α, ß-Amyrin prevents steatosis and insulin resistance in a high-fat diet-induced mouse model of NAFLD via the AMPK-mTORC1-SREBP1 signaling mechanism

Nonalcoholic fatty liver disease (NAFLD), characterized by hepatosteatosis and steatohepatitis, is intrinsically related to obesity. Our previous study reported on the anti-obese activity of α,β-amyrin (AMY), a pentacyclic triterpene isolated from Protium heptaphyllum. This study investigated its ability to prevent fatty liver and the underlying mechanism using the mouse model of NAFLD. NAFLD was induced in male Swiss mice fed a high fat diet (HFD) for 15 weeks. The controls were fed a normal chow diet (ND). The mice were simultaneously treated with AMY at 10 and 20 mg/kg or fenofibrate at 50 mg/kg. Lipid levels along with metabolic and inflammatory parameters were assessed in liver and serum. The liver sections were histologically examined using H&E staining. RT-qPCR and western blotting assays were performed to analyze signaling mechanisms. Mice fed HFD developed severe hepatic steatosis with elevated triglycerides and lipid droplets compared with ND controls. This was associated with a decrease in AMP-activated protein kinase (AMPK) activity, an increase of mechanistic target of rapamycin complex 1 (mTORC1) signaling, and enhanced sterol regulatory element binding protein 1 (SREBP1) expression, which have roles in lipogenesis, inhibition of lipolysis, and inflammatory response. AMY treatment reversed these signaling activities and decreased the severity of hepatic steatosis and inflammatory response, evidenced by serum and liver parameters as well as histological findings. AMY-induced reduction in hepatic steatosis seemed to involve AMPK-mTORC1-SREBP1 signaling pathways, which supported its beneficial role in the prevention and treatment of NAFLD.

MDGC-MS analysis of essential oils from Protium heptaphyllum (Aubl. ) and their antifungal activity against Candida specie / Análise MDGC-MS de óleos essenciais de Protium heptaphyllum (Aubl. ) e sua atividade antifúngica contra espécies de Candida

ABSTRACT Protium heptaphyllum is found in the Amazon region, and in various Brazilian states and South American countries. Also Known as almecega, it produces an oil resin used in traditional medicine as analgesic, anti-inflammatory, cicatrizant and expectorant, it is rich in pentacyclic triterpenes and essential oil. The main objective of this study was to analyze the chemical composition of P. heptaphyllumresin (OEPh) over different extraction times and to evaluate their antifungal activity against Candida species, obtained from gardeners with onychomycosis, using the disk diffusion method. The OEPh was obtained by hydrodistillation and analyzed by Multidimensional Gas Chromatography coupled with Mass Spectrometry (MDGC / MS). Candida species were obtained from lesions on the nails of horticulturist from a community garden in the city of Teresina, Piauí, Brazil. The antifungal activity in concentrations of 1000 µg/L, 500 µg/L and 250 µg/L, PROTOCOL M44-A2 (CLSI 2009) OEPh was tested. The main constituents identified were: l-limonene, α-terpineol, p-cineol, o-cymene and α-phellandrene, however, its composition varies significantly with extraction time. All species, except C. rugosa, were inhibited with halo (≥ 14 mm) at 1000 μg / L. C. krusei is naturally resistant to the drug fluconazole, but when tested with OEPh the clinical species (case 9) demonstrated sensitivity in three dilutions (halo ≤ 10 ≥ 14) and the standard strain was inhibited at concentration of 1000 μg/Lg / L (halo 14mm). A similar situation also occurred with the standard strain of C. parapsilosis (halo ≥ 11mm). OEPh has considerable antifungal activity, which merits further investigation for alternative clinical applications, since this species is widely distributed in our community, and it presents good yields, and also has important therapeutic applications.

