A colorimetric assay and MCR-ALS analysis of the peroxidase-like activity of poly (N-phenylglycine) functionalized with polyethylene glycol (PNPG-PEG) nanozyme for the determination of dopamine.

This report describes, for the first time, the coupling of UV-visible spectroscopy with multivariate curve resolution-alternative least-squares (MCR-ALS) algorithm to study peroxidase-like catalytic reaction of polyethylene glycol-functionalized poly (N-phenyl glycine) (PNPG-PEG) as an efficient and intrinsic peroxidase mimic activity (PMA) class of conducting organic polymer for selective detection of dopamine (DA) in the PNPG-PEG + TMB + H2O2 reaction system. PNPG-PEG was produced by means of a chemical route using ammonium persulphate (APS) as an oxidant agent of N-phenyl glycine monomer. The chemical composition, morphology, and thermal behavior of PNPG-PEG were examined by various instrumental techniques. PNPG-PEG exhibited significant peroxidase-mimic activity to catalyze the oxidation 3,3',5,5'- tetramethylbenzidine (TMB) substrate to oxidized TMB (oxTMB). The qualitative and quantitative determination of the oxidized TMB can easily be detected by the naked-eye and the recorded UV-vis absorbance spectra at 652 nm, respectively. Owing to the superior peroxidase-mimic activity of PNPG-PEG, the colorimetric detection of dopamine was successfully achieved at pH 4.0. Under optimal conditions, acceptable linear dependency was recorded in the concentration range of 5.1-125.0 µM, with a limit of detection (LOD) and limit of quantification (LOQ) equal to 4.6 µM and 13.8 µM (S/N = 3), respectively. Furthermore, this colorimetric assay was successfully used for quantitative analysis of dopamine in fetal bovine serum (FBS) and horse serum (HS) samples with recoveries in the range of 97-105% and 100-122%, respectively. After resolving the bilinear data matrix using MCR-ALS, three chemical components were found for different concentrations and pure spectral profiles. Based on the resolved profiles, the presence of free, slightly penetrated, and majorly penetrated TMB molecules entering the polymeric structure can be easily detected using MCR-ALS as an available statical method without any complex separation instruments. This peroxidase mimetic nanozyme as a visual, simple, low-cost, sensitive, and reproducible colorimetric platform can provide great potential applications in the monitoring and diagnosis of dopamine-related diseases.

Effect of various drying methods on the physicochemical characterizations, antioxidant activities and hypoglycemic activities of lychee (Litchi chinensis Sonn.) pulp polysaccharides.

Suitable drying method is critical for the preservation of physicochemical and pharmacological quality of lychee pulp polysaccharides (LPPs). In current work, the effects of five drying methods, i.e. air drying (A), infrared drying (I), heat pump drying (H), vacuum freeze drying (F) and freeze vacuum drying combined with heat pump drying (FH) on the physicochemical characterizations, antioxidant activities and hypoglycemic activities of LPPs were explored. Results showed all five drying methods led to thermal aggregation of LPPs and the stronger the thermal effect induced by drying, the more serious the aggregations were. Additionally, the thermal aggregation significantly affected the composition, structure and biological activity of LPPs. Less thermal aggregation was observed in LPPF and LPPFH, which exhibited stronger oxygen, DPPH and ABTS radical scavenging activities, higher ferric-reducing power and better α-glycosidase and α-amylase inhibition activities, resulting from their higher contents of neutral sugar, protein and uronic acid and lower molecular weight than LPPA and LPPI. Besides, FH consumed about half drying time and one fifth energy of F. Therefore, from industrial perspective, FH is a promising alternative to F for producing LPPs by comprehensively considering physicochemical characterizations, bioactivity as well as energy consumption.

Exploring the catalytic mechanism of a novel ß-glucosidase BGL0224 from Oenococcus oeni SD-2a: Kinetics, spectroscopic and molecular simulation.

