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This study was designed to screen 6 lysosomal storage diseases (LSDs) in neonates using tandem mass spectrometry (MS/MS), and establish cutoff values for these LSDs with 3000 dried blood spots (DBS) samples. Cutoff values for α-L-iduronidase (IDUA), α-galactosidase (GLA), acid beta glucosidase (ABG), β-galactocerebrosidase (GALC), acid sphingomyelinase (ASM), and acid alpha glucosidase (GAA) were as follows: GLA, > 2.06 μmol/L·h; ABG, > 1.78 μmol/L·h; ASM, > 0.99 μmol/L·h; IDUA, > 1.33 μmol/L·h; GALC, > 0.84 μmol/L·h; and GAA, > 2.06 μmol/L·h. There were 30 positives in initial MS/MS screening test, and 15 samples were still positive with repeat testing. Their parents/guardians were recontacted and DBS samples were collected again for test. Only 1 child showed abnormal GAA enzyme activity after recontacting process, and was diagnosed with Pompe disease after genetic screening. Eventually, cutoff values of 6 specific enzyme activities were established and MS/MS is effective for early LSDs screening., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Nitraria roborowskii Kom (NRK), with high economic and ecological value, is mainly distributed in the Qaidam Basin, China. However, research on its chemical components and bioactivities is still rare. In this study, its chemical constituents (52) including 10 β-carboline alkaloids, nine cyclic peptides, three indole alkaloids, five pyrrole alkaloids, eight phenolic acids and 17 flavonoids were identified tentatively using UPLC-triple-TOF-MS/MS. Notablely, one new β-carboline alkaloid and five new cyclic peptides were confirmed using MS/MS fragmentation pathways. In addition, experiments in vitro indicated that NRK-C had strong maltase and sucrase inhibitory activities (IC 50 of 0.202 and 0.103 mg/mL, respectively). Polysaccharide tolerance experiments confirmed NRK-C (400 mg/kg) was associated with decreased postprandial blood glucose (PBG) in diabetic mice. These results suggested that NRK fruit might be used as a functional ingredient in food products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

As one of the most important industrial enzymes, α-amylase is widely used in food processing, such as starch sugar and fermentation, bringing high added value to industry of more than a trillion dollars. We developed a multi-enzyme system (Glu&Gox@Cu-MOF-74) prepared by embedding α-glucosidase (Glu) and glucose oxidase (Gox) into the biomimetic metal-organic framework Cu-MOF-74 using in situ encapsulation within 15 min at room temperature for efficient and sensitive detection of α-amylase activity. Benefitting from the remarkable peroxidase-mimicking property and rigid skeleton of Cu-MOF-74, the biocatalytic platform exhibited excellent cascade activity and tolerance in various extremely harsh environments compared to natural enzymes. On this basis, a cascade biocatalytic platform was constructed for the detection of α-amylase activity with wide linear range (5-100 U/L) and low limit of detection (1.45 U/L). The colorimetric cascade scheme is important for the sensitive and selective determination of α-amylase in complex fermentation samples, and the detection time is short (∼0.5 h). This work provides new ideas for the detection of α-amylase based on the cascade amplification method., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Starch-based dietary fibers are at the forefront of functional ingredient research. In this study, a novel water-soluble slow digestion dextrin (SDD) was synthesized by synergy of α-glucosidase and cyclodextrin glucosyltransferase and characterized. Results showed that SDD exhibited high solubility, low viscosity, and resistance to digestive enzymes, and also showed an increased dietary fiber content of 45.7% compared with that of α-glucosidase catalysis alone. Furthermore, SDD was used as the sole carbon source to ferment selected intestinal strains and human fecal microflora in vitro to investigate its prebiotic effects. It was found that SDD could markedly enriched the abundance of Bifidobacterium, Veillonella, Dialister, and Blautia in human gut microflora and yielded higher total organic acid. The combination of α-glucosidase and cyclodextrin glucosyltransferase in this study showed valuable potential for the preparation of a novel slow digestion dextrin with good physicochemical properties and improved prebiotic effects., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Background : Berry fruits are recognized as a "superfood" due to their high content of bioactive compounds and health benefits. Scope and approach : Herein, extracts of Cornus sanguinea and Cornus mas fresh and dried fruits obtained by different extraction procedures (ethanolic and hydroalcoholic maceration, ultrasound-assisted extraction, and Soxhlet apparatus) were analysed using liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (LC-ESI-QTOF-MS) and compared to identify the main healthy compounds and their impact on the inhibition of key enzymes (pancreatic lipase, α-glucosidase, and α-amylase) associated with metabolic disorders. The antioxidant activity and inhibition of nitric oxide (NO) and NF-κB pathway were also investigated. Key findings and conclusions : Flavonoids, iridoids, and phenolic acids were the main classes of identified compounds. Herein, kaempferol 3- O -galactoside, kaempferol 3- O -glucoside, quercetin, quercetin 3- O -xyloside, and myricetin 3- O -galactoside were detected for the first time in C. sanguinea . Remarkable antioxidant effects and promising α-glucosidase and lipase inhibitory activity were observed with extracts obtained by hydroalcoholic maceration of both Cornus dried fruits. Consequently, these extracts were subjected to fractionation using Amberlite XAD-16 resin. The most promising biological activities, which are attributed to the presence of some flavonoids and iridoids, were detected with the C. sanguinea fractions, in particular SD2(II). The results of this study offer new insights into the potential development of functional foods, nutraceuticals, and food supplements using the Cornus species.

A total of nine new phenolic amides ( 1 - 9 ), including four pairs of enantiomeric mixtures ( 3 - 5 and 8 ), along with ten known analogues ( 10 - 19 ) were identified from the fruits of Lycium barbarum using bioassay-guided chromatographic fractionation. Their structures were elucidated by comprehensive spectroscopic and spectrometric analyses, chiral HPLC analyses, and quantum NMR, and electronic circular dichroism calculations. Compounds 5 - 7 are the first example of feruloyl tyramine dimers fused through a cyclobutane ring. The activity results indicated that compounds 1 , 11 , and 13 - 17 exhibited remarkable inhibition against α-glucosidase with IC 50 of 1.11-33.53 μM, 5-150 times stronger than acarbose (IC 50 = 169.78 μM). Meanwhile, compounds 4a , 4b , 5a , 5b , 13 , and 14 exerted moderate agonistic activities for peroxisome proliferator-activated receptor (PPAR-γ), with EC 50 values of 10.09-44.26 μM. Especially,compound 14 also presented inhibitory activity on dipeptidyl peptidase-4 (DPPIV), with an IC 50 value of 47.13 μM. Furthermore, the banding manner of compounds 14 and 17 with the active site of α-glucosidase, DPPIV, and PPAR-γ was explored by employing molecular docking analysis.

