Ferulic acid as major antioxidant phenolic compound of the Tetragonisca angustula honey collected in Vera Cruz - Itaparica Island, Bahia, Brazil

Abstract The antioxidant activity of Tetragonisca angustula honey (TAH) and its ethanolic extract (TAEE) were investigated. The total levels of phenolic (TPC) and flavonoids (TFC) were also evaluated. The results for TPC were 19.91 ± 0.38 and 29.37 ± 1.82 mg GAE g-1 and for TFC 0.20 ± 0.02 and 0.14 ± 0.01 mg QE g-1 of TAH and TAEE, respectively. Antioxidant activities were 73.29 ± 0.49% and 93.36 ± 0.27% in the DPPH assay and 71.73 ± 4.07% and 97.86 ± 0.35% in ABTS+ for TAH and TAEE, respectively. The total reducing activity was determined by the method of reducing power (PR) and iron ion (Fe III) and the results varied in PR from 151.7 ± 25.7 and 230.7 ± 25.2 mg GAE L-1, for TAH and TAEE respectively and for (Fe III) in EC50 0.284 in TAEE and 0.687 in TAH. Chemical analysis by HPLC-DAD of the ethanolic extract (TAEE) revealed the presence of ferulic acid as majority phenolic component in the extract. The 1H NMR analysis confirmed this structure and showed the also presence of glucose, citric acid, succinic acid, proline and hydrocarbon derivatives. In addition, the botanical origin was also investigated and showed a multifloral characteristic, having found 19 pollen types with a botanical predominance of the Anacardiaceae family, with Tapirira pollen occurring as predominant (42.6%) and Schinus as secondary (25.7%). The results showed that T. angustula honey is an interesting source of antioxidant phenolic compounds due to its floral origin and can act as a protector of human health when consumed.

Resumo A atividade antioxidante do mel de Tetragonisca angustula (TAH) e seu extrato etanólico (TAEE) foram investigados. Os níveis totais de fenólicos (TPC) e flavonóides (TFC) também foram avaliados. Os resultados para TPC foram 19,91 ± 0,38 e 29,37 ± 1,82 mg GAE g-1 e para TFC 0,20 ± 0,02 e 0,14 ± 0,01 mg QE g-1 de TAH e TAEE, respectivamente. As atividades antioxidantes foram 73,29 ± 0,49% e 93,36 ± 0,27% no ensaio DPPH e 71,73 ± 4,07% e 97,86 ± 0,35% no ABTS+ para TAH e TAEE, respectivamente. A atividade redutora total foi determinada pelo método de poder redutor (PR) e íon ferrico (Fe III) e os resultados variaram em PR de 151,7 ± 25,7 e 230,7 ± 25,2 mg GAE L-1, para TAH e TAEE respectivamente e para (Fe III) em EC50 0,284 em TAEE e 0,687 em TAH. A análise química por HPLC-DAD do extrato etanólico (TAEE) revelou a presença de ácido ferúlico como componente majoritário no extrato. A análise de RMN 1H confirmou esta estrutura e mostrou a presença de glicose, ácido cítrico, ácido succínico, prolina e derivados de hidrocarbonetos no TAEE. Além disso, a origem botânica também foi investigada e apresentou característica multifloral, tendo encontrado 19 tipos polínicos com predomínio botânico da família Anacardiaceae, sendo o pólen Tapirira predominante (42,6%) e o Schinus secundário (25,7%). Os resultados mostraram que o mel de T. angustula é uma interessante fonte de compostos fenólicos antioxidantes devido à sua origem floral e pode atuar como protetor da saúde humana quando consumido.

Ferulic acid as major antioxidant phenolic compound of the Tetragonisca angustula honey collected in Vera Cruz - Itaparica Island, Bahia, Brazil / Ácido ferúlico como principal composto fenólico antioxidante do mel de Tetragonisca angustula coletado em Vera Cruz - Ilha de Itaparica, Bahia, Brasil

Abstract The antioxidant activity of Tetragonisca angustula honey (TAH) and its ethanolic extract (TAEE) were investigated. The total levels of phenolic (TPC) and flavonoids (TFC) were also evaluated. The results for TPC were 19.91 ± 0.38 and 29.37 ± 1.82 mg GAE g-1 and for TFC 0.20 ± 0.02 and 0.14 ± 0.01 mg QE g-1 of TAH and TAEE, respectively. Antioxidant activities were 73.29 ± 0.49% and 93.36 ± 0.27% in the DPPH● assay and 71.73 ± 4.07% and 97.86 ± 0.35% in ABTS●+ for TAH and TAEE, respectively. The total reducing activity was determined by the method of reducing power (PR) and iron ion (Fe III) and the results varied in PR from 151.7 ± 25.7 and 230.7 ± 25.2 mg GAE L-1, for TAH and TAEE respectively and for (Fe III) in EC50 0.284 in TAEE and 0.687 in TAH. Chemical analysis by HPLC-DAD of the ethanolic extract (TAEE) revealed the presence of ferulic acid as majority phenolic component in the extract. The 1H NMR analysis confirmed this structure and showed the also presence of glucose, citric acid, succinic acid, proline and hydrocarbon derivatives. In addition, the botanical origin was also investigated and showed a multifloral characteristic, having found 19 pollen types with a botanical predominance of the Anacardiaceae family, with Tapirira pollen occurring as predominant (42.6%) and Schinus as secondary (25.7%). The results showed that T. angustula honey is an interesting source of antioxidant phenolic compounds due to its floral origin and can act as a protector of human health when consumed.

