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PubMed1987

Immunological and radioimmunological studies in food allergy.

Experiments in order to induce food allergy were carried out in guinea pigs. The sensitization with egg albumin, pasteurized cow milk and bovine serum albumin provoked anaphylactic shock. The passive cutaneous anaphylaxis, serum antibodies, liver cytochrome P-450 concentration and the anaphylactic shock were determined. Some correlation between the mortality, anaphylactic antibodies and cytochrome P-450 monooxygenase system was established. The morphology of the jejunal mucosa, the activities of the 5 disaccharidases, the number of immunoglobulin secreting cells (Ig SC) and the mastocytes were investigated in 35 patients with food allergy. Normal mucosa was found in 28 cases as well as a significant decrease of the lactase, sucrase and trehalase activities. An increase of IgM and IgG secreting cells and of mastocytes, different electron microscopic changes in the enterocytes (an increased number of lysosomes, appearance of vesicles in cytoplasma, shortening, enlargement and uneven distribution of microvilli) as well as symptoms of functional activity in the plasmocytes and some others were also revealed. The experimental model obtained is similar to that one in humans according to the enteral way of sensitization the high selectivity of the allergic reaction which is of reagin type as the immunoglobulin changes are involved.

Animals↗
PubMed1986

[Biochemical manifestations of the poisoning of larvae of Aedes aegypti (Insecta, Diptera) by the delta-endotoxin of Bacillus thuringiensis israelensis. I. Hemolymph carbohydrates].

The glycemia of 4th instar larvae of Aedes aegypti (Diptera Culicidae) was determined over a 30 min to 36 hrs period following their immersion in water containing 0.00-0.01-0.02-0.1-1.0 mg per liter of a 12,000 I. U. per mg preparation of the delta-endotoxin of Bacillus thuringiensis israelensis. Seven major carbohydrate fractions were separated in the haemolymph, among which glycogen, trehalose and glucose were identified; the most concentrated of the 4 remaining unknown fractions, exhibits a similar chromatographic behavior but different chemical properties than glucuronic acid. The glycogen fraction shows two large amplitude peaks at 1 hr 30 and 6 hrs with 0.1 mg/l, whereas with the 2 lower doses (0.01 and 0.02 mg/l) a first drop from 30 min to 1 hr is followed by a peaking at 12 hrs. The haemolymph trehalose concentrations of intoxicated larvae increase relatively to controls 2 hrs after incubation with the lower dose; by contrast, they decrease with the 2 higher doses. The glucosemia increases in all cases, but more markedly with the higher dose as soon as 1 hr following the treatment. A positive correlation was found over the whole treatment duration between the glucosemia nd the trehalosemia of control larvae. The regression slope of this relationship decreases with increasing toxin doses and turns to negative values with 0.1 and 1.0 mg/l. These observations suggest the hypothesis that a uncontrolled increase of trehalase activity develops according to the increasing stages of intoxication. This clearly differenciates the effects of the B. T. i toxin from those of parasites, such as Fungi, Sporozoans or Nematodes in Mosquitoes.

Aedes↗
PubMed1987

Inhibition of intestinal disaccharidases and suppression of blood glucose by a new alpha-glucohydrolase inhibitor--MDL 25,637.

MDL 25,637 is a novel compound designed as a transition-state inhibitor of alpha-glucohydrolases. This compound inhibits rat intestinal sucrase, maltase, isomaltase, glucoamylase and trehalase activities at micromolar concentrations. It is a much weaker inhibitor of alpha-amylase and lactase. Inhibition of sucrase was competitive with sucrose. In mice, MDL 25,637 inhibited the rise in serum glucose after a sucrose or starch load but not after a glucose load. MDL 25,637 also reduced the glycemic response to sucrose in rats. The drug was most effective when administered 0 to 30 min before the sucrose load and was as effective in streptozotocin-treated rats as in normals. The inhibition by MDL 25,637 of intestinal glucohydrolases is an effective means of reducing the hyperglycemic response to an oral sucrose or starch load and, as such, warrants further investigation as a potential drug for the treatment of diabetes mellitus.

Animals↗
PubMed1985

Effect of growth and sodium butyrate on brush border membrane-associated hydrolases in human colorectal cancer cell lines.

