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Sample records for alpha-glucosidases

  1. Macroalage as a source of alpha-glucosidase inhibitors

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Alpha-glucosidase inhibitors were screened from organic solvent extracts of macroalgae by a spectrophotometrical method with p-nitrophenyl-D-glucopyranosidase as the substrate. The result indicates that organic crude extracts from some macroalgae such as Rhodomela confervoides (Huds.) Silva, Gracilaria textorii (Suringar) DeToni, Plocamium telfairiae Harv., Dictyopteris divaricata (Okam.) Okam, Ulval pertusa and Enteromorpha intestinalis (L.) Link et al. show strong inhibitory activity of alpha-glucosidase at concentration of 79.6 μg/ml.

  2. The Role of alpha-Glucosidase in Germinating Barley Grains

    DEFF Research Database (Denmark)

    Stanley, Duncan; Rejzek, Martin; Næsted, Henrik;

    2011-01-01

    in the endosperm 10 d after imbibition, implying inhibition of maltase activity. Three of the four inhibitors also reduced starch degradation and seedling growth, but the fourth did not affect these parameters. Inhibition of starch degradation was apparently not due to inhibition of amylases. Inhibition......The importance of alpha-glucosidase in the endosperm starch metabolism of barley (Hordeum vulgare) seedlings is poorly understood. The enzyme converts maltose to glucose (Glc), but in vitro studies indicate that it can also attack starch granules. To discover its role in vivo, we took complementary...... silencing cassette for HvAgl97, alpha-glucosidase activity was reduced by up to 50%. There was a large decrease in the Glc-to-maltose ratio in these lines but no effect on starch degradation or seedling growth. Our results suggest that the alpha-glucosidase HvAGL97 is the major endosperm enzyme catalyzing...

  3. A novel alpha-glucosidase from the moss Scopelophila cataractae.

    Science.gov (United States)

    Yamasaki, Yoshiki; Nakashima, Susumu; Konno, Haruyoshi

    2007-01-01

    Scopelophila cataractae is a rare moss that grows on copper-containing soils. S. cataractae protonema was grown on basal MS medium containing copper. A starch-degrading activity was detected in homogenates of the protonema, after successive extraction with phosphate buffer and buffer containing 3 M LiCl. Buffer-soluble extract (BS) and LiCl-soluble extract (LS) readily hydrolyzed amylopectin to liberate only glucose, which shows that alpha-glucosidase (EC 3.2.1.20) in BS and LS hydrolyzed amylopectin. The K(m) value of BS for maltose was 0.427. The K(m) value of BS for malto-oligosaccharide decreased with an increase in the molecular mass of the substrate. The value for maltohexaose was 0.106, which is about four-fold lower than that for maltose. BS was divided into two fractions of alpha-glucosidase (BS-1 and BS-2) by isoelectric focusing. The isoelectric points of these two enzymes were determined to be 4.36 (BS-1) and 5.25 (BS-2) by analytical gel electrofocusing. The two enzymes readily hydrolyzed malto-oligosaccharides. The two enzymes also hydrolyzed amylose, amylopectin and soluble starch at a rate similar to that with maltose. The two enzymes readily hydrolyzed panose to liberate glucose and maltose (1 : 1), and the K(m) value of BS for panose was similar to that for maltotriose, whereas the enzymes hydrolyzed isomaltose only weakly. With regard to substrate specificity, the two enzymes in BS are novel alpha-glucosidases. The two enzymes also hydrolyzed beta-limit dextrin, which has many alpha-1,6-glucosidic linkages near the non-reducing ends, more strongly than maltose, which shows that they do not need a debranching enzyme for starch digestion. The starch-degrading activity of BS was not inhibited by p-chloromercuribenzoic acid or alpha-amylase inhibitor. When amylopectin was treated with BS and LS in phosphate buffer, pH 6.0, glucose, but not glucose-1-phosphate, was detected, showing that the extracts did not contain phosphorylase but did contain an

  4. A specific acid [alpha]-glucosidase in lamellar bodies of the human lung

    OpenAIRE

    Vries, A.C.J. de; Schram, A.W.; Tager, J.M.; Batenburg, J.J.

    2006-01-01

    In the present investigation, we have demonstrated that three lysosomal-type hydrolases, alpha-glucosidase, alpha-mannosidase and a phosphatase, are present in lamellar bodies isolated from adult human lung. The hydrolase activities that were studied, all showed an acidic pH optimum, which is characteristic for lysosomal enzymes. The properties of acid alpha-glucosidase in the lamellar body fraction and that in the lysosome-enriched fraction were compared. Using specific antibodies against ly...

  5. The role of alpha-glucosidase in germinating barley grains.

    Science.gov (United States)

    Stanley, Duncan; Rejzek, Martin; Naested, Henrik; Smedley, Mark; Otero, Sofía; Fahy, Brendan; Thorpe, Frazer; Nash, Robert J; Harwood, Wendy; Svensson, Birte; Denyer, Kay; Field, Robert A; Smith, Alison M

    2011-02-01

    The importance of α-glucosidase in the endosperm starch metabolism of barley (Hordeum vulgare) seedlings is poorly understood. The enzyme converts maltose to glucose (Glc), but in vitro studies indicate that it can also attack starch granules. To discover its role in vivo, we took complementary chemical-genetic and reverse-genetic approaches. We identified iminosugar inhibitors of a recombinant form of an α-glucosidase previously discovered in barley endosperm (ALPHA-GLUCOSIDASE97 [HvAGL97]), and applied four of them to germinating grains. All four decreased the Glc-to-maltose ratio in the endosperm 10 d after imbibition, implying inhibition of maltase activity. Three of the four inhibitors also reduced starch degradation and seedling growth, but the fourth did not affect these parameters. Inhibition of starch degradation was apparently not due to inhibition of amylases. Inhibition of seedling growth was primarily a direct effect of the inhibitors on roots and coleoptiles rather than an indirect effect of the inhibition of endosperm metabolism. It may reflect inhibition of glycoprotein-processing glucosidases in these organs. In transgenic seedlings carrying an RNA interference silencing cassette for HvAgl97, α-glucosidase activity was reduced by up to 50%. There was a large decrease in the Glc-to-maltose ratio in these lines but no effect on starch degradation or seedling growth. Our results suggest that the α-glucosidase HvAGL97 is the major endosperm enzyme catalyzing the conversion of maltose to Glc but is not required for starch degradation. However, the effects of three glucosidase inhibitors on starch degradation in the endosperm indicate the existence of unidentified glucosidase(s) required for this process.

  6. Cuminaldehyde: Aldose Reductase and alpha-Glucosidase Inhibitor Derived from Cuminum cyminum L. Seeds.

    Science.gov (United States)

    Lee, Hoi-Seon

    2005-04-06

    The inhibitory activity of Cuminum cyminum seed-isolated component was evaluated against lens aldose reductase and alpha-glucosidase isolated from Sprague-Dawley male rats and compared to that of 11 commercially available components derived from C. cyminum seed oil, as well as quercitrin as an aldose reductase inhibitor and acarbose as an alpha-glucosidase inhibitor. The biologically active constituent of C. cyminum seed oil was characterized as cuminaldehyde by various spectral analyses. The IC(50) value of cuminaldehyde is 0.00085 mg/mL against aldose reductase and 0.5 mg/mL against alpha-glucosidase, respectively. Cuminaldehyde was about 1.8 and 1.6 times less in inhibitory activity than acarbose and quercitin, respectively. Nonetheless, cuminaldehyde may be useful as a lead compound and a new agent for antidiabetic therapeutics.

  7. Unexpected high digestion rate of cooked starch by the Ct-Maltase-Glucoamylase small intestine mucosal alpha-glucosidase subunit

    Science.gov (United States)

    For starch digestion to glucose, two luminal alpha-amylases and four gut mucosal alpha-glucosidase subunits are employed. The aim of this research was to investigate, for the first time, direct digestion capability of individual mucosal alpha-glucosidases on cooked (gelatinized) starch. Gelatinized ...

  8. A specific acid [alpha]-glucosidase in lamellar bodies of the human lung

    NARCIS (Netherlands)

    Vries, A.C.J. de; Schram, A.W.; Tager, J.M.; Batenburg, J.J.

    2006-01-01

    In the present investigation, we have demonstrated that three lysosomal-type hydrolases, alpha-glucosidase, alpha-mannosidase and a phosphatase, are present in lamellar bodies isolated from adult human lung. The hydrolase activities that were studied, all showed an acidic pH optimum, which is charac

  9. Effective Control of Postprandial Glucose Level through Inhibition of Intestinal Alpha Glucosidase by Cymbopogon martinii (Roxb.

    Directory of Open Access Journals (Sweden)

    Varsha Ghadyale

    2012-01-01

    Full Text Available Inhibition of intestinal alpha glucosidase plays a major role in preventing rise in postprandial glucose level in diabetics. Cymbopogon martinii (CM (family Poaceae is used in traditional Indian medicine in treatment of diabetes mellitus. The alpha glucosidase inhibitory action of the plant is studied. The active component was separated using hot water extraction of the whole plant powder, differential solvent extraction, and silica gel column chromatography. The 30 : 70 toluene : ethyl acetate fraction showed optimum activity. The silica gel chromatography fraction demonstrated 98, 98, and 68% inhibition for starch, maltose, and sucrose, respectively, at 5 mg/kg body weight of rats. Intestinal absorption studies using noneverted intestinal sacs, as well as in vivo studies in streptozotocin-induced diabetic rats using oral glucose tolerance with maltose and sucrose load, revealed better inhibition of alpha glucosidase as compared to acarbose. Kinetic studies using Lineweaver Burk plot showed mixed to noncompetitive type of inhibition by CM. In vivo studies with maltose load of 2 mg and 3 mg/gm body weight showed a noncompetitive pattern of inhibition at 5 mg/kg body weight of CM as against 60 mg/kg body weight of acarbose. Thus CM is more effective alpha glucosidase inhibitor and at lower concentration than acarbose.

  10. Purification and partial characterization of three forms of alpha-glucosidase from the fruit fly Drosophila melanogaster.

    Science.gov (United States)

    Tanimura, T; Kitamura, K; Fukuda, T; Kikuchi, T

    1979-01-01

    Three forms of alpha-glucosidase, I, II, and III, have been purified from the whole body extract of adult flies of Drosophila melanogaster in yields of 2.1, 5.3, and 6.7%, respectively. The purification procedures involved ammonium sulfate fractionation, Con A-Sepharose 4B affinity chromatography, DEAE-Sepharose CL-6B ion exchange chromatography, Sephacryl S-200 gel filtration, and preparative gel electrophoresis. Each purified enzyme showed a single band on polyacrylamide gel on both protein and enzyme activity staining. The molecular weights of alpha-glucosidases I, II, and III were estimated to be 200,000, 56,000, and 76,000, respectively, by gel filtration. SDS gels indicated that alpha-glucosidases II and III were each composed of a single polypeptide chain, whereas alpha-glucosidase I was composed of two identical subunits. Both alpha-glucosidases II and III hydrolyzed sucrose and p-nitrophenyl-alpha-D-glucoside (PNPG), but alpha-glucosidase I hydrolyzed PNPG to a much lesser extent than sucrose. For sucrose the pH optima of alpha-glucosidases I, II, and III were pH 6.0, 5.0, and 6.0 and the Km values were 13.1, 8.9, and 10 mM, respectively. For PNPG the pH optima of alpha-glucosidases II and III were pH 5.5 and 6.5 and the Km values were 0.77 and 0.21 mM, respectively.

  11. Effects of alpha-glucosidase inhibitors on mouth to caecum transit time in humans.

    OpenAIRE

    Ladas, S D; Frydas, A; Papadopoulos, A.; S. A. Raptis

    1992-01-01

    The alpha-glucosidase inhibitors acarbose and miglitol have been successfully used to control postprandial hyperglycaemia in diabetics. They probably work by slowing carbohydrate digestion and absorption, but their effect on mouth to caecum transit time has not been studied. The effect acarbose (100 mg), miglitol (100 mg), and placebo on mouth to caecum transit time (380 kcal breakfast with 20 g of lactulose) was investigated in 18 normal volunteers using breath hydrogen analysis. Both miglit...

  12. Oligomeric procyanidins of French maritime pine bark extract (Pycnogenol) effectively inhibit alpha-glucosidase.

    Science.gov (United States)

    Schäfer, Angelika; Högger, Petra

    2007-07-01

    The standardized maritime pine bark extract (Pycnogenol) was reported to exert clinical anti-diabetic effects after peroral intake. However, an increased insulin secretion was not observed after administration of the extract to patients. Our aim was to elucidate whether the described clinical effects of Pycnogenol are related to inhibition of alpha-glucosidase. Therefore, we analyzed the inhibitory activity of Pycnogenol, green tea extract and acarbose towards alpha-glucosidase. Furthermore, we explored different fractions of Pycnogenol containing compounds of diverse molecular masses from polyphenolic monomers, dimers and higher oligomers to uncover which components exhibited the most pronounced inhibitory activity. We found that Pycnogenol exhibited the most potent inhibition (IC(50) about 5 microg/mL) on alpha-glucosidase compared to green tea extract (IC(50) about 20 microg/mL) and acarbose (IC(50) about 1mg/mL). The inhibitory action of Pycnogenol was stronger in extract fractions containing higher procyanidin oligomers. The results obtained assign a novel, local effect to oligomeric procyanidins and contribute to the explanation of glucose-lowering effects of Pycnogenol observed in clinical trials with diabetic patients.

  13. Antidiabetic property of Symplocos cochinchinensis is mediated by inhibition of alpha glucosidase and enhanced insulin sensitivity.

    Directory of Open Access Journals (Sweden)

    Kalathookunnel Antony Antu

    Full Text Available The study is designed to find out the biochemical basis of antidiabetic property of Symplocos cochinchinensis (SC, the main ingredient of 'Nisakathakadi' an Ayurvedic decoction for diabetes. Since diabetes is a multifactorial disease, ethanolic extract of the bark (SCE and its fractions (hexane, dichloromethane, ethyl acetate and 90% ethanol were evaluated by in vitro methods against multiple targets relevant to diabetes such as the alpha glucosidase inhibition, glucose uptake, adipogenic potential, oxidative stress, pancreatic beta cell proliferation, inhibition of protein glycation, protein tyrosine phosphatase-1B (PTP-1B and dipeptidyl peptidase-IV (DPP-IV. Among the extracts, SCE exhibited comparatively better activity like alpha glucosidase inhibition (IC50 value-82.07 ± 2.10 µg/mL, insulin dependent glucose uptake (3 fold increase in L6 myotubes, pancreatic beta cell regeneration in RIN-m5F (3.5 fold increase and reduced triglyceride accumulation (22% decrease in 3T3L1 cells, protection from hyperglycemia induced generation of reactive oxygen species in HepG2 cells (59.57% decrease with moderate antiglycation and PTP-1B inhibition. Chemical characterization by HPLC revealed the superiority of SCE over other extracts due to presence and quantity of bioactives (beta-sitosterol, phloretin 2'glucoside, oleanolic acid in addition to minerals like magnesium, calcium, potassium, sodium, zinc and manganese. So SCE has been subjected to oral sucrose tolerance test to evaluate its antihyperglycemic property in mild diabetic and diabetic animal models. SCE showed significant antihyperglycemic activity in in vivo diabetic models. We conclude that SC mediates the antidiabetic activity mainly via alpha glucosidase inhibition, improved insulin sensitivity, with moderate antiglycation and antioxidant activity.

  14. Effects of alpha-glucosidase inhibitors on mouth to caecum transit time in humans.

    Science.gov (United States)

    Ladas, S D; Frydas, A; Papadopoulos, A; Raptis, S A

    1992-09-01

    The alpha-glucosidase inhibitors acarbose and miglitol have been successfully used to control postprandial hyperglycaemia in diabetics. They probably work by slowing carbohydrate digestion and absorption, but their effect on mouth to caecum transit time has not been studied. The effect acarbose (100 mg), miglitol (100 mg), and placebo on mouth to caecum transit time (380 kcal breakfast with 20 g of lactulose) was investigated in 18 normal volunteers using breath hydrogen analysis. Both miglitol and acarbose significantly increased breath hydrogen excretion (F2,34 = 6.31, p = 0.005) and shortened the mouth to caecum transit time (F2,34 = 3.49, p = 0.04) after breakfast compared with placebo. There was a significant negative correlation between breath hydrogen excretion and mouth to caecum transit time suggesting that with shorter transit times significantly more carbohydrates were spilled into the colon. These results indicate that alpha-glucosidase inhibitors accelerate mouth to caecum transit time by inducing carbohydrate malabsorption.

  15. Impaired performance of skeletal muscle in alpha-glucosidase knockout mice.

    Science.gov (United States)

    Hesselink, Reinout P; Gorselink, Marchel; Schaart, Gert; Wagenmakers, Anton J M; Kamphoven, Joep; Reuser, Arnold J J; Van Der Vusse, Ger J; Drost, Maarten R

    2002-06-01

    Glycogen storage disease type II (GSD II) is an inherited progressive muscle disease in which lack of functional acid alpha-glucosidase (AGLU) results in lysosomal accumulation of glycogen. We report on the impact of a null mutation of the acid alpha-glucosidase gene (AGLU(-/-)) in mice on the force production capabilities, contractile mass, oxidative capacity, energy status, morphology, and desmin content of skeletal muscle. Muscle function was assessed in halothane-anesthetized animals, using a recently designed murine isometric dynamometer. Maximal torque production during single tetanic contraction was 50% lower in the knockout mice than in wild type. Loss of developed torque was found to be disproportionate to the 20% loss in muscle mass. During a series of supramaximal contraction, fatigue, expressed as percentile decline of developed torque, did not differ between AGLU(-/-) mice and age-matched controls. Muscle oxidative capacity, energy status, and protein content (normalized to either dry or wet weight) were not changed in knockout mice compared to control. Alterations in muscle cell morphology were clearly visible. Desmin content was increased, whereas alpha-actinin was not. As the decline in muscle mass is insufficient to explain the degree in decline of mechanical performance, we hypothesize that the large clusters of noncontractile material present in the cytoplasm hamper longitudinal force transmission, and hence muscle contractile function. The increase in muscular desmin content is most likely reflecting adaptations to altered intracellular force transmission.

  16. An alpha-glucosidase inhibitor from an endophytic Cladosporium sp. with potential as a biocontrol agent.

    Science.gov (United States)

    Singh, Bahaderjeet; Kaur, Tamanreet; Kaur, Sanehdeep; Manhas, Rajesh K; Kaur, Amarjeet

    2015-02-01

    This study highlights the importance of alpha-glucosidase inhibitors as mechanisms for endophyte-mediated resistance to insect pests. One of the major benefits which endophytes confer on plants is providing resistance against insect pests. This built-in defense mechanism of the plant can be used for exploring ecofriendly strategies for pest control. In the present study, 34 endophytic fungi were isolated from Tinospora cordifolia and screened for their ability to produce alpha-glucosidase inhibitors. Maximum inhibitory activity was observed in an isolate from T. cordifolia (TN-9S), identified to be Cladosporium sp. The inhibitor was purified using chromatographic techniques. The insecticidal activity of the purified inhibitor was evaluated against Spodoptera litura. The inhibitor induced a significant mortality in the larvae of S. litura and adversely affected its survival and development. It also inhibited the activity of α-glycosidases in vivo in the gut of the larvae. The purified inhibitor was determined to be a phenolic compound with amine groups, demonstrating a noncompetitive type of inhibition in vitro. The production of the inhibitor was optimized. Response surface methodology (RSM) analysis revealed a significant interaction between dextrose and malt extract, with first-order effect of pH.

  17. Human acid alpha-glucosidase from rabbit milk has therapeutic effect in mice with glycogen storage disease type II

    NARCIS (Netherlands)

    A.G.A. Bijvoet (Agnes); A.J.J. Reuser (Arnold); H. van Hirtum (Hans); M.A. Kroos (Marian); E.H. van de Kamp; O. Schoneveld; P. Visser (Pim); J.P. Brakenhoff (Just); M. Weggeman; E.J.J.M. van Corven (Emiel); A.T. van der Ploeg (Ans)

    1999-01-01

    textabstractPompe's disease or glycogen storage disease type II (GSDII) belongs to the family of inherited lysosomal storage diseases. The underlying deficiency of acid alpha-glucosidase leads in different degrees of severity to glycogen storage in heart, skeletal and s

  18. Screening of endophytic fungi having ability for antioxidative and alpha-glucosidase inhibitor activities isolated from Taxus sumatrana.

    Science.gov (United States)

    Artanti, N; Tachibana, S; Kardono, L B S; Sukiman, H

    2011-11-15

    Endophytic microbes are considered as an important source of natural products. They show antibiotic, anticancer, antioxidative and antidiabetic activities. Therefore, there are many reports on the isolation and bioactivity screening of endophytic fungi from various plants including Taxus species. Taxus sumatrana (Miq.) de Laub is found in Indonesia. The objective of this study is to conduct an in vitro screening of 14 endophytic fungi isolated from Taxus sumatrana having antioxidative and alpha-glucosidase inhibitor activities. Each endophytic fungus was cultured for 7 days and the fungal mycelium and medium were extracted with methanol and ethyl acetate, respectively, to produce each extract. The antioxidative activity of each extract was tested by DPPH free radical scavenging activity and beta-carotene bleaching assays, whereas antidiabetic activity was tested based on alpha-glucosidase inhibitor activity. The screening results showed that fungal mycelia of TSC 13 had the best alpha-glucosidase inhibitor activity and TSC 24 had the best antioxidative activity. Isolation of bioactive compounds from TSC 13 and TSC 24 is being conducted. This is the first report that endophytic fungi isolated from T. sumatrana exhibited anti alpha-glucosidase inhibitory and anti oxidative activities.

  19. Modulation of starch digestion for slow glucose release through "toggling" of activities of mucosal "alpha"-glucosidases

    Science.gov (United States)

    Starch digestion involves the breakdown by alpha-amylase to small linear and branched malto-oligosaccharides, which are in turn hydrolyzed to glucose by the mucosal alpha-glucosidases, maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI). MGAM and SI are anchored to the small intestinal brush-bor...

  20. alpha-Glucosidase inhibitory pentacyclic triterpenes from the stem bark of Fagara tessmannii (Rutaceae).

    Science.gov (United States)

    Mbaze, Luc Meva'a; Poumale, Herve Martial P; Wansi, Jean Duplex; Lado, Jean Alexandre; Khan, Shamsun Nahar; Iqbal, Muhammad Choudhary; Ngadjui, Bonaventure Tchaleu; Laatsch, Hartmut

    2007-03-01

    In addition to fatty acids, a mixture of sterols (beta-sitosterol, stigmasterol, campesterol and stigmastanol), lupeol, arctigenin methylether, sesamin, vanillic acid (1), 2,6-dimethoxy-1,4-benzoquinone (2), betulinic acid and two pentacyclic triterpene acetates were isolated from Fagara tessmannii Engl. They were identified as 3beta-acetoxy-16beta-hydroxybetulinic acid (3a) and 3beta,16beta-diacetoxybetulinic acid (3b), and their structures were established using 1 and 2D NMR spectra and by comparison with published data. Two derivatives of the compounds were prepared. Some isolated compounds were evaluated for their antifungal and antibacterial activities. Compounds 1 and 3a showed significant inhibition of alpha-glucosidase.

  1. Bio-assay guided isolation of alpha-glucosidase inhibitory constituents from Eclipta alba.

    Science.gov (United States)

    Kumar, Deepak; Gaonkar, Raghuvir H; Ghosh, Rina; Pal, Bikas C

    2012-08-01

    Eclipta alba (L.) Hassk is used traditionally in diabetes mellitus in India and the plant extract is reported to possess anti-diabetic activity. A bioactivity-guided isolation approach based on alpha-glucosidase inhibition was used to identify the constituents contributing towards the inhibition of the enzyme and probably contributing towards its anti-diabetic activity. Four echinocystic acid glycosides were thus isolated, of which eclalbasaponin VI, isolated from the n-butanol fraction, was found to be the most potent (IC50 54.2 +/- 1.3 microM). The compound is an uncompetitive type of inhibitor with Ki 26.1 microM. A quantitative estimation of the constituents was established using RP-HPLC.

  2. AGI, a previously unreported D. melanogaster {alpha}-glucosidase: Partial purification, characterization, and cytogenetic mapping

    Energy Technology Data Exchange (ETDEWEB)

    Parker, G.F.; Roberts, D.B. [Univ. of Oxford (United Kingdom)

    1996-04-01

    Inbred Drosophila melanogaster stocks were surveyed for {alpha}-glucosidases with nondenaturing gel electrophoresis using a fluorogenic substrate to stain the gels. The glucosidase most active under these conditions is polymorphic. We established that the polymorphism is genetic in origin and that the glucosidase was not likely to be a previously characterized enzyme. The gene encoding the enzyme was mapped cytogenetically to 33 A1-2- 33A8-B1, confirming that this is an enzyme not yet reported in D. melanogaster. The enzyme was partially purified by elution from nondenaturing gels, which enable us to establish that it has optimal activity at pH 6 and interacts most strongly with {alpha}- 1 -4 glucosides. A developmental and tissue survey suggested that this enzyme could have a purely digestive role or be involved in carbohydrate metabolism inside the organism. We propose that this enzyme is involved in either starch digestion or glycogen metabolism. 37 refs., 6 figs., 1 tab.

  3. AGI, a previously unreported D. melanogaster alpha-glucosidase: partial purification, characterization, and cytogenetic mapping.

    Science.gov (United States)

    Parker, G F; Roberts, D B

    1996-04-01

    Inbred Drosophila melanogaster stocks were surveyed for alpha-glucosidases with nondenaturing gel electrophoresis using a fluorogenic substrate to stain the gels. The glucosidase most active under these conditions is polymorphic. We established that the polymorphism is genetic in origin and that the glucosidase was not likely to be a previously characterized enzyme. The gene encoding the enzyme was mapped cytogenetically to 33 A1-2- 33A8-B1, confirming that this is an enzyme not yet reported in D. melanogaster. The enzyme was partially purified by elution from nondenaturing gels, which enabled us to establish that it has optimal activity at pH 6 and interacts most strongly with alpha-1-4 glucosides. A developmental and tissue survey suggested that this enzyme could have a purely digestive role or be involved in carbohydrate metabolism inside the organism. We propose that this enzyme is involved in either starch digestion or glycogen metabolism.

  4. [Chemical constituents from Psoralea corylifolia and their antioxidant alpha-glucosidase inhibitory and antimicrobial activities].

    Science.gov (United States)

    Wang, Tian-Xiao; Yin, Zhen-Hua; Zhang, Wei; Peng, Tao; Kang, Wen-Yi

    2013-07-01

    Twelve compounds were isolated from Psoralea corylifolia and their structures were identified as isopsoralen (1), psoralen (2), 8-methoxypsoralen (3), psoralidin (4), corylin (5), bavachin (6), daidzein (7), corylifolinin (8), bavachinin (9), neobavaisoflavone (10), daidzin (11) and astragalin (12). The results showed that psoralidin had the activity of scavenging DPPH free radicals activity (IC50 43.85 mg x L(-1)). Psoralidin (IC50 1.32 mg x L(-1))c, oryfolin (IC50 4.97 mg x L(-1)), daidzin (IC50 10.47 mg x S(-1)), daidzein (IC50 34.22 mg) x L(-1)) and astragalin (IC50 31.27 mg x L(-1)) had the activity of scavenging ABTS free radical. Psoralidin (IC50 40.74 mg x L(-1)), coryfolin (IC50 45.73 mg x L(-1)) and daidzein (IC50 49.44 mg x L(-1)) had alpha-glucosidase inhibitory activity. Corylifolinin and neobavaisoflavone had significantly effect of inhibiting SA, MRSA and ESBLs-SA (MIC 0. 781 3, 1.562, 5, 0.781 25 microg x disc(-1) and 6.25, 6.25, 6.25 microg x disc(-1).

  5. Alpha-glucosidase inhibitory and antiplasmodial properties of terpenoids from the leaves of Buddleja saligna Willd.

    Science.gov (United States)

    Chukwujekwu, Jude C; Rengasamy, Kannan R R; de Kock, Carmen A; Smith, Peter J; Slavětínská, Lenka Poštová; van Staden, Johannes

    2016-01-01

    In our continuing search for biologically active natural product(s) of plant origin, Buddleja saligna, a South African medicinal plant, was screened in line with its traditional use for antidiabetic (yeast alpha glucosidase inhibitory) and antiplasmodial (against a chloroquine sensitive strain of Plasmodium falciparum (NF54)) activities. The hexane fraction showed the most promising activity with regards to its antidiabetic (IC(50) = 260 ± 0.112 µg/ml) and antiplasmodial (IC(50) = 8.5 ± 1.6 µg/ml) activities. Using activity guided fractionation three known terpenoids (betulonic acid, betulone and spinasterol) were isolated from this species for the first time. The compounds displayed varying levels of biological activities (antidiabetic: 27.31 µg/ml ≥ IC(50) ≥ 5.6 µg/ml; antiplasmodial: 14 µg/ml ≥ IC(50) ≥ 2 µg/ml) with very minimal toxicity.

  6. Cloning the mouse homologue of the human lysosomal acid {alpha}-glucosidase gene

    Energy Technology Data Exchange (ETDEWEB)

    Ding, J.H.; Yang, B.Z.; Liu, H.M. [Duke Univ. Medical Center, Durham, NC (United States)] [and others

    1994-09-01

    Pompe disease (GSD II) is an autosomal recessive disorder caused by a deficiency of lysosomal acid {alpha}-glucosidase (GAA). In an attempt to create a mouse model for Pompe disease, we isolated and characterized the gene encoding the mouse homologue of the human GAA. Twenty clones that extend from exon 2 to the poly(A) tail were isolated from a mouse liver cDNA library, but the remainder of the mRNA proved difficult to obtain by conventional cDNA library screening. Sequences spanning exons 1-2 were cloned by RACE from mouse liver RNA. The full-length liver GAA cDNA contains 3365 nucleotides with a coding region of 2859 nucleotides and a 394 base pair 3{prime}-nontranslated region. The deduced amino acid sequence of the mouse GAA shows 84% identity to the human GAA. Southern blot analysis demonstrated that the mouse GAA was encoded by a single copy gene. Then six bacteriophages containing DNA from the GAA gene were isolated by screening 10{sup 6} phage plaques of a mouse 129 genomic library using a mouse GAA cDNA as a probe. From one of these bacteriophages, an 11-kilobase EcoRI fragment containing exons 3 to 15 was subcloned and sequenced. Work is in progress using this genomic clone to disrupt the GAA gene in murine embryonic stem cells in order to create GSD II mice.

  7. Inhibitory Properties of Aqueous Ethanol Extracts of Propolis on Alpha-Glucosidase

    Directory of Open Access Journals (Sweden)

    Hongcheng Zhang

    2015-01-01

    Full Text Available The objective of the present study was to evaluate the inhibitory properties of various extracts of propolis on alpha-glucosidase from baker’s yeast and mammalian intestine. Inhibitory activities of aqueous ethanol extracts of propolis were determined by using 4-nitrophenyl-D-glucopyranoside, sucrose and maltose as substrates, and acarbose as a positive reference. All extracts were significantly effective in inhibiting α-glucosidase from baker’s yeast and rat intestinal sucrase in comparison with acarbose (P<0.05. The 75% ethanol extracts of propolis (75% EEP showed the highest inhibitory effect on α-glucosidase and sucrase and were a noncompetitive inhibition mode. 50% EEP, 95%, EEP and 100% EEP exhibited a mixed inhibition mode, while water extracts of propolis (WEP and 25% EEP demonstrated a competitive inhibition mode. Furthermore, WEP presented the highest inhibitory activity against maltase. These results suggest that aqueous ethanol extracts of propolis may be used as nutraceuticals for the regulation of postprandial hyperglycemia.

  8. Evaluation of anti-diabetic and alpha glucosidase inhibitory action of anthraquinones from Rheum emodi.

    Science.gov (United States)

    Arvindekar, Aditya; More, Tanaji; Payghan, Pavan V; Laddha, Kirti; Ghoshal, Nanda; Arvindekar, Akalpita

    2015-08-01

    Rheum emodi is used as a culinary plant across the world and finds an eminent role in the Ayurvedic and traditional Chinese systems of medicine. The plant is known to principally contain 1,8-dihydroxyanthraquinones (DHAQs) like rhein, aloe emodin, emodin, chrysophanol and physcion that possess diverse pharmacological and therapeutic actions. The present work deals with developing a platform technology for isolation of these DHAQs and evaluating their anti-diabetic potential. Herein, we report the anti-hyperglycemic activity and alpha glucosidase (AG) inhibitory actions of five isolated DHAQs from R. emodi. All the five isolated DHAQs showed good anti-hyperglycemic activity with aloe emodin exhibiting maximum lowering of blood glucose in an oral glucose tolerance test. However, on evaluation of the AG inhibitory potential of the DHAQs only emodin exhibited potent intestinal AG inhibition (93 ± 2.16%) with an IC50 notably lower than acarbose. Subsequent kinetic studies indicated a mixed type of inhibition for emodin. In vivo studies using oral maltose load showed almost total inhibition for emodin when compared to acarbose. Molecular docking studies revealed the presence of an allosteric topographically distinct 'quinone binding site' and showed that interaction with Ser 74 occurs exclusively with emodin, which is vital for AG inhibition. The net benefit from the glucose lowering effect and mixed type inhibition by emodin would enable the administration of a small dosage that is safe and non-toxic in the case of prolonged use in treating diabetes.

  9. Purification and biochemical characterization of an alpha-glucosidase from Xanthophyllomyces dendrorhous.

    Science.gov (United States)

    Marín, Dolores; Linde, Dolores; Fernández Lobato, María

    2006-01-30

    Xanthophyllomyces dendrorhous grown in different media shows amylolytic activity, consisting in an extracellular exo-acting enzyme able to hydrolysed alpha,1-4 glycosidic bonds from soluble starch, which also cleaves maltose and malto-oligosaccharides. The enzyme was purified, using basically a couple of chromatography process on DEAE-Sephacel. It is a glycoprotein with a molecular weight estimated to be 60.2 kDa based on its mobility in SDS-PAGE and 115 kDa based on gel filtration. N-linked carbohydrate accounts for 12% of the total mass. It exhibited optimum activity at pH 5.5 and 45 degrees C. Thermostability analysis indicated that it was stable to thermal treatment up to 50 degrees C; 50% of the activity was maintained after 3 h. The rate parameters measured for the hydrolysis of starch and various chain length malto-oligosaccharides shows high catalytic efficiency, calculated by the relationship V(cat)/K(m), for malto-oligosaccharides, such as maltotriose (873 mM(-1) min(-1)), or maltoheptose (698 mM(-1) min(-1)). The new enzyme hydrolysed soluble starch with nearly 3.5- and 1.4-fold lower efficiency than that for maltotriose and maltose, respectively. No activity was found on heterogeneous substrates, such as sucrose and aryl alpha-glucoside, or on isomalto-oligosaccharides. In accordance to substrate specificity profile, the new enzyme was classified as an alpha-glucosidase.

  10. In vitro alpha glucosidase inhibition and free-radical scavenging activity of propolis from Thai stingless bees in mangosteen orchard

    Directory of Open Access Journals (Sweden)

    Boonyadist Vongsak

    2015-10-01

    Full Text Available ABSTRACTThe chemical component and biological activity of propolis depend on flora area of bee collection and bee species. In the study, the propolis from three stingless bee species, Lepidotrigona ventralis Smith, Lepidotrigona terminata Smith, and Tetragonula pagdeni Schwarz, was collected in the same region of mangosteen garden from Thailand. Total phenolic content, alpha glucosidase inhibitory effect, and free-radical scavenging activity using FRAP, ABTS, DPPH assays were determined. The most potent activity of propolis extract was investigated for bioactive compounds and their quantity. The ethanol extract of T. pagdeni propolis had the highest total phenolic content 12.83 ± 0.72 g of gallic acid equivalents in 100 g of the extract, and the strongest alpha glucosidase inhibitory effect with the IC50 of 70.79 ± 6.44 µg/ml. The free-radical scavenging activity evaluated by FRAP, ABTS, DPPH assays showed the FRAP value of 279.70 ± 20.55 µmol FeSO4 equivalent/g extract and the IC50 of 59.52 ± 10.76 and 122.71 ± 11.76 µg/ml, respectively. Gamma- and alpha-mangostin from T. pagdeni propolis extract were isolated and determined for the biological activity. Gamma-mangostin exhibited the strongest activity for both alpha glucosidase inhibitory effect and free-radical scavenging activity. Using HPLC quantitative analysis method, the content of gamma- and alpha-mangostin in the extract was found to be 0.94 ± 0.01 and 2.77 ± 0.08% (w/w, respectively. These findings suggested that T. pagdeni propolis may be used as a more suitable raw material for nutraceutical and pharmaceutical products and these mangostin derivatives as markers.

  11. Twenty-two novel mutations in the lysosomal alpha-glucosidase gene (GAA) underscore the genotype-phenotype correlation in glycogen storage disease type II.

    Science.gov (United States)

    Hermans, Monique M P; van Leenen, Dik; Kroos, Marian A; Beesley, Clare E; Van Der Ploeg, Ans T; Sakuraba, Hitoshi; Wevers, Ron; Kleijer, Wim; Michelakakis, Helen; Kirk, Edwin P; Fletcher, Janice; Bosshard, Nils; Basel-Vanagaite, Lina; Besley, Guy; Reuser, Arnold J J

    2004-01-01

    Patients with glycogen storage disease type II (GSDII, Pompe disease) suffer from progressive muscle weakness due to acid alpha-glucosidase deficiency. The disease is inherited as an autosomal recessive trait with a spectrum of clinical phenotypes. We have investigated 29 cases of GSDII and thereby identified 55 pathogenic mutations of the acid alpha-glucosidase gene (GAA) encoding acid maltase. There were 34 different mutations identified, 22 of which were novel. All of the missense mutations and two other mutations with an unpredictable effect on acid alpha-glucosidase synthesis and function were transiently expressed in COS cells. The effect of a novel splice-site mutation was investigated by real-time PCR analysis. The outcome of our analysis underscores the notion that the clinical phenotype of GSDII is largely dictated by the nature of the mutations in the GAA alleles. This genotype-phenotype correlation makes DNA analysis a valuable tool to help predict the clinical course of the disease.

  12. Functional significance of amylase polymorphism in Drosophila melanogaster. III. Ontogeny of amylase and some alpha-glucosidases.

    Science.gov (United States)

    Hoorn, A J; Scharloo, W

    1980-02-01

    Changes in amylase (E.C. 3.2.1.1), maltase (E.C. 3.2.1.20), sucrase, and PNPGase activities in relation to changes in wet weight and protein content were studied during the development of larvae and adult flies from two strains of Drosophila melanogaster, homozygous for different amylase alleles. All alpha-glucosidase activities increase exponentially during a large part of larval development, parallel to the increase in weight, and drop at the end of the third instar. Amylase activity of the Amy1 strain follows the same pattern. In contrast, amylase activity of the Amy4,6 strain continues its exponential increase longer. In the third larval instar amylase activity in the Amy4,6 strain becomes much higher than in the Amy1 strain. During the first hours of adult life amylase activity of the two strains does not differ. Then Amy4,6 activity starts to rise and becomes much higher (4-5 times) than Amy1 amylase activity, which remains approximately constant. All adult enzyme activities are much higher than in larvae. Comparison of enzyme activity of amylase and alpha-glucosidases in larvae and adults confirms that differences in amylase activities can become important only when starch is a limiting factor in the food.

  13. Structural modeling of mutant alpha-glucosidases resulting in a processing/transport defect in Pompe disease.

    Science.gov (United States)

    Sugawara, Kanako; Saito, Seiji; Sekijima, Masakazu; Ohno, Kazuki; Tajima, Youichi; Kroos, Marian A; Reuser, Arnold J J; Sakuraba, Hitoshi

    2009-06-01

    To elucidate the mechanism underlying transport and processing defects from the viewpoint of enzyme folding, we constructed three-dimensional models of human acid alpha-glucosidase encompassing 27 relevant amino acid substitutions by means of homology modeling. Then, we determined in each separate case the number of affected atoms, the root-mean-square distance value and the solvent-accessible surface area value. The analysis revealed that the amino acid substitutions causing a processing or transport defect responsible for Pompe disease were widely spread over all of the five domains comprising the acid alpha-glucosidase. They were distributed from the core to the surface of the enzyme molecule, and the predicted structural changes varied from large to very small. Among the structural changes, we paid particular attention to G377R and G483R. These two substitutions are predicted to cause electrostatic changes in neighboring small regions on the molecular surface. The quality control system of the endoplasmic reticulum apparently detects these very small structural changes and degrades the mutant enzyme precursor (G377R), but also the cellular sorting system might be misled by these minor changes whereby the precursor is secreted instead of being transported to lysosomes (G483R).

  14. Purification, enzymatic characterization, and nucleotide sequence of a high-isoelectric-point alpha-glucosidase from barley malt

    DEFF Research Database (Denmark)

    Frandsen, T P; Lok, F; Mirgorodskaya, E;

    2000-01-01

    High-isoelectric-point (pI) alpha-glucosidase was purified 7, 300-fold from an extract of barley (Hordeum vulgare) malt by ammonium sulfate fractionation, ion-exchange, and butyl-Sepharose chromatography. The enzyme had high activity toward maltose (k(cat) = 25 s(-1)), with an optimum at pH 4...... in the transition state complex. Mass spectrometry of tryptic fragments assigned the 92-kD protein to a barley cDNA (GenBank accession no. U22450) that appears to encode an alpha-glucosidase. A corresponding sequence (HvAgl97; GenBank accession no. AF118226) was isolated from a genomic phage library using a c......DNA fragment from a barley cDNA library. HvAgl97 encodes a putative 96.6-kD protein of 879 amino acids with 93.8% identity to the protein deduced from U22450. The sequence contains two active site motifs of glycoside hydrolase family 31. Three introns of 86 to 4,286 bp interrupt the coding region. The four...

  15. Enzyme-synthesized highly branched maltodextrins have slow glucose generation at the mucosal alpha-glucosidase level and are slowly digestible "in vivo"

    Science.gov (United States)

    For digestion of starch in humans, alpha-amylase first hydrolyzes starch molecules to produce alpha-limit dextrins, followed by complete hydrolysis to glucose by the mucosal alpha-glucosidases in the small intestine. It is known that alpha-1,6 linkages in starch are hydrolyzed at a lower rate than a...

  16. Transcriptional regulation of the human acid alpha-glucosidase gene. Identification of a repressor element and its transcription factors Hes-1 and YY1.

    Science.gov (United States)

    Yan, B; Heus, J; Lu, N; Nichols, R C; Raben, N; Plotz, P H

    2001-01-19

    Acid alpha-glucosidase, the product of a housekeeping gene, is a lysosomal enzyme that degrades glycogen. A deficiency of this enzyme is responsible for a recessively inherited myopathy and cardiomyopathy, glycogenesis type II. We have previously demonstrated that the human acid alpha-glucosidase gene expression is regulated by a silencer within intron 1, which is located in the 5'-untranslated region. In this study, we have used deletion analysis, electrophoretic mobility shift assay, and footprint analysis to further localize the silencer to a 25-base pair element. The repressive effect on the TK promoter was about 50% in both orientations in expression plasmid, and two transcriptional factors were identified with antibodies binding specifically to the element. Mutagenesis and functional analyses of the element demonstrated that the mammalian homologue 1 of Drosophila hairy and Enhancer of split (Hes-1) binding to an E box (CACGCG) and global transcription factor-YY1 binding to its core site function as a transcriptional repressor. Furthermore, the overexpression of Hes-1 significantly enhanced the repressive effect of the silencer element. The data should be helpful in understanding the expression and regulation of the human acid alpha-glucosidase gene as well as other lysosomal enzyme genes.

