In Vitro Pancreatic Lipase, Alpha-Amylase and ?lpha-Glucosidase Inhibitory Activities of the Phytochemical Barbaloin

In the present study the phytochemical, barbaloin was studied for in-vitro pancreatic lipase, alpha (?)-amylase and alpha (?)-glucosidase inhibitory activities. The aim of this work is to evaluate the inhibitory activities of the phytochemical, barbaloin at different concentrations. Pancreatic lipase is an enzyme that hydrolyzes the lipids obtained from the diet which acts as an important target to treat obesity. The natural medicines that can inhibit pancreatic lipase enzyme and thus decrease absorption of dietary fat in the body gained much attention for the treatment and prevention of obesity. Diabetes mellitus is a metabolic disorder marked by an elevated level of glucose that circulates in the blood plasma. Alpha amylase and alpha glucosidase inhibitors are used to attain control over hyperglycemia in type 2 diabetes mellitus. The present study was designed to screen the novel pancreatic lipase, alpha amylase and alpha glucosidase inhibitors using a phytochemical, barbaloin in order to minimize the toxicity and side effects of the inhibitors which are used at present to treat the disorders like obesity and hyperglycemia. The phytochemical, barbaloin exhibited significant pancreatic lipase, ?-amylase and ?-glucosidase inhibitory activities with an IC50 value 5.52µg/ml, 8.22µg/ml and 5.81µg/ml respectively and well compared with standard orlistat for pancreatic lipase and acarbose for alpha (?)-amylase and alpha (?)-glucosidase inhibitory activities respectively.

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.

Effects of glucose as carbon catabolite repressor on alpha-amylase and glucoamylase production in Indonesian indigenous fungi

Aims@#The study aimed to investigate the effect of glucose on alpha-amylase and glucoamylase production in some Indonesian indigenous fungi.@*Methodology and results@#Fungi were screened for their ability to produce alpha-amylase and glucoamylase in the presence of glucose. The strains were grown in a medium containing starch and glucose as carbon sources with glucose concentrations varying from 0 to 5% for four days, and the alpha-amylase and glucoamylase were analyzed at the end of the growth period. Most strains showed repression on the amylases production when glucose was added to the medium. However, some strains showed no repression on amylases production when glucose was supplemented to the medium. The addition of glucose repressed glucoamylase production, but no repression on alpha-amylase was noted for strain KKB4, vice versa, there was repression on alpha-amylase production but no repression on glucoamylase production for strain FIG1. Strains FNCC 6151 and MLT1J1 showed no repression on both alpha-amylase and glucoamylase production when glucose was added to the medium up to 5%. The occurrence of repression in the production of alpha-amylase and glucoamylase was strain-specific.@*Conclusion, significance and impact of study@#Out of the nine indigenous fungi strains examined, strains FNCC 6151 and MLT1J1 showed no repression on both alpha-amylase and glucoamylase production when glucose was added to the medium up to 5%. Those two strains have the potential to be improved further to produce both alpha-amylase and glucoamylase.

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.

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.

Tylophora hirsuta L. leaf extract attenuates alloxan-induced diabetes in mice by suppressing oxidative stress and α-Amylase

Objective: To evaluate the antidiabetic potential of leaf extracts of Tylophora hirsuta (T. hirsuta). Methods: The methanolic and ethyl acetate extracts of T. hirsuta leaves were analyzed by high pressure liquid chromatography. In vitro antioxidant activity was determined by ferric ion reduction, 1, 1-diphenyl-2-picrylhydrazyl, and hydrogen peroxide scavenging methods. In vitro alpha amylase (α-Amylase) inhibitory activity of the plant extracts was assessed. In vivo antidiabetic potential was determined in alloxan-induced diabetic mice to assess glycated hemoglobin (HbA1c), oral glucose tolerance, serum amylase, lipid profile, fasting blood glucose, and body weight. Histopathological lesions of the pancreas, liver and kidney were observed. Oxidative stress biomarkers such as superoxide dismutase, catalase and peroxidase were also determined. Results: Quercetin, chlorogenic acid, p-coumaric acid, and m-coumaric acid were found in the plant extracts. The methanolic plant extract exhibited higher in vitro antioxidant activities than the ethyl acetate extract. Moreover, methanolic plant extract exhibited (83.90±1.56)% α-Amylase inhibitory activity at 3.2 mg/ mL concentration. Animal study showed that the methanolic extract of T. hirsuta improved the levels of fasting blood glucose, HbA1c, serum α-Amylase, lipid profile, liver function biomarkers, and kidney functions of diabetic mice. Moreover, the methanolic extract ameliorated diabetes-related oxidative stress by increasing superoxide dismutase and catalase activities and decreasing peroxidase and malondialdehyde levels. Histopathological examination showed that the plant extract had improved the integrity of pancreatic islets of Langerhans and reduced the pathological lesions in the liver and kidney of diabetic mice. Conclusions: The methanolic extract of T. hirsuta exhibits pronounced antidiabetic activity in mice through reduction of oxidative stress. The plant extract has several natural antioxidants such as phenolic acids. T. hirsuta extract could serve as a nutraceutical for managing diabetes mellitus.

