Antimicrobial, antioxidant activity and phytochemical prospection of Eugenia involucrata DC. leaf extracts / Atividade antimicrobiana, antioxidante e prospecção fitoquímica dos extratos das folhas de Eugenia involucrata DC

Abstract The species Eugenia involucrata DC. is a plant native to Brazil and is traditionally used for intestinal problems, however, little research has documented about its biological potential and phytochemical profile. Thus, the objective of this study was to carry out preliminary phytochemical prospecting, antimicrobial and antioxidant potential of E. involucrata extracts. Using the E. involucrata leaves, aqueous and organic extracts were obtained using the following solvents (ethanol, methanol, hexane, acetone, dichloromethane and ethyl acetate). The phytochemical prospecting detected the presence of saponins, steroids, flavonoids and tannins in the extracts. Ethanolic and methanolic extracts presented antimicrobial activity for most of the bacterial strains tested, as well as for yeast Candida albicans, with concentrations between 3.12 and 50 mg/mL. The ethanolic and metanolic extract presented high free radical sequestration potential (>90%). The methanol extract showed an IC50 value statistically equal to that found for the commercial antioxidant BHT (p <0.05). The crude extracts obtained with ethanol and methanol were the most promising. These results suggest that methanolic, ethanolic and aqueous extracts are a promising source of natural bioactive.

Resumo A espécie Eugenia involucrata DC. é uma planta nativa do Brasil e tradicionalmente utilizada para problemas intestinais, porém, poucas pesquisas documentam sobre seu potencial biológico e perfil fitoquímico. Assim, o objetivo deste trabalho foi realizar uma prospecção fitoquímica preliminar, o potencial antimicrobiano e antioxidante dos extratos de E. involucrata. A partir das folhas de E. involucrata, foram obtidos extratos aquosos e orgânicos com os seguintes solventes (etanol, metanol, hexano, acetona, diclorometano e acetato de etila). A prospecção fitoquímica detectou a presença de saponinas, esteroides, flavonóides e taninos nos extratos. Os extratos etanólico e metanólico apresentaram atividade antimicrobiana para a maioria das cepas bacterianas testadas, assim como para a levedura Candida albicans, com concentrações entre 3,12 e 50 mg/mL. Os extratos etanólico e metanólico apresentaram alto potencial de sequestro de radicais livres (> 90%). O extrato metanólico apresentou IC50 estatisticamente igual ao encontrado para o antioxidante comercial BHT (p <0,05). Os extratos brutos obtidos com etanol e metanol foram os mais promissores. Esses resultados sugerem que os extratos metanólico, etanólico e aquoso são uma fonte promissora de bioativos naturais.

Antimicrobial, antioxidant activity and phytochemical prospection of Eugenia involucrata DC. leaf extracts

Abstract The species Eugenia involucrata DC. is a plant native to Brazil and is traditionally used for intestinal problems, however, little research has documented about its biological potential and phytochemical profile. Thus, the objective of this study was to carry out preliminary phytochemical prospecting, antimicrobial and antioxidant potential of E. involucrata extracts. Using the E. involucrata leaves, aqueous and organic extracts were obtained using the following solvents (ethanol, methanol, hexane, acetone, dichloromethane and ethyl acetate). The phytochemical prospecting detected the presence of saponins, steroids, flavonoids and tannins in the extracts. Ethanolic and methanolic extracts presented antimicrobial activity for most of the bacterial strains tested, as well as for yeast Candida albicans, with concentrations between 3.12 and 50 mg/mL. The ethanolic and metanolic extract presented high free radical sequestration potential (>90%). The methanol extract showed an IC50 value statistically equal to that found for the commercial antioxidant BHT (p 0.05). The crude extracts obtained with ethanol and methanol were the most promising. These results suggest that methanolic, ethanolic and aqueous extracts are a promising source of natural bioactive.

