[Effects of coffee consumption on salivary cortisol and alpha amylase in young adults]. / Efectos del consumo de café sobre el cortisol y la alfa-amilasa salival en adultos jóvenes.

Objective: The purpose of this study was to determine the effects of coffee consumption on salivary cortisol (sCort) and alpha amylase (sAA) in young adults. Materials and methods: Sixty healthy university students, habitual coffee consumers, participated in this descriptive observational study. Participants were divided into three groups: G1 low consumption (≤ 2 cups of coffee per day, n = 20), G2 moderate consumption (2-5 cups of coffee per day, n = 20), and G3 high consumption (>5 cups of coffee per day, n = 20). Saliva self-collection was in the morning (6:30-7:30 AM) and at night (08:00-09:00 PM). sCort was analyzed using chemiluminescence and sAA activity by kinetic method. Statistical analysis of the data was performed using Student's t-test and analysis of variance. Results: The sample consisted of 30 women and 30 men, aged between 20 and 35 years. In all groups, sCort values were higher in the morning (AM 0,29 ± 0,19 vs. PM 0,09 ± 0,05 µg/dl, p < 0.0001). In contrast, sAA levels were higher in the night (PM 160,16 ± 60,42 vs. AM 32,79 ± 12,98 U/ml, p < 0.0001). No significant differences were detected, in the contents of Corts and AAs, between the groups. Conclusion: : Coffee consumption, in non-stressful conditions, did not alter levels and patterns of sCort and sAA in young adults.

Salivary alpha-amylase stress reactivity in advanced-aged marmosets (Callithrix jacchus): Impacts of cognitive function and oral health status.

Salivary alpha-amylase (sAA) is an enzyme found in saliva and is considered a noninvasive biomarker for sympathetic nervous system activity. While a wide range of sAA activity in response to stress has been reported in nonhuman primates, the effects of stress on sAA activity in common marmosets are still unknown. We tested the hypothesis that advanced age and cognitive function may have an impact on stress-related sAA reactivity in marmosets. Thirteen marmosets (nine males and five females) had saliva samples collected during a stressful condition (manual restraint stress) at two different time points, with a 60-min interval. On the next day, the animals underwent the object recognition test (ORT, a type of cognitive test), and then oral examinations. The animals were categorized into two age groups: old (10-13 years), and very old (15-22 years). Irrespective of age, sAA levels showed a significant difference between T1 (mean 2.07 ± 0.86 U/mL) and T2 samples (mean 1.03 ± 0.67 U/mL), with higher values observed at T1 (p < 0.001). The intra-assay coefficients of variation (CV) for low and high sAA concentrations were 10.79% and 8.17%, respectively, while the interassay CVs for low and high sAA concentrations were 6.39% and 4.38%, respectively. Oral health issues were common but did not significantly impact sAA levels. The ORT indicated that the animals could recognize an object placed in the cage 6 h after familiarization. In conclusion, all marmosets showed a higher sAA concentration in the first saliva sample as compared to the second saliva sample collected 1 h later, indicating adaptation to stress. No significant differences in sAA levels were observed between sexes, ORT performance, or oral health. Our results indicate that autonomic responsivity and cognitive (memory) functions were preserved even in very old marmosets.

Evaluation of inhibitory activity, purification and X-ray crystallography of Alpha-Amylase inhibitor from Phaseolus vulgaris cultivars of Uttarakhand, India / Avaliação da atividade inibitória, purificação e cristalografia de raios X do inibidor Alfa-Amilase de cultivares de Phaseolus vulgaris de Uttarakhand, Índia

Abstract The present work is based on analysis of inhibitory activity of alpha-amylase inhibitor in selected cultivars of Phaseolus vulgaris of Uttarakhand. Fifteen samples were assessed for inhibitory activity of alpha-amylase inhibitor. Significant variations were found in different cultivars. Crude extract of alpha-amylase inhibitor from sample PUR (Purola) have shown maximum inhibitory activity (70.2 ± 0.84). Crude extract of all the cultivars have shown considerable variations in inhibitory activity in the temperature ranging from 20ºC to 100ºC. Based on inhibitory activity and heat stability profile, the alpha amylase inhibitor was purified from PUR cultivar. The purified inhibitor was found to be stable even at 90ºC with an inhibitory activity of 97.20 ±0.09. The molecular weight of purified inhibitor on Native PAGE (Polyacrylamide gel electrophoresis) was found to be 31kd, consisting of two subunits of 17kd and 14kd on SDS-PAGE.