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.

Effect Of Combination Of Aqueous Leaf Extracts Of Psidium Guajava Linn And Moringa Oleifera Lam On Diabetes Mellitus

Objective: To determine the anti-diabetic activity of combined aqueous extracts (1:1mixture) of dry leaves of Psidium guajava linn and Moringa oleifera lam as well as to compare the anti-diabetic activity of these plants by in vitro methods. Methods: In vitro alpha amylase inhibitory assay was performed on porcine alpha amylase and the absorbance was measured at 540 nm using a microplate reader and glucose diffusion inhibitory assay using dialysis membrane. Acarbose was used as the standard in the above mentioned methods. Results: The mixture (1:1) of aqueous plant extracts (at a concentration of 100µg/ml) of Psidium guajava linn and Moringa oleifera lam exhibited 72.08333% inhibition with IC50 value of 10.9µg/ml. The leaf extracts of Psidium guajava (at a concentration 100µg/ml) exhibited 71.23288% of a α amylase inhibitory activity with an IC50 values 19.883µg/ml whereas the leaf extracts of Moringa oleifera (at a concentration of 100µg/ml) exhibited 70.58824% of α amylase inhibitory activity with an IC50 value of 27.974 µg/ml. The Acarbose (standard drug) at a concentration of 100µg/ml showed 72.09302% inhibitory effect on the α amylase activity with an IC50 value 8.9µg/ml. In glucose diffusion inhibition assay the mixture of plant extracts exhibited 76.57% inhibition at 150 min which produces more effects than the two plants. The aqueous extract of Psidium guajava leaves exhibited maximum glucose diffusion inhibition (75.32%) at 150 min as well as Moringa oleifera leaf extract showed the maximum inhibition of 73.70% at the same time interval. For acarbose the percentage was 82.74 at 150 min. The interpretation of the results was done by one-way anova method. Conclusion: The combined extract of the leaves of the 2 plants was found to be more effective than individual plant extracts against diabetes. On comparison of two plants Psidium guajava was found to be more active against diabetes than Moringa oleifera. Also the potentiation effect shown by the combination of extract may be due to synergistic effect of the phytochemical constituents. As the 1:1 mixture of the aqueous extract is found to be more active, the combination of the two plants can be used to formulate drugs for treating diabetes.

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.

In vitro anti-amylase activity of some Indian dietary spices

Pancreatic alpha amylase (PAA) inhibitory activity of cinnamon, cumin was evaluated in vitro to search newanti-diabetic agents as alternatives to synthetic medicines. Bark of cinnamon, seeds of cumin, fenugreek,nutmeg, fennel, and buds of clove were extracted with hot water, methanol, chloroform, benzene, and ethylacetate and 30 extracts were tested for the presence of PAA inhibitory activity using qualitative and quantitativemethods and their modes of inhibition were determined. Presence of alpha amylase inhibitors was identified in18 extracts in quantitative assay. Benzene extracts of cinnamon, clove, fenugreek, and nutmeg and chloroformextract of cumin showed highest anti-amylase potential. The IC50 values of these potential extracts rangedbetween 2.09 ± 0.12 mg/ml and 2.89 ± 0.079 mg/ml with lowest IC50 value noted for benzene extract ofcinnamon and highest IC50 value was noted for benzene extract of fenugreek. Based on the values of Vmaxand Km compared to control, it was found that all these extracts display non-competitive mode of inhibition onamylase activity. In conclusion, active constituents of these five extracts possess anti-amylase properties andcan be used in management of diabetes mediated complications.