Resumo A espécie Eugenia involucrata DC. é uma planta nativa do Brasil e tradicionalmente utilizada para problemas intestinais, porém, poucas pesquisas documentam sobre seu potencial biológico e perfil fitoquímico. Assim, o objetivo deste trabalho foi realizar uma prospecção fitoquímica preliminar, o potencial antimicrobiano e antioxidante dos extratos de E. involucrata. A partir das folhas de E. involucrata, foram obtidos extratos aquosos e orgânicos com os seguintes solventes (etanol, metanol, hexano, acetona, diclorometano e acetato de etila). A prospecção fitoquímica detectou a presença de saponinas, esteroides, flavonóides e taninos nos extratos. Os extratos etanólico e metanólico apresentaram atividade antimicrobiana para a maioria das cepas bacterianas testadas, assim como para a levedura Candida albicans, com concentrações entre 3,12 e 50 mg/mL. Os extratos etanólico e metanólico apresentaram alto potencial de sequestro de radicais livres (> 90%). O extrato metanólico apresentou IC50 estatisticamente igual ao encontrado para o antioxidante comercial BHT (p 0,05). Os extratos brutos obtidos com etanol e metanol foram os mais promissores. Esses resultados sugerem que os extratos metanólico, etanólico e aquoso são uma fonte promissora de bioativos naturais.

A method to assess heart rate variability in neonate rats: validation in normotensive and hypertensive animals

Several studies have focused on the heart rate variability (HRV) of murine species, while studies discussing HRV in murine neonates and infants remain scarce, since recording hemodynamic signals through invasive methods in small animals has been found to be quite challenging. Thus, this study aimed at describing and validating a novel method to assess HRV in newborn rats. An electrocardiogram (ECG) system was used to determine RR intervals in awake newborns and evaluate HRV in normotensive (Wistar) and hypertensive (SHR) neonate rats. After birth, ECG was recorded in the awake newborns, and they were allowed to rest on a heated surface, restricted only by the weight of the adhesive ECG electrodes. The electrodes were cut and adapted to provide more comfort to the animal, and gently placed on the newborn's skin. RR intervals were recorded over a 30-min period using an ECG system together with LabChart software (4 KHz). Three sequences of 5 min each from the ECG recording period were analyzed in time and frequency domains, using CardioSeries software. ECG data resulted in a clearly interpretable signal that was used to generate an RR interval sequence through time for the analysis of HRV. SHR neonates presented increased cardiac sympathovagal balance compared to Wistar neonates (low frequency/high frequency: 3.85±0.71 vs 0.90±0.09). In conclusion, the ECG setup here described may be used to record RR intervals to assess HRV in neonate rats, thus detecting early impairment of HRV in hypertensive newborns.

Low level laser therapy for reducing pain in rheumatoid arthritis and osteoarthritis: a systematic review / Terapia Laser de Baixa Intensidade para a redução da dor em artrite reumatoide e osteoatrite: uma revisão sistemática / Terapia con Láser de baja intensidad para reducir el dolor en la artritis reumatoide y la osteoartritis: una revisión sistemática

Abstract Introduction: Treatments for rheumatoid arthritis (RA) and osteoarthritis (OA) can reduce, modulate inflammation, and reduce deformities. Low-Level Laser Therapy is a biomodulator and may aid in the clinical picture of these conditions. Objective: To analyze the parameters most frequently used to determine the responses of patients with RA and OA in controlled and uncontrolled clinical trials. Method: This is a systematic review with search of articles in English, Portuguese and Spanish in PUBMED, SCOPUS, LILACS and Web of SCIENCE, of articles published between 2006 and 2018. MeSH terms were used. Inclusion criteria: evaluation of LLLT in the evaluations, evaluation and evaluation of the period, controlled and uncontrolled clinical trials, full publications. The base date of the energy dosimetry and the analysis of mean, median and mode of energy per point and energy per treatment. Results: Three articles on RA and 16 on OA were included in this study. Regarding dosimetry, it was one of the most recent of the pain, being this one with a greater energy dose. In OA, most of the articles presented are of importance, with variability in the dosage applied. Conclusion: There are several reports for patient studies purposes, mainly with doses of 6 J per point and 48 J. In the joints affected with OA and AR, it would be important to publish more scientific articles with better methodological quality and description of dosimetry.

