L-glutamine supplementation reduced morphological damage in the renal glomerulus of rats with Walker-256 tumor

ABSTRACT Purpose: To evaluate the effects of the experimental subcutaneous Walker-256 tumor and L-glutamine supplementation, an antioxidant, on the glomerular morphology of rats. Methods: Twenty Wistar rats were distributed into four groups (n = 5): control (C); control treated with 2% L-glutamine (CG); rats with Walker-256 tumor (WT); and rats with Walker-256 tumor treated with 2% L-glutamine (WTG). Renal histological samples were submitted to periodic acid-Schiff and Masson's Trichrome staining to analyze glomerular density, morphometry of glomerular components and glomerulosclerosis; and to immunohistochemistry for fibroblast growth factor-2 (FGF-2). Results: WT showed 50% reduction in body mass gain and cachexia index > 10%, while WTG demonstrated reduction in cachexia (p < 0.05). WT revealed reduction of glomerular density, increase in the glomerular tuft area, mesangial area, matrix in the glomerular tuft, decrease in the urinary space and synechia, and consequently higher glomerulosclerosis (p < 0.05). L-glutamine supplementation in the WTG improved glomerular density, and reduced glomerular tuft area, urinary space, mesangial area, and glomerulosclerosis compared to WT(p < 0.05). WT showed higher collagen area and FGF-2 expression compared to C (p < 0.05). WTG presented lower collagen fibers and FGF-2 expression compared to WT (p < 0.05). Conclusions: L-glutamine supplementation reduced cachexia and was beneficial for glomerular morphology of the rats, as well as it reduced kidney damage and improved the remaining glomeruli morphology.

Curcuma longa extract protects against 5-fluorouracil-induced oral mucositis in hamsters

Abstract Curcumin, contained at Turmeric (Curcumalonga), can exert many beneficial pleiotropic activities in the gastrointestinal tract. This study evaluated the antioxidant and anti-inflammatory activity of C. longa on 5-fluorouracil (5-FU)-induced oral mucositis (OM) in hamsters. Phytochemical analysis of crude C. longa extract (CLE) was performed to detect the presence of curcumin by TLC and HPLC. Golden Syrian hamsters were orally pre-treated with CLE (5, 50, or 100mg/kg). Cheek pouch samples were subjected to macroscopic and histopathological evaluation. ELISA was performed to quantify the inflammatory cytokines IL-1ß and TNF-α. Superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) levels were assessed by ultraviolet-visible spectroscopy analysis. Behavior analysis was conducted by the open field test. Curcumin content in the CLE was 0.55%m/m ± 0.0161 (2.84%). The group treated with 5mg/kg CLE showed healing evidence with macroscopic absence of ulceration (p<0.05) and microscopic aspect of re-epithelialization, discrete inflammatory infiltrate and absence of edema. Treatment with 5mg/kg CLE significantly increased GSH levels, and reduced MDA levels and SOD activity (p˂0.05), and decreased IL-1ß (p˂0.05) and TNF-α (p˂0.01) levels. A significant reduction in walking distance, ambulation, speed, and rearing was observed for motor activity. Curcumin reduced oxidative stress, inflammation, and motor activity in hamsters with 5-FU-induced OM.

Granuloma piógeno en el dorso lingual: caso clínico / Pyogenic granuloma in the lingual dorsum: a clinical case

El granuloma piógeno es una lesión benigna en la cavidad bucal, no neoplásica. Es una entidad fre- cuentemente asociada a la expansión de los tejidos blandos de la cavidad bucal. Se presenta, con mayor frecuencia, en individuos del género femenino, con edades comprendidas entre la segunda y cuarta década de vida. El tratamiento es mediante escisión quirúrgica con un pequeño margen de seguridad, y los agentes irritantes deben extraerse concomitan- temente para la curación de la lesión. Este trabajo tuvo como objetivo informar un caso de granuloma piógeno en el dorso de la lengua en una paciente del género femenino de 69 años. Los autores destacaron la importancia del conocimiento de la patología bucal por parte del Cirujano Dentista para poder realizar un correcto diagnóstico diferencial de otras lesiones, con el fin de realizar el tratamiento adecuado (AU)

Pyogenic granuloma is a quite common non-neoplasic benign lesion in the oral cavity. It is one of the entities most frequently associated with the soft tissues' expansion of the oral cavity, specifically in females, in the age group between the second and the fourth decade of life. The treatment is by surgical excision with a small margin of safety. For the healing of the lesion irritants should be concomitantly removed. This study aimed to report a case of pyogenic granuloma on the lingual dorsum of a 69-year-old female patient. The authors concluded highlighting the importance of the knowledge of oral pathology by the Dental Surgeon, to perform a correct differential diagnosis of other lesions to perform the appropriate treatment (AU)

Qualidade de vida, sintomas de depressão e adesão ao tratamento em pacientes com trantorno depressivo maior / Quality of life, depression symptoms and adherence to treatment in patients with major depressive disorder / Calidad de vida, síntomas depresión y adhesión al tratamiento en pacientes con trastorno depresivo mayor