Resumo A atividade antioxidante do mel de Tetragonisca angustula (TAH) e seu extrato etanólico (TAEE) foram investigados. Os níveis totais de fenólicos (TPC) e flavonóides (TFC) também foram avaliados. Os resultados para TPC foram 19,91 ± 0,38 e 29,37 ± 1,82 mg GAE g-1 e para TFC 0,20 ± 0,02 e 0,14 ± 0,01 mg QE g-1 de TAH e TAEE, respectivamente. As atividades antioxidantes foram 73,29 ± 0,49% e 93,36 ± 0,27% no ensaio DPPH● e 71,73 ± 4,07% e 97,86 ± 0,35% no ABTS●+ para TAH e TAEE, respectivamente. A atividade redutora total foi determinada pelo método de poder redutor (PR) e íon ferrico (Fe III) e os resultados variaram em PR de 151,7 ± 25,7 e 230,7 ± 25,2 mg GAE L-1, para TAH e TAEE respectivamente e para (Fe III) em EC50 0,284 em TAEE e 0,687 em TAH. A análise química por HPLC-DAD do extrato etanólico (TAEE) revelou a presença de ácido ferúlico como componente majoritário no extrato. A análise de RMN 1H confirmou esta estrutura e mostrou a presença de glicose, ácido cítrico, ácido succínico, prolina e derivados de hidrocarbonetos no TAEE. Além disso, a origem botânica também foi investigada e apresentou característica multifloral, tendo encontrado 19 tipos polínicos com predomínio botânico da família Anacardiaceae, sendo o pólen Tapirira predominante (42,6%) e o Schinus secundário (25,7%). Os resultados mostraram que o mel de T. angustula é uma interessante fonte de compostos fenólicos antioxidantes devido à sua origem floral e pode atuar como protetor da saúde humana quando consumido.

Physical effectiveness of corn silage fractions stratified with the Penn State Particle Separator for lactating dairy cows.

This study evaluated the physical effectiveness of whole-plant corn silage (CS) particles stratified with the Penn State Particle Separator, composed of 19- and 8-mm-diameter sieves and a pan, for lactating dairy cows. Eight Holstein cows (27.6 ± 2.8 kg/d of milk, 611 ± 74 kg body weight; 152 ± 83 d in milk) were assigned to two 4 × 4 Latin squares (22-d periods, 16-d adaptation), where one square was formed with rumen-cannulated cows. Three CS particle fractions were manually isolated using the 8- and 19-mm diameter sieves and re-ensiled in 200-L drums. The 4 experimental diets were (% dry matter): (1) CON (control): 17% forage neutral detergent fiber (NDF) from CS (basal roughage), 31.5% starch, and 31.9% NDF; (2) PSPan: 17% forage NDF from CS + 9% NDF from CS particles <8 mm, 25.9% starch, and 37.9% NDF; (3) PS8: 17% forage NDF from CS + 9% NDF from CS particles 8 to 19 mm, 25.5% starch, and 38.3% NDF; and (4) PS19: 17% forage NDF from CS + 9% NDF from CS particles >19 mm, 24.9% starch, and 38.8% NDF. Cows fed PS8 had greater dry matter intake and energy-corrected milk yield (22.4 and 26.9 kg/d, respectively) than cows fed CON (20.8 and 24.7 kg/d) and PS19 (21.2 and 24.8 kg/d), but no difference was detected between PSPan (21.6 and 25.8 kg/d) and other treatments. Milk fat concentration was greater for PS8 than CON, with intermediate values for PSPan and PS19. Milk fat yield was greater for cows fed PS8 than CON and PS19, and cows fed PSPan secreted more fat than CON cows but were not different from cows fed the other 2 diets. Cows fed CON had a lower meal frequency than cows fed PSPan, shorter meal and rumination times than PS8, and greater meal size and lower rates of rumination and chewing than the other 3 diets. Total chewing per unit of NDF was higher for PS8 than PSPan, although neither treatment differed from CON or PS19. Cows fed PS19 had higher refusal of feed particles >19 mm than cows fed CON and PSPan. The refusal of dietary NDF and undigested NDF in favor of starch were all greater for PS19 than on the other treatments. Cows fed PS19 had a greater proportion of the swallowed bolus and rumen digesta with particles >19 mm than the other 3 diets. Cows fed CON had the lowest ruminal pH and greatest lactate concentration relative to the other 3 diets. Plasma lipopolysaccharide was higher for cows fed CON and PSPan than for cows fed PS8 and PS19, and serum d-lactate tended to be lower on PSPan than for CON and PS8. In summary, the inclusion of CS fractions in a low-forage fiber diet (CON) reduced signs of ruminal acidosis. Compared with CS NDF <8 and >19 mm, CS NDF with 8- to 19-mm length promoted better rumen health and performance of dairy cows. These results highlight the importance of adjusting CS harvest and formulating dairy diets based on the proportion of particles retained between the 8- and 19-mm sieves.

Ferulic acid as major antioxidant phenolic compound of the Tetragonisca angustula honey collected in Vera Cruz - Itaparica Island, Bahia, Brazil.