The ß-glucosidase derived from microorganisms has attracted worldwide interest for their industrial applications, but studies on ß-glucosidases from Oenococcus oeni are rare. In this paper, catalytic mechanism of a novel ß-glucosidase BGL0224 of Oenococcus oeni SD-2a was explored for the first time by kinetic parameters determination, fluorescence spectroscopy and quenching mechanism analysis, molecular dynamics simulation. The results indicated that BGL0224 had universal catalytic effect on different types of glycoside substrates, but the catalytic efficiencies were different. Fluorescence quenching analysis results suggested that the quenching processes between BGL0224 and seven kinds of substrates were predominated by the static quenching mechanism. A reasonable three-dimensional model of BGL0224 was obtained using the crystal structure of E.coli BglA as a template. The analysis results of molecular simulation (RMSD, Rg, RMSF and hydrogen bonding) showed that the composite system 'BGL0224-pNPG' was very stable after 40 ns. The catalytic process of BGL0224 acting on 'p-Nitrophenyl ß-d-glucopyranoside' conformed to the double displacement mechanism. Two glutamic acid residues 'Glu178 and Glu377' played a vital role in the whole catalytic process. Overall, this study gave specific insights on the catalytic mechanism of BGL0224, which was of great significance for developing its potential applications in food industry.

Comparison of edible brown algae extracts for the inhibition of intestinal carbohydrate digestive enzymes involved in glucose release from the diet.

Type II diabetes is considered the most common metabolic disorder in the developed world and currently affects about one in ten globally. A therapeutic target for the management of type II diabetes is the inhibition of α- glucosidase, an essential enzyme located at the brush border of the small intestinal epithelium. The inhibition of α-glucosidase results in reduced digestion of carbohydrates and a decrease in postprandial blood glucose. Although pharmaceutical synthetic inhibitors are available, these are usually associated with significant gastrointestinal side effects. In the present study, the impact of inhibitors derived from edible brown algae is being investigated and compared for their effect on glycaemic control. Carbohydrate- and polyphenolic-enriched extracts derived from Ascophyllum nodosum, Fucus vesiculosus and Undaria pinnatifida were characterised and screened for their inhibitory effects on maltase and sucrase enzymes. Furthermore, enzyme kinetics and the mechanism of inhibition of maltase and sucrase were determined using linear and nonlinear regression methods. All tested extracts showed a dose-dependent inhibitory effect of α-glucosidase with IC50 values ranging from 0⋅26 to 0⋅47 mg/ml for maltase; however, the only extract that was able to inhibit sucrase activity was A. nodosum, with an IC50 value of 0⋅83 mg/ml. The present study demonstrates the mechanisms in which different brown seaweed extracts with varying composition and molecular weight distribution differentially inhibit α-glucosidase activities. The data highlight that all brown seaweed extracts are not equal in the inhibition of carbohydrate digestive enzymes involved in postprandial glycaemia.

Antidiabetic effects of Maclura cochinchinensis (Lour.) corner heartwood extract.

BACKGROUND AND AIM: Maclura cochinchinensis (MC) (Lour.) heartwood extract have been used traditionally to treat diabetes in Thailand, but their mechanism of action has not been elucidated. EXPERIMENTAL PROCEDURE: This study investigated the effects of an aqueous heartwood extract of MC on α-glucosidase and α-amylase activities. Moreover, its antidiabetic effect was evaluated using an oral glucose tolerance test (OGTT) and a 28-day administration to streptozotocin (STZ)-nicotinamide (NA)-induced type 2 diabetic mice. RESULTS: In both OGTT and the daily oral administration for 28 days in STZ-NA-induced type 2 diabetic mice models, the extract (1,000 mg/kg) significantly decreased fasting blood glucose. This hypoglycemic effect was explained by increased insulin levels and α-glucosidase inhibition (IC50:1.53 ±â€¯0.03 µg/mL). CONCLUSION: This first study on the hypoglycemic activity of MC heartwood extract indicates that it could be a potential natural remedy for type 2 diabetes.

Enzyme kinetics and inhibition parameters of human leukocyte glucosylceramidase.

