Refinement of the rKLi8.3-Based Serodiagnostic ELISA Allows Detection of Canine Leishmaniosis in Dogs with Low Antibody Titers.

The diagnosis of canine leishmaniasis (CanL) still represents a challenge due to the variable clinical manifestations and the large number of asymptomatic dogs. Serological tests are most commonly used to detect infected animals, revealing anti-Leishmania antibodies, mainly of the IgG isotype. Recently, a new diagnostic antigen, rKLi8.3, containing 8.3 kinesin tandem repeats (TR) from a Leishmania infantum strain from Sudan, has been shown to provide excellent specificity and sensitivity for the detection of Leishmania-infected humans and dogs. However, asymptomatic animals with very low antibody titers are often difficult to detect by serodiagnosis. Thus, we wondered whether the addition of an anti-IgG-enhancing step in the protein A/G-based rKLi8.3-ELISA will improve the diagnostic performance without decreasing the specificity. For this, parasitologically confirmed CanL cases with low or high clinical scores, uninfected healthy controls and dogs with other infections were tested by rKLi8.3-ELISA as well as two different immunochromatographic rapid tests, rKLi8.3-lateral flow test (LFT) and Dual Path Platform (DPP®) based on the rK28 antigen. Our results show that the diagnostic accuracies of the rKLi8.3-ELISA and LFT were similar to that of DPP, missing several asymptomatic animals. However, the addition of a secondary, amplifying anti-dog IgG antibody in the protein A/G-based rKLi8.3-ELISA enabled the detection of nearly all asymptomatic dogs without compromising its specificity.

Comparative Study of a Novel Lateral Flow Rapid Test with Conventional Serological Test Systems for the Diagnosis of Canine Leishmaniosis in Croatia and Brazil.

Control of canine infections with Leishmania infantum (L. infantum), a major zoonotic disease in Brazil and southern Europe, is becoming increasingly important due to its close proximity to humans, the increasing import of dogs from endemic regions and the impact of climate change on vector spreading. Simple, rapid and reliable diagnostic tests are therefore needed to detect infected dogs. Here, we re-evaluated different serological methods for the diagnosis of canine leishmaniosis (CanL) in Croatia and Brazil. The diagnostic performance of the indirect fluorescent antibody test (IFAT) and the VetLine® Leishmania ELISA (GSD Frankfurt, Germany) was compared with three rKLi8.3-based diagnostic test systems, the rKLi8.3 ELISA (GSD Frankfurt, Germany), the INgezim® Leishma CROM (GSD Madrid, Spain) lateral flow test (LFT) and the VetBlot®Leishmania LineBlot (GSD Frankfurt, Germany). CanL symptomatic dogs were efficiently diagnosed by all tests, except the VetLine® Leishmania ELISA, which is based on whole Leishmania antigens. The advantage of rKLi8.3 was also observed in oligo- and asymptomatic dogs from Brazil and Croatia, although with reduced diagnostic efficiency compared to symptomatic dogs. Similar to IFAT and rKLi8.3 ELISA, the LFT did not cross-react with other common canine pathogens; it showed very high specificity for healthy dogs from endemic regions in both countries and did not react with healthy, vaccinated dogs in Brazil. In conclusion, serodiagnostic tests based on the rKLi8.3 antigens are superior to whole parasite antigens, and the LFT has the advantage of providing a laboratory-independent, rapid and specific diagnosis of CanL.

Efficiency of rLb6H recombinant protein from Leishma-nia (Viannia) braziliensis for the detection of canine visceral leishmaniasis / Eficiência da proteína recombinante rLb6H de Leishmania (Viannia) brazi-liensis na detecção da leishmaniose visceral canina

Introduction: Visceral leishmaniasis (VL) is a serious endemic disease in many tropical and subtropical countries, with a strong incidence in Brazil. The disease is transmitted by the bite of infected female phlebotomine sandflies, with dogs being the main urban reservoirs of the parasite. The diverse clinical profile and the long incubation period are challenges for the diagnosis of canine visceral leishmaniasis (CVL). Recombinant proteins from Leishmania spp. have been studied as antigens that can increase the accuracy of serological tests. Objective: To evaluate the diagnostic performance of the recombinant protein rLb6H, from Leishmania braziliensis, in comparison to the reference antigens rK39 and rK28, from L. donovani, prioritizing the identification of subclinical infected dogs. Material and Methods: Serum IgG reactivity to rLb6H, rK28, and rK39 recombinant proteins was assessed in dogs with previously parasitological confirmation of CVL, subdivided according to their clinical status, using immunoenzymatic assay (ELISA). Diagnostic accuracy of each ELISA was evaluated by receiver operating characteristic (ROC) curve analysis. Results: While all antigens showed a better performance in detecting CVL in symptomatic dogs (SD), detection of CVL in the oligosymptomatic (OD) and asymptomatic (AD) groups was lower, but rLb6H achieved high sensitivity for asymptomatic CVL. Interestingly, the most reactive CVL samples to rK28 were barely detected by rLb6H, while the less reactive to rK28, mostly from the AD group, presented higher reactivity to rLb6H. Conclusion: The recombinant protein rLb6H showed utility in the detection of asymptomatic CVL, displaying a complementary reactivity to rK39 and rK28. Thus, these results suggest that rLb6H could be incorporated into multi-antigen strategies, to increase diagnostic accuracy of CVL.

