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PubMed2005

Both heat-shock and cold-shock influence trehalose metabolism in an entomopathogenic nematode.

Heat-shock response is highly conserved in animals and microorganisms, and it results in the synthesis of heat-shock proteins. In yeast, heat-shock response has also been reported to induce trehalose accumulation. We explored the relationship between heat- (35 C) or cold-shock (1 and 10 C) and trehalose metabolism in the entomopathogenic nematode, Heterorhabditis bacteriophora. Because both heat- and cold-shocks may precede desiccation stress in natural soil environments, we hypothesized that nematodes may accumulate a general desiccation protectant, trehalose, under both situations. Indeed, both heat- and cold-shocks influenced trehalose accumulation and activities of enzymes of trehalose metabolism in H. bacteriophora. Trehalose increased by 5- and 6-fold in heat- and cold-shocked infective juveniles, respectively, within 3 hr of exposure, compared with the nematodes maintained at 25 C (culture temperature). The activity of trehalose-6-phosphate synthase (T6PS), an enzyme involved in the synthesis of trehalose, also significantly increased in both heat- and cold-shocked nematodes during the first 3 hr of exposure. Generally, the trehalose levels and activities of T6PS declined to their original levels within 3 hr when nematodes were transferred back to 25 C. In both heat- and cold-shocked nematodes, trehalase activity decreased significantly within the first 3 hr and generally returned to the original levels within 3 hr when these nematodes were transferred back to 25 C. The results demonstrate that the trehalose concentrations in H. bacteriophora are influenced by both heat- and cold-shocks and are regulated by the action of 2 trehalose-metabolizing enzymes, T6PS and trehalase. The accumulated trehalose may enhance survival of nematodes under both cold and warm conditions, but it may also provide simultaneous protection against desiccation that may result from subsequent evaporation or freezing. This is the first report of the relationship between trehalose metabolism and heat-shock for the Nematoda.

Animals↗

PubMed1990

Stimulation of carbohydrate metabolising enzymes by synthetic hypertrehalosemic peptides in thoracic musculature of the American cockroach, Periplaneta americana.

The ability of the synthetic hypertrehalosemic peptides, HT-I and HT-II, to influence the activities of glycogen phosphorylase, trehalase and hexokinase via elevation of Ca++ and cAMP levels was examined in thoracic musculature of the American cockroach, Periplaneta americana. The peptides effect dose- and time-dependent activation of phosphorylase, trehalase and hexokinase activities that occur concomitantly with elevated levels of intracellular calcium. In addition, HT-I increases the accumulation of cyclic AMP in muscle cells.

Amino Acid Sequence↗

PubMed1992

Disaccharidase activity in the small intestine of susceptible and resistant mice after primary and challenge infections with Giardia muris.

The activities of four disaccharidases were examined in resistant (C57Bl/6) and susceptible (C3H/HeN) mice during the primary infection with Giardia muris and after challenge with either trophozoite extract or cysts. Significant decreases in lactase, sucrase, trehalase, and maltase activities in C57Bl/6 mice and lactase and sucrase activities in C3H/HeN mice in the anterior 25% of the small intestine were observed on day 10 after infection. The activities of maltase, sucrase, trehalase, and lactase in the jejunum of C3H/HeN mice were significantly reduced after challenge with trophozoite extract, when compared with the uninfected or infected, but not challenged animals. Decreases in enzyme activities of C3H/HeN mice were evident as early as 12 hours after challenge with the extract. The resistant C57Bl/6 mice showed little change in disaccharidase activity after challenge with trophozoite extract. On the other hand, challenge with cysts resulted in a few decreases in disaccharidase activities in both strains of mice: C57Bl/6 mice showed decreases in the duodenum, while disaccharidases of C3H/HeN mice had lower activity more posteriorly. Thus, challenge with parasite antigen results in a more severe disaccharidase deficiency in susceptible hosts when compared with resistant ones.

Animals↗

PubMed1991

All eight possible mono-beta-D-glucosides of validoxylamine A. II. Biological activities.