Introdução:Pacientes com depressão maior geralmente respondem ao tratamento com medicamentos antidepressivos, no entanto em 10% a 30% dos casos há apenas uma resposta parcial ou nenhuma resposta, entre os fatores que podem influenciar encontra-se o perfil das enzimas hepáticas metabolizadoras dos antidepressivos, tal como a CYP2C19.Objetivo:Caracterizar os indivíduos quanto ao perfil genético dospolimorfismos CYP2C19*2 ou CYP2C19*17 em pacientes com transtorno depressivo maior (TDM) tratados com citalopram ou escitalopram e compará-los em relação a adesão ao tratamento, sintomas de depressão e qualidade de vida.Metodologia:Trata-se de um estudo transversal realizado com 29 pacientes com TDM. Amostras de sangue foram coletadas para genotipagem de CYP2C19 por discriminação alélica TaqMan®. Após caracterização do perfil genético, os indivíduos foram comparados quanto aos dados demográfico e socioeconômico, adesão ao tratamento (TestedeMorisky-Green),sintomas de depressão (escala de Hamilton) e qualidade de vida (WHOQoL-BREF).Resultados:Quatro pacientes (13.8%) apresentaram polimorfismo para CYP2C19*2 e 10 pacientes (34.4%) para CYP2C19*17, com maior prevalência de CYP2C19*17 (p>0.05). Nenhuma associação significativa de características socioeconômicas, demográficas e clínicas entre os genótipos do CYP2C19.No TestedeMorisky-Green, aadesão moderada ao tratamento foi predominante nos pacientes CYP2C19*2 e CYP2C19*17 (p>0.05). Não foi observada associação entre sintomas de depressão e polimorfismos genéticos (p>0.05). Uma associação significativa entre o genótipo polimórfico CC do CYP2C19*17 com a satisfação com a saúde, enquanto o genótipo CT foi associado ao estado "nem satisfeito/nem insatisfeito" (p<0.05). A maioria dos indivíduos CYP2C19*2 e CYP2C19*17 relatou "necessidade de melhorar" em relação aos domínios de qualidade de vida físico, psicológico, social e ambiental (p>0.05).Conclusões:Os pacientes apresentaram maior prevalência do polimorfismo CYP2C19*17, com moderada adesão ao tratamento. Alguns pacientes, mesmo sob efeito da medicação, apresentaram sintomas de depressão moderado a intenso e relataram uma indefinição na satisfação da sua qualidade de vida (AU).

Introduction:Patients with major depression usually respond to treatment with antidepressant drugs, however in 10% to 30% of cases there is only a partial response or no response, among the factors that can influence is the profile of liver enzymes metabolizing antidepressants, such as CYP2C19.Objective:To characterize the individuals regarding the genetic profile ofCYP2C19*2or CYP2C19*17 polymorphisms in patients with major depressive disorder (MDD) treated with citalopram or escitalopram, and to compare themaccording to treatment adherence, symptoms of depression and quality of life.Methodology:This is cross-sectionalstudy carried out with 29 patients with MDD. Blood samples were collected for CYP2C19 genotyping by TaqMan® allelic discrimination. After characterization of the genetic profile, the individuals were compared regarding the demographic and socioeconomic data, treatment adherence (Morisky-GreenTest), symptoms of depression (Hamilton scale) and quality of life (WHOQoL-BREF).Results:Four patients showed (13.8%) CYP219*2 and 10 patients (34.4%) CYP219*17 polymorphisms.,withhigher prevalence of CYP219*17 (p>0.05). No association between socioeconomic, demographic, and clinical features with CYP2C19 genotypes was observed. In Morisky-GreenTest, moderate adherence to treatment was predominant for CYP2C19*2 and CYP219*17 patients (p>0.05). No statistically significant association was observed between symptoms of depression and genetic polymorphisms (p>0.05). A significant association between polymorphic CC genotype of CYP219*17 with health satisfaction, while the CT genotype was associated with "neither satisfied/nor dissatisfied" status (p<0.05). Most of the CYP2C19*2 and CYP2C19*17 subjects reported "need to improve" or "regular" regarding physical, psychological, social, and environmental domainsof quality of life(p>0.05).Conclusions:The patients showed a higher prevalence of CYP219*17 polymorphism, with moderate treatment adherence. Some subjects, even under the effect of the medication, presented moderate to intense symptoms of depression, and reported a lack of definition in the satisfaction of their quality of life (AU).

Introducción:Los pacientes con depresión mayor responder al tratamiento con antidepresivos, en 10% al 30% de los casos existe una respuesta parcial o nula, entre los factores que pueden influir se encuentra el perfil de enzimas hepáticas metabolizadoras de antidepresivos, como CYP2C19.Objetivo: Caracterizar a los individuos en cuanto al perfil genético depolimorfismos CYP2C19 *2 o CYP2C19 * 17 en pacientes con trastorno depresivo mayor (TDM) tratados con citalopram o escitalopram y compararlos en relaciónpara la adherencia al tratamiento, síntomas de depresión y la calidad de vida.Metodología: Estudio transversalcon 29 pacientes con TDM. Se recogieron muestras de sangre para la determinación del genotipo CYP2C19 mediante discriminación alélica TaqMan®, los individuos fueron comparados en cuanto a los datosdemográficosy socioeconómicos, adherencia (Prueba de Morisky-Green), síntomas de depresión (escala de Hamilton) y calidad de vida (WHOQoL-BREF).Resultados: Cuatro pacientes (13,8%) con polimorfismo CYP2C19*2 y 10 (34,4%) con CYP2C19 * 17,(p> 0,05). No existe una asociación significativa de las características socioeconómicas, demográficas y clínicas con los genotipos CYP2C19. La adherencia moderada al tratamiento fue predominante en los pacientes con CYP2C19*2 y CYP2C19*17 (p> 0,05). No hubo asociación entre síntomas de depresión y polimorfismos genéticos (p> 0.05). Una asociación significativa entre el genotipo polimórfico CYP2C19 * 17 CC con la satisfacción con la salud, mientras que el genotipo CT se asoció con el estado "ni satisfecho / no insatisfecho" (p <0.05). La mayoría de CYP2C19 * 2 y CYP2C19 * 17 individuos informaron "necesidad de mejorar" en relación con los dominios físico, psicológico, social y ambientalde calidad de vida(p> 0,05).Conclusiones: Los pacients mostraron una mayor prevalencia del CYP2C19 * 17, con adherencia moderada al tratamiento, síntomas de depresión moderada a intensay informaron una falta de definición en la satisfacción de su calidad de vida (AU).

