Anaplasma phagocytophilum direct detection and exposure evidence in equines from two breeding farms from Minas Gerais State, Brazil / Detecção direta e evidência da exposição a Anaplasma phagocytophilum em equinos de dois haras em Minas Gerais, Brasil

Equine granulocytic anaplasmosis is caused by Anaplasma phagocytophilum, a gram negative, obligatory intracellular bacterium, member of Anaplasmataceae family, included in the Rickettsiales order. Little is known about the disease, transmission dynamics, genetic diversity and prevalence in Minas Gerais state, Brazil. This work aimed to do a serosurvey using indirect immunofluorescent assay (IFA) test and evaluation of buffy coat smears, and nested polymerase chain reaction (PCR) as diagnostic methods, to determine the disease situation in horses from two manga-larga marchador breeding farms located in the municipalities of Ataléia e São Vicente de Minas, in Minas Gerais state. It was found that 76% (131/172) of the animals were considered reactive for IFA test, and the total of 12.8% was positive at buffy coat smears analysis. At PCR analysis, it was found 1.94% of the samples positive to the infection. Those samples were sequenced and showed 96% of similarity to A. phagocytophilum from a Ixodes ricinus tick. There is a high frequency of animals with the evidence of contact to A. phagocytophilum on the two evaluated properties in this study, which was proved by positiveness in PCR analysis. New researches must be carried out to better understand the epidemiologic and clinical dynamic of the disease in the state of Minas Gerais.(AU)

A anaplasmose granulocítica equina é causada por uma bactéria gram-negativa, intracelular obrigatória, membro da família Anaplasmataceae, incluída na ordem Rickettsiales e denominada de Anaplasma phagocytophilum. Pouco se sabe sobre a doença, sua dinâmica de transmissão, diversidade genética e prevalência em Minas Gerais, Brasil. Este trabalho teve por objetivo realizar o levantamento sorológico utilizando a reação de imunofluorescência indireta, avaliação direta de capa leucocitária e nested reação em cadeia da polimerase (PCR) como métodos diagnósticos, a fim de avaliar a situação da doença em dois haras de criação de cavalos manga-larga marchador localizados nas cidades de Ataléia e São Vicente de Minas, no estado de Minas Gerais. Foi encontrada prevalência de 76% (131/172) de animais reativos para a reação de imunofluorescência indireta, quando todos os animais das duas propriedades e das duas coletas foram agrupados, e 12,8% dos animais foram positivos na avaliação da capa leucocitária. A reação de imunofluorescência indireta detectou 1,94% das amostras como positivas para o agente. Essas amostras foram submetidas ao sequenciamento de nucleotídeos, e foi observada similaridade de 96% com A. phagocytophilum proveniente de carrapatos Ixodes ricinus. Existe alta prevalência de animais positivos para a infecção por A. phagocytophilum, o que foi provado pela positividade dos animais à PCR. Novas pesquisas devem ser conduzidas a fim de entender a dinâmica epidemiológica e clínica da doença no estado de Minas Gerais.(AU)

Could mesenchymal stem cell therapy help in the treatment of muscle damage caused by Bothrops alternatus venom? / Podem as células tronco mesenquimais auxiliarem no tratamento da lesão muscular causada pelo veneno de Bothrops alternatus?

ABSTRACT: The aim of this study was to evaluate the use of mesenchymal stem cells (MSC) in the treatment of myonecrosis induced by Bothrops alternatus venom in rats. Seventy-five male adult Wistar rats were divided into three experimental groups. G1 and G2 were injected in the gastrocnemius muscle with 120μg of B. alternatus venom, while G3 received 200μL of PBS only. Three days after the venom injection, 12 rats from G1 were treated with 5.0 x 106 MSC in PBS, whereas G2 and G3 rats received PBS. Every three days, blood and muscle samples of five animals from each group were taken for serum biochemical and pathological analyses. Histological examinations showed more intense muscle lesions following MSC treatment, characterized by disorganization and loss of muscle fibers, with focal necrosis and inflammatory infiltration by mononuclear cells. In conclusion, the use of MSC for the treatment of local damage caused by inoculation of B. alternatus venom impaired muscle regeneration and interfered in the healing process.