O presente trabalho é fundamentado na análise da atividade inibitória do inibidor da alfa-amilase em cultivares selecionadas de Phaseolus vulgaris, de Uttarakhand. Quinze amostras foram avaliadas quanto à atividade inibitória do inibidor da alfa-amilase. Variações significativas foram encontradas em diferentes cultivares. O extrato bruto do inibidor da alfa-amilase da amostra PUR (Purola) apresentou atividade inibitória máxima (70,2 ± 0,84). O extrato bruto de todas as cultivares apresentou variações consideráveis ​​na atividade inibitória na temperatura de 20ºC a 100ºC. Com base na atividade inibitória e no perfil de estabilidade ao calor, o inibidor da alfa-amilase foi purificado do cultivar PUR. O inibidor purificado mostrou-se estável mesmo a 90ºC, com uma atividade inibitória de 97,20 ± 0,09. O peso molecular do inibidor purificado em Native PAGE (eletroforese em gel de poliacrilamida) foi de 31kd, consistindo em duas subunidades de 17kd e 14kd em SDS-PAGE.

Aspergillus clavatus UEM 04: An efficient producer of glucoamylase and α-amylase able to hydrolyze gelatinized and raw starch.

The best amylolytic activity production by Aspergillus clavatus UEM 04 occurred in submersed culture, with starch, for 72 h, at 25 °C, and 100 rpm. Exclusion chromatography partially purified two enzymes, which ran as unique bands in SDS-PAGE with approximately 84 kDa. LC-MS/MS identified a glucoamylase (GH15) and an α-amylase (GH13_1) as the predominant proteins and other co-purified proteins. Zn2+, Cu2+, and Mn2+ activated the glucoamylase, and SDS, Zn2+, Fe3+, and Cu2+ inhibited the α-amylase. The α-amylase optimum pH was 6.5. The optimal temperatures for the glucoamylase and α-amylase were 50 °C and 40 °C, and the Tm was 53.1 °C and 56.3 °C, respectively. Both enzymes remained almost fully active for 28-32 h at 40 °C, but the α-amylase thermal stability was calcium-dependent. Furthermore, the glucoamylase and α-amylase KM for starch were 2.95 and 1.0 mg/mL, respectively. Still, the Vmax was 0.28 µmol/min of released glucose for glucoamylase and 0.1 mg/min of consumed starch for α-amylase. Moreover, the glucoamylase showed greater affinity for amylopectin and α-amylase for maltodextrin. Additionally, both enzymes efficiently degraded raw starch. At last, glucose was the main product of glucoamylase, and α-amylase produced mainly maltose from gelatinized soluble starch hydrolysis.

"Do it yourself" protocol to fabricate dual-detection paper-based analytical device for salivary biomarker analysis.

This paper describes the design and construction of dual microfluidic paper-based analytical devices (dual-µPADs) as a lab-on-paper platform involving a "do-it-yourself" fabrication protocol. The device comprises a colorimetric and electrochemical module to obtain a dual-mode signal readout sensing strategy. A 3D pen polymeric resin was used to prepare graphite carbon-based electrodes and hydrophobic barriers on paper substrates. The proposed carbon-based ink was employed to manufacture electrodes on paper based on a stencil-printing approach, which were further characterized by electrochemical and morphological analyses. The analytical performance of the dual-µPADs was simultaneously evaluated for lactate, pH, nitrite, and salivary amylase (sAA) analysis. To demonstrate the proof-of-concept, saliva samples collected from both healthy individuals and those with periodontitis were successfully tested to demonstrate the feasibility of the proposed devices. Samples collected from individuals previously diagnosed with periodontitis showed high levels of nitrite and sAA (> 94 µmol L-1 and > 610 U mL-1) in comparison with healthy individuals (≤ 16 µmol L-1 and 545 U mL-1). Moreover, periodontitis saliva resulted in acid solution and almost null lactate levels. Notably, this protocol supplies a simple way to manufacture dual-µPADs, a versatile platform for sensitive detecting of biomarkers in saliva playing a crucial role towards the point-of-care diagnosis of periodontal disease.