Incidents and accidents often involve the drinking of alcoholic beverages. We investigated compounds that indicate the consumption of alcoholic beverages even after ethanol (EtOH) becomes undetectable in blood and urine. Ethyl glucoside (EG) has been isolated as a possible drinking marker, and a GC-MS/MS method for EG isomers has been developed. EG isomers in several alcoholic beverages were analyzed. In sake, only αEG was observed in high concentrations. In wine and beer, both α and βEG were detected. Whisky, however, did not contain EG. EtOH and EG concentrations were analyzed in urine up to 48 h after ingestion. Maximum EtOH concentrations were reached in 1-2 h and was mostly eliminated in 6 h. Maximum EG concentrations were reached in 3-6 h, gradually decreased, and remained low after 24 h. After drinking sake, the αEG concentrations were much higher than that of other alcoholic beverages. After drinking wine or beer, βEG was detected, but lower than αEG. Also, αEG was detected in urine after drinking whisky that contained no EG. This suggested that αEG may be synthesized in vivo. Disaccharide-degrading enzymes such as α-glucosidase are present in the human small intestine. It was considered that αEG was synthesized when alcohol was consumed with certain foods, such as carbohydrates. In actual forensic autopsy cases, EtOH and EG isomer analysis provided useful information regarding drinking history. In conclusion, it is considered that urinary EG isomers can be used as drinking markers that complement EtOH analysis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

In this work, the volatile fingerprints of different parts of Chongming saffron flowers (stigmas, stamens, and tepals) were analyzed and compared for the first time by headspace-gas chromatography-ion mobility spectrometry. Three different parts of saffron flowers could be clearly distinguished using principal component analysis based on signal intensity data of gas chromatography-ion mobility spectrometry. Therefore, gas chromatography-ion mobility spectrometry coupled with principal component analysis method could be employed as a new method for authentication and quality control of saffron for the reason of frequent addition with stamens and/or tepals as adulterants in saffron. Moreover, the bioactive composition (total flavonoids, total phenolics, and total anthocyanins) and bioactive properties of saffron tepals were evaluated. The results indicated that aqueous, ethanol, and ethyl acetate extracts of saffron tepals exhibited good radical scavenging (2,2-Diphenyl-1-picrylhydrazyl, ABTS, and OH) and enzyme (α-amylase/α-glucosidase) inhibition activities, which probably were attributed to the bioactive components contained in the extracts. This approach would provide the important information for monitoring the quality of saffron as well as exploring the utilization of saffron tepals in functional food technology. PRACTICAL APPLICATION: This study demonstrated that the HS-GC-IMS method might be used as a new strategy for quality control of saffron, and the saffron tepals were rich source of bioactive components that could be used in health-promoting products., (© 2022 Institute of Food Technologists®.)

Chinese Tibetan tea made from the tender leaves of Malus transitoria is a widely consumed health drink, but there are few reports on its chemical composition and biological activity. In this study, we found that a 50% ethanol extract of M. transitoria had good antioxidant and α-glucosidase inhibitory activities in vitro. Guided by in vitro bioassays, chromatographic separation and purification were conducted, and the most active fraction in M. transitoria was determined. UPLC-Orbitrap-MS/MS was used to further quickly and comprehensively characterize the chemical composition. Library searches, MS/MS fragmentation patterns of two isolated reference compounds, and bibliography were used to annotate 81 compounds, of which 2 were new compounds, and 79 were identified from M. transitoria for the first time. This study provides a scientific basis for the development of antioxidant and anti-diabetic functional foods from M. transitoria., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

A highly stable and reusable fluorescent multisample nanobiosensor for the detection of α-glucosidase inhibitors has been developed by coupling fluorescent liposomal nanoparticles based on conjugated polymers (L-CPNs) to the enzyme α-glucosidase, one of the main target enzymes in the treatment of type 2 diabetes. The mechanism of sensing is based on the fluorescence "turn-on" of L-CPNs by p -nitrophenol (PNP), the end product of the enzymatic hydrolysis of p -nitrophenyl-α-d-glucopyranoside. L-CPNs, composed of lipid vesicles coated with a blue-emitting cationic polyfluorene, were designed and characterized to obtain a good response to PNP. Two nanobiosensor configurations were developed in this study. In the first step, a single-sample nanobiosensor composed of L-CPNs and α-glucosidase entrapped in a sol-gel glass was developed in order to characterize and optimize the device. In the second part, the nanobiosensor was integrated and adapted to a multiwell microplate and the possibility of reusing it and performing multiple measurements simultaneously with samples containing different α-glucosidase inhibitors was investigated. Using super-resolution confocal microscopy, L-CPNs could be visualized within the sol-gel matrix, and the quenching of their fluorescence, induced by the substrate, was directly observed in situ . The device was also shown to be useful not only as a platform for screening of antidiabetic drugs but also for quantifying their presence. The latter application was successfully tested with the currently available drug, acarbose.

Objective: To provide the scientific basis for the utility of rhizome of Trillium govanianum as nutraceutical supplements in managing physiological glycemic levels., Methods: The in vitro enzyme inhibitory activity of the extract, fractions, and the isolated steroidal saponins from the rhizome part of T. govanianum was carried out against α-amylase, α-glucosidase, and dipeptidyl peptidase IV. The molecular interactions, binding score, and pharmacokinetic parameters (absorption, distribution metabolism, and excretion) of steroidal saponins were analyzed by the Schrodinger molecular docking software., Key Findings: Current study explained that the extract, fractions, and isolated steroidal saponins from T. govanianum possess good α-amylase and α-glucosidase inhibitory activity while moderate dipeptidyl peptidase IV inhibitory activity. Moreover, in vitro results revealed that borassoside E (IC50 7.15 ± 1.78 μM), protodioscin (IC50 6.72 ± 0.04 μM), and diosgenin (IC50 12.75 ± 2.70 μM) are most effective in inhibiting the activity of α-amylase, α-glucosidase, and dipeptidyl peptidase IV, respectively. Current in silico and in vitro studies established an association between the steroidal saponins from T. govanianum and their molecular interactions with α-amylase, α-glucosidase, and dipeptidyl peptidase IV., Conclusion: The results of this investigation suggest that fractions and steroidal saponins from T. govanianum exhibit good antidiabetic activity which could be used as nutraceutical supplements for the management of systemic glucose level., (© The Author(s) 2021. Published by Oxford University Press on behalf of Royal Pharmaceutical Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

We investigated the effects of supplementation of yogurt with elderberry juice (Sambucus nigra L.), in both natural and restructured forms, on certain technological, physicochemical, sensory, and health-promoting properties of yogurt, including antioxidant activity and inhibition of angiotensin-converting enzyme (ACE), α-amylase, and α-glucosidase activity. Consumer acceptance of the yogurt-juice products was assessed. Gel-strength restructured elderberry juice retained a spherical shape and most of the juice, despite decreasing in mass from 57.2 to 50.9 g during storage. As a result, yogurt supplemented with 10 and 25% restructured elderberry juice appeared to be more desirable from a sensory and technological perspective than yogurt with natural juice. Yogurt supplemented with restructured elderberry juice had a high water-holding capacity (94.4-96.4%), exhibited no spontaneous whey syneresis, and maintained a dense consistency (up to 5,626 g). Consumer penalty analysis of the just-about-right diagnostic attributes indicated that the flavor of these yogurts may not be sufficiently refreshing. High correlation was demonstrated between ACE inhibition, ABTS, α-amylase, and α-glucosidase in yogurt supplemented with restructured juice. An in vitro gastrointestinal simulation estimated bioaccessibility of antioxidants to be in the range of 62 to 66%. This model fermented yogurt supplemented with restructured elderberry juice is a novel dairy-juice beverage that represents a new approach for the development of functional fruit yogurt beverages., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The high incidence of diabetes mellitus has caused widespread concern around the world, and has quickly become one of the most prevalent and costly chronic diseases. Alpha-glucosidase has been considered as the main target enzyme in the prevention and treatment of type 2 diabetes, and the inhibition of alpha-glucosidase activity can control postprandial blood glucose levels of diabetics and keep blood glucose levels normal. However, most of the current antidiabetic drugs in the clinic have many side effects. Recently, the screening of alpha-glucosidase inhibitors, from natural products, with excellent activity and less side effects has been successfully achieved, which also proposes more requirements and challenges for the approaches of alpha-glucosidase inhibitor (AGI) screening because of the trace level of active compounds in natural products, large-scale sample screening, and complex matrix of natural products. In this review, we comprehensively summarized the available strategies for the assay of alpha-glucosidase activity and AGI screening for the first time, including classic strategies such as high throughput screening methods (e.g. capillary electrophoresis strategies, HPLC/LC-MS strategies), emerging sensing strategies (e.g. colorimetric sensing, fluorescence sensing, electrochemical sensing, and surface plasmon resonance sensing) and other strategies. Finally, future opportunities and challenges in future research are tentatively proposed.