RESUMO Protium heptaphyllum é encontrada na região amazônica, em vários estados do Brasil e países da América do Sul. Conhecida como almecega produz uma resina oleosa usada na medicina popular como analgésica, antiinflamatória, cicatrizante e expectorante, é rica em triterpenos pentaciclicos e óleo essencial. O objetivo principal do presente trabalho foi analisar a composição química do óleo essencial da resina P. heptaphyllum (OEPh) em diferentes tempo de extração e avaliarsuaatividade antifúngica contra espécies de Candida, isoladas de horticultores com onicomicoses, por método de disco-difusão. O OEPh foi obtido por hidrodestilação, analisado por Cromatografia Gasosa Multidimensinal Acoplada a Espectrometria de Massas (MDGC/MS). As espécies de Candida foram obtidas de lesões nas unhas de horticultores de uma horta comunitária na cidade de Teresina, Piauí, Brasil. Testou-se a atividade antifúngica do OEPhnas concentrações de 1000 μg/L, 500 μg/L e 250 μg/L, protocolo M44-A2 (CLSI 2009). Os principais constituintes identificados foram l- limoneno, α-terpineol, p-cineol, o-cimeno e α-felandreno, entretanto, sua composição varia significativamente em decorrência do tempo de extração. Todas as espécies, exceto a C. rugosa, foram inibidas com halo ( Χ ≥ 14 mm) na concentração de 1000 μg/L. C. krusei é naturalmente resistente ao fármaco fluconazol, mas quando testado com OEPh,a espécie clínico (caso 9) demonstrou sensibilidade nas três diluições (halo Χ ≤ 10 ≥ 14) e a cepa padrão foi inibida na concentração de 1000 μg/L (halo Χ 14mm). Fato semelhante também ocorreu com a cepa padrão de C. parapsilosis (halo Χ ≥ 11mm). O OEPh possui atividade antifúngica considerável, merecendo uma investigação mais aprofundada para aplicações clínicas alternativas, uma vez que esta espécie é amplamente distribuída em nossa comunidade, apresenta bom rendimento e, ainda, aplicações terapêuticas importantes.

Efeito da fração não-oleosa doRicinus communisna biodistribuição comTecnécio-99m em camundongos / Effect of non-oily fraction ofRicinus communis on the bio distribution with Technetium-99m in mice

RESUMO Ricinus communisé uma planta pertence à família Euforbiaceae. A partir de seus frutos é extraído o óleo de rícino e outra fração altamente tóxica (fração não-oleosa). Neste estudo, foi avaliado o efeito da fração não oleosa de R. communis(FNORC) na biodistribuição do Tc-99m em camundongos sadios e em animais transplantados com Sarcoma-180 (S180). Para avaliação da biodistribuição do Tc-99m, o percentual de atividade radioativa total injetada (ATI%) foi mensurada utilizando um contador gama. Os resultados mostraram que, em animais sadios, houve aumento da captação do Tc-99m após o tratamento com FNORC pelos rins, cérebro e estômago. Apenas houve diminuição da absorção deste radioisótopo nos músculos. Em animais com tumor, houve redução significativa da captação do Tc-99m no coração, pulmões e tumor, e foi observado o aumento da captação apenas nos rins. Foi observada também a diminuição da concentração das proteínas plasmáticas totais em animais com tumor após tratamento com FNORC e atividade antitumoral significativa. Diante desses dados, conclui-se que fração não oleosa de R. communis possui atividade antitumoral significativa, porém altera a biodistribuição do Tc-99m, fato este que pode resultar em falhas nos processos de diagnóstico por imagem que utilizem este radioisótopo como marcador.

ABSTRACT Ricinus communis is a plant belonging to the Euforbiaceae family. From its fruits, it is extracted the castor oil and another highly toxic fraction (non-oily fraction). In this study, we evaluated the effect of the non-oily fraction of R. communis (NOFRC) on the bio distribution of Tc-99m in healthy mice and in animals transplanted with Sarcoma-180 (S180). To evaluate the bio distribution of theTc-99m, the percentage of the total injected radioactivity (% TIR) was measured through a gamma counter. There was an increase in the capitation of Tc-99m after the treatment with NOFRC in the kidneys, brain, and stomach of healthy animals. A decrease in the reception of this radionuclide was only found in the muscles. In animals with tumour, there was a significant reduction in the uptake of Tc-99m in the heart, lungs and tumour. An increase in the capitation was only observed in the kidneys. It was also observed a reduction in the total plasma protein concentration in animals with tumours after the treatment with NOFRC, besides significant antitumor activity. We concluded that there is antitumor activity in the NOFRC, but that it alters the biodistribution of Tc-99m, a fact which may result in failures in the diagnostic imaging process using this radioisotope as a marker.