Ewes fed high-concentrate diets containing flint corn and increasing levels of exogenous amylolytic enzyme: effects on nutrient intake and digestibility / Ovelhas alimentadas com dietas de alto teor de concentrado contendo milho flint e níveis crescentes de enzima amilolítica exógena: efeitos sobre o consumo e a digestibilidade dos nutrientes

The aim was to evaluate if the inclusion of exogenous amylolytic enzyme affect the nutrient intake and digestibility in ewes fed high-concentrate diets containing flint corn. Five Santa Inês × Dorper crossbred ewes (54.04 ± 4.5 kg and aged 8 months) were used in a 5 x 5 Latin square design. All animals were housed in individual metabolic cages for 60 days. The treatments consisted of a control diet (without amylolytic enzyme) and four inclusion levels of an amylolytic enzyme (3,000, 6,000, 9,000, and 12,000 α-amylase dextrinizing units [DU] kg-1 dry matter [DM]). The enzyme was mixed into the feed at the time of supply to the animals. Data were analyzed by ANOVA, and orthogonal polynomial contrasts were used. Nutrient intake was not influenced by amylolytic enzyme inclusion. The digestibility of DM, organic matter, neutral detergent fiber, total carbohydrates, non-fibrous carbohydrates, and gross energy showed a quadratic increase with enzyme inclusion (P<0.05), with maximum values at levels of 7,600, 7,500, 6,300, 7,500, 7,400, and 7,800 DU kg-1 DM, respectively. Total digestible nutrients of diets also showed a quadratic increase, with a maximum value of 894 g kg-1 at a level of α-amylase activity of 7,786 DU kg-1 DM. The inclusion of the exogenous amylolytic enzyme from 6,300 to 7,800 DU kg-1 DM doesn't alter nutrient intake and improves the digestibility in ewes fed high-concentrate diets.(AU)

Objetivou-se avaliar se a inclusão de enzima amilolítica exógena afeta a ingestão e a digestibilidade dos nutrientes em ovelhas alimentadas com dietas de alto teor de concentrado contendo milho flint. Foram usadas cinco ovelhas mestiças Santa Inês × Dorper (54,04 ± 4,5 kg e 8 meses de idade) em um quadrado latino 5x5. Todos os animais foram alojados em gaiolas metabólicas individuais por 60 dias. Os tratamentos consistiram em uma dieta controle (sem enzima amilolítica) e quatro níveis de enzima amilolítica (3.000; 6.000; 9.000 e 12.000 unidades de dextrinização [UD] de α-amilase kg-1 de matéria seca [MS]). A enzima foi misturada com o alimento no momento do fornecimento aos animais. Os dados foram analisados por ANOVA e contrates ortogonais polinomiais. O consumo de nutrientes não foi influenciado pela inclusão da enzima amilolítica. A digestibilidade da MS, matéria orgânica, fibra em detergente neutro, carboidratos totais, carboidratos não fibrosos e energia bruta apresentaram efeito quadrático com a inclusão de enzima (P<0,05), com valores máximos nos níveis de inclusão da enzima 7.600; 7.500; 6.300; 7.500; 7.400 e 7.800 UD kg-1 MS, respectivamente. Os nutrientes digestíveis totais também apresentaram efeito quadrático, com máximo valor de 894 g kg-1 no nível de atividade de α-amilase de 7.786 UD kg-1 MS. A inclusão da enzima amilolítica exógena entre 6.300 e 7.800 UD kg-1 MS não modifica o consumo de nutrientes e melhora a digestibilidade de ovelhas alimentadas com dietas de alto teor de concentrado.(AU)

Bioactivity evaluation of least explored traditionally acclaimed medicinally potent herb Nanorrhinum ramosissimum(Wall. ) Betsche

Abstract Present study analysed the therapeutic potential of traditionally acclaimed medicinal herb Nanorrhinum ramosissimum, using plant parts extracted with different solvents (10 mg/mL). Shoot extracts exhibited comparatively better antimicrobial properties, in comparison to root extracts. Total phenolic content was estimated, to ascertain its dependency on antioxidant properties of plant extracts. Antioxidant assay revealed promising results in comparison to IC50 value of standard ascorbic acid (52.2±0.07 µg/mL), for methanolic extracts of shoot (61.07±0.53 µg/mL and 64.33±0.33 µg/mL) and root (76.705±0.12 µg/mL and 89.73±0.28 µg/ mL) for in vivo and in vitro regenerants respectively. Correlation coefficient R2 values ranged between 0.90-0.95, indicating a positive correlation between phenolic contents and antioxidant activity. Plant extracts were also able to inhibit DNA oxidative damage again indicating their antioxidative potential. Antidiabetic potential was confirmed by alpha amylase inhibition assay where shoot methanolic extracts (invivo, in vitro) exhibited the best IC50 values (54.42±0.16 µg/mL, 66.09±0.12 µg/mL) in comparison to standard metformin (41.92±0.08 µg/mL). Ethanolic extracts of roots (in vitro, invivo) exhibited the relative IC50 values (88.97±0.32µg/mL,96.63±0.44 µg/mL) indicating that shoot parts had a better alpha amylase inhibition property; thus proving the herb's bioactive potential and its prospective therapeutic source for curing various ailments.

