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The selectin family of carbohydrate-binding proteins: structure and importance of carbohydrate ligands for cell adhesion.

Protein-carbohydrate interactions have been found to be important in many steps in lymphocyte recirculation and inflammatory responses. A family of carbohydrate-binding proteins or lectins, termed selectins, has been discovered and shown to be involved directly in these processes. The three known selectins, termed L-, E- and P-selectins, have domains homologous to other Ca(2+)-dependent (C-type) lectins. L-selectin is expressed constitutively on lymphocytes, E-selectin is expressed by activated endothelial cells, and P-selectin is expressed by activated platelets and endothelial cells. Here, we review the nature of the carbohydrate determinants in tissues recognized by these selectins. The expression of specific sialylated, fucosylated and sulfated carbohydrates in activated endothelium and high endothelial venules promotes interactions with L-selectin on leukocyte surfaces. In contrast, E- and P-selectins recognize specific carbohydrate determinants related to sialyl Le(x) antigen on neutrophil and monocyte surfaces. The discovery of the selectins has generated excitement among glycoconjugate researchers that other carbohydrate-binding proteins and their cognate ligands will be found to function in regulating many types of cellular interactions.

Amino Acid Sequence

Milk carbohydrates of marsupials. II. Quantitative and qualitative changes in milk carbohydrates during lactation in the tammar wallaby (Macropus eugenii).

Milk was collected at various stages of lactation from a group of tammar wallabies, M. eugenii, in which parturition had been synchronized. The milk carbohydrate was determined by a phenol-sulfuric acid method which had been modified to give equal colour yields for galactose and glucose. The mean carbohydrate content increased gradually during the first 6 months of lactation to a peak of 13 g hexose/100 ml of milk, but then fell rapidly to much lower values, over the following 2 months. Throughouth lactation, galactose was the predominant monosaccharide constituent of acid hydrolysates of the milk carbohydrate. Glucose, glucosamine, galactosamine and sialic acid were the only other monosaccharides present. Qualitative changes were investigated by gel filtration and thin-layer chromatography. During the first 6 months post partum the milk carbohydrate was composed of a variety of oligosaccharides including lactose, but from 8 months onwards it consisted mainly of free monosaccharides. Between 6 and 8 months an intermediate pattern was observed, i.e. a mixture of lower oligosaccharides and free monosaccharides. In two animals which suckled both a new-born pouch young and a young at foot, the mammary gland supplying the new-born secreted milk which was rich in oligosaccharides, whereas that supplying the young at foot produced milk in which the carbohydrates were mainly free monosaccharides, and which had a much lower carbohydrate content.

Age Factors

Comparative studies in obese subjects fed carbohydrate-restricted and high carbohydrate 1,000-calorie formula diets.

45 obese subjects were fed a high-carbohydrate, relatively low-fat, or a low-carbohydrate, relatively high-fat 1,000-calorie (4.14MJ) formula diet. The diet provided for an isoenergetic substitution of 170 g of carbohydrates for 75 g of fat. Weight reduction up to day 30 was significantly higher in the subjects on the carbohydrate-restricted diet. There were no significant differences between the water and electrolyte balances. The mean total weight reduction achieved on the high-carbohydrate diet was 9.8 +/- 4.5kg with a mean daily weight loss of 298 +/- 80g, while the corresponding values on the carbohydrate-restricted diet were 14 +/- 7.2 kg and 362 +/- 91 g/day, respectively.

Adult

Comparison of the carbohydrate of Sinbis virus glycoproteins with the carbohydrate of host glycoproteins.

The carbohydrate portions of the Sindbis virus glycoproteins were compared with the carbohydrate portions of cell surface glycoproteins from uninfected host cells. Comparisons of the size of glycopeptides were made using gel filtrations. Comparisons of sugar linkages were made by methylation analysis. The conclusion was that the Sindbis carbohydrate is similar to a portion of the host carbohydrate. Thus, the Sindbis carbohydrate structures appear to be structures normally made in the uninfected host cell, but which are added to the Sindbis glycoproteins in virus-infected cells.

Animals

A rat bioassay for measuring the comparative availability of carbohydrates and its application to legume foods, pure carbohydrates and polyols.

