = 0.05], whereas the HLA-DRB1*11 allele was less frequent in the group with leprosy [(6.47% vs. 11.76%) OR = 0.51 95% CI (0.23-1.12) p = 0.09]. The frequency of HLA-DRB1* alleles between the control group and leprosy patient subgroups presenting different forms of the disease showed that the HLA-DRB1*16 (16.13% vs. 8.24%, OR = 4.10, CI = 1.27-13.27, p = 0.010) and HLA-DRB1*14 (5% vs. 3.53%, OR = 4.63, CI = 1.00-21.08, p = 0.032) alleles were significantly more frequent in patients with different clinical subtypes of leprosy. The sample size was a limitation in this study. Nevertheless, the results demonstrated the existence of a genetic susceptibility associated with the clinical forms of leprosy. The low frequency of the HLA-DRB1*11 allele should be further studied to investigate the possible protective effect of this allele.

Specificity of the rapid rK39 antigen-based immunochromatographic test Kalazar Detect(r) in patients with cutaneous leishmaniasis in Brazil

The aim of this study was to evaluate the specificity of a rapid immunochromatographic test that was developed to detect antibodies against the rK39 antigen for the diagnosis of visceral leishmaniasis (VL). This evaluation was performed using sera from patients with a confirmed diagnosis of active cutaneous leishmaniasis. The sera from 272 patients with a confirmed diagnosis of localised cutaneous leishmaniasis (CL) who resided in an area endemic for Leishmania braziliensis in Brazil were obtained before the initiation of antileishmanial treatment. Kalazar Detect(r)(InBios, Seattle, WA) recombinant K39 antigen-based immunochromatographic strips were used according to the manufacturer's instructions. The test results were evaluated independently by two examiners in sequential order. The positive controls for the test included five serum samples from five patients with parasitologically confirmed diagnosis of VL caused by Leishmania infantum in Brazil. Overall, 100% of the samples obtained from patients with CL were negative, confirming the absence of a serological cross-reaction for individuals with cutaneous disease when these patients were evaluated using the rapid test. The lack of a cross-reaction in patients who were infected by parasites of the same genus highlights the specificity of the rK39 antigen for the diagnosis of VL in areas with the sympatric circulation of L. braziliensis and L. infantum.

Occult hepatitis B virus infection among injecting drug users in the Central-West Region of Brazil

The prevalence of occult hepatitis B virus (HBV) infection was investigated in 149 hepatitis B surface antigen (HBsAg) negative injecting drug users (IDUs) in the Central-West Region of Brazil. Of these individuals, 19 were positive for HBV DNA, resulting in an occult HBV infection prevalence of 12.7% (19/149); six of these 19 individuals had anti-HBV core and/or anti-HBV surface antibodies and 13 were negative for HBV markers. All IDUs with occult hepatitis B reported sexual and/or parenteral risk behaviours. All HBV DNA-positive samples were successfully genotyped. Genotype D was the most common (17/19), followed by genotype A (2/19). These findings reveal a high prevalence of occult HBV infection and the predominance of genotype D among IDUs in Brazil's Central-West Region.

Induction of protective T-helper 1 immune responses against Echinococcus granulosus in mice by a multi-T-cell epitope antigen based on five proteins

In this study, we designed an experiment to predict a potential immunodominant T-cell epitope and evaluate the protectivity of this antigen in immunised mice. The T-cell epitopes of the candidate proteins (EgGST, EgA31, Eg95, EgTrp and P14-3-3) were detected using available web-based databases. The synthesised DNA was subcloned into the pET41a+ vector and expressed in Escherichia coli as a fusion to glutathione-S-transferase protein (GST). The resulting chimeric protein was then purified by affinity chromatography. Twenty female C57BL/6 mice were immunised with the antigen emulsified in Freund's adjuvant. Mouse splenocytes were then cultured in Dulbecco's Modified Eagle's Medium in the presence of the antigen. The production of interferon-γ was significantly higher in the immunised mice than in the control mice (> 1,300 pg/mL), but interleukin (IL)-10 and IL-4 production was not statistically different between the two groups. In a challenge study in which mice were infected with 500 live protoscolices, a high protectivity level (99.6%) was demonstrated in immunised BALB/C mice compared to the findings in the control groups [GST and adjuvant (Adj) ]. These results demonstrate the successful application of the predicted T-cell epitope in designing a vaccine against Echinococcus granulosus in a mouse model.

The use of halloysite clay and carboxyl-functionalised multi-walled carbon nanotubes for recombinant LipL32 antigen delivery enhanced the IgG response

We studied the feasibility of using halloysite clay nanotubes (HNTs) and carboxyl-functionalised multi-walled carbon nanotubes (COOH-MWCNTs) as antigen carriers to improve immune responses against a recombinant LipL32 protein (rLipL32). Immunisation using the HNTs or COOH-MWCNTs significantly increased the rLipL32-specific IgG antibody titres (p < 0.05) of Golden Syrian hamsters. None of the vaccines tested conferred protection against a challenge using a virulent Leptospira interrogans strain. These results demonstrated that nanotubes can be used as antigen carriers for delivery in hosts and the induction of a humoral immune response against purified leptospiral antigens used in subunit vaccine preparations.