Plantas medicinais utilizadas pela população do município de Lagarto- SE, Brasil ­ ênfase em pacientes oncológicos / Medicinal Plants Used by the Population of Lagarto, SE, Brazil ­ Emphasis in Cancer Patients

RESUMO A utilização de plantas com fins medicinais é uma das mais antigas formas de prática medicinal da humanidade. Entretanto, o conceito de "natural"; contribui para o pensamento popular e errôneo de que Plantas Medicinais (PM) são sinônimos de produtos seguros, o qual pode ocasionar no desenvolvimento de efeitos adversos ou interações medicamentosas. Neste contexto, foi realizado levantamento etnofarmacológico das Plantas Medicinais (PM) utilizadas no município de Lagarto, SE, Brasil, com ênfase de seu uso por pacientes oncológicos. Um total de 706 moradores foram entrevistados. Foram citadas 80 plantas, das quais 57 foram identificadas em nosso laboratório. O uso de plantas medicinais (MP) para fins terapêuticos foi relatada por 336 (47,65%) entrevistados. As MPs mais utilizados foram: Erva-Cidreira (Lippiaalba (Mill) N. E. Brown - 103, 30,8%), Boldo (Plectranthus barbatus Andr. - 53, 15,7%), e Capim-Santo (Cymbopogon citratus (D.C.) Stapf - 49, 14,6%). Dos entrevistados que relataram o uso de MPs, metade (360, 50,95%) comunicaram ao médico, mas não receberam orientações específicas. As doenças crônicas mais comuns identificadas foram: hipertensão arterial (144, 20,34%), câncer (55, 7,81%) e diabetes (41, 5,89%). Dos pacientes com câncer, cerca de 40% (22) relataram utilizar PMs concomitante com quimioterapia, dado alarmante, visto a possibilidade de interações medicamentosas entre PMs e antineoplásicos. Diante desses dados, foi observado o uso de PM pela população de Lagarto, SE, e por pacientes oncológicos dessa região, porém, sem os mesmos receberem orientações de um profissional qualificado.

Medicinal Plants Used by the Population of Lagarto, SE, Brazil ­ Emphasis in Cancer Patients. The use of plants for medicinal purposes is one of the oldest forms of medical practice. However, the concept of "natural" contributes to the popular and erroneous thinking that Medicinal Plants (PM) are synonymous of safe products, which may result in the development of adverse effects or drug interactions, particularly in cancer patients. In this context, it was conducted an ethnopharmacological survey of MP used in Lagarto, SE, Brazil, with emphasis in cancer patients. A total of 706 residents were interviewed, and they mentioned 80 plants, from which 57 were identified. The use of medicinal plants (MP) for therapeutic purpose was reported by 336 (47.65%) of the participants. The most used MPs were as follows: Bushy Lippia (Lippia alba (Mill) N. E. Brown­ 103, 30.8%), Boldo (Plectranthus barbatus Andr. ­ 53, 15.7%), and Lemon Grass (Cymbopogon citratus (D.C.) Stapf. ­ 49, 14.6%). From the participants who reported the use of MPs, half of them (360, 50.95%) spoke with a doctor but did not receive specific guidelines. The most common chronic diseases identified were: arterial hypertension (144, 20.34%), cancer (55, 7.81%) and diabetes (41, 5.89%). From these cancer patients, approximately 40% of them (22) used MP concomitantly with chemotherapy, an alarming discovery, as drug interactions are possible between MPs and antineoplastics. Given these data, we observed the use of the PM by the population of Lagarto, SE, and by cancer patients of this region, but they did not receive guidance from a qualified professional

Erythropoietin reduces the expression of myostatin in mdx dystrophic mice

Erythropoietin (EPO) has been well characterized as a renal glycoprotein hormone regulating red blood cell production by inhibiting apoptosis of erythrocyte progenitors in hematopoietic tissues. EPO exerts regulatory effects in cardiac and skeletal muscles. Duchenne muscular dystrophy is a lethal degenerative disorder of skeletal and cardiac muscle. In this study, we tested the possible therapeutic beneficial effect of recombinant EPO (rhEPO) in dystrophic muscles in mdx mice. Total strength was measured using a force transducer coupled to a computer. Gene expression for myostatin, transforming growth factor-β1 (TGF-β1), and tumor necrosis factor-α (TNF-α) was determined by quantitative real time polymerase chain reaction. Myostatin expression was significantly decreased in quadriceps from mdx mice treated with rhEPO (rhEPO=0.60±0.11, control=1.07±0.11). On the other hand, rhEPO had no significant effect on the expression of TGF-β1 (rhEPO=0.95±0.14, control=1.05±0.16) and TNF-α (rhEPO=0.73±0.20, control=1.01±0.09). These results may help to clarify some of the direct actions of EPO on skeletal muscle.