The antioxidant activity of Tetragonisca angustula honey (TAH) and its ethanolic extract (TAEE) were investigated. The total levels of phenolic (TPC) and flavonoids (TFC) were also evaluated. The results for TPC were 19.91 ± 0.38 and 29.37 ± 1.82 mg GAE g-1 and for TFC 0.20 ± 0.02 and 0.14 ± 0.01 mg QE g-1 of TAH and TAEE, respectively. Antioxidant activities were 73.29 ± 0.49% and 93.36 ± 0.27% in the DPPH● assay and 71.73 ± 4.07% and 97.86 ± 0.35% in ABTS●+ for TAH and TAEE, respectively. The total reducing activity was determined by the method of reducing power (PR) and iron ion (Fe III) and the results varied in PR from 151.7 ± 25.7 and 230.7 ± 25.2 mg GAE L-1, for TAH and TAEE respectively and for (Fe III) in EC50 0.284 in TAEE and 0.687 in TAH. Chemical analysis by HPLC-DAD of the ethanolic extract (TAEE) revealed the presence of ferulic acid as majority phenolic component in the extract. The 1H NMR analysis confirmed this structure and showed the also presence of glucose, citric acid, succinic acid, proline and hydrocarbon derivatives. In addition, the botanical origin was also investigated and showed a multifloral characteristic, having found 19 pollen types with a botanical predominance of the Anacardiaceae family, with Tapirira pollen occurring as predominant (42.6%) and Schinus as secondary (25.7%). The results showed that T. angustula honey is an interesting source of antioxidant phenolic compounds due to its floral origin and can act as a protector of human health when consumed.

Zap1 is required for Candida glabrata response to fluconazole.

The increasing prevalence of fluconazole-resistant clinical isolates of Candida spp. strongly hinders the widespread use of the drug. To tackle this problem, great efforts have been made to fully understand the fungal response to fluconazole. In this work, we show that the role of Zap1 in Candida glabrata goes beyond regulating yeast adaptation to zinc deficiency. In line with our previous observation that deletion of ZAP1 makes yeast cells more sensitive to fluconazole, we found that the mutant CgΔzap1 accumulates higher levels of the drug, which correlates well with its lower levels of ergosterol. Surprisingly, Zap1 is a negative regulator of the drug efflux transporter gene CDR1 and of its regulator, PDR1. The apparent paradox of drug accumulation in cells where genes encoding transporters relevant for drug extrusion are being overexpressed led us to postulate that their activity could be impaired. In agreement, Zap1-depleted cells present, in addition to decreased ergosterol levels, an altered composition of membrane phospholipids, which together should impact membrane function and impair the detoxification of fluconazole. Overall, our study brings to light Zap1 as an important hub in Candida glabrata response to fluconazole.

Effect of sealing strategy on the feeding value of corn silage for growing dairy heifers.

Our objective was to compare the performance of dairy heifers fed diets based on whole-plant corn silage stored in bunker silos sealed with either standard polyethylene film (white-on-black, actual thickness 121 ± 3.1 µm) covering the top surface, held in place with rows of tires every 3 m (PE) or an oxygen barrier system comprised of an ethylene-vinyl alcohol film (actual thickness 46.7 ± 2.5 µm) lining side walls and covering the silage, protected with a woven anti-UV cover and gravel bags placed around the edges and every 3 m across the silo (OB). Whole-plant corn was mechanically harvested at 39% dry matter (DM), packed in bunker silos, and sealed with PE or OB covering methods. After 6 mo of storage, silos were opened and fed to 26 Holstein heifers (260 ± 89.1 kg of shrunk body weight) for 60 d. Heifers were blocked by initial weight (13 blocks with 2 heifers each block) and housed in individual pens. Diets contained (on a DM basis) 80% corn silage (PE or OB), 17.5% soybean meal, and 2.5% mineral mix. Dry matter intake was measured daily, whereas shrunk body weight, hip height, heart girth, and body condition score were measured at the beginning and end of the experiment. Feeding behavior was recorded on d 24 and 46, and total-tract digestibility was measured from d 26 to 30 and 48 to 52. Data of intake, feeding behavior, and digestibility were averaged by animal for the whole feeding period before the statistical analysis. Data of animal performance were analyzed as a randomized complete block design. Initial shrunk body weight was used as a covariate for analyses of intake and body measures. During feed-out, silage quality was also assessed at the top (15 cm depth from upper surface) and bottom layer (135 cm depth from upper surface) and analyzed as a split-plot design. Silage stored under the OB sealing system had less yeast, mold, and NH3-N, and more lactic acid and ethanol-soluble carbohydrates. An interaction between sealing strategy and silo layer showed that OB silage had lower values of temperature, pH, anaerobic spores, acetic acid, and DM loss, and greater in vitro DM digestibility and aerobic stability, especially in the top layer. The proportion of inedible silage was lower in OB than in PE treatment (0.82 vs. 4.00% DM). Total-tract digestibility was similar between treatments, but animals that received the OB diet had higher DM intake by approximately 9% (9.39 vs. 10.20 kg/d) due to a faster eating rate and a greater number of meals per day. Therefore, OB treatment increased the digestible energy intake by 8% (26.3 vs. 28.3 Mcal/d) and average daily gain by 12% (1.08 vs. 1.21 kg/d). Body condition score change was similar between treatments, but heifers fed OB had greater heart girth and tended to have higher hip height. In conclusion, replacing a standard PE film with an OB sealing system improved silage conservation and performance of growing dairy heifers.

Seven years of evaluation of ectoparasites and vector-borne pathogens among ring-tailed coatis in an urban park in southeastern Brazil.

Wild animals have been recognized as potential reservoirs of vector-borne pathogens. Proximity between these animals and urban areas increases the need to know which pathogens these are and whether they can infect domestic animals and humans. In Mangabeiras Municipal Park in Belo Horizonte, Brazil, coatis live near the urban area, which is mainly occupied by human residents and their domestic animals. Therefore, the objective of this study was to detect, through molecular and direct methods, the presence of ectoparasites and hemoparasites in coatis. A total of 216 samples were collected, of which 209 samples were from first-captures and seven were from recaptures. The following parasites were found: ticks of the genus Amblyomma, lice of the species Neotrichodectes pallidus and fleas of the species Rhopalopsyllus lutzi lutzi and Ctenocephalides felis felis. All the samples were negative for the family Anaplasmataceae and the species Leishmania sp. and Trypanosoma cruzi. The hemoparasites Trypanosoma evansi, Hepatozoon procyonis, Babesia sp. and Sarcocystis neurona were found. The area of the present study is not endemic for T. evansi, which therefore suggests that these coatis may be acting as reservoirs or sentinels of this parasite. This finding is of great epidemiological importance and should be investigated more closely. Thus, this study showed that there is a great variety of pathogens in the park that transit among coatis and, probably, among other animals that inhabit or live close to the park.