RESUMO: O objetivo do presente trabalho foi avaliar a utilização das células tronco mesenquimais (MSC) no tratamento da mionecrose induzida pelo veneno de Bothrops alternatus em ratos. 75 ratos Wistar adultos foram distribuídos em três grupos experimentais. G1 e G2 receberam 120μg de veneno de B. alternatus, enquanto o G3 recebeu apenas 200μL de PBS. Três dias após a administração do veneno, os ratos do grupo G1 foram tratados com 5.0 x 106 MSC, enquanto G2 e G3 receberam exclusivamente PBS. A cada três dias, amostras de sangue e tecido muscular foram coletadas de cinco animais de cada grupo para avaliação bioquímica sérica e patológica, respectivamente. A análise histológica revelou lesão muscular mais intensa após a aplicação das MSC, caracterizada pela desorganização e perdas das fibras musculares, com necrose focal e infiltrado inflamatório mononuclear. É possível concluir que a utilização das MSC para o tratamento da lesão local causada pela inoculação do veneno B. alternatus deteriorou a regeneração muscular e interferiu com o processo de cicatrização.

Periodontopathogens, Candida spp. and immunological aspects in type 2 diabetes mellitus patients with chronic periodontitis

This study aimed to evaluate clinical, microbiological and immunological parameters in type 2 diabetes mellitus (DM) in comparison with normoglycemic patients (NDM). Glycemic and lipid profiles and periodontal clinical status were determined for thirty-three patients (17 DM and 16 NDM). The presence of periodontopathogens and species of Candida in subgingival sites were determined by polymerase chain reaction and immunological parameters by ELISA assays. All glycemic and clinical parameters evaluated were higher in the DM group, with statistical difference for fasting glucose, glycated-hemoglobin, and periodontal parameters. Lipid profile (except triglycerides), levels of TNF-α and myeloperoxidase and the prevalence of the tested microorganisms were similar between the groups, except for Candida albicans and Candida glabrata, which was higher in the DM group. In conclusion, although microbiological and immunological parameters were similar in the DM and NDM groups, periodontitis and the levels of some species of Candida were more severe in DM patients. (AU)

Validación del cuestionario para graduación de dolor crónicoen pacientes chilenos con patología musculoesquelética / Validation of the graduation of chronic pain in patients with chilean musculoskeletal pathology

Introducción: el dolor crónico es una condición que afecta a 1 de cada 5 personas en el mundo, comprometiendo diferentes áreas de la calidad de vida. El cuestionario para graduación de dolor crónico (CGDC) fue desarrollado como una forma de evaluar y monitorear a estos pacientes, con altos niveles de fiabilidad y validez. Objetivos: Desarrollar una versión española del CGDC, adaptado culturalmente a Chile y determinar su validez y fiabilidad, en el Hospital Clínico de la Universidad de Chile. Material y métodos: El cuestionario fue traducido y adaptado culturalmente de acuerdo a las recomendaciones internacionales. Se aplicó SF-36 v2.0 en 130 pacientes condolor musculoesquelético crónico (más de 6 meses). La fiabilidad se calculó con Alfa de Cronbach y el índice de validez se evaluó mediante la comparación de las respuestas de la CGDC para cada categoría con las subescalas del SF-36 v.2. Resultados: La versión chilena de la CGDC fue válida. Se obtuvieron altos niveles de confiabilidad con alfa de Cronbach> 0,7. Se observaron correlaciones significativas del SF -36,especialmente con las subescalas que tienen alta capacidad de medir el dolor y la salud física (p <0,01). Conclusiones: Los resultados presentados aquí confirman la fiabilidad y validez de la versión chilena del CGDC en la evaluación de pacientes con dolor musculoesquelético crónico.

Introduction: chronic pain is a condition that affectss 1 in every 5 people in the world, compromising different areas of quality of life. The Chronic Pain Graded questionnaire (CPG) was developed as a form to assess and monitor these patients, with high levels of reliability and validity. Objectives: To develop a spanish version of the chronic pain graded questionnaire, culturally adapted to Chile determine its reliability and convergent construct validity , in the University of Chile Clinical Hospital. Materials and Methods: The chronic pain graded questionnaire was translated and culturally adapted accordingt o international recommendations. It was applied with SF -36 v2.0 questionnaire in 130 patients with chronic musculoskeletal pain (more than 6 months). The reliability was calculated with de Alpha the Cronbach Index and de validity was assessed by comparing the responses of the CPG for each category with the subscales of SF-36 v.2. Results: The Chilean version of the CBDC was valid. High levels of reliability was obtained with Cronbach’s alpha > 0.7. Compared with the SF -36 significant correlations were observed, especially with the SF -36 subscales having high ability to measure pain and physical health ( P < 0.01). Conclusions: The results presented here confirm the reliability and validity of the Chilean version of the chronic pain graded questionnaire in the evaluation of patients with chronic musculoskeletal pain.