Femoral biomechanic and microtomography from male ratssubmitted to dietary restriction supplemented with sucrose

Food restriction reduces body weight and influence bone mass and also is correlated with bone mineraldensity (BMD). Mechanisms have been proposed for the loss of BMD after body weight reduction, includingreduced energy intake. Growing 8 wk-old Wistar male rats were randomly divided into Control and Calorierestriction associated with sucrose 30% (CRS). These animals were subjected to intermittent food restrictionduring 8 weeks and had free access to tap water and sucrose30% in distilled water. The rats were euthanizedat the end of week 8, blood collected from abdominal aorta artery, femurs cleaned of adherent soft tissues,scanned using dual energy X-ray absorptiometry, structural and material properties determined by three-pointbending testing in the mid-diaphyseal region, bone surface tested in a microhardness tester and microstructurewas assessed in a microcomputer tomography. In CRS animals body weight decreased significantly relative tothe Control animals. There was a clear option for high-sucrose beverage in CRS animals. No difference wasobserved in biochemical, densitometric and biomechanical analyzes. Results from micro CT showed onlysignificant difference in connectivity of trabecular bone. It has been suggested that rats submitted to foodrestriction consumed sugar not because of its inherent palatability, but in order to alter their macronutrientbalance and animals need to meet energy demands in high-sucrose.

High-sucrose effect on bone structure, hardness and biomechanics in an obesity model using Wistar male rats

Introduction: Excessive consumption of sugar-sweetened beverage is positively related to overweight. Despite the epidemic of childhood obesity, body mass can have a positive or negative effect on bone health. Material and methods: Wistar rats 8 weeks olds were randomly assigned to consume water (Control group, n = 10), sucrose 30% (HS group, n = 10) and water + sucrose 30% (WHS group, n = 14) for 8 weeks. All animals received standard laboratory chow ad libitum. Femur measurements included microhardness, bone mineral density (BMD) by DXA, mechanical compression test and microcomputed tomography (microCT) analysis. Results: We observed significant difference in final body weight in HS and WHS groups, significant increase in triacylglycerol/fructosamine in HS and WHS groups, significantly high BMD in WHS group, increased periosteal/endosteal cortical microhardness in WHS group. Compared with control, microCT parameters evidenced lower amount of connected trabecular bone, decreased bone volume, lower trabecular number with high trabecular separation in distal epiphysis in WHS animals. Conclusion: High-sucrose consumption causes obesity induced by a liquid diet with negative effects on cancellous bone.

Caracterização epidemiológica da raiva bovina no estado de Mato Grosso, Brasil, no período de 1996 a 2006 / Epidemiological characterization of bovine rabies in the state of Mato Grosso, Brasil, in the period from 1996 to 2006

Um levantamento epidemiológico de todos os casos de raiva bovina ocorridos no Estado de Mato Grosso durante os anos de 1996 a 2006 foi realizado pela análise da curva epidêmica, por meio do cálculo do somatório, média, desvio padrão, valores máximos e mínimos da ocorrência de casos de raiva nos três ecossistemas; análise estatística entre o efetivo bovino nos três biomas do estado e o número de casos de raiva bovina; a distribuição sazonal e a situação da doença no estado em relação ao panorama nacional. Para organização dos dados, levou-se em consideração o mês e o ano de ocorrência da raiva e a região geográfica de origem do material. Os diferentes municípios envolvidos foram classificados conforme sua localização nas regiões de Cerrado, Pantanal e Amazônia. Constatou-se que a variação no número de casos positivos da enfermidade tem sido crescente nos últimos anos, sendo que o maior número de casos deu-se no bioma Cerrado, que apresentou uma tendência de aumento anual. A partir do ano de 2004, o ecossistema Cerrado passou da situação de enzootia para epizootia, ficando pois, evidente que o principal problema de raiva em bovinos no Estado de Mato Grosso está situado no bioma Cerrado.

An epidemiological survey of all cases of bovine rabies that occurred in Mato Grosso, Brazil, during the years 1996 to 2006 was conducted by way of the analysis of the epidemic curve, by calculating the sum, average, standard deviation, maximum and minimum values of the occurrence of rabies in three ecosystems; statistical analysis between the actual cattle in the three biomes of the state and the number of cases of rabies cattle; and the seasonal distribution and disease situation in the state in relation to the national scene. The data were organized by taking into account the month and year of occurrence of rabies and the geographic region of the origin of the material. The different cities involved were classified according to their location in regions of Cerrado, the Pantanal and the Amazon. It was found that the variation in the number of positive cases of the disease has been increasing in recent years, with the largest number of cases occurring in the Cerrado biome, which showed a trend of annual increase. Beginning in the year 2004 the Cerrado ecosystem passed from the situation of enzootic to epizootic, making it evident that the main bovine rabies problem in the state of Mato Grosso is situated in the Cerrado biome.