ABORDAGEM DAS TEMÁTICAS "SAÚDE BUCAL DE GESTANTES E BEBÊS" E "SAÚDE DO HOMEM" EM SALAS DE ESPERA: SIGNIFICÂNCIAS POLÍTICO-PEDAGÓGICAS DAS EXPERIMENTAÇÕES VIVENCIADAS EM UM ESTÁGIO SUPERVISIONADO / APPROACH TO THEMES "BUCAL HEALTH OF GESTANTS AND BABIES" AND "MAN'S HEALTH" IN WAITING ROOMS: POLITICAL-PEDAGOGICAL MEANINGS OF THE EXPERIMENTS ENCOURAGED IN A SUPERVISED STAGE

Objetivo: analisar as significâncias político-pedagógicas de duas vivências experimentadas ("Saúde bucal de gestantes e bebês" e "Saúde do homem") por estagiários de um curso de Odontologia em salas de espera de uma Unidade de Atenção Primária de Saúde (UAPS). Metodologia: estudo qualitativo transversalmente estruturado sob estratégia narrativo-descritiva e moldado à técnica argumentativa. Resultados: o "Estágio Supervisionado em Unidade de Atenção Primária I" foi didaticamente sistematizado em dois períodos, "Pré-intervenção" e "Intervenção". Do primeiro, se desvendaram três ações, a "Pactualização do enlace ensino-serviço-comunidade", a "Contextualização dos acadêmicos estagiários" e a "Estruturação e ambientalização das equipes de trabalho". Já o segundo foi guiado pela lógica pedagógica do instrumento "TPC" (Teorizar-Praticar-Criticar), onde todas as ações programadas seguiram a lógica ativa do planejamento estratégico, ou seja, contextualizadas às realidades do cenário de prática, ou seja, salas de espera da Unidade de Atenção Primária de Saúde do bairro JK do município de Governador Valadares, MG. Conclusão: das experimentações vivenciadas algumas inferências merecem destaque: a concepção pedagógica de estratégias práticas de ensino pautadas na articulação ensino-serviço-comunidade; a escuta na identificação dos problemas/demandas a serem enfrentadas pela equipe estagiária; o reconhecimento do potencial dos ambientes de espera para a implantação de ações educativas; a importância de disseminar os aprendizados advindos de experimentações práticas de estágios.

Objective: to analyze the political-pedagogical significance of two experienced experiences ("Oral Health of Pregnant Women and Babies" and "Men's Health") by trainees of a Dentistry course in waiting rooms of a Primary Health Care Unit. Methodology: qualitative study transversally structured under a narrative-descriptive strategy and molded to the argumentative technique. Results: The "Supervised Internship in Primary Care Unit I" was systematized in two periods, "Pre-intervention" and "Intervention". From the first, three actions were unveiled, the "Pactualization of the teaching-service-community link", the "Contextualization of trainee academics" and the "Structuring and environmentalization of work teams". The second one was guided by the pedagogical logic of the instrument "TPC" (Theorizar-Praticar-Criticar), where all the actions programmed followed the active logic of strategic planning, that is, contextualized to the realities of the practice scenario, expects the Primary Health Care Unit of the JK district of the municipality of Governador Valadares, MG. Conclusion: from the weightings listed some inferences deserve to be highlighted: the pedagogical conception of practical teaching strategies based on the teaching-service-community articulation; listening in the identification of problems / demands to be faced by the trainee team; the recognition of the potential of waiting environments for the implementation of educational actions; the importance of disseminating the learning that comes from practical experimentation of internships.

Modification in clic4 expression is associated with p53, tgf-ß, tnf-α and myofibroblasts in lip carcinogenesis

Abstract Chloride intracellular channel-4 (CLIC4) is regulated by p53 and tumor necrosis factor-α (TNF-α), it is linked to the increase of transforming growth factor-β (TGF-β), and myofibroblastic differentiation in skin carcinogenesis. This study analyzed the immunoexpression of CLIC4, p53, TGF-β, TNF-α, and α-SMA in 50 actinic cheilitis (AC) and 50 lower lip squamous cell carcinoma (LLSCC). AC and LLSCC immunoexpression were categorized as score 1 (<5% positive cells), 2 (5-50%) or 3 (>50%). For CLIC4, nuclear and cytoplasmic immunostaining of epithelial cells was considered individually. For morphologic analysis, the World Health Organization criteria were used to epithelial dysplasia grade of ACs, and Bryne grading of malignancy system was applied for LLSCC. Higher nuclear CLIC4 (CLIC4n) and TGF-β were observed in ACs with low-risk of transformation, while cytoplasmic CLIC4 (CLIC4c), p53 and TNF-α were higher in the high-risk cases (p<0.05). In LLSCCs, CLIC4c was higher in cases with lymph node metastasis, advanced clinical stages, and histological high-grade malignancy. p53 expression was higher in high-grade LLSCCs, whereas TGF-β decreased as the clinical stage and morphological grade progressed (p<0.05). ACs showed an increased expression of CLIC4n and TGF-β, while CLIC4c and α-SMA were higher in LLSCCs (p<0.0001). Both lesions showed negative correlation between CLIC4n and CLIC4c, while in LLSCCs, negative correlation was also verified between CLIC4c and p53, as well as CLIC4c and TGF-β (p<0.05). Change of CLIC4 from the nucleus to cytoplasm and alterations in p53, TGF-β, TNF-α, and α-SMA expression are involved in lip carcinogenesis.