The biological activities of all eight possible mono-beta-D-glucosides of validoxylamine A against Rhizoctonia solani were studied. The attachment of the D-glucosyl residue to validoxylamine A generally diminished the inhibitory activity against trehalase. The introduction of the D-glucosyl residue at the C-3 position did not cause serious loss in activity, while substitution at the C-6' position caused complete loss in trehalase inhibitory activity. Of the eight beta-D-glucosides, 4-O-beta-D-glucopyranosylvalidoxylamine A (4-O-beta-Glc-VA), 3-O-beta-Glc-VA and 5'-O-beta-Glc-VA exhibited very strong activity against R. solani in the "dendroid-test method". The antagonistic activity of sugars (1 mM) against validoxylamine A and 4-O-beta-Glc-VA was examined using the "dendroid-test method". The inhibitory effect of validoxylamine A on hyphal extension was not antagonized by any sugars tested, whereas that of 4-O-beta-Glc-VA was antagonized by beta-1,3- and beta-1,4-glucooligosaccharides. Of 2-O-, 3-O-, 4-O- and 7-O-beta-Glc-VAs, 7-O-beta-Glc-VA exhibiting the lowest activity was not antagonized by any beta-glucooligosaccharides tested. The inhibitory effect of 3-O- and 4-O-beta-Glc-VAs was antagonized by most beta-glucooligosaccharides. The uptake of 4-O-beta-Glc-VA into the mycelia was inhibited by laminaribiose and cellobiose but not by maltose.

Antifungal Agents↗

PubMed2004

Influence of juvenile hormone analogue on food consumption and digestive enzyme activities in Spodoptera mauritia Boisd.

Final instar larvae of S. mauritia treated topically on day 0, 1, 2 and day 3 with a daily dose of 20 microg juvenile hormone analogue (JHA) showed an increase in most of the nutritional parameters such as approximate digestibility, efficiency of conversion of ingested food, consumption index and growth rate. Also, the activities of digestive enzymes amylase, invertase, trehalase and protease increased significantly in JHA treated larvae. The supernumerary larvae formed after JHA treatments showed an increase in the activities of digestive enzymes. Neck-ligated larvae treated with 10 microg JHA exhibited a significant increase in the activities of trehalase and protease. The results demonstrate that treatments of JHA increase the activities of digestive enzymes in the last instar larvae of S. mauritia.

Amylases↗

PubMed2005

Regulation of trehalose metabolism by Adox and AdoMet in Saccharomyces cerevisiae.

Effect of a potent methylation inhibitor oxidized adenosine (Adox), and a universal methyl group donor S-adenosyl-L-methionine (AdoMet) on trehalose metabolism was studied in two haploids of S. cerevisiae of mating types MATalpha, met3 (6460 -8D) and MATa, leu2, ura3, his4 (8534 -10A). Trehalose level decreased in presence of Adox in both strains. Both neutral trehalase (NT) and trehalose-6-phosphate (tre-6-p) synthase activities increased in presence of Adox in -8D strain. Decrease in trehalose level in -8D thus could not be explained in the light of increased tre-6-p synthase activity; however, it could be correlated with increased NT activity. In strain -10A, NT activity was reduced in presence of Adox while tre-6-p synthase activity increased. Enzyme activity profiles in -10A thus do not explain the reduced trehalose level on Adox treatment. Effect of AdoMet was not very prominent in either strain, though in -8D a small increase in trehalose level was seen on treatment. Intracellular AdoMet level of untreated cells of -10A was seen to be almost six times higher than that of -8D. Further, AdoMet treatment caused increase in its level compared to untreated cells, suggesting AdoMet uptake. No effect of either compound was seen on acid trehalase (AT) activity in any strain. The results suggest that there was a possible effect of demethylation on trehalose metabolism (particularly in the synthetic direction) in both strains, though effect of methylation was not very prominent, the reason for which is not very clear.

Adenosine↗

PubMed2001

[The enzymes of carbohydrates metabolism from Cysttidicola farionis (Cystidicolidae)].