Glucosylceramidase (GCase) is a lysosomal enzyme that catalyzes the cleavage of ß-glucosidic linkage of glucocerebroside (GC) into glucose and ceramide; thereby, plays an essential function in the degradation of complex lipids and the turnover of cellular membranes. The growing list of 460 mutations in the gene coding for it-glucosylceramidase beta acid 1 (GBA1)-is reported to abolish its catalytic activity and decrease its enzyme stability, associating it with severe health conditions such as Gaucher disease (GD), Parkinson Disease (PD) and Dementia with Lewy bodies (DLB). Although the three-dimensional structure of wild type glucosylceramidase is elucidated, little is known about its features in human cells. Moreover, alternative sources of GCase that prove to be effective in the treatment of diseases with enzyme treatment therapies, impose the need for a simple and cost-effective procedure to study the enzyme behavior. This work, for the first time, shows a well-established, yet simple, cost- and time-efficient protocol for the study of GCase enzyme in human leukocytes by the artificial substrate p-Nitrophenyl-ß-D-glucopyranoside (PNPG). Characterization of the enzyme in human leukocytes for activation parameters (optimal pH, Km, and Vmax) and enzyme inhibition was done. The results indicate that the optimum pH of GCase enzyme with PNPG is 5.0. The Km and Vmax values are 12.6mM and 333 U/mg, respectively. Gluconolactone competitively inhibits GCase, with a Ki value of 0.023 mM and IC50 of 0.047 mM. Glucose inhibition is uncompetitive with a Ki of 1.94 mM and IC50 of 55.3 mM. This is the first report for the inhibitory effect of glucose, δ-gluconolactone on human leukocyte GCase activity.

Designing chimeric enzymes inspired by fungal cellulosomes.

Cellulosomes are synthesized by anaerobic bacteria and fungi to degrade lignocellulose via synergistic action of multiple enzymes fused to a protein scaffold. Through templating key protein domains (cohesin and dockerin), designer cellulosomes have been engineered from bacterial motifs to alter the activity, stability, and degradation efficiency of enzyme complexes. Recently a parts list for fungal cellulosomes from the anaerobic fungi (Neocallimastigomycota) was determined, which revealed sequence divergent fungal cohesin, dockerin, and scaffoldin domains that could be used to expand the available toolbox to synthesize designer cellulosomes. In this work, multi-domain carbohydrate active enzymes (CAZymes) from 3 cellulosome-producing fungi were analyzed to inform the design of chimeric proteins for synthetic cellulosomes inspired by anaerobic fungi. In particular, Piromyces finnis was used as a structural template for chimeric carbohydrate active enzymes. Recombinant enzymes with retained properties were engineered by combining thermophilic glycosyl hydrolase domains from Thermotoga maritima with dockerin domains from Piromyces finnis. By preserving the protein domain order from P. finnis, chimeric enzymes retained catalytic activity at temperatures over 80 °C and were able to associate with cellulosomes purified from anaerobic fungi. Fungal cellulosomes harbor a wide diversity of glycoside hydrolases, each representing templates for the design of chimeric enzymes. By conserving dockerin domain position within the primary structure of each protein, the activity of both the catalytic domain and dockerin domain was retained in enzyme chimeras. Taken further, the domain positioning inferred from native fungal cellulosome proteins can be used to engineer multi-domain proteins with non-native favorable properties, such as thermostability.

Dual substrate/solvent- roles of water and mixed reaction-diffusion control of ß-Galactosidase catalyzed reactions in PEG-induced macromolecular crowding conditions.

Here we studied the effect of molecular crowding on the hydrolysis of ortho- and para-nitrophenyl-ß-D-galactopyranosides (ONPG, PNPG) catalysed by Escherichia coli ß-Galactosidase in the presence of 0-35%w/v 6kD polyethyleneglycol (PEG6000). The Eadie-Hofstee data analysis exhibited single straight lines for PNPG at all [PEG6000] as well as for ONPG in the absence of PEG6000 so a Michaelian model was applied to calculate the kinetic parameters KM and kcat (catalytic rate constant) values. However, for ONPG hydrolysis in the presence of PEG6000, the two slopes visualized in Eadie-Hofstee plots leaded to apply a biphasic kinetic model to fit initial rate vs. [ONPG] plots hence calculating two apparent KM and two kcat values. Since the rate limiting-step of the enzymatic hydrolysis mechanism of ONPG, but not of PNPG, is the water-dependent one, the existence of several molecular water populations differing in their energy and/or their availability as reactants may explain the biphasic kinetics in the presence of PEG6000. With PNPG, KM as well as kcat varied with [PEG6000] like a parabola opening upward with a minimum at 15 %w/v [PEG6000]. In the case of ONPG, one of the components became constant while the other component exhibited a slight increasing tendency in kcat plus high and [PEG6000]-dependent increasing KM values. Sedimentation velocity analysis demonstrated that PEG6000 impaired the diffusion of ß-Gal but not that of substrates. In conjunction, kinetic data reflected complex combinations of PEG6000-induced effects on enzyme structure, water structure, thermodynamic activities of all the chemical species participating in the reaction and protein diffusion.