Resumo O canal intracelular de cloreto 4 (CLIC4) é regulado pela p53 e fator de necrose tumoral α (TNF-α) e está relacionado ao aumento do fator de crescimento transformador β (TGF-β) e na diferenciação miofibroblástica na carcinogênese cutânea. Este estudo analisou a imunoexpressão de CLIC4, p53, TGF-β, TNF-α e α-SMA em 50 queilites actínicas (QA) e 50 carcinomas de células escamosas de lábio inferior (CCELI). A imunoexpressão da QA e CCELI foram categorizadas em escore 1 (<5% de células positivas), 2 (5-50%) ou 3 (>50%). Para CLIC4, a imunomarcação nuclear e citoplasmática das células epiteliais foi considerada separadamente. Para análise morfológica, foram utilizados os critérios da Organização Mundial da Saúde para a gradação das displasias epiteliais nas QAs, e o sistema de gradação de malignidade de Bryne foi utilizado para os casos de CCELIs. Alta imunoexpressão de CLIC4 nuclear (CLIC4n) e TGF-β foi observada em QA de baixo risco de transformação, enquanto CLIC4 citoplasmática (CLIC4c), p53 e TNF-α foram elevadas nos casos de alto risco (p<0.05). No CCELI, a imunoexpressão de CLIC4c foi maior em caos com metástase linfonodal, estágio clínico avançado e alto grau histológico de malignidade. A expressão de p53 foi elevada em CCELI de alto grau, enquanto o TGF-β diminuiu à medida que o estádio clínico e o grau morfológico progrediram (p<0.05). QAs exibiram uma elevada expressão de CLIC4n e TGF-β, enquanto o CLIC4c e α-SMA foram elevados em CCELIs (p<0.0001). Ambas as lesões mostraram correlação negativa entre CLIC4n e CLIC4c, enquanto nos CCELIs, também se verificou correlação negativa entre CLIC4c e p53, assim como entre CLIC4c e TGF-β (p<0.05). Alteração do CLIC4 do núcleo para o citoplasma e alterações na expressão de p53, TGF-β, TNF-α, e α-SMA estão envolvidas na carcinogênese labial.

Detection of Drug Resistant Mycobacterium Tuberculosis Strains Using Kit SIRE Nitratase®: a Multicenter Study

Abstract (1) Background: The Commercial Kit SIRE Nitratase® PlastLabor, is a drug susceptibility test kit used to detect Mycobacterium tuberculosis resistance to first-line TB treatment drugs. The present study aimed at evaluating its performance in a multicenter study. (2) Methods: To determine its accuracy, the proportion methods in Lowenstein Jensen medium or the BACTECTMMGITTM960 system was used as a gold standard. (3) Results: The study revealed that the respective accuracies of the kit with 190 M. tuberculosis clinical isolates, using the proportion methods in Lowenstein Jensen medium or BACTECTMMGITTM960 system as a gold standard, were 93.9% and 94.6%, 96.9% and 94.6%, 98.0% and 97.8%, and 98.0% and 98.9%, for streptomycin, isoniazid, rifampicin, and ethambutol, respectively. (4) Conclusion: Thus, the kit can rapidly screen resistance to streptomycin, isoniazid, rifampicin, and ethambutol. Additionally, it does not require sophisticated equipment; hence, it can be easily used in the laboratories of low and middle income countries.

Clinical aspects in patients with pulmonary infection caused by mycobacteria of the Mycobacterium abscessus complex, in the Brazilian Amazon / Aspectos clínicos em pacientes com infecção pulmonar por micobactérias do complexo Mycobacterium abscessus na Amazônia brasileira

ABSTRACT Objective: To describe the clinical manifestations of patients with pulmonary infection caused by mycobacteria of the Mycobacterium abscessus complex (MABSC), and to compare these manifestations with those of patients infected with other nontuberculous mycobacteria (NTM). Methods: This was a retrospective cohort study involving 43 patients divided into two groups: the MABSC group, consisting of patients with pulmonary infection caused by MABSC (n = 17); and the NTM group, consisting of patients with pulmonary infection caused by NTM other than MABSC (n = 26). Patients were previously treated with a regimen of rifampin, isoniazid, pyrazinamide, and ethambutol before the diagnosis of NTM was confirmed by two culture-positive sputum samples. The nucleotide sequences of the hsp65, 16S rRNA, and/or rpoB genes were analyzed to identify the mycobacteria. Data were collected on demographic, clinical, and radiological characteristics, as well as on treatment responses and outcomes. Results: Loss of appetite was the only clinical manifestation that was significantly more common in the MABSC group than in the NTM group (p = 0.0306). The chance of having to use a second treatment regimen was almost 12 times higher in the MABSC group than in the NTM group. Treatment success was significantly higher in the NTM group than in the MABSC group (83.2% vs. 17.6%; p < 0.0001). The chance of recurrence was approximately 37 times higher in the MABSC group than in the NTM group. Conclusions: In the study sample, treatment response of pulmonary disease caused by MABSC was less favorable than that of pulmonary disease caused by other NTM.