  17. Pneumatosis cystoides intestinalis following alpha-glucosidase inhibitor treatment: A case report and review of the literature

    Institute of Scientific and Technical Information of China (English)

    Tatsuhiro Tsujimoto; Hiroshi Fukui; Erika Shioyama; Kei Moriya; Hideto Kawaratani; Yasuyo Shirai; Masahisa Toyohara; Akira Mitoro; Jun-ichi Yarnao; Hisao Fujii

    2008-01-01

    A 69-year-old man was diagnosed as having myasthenia gravis (MG) in September 2004, and treated with thymectomy and prednisolone. He was then diagnosed as having steroid-induced diabetes mellitus, and received sulfonylurea (SU) therapy in May 2005. An alpha-glucosidase inhibitor (αGI) was added in March 2006, resulting in good glycemic control. He experienced symptoms of abdominal distention, increased flatus, and constipation in October 2007, and was admitted into our hospital in late November with hematochezia. Plain abdominal radiography revealed small linear radiolucent clusters in the wall of the colon. Computed tomography (CT) showed intramural air in the sigmoid colon. Colonoscopy revealed multiple smooth surfaced hemispherical protrusions in the sigmoid colon. The diagnosis of pneumatosis cystoides intestinalis (PCI) was made on the basis of these findings. As the αGI voglibose was suspected as the cause of this patient's PCI, treatment was conservative, ceasing voglibose, with fasting and fluid supplementation. The patient progressed well, and was discharged 2 wk later. Recently, several reports of PCI associated with αGI therapy have been published, predominantly in Japan where αGIs are commonly used. If the use of αGIs becomes more widespread, we can expect more reports of this condition on a global scale. The possibility of PCI should be considered in diabetic patients complaining of gastrointestinal symptoms, and the gastrointestinal tract should be thoroughly investigated in these patients.

  18. Alpha-glucosidase inhibitors and hepatotoxicity in type 2 diabetes: a systematic review and meta-analysis

    Science.gov (United States)

    Zhang, Longhao; Chen, Qiyan; Li, Ling; Kwong, Joey S. W.; Jia, Pengli; Zhao, Pujing; Wang, Wen; Zhou, Xu; Zhang, Mingming; Sun, Xin

    2016-09-01

    Alpha-glucosidase inhibitors (AGIs) was reported to be associated with several rare adverse hepatic events, but with inconsistent results. We aimed to investigate the risk of hepatotoxicity associated with the use of AGIs in patients with type 2 diabetes mellitus (T2DM), and performed a systematic review and meta-analysis. Fourteen studies (n = 2881) were eligible, all of which were RCTs. Meta-analysis of data regarding elevation of more than 3-fold the upper limit of normal (ULN) of AST and ALT showed statistically significant differences between AGIs treatment versus control (OR 6.86, 95% CI 2.50 to 18.80; OR 6.48, 95% CI 2.40 to 17.49). Subgroup analyses of elevation of more than 1.8-fold ULN of AST and ALT by dose of AGIs showed differential effects on AST and ALT (AST: OR 0.38 vs 7.31, interaction P = 0.003 ALT: OR 0.32 vs 4.55, interaction p = 0.02). Meta-analysis showed that AGIs might increase the risk of hepatotoxicity, and higher dose appeared to be associated with higher risk of hepatotoxicity. However, the evidence is limited with surrogate measures (i.e. ALT and AST), and no clinically important adverse events were observed.

  19. Miconia sp. Increases mRNA Levels of PPAR Gamma and Inhibits Alpha Amylase and Alpha Glucosidase

    Science.gov (United States)

    Ortíz-Martinez, David Mizael; Rivas-Morales, Catalina; de la Garza-Ramos, Myriam Angelica; Verde-Star, Maria Julia; Nuñez-Gonzalez, Maria Adriana

    2016-01-01

    Diabetes mellitus is a public health problem worldwide. For this reason, ethanolic extract of Miconia sp. from Oaxaca, Mexico, was selected in search of an alternative against this disease. The effect of Miconia sp. on mRNA expression of PPARγ on cell line 3T3-L1, its effect on alpha amylase and alpha glucosidase, lipid accumulation during adipogenesis, and cell viability on VERO cells were evaluated. The mRNA levels of PPARγ increased on 1.393 ± 0.008 folds, lipid accumulation was increased by 29.55% with Miconia sp. extract and 34.57% with rosiglitazone, and α-amylase and α-glycosidase were inhibited with IC50 values from 28.23 ± 2.15 μg/mL and 1.95 ± 0.15 μg/mL, respectively; the IC50 on antiproliferative activity on VERO cells was 314.54 ± 45.40 μg/mL. In case of α-amylase and α-glycosidase assays, IC50 (inhibitory concentration 50) refers to necessary extract amounts to inhibit 50% of enzymatic activity. On the other hand, on antiproliferative activity, IC50 (inhibitory concentration 50) refers to necessary extract amounts to inhibit 50% of cell proliferation. It was concluded that the compounds present in Miconia sp. ethanolic extract increase mRNA expression of PPARγ, inhibit α-amylase and α-glucosidase, and increase lipid accumulation. It constitutes an alternative as adjuvant in diabetes mellitus treatment; therefore, we recommend continuing identifying the compounds responsible for its promising in vivo antidiabetic activity. PMID:27478477

  20. Miconia sp. Increases mRNA Levels of PPAR Gamma and Inhibits Alpha Amylase and Alpha Glucosidase.

    Science.gov (United States)

    Ortíz-Martinez, David Mizael; Rivas-Morales, Catalina; de la Garza-Ramos, Myriam Angelica; Verde-Star, Maria Julia; Nuñez-Gonzalez, Maria Adriana; Leos-Rivas, Catalina

    2016-01-01

    Diabetes mellitus is a public health problem worldwide. For this reason, ethanolic extract of Miconia sp. from Oaxaca, Mexico, was selected in search of an alternative against this disease. The effect of Miconia sp. on mRNA expression of PPARγ on cell line 3T3-L1, its effect on alpha amylase and alpha glucosidase, lipid accumulation during adipogenesis, and cell viability on VERO cells were evaluated. The mRNA levels of PPARγ increased on 1.393 ± 0.008 folds, lipid accumulation was increased by 29.55% with Miconia sp. extract and 34.57% with rosiglitazone, and α-amylase and α-glycosidase were inhibited with IC50 values from 28.23 ± 2.15 μg/mL and 1.95 ± 0.15 μg/mL, respectively; the IC50 on antiproliferative activity on VERO cells was 314.54 ± 45.40 μg/mL. In case of α-amylase and α-glycosidase assays, IC50 (inhibitory concentration 50) refers to necessary extract amounts to inhibit 50% of enzymatic activity. On the other hand, on antiproliferative activity, IC50 (inhibitory concentration 50) refers to necessary extract amounts to inhibit 50% of cell proliferation. It was concluded that the compounds present in Miconia sp. ethanolic extract increase mRNA expression of PPARγ, inhibit α-amylase and α-glucosidase, and increase lipid accumulation. It constitutes an alternative as adjuvant in diabetes mellitus treatment; therefore, we recommend continuing identifying the compounds responsible for its promising in vivo antidiabetic activity.

  1. Miconia sp. Increases mRNA Levels of PPAR Gamma and Inhibits Alpha Amylase and Alpha Glucosidase

    Directory of Open Access Journals (Sweden)

    David Mizael Ortíz-Martinez

    2016-01-01

    Full Text Available Diabetes mellitus is a public health problem worldwide. For this reason, ethanolic extract of Miconia sp. from Oaxaca, Mexico, was selected in search of an alternative against this disease. The effect of Miconia sp. on mRNA expression of PPARγ on cell line 3T3-L1, its effect on alpha amylase and alpha glucosidase, lipid accumulation during adipogenesis, and cell viability on VERO cells were evaluated. The mRNA levels of PPARγ increased on 1.393±0.008 folds, lipid accumulation was increased by 29.55% with Miconia sp. extract and 34.57% with rosiglitazone, and α-amylase and α-glycosidase were inhibited with IC50 values from 28.23±2.15 μg/mL and 1.95±0.15 μg/mL, respectively; the IC50 on antiproliferative activity on VERO cells was 314.54±45.40 μg/mL. In case of α-amylase and α-glycosidase assays, IC50 (inhibitory concentration 50 refers to necessary extract amounts to inhibit 50% of enzymatic activity. On the other hand, on antiproliferative activity, IC50 (inhibitory concentration 50 refers to necessary extract amounts to inhibit 50% of cell proliferation. It was concluded that the compounds present in Miconia sp. ethanolic extract increase mRNA expression of PPARγ, inhibit α-amylase and α-glucosidase, and increase lipid accumulation. It constitutes an alternative as adjuvant in diabetes mellitus treatment; therefore, we recommend continuing identifying the compounds responsible for its promising in vivo antidiabetic activity.

  2. N-glycans of recombinant human acid alpha-glucosidase expressed in the milk of transgenic rabbits.

    Science.gov (United States)

    Jongen, Susanne P; Gerwig, Gerrit J; Leeflang, Bas R; Koles, Kate; Mannesse, Maurice L M; van Berkel, Patrick H C; Pieper, Frank R; Kroos, Marian A; Reuser, Arnold J J; Zhou, Qun; Jin, Xiaoying; Zhang, Kate; Edmunds, Tim; Kamerling, Johannis P

    2007-06-01

    Pompe disease is a lysosomal glycogen storage disorder characterized by acid alpha-glucosidase (GAA) deficiency. More than 110 different pathogenic mutations in the gene encoding GAA have been observed. Patients with this disease are being treated by intravenous injection of recombinant forms of the enzyme. Focusing on recombinant approaches to produce the enzyme means that specific attention has to be paid to the generated glycosylation patterns. Here, human GAA was expressed in the mammary gland of transgenic rabbits. The N-linked glycans of recombinant human GAA (rhAGLU), isolated from the rabbit milk, were released by peptide-N(4)-(N-acetyl-beta-glucosaminyl)asparagine amidase F. The N-glycan pool was fractionated and purified into individual components by a combination of anion-exchange, normal-phase, and Sambucus nigra agglutinin-affinity chromatography. The structures of the components were analyzed by 500 MHz one-dimensional and 600 MHz cryo two-dimensional (total correlation spectroscopy [TOCSY] nuclear Overhauser enhancement spectroscopy) (1)H nuclear magnetic resonance spectroscopy, combined with two-dimensional (31)P-filtered (1)H-(1)H TOCSY spectroscopy, matrix-assisted laser desorption ionization time-of-flight mass spectrometry, and high-performance liquid chromatography (HPLC)-profiling of 2-aminobenzamide-labeled glycans combined with exoglycosidase digestions. The recombinant rabbit glycoprotein contained a broad array of different N-glycans, comprising oligomannose-, hybrid-, and complex-type structures. Part of the oligomannose-type glycans showed the presence of phospho-diester-bridged N-acetylglucosamine. For the complex-type glycans (partially) (alpha2-6)-sialylated (nearly only N-acetylneuraminic acid) diantennary structures were found; part of the structures were (alpha1-6)-core-fucosylated or (alpha1-3)-fucosylated in the upper antenna (Lewis x). Using HPLC-mass spectrometry of glycopeptides, information was generated with respect to the

  3. Chemical chaperones improve transport and enhance stability of mutant alpha-glucosidases in glycogen storage disease type II.

    Science.gov (United States)

    Okumiya, Toshika; Kroos, Marian A; Vliet, Laura Van; Takeuchi, Hiroaki; Van der Ploeg, Ans T; Reuser, Arnold J J

    2007-01-01

    Glycogen storage disease type II (GSDII; Pompe disease or acid maltase deficiency) is an autosomal recessive disorder caused by lysosomal acid alpha-glucosidase (AalphaGlu) deficiency and manifests predominantly as skeletal muscle weakness. Defects in post-translational modification and transport of mutant AalphaGlu species are frequently encountered and may potentially be corrected with chaperone-mediated therapy. In the present study, we have tested this hypothesis by using deoxynojirimycin and derivatives as chemical chaperones to correct the AalphaGlu deficiency in cultured fibroblasts from patients with GSDII. Four mutant phenotypes were chosen: Y455F/Y455F, P545L/P545L, 525del/R600C and D645E/R854X. In case of Y455F/Y455F and P545L/P545L, N-(n-butyl)deoxynojirimycin (NB-DNJ) restored the transport, maturation and activity of AalphaGlu in a dose dependent manner, while it had no effect on the reference enzyme beta-hexosaminidase. NB-DNJ promoted export from the endoplasmic reticulum (ER) to the lysosomes and stabilized the activity of mutant AalphaGlu species, Y455F and P545L, inside the lysosomes. In long-term culture, the AalphaGlu activity in the fibroblasts from the patients with mutant phenotypes, Y455F/Y455F and P545L/P545L, increased up to 14.0- and 7.9-fold, respectively, in the presence of 10mumol/L NB-DNJ. However, the effect of NB-DNJ on Y455F/Y455F subsided quickly after removal of the compound. We conclude that NB-DNJ acts in low concentration as chemical chaperone for certain mutant forms of AalphaGlu that are trapped in the ER, poorly transported or labile in the lysosomal environment. Chemical chaperone therapy could create new perspectives for therapeutic intervention in GSDII.

  4. Production of enzymatically active recombinant full-length barley high pI alpha-glucosidase of glycoside family 31 by high cell-density fermentation of Pichia pastoris and affinity purification

    DEFF Research Database (Denmark)

    Næsted, Henrik; Kramhøft, Birte; Lok, F.

    2006-01-01

    of the alcohol oxidase 1 promoter using methanol induction of P. pastoris fermentation in a Biostat B 5 L reactor. Forty-two milligrams a-glucosidase was purified from 3.5 L culture in four steps applying an N-terminal hexa-histidine tag. The apparent molecular mass of the recombinant alpha-glucosidase was 100 k...

  5. Alternative Agents in Type 1 Diabetes in Addition to Insulin Therapy: Metformin, Alpha-Glucosidase Inhibitors, Pioglitazone, GLP-1 Agonists, DPP-IV Inhibitors, and SGLT-2 Inhibitors.

    Science.gov (United States)

    DeGeeter, Michelle; Williamson, Bobbie

    2016-04-01

    Insulin is the mainstay of current treatment for patients with type 1 diabetes mellitus (T1DM). Due to increasing insulin resistance, insulin doses are often continually increased, which may result in weight gain for patients. Medications currently approved for the treatment of type 2 diabetes offer varying mechanisms of action that can help to reduce insulin resistance and prevent or deter weight gain. A MEDLINE search was conducted to review literature evaluating the use of metformin, alpha-glucosidase inhibitors, pioglitazone, glucagon-like peptide 1 agonists, dipeptidyl peptidase, and sodium-dependent glucose transporter 2 inhibitors, in patients with T1DM. Varying results were found with some benefits including reductions in hemoglobin A1c, decreased insulin doses, and favorable effects on weight. Of significance, a common fear of utilizing multiple therapies for diabetes treatment is the risk of hypoglycemia, and this review displayed limited evidence of hypoglycemia with multiple agents.

  6. Hes-1, a known transcriptional repressor, acts as a transcriptional activator for the human acid alpha-glucosidase gene in human fibroblast cells.

    Science.gov (United States)

    Yan, Bo; Raben, Nina; Plotz, Paul H

    2002-03-01

    Hes-1, the mammalian homologue 1 of Drosophila hairy and Enhancer of split proteins, belongs to a family of basic helix-loop-helix proteins that are essential to neurogenesis, myogenesis, hematopoiesis, and sex determination. Hes-1 is a transcriptional repressor for a number of known genes including the human acid alpha-glucosidase (GAA) gene as we have previously shown in Hep G2 cells. The human GAA gene encodes the enzyme for glycogen breakdown in lysosomes, deficiency of which results in Glycogen Storage Disease type II (Pompe syndrome). Using constructs containing the DNA element that demonstrates repressive activity in Hep G2 cells and conditions in which the same transcription factors, Hes-1 and YY1, bind, we have shown that this element functions as an enhancer in human fibroblasts. Site-directed mutagenesis and overexpression of Hes-1 showed that Hes-1 functions as a transcriptional activator. The dual function of Hes-1 we have found is likely to contribute to the subtle tissue-specific control of this housekeeping gene.

  7. Enzyme-assisted extraction of phenolics from winemaking by-products: Antioxidant potential and inhibition of alpha-glucosidase and lipase activities.

    Science.gov (United States)

    de Camargo, Adriano Costa; Regitano-d'Arce, Marisa Aparecida Bismara; Biasoto, Aline Camarão Telles; Shahidi, Fereidoon

    2016-12-01

    Phenolics in food and agricultural processing by-products exist in the soluble and insoluble-bound forms. The ability of selected enzymes in improving the extraction of insoluble-bound phenolics from the starting material (experiment I) or the residues containing insoluble-bound phenolics (experiment II) were evaluated. Pronase and Viscozyme improved the extraction of insoluble-bound phenolics as evaluated by total phenolic content, antioxidant potential as determined by ABTS and DPPH assays, and hydroxyl radical scavenging capacity, reducing power as well as evaluation of inhibition of alpha-glucosidase and lipase activities. Viscozyme released higher amounts of gallic acid, catechin, and prodelphinidin dimer A compared to Pronase treatment. Furthermore, p-coumaric and caffeic acids, as well as procyanidin dimer B, were extracted with Viscozyme but not with Pronase treatment. Solubility plays an important role in the bioavailability of phenolic compounds, hence this study may assist in better exploitation of phenolics from winemaking by-products as functional food ingredients and/or supplements.

  8. Alpha-glucosidase inhibitor, acarbose, improves glycamic control and reduces body weight in type 2 diabetes: Findings on indian patients from the pooled data analysis

    Directory of Open Access Journals (Sweden)

    Sanjay Kalra

    2013-01-01

    Full Text Available Alpha-glucosidase inhibitors are widely used especially in Asian countries as a treatment option for type 2 diabetes patients with high postprandial glycemia (PPG. The higher carbohydrate in the Indian diets lead to greater prandial glycemic excursion, increased glucosidase, and incretin activity in the gut and may need special therapeutic strategies to tackle these glucose peaks. This is the subgroup analysis of Indian subjects who participated in the GlucoVIP study that investigated the effectiveness and tolerability of acarbose as add-on or monotherapy in a range of patients with type 2 diabetes mellitus. A total of 1996 Indian patients were included in the effectiveness analysis. After 12.5 weeks (mean, the mean change in 2-hour PPG from baseline was −74.4 mg/dl, mean HbA1c decreased by -1.0%, and mean fasting blood glucose decreased by -37.9 mg/dl. The efficacy of acarbose was rated "very good" or "good" in 91.1% of patients, and tolerability as "very good" or "good" in 88.0% of patients. The results of this observational study suggest that acarbose was effective and well tolerated in the Indian patients with T2DM.

  9. KAPASITAS ANTIOKSIDAN DAN INHIBITOR ALFA GLUKOSIDASE EKSTRAK UMBI BAWANG DAYAK [Antioxidant and Alpha-Glucosidase Inhibitory Properties of Bawang Dayak Bulb Extracts

    Directory of Open Access Journals (Sweden)

    Andi Early Febrinda*

    2013-12-01

    Full Text Available Bawang dayak (Eleutherine palmifolia is an indigenous plant in Borneo traditionally used by Dayak tribes to treat any kind of degenerative deseases including diabetes mellitus. The purpose of this research was to measure antioxidant and antidiabetic capacities of water and ethanolic extracts of bawang dayak bulb. Parameters evaluated in this research were phytochemical screening, total phenolics, flavonoid content, DPPH free-radical scavenging activity, and alpha glucosidase inhibiting (AGI activity. The result showed that the total phenolics and flavonoid content in bawang dayak ethanolic extract (217.71 mg GAE/g and 65.35 mg QE/g were higher than that of the water extract (139.93 mg GAE/g and 16.95 mg QE/g. The ethanolic extract also had higher antioxidant and AGI activities (IC50 112 and 241 ppm than that of the water extract (IC50 526 and 505 ppm. In addition, the IC50 values for AGI in bawang dayak ethanolic extract was lower than acarbose which is known as a commercial antidiabetic agent.

  10. High frequency of acid alpha-glucosidase pseudodeficiency complicates newborn screening for glycogen storage disease type II in the Japanese population.

    Science.gov (United States)

    Kumamoto, Shingo; Katafuchi, Tatsuya; Nakamura, Kimitoshi; Endo, Fumio; Oda, Eri; Okuyama, Torayuki; Kroos, Marian A; Reuser, Arnold J J; Okumiya, Toshika

    2009-07-01

    To investigate the feasibility of newborn screening for glycogen storage disease type II (GSDII; Pompe disease or acid maltase deficiency) in the Japanese population, we assayed the acid alpha-glucosidase activity in dried blood spots from 715 Japanese newborns and 18 previously diagnosed patients using a fluorometric procedure. The enzyme activity of apparently healthy newborns showed a bimodal distribution. The median activity of the minor group (31 individuals, 4.3% of the samples) was 6.5 times lower than that of the major group. Four of the 715 control samples (0.56%) fell in the patient range. We then analyzed genomic DNA, extracted from the same blood spots, for the occurrence of two sequence variants, c.1726G>A and c.2065G>A, known to cause "pseudodeficiency". This analysis revealed that 27 of 28 individuals homozygous for c.[1726A; 2065A] belonged to the minor group. One c.[1726A; 2065A] homozygote had just slightly higher activity. Twelve of the 18 patients with GSDII either had one (9 cases) or two (3 cases) c.[1726A; 2065A] alleles. The frequency of this allele was double in the patient compared to the control group (0.42 vs 0.19) at the expense of a lower frequency of the c.[1726G; 2065G] and c.[1726G; 2065A] alleles (0.58 vs 0.71 and 0 vs 0.1). These findings illustrate that c.[1726A; 2065A] homozygosity among apparently healthy individuals (3.9 per 100) complicates newborn screening for GSDII in Japan, and further that one or more pathogenic mutations are associated with the c.[1726A; 2065A] allele.

  11. Optimization of Inhibitory Reaction System of Folium Mori Extracts on Alpha-glucosidase Activity and Enzyme Kinetics%桑叶提取物抑制α-葡萄糖苷酶活性体系的优化及动力学研究

    Institute of Scientific and Technical Information of China (English)

    吕欢; 罗明琍; 方飞; 吴新荣

    2012-01-01

    Objective To establish an optimized inhibitory reaction system for the research of Folium Mori extracts in inhibiting alpha-glucosidase activity and to investigate the enzyme kinetics. Methods We studied the inhibitive effects of Folium Mori extracts on alpha-glucosidase by using enzyme-inhibitor model and optimized the reaction system by optimizing the substrate concentration, the amount of enzyme and the reaction time. We also investigated the inhibitory kinetic characteristics of extracts by using Lineweaver-Burk method. Results The optimized reaction system consisted of 40 mg/mL of the substrate concentration, 0.2 mL of enzyme and 25 min for reaction time. Inhibitory kinetic results indicated that the extracts were mixed competitive inhibitors, but the positive drug was noncompetitive inhibitor. Conclusion Folium Mori extracts have inhibitory effect on a- glucosidase activity, and the inhibitory type is in mixed competitive inhibition, which are expected to be used to develop as anti-diabetes drugs.%目的 研究桑叶提取物抑制α-葡萄糖苷酶活性反应体系的条件优化及酶动力学.方法 采用酶-抑制剂模型法,优化底物浓度、酶用量和反应时间,确定最佳反应体系,研究桑叶提取物对α-葡萄糖苷酶的抑制作用,并采用双倒数作图法研究桑叶提取物的酶抑制动力学特性.结果 当蔗糖浓度为40 mg/mL,酶用量为0.2mL(约1.6 U/mL),反应时间为25 min时为最佳反应体系;酶抑制动力学实验表明,桑叶提取物的抑制类型为混合型竞争性抑制,阳性对照为非竞争性抑制.结论 桑叶提取物对α-葡萄糖苷酶活性有抑制作用;桑叶提取物对α-葡萄糖苷酶抑制活性为混合性竞争性抑制,可以用于开发降糖药物.

  12. Relato do primeiro paciente brasileiro com a forma infantil da doença de Pompe tratado com alfa-glicosidase recombinante humana Report of the first Brazilian infantile Pompe disease patient to be treated with recombinant human acid alpha-glucosidase

    Directory of Open Access Journals (Sweden)

    Sandra J. Pereira

    2008-06-01

    Full Text Available Objetivo: Relatar o primeiro caso de forma infantil da doença de Pompe tratado no Brasil. Descrição: Trata-se de doença de depósito lisossomal que se caracteriza por defeitos da enzima alfa-glicosidase ácida, com acúmulo intracelular de glicogênio, principalmente nos músculos. São descritas a forma infantil e tardia. Desde 2006, está disponível tratamento com enzima recombinante humana. Descreve-se o primeiro caso de forma infantil da doença tratado no Brasil. Trata-se de menina com 2,5 meses de idade e progressão rápida da doença, com perda dos movimentos dos membros, miocardiopatia hipertrófica e insuficiência respiratória aos 7 meses de idade. Após 10 meses de tratamento, apresentou boa resposta clínica, com remissão da insuficiência respiratória, recuperação parcial dos movimentos dos membros e melhora importante do quadro cardiológico. Comentários: Apesar de pouco freqüente, a forma infantil da doença de Pompe é letal. A disponibilidade de tratamento eficaz aumenta a necessidade de conhecimento e diagnóstico precoce da doença.Objective: To describe the first case of infantile Pompe disease to be treated in Brazil. Description: Pompe disease is a glycogen storage disease related to defects in the acid alpha-glucosidase enzyme, leading to an intracellular accumulation of glycogen, mainly in muscles. Two forms are described: infantile and juvenile. Since 2006, treatment with recombinant human acid alpha-glucosidase has been available. This article describes the first case of infantile Pompe disease treated in Brazil. A girl presented at 2.5 months of age with rapid disease progression, exhibiting severe hypotonia, loss of movements in both upper and lower limbs and hypertrophic cardiomyopathy, progressing to respiratory failure by the age of 7 months. After 10 months of treatment, she exhibited a good clinical response, with remission of the respiratory failure, partial recovery of arm and leg movements and

  13. Biogenesis of lysosomal enzymes in the alpha-glucosidase II-deficient modA mutant of Dictyostelium discoideum: retention of alpha-1,3-linked glucose on N-linked oligosaccharides delays intracellular transport but does not alter sorting of alpha-mannosidase or beta-glucosidase.

    Science.gov (United States)

    Ebert, D L; Bush, J M; Dimond, R L; Cardelli, J A

    1989-09-01

    The endoplasmic reticulum-localized enzyme alpha-glucosidase II is responsible for removing the two alpha-1,3-linked glucose residues from N-linked oligosaccharides of glycoproteins. This activity is missing in the modA mutant strain, M31, of Dictyostelium discoideum. Results from both radiolabeled pulse-chase and subcellular fractionation experiments indicate that this deficiency did not prevent intracellular transport and proteolytic processing of the lysosomal enzymes, alpha-mannosidase and beta-glucosidase. However, the rate at which the glucosylated precursors left the rough endoplasmic reticulum was several-fold slower than the rate at which the wild-type precursors left this compartment. Retention of glucose residues did not disrupt the binding of the precursor forms of the enzymes with intracellular membranes, indicating that the delay in movement of proteins from the ER did not result from lack of association with membranes. However, the mutant alpha-mannosidase precursor contained more trypsin-sensitive sites than did the wild-type precursor, suggesting that improper folding of precursor molecules might account for the slow rate of transport to the Golgi complex. Percoll density gradient fractionation of extracts prepared from M31 cells indicated that the proteolytically processed mature forms of alpha-mannosidase and beta-glucosidase were localized to lysosomes. Finally, the mutation in M31 may have other, more dramatic, effects on the lysosomal system since two enzymes, N-acetylglucosaminidase and acid phosphatase, were secreted much less efficiently from lysosomal compartments by the mutant strain.

  14. Study on the Optimum Proportion of Radix Astragalus with Flos Carthami on alpha - Glucosidase and Antioxidant Activities%黄芪与红花抑制α-葡萄糖苷酶及抗氧化活性的最佳配比研究

    Institute of Scientific and Technical Information of China (English)

    廖晖; 王孝敏

    2015-01-01

    目的:依照国家食品药品监督管理总局(CFDA)制定的黄芪注射液与红花注射液的制备方法,分别对黄芪与红花进行相关提取。研究黄芪、红花提取物抑制α-葡萄糖苷酶及抗氧化活性的最佳配比。方法液相色谱仪测定黄芪中黄芪甲苷的含量,以山奈酚为标准品测定红花中总黄酮的含量。以4-甲基-伞形酮-β- D -半乳糖苷(4- MUG)为反应底物,测定样品对酵母α-葡萄糖苷酶活性的影响,阿卡波糖为阳性对照。以氧自由基清除能( Oxygen Radical Absorbance Capacity, ORAC)法测定样品的抗氧化活性,维生素 C 为阳性对照。结果每1 ml 黄芪提取液中含有0.127 mg黄芪甲苷,每1 ml 红花提取液中含0.57 mg 总黄酮,均符合 CFDA 有关活性成分的含量要求。红花抑制α-葡萄糖苷酶活性的半数抑制浓度(IC50)为(32.8±5.7)μg/ ml,显著低于黄芪[(1686±810)μg/ ml,P <0.01];黄芪与红花生药比为10:1、5:1、2:1时的 IC50值均显著低于20:1时(P <0.01)。红花的 ORAC 值为(1090±161)μmol TE/ g,显著高于黄芪[(205±32)μmol TE/ g,P <0.01];生药比为5:1及2:1时的 ORAC 值分别显著高于20:1及 10:1时(P <0.01)。结论黄芪与红花均具有抑制酵母α-葡萄糖苷酶活性的作用及一定的抗氧化活性,在同时改善以上两项指标方面,生药比为5:1及2:1时较20:1时作用显著(P <0.01)。%Objective To prepare radix astragalus injection and flos carthami injection according to the method of the China Food and Drug Administration(CFDA)and extract them respectively so as to study the optimum proportion of them on alpha - glucosidase and antioxidant activities. Methods The liquid chro-matograph was used to determine the content of astragaloside in radix astragalus. Kaempferol was used as the standard to determine the content of general flavone in flos carthami. 4 - MUG was as the

  15. Alpha-glucosidase inhibitory effect and inorganic constituents of Phyllanthus amarus Schum. & Thonn. ash

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    Malinee Wongnawa

    2014-10-01

    Full Text Available This study investigated the -glucosidase inhibitory effect and determined the concentration of some inorganic constituents in P. amarus ash. Oral glucose and sucrose tolerance test were performed on normal mice. In vitro -glucosidase inhibitory activity was evaluated by using yeast a-glucosidase. The element concentrations were measured by inductively coupled plasma (ICP spectroscopy. Single oral administration of P. amarus ash did not show antihyperglycemic effect after glucose administration, but decreased blood glucose level after sucrose administration. The ash showed -glucosidase inhibitory activity in vitro with IC50 of 982 mg/mL. The concentrations of K, Ca, Mg, Mn, Fe, Zn, Cu, Pb, Cr, Ni and Co in P. amarus ash were 35049.80±340.64, 3337.24±52.10, 1368.52±13.29, 90.81±1.34, 87.68±1.15, 18.28±0.22, 4.69±0.07, 1.07±0.15, 0.29±0.03, 0.20±0.04 and 0.10±0.02 mg/g, respectively. These results indicate that the antihyperglycemic effect of P. amarus ash might be partly due to the -glucosidase inhibitory activity of the inorganic constituents.

  16. Long-term intravenous treatment of Pompe disease with recombinant human alpha-glucosidase from milk

    NARCIS (Netherlands)

    J.M.P. van den Hout (Johanna); B. Sibbles (Barbara); J.P. Brakenhoff (Just); A.H. Cromme-Dijkhuis (Adri); N. Weisglas-Kuperus (Nynke); A.J.J. Reuser (Arnold); M.A. Boer (Marijke); J.A.M. Smeitink (Jan); O.P. van Diggelen (Otto); E. Voort (Edwin); E.J.J.M. van Corven (Emiel); H. van Hirtum (Hans); J.H.J. Kamphoven (Joep); A.T. van der Ploeg (Ans); J. van Hove (Johan); W.F.M. Arts (Willem Frans); P.A. van Doorn (Pieter); J.B.C. de Klerk (Johannes); M.C.B. Loonen (Christa); A.G. Vulto (Arnold); M.A. Kroos (Marian); W.C.J. Hop (Wim); L.P.F. Winkel (Léon); G. de Jong (Gerard)

    2004-01-01

    textabstractOBJECTIVE: Recent reports warn that the worldwide cell culture capacity is insufficient to fulfill the increasing demand for human protein drugs. Production in milk of transgenic animals is an attractive alternative. Kilogram quantities of product per year can be obtain

  17. Long-term intravenous treatment of Pompe disease with recombinant human alpha-glucosidase from milk.

    NARCIS (Netherlands)

    Hout, J.M. van den; Kamphoven, J.H.; Winkel, L.P.; Arts, W.F.M.; Klerk, J.B.C. de; Loonen, M.C.B.; Vulto, A.G.; Cromme-Dijkhuis, A.H.; Weisglas-Kuperus, N.; Hop, W.C.J.; Hirtum, H. van; Diggelen, O.P. van; Boer, M. de; Kroos, M.A.; Doorn, P.A. van; Voort, E.I. van der; Sibbles, B.; Corven, E.J. van; Brakenhoff, J.P.; Hove, J.L. van; Smeitink, J.A.M.; Jong, G. de; Reuser, A.J.J.; Ploeg, A.T. van der

    2004-01-01

    OBJECTIVE: Recent reports warn that the worldwide cell culture capacity is insufficient to fulfill the increasing demand for human protein drugs. Production in milk of transgenic animals is an attractive alternative. Kilogram quantities of product per year can be obtained at relatively low costs, ev

  18. Antiviral therapies targeting host ER alpha-glucosidases: current status and future directions.

    Science.gov (United States)

    Chang, Jinhong; Block, Timothy M; Guo, Ju-Tao

    2013-09-01

    Endoplasmic reticulum (ER)-resident α-glucosidases I and II sequentially trim the three terminal glucose moieties on N-linked glycans attached to nascent glycoproteins. These reactions are the first steps of N-linked glycan processing and are essential for proper folding and function of many glycoproteins. Because most viral envelope glycoproteins contain N-linked glycans, inhibition of ER α-glucosidases with derivatives of 1-deoxynojirimycin (DNJ) or castanospermine (CAST), two well-studied pharmacophores of α-glucosidase inhibitors, efficiently disrupts the morphogenesis of a broad spectrum of enveloped viruses. Moreover, both DNJ and CAST derivatives have been demonstrated to prevent the death of mice infected with several distinct flaviviruses and filoviruses and suppress the multiplication of several other species of viruses in infected animals. N-Butyl derivative of DNJ (NB-DNJ) and 6 O-bytanoyl prodrug of CAST (Bu-CAST) have been evaluated in human clinical trials for their antiviral activities against human immunodeficiency virus and hepatitis C virus, and there is an ongoing trial of treating dengue patients with Bu-CAST. This article summarizes the current status of ER α-glucosidase-targeted antiviral therapy and proposes strategies for development of more efficacious and specific ER α-glucosidase inhibitors as broad-spectrum, drug resistance-refractory antiviral therapeutics. These host function-targeted, broad-spectrum antiviral agents do not rely on time-consuming etiologic diagnosis, and should therefore be particularly promising in the management of viral hemorrhagic fever and respiratory tract viral infections, medical conditions that can be caused by many different enveloped RNA viruses, with a short window for medical intervention.

  19. Bergamotane Sesquiterpenes with Alpha-Glucosidase Inhibitory Activity from the Plant Pathogenic Fungus Penicillium expansum.

    Science.gov (United States)

    Ying, You-Min; Fang, Cheng-An; Yao, Feng-Qi; Yu, Yuan; Shen, Ying; Hou, Zhuo-Ni; Wang, Zhen; Zhang, Wei; Shan, Wei-Guang; Zhan, Zha-Jun

    2017-01-01

    Two new bergamotane sesquiterpene lactones, named expansolides C and D (1 and 2), together with two known compounds expansolides A and B (3 and 4), were isolated from the plant pathogenic fungus Penicillium expansum ACCC37275. The structures of the new compounds were established by detailed analyses of the spectroscopic data, especially 1D-, 2D-NMR, and HR-ESI-MS. In an in vitro bioassay, the epimeric mixture of expansolides C and D (1 and 2) (in a ratio of 2:1 at the temprature of the bioassay) exhibited more potent α-glucosidase inhibitory activity (IC50 =0.50 ± 0.02 mm) as compared with the positive control acarbose (IC50 = 1.90 ± 0.05 mm). To the best of our knowledge, it was the first report on the α-glucosidase inhibitory activity of bergamotane sesquiterpenes.

  20. Alpha-Glucosidase Enzyme Biosensor for the Electrochemical Measurement of Antidiabetic Potential of Medicinal Plants

    Science.gov (United States)

    Mohiuddin, M.; Arbain, D.; Islam, A. K. M. Shafiqul; Ahmad, M. S.; Ahmad, M. N.

    2016-02-01

    A biosensor for measuring the antidiabetic potential of medicinal plants was developed by covalent immobilization of α-glucosidase (AG) enzyme onto amine-functionalized multi-walled carbon nanotubes (MWCNTs-NH2). The immobilized enzyme was entrapped in freeze-thawed polyvinyl alcohol (PVA) together with p-nitrophenyl-α- d-glucopyranoside (PNPG) on the screen-printed carbon electrode at low pH to prevent the premature reaction between PNPG and AG enzyme. The enzymatic reaction within the biosensor is inhibited by bioactive compounds in the medicinal plant extracts. The capability of medicinal plants to inhibit the AG enzyme on the electrode correlates to the potential of the medicinal plants to inhibit the production of glucose from the carbohydrate in the human body. Thus, the inhibition indicates the antidiabetic potential of the medicinal plants. The performance of the biosensor was evaluated to measure the antidiabetic potential of three medicinal plants such as Tebengau ( Ehretis laevis), Cemumar ( Micromelum pubescens), and Kedondong ( Spondias dulcis) and acarbose (commercial antidiabetic drug) via cyclic voltammetry, amperometry, and spectrophotometry. The cyclic voltammetry (CV) response for the inhibition of the AG enzyme activity by Tebengau plant extracts showed a linear relation in the range from 0.423-8.29 μA, and the inhibition detection limit was 0.253 μA. The biosensor exhibited good sensitivity (0.422 μA/mg Tebengau plant extracts) and rapid response (22 s). The biosensor retains approximately 82.16 % of its initial activity even after 30 days of storage at 4 °C.

  1. Synthesis and Characterization of New Chromogenic Substrates for Exoglycosidases : alpha-Glucosidase, alpha-Mannosidase, and beta-Galactosidase

    OpenAIRE

    Iga, Dumitru Petru; Schmidt, Richard; Iga, Silvia; Hotoleanu, Corina Loredana; Duica, Florentina; Nicolescu, Alina; Gitman, Silvia Stefania

    2013-01-01

    Glycosides of 4-nitrocatechol (1,2-dihydroxy 4-nitrobenzene) with α-D-glucopyranose and α-D-mannopyranose were synthesized by the glycosylation of phenol with peracetylated sugars in the presence of BF3·OBu2 . The glycoside of 4-nitrocatechol with β-D-galactopyranose was prepared by the glycosylation of this phenol as sodium phenoxide with tetra-O-benzoyl-α-D-galactopyranosyl bromide. The structure of the reaction products was confirmed by 1H and 13C NMR spectra and by chemical analysis. The ...

  2. Antioxidant, anti-alpha-glucosidase and pancreatic beta-cell protective effects of methanolic extract of Ensete superbum Cheesm seeds

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    Solomon Habtemariam

    2017-02-01

    Conclusions: The reputed antidiabetic therapeutic uses of the seeds extract of E. superbum may be justified on the basis of inhibition of carbohydrate enzymes, antioxidant effects and pancreatic β-cell protection.

  3. Antioxidant rich grape pomace extract suppresses postprandial hyperglycemia in diabetic mice by specifically inhibiting alpha-glucosidase

    Directory of Open Access Journals (Sweden)

    Hogan Shelly

    2010-08-01

    Full Text Available Abstract Background Postprandial hyperglycemia is an early defect of type 2 diabetes and one of primary anti-diabetic targets. Treatment of postprandial hyperglycemia can be achieved by inhibiting intestinal α-glucosidase, the key enzyme for oligosaccharide digestion and further glucose absorption. Grape pomace is winemaking byproduct rich in bioactive food compounds such as phenolic antioxidants. This study evaluated the anti-diabetic potential of two specific grape pomace extracts by determining their antioxidant and anti-postprandial hyperglycemic activities in vitro and in vivo. Methods The extracts of red wine grape pomace (Cabernet Franc and white wine grape pomace (Chardonnay were prepared in 80% ethanol. An extract of red apple pomace was included as a comparison. The radical scavenging activities and phenolic profiles of the pomace extracts were determined through the measurement of oxygen radical absorbance capacity, DPPH radical scavenging activity, total phenolic content and flavonoids. The inhibitory effects of the pomace extracts on yeast and rat intestinal α-glucosidases were determined. Male 6-week old C57BLKS/6NCr mice were treated with streptozocin to induce diabetes. The diabetic mice were then treated with vehicle or the grape pomace extract to determine whether the oral intake of the extract can suppress postprandial hyperglycemia through the inhibition of intestinal α-glucosidases. Results The red grape pomace extract contained significantly higher amounts of flavonoids and phenolic compounds and exerted stronger oxygen radical absorbance capacity than the red apple pomace extract. Both the grape pomace extracts but not the apple pomace extract exerted significant inhibition on intestinal α-glucosidases and the inhibition appears to be specific. In the animal study, the oral intake of the grape pomace extract (400 mg/kg body weight significantly suppressed the postprandial hyperglycemia by 35% in streptozocin-induced diabetic mice following starch challenge. Conclusion This is the first report that the grape pomace extracts selectively and significantly inhibits intestinal α-glucosidase and suppresses postprandial hyperglycemia in diabetic mice. The antioxidant and anti-postprandial hyperglycemic activities demonstrated on the tested grape pomace extract therefore suggest a potential for utilizing grape pomace-derived bioactive compounds in management of diabetes.

  4. EVALUATION OF ALPHA-AMYLASE AND ALPHA-GLUCOSIDASE INHIBITORY PROPERTIES OF SELECTED SEAWEEDS FROM GULF OF MANNAR

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    Palanisamy SenthilKumar

    2012-08-01

    Full Text Available Aqueous extracts of four seaweeds collected from Gulf of Mannar coastal waters were tested for α-amylase and α-glucosidase inhibition properties. The aqueous extracts of seaweeds in the order of Gracilaria edulis, Sargassum polycystum, Ulva lactuca and Gracilaria corticata showed significant inhibitory activity against α-amylase and α-glucosidase enzymes. G. edulis was found to be a potent inhibitor of α-glucosidase with an IC50 value of 46μg/mL. The aqueous extract of S. polycystum at a concentration of 10-100 µg/ml showed maximum α-amylase inhibitory activity with an IC50 value of 60μg/mL. This study warrants further investigation on the antidiabetic activity and identifies the hyperglycemic principle to elucidate their mode of action.