Introdução: A leishmaniose visceral (LV) é uma doença endêmica grave em muitos países tropicais e subtropicais, tendo forte incidência no Brasil. A doença é transmitida pela picada de flebotomíneos fêmeas infectadas, sendo os cães os principais reservatórios urbanos do parasito. O perfil clínico diversificado e o longo período de incubação são desafios para o diagnóstico da leishmaniose visceral canina (LVC). Proteínas recombinantes de Leishmania spp. têm sido estudadas como antígenos que podem aumentar a precisão de testes sorológicos. Objetivo: Avaliar o desempenho diagnóstico da proteína recombinante rLb6H, de Leishmania braziliensis, em comparação com os antígenos de referência rK39 e rK28, de L. donovani, priorizando a identificação de cães com infecção subclínica. Material e Métodos: A reatividade de anticorpos IgG séricos às proteínas recombinantes rLb6H, rK28 e rK39 foi avaliada em cães com confirmação parasitológica prévia de LVC, subdivididos de acordo com seu quadro clínico, utilizando ensaio imunoenzimático (ELISA). A precisão diagnóstica de cada ELISA foi avaliada pela análise da curva ROC (receiver operating characteristic curve). Resultados: Enquanto todos os antígenos mostraram um melhor desempenho na detecção de CVL em cães sintomáticos (SD), a detecção de CVL nos grupos oligossintomáticos (OD) e assintomáticos (AD) foi menor, mas rLb6H alcançou alta sensibilidade para CVL assintomática. Curiosamente, as amostras de CVL mais reativas a rK28 foram pouco detectadas por rLb6H, enquanto as menos reativas a rK28, principalmente do grupo AD, apresentaram maior reatividade a rLb6H. Conclusão: A proteína recombinante rLb6H mostrou utilidade na detecção de CVL assintomática, apresentando uma reatividade complementar a rK39 e rK28. Assim, estes resultados sugerem que o rLb6H pode ser incorporado em estratégias multi-antígeno para aumentar a acurácia diagnóstica da leishmaniose visceral.

Serum IgA Antibodies Specific to M. leprae Antigens as Biomarkers for Leprosy Detection and Household Contact Tracking.

Leprosy remains endemic in several developing countries, such as India and Brazil, in part due to delayed diagnosis that facilitates ongoing transmission. Although immunoglobulins against several Mycobacterium leprae antigens have been indicated for the early diagnosis, and IgA participates in the early stages of leprosy and in subclinical infection, relatively little research has examined anti-M. leprae IgA responses. Here, we investigated serum IgA reactivity against NDO-HSA, LID-1 and NDO-LID, in paucibacillary (PB) and multibacillary (MB) patients and their household contacts, using enzyme-linked immunosorbent assay (ELISA). Diagnostic accuracy of each ELISA was evaluated by receiver operating characteristic (ROC) curve analysis. Our data reveal elevated IgA serum levels against the three M. leprae specific antigens in MB patients, whereas IgA reactivity in PB patients was increased only to NDO-HSA. Further, MB and PB household contacts displayed higher IgA reactivity to NDO-HSA than non-endemic controls. Our data suggest measurement of serum IgA against NDO-HSA as an additional tool in the diagnosis and classification of the disease, with potential utility for household contact follow-up.

Anatomia radiológica vascular e desenvolvimento de plataformas de ensino interativo / Vascular radiological anatomy and the development of interactive teaching platforms

Anatomia e Radiologia são disciplinas amplamente presentes nas grades curriculares de cursos da área da saúde. A interseção entre a Anatomia Radiológica e as tecnologias recentes, como plataformas de ensino interativo, caracteriza-se como uma tendência a ser seguida na esfera educacional, sendo as implicações desse processo ainda não muito debatidas. O Departamento de Anatomia da Universidade Federal de Juiz de Fora iniciou o desenvolvimento de um atlas de anatomia vascular radiológica, o qual apresenta versões impressa e interativa. Possui imagens de angiotomografias computadorizadas feitas em aparelhos multislice de 16 a 128 canais, sendo as imagens de arquivo pessoal dos organizadores. O atlas promove a identificação das estruturas vasculares em questão e propicia a integração do conhecimento adquirido em salas de aula com a visualização de exames de imagem complementares rotineiros. A indiscutível presença de exames de imagem adicionais na rotina dos profissionais da saúde atuais faz com que seja válida a estratégia de integração entre o método tradicional de ensino e as práticas ativas de aprendizagem, as quais vêm ganhando destaque recentemente, devido a sua efetividade na fixação de conhecimentos.

Anatomy and Radiology are subjects widely present in health courses at universities. The intersection between Radiological Anatomy and recent technologies, such as interactive teaching platforms, is characterized as a tendency to be followed in the educational sphere, and the implications of this process are not much debated. The Department of Anatomy of Juiz de Fora Federal University has begun the development of an atlas of radiological vascular anatomy, which features printed and interactived versions. It presents computed tomography images taken on multislice devices from 16 to 128 channels, and the images are from the personal authors' archives. The Radiologic Vascular Atlas provides the identification of vascular structures in study and makes easier the integration of knowledge acquired in classrooms with the visualization of routine complementary imaging exams. The indisputable presence of complementary imaging examinations in the routine of current health professionals makes valid the strategy of integration between the traditional teaching method and active learning practices, which have recently gained prominence, due to its effectiveness in setting knowledge.

Performance of recombinant proteins in diagnosis and differentiation of canine visceral leishmaniasis infected and vaccinated dogs.