Caracterização genética e distribuição geográfica do vírus rábico isolado de bovinos no estado de Mato Grosso, Brasil / Genetic characterization and geographic distribution of rabies virus isolates from cattle in Mato Grosso, Brazil

Foram caracterizados, geneticamente e geograficamente, o sequenciamento parcial da nucleoproteína (gene N) de 53 isolados do vírus da raiva (VR) originários do Estado de Mato Grosso, Brasil. Os isolados de bovinos, que se encontravam no grupo do VR relacionado a morcegos hematófagos, foram posteriormente subdivididos em sete subgrupos genéticos. Estes subgrupos foram distribuídos em regiões de terras planas, com alguns subgrupos separados por formações de pequenas montanhas e hidrografia. Estes resultados indicam que a raiva em bovinos é derivada de diversas variantes regionalmente definidas, o que sugere que sua distribuição geográfica está relacionada as populações de morcegos hematófagos.

A total of 53 rabies virus (RV) isolates originating from cattle in the state of Mato Grosso, Brazil, were genetically characterized. Partial nucleoprotein gene sequences of these isolates were phylogenetically and geographically analyzed. Cattle isolates, which clustered with the vampire bat related RV group, were further subdivided into 7 subgroups. These subgroups were distributed widely in lowland regions, with some subgroups separated from each other by small mountains and hydrographical features. These results indicate that cattle rabies is derived from several regionally-defined variants, which suggests that its geographical distribution is related to that of the vampire bat population.

Exercise may cause myocardial ischemia at the anaerobic threshold in cardiac rehabilitation programs

Myocardial ischemia may occur during an exercise session in cardiac rehabilitation programs. However, it has not been established whether it is elicited when exercise prescription is based on heart rate corresponding to the anaerobic threshold as measured by cardiopulmonary exercise testing. Our objective was to determine the incidence of myocardial ischemia in cardiac rehabilitation programs according to myocardial perfusion SPECT in exercise programs based on the anaerobic threshold. Thirty-nine patients (35 men and 4 women) diagnosed with coronary artery disease by coronary angiography and stress technetium-99m-sestamibi gated SPECT associated with a baseline cardiopulmonary exercise test were assessed. Ages ranged from 45 to 75 years. A second cardiopulmonary exercise test determined training intensity at the anaerobic threshold. Repeat gated-SPECT was obtained after a third cardiopulmonary exercise test at the prescribed workload and heart rate. Myocardial perfusion images were analyzed using a score system of 6.4 at rest, 13.9 at peak stress, and 10.7 during the prescribed exercise (P < 0.05). The presence of myocardial ischemia during exercise was defined as a difference ≥2 between the summed stress score and summed rest score. Accordingly, 25 (64 percent) patients were classified as ischemic and 14 (36 percent) as nonischemic. MIBI-SPECT showed myocardial ischemia during exercise within the anaerobic threshold. The 64 percent prevalence of ischemia observed in the study should not be looked on as representative of the whole population of patients undergoing exercise programs. Changes in patient care and exercise programs were implemented as a result of our finding of ischemia during the prescribed exercise.

Prevalência de rotavírus do grupo A em fezes diarréicas de bezerros de corte em sistema semi-intensivo de produçäo / Prevalence of group A rotavirus in diarrheic feaces of beef calves in semi-intensive production system

Determinou-se a prevalência de rotavírus durante surto de diarréia em bezerros de um rebanho de corte, criado em regime semi-intensivo de produçäo. Analisaram-se, por meio de técnicas de eletroforese em gel de poliacrilamida (EGPA) e ensaio imunoenzimático (kit EIARA - Fiocruz), 69 amostras de fezes de bezerros, entre 30 e 60 dias de idade, colhidas em três estaçöes de pariçäo consecutivas (agosto a novembro/1999, janeiro a abril e agosto a novembro/2000). Pelo EIARA foram detectadas 63,8 por cento (44/69) de amostras positivas. Na primeira estaçäo de pariçäo foi detectado rotavírus em 82,4 por cento (14/17) dos bezerros que apresentaram quadro clínico de diarréia. No ano de 2000 a presença de rotavírus foi detectada em 41,7 por cento (5/12) e 62,5 por cento (25/40) do total de amostras examinadas. A análise do perfil eletroforético do genoma indicou grande diversidade, com quatro eletroferótipos distintos, todos com perfil longo, característico de rotavírus do grupo A.