Extracts of Fruits and Plants Cultivated In Vitro of Aristotelia chilensis (Mol.) Stuntz Show Inhibitory Activity of Aldose Reductase and Pancreatic Alpha-Amylase Enzymes.

Aristotelia chilensis is a plant whose fruit is considered a powerful natural antioxidant. During the last years, some investigations of the fruit have been carried out, finding antioxidant properties in the juice or the phenolic fraction. The antioxidant properties of the plant are useful in the inhibition of enzymes related to diabetes such as pancreatic aldose reductase and alpha-amylase. Because many synthetic drugs used today have limitations and potentially harmful side effects, the use of naturally occurring compounds, such as flavonoids, is clinically attractive. In this study, the characterization of aqueous extracts of fruits and in vitro plants of A. chilensis was carried out based on their content of anthocyanins and total phenols, the antioxidant capacity by the antiradical activity 2,2-diphenyl-1-picrilhydrazil (DPPH), and the profile of anthocyanins and other phenolic compounds by liquid chromatography coupled to mass spectrometry (LC-MS/MS). Subsequently, the effect of these extracts on the inhibition of bovine aldose reductase and pancreatic alpha-amylase enzymes was determined. According to our results, extracts of fruits and in vitro plants of A. chilensis achieved inhibition of the bovine aldose reductase enzyme of 85.54 ± 1.86% and 75.67 ± 1.21%, respectively. Likewise, the percentage of inhibition of the pancreatic alpha-amylase enzyme for fruit extracts was 29.64 ± 0.63%, while for in vitro plant extracts it was 47.66 ± 0.66%. The antioxidant and enzymatic inhibition activity of the extracts were related to the content of anthocyanins, such as delphinidin and cyanidin glycosides as well as the phenols derived from quercetin, myricetin, and kaempferol. The results obtained allow us to suggest that the in vitro culture of plants of A. chilensis represents a viable biotechnological alternative to obtain phenolic compounds for the inhibition of aldose reductase and pancreatic alpha-amylase enzymes.

Debonding of orthodontic appliance changes salivary physicochemical properties and favors regression of active caries lesions: A 13-week follow-up study.

AIM: This prospective cohort study aimed to investigate whether orthodontic appliance removal (OAR) combined with caries-preventive strategies and fluoride varnish treatments alters salivary physicochemical properties, changes the activity of carbonic anhydrase VI (CA VIACT ) and α-amylase (α-AMLACT ), and favors the regression of active caries lesions (ACL). DESIGN: Twenty-two individuals aged between 13 and 24 years were assessed for the presence of visible biofilm, daily sugar exposure, caries activity, salivary flow rate (SFR), pH, buffering capacity (BC), and CA VIACT and α-AMLACT activity at baseline, and 1, 5, and 13 weeks after OAR. Variables were assessed using repeated-measures analysis of variance, Cochran's Q and McNemar's test, and Pearson's correlation. RESULTS: We observed a significant decrease in the number of ACL at the 5-week (29% reduction) and 13-week follow-ups (58% reduction). At the 5- and 13-week follow-ups, the percentage of visible biofilm and sugar exposure decreased, whereas the salivary pH and α-AMLACT activity significantly increased. BC and CA VIACT remained unchanged throughout the follow-up. CONCLUSION: OAR combined with caries-preventive strategies and fluoride varnish treatments favored the regression of ACL and increased salivary pH and α-AMLACT activity, whereas BC and CA VIACT remained stable.

Immunosensor Based on Zinc Oxide Nanocrystals Decorated with Copper for the Electrochemical Detection of Human Salivary Alpha-Amylase.

(1) Background: Nanocrystals (NCs)-based electrochemical sensors have been proposed for biomarkers detection, although immunosensors using ZnO NCs decorated with copper are still scarce. (2) Methods: Electrochemical immunodetection of human salivary alpha-amylase (HSA) used ZnO, CuO, and ZnO:xCu (x = 0.1, 0.4, 1.0, 4.0, and 12.0) NCs. (3) Results: Substitutional incorporation of Cu2+ in the crystalline structure of ZnO and formation of nanocomposite were demonstrated by characterization. Graphite electrodes were used and the electrochemical signal increased by 40% when using ZnO:1Cu and 4Cu (0.25 mg·mL-1), in an immunosensor (0.372 mg·mL-1 of anti-alpha-amylase and 1% of casein). Different interactions of HSA with the alpha-amylase antibody were registered when adding the NCs together, either before or after the addition of saliva (4 µL). The immunosensor changed specificity due to the interaction of copper. The ZnO:1Cu and ZnO:4Cu samples showed 50% interference in detection when used before the addition of saliva. The immunosensor showed 100% specificity and a sensitivity of 0.00196 U·mL-1. (4) Conclusions: Results showed that the order of NCs addition in the sensors should be tested and evaluated to avoid misinterpretation in detection and to enable advances in the validation of the immunosensor.