Translation into Brazilian Portuguese, cross-cultural adaptation, reliability and validation of the Functional Ambulation Classification for the categorization of ambulation following a stroke in a clinical setting. / Traducción al portugués brasileño, adaptación transcultural, fiabilidad y validación de la Functional Ambulation Classification para la categorización de deambulación postictus en un entorno clínico.

INTRODUCTION: The Functional Ambulation Classification (FAC) is the only scale that classifies the gait of individuals with multiple sclerosis and stroke victims into ambulation categories. However, the FAC is only available in English and studies conducted in Brazil have used the FAC without an adequate translation and analysis of its measurement properties. AIM: To translate, cross-culturally adapt the FAC to Brazilian Portuguese, test its reliability and concurrent validity on stroke survivors. PATIENTS AND METHODS: The translation and cross-cultural adaptation involved six steps. Inter-observer reliability was tested with five physiotherapists who watched videos of the gait of the stroke survivors and watched a second time after at least one week for the determination of intra-observer reliability. Concurrent validity was determined by correlating the FAC with the result of the 10-meter Walk Test (10mWT). RESULTS: Intra-observer reliability ranged from kappa 0.680 to 0.952 (p = 0.001) and inter-observer reliability ranged from kappa 0.517 to 0.794 (p = 0.001). The correlation between the FAC and 10mWT was rs = 0.771 (p = 0.001). CONCLUSION: The translation, cross-cultural adaptation and measurement properties demonstrated that the FAC is a valid, reliable clinical measure for the categorization of ambulation in the Brazilian population of stroke survivors in a clinical setting.

TITLE: Traducción al portugués brasileño, adaptación transcultural, fiabilidad y validación de la Functional Ambulation Classification para la categorización de deambulación postictus en un entorno clínico.Introducción. La Functional Ambulation Classification (FAC) es la única escala que clasifica la marcha de individuos con esclerosis múltiple e ictus en categorías de deambulación. Sin embargo, la FAC sólo está disponible en la versión en inglés y los estudios realizados en Brasil la han utilizado sin una traducción y análisis adecuados de propiedades psicométricas. Objetivo. Traducir y adaptar culturalmente la FAC para el portugués brasileño y probar su fiabilidad y validez concurrente en individuos afectados por ictus. Pacientes y métodos. La traducción y la adaptación transcultural implicaron seis pasos. La fiabilidad interevaluador se probó con cinco fisioterapeutas que vieron vídeos de la marcha de las personas afectadas por ictus y, después de un intervalo mínimo de una semana, los vieron por segunda vez para determinar la fiabilidad intraevaluador. La validez concurrente se determinó correlacionando la FAC con el resultado de la prueba de marcha de 10 metros (PM10m). Resultados. La fiabilidad intraevaluador varió entre un valor kappa de 0,68-0,95 (p = 0,001), y la fiabilidad interevaluador, un valor kappa de 0,517-0,794 (p = 0,001). La correlación entre la FAC y la PM10m fue rs = 0,771 (p = 0,001). Conclusión. La traducción, la adaptación transcultural y el análisis de las propiedades psicométricas demostraron que la FAC es una medida clínica válida y fiable para clasificar la deambulación de los individuos brasileños afectados por ictus en un entorno clínico.

Validation and reproducibility of the Glittre activities of daily living test for individuals with Parkinson's disease. / Validación y reproducibilidad de la prueba Glittre de actividades de la vida diaria en personas con enfermedad de Parkinson.

AIM: To investigate the validity and reproducibility of the Glittre Activities of Daily Living (Glittre-ADL) test for individuals with Parkinson's disease. SUBJECTS AND METHODS: Thirty individuals with Parkinson's disease and 19 healthy individuals (control group) were evaluated. Parkinson's disease group was evaluated by the Unified Parkinson's Disease Rating Scale (UPDRS) and underwent the Glittre-ADL test, six-minute walk test (6MWT) and ten-meter walk test (10mWT). Control group performed the Glittre-ADL test. For the intraobserver analysis, two Glittre-ADL tests were performed. For the interobserver analysis, the Glittre-ADL test was repeated on a different day by a second examiner. RESULTS: The Glittre-ADL test was significantly correlated with UPDRS Section II, Section III, and total score. The Glittre-ADL test was inversely correlated with the 6MWT and positively correlated with the 10mWT. The time required to perform the Glittre-ADL test was shorter on the retest in the intraobserver analysis and in the interobserver analysis. The mean difference between the first and second tests, the standard error of measurement and minimum detectable change in minutes were 0.40, 0.08 and 0.24, respectively, for intraobserver, and 0.40, 0.22 and 0.62, for interobserver. CONCLUSION: The Glittre-ADL test is valid and reproducible to evaluate functional capacity in individuals with Parkinson's disease.