Control of canine visceral leishmaniasis (CVL), a major zoonotic disease in Brazil and many other tropical and subtropical countries, remains difficult as an accurate and reliable diagnosis is still missing. In endemic regions, infected dogs are the main parasitic reservoir host of human Visceral leishmaniasis (VL) infection. Vaccination of dogs against Leishmania infection constitutes an important strategy to prevent or to better control CVL, thus, a serological test that can discriminate between antibodies induced by immunization versus infection is highly desirable in order to improve and simplify diagnosis. Here, four recombinant proteins were evaluated for their ability to detect and differentiate between dogs that are infected with Leishmania or have been immunized with the anti-Leishmania vaccine Leish-Tec®. Receiver operating characteristic (ROC) curve analysis of the four Leishmania-specific IgG ELISA revealed superior performance of rK28, followed by rKLO8, rK39 and rLb6H. The rK28-based ELISA revealed not only the best accuracy against CVL, but also the lowest cross-reactivity with sera from Leish-Tec® immunized dogs. Our data show that the rK28-based ELISA is highly suitable for CVL screening as it shows high sensitivity with simultaneous low cross-reactivity. Further, the high specificity of the rKLO8 indicates its suitability for the confirmation of CVL diagnosis.

Epidemiological and clinical assessment of patients undergoing Videolaparoscopic Cholecystectomy at a Curitiba teaching hospital. / Análise do perfil clínico epidemiológico dos pacientes submetidos a Colecistectomia Videolaparoscópica em um hospital de ensino de Curitiba.

OBJECTIVE: to assess clinical and epidemiological factors of patients undergoing video cholecystectomy at the Hospital da Cruz Vermelha do Paraná, Curitiba unit, operated from September 2016 to September 2018, as well as the influence of comorbidities, sex and advanced age on the postoperative prognosis, while in the hospital. METHODS: Analytic retrospective study. The analyzed variables were obtained by the review of medical records. Statistical analyses were performed considering the significance level p < 0.05. RESULTS: 389 patients, of whom 265 were women and 124 men, were included. The mean age was 51.5 years, 58.8% of the patients were diagnosed with at least one comorbidity, and 74.6% were overweight or obese. The incidence of intraoperative complications was 1.3%, postoperative 3.8% and, mortality, 0.3%. Some risk factors were identified as worse postoperative prognosis, such as Diabetes Mellitus, hypertension, presence of one or more comorbidities and mainly, advanced age, which was related to longer hospitalization times (p < 0,001), need of intensive therapy (p < 0,001), conversion to open surgery (p = 0,003) and postoperative complications (p < 0,001). Furthermore, the male sex was predictive of longer hospitalization times (p = 0,003) and need of intensive therapy (p = 0,01). CONCLUSION: the presence of comorbidities, male sex, and advanced age are predictive factors of bad prognosis for patients undergoing video laparoscopic cholecystectomy.

OBJETIVO: Analisar o perfil clínico e epidemiológico de pacientes submetidos a colecistectomia por vídeo no Hospital da Cruz Vermelha do Paraná, unidade de Curitiba, operados no período de setembro de 2016 a setembro de 2018, assim como a influência de comorbidades, sexo e idade avançada no prognóstico pós-operatório, durante o mesmo internamento. MÉTODOS: Estudo retrospectivo analítico, com dados coletados por revisão de prontuários. Análises estatísticas realizadas, considerando nível de significância p < 0,05. RESULTADOS: 389 casos foram incluídos, sendo 265 mulheres e 124 homens. A média de idade foi de 51,5 anos, 58,8% dos pacientes eram portadores de pelo menos uma comorbidade e 74,6% dos doentes tinham sobrepeso ou eram obesos. A incidência de complicações intraoperatórias foi de 1,3%, pós-operatórias de 3,8% e, a mortalidade, de 0,3%. Foram identificados fatores clínicos relacionados a pior prognóstico pós-operatório, como DM, HAS, presença de uma ou mais comorbidades e, destacadamente, idade avançada, que relacionou-se com maior tempo de internamento (p < 0,001), necessidade de UTI (p < 0,001), conversão para operação aberta (p = 0,003) e risco de complicações pós-operatórias (p < 0,001). Além disso, o sexo masculino foi preditivo para maior tempo de internamento (p = 0,003) e maior necessidade de UTI (p = 0,01). CONCLUSÃO: o perfil clínico-epidemiológico corresponde àquele exposto na literatura e a presença de comorbidades, o sexo masculino e a idade avançada são fatores preditivos de mau prognóstico na evolução pós-operatória de pacientes submetidos a colecistectomia videolaparoscópica.

Análise do perfil clínico epidemiológico dos pacientes submetidos a Colecistectomia Videolaparoscópica em um hospital de ensino de Curitiba / Epidemiological and clinical assessment of patients undergoing Videolaparoscopic Cholecystectomy at a Curitiba teaching hospital