In recent years, α-glucosidase (α-Glu) inhibitor has been widely used in clinic for diabetic and HIV therapy. Although different systems have been constructed for sensitive and selective detection of α-Glu and screening its inhibitor, the method based on ratiometric fluorescence for α-glucosidase inhibitor screening remains poorly investigated. Herein, we constructed a new MnO 2 nanosheet (NS)-based ratiometric fluorescent sensor for α-glucosidase activity assay and its inhibitor screening. MnO 2 NS as an oxidase-mimicking nanomaterial directly oxidized o-phenylenediamine (OPD) into 2,3-diaminophenazine (DAP) which had a strong fluorescence emission at 575 nm, whereas the fluorescence of Ag nanoclusters (NCs) at 450 nm was then quenched by the generated DAP through inner filter effect (IFE). When 2-O-α-d-glucopyranosyl-l-ascorbic acid (AAG) as α-Glu substrate and α-Glu were introduced into the above system, MnO 2 NS would be reduced to Mn 2+ and lose the oxidase-like property since ascorbic acids (AA) were released with the hydrolysis of AAG by α-Glu. Thus, DAP would not be produced and IFE was stopped accompanying with the fluorescence decrease of DAP and fluorescence increase of AgNCs. A ratiometric fluorescent α-Glu nanosensor was thus developed. The fluorescence intensity ratio of DAP to AgNCs linearly decreased with the increasing of α-Glu concentrations in the range of 0.2-8 U mL -1 , and limit of detection was 0.03 U mL -1 . This proposed sensing approach was also expanded to α-Glu inhibitor screening and showed excellent applicability. As a typical α-Glu inhibitor, acarbose was investigated with a low detection limit of 10 -8  M. The constructed sensor platform was proven to be sensitive and selective as well as simple, label-free and low-cost, making it promising for the accurate diagnosis of relevant disease and discovery of potential drugs., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Microsurgical vasoepididymostomy (MVE) is recommended as a first-line option for treatment of epididymal obstructive azoospermia (EOA). However, early indicators for predicting patency and natural pregnancy are unclear. Our aim was to explore the early predictive value of seminal plasma neutral alpha-glucosidase (NAG) activity for patency and natural pregnancy after MVE. Eighty-four patients with EOA who underwent MVE were enrolled in this study. The post-operative patency and natural pregnancy rates were 60.71% and 33.33% respectively. The presence of motile epididymal spermatozoa at the anastomosis site and NAG activity measured at the first month after MVE were early and independent predictors of patency and natural pregnancy. The areas under the receiver operating characteristic curves (AUCs) of NAG activity for prediction of patency and natural pregnancy were 0.78 (95% confidence interval [CI]: 0.68-0.88) and 0.82 (95% CI: 0.73-0.92). The best cut-off values of NAG activity for predicting patency and pregnancy were 15.9 and 17.0 m IU/ejaculate respectively. In conclusion， NAG activity measured at the first month after MVE is an early and independent predictor of patency and natural pregnancy., (© 2019 Blackwell Verlag GmbH.)

Various studies have sought to determine the typical v. optimal abstinence period after which semen samples should be collected, with many contradictory results reported. Several factors influence the semen microenvironment, and thus sperm parameters. In this study we focused on the secretions of the prostate, seminal vesicles and the epididymis. Semen samples were obtained from healthy normozoospermic males (n=16) after 4-day and 4-h periods of ejaculatory abstinence, and standard semen analysis was performed using computer-aided sperm analysis, whereas seminal plasma citric acid, neutral α-glucosidase and fructose concentrations were measured using assay kits. There were significant decreases in total sperm count (P<0.001), sperm concentration (P<0.05) and semen volume (P<0.05) after 4h compared with 4 days ejaculatory abstinence. Furthermore, increases were observed in total sperm motility (P<0.05) and sperm progressive motility (P<0.01) after a 4-h abstinence period, accompanied by significant reductions in citric acid (P<0.05), α-glucosidase (P<0.01) and fructose (P<0.01) concentrations. In addition, due to the decreased number of spermatozoa, these concentrations translated to a significant decrease in fructose (P<0.05) per spermatozoon, indicating an intrinsic mechanism capitalising on alternative sources of energy for increased metabolic function and subsequent sperm motility.

Molecules designed for cell-specific imaging were studied, taking advantage of an enzyme-inhibitor interaction. 1-Deoxynojirimycin (DNJ) can be actively captured by cells which express the surface membrane protein α-glucosidase. New probes composed of DNJ for recognition linked to a fluorophore signal portion were prepared (DNJ-CF 3 1, DNJ-Dans 2 and DNJ-DEAC 3). Docking simulations revealed that the inhibitors acarbose and miglitol and the inhibitor portion of the probes bind at the same position in the pocket of α-glucosidase (human-derived PDB: 3TON). The ability of probes 1-3 to detect the difference between HeLa cells (from human cervical cancer tissue), Neuro-2a cells (from a mouse neuroblastoma C1300 tumor), N1E-115 cells (from a mouse brain neuroblastoma C1300 tumor), A1 cells (from the astrocyte of a newborn mouse brain), and Caco-2 cells (from a human colon carcinoma) was evaluated, and cell-specific fluorescence imaging was possible for conjugate probes 1 and 2. Caco-2 cells treated with probes 1 and 2 showed blue and green fluorescence, respectively, from the cell membrane, and did not stain the Caco-2 cells inside. These results show that DNJ-CF 3 1 and DNJ-Dans 2 recognize an α-glucosidase protein on the surface of Caco-2 cells. Probes 1 and 2 did not stain any part of the other cells. This cell-specific imaging strategy is applicable for a variety of therapeutic agents for many diseases., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