  5. Anti-inflammatory, free radical scavenging and alpha-glucosidase inhibitory activities of Hamelia patens and its chemical constituents.

    Science.gov (United States)

    Jiménez-Suárez, Verónica; Nieto-Camacho, Antonio; Jiménez-Estrada, Manuel; Alvarado Sánchez, Brenda

    2016-09-01

    Context Hamelia patens Jacq. (Rubiaceae) is traditionally used to treat wounds, inflammation and diabetes. However, there is still a lack of scientific evidence to support these applications. Objective The objective of this study is to evaluate the anti-inflammatory, antioxidant and antidiabetic activities of Hamelia patens, and identify its bioactive compounds. Materials and methods Four extracts were obtained by maceration and liquid-liquid extraction: HEX, DCM-EtOAc, MeOH-EtOAc and MeOH-Aq. The anti-inflammatory effect was evaluated orally on rat paw carrageenan-induced oedema over 6 h (50, 200 and 500 mg/kg), and topically in mouse ear oedema induced by 12-tetradecanoylphorbol-13-acetate (TPA) after 4 h (0.5 and 1 mg/ear). We also evaluated myeloperoxidase levels in ear tissue, 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging ability, and in vitro α-glucosidase inhibition. The chemical compounds were separated by column chromatography and identified by spectroscopic analysis. Results We found that the oral administration of the HEX extract at 500 and 200 mg/kg significantly decreased the carrageenan-induced inflammation after 1 and 3 h, respectively. The MeOH-EtOAc extract significantly inhibited myeloperoxidase activity (83.5%), followed by the DCM-EtOAc extract (76%), β-sitosterol/stigmasterol (72.7%) and the HEX extract (55%), which significantly decreased oedema induced by TPA at both doses, giving a similar effect to indomethacin. We also found that the MeOH-EtOAc, MeOH-Aq and DCM-EtOAc extracts showed good DPPH scavenging activity (IC50 values of 18.6, 93.9 and 158.2 μg/mL, respectively). The HEX extract showed the lowest α-glucosidase inhibition (an IC50 value of 26.07 μg/mL), followed by the MeOH-EtOAc extract (an IC50 value of 30.18 μg/mL), β-sitosterol/stigmasterol (IC50 34.6 μg/mL) and compound A ((6E,10E,14E,18E)-2,6,10,14,18,23-hexamethyl-2,6,10,14,18,22-tetracosahexaene, an IC50 value of 114.6 μg/mL), which were isolated for the first time from Hamelia patens. Discussion and conclusion Hamelia patens possesses anti-inflammatory, antioxidant and α-glucosidase inhibitory activities, which support its traditional use. These effects can be attributed to the identified compounds.

  6. Inhibitory effect of leaf extract of Newbouldia laevis on the metabolic activities of alpha-glucosidase and alpha-amylase

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    Oyetunji T. Kolawole

    2013-12-01

    Full Text Available In this study, the effect of ethanol extract of the leaves of Newbouldia laevis on the activity of α-amylase and α-glucosidase was investigated. Inhibitory effect of N. laevis extract on α-glucosidase was tested in vitro using baker’s yeast α-glucosidase and rat intestinal α-glucosidase while α-amylase inhibitory effect was assayed using rat pancreatic α-amylase. α-Glucosidase inhibitory effect of the extract was also tested in vivo in diabetic and non-diabetic rats. N. laevis extract exhibited good α-glucosidase inhibitory activity in vitro with IC50 values of 2.2 µg/mL and 43.5 µg/mL for baker’s yeast and rat intestinal α-glucosidase respectively. The extract also inhibited rat pancreatic α-amylase activity with IC50 value of 58.7 µg/mL. In both diabetic and non-diabetic rats, N. laevis extract caused a significant reduction in postprandial blood glucose level after oral sucrose load. The results of this study indicate that N. laevis extract exerts its glucose-lowering effect through inhibition of α-glucosidase and α-amylase.

  7. Alpha-amylase and alpha-glucosidase inhibition is differentially modulated by fucoidan obtained from Fucus vesiculosus and Ascophyllum nodosum.

    Science.gov (United States)

    Kim, Kyung-Tae; Rioux, Laurie-Eve; Turgeon, Sylvie L

    2014-02-01

    Fucoidan is a water-soluble, negatively charged, biologically active polysaccharide found in great abundance in brown marine algae. However, the inhibition of α-amylase and α-glucosidase by fucoidan derived from two algal species (Ascophyllum nodosum and Fucus vesiculosus) harvested at different periods (accounting for seasonal and yearly variations) has never been investigated. It was found that fucoidans inhibited α-glucosidase differently, depending on the algal species from which it was extracted and the algae's season of harvest. Fucoidan extracted from A. nodosum was a more potent inhibitor of α-glucosidase, with an IC50 ranging from 0.013 to 0.047 mg/mL, than the inhibition by fucoidan extracted from F. vesiculosus (IC50=0.049 mg/mL). In contrast, fucoidan extracted from F. vesiculosus did not inhibit α-amylase activity, while fucoidan from A. nodosum decreased α-amylase activity by 7-100% at 5 mg/mL depending upon the algae harvest period. An IC50 of 0.12-4.64 mg/mL for fucoidan from A. nodosum was found for the α-amylase inhibition. The ability of fucoidan to inhibit α-amylase and α-glucosidase thus varies according to the algae species and harvest period. A. nodosum is more suitable than F. vesiculosus as a source of fucoidan to inhibit α-amylase and α-glucosidase activities. Their potential benefits towards Type 2 diabetes management should be further investigated.

  8. Effect of storage time on metabolite profile and alpha-glucosidase inhibitory activity of Cosmos caudatus leaves – GCMS based metabolomics approach

    Directory of Open Access Journals (Sweden)

    Neda Javadi

    2015-09-01

    Full Text Available Cosmos caudatus, which is a commonly consumed vegetable in Malaysia, is locally known as “Ulam Raja”. It is a local Malaysian herb traditionally used as a food and medicinal herb to treat several maladies. Its bioactive or nutritional constituents consist of a wide range of metabolites, including glucosinolates, phenolics, amino acids, organic acids, and sugars. However, many of these metabolites are not stable and easily degraded or modified during storage. In order to investigate the metabolomics changes occurring during post-harvest storage, C. caudatus samples were subjected to seven different storage times (0 hours, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, and 12 hours at room temperature. As the model experiment, the metabolites identified by gas chromatography-mass spectrometry (GC-MS were correlated with α-glucosidase inhibitory activity analyzed with multivariate data analysis (MVDA to find out the variation among samples and metabolites contributing to the activity. Orthogonal partial least squares (OPLS analysis was applied to investigate the metabolomics changes. A profound chemical alteration, both in primary and secondary metabolites, was observed. The α-tocopherol, catechin, cyclohexen-1-carboxylic acid, benzoic acid, myo-inositol, stigmasterol, and lycopene compounds were found to be the discriminating metabolites at early storage; however, sugars such as sucrose, α-d-galactopyranose, and turanose were detected, which was attributed to the discriminating metabolites for late storage. The result shows that the MVDA method is a promising technique to identify biomarker compounds relative to storage at different times.

  9. In vitro evaluation and determination of responsible fraction of coffee beans and dried sugar beet leaves for alpha-glucosidase inhibition.

    Directory of Open Access Journals (Sweden)

    K. Singh

    2013-10-01

    Full Text Available Summary. Recent studies have identified that hydrophobic phenolic phytochemicals and hydrophilic Amadori compounds have potential for type 2 diabetes management via inhibition of carbohydrate hydrolysis enzymes. Here, we determined the phenolic content, α-glucosidase inhibitory activity, and pancreatic α-amylase inhibitory activity of water extracts of roasted and unroasted coffee beans and dried sugar beet leaves. Sugar beet leaves appeared to have the lowest total phenolic content while unroasted and roasted coffee beans had similar phenolic contents (1.49 and 1.40 mg/mL GAE DW respectively. All tested samples resulted to a dose-dependent α-glucosidase inhibitory activity. Sugar beet leaves had significant inhibitory activity (78% at the highest dose and after C18 extraction this activity appeared to be both hydrophilic and hydrophobic compound dependent.  Roasted coffee beans had significantly higher α-glucosidase inhibitory activity when compared to green coffee beans at all tested doses. Roasted coffee beans were subjected to C18 extraction and the α-glucosidase inhibitory activity was evaluated and determined to be solely hydrophobic compound dependent. When the α-amylase inhibitory activity was evaluated, no inhibition was observed with all tested samples. Our findings indicate that the observed bioactivities in coffee beans is hydrophobic compound dependent, while in sugar beet leaves the observed effect is possibly due to the synergistic effect of both hydrophobic and hydrophilic fractions. This is the first report on the carbohydrate hydrolyzing enzyme inhibition of roasted coffee beans and sugar beet leaves.Industrial Relevance. Sugar beets are widely cultivated in Europe and Northern Asia for the production of table sugar. After the harvesting of sugar beets large quantities of sugar beet leaves remain on the field and are either left to become fertilizer or appropriately disposed. Identification of appropriate strategies to utilize this by-product of sugar production, could possibly result to a sustainable utilization of this by-product. Taking into consideration that sugar is one of the reasons for the increase in the incidence of type 2 diabetes, possibly utilization of sugar-production by-products for the management of type 2 diabetes will be of extreme importance. Coffee is an important commodity and popular beverage. Over 2.25 billion cups of coffee are consumed in the world every day. Taking the above into consideration, identification of potential glucose lowering compounds in coffee can lead to the development of coffee beverages designed to prevent type 2 diabetes.Keywords: sugar beet leaves; green coffee beans; roasted coffee beans; type 2 diabetes; α-glucosidase inhibition; phenolic phytochemicals; Amadori compounds

  10. Preliminary phytochemical screening and alpha-glucosidase inhibitory activity of Philippine taro (Colocasia esculenta (L.) Schott var. PSB-VG #9)

    Science.gov (United States)

    Lebosada, Richemae Grace R.; Librando, Ivy L.

    2017-01-01

    The study was conducted to determine the anti-hyperglycemic property in terms of α-glucosidase inhibitory activity of the various parts (corm, leaf and petiole) of Colocasia esculenta (L.) Schott var. PSB-VG #9. Each of the plant parts were extracted with 95% ethanol and concentrated using a rotary evaporator at 40 °C. The crude extracts were screened for the presence of alkaloids, flavonoids, glycosides and saponins using Thin Layer Chromatography. The α-glucosidase inhibitory activity of the crude extracts (50 mg/L) were assayed spectrophotometrically using a microplate reader. The results of the phytochemical screening revealed the presence of alkaloids, flavonoids, and saponins in the leaf part while flavonoids and saponins were detected in the petiole and only saponins were present in the corm. The assay showed that the percentage α-glucosidase inhibition of the 50 mg/L ethanolic crude extract of the corm, leaves and petiole of C. esculenta are 68.03, 71.64 and 71.39%, respectively. Statistical analysis shows significant differences in the α-glucosidase inhibition among the various plant parts. It can be concluded that the ethanolic crude extracts of the different parts of C. esculenta (L.) Schott var. PSB-VG #9 exhibited inhibitory activity against α-glucosidase and the presence of phytochemicals like alkaloids, flavonoids and saponins may have contributed greatly to the inhibitory activity of the plant extract and can be further subjected for isolation of the therapeutically active compounds with antidiabetes potency.

  11. 芹菜素对鼠肠α-葡萄糖苷酶的抑制活性研究%Apigenin inhibition on alpha glucosidase in mice intestinal

    Institute of Scientific and Technical Information of China (English)

    刘春颖; 蒋淼; 屠洁; 刘冠卉

    2013-01-01

    目的:明确芹菜素对鼠肠α-葡萄糖苷酶的抑制活性.方法:建立体外α-葡萄糖苷酶抑制模型,测定芹菜素对α-葡萄糖苷酶、麦芽糖酶和蔗糖酶的抑制活性.进一步利用Lineweaver-Burk作图法,测定芹菜素对麦芽糖酶和蔗糖酶的抑制类型以及抑制常数.采用离体鼠肠进行体外模拟实验,考察芹菜素对麦芽糖酶和蔗糖酶抑制作用的稳定性.实验结果显示,芹菜素对鼠肠α-葡萄糖苷酶具有较好的抑制活性,半抑制浓度为14.475 μg/mL.芹菜素对鼠肠麦芽糖酶和蔗糖酶的半抑制浓度分别为165.35μg/mL和149.57 μg/mL,抑制类型均为竞争性抑制,抑制常数分别为32.14μg/mL和40μg/mL.在60min内,芹菜素对离体鼠肠体系中蔗糖酶和麦芽糖酶的抑制作用稳定.结论:芹菜素可竞争性抑制鼠肠麦芽糖酶和蔗糖酶的活性,可用于降低糖尿病患者的餐后血糖.%Objective:To evaluate the inhibition activities of apigenin (AP) on mice intestinal α-glucosidase.Methods:In vitro α-glucosidase inhibitory model was established to determine the inhibition activities on α-glucosidase,maltose and sucrose by AP.Furthermore,Lineweaver-Burk mapping method was adopted to determine the type of inhibition of AP.,andThe isolated mice intestine in vitro experiment was used to understand the stability of inhibition on maltose and sucrose.Results:The IC50 of AP on α-glucosidase,maltose and sucrose were 14.475μg/mL,165.35μg/mL and 149.57μg/mL,respectively.Maltose and sucrose were inhibited by AP via reversible competitiveness,and the constant of inhibition (Ki) were 32.14μg/mL and 40μg/mL,respectively.In the isolated mice intestinal system,the inhibition of AP were steady-going during 60min.Conclusions:AP can competitively inhibit mice intestinal maltase and sucrose in mice intestinal.,tTherefore,it Can bemay be used to reduce the postprandial blood glucose in diabetes patients with diabetes mellitus.

  12. The pharmacological chaperone AT2220 increases the specific activity and lysosomal delivery of mutant acid alpha-glucosidase, and promotes glycogen reduction in a transgenic mouse model of Pompe disease.

    Science.gov (United States)

    Khanna, Richie; Powe, Allan C; Lun, Yi; Soska, Rebecca; Feng, Jessie; Dhulipala, Rohini; Frascella, Michelle; Garcia, Anadina; Pellegrino, Lee J; Xu, Su; Brignol, Nastry; Toth, Matthew J; Do, Hung V; Lockhart, David J; Wustman, Brandon A; Valenzano, Kenneth J

    2014-01-01

    Pompe disease is an inherited lysosomal storage disorder that results from a deficiency in acid α-glucosidase (GAA) activity due to mutations in the GAA gene. Pompe disease is characterized by accumulation of lysosomal glycogen primarily in heart and skeletal muscles, which leads to progressive muscle weakness. We have shown previously that the small molecule pharmacological chaperone AT2220 (1-deoxynojirimycin hydrochloride, duvoglustat hydrochloride) binds and stabilizes wild-type as well as multiple mutant forms of GAA, and can lead to higher cellular levels of GAA. In this study, we examined the effect of AT2220 on mutant GAA, in vitro and in vivo, with a primary focus on the endoplasmic reticulum (ER)-retained P545L mutant form of human GAA (P545L GAA). AT2220 increased the specific activity of P545L GAA toward both natural (glycogen) and artificial substrates in vitro. Incubation with AT2220 also increased the ER export, lysosomal delivery, proteolytic processing, and stability of P545L GAA. In a new transgenic mouse model of Pompe disease that expresses human P545L on a Gaa knockout background (Tg/KO) and is characterized by reduced GAA activity and elevated glycogen levels in disease-relevant tissues, daily oral administration of AT2220 for 4 weeks resulted in significant and dose-dependent increases in mature lysosomal GAA isoforms and GAA activity in heart and skeletal muscles. Importantly, oral administration of AT2220 also resulted in significant glycogen reduction in disease-relevant tissues. Compared to daily administration, less-frequent AT2220 administration, including repeated cycles of 4 or 5 days with AT2220 followed by 3 or 2 days without drug, respectively, resulted in even greater glycogen reductions. Collectively, these data indicate that AT2220 increases the specific activity, trafficking, and lysosomal stability of P545L GAA, leads to increased levels of mature GAA in lysosomes, and promotes glycogen reduction in situ. As such, AT2220 may warrant further evaluation as a treatment for Pompe disease.

  13. 不同干酪乳杆菌菌株对α-葡萄糖苷酶抑制作用的比较%Comparison of Alpha -glucosidase Inhibitory Effect of Different Lactobacillus casei Strains

    Institute of Scientific and Technical Information of China (English)

    范文娅; 吴正钧; 季红; 郭本恒

    2012-01-01

    The α-glucosidase inhibitory activity of whey from fermented skim milk by 11 strains with milk curding ability which were selected from 14 Lactobacillus casei strains, assayed in vitro enrolling maltase ( EC 3.2.1.20) as the targeting enzyme. It was showed that the c~-glucosidase inhibitory activity of Lb. casei was strain specific, with a significant increase in inhibitory intensity with the fermentation elongated for those strains demonstrated α-glucosidase inhibitory activity. Among the tested Lb. casei strains, Lb. casei LC2W (CGMCC0828) demonstrated the strongest α- glucosidase inhibitory activity, with an inhibitory percentage of 37.36% for whey derived of 10% (w/w) skim milk fermented at 37C for 96 hours. These results suggested Lb. casei LC2W has the potentials of antihyperglycemia as well as anti-diabetes.%从14株干酪乳杆菌中筛选出11株具有凝乳作用的菌株,采用以麦芽糖酶为靶向酶的α-葡萄糖苷酶抑制剂体外筛选模型,分析了不同干酪乳杆菌发酵脱脂乳上清对α-葡萄糖苷酶的抑制作用。结果表明,不同菌株发酵脱脂乳上清对麦芽糖酶的抑制活性具有明显的菌株特异性,且随着发酵时间延长,α-葡萄糖苷酶抑制活性呈现上升趋势。其中,干酪乳杆菌LC2W在37℃发酵10%(w/w)脱脂乳96 h后所得的上清液对麦芽糖酶抑制活性最高,达37.36%,明显高于其它被测试的干酪乳杆菌菌株,表明该菌株具有潜在的抗高血糖作用。

  14. Synthèses enzymatiques de néoglucoconjugués catalysées par l'alpha-glucosidase purifiée de la blatte Periplaneta americana (Linnaeus

    Directory of Open Access Journals (Sweden)

    Kamenan A.

    2005-01-01

    Full Text Available Enzymatic synthesis of neoglucoconjugates by purified α-glucosidase from cockroach Periplaneta americana (Linnaeus. Cockroach Periplaneta americana (Linnaeus contains in his digestive tract an acid (pH 5,0 and mesophile (50°C α-glucosidase. This enzyme, purified to homogeneity, hydrolyses highly maltose, sucrose and p-nitrophenyl-α-Dglucopyranoside. The ability of α-glucosidase from cockroach purified to homogeneity to catalyse transglucosylation reactions was tested using maltose and saccharose as glucosyl donors and 2-phenylethanol and phenol as acceptors. The experimental conditions were optimized in relation to the time course of the reaction, pH and concentrations of glucosyl donors and acceptors. The yields in transglucosylation reactions at 37 °C were very high and could attain 67% and 48% with 2-phenylethanol and phenol respectively as glucosyl acceptors. This α-glucosidase hydrolyzed the products formed. It seems that the products formed were the phenylethyl-α-D-glucoside and phenyl-α-D-glucoside. These results suggest that α- glucosidase from cockroach is an exoglucosidase which catalyse the splitting of the α-glucosyl residue from the non reducing terminal of the substrate to liberate α-glucose. This comportment indicates that this enzyme operated by a mechanism involving the retention of the anomeric configuration. On the basis of this work, α-glucosidase from P. americana appears to be a valuable tool for the preparation of α-neoglucoconjugates.

  15. Alpha-glucosidase Inhibitors from Fermented Soybean Milk and Steamed Soybean by Lactobacillus plantarum ST-Ⅲ%植物乳酸菌ST-Ⅲ产α-糖苷酶抑制剂作用的研究

    Institute of Scientific and Technical Information of China (English)

    韩瑨; 季红; 周方方; 吴正钧

    2014-01-01

    以改良的体外模型为测定手段,评价了植物乳杆菌ST-Ⅲ(Lactobacillus plantarum ST-Ⅲ)以蒸煮大豆为培养基料时,其接种量、发酵温度对发酵产物抑制麦芽糖酶的影响,同时,通过对植物乳杆菌ST-Ⅲ在最优条件下发酵大豆豆浆和蒸煮大豆的产物对麦芽糖酶的抑制效果及其相应粗提物对麦芽糖酶的半数抑制浓度IC50的比较,评价了植物乳杆菌ST-Ⅲ在不同形式的大豆基料中产-葡萄糖苷酶抑制剂的能力,结果表明:植物乳杆菌ST-Ⅲ发酵蒸煮大豆产生麦芽糖酶抑制物的最适条件分别为接种量2%(v/w)、发酵温度37℃。在优化条件下,植物乳杆菌ST-Ⅲ发酵大豆豆浆10 h的产物对麦芽糖酶的抑制效果可达80%左右,而发酵蒸煮大豆时达到相同抑制效果需发酵10 d以上。发酵两种不同形式大豆基料制得的粗提物对麦芽糖酶的半数抑制浓度IC50分别为2.989 mg/mL和2.597 mg/mL。因此,液体状态的大豆豆浆是植物乳杆菌ST-Ⅲ发酵产-葡萄糖苷酶抑制剂的优选基料形式。%The optimum condition for Lactobacillus plantarum ST-Ⅲto biosynthesize maltase (EC 3.2.1.20) inhibitors in fermented steamed soybean was assayed via a modified model in vitro , and the optimized condition was found to be a combination of inoculation ratio of 2%(v/w), fermentation temperature of 37℃, respectively. Under the optimized conditions, inhibitory effect on maltase of soybean milk fermented 10 h reached 80%, and the time for the fermented steamed soybean reaching the same inhibitory effect on maltase is at least 10 d. The IC 50 of crude extract prepared from soybean milk and steamed soybean fermented by Lactobacillus plantarum ST-Ⅲwas 2.989 mg/mL and 2.597 mg/mL. Therefore, soybean milk was the more suitable form for Lactobacillus plantarum ST-Ⅲto produceα-glucosidase inhibitors.

  16. Serviceberry [Amerlanchier alnifolia (Nutt.) Nutt. ex. M. Roem(Rosaceae)] leaf exhibits mammalian alpha glucosidase activity and suppresses postprandial glycemic response in a mouse model of diet induced obesity/hyperglycemia

    Science.gov (United States)

    Several plant-based remedies offer cost-effective management of diabetes, but few plant species adapted to North America have been validated for their antidiabetic properties. One such species is serviceberry (Amelanchier alnifolia), found in Browning, MT, which has been traditionally used by the Am...

  17. Studies on Enzymatic Properties of Alpha-glucosidase in Aspergillus niger CICIM F0410%黑曲霉CICIM F0410中α-葡萄糖苷酶的酶学性质研究

    Institute of Scientific and Technical Information of China (English)

    周敏; 王正祥

    2009-01-01

    研究了黑曲霉(Aspergillus niger)CICIM F0410发酵粗酶液中α-葡萄糖苷酶的酶学性质,发现该酶最适反应温度为55℃,最适反应pH值5.0,在65℃以下保持酶活力相对稳定,且能在pH值(4.0~6.5)范围内保持相对较高的酶活力.以该菌株的染色体DNA为模板,PCR扩增出大小为3.1 kb的片段,经序列比对发现与NCBI上已公布的黑曲霉(Aspergillus niger)CBS513.88(登录号XP_001402053)序列有2个碱基的区别:2272(A→G),3098(T→G),导致2个氨基酸残基的变化:687(Asn→ser)、983(Ser→Arg).

  18. SwissProt search result: AK102309 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102309 J033090B18 (P10253) Lysosomal alpha-glucosidase precursor (EC 3.2.1.20) (Acid maltase) (Aglucosidas...e alfa) [Contains: 76 kDa lysosomal alpha-glucosidase; 70 kDa lysosomal alpha-glucosidase] LYAG_HUMAN 1e-145 ...

  19. SwissProt search result: AK066051 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK066051 J013051B02 (P10253) Lysosomal alpha-glucosidase precursor (EC 3.2.1.20) (Acid maltase) (Aglucosidas...e alfa) [Contains: 76 kDa lysosomal alpha-glucosidase; 70 kDa lysosomal alpha-glucosidase] LYAG_HUMAN 1e-89 ...

  20. Efecto de enzimas proteolíticas y de la abstinencia sexual en la cuantificación bioquímica de alfa-glucosidasa en plasma seminal humano

    OpenAIRE

    2001-01-01

    Background: alpha-glucosidase is found in human seminal plasma as an acid form, located in accessory glands, and as a neutral form secreted almost exclusively by the epididymis. Quantification of alpha-glucosidase activity is a marker of the secretory function of the epididymis and indemnity of the sperm transport pathway Aim: To obtain reference values for alpha-glucosidase in normal samples of seminal plasma, to evaluate its behavior in serial samples and to determine the effect of proteoly...

  1. Sequence Classification: 407438 [

    Lifescience Database Archive (English)

    Full Text Available TMB Non-TMH TMB TMB TMB Non-TMB >gi|15828934|ref|NP_326294.1| GLUCAN 1,6-ALPHA-GLUCOSIDASE (DEXTRAN GLUCOSID...ASE) (EXO-1,6-ALPHA-GLUCOSIDASE) (GLUCODEXTRANASE) || http://www.ncbi.nlm.nih.gov/protein/15828934 ...

  2. NCBI nr-aa BLAST: CBRC-CINT-01-0028 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CINT-01-0028 gb|EAW83988.1| maltase-glucoamylase (alpha-glucosidase), isoform ...CRA_a [Homo sapiens] gb|EAW83989.1| maltase-glucoamylase (alpha-glucosidase), isoform CRA_a [Homo sapiens] EAW83988.1 4.3 29% ...

  3. Effect of enzyme therapy in juvenile patients with Pompe disease: a three-year open-label study.

    NARCIS (Netherlands)

    Capelle, C.I. van; Beek, N.A. van der; Hagemans, M.L.; Arts, W.F.M.; Hop, W.C.J.; Lee, P.; Jaeken, J.; Frohn-Mulder, I.M.; Merkus, P.J.F.M.; Corzo, D.; Puga, A.C.; Reuser, A.J.J.; Ploeg, A.T. van der

    2010-01-01

    Pompe disease is a rare neuromuscular disorder caused by deficiency of acid alpha-glucosidase. Treatment with recombinant human alpha-glucosidase recently received marketing approval based on prolonged survival of affected infants. The current open-label study was performed to evaluate the response

  4. NCBI nr-aa BLAST: CBRC-DNOV-01-1573 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-1573 ref|ZP_01446186.1| probable alpha-glucosidase protein [Roseovariu...s sp. HTCC2601] gb|EAU43606.1| probable alpha-glucosidase protein [Roseovarius sp. HTCC2601] ZP_01446186.1 0.59 38% ...

  5. Sequence Classification: 398894 [

    Lifescience Database Archive (English)

    Full Text Available BLE ALPHA-GLUCOSIDASE AGLA (MALTASE) (GLUCOINVERTASE) (GLUCOSIDOSUCRASE) (MALTASE-GLUCOAMYLASE) (LYSOSOMAL A...LPHA-GLUCOSIDASE) (ACID MALTASE) || http://www.ncbi.nlm.nih.gov/protein/15609608 ...

  6. Sequence Classification: 389141 [

    Lifescience Database Archive (English)

    Full Text Available BLE ALPHA-GLUCOSIDASE AGLA (MALTASE) (GLUCOINVERTASE) (GLUCOSIDOSUCRASE) (MALTASE-GLUCOAMYLASE) (LYSOSOMAL A...LPHA-GLUCOSIDASE) (ACID MALTASE) || http://www.ncbi.nlm.nih.gov/protein/31793652 ...

  7. Type 2 diabetes

    Science.gov (United States)

    ... medicines (GLP-1 analogs) Meglitinides SGL T2 inhibitors Sulfonylureas Thiazolidinediones You may need to take insulin if ... rule Glucose in blood Alpha-glucosidase inhibitors Biguanides Sulfonylureas drug Thiazolidinediones Food and insulin release Monitoring blood ...

  8. Antidiabetic potential of Conocarpus lancifolius

    Directory of Open Access Journals (Sweden)

    Malik Saadullah

    2014-06-01

    Full Text Available The antidiabetic activity of Conocarpus lancifolius extract was investigated in vitro, as alpha glucosidase inhibition and in vivo as alloxan induced diabetic rabbits with other biochemical parameters (LDL, HDL, SGPT, SGOT, cretinine, urea and triglyceride. Alpha-glucosidase inhibition activity was performed by using acorbose as standred. Methanolic extract show alpha-glucosidase inhibition activity. The dose of 200 mg/kg body weight significantly (p<0.05 decreases the blood glucose level, plasma total cholesterol, triglycerides and LDL in treated rabbits as compared to diabetic rabbits. This dose significantly increased the level of HDL in treated group. The activity of SGOT and SGPT also significantly (p<0.05 decreased in treated diabetic rabbits. Phytochemical studies show the presence of glycosides, tannins, saponins and terpenoids. The antidiabetic potential is may be due to its saponin contents.

  9. A case of childhood Pompe disease demonstrating phenotypic variability of p.Asp645Asn.

    Science.gov (United States)

    Kroos, Marian A; Kirschner, Janbernd; Gellerich, Frank N; Hermans, Monique M P; Van Der Ploeg, Ans T; Reuser, Arnold J J; Korinthenberg, Rudolf

    2004-06-01

    A six-year-old child presented at 8 months of age with proximal muscle weakness and mild cardiac hypertrophy. Some alpha-glucosidase activity was detected in muscle but not in fibroblasts. As none of the two pathogenic mutations, [c.1933G>A]+[c.2702T>A] (Asp645Asn/Leu901Gln), led to detectable alpha-glucosidase activity upon expression in COS cells, the phenotype of the patient remained unexplained. A functionally comparable set of mutations, Asp645Asn/insGnt2243, was reported previously to cause classic infantile Pompe disease [Biochem Biophys Res Commun 244 (1998) 921]. We conclude that secondary genetic or environmental factors can be decisive for the phenotypic outcome of classic infantile versus childhood Pompe disease, when the acid alpha-glucosidase activity is extremely low.

  10. Dietary phenolic compounds selectively inhibit the individual subunits of maltase-glucoamylase and sucrase-isomaltase with the potential of modulating glucose release

    Science.gov (United States)

    In this study, it was hypothesized that dietary phenolic compounds selectively inhibit the individual C- and N-terminal (Ct, Nt) subunits of the two small intestinal alpha-glucosidases, maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI), for a modulated glycemic carbohydrate digestion. The inhi...

  11. InterProScan Result: BP124820 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BP124820 BP124820_3_ORF2 35CD3B2AD11B04C6 PANTHER PTHR22762:SF7 NEUTRAL ALPHA-GLUCOSIDA...SE AB PRECURSOR (GLUCOSIDASE II ALPHA SUBUNIT) (ALPHA GLUCOSIDASE 2) NA ? IPR000322 unintegrated ...

  12. InterProScan Result: BP124820 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BP124820 BP124820_5_ORF1 DC1B7DDE14643E5E PANTHER PTHR22762:SF7 NEUTRAL ALPHA-GLUCOSIDA...SE AB PRECURSOR (GLUCOSIDASE II ALPHA SUBUNIT) (ALPHA GLUCOSIDASE 2) NA ? IPR000322 unintegrated ...

  13. InterProScan Result: FS830859 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS830859 FS830859_4_ORF1 D905399DCEA8EB3E PANTHER PTHR22762:SF7 NEUTRAL ALPHA-GLUCOSIDA...SE AB PRECURSOR (GLUCOSIDASE II ALPHA SUBUNIT) (ALPHA GLUCOSIDASE 2) NA ? IPR000322 unintegrated Biological Process: carbohydrate metabolic process (GO:0005975) ...

  14. InterProScan Result: BP124820 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BP124820 BP124820_2_ORF1 DFD3D5A0B4996BA2 PANTHER PTHR22762:SF7 NEUTRAL ALPHA-GLUCOSIDA...SE AB PRECURSOR (GLUCOSIDASE II ALPHA SUBUNIT) (ALPHA GLUCOSIDASE 2) 3.9e-72 T IPR000322 unintegrated Biological Process: carbohydrate metabolic process (GO:0005975) ...

  15. InterProScan Result: FS830859 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS830859 FS830859_4_ORF1 D905399DCEA8EB3E PANTHER PTHR22762:SF7 NEUTRAL ALPHA-GLUCOSIDA...SE AB PRECURSOR (GLUCOSIDASE II ALPHA SUBUNIT) (ALPHA GLUCOSIDASE 2) 9.9e-21 T IPR000322 unintegrated Biological Process: carbohydrate metabolic process (GO:0005975) ...

  16. InterProScan Result: CK495917 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available CK495917 CK495917_5_ORF1 7F944B343AFB57FA PANTHER PTHR22762:SF7 NEUTRAL ALPHA-GLUCOSIDA...SE AB PRECURSOR (GLUCOSIDASE II ALPHA SUBUNIT) (ALPHA GLUCOSIDASE 2) 6.1e-18 T IPR000322 unintegrated Biological Process: carbohydrate metabolic process (GO:0005975) ...

  17. InterProScan Result: BP124820 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available BP124820 BP124820_6_ORF2 8E9FAE9903327F4B PANTHER PTHR22762:SF7 NEUTRAL ALPHA-GLUCOSIDA...SE AB PRECURSOR (GLUCOSIDASE II ALPHA SUBUNIT) (ALPHA GLUCOSIDASE 2) NA ? IPR000322 unintegrated ...

  18. Pompe disease : Current state of treatment modalities and animal models

    NARCIS (Netherlands)

    Geel, T. M.; McLaughlin, P. M. J.; de Leij, L. F. M. H.; Ruiters, M. H. J.; Niezen-Koning, K. E.

    2007-01-01

    Pompe disease is a rare autosomal recessive lysosomal storage disease caused by deficiency of acid-alpha-glucosidase (GAA). This deficiency results in glycogen accumulation in the lysosomes, leading to lysosomal swelling, cellular damage and organ dysfunction. In early-onset patients (the classical

  19. Effect of miglitol (BAY m-1099) on fasting blood glucose in type 2 diabetes mellitus

    NARCIS (Netherlands)

    Sels, J P; Kingma, P J; Wolffenbuttel, B H; Menheere, P P; Branolte, J H; Nieuwenhuijzen Kruseman, A C

    1994-01-01

    BACKGROUND: In type 2 diabetes mellitus, fasting blood glucose values are increased due to increased glycogenolysis and gluconeogenesis. As miglitol (BAY m-1099), an absorbable alpha-glucosidase inhibitor, can inhibit glycogenolysis, we investigated whether 200 mg miglitol ingested at bedtime could

  20. Disposition kinetics and bioavailability of the glucosidase inhibitor N-benzyl-1-deoxynojirimycin after various routes of administration in mice

    NARCIS (Netherlands)

    Faber, ED; Delbressine, LPC; vandeVorstenbosch, CG; vandenBroek, LAGM; Meijer, DKF; Stok, B.P.

    1997-01-01

    Pharmacokinetics, biodistribution, and excretion of the alpha-glucosidase inhibitor and antiviral compound N-benzyl-1-deoxynojirimycin (BndNM) were studied in mice, after intravenous, subcutaneous, and oral administration of a single radiolabeled dose, No metabolites were detected in plasma, urine,

  1. Inhibition of recombinant human maltase glucoamylase by salacinol and derivatives.

    Science.gov (United States)

    Rossi, Elena J; Sim, Lyann; Kuntz, Douglas A; Hahn, Dagmar; Johnston, Blair D; Ghavami, Ahmad; Szczepina, Monica G; Kumar, Nag S; Sterchi, Erwin E; Nichols, Buford L; Pinto, B M; Rose, David R

    2006-06-01

    Inhibitors targeting pancreatic alpha-amylase and intestinal alpha-glucosidases delay glucose production following digestion and are currently used in the treatment of Type II diabetes. Maltase-glucoamylase (MGA), a family 31 glycoside hydrolase, is an alpha-glucosidase anchored in the membrane of small intestinal epithelial cells responsible for the final step of mammalian starch digestion leading to the release of glucose. This paper reports the production and purification of active human recombinant MGA amino terminal catalytic domain (MGAnt) from two different eukaryotic cell culture systems. MGAnt overexpressed in Drosophila cells was of quality and quantity suitable for kinetic and inhibition studies as well as future structural studies. Inhibition of MGAnt was tested with a group of prospective alpha-glucosidase inhibitors modeled after salacinol, a naturally occurring alpha-glucosidase inhibitor, and acarbose, a currently prescribed antidiabetic agent. Four synthetic inhibitors that bind and inhibit MGAnt activity better than acarbose, and at comparable levels to salacinol, were found. The inhibitors are derivatives of salacinol that contain either a selenium atom in place of sulfur in the five-membered ring, or a longer polyhydroxylated, sulfated chain than salacinol. Six-membered ring derivatives of salacinol and compounds modeled after miglitol were much less effective as MGAnt inhibitors. These results provide information on the inhibitory profile of MGAnt that will guide the development of new compounds having antidiabetic activity.

  2. Lentiviral gene therapy of murine hematopoietic stem cells ameliorates the Pompe disease phenotype.

    Science.gov (United States)

    van Til, Niek P; Stok, Merel; Aerts Kaya, Fatima S F; de Waard, Monique C; Farahbakhshian, Elnaz; Visser, Trudi P; Kroos, Marian A; Jacobs, Edwin H; Willart, Monique A; van der Wegen, Pascal; Scholte, Bob J; Lambrecht, Bart N; Duncker, Dirk J; van der Ploeg, Ans T; Reuser, Arnold J J; Verstegen, Monique M; Wagemaker, Gerard

    2010-07-01

    Pompe disease (acid alpha-glucosidase deficiency) is a lysosomal glycogen storage disorder characterized in its most severe early-onset form by rapidly progressive muscle weakness and mortality within the first year of life due to cardiac and respiratory failure. Enzyme replacement therapy prolongs the life of affected infants and supports the condition of older children and adults but entails lifelong treatment and can be counteracted by immune responses to the recombinant enzyme. We have explored the potential of lentiviral vector-mediated expression of human acid alpha-glucosidase in hematopoietic stem cells (HSCs) in a Pompe mouse model. After mild conditioning, transplantation of genetically engineered HSCs resulted in stable chimerism of approximately 35% hematopoietic cells that overexpress acid alpha-glucosidase and in major clearance of glycogen in heart, diaphragm, spleen, and liver. Cardiac remodeling was reversed, and respiratory function, skeletal muscle strength, and motor performance improved. Overexpression of acid alpha-glucosidase did not affect overall hematopoietic cell function and led to immune tolerance as shown by challenge with the human recombinant protein. On the basis of the prominent and sustained therapeutic efficacy without adverse events in mice we conclude that ex vivo HSC gene therapy is a treatment option worthwhile to pursue.

  3. Enzyme replacement therapy in late-onset Pompe's disease: a three-year follow-up.

    Science.gov (United States)

    Winkel, Léon P F; Van den Hout, Johanna M P; Kamphoven, Joep H J; Disseldorp, Janus A M; Remmerswaal, Maaike; Arts, Willem F M; Loonen, M Christa B; Vulto, Arnold G; Van Doorn, Pieter A; De Jong, Gerard; Hop, Wim; Smit, G Peter A; Shapira, Stuart K; Boer, Marijke A; van Diggelen, Otto P; Reuser, Arnold J J; Van der Ploeg, Ans T

    2004-04-01

    Pompe's disease is an autosomal recessive myopathy. The characteristic lysosomal storage of glycogen is caused by acid alpha-glucosidase deficiency. Patients with late-onset Pompe's disease present with progressive muscle weakness also affecting pulmonary function. In search of a treatment, we investigated the feasibility of enzyme replacement therapy with recombinant human alpha-glucosidase from rabbit milk. Three patients (aged 11, 16, and 32 years) were enrolled in the study. They were all wheelchair-bound and two of them were ventilator dependent with a history of deteriorating pulmonary function. After 3 years of treatment with weekly infusions of alpha-glucosidase, the patients had stabilized pulmonary function and reported less fatigue. The youngest and least affected patient showed an impressive improvement of skeletal muscle strength and function. After 72 weeks of treatment, he could walk without support and finally abandoned his wheelchair. Our findings demonstrate that recombinant human alpha-glucosidase from rabbit milk has a therapeutic effect in late-onset Pompe's disease. There is good reason to continue the development of enzyme replacement therapy for Pompe's disease and to explore further the production of human therapeutic proteins in the milk of mammals.

  4. AcEST: DK956317 [AcEST

    Lifescience Database Archive (English)

    Full Text Available . 40 0.062 tr|A2QHE1|A2QHE1_ASPNC Glucan 1,4-alpha-glucosidase glaA-Aspergi... 40 0.062 tr|Q52516|Q52516_PSESP Maltopentaos...QACGTSTATVTDTW 639 >tr|Q52516|Q52516_PSESP Maltopentaose forming amylase OS=Pseudomonas sp. PE=3 SV=1 Length

  5. Low-dose acarbose does not delay digestion of starch but reduces its bioavailability

    NARCIS (Netherlands)

    Wachters-Hagedoorn, R. E.; Priebe, M. G.; Heimweg, J. A. J.; Heiner, A. M.; Elzinga, H.; Stellaard, F.; Vonk, R. J.

    2007-01-01

    Aims Slowly digestible starch is associated with beneficial health effects. The glucose-lowering drug acarbose has the potential to retard starch digestion since it inhibits alpha-amylase and alpha-glucosidases. We tested the hypothesis that a low dose of acarbose delays the rate of digestion of rap

  6. In vitro inhibition activity of polyphenol-rich extracts from Syzygium aromaticum (L.) Merr. & Perry (Clove) buds against carbohydrate hydrolyzing enzymes linked to type 2 diabetes and Fe2+-induced lipid peroxidation in rat pancreas

    Institute of Scientific and Technical Information of China (English)

    Stephen Adeniyi Adefegha; Ganiyu Oboh

    2012-01-01

    To investigate and compare the inhibitory properties of free and bound phenolic extracts of clove bud against carbohydrate hydrolyzing enzymes (alpha-amylase & alpha-glucosidase) and Fe2+-induced lipid peroxidation in rat pancreas in vitro. Methods: The free phenolics were extracted with 80% (v/v) acetone, while bound phenolics were extracted from the alkaline and acid hydrolyzed residue with ethyl acetate. Then, the interaction of the extracts with alpha-amylase and alpha-glucosidase was subsequently assessed. Thereafter, the total phenolic contents and antioxidant activities of the extracts were determined. Results: The result revealed that both extracts inhibited alpha-amylase and alpha-glucosidase in a dose-dependent manner. However, the alpha-glucosidase inhibitory activity of the extracts were significantly (P<0.05) higher than their alpha-amylase inhibitory activity. The free phenolics (31.67 mg/g) and flavonoid (17.28 mg/g) contents were significantly (P<0.05) higher than bound phenolic (23.52 mg/g) and flavonoid (13.70 mg/g) contents. Both extracts also exhibited high antioxidant activities as typified by their high reducing power, 1,1 diphenyl-2- picrylhydrazyl (DPPH) and 2, 2-azinobis-3-ethylbenzo-thiazoline-6-sulfonate (ABTS) radical scavenging abilities, as well as inhibition of Fe2+-induced lipid peroxidation in rat pancreas in vitro. Conclusions: This study provides a biochemical rationale by which clove elicits therapeutic effect on type 2 diabetes.