The content of glycogen, glucose and trehalose was measured in larvae and adults of Cystidicola farionis, the parasite isolated from the swim bladder of Osmerus eperlanus from Vistula Lagoon. Activity of glycogen phosphorylase, alpha-amylase, glucoamylase, maltase, trehalase, and trehalose phosphorylase were measured. The highest activity was recorded for alpha-amylase 10.07 +/- 0.97 mu/mg and 7.47 +/- 0.24 mu/mg, next maltase 1.34 +/- 0.63 micromol/mg and 2.06 +/- 1.65 micronol/mg respectively for larvae and adults. The activity of glucoamylase was nearly the same for adults and larvae (about 0.20 micromol/mg). The trehalase activity was higher at adults (0.49 +/- 0.42 micromol/mg) than at larvae (0.18 +/- 0.12 micromol/mg). The activity of glycogen phosphorylase was much higher at larvae (3.58 +/- 1.49 micromol/mg) than at adults parasite (0.10 +/- 0.02 micromol/mg). The trehalose phosphorylase was present in both stages of parasite, but its activity was low. The content of glycogen and glucose was two-times higher in the adults' body than in larvae.

Animals↗

PubMed1980

Distribution of marker enzymes and mucin in intestinal metaplasia in human stomach and relation to complete and incomplete types of intestinal metaplasia to minute gastric carcinomas.

Intestinal metaplasia of the human stomach was classified into two types, complete and incomplete. The complete type was associated with the intestinal marker enzymes sucrose alpha-D-glucohydrolase, alpha, alpha-trehalase, aminopeptidase (microsomal) (APM), and alkaline phosphatase (ALP). Tissue of this type contained goblet cells and Paneth's cells but not high-iron diamine (HID)-positive mucin staining with HID-Alcian blue. The incomplete type of intestinal metaplasia was associated with sucrose alpha-D-glucohydrolase, APM, goblet cells, and HID-positive mucin but not with alpha, alpha-trehalase, ALP, or Paneth's cells. For the examination of the distribution of the complete and incomplete types in 84, 27, and 16 resected specimens of human stomach with gastric carcinoma, gastric ulcer, and duodenal ulcer, respectively, disaccharidases were located with Tes-Tape. Specimens with intestinal metaplasia were divided into three classes: complete type only (class I), incomplete type only (class II), and a mixture of areas of the complete and incomplete types (class III). Of the 84 specimens from patients with gastric carcinoma, intestinal metaplasia was found in 76 (01%), and the percentages of specimens of classes I, II, and III were 32, 22, and 46, respectively. In these specimens, the percent incidence of class I increased and that of class II decreased with age. Of the 27 specimens from patients with gastric ulcer, 16 (59%) shopwed intestinal metaplasia and 10 of the 16 (63%) specimens were of class II. Of the 16 specimens from patients with duodenal ulcer, only 3 (19%) specimens showed intestinal metaplasia and all of them were of class II. The relationships of the complete and incomplete types of intestinal metaplasia to gastric carcinoma wre studied in 26 foci of minute carcinoma of the stomach less than 5 mm in largest diameter. Nineteen of 20 (05%) foci of the intestinal type of minute carcinoma were surrounded by intestinal metaplasia and 16 foci (80%) were surrounded by the incomplete type of intestinal metaplasia.

Adult↗

PubMed1995

Disaccharidase activity in male and female C57BL/6 mice infected with Giardia muris.

To elucidate the contribution of host and parasite factors in induction of small-intestinal disaccharidase deficiency in giardiasis, we determined the activity of four enzymes in male and female C57BL/6 mice infected with Giardia muris. Both male and female mice exhibited significant disaccharidase deficiency as shown by decreases in the activities of lactase, sucrase, trehalase and maltase on day 10 after infection. However, by 20 days after infection the females had normal enzyme activities, whereas those in males remained significantly reduced. Prolonged disaccharidase deficiency in the males was related to the course of the primary infection where males had higher parasite loads in the small intestine than did females on day 20 after infection. By day 40 after the primary infection the enzyme activities had returned to normal levels and were similar in male and female mice. Secondary exposure of mice to either the infective cysts or a crude extract of the trophozoites caused disaccharidase deficiency. The females had lower activities of sucrase and trehalase as compared with males after the challenge. Thus, during the primary infection, disaccharidase deficiency was strongly associated with parasite number, whereas after challenge infections the more resistant females had lower enzyme activities in the small intestine than did males.

Analysis of Variance↗

PubMed1994

Possible involvement of a phosphatidylinositol-type signaling pathway in glucose-induced activation of plasma membrane H(+)-ATPase and cellular proton extrusion in the yeast Saccharomyces cerevisiae.