Morfologia de potros da raça Brasileiro de Hipismo / Morphology of Brasileiro de Hipismo foals

Objetivou-se avaliar a morfologia de potros da raça Brasileiro de Hipismo, utilizando análise de imagens digitais e medidas de perímetros corporais para estimativa de índices corporais dos animais. As mensurações lineares e angulares foram obtidas utilizando imagens digitais dos potros com câmera acionada pelo aplicativo Simi Reality Motion Systems®. As medidas foram baseadas em pontos anatômicos realçados com 19 marcadores reflexivos. A análise estatística descritiva dos resultados foi realizada utilizando o programa SAS. Os potros estavam em crescimento, com aumento das medidas lineares de altura na cernelha e na garupa, comprimento do corpo, distância escápulo-metacarpofalângica e nos comprimentos do pescoço, da perna e do antebraço. Os ângulos úmero-radial, metacarpo-falângico, fêmoro-tibial, tíbio-metatarsico aumentaram com a idade, enquanto que os ângulos da cabeça, do pescoço, coxo-femoral, metatarso-falângico, escápulo-solo e coxo-solo foram variáveis entre os momentos de avaliação. Os potros foram classificados em mediolíneos e hipermétricos pelos índices Corporal e Dáctilo-torácico, respectivamente. A avaliação do crescimento dos potros foi possível através da imagem digital.

This research aimed to evaluate the morphology of Brasileiro de Hipismo foals using digital image analysis and girth measurements to estimate body indexes of animals. The linear and angular measurements were obtained by digital images of the foals with camera triggered by Simi Reality Motion Systems®. The measures were based on anatomical landmarks highlighted with 19 reflective markers. A descriptive statistical analysis of the results was performed using the SAS program. The foals were growing, with increased linear measurements of height at withers and croup, body length, distance from shoulder-fetlock and neck length, leg length and forearm length. The angles humerus-radial, metacarpal-falangeano, femoro-tibial, tibio-metatarsal increased with age, whiles the angles of the head, neck, coxofemoral, metatarsal-falangeano, shoulder-floor and coxae-floor were variable among the periods. The foals were classified as medium shape and hypermetric by Corporal Index, Dactilo-thoracic Index, respectively. Assessment of growth of the foals was possible through a digital image.

Antinociceptive effects of an extract, fraction and an isolated compound of the stem bark of Maytenus rigida

The antinociceptive activity of the Maytenus rigida Mart. (Celastraceae) ethanol extract and its ethyl acetate fraction as well as of (-)-4'-methylepigallocatechin (1), a previously isolated compound, was demonstrated in vivo. ED50 for 1 in the writhing test was 14.14 mg/kg. The acetic acid-induced writhing was inhibited by 98.4, 84.4, and 58.3%, respectively, when mice were treated with the ethanol extract, ethyl acetate fraction, and 1. In the hot plate test, mice pretreated with 1 showed significantly increased reaction times (60-89%). Oral administration of 1 significantly inhibited first and second phases of the formalin-induced pain (50 and 26.5%, respectively), whereas indomethacin inhibited only the second phase of the test (41.2%). Ethanol extract and its fraction showed effects on inflammatory pain, while neurogenic and inflammatory pain suppression by 1 is a strong indication of the presence of both central and peripheral effects and suggests its analgesic and anti-inflammatory potential.

Therapeutic approaches for tumor necrosis factor inhibition

Tumor necrosis factor (TNF) consists of an inflammatory cytokine essential for homeostasis and organism defense. Despite its physiological relevance, both increased biosynthesis and release of TNF lead to the exacerbation of inflammatory and oxidative responses, which are related to the pathogenesis of a host of diseases of an inflammatory, autoimmune and/or infectious nature. In this context, effective therapeutic approaches for the modulation of TNF have been the focus of research efforts. Approximately one million individuals worldwide have been treated with biotechnological inhibitors of this cytokine, the so-called anti-TNF biopharmaceuticals. However, given the high risk of infection and the limitations related to cost and administration routes, new therapeutic approaches aimed at biological targets that directly or indirectly modulate the production and/or activation of TNF appear promising alternatives for the discovery of new anti-inflammatory and immunomodulatory orally active drugs and are therefore discussed in this paper.