RESUMO Objetivo: Descrever as manifestações clínicas de pacientes com infecção pulmonar por micobactérias do complexo Mycobacterium abscessus (CMA) e compará-las com as daqueles infectados com outras micobactérias não tuberculosas (MNT). Métodos: Estudo de coorte retrospectivo envolvendo 43 pacientes divididos em dois grupos: grupo CMA, com pacientes com infecção pulmonar por CMA (n = 17); e grupo MNT, com pacientes com infecção pulmonar por MNT que não CMA (n = 26). Os pacientes foram previamente tratados com o esquema rifampicina, isoniazida, pirazinamida e etambutol antes de o diagnóstico de MNT ser confirmado com culturas positivas em duas amostras de escarro diferentes. As sequências nucleotídicas dos genes hsp65, RNAr 16S e/ou rpoB foram analisadas para a identificação das micobactérias. Foram coletadas características demográficas, clínicas e radiológicas, assim como respostas terapêuticas e desfechos. Resultados: A única manifestação clínica significativamente mais frequente no grupo CMA que no grupo MNT foi hiporexia (p = 0,0306). A chance de haver a necessidade de utilização de um segundo esquema terapêutico foi quase 12 vezes maior no grupo CMA que no grupo MNT. O sucesso terapêutico foi significativamente maior no grupo MNT que no grupo CMA (83,2% vs. 17,6%; p < 0,0001). A chance de recidiva no grupo CMA foi aproximadamente 37 vezes maior que no grupo MNT. Conclusões: Na amostra estudada, a resposta terapêutica da doença pulmonar causada por CMA evoluiu de forma menos favorável do que naquela causada pelas demais MNT.

Resposta imune e mecanismos de evasão na carcinogênese de lábio: um estudo imunoistoquímico / Immune response and evasion mechanisms in lip carcinogenesis: an immunohistochemical study

A vigilância imunológica, principalmente mediada por linfócitos T CD8+ , reconhece e destrói células malignas ou alteradas. Contudo, através de estratégias imunossupressoras, como as vias de sinalização do ligante de morte celular programada-1 (PD-L1) e do antígeno leucocitário humano-G (HLA-G), estas células mutadas conseguem escapar da resposta imune antitumoral. Este estudo investigou a imunoexpressão de PD-L1, HLA-G, CD8 e granzima B (GrB) no microambiente de carcinomas de células escamosas (CCEs) de lábio (n = 40), de queilites actínicas (QAs; n = 55) e de mucosa labial saudável (MLS; n = 10). As amostras foram submetidas à técnica da imunoistoquímica e as análises das imunomarcações seguiram métodos semi-quantitativos (PD-L1 e HLA-G) e quantitativos (CD8 e GrB). A expressão das proteínas foi comparada entre os três grupos de amostras, bem como com parâmetros clinicopatológicos das lesões e sobrevida global dos pacientes com CCE de lábio. A correlação entre as proteínas e o tipo do microambiente tumoral de acordo com a presença de PD-L1 e CD8 também foram avaliados. Os testes estatísticos incluíram o exato de Fisher, Mann-Whitney, Kruskal-Wallis, correlação de Spearman e log-rank para comparação das curvas de sobrevida global construídas pelo método Kaplan-Meier. O nível de significância foi estabelecido em 5%. Os números de células CD8+ e GrB+ aumentaram progressivamente de MLS para CCEs de lábio, com QAs exibindo números intermediários (p < 0,01). A menor expressão dessas proteínas foi associada à metástase para linfonodos e tumores pobremente diferenciados (p < 0,05). A expressão de PD-L1 e HLA-G em células neoplásicas/ceratinócitos e estroma/tecido conjuntivo foi significativamente maior em CCEs de lábio e QAs, em comparação com MLSs (p < 0,05). PDL1 não foi significativamente associado aos aspectos clinicopatológicos das lesões. A maioria dos CCEs de lábio mostrou coexistência de células PD-L1+ e CD8+ (72,5%) no microambiente tumoral. A expressão de PD-L1 foi diretamente correlacionada à infiltração linfocítica CD8+ e GrB+ em CCEs de lábio (p < 0,05). A expressão das proteínas não foi associada com a sobrevida global dos pacientes com CCEs de lábio (p > 0,05). Nossos achados sugerem que as moléculas imunossupressoras PD-L1 e HLA-G estão consistentemente expressas desde QAs e se mantém até fases avançadas dos CCE de lábio. A correlação entre a expressão de PD-L1 e a expressão de CD8 e GrB nos carcinomas sugere que PD-L1 pode surgir como um mecanismo de escape frente a uma resposta antitumoral ativa (AU).