Addition of glucose to cells of the yeast Saccharomyces cerevisiae causes rapid activation of plasma membrane H(+)-ATPase and a stimulation of cellular H+ extrusion. We show that addition of diacylglycerol and other activators of protein kinase C to intact cells also activates the H(+)-ATPase and causes at the same time a stimulation of H+ extrusion from the cells. Both effects are reversed by addition of staurosporine, a protein kinase C inhibitor. Addition of staurosporine or calmidazolium, an inhibitor of Ca2+/calmodulin-dependent protein kinases, separately, causes a partial inhibition of glucose-induced H(+)-ATPase activation and stimulation of cellular H+ extrusion; together they cause a more potent inhibition. Addition of neomycin, which complexes with phosphatidylinositol 4,5-bisphosphate, or addition of compound 48/80, a phospholipase C inhibitor, also causes near complete inhibition. Diacylglycerol and other protein kinase C activators had no effect on the activity of the K(+)-uptake system and the activity of trehalase and glucose-induced activation of the K(+)-uptake system and trehalase was not inhibited by neomycin, supporting the specificity of the effects observed on the H(+)-ATPase. The results support a model in which glucose-induced activation of H(+)-ATPase is mediated by a phosphatidylinositol-type signaling pathway triggering phosphorylation of the enzyme both by protein kinase C and one or more Ca2+/calmodulin-dependent protein kinases.

Alkaloids↗

PubMed2000

Homonojirimycin analogues and their glucosides from Lobelia sessilifolia and Adenophora spp. (Campanulaceae).

2,6-Dideoxy-7-O-(beta-D-glucopyranosyl) 2,6-imino-D-glycero-L-gulo- heptitol (7-O-beta-D-glucopyranosyl-alpha-homonojirimycin, 1) was isolated from the 50% methanol extract of the whole plant of Lobelia sessilifolia (Campanulaceae), which was found to potently inhibit rice alpha-glucosidase. Adenophorae radix, roots of Adenophora spp. (Campanulaceae), yielded new homonojirimycin derivatives, adenophorine (2), 1-deoxyadenophorine (3), 5-deoxyadenophorine (4), 1-C-(5-amino-5-deoxy-beta-D-galactopyranosyl)butane (beta-1-C-butyl-deoxygalactonojirimycin, 5), and the 1-O-beta-D-glucosides of 2 (6) and 4 (7), in addition to the recently discovered alpha-1-C-ethylfagomine (8) and the known 1-deoxymannojirimycin (9) and 2R,5R-bis(hydroxymethyl)-3R,4R- dihydroxypyrrolidine (DMDP, 10). Compound 4 is a potent inhibitor of coffee bean alpha-galactosidase (IC50 = 6.4 microM) and a reasonably good inhibitor of bovine liver beta-galactosidase (IC50 = 34 microM). Compound 5 is a very specific and potent inhibitor of coffee bean alpha-galactosidase (IC50 = 0.71 microM). The glucosides 1 and 7 were potent inhibitors of various alpha-glucosidases, with IC50 values ranging from 1 to 0.1 microM. Furthermore, 1 potently inhibited porcine kidney trehalase (IC50 = 0.013 microM) but failed to inhibit alpha-galactosidase, whereas 7 was a potent inhibitor of alpha-galactosidase (IC50 = 1.7 microM) without trehalase inhibitory activity.

1-Deoxynojirimycin↗

PubMed1998

Partial desiccation induced by sub-zero temperatures as a component of the survival strategy of the Arctic collembolan Onychiurus arcticus (Tullberg).