Bioaccessibility and inhibitory effects on digestive enzymes of carnosic acid in sage and rosemary.

In this study, the aim was to determine the bioaccessibilities of carnosic acid in sage and rosemary and in vitro inhibitory effects of these samples on lipid and starch digestive enzymes by evaluating the lipase, α-amylase and α-glucosidase enzyme inhibition activities. The content of carnosic acid in rosemary (18.72 ±â€¯0.33 mg/g) was found to be higher than that content of that in sage (3.76 ±â€¯0.13 mg/g) (p < 0.05). The carnosic acid bioaccessibilities were found as 45.10 ±â€¯1.88% and 38.32 ±â€¯0.21% in sage and rosemary, respectively. The tested sage and rosemary showed inhibitory activity against α-glucosidase (Concentration of inhibitor required to produce a 50% inhibition of the initial rate of reaction - IC50 88.49 ±â€¯2.35, 76.80 ±â€¯1.68 µg/mL, respectively), α-amylase (IC50 107.65 ±â€¯12.64, 95.65 ±â€¯2.73 µg/mL, respectively) and lipase (IC50 6.20 ±â€¯0.63, 4.31 ±â€¯0.62 µg/mL, respectively). Furthermore, to the best of our knowledge, this is the first work that carnosic acid standard equivalent inhibition capacities (CAEIC50) for these food samples were determined and these values were in agreement with the IC50 values. These results show that sage and rosemary are potent inhibitors of lipase, α-amylase and α-glucosidase digestive enzymes.

In vitro biological assessment of Homalium zeylanicum and isolation of lucidenic acid A triterpenoid.

Homalium zeylanicum (Gardner) Benth. (Flacourtiaceae) is a medicinal plant useful in controlling rheumatism, inflammation and diabetes. The objective of this work evaluates in vitro antioxidant, antidiabetic, and antiinflammatory properties of hydroalcohol extract of bark of H. zeylanicum (HAHZ). It also describes isolation and structure determination of lucidenic acid A, which is the first report in this plant. In order to explain the role of antioxidant principles, DPPH, nitric oxide, hydroxyl, superoxide and metal chelating assays were performed. Antidiabetic and anti-inflammatory activities were investigated by quantifying α-amylase, α-glucosidase and protein denaturation inhibitory activities of HAHZ. Biochemical estimations were performed. The chemical structure of the triterpenoid was elucidated using 1H, 13C NMR and high resolution-MS. IC50 of DPPH, nitric oxide, hydroxyl, superoxide and metal chelating activities were of 36.23 ± 0.27, 40.11 ± 0.32, 35.23 ± 0.57, 43.34 ± 0.22 and 11.54 ± 0.08 µg/mL, respectively. IC50 of α-amylase and α-glucosidase activities were 29.12 ± 0.54, and 18.55 ± 0.15 µg/mL. Total phenolic and total flavonoid contents were recorded at 233.65 mg/g GAE and 172.7 mg/g QE. Regarding kinetic behaviour, HAHZ showed competitive inhibition on α-glucosidase and mixed competitive inhibition on α-amylase. Lucidenic acid A was confirmed by spectroscopic studies. Anti-inflammatory activity of lucidenic acid A was determined by using protein denaturation assay with IC50 13 µg/mL but HAHZ showed 30.34 ± 0.13 µg/mL. Phenols and flavonoids could be attributed to inhibition of intestinal carbohydrases for anti-diabetic activities whereas triterpenoids could be responsible for anti-inflammatory activity of H. zeylanicum.