An assay was developed to evaluate the bioavailability of dietary carbohydrate by slope-ratio analysis of weight gain and plasma ketones of rats fed a carbohydrate-free diet supplemented with glucose as a standard and selected food items, pure carbohydrates and polyols. The diet was based on 35% food grade oleic acid, 12% casein protein supplemented with methionine (0.3%) and arginine (0.2%), 7.5% soybean oil, 37% cellulose, minerals and vitamins. Glucose, as the reference standard, or test materials were incorporated in the diet by replacing equi-energetic amounts of oleic acid and adjusting cellulose to equalize total weight. Optimal levels of protein (casein) and triglyceride (soybean oil) in the basal diet were defined as those levels above which additional protein or fat would serve only to provide endogenous glucose from their glucogenic amino acids and glycerol constituents. A standard dose-response curve was obtained by feeding diets containing 0%, 1% and 2% glucose. Similar dose response curves were obtained by feeding test materials. Under these experimental conditions, additional glucose or additional protein were growth stimulating and casein had approximately 50% of the value of an equal weight of glucose, which was consistent with its content of glucogenic amino acids. The specific carbohydrate value of a food was estimated in the assay by subtracting the calculated glucogenic value of its digestible protein from the total response. The apparent availability of the carbohydrates (i.e., nitrogen-free extract) in soybean meal, lima bean (Phaseolus lumatus), and chick pea (Cicer arietinum) were 35, 70, and 80%, respectively, as available as glucose. Galactose, a 1:1 mixture of galactose + glucose, fractose and starch were as available as glucose. Lactose, ribose, sorbitol, and xylitol were 50 to 65% as available as glucose while mannitol and inulin were not available to a significant extent. Possible metabolic bases for these differences are discussed.

Amino Acids

[Activity of carbohydrate-metabolizing enzymes and content of carbohydrate and nitrogen compounds in muscles of young cattle depending on breed and sex].

The activity of glycolysis and glyconeogenesis enzymes, content of carbohydrate and nitrogen compounds were studied in muscles of 15-month cattle of different sex of the black-piebald breed and its crosses with bulls of two meat breeds Hereford and Limousine. In the muscles of bulls the activity of phosphoglucomutase, phosphohexoisomerase, aldolase and fructose-1,6-diphosphatase is considerably higher (except of the activity of phosphoglucomutase and phosphohexoisomerase in the muscles of pure-bred black-piebald animals for which difference is not statistically reliable) and the content of glycogen, glucose, fructose, lactic acid, free and phosphorylated pentoses of nonadenylic compounds is essentially lower than in the muscle tissue of heifers of analogous breed groups. A higher activity of carbohydrate metabolism enzymes (especially aldolase and fructose-1,6-diphosphate) and a higher content of total pentoses, the adenylic system pentoses, ATP phosphorus in the muscles of the cross Limousine and Hereford bulls and Limousine heifers as compared to the pure-bred black-piebald animals of the corresponding sex coincide with a greater increase in the muscular tissue and more intensive synthesis of proteins in it. A considerably lower level of glycogen, glucose, fructose and a relatively high activity of carbohydrate metabolism enzymes in the muscles of cross young cattle show that disintegration of these carbohydrates is in excess of their synthesis, that is due to an increase in the energy demand connected with a more intensive synthesis of proteins in the muscular tissue. Therefore, in the authors opinion the performed kill of the cross Limousine and Hereford bulls as well as Limousine heifers, is somewhat untimely and unreasonable. At the same time the activity of all the studied enzymes in the muscles of the cross Hereford heifers, vice versa, is considerably lower as compared to the black-piebald heifers and coincides with a low gain in live weight and muscular tissue, a more rapid accumulation of glycogen and lipids and a more delayed--of proteins in the muscular tissue, that evidences for their early maturity. Therefore the further raising of the cross Hereford heifers in the farm for obtaining meat is economically less profitable. The data obtained give grounds to recommend determination of the activity of carbohydrate metabolism enzymes, especially of aldolase and fructose-1,6-diphosphate, as a test for checking the muscular tissue growth and for prognosing the meat productivity in the growing pure-bred and cross young cattle of different sex.

Adenosine Triphosphate

[Changes in carbohydrate tolerance and birth weight during carbohydrate-reduced diet in latent diabetic pregnancy (author's transl)].