A reusable biosensor has been fabricated in this work for the assay of α-glucosidase activity and the inhibitor screening. In this design, the aptamer of ATP is split as split aptamer 1 (Apt 1) and split aptamer 2 (Apt 2), and Apt 2 can link gold nanoparticles (AuNPs) modified with Apt 1 and 4-aminophenyl-α-d-glucopyranoside (pAPG). Consequently, the functional AuNPs can be immobilized onto the surface of gold electrode, allowing for salt-induced regeneration. In the presence of α-glucosidase, the glycosyl of pAPG is cut off, and the electroactive phenolic hydroxyls appear to give a strong current signal. Furthermore, the biosensor can be recovered very easily by incubating it in water to dissociate the AuNPs modified with Apt 1 and pAPG. So, a new biosensor for α-glucosidase activity detection and inhibitor screening is developed based on enzyme-activated signal generation and recovery. The biosensor may also exhibit good sensitivity for α-glucosidase determination with the detection limit 0.005 U/mL and can be reused by water-washing regeneration with good repeatability. Meanwhile this biosensor can also be utilized for inhibitor screening, which may have potential for clinical applications., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Seasonal variations in crops can alter the profile and amount of constituent compounds and consequentially any biological activity. Differences in phytochemical profile, total phenolic content and inhibitory activity on α-glucosidase (maltase) of Hibiscus sabdariffa calyces grown in South Western Nigeria were determined over wet and dry seasons. The phenolic profile, organic acids and sugars were analysed using HPLC, while inhibition of rat intestinal maltase was measured enzymically. There was a significant increase (1.4-fold; p ≤ 0.05) in total anthocyanin content in the dry compared to wet planting seasons, and maltase inhibition from the dry season was slightly more potent (1.15-fold, p ≤ 0.05). Fructose (1.8-fold), glucose (1.8-fold) and malic acid (3.7-fold) were significantly higher (p ≤ 0.05) but citric acid was lower (62-fold, p ≤ 0.008) in the dry season. Environmental conditions provoke metabolic responses in Hibiscus sabdariffa affecting constituent phytochemicals and nutritional value., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Purpose: To describe a novel biochemical marker in dried blood spots suitable to improve the specificity of newborn screening for Pompe disease., Methods: The new marker is a ratio calculated between the creatine/creatinine (Cre/Crn) ratio as the numerator and the activity of acid α-glucosidase (GAA) as the denominator. Using Collaborative Laboratory Integrated Reports (CLIR), the new marker was incorporated in a dual scatter plot that can achieve almost complete segregation between Pompe disease and false-positive cases., Results: The (Cre/Crn)/GAA ratio was measured in residual dried blood spots of five Pompe cases and was found to be elevated (range 4.41-13.26; 99%ile of neonatal controls: 1.10). Verification was by analysis of 39 blinded specimens that included 10 controls, 24 samples with a definitive classification (16 Pompe, 8 false positives), and 5 with genotypes of uncertain significance. The CLIR tool showed 100% concordance of classification for the 24 known cases. Of the remaining five cases, three p.V222M homozygotes, a benign variant, were classified by CLIR as false positives; two with genotypes of unknown significance, one likely informative, were categorized as Pompe disease., Conclusion: The CLIR tool inclusive of the new ratio could have prevented at least 12 of 13 (92%) false-positive outcomes.

The mucosal maltase enzymes are characterized by an activity that produces glucose from linear glucose polymers, assayed with the disaccharide maltose. The related enzyme isomaltase produces glucose from branched glucose polymers, assayed with palatinose. Maltase and isomaltase activities are part of the 4 disaccharidases assayed from clinical duodenal biopsy homogenates. The reported maltase activities are more difficult to interpret than lactase or sucrase activities because both the sucrase-isomaltase and maltase-glucoamylase proteins have overlapping maltase activities. The early work of Dahlqvist identified 4 maltase activities from human small intestinal mucosa. On one peptide, sucrase (maltase Ib) and isomaltase (maltase Ia) activities shared maltase activities but identified the enzymes as sucrase-isomaltase. On the other peptide, no distinguishing characteristics of the 2 maltase activities (maltases II and III) were detected and the activities identified as maltase-glucoamylase. The nutritional/clinical importance of small intestinal maltase and isomaltase activities are due to their crucial role in the digestion of food starches to absorbable free glucose. This review focuses on the interpretation of biopsy maltase activities in the context of reported lactase, sucrase, maltase, and palatinase biopsy assay activity patterns. We present a classification of mucosal maltase deficiencies and novel primary maltase deficiency (Ib, II, III) and provide a clarification of the role of maltase activity assayed from clinically obtained duodenal biopsies, as a path toward future clinical and molecular genomic investigations.

Membrane-bound enzymes (MBEs), which make up a very high proportion of intracellular enzymes, catalyze a variety of activities that are currently analyzed by various techniques after purification. However, due to their amphipathic character, the purification of MBEs is difficult. Therefore, the most productive approach represents in situ analysis of MBEs in the cellular membrane. Methods: In this study, using membrane-bound α-glucosidase (α-Glu) as an example, we have developed a colorimetric in situ assay for MBEs based on the inhibitory effect of lipid bilayer on ion transport. The enzyme substrate could mediate the self-assembly of phospholipid PEG derivative around magnetic nanospheres that were modified with boronic acid. The formation of lipid bilayer could inhibit the leaking of iron ions under acidic conditions. However, the product of the catalytic reaction had no capability for self-assembly of the lipid bilayer, leading to the release of iron ions from the magnetic nanospheres under acidic pH. Results: The colorimetric in situ assay for MBEs could not only analyze the activity of membrane-bound α-Glu located on Caco-2 cells but could also evaluate the α-Glu inhibitors in cell medium. Conclusions: The simple, economic, and efficient method proposed here offers a potential application for high-throughput testing of α-Glu and its inhibitors. Our study also outlines a strategy for exploring the colorimetric method to detect the activities of MBEs in situ ., Competing Interests: Competing Interests: The authors have declared that no competing interest exists.

Human seminal plasma is rich in potential biological markers for male infertility and male reproductive system diseases, which have an application value in the diagnosis and treatment of male infertility. The methods for the detection of semen biochemical markers have been developed from the manual, semi-automatic to the present automatic means. The automatic detection of semen biochemical markers is known for its advantages of simple reagent composition and small amount of reagents for each test, simple setting of parameters, whole automatic procedure with few errors, short detection time contributive to batch detection and reduction of manpower cost, simple calibration and quality control procedure to ensure accurate and reliable results, output of results in the order of the samples in favor of clinical diagnosis and treatment, and open reagents applicable to various automatic biochemistry analyzers. At present, the automatic method is applied in the detection of such semen biochemical markers as seminal plasma total and neutral alpha-glucosidase, acid phosphatase, fructose, γ-glutamyl transpeptidase, zinc, citric acid, uric acid, superoxide dismutase and carnitine, sperm acrosin and lactate dehydrogenase C4, and semen free elastase, which can be used to evaluate the secretory functions of the epididymis, seminal vesicle and prostate, sperm acrosome and energy metabolism function, seminal plasma antioxidative function, and infection or silent infection in the male genital tract.

Pompe disease is a metabolic myopathy with a wide spectrum of clinical presentation. The gold-standard diagnostic test is acid alpha-glucosidase assay on skin fibroblasts, muscle or blood. Identification of two GAA pathogenic variants in-trans is confirmatory. Optimal effectiveness of enzyme replacement therapy hinges on early diagnosis, which is challenging in late-onset form of the disease due to non-specific presentation. Next-generation sequencing-based panels effectively facilitate diagnosis, but the sensitivity of whole-exome sequencing (WES) in detecting pathogenic GAA variants remains unknown. We analyzed WES data from 93 patients with confirmed Pompe disease and GAA genotypes based on PCR/Sanger sequencing. After ensuring that the common intronic variant c.-32-13T>G is not filtered out, whole-exome sequencing identified both GAA pathogenic variants in 77/93 (83%) patients. However, one variant was missed in 14/93 (15%), and both variants were missed in 2/93 (2%). One complex indel leading to a severe phenotype was incorrectly called a nonsynonymous substitution c.-32-13T>C due to misalignment. These results demonstrate that WES may fail to diagnose Pompe disease. Clinicians need to be aware of limitations of WES, and consider tests specific to Pompe disease when WES does not provide a diagnosis in patients with proximal myopathy, progressive respiratory failure or other subtle symptoms., (Copyright © 2017 Elsevier Inc. All rights reserved.)