RESUMO Objetivo: Analisar o perfil clínico e epidemiológico de pacientes submetidos a colecistectomia por vídeo no Hospital da Cruz Vermelha do Paraná, unidade de Curitiba, operados no período de setembro de 2016 a setembro de 2018, assim como a influência de comorbidades, sexo e idade avançada no prognóstico pós-operatório, durante o mesmo internamento. Métodos: Estudo retrospectivo analítico, com dados coletados por revisão de prontuários. Análises estatísticas realizadas, considerando nível de significância p < 0,05. Resultados: 389 casos foram incluídos, sendo 265 mulheres e 124 homens. A média de idade foi de 51,5 anos, 58,8% dos pacientes eram portadores de pelo menos uma comorbidade e 74,6% dos doentes tinham sobrepeso ou eram obesos. A incidência de complicações intraoperatórias foi de 1,3%, pós-operatórias de 3,8% e, a mortalidade, de 0,3%. Foram identificados fatores clínicos relacionados a pior prognóstico pós-operatório, como DM, HAS, presença de uma ou mais comorbidades e, destacadamente, idade avançada, que relacionou-se com maior tempo de internamento (p < 0,001), necessidade de UTI (p < 0,001), conversão para operação aberta (p = 0,003) e risco de complicações pós-operatórias (p < 0,001). Além disso, o sexo masculino foi preditivo para maior tempo de internamento (p = 0,003) e maior necessidade de UTI (p = 0,01). Conclusão: o perfil clínico-epidemiológico corresponde àquele exposto na literatura e a presença de comorbidades, o sexo masculino e a idade avançada são fatores preditivos de mau prognóstico na evolução pós-operatória de pacientes submetidos a colecistectomia videolaparoscópica.

ABSTRACT Objective: to assess clinical and epidemiological factors of patients undergoing video cholecystectomy at the Hospital da Cruz Vermelha do Paraná, Curitiba unit, operated from September 2016 to September 2018, as well as the influence of comorbidities, sex and advanced age on the postoperative prognosis, while in the hospital. Methods: Analytic retrospective study. The analyzed variables were obtained by the review of medical records. Statistical analyses were performed considering the significance level p < 0.05. Results: 389 patients, of whom 265 were women and 124 men, were included. The mean age was 51.5 years, 58.8% of the patients were diagnosed with at least one comorbidity, and 74.6% were overweight or obese. The incidence of intraoperative complications was 1.3%, postoperative 3.8% and, mortality, 0.3%. Some risk factors were identified as worse postoperative prognosis, such as Diabetes Mellitus, hypertension, presence of one or more comorbidities and mainly, advanced age, which was related to longer hospitalization times (p < 0,001), need of intensive therapy (p < 0,001), conversion to open surgery (p = 0,003) and postoperative complications (p < 0,001). Furthermore, the male sex was predictive of longer hospitalization times (p = 0,003) and need of intensive therapy (p = 0,01). Conclusion: the presence of comorbidities, male sex, and advanced age are predictive factors of bad prognosis for patients undergoing video laparoscopic cholecystectomy.

Proteínas de checkpoint imunológico como novo alvo da imunoterapia contra o câncer: revisão da literatura / Immune checkpoint proteins as new target for cancer immunotherapy: literature review

Uma nova era no tratamento do câncer está surgindo com o uso de anticorpos capazes de inibir pontos de bloqueio do sistema imunológico, chamados de "inibidores de checkpoint". Um novo conceito de "balas mágicas", que no início do século passado foram imaginadas por Paul Ehrlich como capazes de atuar diretamente na destruição de alvos tumorais, é representado agora por anticorpos direcionadas contra moléculas que bloqueiam a atividade antitumoral do sistema imunológico, como o antígeno-4 de linfócito T citotóxico (CTLA-4) e a proteína-1 de morte celular programada (PD-1). Essas novas imunoterapias vêm revolucionando a forma de tratar diferentes tipos de câncer. Nesta revisão selecionamos estudos sobre CTLA-4 e PD-1, seus ligantes em células apresentadoras de antígenos, assim como destacamos a importância da descoberta de antígenos tumorais e o papel do sistema imunológico na imunovigilância tumoral. Nesse estudo são discutidos aspectos relacionados aos efeitos de imunoterapias baseadas no uso de anticorpos monoclonais anti-CTLA-4 e anti-PD-1/ PD-L1, como o risco de serem estimuladas respostas direcionadas a tecidos saudáveis e outros efeitos adversos, bem como o uso de terapias combinadas que podem contribuir para melhorar a eficiência do tratamento do câncer.

A new era in cancer treatment is emerging with the use of antibodies directed against immune checkpoint proteins, known as "checkpoint inhibitors". A novel concept of "magic bullets", concepted by Paul Ehrlich at the beginning of the last century, as being capable of acting directly on the destruction of tumor targets, is now represented by antibodies directed against molecules which block the antitumor activity of the immune system, such as Cytotoxic T Lymphocyte Antigen-4 (CTLA-4) and Programmed Cell Death Protein-1 (PD-1). These new immunotherapies have revolutionized the treatment of different cancer types. Studies on CTLA-4, PD-1, and their ligands in antigen presenting cells are discussed in this review. The importance of tumor antigen discovery and the role of the immune system in immune surveillance of tumors were highlighted. Also in the present study, aspects related to the effects of immunotherapies based on the use of anti-CTLA-4 and anti-PD-1/ PD-L1 monoclonal antibodies are described, such as the risk of stimulating responses to normal tissues and other adverse effects, as well as the use of combination therapies which can improve the efficacy of cancer treatment.

Utility of immunoglobulin isotypes against LID-1 and NDO-LID for, particularly IgG1, confirming the diagnosis of multibacillary leprosy.