The extent to which glucose tolerance can be improved by a carbohydrate-reduced diet in pregnant women with latent diabetes was investigated and, furthermore, whether this measure can reduce the incidence of high birth weight in the infants. 1. In a group of pregnant women who were thought to have some carbohydrate, metabolic disturbance on the basis of adipositas, glycosuria, hereditary taint or a history of infants with birth weights over 4000 g, the oral glucose tolerance test (GTT), showed a significant improvement under diet during pregnancy in comparison with a control group without diet. 2. Although an increase in the mean GTT after birth is to be expected--and was observed in the control group--, under diet the GTT values actually decreased post partum. 3. The extent of improvement in the GTT values depended on the degree of carbohydrate reduction in the diet. 4. In the years 1968 to 1976 all pregnant women with latent (gestational) diabetes showing high GTT values were submitted to a carbohydrate-reduced diet. This measure contributed to a reduction in the incidence of birth weights over 4000 g from 7.8% to 4.8%.

Birth Weight

Carbohydrate inhibitors of concanavalin A that inhibit binding of insulin-sepharose to fat cells and antagonize and mimic insulin's bioactivity. A possible role for membrane carbohydrate in insulin's action.

A consistent pattern of insulin-like properties is expressed by a variety of glycoside inhibitors of concanavalin A (Con A), and is suggestive of a common mechanism of action to explain these effects. Various exogenously added glycoside derivatives inhibit the binding of insulin-Sepharose beads to insulin receptors on isolated intact rat fat cells with a specificity resembling that for Con A-Sepharose binding to these cells. A more limited number of glycosides tested were also found to inhibit the binding of 125I-insulin, although some enhancement of binding that preceded the inhibition was observed for some of these saccharides. The glycosides also antagonize insulin-stimulated glucose utilization by the cells, but in some cases also mimic the hormone by stimulating glucose utilization. A few glycosides mimic insulin without appearing to antagonize its bioactivity. Radiolabeled glycoside inhibitors fail to bind to insulin in equilibrium dialysis experiments although they readily bind to Con A, indicating that the glycosides act directly on the cell rather than on the insulin molecule. The latter observation is consistent with the ability of those glycosides that act like insulin to do so independent of the hormone. In view of the known insulin-like properties of Con A, the effects of the glycosides seen in the present study suggest roles for a membrane carbohydrate and a carbohydrate binding site in the mechanisms of action of both insulin and Con A. In addition to various alternative explanations, a working hypothesis is presented to rationalize the present observations. It proposes that the effects of the exogenously added glycosides (and Con A) may reflect the presence on the membrane of a native carbohydrate moiety by either mimicking or competitively inhibiting its ability to interact reversibly with a lectin-like carbohydrate binding site associated with the function of the insulin receptor.20

Adipose Tissue

Triglyceride, free fatty acid and carbohydrate metabolism in hyperlipaemic (type IV) and normolipaemic subjects on carbohydrate- of fat-rich diets.

1. Five patients with hyperlipoproteinaemia type IV and five normolipaemic controls were maintained on diets containing either a fat or a carbohydrate content of approximately 60%. Leg exchanges of triglycerides, free fatty acid (NEFA), glucose and other metabolites were measured in the basal state. The metabolic response to an intravenous glucose load and the turnover of intravenously infused (14C)oleic acid were determined. Individual fatty acids were analysed in plasma NEFA and very-low-density lipoprotein triglyceride. 2. No differences were detected between patients and controls with regard to leg uptake of triglyceride or NEFA on either diet. On shifting from a fat-rich to a carbohydrate-rich diet, the control subjects showed reduced values for arterial NEFA and for NEFA turnover, accompanied by a lowered release of NEFA and glycerol from the leg. In patients, on the other hand, the arterial concentration and turnover of NEFA remained unchanged and the release of glycerol from the leg increased. Glucose administration gave a less-pronounced fall of arterial NEFA on either diet in the patients and the incorporation of (14C)oleic acid into triglycerides after glucose administration was more marked than in the controls. 3. No difference in net uptake of triglyceride by the leg was demonstrable between patients and controls. 4. it is concluded that hyperlipaemic patients show an altered metabolic response to a carbohydrate-rich diet, possibly contributing to the development of a hyperlipaemic state, and that acute glucose loading results in a greater incorporation of circulating NEFA into very-low-density lipoproteins in these patients.

Adult

Effects of carbohydrate-free diets on the insulin-carbohydrate relationships in rats.