Metadados para a descrição de recursos de informação eletrônica: utilização do padrão Dublin Core

Este artigo aborda a necessidade de adoção de padrões de descrição de recursos de informação eletrônica, particularmente, no âmbito da Embrapa Informática Agropecuária. O Rural Mídia foi desenvolvido utilizando o modelo Dublin Core (DC) para descrição de seu acervo, acrescido de pequenas adaptações introduzidas diante da necessidade de adequar-se a especificidades meramente institucionais. Este modelo de metadados baseado no Dublin Core, adaptado para o Banco de Imagem, possui características que endossam a sua adoção, como a simplicidade na descrição dos recursos, entendimento semântico universal (dos elementos), escopo internacional e extensibilidade (o que permite sua adaptação às necessidades adicionais de descrição).

Transgenic tomato plants expressing the antigen gene PfCP-2.9 of Plasmodium falciparum

The objective of this work was to obtain transgenic tomato plants expressing the PfCP-2.9 protein (a chimera of the antigens MSP1 and AMA1 of Plasmodium falciparum). Cotyledons of seven-day-old tomatoes, cultivar Summers, were transformed via Agrobacterium tumefaciens. Transgenic expression in the T0 plants was verified in the DNA extracted from fruits. PCR analysis was used to test the presence of the gene of interest in the T1 generation. Reverse transcriptase PCR provided evidence of gene expression at the RNA level, and Western blot analysis confirmed the presence of the protein of interest in the T1 plants. This is the first report of successful transformation with the expression of a malaria antigen (PfCP-2.9) in transgenic tomato plants from the T0 and T1 generations.

Comparação entre fragmentos obtidos com agulhas de calibres 14 e 12 em "core biopsy" estereotáxica de lesões mamárias impalpáveis: diferenças entre o tamanho dos fragmentos e freqüência dos tipos de lesões diagnosticadas

Com o objetivo de avaliar o desempenho das agulhas de calibres 14 e 12 na obtenção de "core biopsies" da mama sob controle estereotáxico, os autores compararam biópsias de lesões mamárias impalpáveis obtidas com agulhas desses calibres quanto ao tamanho dos fragmentos e freqüência dos tipos de lesões diagnosticadas. De um total de 1.009 biópsias, os autores analisaram 900 obtidas por agulhas 14 e 109 obtidas por agulhas 12, para análise da freqüência das lesões. Os tamanhos dos fragmentos foram analisados nos 700 primeiros casos para os espécimes obtidos por agulhas 14. A utilização de agulhas 12 resultou em maior tamanho dos fragmentos (p < 0,001), ausência de material insatisfatório, redução na proporção das lesões benignas não-neoplásicas (p = 0,03) e aumento na freqüência de carcinomas (p < 0,001), tanto in situ como invasivos. Tais achados podem refletir aumento da acurácia do diagnóstico, em virtude da maior quantidade de material e/ou melhor seleção radiológica das biópsias obtidas por agulhas 12, pela utilização coincidente da mamografia digital em campo limitado (estereotaxia digital).

Estudo prospectivo de 100 casos de "core" biópsia dirigida por ultra-som e revisão da literatura

OBJETIVO: Mostrar a eficácia do método para evitar biópsias excisionais, verificar suas dificuldades técnicas, definir entre seguimento e biópsia excisional nos nódulos categoria IV do BI-RADS™ e agilizar o procedimento cirúrgico nos casos de nódulos altamente suspeitos de malignidade (categoria V). MATERIAIS E MÉTODOS: As pacientes foram submetidas a exame clínico de rotina, mamografia e ultra-som. A "core" foi feita com pistola automática e agulha número 14, e foram colhidas de quatro a oito amostras. RESULTADOS: Das 100 lesões estudadas, 47 foram submetidas à cirurgia, além da "core", e diagnosticaram-se 34 carcinomas (34,0%). Das 23 lesões classificadas como categoria III, identificou-se apenas um carcinoma (4,34%); das 43 classificadas como categoria IV, sete (16,28%); e das 34 classificadas como categoria V, 26 (76,47%). Evitou-se a biópsia excisional em 53 casos (53,0%). Identificou-se dificuldade no método da "core" em sete casos (7,0%), devido a material insuficiente, risco para malignidade envolvendo lesões esclerosantes complexas e discordância entre imagem e histologia. As 33 lesões com resultados de malignidade à punção biópsia por agulha grossa foram confirmadas após a biópsia cirúrgica. Em um caso o diagnóstico pela "core" foi de hiperplasia ductal atípica e após a biópsia cirúrgica da peça diagnosticou-se carcinoma, correspondendo a um resultado falso-negativo. Não houve nenhum resultado falso-positivo.

Step-by-step of ultrasound-guided core-needle biopsy of the breast: review and technique

Ultrasound-guided core-needle biopsy has high sensitivity in the diagnosis of breast cancer. The present study is aimed at detailing the main steps of such procedure, including indications, advantages, limitations, follow-up and description of the technique, besides presenting a checklist including the critical steps required for an appropriate practice of the technique. In the recent years, an increasing number of patients have required breast biopsy, indicating the necessity of a proportional increase in the number of skilled professionals to carry out the procedures and histological diagnoses. A multidisciplinary approach involving the tripod clinical practice-radiology-pathology is responsible for the highest rate of accuracy of the technique and must always be adopted.