  7. Gclust Server: 96223 [Gclust Server

    Lifescience Database Archive (English)

    Full Text Available alpha-glucosidase (sucrase-maltase-isomaltase) 1 1.00e-28 0.0 0.0 0.0 0.0 3.23 0.0 Show 96223 Cluster ID 962...ha-glucosidase (sucrase-maltase-isomaltase) Number of Sequences 1 Homologs 1 Clustering threshold 1.00e-28 P

  8. [Evaluation of the association between the use of oral anti-hyperglycemic agents and hypoglycemia in Japan by data mining of the Japanese Adverse Drug Event Report (JADER) database].

    Science.gov (United States)

    Umetsu, Ryogo; Nishibata, Yuri; Abe, Junko; Suzuki, Yukiya; Hara, Hideaki; Nagasawa, Hideko; Kinosada, Yasutomi; Nakamura, Mitsuhiro

    2014-01-01

    Hypoglycemia due to treatment with oral anti-hyperglycemic agents (OHAs) is a major clinical problem in patients with type 2 diabetes mellitus. The aim of the present study was to evaluate the risk of hypoglycemia due to OHA use by using the Japanese Adverse Drug Event Report (JADER) database. To this end, reports of hypoglycemia events included in the JADER database between 2004 and 2012 were analyzed by calculating the reporting odds ratio (OR). The Medical Dictionary for Regulatory Activities Preferred Terms was used to identify hypoglycemia; 254392 reports were found in the JADER database, of which 13269 were excluded because the age and sex of the patient were not reported. Finally, 241123 reports were analyzed. Among OHAs, sulfonylureas showed the highest adjusted OR (adjusted OR, 10.13; 95% confidence interval, 9.08-11.26). The adjusted ORs for meglitinides, biguanide, thiazolidinedione, alpha-glucosidase inhibitors, and dipeptidyl peptidase-4 inhibitors were significantly lower than that of sulfonylureas. The adjusted OR of meglitinides (3.17; 95% confidence interval, 2.23-4.36) was significantly higher than that of alpha-glucosidase inhibitors or thiazolidinedione. We observed no difference between the adjusted ORs for biguanide, thiazolidinedione, alpha-glucosidase inhibitors, and dipeptidyl peptidase-4 inhibitors. Data mining of the JADER database was useful for analyzing OHA-associated hypoglycemia events. The results of our study suggested a low risk of hypoglycemia associated with biguanide, thiazolidinedione, alpha-glucosidase inhibitors, and dipeptidyl peptidase-4 inhibitors in clinical practice.

  9. Enzymic synthesis of alpha- and beta-D-glucosides of 1-deoxynojirimycin and their glycosidase inhibitory activities.

    Science.gov (United States)

    Asano, N; Oseki, K; Kaneko, E; Matsui, K

    1994-05-20

    1-Deoxynojirimycin (1) is a potent inhibitor of mammalian and rice alpha-glucosidase. Several glucosides of 1 were synthesized by use of the native and immobilized enzyme and their effect on various enzymes was investigated. Transglucosylation reactions using rice alpha-glucosidase, yeast alpha- and beta-glucosidases purified from Rhodotorula lactosa were performed with maltose or cellobiose as a glucose donor and N-(benzyloxycarbonyl)-1-deoxynojirimycin (2) as an acceptor. The transglucosylation reaction using native rice alpha-glucosidase afforded 3-O-alpha-D-glucopyranosyl-N-(benzyloxycarbonyl)-1-deoxynojirimycin (4), 4-O-alpha-D-glucopyranosyl-N-(benzyloxycarbonyl)-1-deoxynojirimycin (5), and 2-O-alpha-D-glucopyranosyl-N-(benzyloxycarbonyl)-1-deoxynojirimycin (3) in yields of 40, 13, and 2%, respectively, after 30 min. The transglucosylation reaction using immobilized rice alpha-glucosidase was similar to that using the native enzyme. In the system using native yeast alpha-glucosidase, 3, 5, and 4 were formed in yields of 34, 13, and 6%, respectively, after 15 h. The immobilization of yeast alpha-glucosidase caused a significant decrease in transglucosylation activity. Yeast beta-glucosidase showed a high transglucosylation activity and incubation with the reaction system afforded 2-O-beta-D-glucopyranosyl-N-(benzyloxycarbonyl)-1-deoxynojirimycin (6) and 4-O-beta-D-glucopyranosyl-N-(benzyloxycarbonyl)-1-deoxynojirimycin (7) in yields of 69 and 3%, respectively, after 3 h. The transglucosylation reaction using immobilized yeast beta-glucosidase preferentially afforded 6 in a yield of 73% after 3 h. After removal of N-benzyloxycarbonyl group from the product glucosides, their glycosidase inhibitory activities were measured. 3-O-alpha-D-Glucopyranosyl-1-deoxynojirimycin (9) retained the potent inhibition of 1 against rat intestinal sucrase activity and was more effective than 1 against rice alpha-glucosidase. 4-O-alpha-D-Glucopyranosyl-1-deoxynojirimycin (10

  10. Evaluation of Rhodiola crenulata and Rhodiola rosea for management of type II diabetes and hypertension.

    Science.gov (United States)

    Kwon, Young-In; Jang, Hae-Dong; Shetty, Kalidas

    2006-01-01

    In the current study, we investigated 2 species of the genus Rhodiola for the inhibition of alpha-amylase,alpha-glucosidase and angiotensin converting enzyme (ACE) inhibitory activity. Water extracts of Rhodiola crenulata had the highest alpha-amylase inhibitory activity (IC50,98.1 microg total phenolic /ml) followed by ethanol extract of R.crenulata (IC50, 120.9 microg total phenolic/ml) and ethanol extract of R.rosea (IC50, 173.4 microg total phenolic /ml). Ethanol R.rosea (IC50, 44.7 microg total phenolic/ml), water extract of R.rosea (IC50, 52.3 microg total phenolic/ml), water extract of R.crenulata (IC50, 60.3 microg total phenolic /ml) and ethanol extract of R.crenulata (IC50, 60.2 microg total phenolic/ml) also showed significant alpha-glucosidase inhibitory activity. The alpha-glucosidase inhibitory activity of the extracts was compared to standard tyrosol, which was significantly detected in the extracts using HPLC. Tyrosol had strong alpha-glucosidase inhibitory activity (IC50, 70.8 microg total phenolic/ml) but did not have any inhibitory effect on the alpha-amylase activity. Results suggested that alpha-glucosidase inhibitory activities of both Rhodiola extracts correlated to the phenolic content, antioxidant activity and phenolic profile of the extracts. The ability of the above Rhodiola extracts to inhibit rabbit lung angiotensin I-converting enzyme (ACE) was investigated. The ethanol extracts of R.rosea had the highest ACE inhibitory activity (38.5 %) followed by water extract of R.rosea (36.2 %) and R.crenulata (15.4 %).

  11. Determinación de la variabilidad en la condensación de la cromatina espermática y alfa-glucosidasa seminal con relación al espermiograma

    OpenAIRE

    2000-01-01

    Background: Sperm functional tests as an addition to semen analysis have been used to study the fertilization ability of spermatozoa. Besides the usual variability of the seminal analysis an individual variability in the results of functional tests has been recently found. Aim: to evaluate in a three months period, the individual variability of sperm parameters and sperm maturation using the chromatin condensation test and epidydime alpha-glucosidase (that allows to discriminate obstructive p...

  12. Pompe disease: literature review and case series.

    Science.gov (United States)

    Dasouki, Majed; Jawdat, Omar; Almadhoun, Osama; Pasnoor, Mamatha; McVey, April L; Abuzinadah, Ahmad; Herbelin, Laura; Barohn, Richard J; Dimachkie, Mazen M

    2014-08-01

    Pompe disease is a rare multi-systemic metabolic myopathy caused by autosomal recessive mutations in the acidic alpha glucosidase (GAA) gene. Significant progress had been made in the diagnosis and management of patients with Pompe disease. Here, we describe our experience with 12 patients with various forms of Pompe disease including 4 potentially pathogenic, novel GAA variants. We also review the recent the recent advances in the pathogenesis, diagnosis, and treatment of individuals with Pompe disease.

  13. Measurement of alpha-amylase activity in white wheat flour, milled malt, and microbial enzyme preparations, using the Ceralpha assay: collaborative study.

    Science.gov (United States)

    McCleary, Barry V; McNally, Marian; Monaghan, Dympna; Mugford, David C

    2002-01-01

    This study was conducted to evaluate the method performance of a rapid procedure for the measurement of alpha-amylase activity in flours and microbial enzyme preparations. Samples were milled (if necessary) to pass a 0.5 mm sieve and then extracted with a buffer/salt solution, and the extracts were clarified and diluted. Aliquots of diluted extract (containing alpha-amylase) were incubated with substrate mixture under defined conditions of pH, temperature, and time. The substrate used was nonreducing end-blocked p-nitrophenyl maltoheptaoside (BPNPG7) in the presence of excess quantities of thermostable alpha-glucosidase. The blocking group in BPNPG7 prevents hydrolysis of this substrate by exo-acting enzymes such as amyloglucosidase, alpha-glucosidase, and beta-amylase. When the substrate is cleaved by endo-acting alpha-amylase, the nitrophenyl oligosaccharide is immediately and completely hydrolyzed to p-nitrophenol and free glucose by the excess quantities of alpha-glucosidase present in the substrate mixture. The reaction is terminated, and the phenolate color developed by the addition of an alkaline solution is measured at 400 nm. Amylase activity is expressed in terms of Ceralpha units; 1 unit is defined as the amount of enzyme required to release 1 micromol p-nitrophenyl (in the presence of excess quantities of alpha-glucosidase) in 1 min at 40 degrees C. In the present study, 15 laboratories analyzed 16 samples as blind duplicates. The analyzed samples were white wheat flour, white wheat flour to which fungal alpha-amylase had been added, milled malt, and fungal and bacterial enzyme preparations. Repeatability relative standard deviations ranged from 1.4 to 14.4%, and reproducibility relative standard deviations ranged from 5.0 to 16.7%.

  14. Morphological changes in muscle tissue of patients with infantile Pompe's disease receiving enzyme replacement therapy.

    Science.gov (United States)

    Winkel, Léon P F; Kamphoven, Joep H J; van den Hout, Hannerieke J M P; Severijnen, Lies A; van Doorn, Pieter A; Reuser, Arnold J J; van der Ploeg, Ans T

    2003-06-01

    Pompe's disease (glycogen storage disease type II) is an autosomal recessive myopathy caused by lysosomal alpha-glucosidase deficiency. Enzyme replacement therapy (ERT) is currently under development for this disease. We evaluated the morphological changes in muscle tissue of four children with infantile Pompe's disease who received recombinant human alpha-glucosidase from rabbit milk for 72 weeks. The patients were 2.5-8 months of age at entry. Prior to treatment, all patients showed lysosomal glycogen storage in skeletal and smooth muscle cells, vascular endothelium, Schwann cells, and perineurium. The first response to treatment was noticed in vascular endothelium and in peripheral nerves after 12 weeks of treatment at an enzyme dose of 15-20 mg/kg. Increasing the dose to 40 mg/kg led, after 72 weeks of treatment, to a reduction of glycogen storage and substantial improvement of muscle architecture in the least affected patient. Not all patients responded equally well, possibly due to differences in degree of glycogen storage and concomitant muscle pathology at the start of treatment. We conclude that intravenous administration of recombinant human alpha-glucosidase from rabbit milk can improve muscle morphology in classic infantile Pompe's disease when treatment is started before irreversible damage has occurred.

  15. p.[G576S; E689K]: pathogenic combination or polymorphism in Pompe disease?

    Science.gov (United States)

    Kroos, Marian A; Mullaart, Reinier A; Van Vliet, Laura; Pomponio, Robert J; Amartino, Hernan; Kolodny, Edwin H; Pastores, Gregory M; Wevers, Ron A; Van der Ploeg, Ans T; Halley, Dicky J J; Reuser, Arnold J J

    2008-08-01

    We discuss four cases of acid alpha-glucosidase deficiency (EC, 3.2.1.3/20) without evident symptoms of Pompe disease (OMIM No 232300) in individuals of Asian descent. In three cases, the deficiency was associated with homozygosity for the sequence variant c.[1726G>A; 2065G>A] in the acid alpha-glucosidase gene (GAA) translating into p.[G576S; E689K]. One of these cases was a patient with profound muscular atrophy, another had cardio-myopathy and the third had no symptoms. The fourth case, the mother of a child with Pompe disease, was compound heterozygote for the GAA sequence variants c.[1726G>A; 2065G>A]/c.2338G>A (p.W746X) and had no symptoms either. Further investigations revealed that c.[1726A; 2065A] is a common GAA allele in the Japanese and Chinese populations. Our limited study predicts that approximately 4% of individuals in these populations are homozygote c.[1726A; 2065A]. The height of this figure in contrast to the rarity of Pompe disease in Asian populations and the clinical history of the cases described in this paper virtually exclude that homozygosity for c.[1726A; 2065A] causes Pompe disease. As c.[1726A; 2065A] homozygotes have been observed with similarly low acid alpha-glucosidase activity as some patients with Pompe disease, we caution they may present as false positives in newborn screening programs especially in Asian populations.

  16. The natural course of non-classic Pompe's disease; a review of 225 published cases.

    Science.gov (United States)

    Winkel, Léon P F; Hagemans, Marloes L C; van Doorn, Pieter A; Loonen, M Christa B; Hop, Wim J C; Reuser, Arnold J J; van der Ploeg, Ans T

    2005-08-01

    Pompe's disease is a neuromuscular disorder caused by deficiency of lysosomal acid alpha-glucosidase. Recombinant human alpha- glucosidase is under evaluation as therapeutic drug. In light of this development we studied the natural course of cases not fitting the definition of classic infantile Pompe's disease. Our review of 109 reports including 225 cases shows a continuous spectrum of phenotypes. The onset of symptoms ranged from 0 to 71 years. Based on the available literature, no criteria to delineate clinical sub-types could be established.A common denominator of these cases is that first symptoms were related to or caused by muscle weakness. In general, patients with a later onset of symptoms seemed to have a better prognosis. Respiratory failure was the most frequent cause of death. CK, LDH, ASAT, ALAT and muscle glycogen levels were frequently but not always elevated. In most cases a muscle biopsy revealed lysosomal pathology, but normal muscle morphology does not exclude Pompe's disease. In 10% of the cases in which the enzyme assay on leukocytes was used, a normal alpha-glucosidase activity was reported. Data on skeletal muscle strength and function, pulmonary function, disability, handicap and quality of life were insufficiently reported in the literature. Studies of non-classic Pompe's disease should focus on these aspects, before enzyme replacement therapy becomes generally available.

  17. N-alkylated nitrogen-in-the-ring sugars: conformational basis of inhibition of glycosidases and HIV-1 replication.

    Science.gov (United States)

    Asano, N; Kizu, H; Oseki, K; Tomioka, E; Matsui, K; Okamoto, M; Baba, M

    1995-06-23

    The conformations of nitrogen-in-the-ring sugars and their N-alkyl derivatives were studied from 1H NMR analyses, mainly using 3J(H,H) coupling constants and quantitative NOE experiments. No significant difference was seen in the ring conformation of 1-deoxynojirimycin (1), N-methyl-1-deoxynojirimycin (2), and N-butyl-1-deoxynojirimycin (3). However, it was shown that the C6 OH group in 1 is predominantly equatorial to the piperidine ring, while that in 2 or 3 is predominantly axial, and its N-alkyl group is oriented equatorially. In the furanose analogues 1,4-dideoxy-1,4-imino-D-arabinitol (4) and its N-methyl (5) and N-butyl (6) derivatives, the five-membered ring conformation differed significantly by the presence or absence of the N-substituted group and the length of the N-alkyl chain. Compound 3 reduced its inhibitory effect on almost all glycosidases, resulting in an extremely specific inhibitor for processing alpha-glucosidase I since N-alkylation of 1 is known to enhance both the potency and specificity of this enzyme in vitro and in vivo. This preferred (C6 OH axial) conformation in 2 and 3 appears to be responsible for their strong alpha-glucosidase I activity. Compound 4 is a good inhibitor of intestinal alpha-glucohydrolases, alpha-glucosidase II, and Golgi alpha-mannosidases I and II, but its N-alkyl derivatives 5 and 6 markedly decreased inhibitory potential for all enzymes tested. In the case of 2,5-dideoxy-2,5-imino-D-mannitol (DMDP, 7), which is a potent beta-galactosidase inhibitor, its N-methyl (8) and N-butyl (9) derivatives completely lost potency toward beta-galactosidase as well. N-Alkylation of compounds 4 and 7, known well as potent yeast alpha-glucosidase inhibitors, resulted in a serious loss of inhibitory activity toward yeast alpha-glucohydrolases. Activity of these nine analogues against HIV-1 replication was determined, based on the inhibition of virus-induced cytopathogenicity in MT-4 and MOLT-4 cells. Compounds 2 and 3, which are

  18. In vitro inhibition activity of polyphenol-rich extracts from Syzygium aromaticum(L.)Merr.& Perry(Clove)buds against carbohydrate hydrolyzing enzymes linked to type 2 diabetes and Fe2+-induced lipid peroxidation in rat pancreas

    Institute of Scientific and Technical Information of China (English)

    Stephen; Adeniyi; Adefegha; Ganiyu; Oboh

    2012-01-01

    Objective:To investigate and compare the inhibitor)’properties)of free and bound phenolic extracts of clove bud against carbohydrate hydrolyzing enzymes(alpha-amylase&alphaglucosidase)and Fe2+-induced lipid peroxidation in rat pancreas in vitro.Methods:The free phenolics were extracted with 80%.(v/v)acetone,while bound phenolics were extracted from the alkaline and acid hydrolyzed residue with ethyl acetate.Then,the interaction of the extracts with alpha-amylase and alpha-glucosidase was subsequently assessed.Thereafter,the total phenolic contents and antioxidant activities of the extracts were determined.Results:The result revealed that both extracts inhibited alpha-amylase and alpha-glucosidase in a dose-dependent manner.However,the alpha-glucosidase inhibitory activity of the extracts were significantly(P<0.05)higher than their alpha-amylase inhibitory activity.The free phenolics(31.67 mg/g)and flavonoid(17.28 mg/g)contents were significantly(P<0.05)higher than bound phenolic(23.52 mg/g)and flavonoid(13.70 mg/g)contents.Both extracts also exhibited high antioxidant activities as typified by their high reducing power,LI diphenyl-2-picrylhydrazyl(DPPH)and 2,2-azinobis-3-ethylbenzo-thiazoline-6-sulfonate(ABTS)radical scavenging abilities,as well as inhibition of Fe2+-induced lipid peroxidation in rat pancreas in vitro.Conclusions:This study provides a biochemical rationale by which clove elicits therapeutic effect on type 2 diabetes.

  19. Alfa-glucosidase-inhibiting activity of some Mexican plants used in the treatment of type 2 diabetes.

    Science.gov (United States)

    Andrade-Cetto, Adolfo; Becerra-Jiménez, Jaime; Cárdenas-Vázquez, René

    2008-02-28

    Type 2 diabetes is an endocrine disease, which accounts for 9% of deaths worldwide. The aim of oral therapy is to reach normoglycemia to prevent later complications. Among glucose-lowering medications, alpha-glucosidase inhibitors delay the absorption of ingested carbohydrates, reducing the postprandial glucose and insulin peaks. In the present study, we tested the butanolic extracts of four Mexican plants with respect to their alpha-glucosidase inhibition activity, without excluding other possible mechanisms of action. The plants Cecropia obtusifolia Bertol., Equisetum myriochaetum Schlecht & Cham, Acosmium panamense (Benth.) Yacolev and Malmea depressa (Baill) R.E. Fries are used in traditional medicine to treat type 2 diabetes. In previous studies, we have demonstrated these plants' hypoglycemic activity and determined the phytochemical composition of their extracts. Our results in n-STZ diabetic rats loaded with maltose showed that Malmea and Acosmium extracts decreased plasma glucose significantly from 30 min on resembling the effect of acarbose. Cecropia extract produced the highest reduction of plasma glucose, and at 90 min, the glucose level was lower than the fasting level, which suggests another mechanism of action. Equisetum did not exert any effect. In vitro assays of alpha-glucosidase activity showed an IC(50) of 14 microg/ml for Cecropia, 21 microg/ml for Malmea, and 109 microg/ml for Acosmium, which were lower than that of acarbose (128 microg/ml). Equisetum did not show any significant effect on this assay, either. These results contribute to understand the mechanism of action of these plants on glucose metabolism.

  20. A cross-sectional single-centre study on the spectrum of Pompe disease, German patients: molecular analysis of the GAA gene, manifestation and genotype-phenotype correlations

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    Herzog Andreas

    2012-06-01

    Full Text Available Abstract Background Pompe disease (Glycogen storage disease type II, GSD II, acid alpha-glucosidase deficiency, acid maltase deficiency, OMIM # 232300 is an autosomal-recessive lysosomal storage disorder due to a deficiency of acid alpha-glucosidase (GAA, acid maltase, EC 3.2.1.20, Swiss-Prot P10253. Clinical manifestations are dominated by progressive weakness of skeletal muscle throughout the clinical spectrum. In addition, the classic infantile form is characterised by hypertrophic cardiomyopathy. Methods In a cross-sectional single-centre study we clinically assessed 3 patients with classic infantile Pompe disease and 39 patients with non-classic presentations, measured their acid alpha-glucosidase activities and analysed their GAA genes. Results Classic infantile patients had nearly absent residual enzyme activities and a typical clinical course with hypertrophic cardiomyopathy until the beginning of therapy. The disease manifestations in non-classic patients were heterogeneous. There was a broad variability in the decline of locomotive and respiratory function. The age of onset ranged from birth to late adulthood and correlated with enzyme activities. Molecular analysis revealed as many as 33 different mutations, 14 of which are novel. All classic infantile patients had two severe mutations. The most common mutation in the non-classic group was c.-32-13 T > G. It was associated with a milder course in this subgroup. Conclusions Disease manifestation strongly correlates with the nature of the GAA mutations, while the variable progression in non-classic Pompe disease is likely to be explained by yet unknown modifying factors. This study provides the first comprehensive dataset on the clinical course and the mutational spectrum of Pompe disease in Germany.

  1. Nitrogen-in-the-ring pyranoses and furanoses: structural basis of inhibition of mammalian glycosidases.

    Science.gov (United States)

    Asano, N; Oseki, K; Kizu, H; Matsui, K

    1994-10-28

    Seven pyranoses and three furanoses with a nitrogen in the ring were prepared by chemical synthesis, microbial conversion, and isolation from plants to investigate the contribution of epimerization, deoxygenation, and conformation to the potency of inhibition and specificity of mammalian glycosidases. The seven pyranoses are 1-deoxynojirimycin (1), the D-manno (2), D-allo (3), and D-galacto (4) isomers of 1, fagomine (1,2-dideoxynojirimycin, 5), and the D-allo (6) and D-galacto (7) isomers of 5, while the three furanoses are 2,5-dideoxy-2,5-imino-D-mannitol (8), 1,4-dideoxy-1,4-imino-D-arabinitol (9), and 1,4-dideoxy-1,4-imino-D-ribitol (10). The 2-deoxygenation and/or 3-epimerization of 1 enhanced the potency for rat intestinal lactase and bovine liver cytosolic beta-galactosidase. Especially compound 6 showed a potent inhibitory activity against both enzymes, and compound 8, a mimic of beta-D-fructofuranose, was a potent inhibitor of both beta-galactosidases as well. Compound 4, which has been known as a powerful alpha-galactosidase inhibitor, exhibited no significant inhibitory activity for most of mammalian beta-galactosidases. In addition, compound 6 fairly retained a potency of 1 toward rat intestinal isomaltase. In this study, compound 8, known as a processing alpha-glucosidase I inhibitor in cell culture, has been found to have no effect on processing alpha-glucosidase II, whereas 9 has been shown to be a good nonspecific inhibitor of intestinal isomaltase, processing alpha-glucosidase II, Golgi alpha-mannosidases I and II, and porcine kidney trehalase. It has been speculated that glycosidase inhibitors have structures which resemble those of the respective glycosyl cations. This Broad inhibitory activity of 9 toward various glycosidases suggest that it superimposes well on the various glycosyl cations.

  2. Adult onset glycogen storage disease type II (adult onset Pompe disease): report and magnetic resonance images of two cases

    Energy Technology Data Exchange (ETDEWEB)

    Del Gaizo, Andrew [Emory University School of Medicine, Radiology Resident, Atlanta, GA (United States); Banerjee, Sima [Emory University School of Medicine, Musculoskeletal Radiology Department, Atlanta, GA (United States); Terk, Michael [Emory University School of Medicine, Radiology, Division of Musculoskeletal Imaging, Atlanta, GA (United States)

    2009-12-15

    Glycogen storage disease type II (GSDII), also referred to as Pompe disease or acid maltase deficiency, is a rare inherited condition caused by a deficiency in acid alpha-glucosidase (GAA) enzyme activity. The condition is often classified by age of presentation, with infantile and late onset variants (Laforet et al. J Neurology 55:1122-8, 2000). Late onset tends to present with progressive proximal muscle weakness and respiratory insufficiency (Winkel et al. J Neurology 252:875-84, 2005). We report two cases of biopsy confirmed adult onset GSDII, along with key Magnetic Resonance (MR) images. (orig.)

  3. Infantile Onset Glycogen Storage Disease Type 2: Case Report

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    Serkan Bilge Koca

    2014-08-01

    Full Text Available Glycogen storage disease type 2 (Pompe’s disease is an autosomal recessive, fatal glycogen storage disease presenting with hypotonia and muscle weakness. It is known that deficiency of lysosomal acid alpha-glucosidase (acid maltase leads to progressive generalised myopathy, cardiomyopathy and death in early infancy because of respiratory muscle weakness. Excessive undegradable intracellular glycogen deposition plays a role in the pathogenesis of the disease. Here we report a 3.5 month-old girl presenting with respiratory failure due to pneumonia and hypotonia, who was later diagnosed as Pompe disease.

  4. Concurrent Therapy with a Low-carbohydrate Diet and Miglitol Remarkably Improved the Postprandial Blood Glucose and Insulin Levels in a Patient with Reactive Hypoglycemia due to Late Dumping Syndrome.

    Science.gov (United States)

    Hirose, Sachie; Iwahashi, Yasuyuki; Seo, Akane; Sumiyoshi, Michitaka; Takahashi, Tetsuya; Tamori, Yoshikazu

    2016-01-01

    Reactive hypoglycemia induced by late dumping syndrome is often observed after gastrectomy. However, no effective therapy has yet been fully established. We herein describe a case in which concurrent therapy with a low-carbohydrate diet using low-glycemic-index food and an alpha-glucosidase inhibitor, miglitol, very effectively ameliorated the postprandial fluctuations in the blood glucose and plasma insulin levels in a patient with reactive hypoglycemia due to late dumping syndrome following total gastrectomy. The administration of miglitol under a low-carbohydrate diet using low-glycemic-index food may therefore be an ideal treatment for reactive hypoglycemia due to late dumping syndrome.

  5. Testudinibacter aquarius gen. nov., sp nov., a member of the family Pasteurellaceae isolated from the oral cavity of freshwater turtles

    DEFF Research Database (Denmark)

    Hansen, Mie Johanne; Pennanen, Elin Anna Erica; Bojesen, Anders Miki

    2016-01-01

    Pasteurellaceae. However, they could be separated from existing genera of the Pasteurellaceae by the following test results: indole, ornithine decarboxylase and Voges-Proskauer positive; and methyl red, urease and PNPG (alpha-glucosidase) negative. No X- or V-factor requirement was observed. A zone of beta....... nov., sp. nov. The type strain of Testudinibacter aquarius is ELNT2x(T) (=CCUG 65146(T)=DSM 28140(T)), which was isolated from the oral cavity of a captive eastern long-necked turtle (Chelodina longicollis) in Denmark in 2012....

  6. REDUCTION OF CARDIOVASCULAR COMPLICATIONS OF MODERN HYPOGLYCEMIC THERAPY OF DIABETES MELLITUS TYPE 2: "FLORENTINE HERESY"

    Directory of Open Access Journals (Sweden)

    A. A. Aleksandrov

    2010-01-01

    Full Text Available The classic hypoglycemic agents include biguanides, sulfonylurea drugs, meglitinides, glitazones and alpha-glucosidase inhibitors. Modern algorithm of hypoglycemic therapy in the first step considers lifestyle modification and metformin monotherapy, the second step — the combined therapy. However, the effect of combined hypoglycemic therapy on long-term cardiovascular prognosis in patients with type 2 diabetes mellitus is studied insufficiently. Combined therapy with glibenclamide and metformin can result in adverse cardiovascular effects, so that long term therapy should be avoided in patients with coronary heart disease. Adequate pharmacological approaches to hyperglycemia correction should be elaborated.

  7. Evaluation of antihyperglycemia and antihypertension potential of native Peruvian fruits using in vitro models.

    Science.gov (United States)

    Pinto, Marcia Da Silva; Ranilla, Lena Galvez; Apostolidis, Emmanouil; Lajolo, Franco Maria; Genovese, Maria Inés; Shetty, Kalidas

    2009-04-01

    Local food diversity and traditional crops are essential for cost-effective management of the global epidemic of type 2 diabetes and associated complications of hypertension. Water and 12% ethanol extracts of native Peruvian fruits such as Lucuma (Pouteria lucuma), Pacae (Inga feuille), Papayita arequipeña (Carica pubescens), Capuli (Prunus capuli), Aguaymanto (Physalis peruviana), and Algarrobo (Prosopis pallida) were evaluated for total phenolics, antioxidant activity based on 2, 2-diphenyl-1-picrylhydrazyl radical scavenging assay, and functionality such as in vitro inhibition of alpha-amylase, alpha-glucosidase, and angiotensin I-converting enzyme (ACE) relevant for potential management of hyperglycemia and hypertension linked to type 2 diabetes. The total phenolic content ranged from 3.2 (Aguaymanto) to 11.4 (Lucuma fruit) mg/g of sample dry weight. A significant positive correlation was found between total phenolic content and antioxidant activity for the ethanolic extracts. No phenolic compound was detected in Lucuma (fruit and powder) and Pacae. Aqueous extracts from Lucuma and Algarrobo had the highest alpha-glucosidase inhibitory activities. Papayita arequipeña and Algarrobo had significant ACE inhibitory activities reflecting antihypertensive potential. These in vitro results point to the excellent potential of Peruvian fruits for food-based strategies for complementing effective antidiabetes and antihypertension solutions based on further animal and clinical studies.

  8. Physicochemical and biofunctional properties of crab chitosan nanoparticles.

    Science.gov (United States)

    Nguyen, The Han; Kwak, Hae Soo; Kim, Sang Moo

    2013-08-01

    The physicochemical and biofunctional properties of crab chitosan nanoparticles of two different sizes (Nano A and B) manufactured by dry milling method were evaluated for commercialization. The deacetylation degrees (DD) of Nano A, B and the control chitosan were 90.9, 93.0, and 92.7% respectively whereas their molecular weights (M(w)) were 43.9, 44.7 and 208.8 kDa. The average sizes of the dispersed Nano A, B and the control chitosan in cetyltrimethylammonium chloride were 735.9, 849.4 and 2,382.4 nm, respectively, which were lower than 1441.7, 2935.6 and 6832.9 nm of the intact chitosans. Chitosan nanoparticles had mild tyrosinase, antioxidant and angiotensin I converting enzyme (ACE), but weak collagenase, elastase and beta-glucuronidase inhibitory activity. However, Nano A had strong alpha-glucosidase inhibitory activity, which was comparable to that of acarbose, a commercial alpha-glucosidase inhibitor. In addition, the minimum inhibitory concentrations (MICs) of chitosan and its nanoparticles ranged from 30 to > 200 microg/mL against each four gram-positive and gram-negative bacteria. Therefore, crab chitosan nanoparticles could be used as a nutraceutical, cosmeceutical or pharmaceutical product.

  9. Pompe disease: clinical perspectives

    Directory of Open Access Journals (Sweden)

    Cabello JF

    2016-12-01

    Full Text Available Juan Francisco Cabello,1 Deborah Marsden21Genetics and Metabolic Disease Laboratory, Nutrition and Food Technology Institute (INTA, University of Chile, Santiago, Chile; 2Division of Genetics and Genomics, Boston Children's Hospital, Boston, MA, USA Abstract: Pompe disease (acid alpha-glucosidase deficiency, OMIM 232300 is a rare lysosomal storage disorder due to autosomal recessive mutations in the GAA gene. It has also been called acid maltase deficiency and glycogen storage disease type II. There is a broad clinical presentation: the most severe form that presents in the first few months of life with cardiomyopathy and generalized muscle weakness that rapidly progresses to death from cardio-respiratory failure in the first year of life (infant-onset Pompe disease. A more slowly progressive disease, with little or no cardiac involvement, presents with proximal myopathy and/or pulmonary insufficiency, from the second year of life to late adulthood (late-onset Pompe disease. The recent development and introduction of enzyme replacement therapy with intravenous infusion of recombinant human acid alpha-glucosidase have made a major improvement in the morbidity and mortality of this disease. New therapies are also in development. With the availability of treatment, diagnostic methods have also improved, allowing for earlier recognition and potential early therapeutic intervention. The advent of newborn screening for Pompe disease may identify patients who can be treated before significant irreversible disease has occurred. Keywords: Pompe disease, glycogen storage disease, lysosomal storage disease, enzyme replacement therapy, gene therapy, chaperone therapy, genotype/phenotype, newborn screening

  10. In-vitro α amylase and glycosidase inhibitory effect of ethanolic extract of antiasthmatic drug - Shirishadi

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    Divya Kajaria

    2013-01-01

    Full Text Available Asthma and diabetes have strong relationship; both are cause and effect of each other. Oxidative stress due to bronchial asthma may cause insulin resistance whereas lack of proper insulin can cause defective smooth muscle relaxant. There is no single medicine available that can manage both diseases, rather the mainstay treatment of bronchial asthma causes hyperglycemia. Keeping this problem in focus, in this study the hypoglycemic effect of an indigenous antiasthmatic Ayurvedic drug Shirishadi was evaluated. Pancreatic alpha amylase and glucosidase inhibitors offer an effective strategy to lower the level of post prandial hyperglycemia via control of starch breakdown. For evaluation of hypoglycemic activity of drug, in-vitro alpha amylase and alpha glucosidase enzyme inhibition was calculated. Ethanolic extract of compound showed 76.40% + 0.88% reduction in alpha amylase activity and 63.85% + 0.36% in alpha glucosidase activity with IC 50 0.68 mg/ml and 2.89 mg/ml, respectively. This study suggests that the ethanolic extract of Shirishadi polyherbal compound effectively acts as alpha amylase and glucosidase inhibitor leading to a reduction in starch hydrolysis and hence acts as antiasthmatic as well as hypoglycemic drug.

  11. Pompe's disease.

    Science.gov (United States)

    van der Ploeg, Ans T; Reuser, Arnold J J

    2008-10-11

    Pompe's disease, glycogen-storage disease type II, and acid maltase deficiency are alternative names for the same metabolic disorder. It is a pan-ethnic autosomal recessive trait characterised by acid alpha-glucosidase deficiency leading to lysosomal glycogen storage. Pompe's disease is also regarded as a muscular disorder, but the generalised storage of glycogen causes more than mobility and respiratory problems. The clinical spectrum is continuous and broad. First symptoms can present in infants, children, and adults. Cardiac hypertrophy is a key feature of classic infantile Pompe's disease. For a long time, there was no means to stop disease progression, but the approval of enzyme replacement therapy has substantially changed the prospects for patients. With this new development, the disease is now among the small but increasing number of lysosomal storage disorders, for which treatment has become a reality. This review is meant to raise general awareness, to present and discuss the latest insights in disease pathophysiology, and to draw attention to new developments about diagnosis and care. We also discuss the developments that led to the approval of enzyme replacement therapy with recombinant human alpha-glucosidase from Chinese hamster ovary cells (alglucosidase alfa) by the US Food and Drug Administration and European Medicines Agency in 2006, and review clinical practice.

  12. Update of the Pompe disease mutation database with 107 sequence variants and a format for severity rating.

    Science.gov (United States)

    Kroos, Marian; Pomponio, Robert J; van Vliet, Laura; Palmer, Rachel E; Phipps, Michael; Van der Helm, Robert; Halley, Dicky; Reuser, Arnold

    2008-06-01

    Pompe disease was named after the Dutch pathologist Dr JC Pompe who reported about a deceased infant with idiopathic hypertrophy of the heart. The clinical findings were failure to thrive, generalized muscle weakness and cardio-respiratory failure. The key pathologic finding was massive storage of glycogen in heart, skeletal muscle and many other tissues. The disease was classified as glycogen storage disease type II and decades later shown to be a lysosomal disorder caused by acid alpha-glucosidase deficiency. The clinical spectrum of Pompe disease appeared much broader than originally recognized. Adults with the same enzyme deficiency, alternatively named acid maltase deficiency, were reported to have slowly progressive skeletal muscle weakness and respiratory problems, but no cardiac involvement. The clinical heterogeneity is largely explained by the kind and severity of mutations in the acid alpha-glucosidase gene (GAA), but secondary factors, as yet unknown, have a substantial impact. The Pompe disease mutation database aims to list all GAA sequence variations and describe their effect. This update with 107 sequence variations (95 being novel) brings the number of published variations to 289, the number of non-pathogenic mutations to 67 and the number of proven pathogenic mutations to 197. Further, this article introduces a tool to rate the various mutations by severity, which will improve understanding of the genotype-phenotype correlation and facilitate the diagnosis and prognosis in Pompe disease.

  13. Enzyme analysis for Pompe disease in leukocytes; superior results with natural substrate compared with artificial substrates.

    Science.gov (United States)

    van Diggelen, O P; Oemardien, L F; van der Beek, N A M E; Kroos, M A; Wind, H K; Voznyi, Y V; Burke, D; Jackson, M; Winchester, B G; Reuser, A J J

    2009-06-01

    Enzyme analysis for Pompe disease in leukocytes has been greatly improved by the introduction of acarbose, a powerful inhibitor of interfering alpha-glucosidases, which are present in granulocytes but not in lymphocytes. Here we show that the application of acarbose in the enzymatic assay employing the artificial substrate 4-methylumbelliferyl-alpha-D: -glucoside (MU-alphaGlc) is insufficient to clearly distinguish patients from healthy individuals in all cases. Also, the ratios of the activities without/with acarbose only marginally discriminated Pompe patients and healthy individuals. By contrast, when the natural substrate glycogen is used, the activity in leukocytes from patients (n = 82) with Pompe disease is at most 17% of the lowest control value. The use of artificial substrate in an assay with isolated lymphocytes instead of total leukocytes is a poor alternative as blood samples older than one day invariably yield lymphocyte preparations that are contaminated with granulocytes. To diagnose Pompe disease in leukocytes we recommend the use of glycogen as substrate in the presence of acarbose. This assay unequivocally excludes Pompe disease. To also exclude pseudo-deficiency of acid alpha-glucosidase caused by the sequence change c.271G>A (p.D91N or GAA2; homozygosity in approximately 1:1000 caucasians), a second assay employing MU-alphaGlc substrate plus acarbose or DNA analysis is required.

  14. 6,7-diepicastanospermine, a tetrahydroxyindolizidine alkaloid inhibitor of amyloglucosidase

    Energy Technology Data Exchange (ETDEWEB)

    Molyneux, R.J.; Benson, M. (Dept. of Agriculture, Albany, CA (United States)); Pan, Y.T.; Tropea, J.E.; Kaushal, G.P.; Elbein, A.D. (Univ. of Texas, San Antonio (United States))

    1991-10-15

    A tetrahydroxyindolizidine alkaloid, 6,7-diepicastanospermine, was isolated from the seeds of Castanospermum australe by extraction with methanol and purified to homogeneity using ion-exchange, preparative thin-layer, and radial chromatography. A very low yield of a pyrrolidine alkaloid, N-(hydroxyethyl)-2-(hydroxymethyl)-3-hydroxypyrrolidine, was also obtained by analogous methods. The purity of both alkaloids was established by gas chromatography of their trimethylsilyl (TMS) derivatives as better than 99%. The molecular weight of each alkaloid was established as 189 and 161, respectively, by mass spectrometry, and the structure of each was deduced from their {sup 1}H and {sup 13}C NMR spectra. The structure of the pyrrolidine alkaloids which co-occur in C. australe. 6,7-Diepicastanospermine was found to be a moderately good inhibitor of the fungal {alpha}-glucosidase, amyloglucosidase and a relatively weak inhibitor of {beta}-glucosidase. It failed to inhibit {alpha}-glucosidase. It failed to inhibit {alpha}- or {beta}-galactosidase, {alpha}- or {beta}-mannosidase, or {alpha}-L-fucosidase. Comparison of its inhibitory activity toward amyloglucosidase with those of its isomers, castanospermine and 6-epicastanospermine, demonstrated that epimerization of a single hydroxyl group can produce significant alteration of such inhibitory properties.

  15. A complex craniovertebral junction malformation in a patient with late onset glycogenosis 2

    Directory of Open Access Journals (Sweden)

    Mariasofia Cotelli

    2014-01-01

    Full Text Available Glycogenosis II (GSDII is an autosomal recessive lysosomal storage disorder resulting from deficiency of acid alpha-glucosidase and subsequent lysosomal accumulation of glycogen in skeletal, cardiac and smooth muscles. The late-onset form is characterized by wide variability of the phenotypical spectrum. Clinical findings may include muscle weakness, respiratory insufficiency, vascular abnormalities, low bone mineral density and higher risk of developing osteoporosis. Craniovertebral junction (CVJ malformations have never been described so far. We here report on a GSDII 43-year-old woman who harbored the mutations IVS1-13T>G and c.2237G>A in the acid alpha-glucosidase gene. She recurrently suffered from headache, neck pain and dizziness. Brain MRI and CT scan showed the presence of a very rare complex CVJ malformation composed of basilar invagination, basiocciput hypoplasia, partial C1 assimilation, C1 posterior arch aplasia and C1 lateral mass hypoplasia and offset. Although we cannot rule out their coincidental occurrence, the rarity of multiple CVJ malformations in the general population as well as the well-known GSDII multisystem involvement should suggest to study the CVJ in the diagnostic process of GSDII patients in order to assess the CVJ malformation frequency in GSDII population and verify a possible relationship between these two conditions.