Insulin-carbohydrate relationships were investigated in four groups of young rats fed low protein diets differing in carbohydrate and fat contents: (1) a diet in which the nonprotein energy was provided by fatty acids (FA); (2) a similar diet in which the fatty acids were substituted by neutral fat (NF); (3) FA diet supplemented with glycerol (FA-Glyc); and (4) a carbohydrate-rich diet (HC). Control rats were fed a stock diet. Rats fed the FA diet lost weight, were hypoglycemic and hypoinsulinemic in the fed state and normoglycemic and normoinsulinemic in the fasted state, and had an impaired glucose tolerance and hyperinsulinemia after a glucose load. Liver and muscle glycogen were low in fed rats. Fasting increased glycogen in liver and decreased glycogen in muscle. NF animals gained weight, were hypoglycemic in both fed and fasted states, and their plasma glucose level after an oral glucose load was almost normal. Plasma insulin/glucose ratio, both in fed and fasted states and after a glucose load indicated hyperinsulinism, which was accompanied by obesity. Muscle and liver glycogen were low in fed animals and did not change after a fast. Supplementation of the FA diet with glycerol (FA-Glyc) abolished weight loss and fasting hyperglycemia and normalized plasma glucose and insulin response to a glucose load. Rats fed the HC diet had an improved glucose tolerance and an increased sensitivity to insulin. Liver glycogen was high in the fed state and normal in the fasted state, whereas muscle glycogen was normal in both nutritional states.

Animals

Carbohydrate metabolism in Agaricus bisporus (Lange) Sing: changes in soluble carbohydrates during growth of mycelium and sporophore.

Changes in the ethanol-soluble carbohydrate content of Agaricus bisporus mycelium and sporophores grown on semi-defined media and commercial compost were studied. The accumulation of mannitol in the sporophore during its growth was not accompanied by an increase in mycelial mannitol. The other major soluble carbohydrate of the sporophore, trehalose, decreased throughout the growth of the sporophore; a parallel decrease was observed in the mycelium. The main accumulation of mannitol was in the pileus and stipe of the sporophore and was accompanied by a decrease in the soluble protein content of these tissues. Before fruiting, glucose and sucrose were present in the mycelial samples in similar quantities to mannitol, but their levels decreased during fruiting. Small quantities of glucose were present in the sporophore. The results are discussed in relation to the possible functions of the soluble carbohydrates.

Basidiomycota

[Various changes in carbohydrate and lipid metabolism and the state of the islands of Langerhans in female rabbits after long-term feeding with food rich in carbohydrates and ovariectomy].

Experiments were conducted on 21 female rabbits. Prolonged excessive feeding with food rich in readily assimilated carbohydrates (bisquits) started at immature age caused hypertrophy and hyperplasia of the insular apparatus of the pancreas, particularly in ovariectomy. The average blood sugar level on fasting stomach displayed a significant reduction in ovariectomized female rabbits. Some female rabbits displayed signs of "exhaustion" of beta-cells of the islets of the pancreas, with the development of latent diabetes mellitus in one of them. Prolonged excessive feeding with food rich in readily assimilated carbohydrates led to the intensification of atherogenesis, most pronounced in the animals with signs of "exhaustion" of beta-cells of the islets of the pancreas.

Animals

Prospective comparison of modified fat-high-carbohydrate with standard low-carbohydrate dietary advice in the treatment of diabetes: one year follow-up study.

1. A prospective randomized study of two dietary regimens has been started in newly-diagnosed diabetics to determine their effect on circulating metabolites and on diabetic complications. 2. During the first year of treatment the fasting plasma glucose concentrations on both the low-carbohydrate diet and the high-carbohydrate, modified-fat (MF) diet showed a similar decrease. 3. Plasma cholesterol showed a sustained decrease only in patients recommended a MF diet. Transient changes in plasma triglyceride concentrations occurred in patients on both dietary regimens. 4. Increased plasma cholesterol levels are associated with atheromatous disease which is common in diabetics in Europe and North America. A MF diet may therefore have an advantage in that it lowers the plasma cholesterol as well as being effective in lowering the plasma glucose.

Adult

Effect of carbohydrate restriction and high carbohydrates diets on men with chemical diabetes.