Salivary alpha-amylase as a stress biomarker in diseased dogs

Salivary alpha-amylase (sAA) is a stress biomarker in human diseases, but there are no reports of sAA measurements in diseased dogs. This study measured the sAA and serum alpha-amylase (AA) levels in 16 healthy dogs and 31 diseased dogs using a kinetic enzyme assay to assess the stress status. The sAA and serum AA levels were significantly higher in the diseased dogs than in healthy dogs (p < 0.05), but there was no correlation between the 2 groups (r = 0.251, p = 0.089). This suggests that sAA can be useful as a stress biomarker in diseased dogs.

Relationship between Personality and Biological Reactivity to Stress: A Review

OBJECTIVE: Personality traits can be the basis for individual differences in the biological response of stress. To date, many psychobiological studies have been conducted to clarify the relationship between personality and biological reactivity to stress. This review summarizes the most important findings in this area of research. RESULTS: Key findings related to the relationship between personality factors and stress-sensitive biological systems in four research models have been summarized; model of psychosocial characteristics, model based on Rumination and Emotional Inhibition, Eysenck’s biopsychological model, and Five-Factor Approach of Personality. CONCLUSION: According to the results of this review, it can be concluded that personality typology of individuals influenced their biological reactivity to stressful events. Understanding the biological basis of personality can help to better understand vulnerability to stress. Future research can be continuing based on framework of the four models.

Determinação da energia metabolizável do farelo residual do milho com e sem enzima em dietas para frangos de corte / Determination of the metabolizable energy of residual corn bran with and without enzyme in diets for broilers

O objetivo deste trabalho foi avaliar o valor nutricional e determinar a energia metabolizável do farelo residual de milho (FRM) sem e com o uso da enzima alfa- amilase. Foi realizado um experimento de metabolismo com 180 pintos machos Cobb com 14 dias, distribuídos em um delineamento inteiramente ao acaso, com seis tratamentos, cinco repetições e seis aves por parcela. As dietas experimentais foram: T1: ração referência (RR), T2: 60% T1 + 40% de FRM, T3: RR + enzima, T4: 60% T1 + 40% de FRM com adição de enzima, T5: RR com substituição de 100% do milho pelo FRM e T6: RR com substituição de 100% do milho pelo FRM com adição de enzima. A composição química do FRM foi: 88,33% de matéria seca (MS), 10,23% de proteína bruta (PB), 15,44% de extrato etéreo (EE), 4,33% de cinzas (CZ) e 4555kcal/kg de energia bruta (EB). Os valores dos coeficientes de metabolizabilidade aparente para o FRM sem e com adição de enzima foram, respectivamente, de 73,37% e 76,33% para MS (p=0,0136), 70,44% e 70,39% para PB (p=0,9595) e de 74,79% e 76,77% para EB (p=0,0128). Os valores da energia metabolizável aparente (EMA) e da EMA corrigida para retenção de nitrogênio (EMAn) para o FRM (na base natural) foram de 3322±19 e 3241±18kcal/kg e de 3334±16 e 3261±17kcal/kg, respectivamente, sem e com adição de enzima. A adição da enzima não teve efeito estatístico significativo sobre os valores de EMA e EMAn, entretanto houve efeito positivo no coeficiente de metabolização da energia.(AU)