TITLE: Validación y reproducibilidad de la prueba Glittre de actividades de la vida diaria en personas con enfermedad de Parkinson.Objetivo. Investigar la validez y la reproducibilidad de la prueba Glittre de actividades de la vida diaria (AVD-Glittre) para personas con enfermedad de Parkinson. Sujetos y métodos. Se evaluó a 30 pacientes con enfermedad de Parkinson y 19 sujetos sanos (grupo de control). El grupo con enfermedad de Parkinson fue evaluado con la Unified Parkinson's Disease Rating Scale (UPDRS) y sometido a la prueba AVD-Glittre, la prueba de marcha de seis minutos (6MWT) y la prueba de marcha de 10 metros (10mWT). El grupo de control realizó la prueba AVD-Glittre. Para el análisis intraobservador se realizaron dos pruebas AVD-Glittre, y para el análisis interobservador, la prueba se repitió otro día con un segundo examinador. Resultados. La prueba AVD-Glittre se correlacionó significativamente con la sección II, la sección III y la puntuación total de la UPDRS. Se correlacionó inversamente con la 6MWT y positivamente con la 10mWT. El tiempo requerido para realizar la prueba AVD-Glittre fue más corto en la nueva prueba en el análisis intraobservador y en el análisis interobservador. La diferencia de medias entre la primera y la segunda pruebas, el error estándar de medición y el cambio mínimo detectable en minutos fueron 0,40, 0,08 y 0,24, respectivamente, para el análisis intraobservador, y 0,40, 0,22 y 0,62, respectivamente, para el análisis interobservador. Conclusión. La prueba AVD-Glittre es válida y reproducible para evaluar la capacidad funcional en personas con enfermedad de Parkinson.

Elevada frequência de Aeromonas spp. e genes de virulência em cultivos de tilápia-do-nilo (Oreochromis niloticus) em tanques-rede, na região semiárida de Pernambuco, Brasil / High frequency of Aeromonas spp. and virulence genes in Nile Tilapia farms (Oreochromis niloticus) in semi-arid regions of Pernambuco, Brazil

Objetivou-se avaliar a ocorrência de Aeromonas spp. em peixes e amostras de água na região semiárida de Pernambuco e avaliar a frequência de aerolissina (aerA), enterotoxina citotóxica (act), enterotoxina citotônica (alt) e serina protease (ahp) nesses isolados. Foram analisados 70 peixes vivos e oito mortos com sinais clínicos de aeromoniose e 16 amostras de água. Aeromonas spp. foram identificadas por análises microbiológicas (provas bioquímicas) e molecular, usando-se primers específicos para a região 16S rRNA, e a distribuição dos quatro fatores de virulência (aerA, alt, act e ahp) foi investigada por ensaio de PCR. Cento e cinquenta e cinco (84,7%) isolados foram confirmados como Aeromonas spp. na análise molecular. Os genes de virulência mais frequentes foram act (53,55%) e aerA (51,61%). De acordo com o tipo de amostra, observou-se maior frequência do gene aerA (87,5% P=0,0474) em isolados de peixes mortos e a menor frequência do gene act (47,73% P=0,0002) em peixes vivos. Este estudo demonstrou a presença de aeromoniose no cultivo de tilápias em tanques-rede, nos municípios de Jatobá e Petrolândia, na região semiárida de Pernambuco. A detecção de aerA, act e alt pode ser utilizada na tipagem de virulência de Aeromonas spp.(AU)

The purpose of this study was to evaluate the occurrence of Aeromonas spp. from fishes and tilapia net-cage farm water in semi-arid regions of Pernambuco and to evaluate the frequency of the aerolysin (aerA), cytotoxic enterotoxin (act), cytotonic enterotoxin (alt) and serine protease (ahp) genes in Aeromonas isolates. 70 live and eight dead fish with aeromoniosis clinical signs and 16 water samples were analyzed. Aeromonas spp. isolated were identified by microbiological (biochemical evidence) and molecular analysis using specific primers for 16SrRNA region, while the distribution of four virulence factors, including aerA, alt, act and ahp, was investigated by PCR assay. One hundred fifty-five (84.7%) isolates were confirmed as Aeromonas spp. by molecular analysis. The most frequent virulence genes in isolates were act (53.55%) and aerA (51,61%). According to the kind of sample, the higher frequency of aerA gene (87.5% P= 0.0474) was observed in isolates from dead fish and the lowest frequency of act gene (47.73% P= 0.0002) from live fish. This study found the presence of aeromoniosis on tilapia farming in net-cages on Jatobá and Petrolândia counties in the semiarid Pernambuco region. The detection of aerA, act and alt can be used for virulence typing of Aeromonas spp. isolates.(AU)

The role of formulation and follicular pathway in voriconazole cutaneous delivery from liposomes and nanostructured lipid carriers.

In general, colloids provide increased cutaneous permeation of drugs. Still, skin interaction and main pathways for drug diffusion may vary depending on system and formulation characteristics. The knowledge of how different colloidal systems interact with biological membranes and the formulation impact on delivery is especially relevant for drugs that can be encapsulated in multiple nanosystems, as voriconazole (VOR). In here, we compared VOR release and permeation profile from liposomes (LP) and nanostructured lipid carriers (NLC) in aqueous colloidal dispersions and in gel formulations. Despite the controlled drug release provided by gel formulations, formulation only had a significant impact on drug skin accumulation from LP. The reduced mobility in gel formulations compromised follicle deposition and drug retention in the skin. Such a hypothesis was confirmed by permeation experiments evaluating follicle pathway influence. Follicular route also had an influence on delivery from NLC, which was only significant for total drug that reached the acceptor medium. These differences could be attributed to the mechanisms of colloid interaction with the skin and subsequent drug release. Follicle LP deposition and slow drug release leads to higher cutaneous amounts whilst NLC interaction with skin and fast drug release leads to fast drug diffusion and deeper penetration. By the low MIC50 values encountered against Trichophyton rubrum (∼ 0.001 µg/mL), permeated amounts could inhibit fungal growth, regardless the system. In conclusion, both LP and NLC seem to be valuable systems for cutaneous VOR delivery. Fluidic formulations could provide better efficiency for cutaneous drug delivery from LP.