The influence of low carbohydrate (CHO) diets, starvation, and high CHO diets on glucose tolerance tests (GTT) and plasma insulin response of men with chemical diabetes was studied. The GTT and insulin responses of these seven lean diabetic men were unchanged when the carbohydrate content of the diet was reduced from 44 to 20% of calories. After a 48-hr fast a significant deterioration of the GTT was observed in these diabetic men but the percentage change was identical to that reported previously for normal men. Thus these studies indicate that changes in glucose mtes are quite similar to those reported previously for normal men. The fasting plasma glucose values of seven lean and four obese men with chemical diabetes were significantly lower after one week on a 75% CHO diet than values on a 44% CHO diet. The 75% CHO diet also was accompanied by slight improvements in the oral and intravenous GTT and by slightly lower plasma insulin responses. The improvement in glucose metabolism on high CHO diets appears to results from increased insulin sensitivity. Serum triglyceride values were approximately 55% higher on the 75% CHO diet than values on the 44% CHO diet for the 11 men but these differences were not statistically significant. These studies support previous observations and suggest that high CHO diets may be beneficial in the management of certain diabetic patients. However, further studies are required to determine the long-term effects of high CHO diets containing natural foods on the glucose and lipid metabolism of diabetic patients.

Adult

Periodate oxidation analysis of carbohydrates. XIII. Simultaneous gas chromatographic determination of the aldehydes in the periodate oxidation products of non-dialyzable urinary carbohydrate materials as diethyl dithioacetals.

The aldehydes in the periodate oxidation products of non-dialyzable urinary carbohydrate materials were determined simultaneously by gas chromatography of their diethyl dithioacetal derivatives. The yields of aldehydes for normal male urine varied considerably among subjects, but their molar ratios were almost constant. The average values for glyceraldehyde/glyoxal and lactaldehyde/glyoxal molar ratios were 0.41 and 0.27, respectively. The average amount of L-fucose at the non-reducing terminals of carbohydrate chains, as estimated from the yield of lactaldehyde, was about 5 mg/day.

Aldehydes

[Carbohydrate-reduced diet and metabolism: about the influence of a 4-weeks isocaloric fat-rich, carbohydrate-reduced diet on body weight and metabolism (author's transl)].

The influence of a 4-weeks carbohydrate-reduced, fat-rich diet on 9 slightly overweighted men was investigated. Caloric and protein intake remained unchanged. 70% were fat, 20% carbohydrates. Glucose, cholesterol, triglycerids, serumproteins, urea, uric acid, sodium and potassium were measured in rest. At a 60 minutes bicycle ergometer test glucose, triglycerids, free fatty acids and glycerol were registrated before and after 20 and 60 minutes work. The intensity was 70% of the maximal oxygen uptake. These were the most important results: (1) Body weight decreased continuously. One of the causes is an elevated heat production. (2) Serum-protein and -glucose remained unchanged. Cholesterol and triglycerides were reduced. Serum-urea, -sodium and -potassium diminished continually. On the contrary, uric acid raised over the normal range. (3) Work performance was reduced for 20%. (4) Hypoglycemic values did not appear. The oxidation of fat by the working muscles, and fat mobilisation increased by a fat-rich diet.

Adult

The function of carbohydrate moiety and alteration of carbohydrate composition in human alkaline phosphatase isoenzymes.

The relationship between the structure and function of alkaline phosphatase (orthoposphoric monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1) isoenzymes is under investigation in a number of laboratories. The present study deals with the effects of glycosidase digestion on the alkaline phosphatase isoenzymes. Changes in physicochemcial properties, activity, affinity for various lectins and blood group antisera, carbohydrate composition and biological half-life were investigated. The desialylated hepatic enzyme was shown to be more heat labile and more sensitive to protease digestion in the presence of 0.5% sodium dodecyl sulfate than native hepatic enzyme. Helix contents of the native and desialated hepatic enzyme were calculated to be 39.0 and 30.8%, respectively, and apparent molecular weights 175,000 and 167,000, respectively. Intestinal enzyme preparations treated with alpha-mannosidase, exo-N-acetyl-Dglucosaminidase and endo-N-acetyl-D-glucosaminidase-D displayed a decrease in enzyme activity. Among these, the alpha-mannosidase-treated enzyme activity was the most clearly reduction. The maximum activity of the alpha-mannosidase-treated intestinal enzyme was observed to change from 40 mM Mg2+ to 5--10 mM Mg2+.

Alkaline Phosphatase