This study investigated the effect of germination and fermentation on the composition of carbohydrates in Australian sweet lupin. Specifically, the amount of sugars (sucrose, fructose, and glucose), starch, oligosaccharides (verbascose, stachyose, and raffinose), and dietary fiber were measured in germinated lupin seeds and fermented lupin flour, and compared with those in soy. High performance liquid chromatography coupled with refractive index was employed for quantitation of sugars, starch, and oligosaccharides, and gas chromatography coupled with a flame ionization detector was used for quantitation of simple sugars in total, and soluble, and insoluble dietary fiber. The enzyme activities of α-amylase and α-glucosidase were compared before and after germination or fermentation. The α-amylase activity in germinated lupin increased to  ∼17 nmol/mL/min/0.1 g and in germinated soy∼32; in fermented lupin, the activity increased to ∼52, while in fermented soy it decreased to ∼20. In general, germination or fermentation decreased the oligosaccharide content, and increased the total sugar in samples (p < 0.05). Total oligosaccharides in lupin after uncontrolled germination were reduced by 98% to 6 mg/g, and after controlled germination reduced by 44% to 86 mg/g. Fermentation with yogurt culture lowered the content of total oligosaccharides due to 94% decrease in stachyose. Total oligosaccharides in soy flour prior to fermentation were 180 mg/g and significantly decreased to ∼124 mg/g in fermented soy. Germination did not affect the starch content. There was no significant change in the amounts of total, soluble, and insoluble dietary fiber after germination or fermentation of lupin except for galactose, which was significantly reduced in germinated lupin seeds. Soluble dietary fiber in germinated soy significantly increased. Germination and fermentation are simple and effective techniques to reduce the oligosaccharides while maintaining the composition of dietary fibers.

YingDe black tea is produced from crude tea prepared from leaves of Camellia sinensis var. assamica. In this work, we isolated and identified five novel flavanones, namely, amelliaone A-E (1-5), along with seven known compounds 6-12 from the ethanol extract of YingDe black tea. The structures of these five novel phenolic compounds were determined using extensive 1D and 2D nuclear magnetic resonance spectroscopy experiments. The compounds were further evaluated for antioxidant, α-glucosidase inhibitory, and cytotoxic activities. Compound 1 exhibited higher α-glucosidase inhibitory activity with a half-maximum inhibitory concentration value (IC 50 ) of 10.2µM compared with acarbose (18.2µM)., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Despite the wide use of extra virgin olive oil (EVOO) to combat several diseases, the antidiabetic and anti-cholinesterase activity of Spanish EVOO have not been assessed. In order to evaluate which compounds are responsible for these activities of five Spanish EVOOs, in addition to flavonoids, we investigated for the first time the effect of the contents of carotenoids, fatty acids (FAs), and phytoprostanes (PhytoPs) on four enzymes: α-glucosidase, α-amylase, acetylcholinesterase, and butyrylcholinesterase. The extracts of these five Spanish EVOOs were found to contain three flavones, three carotenoids, six FAs, and seven classes of PhytoPs. The samples exhibited no in vitro anti-cholinesterase activity but presented strong antidiabetic activity, in the order: 'Arbequina'≈'Picual'≈'Cuquillo'>'Hojiblanca'>'Cornicabra'. The samples showed a higher in vitro hypoglycemic effect than individual or mixed standards, possibly due to interaction between multiple identified compounds and/or a very complex multivariate interaction between other factors., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Recently, it has been reported that treatment with testosterone (T) could have favourable effects on prostate inflammation; however, the data appear inconsistent. The main evidences concern experimental studies, and there is lower information obtainable from clinical studies. This study was conducted on patients with diagnosis of male accessory gland infection (MAGI) and a concomitant hormonal condition of acquired hypergonadotropic hypogonadism and has evaluated the effects on sperm parameters of the administration of a transdermal formulation of T gel for 3 months. The treated patients showed a significantly increased percentage of spermatozoa with normal form and progressive motility (p < .05 vs baseline), a significant reduction of CD45pos leucocytes in the semen (p < .05 vs baseline) and finally a significant increase of the seminal concentrations of zinc, fructose and alpha-glucosidase (p < .05 vs baseline) identified as key parameters associated to secretory function of the male accessory glands. The results of this study suggest the use of transdermal T in hypogonadal patients with MAGI for favourable effects on sperm parameters., (© 2016 Blackwell Verlag GmbH.)

Tartary buckwheat tea (TBT) is becoming increasingly popular for its unique flavor and health benefits. However, there is little information regarding the differences in nutritional quality among different kinds of TBT. Two kinds of TBT samples, whole grain tea (WGT) and whole plant tea (WPT), were collected from markets in China. The content of their nutritional compositions, including protein, amino acids, starch, fat, fatty acids, and flavonoids, as well as aroma compounds, were determined. Antioxidant and α-glucosidase inhibitory activities were also evaluated. WPT showed higher levels of protein and fat relative to WGT, but the content of essential amino acids and fatty acids was lower. WPT contained more total flavonoids with quercetin as the predominant flavonoid (81%), while rutin was the principal flavonoid compound (86%) in WGT. GC-MS analysis indicated that there were some differences in the aroma compound profile between WGT and WPT, though 14 aroma compounds were identified in both of them with 3-ethyl-2,5-dimethyl-pyrazine as the predominant compound. In addition, WPT showed superior antioxidant and α-glucosidase inhibitory activities than WGT. However, the total flavonoid content determined in tea soup was decreased by 60% and 98% compared with that in the alcohol extracts of WGT and WPT, respectively. The antioxidant and α-glucosidase inhibitory activities of tea soup were also lowered, especially for WPT. The differences between 2 kinds of TBT might be due to the variety of raw materials and processing methods. Diversified plant materials and scientific processing technologies should be taken into account to develop a nutritional and healthy TBT product., (© 2017 Institute of Food Technologists®.)

Recently, α-glucosidase inhibitor has been widely used in clinic for diabetic therapy. In the present study, a facile and sensitive fluorescent assay based on enzyme activated inner filter effect (IFE) on nitrogen-doped carbon dots (CDs) was first developed for the detection of α-glucosidase. The N-doped CDs with green emission were prepared by a one-step hydrothermal synthesis and gave the fluorescence quantum yield of 30%, which were used as the signal output. Through α-glucosidase catalysis, 4-nitrophenol was released from 4-nitrophenyl-α-d-glucopyranoside (NGP). Interestingly, the absorption of 4-nitrophenol and the excitation of CDs were completely overlapping. Due to its great molar absorptivity, 4-nitrophenol was capable of acting as a powerful absorber to affect the fluorescent signal of CDs (i.e. IFE). By converting the absorption signals into fluorescence signals, the facile fluorescence assay strategy could be realized for α-glucosidase activity sensing, which effectively avoided the complex modification of the surface of CDs or construction of the nanoprobes. The established IFE-based sensing platform offered a low detection limit of 0.01 U/mL (S/N = 3). This proposed sensing approach has also been expanded to the inhibitor screening and showed excellent applicability. As a typical α-glucosidase inhibitor, acarbose was investigated with a low detection limit of 10 -8  M. This developed method enjoyed many merits including simplicity, lost cost, high sensitivity, good reproducibility and excellent selectivity, which also provided a new insight on the application of CDs to develop the facile and sensitive biosensor., (Copyright © 2017 Elsevier B.V. All rights reserved.)