Immune surveillance, mainly mediated by CD8 + T lymphocytes, recognize and destroy malignant or altered cells. However, through immunosuppressive strategies, such as the signaling pathways of the programmed cell death ligand-1 (PD-L1) and human leukocyte antigen-G (HLA-G), these mutated cells often escape the antitumor immune response. The aim of this study was to investigate and compare the immunoexpression of PD-L1, HLA-G, CD8 and granzyme B (GrB) in the microenvironment of lip squamous cell carcinomas (LSCCs; n = 40), actinic cheilitis (ACs; n = 55), and healthy lip mucosa (HLM; n = 10). The samples were submitted to immunohistochemistry and the analysis followed a semi-quantitative (PD-L1 and HLA-G) and quantitative methods (CD8 and GrB). Protein expression was compared between the three groups of samples, as well as with the lesion's clinicopathologic parameters and overall survival of patients with LSCC. Correlation between proteins and the type of tumor microenvironment according to a presence of PD-L1 and CD8 were also evaluated. Statistical tests included Fisher's exact, Mann-Whitney, Kruskal-Wallis, Spearman's correlation, as well as the log-rank for comparison of the overall survival built through Kaplan-Meier method. Significance was set at p < 0.05. The CD8+ and GrB+ cell numbers progressively increased from HLMs to LSCCs, with ACs exhibiting intermediate numbers (p < 0.01). Lower expression of these proteins was associated with lymph node metastasis and poor tumor differentiation (p < 0.05). PD-L1 and HLA-G expression in neoplastic cells/keratinocytes and stroma/connective tissue was significantly higher in LSCCs and ACs, compared to HLMs (p < 0.05). PD-L1 was not significantly associated with clinicopathological aspects of the lesions. Most LSCCs showed coexistence of PD-L1+ and CD8+ cells (72.5%) in the tumor microenvironment. PDL1 was directly correlated to CD8+ and GrB+ lymphocytic infiltration in LSCCs (p < 0.05). Proteins expression was not associated with overall survival of LSCCs patients (p > 0.05). Our findings suggest that immunosuppressive molecules PD-L1 and HLA-G are consistently expressed from ACs and are maintained until advanced stages of LSCCs. The correlation between PD-L1 expression and the expression of CD8 and GrB in carcinomas suggests that that PD-L1 may appear as an escape mechanism against an active antitumor response (AU).

Evaluation of the commercial kit SIRE nitratase for detecting resistant mycobacterium tuberculosis in Brazil

Abstract INTRODUCTION: This study aimed to evaluate a new commercial kit, Kit SIRE Nitratase-PlastLabor, for testing the drug susceptibility of clinical Mycobacterium tuberculosis isolates. METHODS: The accuracy of the Kit SIRE Nitratase was evaluated by examining the susceptibility (streptomycin, isoniazid, rifampicin, and ethambutol) of 40 M. tuberculosis isolates, using the proportion method with Lowenstein-Jensen medium or the BACTEC MGIT 960 system. RESULTS: The detection accuracy for streptomycin, isoniazid, rifampicin, and ethambutol was 95%, 97.5%, 100%, and 80%, respectively. CONCLUSIONS: The exceptional accuracy demonstrated by Kit SIRE Nitratase for isoniazid and rifampicin makes the kit an attractive option for screening M. tuberculosis strain resistance.

Calcifying cystic odontogenic tumour associated with odontoma involving mixed dentition / Tumor odontogênico cístico calcificante associado a odontoma envolvendo dentição mista

Calcifying cystic odontogenic tumor (CCOT) is a benign tumor relatively uncommon that may be associated with other odontogenic lesions. This manuscript describes the case of a 9-year-old girl affected by CCOT associated with odontoma, presenting as a slight swelling along with prolonged retention of a maxillary primary central incisor. Radiographic examination showed a unilocular radiolucent lesion containing radiopaque material associated with an unerupted tooth. Taking into consideration the clinical diagnostic of odontoma, an excisional biopsy was carried out. Microscopic examination established the diagnosis of a CCOT associated with odontoma. The association of these lesions is rare in individuals in the first decade of life, particularly involving mixed dentition, as outlined in this case. The patient remains under care with no clinical signs of recurrence.

O tumor odontogênico cístico calcificante (TOCC) é um tumor odontogênico cístico benigno relativamente raro e pode estar associado a outras lesões odontogênicas. Este manuscrito descreve um caso de CCOT associado a odontoma em uma menina de 9 anos de idade, que se apresentou com um ligeiro aumento de volume associado a retenção prolongada de um incisivo central superior. O exame radiográfico revelou uma lesão radiolúcida unilocular contendo material radiopaco, associado a um dente incluso. Tomando em consideração o diagnóstico clínico de odontoma, uma biópsia foi realizada. O exame microscópico estabeleceu o diagnóstico de um TOCC associado a odontoma. A associação destas lesões é rara em indivíduos na primeira década de vida, particularmente envolvendo dentição mista, conforme descrito no presente caso. O paciente permanece sob cuidados sem sinais clínicos de recorrência.

Clinicopathological profile and management of 161 cases of actinic cheilitis

AbstractBACKGROUND:Actinic cheilitis (AC) is a potentially malignant disorder of the lip caused by chronic exposure to ultraviolet radiation from the sun.OBJECTIVES:To evaluate the clinical, demographic, morphological and therapeutic management in AC cases data associating to the histopathological grading.METHODS:Demographic, clinical and management data of 161 patients with AC were analyzed. In biopsied cases, two calibrated examiners performed histopathological grading by binary system.RESULTS:There was a prevalence of males (79.5%), aged 40 years or older (77.5%), light-skinned (85.7%), experiencing occupational exposure to sunlight (80.3%), with AC presenting clinically as white lesions (33.6%). Conservative treatment was adopted in 78 cases and biopsy in 83 cases (60.2% graded as low-risk AC). There were no significant associations between histopathological grading and gender (p= 0.509), age (p=0.416), ethnicity (p=0.388), occupational exposure to sunlight (p=1.000) or clinical presentation (p=0.803).CONCLUSION:This study reinforces the hypothesis that demographic and clinical characteristics of AC are not related to histopathological grading. Advice on protection from sun exposure should be encouraged to avoid progression of AC and invasive therapies.