  16. The sim operon facilitates the transport and metabolism of sucrose isomers in Lactobacillus casei ATCC 334.

    Science.gov (United States)

    Thompson, John; Jakubovics, Nicholas; Abraham, Bindu; Hess, Sonja; Pikis, Andreas

    2008-05-01

    Inspection of the genome sequence of Lactobacillus casei ATCC 334 revealed two operons that might dissimilate the five isomers of sucrose. To test this hypothesis, cells of L. casei ATCC 334 were grown in a defined medium supplemented with various sugars, including each of the five isomeric disaccharides. Extracts prepared from cells grown on the sucrose isomers contained high levels of two polypeptides with M(r)s of approximately 50,000 and approximately 17,500. Neither protein was present in cells grown on glucose, maltose or sucrose. Proteomic, enzymatic, and Western blot analyses identified the approximately 50-kDa protein as an NAD(+)- and metal ion-dependent phospho-alpha-glucosidase. The oligomeric enzyme was purified, and a catalytic mechanism is proposed. The smaller polypeptide represented an EIIA component of the phosphoenolpyruvate-dependent sugar phosphotransferase system. Phospho-alpha-glucosidase and EIIA are encoded by genes at the LSEI_0369 (simA) and LSEI_0374 (simF) loci, respectively, in a block of seven genes comprising the sucrose isomer metabolism (sim) operon. Northern blot analyses provided evidence that three mRNA transcripts were up-regulated during logarithmic growth of L. casei ATCC 334 on sucrose isomers. Internal simA and simF gene probes hybridized to approximately 1.5- and approximately 1.3-kb transcripts, respectively. A 6.8-kb mRNA transcript was detected by both probes, which was indicative of cotranscription of the entire sim operon.

  17. An enzymatic method for the determination of fructans in foods and food products - Comparison of the results by high performance anion exchange chromatography with pulsed amperemetric detection

    DEFF Research Database (Denmark)

    Andersen, Rikke; Sørensen, A.

    1999-01-01

    effects than the fructanase reported as normally used. The method is tested on ten standard substances and five fructan products, and nine foods and food products are also analysed. The enzymatic measurement of the released sugars is confirmed by measurements done by high performance anion exchange......We report a new and non-equipment demanding method of measuring the content of fructans as well as the contents of free glucose, free fructose and sucrose in foods and food products enzymatically. This method comprises hydrolysis of fructans into D-glucose and D-fructose enzymatically...... and measurement of the released sugars enzymatically. Sucrose is hydrolysed by alpha-glucosidase instead of beta-fructosidase, which is normally used. In addition, sucrose is measured in the form of D-fructose instead of the typical D-glucose form, and the fructanase used to hydrolyse the fructans has fewer side...

  18. New Enzyme-Inhibitory Triterpenoid from Marine Macro Brown Alga Padina boergesenii Allender & Kraft

    Science.gov (United States)

    Ali, Liaqat; Khan, Abdul Latif; Al-Broumi, Muhammad; Al-Harrasi, Rashid; Al-Kharusi, Lubna; Hussain, Javid; Al-Harrasi, Ahmed

    2017-01-01

    In continuation to our study of the chemical and biological potential of the secondary metabolites isolated from Omani seaweeds, we investigated a marine brown alga, Padina boergesenii. The phytochemical investigation resulted in the isolation of a new secondary metabolite, padinolic acid (1), along with some other semi-pure fractions and sub-fractions. The planar structure was confirmed through MS and NMR (1D and 2D) spectral data. The NOESY experiments coupled with the biogenetic consideration were helpful in assigning the stereochemistry in the molecule. Compound 1 was subjected to enzyme inhibition studies using urease, lipid peroxidase, and alpha-glucosidase enzymes. Compound 1 showed low to moderate α-glucosidase and urease enzyme inhibition, respectively, and moderate anti-lipid peroxidation activities. The current study indicates the potential of this seaweed and provides the basis for further investigation. PMID:28106757

  19. New Enzyme-Inhibitory Triterpenoid from Marine Macro Brown Alga Padina boergesenii Allender & Kraft

    Directory of Open Access Journals (Sweden)

    Liaqat Ali

    2017-01-01

    Full Text Available In continuation to our study of the chemical and biological potential of the secondary metabolites isolated from Omani seaweeds, we investigated a marine brown alga, Padina boergesenii. The phytochemical investigation resulted in the isolation of a new secondary metabolite, padinolic acid (1, along with some other semi-pure fractions and sub-fractions. The planar structure was confirmed through MS and NMR (1D and 2D spectral data. The NOESY experiments coupled with the biogenetic consideration were helpful in assigning the stereochemistry in the molecule. Compound 1 was subjected to enzyme inhibition studies using urease, lipid peroxidase, and alpha-glucosidase enzymes. Compound 1 showed low to moderate α-glucosidase and urease enzyme inhibition, respectively, and moderate anti-lipid peroxidation activities. The current study indicates the potential of this seaweed and provides the basis for further investigation.

  20. Oleanolic acid and related derivatives as medicinally important compounds.

    Science.gov (United States)

    Sultana, Nighat; Ata, Athar

    2008-12-01

    Oleanolic acid has been isolated from chloroform extract of Olea ferruginea Royle after removal of organic bases and free acids. The literature survey revealed it to be biologically very important. In this review the biological significance of oleanolic acid and its derivatives has been discussed. The aim of this review is to update current knowledge on oleanolic acid and its natural and semisynthetic analogs, focussing on its cytotoxic, antitumer, antioxidant, anti-inflamatory, anti-HIV, acetyl cholinesterase, alpha-glucosidase, antimicrobial, hepatoprotective, anti-inflammatory, antipruritic, spasmolytic activity, anti-angiogenic, antiallergic, antiviral and immunomodulatory activities. We present in this review, for the first time, a compilation of the most relevant scientific papers and technical reports of the chemical, pre-clinical and clinical research on the properties of oleanolic acid and its derivatives.

  1. Bacterial and enzymatic bioassays for toxicity testing in the environment

    Energy Technology Data Exchange (ETDEWEB)

    Bitton, G.; Koopman, B. (Department of Environmental Engineering Sciences, University of Florida, Gainesville (United States))

    1992-01-01

    Microbioassays using bacteria or enzymes are increasingly applied to measure chemical toxicity in the environment. Attractive features of these assays may include low cost, rapid response to toxicants, high sample throughput, modest laboratory equipment and space requirements, low sample volume, portability, and reproducible responses. Enzymatic tests rely on measurement of either enzyme activity or enzyme biosynthesis. Dehydrogenases are the enzymes most used in toxicity testing. Assay of dehydrogenase activity is conveniently carried out using oxidoreduction dyes such as tetrazolium salts. Other enzyme activity tests utilize ATPases, esterases, phosphatases, urease, luciferase, beta-galactosidase, protease, amylase, or beta-glucosidase. Recently, the inhibition of enzyme (beta-galactosidase, tryptophanase, alpha-glucosidase) biosynthesis has been explored as a basis for toxicity testing. Enzyme biosynthesis was found to be generally more sensitive to organic chemicals than enzyme activity.107 references.

  2. Modulation of P2 receptors on pancreatic β-cells by agonists and antagonists: a molecular target for type 2 diabetes treatment.

    Science.gov (United States)

    Pacheco, Paulo Anastácio Furtado; Ferreira, Leonardo Gomes Braga; Alves, Luiz Anastacio; Faria, Robson Xavier

    2013-05-01

    Morbidity and mortality from diabetes mellitus (DM) are serious worldwide concerns. By the year 2030, the estimated number of diabetic patients will reach a staggering 439 million worldwide. Diabetes mellitus type 2 (DM2), which involves disturbances in both insulin secretion and resistance, is the most common form of diabetes and affects approximately 5 to 7% of the world's population. When a patient with DM2 cannot regulate his or her blood glucose levels through diet, weight loss, or exercise, oral medications, such as hypoglycemic agents (i.e., sulphonylureas, biguanides, alpha glucosidase inhibitors and thiazolidinediones), are crucial. Here, we discuss some physiological aspects of P2 receptors on pancreatic β-cells, which express a variety of P2 receptor isoforms. These receptors enhance glucose-dependent insulin release. In addition, we speculate on the potential of purinergic compounds as novel or additional treatments for Type 2 Diabetes mellitus.

  3. New Enzyme-Inhibitory Triterpenoid from Marine Macro Brown Alga Padina boergesenii Allender & Kraft.

    Science.gov (United States)

    Ali, Liaqat; Khan, Abdul Latif; Al-Broumi, Muhammad; Al-Harrasi, Rashid; Al-Kharusi, Lubna; Hussain, Javid; Al-Harrasi, Ahmed

    2017-01-18

    In continuation to our study of the chemical and biological potential of the secondary metabolites isolated from Omani seaweeds, we investigated a marine brown alga, Padina boergesenii. The phytochemical investigation resulted in the isolation of a new secondary metabolite, padinolic acid (1), along with some other semi-pure fractions and sub-fractions. The planar structure was confirmed through MS and NMR (1D and 2D) spectral data. The NOESY experiments coupled with the biogenetic consideration were helpful in assigning the stereochemistry in the molecule. Compound 1 was subjected to enzyme inhibition studies using urease, lipid peroxidase, and alpha-glucosidase enzymes. Compound 1 showed low to moderate α-glucosidase and urease enzyme inhibition, respectively, and moderate anti-lipid peroxidation activities. The current study indicates the potential of this seaweed and provides the basis for further investigation.

  4. Opposing effects of dietary protein and sugar regulate a transcriptional target of Drosophila insulin-like peptide signaling.

    Science.gov (United States)

    Buch, Susanne; Melcher, Christoph; Bauer, Matthias; Katzenberger, Joerg; Pankratz, Michael J

    2008-04-01

    Specific neurosecretory cells of the Drosophila brain express insulin-like peptides (dilps), which regulate growth, glucose homeostasis, and aging. Through microarray analysis of flies in which the insulin-producing cells (IPCs) were ablated, we identified a target gene, target of brain insulin (tobi), that encodes an evolutionarily conserved alpha-glucosidase. Flies with lowered tobi levels are viable, whereas tobi overexpression causes severe growth defects and a decrease in body glycogen. Interestingly, tobi expression is increased by dietary protein and decreased by dietary sugar. This pattern is reminiscent of mammalian glucagon secretion, which is increased by protein intake and decreased by sugar intake, suggesting that tobi is regulated by a glucagon analog. tobi expression is also eliminated upon ablation of neuroendocrine cells that produce adipokinetic hormone (AKH), an analog of glucagon. tobi is thus a target of the insulin- and glucagon-like signaling system that responds oppositely to dietary protein and sugar.

  5. Identification of aphid salivary proteins: a proteomic investigation of Myzus persicae.

    Science.gov (United States)

    Harmel, N; Létocart, E; Cherqui, A; Giordanengo, P; Mazzucchelli, G; Guillonneau, F; De Pauw, E; Haubruge, E; Francis, F

    2008-04-01

    The role of insect saliva in the first contact between an insect and a plant is crucial during feeding. Some elicitors, particularly in insect regurgitants, have been identified as inducing plant defence reactions. Here, we focused on the salivary proteome of the green peach aphid, Myzus persicae. Proteins were either directly in-solution digested or were separated by 2D SDS-PAGE before trypsin digestion. Resulting peptides were then identified by mass spectrometry coupled with database investigations. A homemade database was constituted of expressed sequence tags from the pea aphid Acyrtosiphon pisum and M. persicae. The databases were used to identify proteins related to M. persicae with a nonsequenced genome. This procedure enabled us to discover glucose oxidase, glucose dehydrogenase, NADH dehydrogenase, alpha-glucosidase and alpha-amylase in M. persicae saliva. The presence of these enzymes is discussed in terms of plant-aphid interactions.

  6. Expanded bed chromatography of proteins in small-diameter columns. II. Methods development and scale up.

    Science.gov (United States)

    Ghose, S; Chase, H

    2000-01-01

    The scaled down system developed in Part I of this series was further validated by using a 1-cm diameter column for method development studies for the separation of two model proteins, alcohol dehydrogenase and alpha-glucosidase, from unclarified yeast homogenate by hydrophobic interaction expanded bed chromatography based on the STREAMLINE matrix. The efficacy of solids removal and establishment of optimal binding and separation condition by stepwise elution were investigated. Equilibration of the EBA column and loading at high salt strengths affected the subsequent recovery of the two target proteins. Although good resolution between the target proteins could be achieved, peak tailing was found to be a consistent problem. The optimised separation protocol was scaled up 25-fold to a column diameter of 5.0 cm. The results were in good agreement with the run conducted in the 1-cm column, indicating the potential of using the small columns as an viable approach for method scouting and development studies.

  7. SGLT 2 Inhibitors: A New Therapeutic Target And Its Role In Current Clinical Practice

    Directory of Open Access Journals (Sweden)

    PV Shiji

    2015-10-01

    Full Text Available Diabetes, one of the major life style diseases, is associated with high morbidity and mortality owing to its microvascular and macrovascular complications. The chance of development of various complications can be effectively prevented by tight glycemic control. We have various groups of drugs like Biguanides, Sulfonyl ureas, Glitazones, Alpha-glucosidase inhibitors, Incretin based therapy, Insulin and Insulin analogues in the armamentarium to treat diabetes. But still, the number of patients attaining glycemic targets are relatively low and various adverse effect limit the use of some of these drugs, especially in special groups. Hence there is ongoing research to develop newer and newer drugs which provide sustained blood glucose reduction with minimal adverse effects. SGLT-2 inhibitors are a new group of drugs recently approved by FDA to treat Diabetes. In this review we discuss about mechanism of action, various adverse effects and the clinical role of various SGLT-2 Inhibitors.

  8. Newer Approaches In The Treatment of Diabetes Mellitus

    Directory of Open Access Journals (Sweden)

    Kamlesh Patel

    2013-01-01

    Full Text Available Diabetes Mellitus is a public health problem worldwide. The most effective anti-diabetic drugs currently available include insulin and newer insulin preparations, sulphonylureas, biguanides, meglitinides, thiazolidinediones, alpha- glucosidase inhibitors, incretins, guargum and glucomannan. However, the future therapies will need to focus on those patents who do not respond well to these treatments and who account for 50% of the health care costs of diabetes mellitus. Drug development for diabetes mellitus has been directed at improving currently available drugs and findings new compounds. In this review article, we will review the role of future new chemical entities able to target the metabolic disorder. Some of these new anti-diabetic treatment strategies may in the future not only control symptoms and modify the natural course of diabetes, but also potentially prevent or cure the disease.

  9. Diet and the Role of Altered Carbohydrate Absorption in the Treatment of Noninsulin-Dependent Diabetes Mellitus

    Directory of Open Access Journals (Sweden)

    Thomas MS Wolever

    1996-01-01

    Full Text Available The gastrointestinal tract has no clear role in the pathophysiology of noninsulin-dependent diabetes mellitus (NIDDM, but it may be an appropriate site for therapeutic intervention, specifically changes in diet, meal frequency and medications. Studies suggest that for patients with NIDDM, a calorie-restricted, high carbohydrate diet low in fat and rich in fibre may improve glycemic control, mitigate the risk of atherosclerosis and retard such diabetic complications as nephropathy and retinopathy. Increased meal frequency slows the rate of carbohydrate absorption, flattens blood insulin responses and reduces serum cholesterol. New therapeutic interventions, such as soluble fibre, low glycemic index foods or alpha glucosidase inhibitors, can further slow carbohydrate absorption and thus reduce secondary risks from hyperglycemia and hyperinsulinemia.

  10. Dicty_cDB: SHE417 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ne) Homo sapiens sucrase-isomaltase (a... 78 1e-13 BC116452_1( BC116452 |pid:none) Homo sapiens sucrase-isomalt...ase (a... 78 1e-13 ( P14410 ) RecName: Full=Sucrase-isomaltase, intestinal; Contains:... 78 1e-13 M22616_...1( M22616 |pid:none) Human sucrase-isomaltase mRNA, partial... 78 1e-13 A36690( A...36690 ) sucrose alpha-glucosidase (EC 3.2.1.48) - rat (fr... 75 6e-13 L25926_1( L25926 |pid:none) Rat sucrase-isomalt..... 72 7e-12 ( P23739 ) RecName: Full=Sucrase-isomaltase, intestinal; Contains:...

  11. Disruption of Brewers' yeast by hydrodynamic cavitation: Process variables and their influence on selective release.

    Science.gov (United States)

    Balasundaram, B; Harrison, S T L

    2006-06-01

    Intracellular products, not secreted from the microbial cell, are released by breaking the cell envelope consisting of cytoplasmic membrane and an outer cell wall. Hydrodynamic cavitation has been reported to cause microbial cell disruption. By manipulating the operating variables involved, a wide range of intensity of cavitation can be achieved resulting in a varying extent of disruption. The effect of the process variables including cavitation number, initial cell concentration of the suspension and the number of passes across the cavitation zone on the release of enzymes from various locations of the Brewers' yeast was studied. The release profile of the enzymes studied include alpha-glucosidase (periplasmic), invertase (cell wall bound), alcohol dehydrogenase (ADH; cytoplasmic) and glucose-6-phosphate dehydrogenase (G6PDH; cytoplasmic). An optimum cavitation number Cv of 0.13 for maximum disruption was observed across the range Cv 0.09-0.99. The optimum cell concentration was found to be 0.5% (w/v, wet wt) when varying over the range 0.1%-5%. The sustained effect of cavitation on the yeast cell wall when re-circulating the suspension across the cavitation zone was found to release the cell wall bound enzyme invertase (86%) to a greater extent than the enzymes from other locations of the cell (e.g. periplasmic alpha-glucosidase at 17%). Localised damage to the cell wall could be observed using transmission electron microscopy (TEM) of cells subjected to less intense cavitation conditions. Absence of the release of cytoplasmic enzymes to a significant extent, absence of micronisation as observed by TEM and presence of a lower number of proteins bands in the culture supernatant on SDS-PAGE analysis following hydrodynamic cavitation compared to disruption by high-pressure homogenisation confirmed the selective release offered by hydrodynamic cavitation.

  12. Evaluation of indigenous grains from the Peruvian Andean region for antidiabetes and antihypertension potential using in vitro methods.

    Science.gov (United States)

    Ranilla, Lena Galvez; Apostolidis, Emmanouil; Genovese, Maria Ines; Lajolo, Franco Maria; Shetty, Kalidas

    2009-08-01

    The health-relevant functionality of 10 thermally processed Peruvian Andean grains (five cereals, three pseudocereals, and two legumes) was evaluated for potential type 2 diabetes-relevant antihyperglycemia and antihypertension activity using in vitro enzyme assays. Inhibition of enzymes relevant for managing early stages of type 2 diabetes such as hyperglycemia-relevant alpha-glucosidase and alpha-amylase and hypertension-relevant angiotensin I-converting enzyme (ACE) were assayed along with the total phenolic content, phenolic profiles, and antioxidant activity based on the 1,1-diphenyl-2-picrylhydrazyl radical assay. Purple corn (Zea mays L.) (cereal) exhibited high free radical scavenging-linked antioxidant activity (77%) and had the highest total phenolic content (8 +/- 1 mg of gallic acid equivalents/g of sample weight) and alpha-glucosidase inhibitory activity (51% at 5 mg of sample weight). The major phenolic compound in this cereal was protocatechuic acid (287 +/- 15 microg/g of sample weight). Pseudocereals such as Quinoa (Chenopodium quinoa Willd) and Kañiwa (Chenopodium pallidicaule Aellen) were rich in quercetin derivatives (1,131 +/- 56 and 943 +/- 35 microg [expressed as quercetin aglycone]/g of sample weight, respectively) and had the highest antioxidant activity (86% and 75%, respectively). Andean legumes (Lupinus mutabilis cultivars SLP-1 and H-6) inhibited significantly the hypertension-relevant ACE (52% at 5 mg of sample weight). No alpha-amylase inhibitory activity was found in any of the evaluated Andean grains. This in vitro study indicates the potential of combination of Andean whole grain cereals, pseudocereals, and legumes to develop effective dietary strategies for managing type 2 diabetes and associated hypertension and provides the rationale for animal and clinical studies.

  13. New treatments for patients with type 2 diabetes mellitus.

    Science.gov (United States)

    Wolffenbuttel, B H; Graal, M B

    1996-11-01

    In subjects with type 2 diabetes, both defects of insulin secretion and insulin resistance contribute to the development of hyperglycaemia. The major goals of treatment are to optimise blood glucose control, and normalise the associated lipid disturbances and elevated blood pressure. Pharmacologic treatment is often necessary. This paper discusses new forms of oral treatment for subjects with type 2 diabetes. These include a new sulphonylurea compound glimepiride (Amaryl), which binds to a different protein of the putative sulphonylurea receptor than glibenclamide, and seems to have a lower risk of hypoglycaemia. A new class of drugs with insulin secretory capacity, of which repaglinide (NovoNorm) is the leading compound, is now in phase III clinical trials. Alpha-glucosidase inhibitors reversibly inhibit alpha-glucosidase enzymes in the small intestine, which delays cleavage of oligo- and disaccharides to monosaccharides. This leads to a delayed and reduced blood glucose rise after a meal. Two compounds are in development or have been marketed, ie, miglitol and acarbose (Glucobay). Another new class of drugs is the thiazolidine-diones, which seem to work by enhancing insulin action. The 'insulin sensitising' effects of the leading compounds, troglitazone and BRL 49653C, do not involve any effect on insulin secretion. These drugs also seem to beneficially influence serum cholesterol and triglyceride levels. Oral antihyperglycaemic agents can be used only during a limited period of time in most patients, after which the diabetic state 'worsens' and insulin therapy has to be started. In this light, two new forms of treatment which require subcutaneous injections are also discussed: the synthetic human amylin analogue AC137 (pramlintide) and glucagon-like peptide-1 (7-36)-amide, a strong glucose-dependent stimulator of insulin secretion. It remains to be seen whether these compounds can be developed further for clinical use in patients with diabetes.

  14. Prunella vulgaris L. active components and their hypoglycemic and antinociceptive effects in alloxan-induced diabetic mice.

    Science.gov (United States)

    Raafat, K; Wurglics, M; Schubert-Zsilavecz, M

    2016-12-01

    Prunella vulgaris L. (Lamiaceae) (PV) is a herbaceous plant traditionally utilized in management of diabetes and it has immunomodulatory activity. In this study, acute and subchronic antidiabetic, in-vivo antioxidant and antinociceptive effects of PV were evaluated in alloxan-induced type 1 diabetes (T1D) in a mouse model. Bio-guided fractionation, isolation, RP-HPLC, and (1)H and (13)C NMR identification of the active components responsible for PV effects were determined. RP-HPLC analysis showed that PV contained rosmarinic acid (RA) 4.5%, caffeic acid (CA) 9.8% and p-coumaric acid (pCA) 11.6%. Bio-guided fractionation showed that PV most active fraction was rich in caffeic acid, hence named, caffeic acid-rich fraction (CARF). RP-HPLC, and (1)H and (13)C NMR experiments showed that CARF contained CA (93.4%) and RA (6.6%). CARF reduced blood glucose levels and improved in-vivo oxidative-stress. It also inhibited the carbohydrate-hydrolyzing enzymes (alpha-amylase and alpha-glucosidase) and reduced HbA1c levels more significantly (p≤0.05) than that of PV and equivalent amounts of CA or RA. For longer times, CARF had significantly (p≤0.05) increased serum-insulin, ameliorated thermal hyperalgesia and tactile allodynia more significantly (p≤0.05) than the effects of PV and equivalent amounts of CA or RA. Moreover, the tested compounds showed potential restoration of the lipid peroxide levels. Consequently, CARF and PV observed increase in serum-insulin, attenuation of alpha-amylase and alpha-glucosidase, and their antioxidant potentials might be responsible for their antidiabetogenic and antinociceptive properties. In conclusion, CARF isolated from PV could be a potential therapeutic agent to ameliorate T1D and related complications.

  15. A new diagnostic assay for glycogen storage disease type II in mixed leukocytes.

    Science.gov (United States)

    Okumiya, Toshika; Keulemans, Joke L M; Kroos, Marian A; Van der Beek, Nadine M E; Boer, Marijke A; Takeuchi, Hiroaki; Van Diggelen, Otto P; Reuser, Arnold J J

    2006-05-01

    We have established a new method for the enzymatic diagnosis of glycogen storage disease type II (Pompe disease or acid maltase deficiency) using mixed leukocytes. The method employs glycogen and 4-methylumbelliferyl-alpha-D-glucopyranoside (4MU-alphaGlc) as substrates for measuring the lysosomal acid alpha-glucosidase (acid alphaGlu) activity, and incorporates acarbose to eliminate the interference of unrelated alpha-glucosidases (predominantly maltase-glucoamylase). It is shown that 3.0 micromol/L acarbose completely inhibits the maltase-glucoamylase activity at pH 4.0, but the lysosomal acid alphaGlu activity by less than 5%. With this method, we determined the acid alphaGlu activity in mixed leukocytes from 25 patients with glycogen storage disease type II (2 infantile and 23 late-onset cases), one GAA2/GAA2 homozygote and 30 healthy subjects. In the assay with glycogen as substrate, the addition of acarbose created a clear separation between the patient and the control ranges. In the assay with 4MU-alphaGlc as substrate, the two ranges were fully separated but remained very close despite the use of acarbose. The separation of the patient and normal ranges was improved considerably by taking the ratio of acarbose-inhibited over uninhibited activity. A GAA2/GAA2 homozygote was correctly diagnosed with 4MU-alphaGlc but misdiagnosed as patient when glycogen was used as substrate. We conclude that the inclusion of 3.0 micromol/L acarbose in the assays with glycogen and 4MU-alphaGlc substrates at pH 4.0 allows for the specific measurement of lysosomal acid alphaGlu activity in mixed leukocytes, thus enabling a reliable diagnosis of glycogen storage disease type II in this specimen.

  16. Effect of experimental varicocele on structure and function of epididymis in adolescent rats

    Institute of Scientific and Technical Information of China (English)

    Qiu-YangZHANG; Shu-DongQIU; Xiao-NianMA; He-MingYU; Yan-WanWU

    2003-01-01

    Aim: To study the effect of experimental left varicocele (ELV) on epididymal structure and function in adolescent Sprague-Dawley rats. Methods: ELV was induced by partial ligation of the left renal vein. Sham-operated animals served as the controls. Four and 8 weeks after the operation, the histological, ultrastructural and biochemical(alpha-glucosidase activity and camitine content) changes in different segments of the epididymis were observed.Results: In the treated animals, there were degeneration of the epididymal epithelium and edema of the interstitial tissue; numerous shedding cells, residual bodies, deformed sperm and macrophages appeared in the epididymal lumen.Morphometric measurement indicated a significant reduction in the epididymal tubular diameter (P<0.05) and a significant increase in the epididymal interstitial area (P<0.05) compared with the controls. Ultrastructural study showed sparse microvilli of the columnar epithelium, increased and enlarged lysosomes in the principal cells with defected organelles and the presence of large cytoplasmic vacuoles. The protein and carnitine contents and the alpha-glucosi-dase activity in the caput, corpus and cauda epididymis of the ELV rats were lower than those of the controls (P<0.05). Conclusion: There were structural and functional changes in the epididymis of adolescent ELV rats, which may contribute to the infertility caused by varicocele. (Asian J Androl 2003 Jun; 5: 108-112)

  17. Microbial dextran-hydrolyzing enzymes: fundamentals and applications.

    Science.gov (United States)

    Khalikova, Elvira; Susi, Petri; Korpela, Timo

    2005-06-01

    Dextran is a chemically and physically complex polymer, breakdown of which is carried out by a variety of endo- and exodextranases. Enzymes in many groups can be classified as dextranases according to function: such enzymes include dextranhydrolases, glucodextranases, exoisomaltohydrolases, exoisomaltotriohydrases, and branched-dextran exo-1,2-alpha-glucosidases. Cycloisomalto-oligosaccharide glucanotransferase does not formally belong to the dextranases even though its side reaction produces hydrolyzed dextrans. A new classification system for glycosylhydrolases and glycosyltransferases, which is based on amino acid sequence similarities, divides the dextranases into five families. However, this classification is still incomplete since sequence information is missing for many of the enzymes that have been biochemically characterized as dextranases. Dextran-degrading enzymes have been isolated from a wide range of microorganisms. The major characteristics of these enzymes, the methods for analyzing their activities and biological roles, analysis of primary sequence data, and three-dimensional structures of dextranases have been dealt with in this review. Dextranases are promising for future use in various scientific and biotechnological applications.

  18. The Active Role of Leguminous Plant Components in Type 2 Diabetes

    Directory of Open Access Journals (Sweden)

    Monika Gętek

    2014-01-01

    Full Text Available Diabetes appears to be one of the most frequent noncommunicable diseases in the world. A permanent growth in the incidence of diabetes can be observed and according to the International Diabetes Federation (IDF the year 2030 will mark the increase in the number of diabetics to 439 mln worldwide. Type 2 diabetes accounts for about 90% of all diabetes incidence. Nutrition model modification not only features the basic element in type 2 diabetes treatment but also constitutes the fundamental factor influencing a morbidity rate decrease. Leguminous plants are a key factor in the diabetic diet; plants such as pulses or soybeans are nutritious products valued highly in nutrition. These legumes are high in the content of wholesome protein and contain large amounts of soluble alimentary fiber fractions, polyunsaturated fatty acids, vitamins and minerals, and bioactive substances with antioxidant, anti-inflammatory, and anticancer activity. They are distinguished by the high amount of bioactive compounds that may interfere with the metabolism of glucose. The most significant bioactive compounds displaying antidiabetic activity in leguminous plants are as follows: genistein and daidzein, alpha-amylase inhibitors, and alpha-glucosidase inhibitors. In vitro research using leguminous plant extracts has confirmed their antidiabetic properties. Leguminous plants should be employed in the promotion of healthy lifestyles in terms of functional food.

  19. Evaluation of the toxic effects of arsenite, chromate, cadmium, and copper using a battery of four bioassays

    Energy Technology Data Exchange (ETDEWEB)

    Ko, Kyung-Seok; Lee, Pyeong-Koo [Korea Institute of Geoscience and Mineral Resources (KIGAM), Daejeon (Korea, Republic of). Geologic Environment Div.; Kong, In Chul [Yeungnam Univ., Kyungbuk (Korea, Republic of). Dept. of Environmental Engineering

    2012-09-15

    The sensitivities of four different kinds of bioassays to the toxicities of arsenite, chromate, cadmium, and copper were compared. The different bioassays exhibited different sensitivities, i.e., they responded to different levels of toxicity of each of the different metals. However, with the exception of the {alpha}-glucosidase enzyme activity, arsenite was the most toxic compound towards all the tested organisms, exhibiting the highest toxic effect on the seeds of Lactuca, with an EC{sub 50} value of 0.63 mg/L. The sensitivities of Lactuca and Raphanus were greater than the sensitivities of two other kinds of seeds tested. Therefore, these were the seeds appropriate for use in a seed germination assay. A high revertant mutagenic ratio (5:1) of Salmonella typhimurium was observed with an arsenite concentration of 0.1 {mu}g/plate, indicative of a high possibility of mutagenicity. These different results suggested that a battery of bioassays, rather than one bioassay alone, is needed as a more accurate and better tool for the bioassessment of environmental pollutants. (orig.)

  20. Impact of infection on the secretory capacity of the male accessory glands

    Directory of Open Access Journals (Sweden)

    M. Marconi

    2009-06-01

    Full Text Available INTRODUCTION: Studies that compare the impact of different infectious entities of the male reproductive tract (MRT on the male accessory gland function are controversial. MATERIAL AND METHODS: Semen analyses of 71 patients with proven infections of the MRT were compared with the results of 40 healthy non-infected volunteers. Patients were divided into 3 groups according to their diagnosis: chronic prostatitis NIH type II (n = 38, chronic epididymitis (n = 12, and chronic urethritis (n = 21. RESULTS: The bacteriological analysis revealed 9 different types of microorganisms, considered to be the etiological agents, isolated in different secretions, including: urine, expressed prostatic secretions, semen and urethral smears: E. Coli (n = 20, Klebsiella (n = 2, Proteus spp. (n = 1, Enterococcus (n = 20, Staphylococcus spp. (n = 1, M. tuberculosis (n = 2, N. gonorrhea (n = 8, Chlamydia tr. (n = 16 and, Ureaplasma urealyticum (n = 1. The infection group had significantly (p < 0.05 lower: semen volume, alpha-glucosidase, fructose, and zinc in seminal plasma and, higher pH than the control group. None of these parameters was sufficiently accurate in the ROC analysis to discriminate between infected and non-infected men. CONCLUSION: Proven bacterial infections of the MRT impact negatively on all the accessory gland function parameters evaluated in semen, suggesting impairment of the secretory capacity of the epididymis, seminal vesicles and prostate. These findings were associated with an infectious related significant increase of semen pH. None of the semen parameters evaluated can be suggested as a diagnostic tool for infection.

  1. A quick overview on some aspects of endocrinological and therapeutic effects of Berberis vulgaris L.

    Science.gov (United States)

    Zarei, Ali; Changizi-Ashtiyani, Saeed; Taheri, Soheila; Ramezani, Majid

    2015-01-01

    Many herbaceous plants contain compounds that have biological effects in addition to their medicinal properties. They have compounds with numerous properties, including hypo lipidemic, hypoglycemic, antioxidant, and hepato protective ones, which have been analyzed at different levels. One of these plants, with the scientific name of Berberis vulgaris, is barberry. The most important compounds identified in this plant are berberine, oxycontin, palmatine, bervulcine, berbamine, columbamine, jatrorrhizine, coptisine, and berbamine. In addition to alkaloids, organic acids such as chelidonic acid, citric acid, malic acid, resin, tannin, pectinic, and mucilagic substances are among the ingredients of barberry. In this paper, it was attempted to determine the role and effect of the extract of barberry on various body organs. The results showed that berberine actually increases insulin sensitivity and is capable of inhibiting alpha glucosidase, adipogenesis, and thus acts as an anti-obesity and hypoglycemic agent. Berberine reduces the density of serum cholesterol and triglycerides and can improve the function of liver enzymes, therefore, it can be suggested as a hypo lipidemic and hepato protective plant extract. The hepato protective effects of this extract are probably due to its antioxidant properties. Studies showed that barberry have numerous health benefits, including anti-inflammatory ones. Moreover, it can be used as a medicinal herb to treat a variety of disorders, such as diabetes, liver disease, gallbladder pain, digestive, urinary tract diseases, and gallstones. However, more studies on this issue and doing more focused and intensive researches in this field are recommended.

  2. Microbial amylolytic enzymes.

    Science.gov (United States)

    Vihinen, M; Mäntsälä, P

    1989-01-01

    Starch-degrading, amylolytic enzymes are widely distributed among microbes. Several activities are required to hydrolyze starch to its glucose units. These enzymes include alpha-amylase, beta-amylase, glucoamylase, alpha-glucosidase, pullulan-degrading enzymes, exoacting enzymes yielding alpha-type endproducts, and cyclodextrin glycosyltransferase. Properties of these enzymes vary and are somewhat linked to the environmental circumstances of the producing organisms. Features of the enzymes, their action patterns, physicochemical properties, occurrence, genetics, and results obtained from cloning of the genes are described. Among all the amylolytic enzymes, the genetics of alpha-amylase in Bacillus subtilis are best known. Alpha-Amylase production in B. subtilis is regulated by several genetic elements, many of which have synergistic effects. Genes encoding enzymes from all the amylolytic enzyme groups dealt with here have been cloned, and the sequences have been found to contain some highly conserved regions thought to be essential for their action and/or structure. Glucoamylase appears usually in several forms, which seem to be the results of a variety of mechanisms, including heterogeneous glycosylation, limited proteolysis, multiple modes of mRNA splicing, and the presence of several structural genes.

  3. Purification, biochemical characterization, and gene cloning of a new extracellular thermotolerant and glucose tolerant maltooligosaccharide-forming alpha-amylase from an endophytic ascomycete Fusicoccum sp. BCC4124.

    Science.gov (United States)

    Champreda, Verawat; Kanokratana, Pattanop; Sriprang, Rutchadaporn; Tanapongpipat, Sutipa; Eurwilaichitr, Lily

    2007-08-01

    An endophytic fungus, Fusicoccum sp. BCC4124, showed strong amylolytic activity when cultivated on multi-enzyme induction enriched medium and agro-industry substrates. alpha-Amylase and alpha-glucosidase activities were highly induced in the presence of maltose and starch. The purified target alpha-amylase, Amy-FC1, showed strong hydrolytic activity on soluble starch (kcat/Km=6.47 x 10(3) min(-1)(ml/mg)) and selective activity on gamma- and beta-cyclodextrins, but not on alpha-cyclodextrin. The enzyme worked optimally at 70 degrees C in a neutral pH range with t(1/2) of 240 min in the presence of Ca(2+) and starch. Maltose, matotriose, and maltotetraose were the major products from starch hydrolysis but prolonged reaction led to the production of glucose, maltose, and maltotriose from starch, cyclodextrins, and maltooligosaccharides (G3-G7). The amylase showed remarkable glucose tolerance up to 1 M, but was more sensitive to inhibition by maltose. The deduced protein primary structure from the putative gene revealed that the enzyme shared moderate homology between alpha-amylases from Aspergilli and Lipomyces sp. This thermotolerant, glucose tolerant maltooligosaccharide-forming alpha-amylase is potent for biotechnological application.

  4. The microbial production of amylase inhibitor and its application. I. Isolation and cultivation of Streptomyces nigrifaciens NTU-3314.

    Science.gov (United States)

    Su, Y C; Chiu, R J; Yu, N; Chang, W R

    1984-10-01

    In the course of screening amylase inhibitor producing, microorganisms, a strain identified as Streptomyces nigrifaciens NTU-3314 was found to have the highest inhibitor-producing ability among the other isolated strains. This strain was aerobically cultured at 30 degrees C in a 5l jar fermentor with a working volume of 2l. The optimum cultural medium consisted of defatted soybean flake 3.0%, potato starch 4.0%, casein 0.6%, sucrose 0.6%, serine 0.02% and NaCl 0.8% (pH 7.0). With an aeration rate of 1.5 vvm, an agitation speed of 300 rpm and an inoculum of 15% seed (previously grown in seed medium 3), the highest amount of inhibitor was obtained after 24 hours of cultivation. The amylase inhibitor produced had inhibitory effects on both alpha-amylase and glucoamylase, but not on beta-amylase, alpha-glucosidase, beta-glucosidase or dextranase. It was quite stable in 0.1M phosphate buffer (pH 7.0) and nearly 100% of its activity was retained even after boiling at 100 degrees C for 20 min.

  5. [Synthesis of novel beta-aminoalcohols containing nabumetone moiety with potential antidiabetic activity].

    Science.gov (United States)

    Zhang, Kun; Yan, Ju-fang; Tang, Xue-mei; Liu, Hong-ping; Fan, Li; Zhou, Guang-ming; Yang, Da-cheng

    2011-04-01

    Twenty five new beta-aminoalcohols containing nabumetone moiety were prepared via the reduction of potassium borohydride with a convenient and efficient procedure, starting from beta-aminoketones that have been synthesized by our group. Their chemical structures were determined by IR, MS, 1H NMR, 13C NMR, HR-MS and antidiabetic activities were screened in vitro. Preliminary results revealed that the antidiabetic activity of most beta-aminoalcohols were better than that of the corresponding beta-aminoketones. Although most compounds showed weak antidiabetic activity, the alpha-glucosidase inhibitory activity of compounds 5hd(1) and 5id(2) reached 74.37% and 90.15%, respectively, which were superior to the positive control. The relative peroxisome proliferator-activated receptor response element (PPRE) activity of five compounds were more than 60%, among them compound 5ca possessed the highest activity (112.59%). As lead molecules of antidiabetic agents, compounds 5hd(1), 5id(2) and 5ca deserve further study.

  6. Successful pregnancy and birth after intrauterine insemination using caput epididymal sperm by percutaneous aspiration

    Institute of Scientific and Technical Information of China (English)

    YiQIU; Dan-TongYANG; Su-MeiWANG; Hui-QingSUN; Yi-FangJIA

    2003-01-01

    Aim:To manage male infertility with obstructive azoospermia by means of percutaneous epididymal sperm aspiration(PESA)and intrauterine insemination(IUI),Methods:Ninety azoospermic patients with congenital bilateral absence of the vas deferens(BAVD,N=58)or bilateral caudal epididymal obstruction(BCEO,n=32)requesting for fine needle aspiration(FNA),PESA and IUI were recruited.The obstruction was diagnosed by vasography and determination of the fructose,carnitine and alpha-glucosidase levels in the seminal fluid.Results:The mean sperm motility,density,abnormal sperm and total sperm count of the caput epdidymis were 16%±22%,(12±31)×106/mL,55%±36% and (16±14)×106,respectively,In the 90 couples,a total of 74 PESA procedures and 66 cycles of IUI were performed.Three pregnancies resulted,including one twin pregnancy giving birth to two healthy boys, one single pregnancy with a healthy girl and another single pregnancy aborted at week 6 of conception.The pregnancy rate per IUI cycle was 4.5%.Conclusiong:The birth of normal,healthy infants by IUI using PESA indicates that the caput epididymal sperm possess fertilization capacity.The PESA-IUI programme is a practical and economical procedure for the management of patients with obstructive azoospermia.

  7. Anti-hyperglycemic and anti-hyperlipidemic effects of guava leaf extract

    Science.gov (United States)

    2010-01-01

    Psidium guajava Linn. (guava) is used not only as food but also as folk medicine in subtropical areas around the world because of its pharmacologic activities. In particular, the leaf extract of guava has traditionally been used for the treatment of diabetes in East Asia and other countries. Moreover, the anti-hyperglycemic activity of the extract has been reported in some animal models. However, little is known regarding the therapeutic activity of the extract in human clinical trials as well as its underlying therapeutic mechanisms and safety. In Japan, Guava Leaf Tea (Bansoureicha®, Yakult Honsha, Tokyo, Japan) containing the aqueous leaf extract from guava has been approved as one of the Foods for Specified Health Uses and is now commercially available. This review describes the active component of the aqueous guava leaf extract and its inhibition of alpha-glucosidase enzymes in vitro, safety of the extract and Guava Leaf Tea, reduction of postprandial blood glucose elevation, and improvement of hyperglycemia, hyperinsulinemia, hypoadiponectinemia, hypertriglycemia and hypercholesterolemia in murine models and several clinical trials. It is suggested that the chronic suppression of postprandial blood glucose elevation is important in preventing type 2 diabetes mellitus, and that Guava Leaf Tea is considered useful as an alimentotherapy for chronic treatment. PMID:20181067

  8. Effect of pioglitazone on muscle sympathetic nerve activity in type 2 diabetes mellitus with α-glucosidase inhibitor.

    Science.gov (United States)

    Kobayashi, Daisuke; Takamura, Masayuki; Murai, Hisayoshi; Usui, Soichiro; Ikeda, Tatsunori; Inomata, Jun-ichiro; Takashima, Shin-ichiro; Kato, Takeshi; Furusho, Hiroshi; Takeshita, Yumie; Ota, Tsuguhito; Takamura, Toshinari; Kaneko, Shuichi

    2010-12-08

    Activation of the sympathetic nervous system is augmented in patients with type 2 diabetes mellitus (DM). Pioglitazone, an anti-diabetic drug, improves insulin resistance, but its influence on sympathetic nerve activity is not clear. To identify the relationship between insulin resistance and sympathetic activity, we examined muscle sympathetic nerve activity (MSNA) in controlled type 2 DM patients with alpha-glucosidase inhibitor (GI). We measured MSNA and calculated homeostasis model assessment of insulin resistance index (HOMA-IR) in twelve DM patients treated with alpha-GI and thirteen age-matched healthy subjects. In DM patients with alpha-GI, all parameters were reexamined after three months of treatment with pioglitazone. MSNA and HOMA-IR were significantly greater in DM patients with alpha-GI compared to healthy subjects. Hemoglobin A1c did not differ in DM patients before and after pioglitazone. However, pioglitazone significantly decreased MSNA in DM patients compared with alpha-GI (21.7±5.2 vs. 32.0±6.8 burst/min, ppioglitazone was similar to that in healthy subjects. HOMA-IR significantly decreased after pioglitazone, and a significant relationship was found between the absolute change in MSNA and HOMA-IR (r=0.65, ppioglitazone provides an additional effect on inhibition of sympathetic nerve activity.