This study aimed to investigate the effects of different concentrations of sodium bicarbonate (NaHCO 3 ) on the accumulation of flavonoids, total phenolics and d-chiro-inositol (DCI), as well as the antioxidant and α-glucosidase inhibitory activities, in tartary buckwheat sprouts. Treatment with low concentrations of NaHCO 3 (0.05, 0.1, and 0.2%) resulted in an increase in flavonoids, total phenolic compounds and DCI concentrations, and improved DPPH radical-scavenging and α-glucosidase inhibition activities compared with the control (0%). The highest levels of total flavonoids (26.69mg/g DW), individual flavonoids (rutin, isoquercitrin, quercetin, and kaempferol), total phenolic compounds (29.31mg/g DW), DCI (12.56mg/g DW), as well as antioxidant and α-glucosidase inhibition activities, were observed in tartary buckwheat sprouts treated with 0.05% NaHCO 3 for 96h. These results indicated that appropriate treatment with NaHCO 3 could improve the healthy benefits of tartary buckwheat sprouts., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Phenolics from free and hydrolyzed fractions of pomegranate juice (PJ) and seeds (PS) were evaluated. In general, total phenolic contents and scavenging of ABTS + , DPPH and hydroxyl radicals, as well as metal chelation of the soluble fraction from PS, were higher than those for PJ. Insoluble-bound phenolics from PS accounted for up to 27% of total scavenging capacity (free+esterified+insoluble-bound). Phenolic acids (13), monomeric flavonoids (8), hydrolysable tannins (12), proanthocyanidin (1) and anthocyanins (12) were tentatively characterized using HPLC-DAD-ESI-MS n . Several compounds were identified for the first time in PJ or PS. The inhibition of DNA damage (induced by hydroxyl and peroxyl radicals), copper-induced LDL-cholesterol peroxidation, as well as alpha-glucosidase and lipase activities were demonstrated, therefore supporting the potential exploitation of PJ and PS as sources of bioactive compounds., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Sweet potato (Ipomoea batatas) leaf (SPL) is an underused commercial vegetable with considerable bio-activities. By means of DPPH scavenging ability and α-glucosidase inhibitory oriented isolation, 9 and 7 compounds were isolated and identified, respectively. Among them, trans-N-(p-coumaroyl)tyramine (1), trans-N-feruloyltyramine (2), cis-N-feruloyltyramine (3), 4,5-feruloylcourmaoylquinic acid (8), caffeic acid ethyl ester (10), 7-hydroxy-5-methoxycoumarin (11), 7,3'-dimethylquercetin (13) and indole-3-carboxaldehyde (15), were firstly identified from SPL, and four of them (1, 2, 3 and 10) were firstly identified from genus Ipomoea. Phenethyl cinnamides and 3,4,5-triCQA exhibited the strongest α-glucosidase inhibition, while 3,4,5-triCQA and diCQAs were the dominant antioxidants. Structure-activity relationship revealed that higher caffeoylation of quinic acid and lower methoxylation of flavonols resulted in stronger antioxidant activity, and methylation and cis-configuration structure of phenethyl cinnamides weaken the α-glucosidase inhibition. Aforementioned results could help to explain the antioxidant activity and anti-diabetic activity of SPL, and provide theoretical basis for its further application., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Falandi is a common strawberry (Fragaria × ananassa Duch.) cultivar in southern China. Further study of the chemical constituents in Falandi fruit led to the isolation of nine norsesquiterpenoids and three triterpenoids. Falandioside D (1) and falandins A (2) and B (3) were new norsesquiterpenoids, and the others excluding tormentic acid (11) were found in strawberry for the first time. Compounds 1 and 11 exhibited potent α-glucosidase inhibitory activity with the half maximal inhibitory concentration (IC50) values of 565.0 and 27.4 μM in comparison to acarbose (619.9 μM). Compounds 3, 7 (blumenol C glucoside), and 11 showed cytotoxicity against human nasopharyngeal carcinoma cell line CNE1 with the IC50 values of 57.6, 56.4, and 36.0 μM, respectively. Among new compounds, 1 showed 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation scavenging capacity (IC50=36.2 μM). These results suggested that non-phenolic constituents were also involved in the antidiabetic, antitumour, and antioxidant effects of strawberry fruit., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Aims: This study was to determine the sources, location and identity of α-glucosidases in dormant/germinating/outgrowing spores and growing cells of Geobacillus stearothermophilus ATCC 7953, an enzymatic activity in spores used in rapid tests of steam sterilization., Methods and Results: α-Glucosidase activity in spores and cells was determined measuring methylumbelliferyl-α-d-glucoside (α-MUG) or α-MUG-6-phosphate hydrolysis fluorometrically. While α-MUG-6-phosphate was not hydrolysed by cell or spore extracts, assays with α-MUG showed that: (1) the α-glucosidase activity was inside and outside spores, and the activity outside spores was largely removed by buffer washes or heat activation, whereas α-glucosidase activity was only inside vegetative cells; (2) most α-glucosidase activity in cells and spores was soluble; (3) Western blots and enzyme inhibition using an anti-α-glucosidase antiserum identified ≥2 α-glucosidases in spores and growing cells; (4) α-glucosidase-specific activities were similar in dormant, germinated and outgrowing spore and growing cell extracts; and (5) significant α-glucosidase was synthesized during spore germination and outgrowth and cell growth, this synthesis was not repressed by glucose nor induced by α-MUG, but glucose inhibited α-MUG uptake., Conclusions: α-MUG hydrolysis by G. stearothermophilus is by α-MUG uptake and hydrolysis by ≥2 α-glucosidases associated with dormant spores and synthesized by germinating and outgrowing spores. The enzyme activity observed by sterilization assurance assays appears likely to come from heat-stable enzyme in the spore core and enzyme(s) synthesized in spore outgrowth., Significance and Impact of the Study: The results of this work provide new insight into the science behind a rapid test for steam sterilization assurance., (© 2016 The Society for Applied Microbiology.)

Diabetes mellitus is normally characterized by chronic hyperglycemia associated with disturbances in the fat, carbohydrate, and protein metabolism. There is an increasing trend of using natural products instead of synthetic agents as alternative therapy for disorders due to their fewer side effects. In this study, antidiabetic and antioxidant activities of different Melicope lunu-ankenda (ML) ethanolic extracts were evaluated using inhibition of α-glucosidase and 2,2-diphenyl-l-picrylhydrazyl (DPPH) radicals scavenging activity, respectively; whereas, proton nuclear magnetic resonance ((1) H NMR) and ultra-high performance liquid chromatography-tandem mass spectrometric (UHPLC-MS/MS) techniques were used for metabolite profiling of ML leaf extracts at different concentrations of ethanol and water. Sixty percent of ethanolic ML extract showed highest inhibitory effect against α-glucosidase enzyme (IC50 of 37 μg/mL) and DPPH scavenging activity (IC50 of 48 μg/mL). Antidiabetic effect of ML extracts was also evaluated in vivo and it was found that the high doses (400 mg/Kg BW) of ML extract exhibited high suppression in fasting blood glucose level by 62.75%. The metabolites responsible for variation among ML samples with variable ethanolic levels have been evaluated successfully using (1) H-NMR-based metabolomics. The principal component analysis (PCA) and partial least squares(PLS) analysis scores depicted clear and distinct separations into 4 clusters representing the 4 ethanolic concentrations by PC1 and PC2, with an eigenvalue of 69.9%. Various (1) H-NMR chemical shifts related to the metabolites responsible for sample difference were also ascribed. The main bioactive compounds identified attributing toward the separation included: isorhamnetin, skimmianine, scopoletin, and melicarpinone. Hence, ML may be used as promising medicinal plant for the development of new functional foods, new generation antidiabetic drugs, as a single entity phytomedicine or in combinational therapy., (© 2016 Institute of Food Technologists®)

Unextractable phenolics from plant foods and their role in health benefits have become increasingly important. Meal residues of three quinoa seeds free of fat and extractable phenolics were subjected to acid, alkaline, and enzymatic hydrolyses. The total and individual phenolic compounds released were analyzed, and 19 phenolics, predominantly phenolic acids and several flavonoids, were identified. The concentration of bound phenolics was highest in black quinoa followed by red and white, regardless of the hydrolysis method. Higher phenolic contents also showed stronger antioxidant activities and inhibition of α-glucosidase and pancreatic lipase activities. Carbohydrases, that is, pectinase, xylanase and feruloyl esterase, which effectively liberated bound phenolics are known to be secreted by colonic bacteria, suggesting potential antioxidant and anti-inflammatory effects by these compounds in the large intestine during colonic fermentation. These results can also be applied to treat foods high in bound phenolics to enhance bioaccessibility.