BACKGROUND Leprosy remains a health problem in many countries, with difficulties in diagnosis resulting in delayed treatment and more severe disabilities. Antibodies against several Mycobacterium leprae antigens have, however, shown value as diagnostic and/or prognostic markers. OBJECTIVES The objective of this study was to evaluate serum immunoglobulin (Ig) IgM and IgG subclass reactivity against three M. leprae specific antigens: NDO-HSA, a conjugate formed by natural octyl disaccharide bound to human serum albumin; LID-1, the fusion protein product of the ml0405 and ml2331 genes; and NDO-LID, a combination of LID-1 and NDO. METHODS Sera from healthy controls, paucibacillary (PB) and multibacillary (MB) leprosy patients, and their respective household contacts, were evaluated for the presence of antigen-specific IgM, IgG, and IgG subclass antibodies by enzyme-linked immunosorbent assay (ELISA). The sensitivity and specificity of each ELISA were evaluated by receiver operating characteristic (ROC) curve analysis. FINDINGS Our data confirm that serum IgM antibodies against NDO-HSA and IgG antibodies against LID-1, as well as IgG/M antibodies against NDO-LID, are markedly increased in MB patients. For the first time, our data reveal a selective increase in IgG1 and IgG3 antibodies against LID-1 and NDO-LID in MB patients, demonstrating that these antibody isotypes are suitable for differentiation between MB and PB patients. ROC curve analysis indicates an improved capacity for diagnosing MB leprosy patients using the detection of IgG antibodies, particularly the IgG1 isotype, specific to LID-1 and NDO-LID over the performance levels attained with NDO-HSA. CONCLUSIONS Our findings indicate that serological tests based on the detection of antigen-specific IgG1 antibodies are a useful tool to differentiate MB from PB patients, and indicate the enhanced performance of the LID-1 and NDO-LID antigens in the serodiagnosis of leprosy.

Utility of immunoglobulin isotypes against LID-1 and NDO-LID for, particularly IgG1, confirming the diagnosis of multibacillary leprosy

BACKGROUND Leprosy remains a health problem in many countries, with difficulties in diagnosis resulting in delayed treatment and more severe disabilities. Antibodies against several Mycobacterium leprae antigens have, however, shown value as diagnostic and/or prognostic markers. OBJECTIVES The objective of this study was to evaluate serum immunoglobulin (Ig) IgM and IgG subclass reactivity against three M. leprae specific antigens: NDO-HSA, a conjugate formed by natural octyl disaccharide bound to human serum albumin; LID-1, the fusion protein product of the ml0405 and ml2331 genes; and NDO-LID, a combination of LID-1 and NDO. METHODS Sera from healthy controls, paucibacillary (PB) and multibacillary (MB) leprosy patients, and their respective household contacts, were evaluated for the presence of antigen-specific IgM, IgG, and IgG subclass antibodies by enzyme-linked immunosorbent assay (ELISA). The sensitivity and specificity of each ELISA were evaluated by receiver operating characteristic (ROC) curve analysis. FINDINGS Our data confirm that serum IgM antibodies against NDO-HSA and IgG antibodies against LID-1, as well as IgG/M antibodies against NDO-LID, are markedly increased in MB patients. For the first time, our data reveal a selective increase in IgG1 and IgG3 antibodies against LID-1 and NDO-LID in MB patients, demonstrating that these antibody isotypes are suitable for differentiation between MB and PB patients. ROC curve analysis indicates an improved capacity for diagnosing MB leprosy patients using the detection of IgG antibodies, particularly the IgG1 isotype, specific to LID-1 and NDO-LID over the performance levels attained with NDO-HSA. CONCLUSIONS Our findings indicate that serological tests based on the detection of antigen-specific IgG1 antibodies are a useful tool to differentiate MB from PB patients, and indicate the enhanced performance of the LID-1 and NDO-LID antigens in the serodiagnosis of leprosy.

Testes sorológicos anti-NDO-HSA, anti-LID-1 e anti-NDO-LID em contatos domiciliares de área não endêmica de hanseníase / Anti-NDO-HSA, anti-LID-1 and anti-NDO-LID serological tests in household contacts of a non-endemic leprosy area

Introdução: Diversos fatores podem interferir no desenvolvimento da hanseníase, entre eles fatores genéticos, convívio com o caso de hanseníase e classificação operacional do caso. Testes sorológicos que avaliam a reatividade de anticorpos IgM e IgG frente a antígenos específicos para o Mycobacterium leprae (M. leprae) podem atuar como auxiliares na vigilância dos contatos e/ou população de risco. Objetivo: Analisar o comportamento dos testes sorológicos anti-PGL-1 sintético (NDO-HSA), anti-LID-1 e anti-NDO-LID em área não endêmica de hanseníase e sua relação com características do caso de hanseníase. Material e métodos: Trata-se de um estudo transversal, do tipo analítico, realizado com 35 contatos domiciliares (CD) dos casos de hanseníase. A coleta de dados ocorreu no período de agosto/2016 a fevereiro/2017 por meio de visitas domiciliares. A reatividade de anticorpos IgM e IgG frente aos antígenos Natural disaccharide linked to human serum albumin via octyl (NDO-HSA), Leprosy IDRI diagnostic 1 (LID-1) e Natural disaccharideoctyl - Leprosy IDRI Diagnostic 1(NDO-LID) foi avaliada por ensaio imunoenzimático (ELISA). Os dados foram exportados e analisados no software StatisticalPackage for the Social Sciences (SPSS®) 24 for Windows. Resultados: Foi observada maior proporção de positividade aos testes em CD de casos multibacilares (MB), que residiam com o caso de hanseníase na época do diagnóstico e que tinham parentesco consanguíneo com o caso. Esses casos de hanseníase MB também apresentaram soropositividade frente aos antígenos testados. O valor do índice ELISA foi maior no grupo de CD de casos MB. Houve concordância moderada e significativa (K= 0,53; p< 0,0001) entre os testes anti-NDO-HSA e anti-NDO-LID, mas não foi detectada diferença entre os testes anti-NDO-HSA e anti-LID-1 (K= -0,05; p= 0,678). A correlação foi positiva entre os três antígenos, porém, entre LID-1 e NDO-HSA, não houve significância estatística (p<0,186). Conclusão: Os resultados sugerem que testes sorológicos em conjunto com as características avaliadas nos contatos domiciliares em área não endêmica de hanseníase,podem atuar como auxiliares na detecção de indivíduos infectados pelo M. leprae, contribuindo para vigilância dos contatos domiciliares