The aim of this study was to evaluate the nutritional value and the metabolizable energy of residual corn bran (FRM) with and without the use of alpha amylase enzyme. One metabolism assay was done using 180 Ross male broilers at 14 days of age distributed in an entirely randomized design with six treatments, five replications and six broilers per plot. The assay treatments established were: T1: reference diet (RR), T2: 60% T1 + 40% of FRM, T3: T1 (RR) + enzyme addition, T4: 60% T1 + 40% of FRM with enzyme addition, T5: T1 (RR) with 100% replacement of corn by FRM and T6: T1 (RR) with 100% replacement of corn by FRM with enzyme addition. FRM chemical composition was: 88.33% of dry matter (MS), 10.23% of crude protein (PB), 15.44% of crude fat (EE), 4.33% of ashes (CZ) and 4555Kcal/kg of Gross Energy (EB). The FRM apparent metabolizability coefficients with and without the enzyme addition were, respectively, 73.37% and 76.33% for MS (p=0.0136), 70.44% and 70.39% for PB (p=0.9595) and 74.79% and 76.77% for EB (p=0.0128). The apparent metabolizable energy (EMA) and EMA corrected for nitrogen retention values (as is basis) were 3322±19 and 3241±18Kcal/kg and 3334±16 and 3261±17Kcal/kg, respectively, for with and without the enzyme addition to FRM. The enzyme addition had no statistical significance on the EMA and EMAn values, however, a significant effect on energy metabolizability coefficient was observed.(AU)

The changes and the significance of salivary levels of immunoglobulin A, alpha-amylases and acetylcholinesterase in patients with Parkinson disease / 中华老年医学杂志

Objective To detect the salivary levels of IgA,alpha-amylase and acetylcholinesterase (AChE) and saliva flow rates of IgA in normal controls versus Parkinson disease (PD) patients,and to analyze the correlation of these salivary biochemical indexes with disease duration,H&Y rating scale,SCOPA-AUT score,sialorrhea score,dry mouth score and the use of levodopa.Methods Thirty-eight PD patients including twenty-two patients treated with levodopa,and twenty-one normal controls were selected in our study.About 3 ml saliva was collected from each subject.Salivary IgA was detected by immunoturbidimetry.Alpha-amylase and AChE were tested by enzymatic colorimetry.The protein concentration was analyzed by BCA kit to normalize ACHE.The salivary flow rate and IgA flow rate were also calculated.Results Salivary flow rate (ml/min) was (0.39±0.22) in normal controls,and (0.35±0.28) in PD groups;the level of IgA (mg/L) was (148.3±86.1) in normal controls,and (183.7±126.3) in PD groups;IgA flow rate (μg/min) was (47.8±25.9) in normal controls,and (46.9±27.1) in PD groups;the level of alpha-amylase (U/L) was (1.63 ± 1.21) × 105 in normal control,and (1.35 ± 9.87) × 105 U/L in PD group;and the level of AChE (U/mg) was (0.12±0.08) in normal controls,and (0.09±0.05) in PD group.No statistically significant differences in above indexes were found between PD group and controls.There were negative correlations between salivary flow rate and dry mouth score (r =-0.445,P =0.005) and between alpha-amylase concentration and sialorrhea score (r=-0.327,P=0.045).The positive associations were found between IgA concentration and the SCOPA-AUT score (r =0.438,P =0.006),and between IgA flow rate and sialorrhea score (r =0.411,P =0.01).Conclusions Salivary flow rate and level of alpha-amylases tend to be decreased in PD patients,while IgA concentration has an upward tendency.Salivary alpha-amylases concentration and IgA flow rate are correlated with the symptom of sialorrhea,while IgA concentration correlates with the degree of autonomic symptoms.These measurements can be used as indexes for evaluating the lesions of the autonomic nervous system.

Influence of Pre-treatment on Secondary Metabolites and Hypo-Glycemic Activity of Custard Apple (Annona squamosa) Peel

Plant secondary metabolites, present in the outer layers of fruits and vegetables in higher amounts, are structurally diverse and comprise different classes of phyto-constituents that have a number of health-promoting properties. Blanching is an important pre-treatment that is used to inactivate the enzymatic oxidation. Methods: This study was designed to investigate the effects of steam blanching (3, 5 and 7 min) and drying (50ºC) of custard apple (Annona squamosa) peel on the quantification of secondary metabolites and hypo-glycemic activity. Unblanched sample dried at 50ºC served as control. Quantitative tests for alkaloids, tannins, saponins and flavonoids were carried out for all samples and in vitro alpha amylase inhibition test was done to confirm the hypo-glycemic properties. All analyses were done in triplicates. Results: Quantitative results for alkaloids, tannins, saponins showed a significant reduction (p0.05). The exception was observed in the flavonoid content which showed a significant increase for 3 min and 5 min blanched samples, while the 7 min blanched sample showed a reduction in flavonoid content over the unblanched and fresh samples. Alpha amylase inhibition test similarly showed a decreasing trend for blanched samples ranging from IC50 value of 3.31 to 5.53 μg/mL compared to the unblanched with IC50 value of 4.92 μg/mL and fresh sample with IC50 value of 6.37μg/mL. Conclusion: From the study, it is inferred that steam blanching and drying have a significant impact on the quantification of secondary metabolites and subsequently on its hypo-glycemic activity. A steam blanching time of 5 min is the optimum for processing of custard apple peel.