The activities of brush border membrane-associated hydrolases such as alkaline phosphatase (Alkpase), aminopeptidase, dipeptidyl aminopeptidase IV (DAP-IV), sucrase, lactase, and trehalase were studied in 14 different human colorectal cancer cell lines. The effect of sodium butyrate, a known differentiating agent, and cell growth on the activities of these enzymes was also examined. All 14 cell lines exhibited brush border membrane enzyme activities, and in general, the activity of Alkpase, aminopeptidase, and DAP-IV was much higher than the disaccharidases. However, the specific enzyme activities varied among different cell lines. The induction of Alkpase activity by sodium butyrate occurred in most of the 14 cell lines (2- to 123-fold), while induction of the other enzyme activities was observed in several (1.5- to 3.5-fold). In some instances, butyrate caused a decrease in enzyme activity. There was no statistically significant correlation between the induction of Alkpase activity and that of other enzyme activities by sodium butyrate. Levels of aminopeptidase and DAP-IV activity were found to be dependent on cell density and increased 3- to 4-fold by the tenth day in most of the cell lines. Sodium butyrate altered the subcellular distribution pattern of the disaccharidases, causing a significant increase in activity associated with the soluble (cytoplasmic) fraction. Other enzymes such as Alkpase and DAP-IV continued to be predominantly associated with the membrane fraction in butyrate-treated cells. These data suggest that brush border membrane hydrolase activity and the effect of sodium butyrate may provide useful information regarding the differentiation of human colorectal cancer cells.

Alkaline Phosphatase↗
PubMed1979

Preparation of urine for enzyme determinations by gel filtration.

The activities of several enzymes in urine are masked by the presence of interfering substances in native urine. From several methods proposed for the removal of low molecular mass interferences dilution, dialysis, gel filtration, and ultrafiltration have been successfully applied. Gel filtration seems to be of these most suitable. I is effective, accurate, precise and economical. Scale-down procedures provide for acceptable speed. By this method the complete separation of lactate dehydrogenase, gamma-glutamyltransferase, alkaline phosphatase, arylsulphatase A, alpha-glucosidase, beta-galactosidase, trehalase, N-acetyl-beta-glucosaminidase, beta-glucuronidase and leucine arylamidase from low molecular mass substances, e.g. a heat-stable, competitive inhibitor of N-acetyl-beta-glucosaminidase was possible. The preparation and determination of urinary enzymes should be thoroughly standardized and controlled. Acceptable precision (coefficient of variation less than 10% between-day) can be achieved with manual spectrophotometric methods.

Chromatography, Gel↗
PubMed1979

Regional variability in circadian rhythmicity of intestinal digestive-absorptive functions.

The circadian rhythms of sucrase, maltase, isomaltase, trehalase, lactase, gamma-glutamyltransferase, leucylnaphthylamide hydrolyzing activity, alkaline phosphatase and monosaccharide transport were assessed in each fifth of the small intestine of the rat in order to determine if an entire enzyme or transport system population responded in a similar manner or if there were regional differences. Animals were maintained under a light-dark cycle and fed from 1400-1800, EST for 7 days. Functional activities were assessed every 4 h for 24 h, inclusively. Quantitative, and in a few instances, qualitative differences in different areas of the intestine were found for all functions. There were portions of the lactase and alkaline phosphatase populations which displayed no rhythmicity in activity. When rhythmicity was observed there were differences in the activity patterns along the intestine for all functions. Thus, the rhythm patterns obtained from homogenates of the entire small intestine are a composite of the patterns in regions of high average activity. Also, there appears to be a reasonable amount of local control of the various functions.

Alkaline Phosphatase↗
PubMed1984

Characterization of cyclic AMP-requiring yeast mutants altered in the catalytic subunit of protein kinase.