Introduction: Several factors may interfere in the development of leprosy, including genetic factors, conviviality with leprosy patients and operational classification of the case. Serological tests performed to evaluate the reactivity of IgM and IgG antibodies response against Mycobacterium leprae (M. leprae) specific antigens may be used as auxiliary tools for transmission surveillance and/or population at risk. Objective: To analyze the performance of anti-PGL-1 (NDO-HSA), anti-LID-1 and anti-NDO-LID serological tests in non-endemic area of leprosy and the relationship with characteristics of the leprosy case. Material and methods: This is a cross-sectional analytical study of 35 household contacts (HC) of leprosy cases. Data collection was carried out from August 2016 to February 2017 with home visits. The reactivity of IgM and IgG antibodies to Natural disaccharide linked to human serum albumin via octyl (NDO-HSA), Leprosy IDRI diagnostic 1 (LID-1) and Natural disaccharide octyl - Leprosy IDRI Diagnostic 1 (NDO-LID) was evaluated through enzyme-linked immunosorbent assay (ELISA). Data were exported and analyzed using Statistical Package for Social Sciences (SPSS®) 24 for Windows. Results: A higher proportion of positivity was observed in the HC tests of multibacillary (MB) leprosy cases who lived in the same dwelling with a leprosy case at the time of diagnosis and had a degree of kinship with the case. These multibacillary leprosy cases also showed seropositivity to the antigens tests. ELISA test index value was higher in the HC group of MB leprosy cases. There was moderate agreement (K = 0.53, p <0.0001) between anti-NDO-HSA and anti-NDO-LID tests, but no difference was found between anti-NDO-HSA and anti-LID -1 (K = -0.05, p = 0.678). Three antigens were positively correlated, but there was no statistical significance (p <0.186) between LID-1 and NDO-HSA. Conclusion: The results suggest that serological tests in combination with the characteristics assessed during household contacts in a non-endemic area may represent efficient auxiliary tools for the detection of M. leprae-infected individuals, providing a contribution to the surveillance of household contacts

Efeitos contrastantes da vitamina D sobre a resposta imune inata e adquirida e seu impacto na recuperação da tuberculose / Contrasting effects of Vitamin D on innate and acquired immune responses and its impact on tuberculosis recover

A vitamina D é um hormônio essencial para o organismo, podendo ser obtida da dieta ou, principalmente, gerada pela pele após exposição à luz solar ultravioleta B. Na sua forma ativa (1,25(oH)2D) ela controla a absorção de cálcio e fósforo do intestino para a corrente sanguínea e participa de diversos processos celulares e fisiológicos. A ligação da 1,25(oH)2D ao receptor da vitamina D (VDr) presente em diversas células, como as células do sistema imunológico, induz a transcrição de genes que podem, por exemplo, modular a resposta imune inata e adquirida. A deficiência de vitamina D ou do VDR é associada a problemas de saúde como desordens esqueléticas, hipertensão, doenças cardiovasculares, diabetes mellitus, dislipidemias, doenças autoimunes e doenças infecciosas. Neste sentido, a suplementação com vitamina D tem sido proposta como uma possível medida preventiva, podendo ser aplicada em muitas patologias, em especial na tuberculose. Principal causa de morte por um único agente infeccioso, a tuberculose é responsável por cerca de 1,3 milhões de óbitos por ano no mundo. Publicações recentes apontam efeitos diversos da vitamina D na resposta imune inata e adquirida. A 1,25(oH)2D3 na presença do interferon (IFN)-γ é capaz de aumentar a atividade bactericida do macrófago contra o M. tuberculosis, aumentando a produção de peptídios antimicrobianos e estimulando a autofagia, favorecendo assim a lise de bacilos localizados em fagossomos. Por outro lado, a vitamina D em linfócitos T mostra efeito tolerogênico que favorece o controle de respostas inflamatórias excessivas. Neste trabalho de revisão são apresentados estudos recentes envolvendo efeitos da vitamina D na resposta imune inata e adquirida. Além disso, considerações sobre deficiência de vitamina D e maior risco de contrair tuberculose, e efeitos contrastantes da suplementação com vitamina D na prevenção e tratamento da TB, são discutidos.