Resumo Introdução: Os tratamentos para artrite reumatoide (AR) e osteoartrite (OA) devem diminuir dor, modular inflamação e diminuir deformidades. O LLLT é um biomodulador e pode auxiliar no quadro clínico destas. Objetivo: Analisar os parâmetros mais comumente utilizados para a diminuição das respostas de dor em pacientes com AR e OA em ensaios clínicos controlados e não controlados. Método: Trata-se de uma revisão sistemática com busca de artigos em inglês, português e espanhol na: PUBMED, SCOPUS, LILACS e Web of SCIENCE, entre 2006 a 2018. Foram utilizados os descritores MESH. Critérios de inclusão: avaliação da eficácia do LLLT nestas populações, verificação da dor antes e depois do tratamento, ensaios clínicos controlados e não controlados, publicados integralmente. Foi realizado o cálculo da dosimetria do laser e análise de média, mediana e moda da energia por ponto e energia por tratamento. Resultados: Incluídos neste estudo 3 artigos de AR e 16 de OA. Em relação à dosimetria, apenas um AR apresentou redução da dor, sendo este com a maior dose de energia. Já na OA, a maioria dos artigos apresentou diminuição da dor, e com variabilidade na dosagem aplicada. Alguns tiveram poucos parâmetros apresentados, tornando impossível calcular a dosimetria. Conclusão: há evidências científicas para reduzir a dor relatada em pacientes com OA de joelho, principalmente com doses de 6 J por ponto e 48 J aplicadas no tratamento diário. Para as outras articulações afetadas com OA e AR, seria importante a publicação de mais artigos científicos com melhor qualidade metodológica e descrição da dosimetria.

Resumen Introducción: Los tratamientos para la artritis reumatoide (AR) y la osteoartritis (OA) pueden reducirse, modular, encender y disminuir las deformidades. El LLLT es un biomodulador y puede auxiliar en el cuadro clínico de éstas. Objetivo: Analizar los parámetros más frecuentemente utilizados para determinar las respuestas de pacientes con AR y OA en ensayos clínicos controlados y no controlados. Método: Se trata de una revisión sistemática y la búsqueda de artículos en Inglés, portugués y español en: PubMed, SCOPUS, se utilizaron LILACS y Web of Science, entre 2006 y 2018. Los descriptores de malla. Criterios de inclusión: evaluación de la LLLT en las evaluaciones, evaluación y evaluación del período, ensayos clínicos controlados y no controlados, íntegramente. La fecha base de la dosimetría de energía y el análisis de media, media y moda de la energía por punto y energía por tratamiento. Resultados: Incluido en este estudio 3 artículos de AR y 16 de OA. En cuanto a la dosimetría, fue uno de los más recientes del dolor, siendo éste con una dosis mayor de energía. En la OA, la mayoría de los artículos presentados son de importancia, con la variabilidad en la dosificación aplicada. Se han tenido pocos pocos parámetros parámetros parámetros presentados presentados. Conclusión: hay una serie de informes para fines de estudios con pacientes, principalmente con dosis de 6 J por punto y 48 J. Las articulaciones afectadas con OA y AR, sería importante la publicación de más artículos científicos con mejor calidad metodológica y descripción de dosimetría.