Biological properties and phytochemical characterization from Miconia chamissois Naudin aqueous extract / Propiedades biológicas y caracterización fitoquímica del extracto acuoso de Miconia chamissois Naudin

The objective of this study was to evaluate biological and phytochemical properties of the aqueous extract from the leaves of Miconia chamissois Naudin (AEMC). Phytochemical properties were assessed by analyzing the chromatographic profile and the polyphenol content of AEMC. Biological properties evaluation was conducted based on cytotoxicity assay and by evaluating the antioxidant, antimicrobial, and enzymatic inhibition activities. Results indicated the presence of phytochemicals in AEMC such as flavonoids and polyphenols, including rutin, isoquercitrin and vitexin derivatives. AEMC showed antioxidant activity, which may be attributed to the high polyphenolic content. Moreover, AEMC demonstrated in vitro enzyme inhibition activity against tyrosinase and alpha-amylase, as well as showed low cytotoxicity. On the other hand, AEMC exhibited weak antimicrobial activity against S. aureusand C. albicans. Thus, AEMC is a promising alternative in search of potential drugs for the treatment of diseases induced by oxidative stress and inflammation, conditions due to hyperpigmentation processes, such as melisma, as well as for diabetes.

El objetivo de este estudio fue detectar las propiedades biológicas y fitoquímicos del extracto acuoso de las hojas de Miconia chamissois Naudin (AEMC). Las propiedades fitoquímicas se evaluaron analizando el perfil cromatográfico y el contenido de polifenoles de AEMC. La evaluación de las propiedades biológicas se realizó en base al ensayo de citotoxicidad y evaluando las actividades de inhibición antioxidante, antimicrobiana y enzimática. Los resultados indicaron la presencia de fitoquímicos en AEMC, como flavonoides y polifenoles, que incluyen derivados de rutina, isoquercitrina y vitexina. AEMC mostró una actividad antioxidante considerable, que puede atribuirse al alto contenido polifenólico. Además, AEMC exhibió actividad de inhibición enzimática in vitro contra tirosinasa y alfa-amilasa, así como mostró baja citotoxicidad. Por otro lado, AEMC demostró actividad antimicrobiana débil contra S. aureusy C. albicans. Por lo tanto, AEMC es una alternativa prometedora en busca de posibles drogas para el tratamiento de enfermedades inducidas por el estrés oxidativo y la inflamación, afecciones debidas a procesos de hiperpigmentación, como el melasma, así como para la diabetes.

Modifying the catalytic preference of alpha-amylase toward n-alkanes for bioremediation purposes using in silico strategies.

Since the beginning of oil exploration, whole ecosystems have been affected by accidents and bad practices involving petroleum compounds. In this sense, bioremediation stands out as the cheapest and most eco-friendly alternatives to reverse the damage done in oil-impacted areas. However, more efforts must be made to engineer enzymes that could be used in the bioremediation process. Interestingly, a recent work described that α-amylase, one of the most evolutionary conserved enzymes, was able to promiscuously degrade n-alkanes, a class of molecules abundant in the petroleum admixture. Considering that α-amylase is expressed in almost all known organisms, and employed in numerous biotechnological processes, using it can be a great leap toward more efficient applications of enzyme or microorganism-consortia bioremediation approaches. In this work, we employed a strict computational approach to design new α-amylase mutants with potentially enhanced catalytic efficiency toward n-alkanes. Using in silico techniques, such as molecular docking, molecular dynamics, metadynamics, and residue-residue interaction networks, we generated mutants potentially more efficient for degrading n-alkanes, L183Y, and N314A. Our results indicate that the new mutants have an increased binding rate for tetradecane, the longest n-alkane previously tested, which can reside in the catalytic center for more extended periods. Additionally, molecular dynamics and network analysis showed that the new mutations have no negative impact on protein structure than the WT. Our results aid in solidifying this enzyme as one more tool in the petroleum bioremediation toolbox.