  9. Preventive pharmacotherapy in type 2 diabetes mellitus.

    Science.gov (United States)

    Choudhary, Neeraj; Kalra, Sanjay; Unnikrishnan, Ambika Gopalkrishnan; Ajish, T P

    2012-01-01

    Over the last few decades certain demographic changes have been observed worldwide, which have led to an increase in the prevalence of chronic non-communicable diseases. Type 2 diabetes mellitus and associated cardiovascular disease are major contributors to this disease burden leading to rising morbidity and mortality. It is worrisome to see that type 2 diabetes with its micro- and macrovascular complications is occurring in younger populations where it was hitherto unseen. Prevention appears to be an important strategy to reduce the burden of disease. Along with inculcating healthy lifestyle habits across populations, it may be suitable to use preventive pharmacotherapy in those with pre-diabetes and / or other risk factors like obesity, hypertension, and on the like. Metformin, alpha glucosidase inhibitors like acarbose, miglitol, and voglibose, and pioglitazone have all been used with success. The issues of compliance and adverse effects during long-term use have tempered the use of these drugs. The best approach would be to motivate the patient for effective lifestyle changes, and pharmacological management if the lifestyle changes are not successful in achieving their goals.

  10. Diabetes mellitus and kidney disease in the elderly.

    Science.gov (United States)

    Iglesias, Pedro; Heras, Manuel; Díez, Juan J

    2014-05-21

    Management of diabetic elderly patients with chronic kidney disease involves specific characteristics that affect both metabolic control and therapeutic measures. Blood glucose control targets should be individualised based on life expectancy, renal function, hypoglycaemia risk and comorbidity. Metformin may be used alone or in combination with other oral anti-diabetic drugs but must be discontinued when the glomerular filtration rate is less than 30 mL/min. Gliclazide and glipizide are sulfonylureas that do not require dose adjustment in chronic kidney disease but they should be avoided in cases of advanced kidney disease because of the risk of hypoglycaemia. Repaglinide is the only meglitinide recommended in these patients. Alpha-glucosidase inhibitors must be avoided in patients with a glomerular filtration rate of less than 25 mL/min or those undergoing dialysis. Pioglitazone does not require dose adjustment but it has potentially adverse effects in this population. Dipeptidyl peptidase-4 inhibitors are effective and well tolerated. Of the latter, linagliptin does not require dose adjustment. Glucagon-like peptide-1 receptor agonists and sodium-glucose cotransporter 2 inhibitors are not recommended in elderly patients with advanced kidney disease. Lastly, insulin therapy, particularly using the new insulin analogues, allows adequate management of hyperglycaemia in these patients, with different therapeutic regimens that must be individualised in order to avoid hypoglycaemia.

  11. Effervescent Granules Prepared Using Eucommia ulmoides Oliv. and Moso Bamboo Leaves: Hypoglycemic Activity in HepG2 Cells

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    Xiang-Zhou Li

    2016-01-01

    Full Text Available Eucommia ulmoides Oliv. (E. ulmoides Oliv. and moso bamboo (Phyllostachys pubescens leaves are used as folk medicines in central-western China to treat diabetes. To investigate the hypoglycemic activity of the effervescent granules prepared using E. ulmoides Oliv. and moso bamboo leaves (EBEG in HepG2 cells, EBEG were prepared with 5% of each of polysaccharides and chlorogenic acids from moso bamboo and E. ulmoides Oliv. leaves, respectively. HepG2 cells cultured in a high-glucose medium were classified into different groups. The results displayed EBEG-treated cells showed better glucose utilization than the negative controls; thus, the hypoglycemic effect of EBEG was much greater than that of granules prepared using either component alone, thereby indicating that this effect was due to a synergistic action of the components. Further, glucose consumption levels in the cells treated with EBEG (156.35% at 200 μg/mL and the positive controls (metformin, 162.29%; insulin, 161.52% were similar. Thus, EBEG exhibited good potential for use as a natural antidiabetic agent. The hypoglycemic effect of EBEG could be due to the synergistic action of polysaccharides from the moso bamboo leaves and chlorogenic acids from E. ulmoides Oliv. leaves via the inhibition of alpha-glucosidase and glucose-6-phosphate displacement enzyme.

  12. Action pattern of human pancreatic and salivary alpha-amylase on 1,4-alpha-D-nitrophenylmaltooligosaccharides. 1,4-alpha-D-nitrophenylmaltooligosaccharides as substrates of alpha-amylse, I.

    Science.gov (United States)

    Wallenfels, K; Laule, G; Meltzer, B

    1982-08-01

    High performance liquid chromatography (HPLC) was used to monitor the purity of the substrates and to establish the patterns of hydrolysis of ortho- and para-nitrophenylmaltooligosaccharides (2-7 glucose residues) catalysed by human pancreatic and salivary alpha-amylase. Separation of the reaction products from the remaining substrate was performed on a TSK-G-2000 PW or a RP18 column. By measuring the quantitative distribution of products, and assuming a 5-subsite model for the active site of alpha-amylase, differential activities for the hydrolysis of the different glycosidic bonds in the 2 series of substrates were deduced. A highly sensitive coupled continuous assay system is based on the formation of phenyloligosaccharides with 1-4 glucose residues by the action of the amylase under test, coupled to hydrolysis of these products by yeast alpha-glucosidase. The most suitable test substrates were shown to be para-nitrophenyl-alpha-D-maltotetraoside and -pentaoside. Direct production of nitrophenol from ortho-nitrophenyl-alpha-D-maltotrioside is recommended for the measurement of the alpha-amylase activity of pancreatic and salivary gland secretions and extracts.

  13. Phenolic compounds, antioxidant activity and in vitro inhibitory potential against key enzymes relevant for hyperglycemia and hypertension of commonly used medicinal plants, herbs and spices in Latin America.

    Science.gov (United States)

    Ranilla, Lena Galvez; Kwon, Young-In; Apostolidis, Emmanouil; Shetty, Kalidas

    2010-06-01

    Traditionally used medicinal plants, herbs and spices in Latin America were investigated to determine their phenolic profiles, antioxidant activity and in vitro inhibitory potential against key enzymes relevant for hyperglycemia and hypertension. High phenolic and antioxidant activity-containing medicinal plants and spices such as Chancapiedra (Phyllantus niruri L.), Zarzaparrilla (Smilax officinalis), Yerba Mate (Ilex paraguayensis St-Hil), and Huacatay (Tagetes minuta) had the highest anti-hyperglycemia relevant in vitro alpha-glucosidase inhibitory activities with no effect on alpha-amylase. Molle (Schinus molle), Maca (Lepidium meyenii Walp), Caigua (Cyclanthera pedata) and ginger (Zingiber officinale) inhibited significantly the hypertension relevant angiotensin I-converting enzyme (ACE). All evaluated pepper (Capsicum) genus exhibited both anti-hyperglycemia and anti-hypertension potential. Major phenolic compounds in Matico (Piper angustifolium R.), Guascas (Galinsoga parviflora) and Huacatay were chlorogenic acid and hydroxycinnamic acid derivatives. Therefore, specific medicinal plants, herbs and spices from Latin America have potential for hyperglycemia and hypertension prevention associated with Type 2 diabetes.

  14. Natural products as promising drug candidates for the treatment of hepatitis B and C.

    Science.gov (United States)

    Wohlfarth, Carolin; Efferth, Thomas

    2009-01-01

    Hepatitis B virus (HBV) or hepatitis C virus (HCV) infections are a major threat worldwide. Combination therapy of interferon-alpha and ribavirin is currently the treatment of choice for HCV-infected patients. However, this regimen is only effective in approximately 50% of patients and provokes severe side-effects. Numerous natural alternatives for treating HCV have been suggested. Deoxynojirimycin and its derivatives are iminosugars which exert anti-HCV activity by inhibiting alpha-glucosidases. A non-immunosuppressive derivate of cyclosporine A, NIM811, exerts anti-HCV activity by binding to cyclophilin. Other natural products with promising anti-HCV activity are 2-arylbenzofuran derivatives, Mellein, and pseudoguaianolides. For HBV treatment, several drugs are available, specifically targeting the virus polymerase (lamivudine, entecavir, telbivudine, and adefovir dipivoxil). The efficacy of these drugs is hampered by the development of resistance due to point mutations in the HBV polymerase. Due to drug resistance and adverse side-effects, the search for novel drugs is mandatory. Wogonin, ellagic acid, artemisinin and artesunate, chrysophanol 8-O-beta-D-glucoside, saikosaponin C, and protostane triterpenes are active against HBV. Natural products need to be investigated in more detail to explore their potential as novel adjuncts to established HBV or HCV therapy.

  15. Discovery of insect and human dengue virus host factors.

    Science.gov (United States)

    Sessions, October M; Barrows, Nicholas J; Souza-Neto, Jayme A; Robinson, Timothy J; Hershey, Christine L; Rodgers, Mary A; Ramirez, Jose L; Dimopoulos, George; Yang, Priscilla L; Pearson, James L; Garcia-Blanco, Mariano A

    2009-04-23

    Dengue fever is the most frequent arthropod-borne viral disease of humans, with almost half of the world's population at risk of infection. The high prevalence, lack of an effective vaccine, and absence of specific treatment conspire to make dengue fever a global public health threat. Given their compact genomes, dengue viruses (DENV-1-4) and other flaviviruses probably require an extensive number of host factors; however, only a limited number of human, and an even smaller number of insect host factors, have been identified. Here we identify insect host factors required for DENV-2 propagation, by carrying out a genome-wide RNA interference screen in Drosophila melanogaster cells using a well-established 22,632 double-stranded RNA library. This screen identified 116 candidate dengue virus host factors (DVHFs). Although some were previously associated with flaviviruses (for example, V-ATPases and alpha-glucosidases), most of the DVHFs were newly implicated in dengue virus propagation. The dipteran DVHFs had 82 readily recognizable human homologues and, using a targeted short-interfering-RNA screen, we showed that 42 of these are human DVHFs. This indicates notable conservation of required factors between dipteran and human hosts. This work suggests new approaches to control infection in the insect vector and the mammalian host.

  16. Conversion of cassava starch to biomass, carbohydrates, and acids by Aspergillus niger.

    Science.gov (United States)

    Tan, K H; Ferguson, L B; Carlton, C

    1984-01-01

    The filamentous fungus, Aspergillus niger, efficiently converted cassava polysaccharides to mycelial mass, simple sugars, and acids during the course of its growth. A typical 70-ml culture broth containing 2% cassava polysaccharides yielded 0.38 g dry mycelial mass, 1.14 mmol reducing sugars, and 1.17 meq acids at the end of 42 h. About 70% of the initial total carbohydrate in the medium was degraded during the same period. The sugars and acids in the culture broths were analyzed by HPLC on a single Aminex HPX-87 column at 55 degrees C, using 0.013 N H2SO4 as the eluting solvent. Cassava polysaccharides were degraded to oligosaccharides, maltotriose, maltose, and glucose beyond the 20-h growth periods, with maltotriose emerging as the major simple sugar. The appearance of citric, malic, gluconic, succinic, and fumaric acids accounted mostly for the decreasing pH in the growth media. Formation of carbohydrate species in the culture broths was closely related to the biosynthesis and secretion of several carbohydrases by A. niger. The extracellular carbohydrases were separated and identified by chromatofocusing and polyacrylamide gel electrophoresis to be amyloglucosidase (EC 3.1.2.3), alpha-amylase (EC 3.2.1.1), and alpha-glucosidase (EC 3.2.1.20), respectively.

  17. Comparison of media for the isolation of Enterobacter sakazakii.

    Science.gov (United States)

    Iversen, Carol; Forsythe, Stephen J

    2007-01-01

    Enterobacter sakazakii is associated with neonatal infections and is occasionally present at low levels (ESSB), and modified lauryl sulfate broth (mLST)-were compared with a novel broth designed for maximum recovery of E. sakazakii, E. sakazakii enrichment broth (ESE). One hundred seventy-seven strains (100%) grew in ESE, whereas between 2 and 6% of strains did not grow in EE, mLST, or ESSB. E. sakazakii possesses alpha-glucosidase activity, and a number of selective, chromogenic agars for E. sakazakii isolation based on this enzyme have been developed. E. sakazakii isolation agar produced fewer false-positive colonies than did Druggan-Forsythe-Iversen agar. However, the latter supported the growth of more E. sakazakii strains. It was also determined that 2% of E. sakazakii strains did not produce yellow pigmentation on tryptone soya agar at 25 degrees C, a characteristic frequently cited in the identification of E. sakazakii. The recovery of desiccated E. sakazakii (0.2 to 2000 CFU/25 g) from powdered IFM in the presence of a competing flora was determined with various enrichment broths and differential selective media. Current media designed for the isolation and presumptive identification of E. sakazakii do not support the growth of all currently known E. sakazakii phenotypes; therefore, improvements in the proposed methods are desirable.

  18. Effects of culture medium compositions on antidiabetic activity and anticancer activity of marine endophitic bacteria isolated from sponge

    Science.gov (United States)

    Maryani, Faiza; Mulyani, Hani; Artanti, Nina; Udin, Linar Zalinar; Dewi, Rizna Triana; Hanafi, Muhammad; Murniasih, Tutik

    2017-01-01

    High diversity of Indonesia marine spesies and their ability in producing secondary metabolite that can be used as a drug candidate cause this fascinating topic need to explore. Most of marine organisms explored to discover drug is macroorganism whereas microorganism (such as Indonesia marine bacteria) is very limited. Therefore, in this report, antidiabetic and anticancer activity of Indonesia marine bacteria isolated from Sponges's extract have been studied. Bacteria strain 8.9 which are collection of Research Center for Oseanography, Indonesian Institute of Sciences were from Barrang Lompo Island, Makasar, Indonesia. Bacteria were cultured in different culture medium compositions (such as: different pH, source of glucose and water) for 48 hours on a shaker, then they were extracted with ethyl asetate. Extracts of bacteria were tested by DPPH method (antioxidant activity), alpha glucosidase inhibitory activity method (antidiabetic activity), and Alamar Blue assay (anticancer activity) at 200 ppm. According to result, extract of bacteria in pH 8.0 exhibited the greatest antioxidant (19.27% inhibition), antidiabetic (63.95% inhibition) and anticancer activity of T47D cell line (44.62% cell viability) compared to other extracts. However, effect of addition of sugar sources (such as: glucose, sucrose, and soluble starch) and effect of addition of water/sea water exhibited less influence on their bioactivities. In conclusion, Indonesia marine bacteria isolated from sponge have potential a source of bioactive compound in drug discovery field.

  19. Recent Trends in Therapeutic Approaches for Diabetes Management: A Comprehensive Update

    Directory of Open Access Journals (Sweden)

    Pragya Tiwari

    2015-01-01

    Full Text Available Diabetes highlights a growing epidemic imposing serious social economic crisis to the countries around the globe. Despite scientific breakthroughs, better healthcare facilities, and improved literacy rate, the disease continues to burden several sections, especially middle and low income countries. The present trends indicate the rise in premature death, posing a major threat to global development. Scientific and technological advances have witnessed the development of newer generation of drugs like sulphonylureas, biguanides, alpha glucosidase inhibitors, and thiazolidinediones with significant efficacy in reducing hyperglycemia. Recent approaches in drug discovery have contributed to the development of new class of therapeutics like Incretin mimetics, Amylin analogues, GIP analogs, Peroxisome proliferator activated receptors, and dipeptidyl peptidase-4 inhibitor as targets for potential drugs in diabetes treatment. Subsequently, the identification and clinical investigation of bioactive substances from plants have revolutionized the research on drug discovery and lead identification for diabetes management. With a focus on the emerging trends, the review article explores the current statistical prevalence of the disease, discussing the benefits and limitations of the commercially available drugs. Additionally, the critical areas in clinical diabetology are discussed, with respect to prospects of statins, nanotechnology, and stem cell technology as next generation therapeutics and why the herbal formulations are consistently popular choice for diabetes medication and management.

  20. Drug Use Evaluation In Diabetic Patients at Out Patient Department Gorakhpur

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    Tripathi Poonam, Pandey Awanish, Pandey Rishabh, Goswamy Shambaditya & Srivatava Rashmi

    2010-12-01

    Full Text Available Objectives: The aim of the present study was to assess drug utilization in diabetes patients as a quantitative type of prescription auditing to generate data with respect to their extent variability of drug usage in a heath care system of the area of Gorakhpur.Material and Methods: The data for the study was collected from the survey conducted in February 2009-April 2009 at various hospitals of Gorakhpur. Two hundred diabetic patients were interviewed as a pre designed questionnaire based on clinical details. All the diabetic patients who visited the OPD during the study period enrolled in the study.Results: The pattern of drug prescription in diabetes showed that sulfonylurea (65% and biguanides (65% were most frequently prescribed followed by thiazolidinediones (23%and alpha glucosidase inhibitors (3%. Insulin was prescribed in 31 (15.5% patients. Among Most common co-existing illness was found hypertension (31.97 %. Majority of drugs were prescribed in oral dosage form (84.5 % followed by Parenteral (15.5 %.Conclusion: It is concluded that the present prescribing pattern of antidiabetic drugs Gorakhpur does not completely meet standard guidelines of diabetic treatment. Hence there is a need to encourage physicians of Gorakhpur to follow the guidelines while treating diabetes.

  1. Retrospective, Single Center Study of Clinical, Paraclinical and Natural Course of Infantile-Onset Pompe Disease

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    Noormohammad Noori

    2015-01-01

    Full Text Available Background: Infantile-onset Pompe disease is a rare genetic and lethal disorder which is caused by the lack of acid alpha-glucosidase activity (GAA. The aim of our study was to identify the demographic and clinical characteristics, and natural history of these patients. Materials and Methods: In this retrospective study, clinical file of 15 patients diagnosed with infantile-onset Pompe disease whose symptoms started before the age of 12 months were studied. Diagnosis was based on clinical history, physical examination and diagnostic parameters in chest X-ray, echocardiogram, electrocardiogram and biochemical tests after rule out the other metabolic and neuromuscular disorders. Results: Sixty percent of the patients were male and 40% were female. The mean age at the onset of symptoms was 78 days (range: 3-150 days. Most frequent clinical and paraclinical symptoms were cardiomegaly, hypotonia, hyporeflexia, macroglossia, failure to thrive, hepatomegaly, and feeding problems, respectively. The mean age at the time of death was 5.96 months (range: 4-8 months, and all patients died before one year of age. Muscle enzymes including AST, ALT, LDH, and CPK were elevated in all patients. Due to the lack of availability, enzyme replacement therapy was not possible for any patient. Conclusion: The study showed that despite the supportive measures and no specific treatment, the clinical course is not significantly different with similar studies and the overall prognosis of this form of disease is very poor and disappointing.

  2. IN VITRO α-AMYLASE AND α-GLUCOSIDASE INHIBITORY EFFECTS OF ETHANOLIC EXTRACT OF EVOLVULUS ALSINOIDES (L.

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    Duraisamy Gomathi

    2012-03-01

    Full Text Available Diabetes mellitus is a disorder in carbohydrate metabolism of endocrine system due to an absolute or relative deficiency of insulin secretion, action, or both. There has been an enormous interest in the development of alternative medicines for type 2 diabetes, specifically screening for phytochemicals with the ability to delay or prevent glucose absorption. Pancreatic α-amylase and glucosidase inhibitors offer an effective strategy to lower the levels of post prandial hyperglycemia via control of starch breakdown. The goal of the present study was to provide in vitro evidence for potential inhibition of α-glucosidase and α-amylase enzymes by using the ethanolic extract of Evolvulus alsinoides. The results showed a significant (more than 70% reduction in α –amylase activity as well as 50% reduction in α-glucosidase activity. The present study suggests that the extract of Evolvulus alsinoides effectively act as alpha amylase and alpha glucosidase inhibitor leading to a reduction in starch hydrolysis and hence eventually to loweredglucose levels.

  3. Effects of miglitol, sitagliptin or their combination on plasma glucose, insulin and incretin levels in non-diabetic men.

    Science.gov (United States)

    Aoki, Kazutaka; Masuda, Kiyomi; Miyazaki, Takashi; Togashi, Yu; Terauchi, Yasuo

    2010-01-01

    alpha-glucosidase inhibitors (alphaGIs) increase active glucagon-like peptide-1 (GLP-1) and reduce the total glucosedependent insulinotropic polypeptide (GIP) levels, but their ability to prevent diabetes remains uncertain. Dipeptidyl peptidase-4 (DPP-4) inhibitors, such as sitagliptin, increase active GLP-1 and GIP levels and improve hyperglycemia in a glucose-dependent fashion. However, the effectiveness of their combination in subjects with normal glucose tolerance (NGT) or impaired glucose tolerance (IGT) is uncertain. The present study evaluated the effect of miglitol, sitagliptin, and their combination on glucose, insulin and incretin levels in non-diabetic men. Miglitol and sitagliptin were administered according to four different intake schedules (C: no drug, M: miglitol; S: sitagliptin, M+S: miglitol and sitagliptin). The plasma glucose levels were significantly lower for M, S and M+S than for the control. The areas under the curve (AUCs) of the plasma active GLP-1 level in the M, S, and M+S groups were significantly greater than that in the control group. The AUC of the plasma active GLP-1 level was significantly greater for M+S group than for the M and S groups. The AUC of the plasma total GIP level was significantly smaller for M+S group than for the control and M and S groups. The results of our study suggest that miglitol, sitagliptin, or their combination contributes to the prevention of type 2 diabetes.

  4. Synthesis of alpha- and beta-D-glucopyranosyl triazoles by CuAAC 'click chemistry': reactant tolerance, reaction rate, product structure and glucosidase inhibitory properties.

    Science.gov (United States)

    Dedola, Simone; Hughes, David L; Nepogodiev, Sergey A; Rejzek, Martin; Field, Robert A

    2010-06-16

    Cu(I)-catalysed azide alkyne 1,3-dipolar cycloaddition (CuAAC) 'click chemistry' was used to assemble a library of 21 alpha-D- and beta-D-glucopyranosyl triazoles, which were assessed as potential glycosidase inhibitors. In the course of this work, different reactivities of isomeric alpha- and beta-glucopyranosyl azides under CuAAC conditions were noted. This difference was further investigated using competition reactions and rationalised on the basis of X-ray crystallographic data, which revealed significant differences in bond lengths within the azido groups of the alpha- and beta-anomers. Structural studies also revealed a preference for perpendicular orientation of the sugar and triazole rings in both the alpha- and beta-glucosyl triazoles in the solid state. The triazole library was assayed for inhibition of sweet almond beta-glucosidase (GH1) and yeast alpha-glucosidase (GH13), which led to the identification of a set of glucosidase inhibitors effective in the 100 microM range. The preference for inhibition of one enzyme over the other proved to be dependent on the anomeric configuration of the inhibitor, as expected.

  5. Australine, a pyrrolizidine alkaloid that inhibits amyloglucosidase and glycoprotein processing

    Energy Technology Data Exchange (ETDEWEB)

    Tropea, J.E.; Molyneux, R.J.; Kaushal, G.P.; Pan, Y.T.; Mitchell, M.; Elbein, A.D. (Univ. of Texas Health Science Center, San Antonio (USA))

    1989-03-07

    Australine is a polyhydroxylated pyrrolizidine alkaloid that was isolated from the seeds of the Australian tree Castanospermum australe and characterized by NMR and X-ray diffraction analysis. Since swainsonine and catanospermine are polyhydroxylated indolizidine alkaloids that inhibit specific glycosidases, the authors tested australine against a variety of exoglycosidases to determine whether it would inhibit any of these enzymes. This alkaloid proved to be a good inhibitor of the {alpha}-glucosidase amyloglucosidase (50% inhibition at 5.8 {mu}M), but it did not inhibit {beta}-glucosidase, {alpha}- or {beta}-mannosidase, or {alpha}- or {beta}-galactosidase. The inhibition of amyloglucosidase was of a competitive nature. Australine also inhibited the glycoprotein processing enzyme glucosidase I, but had only slight activity toward glucosidase II. When incubated with cultured cells, this alkaloid inhibited glycoprotein processing at the glucosidase I step and caused the accumulation of glycoproteins with Glc{sub 3}Man{sub 7-9}(GlcNAc){sub 2}-oligosaccharides.

  6. Pathological features of glycogen storage disease type II highlighted in the knockout mouse model.

    Science.gov (United States)

    Bijvoet, A G; Van Hirtum, H; Vermey, M; Van Leenen, D; Van Der Ploeg, A T; Mooi, W J; Reuser, A J

    1999-11-01

    Glycogen storage disease type II (GSDII; Pompe's disease) is an autosomal recessive disease caused by lysosomal alpha-glucosidase deficiency. Skeletal muscle weakness is the most conspicuous clinical symptom of patients suffering from GSDII and skeletal muscle also is prominently involved in the knockout mouse model of this disease. Thus far, however, little detailed information has been published on the pathological changes in other mouse tissues. This paper aims to provide these data and gives a record of the clinical course of the mouse model over a 2-year period. Four-month-old affected mice perform worse in a running wheel than their unaffected littermates, but do not yet display other clear signs of disease. The lysosomal glycogen storage, already evident at birth, becomes more severe in time, leading to muscle wasting by 9-10 months of age and then limb girdle weakness and kyphosis. The disease does not markedly shorten the animal's life span despite the serious tissue pathology, which is not limited to heart and skeletal muscle, but is also seen in the smooth muscle of blood vessels and of the respiratory, digestive, and urogenital tracts. In addition, the mice have lysosomal glycogen storage in the liver, kidney, spleen, and salivary gland; in Schwann cells of the peripheral nerves, and in a subset of neurons in the central nervous system. By pathological criteria, the knockout mouse model parallels the human infantile form of GSDII and is attractive for studying the possible reversal of tissue pathology and symptomatology under different therapeutic regimes.

  7. Late-onset Pompe disease with complicated intracranial aneurysm: a Chinese case report

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    Zhang B

    2016-03-01

    Full Text Available Bin Zhang,1,2,* Yuying Zhao,1,3,* Junling Liu,1,4 Ling Li,1 Jingli Shan,1 Dandan Zhao,1 Chuanzhu Yan1,3 1Laboratory of Neuromuscular Disorders and Department of Neurology, Qilu Hospital of Shandong University, Jinan, Shandong, 2Department of Neurology, Liaocheng People’s Hospital, Liaocheng, Shandong, 3Department of Neurology, Qilu Hospital of Shandong University, Key Laboratory for Experimental Teratology of the Ministry of Education, Brain Science Research Institute, Shandong University, Jinan, Shandong, 4Department of Neurology, Affiliated Hospital of Weifang Medical University, Weifang, Shandong, People’s Republic of China *These authors contributed equally to this work Abstract: Pompe disease is a rare autosomal recessive hereditary disease caused by genetic defects of acid maltase. This disease could be divided into two forms: infantile and late-onset, which mainly affect cardiac, respiratory, and skeletal muscle systems. Late-onset patients mainly show symptoms of skeletal muscle involvement, but recent reports have found that the central nervous system was also affected in some patients. Herein, we report a case of a female, adolescent-onset Pompe patient, who was diagnosed with complicated intracranial aneurysm in adulthood. Keywords: Pompe disease, glycogen storage disease II, acid maltase, acid alpha-glucosidase, cerebrovascular disorders

  8. A quick overview on some aspects of endocrinological and therapeutic effects of Berberis vulgaris L.

    Directory of Open Access Journals (Sweden)

    Ali Zarei

    2015-10-01

    Full Text Available Many herbaceous plants contain compounds that have biological effects in addition to their medicinal properties. They have compounds with numerous properties, including hypo lipidemic, hypoglycemic, antioxidant, and hepato protective ones, which have been analyzed at different levels. One of these plants, with the scientific name of Berberis vulgaris, is barberry. The most important compounds identified in this plant are berberine, oxycontin, palmatine, bervulcine, berbamine, columbamine, jatrorrhizine, coptisine, and berbamine. In addition to alkaloids, organic acids such as chelidonic acid, citric acid, malic acid, resin, tannin, pectinic, and mucilagic substances are among the ingredients of barberry. In this paper, it was attempted to determine the role and effect of the extract of barberry on various body organs. The results showed that berberine actually increases insulin sensitivity and is capable of inhibiting alpha glucosidase, adipogenesis, and thus acts as an anti-obesity and hypoglycemic agent. Berberine reduces the density of serum cholesterol and triglycerides and can improve the function of liver enzymes, therefore, it can be suggested as a hypo lipidemic and hepato protective plant extract. The hepato protective effects of this extract are probably due to its antioxidant properties. Studies showed that barberry have numerous health benefits, including anti-inflammatory ones. Moreover, it can be used as a medicinal herb to treat a variety of disorders, such as diabetes, liver disease, gallbladder pain, digestive, urinary tract diseases, and gallstones. However, more studies on this issue and doing more focused and intensive researches in this field are recommended.

  9. Targeted screening for the detection of Pompe disease in patients with unclassified limb-girdle muscular dystrophy or asymptomatic hyperCKemia using dried blood: A Spanish cohort.

    Science.gov (United States)

    Gutiérrez-Rivas, E; Bautista, J; Vílchez, J J; Muelas, N; Díaz-Manera, J; Illa, I; Martínez-Arroyo, A; Olivé, M; Sanz, I; Arpa, J; Fernández-Torrón, R; López de Munáin, A; Jiménez, L; Solera, J; Lukacs, Z

    2015-07-01

    We aimed to screen for Pompe disease in patients with unclassified limb-girdle muscular dystrophy (LGMD) or asymptomatic hyperCKemia using dried blood spot (DBS) assays. Subsequently, we aimed to calculate the diagnostic delay between initial symptom presentation and the diagnosis. A prospective, multicenter, observational study was conducted in 348 patients: 146 with unclassified LGMD and 202 with asymptomatic or paucisymptomatic hyperCKemia. We quantified levels of acid alpha-glucosidase (GAA) from dried blood spots analyzed fluorometrically. The test was positive in 20 patients, and Pompe disease was confirmed by genetic testing in 16. Undiagnosed Pompe disease was detected in 7.5% of patients with LGMD and in 2.5% of patients with persistent, idiopathic elevation of serum creatine kinase. The c.-32-13 T > G mutation was found most commonly. The diagnostic delay was 15 years on average. In conclusion, DBS tests are useful and reliable screening tools for Pompe disease. We recommend the dried blood spot test to be included in the diagnostic work-up of patients with unclassified myopathies with proximal weakness and/or hyperCKemia of unknown cause and, when positive, to define the diagnosis, it will have to be confirmed by biochemical and/or molecular genetic analysis.

  10. Involvement of Kytococcus schroeteri in a case of prosthetic valve endocarditis

    Directory of Open Access Journals (Sweden)

    Antonella D’Andria

    2008-03-01

    Full Text Available We report a fourth case of prosthetic valve endocarditis caused by the newly described micrococcal species: Kytococcus schroeteri. A 38-year old male was admitted to our hospital with suspected prosthetic valve endocarditis. In three blood cultures and in the prosthetic heart valve culture grew the same type of microrganism, initially identified as Micrococcus luteus. Presuntive assignment to the genus Kytococcus was suggested by the arginina dihydrolase activity and by the resistence to penicillin, oxacillin and methicillin, characteristics which are not shared by other micrococci. Further biochemical tests confirmed that the isolates belonged to the genus Kytococcus (negative reaction for oxydase, esculin and urease, positive for catalase and arginine dihydrolase species schroeteri (alpha-glucosidase positive. The patient was treated with vancomycin, gentamicin and rifampicin. After the valve replacement, on the 20th day of hospitalization, the patient’s biological parameters returned to normal values, and after one mounth the resolution of the disease was completed. The diagnosis of four cases by K. schroeteri endocarditis, described within a short period of time (2003-2006, might indicate a specific pathogenicity of this new species. In case of endocarditis diagnosis, the suspect of the genus Kytococcus involvement should be considered each time the aerobic Gram positive cocci, with arginina dihidrolase activiy and the resistence to penicillin, oxacillin and methicillin, are isolated by significative biological samples.

  11. Attachment of MAL32-encoded maltase on the outside of yeast cells improves maltotriose utilization.

    Science.gov (United States)

    Dietvorst, J; Blieck, L; Brandt, R; Van Dijck, P; Steensma, H Y

    2007-01-01

    The fermentation of maltotriose, the second most abundant fermentable sugar in wort, is often incomplete during high-gravity brewing. Poor maltotriose consumption is due to environmental stress conditions during high-gravity fermentation and especially to a low uptake of this sugar by some industrial strains. In this study we investigated whether the use of strains with an alpha-glucosidase attached to the outside of the cell might be a possible way to reduce residual maltotriose. To this end, the N-terminal leader sequence of Kre1 and the carboxy-terminal anchoring domain of either Cwp2 or Flo1 were used to target maltase encoded by MAL32 to the cell surface. We showed that Mal32 displayed on the cell surface of Saccharomyces cerevisiae laboratory strains was capable of hydrolysis of alpha-1,4-linkages, and that it increased the ability of a strain lacking a functional maltose permease to grow on maltotriose. Moreover, the enzyme was also expressed and found to be active in an industrial strain. These data show that expressing a suitable maltase on the cell surface might provide a means of modifying yeast for more complete maltotriose utilization in brewing and other fermentation applications.

  12. Fractures in children with Pompe disease: a potentiallong-term complication

    Energy Technology Data Exchange (ETDEWEB)

    Case, Laura E. [Duke University Medical Center, Division of Physical Therapy, Department of Community and Family Medicine, Durham, NC (United States); Hanna, Rabi; DeArmey, Stephanie; Mackey, Joanne; Boney, Anne; Chen, Yuan-Tsong; Kishnani, Priya S. [Duke University Medical Center, Department of Pediatrics, Durham, NC (United States); Frush, Donald P. [Duke University Medical Center, Department of Radiology, Durham, NC (United States); Krishnamurthy, Vidya [Duke University Medical Center, Department of Pediatrics and Department of Medicine, Durham, NC (United States); Morgan, Claire; Bouchard, Susan [Genzyme Corporation, Pharmacovigilance, Cambridge, MA (United States); Corzo, Deyanira [Genzyme Corporation, Cambridge, MA (United States); Weber, Thomas J. [Duke University Medical Center, Department of Medicine, Durham, NC (United States)

    2007-05-15

    Pompe disease (glycogen storage disease type II or acid maltase deficiency) is an autosomal recessive disorder caused by deficiency of the lysosomal enzyme acid {alpha}-glucosidase (GAA). Classic infantile-onset disease, characterized by cardiomegaly and profound weakness, leads to death in the first year of life from cardiorespiratory failure. Reversal of cardiomyopathy and improved motor function have been shown in clinical trials of rhGAA enzyme replacement therapy (ERT) with alglucosidase alfa (Myozyme), recently approved for clinical use. Increased survival potentially unmasks long-term complications of this previously lethal disease, including risk of skeletal fracture, recently identified at our institution and not previously reported in children with Pompe disease. To report the risk of fracture in children with Pompe disease with increased survival with ERT. We present four cases of fracture in patients with classic infantile Pompe disease treated with ERT at our institution, and review a study database for additional reports of fracture in this population. We review 19 fractures in 14 children with Pompe disease on ERT. Radiologists should be familiar with and vigilant for the association of fractures and increased survival on ERT in children with Pompe disease. We discuss potential mechanisms, implications for radiographic surveillance, potential intervention, and needs for further research. (orig.)

  13. A unique de novo interstitial deletion of chromosome 17, del(17)(q23.2q24.3) in a female newborn with multiple congenital anomalies

    Energy Technology Data Exchange (ETDEWEB)

    Levin, M.L.; Shaffer, L.G.; Lewis, R.A. [Baylor College of Medicine, Houston, TX (United States)] [and others

    1994-09-01

    Contiguous gene or microdeletion syndromes occurring on chromosome 17p include the Smith-Magenis and Miller-Dieker syndromes associated with interstitial deletions of 17p11.2 and 17p13.3, respectively. Other cytogenetically visible interstitial deletions on chromosome 17 are quite rare or unique. We describe a newborn with a novel interstitial deletion of the long arm of chromosome 17 [del(17)(q23.2q24.3)] who died on day of life 17 during a recurrent apneic episode. We have compared our patient`s phenotype and karyotype to two reported patients with deletion 17q with minor clinical overlap. The most striking clinical features of this patient were severe intrauterine growth retardation, widespread skeletal malformations (split sutures, hypoplastic acetabulae and scapulae, vertebral anomalies, and digital hypoplasia), cutis verticis gyrata, dysmorphic facial features, and oropharyngeal malformations (absent uvula and submucous cleft palate). Mild congenital heart disease and anomalous optic nerves were also present. Parental karyotyps were normal. DNA from parents and patient has been collected and cell lines established on both parents. Genes which have been previously mapped to the region that is apparently deleted in this patient include: chorionic somatomammotropin A, growth hormone (normal), acid alpha-glucosidase, apolipoprotein H, and the alpha peptide of type 4 voltage gated sodium channel. As in other clinical cytogenetic syndromes, further descriptions of patients with similar or overlapping rearrangements in this region will be necessary to delineate genotype/phenotype correlations for chromosome 17.

  14. Clinical Analysis of Algerian Patients with Pompe Disease

    Science.gov (United States)

    Medjroubi, M.; Froissart, R.; Taghane, N.; Sifi, K.; Benhabiles, A.; Lemai, S.; Semra, S.; Benmekhebi, H.; Bouderda, Z.; Abadi, N.; Hamri, A.

    2017-01-01

    Pompe's disease is a metabolic myopathy caused by a deficiency of acid alpha-glucosidase (GAA), also called acid maltase, an enzyme that degrades lysosomal glycogen. The clinical presentation of Pompe's disease is variable with respect to the age of onset and rate of disease progression. Patients with onset of symptoms in early infancy (infantile-onset Pompe disease (IOPD)) typically exhibit rapidly progressive hypertrophic cardiomyopathy and marked muscle weakness. Most of them die within the first year of life from cardiac and/or respiratory failure. In the majority of cases of Pompe's disease, onset of symptoms occurs after infancy, ranging widely from the first to sixth decade of life (late-onset Pompe's disease or LOPD). Progression of the disease is relentless and patients eventually progress to loss of ambulation and death due to respiratory failure. The objective of this study was to characterize the clinical presentation of 6 patients (3 with EOPD and the other 3 with LOPD) of 5 families from the East of Algeria. All our patients were diagnosed as having Pompe's disease based on biochemical confirmations of GAA deficiency by dried blood spots (DBS) and GAA gene mutations were analyzed in all patients who consented (n = 4). Our results are similar to other ethnic groups. PMID:28265479

  15. Effect of tacrolimus on the cauda epididymis in rats: analysis of epididymal biochemical markers or antioxidant defense enzymes.

    Science.gov (United States)

    Hisatomi, Akihiko; Sakuma, Shozo; Fujiwara, Michio; Seki, Jiro

    2008-01-14

    The effect of tacrolimus on epididymal biochemical markers was investigated following single daily subcutaneous doses of 1, 2 and 3 mg kg(-1) day(-1) for 2 weeks to male adult rats. The tacrolimus 2 and 3 mg kg(-1) day(-1) groups showed a significant and dose-dependent decrease in sperm count in the cauda epididymis. Among tissue levels of L-carnitine, alpha-glucosidase and acid phosphatase, only L-carnitine level in the cauda epididymis was significantly reduced in the tacrolimus 3 mg kg(-1)day(-1) group. However, no significant difference was seen in the plasma L-carnitine. It was suggested that lowering of L-carnitine in the cauda epididymis was attributable to the adverse effect on epididymal function to transport and/or concentrate L-carnitine. Since L-carnitine has been reported to have antioxidant potential, antioxidant defense enzymes in the cauda epididymis such as superoxide dismutase (SOD), catalase, glutathione peroxidase and glutathione reductase were evaluated. The results showed no significant differences in activities, confirming that the treatment with tacrolimus did not affect the activities of these antioxidant enzymes. In conclusion, this study indicates that tacrolimus induces a decrease in L-carnitine level in the cauda epididymis, which is probably caused by impairment of epididymal function to transport and/or concentrate L-carnitine from bloodstream, and a decrease in sperm count.

  16. Voglibose administration before the evening meal improves nocturnal hypoglycemia in insulin-dependent diabetic patients with intensive insulin therapy.

    Science.gov (United States)

    Taira, M; Takasu, N; Komiya, I; Taira, T; Tanaka, H

    2000-04-01

    Nocturnal hypoglycemia is one of the serious complications of intensive insulin therapy in patients with insulin-dependent diabetes mellitus (IDDM; type 1 DM). We assessed the effect of voglibose (alpha-glucosidase inhibitor) administration before the evening meal on nocturnal hypoglycemia in IDDM patients with intensive insulin therapy. Ten IDDM patients received 0.3 mg voglibose just before the evening meal for 5 days. The diet and insulin regimen were not changed throughout the study. Nocturnal plasma glucose levels (10 PM, 3 AM, and 7 AM) were studied in these patients before and during voglibose administration. Blood glucose levels were measured at 3 AM before and during voglibose treatment. The mean plasma glucose level at 3 AM was 3.4+/-0.4 mmol/L before voglibose treatment and 7.3+/-1.0 mmol/L during treatment. Plasma glucose at 3 AM was elevated in 9 of 10 patients with voglibose. The decrease in plasma glucose from 10 PM to 3 AM was 6.5+/-0.8 mmol/L before voglibose administration but 3.2+/-0.9 mmol/L during treatment (P hypoglycemia rate was 52% (17 of 33 nights) before voglibose administration but only 9.1% (3 of 33 nights) during treatment. We conclude that voglibose administration before the evening meal improves nocturnal hypoglycemia in IDDM patients with intensive insulin therapy.

  17. α-Glucosidase Inhibitory Activity of Polyphenols from the Burs of Castanea mollissima Blume

    Directory of Open Access Journals (Sweden)

    Jianwei Zhang

    2014-06-01

    Full Text Available Polyphenol extracts from the burs of Castanea mollissima Blume (CMPE exhibited potential antioxidant and hypoglycemic activities. The α-glucosidase inhibitory activities of CMPE were assessed as a means of elucidating the mechanism behind its hypoglycemic activities. In vitro studies showed that CMPE significantly inhibited both yeast α-glucosidase, through a noncompetitive mode with an IC50 of 0.33 μg/mL, and rat intestinal α-glucosidase. In vivo studies revealed that oral administration of CMPE at doses of 600 mg/kg significantly reduced postprandial blood glucose levels by 27.2% in normal rats following sucrose challenges. Gel permeation chromatography revealed that CMPE exhibited typical characteristics of high-molecular-mass polymers with mean (Mn and weight (Mw average molecular weights of 35.4 and 50.7 kDa, respectively, and a polydispersity (Mw/Mn of 1.432. Acid hydrolysis analysis indicated the presence of ellagitannins. These data suggest that CMPE, enriched with ellagitannins, would be an efficacious dietary supplement for diabetes management through the inhibition of alpha-glucosidase.