The cyr2 mutant of yeast, Saccharomyces cerevisiae, required cAMP for growth at 35 degrees C. The cyr2 mutation was suppressed by the bcy1 mutation which resulted in deficiency of the regulatory subunit of cAMP-dependent protein kinase. The DEAE-Sephacel elution profile of cyr2 cAMP-dependent protein kinase was markedly different from that observed for the wild-type enzyme. With histone as substrate, the cAMP-dependent protein kinase activity of cyr2 cells showed 100-fold greater Ka value for activation by cAMP at 35 degrees C than that of the wild-type cells, while the Kd value for cAMP of the mutant enzyme was not altered. The electrophoretic character, molecular weight, and pI value of the regulatory subunit of the mutant enzyme were the same as those of the wild-type enzyme. When histone, trehalase, and glutamate dehydrogenase were used as substrate, the free catalytic subunit of the mutant enzyme showed a markedly decreased affinity for ATP and was more thermolabile compared to that of the wild-type enzyme. The results indicated that the cyr2 phenotype was produced by a structural mutation in the cyr2 gene coding for the catalytic subunit of cAMP-dependent protein kinase in yeast.

Carrier Proteins↗
PubMed1977

Possibilities for the cytochemical diagnosis of enzymopathies.

The authors review the contemporary uses of histochemistry for the diagnosis of enzymopathies. Enterokinase, lactase, sucrase and trehalase deficiency can be diagnosed by histochemical methods. In glycogenoses, glycogen storage and glucose-6-phosphatase, acid alpha-glucosidase and phosphorylase deficiencies can be demonstrated. In mucopolysaccharidoses, the accumulation of acid muco-substances and changes in lysosomal enzyme activities can be demonstrated.

Biopsy↗
PubMed1984

Morphological identification of simulium sanctipauli and S. yahense in Liberia and comparison of results with those of enzyme electrophoresis.

Simulium sanctipauli and S. yahense have been incriminated as the main vectors of Onchocerca volvulus in the rain-forest zone of Liberia. An accurate identification of the females of the two species which breed in different types of water-courses but are morphologically similar is essential for a proper study of their epidemiological importance and the planning of control measures. Various external characters were examined for their diagnostic value by the comparison of identifications using these characters and those based on enzyme electrophoresis (phosphoglucomutase, trehalase). It was shown that the females of the two species can be separated reliably by the use of external morphological characters. The occasional finding in the same fly of phosphoglucomutase variants typical for each species indicated the possible occurrence of natural hybridization between S. sanctipauli and S. yahense.

Animals↗
PubMedDate not supplied

[Comparative study of mycobactin-dependent strains of mycobacteria isolated from the wood-pigeon with Mycobacterium avium and M. paratuberculosis: study of biological and antigenic characteristics].

In this investigation three mycobacterial strains isolated in our laboratory from wood-pigeons were compared with one strain isolated by Matthews and another by Jorgensen from, respectively, a wood-pigeon and a roe-deer. The strains were also compared with various strains of Mycobacterium avium and M. paratuberculosis. The strains isolated from the wood-pigeons formed a relatively homogeneous group, which could be distinguished from M. avium and M. paratuberculosis. It was interesting to verify that most of the cultural and biochemical properties of the wood-pigeon mycobacteria were similar to those of M. paratuberculosis. The strains formed rough colonies and grew slowly in special mediums containing M. phlei extracts or mycobactin. However, one must recall that mycobactin dependence was also reported by Matthews for some strains of M. avium. The tween hydrolysis test (10 days), negative for M. avium, was positive for both the wood-pigeon mycobacteria and M. paratuberculosis. The trehalase test, which appears to be regularly positive for M. avium, was also positive for the wood-pigeon mycobacteria and M. paratuberculosis. In respect to drug susceptibility, no significant differences were observed. The organisms were resistant to most drugs studied, but were also resistant to cycloserine, in contrast to most M. avium strains. On the other hand, the organisms were not distinguished using sensitins, prepared from wood-pigeon mycobacteria and M. paratuberculosis using specifically sensitized guinea-pigs. However, the wood-pigeon mycobacteria could be clearly differentiated from M. paratuberculosis and M. avium using serology methods. Indeed, the wood-pigeon mycobacteria did not agglutinate in the presence of any of the sera defining the M. avium-intracellulare serovars (serovars 1 to 28); and by immunodiffusion in agar the wood-pigeon mycobacteria antigens formed two precipitation lines which were absent from M. paratuberculosis. Judging from our findings, the wood-pigeon mycobacteria are more closely related to M. paratuberculosis than to M. avium. It appears that biochemical and antigenic properties are not sufficient to completely differentiate these bacteria. Further studies are needed, and we plan to investigate in the near future their pathogenicity for rabbits, chicken and calves.