Three new phenolic glucosides, falandiosides A (1), B (2), and C (6) were isolated from strawberry (Fragaria×ananassa Duch.) cv. Falandi fruit, together with three flavone glucuronides (3-5), eleven lignan glycosides (12-22), and five others. Their structures were determined by spectroscopic methods. All the known phenolics were reported from strawberry for the first time. They were evaluated for antioxidant and α-glucosidase inhibitory activities. Three new and fifteen known phenolics showed potent 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation scavenging activity with IC50 values of 22.50-4.28μM in comparison to l-ascorbic acid (14.21μM). Quercetin 3-(6-methylglucuronide) (4), (+)-isolariciresinol 9'-glucoside (12), and (-)-isolariciresinol 9'-glucoside (13) were active in scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals. Moreover, compounds 12 and 13 had moderate ferric reducing antioxidant power (FRAP) values. Further, two new and seven known phenolics exhibited more potent α-glucosidase inhibitory activity with IC50 values of 537.43-25.39μM than acarbose (619.94μM)., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

A fluorescence method was established for a α-glucosidase activity assay and inhibitor screening based on β-cyclodextrin-coated quantum dots. p-Nitrophenol, the hydrolysis product of the α-glucosidase reaction, could quench the fluorescence of β-cyclodextrin-coated quantum dots via an electron transfer process, leading to fluorescence turn-off, whereas the fluorescence of the system turned on in the presence of α-glucosidase inhibitors. Taking advantage of the excellent properties of quantum dots, this method provided a very simple, rapid and sensitive screening method for α-glucosidase inhibitors. Two α-glucosidase inhibitors, 2,4,6-tribromophenol and acarbose, were used to evaluate the feasibility of this screening model, and IC50 values of 24 μM and 0.55 mM were obtained respectively, which were lower than those previously reported. The method may have potential application in screening α-glucosidase inhibitors., (Copyright © 2015 John Wiley & Sons, Ltd.)

The development of a versatile approach for the rapid and sensitive detection of relevant pathogenic bacteria and autonomous signaling of the detection events in reporter hydrogel film coatings is reported. Exploiting chitosan hydrogel films equipped with chromogenic or fluorogenic reporter moieties, the presence of the Gram-negative bacterium Pseudomonas aeruginosa and the Gram-positive bacterium Staphylococcus aureus is sensed within 1 h by detecting the characteristic enzymes α-glucosidase and elastase with limits of detection (LOD) <45 × 10(-9) M and <20 × 10(-9) M, respectively, for this observation time. The values for the LOD are two to three orders of magnitude smaller than the concentrations of the enzymes detected in the corresponding bacterial supernatants. The results show that the covalently conjugated reporter moieties are exclusively and efficiently reacted by the associated enzyme, allowing in principle for discrimination among different types of bacteria. Since high enzyme concentrations are a result of proliferating bacteria, e.g., in wounds or food, and since the selectivity of the reporting function is easily adapted to bacteria of choice, these reporter hydrogels comprise an interesting platform for the rapid detection of bacteria., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Phytohemagglutin (PHA), purified from red kidney beans (Phaseolus vulgaris) by Affi-Gel blue affinity chromatography, was subjected to ultrahigh-pressure (UHP) treatment (150, 250, 350, and 450 MPa). The purified PHA lost its hemagglutination activity after 450 MPa treatment and showed less pressure tolerance than crude PHA. However, the saccharide specificity and α-glucosidase inhibition activity of the purified PHA did not change much after UHP treatment. Electrophoresis staining by periodic acid-Schiff (PAS) manifested that the glycone structure of purified PHA remained stable even after 450 MPa pressure treatment. However, electrophoresis staining by Coomassie Blue as well as circular dichroism (CD) and differential scanning calorimetry (DSC) assay proved that the protein unit structure of purified PHA unfolded when treated at 0-250 MPa but reaggregates at 250-450 MPa. Therefore, the hemagglutination activity tends to be affected by the protein unit structure, while the stability of the glycone structure contributed to the remaining α-glucosidase inhibition activity.

The synthesis and evaluation as activity-based probes (ABPs) of three configurationally distinct, fluorescent N-alkyl cyclophellitol aziridine isosteres for profiling GH1 β-glucosidase (GBA), GH27 α-galactosidase (GLA) and GH29 α-fucosidase (FUCA) is described. In comparison with the corresponding acyl aziridine ABPs reported previously, the alkyl aziridine ABPs are synthesized easily and are more stable in mild acidic and basic media, and are thus easier to handle. The β-glucose-configured alkyl aziridine ABP proves equally effective in labeling GBA as its N-acyl counterpart, whereas the N-acyl aziridines targeting GLA and FUCA outperform their N-alkyl counterparts. Alkyl aziridines can therefore be an attractive alternative in retaining glycosidase ABP design, but in targeting a new retaining glycosidase both N-alkyl and N-acyl aziridines are best considered at the onset of a new study., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Objective: To observe the clinical efficacy of Shugan Yiyang Capsules in the treatment of asthenospermia and its action mechanisms., Methods: We randomly assigned 135 asthenospermia patients to groups A (n = 47), B (n = 45), and C (n = 43) to be treated with Shugan Yiyang Capsules, oral levocarnitine, or combination of the two. We observed sperm quality and the level of α-glucosidase in the seminal plasma before and after medication., Results: The total effectiveness rate was 70.21% in group A (markedly effective in 16 cases and effective in 17), 68.89% in group B (markedly effective in 15 cases and effective in 16), and 83.72% in group C (markedly effective in 16 cases and effective in 20), significantly higher in C than in A and B (P < 0.05). Both sperm quality and the level of α-glucosidase in the seminal plasma were improved in the three groups of patients, most obviously in group C., Conclusion: Shugan Yiyang Capsules can be used for the treatment of asthenospermia, and its effect can be enhanced in combination with oral levocarnitine.

A new assay based on ultrafiltration, liquid chromatography and mass spectrometry was developed for the rapid screening and identification of the ligands for α-glucosidase from the extract of Panax japonicus. Six saponins were identified as α-glucosidase inhibitors. Subsequently, the specific binding ligands, namely, notoginsenoside R1 , ginsenoside Rb1 , chikusetsusaponin V, chikusetsusaponin IV, chikusetsusaponin IVa, and ginsenoside Rd (the purities were 94.18, 95.43, 96.09, 93.26, 94.50, 93.86%, respectively) were separated by counter-current chromatography using two-phase solvent systems composed of tert-butyl methyl ether, acetonitrile, 0.1% aqueous formic acid (3.8:1.0:4.4, v/v/v) and the solvent system composed of methylene chloride, isopropanol, methanol, 0.1% aqueous formic acid (5.8:1.0:6.0:2.2, v/v/v). The results demonstrate that ultrafiltration, liquid chromatography and mass spectrometry combined with high-speed counter-current chromatography might provide not only a powerful tool for screening and isolating α-glucosidase inhibitors in complex samples but also a useful platform for discovering bioactive compounds for the prevention and treatment of diabetes mellitus., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Phenolic extracts from 20 Canadian lentil cultivars (Lens culinaris) were evaluated for total phenolic contents and composition, antioxidant activities (DPPH, FRAP, ORAC), and inhibitory properties against α-glucosidase and pancreatic lipase. Twenty one phenolic compounds were identified in the present study, with the majority being flavonoids, including kaempeferol glycosides, catechin/epicatechin glucosides and procyanidins. These phenolic compounds not only contributed significantly to the antioxidant activities, but they were also good inhibitors of α-glucosidase and lipase, two enzymes, respectively, associated with glucose and lipid digestion in the human intestine, thus contributing significantly to the control of blood glucose levels and obesity. More interestingly, it was the flavonols, not the flavanols, which showed the inhibitory activities against α-glucosidase and pancreatic lipase. Our result provides supporting information for developing lentil cultivars and functional foods with improved health benefits and suggests a potential role of lentil consumption in managing weight and control of blood glucose., (Copyright © 2014. Published by Elsevier Ltd.)