Diagnostic underestimation of atypical ductal hyperplasia and ductal carcinoma in situ at percutaneous core needle and vacuum-assisted biopsies of the breast in a Brazilian reference institution

Abstract Objective: To determine the rates of diagnostic underestimation at stereotactic percutaneous core needle biopsies (CNB) and vacuum-assisted biopsies (VABB) of nonpalpable breast lesions, with histopathological results of atypical ductal hyperplasia (ADH) or ductal carcinoma in situ (DCIS) subsequently submitted to surgical excision. As a secondary objective, the frequency of ADH and DCIS was determined for the cases submitted to biopsy. Materials and Methods: Retrospective review of 40 cases with diagnosis of ADH or DCIS on the basis of biopsies performed between February 2011 and July 2013, subsequently submitted to surgery, whose histopathological reports were available in the internal information system. Biopsy results were compared with those observed at surgery and the underestimation rate was calculated by means of specific mathematical equations. Results: The underestimation rate at CNB was 50% for ADH and 28.57% for DCIS, and at VABB it was 25% for ADH and 14.28% for DCIS. ADH represented 10.25% of all cases undergoing biopsy, whereas DCIS accounted for 23.91%. Conclusion: The diagnostic underestimation rate at CNB is two times the rate at VABB. Certainty that the target has been achieved is not the sole determining factor for a reliable diagnosis. Removal of more than 50% of the target lesion should further reduce the risk of underestimation.

LC-MS/MS method applied to preclinical pharmacokinetic investigation of olanzapine-loaded lipid-core nanocapsules

In spite of different methods reported in the literature to determine olanzapine in biological fluids, all of them used high volumes of plasma. Therefore, the purpose of this paper was to develop an LC-MS/MS method using small plasma volume (0.1 mL) to apply in a preclinical pharmacokinetic investigation. The method was linear over the concentration ranges of 10 - 1000 ng mL-1. Extraction recoveries, stability, and validation parameters were evaluated. Results were within the acceptable limits of international guidelines. A significant decrease in clearance led to a significant 2.26-times increase in AUC0 - 6h of olanzapine-loaded lipid-core nanocapsules compared with free-olanzapine.

CHARACTERIZING THE MECHANISM OF QUETIAPINE DISTRIBUTION IN LIPID-CORE NANOCAPSULES PSEUDO-PHASES USING A VALIDATED LC/UV METHOD

Quetiapine is an atypical antipsychotic used to treat schizophrenia. However, despite great interest for its chronic therapeutic use, quetiapine has some important side effects such as weight gain induction. The development of a quetiapine nanocarrier can potentially target the drug into central nervous system, resulting in a reduction of systemic side effects and improved patient treatment. In the present work, a simple liquid chromatography/ultraviolet detection (LC/UV) analytical method was developed and validated for quantification of total quetiapine content in lipid core nanocapsules as well as for determination of incorporation efficiency. An algorithm proposed by Oliveira et al. (2012) was applied to characterize the distribution of quetiapine in the pseudo-phases of the nanocarrier, leading to a better understanding of the quetiapine nanoparticles produced. The analytical methodology developed was specific, linear in the range of 0.5 to 100 µg mL−1 (r2 > 0,99), and accurate and precise (R.S.D < ±5%). The absolute recovery of quetiapine from the nanoparticles was approximately 98% with an incorporation efficiency of approximately 96%. The results indicated that quetiapine was present in a type III distribution according to the algorithm, and was mainly located in the core of the nanoparticle because of its logD in the formulation pH (6.86 ± 0.4).

O Core Curriculum da Unesco como Base para Formação em Bioética

RESUMO A revolução biotecnológica das últimas décadas teve como resultado o desenvolvimento de um poder quase sem limites sobre a vida humana. Tal contexto exige do profissional uma visão globalizada dos problemas éticos e sociais da era contemporânea, alicerçada em sólidas bases filosóficas e legais. Este contexto torna necessária a promoção de novas competências e habilidades relacionadas à vida profissional. Neste sentido, o ensino da Bioética desponta como uma possibilidade de inovação curricular alternativa ao tradicional modelo prescritivo e normativo. Este artigo relata a experiência da Cátedra Unesco de Bioética da Universidade de Brasília com a utilização do Core Curriculum proposto pela Unesco como instrumento didático-pedagógico adequado ao ensino da Bioética. Entre os dilemas pedagógicos enfrentados pela Bioética como disciplina encontram-se: a construção de seus conteúdos, sua estruturação, as concepções teóricas a serem seguidas e seus objetivos. A contextualização e o aperfeiçoamento da estratégia proposta pelo Core Curriculum podem significar importantes instrumentos facilitadores para docentes que buscam organizar práticas didático-pedagógicas inovadoras em Bioética com o intuito de proporcionar resultados efetivos na formação de seus estudantes.