Rapid tests for the detection of the Mycobacterium abscessus subsp. bolletii strain responsible for an epidemic of surgical-site infections in Brazil

A single strain of Mycobacterium abscessus subsp. bolletii, characterised by a particular rpoB sequevar and two highly related pulsed field gel electrophoresis patterns has been responsible for a nationwide outbreak of surgical infections in Brazil since 2004. In this study, we developed molecular tests based on polymerase chain reaction restriction-enzyme analysis (PRA) and sequencing for the rapid identification of this strain. Sequences of 15 DNA regions conserved in mycobacteria were retrieved from GenBank or sequenced and analysed in silico. Single nucleotide polymorphisms specific to the epidemic strain and located in enzyme recognition sites were detected in rpoB, the 3' region of the 16S rDNA and gyrB. The three tests that were developed, i.e., PRA-rpoB, PRA-16S and gyrB sequence analysis, showed 100%, 100% and 92.31% sensitivity and 93.06%, 90.28% and 100% specificity, respectively, for the discrimination of the surgical strain from other M. abscessus subsp. bolletii isolates, including 116 isolates from 95 patients, one environmental isolate and two type strains. The results of the three tests were stable, as shown by results obtained for different isolates from the same patient. In conclusion, due to the clinical and epidemiological importance of this strain, these tests could be implemented in reference laboratories for the rapid preliminary diagnosis and epidemiological surveillance of this epidemic strain.

Variabilidade genética de amostras de Salmonella Typhi isoladas de surto e de casos esporádicos ocorridos em Belém, Brasil / Genetic variability of Salmonella Typhi samples isolated from outbreaks and sporadic cases of typhoid fever in Belém, Brazil

INTRODUÇÃO: Salmonella Typhi é o agente da febre tifóide (doença caracterizada por febre, cefaléia, mialgia, artralgia, diarréia ou constipação), cujo quadro pode se complicar e levar o paciente a óbito. No Brasil, a febre tifóide é endêmica nas regiões Norte e Nordeste, com surtos ocorridos nos meses de intenso calor. OBJETIVO: Analisar e comparar a variabilidade genética de S. Typhi isoladas de surto e casos esporádicos de febre tifóide ocorridos em determinado período na cidade de Belém (PA). MATERIAL E MÉTODOS: Foram analisadas 20 amostras de S. Typhi: 10 isoladas de um surto ocorrido no bairro do Guamá, Belém, entre os meses de dezembro/2005 e março/2006, e 10 de casos esporádicos ocorridos em diferentes localidades da mesma cidade e no mesmo período do surto. A caracterização genética foi realizada pela análise do perfil de macrorrestrição obtido pela enzima XbaI e definido por eletroforese em gel de campo pulsado (PFGE). RESULTADOS: A análise de XbaI-PFGE das amostras estudadas demonstrou uma similaridade genética de 83 por cento a 100 por cento. CONCLUSÃO: Este estudo pôde demonstrar a relação clonal das amostras S. Typhi causadoras de surto e de casos esporádicos de febre tifóide ocorridos na cidade de Belém no período de dezembro/2005 a março/2006.

BACKGROUND: Salmonella Typhi is the causative agent of typhoid fever, illness characterized by fever, migraine, myalgia, arthralgia, diarrhea or constipation, which may have complications and cause death. In Brazil, the typhoid fever is endemic in the Northern and Northeastern regions, with outbreaks occurring in scorching months. OBJECTIVE: To analyse and compare the genetic variability of S. Typhi strains isolated from outbreaks and sporadic cases of typhoid fever occurred in the city of Belém (PA) between December 2005 and March 2006. MATERIAL AND METHODS: Twenty samples of S. Typhi were analyzed: 10 of them were isolated from an outbreak occurred in Guamá neighborhood in Belém, between December 2005 and March 2006, and the other 10 were isolated from sporadic cases in different neighborhoods of the same city in the same outbreak period. The genetic characterization was performed by macrorestriction analysis of genomic DNA with XbaI enzyme defined by pulsed-field gel electrophoresis (PFGE). RESULTS: The Xbal-PFGE analysis of the studied samples revealed a genetic similarity of 83 percent to 100 percent. CONCLUSION: This study demonstrated the clonal relation between the S. Typhi samples from the outbreak and from the sporadic cases of typhoid fever occurred in the city of Belém between December 2005 and March 2006.

Avaliação de diferentes procedimentos para extração de DNA a partir de esfregaços corados pelo método de Ziehl-Neelsen / Evaluation of different methods of DNA extraction from baciloscopic tests of Ziehl-Neelsen stainig

O Pará é o estado com maior taxa de incidência de tuberculose (TB) na região Norte, com 36,3 casos por 100 mil habitantes em 2004. A obtenção de DNA de qualidade a partir de material fixado em lâminas coradas pelo método de Ziehl-Neelsen (ZN) supriria a falta de amostras representativas para estudos de genotipagem, contribuindo para melhor entendimento da dinâmica de transmissão da TB. O presente estudo avaliou quatro diferentes procedimentos para obtenção de DNA de M. tuberculosis a partir de 196 esfregaços corados pelo método de ZN, que utilizaram xilol e N-acetil-L-cisteína (NALC) em conjunto, de forma isolada ou nenhuma destas substâncias, e ainda dois protocolos de amplificação distintos, um com o adjuvante de PCR glicerol e outro com betaína. O percentual de amplificação foi superior para as amostras que foram submetidas ao procedimento que utilizou somente o xilol e ao que não utilizou nenhuma substância, sendo 77 por cento e 67 por cento, respectivamente, para o protocolo com glicerol, e de 98 por cento e de 96 por cento para o protocolo com betaína. A partir dos resultados obtidos, não é recomendado o uso de xilol ou NALC nos procedimentos de remoção das amostras e sugere-se a inclusão do adjuvante betaína nas reações, assim como o uso do DNA extraído de lâminas coradas pelo método de ZN em análises baseadas em PCR.