Animals↗
PubMed1978

Experimentally induced intestinal metaplasia in Wistar rats by X-ray irradiation.

The gastric region of 5-week-old female Wistar rats was irradiated daily with 500 rad of X-ray up to a total of six times. Goblet cells and marker enzymes of the small intestine, such as lactase, trehalase, and maltase, appeared in the pyloric region of the glandular stomach of the rats from the 1st week after final irradiation. Intestinal type crypt without Paneth cells was observed from the 8th week. Sucrase activity appeared from the 26th week. Intestinal metaplasia with Paneth cells appeared from the 71st week. The number of goblet cells, intestinal type crypts, and Paneth cells increased with age. Gastric adenocarcinoma did not develop after irradiation.

Animals↗
PubMed1981

Control of disaccharidase activities in brush-border membranes of guinea pig fetuses: a role of pancreatic proteases?

The intestinal brush-border disaccharidases most resistant to pancreatic protease digestion in vitro are lactase and trehalase. When compared to maltase and sucrase, they are also those which showed the largest increase during development of guinea pig fetuses. These results suggest that pancreatic proteases may play a role in the control of brush-border disaccharidase activities during fetal development.

Animals↗
PubMed1993

[Early diagnosis of gastric cancer].

Endoscopic diagnosis completed by biopsy achieved remarkable accuracy. New trends--endoscopy with the use of ultrasound orange red porphyrin fluorescence elicited by blue light e. g. krypton laser and strip biopsy not only greater accuracy of endoscopic methods but also open up new therapeutic possibilities. Histochemical and histoenzymatic methods allow to classify histological findings of the norm (presence of pepsinogen and neutral glycoprotein) of atrophic gastritis with intestinal metaplasia (acid glycoprotein, activity of sucrose, alkaline phosphatase-AP, leucin-aminopeptidase-LAP) and of dysplasia (presence of sulfomucin, decrease of AF, LAP, sucrose and trehalase activity) and to divide them into well defined groups. We are able to distinguish which mucosal changes are suspect, we know that malignant transformation can after a certain period of time be expected approximately 2-14% yet we still do not know whether the period from dysplastic change to malignant transformation is not longer than malignant change in resected stomach. Epidemiologic and experimental data gives us enough reasons for dietary and chemopreventive measures (apart of other treatment) especially in risk groups patients.

Humans↗
PubMed2000

Carbohydrate metabolism during starvation in the silkworm Bombyx mori.

The effect of starvation on carbohydrate metabolism in the last instar larvae of the silkworm Bombyx mori was examined. Trehalose concentration in the hemolymph increased slightly during the first 6 h of starvation and decreased thereafter, whereas glucose concentration decreased rapidly immediately after diet deprivation. Starvation-induced hypertrehalosemia was completely inhibited by neck ligation, suggesting that starvation stimulates the release of a hypertrehalosemic factor(s) from the head. The percentage of active glycogen phosphorylase in the fat body increased within 3 h of starvation and its glycogen content decreased gradually. These observations suggest that production of trehalose from glycogen is enhanced in starved larvae. However, hypertrehalosemia during starvation cannot be explained by the increased supply of trehalose into hemolymph alone, as similar changes in phosphorylase activity and glycogen content in the fat body were observed in neck-ligated larvae, in which hemolymph trehalose concentration did not increase but decreased gradually. When injected into larvae, trehalose disappeared from hemolymph at a rate about 40% lower in starved larvae than neck-ligated larvae. The hemolymph lipid concentration increased during starvation, suggesting that an increased supply of lipids to tissues suppresses the consumption of hemolymph trehalose and this is an important factor in hypertrehalosemia.

Animals↗
PubMed2000

The Kluyver effect for trehalose in Saccharomyces cerevisiae.

Saccharomyces cerevisiae cells are able to grow using trehalose as a sole source of carbon and energy. However, the biomass yield obtained with trehalose was higher, and the specific growth rate lower, than that obtained with glucose or maltose. The respiratory inhibitor antimycin A prevented cell growth on trehalose, and no ethanol or glycerol was formed during batch growth on this carbon source. Thus, S. cerevisiae exhibits the KLUYVER effect for trehalose: this disaccharide is assimilated and respired, but, in contrast to glucose or maltose, it cannot be fermented. The high-affinity trehalose-H+ symporter encoded by the AGT1 gene is required for growth on trehalose. Analysis of the differences in the metabolism of maltose and trehalose (both disaccharides of glucose transported by active transport systems) indicated that the absence of trehalose fermentation is a consequence of low sugar influx into the cells during growth on this carbon source.