Description of Larval Instars To Fill a Gap in Forensic Entomology: The Larvae of Paralucilia pseudolyrcea (Diptera: Calliphoridae).

A fundamental assumption of forensic entomology for estimating the postmortem interval is that insect species are accurately identified, which depends on diagnostic morphological characters. Larvae of the blow fly Paralucilia pseudolyrcea (Mello, 1969) (Diptera: Calliphoridae) were sampled from four corpses in the state of Paraná, Brazil, but despite the forensic importance of this species, morphological data for the identification of its larval instars are lacking, limiting its usefulness in such cases. Thus, the main goal of this study was to describe the larval instars of P. pseudolyrcea. The material was obtained from a colony established by larvae collected from a corpse of a murder case. Overall, the distribution of spines is a key character for identifying this species in the first, second and third instars. Other characteristics, such as the presence of an accessory oral sclerite, the small cirri, the number of lobes of the anterior spiracle and the morphology of posterior spiracles, separates P. pseudolyrcea from other necrophagous blow flies. The detailed morphological description provided here facilitates the identification of larval instars of P. pseudolyrcea and their differentiation from those of other calliphorid species.

Hepatic fibropoiesis in dogs naturally infected with Leishmania (Leishmania) infantum treated with liposome-encapsulated meglumine antimoniate and allopurinol.

Hepatic fibropoiesis in canine visceral leishmaniasis (CVL) were evaluated by histological (morphometrical collagen deposition) and immunohistochemical assays characterizing alpha-actin (α-SMA), vimentin, calprotectin (L1 antigen), and TGF-ß in 46 naturally infected dogs with Leishmania infantum treated with liposome-encapsulated meglumine antimoniate and allopurinol separately and in combination. Six treatment groups were defined: meglumine antimoniate encapsulated in nanometric liposomes (LMA), allopurinol (ALLOP); liposome-encapsulated meglumine antomoniate combined with allopurinol (LMA+ALLOP); empty liposomes (LEMP); empty liposomes combined with allopurinol (LEMP+ALLOP) and saline. Relative liver weight was lower in LMA, LMA+ALLOP, and ALLOP groups compared to the LEMP control. Significantly lower granulomatous chronic inflammatory reaction was seen in the ALLOP group compared to a control group. Calprotectin was lowest in liver of those dogs showing lower numbers of intralobular hepatic granulomas. Collagen deposits were significantly higher in LMA compared to ALLOP, LEMP+ALLOP, and Saline groups. LMA+ALLOP group collagen deposition was higher than dogs treated only with allopurinol. Immunohistochemical analysis showed significant higher α-SMA in hepatic stellate cells (HSCs), hepatic perisinusoidal cells, in control groups than LMA+ALLOP and LEMP+ALLOP. Alpha-actin and Vimentin positive cells were diffusely distributed throughout the liver parenchyma in the hepatic lobule, mainly in HSCs. Vimentin expression was significantly higher in the saline group than in the ALLOP group. Our data suggest that allopurinol inhibits HSC and results in lower collagen deposits in liver during CVL progression, as supported by the significantly lower expression of TGF-ß in the ALLOP group compared to other groups. Results demonstrated that treatment with allopurinol inhibited chronic granulomatous inflammatory reaction and hepatic fibrosis in CVL.

Toxin Gene Contents and Activity of Bacillus thuringiensis Strains Against Two Sugarcane Borer Species, Diatraea saccharalis (F.) and D. flavipennella (Box).

Bacillus thuringiensis (Berliner) bears essential characteristics in the control of insect pests, such as its unique mode of action, which confers specificity and selectivity. This study assessed cry gene contents from Bt strains and their entomotoxicity against Diatraea saccharalis (F.) and Diatraea flavipennella (Box) (Lepidoptera: Crambidae). Bioassays with Bt strains were performed against neonates to evaluate their lethal and sublethal activities and were further analyzed by PCR, using primers to identify toxin genes. For D. saccharalis and D. flavipennella, 16 and 18 strains showed over 30% larval mortality in the 7th day, respectively. The LC50 values of strains for D. saccharalis varied from 0.08 × 105 (LIIT-0105) to 4104 × 105 (LIIT-2707) spores + crystals mL-1. For D. flavipennella, the LC50 values of strains varied from 0.40 × 105 (LIIT-2707) to 542 × 105 (LIIT-2109) spores + crystals mL-1. For the LIIT-0105 strain, which was the most toxic to D. saccharalis, the genes cry1Aa, cry1Ab, cry1Ac, cry1B, cry1C, cry1D, cry1F, cry1I, cry2Aa, cry2Ab, cry8, and cry9C were detected, whereas for the strain LIIT-2707, which was the most toxic to D. flavipennella, detected genes were cry1Aa, cry1Ab, cry1Ac, cry1B, cry1D, cry1F, cry1I, cry2Aa, cry2Ab, and cry9. The toxicity data and toxin gene content in these strains of Bt suggest a great variability of activity with potential to be used in the development of novel biopesticides or as source of resistance genes that can be expressed in plants to control pests.

Evaluation of microbial qPCR workflows using engineered Saccharomyces cerevisiae.