Study on effects of citric acid induced acid load on salivary secretion / 国际检验医学杂志

Objective To investigate the effects of citric acid on the related indexes of salivary secretion under acid loading in or‐der to optimize the citric acid load method .Methods The saliva samples were collected from 10 young healthy volunteers at 1-90 s before ,at 91-120 s during and at 121-210 s after citric acid loading .The indexes were detected in saliva with mixed loading and after loading .The salivary alpha‐amylase(sAA ) activity ,pH value ,saliva flow rate ,total protein concentration in various groups were detected .The ratio values before and after the acid loading were compared among the groups .Results (1)The sAA activity , saliva pH value and total protein concentrations after acid loading were significantly increased compared before loading (P<0 .05) , moreover the ratio of after loading and before loading was greater than 1(P<0 .05);(2) however in the citric acid mixing ,the sAA activity ,saliva pH value and total protein concentration were decreased compared with before acid loading ,its ratio was less than 1 (P<0 .05) .Conclusion Citric acid affects the secretion result of acid loading saliva secretion ,it is suggested that the saliva under acid loading is separated treated and analyzed .

Evaluation of salivary and serum alpha amylase level in dentalcaries of adolescence / Avaliação do nível de alfa-amilase salivar e sérica na cárie dentária da adolescência

Previous studies suggested a significant relationship between alpha- amylase and caries formation. This study was implemented in order to investigate the interrelation between level of salivary and serum alpha- amylase, and dental caries. Methods: In this cross-sectional study, un-stimulated whole saliva and serum samples were collected from 118 high school students who were divided into four groups: caries free females (N = 28), caries active females (N = 35), caries free males (N = 28) and caries active males (N =27). Mean levels of salivary and serum alpha-amylase was evaluated by spectrophotometric methods to assayenzyme kinetics. Data were analyzed using student’st-tests and chi-square tests. Results: Salivary alpha amylase was significantly higher in caries active groups as compared to caries free groups (P = 0.002). The mean salivary alpha -amylase was significantly higherin caries active males (P = 0.002). Also, serum alpha-amylase was significantly higher in caries active groups as compared to caries free groups (P = 0.001). In bothsexes the mean serum alpha- amylase levels were statistically significant higher in caries active compared to caries free groups (P = 0.02 and 0.01 respectively). Conclusion: A significant association was found between salivary and serum alpha- amylase and dental caries in adolescents. More research to demonstrate there al relation between alpha amylase and dental caries is recommended...

Objetivo: Alguns estudos anteriores sugeriram uma relação significativa entre a alfa-amilase e a formação de cárie. Este estudo foi realizado com o objetivo de investigar a interrelação entre o nível de alfa-amilase na saliva e no soro e a cárie. Material e Métodos: Neste estudo transversal, amostras de saliva não-estimulada e soro foram coletadas de 118 estudantes do ensino médio que foram divididos em quatro grupos: mulheres livres de cárie (n = 28), mulheres com cárie ativa (n = 35), homens livres de cárie (n = 28) e homens com cárie ativa (n = 27). O nível de alfa-amilase da saliva e soro foi avaliado por espectrofotometria para ensaios de cinética enzimática. Os dados foram analisados pelos testes “t”e Qui-quadrado. Resultados: A alfa-amilase salivar foi significativamente maior nos grupos com cárie ativa em comparação com os grupos livres de cárie (p = 0,002). A alfa amilase salivar foi significativamente maior nos homens com cáries ativa (p = 0,002). Além disso, a alfa amilase sérica foi significativamente maior nos grupos de cárie ativa, em comparação com os grupos livres de cárie(p = 0,001). Em ambos os sexos a média da alfa-amilase sérica foi estatisticamente superior nos grupos de cárie ativa, em comparação com livres de cárie (p = 0,02 e 0,01, respectivamente). Conclusão: Foi encontrada uma associação significativa entre a alfa-amilase salivar e sérica em indivíduos com a cárie dentária na adolescência. São recomendadas mais investigações para demonstrar real relação entre os níveis de alfa-amilase e cárie dentária...