Inhibitory effects of chickpea and Tribulus terrestris on lipase, α-amylase and α-glucosidase.

The total saponin content and its in vitro bioaccessibilities in Tribulus terrestris and chickpea were determined by a static in vitro digestion method (COST FA1005 Action INFOGEST). Also, in vitro inhibitory effects of the chosen food samples on lipid and starch digestive enzymes were determined by evaluating the lipase, α-amylase and α-glucosidase activities. The tested T. terrestris and chickpea showed inhibitory activity against α-glucosidase (IC50 6967 ± 343 and 2885 ± 85.4 µg/ml, respectively) and α-amylase (IC50 343 ± 26.2 and 167 ± 6.12 µg/ml, respectively). The inhibitory activities of T. terrestris and chickpea against lipase were 15.3 ± 2.03 and 9.74 ± 1.09 µg/ml, respectively. The present study provides the first evidence that these food samples (T. terrestris, chickpea) are potent inhibitors of key enzymes in digestion of carbohydrates and lipids in vitro.

Ethyl acetate fraction of Aframomum melegueta fruit ameliorates pancreatic ß-cell dysfunction and major diabetes-related parameters in a type 2 diabetes model of rats.

ETHNOPHARMACOLOGICAL RELEVANCE: In West Africa, various preparations of the fruit, seed and leaf of Aframomum melegueta K. Schum. are reputably used for the management of diabetes mellitus (DM) and other metabolic disorders. The present study evaluated the anti-diabetic effects of A. melegueta ethyl acetate fraction (AMEF) from fruit ethanolic extract in a type 2 diabetes (T2D) model of rats. MATERIALS AND METHODS: T2D was induced in rats by feeding a 10% fructose solution ad libitum for two weeks followed by a single intraperitoneal injection of streptozotocin (40 mg/kg body weight) and the animals were orally treated with 150 or 300 mg/kg body weight (bw) of the AMEF once daily for four weeks. RESULTS: At the end of the intervention, diabetic untreated animals showed significantly higher serum glucose, serum fructosamine, LDH, CK-MB, serum lipids, liver glycogen, insulin resistance (HOMA-IR), AI, CRI and lower serum insulin, pancreatic ß-cell function (HOMA- ß) and glucose tolerance ability compared to the normal animals. Histopathological examination of their pancreas revealed corresponding pathological changes in the islets and ß-cells. These alterations were reverted to near-normal after the treatment of AMEF at 150 and 300 mg/kg bw when, the effects were more pronounced at 300 mg/kg bw compared to the 150 mg/kg bw. CONCLUSION: The results of our study suggest that AMEF treatment at 300 mg/kg bw showed potent anti-diabetic effect in a T2D model of rats.

Partial Purification and Characterization of β-glucosidase from Monascus sanguineus

The aim of the present work was to study the production and characterization of β-glucosidase from Monascus sanguineus. Agro-waste residues were screened to obtain the maximum yield of enzyme. Jack fruit seed was the best substrate for enzyme production. Studies on the optimization of pH and temperature showed acidic pH favorable for enzymatic activity, whereas the optimum temperature was 60°C. Enzyme kinetics studies with different concentration of pNPG showed the calculated value of Km approximately 0.89 mM with the non-linear regression and 0.98 mM with the linear regression techniques. The enzyme was predominantly inhibited by KCl (69.8%) and moderately inhibited by CaCl2 (14.8%). Studies on the sensitivity for glucose showed that after 100 mM concentration of glucose, inhibition in pNPG hydrolysis took place. The molecular weight of the protein was estimated as 116 and 66 kDa with SDS- PAGE and zymography was carried out to verify the specific activity.