Essential oil of Pterodon polygalaeflorus Benth attenuates nociception in mice

Essential oils (EO) are volatile liquids responsible for the aroma of plants. Pterodon polygalaeflorus seeds have received widespread use in folk medicine for the treatment of inflammatory diseases. For this reason and because Pterodon polygalaeflorus seeds have great EO content, which is frequently pharmacologically active, the present study aimed to evaluate the antinociceptive effect of EO from Pterodon polygalaeflorus (EOPPgfl) and its acute toxic effects. The EEOPPgfl sample, which was extracted by steam distillation of the seeds, had a yield of 2.4% of the seeds weight and had, as major constituents, beta-elemene (48.19%), trans-caryophyllene (19.51%), and epi-bicyclosesquiphellandrene (12.24%). The EOPPgfl sample showed mild acute toxicity and its calculated median lethal dose (LD50) was 3.38 g/kg. EOPPgfl (20-60 mg/kg) showed antinociceptive activity as evidenced by several tests and inhibited writhing induced by acetic acid. The maximum effect was obtained with the 30 mg/kg dose and at 60 min after its administration. EOPPgfl also decreased formalin-induced nociception, as verified by the inhibition of the first and second phase of the formalin test. At 30 mg/kg, EOPPgfl also decreased thermally stimulated nociception. Nociception may be related to inflammatory and antiedematogenic activity and at doses ranging 10-100 mg/kg, EOPPgfl blocked dextran- and carrageenan-induced edema. The results demonstrated that EOPPgfl presented, at doses approximately 100 times smaller than LD50, an antinociceptive effect that probably was due to anti-inflammatory activities.

Substâncias fenólicas, flavonoides e capacidade antioxidante em erveiras sob diferentes coberturas do solo e sombreamentos / Phenolic Substances, Flavonoids, and Antioxidant Capacity in Herbs under Different Soil Covers and Shadings

RESUMO Objetivou neste trabalho analisar a concentração de polifenóis totais, flavonoides e capacidade antioxidantes por meio dos métodos ABTS e FRAP, em erveiras jovens cultivadas em solo coberto e com sombreamento. Mudas de erva-mate foram submetidas aos tratamentos com solos cobertos e desnudos em sombreamentos de 0, 18, 35 e 50%, após sete e onze meses de cultivo, verão e outono, respectivamente. Foram coletadas folhas maduras para realização dos extratos utilizados para as análises de compostos fenólicos por meio da reação de oxirredução com reagente de Folin-Ciocalteu, flavonoides por método colorimétrico e capacidade antioxidante por frente ao radical ABTS e poder de redução do ferro (FRAP). O teor de compostos fenólicos foi maior nos tratamentos com 35% de sombreamento em comparação ao grupo dos flavonoides e capacidade antioxidante, o que foi demonstrado pela correlação do sombreamento com estes parametros. No método FRAP a maior correlação demonstra que o outono é a melhor época de colheita por apresentar maior concentração de compostos fitoquímicos. Ficou evidente a influência positiva dos sombreamentos (35 e 50%) no teor de compostos fenólicos e flavonoides relacionados à capacidade antioxidante e à qualidade da erva-mate para atender ao mercado consumidor.

ABSTRACT This study aimed to analyze the concentration of total polyphenols, flavonoids, and antioxidant capacity, by the methods ABTS and FRAP, in young herbs grown in covered and shaded soil. Yerba mate seedlings were subjected to treatment with bare and covered soils in shadings of 0, 18, 35, and 50%, after seven and eleven months cultivation, summer and fall, respectively. Mature leaves were collected to perform the analyses of phenolic compounds by redox reaction with Folin-Ciocalteu reagent, of flavonoids by colorimetric method, and of antioxidant capacity by ABTS radical and FRAP (iron reduction method). The content of phenolic compounds was higher in the treatments with 35% shading in correlation with the group of flavonoids and antioxidant capacity. In the FRAP method, the higher correlation shows that autumn is the best time to harvest because of the higher concentration of phytochemical compounds. The positive influence of shading (35 and 50%) was evident in the content of phenolic compounds and flavonoids related to antioxidant capacity and better quality of yerba mate to meet the consumer market.

Relação entre obesidade, insulina plasmática e posicionamento da falange distal em equinos da raça Crioula / Relationship between obesity, insulin and placement of plasma in equine distal phalanx of breed creole