Antifungal Agents↗
PubMed2006

Use of the glucosidase inhibitor 1,4-dideoxy-1,4-imino-D-arabinitol as a probe to investigate hypertrehalosemic hormone-mediated change in carbohydrate and lipid metabolism in the cockroach, Periplaneta americana.

The glucosidase inhibitor 1,4-dideoxy-1,4-imino-D-arabinitol (DIA) was used to investigate the action of hypertrehalosemic hormone (HTH) on carbohydrate, neutral lipid, and phospholipid metabolism in the hemolymph and fat body of the cockroach, Periplaneta americana. DIA blocked the hypertrehalosemic and hyperglycemic effects of HTH, as well as the decrease in hemolymph neutral lipid and phospholipid normally induced by HTH. DIA diminished the accumulation of neutral lipid in the fat body formed under the influence of HTH and partly blocked the decrease in fat body phospholipid evoked by HTH. The increased incorporation of (14)C-glucose into fat body triacylglycerol in the presence of HTH was decreased by more than two-thirds when DIA was coinjected with the hormone. The results suggest that glucose derived from hemolymph trehalose is an important contributor to the formation of the glycerol backbone of newly formed triacylglycerol in the fat body.

Animals↗
PubMed1990

Effect of the ingestion of an extract from Bauhinia monandra seeds on rats.

Bauhinia monandra is a leguminous plant that would be used in human nutrition, however it shows a high hemagglutinating activity against trypsin-treated erythrocytes of rabbit, 2 ml of a NaCl-solution extract of the seeds was given to a group of rats for 10 days. Intestinal disaccharidase activity was determined. Histopathological and histochemical examinations were also performed. Lectins from Bauhinia monandra seeds did not cause any visible intestinal damage even when they seem to be able to bind to the enterocyte brush border. The low disaccharidase activity in the membrane and enterocyte fractions, which represent the enzyme synthesis level, indicates that lectins could reduce the synthesis of these enzymes rather than inhibit the active sites of the enzymes. This effect could be in turn related with the binding action of lectins, because no effect of the autoclaved extract on the disaccharidase activity was observed.

Animals↗
PubMedDate not supplied

The control of trehalose biosynthesis in Saccharomyces cerevisiae: evidence for a catabolite inactivation and repression of trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase.

During diauxic growth of yeast in glucose-rich medium, the accumulation of trehalose started well after complete exhaustion of glucose from the medium. The accumulation of the disaccharide was concomitant with a resumption of cell growth on the ethanol accumulated in the medium, but not with a degradation of glycogen which occurred as soon as glucose had been consumed. In contrast, in a mutant deficient in phosphoenolpyruvate carboxykinase, the synthesis of trehalose coincided exactly with the degradation of glycogen. Upon inoculation of stationary phase wild-type cells into a glucose medium, the activities of trehalose-6-phosphate (Tre6P) synthase and Tre6P phosphatase dropped in parallel to reach only 15% of their initial values after 3 h, and only recovered their original values as cells re-entered stationary phase. In the presence of cycloheximide, the decrease in Tre6P synthase and Tre6P phosphatase activities was restricted to 50-60%, the remaining decrease being inhibited by the drug. Furthermore, the reappearance of the enzyme activities following transfer of cells to an acetate medium was blocked by cycloheximide. It was also shown that loss of activity of these two enzymes required a combination of metabolizable sugars together with a nitrogen source. Low activities of Tre6P synthase and Tre6P phosphatase were measured in mutants with increased adenylate cyclase activity (RAS2ala18val19 mutants). Moreover, derepression of these enzymes at the approach of stationary phase was prevented in a pde2 mutant when it was cultivated in the presence of exogenous cyclic nucleotide. The mechanism of this effect is not clear, but may involve a transcriptional regulation by cAMP of the genes encoding these proteins.

Culture Media↗