The psychology postgraduate system in Brazil: current characteristics and challenges for the area / O sistema de pós-graduação no Brasil: características atuais e desafios à área

AbstractThis text is a characterization of the Psychology Postgraduate System in Brazil, presenting indicators of its growth in recent years, of its current geographical distribution, of its research lines, and of the grades awarded in the last Triennial evaluation of 2013. This characterization underlies a set of reflections regarding the challenges for the area from the guidelines established by the National Postgraduate System Plan (PNPG) 2011-2020. These challenges include: internationalization, interdisciplinarity, the link with basic education, the reduction of regional disparities, and the differentiation between academic and professional qualification. The text also focuses on the continuous and necessary improvements in the evaluation process conducted by the Coordination for the Improvement of Higher Education Personnel (CAPES). These reflections seek to provide a basis for the management of the Psychology Postgraduate System in Brazil in the coming years. (AU)

ResumoO presente texto faz uma caracterização do sistema de Pós-Graduação em Psicologia no Brasil, oferecendo indicadores do seu crescimento nos últimos anos, da distribuição geográfica vigente, dos fenômenos tomados como objetos de investigação nas suas linhas de pesquisa e das notas atribuídas na última avaliação trienal de 2013. Tal caracterização embasa um conjunto de reflexões sobre os desafios para a Área a partir de diretrizes fixadas pelo Programa Nacional de Pós-Graduação (PNPG) 2011-2020, tais como: internacionalização, interdisciplinaridade, vínculo com a educação básica, redução das assimetrias regionais, diferenciação entre a formação acadêmica e profissional. Focaliza-se, também, os aprimoramentos contínuos e necessários do processo de avaliação conduzido pela Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Capes. Tais reflexões buscam embasar a gestão da Área nos próximos anos. (AU)

Butanol fraction of Khaya senegalensis root modulates ß-cell function and ameliorates diabetes-related biochemical parameters in a type 2 diabetes rat model.

ETHNOPHARMACOLOGICAL RELEVANCE: Khaya senegalensis A. Juss (Meliaceae) is commonly exploited for the traditional treatment of diabetes mellitus in Nigeria and Togo. The present study was conducted to examine the anti-diabetic activity of Khaya senegalensis butanol fraction (KSBF) of root ethanolic extract in a type 2 diabetes (T2D) model of rats. MATERIALS AND METHODS: T2D was induced in rats by feeding a 10% fructose solution ad libitum for two weeks followed by a single intraperitoneal injection of streptozotocin (40 mg/kg body weight) and the animals were treated with 150 and 300 mg/kg body weight (BW) of the fraction for five days in a week. Relevant diabetes-related parameters were analyzed in all experimental animals. RESULTS: The KSBF treatment, at 300 mg/kg BW, significantly (p<0.05) reduced blood glucose level, improved oral glucose tolerance ability and ß-cell function (HOMA-ß), decreased insulin resistance (HOMA-IR), stimulated hepatic glycogen synthesis, ameliorated serum lipids alterations and prevented hepatic and renal damages compared to untreated diabetic rats. Additionally, the fraction insignificantly (p>0.05) improved weight gain, decreased food and fluid intake, stimulated insulin secretion and lowered serum fructosamine concentrations compared to untreated diabetic rats. CONCLUSIONS: Data from this study suggests that orally administered KSBF, at 300 mg/kg BW, possess remarkable anti-type 2 diabetic activity and could ameliorate some diabetes-associated complications and hence can be considered as a source of potential anti-type 2 diabetic medicine.

Anti-diabetic effects of the acetone fraction of Senna singueana stem bark in a type 2 diabetes rat model.

ETHNOPHARMACOLOGICAL RELEVANCE: Senna singueana is currently used in the traditional treatment of diabetes mellitus in Nigeria. The present study examined the anti-diabetic activity of the Senna singueana acetone fraction (SSAF) of stem bark in a type 2 diabetes (T2D) rat model. MATERIALS AND METHODS: Crude ethyl acetate extract of the Senna singueana stem bark was fractionated with various solvents and the acetone fraction was selected for in vivo studies based on the high α-glucosidase and α-amylase inhibitory activities. In the in vivo study, male Sprague-Dawley rats were induced with T2D and treated with the SSAF at 150 and 300 mg/kg body weight. Several T2D-related parameters were measured in the study. RESULTS: After 4 weeks of intervention, non-fasting blood glucose concentrations were significantly decreased and the glucose tolerance ability was significantly improved in the SSAF treated groups compared to the diabetic control group. Serum insulin concentrations, pancreatic ß-cell function (HOMA-ß) and liver glycogen were significantly (P<0.05) increased while serum alanine transaminase, alkaline phosphatase and urea were significantly decreased in the SSAF treated diabetic rats compared to the diabetic control group. Though insignificantly (P>0.05), other T2D-induced abnormalities such as food and fluid intake, body weight, serum lipids, serum fructosamine level and peripheral insulin resistance (HOMA-IR) were also partially ameliorated by the SSAF treatment. CONCLUSION: Data of this study suggest that orally administered SSAF could ameliorate most of the T2D-induced abnormalities in a T2D model of rats.