Vitamin D is an essential hormone for the body, and can be obtained from diet or, mainly, generated by the skin after exposure to ultraviolet B sunlight. In its active form (1.25(oH)2D) it controls the absorption of calcium and phosphorus from the intestine into the bloodstream and participates in several cellular and physiological processes. Binding of 1,25(oH)2D to the Vitamin D receptor (VDr) present in several cells, such as cells of the immune system, induces transcription of genes that can, for example, modulate the innate and adaptive immune response. Deficiency of Vitamin D or VDr is associated with health problems such as skeletal disorders, hypertension, cardiovascular disease, diabetes mellitus, dyslipidemias, autoimmune diseases and infectious diseases. In this sense, Vitamin D supplementation has been proposed as a possible preventive measure and can be applied in several pathologies, especially in tuberculosis. main cause of death by a single infectious agent, tuberculosis accounts for about 1.3 million deaths per year worldwide. recent publications point to contrasting functions of Vitamin D in the innate and acquired immune response. 1.25(oH)2D3 in the presence of interferon (IFN)-γ is capable of increasing the bactericidal activity of the macrophage against M. tuberculosis, increasing the production of antimicrobial peptides and stimulating autophagy, thus favoring the lysis of bacilli located in phagosomes. on the other hand, Vitamin D in T lymphocytes shows a tolerogenic effect that favors the control of excessive inflammatory responses. In this review, recent studies involving Vitamin D effects on the innate and acquired immune responses are presented. In addition, considerations about Vitamin D deficiency and increased risk of contracting tuberculosis, and contrasting effects of Vitamin D supplementation on the prevention and treatment of TB, are discussed.

Diagnostic accuracy of rKLO8 versus rK26 ELISAs for screening of canine visceral leishmaniasis.

Canine visceral leishmaniasis (CVL) represents an important public health issue. Despite numerous diagnostic tests available, CVL diagnosis still needs to be improved to achieve a more accurate detection rate. Recently, rKLO8, a new antigenic protein of Sudanese Leishmania donovani, was studied for the first time in diagnosis of human visceral leishmaniasis (HVL) and showed good performance. The present study aimed to evaluate serum reactivity to rKL08 and the reference antigen rK26, and to compare both diagnostic proteins with the combined DPP® CVL rapid test and ELISA (EIE-Bio-Manguinhos) confirmatory test, which are both recommended for the diagnosis of CVL in Brazil. Serum samples of dogs were grouped into: (I) DPP®/EIE negative (n=100) and (II) DPP®/EIE positive sera (n=100). Enhanced levels of IgG, mainly IgG2, to both rKLO8 and rK26 were found in group II. Sensitivity was 68% and 77% and specificity was 92% and 91%, for rKLO8 and rK26 antigens, respectively. Moreover, the combination of rKLO8 and rK26 antigens (rKLO8+rK26) exhibited higher sensitivity (85%) and specificity (93%). Thus, our results show that apart from the improved diagnostic power of rKLO8 in HVL, this new antigen is also suitable for the diagnosis of CVL. Further, the combination of rKLO8 and rK26 antigens increases the diagnostic accuracy of CVL.

CCL2, CXCL8, CXCL9 and CXCL10 serum levels increase with age but are not altered by treatment with hydroxychloroquine in patients with osteoarthritis of the knees.

AIM: Osteoarthritis (OA) is a major cause of morbidity and incapacity in the elderly. This study evaluates serum levels of the chemokines CCL2, CXCL8, CXCL9, and CXCL10 in 16 patients with primary OA of the knees, and investigates how treatment with hydroxychloroquine (HCQ) for 4 months affects these chemokine levels. METHOD: Thirteen elderly patients received a placebo. Healthy control groups consisted of 10 elderly individuals (age > 60 years) with no clinical or radiological evidence of OA (CT-O), and 10 young adult individuals, (CT-Y group, age < 40 years). RESULTS: The CT-Y group presented lower levels of all chemokines studied, in comparison to the other groups. HCQ treatment did not alter the serum levels of CCL2 (P = 0.80), CXCL8 (P = 0.76), CXCL9 (P = 0.95) and CXCL10 (P = 0.74) in OA patients. CONCLUSION: Hydroxychloroquine treatment did not alter the serum levels of CCL2, CXCL8, CXCL9 or CXCL10 in patients with OA of the knees, although increased serum levels correlated with aging for all subjects, including controls.

Autotransplantation of Spleen Mitigates Drug-Induced Liver Damage in Splenectomized Mice.

PURPOSE: The spleen presents numerous functions, including the production of immunoglobulins and blood filtration, removing microorganisms and cellular debris. The spleen also has anatomical and functional relationship with the liver, but there are few studies on this topic. The aim of this study was to assess the effect of splenectomy and autologous spleen transplantation on both filtering functions of spleen and acetaminophen-induced hepatotoxicity. MATERIALS AND METHODS: Fifty-two BALB/c mice were randomized into four groups: splenectomized; splenectomy and splenic autotransplantation in the greater omentum; sham operated control; and non-operated control. At day 7th, 14th, and 28th after surgery, splenic filtration was assessed by counting Howell-Jolly bodies (HJB) and pitted red cells (PIT). The animals received 400 mg/kg acetaminophen by gavage at day 28th and after 12 or 24 hours were euthanized for evaluation of splenic and hepatic morphology. RESULTS: The splenectomized group demonstrated reduced filtration of HJB and PIT in all analyzes, while the autotransplanted group developed progressive recovery of function after the 14th day. At day 28 after surgery the implants showed similar histology in comparison to normal spleen. Liver histology showed more intense centrilobular necrosis in splenectomized group in comparison to the others, suggesting a protective role of spleen in acetaminophen-induced liver injury. CONCLUSIONS: Splenic implants showed structural and functional recovery, demonstrating the ability of autologous implant to rescue filtering function of intact spleen. Furthermore, the integrity of splenic function appears to influence liver morphology, since the presence of the splenic implants mitigated the effects of chemically-induced liver damage.