Change in malate dehydrogenase and alpha amylase activities in Rubus fruticosus and Valeriana jatamansi treated granary weevil, Sitophilus granarius / Mudança nas atividades de malato desidrogenase e alfa amilase em Rubus fruticosus e gorgulho tratado com Valeriana jatamansi, Sitophilus granarius

Poor storage conditions provide favorable environment to stored grain pests for their growth. The bio-pesticides are the best alternatives to synthetic pesticides. Present study was conducted to compare toxicity of Rubus fruticosus and Valeriana jatamansi against granary weevil, Sitophilus granarius and subsequent changes in enzyme activity responsible for grain damage. In current research 5 g of R. fruticosus fruit and V. jatamansi rhizome powders were tested separately against S. granarius, in 50 g wheat whole grains for seven days in comparison with the control. The enzymatic activity of malate dehydrogenase and alpha-amylase was observed in the cellular extracts of S. granarius. The insects were crushed and homogenized in phosphate-buffer solution and centrifuged at 10000 rpm for 5 minutes. For the enzymatic measurement supernatant was tested; the spectrophotometer was adjusted at 340 nm. The reagents were mixed and incubated at 25 °C for five minutes. The cuvettes were placed in the experimental and reference sites of spectrophotometer and recorded the change in absorbance for 3-4 minutes. There was 5.60% and 14.92% reduction in the activity of malate dehydrogenase in R. fruticosus and V. jatamansi, treated insects, respectively. The alpha amylase enzyme activity was 6.82% reduced and 63.63% increase in R. fruticosus and V. jatamansi, treated insects, respectively. Present study addresses that both plant powders are effective against granary weevil by altering enzyme activities so both the plant powders can be used as bio-pesticides against the stored grains pests.(AU)

As más condições de armazenamento proporcionam um ambiente favorável às pragas armazenadas para o crescimento. Os biopesticidas são as melhores alternativas aos pesticidas sintéticos. O presente estudo foi conduzido para comparar a toxicidade de Rubus fruticosus e Valeriana jatamansi contra gorgulhos, Sitophilus granarius e subsequentes alterações na atividade enzimática responsáveis por danos aos grãos. Na pesquisa atual, 5 g de frutos de R. fruticosus e pós de rizoma de V. jatamansi foram testados separadamente contra S. granarius, em 50 g de grãos integrais de trigo por sete dias, em comparação com o controle. A atividade enzimática da malato desidrogenase e alfa-amilase foi observada nos extratos celulares de S. granarius. Os insetos foram esmagados e homogeneizados em solução tampão fosfato e centrifugados a 10000 rpm por 5 minutos. Para a medição enzimática, o sobrenadante foi testado; o espectrofotômetro foi ajustado a 340 nm. Os reagentes foram misturados e incubados a 25 °C por cinco minutos. As cubetas foram colocadas nos locais experimentais e de referência do espectrofotômetro e registradas as alterações na absorbância por 3-4 minutos. Houve redução de 5,60% e 14,92% na atividade da malato desidrogenase em R. fruticosus e V. jatamansi, insetos tratados, respectivamente. A atividade da enzima alfa amilase foi reduzida em 6,82% e aumento de 63,63% em R. fruticosus e V. jatamansi, insetos tratados, respectivamente. O presente estudo aborda que ambos os pós de plantas são eficazes contra o gorgulho do celeiro, alterando as atividades enzimáticas, de modo que ambos os pós de plantas possam ser usados como biopesticidas contra pragas de grãos armazenados.(AU)

Sweet sorghum: broth clarification with enzymatic treatment increases the quality of the fermentation wort for ethanol production

Sweet sorghum is currently being evaluated throughout the world as a raw material for biofuel production because its stem juices are rich in sugars that can be directly fermented to ethanol. In this work, the fermentative efficiency of three sweet sorghum genotypes was evaluated, aiming at ethanol production, harvested in two seasons, clean and whole stems, and the treatment of the juice and broth with amylolytic enzymes in order to use the present starch to increase the production of ethanol. The experiment was carried out in the 2013/2014 harvest, in the municipality of Jaboticabal, São Paulo, Brasil, located at 21°14'05''S and 48°17'09''W. The experimental design was completely randomized, with sub-subdivided plots and four replications. The primary treatments were the sweet sorghum genotypes (CV147, CV198, and BRS508), the secondary treatments, the type of harvest (whole stems and clean stems); the tertiary the two sampling times (102 and 116 days after sowing - d.a.s) and the quaternary the application of enzymes. In the fermentation process, the yeast PE-2 was used, at the end, the wine was recovered and characterized. Fermentation efficiency and liters of ethanol per ton of sorghum were calculated. The clarification of the juice with enzymatic treatment increases the quality of the fermentation broth and makes it possible to obtain wines with lower levels of RRTs and Brix. Fermentation efficiency is not affected by the genotype; however, it is influenced by the time of harvest and the technological quality of the juice. The use of amylolytic enzymes makes it possible to obtain wines with lower levels of RRTS and Brix. The best period of industrialization was at 102 d.a.s., and the processing of whole stalks resulted in less ethanol production.