Evaluación de estrés percibido y niveles de alfa-amilasa salival en mujeres embarazadas (del Municipio Libertador del Estado Mérida) / Evaluation of perceived stress and levels of salivary alpha-amylase in pregnant women (the Libertador Municipality of Mérida State)

OBJETIVO: Investigar la percepción del estrés en mujeres embarazadas y su interrelación con los niveles de la enzima alfa amilasa salival (AAS) que es un marcador biológico de estrés. MÉTODOS: Incluimos 38 embarazadas que acudían a la consulta prenatal en Centros de Salud del Municipio Libertador del Estado Mérida, Venezuela. Se distribuyeron en 3 grupos de acuerdo con la edad gestacional, 8 en el primer trimestre, 10 en el segundo, 10 en el tercer y como grupo control se tomaron 10 mujeres no embarazadas con características demográficas similares. El estrés percibido se midió a través del instrumento EEP-13 y el nivel de enzima alfa amilasa salival mediante ensayo enzimático con espectrofotometría. Con los puntajes del EEP-13, se establecieron tres niveles de estrés percibido. RESULTADOS: El 23,7 % de la muestra tenía niveles bajos de estrés percibido, 50 % poseía un nivel intermedio y la cuarta parte (26,3 %) mostró un nivel alto. No se encontró relación significativa entre el nivel de estrés percibido y la etapa del embarazo (P = 0,637). La concentración de enzima alfa amilasa salival fue significativamente mayor en el grupo de embarazadas en el primer trimestre en comparación con el control (P<0,001), sin embargo, no encontramos correlación entre la puntuación del EEP-13 (R²=0,014; P=0,81) y los niveles de enzima alfa amilasa salival. CONCLUSIÓN: El 76,3 % de este grupo de embarazadas mostró niveles significativos de estrés según el EEP-13 y solo el grupo de embarazadas en el primer trimestre mostró niveles de enzima alfa amilasa salival superiores al grupo control.

OBJECTIVE: To assess the levels of salivary alpha amylase enzyme (SAA) that is a biomarker of stress and the correlation of those levels with the perception of stress in pregnant women. METHODS: Thirty eight pregnant women who attended prenatal care clinics in the Libertador Municipality of Merida, Venezuela, were distributed in 3 groups according with the gestational age: 8 on the first trimester, 10 on the second and 10 on the third. The control group consisted of 10 non-pregnant women with similar demographic characteristics. The perceived stress was measured with the EEP-13 instrument and the level of SAA was determined by means of an spectrophotometric enzymatic assay. Three levels of perceived stress were established according to the score obtained by the EEP-13 test. RESULTS: Twenty three percent (23.7 %) of the pregnant women had low levels of perceived stress, 50 % had an intermediate level and 26.3 % showed a high level. No significant relationship between the level of perceived stress and the age of pregnancy was found (P=0.637). The SAA concentration was significantly higher in the group of pregnant women in the first trimester compared to the non-pregnant control group (P <0.001). No correlation between the EEP-13 test values of perceived stress and the enzyme concentration was found (R2=0.014; P=0.81). CONCLUSION: In this group of pregnant women, 76.3 % suffer from a significant amount of stress and only the pregnant women in the first trimester showed increased levels of SAA.

Inhibitory effects of an extract of fruits of Physalis peruviana on some intestinal carbohydrases / Evaluación in vitro del efecto de un extracto de frutos de Physalis peruviana sobre algunas carbohidrasas intestinales