The mechanism by which the freeze susceptible Arctic collembolan Onychiurus arcticus survives winter temperatures of -25 degrees C in the field is not fully understood but exposure to sub-zero temperatures (e.g. -2.5 degrees C) is known to induce dehydration and lower the supercooling point (SCP). In this study, changes in the water status and certain biochemical parameters (measured in individual Collembola) during a 3-week exposure to decreasing temperatures from 0 to -5.5 degrees C were studied. Osmotically active and inactive body water contents were measured by differential scanning calorimetry (DSC), water soluble carbohydrates by high performances liquid chromatography (HPLC) and glycogen by enzymatic assays. The activity of trehalase and trehalose 6-phosphate synthase were also measured. During the experiment, total water content decreased from 70 to 40% of fresh weight, mostly by the loss of osmotically active water with only a small reduction in the osmotically inactive component. The SCP decreased from -7 to -17 degrees C. Analysis of the results shows that if O. arcticus is exposed to -7 degrees C in the presence of ice, all osmotically active water would be lost due to the vapour pressure gradient between the animals supercooled body fluids and the ice. Under these conditions the estimated SCP would reach a minimum of c. -27 degrees C, but the Collembola may never freeze as all the osmotically active water has been lost, the animal becoming almost anhydrobiotic. Trehalose concentration increased from 0.9 to 94.7&mgr;g mg(-1)fw while glycogen reserves declined from 160 to 7.7 nmol glucose equivalents mg(-1) protein. Trehalase activity declined as the temperature was reduced, while trehalose 6-phosphate activity peaked at 0 degrees C. By adopting a strategy of near anhydrobiosis induced by sub-zero temperatures, O. arcticus, which was previously thought to be poorly adapted to survive severe winter temperatures, is able to colonise high Arctic habitats.

Journal Article↗

PubMed1968

Carbohydrase activities in the bovine digestive tract.

1. The carbohydrase activities of homogenates of mucosa from the abomasum, small intestine, caecum and colon, and of the pancreas of cattle were studied. 2. The disaccharidase activities were located mainly in the small intestine and showed a non-uniform pattern of distribution along the small intestine; trehalase activity was highest in the proximal part, lactase and cellobiase activities were highest in the proximal and middle parts and maltase activity was highest in the distal part. 3. The intestinal lactase and cellobiase activities were highest in the young calf and decreased with age, whereas the intestinal maltase and trehalase activities, which were very low compared with the lactase activity, did not change with age. 4. No intestinal sucrase or palatinase activity was detected in the calf or in the adult cow. 5. Homogenates of intestinal mucosa also exhibited amylase and dextranase activity. 6. Homogenates of the pancreas possessed a strong amylase activity and a weak maltase activity. The maltase activity did not change with age, whereas the amylase activity increased with age. 7. No marked differences were observed between the carbohydrase activities of calves fed solely on milk and those of calves given a concentrate-hay diet from 6 weeks of age.

Abomasum↗

PubMed1979

Proteins of the kidney microvillar membrane. Immunoelectrophoretic analysis of the membrane hydrolases: identification and resolution of the detergent- and proteinase-solubilized forms.

Antibodies raised in rabbits to detergent-solubilized pig kidney microvillar proteins have been used to investigate the membrane hydrolases by crossed immunoelectrophoresis. Eight enzymes were detected by specific staining methods: aminopeptidase M, dipeptidylpeptidase IV, neutral endopeptidase, aminopeptidase A, carboxypeptidase P, gamma-glutamyltransferase, trehalase and phosphodiesterase I. The mobility of all these enzymes, with the exception of trehalase and neutral endopeptidase, was increased by treatment of the detergent-solubilized preparation with papain. The difference between the detergent and proteinase forms of these enzymes is attributed to the removal of a small, non-antigenic peptide to which detergent is bound in significant quantities. This interpretation was further supported by experiments in which the microvillus fraction was labelled with an intramembrane photolabelling reagent, 1-azido-4-[125I]iodobenzene. After photolysis, the radioactivity in the membrane could be solubilized by detergent treatment but not by papain treatment. Radioautography after crossed charge-shift immunoelectrophoresis showed a good correlation between charge-shift (signifying the presence of detergent bound to a hydrophobic domain) and the presence of the label.

Animals↗

PubMed2003

The Gap1 general amino acid permease acts as an amino acid sensor for activation of protein kinase A targets in the yeast Saccharomyces cerevisiae.