Verificaram-se características radiográficas do posicionamento da falange distal em relação ao estojo córneo em equinos da raça Crioula e correlacionaram-se tais achados com sinais de obesidade e concentrações plasmáticas de insulina. Consideraram-se dois grupos com base no escore da condição corporal (ECC). A média de idade entre os grupos foi de 9,2±5,8 anos em indivíduos normais (ECC 5-7) e de 10,3±3,7 anos em obesos (ECC 8-9). Diferenças estatísticas entre grupos ocorreram para valores de peso, perímetro torácico, escore da condição corporal, escore da crista do pescoço, comprimento do pescoço, circunferência do pescoço em três regiões referentes a 25%, 50% e 75% do comprimento do pescoço e entre o posicionamento da falange distal em relação ao estojo córneo. O ângulo formado entre as superfícies dorsais de falange distal e casco (ângulo de rotação) correlacionou-se estatisticamente com ECC (r = 0,30; P=0,02) e com peso e perímetro torácico (r = 0,50; P<0,01). Quanto à concentração de insulina plasmática, foram encontradas correlações positivas com ECP (r = 0,40; P<0,01) e com ângulo de rotação (r = 0,23; P = 0,08), e correlação negativa com idade (r = -0,42; P<0,01). Em oito animais (27%) o ângulo de rotação foi maior que 2 graus; destes, 25% eram normais e 75%, obesos (P<0,05). Aparentemente, em equinos da raça Crioula, características morfométricas no casco diferiram dos padrões internacionais obtidos de outras raças. A obesidade interferiu na relação espacial da falange distal com o estojo córneo, indicando que os animais obesos dessa raça são mais propensos a desenvolver laminite.

The radiographic relationship of the distal phalanx with the hoof capsule was verified in Creole breed horses and these findings correlated with signs of obesity and insulin blood levels. Horses were divided in two groups based on their body score condition (ECC). The average age was 9.2±5.8 years in normal (ECC 5-7) and 10.3±3.7 years in the obese group (ECC 8-9). Statistical differences between groups (P<0.05) were detected for values of weight, thoracic girth, body score condition, neck score (ECP), neck length, neck circumference into three regions based on 25% (P25), 50% (P50) and 75% (P75) of the length of the neck and phalanx and hoof capsule relationship. The angle between the dorsal aspects of distal phalanx and hoof wall (rotation angle) statistically correlated with ECC (r = 0.30; P=0.02) and with weight and thoracic girth (r = 0.50; P<0.01). Plasma insulin concentrations positively correlated with ECP (r = 0.40, P <0.01) and rotation angle (r = 0.23, P = 0.08) and negatively with age (r = -0.42, P <.01). In eight animals (27%) rotation angle was greater than 2 degrees, of which 25% were normal and obese 75% (P <0.05). Apparently in Creole horses, morphometric characteristics of the hoof differ from international standards obtained from other breeds. Obesity interfered with the spatial relationship of the distal phalanx with the hoof capsule, indicating that obese animals of this breed are more likely to develop laminitis.

Mental rotation of anthropoid hands: a chronometric study

It has been shown that mental rotation of objects and human body parts is processed differently in the human brain. But what about body parts belonging to other primates? Does our brain process this information like any other object or does it instead maximize the structural similarities with our homologous body parts? We tried to answer this question by measuring the manual reaction time (MRT) of human participants discriminating the handedness of drawings representing the hands of four anthropoid primates (orangutan, chimpanzee, gorilla, and human). Twenty-four right-handed volunteers (13 males and 11 females) were instructed to judge the handedness of a hand drawing in palm view by pressing a left/right key. The orientation of hand drawings varied from 0° (fingers upwards) to 90° lateral (fingers pointing away from the midline), 180° (fingers downwards) and 90° medial (finger towards the midline). The results showed an effect of rotation angle (F(3, 69) = 19.57, P < 0.001), but not of hand identity, on MRTs. Moreover, for all hand drawings, a medial rotation elicited shorter MRTs than a lateral rotation (960 and 1169 ms, respectively, P < 0.05). This result has been previously observed for drawings of the human hand and related to biomechanical constraints of movement performance. Our findings indicate that anthropoid hands are essentially equivalent stimuli for handedness recognition. Since the task involves mentally simulating the posture and rotation of the hands, we wondered if "mirror neurons" could be involved in establishing the motor equivalence between the stimuli and the participants' own hands.