Background: Novel chemical molecules recovered from endangered medicinal plants have wide applications and have the potential to cure different diseases caused by microorganisms. The aim of this study was to investigate In vitro antimicrobial, α-glucosidase inhibition and antioxidant activity of different solvent extracts of Epaltes divaricata L., Methods: Antimicrobial activity of hexane, ethyl acetate and methanol extract of Epaltes divaricata was determined against bacteria and fungi using disc diffusion and microdilution method respectively. α-glucosidase inhibition, Total phenolic content (TPC), Reducing power activity, DPPH radical scavenging assay, hydroxyl radical scavenging activity, nitric oxide scavenging activity, superoxide scavenging activity and lipid peroxidation assay of plant extracts were performed according to standard protocol. Compound detection from the potential solvent extract was done through GC-MS analysis., Results: Epaltes divaricata ethyl acetate extracts (EDEa) (1.25 mg/disc) showed significant inhibition for E. lentum (23 mm), E. aerogenes (18 mm), P. fluorescence (15 mm) and A. baumanii (15 mm). Minimum inhibitory concentration (MIC) of EDEa was found to be 31.25 μg/ml, 62.5 μg/ml and 62.5 μg/ml against A. flavus, A. niger and T. rubrum respectively. EDEa showed more α-glucosidase inhibition and antioxidant activity compared to hexane and methanol. EDEa showed 50% α-glucosidase inhibition at the concentration of 525.20 ± 2.37 μg/ml. The TPC of EDEa was 412.0 ± 2.21 mg of catechol equivalents/g extract. EDEa showed great scavenging activity on 2,2-diphenyl-picrylhydrazyl (DPPH) (IC50 560 ± 2.02 μg/ml), hydroxyl (IC50 314.75 ± 2.56 μg/ml), nitric oxide (IC50 648.20 ± 2.09 μg/ml) and superoxide (IC50 361.14 ± 1.45 μg/ml) radicals, as well as high reducing power. EDEa also showed a more suppressive effect on lipid peroxidation. Using Antioxidant β-carotene linoleate method, the scavenging values of EDEa was significantly lower than BHT. GC-MS analysis of EDEa showed maximum amount of 2-butenamide, N-(4-fluorophenyl)-3-methyl trans-cinnamyl tiglate silane and trichlorocyclohexyl silane (36.86%)., Conclusion: The results obtained in this study clearly indicate that EDEa can be used as a natural antimicrobial, α-glucosidase inhibition and antioxidant agent.

Type 2 diabetes is a metabolic disorder affecting millions of people worldwide, and new drug leads or functional foods containing selective α-glucosidase inhibitors are needed. Crude extract of 24 plants were assessed for α-glucosidase inhibitory activity. Methanol extracts of Cinnamomum zeylanicum bark, Rheum rhabarbarum peel, and Rheum palmatum root and ethyl acetate extracts of C. zeylanicum bark, Allium ascalonicum peel, and R. palmatum root showed IC50 values below 20 μg/mL. Subsequently, high-resolution α-glucosidase profiling was used in combination with high-performance liquid chromatography-high-resolution mass spectrometry-solid-phase extraction-nuclear magnetic resonance spectroscopy for identification of metabolites responsible for the α-glucosidase inhibitory activity. Quercetin (1) and its dimer (2), trimer (3), and tetramer (4) were identified as main α-glucosidase inhibitors in A. ascalonicum peel, whereas (E)-piceatannol 3'-O-β-D-glucopyranoside (5), (E)-rhapontigenin 3'-O-β-D-glucopyranoside (6), (E)-piceatannol (8), and emodin (12) were identified as main α-glucosidase inhibitors in R. palmatum root.

Objective: To investigate the differences in semen quality between samples collected by masturbation in the clinic and at home., Methods: Based on the WHO guidelines, we analyzed the ejaculates collected by masturbation in the clinic and at home from 342 men under infertility assessment and measured the contents of such biochemical markers in the seminal plasma as neutral α-glucosidase, zinc, and fructose. According to the location of semen collection, we divided the samples into two groups, clinic-collected and home-collected, and analyzed the differences in the semen parameters between the two groups with the SPSS 16.0 software., Results: Compared with the clinic-collected semen, the home-collected samples had significantly higher mean values in semen volume (4.0 vs 4.9%), sperm concentration (41 vs 64 x 10(6)/ml), total sperm count (175 vs 270 x 10(6) per ejaculate), progressive sperm motility (40 vs 52%), total count of progressively motile sperm (82 vs 135 x 10(6) per ejaculate) (all P <0.05). No significant differences were found between the two groups in normal sperm morphology (4.0 vs 5.0%) and the contents of neutral α-glucosidase (26 vs 24 mU per ejaculate), zinc (8.0 vs 8.0 μmol per ejaculate), and fructose (62 vs 60 μmol per ejaculate) (all P >0.05). Abnormal sperm concentration (<20 x 10(6)/ml) was observed in significantly fewer of the home-collected samples than the clinic-collected ones (18% [62/342] vs 30% [103/342], P<0.05), and so was abnormal progressive sperm motility (<32%) (64% [219/342] vs 75% [256/342], P<0.05)., Conclusion: Our findings show that semen samples collected by masturbation at home has a higher quality than those collected in the clinic. So the location of semen collection should be taken into consideration in infertility investigation.

The assessment of freshness of different sizes of blue fish (Engraulis encrasicolus 12 cm, Sardina pilchardus 15 cm, Trachurus trachurus 40 cm, Scomber japonicus colias 60 cm) was carried out using non-conventional enzymatic methods. The activities of the three lysosomal enzymes (α-glucosidase (AG), β-galactosidase (B-GAL) and β-N-acetylglucosamidase (B-NA)) in extracts of blue fish muscle were measured over a period of 21 days of storage. A significant increase (p<0.05) of AG activity was observed in all species, with a large increase seen after only one day of storage. B-NA activity increased slightly in sardines, horse mackerels and chub mackerel during frozen/thawed storage. Finally, the increase of B-GAL activity was significant (p<0.05) only in the samples of larger blue fish as horse mackerel and chub mackerel. All of these enzyme activities may be helpful predictive markers to limit fraud in these species., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Oestrogen biosynthesis in ejaculated spermatozoa is an autonomous process, which may influence sperm functions. The purpose of this study was to evaluate the relationship between the expression of aromatase, sperm quality and seminal neutral α-glucosidase marker in semen of Tunisian infertile men: asthenozoospermia (A; n = 16), teratozoospermia (T; n = 12) and asthenoteratozoospermia (AT; n = 11) in comparison with 18 normozoospermic ones. Aromatase mRNA levels estimated by real-time PCR were reduced in groups T (52%) and AT (67%) compared to controls and inversely correlated with the percentage of normal forms. A higher coefficient of correlation was noted in presence of microcephaly or acrosome malformations (r = -0.64). The asthenozoospermic group was divided into two subgroups according to the relative amount of aromatase. The subgroup (A2) with higher aromatase transcript level was associated with an increased seminal pH, a decreased sperm viability, low sperm percentage motility and low neutral α-glucosidase semen levels. Our data highlight the involvement of aromatase in motility and morphology of spermatozoa. Thus, this enzyme could bring new insights about quality and fertilising capacity of human spermatozoa., (© 2012 Blackwell Verlag GmbH.)