Addition of a nitrogen source to yeast (Saccharomyces cerevisiae) cells starved for nitrogen on a glucose-containing medium triggers activation of protein kinase A (PKA) targets through a pathway that requires for sustained activation both a fermentable carbon source and a complete growth medium (fermentable growth medium induced or FGM pathway). Trehalase is activated, trehalose and glycogen content as well as heat resistance drop rapidly, STRE-controlled genes are repressed, and ribosomal protein genes are induced. We show that the rapid effect of amino acids on these targets specifically requires the general amino acid permease Gap1. In the gap1Delta strain, transport of high concentrations of l-citrulline occurs at a high rate but without activation of trehalase. Metabolism of the amino acids is not required. Point mutants in Gap1 with reduced or deficient transport also showed reduced or deficient signalling. However, two mutations, S391A and S397A, were identified with a differential effect on transport and signalling for l-glutamate and l-citrulline. Specific truncations of the C-terminus of Gap1 (e.g. last 14 or 26 amino acids) did not reduce transport activity but caused the same phenotype as in strains with constitutively high PKA activity also during growth with ammonium as sole nitrogen source. The overactive PKA phenotype was abolished by mutations in the Tpk1 or Tpk2 catalytic subunits. We conclude that Gap1 acts as an amino acid sensor for rapid activation of the FGM signalling pathway which controls the PKA targets, that transport through Gap1 is connected to signalling and that specific truncations of the C-terminus result in permanently activating Gap1 alleles.

Amino Acid Sequence↗

PubMed2001

Diet and the evolution of digestion and renal function in phyllostomid bats.

Bat species in the monophyletic family Phyllostomidae feed on blood, insects, small vertebrates, nectar, fruit and complex omnivorous mixtures. We used nitrogen stable isotope ratios to characterize bat diets and adopted a phylogenetically informed approach to investigate the physiological changes that accompany evolutionary diet changes in phyllostomids. We found that nitrogen stable isotopes separated plant-eating from animal-eating species. The blood of the latter was enriched in (15)N. A recent phylogenetic hypothesis suggests that with the possible exception of carnivory, which may have evolved twice, all diets evolved only once from insectivory. The shift from insectivory to nectarivory and frugivory was accompanied by increased intestinal sucrase and maltase activity, decreased trehalase activity, and reduced relative medullary thickness of kidneys. The shift from insectivory to sanguinivory and carnivory resulted in reduced trehalase activity. Vampire bats are the only known vertebrates that do not exhibit intestinal maltase activity. We argue that these physiological changes are adaptive responses to evolutionary diet shifts.

Journal Article↗

PubMed2006

Inhibitory effects of extractives from leaves of Morus alba on human and rat small intestinal disaccharidase activity.

The inhibitory effect on human and rat intestinal disaccharidase by the extractive from the leaves of Morus alba (ELM) containing 0.24 % 1-deoxynojirimycin equivalent and its inhibitory activities were investigated by the modified Dahlqvist method. In the presence of 1000-fold diluted ELM solution, the sucrase activity of four human samples was inhibited by 96 % and that of maltase and isomaltase by 95 and 99 %, respectively. The activities of trehalase and lactase were inhibited by 44 and 38 %, respectively. The human disaccharidase activities varied from sample to sample because the samples were obtained from different resected regions after surgery. However, the ratio of the inhibitory effect for sucrase, maltase, isomaltase, trehalase and lactase was very similar among the four samples, and also that of resembled rat intestinal disaccharides. The inhibitory constant of the 1-deoxynojirimycin equivalent for sucrase, maltase and isomaltase was 2.1 x 10(-4), 2.5 x 10(-4) and 4.5 x 10(-4) mm, respectively, and these inhibitory activities were shown, using rat brush border membrane vesicles, to be competitive. These results demonstrate that digestion is inhibited when an appropriate amount of ELM is orally ingested with sucrose or polysaccharide in man. When ELM was orally administered in a sucrose solution to fasted rats, the elevation in blood glucose was significantly suppressed, depending on the concentration of ELM given. These results suggest that ELM could be used as an ingredient in health foods and in foods that help to prevent diabetes.

Animals↗

PubMed1998

Responses of intestinal and renal alpha-glycosidases to alloxan and streptozotocin-induced diabetes: a comparative study.

Experimentally induced diabetes in the rat resulted in an increased level of alpha-glycosidases in the intestine but a depression in their levels in the kidney. Rat intestine exhibited a differential stimulation of maltase, sucrase and trehalase activities. The variations depended on the duration of diabetes and the beta-cytotoxic compounds used i.e. alloxan and streptozotocin. The maximum elevation in terms of total units and specific activity was observed on the 30th day in the following order: maltase>sucrase>trehalase. A significant observation emerging from this study is that the level of intestinal enzymes increases while that of the kidney enzymes declined during the period. Although intestinal and renal alpha-glycosidases are known to be structurally and biochemically similar, their opposing responses to diabetes indicates that they are under different regulatory mechanisms in these tissues.

Animals↗