AIMS: We describe the development and interlaboratory study of modified Saccharomyces cerevisiae as a candidate material to evaluate a full detection workflow including DNA extraction and quantitative polymerase chain reaction (qPCR). METHODS AND RESULTS: S. cerevisiae NE095 was prepared by stable insertion of DNA sequence External RNA Control Consortium-00095 into S. cerevisiae BY4739 to convey selectivity. For the interlaboratory study, a binomial regression model was used to select three cell concentrations, high (4 × 10(7) cells ml(-1)), intermediate (4 × 10(5) cells ml(-1)) and low (4 × 10(3) cells ml(-1)), and the number of samples per concentration. Seven participants, including potential end users, had combined rates of positive qPCR detection (quantification cycle <37) of 100%, 40%, and 0% for high, intermediate, and low concentrations, respectively. CONCLUSIONS: The NE095 strain was successfully detected by all participants, with the high concentration indicating a potential target concentration for a reference material. SIGNIFICANCE AND IMPACT OF THE STUDY: The engineered yeast has potential to support measurement assurance for the analytical process of qPCR, encompassing the method, equipment, and operator, to increase confidence in results and better inform decision-making in areas of applied microbiology. This material can also support process assessment for other DNA-based detection technologies.

Changes in postural control in patients with Parkinson's disease: a posturographic study.

OBJECTIVES: Postural instability is one of the most disabling features in Parkinson's disease (PD), and often leads to falls that reduce mobility and functional capacity. The objectives of this study were to analyse the limit of stability (LOS) and influence of the manipulation of visual, somatosensorial and visual-vestibular information on postural control in patients with PD and healthy subjects. DESIGN: Cross-sectional. SETTING: Movement Disorders Unit, university setting. PARTICIPANTS: Eighty-two subjects aged between 37 and 83 years: 41 with Parkinson's disease in the 'on' state and 41 healthy subjects with no neurological disorders. Both groups were matched in terms of sex and age. MAIN OUTCOME MEASURES: Unified Parkinson's Disease Rating Scale (UPDRS)-motor score, modified Hoehn and Yahr staging, Dynamic Gait Index (DGI) and posturography with integrated virtual reality. The parameters analysed by posturography were LOS area, area of body centre of pressure excursion and balance functional reserve in the standing position in 10 conditions (open and closed eyes, unstable surface with eyes closed, saccadic and optokinetic stimuli, and visual-vestibular interaction). RESULTS: The mean UPDRS motor score and DGI score were 27 [standard deviation (SD) 14] and 21 (SD 3), respectively. Thirteen participants scored between 0 and 19 points, indicating major risk of falls. Posturographic assessment showed that patients with PD had significantly lower LOS area and balance functional reserve values, and greater body sway area in all posturographic conditions compared with healthy subjects. CONCLUSIONS: Patients with PD have reduced LOS area and greater postural sway compared with healthy subjects. The deterioration in postural control was significantly associated with major risk of falls.

Toxicidade subaguda do extrato etanólico das folhas de Myracrodruon urundeuva sobre o ciclo estral de ratas Wistar / Subacute toxicity of Myracrodruon urundeuva leave ethanolic extract and its effects on the estrous cycle of female Wistar rats

RESUMO A fitoterapia, abrangendo o uso popular e empírico de plantas medicinais no combate e prevenção de doenças, deve ser aliada a estudos científicos que comprovem a eficácia e segurança desses compostos. No Brasil, a ampla biodiversidade vegetal da região nordeste favorece o uso e estudo de plantas com potenciais terapêuticos. A Aroeira do Sertão (Myracrodruon urundeuva Allem.) é uma espécie pertencente à família Anacardiaceae, comum no semiárido, desde o Piauí até Minas Gerais. Seu uso é diverso e disseminado por todo o País, sendo indispensáveis estudos de suas potencialidades e riscos. Devido à ampla utilização da Myracrodruon Urundeuva de forma empírica, este trabalho tem por objetivo a pesquisa de efeitos tóxicos com doses repetidas do extrato etanólico de Myracrodruon urundeuva Allem. (EEMU) e sua influência no ciclo estral de ratas Wistar. No protocolo de toxicidade subaguda foram utilizadas 25 ratas Wistar, divididas em 5 grupos (n=5), tratadas com diferentes doses do EEMU (125, 250, 500 e 1000 mg/kg) e água destilada (controle) por um período de 28 dias. Nesse período foram realizadas mensurações do consumo de água e ração e avaliação comportamental. Esses animais foram avaliados diariamente quanto a fase do ciclo estral, por meio de esfregaço vaginal a fresco, sendo observada a frequência de cada fase bem como o intervalo interestro. As ratas foram anestesiadas e o sangue foi coletado para a realização dos ensaios bioquímicos. Em seguida, foram eutanasiadas para coleta e avaliação dos órgãos internos. Durante o período de avaliação, não foram observadas alterações de comportamento, nem de consumo de água ou ração. A evolução ponderal dos animais não diferiu entre os grupos tratados. A concentração sérica de ALT foi maior nos animais tratados com EEMU 1000 mg/kg. As ratas tratadas com o extrato não apresentaram alterações significativas na frequência das fases do ciclo estral bem como duração do estro e intervalo entre estros, quando comparadas ao grupo controle. Os resultados obtidos neste estudo não apontam alterações tóxicas significativas, sistêmicas ou sobre o ciclo estral, de ratas Wistar tratadas com o extrato, nas doses avaliadas, por um período de 28 dias.