Fast detection and quantification of Escherichia coli using the base principle of the microbial fuel cell.

Escherichia coli is an important microbial indicator of fecal contamination, making accurate quantitative detection of E. coli a key to ensuring public health. In this study, a microbial fuel cell (MFC) was used as a detection unit of an E. coli sensor, and specific enzymes expressed in E. coli, such as ß-D-galactosidase (GAL) and ß-D-glucuronidase (GUS), were exploited as biological detection elements. As substrates, 4-aminophenyl-ß-D-galactopyranoside (4-APGal) were used for GAL detection, whereas 8-hydroxyquinoline glucuronide (8-HQG) and 4-nitrophenyl ß-D-glucuronide (PNPG) were used for GUS detection. Once these substrates were hydrolyzed by GAL or GUS, they became electrochemically active products, which were, in turn, oxidized on the anode of the MFC reactor. The power output of the MFC reactor increased sharply when E. coli in the reactor reached the critical concentration. Accordingly, the time required to reach the highest voltage output was recorded as a detection time (DT), and a negative linear relationship was established between DT and the logarithm of the initial concentration of E. coli in the samples studied. The DTs of laboratory samples were 140 min and 560 min for initial concentrations of 1.9 × 10(7) CFU/mL and 42 CFU/mL at 44.5 °C. Moreover, the DTs for GUS assays were further shortened by induction with methyl ß-D-glucuronide sodium salt (MetGlu). The quantitative relationship between DTs and initial E. coli concentrations established from replicate laboratory sample assays allowed estimation of the E. coli concentration in environmental samples, but with approximately 100 min of lag time. The lag time was also observed with E. coli samples that were prepared by starving cells in a laboratory.

Inhibition of pancreatic lipase, α-glucosidase, α-amylase, and hypolipidemic effects of the total flavonoids from Nelumbo nucifera leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: Nelumbo nucifera Gaertn. leaves have been used as medicinal herbs in the past 1300 years, specifically utilized to cure hyperlipidemia, hyperglycemia, and obesity. It has been recorded in the most famous medicinal book in China for more than 400 years. The present study aims to identify the potential therapeutic activities of the flavonoids isolated from Nelumbo nucifera leaves. MATERIALS AND METHODS: Nelumbo nucifera leaf flavonoids (NLF) were tested for the inhibition of lipase, α-glucosidase, and α-amylase activities in vitro. A single dose of NLF was administered by oral gavage in mice for acute toxicity. Wistar rats with high-fat diet-induced hyperlipidemia and two other animal models were used to evaluate the hypolipidemic effects of NLF. RESULTS: Our in vitro biochemistry tests revealed that the NLF showed high inhibitory activity against porcine pancreatic lipase, α-amylase, and α-glucosidase with IC50 values of 0.38 ± 0.022, 2.20 ± 0.18, and 1.86 ± 0.018 mg/mL, respectively. Furthermore, the NLF significantly lowered the lipid components, such as the total cholesterol, triglycerides, low-density lipoprotein cholesterol, and malondialdehyde, in various established in vivo systems and raised the high-density lipoprotein cholesterol. Moreover, the NLF alleviated high-fat diet-induced lipid accumulation in the liver. CONCLUSIONS: The results demonstrate that NLFs can effectively ameliorate hyperlipidemia and inhibit the key enzymes related to type 2 diabetes mellitus. Our findings may provide new pharmacological basis for the treatment of hyperlipidemia, hyperglycemia, and obesity using NLFs.