Physalis peruviana is an Andean specie whose fruits are eaten as food and also has been reported in Colombian folk medicine for diabetes mellitus treatment. In addition, previous pharmacological studies on diabetic Wistar rats, an extract of P. peruviana fruits has been showed antidiabetic activity. In order to deepen in P. peruviana action mode as antidiabetic, in this investigation were determinate the Inhibitory Concentrations 50 (IC50) of P. peruviana fruits crude extract on alpha glucosidase from S. cerevisiae and from intestinal rat powder, on maltase and alpha amylase enzymes. The kinetic behavior of the extract on each enzyme was also investigated, and the enzyme constant (Km) and maximum rate (Vmax) were determined. Extract of fruits of P. peruviana showed different IC50 for alpha glucosidase from S. cerevisiae and intestinal rat powder, suggesting greater affinity for the enzyme of mammalian source (4114.7 and 3552.7 µg/mL, respectively). For maltase, the IC50 was close to that obtained for alpha glucosidase (4191.0 µg/mL) while for alpha amylase, the extract exhibited the highest inhibition (IC50: 619.9 g/mL). Regarding kinetic behavior, the extract showed competitive inhibition on alpha-glucosidase and maltase, and on the non-competitive type of alpha amylase. These suggest that inhibition of intestinal carbohydrases is one of the modes of action for the antidiabetic activity of fruits of P. peruviana.

Physalis peruviana es una especie andina, cuyos frutos además de ser usados como alimento, son empleados en la medicina tradicional para el tratamiento de la diabetes mellitus. Además, estudios farmacológicos previos en ratas Wistar han demostrado actividad antidiabética de extractos de frutos de P. peruviana. Con el fin de profundizar en el modo de acción de la actividad antidiabética de los frutos de P. peruviana, en la presente investigación se determinó la concentración inhibitoria 50 (CI50) del extracto crudo de frutos de P. peruviana, sobre las enzimas alfa glucosidasa obtenida de S. cerevisiae y de polvo intestinal de rata, maltasa y alfa amilasa. El comportamiento cinético del extracto sobre cada una de las enzimas también fue investigado y la constante enzimática (Km) y la velocidad máxima (Vmax) fueron determinadas. El extracto de frutos de P. peruviana, mostró diferentes valores de CI50 para alfa glucosidasa obtenida de S. cerevisiae y para la obtenida de polvo intestinal de rata, sugiriendo una mayor afinidad por la enzima de origen mamífero (4114,7 and 3552,7 µg/mL, respectivamente). Para maltasa la CI50 fue cercana a la obtenida para alfa glucosidasa (4191,0 µg/mL), mientras para alfa amilasa, el extracto presentó la mayor inhibición (CI50: 619,9 g/mL). Respecto al comportamiento cinético, el extracto mostró inhibición de tipo competitiva sobre alfa glucosidasa y maltasa y no competitiva sobre alfa amilasa. Los resultados sugieren que la inhibición de carbohidrasas intestinales es uno de los modos de acción de los frutos de P. peruviana como agente antidiabético.

Carbohydrate hydrolyzing enzyme inhibitor property, antioxidant and phytochemical analysis of Cassia auriculata, Delonix regia and Vinca rosea Linn: an in vitro study.

The study comprises the evaluation of antidiabetic and antioxidant activities of Cassia auriculata, Delonix rega and Vinca rosea belonging to Fabaceae family. Initially, In vitro α-Glucosidase inhibitor activity was performed as a preliminary screening for petroleum ether, chloroform, ethyl acetate, methanol and aqueous extracts of three collected plants. In comparison with all the extracts, methanol extracts shown promising activity with IC50 values of 58.52μg/ml, 83.46μg/ml and 77.41μg/ml for C. auriculata, D. regia and V. rosea respectively. Hence, these three extracts were further selected for DPPH radical scavenging activity. C. auriculata resulted in 96% DPPH radical scavenging activity fallowed by D. regia (78%) and V. rosea (60%). Hence, methanol extract of C. auriculata was selected to evaluate α-amylase inhibitor potential, reducing power capacity and also quantified for total phenolic and total flavonoid content within them. Selected extract showed efficient α-amylase inhibitor activity, reducing power capacity and good amount of phenolics and flavonoids. HPTLC analysis identified five polyphenols and FT-IR analysis detected –OH functional groups in the extract. GC-MS analysis detected hydroquinone, myoionositol and bulletin. It can be concluded that these compounds within the methanol extract of C. auriculata might be accountable for antidiabetic and antioxidant nature of the plant. In future, bioactive compound could be isolated and used as a carbohydrate hydrolyzing enzyme inhibitor to treat type 2 diabetic individuals.