Detection of IgG1 antibodies against Mycobacterium tuberculosis DosR and Rpf antigens in tuberculosis patients before and after chemotherapy.

Diagnosis of tuberculosis (TB) remains challenging. Serum IgG1 antibodies against Mycobacterium tuberculosis active growth phase antigens (ESAT-6/CFP-10, Rv0717 and Rv3353), DosR regulon-encoded proteins (Rv1733, Rv1737, Rv2628 and Rv2029), and resuscitation-promoting factors (Rv0867 and Rv2389) were evaluated in TB patients using ELISA. Active TB patients showed elevated levels of IgG1 antibodies against ESAT-6/CFP-10, Rv0717, Rv3353, Rv1733, Rv2628, Rv2029 and Rv0867 in comparison to healthy controls (p < 0.001). These levels remained high after the initiation of treatment, while responses to Rv0717 and Rv1733 peaked early during treatment. IgG1 responses to ESAT-6/CFP-10, Rv3353, Rv2628, Rv2029 and Rv0867 declined to control levels after the completion of 6 months chemotherapy. ROC analysis confirmed the good diagnostic performance of Rv0717, Rv1733, Rv3353, Rv2628, Rv2029 and Rv0867antigens. These data suggest that detecting IgG1 antibodies against M. tuberculosis antigens, including DosR and Rpf proteins, may represent an additional tool in the diagnosis of tuberculosis.

Challenging Mycobacterium tuberculosi s dormancy mechanisms and their immunodiagnostic potential

ABSTRACT Mycobacterium tuberculosis is the etiologic agent of tuberculosis, one of the world's greatest cause of morbidity and mortality due to infectious disease. Many evolutionary mechanisms have contributed to its high level of adaptation as a host pathogen. Prior to become dormant, a group of about 50 genes related to metabolic changes are transcribed by the DosR regulon, one of the most complex and important systems of host-pathogen interaction. This genetic mechanism allows the mycobacteria to persist during long time periods, establishing the so-called latent infection. Even in the presence of a competent immune response, the host cannot eliminate the pathogen, only managing to keep it surrounded by an unfavorable microenvironment for its growth. However, conditions such as immunosuppression may reestablish optimal conditions for bacterial growth, culminating in the onset of active disease. The interactions between the pathogen and its host are still not completely elucidated. Nonetheless, many studies are being carried out in order to clarify this complex relationship, thus creating new possibilities for patient approach and laboratory screening.

Challenging Mycobacterium tuberculosis dormancy mechanisms and their immunodiagnostic potential.

Mycobacterium tuberculosis is the etiologic agent of tuberculosis, one of the world's greatest cause of morbidity and mortality due to infectious disease. Many evolutionary mechanisms have contributed to its high level of adaptation as a host pathogen. Prior to become dormant, a group of about 50 genes related to metabolic changes are transcribed by the DosR regulon, one of the most complex and important systems of host-pathogen interaction. This genetic mechanism allows the mycobacteria to persist during long time periods, establishing the so-called latent infection. Even in the presence of a competent immune response, the host cannot eliminate the pathogen, only managing to keep it surrounded by an unfavorable microenvironment for its growth. However, conditions such as immunosuppression may reestablish optimal conditions for bacterial growth, culminating in the onset of active disease. The interactions between the pathogen and its host are still not completely elucidated. Nonetheless, many studies are being carried out in order to clarify this complex relationship, thus creating new possibilities for patient approach and laboratory screening.

Efficacy of (99m)Tc-Labeled Ceftizoxime in the Diagnosis of Subclinical Infections Associated with Titanium Implants in Rats.

BACKGROUND: Metal implants are used frequently in orthopedic procedures and the occurrence of subclinical low-virulence infection is difficult to diagnose. The objective of this study was to examine the hypothesis that peri-prosthetic subclinical infections may be diagnosed effectively in a murine model system using scintigraphic imaging with (99m)Tc-labeled ceftizoxime. METHODS: A sample population of 3-mo old Wistar rats (mean weight 327 g) was divided randomly into a control group (n=6), which received sterile implants, and an experimental group (n=6), which received implants contaminated with Staphylococcus aureus strain ATCC6538-P. Animals were anesthetized and femoral titanium implants were fixed beneath muscle tissue in left hind limbs. Three weeks after surgery, animals were injected with (99m)Tc-ceftizoxime solution (62.9 MBq) and scintigraphic images were obtained at 3.5 and 6.5 h after tracer injection. RESULTS: According to the scintigraphic images, the radiopharmaceutical showed affinity for the operated thigh areas of experimental animals but not for those of the control group. There was no difference between the control and experimental groups regarding the amount of radioactivity in the regions of interest measured at 3.5 h after injection of radiolabeled antibiotic, but the between-group difference determined at 6.5 h after treatment was statistically significant (p=0.026). Moreover, the level of radioactivity recorded in resected thigh tissues derived from experimental animals was greater than that of the control group (p=0.035). CONCLUSION: (99m)Tc-ceftizoxime scintigraphy can localize preferentially periprosthetic-infected areas adjacent to metal implants in a murine model. Furthermore, the radiolabeled antibiotic appears to be capable of detecting alterations in the micro-environment close to the implant and of reaching the bacteria attached to the implant surface.