O fator de necrose tumoral (do inglês, tumor necrosis factor - TNF) consiste em uma citocina inflamatória essencial para a homeostase e defesa do organismo. A despeito de sua relevância fisiológica, o aumento da biossíntese e liberação do TNF conduzem à exacerbação das respostas inflamatória e oxidativa, as quais estão relacionadas à patogênese de várias doenças de natureza inflamatória, auto-imune e/ou infecciosa. A busca por abordagens terapêuticas eficientes na modulação do TNF tem sido alvo de diversos esforços de pesquisa. Aproximadamente um milhão de pessoas ao redor do mundo já foi tratado com inibidores biotecnológicos desta citocina, os chamados biofármacos anti-TNF. Entretanto, em face ao elevado risco de infecções e as limitações relacionadas ao custo e a via de administração, novas abordagens terapêuticas com foco em alvos que modulem, de forma direta ou indireta, a produção e/ou ativação do TNF surgem como alternativas promissoras para a descoberta de novos fármacos antiinflamatórios e imunomoduladores ativos por via oral e serão discutidas neste trabalho.

Experiencia del bloqueo paravertebral en sensibilización espinal segmentaria / Paraspinous block experience in spinal segmental sensitization

La Sensibilización Espinal Segmentaria (SES) corresponde a un estado hiperactivo de un segmento espinal en reacción a un foco irritativo, el cual bombardea constantemente el ganglio sensorial con estímulos. Esto se traduce en dolor espontáneo, hiperalgesia y adolinia. El Bloqueo Paraespinoso (BPE) ayudaría a la desensibilización gradual del segmento espinal comprometido. Objetivo: Mostrar manifestaciones de la SES y los efectos de Bloqueo Paraespinal en pacientes con dolor crónico de extremidad superior atendidos en el Servicio de Medicina Física y Rehabilitación del HCUCH entre abril - agosto 2008. Material y Métodos: Estudio descriptivo - retrospectivo de pacientes que consultan por dolor crónico de extremidad superior, que presentaron signos de SES en atención ambulatoria del Servicio de Medicina Física y Rehabilitación del HCUCH. Se evaluó características demográficas y clínicas, y respuestas tras BPE. Resultados: Once pacientes presentaron signos de SES. El 100 por ciento fue de sexo femenino, con una mediana edad de 44 años. El 63,6 por ciento presentó Epicondilitis como diagnóstico inicial, seguido de Lesión de MAnguito Rotator (36 por ciento) y el 36 por ciento presentó mas de una patología. El EVA inicial fue en promedio de 8,9. El territorio afectado más frecuente fue C6 - C7 (36 por ciento). Tras el BPE, el dolor en reposo disminuye a un EVA promedio de 2,8 (diminución de un 68,3 por ciento). Incidentalmente, se observó ganancia en los rangos articulares de los segmentos comprometidos. Discusión: La búsqueda y tratamiento de la SES mediante el BPE puede ser una herramienta útil en el manejo de pacientes con dolor crónico musculoesquelético.

Introduction: Spinal Segmental Sensitization (SSS) relates to an hyperactive state of one spinal segment reacting to a source of irritation that is constantly stimulating the dorsal ganglion. This translates into spontaneous pain, hyperalgesia and allodinia. The Paraspinus Block (PB) technique would prove useful to desensitize the reacting segment. Objetive: To show SSS manifestations and the effects of Paraspinus Block in patients with chronic pain in upper limbs treated at Hospital Clinico Universidad de Chile Physical and Rehab Med Center from April through August 2008. Materials and Method: Descriptive and retrospective study of Physical and Rehab Med Center outpatients with upper limb chronic pain showing signs of SSS. Demographic and clinical information was evaluated as well as response to PB treatment. Results: Eleven patients showed signs of SSS of which 100 percent were female in their mid forties. 63.6 percent were initially diagnosed with Epycondilitis while 36 percent with Rotator Cuff Tear and 36 percent with more than one disease. The initial average VAS was 8.9. The most affected area was C6 - C7 (36 percent). After PBE pain while resting is reduced to an average VAS of 2.8 (a 68.3 percent reduction). We also observed a gain in irritated segments. Discussion: The search and treatment of SSS using PBE could be a useful tool for pain management in patients with musculoskeletal chronic pain.

Estratégias em química medicinal para o planejamento de fármacos / Strategies in medicinal chemistry to the discovery of new lead-compounds

Este artigo ilustra a aplicação das estratégias de hibridação molecular e bioisosterismo no planejamento estrutural de novos candidatos a protótipos de fármacos com propriedades analgésicas, antiiinflamatórias e antitrombóticas, com base no mecanismo de ação pretendido, explorando, inclusive, produto natural brasileiro abundante como padrão molecular básico...