Nested-PCR do gene que codifica o antígeno b aplicada ao diagnóstico da tuberculose pulmonar / Nested-PCR for gene that encodes the antigen b applied to the diagnosis of pulmonary tuberculosis

A reação em cadeia da polimerase usada para amplificação de uma seqüência interna de um fragmento previamente amplificado (nested-PCR) foi investigada como uma alternativa complementar a pesquisa de bacilos álcool ácido resistentes e a cultura do Mycobacterium tuberculosis em meio de Lowenstein-Jensen. Foram investigadas 144 amostras de escarro de pacientes suspeitos de tuberculose encaminhados ao Laboratório de Tuberculose do Instituto Evandro Chagas em Belém, no período de junho de 2002 a dezembro de 2003. Das 144 amostras, 121 foram caracterizadas como tuberculose, 119 foram positivas na cultura, 95 na baciloscopia e 128 na nested-PCR. A sensibilidade da nested-PCR foi 96 por cento (116/121), enquanto a especificidade foi 48 por cento (11/23). A nested-PCR poderá ser uma ferramenta complementar para o diagnóstico da tuberculose, pois apresenta sensibilidade equivalente à cultura, no entanto, necessita de maiores avaliações visando minimizar o número de resultados falso-positivos.

The polymerase chain reaction used for amplifying an internal sequence of a previously amplified fragment (nested-PCR) was investigated as a complementary alternative for searching for alcohol-acid resistant bacilli and Mycobacterium tuberculosis cultures in Lowenstein-Jensen medium. 144 sputum samples were investigated from patients with suspected tuberculosis that were sent to the Tuberculosis Laboratory of the Evandro Chagas Institute in Belém, between June 2002 and December 2003. From the 144 samples, 121 were characterized as tuberculosis: 119 were positive in cultures, 95 under bacilloscopy and 128 using nested-PCR. The sensibility of the nested-PCR was 96 percent (116/121), while the specificity was 48 percent (11/23). Nested-PCR may be a complementary tool for diagnosing tuberculosis, since it presents sensitivity equivalent to that of cultures. However, further evaluations are needed with the aim of minimizing the number of false-positive results.

Diagnóstico molecular da tuberculose pulmonar / Molecular diagnosis of pulmonary tuberculosis

A reação em cadeia da polimerase (PCR) e suas variações, como a nested-PCR, têm sido destacadas como técnicas moleculares promissoras para o diagnóstico rápido da tuberculose (TB). No presente estudo avaliou-se a nested-PCR utilizando-se como marcadores moleculares a seqüência IS6110 e o antígeno b aplicados ao diagnóstico da TB. Foram submetidas a baciloscopia, cultura e nested-PCR 136 amostras clínicas de pacientes com suspeita de TB. O diagnóstico de tuberculose pulmonar foi atribuído a 116 pacientes e, desses, 97 foram multibacilares e 111 apresentaram cultura positiva para M. tuberculosis. As reações de nested-PCR identificaram 70 por cento (antígeno b) e 94 por cento (IS6110) dos casos paucibacilares. Os valores de sensibilidade determinados para cultura, nested-PCR do IS6110 e antígeno b foram 95 por cento, 98 por cento e 86 por cento, respectivamente. A especificidade foi de 100 por cento, 15 por cento e 45 por cento para cultura, nested-PCR do IS6110 e antígeno b, respectivamente. O diagnóstico molecular da tuberculose deve estar fundamentado na análise conjunta de vários parâmetros, como baciloscopia, cultura, manifestações clínicas, prova terapêutica e história prévia de tuberculose.

Chain reaction of polimerase (PCR) and its variations such as the nested-PCR have been outstanding as promising molecular techniques for the fast diagnosis of the tuberculosis (TB). In the present study the nested-PCR was evaluated using as molecular markers the sequence IS6110 and the antigen b applied to the diagnosis of TB. One hundred and thirty six clinical samples of patients with suspicion of TB were submitted to the baciloscopia, culture and nested-PCR. The diagnosis of pulmonary tuberculosis was attributed to 116 patients, of these, 97 were multi-bacilli and 111 presented positive culture for M. tuberculosis. The nested-PCR reactions identified 70 percent (antigen b) and 94 percent (IS6110) of the acid-fast smear negative cases of tuberculosis. The sensibility values determined for culture, and nested-PCR of IS6110 and antigen b were 95 percent, 98 percent and 86 percent, respectively. The specificity was 100 percent, 15 percent and 45 percent for culture, nested-PCR of IS6110 and antigen b, respectively. The molecular diagnosis of the tuberculosis should be based in the combined analysis of several parameters, as the baciloscopia, culture, clinical manifestations, therapeutic proves and previous history of tuberculosis.