Salivary alpha amylase and cortisol levels as stress biomarkers in children with cerebral palsy and their association with a physical therapy program.

INTRODUCTION: Cerebral palsy (CP) is one of the main causes of physical disabilities in childhood. There is evidence that CP children display high levels of stress, which could interfere with learning processes and interpretation of relevant sensory information during motor skills acquisition and socialization. OBJECTIVE: This study aims to compare basal levels of stress biomarkers (cortisol and alpha-amylase) of healthy children (HC) and children with CP, and to investigate whether a physical therapy session using the neurodevelopmental technique (NDT) interferes with these levels. METHODS: A cross-sectional design was used. A total of 86 children (HC: n = 45 and CP: n = 41) with matching age, sex, socioeconomic status, and sampling time. Salivary cortisol and alpha-amylase levels were measured by means of electrochemiluminescence and spectrophotometry methods. A single saliva sample was collected in the HC group to determine basal levels. For CP group three samples were collected: a first sample was taken 20-30 min prior to the intervention, while two post-intervention samples were collected (5 and 20 min) to evaluate individual changes in salivary stress biomarkers. RESULTS: Higher basal cortisol concentration was found in CP children when compared to HC group. Moreover, CP children showed a significant reduction in cortisol levels 20 min after NDT intervention. No significant differences were observed in alpha-amylase values. CONCLUSION: Present results show that CP causes alteration in basal cortisol values at childhood and suggest that CP children respond to environmental regulatory factors such as NDT, in attempt to reduce stress.

Ameliorating effect of Malva Neglecta on hyperglycemia and hyperlipidemia in diabetic rats

The plant, Malva neglecta wallr., is widely consumed for medicinal and nutritional purposes. The current study was carried out to assess the hypoglycemic and antihyperlipidemic potential of aqueous methanolic extract of M. neglecta. Chemical evaluation of the extract was performed by high pressure liquid chromatography. Oral glucose tolerance test (OGTT) was done in diabetic rats pre-exposed to 250, 500 and 750 mg/kg plant extract via the oral route. For hypoglycemic and biochemical study, the same therapy was administered to alloxan induced diabetic rats for 14 days. The standard control group received Glibenclamide (5 mg/kg). Ferulic acid, p-coumaric acid and other phenolic acids were detected and estimated in the extract. Administration of the plant extract significantly reduced blood glucose level in diabetic rats subjected to OGTT. The plant extract lowered the fasting blood glucose and alpha amylase, and prevented the damage to pancreas. It also corrected dyslipidemia in diabetic animals following 14 days therapy. Hence, this experimental study establishes the fact that M. neglecta exhibited significant antidiabetic and antihyperlipidemic activities in alloxan induced diabetic rats.

Effects of A High Intensity Interval Session on Mucosal Immune Function and Salivary Hormones in Male and Female Endurance Athletes.

Although the effects of high intensity interval training (HIIT) on health and sports performance are well documented, the effects of this training type on mucosal immune function remain unclear. The aim of this study was to assess the impact of an acute HIIT session on salivary immune and endocrine marker levels (immunoglobulin A (sIgA), alpha amylase (sAA), cortisol (C), and testosterone (T)) in male and female endurance athletes. Twenty subjects (ten males and ten females) underwent ten bouts of treadmill running using a 4 min:2 min work:rest ratio at ~90% of peak oxygen uptake (VO2peak). Saliva samples were collected 5 min before and 20 min post-exercise. During work intervals, female participants had a higher HR than male participants (+4.0 ± 5%; p = 0.008). Rating of perceived exertion (RPE) increased throughout the duration of the HIIT session in both males and females (main time effect: p < 0.001), but was higher in males than females (+17 ± 4%; time x gender main effect: p < 0.001). Lactate concentrations were similar in both males and females. Exercise increased the concentration of salivary IgA (males: +24 ± 6%, p = 0.004; females: +27 ± 3%, p = 0.03), salivary alpha-amylase (males: +44 ± 22%, p = 0.036; females: +71 ± 26%, p = 0.026) and salivary cortisol (males: +41 ± 24%, p = 0.015; females: +55 ± 24%, p = 0.005). Testosterone levels and the Testosterone/Cortisol ratio remained stable in both males and females. These findings suggest that the physiological stress produced by a HIIT session does not affect immune function and does not disturb the anabolic/catabolic balance.