  18. Inhibition of Porcine Pancreatic Amylase Activity by Sulfamethoxazole: Structural and Functional Aspect.

    Science.gov (United States)

    Maity, Sujan; Mukherjee, Koel; Banerjee, Amrita; Mukherjee, Suman; Dasgupta, Dipak; Gupta, Suvroma

    2016-06-01

    Combating Type-2 diabetes mellitus is a pivotal challenge in front of the present world. Several lines of therapy are in practice for resisting this deadly disease which often culminates with cardiovascular complexities, neuropathy and retinopathy. Among various therapies, administration of alpha glucosidase inhibitors is common and widely practiced. Sulfonylurea category of anti diabetic drug often suffers from cross reactivity with sulfamethoxazole (SMX), a common drug in use to treat a handful of microbial infections. However the specific cellular target generating postprandial hypoglycemia on SMX administration is till date unraveled. The present work has been initiated to elucidate the effects of a group of sulfonamide drugs inclusive of SMX for their amylase inhibitory role. SMX inhibits porcine pancreatic amylase (PPA) in a noncompetitive mode with an average IC50 value 0.94 mM respectively. Interaction of SMX with PPA is manifested with gradual quenching of tryptophan fluorescence with concomitant shift in lambda max value (λmax). Binding is governed by entropy driven factor (24.8 cal mol(-1) K(-1)) with unfavorable contribution from enthalpy change. SMX interferes with the activity of acarbose in a synergistic mode to reduce the effective dose of acarbose as evident from the in vitro PPA inhibition study. In summary, loss of PPA activity in presence of SMX is indicative of structural changes of PPA which is further augmented in the presence of acarbose as explained in the schematic model and docking study.

  19. New α-Glucosidase Inhibitory Triterpenic Acid from Marine Macro Green Alga Codium dwarkense Boergs

    Directory of Open Access Journals (Sweden)

    Liaqat Ali

    2015-07-01

    Full Text Available The marine ecosystem has been a key resource for secondary metabolites with promising biological roles. In the current study, bioassay-guided phytochemical investigations were carried out to assess the presence of enzyme inhibitory chemical constituents from the methanolic extract of marine green alga—Codium dwarkense. The bioactive fractions were further subjected to chromatographic separations, which resulted in the isolation of a new triterpenic acid; dwarkenoic acid (1 and the known sterols; androst-5-en-3β-ol (2, stigmasta-5,25-dien-3β,7α-diol (3, ergosta-5,25-dien-3β-ol (4, 7-hydroxystigmasta-4,25-dien-3-one-7-O-β-d-fucopyranoside (5, 7-hydroxystigmasta-4,25-dien-3-one (6, and stigmasta-5,25-dien-3β-ol (7. The structure elucidation of the new compound was carried out by combined mass spectrometry and 1D (1H and 13C and 2D (HSQC, HMBC, COSY, and NOESY NMR spectroscopic data. The sub-fractions and pure constituents were assayed for enzymatic inhibition of alpha-glucosidase. Compound 1 showed significant inhibition at all concentrations. Compounds 2, 3, 5, and 7 exhibited a dose-dependent response, whereas compounds 4–6 showed moderate inhibition. Utilizing such marine-derived biological resources could lead to drug discoveries related to anti-diabetics.

  20. Effects of MAL61 and MAL62 overexpression on maltose fermentation of baker's yeast in lean dough.

    Science.gov (United States)

    Zhang, Cui-Ying; Lin, Xue; Song, Hai-Yan; Xiao, Dong-Guang

    2015-08-01

    The predominant fermentable sugar in lean dough is maltose. To improve the leavening ability of baker's yeast in lean dough, maltose metabolism should be improved. Maltase (alpha-glucosidase, encoded by MAL62) and maltose permease (encoded by MAL61) are the major factors involved in maltose metabolism. The major rate-limiting factor in maltose metabolism and leavening ability of baker's yeast remains unclear. In this work, MAL61 and/or MAL62 overexpression strains were constructed to investigate the decisive factor for maltose metabolism of industrial baker's yeast in lean dough. Our results show that elevated maltose permease activity by MAL61 overexpression yielded less improvement in maltose fermentation compared to elevated maltase activity by MAL62 overexpression. Significant increase in maltase activity by MAL62 overexpression could result in a 44% increase in leavening ability of industrial baker's yeast in lean dough and a 39% increase in maltose metabolism in a medium containing glucose and maltose. Thus, maltase was the rate-limiting factor in maltose fermentation of industrial baker's yeast in lean dough. This study lays a foundation for breeding of industrial baker's yeast for quick dough leavening.

  1. A cardiologic approach to non-insulin antidiabetic pharmacotherapy in patients with heart disease

    Directory of Open Access Journals (Sweden)

    Fisman Enrique Z

    2009-07-01

    Full Text Available Abstract Classical non-insulin antihyperglycemic drugs currently approved for the treatment of type 2 diabetes mellitus (T2DM comprise five groups: biguanides, sulfonylureas, meglitinides, glitazones and alpha-glucosidase inhibitors. Novel compounds are represented by the incretin mimetic drugs like glucagon like peptide-1 (GLP-1, the dipeptidyl peptidase 4 (DPP-4 inhibitors, dual peroxisome proliferator-activated receptors (PPAR agonists (glitazars and amylin mimetic drugs. We review the cardiovascular effects of these drugs in an attempt to improve knowledge regarding their potential risks when treating T2DM in cardiac patients. Metformin may lead to lethal lactic acidosis, especially in patients with clinical conditions that predispose to this complication, such as recent myocardial infarction, heart or renal failure. Sulfonylureas exert their effect by closing the ATP-dependent potassium channels. This prevents the opening of these channels during myocardial ischemia, impeding the necessary hyperpolarization that protects the cell. The combined sulfonylurea/metformin therapy reveals additive effects on mortality in patients with coronary artery disease (CAD. Meglitinides effects are similar to those of sulfonylureas, due to their almost analogous mechanism of action. Glitazones lower leptin levels, leading to weight gain and are unsafe in NYHA class III or IV. The long-term effects of alpha-glucosidase inhibitors on morbidity and mortality rates is yet unknown. The incretin GLP-1 is associated with reductions in body weight and appears to present positive inotropic effects. DPP-4 inhibitors influences on the cardiovascular system seem to be neutral and patients do not gain weight. The future of glitazars is presently uncertain following concerns about their safety. The amylin mimetic drug paramlintide, while a satisfactory adjuvant medication in insulin-dependent diabetes, is unlikely to play a major role in the management of T2DM

  2. Hypoglycemic agents and potential anti-inflammatory activity

    Directory of Open Access Journals (Sweden)

    Kothari V

    2016-04-01

    Full Text Available Vishal Kothari,1 John A Galdo,2 Suresh T Mathews3 1Department of Nutrition and Dietetics, Boshell Diabetes and Metabolic Diseases Research Program, Auburn University, Auburn, 2Department of Pharmacy Practice, 3Department of Nutrition and Dietetics, Samford University, Birmingham, AL, USA Abstract: Current literature shows an association of diabetes and secondary complications with chronic inflammation. Evidence of these immunological changes include altered levels of cytokines and chemokines, changes in the numbers and activation states of various leukocyte populations, apoptosis, and fibrosis during diabetes. Therefore, treatment of diabetes and its complications may include pharmacological strategies to reduce inflammation. Apart from anti-inflammatory drugs, various hypoglycemic agents have also been found to reduce inflammation that could contribute to improved outcomes. Extensive studies have been carried out with thiazolidinediones (peroxisome proliferator-activated receptor- agonist, dipeptidyl peptidase-4 inhibitors, and metformin (AMP-activated protein kinase activator with each of these classes of compounds showing moderate-to-strong anti-inflammatory action. Sulfonylureas and alpha glucosidase inhibitors appeared to exert modest effects, while the injectable agents, insulin and glucagon-like peptide-1 receptor agonists, may improve secondary complications due to their anti-inflammatory potential. Currently, there is a lack of clinical data on anti-inflammatory effects of sodium–glucose cotransporter type 2 inhibitors. Nevertheless, for all these glucose-lowering agents, it is essential to distinguish between anti-inflammatory effects resulting from better glucose control and effects related to intrinsic anti-inflammatory actions of the pharmacological class of compounds. Keywords: diabetes, inflammation, insulin, metformin, thiazolidinedione, gliptin

  3. Acute effects of acarbose on post-prandial glucose and triglycerides in type 2 diabetics following intake of different Malaysian foods.

    Science.gov (United States)

    Nawawi, H M; Yazid, T N; Ismail, F; Khalid, B A

    2000-03-01

    Acarbose inhibits intestinal alpha-glucosidases resulting in diminished and delayed postprandial hyperglycaemia (PPH). Studies on effects of acarbose on postprandial lipaemia (PPL) have been inconclusive. Little is known about the effects of acarbose on PPH and PPL following intake of a polysaccharide diet. We studied 30 type 2 diabetic patients on dietary and/or oral hypoglycaemic agent(s). Thirty patients were recruited for food A (nasi lemak), 28 for food B (mee goreng) and 28 for food C (roti telur), which represent the typical diets of the three main races in Malaysia. Serial blood samples were taken at 15 min before and up to 240 min after each food intake, without acarbose. Subsequently, three doses of 50 mg acarbose were given orally and the same procedure was repeated the following day. There were significantly lower mean increments in plasma glucose levels after compared to before acarbose treatment 30, 45 and 60 min for food A and at 30, 45, 60, 120, 180 and 240 min for food C, but no significant difference was noted for food B. There was a significantly lower mean fasting glucose level after compared with before acarbose treatment following intake of food A and C but not food B. Short-term treatment with acarbose caused significant diminished and delayed PPH response with food A and C but not with food B. Acarbose was more effective in reducing PPH response in polysaccharide foods with a higher and earlier postprandial glucose peak than in those with a lower and lagged peak. There were no significant differences in the mean fasting or postprandial triglyceride levels before and after acarbose treatment, following intake of all three foods for up to 4 hours. Depending on the food absorption pattern, overnight low dose treatment with acarbose leads to diminished fasting and peak plasma glucose levels, and delayed PPH but insignificant reduction in postprandial lipaemia in poorly controlled type 2 diabetics following intake of racially different Malaysian

  4. Activities of starch hydrolytic enzymes and sucrose-phosphate synthase in the stems of rice subjected to water stress during grain filling.

    Science.gov (United States)

    Yang, J; Zhang, J; Wang, Z; Zhu, Q

    2001-11-01

    To understand the effect of water stress on the remobilization of prestored carbon reserves, the changes in the activities of starch hydrolytic enzymes and sucrose-phosphate synthase (SPS) in the stems of rice (Oryza sativa L.) during grain filling were investigated. Two rice cultivars, showing high lodging-resistance and slow remobilization, were grown in the field and subjected to well-watered (WW, psi(soil)=0) and water-stressed (WS, psi(soil)=-0.05 MPa) treatments 9 d after anthesis (DAA) till maturity. Leaf water potentials of both cultivars markedly decreased during the day as a result of WS treatment, but completely recovered by early morning. WS treatment accelerated the reduction of starch in the stems, promoted the reallocation of prefixed (14)C from the stems to grains, shortened the grain filling period, and increased the grain filling rate. More soluble sugars including sucrose were accumulated in the stems under WS than under WW treatments. Both alpha- and beta-amylase activities were enhanced by the WS, with the former enhanced more than the latter, and were significantly correlated with the concentrations of soluble sugars in the stems. The other two possible starch-breaking enzymes, alpha-glucosidase and starch phosphorylase, showed no significant differences in the activities between the WW and WS treatments. Water stress also increased the SPS activity that is responsible for sucrose production. Both V(limit) and V(max), the activities of the enzyme at limiting and saturating substrate concentrations, were enhanced and the activation state (V(limit)/V(max)) was also increased as a result of the more significant enhancement of V(limit). The enhanced SPS activity was closely correlated with an increase of sucrose accumulation in the stems. The results suggest that the fast hydrolysis of starch and increased carbon remobilization were attributed to the enhanced alpha-amylase activity and the high activation state of SPS when the rice was subjected

  5. ANTI-OXIDANT AND ENZYME-INHIBITORY POTENTIAL OF MARINE STREPTOMYCES

    Directory of Open Access Journals (Sweden)

    K. Suthindhiran

    2013-01-01

    Full Text Available Marine actinomycetes are potential source for the discovery of novel compounds and enzymes. Though extensive research on marine actinomycetes is underway globally, the actinomycetes research from Indian marine ecosystem is unexplored and understudied. Hence, the present research is focussed on the screening of bioactive compounds from marine actinomycetes isolated from Indian coastal region. This study is designed to determine the antioxidant and enzyme inhibitory potential of Streptomyces sp. VITMSS05 strain, isolated from Marakkanam, southern coast of India. An actinomycetes strain designated as VITMSS05 was isolated. This strain was cultivated in Starch Caesin Agar medium (SCA supplemented with sea water. The cultural, morphological and molecular characterization was determined for the isolate. The crude extract of the isolate was extracted with ethyl acetate. Antioxidant activity of the crude extract was determined by DPPH radical scavenging assay. Alpha amylase and alpha glucosidase inhibitory potential of the extract was determined. Based on the phenotypic and phylogenetic analysis the strain was identified as Streptomyces sp. Significant antioxidant activity of the extract was observed with an IC50 value of 92.49 μg mL-1. The extract shows 64.1% inhibition on α-amylase and 91.5% inhibition on α-glucosidase at 100 μg mL-1 with an IC50 value of 385.97 and 42.89 μg mL-1. From the results it is evident that the ethyl acetate extract of Streptomyces sp. VITMSS05 has potent antioxidant and enzyme inhibitory activity in vitro. The combined effect of free radical scavenging and enzyme inhibition makes it a potent anti diabetic drug.

  6. The inhibitory effects on adult male reproductive functions of crude garlic (Allium sativum) feeding

    Institute of Scientific and Technical Information of China (English)

    Imen Hammami; Afef Nahdi; Claire Mauduit; Mohamed Benahmed; Mohamed Amri; Awatef Ben Amar; Semy Zekri; Ahmed El May; Michele Veronique El May

    2008-01-01

    Aim: To investigate the effects of crude garlic on adult male rat reproductive functions. Methods: Thirty male rats were divided into five groups: group 1 (untreated) and groups 2, 3, 4 and 5 were fed for 30 days with 5%, 10%, 15% and 30% crude garlic, respectively. Testes and accessory organs were weighed and some markers were assessed. Light and electron microscopy observations were also performed. Results: A significant decrease was observed in the body weight of groups 4 (14%; P < 0.01) and 5 (20%; P < 0.01); of the prostate weight in group 5 (29.1%; P < 0.05) and of seminal vesicle weight in groups 3 (14.4%; P < 0.01), 4 (18.3%; P < 0.01) and 5 (27.3%; P < 0.01). In contrast, testis and epididymis weights were unchanged. In epididymis tissue, the alpha glucosidase activity and the spermatozoa density were unchanged. The treatment resulted in a significant decrease in testosterone serum levels in groups 3 (77.3%; P < 0.01), 4 (77.3%; P < 0.01) and 5 (90.9%; P < 0.01), associated with a significant increase in LH serum levels (P < 0.01). Testicular histology showed a dose-dependent increase in the percentage of empty seminiferous tubules. Moreover, testicular function was affected; a significant decrease in phosphatase acid activity (P < 0.01) and testosterone (P < 0.05) contents were observed. Conclusion: Crude garlic consumption during 1 month reduced testosterone secretion and altered spermatogenesis at 10%, 15% and 30% doses. (Asian J Androl 2008 Jul; 10: 593-601)

  7. Composition and Biological Activity of Volatile Oil from Salviajudaica and S. multicaulis from Jordan.

    Science.gov (United States)

    Afifi, Fatma U; Kasabri, Violet; Al-Jaber, Hala I; Abu-Irmaileh, Barakat E; Al-Qudah, Mahmoud A; Abazaa, Ismail F

    2016-04-01

    The aim of this work was to determine the composition of the hydro-distilled essential oil of Salvia judaica Boiss. and S. multicaulis Vahl. (Lamiaceae) from Jordan by GC and GC-MS and to report the actual composition of their fresh leaves and flowers using SPME (Solid Phase Micro-Extraction).Their dual alpha-amylase/alpha glucosidase and pancreatic lipase inhibitory activities as well as their anti-proliferative potential were screened. The aroma profile of the leaves, flowers, and flowers at pre-flowering stages of S. judaica, obtained through SPME was composed of sesquiterpene hydrocarbons (87.7 %, 71.8 %, and 86.2 %, respectively) while the hydro-distilled oil of the dry leaves was rich in oxygenated sesquiterpenes (50.8%). Fresh leaves of S. multicaulis were rich in oxygenated monoterpenes (58.1%), while monoterpene hydrocarbons dominated the blooming flowers (57.2%) and the flowers at the pre-flowering stage (64.7%). The hydro-distilled oil of the dry leaves was rich in oxygenated monoterpenes (77.6%). With doxorubicin as a positive control, no anti-proliferative activity was observed against colorectal cancer cell lines HT29, HCT116, and SW620 using SRB assay for either Salvia spp. In vitro enzymatic starch digestion was evaluated with Acarbose (IC50: 0.2 ± 0.0 µg /mL) as the reference drug. The respective IC50 (mg/mL) values of S. judaica and S. multicaulis aqueous extracts were 4.9 ± 0.4 and 10.3 ± 0.9. Modulation of pancreatic lipase activity (PL) was determined by colorimetry and compared with Orlistat (IC50 : 0.11 ± 0.0 µg/mL). PL-IC50 values (µg/mL) obtained for S. judaica and S. multicaulis were 108.5±6.4 and 31.8 ± 0.8, respectively.

  8. Fiber type conversion by PGC-1α activates lysosomal and autophagosomal biogenesis in both unaffected and Pompe skeletal muscle.

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    Shoichi Takikita

    Full Text Available PGC-1α is a transcriptional co-activator that plays a central role in the regulation of energy metabolism. Our interest in this protein was driven by its ability to promote muscle remodeling. Conversion from fast glycolytic to slow oxidative fibers seemed a promising therapeutic approach in Pompe disease, a severe myopathy caused by deficiency of the lysosomal enzyme acid alpha-glucosidase (GAA which is responsible for the degradation of glycogen. The recently approved enzyme replacement therapy (ERT has only a partial effect in skeletal muscle. In our Pompe mouse model (KO, the poor muscle response is seen in fast but not in slow muscle and is associated with massive accumulation of autophagic debris and ineffective autophagy. In an attempt to turn the therapy-resistant fibers into fibers amenable to therapy, we made transgenic KO mice expressing PGC-1α in muscle (tgKO. The successful switch from fast to slow fibers prevented the formation of autophagic buildup in the converted fibers, but PGC-1α failed to improve the clearance of glycogen by ERT. This outcome is likely explained by an unexpected dramatic increase in muscle glycogen load to levels much closer to those observed in patients, in particular infants, with the disease. We have also found a remarkable rise in the number of lysosomes and autophagosomes in the tgKO compared to the KO. These data point to the role of PGC-1α in muscle glucose metabolism and its possible role as a master regulator for organelle biogenesis - not only for mitochondria but also for lysosomes and autophagosomes. These findings may have implications for therapy of lysosomal diseases and other disorders with altered autophagy.

  9. Inhibitory activity of Iranian plant extracts on growth and biofilm formation by Pseudomonas aeruginosa

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    Mansouri, S.

    2013-01-01

    Full Text Available Aims: Pseudomonas aeruginosa is a drug resistance opportunistic bacterium. Biofilm formation is key factor for survivalof P. aeruginosa in various environments. Polysaccharides may be involved in biofilm formation. The purpose of thisstudy was to evaluate antimicrobial and anti-biofilm activities of seven plant extracts with known alpha-glucosidaseinhibitory activities on different strains of P. aeruginosa.Methodology and results: Plants were extracted with methanol by the maceration method. Antimicrobial activities weredetermined by agar dilution and by growth yield as measured by OD560nm of the Luria Bertani broth (LB culture with orwithout extracts. In agar dilution method, extracts of Quercus infectoria inhibited the growth of all, while Myrtuscommunis extract inhibited the growth of 3 out of 8 bacterial strains with minimum inhibitory concentration (MIC of 1000μg/mL. All extracts significantly (p≤0.003 reduced growth rate of the bacteria in comparison with the control withoutextracts in LB broth at sub-MIC concentrations (500 μg/mL. All plant extracts significantly (p≤0.003 reduced biofilmformation compared to the controls. Glycyrrhiza glabra and Q. infectoria had the highest anti-biofilm activities. Nocorrelation between the alpha-glucosidase inhibitory activity with growth or the intensity of biofilm formation was found.Conclusion, significance and impact of study: Extracts of Q. infectoria and M. communis had the most antimicrobial,while Q. infectoria and G. glabra had the highest anti-biofilm activities. All plant extracts had anti-biofilm activities withmarginal effect on growth, suggesting that the mechanisms of these activities are unrelated to static or cidal effects.Further work to understand the relation between antimicrobial and biofilm formation is needed for development of newmeans to fight the infectious caused by this bacterium in future.

  10. Drug-induced diarrhoea.

    Science.gov (United States)

    Chassany, O; Michaux, A; Bergmann, J F

    2000-01-01

    Diarrhoea is a relatively frequent adverse event, accounting for about 7% of all drug adverse effects. More than 700 drugs have been implicated in causing diarrhoea; those most frequently involved are antimicrobials, laxatives, magnesium-containing antacids, lactose- or sorbitol-containing products, nonsteroidal anti-inflammatory drugs, prostaglandins, colchicine, antineoplastics, antiarrhythmic drugs and cholinergic agents. Certain new drugs are likely to induce diarrhoea because of their pharmacodynamic properties; examples include anthraquinone-related agents, alpha-glucosidase inhibitors, lipase inhibitors and cholinesterase inhibitors. Antimicrobials are responsible for 25% of drug-induced diarrhoea. The disease spectrum of antimicrobial-associated diarrhoea ranges from benign diarrhoea to pseudomembranous colitis. Several pathophysiological mechanisms are involved in drug-induced diarrhoea: osmotic diarrhoea, secretory diarrhoea, shortened transit time, exudative diarrhoea and protein-losing enteropathy, and malabsorption or maldigestion of fat and carbohydrates. Often 2 or more mechanisms are present simultaneously. In clinical practice, 2 major types of diarrhoea are seen: acute diarrhoea, which usually appears during the first few days of treatment, and chronic diarrhoea, lasting more than 3 or 4 weeks and which can appear a long time after the start of drug therapy. Both can be severe and poorly tolerated. In a patient presenting with diarrhoea, the medical history is very important, especially the drug history, as it can suggest a diagnosis of drug-induced diarrhoea and thereby avoid multiple diagnostic tests. The clinical examination should cover severity criteria such as fever, rectal emission of blood and mucus, dehydration and bodyweight loss. Establishing a relationship between drug consumption and diarrhoea or colitis can be difficult when the time elapsed between the start of the drug and the onset of symptoms is long, sometimes up to several

  11. Pompe disease: from pathophysiology to therapy and back again

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    Jeong-A eLim

    2014-07-01

    Full Text Available Pompe disease is a lysosomal storage disorder in which acid alpha-glucosidase is deficient or absent. Deficiency of this lysosomal enzyme results in progressive expansion of glycogen-filled lysosomes in multiple tissues, with cardiac and skeletal muscle being the most severely affected. The clinical spectrum ranges from fatal hypertrophic cardiomyopathy and skeletal muscle myopathy in infants to relatively attenuated forms, which manifest as a progressive myopathy without cardiac involvement. The currently available enzyme replacement therapy proved to be successful in reversing cardiac but not skeletal muscle abnormalities. Although the overall understanding of the disease has progressed, the pathophysiology of muscle damage remains poorly understood. Lysosomal enlargement/rupture has long been considered a mechanism of relentless muscle damage in Pompe disease. In past years, it became clear that this simple view of the pathology is inadequate; the pathological cascade involves dysfunctional autophagy, a major lysosome-dependent intracellular degradative pathway. The autophagic process in Pompe skeletal muscle is affected at the termination stage - impaired autophagosomal-lysosomal fusion. Yet another abnormality in the diseased muscle is the accelerated production of large, unrelated to ageing, lipofuscin deposits - a marker of cellular oxidative damage and a sign of mitochondrial dysfunction. The massive autophagic buildup and lipofuscin inclusions appear to cause a greater effect on muscle architecture than the enlarged lysosomes outside the autophagic regions. Furthermore, the dysfunctional autophagy affects the trafficking of the replacement enzyme and interferes with its delivery to the lysosomes. Several new therapeutic approaches have been tested in Pompe mouse models: substrate reduction therapy, lysosomal exocytosis following the overexpression of transcription factor EB and a closely related but distinct factor E3, and genetic

  12. Action of selected agents on the accumulation of /sup 18/F by Streptococcus mutans

    Energy Technology Data Exchange (ETDEWEB)

    Yotis, W.W.; Zeb, M.; Brennan, P.C.; Kirchner, F.R.; Glendenin, L.E.; Wu-Yuan, C.D.

    1983-01-01

    The action of certain substances known to induce cellular alterations, or encountered in the oral cavity, on the accumulation of /sup 18/F by Streptococcus mutans GS-5 has been investigated. A 62-67% inhibition in the number of /sup 18/F atoms bound per mg dry weight of cells could be induced by a 15 min pretreatment with 2.7 x 10/sup -4/ M cetyltrimethylammoniumbromide, 1 x 10/sup -1/ M acetic anhydride, or 7 x 10/sup -2/ M HCl. Plate counts indicated that alteration of the cellular composition rather than viability was responsible for this diminution in /sup 18/F accumulation. Prior exposure for 15 min of this organism to 1 M HCHO or 0.1 M NaOH did not alter /sup 18/F accumulation. Of the common salts encountered in the oral cavity, CaCl/sub 2/ enhanced /sup 18/F binding. Pretreatment of the assay cells for 15-160 min with 0.1 mg/ml of trypsin, pronase, protease, ..cap alpha..-glucosidase, dextranase, or lactoferrin had no significant effect on the accumulation of /sup 18/. However, pre-exposure of cells for 60 min to 1-10 mg/ml of either amylase or lipase induced a 40-67% inhibition in the binding of /sup 18/F, while lysozyme enhanced the binding of /sup 18/F by the cells. It would appear then that the binding of /sup 18/F by S. mutans may be altered by certain substances encountered in the oral cavity. 17 references, 1 figure, 5 tables.

  13. The effect of methanol extracts of tsao-ko (Amomum tsao-ko Crevost et Lemaire) on digestive enzyme and antioxidant activity in vitro, and plasma lipids and glucose and liver lipids in mice.

    Science.gov (United States)

    Yu, Longquan; Shirai, Nobuya; Suzuki, Hiramitsu; Sugane, Nozomi; Hosono, Tsuyoshi; Nakajima, Yoshijiro; Kajiwara, Masahiro; Takatori, Kazuhiro

    2010-01-01

    Our previous study showed that tsao-ko intake can lower plasma and liver triacylglycerol (TG) concentrations and has hypoglycemic and antioxidant activity in mice. This study involved separating two major fractions (A and B) from the methanol extracts (MeX) of tsao-ko using silica gel column chromatography, and then determining the effect of the fractions in vivo and in vitro to clarify the most effective components of tsao-ko. An intake of MeX and A fraction statistically significantly reduced body lipids and plasma thiobarbitutic acid reactive substances (TBARS) concentrations compared with the control and inhibited lipase and alpha-glucosidase activities. These reductions were not observed in mice fed the B fraction and these inhibitions of B fraction were mild compared with MeX and A fraction. The plasma and liver TG concentrations of each fraction group did not show significant differences compared with the control. The [M-H](+) and maximum UV absorption of the A fraction were 291 m/z and 279 nm, respectively. The peak of A fraction appeared at a similar time to the epicatechin standard in the LC/MS/MS analysis and the MS/MS spectrum of the A fraction was similar to that of the epicatechin standard. It was concluded that the most effective component of tsao-ko for body lipid reduction and hypoglycemic and antioxidant activity was contained in the polar fraction and the evidence suggested that this component could be epicatechin. However, the strongest TG lowering components of tsao-ko may be methanol insoluble.

  14. Type 2 Diabetes Mellitus: A Review of Current Trends

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    Abdulfatai B. Olokoba

    2012-07-01

    Full Text Available Type 2 diabetes mellitus (DM is a chronic metabolic disorder in which prevalence has been increasing steadily all over the world. As a result of this trend, it is fast becoming an epidemic in some countries of the world with the number of people affected expected to double in the next decade due to increase in ageing population, thereby adding to the already existing burden for healthcare providers, especially in poorly developed countries. This review is based on a search of Medline, the Cochrane Database of Systemic Reviews, and citation lists of relevant publications. Subject heading and key words used include type 2 diabetes mellitus, prevalence, current diagnosis, and current treatment. Only articles in English were included. Screening and diagnosis is still based on World Health Organization (WHO and American Diabetes Association (ADA criteria which include both clinical and laboratory parameters. No cure has yet been found for the disease; however, treatment modalities include lifestyle modifications, treatment of obesity, oral hypoglycemic agents, and insulin sensitizers like metformin, a biguanide that reduces insulin resistance, is still the recommended first line medication especially for obese patients. Other effective medications include non-sulfonylurea secretagogues, thiazolidinediones, alpha glucosidase inhibitors, and insulin. Recent research into the pathophysiology of type 2 DM has led to the introduction of new medications like glucagon-like peptide 1 analogoues: dipeptidyl peptidase-IV inhibitors, inhibitors of the sodium-glucose cotransporter 2 and 11ß-hydroxysteroid dehydrogenase 1, insulin-releasing glucokinase activators and pancreatic-G-protein-coupled fatty-acid-receptor agonists, glucagon-receptor antagonists, metabolic inhibitors of hepatic glucose output and quick-release bromocriptine. Inhaled insulin was licensed for use in 2006 but has been withdrawn from the market because of low patronage.

  15. To study the antihyperglycaemic and lipid lowering effect of garlic as an adjunct to metformin in patients of type 2 diabetes mellitus with obesity

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    Simran Chhatwal

    2012-02-01

    Full Text Available Background: Diabetes mellitus is a metabolic disorder of multiple aetiology characterized by chronic hyperglycaemia associated with disturbances of carbohydrate, fat and protein metabolism. The treatment constitutes lifestyle management, exercise, weight control and antihyperglycaemic drugs like sulfonylureas, biguanides, alpha-glucosidase inhibitors, thiazolidinediones, and meglitinide. Garlic has shown to have anti-hyperglycaemic and lipid lowering effects in various animal and human studies. Thus, this study was conducted to assess the antihyperglycaemic and lipid-lowering properties of Garlic in type2 diabetes patients with obesity. Methods: This was an open labelled prospective comparative study conducted on Type 2 diabetes mellitus patients with obesity where a total of 60 patients divided into two groups of 30 each (of either sex were enrolled. Group 1 was given Tab. metformin 500mg BD/TDS after meals. Group 2 was given Tab. metformin in a dose of 500mg BD/TDS after meals along with Cap. Garlic (Allium sativum 250mg BD. Patients were routinely investigated for fasting blood sugar, HbA1c and lipid profile i.e. Serum Cholesterol, HDL-C, Triglycerides and LDL-C at the start of the study. Patients were followed up at an interval of two weeks upto 12 weeks. Data obtained at the end of the study was statistically analysed using Student's- t test. Results: It was observed that both metformin and garlic reduced FBG and HbA1c significantly but percentage reduction in FBG was more with garlic but, change in HbA1c was not significant. Fall in total CHL, TG, LDL and an increase in HDL were more pronounced in patients treated with Garlic when given along with Metformin. Conclusions: Therefore, garlic showed better results as an antihyperglycaemic and lipid lowering agent. [Int J Basic Clin Pharmacol 2012; 1(1.000: 22-26

  16. Hydrolytic and ligninolytic enzyme activities in the Pb contaminated soil inoculated with litter-decomposing fungi.

    Science.gov (United States)

    Kähkönen, Mika A; Lankinen, Pauliina; Hatakka, Annele

    2008-06-01

    The impact of Pb contamination was tested to five hydrolytic (beta-glucosidase, beta-xylosidase, beta-cellobiosidase, alpha-glucosidase and sulphatase) and two ligninolytic (manganese peroxidase, MnP and laccase) enzyme activities in the humus layer in the forest soil. The ability of eight selected litter-degrading fungi to grow and produce extracellular enzymes in the heavily Pb (40 g Pb of kg ww soil(-1)) contaminated and non-contaminated soil in the non-sterile conditions was also studied. The Pb content in the test soil was close to that of the shooting range at Hälvälä (37 g Pb of kg ww soil(-1)) in Southern Finland. The fungi were Agaricus bisporus, Agrocybe praecox, Gymnopus peronatus, Gymnopilus sapineus, Mycena galericulata, Gymnopilus luteofolius, Stropharia aeruginosa and Stropharia rugosoannulata. The Pb contamination (40 g Pb of kg ww soil(-1)) was deleterious to all five studied hydrolytic enzyme activities after five weeks of incubation. All five hydrolytic enzyme activities were significantly higher in the soil than in the extract of the soil indicating that a considerable part of enzymes were particle bound in the soils. Hydrolytic enzyme activities were higher in the non-contaminated soil than in the Pb contaminated soil. Fungal inocula increased the hydrolytic enzyme activities beta-cellobiosidase and beta-glucosidase in non-contaminated soils. All five hydrolytic enzyme activities were similar with fungi and without fungi in the Pb contaminated soil. This was in line that Pb contamination (40 g Pb of kg ww soil(-1)) depressed the growth of all fungi compared to those grown without Pb in the soil. Laccase and MnP activities were low in both Pb contaminated and non-contaminated soil cultures. MnP activities were higher in soil cultures containing Pb than without Pb. Our results showed that Pb in the shooting ranges decreased fungal growth and microbial functioning in the soil.

  17. Ten years of dengue drug discovery: progress and prospects.

    Science.gov (United States)

    Lim, Siew Pheng; Wang, Qing-Yin; Noble, Christian G; Chen, Yen-Liang; Dong, Hongping; Zou, Bin; Yokokawa, Fumiaki; Nilar, Shahul; Smith, Paul; Beer, David; Lescar, Julien; Shi, Pei-Yong

    2013-11-01

    To combat neglected diseases, the Novartis Institute of Tropical Diseases (NITD) was founded in 2002 through private-public funding from Novartis and the Singapore Economic Development Board. One of NITD's missions is to develop antivirals for dengue virus (DENV), the most prevalent mosquito-borne viral pathogen. Neither vaccine nor antiviral is currently available for DENV. Here we review the progress in dengue drug discovery made at NITD as well as the major discoveries made by academia and other companies. Four strategies have been pursued to identify inhibitors of DENV through targeting both viral and host proteins: (i) HTS (high-throughput screening) using virus replication assays; (ii) HTS using viral enzyme assays; (iii) structure-based in silico docking and rational design; (iv) repurposing hepatitis C virus inhibitors for DENV. Along the developmental process from hit finding to clinical candidate, many inhibitors did not advance beyond the stage of hit-to-lead optimization, due to their poor selectivity, physiochemical or pharmacokinetic properties. Only a few compounds showed efficacy in the AG129 DENV mouse model. Two nucleoside analogs, NITD-008 and Balapiravir, entered preclinical animal safety study and clinic trial, but both were terminated due to toxicity and lack of potency, respectively. Celgosivir, a host alpha-glucosidase inhibitor, is currently under clinical trial; its clinical efficacy remains to be determined. The knowledge accumulated during the past decade has provided a better rationale for ongoing dengue drug discovery. Though challenging, we are optimistic that this continuous, concerted effort will lead to an effective dengue therapy.

  18. Impact of glucose-lowering agents on the risk of cancer in type 2 diabetic patients. The Barcelona case-control study.

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    Rafael Simó

    Full Text Available BACKGROUND: The aim of the present study is to evaluate the impact of glucose-lowering agents in the risk of cancer in a large type 2 diabetic population. METHODS: A nested case-control study was conducted within a defined cohort (275,164 type 2 diabetic patients attending 16 Primary Health Care Centers of Barcelona. Cases (n = 1,040 comprised those subjects with any cancer diagnosed between 2008 and 2010, registered at the Cancer Registry of Hospital Vall d'Hebron (Barcelona. Three control subjects for each case (n = 3,120 were matched by age, sex, diabetes duration, and geographical area. The treatments analyzed (within 3 years prior to cancer diagnosis were: insulin glargine, insulin detemir, human insulin, fast-acting insulin and analogues, metformin, sulfonylureas, repaglinide, thiazolidinediones, dipeptidyl peptidase-4 inhibitors, and alpha glucosidase inhibitors. Conditional logistic regressions were used to calculate the risk of cancer associated with the use of each drug adjusted by age, BMI, dose and duration of treatment, alcohol use, smoking habit, and diabetes duration. RESULTS: No differences were observed between case and control subjects for the proportion, dose or duration of exposure to each treatment. None of the types of insulin and oral agents analyzed showed a significant increase in the risk of cancer. Moreover, no cancer risk was observed when glargine was used alone or in combination with metformin. CONCLUSIONS: Our results suggest that diabetes treatment does not influence the risk of cancer associated with type 2 diabetes. Therefore, an eventual increase of cancer should not be a reason for biasing the selection of any glucose-lowering treatment in type 2 diabetic population.

  19. A novel RNA binding protein that interacts with NMDA R1 mRNA: regulation by ethanol.

    Science.gov (United States)

    Anji, Antje; Kumari, Meena

    2006-05-01

    Excitatory NMDA receptors are an important target of ethanol. Chronic ethanol exposure, in vivo and in vitro, increases polypeptide levels of NR1 subunit, the key subunit of functional NMDA receptors. In vitro, chronic ethanol treatment increases the half-life of NR1 mRNA and this observation is dependent on new protein synthesis. The present study was undertaken to locate cis-acting region(s) within the NR1 3'-untranslated region (UTR) and identify NR1 3'-UTR binding trans-acting proteins expressed in mouse fetal cortical neurons. Utilizing RNA gel shift assays we identified a 156-nt cis-acting region that binds to polysomal trans-acting proteins. This binding was highly specific as inclusion of cyclophilin RNA or tRNA did not interfere with cis-trans interactions. Importantly, the 3'-UTR binding activity was significantly up-regulated in the presence of ethanol. UV cross-link analysis detected three NR1 3'-UTR binding proteins and their molecular mass calculated by Northwestern analysis was approximately 88, 60 and 47 kDa, respectively. Northwestern analysis showed a significant up-regulation of the 88-kDa protein after chronic ethanol treatment. The 88-kDa protein was purified and identified by tandem mass spectrometry as the beta subunit of alpha glucosidase II (GIIbeta). That GIIbeta is indeed a trans-acting protein and binds specifically to 3'-UTR of NR1 mRNA was confirmed by RNA gel mobility supershift assays and immuno RT-PCR. Western blotting data established a significant increase of GIIbeta polypeptide in chronic ethanol-exposed fetal cortical neurons. We hypothesize that the identified cis-acting region and the associated RNA-binding proteins are important regulators of NR1 subunit gene expression.

  20. Changing Patterns of Glucose-Lowering Medication Use in VA Nursing Home Residents with Diabetes, 2005 – 2011

    Science.gov (United States)

    Lee, Sei J.; Stijacic-Cenzer, Irena; Barnhart, Caroline; McClymont, Keelan; Steinman, Michael A

    2015-01-01

    Objective Although nursing home (NH) residents make up a large and growing proportion of Americans with diabetes mellitus, little is known about how glucose-lowering medications are used in this population. We sought to examine glucose-lowering medication use in Veterans Affairs (VA) NH residents with diabetes between 2005–2011. Research Design and Methods Retrospective cohort study, utilizing linked laboratory, pharmacy, administrative and NH Minimum Dataset (MDS) 2.0 databases in 123 VA NHs. A total of 9,431 long-stay (>90 days) VA NH residents over age 65 followed for 52,313 person-quarters. We identified receipt of glucose lowering medications including insulin, metformin, sulfonylureas, thiazolidinediones and others (alpha-glucosidase inhibitors, meglitinides, glucagon like peptide-1 analogs, dipeptidyl peptidase-4 inhibitors and amylin analogues) per quarter. Results The rates of sulfonylurea use in long-stay NH residents dropped dramatically from 24% in 2005 to 12% in 2011 (p<0.001), driven in large part by the dramatic decrease in glyburide use (10% to 2%, p<0.001). There was sharp drop in thiazolidinedione use in 2007 (4% to <1%, p<0.001). Metformin use was stable, ranging between 7% to 9% (p=0.24). Insulin use increased slightly from 30% to 32% (p<0.001). Use of other classes of glucose-lowering medications was stable (p=0.22) and low, remaining below 1.3%. Conclusions and Relevance Between 2005 and 2011, there were dramatic declines in use of sulfonylureas and thiazolidinediones in VA NH residents, suggesting that prescribing practices can be quickly changed in this setting. PMID:26272298

  1. Cardiovascular disease in the JCR:LA-cp rat.

    Science.gov (United States)

    Russell, J C; Graham, S E; Richardson, M

    1998-11-01

    The JCR:LA-cp rat is one of a number of strains that carry the mutant autosomal recessive cp gene. Animals, of all strains, that are homozygous, for the gene (cp/cp) become obese, insulin resistant, and hypertriglyceridemic. Heterozygotes or homozygous normal rats (+/+) are lean and metabolically normal. The JCR:LA-cp rat is unique in the development of a frank vasculopathy with atherosclerotic lesions and associated ischemic myocardial lesions. The cardiovascular disease is strongly correlated with the hyperinsulinemia, which develops as the animals mature from 4 to 8 weeks of age. The hyperinsulinemia can be decreased by marked food restriction, ethanol consumption, or reduction of the postprandial glucose and insulin responses through the use of alpha-glucosidase inhibitors. Any treatment that reduces plasma insulin levels is associated with a reduction in cardiovascular disease. In contrast, a reduction in plasma triglyceride concentrations, alone, has no effect on end-stage lesions. JCR:LA-cp rats, particularly those that are cp/cp, are, however, sensitive to cholesterol in the diet, unlike other strains that are highly resistant. Further, the rats have abnormal vascular smooth muscle cells that, especially in the cp/cp animals, are hyperplastic and activated and migrate into the intimal space. Our findings suggest that susceptibility to cardiovascular disease requires hypermsulinemic stress coupled with excessive dietary intake and the presence of one or more other necessary, but not sufficient, genetic factors. One of these may be a genetic abnormality of vascular smooth muscle cells. A similar situation may occur in humans.