ABSTRACT The herbal medicine, comprising the popular and empirical use of medicinal plants in combating and preventing diseases, should be combined with scientific studies that prove the effectiveness and safety of these compounds. In Brazil, the vast plant biodiversity of the Northeast favors the use and study of plants with therapeutic potential. Aroeira (Myracrodruon urundeuva Allem.) is a species of the Anacardiaceae family, common in the semiarid region, from Piauí to Minas Gerais. Its use is diverse and widespread throughout the country, making studies of its potential and risks necessary. Due to the wide use of Myracrodruon urundeuva in an empirical form, this paper aims to research toxic effects with repeated doses of Myracrodruon urundeuva Allem.ethanol extract (EEMU) and its influence on the estrous cycle of female Wistar rats. In a subacute toxicity protocol 35 female Wistar rats were used, divided into 5 groups (n=5) treated with different doses of EEMU (125, 250, 500 and 1000 mg/kg) and distilled water (control) for 28 days. In this period measurements of water intake, feed consumption were carried out and a behavioral assessment was performed. The estrous cycles of these animals were evaluated daily by fresh vaginal smear, it the frequency of each phase as well as the inter-estrus intervals were observed. The rats were anesthetized and blood was collected for the biochemical assays. Then they were euthanized for the collection and evaluation of internal organs. During the evaluation period, there were no observed behavioral changes nor were the water or food consumption variations. The weight gain of the animals did not differ between treatment groups. The serum ALT was higher in animals treated with EEMU 1000 mg/kg. The rats treated with the extract showed no significant changes in the frequency of the phases of the estrous cycle, estrus duration and estrus interval when compared to the control group. The results of this study do not indicate significant toxic alterations, neither systemic nor on the estrous cycle in female rats treated with the extract, at the evaluated doses, over a period of 28 days.

A high efficient protocol for soybean root transformation by Agrobacterium rhizogenes and most stable reference genes for RT-qPCR analysis.

KEY MESSAGE: A 55% transformation efficiency was obtained by our optimized protocol; and we showed that GmELF1 - ß and GmELF1 - α are the most stable reference genes for expression analyses under this specific condition. Gene functional analyses are essential to the validation of results obtained from in silico and/or gene-prospecting studies. Genetic transformation methods that yield tissues of transient expression quickly have been of considerable interest to researchers. Agrobacterium rhizogenes-mediated transformation methods, which are employed to generate plants with transformed roots, have proven useful for the study of stress caused by root phytopathogens via gene overexpression and/or silencing. While some protocols have been adapted to soybean plants, transformation efficiencies remain limited; thus, few viable plants are available for performing bioassays. Furthermore, mRNA analyses that employ reverse transcription quantitative polymerase chain reactions (RT-qPCR) require the use of reference genes with stable expression levels across different organs, development steps and treatments. In the present study, an A. rhizogenes-mediated soybean root transformation approach was optimized. The method delivers significantly higher transformation efficiency levels and rates of transformed plant recovery, thus enhancing studies of soybean abiotic conditions or interactions between phytopathogens, such as nematodes. A 55% transformation efficiency was obtained following the addition of an acclimation step that involves hydroponics and different selection processes. The present study also validated the reference genes GmELF1-ß and GmELF1-α as the most stable to be used in RT-qPCR analysis in composite plants, mainly under nematode infection.

Starch-based Pickering emulsions for topical drug delivery: A QbD approach.

Pickering emulsions are stabilized by solid particles instead of surfactants and have been widely investigated in pharmaceutical and cosmetic fields since they present less adverse effects than the classical emulsions. A quality by design (QbD) approach was applied to the production of w/o emulsions stabilized by starch. A screening design was conducted to identify the critical variables of the formula and the process affecting the critical quality properties of the emulsion (droplet size distribution). The optimization was made by establishing the Design Space, adjusting the concentration of starch and the quantity of the internal aqueous phase. The emulsion production process was, in turn, adjusted by varying the time and speed of stirring, to ensure quality and minimum variability. The stability was also investigated, demonstrating that an increase in starch concentration improves the stability of the emulsion. Rheological and mechanical studies indicated that the viscosity of the emulsions was enhanced by the addition of starch and, to a higher extent, by the presence of different lipids. The developed formulations was considered non-irritant, by an in vitro assay using human cells from skin (Df and HaCat) with the cell viability higher than 90% and, with self-preserving properties. Finally, the QbD approach successfully built quality in Pickering emulsions, allowing the development of hydrophilic drug-loaded emulsions stabilized by starch with desired organoleptic and structural characteristics. The results obtained suggest that these systems are a promising vehicle to be used in products for topical administration.

Structural and functional studies of Hsp70-escort protein--Hep1--of Leishmania braziliensis.

Hep1 is a mitochondrial Hsp70 (mtHsp70) co-chaperone that presents a zinc finger domain essential for its function. This co-chaperone acts to maintain mtHsp70 in its soluble and functional state. In this work, we have demonstrated that Leishmania braziliensis mtHsp70 (LbmtHsp70) is also dependent on the assistance of Hep1. To understand the L. braziliensis Hep1 (LbHep1) structure-function relationship, we produced LbHep1 and two truncated mutants corresponding to the C-terminal zinc finger domain and the N-terminal region. We observed that LbHep1 is composed of an unfolded N-terminal region and a ß-sheet-folded C-terminal domain, which holds the zinc-binding motif. Both LbHep1 and the zinc finger domain construction maintained LbmtHsp70 solubility in co-expression systems after cell lysis. In solution, LbHep1 behaved as a highly elongated monomer, probably due to the unfolded N-terminal region. Furthermore, we also observed that the zinc ion interacted with LbHep1 with high affinity and was critical for LbHep1 structure and stability because its removal from LbHep1 solutions altered the protein structure and stability. In vitro, LbHep1 protected, in sub-stoichiometric fashion, LbmtHsp70 from thermally induced aggregation but did not present intrinsic chaperone activity on model client proteins. Therefore, LbHep1 is a specific chaperone for LbmtHsp70.