Comparison of protein profiles of the pellicle, gingival crevicular fluid, and saliva: possible origin of pellicle proteins.

BACKGROUND: The pellicle, the acellular organic material deposited on the surface of tooth enamel, has been thought to be derived from saliva. In this study, protein compositions of the pellicle, gingival crevicular fluid, and saliva collected from healthy adults were compared to elucidate the origin of pellicle proteins. RESULTS: The pellicle, gingival crevicular fluid, and saliva from the parotid gland or mixed gland were collected; subsequently, protein expression in samples from the respective individual was compared by SDS-PAGE and mass spectrometry. Following SDS-PAGE, proteins in the major bands were identified by mass spectrometry. The band pattern of pellicle proteins appeared different from those of gingival crevicular fluid, or saliva samples. Using mass spectrometry, 13 proteins in these samples were identified. The relative abundance of the proteins was quantitatively analyzed using mass spectrometry coupled with stable isotope labeling and by western blot. Cystatin S and α-amylase detected in pellicle were enriched in saliva samples, but not in gingival crevicular fluid, by western blot, and their abundance ratios were high in saliva and low in gingival crevicular fluid when analyzed by stable isotope labeling. Serotransferrin, however, was found only in the pellicle and gingival crevicular fluid by western blot and its abundance ratio was low in saliva. CONCLUSIONS: Our study revealed that the gingival crevicular fluid appears to contribute to pellicle formation in addition to saliva.

Horizontal Transfer and Gene Loss Shaped the Evolution of Alpha-Amylases in Bilaterians.

The subfamily GH13_1 of alpha-amylases is typical of Fungi, but it is also found in some unicellular eukaryotes (e.g., Amoebozoa, choanoflagellates) and non-bilaterian Metazoa. Since a previous study in 2007, GH13_1 amylases were considered ancestral to the Unikonts, including animals, except Bilateria, such that it was thought to have been lost in the ancestor of this clade. The only alpha-amylases known to be present in Bilateria so far belong to the GH13_15 and 24 subfamilies (commonly called bilaterian alpha-amylases) and were likely acquired by horizontal transfer from a proteobacterium. The taxonomic scope of Eukaryota genomes in databases has been greatly increased ever since 2007. We have surveyed GH13_1 sequences in recent data from ca. 1600 bilaterian species, 60 non-bilaterian animals and also in unicellular eukaryotes. As expected, we found a number of those sequences in non-bilaterians: Anthozoa (Cnidaria) and in sponges, confirming the previous observations, but none in jellyfishes and in Ctenophora. Our main and unexpected finding is that such fungal (also called Dictyo-type) amylases were also consistently retrieved in several bilaterian phyla: hemichordates (deuterostomes), brachiopods and related phyla, some molluscs and some annelids (protostomes). We discuss evolutionary hypotheses possibly explaining the scattered distribution of GH13_1 across bilaterians, namely, the retention of the ancestral gene in those phyla only and/or horizontal transfers from non-bilaterian donors.

Comparison of protein profiles of the pellicle, gingival crevicular fluid, and saliva: possible origin of pellicle proteins

BACKGROUND: The pellicle, the acellular organic material deposited on the surface of tooth enamel, has been thought to be derived from saliva. In this study, protein compositions of the pellicle, gingival crevicular fluid, and saliva collected from healthy adults were compared to elucidate the origin of pellicle proteins. RESULTS: The pellicle, gingival crevicular fluid, and saliva from the parotid gland or mixed gland were collected; subsequently, protein expression in samples from the respective individual was compared by SDS-PAGE and mass spectrometry. Following SDS-PAGE, proteins in the major bands were identified by mass spectrometry. The band pattern of pellicle proteins appeared different from those of gingival crevicular fluid, or saliva samples. Using mass spectrometry, 13 proteins in these samples were identified. The relative abundance of the proteins was quantitatively analyzed using mass spectrometry coupled with stable isotope labeling and by western blot. Cystatin S and α-amylase detected in pellicle were enriched in saliva samples, but not in gingival crevicular fluid, by western blot, and their abundance ratios were high in saliva and low in gingival crevicular fluid when analyzed by stable isotope labeling. Serotransferrin, however, was found only in the pellicle and gingival crevicular fluid by western blot and its abundance ratio was low in saliva. CONCLUSIONS: Our study revealed that the gingival crevicular fluid appears to contribute to pellicle formation in addition to saliva.