  2. Inhibitory mechanism of acarbose and 1-deoxynojirimycin derivatives on carbohydrases in rat small intestine.

    Science.gov (United States)

    Samulitis, B K; Goda, T; Lee, S M; Koldovský, O

    1987-01-01

    The inhibitory action and mechanism of inhibition of two types of alpha-glucosidase inhibitors, acarbose (Bay-g-5421) and 1-deoxynojirimycin derivatives (Bay-m-1099 and Bay-o-1248), on small intestinal carbohydrases (sucrase, isomaltase, glucoamylase, trehalase and lactase) and pancreatic alpha-amylase were compared in vitro using small intestinal brush border membranes and pancreatic homogenates from adult Sprague-Dawley rats. Acarbose at a low (4 microM) concentration strongly inhibited the activities of glucoamylase, alpha-amylase and sucrase (98, 68, and 63%, respectively). At a high (200 microM) concentration, isomaltase activity was also inhibited (28%); effects on trehalase and lactase activities were negligible. Both the 1-deoxynojirimycin derivatives were even more potent inhibitors of sucrase (Ki = 8.6 x 10(-8) M for Bay-m-1099;Ki = 5.0 X 10(-8) M for Bay-o-1248) than acarbose (Ki = 9.9 x 10(-7) M). Whereas glucoamylase activity was strongly inhibited by the 1-deoxynojirimycin derivatives, alpha-amylase activity was not. In contrast to acarbose, the 1-deoxynojirimycin derivatives at high concentrations (20-200 microM) inhibited considerably trehalase and lactase (a beta-galactosidase) activities. The inhibition of lactase activity was stronger by Bay-m-1099 (Ki = 4.9 X 10(-6) M) than by Bay-o-1248 (Ki = 6.7 X 10(-5) M). Where inhibition was seen, kinetic analysis showed fully competitive inhibition of sucrase, isomaltase, trehalase, glucoamylase and lactase by all three inhibitors.

  3. Expression and characterization of α-glucosidase Ⅲ in the dwarf honeybee, Apis florea (Hymenoptera: Apoidea:Apidae)

    Institute of Scientific and Technical Information of China (English)

    CHANPEN CHANCHAO; RUMPALAI PADOONGSUPALAI; POLKIT SANGVANICH

    2007-01-01

    Alpha-glucosidase is synthesized in the hypopharyngeal glands located in the head of worker bees including Apis florea. To analyze the developmental stage-specific expression of the α-glucosidase gene in A. florea, total RNA was isolated from eggs, and the heads of nurse and forager bees. By reverse transcription polymerase chain reaction (RTPCR), it was shown that the highest expression levels of the α-glucosidase Ⅲ gene, in the three examined developmental stadia, were found in forager bees, with much lower expression levels in nurse bees and no detectable expression in eggs. A complete α-glucosidase Ⅲ cDNA was obtained by RT-PCR and sequenced. The 1 701 bp cDNA nucleotide sequence and the predicted 567 amino acids it encodes were assayed by BLASTn,BLASTp and BLASTx programs and revealed a 95% and 94% similarity to the A. mellifera α-glucosidase Ⅲ gene at the DNA and amino acid sequence levels, respectively. For purification of the active encoded enzyme, forager bee heads were homogenized in sodium phosphate buffer solution and the crude extract (0.30 U/mg) sequentially precipitated with 95% saturated ammonium sulfate (0.18 U/mg), and purified by DEAE cellulose ion exchange chromatography (0.17 U/mg), and gel filtration on Superdex 200 (0.52 U/mg).After resolution through sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a single enzymically active band (73 kDa) was identified from renatured substrate gels.Excision of this band, elution of the protein and tryptic peptide digestives identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) revealed six matching masses to the A. mellifera (Q17958) and predicted A. florea α-glucosidase Ⅲ protein with 12% coverage, supporting the probable purification of the same α-glucosidase Ⅲ protein as that encoded by the cloned cDNA.

  4. Contribution of mucosal maltase-glucoamylase activities to mouse small intestinal starch alpha-glucogenesis.

    Science.gov (United States)

    Quezada-Calvillo, Roberto; Robayo-Torres, Claudia C; Opekun, Antone R; Sen, Partha; Ao, Zihua; Hamaker, Bruce R; Quaroni, Andrea; Brayer, Gary D; Wattler, Sigrid; Nehls, Michael C; Sterchi, Erwin E; Nichols, Buford L

    2007-07-01

    Digestion of starch requires activities provided by 6 interactive small intestinal enzymes. Two of these are luminal endo-glucosidases named alpha-amylases. Four are exo-glucosidases bound to the luminal surface of enterocytes. These mucosal activities were identified as 4 different maltases. Two maltase activities were associated with sucrase-isomaltase. Two remaining maltases, lacking other identifying activities, were named maltase-glucoamylase. These 4 activities are better described as alpha-glucosidases because they digest all linear starch oligosaccharides to glucose. Because confusion persists about the relative roles of these 6 enzymes, we ablated maltase-glucoamylase gene expression by homologous recombination in Sv/129 mice. We assayed the alpha-glucogenic activities of the jejunal mucosa with and without added recombinant pancreatic alpha-amylase, using a range of food starch substrates. Compared with wild-type mucosa, null mucosa or alpha-amylase alone had little alpha-glucogenic activity. alpha-Amylase amplified wild-type and null mucosal alpha-glucogenesis. alpha-Amylase amplification was most potent against amylose and model resistant starches but was inactive against its final product limit-dextrin and its constituent glucosides. Both sucrase-isomaltase and maltase-glucoamylase were active with limit-dextrin substrate. These mucosal assays were corroborated by a 13C-limit-dextrin breath test. In conclusion, the global effect of maltase-glucoamylase ablation was a slowing of rates of mucosal alpha-glucogenesis. Maltase-glucoamylase determined rates of digestion of starch in normal mice and alpha-amylase served as an amplifier for mucosal starch digestion. Acarbose inhibition was most potent against maltase-glucoamylase activities of the wild-type mouse. The consortium of 6 interactive enzymes appears to be a mechanism for adaptation of alpha-glucogenesis to a wide range of food starches.

  5. Luminal starch substrate "brake" on maltase-glucoamylase activity is located within the glucoamylase subunit.

    Science.gov (United States)

    Quezada-Calvillo, Roberto; Sim, Lyann; Ao, Zihua; Hamaker, Bruce R; Quaroni, Andrea; Brayer, Gary D; Sterchi, Erwin E; Robayo-Torres, Claudia C; Rose, David R; Nichols, Buford L

    2008-04-01

    The detailed mechanistic aspects for the final starch digestion process leading to effective alpha-glucogenesis by the 2 mucosal alpha-glucosidases, human sucrase-isomaltase complex (SI) and human maltase-glucoamylase (MGAM), are poorly understood. This is due to the structural complexity and vast variety of starches and their intermediate digestion products, the poorly understood enzyme-substrate interactions occurring during the digestive process, and the limited knowledge of the structure-function properties of SI and MGAM. Here we analyzed the basic catalytic properties of the N-terminal subunit of MGAM (ntMGAM) on the hydrolysis of glucan substrates and compared it with those of human native MGAM isolated by immunochemical methods. In relation to native MGAM, ntMGAM displayed slower activity against maltose to maltopentose (G5) series glucose oligomers, as well as maltodextrins and alpha-limit dextrins, and failed to show the strong substrate inhibitory "brake" effect caused by maltotriose, maltotetrose, and G5 on the native enzyme. In addition, the inhibitory constant for acarbose was 2 orders of magnitude higher for ntMGAM than for native MGAM, suggesting lower affinity and/or fewer binding configurations of the active site in the recombinant enzyme. The results strongly suggested that the C-terminal subunit of MGAM has a greater catalytic efficiency due to a higher affinity for glucan substrates and larger number of binding configurations to its active site. Our results show for the first time, to our knowledge, that the C-terminal subunit of MGAM is responsible for the MGAM peptide's "glucoamylase" activity and is the location of the substrate inhibitory brake. In contrast, the membrane-bound ntMGAM subunit contains the poorly inhibitable "maltase" activity of the internally duplicated enzyme.

  6. HYPERTROPHIC OBSTRUCTIVE CARDIOMYOPATHY IN AN 8-MONTH OLD FEMALE INFANT SUSPECTED INFANTILE ONSET POMPE DISEASE

    Directory of Open Access Journals (Sweden)

    Made Dwi Purnami

    2015-05-01

    Full Text Available Hypertrophic cardiomyopathy (HCM is an autosomal dominant cardiac disorder marked with muscular hypertrophy of the left ventricle, associated obstruction of left ventricular outflow. About 0.2% of all cases worldwide. The majority of patients are asymptomatic, and some present with severe activity- limiting symptoms. The diagnosis of HCM before the age of 2 years is rare and usually discovered by chance, during the investigation of a murmur. Progressive disease characterized by prominent cardiomegaly, cadiomyopathy, hepatomegaly, musle weakness or hypotonia, respiratory distress, feeding difficulties and failure to thrive as presenting sign and symptoms are often referred to infantile Pompe disease. A deficiency of of the enzyme acid alpha glucosidase disease, result in lysosomal accumulation of glycogen in heart and skeletal muscle. Cardiorespiratory failure is the cause of significant morbidity and mortality in the first year of life. We reported a rare case, 8 month-old female with frequent respiratory distress since 2 months before admission. Physical examination showed dyspnea with chest wall retraction, no cyanosis, with grade III systolic murmur at midclavicular line sinistra, ICS IV- V and floopy infant. Chest films showed   pneumonia and cardiomegaly. The echocardiogram demonstrated bi-ventricular and interventricular hypertrophy with left ventricular obstruction. Laboratory finding there was increased levels of glutamic oxaloacetic acid transferase, alanin aminotransferase, and lactate dehydrogenase. Patient was diagnosed with hypertrophic obstructive cardiomyopathy of suspected infantile onset pompe disease. Despite medical treatment with propanolol dan diuretics, there was no significant improvement and she was died after 26th days of treatment in intermediate ward. [MEDICINA 2014;45:108-14]    

  7. Treponema maltophilum sp. nov., a small oral spirochete isolated from human periodontal lesions.

    Science.gov (United States)

    Wyss, C; Choi, B K; Schüpbach, P; Guggenheim, B; Göbel, U B

    1996-07-01

    A novel culture medium for cultivation of fastidious oral anaerobes is described. This medium, OMIZ-Pat, consists of a rich chemically defined basal medium supplemented with asialofetuin, as well as yeast extract and Neopeptone fractions. Addition of 1 mg of rifampin per liter and 100 mg of fosfomycin per liter allowed routine isolation of spirochetes by a limit dilution method in 96-well plates containing liquid OMIZ-Pat. In addition to members of the four previously recognized species of oral treponemes (Treponema denticola, Treponema pectinovorum, Treponema socranskii, and Treponema vincentii), 26 previously undescribed spirochete strains belonging to one group were isolated. We propose the name Treponema maltophilum sp. nov. for these small spirochetes, which have two endoflagella; one endoflagellum is attached at each cell pole, and the endoflagella overlap in the middle of the cell. Growth of these organisms was dependent on a carbohydrate like D-arabinose, L-fucose, D-maltose, L-rhamnose, D-ribose, D-sucrose, or D-trehalose and was inhibited by fetal bovine serum. T. Maltophilum is distinguished from other oral Treponema species by its 16S rRNA sequence, its protein and antigen patterns as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting, and its characteristic alpha-glucosidase activity. The strains included in the new species on the basis of their 16S rRNA sequences are heterogeneous with respect to their alpha-fucosidase, and beta-glucuronidase activities, their dependence on N-acetylglucosamine, and their antigens as detected with patient antibodies. Strain BR is designated the type strain, and strains HO2A and PNA1 are reference strains of the new species.

  8. Bacterial and enzymatic bioassays for toxicity testing in the environment.

    Science.gov (United States)

    Bitton, G; Koopman, B

    1992-01-01

    Microbioassays using bacteria or enzymes are increasingly applied to measure chemical toxicity in the environment. Attractive features of these assays may include low cost, rapid response to toxicants, high sample throughput, modest laboratory equipment and space requirements, low sample volume, portability, and reproducible responses. Enzymatic tests rely on measurement of either enzyme activity or enzyme biosynthesis. Dehydrogenases are the enzymes most used in toxicity testing. Assay of dehydrogenase activity is conveniently carried out using oxidoreduction dyes such as tetrazolium salts. Other enzyme activity tests utilize ATPases, esterases, phosphatases, urease, luciferase, beta-galactosidase, protease, amylase, or beta-glucosidase. Recently, the inhibition of enzyme (beta-galactosidase, tryptophanase, alpha-glucosidase) biosynthesis has been explored as a basis for toxicity testing. Enzyme biosynthesis was found to be generally more sensitive to organic chemicals than enzyme activity. Bacterial toxicity tests are based on bioluminescence, motility, growth, viability, ATP, oxygen uptake, nitrification, or heat production. An important aspect of bacterial tests is the permeability of cells to environmental toxicants, particularly organic chemicals of hydrophobic nature. Physical, chemical, and genetic alterations of the outer membrane of E. coli have been found to affect test sensitivity to organic toxicants. Several microbioassays are now commercially available. The names of the assays and their basis are: Microtox (bioluminescence), Polytox (respiration), ECHA Biocide Monitor (dehydrogenase activity), Toxi-Chromotest (enzyme biosynthesis), and MetPAD (enzyme activity). An important feature common to these tests is the provision of standardized cultures of bacteria in freeze-dried form. Two of the more recent applications of microbioassays are in sediment toxicity testing and toxicity reduction evaluation. Sediment pore water may be assayed directly or

  9. Antidiabetic treatment with gliptins: focus on cardiovascular effects and outcomes.

    Science.gov (United States)

    Fisman, Enrique Z; Tenenbaum, Alexander

    2015-01-01

    The traditional oral pharmacological therapy for type 2 diabetes mellitus (T2DM) has been based on the prescription of metformin, a biguanide, as first line antihyperglycemic agent world over. It has been demonstrated that after 3 years of treatment, approximately 50% of diabetic patients could achieve acceptable glucose levels with monotherapy; but by 9 years this had declined to only 25%. Therefore, the implementation of a combined pharmacological therapy acting via different pathways becomes necessary, and its combination with a compound of the sulfonylurea group was along decades the most frequently employed prescription in routine clinical practice. Meglitinides, glitazones and alpha-glucosidase inhibitors were subsequently developed, but the five mentioned groups of oral antihyperglycemic agents are associated with variable degrees of undesirable or even severe cardiovascular events. The gliptins-also called dipeptidyl peptidase 4 (DPP4) inhibitors--are an additional group of antidiabetic compounds with increasing clinical use. We review the status of the gliptins with emphasis on their capabilities to positively or negatively affect the cardiovascular system, and their potential involvement in major adverse cardiovascular events (MACE). Alogliptin, anagliptin, linagliptin, saxagliptin, sitagliptin, teneligliptin and vildagliptin are the compounds currently in clinical use. Regardless differences in chemical structure and metabolic pathways, gliptins as a group exert favorable changes in experimental models. These changes, as an almost general rule, include improved endothelial function, reduction of inflammatory markers, oxidative stress ischemia/reperfusion injury and atherogenesis. In addition, increased adiponectin levels and modest decreases in lipidemia and blood pressure were reported. In clinical settings, several trials--notably the longer one, employing sitagliptin, with a mean follow-up period of 3 years--did not show an increased risk for ischemic

  10. Acarbose improves glycemic control and reduces body weight: Subanalysis data of South Asia region

    Directory of Open Access Journals (Sweden)

    S Kalra

    2013-01-01

    Full Text Available Alpha-glucosidase inhibitors (AGIs are widely used especially in Asian countries as a treatment option for type 2 diabetes patients with high postprandial glycaemia. However, data from South Asia region is very limited. In order to examine the effect of AGI in real-life setting, 10 PMS/NIS from all over the world from the launch of acarbose to date were pooled in one database and exploratory analysis was performed for glycemic parameters and weight. In total 62,905 patients were pooled from 21 countries and regions. Mean follow up (± SD was 12.2 ± 4.8 weeks (range 0.1-108.9. From South Asia region (India and Pakistan, 8,738 Asian patients were enrolled. Mean PPG decreased from 240.0 and 261.1 mg/dl at baseline by 70.26 ± 65.10 and 82.96 ± 56.59 mg/dl at the last visit in total and South Asian populations, respectively (n = 53,883; n = 7,991, P < 0.0001 for both. Mean FPG decreased from 171.6 and 176.5 mg/dl at baseline by 38.48 ± 47.83 and 49.59 ± 41.41 mg/dl at the last visit in total and South Asian populations, respectively (n = 56,672; n = 7,837, P < 0.0001 for both. Mean HbA1c decreased from 8.4 and 8.4% at baseline by 1.11 ± 1.31% and 0.91 ± 0.93% at the last visit in total and South Asian populations, respectively (n = 38,843; n = 2,343, P < 0.0001 for both. Mean relative reduction of body weight (BW was 1.40 ± 3.28% and 1.10 ± 3.39% at the last visit for mean baseline BW 73.6 and 74.2 kg in total and South Asian populations, respectively (n = 54,760; n = 7,718, P < 0.0001 for both. Consistent with RCT meta-analyses, post-hoc analysis of real-life data showed acarbose treatment improved glycaemic control and reduced the BW. Acarbose treatment in real life setting showed significant reductions in all glycemic parameters and BW in Asian patients from South Asia region.

  11. Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs

    Science.gov (United States)

    Ali, Muhammad Yousaf; Ashraf, Muhammad Aleem; Mansoor, Shahid; Shahid, Ahmad Ali; Brown, Judith K.

    2017-01-01

    Control of the whitefly Bemisia tabaci (Genn.) agricultural pest and plant virus vector relies on the use of chemical insecticides. RNA-interference (RNAi) is a homology-dependent innate immune response in eukaryotes, including insects, which results in degradation of the corresponding transcript following its recognition by a double-stranded RNA (dsRNA) that shares 100% sequence homology. In this study, six whitefly ‘gut’ genes were selected from an in silico-annotated transcriptome library constructed from the whitefly alimentary canal or ‘gut’ of the B biotype of B. tabaci, and tested for knock down efficacy, post-ingestion of dsRNAs that share 100% sequence homology to each respective gene target. Candidate genes were: Acetylcholine receptor subunit α, Alpha glucosidase 1, Aquaporin 1, Heat shock protein 70, Trehalase1, and Trehalose transporter1. The efficacy of RNAi knock down was further tested in a gene-specific functional bioassay, and mortality was recorded in 24 hr intervals, six days, post-treatment. Based on qPCR analysis, all six genes tested showed significantly reduced gene expression. Moderate-to-high whitefly mortality was associated with the down-regulation of osmoregulation, sugar metabolism and sugar transport-associated genes, demonstrating that whitefly survivability was linked with RNAi results. Silenced Acetylcholine receptor subunit α and Heat shock protein 70 genes showed an initial low whitefly mortality, however, following insecticide or high temperature treatments, respectively, significantly increased knockdown efficacy and death was observed, indicating enhanced post-knockdown sensitivity perhaps related to systemic silencing. The oral delivery of gut-specific dsRNAs, when combined with qPCR analysis of gene expression and a corresponding gene-specific bioassay that relates knockdown and mortality, offers a viable approach for functional genomics analysis and the discovery of prospective dsRNA biopesticide targets. The

  12. Carbon isotope labeling in boreal forests to assess roles of fungal species in decomposition

    Science.gov (United States)

    Treseder, K. K.; Czimczik, C. I.; Trumbore, S. E.; Allison, S. D.

    2006-12-01

    by saprotrophic fungi, which may be due to an absence or lack of activity of starch-degrading fungal species during the study period. Potential activity of the starch-metabolizing enzyme alpha-glucosidase was only 0.59 +/- 0.17 ìmol h-1 g dry soil-1, which was 7 times less than activity of beta-glucosidase, which breaks down cellulose. The slow turnover of lignocellulose-C was consistent with slow decomposition rates of plant litter in this biome.

  13. Teneligliptin real-world efficacy assessment of type 2 diabetes mellitus patients in India (TREAT-INDIA study

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    Ghosh S

    2016-11-01

    Full Text Available Sujoy Ghosh,1 Shailesh Trivedi,2 Debmalya Sanyal,3 KD Modi,4 Sandeep Kharb5 1Department of Endocrinology, The Institute of Post Graduate Medical Education and Research (IPGMER, Kolkata, West Bengal, 2Anand Hospital, Vadodara, Gujarat, 3Department of Endocrinology, KPC Medical College, Kolkata, West Bengal, 4Dr Modi’s Clinic (DMC, Department of Endocrinology at Medwin Hospital, Hyderabad, Telangana, 5ASIAN Hospital, Faridabad, Haryana, India Background and aims: Teneligliptin was introduced in India in May 2015. It has gained popularity and is already widely prescribed in type 2 diabetes mellitus (T2DM. This “real life” data collection was conducted to assess the efficacy of teneligliptin in Indian T2DM patients. Methods: Predesigned structured proforma was used to collect information from the prescribing physicians regarding the efficacy of teneligliptin when prescribed as monotherapy as well as combination therapy with other antidiabetic drugs in T2DM patients. Information on the glycemic parameters at baseline prior to starting teneligliptin and at the end of 3 months therapy was collected. The efficacy was assessed by analyzing the mean change in 3-month values of glycosylated hemoglobin (HbA1c, fasting plasma glucose (FPG, and postprandial plasma glucose (PPG.Results: Data of 4305 patients was available for analysis. There was statistically significant improvement in mean HbA1c, FPG, and PPG with teneligliptin therapy. Means changes in HbA1c, FPG, and PPG were −1.37%±1.15%, 51.29±35.41 mg/dL, and 80.89±54.27 mg/dL, respectively. Subgroup analysis revealed that HbA1c (% reduction with teneligliptin when used as monotherapy, add-on to metformin or add-on to metformin plus sulfonylureas combination, add-on to metformin plus alpha glucosidase inhibitor combination or add-on to insulin was 0.98±0.53, 1.07±0.83, 1.46±1.33, 1.43±0.80, and 1.55±1.05, respectively.Conclusion: Real-world data suggests that teneligliptin significantly

  14. Recent developments and emerging therapies for type 2 diabetes mellitus.

    Science.gov (United States)

    Evans, A J; Krentz, A J

    1999-08-01

    Most patients with type 2 (non-insulin-dependent) diabetes mellitus require pharmacotherapy, initially as monotherapy and subsequently in combination, as adjuncts to diet and exercise. Exogenous insulin is ultimately required in a substantial proportion, reflecting the progressive natural history of the disease. Sulphonylureas and biguanides have been employed for over 4 decades as oral antidiabetic agents, but they have a limited capacity to provide long term glycaemic control and can cause serious adverse effects. Thus, more efficacious and tolerable antidiabetic agents are required. Recent years have witnessed the introduction of agents with novel modes of action, that is, the alpha-glucosidase inhibitors acarbose and miglitol (which reduce postprandial hyperglycaemia) and the first of the thiazolidinedione insulinsensitising drugs--troglitazone and rosiglitazone. Although the former has been withdrawn in some countries due to adverse effects, another 'glitazone' pioglitazone is expected to be approved in the near future. Other recently introduced drugs include glimepiride and the meglitinide insulin secretagogue, repaglinide. Attention is also focusing increasingly on combination therapy using insulin together with sulphonylureas, metformin or troglitazone. Rapid-acting insulin analogues are now being used as alternatives to conventional insulins; their role in the management of type 2 diabetes mellitus is presently uncertain but reports of a reduced frequency of hypoglycaemia are encouraging. The development of new drugs aims to counter the principal metabolic defects of the disorder, respectively, relative insulin deficiency and insulin resistance. Novel classes of rapid-acting secretagogues under evaluation include the morphilinoguanide BTS 67582 and the meglitinides mitiglinide (KAD 1229) and senaglinide (A-4166). Succinate ester derivatives represent a potential novel approach to improving beta-cell function through enhancement of insulin biosynthesis and

  15. Treatment of Varicocele Infertility Men Patients of Different Chinese Medical Syndrome Types by Integrative Medicine Treatment Selection: a Primary Research%不同中医证型精索静脉曲张性不育症中西医治疗选择初探

    Institute of Scientific and Technical Information of China (English)

    倪凯; 陈斌; 李鹤; 王鸿祥; 杨昊; 胡凯; 韩银发; 王益鑫; 黄翼然

    2013-01-01

    treatment group (A) and the surgical group (B), each consisting of three Chinese medical syndrome types, i.e., damp-heat stagnation syndrome (DHSS), Shen-deficiency blood stasis syndrome (SDBSS), and blood stasis stagnation syndrome (BSSS), 20 in each group. Corresponding Chinese medical treatment was administered to those in Group A, C, and E, while microscopic ligation of internal vena spermatica was administered to those in Group B, D, and F. The routine analysis of semen, biochemical analysis of seminal plasma, and serum sex hormones (prolactin, testosterone, follicle stimulating hormone, luteinizing hormone, and estradiol) were performed before treatment and by the end of the 24th week after treatment. Results Totally 18 patients' spouses were pregnant. Of them, 1 in Group A of DHSS (abbreviated as Group A), 3 in Group B of DHSS (abbreviated as Group B), 4 in Group A of SDB- SS (abbreviated as Group C), 5 in Group A of SDBSS (abbreviated as Group D), 1 in Group A of BSSS (abbreviated as Group E), and 4 in Group B of BSSS (abbreviated as Group F). After 24-week treatment, the sperm concentration, class a sperm percentage, class a +b sperm percentage, the motility rate, the seminal plasma of fructose density, and the seminal plasma neutral alpha-glucosidase were more significantly improved in Group B, C, D and F, when compared with the same group before treatment (P 0. 05). As for the improvement percentage of seminal routine indices, the difference of the seminal plasma of fructose density, and the difference of seminal plasma neutral alpha-glucosidase between before and after treatment in the same Chinese medical syndrome types, better effects were obtained in Group B than in Group A (P 0. 05). There was no statistical difference in the 5 items of sex hormones in each group between before and after treatment (P >0. 05). Conclusions Surgical treatment could effectively improve the semen quality for male infertility VC patients accompanied with oligozoospermia or

  16. Analysis of hypoglycemic drugs of patients with type 2 diabetes in Qingxin Distric in Qingyuan city%清远市清新区2型糖尿病患者降糖药物的用药分析

    Institute of Scientific and Technical Information of China (English)

    罗金娥; 余德生; 姜学辉

    2016-01-01

    Objective:To provide reference for clinical rational drug use in the local, we investigate hypoglycemic drugs used in type 2 diabetic patients in Qingxin Distric in Qingyuan city in Guangdong Province. Methods:2380 cases of patients with type 2 diabetes in Qingxin Distric in Qingyuan city in Guangdong Province as the research object, questionnaire survey, and analyzed the use of antidiabetic drugs. Results:A higher proportion of drug use is: biguanides, using the rate of 89.97%. Secondly, a sulfonylurea, alpha glucosidase inhibitor, use rates were 71.97% and 55.97%. High frequency of use of drugs, metformin, acarbose, frequency of use are:79.03%, 57.02%. Double guanidine kind+sulfonylurea combination rate:71.47%, is the proportion of previous use of drug combination in the largest category. Followed by : biguanides + sulfonylurea + alpha glucosidase inhibitor drugs combination accounted for 50.63%. Fasting blood glucose (FBG) and lower levels of 2hPBG, were of relatively short duration, hypoglycemic drug varieties less. While the FBG and 2hPBG levels were higher in the patients, the disease is relatively longer, taking more hypoglycemic drugs. A short course of patients, complications and hypoglycemic drug relatively few species. While patients with longer disease duration, complications and use of hypoglycemic drugs more varieties. Conclusion:Hypoglycemic medication is more scientific and reasonable, accord with the clinical treatment of specification, type 2 diabetic patients in Qingxin Distric in Qingyuan city in Guangdong Province. Part of the control blood sugar of patients is not ideal by hypoglycemic drugs, can be used insulin treatment.%目的:调查分析广东省清远市清新区2型糖尿病患者的降糖药物使用情况,为临床合理用药提供参考。方法:选择广东省清远市清新区2380例2型糖尿病患者作为研究对象,发放调查问卷进行调查,并对降糖药物使用情况进行总结分析。结果:药物使

  17. 东北红豆杉枝叶不同提取部位体外降血糖活性研究%Study on the Hypoglycemic Activity of the Leaves and Twigs of Taxus Cuspidata Sieb et Zucc in Vitro

    Institute of Scientific and Technical Information of China (English)

    薛平; 姚鑫

    2016-01-01

    目的:探讨东北红豆杉枝叶不同提取部位对胰岛素抵抗HepG2细胞葡萄糖消耗量和抑制α-葡萄糖苷活性的影响。方法采用胰岛素抵抗HepG2细胞和α-葡萄糖苷酶作为体外受体模型,研究东北红豆杉枝叶不同提取部位的降血糖效果。结果对于胰岛素抵抗HepG2细胞,醇提液、乙酸乙酯部位和正丁醇部位均能促进胰岛素抵抗HepG2细胞的葡萄糖消耗量,其中醇提液和乙酸乙酯部位在浓度为0.01mg・ mL -1效果最佳,正丁醇部位在浓度为0.05mg・ mL -1效果最佳,而且都与模型组相比具有极显著性差异(P<0.01);东北红豆杉提取物均有不同程度抑制α-葡萄糖苷酶活性,以乙酸乙酯部位效果最佳(IC50=17.08mg・ L -1),其次正丁醇部位(IC50=28.48mg・ L-1)和醇提物(IC50=31.03mg・L -1),水部位(IC50=80.99mg・ L -1)石油醚部位(IC50=132.38mg・ L -1)最弱。结论初步确定东北红豆杉具有降血糖潜力的部位在乙酸乙酯与正丁醇部位。%ABSTRACT:OBJECTIVE To study the action of different extracts of Taxus cuspidata Sieb et Zucc on alpha-glu-cosidase and HepG2 cells′insulin resistance in vitro to supply the separating base of the effective chemical compo -nents.METHODS The HepG2 cells and alpha-glucosidase were used as receptor model to study the hypoglycemic activity in vitro of Taxus cuspidata Sieb et Zucc.RESULTS The ethyl acetate extrac(0.01mg・ mL-1)、ethanol ex-tract(0.01mg・ mL-1) and n-butanol extract(0.05mg・ mL-1) of Taxus cuspidata Sieb et Zucc could promote glu-cose consumption of insulin resistant HepG 2 cell model.The ethyl acetate extract showed strong activity in the inhibi-tory activities ofα-glucoside.( IC50 =17.08 mg・ L-1 ) The ethanol extract ( IC50 =31.03 mg・ L-1 ) and n-butanol ex-tract(IC50 =28.48mg・ L-1) displayed lower activity.Petroleum ether fraction(IC50 =132.38mg・ L-1) and

  18. Glucose-lowering treatment of type 2 diabetes. Part II--Glucose-lowering drugs after metformin: a choice based largely on adverse effects.

    Science.gov (United States)

    2015-05-01

    Metformin alone is the glucose-lowering drug of first choice for patients with type 2 diabetes. None of the other glucose-lowering drugs available in 2014 have any proven efficacy in preventing diabetes complications. How important are adverse effects in the choice of glucose-lowering alternatives to metformin for patients with type 2 diabetes? What about their effects on HbA1c levels? To answer these questions, we conducted a review of the literature using the standard Prescrire methodology. Sulphonylureas have been in use for many years. These drugs lower HbA1c levels by an average of 1.5% when used alone, and by 0.8% to 1% when added to metformin. All sulphonylureas can cause dose-related hypoglycaemia. Available data do not rule out a tangible increase or decrease in cardiovascular mortality among patients treated with sulphonylureas. Comparative data suggest that the combination of metformin + sulphonylurea increases overall mortality. Human insulins have also been in use for many years. A daily injection of long-acting insulin, added to on-going oral glucose-lowering therapy, lowers HbA1c by 0.7% to 2.5% on average but causes weight gain and increases the risk of hypoglycaemia. It cannot be ruled out that insulin may increase the risk of certain cancers. Alpha-glucosidase inhibitors have a weak glucose-lowering effect. The average decline in HbA1c is about 0.7%, which is not sufficient to offset the gastrointestinal disorders caused by these drugs. The glucose-lowering effect of repaglinide is similar to that of sulphonylureas. Repaglinide can cause hypoglycaemia, particularly when co-administered with inhibitors of some cytochrome P450 isoenzymes. Glitazones have a clearly unfavourable harm-benefit balance, potentially causing fractures, heart failure, other cardiovascular events, bladder cancer. Gliptins lower HbA1c by 0.7% on average but can provoke anaphylactic reactions, Stevens-Johnson syndrome, and infections. Saxagliptin may increase the risk of

  19. Antihyperglycemic, antihyperlipidemic, anti-inflammatory and adenosine deaminase–lowering effects of garlic in patients with type 2 diabetes mellitus with obesity

    Directory of Open Access Journals (Sweden)

    Kumar R

    2013-01-01

    Full Text Available Rahat Kumar,1 Simran Chhatwal,1 Sahiba Arora,2 Sita Sharma,3 Jaswinder Singh,1 Narinder Singh,1 Vikram Bhandari,1 Ashok Khurana41Department of Pharmacology, 2Department of Biochemistry, 3Department of Obstetrics and Gynecology, 4Department of Medicine, Sri Guru Ram Das Institute of Medical Sciences and Research, Amritsar, IndiaIntroduction: Type 2 diabetes mellitus is a chronic disorder characterized by chronic hyperglycemia, with long term macrovascular and microvascular complications. The treatment is lifestyle management, exercise, weight control, and antihyperglycemic drugs such as sulfonylureas, biguanides, alpha-glucosidase inhibitors, thiazolidinediones, and meglitinide. Recently, a direct association between high levels of C-reactive protein and serum adenosine deaminase levels in patients with uncontrolled diabetes with long-term complications has been seen. This study was conducted to assess the antihyperglycemic, lipid-lowering, anti-inflammatory, and improving glycemic control of garlic in type 2 diabetes patients with obesity.Materials and methods: This was an open-label, prospective, comparative study, conducted on 60 patients having type 2 diabetes mellitus and obesity. The patients were divided into two groups of 30 each, of either sex. Group 1 was given metformin tablets, 500 mg twice a day (BD/three times a day (TDS, after meals, and group 2 was given metformin tablets, 500 mg BD/TDS, after meals, along with garlic (Allium sativum capsules, 250 mg BD. Patients were routinely investigated for fasting and postprandial blood glucose, glycosylated hemoglobin (HbA1c, serum adenosine deaminase levels and lipid profile (serum cholesterol, high-density lipoprotein cholesterol, triglycerides and low-density lipoprotein cholesterol at the start of the study. Patients were followed up for 12 weeks, with monitoring of fasting and postprandial blood glucose at 2 week intervals, and monitoring of the other parameters at the end of study

  20. Analysis of Application of Oral Hypoglycemic Drugs in Outpatient Department of the Central Hospital of Nanchong during 2012-2014%2012—2014年南充市中心医院门诊口服降糖药应用分析

    Institute of Scientific and Technical Information of China (English)

    徐秀华; 冯婧; 王龙飞

    2015-01-01

    目的:分析南充市中心医院(以下简称"我院")2012—2014年门诊口服降糖药的应用情况和发展趋势,为糖尿病药物治疗提供参考.方法:回顾性统计分析我院2012—2014年门诊降糖药的销售数量、销售总金额、用药频度( defined daily dose system,DDDs)、限定日费用( daily drug cost,DDC),并根据各药的DDDs排序和销售金额排序,算出排序比.结果:我院门诊口服降糖药销售金额整体呈上升趋势,磺酰脲类、双胍类、α-葡萄糖苷酶抑制剂的销售金额增幅大;二甲双胍使用频率高,连续3年居第1位;二甲双胍日平均费用较低,排序比连续3年大于1.结论:我院2012—2014年门诊口服降糖药临床使用基本合理,二甲双胍使用频率高,符合安全、有效、经济的用药原则.%OBJECTIVE:To analyze the status quo and development tendency of oral hypoglycemic drugs for outpatients in the Central Hospital of Nanchong( hereinafter referred to as "our hospital") during 2012-2014,and to provide reference for the medication of diabetes.METHODS: Retrospective analysis was conducted in terms of the consumption amount,consumption sum,defined daily dose system ( DDDs ),and daily drug cost ( DDC ); and the ranking ratio was calculated based on the ranking of DDDs and medication cost of oral hypoglycemic drugs.RESULTS:The consumption sum of oral hypoglycemic drugs for outpatients in our hospital showed an increase tendency in general,especially for sulfonylurea,biguanides and alpha glucosidase inhibitors.The use frequency of metformin continuously dominated the first place during the three years,which had a low cost per day,and the ranking ratio was over 1 during the three years.CONCLUSIONS: The application of oral hypoglycemic drugs for inpatients in our hospital is basically rational during 2012-2014,and metformin has a high frequency of usage,which is conform to the principle of safe,effective and economic medication.

  1. Study on Inhibition of Alpha-glucoside and Antioxidation of from Leaves and Twigs of Taxus chinensis var mairei in vitro%南方红豆杉枝叶不同提取部位抑制α-葡萄糖苷酶及抗氧化活性筛选

    Institute of Scientific and Technical Information of China (English)

    薛平; 汤彬; 陈建伟; 王玉; 姚晓; 李祥

    2013-01-01

    目的:考察南方红豆杉枝叶不同提取部位体外抑制α-葡萄糖苷活性和抗氧化活性.方法:对南方红豆杉的醇提液、石油醚部位、乙酸乙酯部位、正丁醇部位、水部位采用体外α-葡萄糖苷酶抑制模型进行酶抑制活性研究,并通过其清除1,1二苯基苦基苯肼(DPPH)自由基和Fe3±还原能力(FRAP)测定南方红豆杉提取物的体外抗氧化作用.结果:南方红豆杉提取物均有不同程度抑制α-葡萄糖苷酶活性,以乙酸乙酯部位效果最佳(IC50 27.854 1 mg·L-1),其次醇提物(IC5029.215 5 mg·L-1)和正丁醇部位(IC50 38.913 0 mg·L-1),水部位(IC50 74.505 9 mg·L-1)石油醚部位(IC50 128.729 5 mg·L-1)最弱.抗氧化活性依次为醇提液(DPPH IC506.370 7 mg·L-1,FRAP 4.266 mmol·g-1)>乙酸乙酯部位(DPPH IC50 9.535 8 mg·L-1,FRAP 3.275 mmol·g-1)>正丁醇部位(DPPH IC50 20.461 3 mg·L-1,FRAP 1.498 mmol·g-1)>水部位(DPPH IC5026.685 5 mg·L-1,FRAP 0.678 mmol·g-1)>石油醚部位.结论:初步确定南方红豆杉抑制α-葡萄糖苷酶活性有效成分主要在乙酸乙酯与正丁醇部位,抗氧化及其相关成分主要在乙酸乙酯部位.%Objective: Inhibition of alpha-glucosidase and antioxidation of different extracts from the branches and leaves of Taxus chinensis var mairei were studied in vitro to supply the separating base of the effective chemical components. Method: The inhibitory activities of α-glucoside were assayed in external. The antioxidant activities of these extracts were assessed bytwo complementary test systems, namely 1, l-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and ferric reducing antioxidant power (FRAP) assay. Result: The ethyl acetate extract showed strong activity in the inhibitory activities of a-glucoside. ( IC50 27. 854 1 mg ·L-1 ) The ethanol extract ( IC50 29. 215 5 mg·L-1) and n-butanol extract ( IC50 38. 913 0 mg·L-1) displayed lower activity. Petroleum ether fraction (IC50 128.729 5 mg

  2. 亚洲来得时治疗达标研究(ATLAS)在中国2型糖尿病管理中的潜在价值:研究原理及设计%Potential value of the Asian Treat to Target Lantus Study (ATLAS) for the type 2 diabetes management in China: the rationale and design of ATLAS

    Institute of Scientific and Technical Information of China (English)

    潘长玉代表ATLAS中国研究组

    2011-01-01

    Asia and Russia patients with type 2 diabetes mellitus.Total 554 subjects are planned for this study and 160 subjects will be recruited in China.As one of the most important participating countries,China has some special characteristics in diabetes management,such as less self monitoring of blood glucose (SMBG),fewer self-titration and preferred premixed insulin in stead of basal insulin,et al.This report is aimed to introduce the study rationale and design of ATLAS with the situation in China.Methods Subjects (40-75 years old,body mass index ≥20 kg/m2 and ≤40 kg/m2 ) with type 2 diabetes mellitus for> 2 years,suboptimally controlled ( HbA1c ≥ 7.0% and ≤ 11.0% ) with stable doses of 2 oral anti-diabetic drugs (OADs) (sulphonylureas,biguanides,alpha-glucosidase inhibitors,DPP-IV inhibitors,and metiglinides)for>3 months,and not using insulin.The subjects will be randomized either to the patient-led or the physician-led titration arm for6 months treatment of glargine plus OADs regimen,where the insulin dose will be adjusted to achieve a target fasting plasma glucose value of 110 mg/dl (6.1 mmol/L).Study endpoints:The primary endpoint will be change in mean HbA1c at 6 months from baseline.For the primary endpoint,if non-inferiority is achieved using a HbA1c 0.3% boundary,superiority test will be undertaken.Secondary endpoints will include:proportion of patients reaching the target of HbA1c <7% with or without hypoglycemia; the number of patients whose HbA1c decreases at least 10% and/or 5% ; the change of fasting plasma glucose,postprandial plasma glucose,weight,dose of insulin glargine,patient treatment satisfaction,and quality of life in the two groups.Conclusions The ATLAS trial will provide information on the relative safety and efficacy of a patient-led versus physician led titration strategy for insulin glargine-based basal insulin initiation in patients who are not controlled with two OADs.The results of the study may provide new ideas and

  3. 婴儿型糖原累积症Ⅱ型六例临床分析%Analysis of clinical features of 6 patients with infantile type glycogen storage disease type Ⅱ

    Institute of Scientific and Technical Information of China (English)

    丁娟; 黄昱; 杨海坡; 张清友; 侯新琳; 刘雪芹; 杨艳玲; 能晖

    2015-01-01

    Objective To summarize clinical features and diagnosis of Chinese infantile patients with glycogen storage disease type Ⅱ (GSD Ⅱ).Method Six infant patients with GSD Ⅱ diagnosed from January 2012 to June 2014 in the Department of Pediatrics,Peking University First Hospital were enrolled into this study.Clinical information of the 6 patients,including clinical manifestation,blood biochemistry,chest X-ray,echocardiogram,electrocardiogram,acid alpha-glucosidase (GAA) activity and GAA gene mutation analysis by direct sequencing of polymerase chain reaction (PCR) product were reviewed.Result Of the 6 patients,five were female and one was male,five of whom were classic infantile type while the other one was atypical.The age of onset ranged from birth to 3-month-old.All patients had varying degrees of generalized muscle weakness,hypotonia and development retardation or retrogression.Other common findings were feeding difficulties in two patients,tongue weakness in two patients,respiratory distress in four patients,macroglossia in one patient,and hepatomegaly in two patients.Left ventricular hypertrophy and cardiomegaly were obvious in all the six patients.All six patients were found to have a enlarged heart in physical examination,and three patients who underwent a chest X-ray examination had an enlarged heart shadow.Four patients who had an echocardiography were found to have myocardial hypertrophy.The electrocardiogram in three patients showed short PR intervals and high voltage.The creatine kinase (CK) levels were three to seven times elevated.The mildest elevated CK was 441 IU/L,and the highest CK level was 1 238 U/L.Assay of GAA enzyme activity in whole blood showed significantly reduced activity (1.3 nmol/(spot · d) to 2 nmol/(spot · d)) in the patients tested.Gene sequencing in 4 patients showed 8 pathogenic mutations,including 6 missense mutations,one nonsense mutation and one frameshift mutation.The missense mutations were c.998C > A(p.Thr333Lys),c.1280