Otimização do processo de extração de própolis através da verificação da atividade antimicrobiana

A própolis, também conhecida como "cola de abelhas" possui atividade antimicrobiana, antiinflamatória, anestésica e imunoestimulante. Amostras de diferentes origens geográficas podem apresentar composição química bem diferente, porém continuam sendo responsáveis por atividade antibacteriana, inclusive contra cepas multirresistentes de Staphylococcus aureus. Este trabalho teve por objetivo otimizar o processo de extração visando determinar a melhor alcoolatura para a obtenção de tinturas de própolis com maior atividade antimicrobiana. Amostras de própolis foram extraídas com diferentes misturas hidroalcoólicas, variando o teor de etanol entre 90 por cento a água pura, obtendo assim 11 tinturas diferentes. Todas as tinturas foram aplicadas em discos de papel, secos e colocados sobre a superfície de placas contendo ágar inoculado com cepa multirresistente de S. aureus. As placas foram incubadas e avaliadas no dia seguinte quanto à presença de halos de inibição. Os melhores resultados foram obtidos com extratos contendo 50 a 90 por cento de etanol. O trabalho sugere a utilização da alcoolatura de 70 por cento como a ideal para a obtenção de tinturas de própolis.

Bee glue (propolis) possesses antimicrobial, antiinflammatory, anaesthetic and immunostimulating activities. Propolis composition is extraordinary variable; samples from different geographic origin may posse's entirely different chemical composition. In different propolis samples, various substance combinations are responsible for the antibacterial activity of the bee glue. The aim of the study was to evaluate the best ethanolic extract concentration using Staphylococcus aureus multiresistant. Ethanol extracts from different concentration (10 to 90 percent) were applied on paper disks, dried, and put on the surface of nutrient agar plates with the overlay containing S. aureus. The plates were then incubated and evaluated the next day for the presence of inhibition zones. The best result was verified among 60 to 90 percent of ethanolic extract of propolis. This paper suggests the use of the alcool 70 percent as the ideal for the extration of propolis.

Application of hazard analysis critical control point (HACCP) as a possible control measure for Opisthorchis viverrini infection in cultured carp (Puntius gonionotus).

Opisthorchiasis due to Opisthorchis viverrini and transmitted through infected freshwater cyprinoid fish (carps) affects more than 8 million people in Thailand, People's Democratic Republic of Lao, and Vietnam. The Hazard Analysis Critical Control Point (HACCP)-concept has been recommended by FAO and WHO to be included in programs to control foodborne trematode infections (FBT). HACCP is a multifactorial approach to control food hazards through surveillance of diseases, foods, and operations and education. This study describes the first attempt to apply HACCP to the prevention and control of Opisthorchis viverrini in pond culture carp (Puntius gonionotus). The experiment was designed and carried out by a multidisciplinary "HACCP team" including experts in the field of public health, parasitology, epidemiology, aquaculture, fisheries extension and fish inspection. The investigation was performed in two fish ponds in the District of Sun Pa Tong, Chiang Mai, Thailand. In the experimental pond, fish was cultured according to HACCP principles and compared with the control pond, which followed conventional aquaculture practices. Water supply to the pond, fish fry, fish feed and pond conditions during the growing period were identified as critical control points (CCPs). Hazards were identified and analyzed, as well as control measures, critical limits, monitoring procedures, corrective actions, and record keeping developed for each one of the above CCPs. Complete pond preparation, particularly aiming to eliminate contamination of pond water with O. viverrini eggs, fish infected with parasite meacercariae and the first intermediate host (Bithynia spp), was conducted. After the pond was filled with water, O. viverrini metacercaria-free fry were released into the pond. The preliminary results obtained indicate that HACCP-based principles applied to carp pond culture could be used as a strategy to prevent and control O. viverrini. Further studies should be undertaken aiming to confirm these preliminary results.

Mutantes con alta producción de butanol de una cepa de Clostridium solventogénica aislada de alpechín / [Mutants with high butanol production from a strain of solventogenic Clostridium isolated from olive black-water]

Butanol high producing mutants of a solventogenic Clostridium sp. capable of degrading olive black water, were selected according to ethanol or butanol resistance after treatment with N-methyl-N’-nitro-N-nitrosoguanidine. Mutants were quickly screened from isolated colonies and then characterized in standard culture conditions.