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Enhancement of calcium absorption in rats by coadministration of glucose polymer.

Glucose polymer derived from corn starch is widely used in infant formula and nutritional supplements as a readily digestible, low osmolarity source of calories. We have previously observed that glucose polymer causes a marked increase in intestinal calcium absorption in patients with intestinal malabsorption and in normal subjects. The present study investigates the effect of concurrently administered glucose polymer on intestinal 45calcium absorption in rats. The effect of glucose polymer on calcium absorption was compared to that of equivalent doses of dextrose or lactose. Femur radioactivity was determined as an index of calcium absorption. Carbohydrates were prepared at doses of 0.5, 2.0 and 4.0 mg/g body weight and administered with 45Ca by stomach gavage. Control rats received 45Ca in water alone. Coadministration of glucose polymer resulted in a dose-dependent increase in calcium absorption. At the highest dose of polymer studied calcium absorption was 75% greater than in control animals. Administration of lactose and dextrose also resulted in a dose-dependent increase in calcium absorption. Calcium absorption was 122 and 67% greater than controls when 4 mg/g BW lactose and dextrose were given with the 45Ca. We conclude that glucose polymer stimulates calcium absorption in rats similar to lactose and glucose. These results suggest that glucose polymer may be a useful therapeutic adjunct in patient groups in which there is a desire to increase intestinal calcium absorption.

Animals↗

Transient glucosylation of protein-bound Man9GlcNAc2, Man8GlcNAc2, and Man7GlcNAc2 in calf thyroid cells. A possible recognition signal in the processing of glycoproteins.

Calf thyroid slices incubated with [U-14C]glucose synthesized protein-bound Glc3Man9GlcNAc2, Glc2-Man9GlcNAc2, Glc1Man9GlcNAc2, Glc1Man8GlcNAc2, and Glc1Man7GlcNAc2. Although label in the glucose residues of the last three compounds could be detected within 5 min of incubation, appearance of radioactivity in the mannose residues of the alpha-mannosidase-resistant cores of Glc1Man8GlcNAc2 and Glc1Man7GlcNAc2 took more than 30 and 60 min, respectively, to appear after label was detected in the same mannose residues of Glc1Man9GlcNAc2. The glucose residues were removed upon chasing the slices with unlabeled glucose. The last compound to disappear was Glc1Man9GlcNAc2. Calf thyroid microsomes incubated with UDP-[U-14C]Glc synthesized the five protein-bound oligosaccharides mentioned above. Although addition to GDP-Man to the incubation mixtures greatly diminished the formation of Glc3Man9GlcNAc2 bound either to dolichol-P-P or to protein, labeling of Glc1Man9GlcNAc2, Glc1Man8GlcNAc2, and Glc1Man7GlcNAc2 was not affected. Addition of kojibiose prevented deglucosylation of protein-bound Glc3Man9GlcNAc2 without affecting the formation of Glc1Man8GlcNAc2 and Glc1Man7GlcNAc2 and only partially diminishing that of Glc1Man9GlcNAc2. These results indicate that Glc1Man8GlcNAc2 and Glc1Man7GlcNAc2 were formed by glucosylation of the unglucosylated species and not be demannosylation of Glc1Man9GlcNAc2 and that probably part of the latter compound was formed in the same way.

Animals↗

PATTERNS OF OXIDATIVE ASSIMILATION IN STRAINS OF PSEUDOMONAS AND ACHROMOBACTER.

Tomlinson, Geraldine A. (University of British Columbia, Vancouver, B.C., Canada) and J. J. R. Campbell. Patterns of oxidative assimilation in strains of Pseudomonas and Achromobacter. J. Bacteriol. 86:434-444. 1963.-Oxidative assimilation of glucose-U-C(14) in the absence of added nitrogen was studied by use of washed-cell suspensions of Pseudomonas aeruginosa, P. fluorescens, Achromobacter strain B81, and Achromobacter viscosus (Alcaligenes viscolactis). The suggestion that oxidative assimilation in these organisms is the reincorporation of endogenously produced ammonia by way of alpha-ketoglutarate is tenable. Each of the four organisms accumulated intermediate compounds which acted as pacemakers for the oxidation of glucose. This phenomenon, partly because it ensured the availability of additional ammonia, undoubtedly increased the degree of oxidative assimilation. Products accumulating in the supernatant fluids during glucose oxidation were alpha-ketoglutarate, pyruvate, gluconate, a low molecular weight carbohydrate, and dicarboxylic acids. No two bacteria formed the same products. Assimilation of radioactivity into the cells, which accounted for 12 to 26% of the available C(14), continued as long as an oxidizable substrate was present, and was paralleled by uptake of endogenously produced ammonia. During the early stages of glucose oxidation, compounds of the cold trichloroacetic acid-soluble pool constituted a major portion of the total radioactivity of the cells. The lipid fractions of P. aeruginosa and Achromobacter B81 were also of high relative activity during this time. The labeling of the nucleic acid fractions of all four bacteria increased with time, more radioactivity being found in fractions from the two Achromobacter species than in those from the pseudomonads. At the completion of the experiment, the largest percentage of incorporated radioactivity was present in the protein fractions. One of the organisms, Achromobacter B81, synthesized a high molecular weight carbohydrate material.

Achromobacter↗

Identification and characterization of the glucose-transport protein of the bovine blood/brain barrier.

The glucose-transport protein from bovine cerebral-cortex microvessels has been identified and characterized by virtue of its ability to bind the ligand [4-3H]cytochalasin B. Microvessel membranes were found to contain a single set of glucose-inhibitable high-affinity cytochalasin B-binding sites [113 +/- 16 (S.E.M.) pmol/mg of membrane protein], with an association constant of 6.8 +/- 1.8 (S.E.M.) micron-1. D-Glucose inhibited the binding to these sites with a Ki of 31 mM. The transport protein was identified by photoaffinity labelling with [4-3H]cytochalasin B and was found to migrate as a broad band of apparent Mr 55,000 on SDS/polyacrylamide gels. Labelling was inhibited by D-glucose, but not by L-glucose. Treatment with endoglycosidase F yielded a sharper band of apparent Mr 46,000, indicating that the transport protein is glycosylated. However, in contrast with the human erythrocyte glucose transporter, digestion with endo-beta-galactosidase had little effect on the electrophoretic mobility of the microvessel protein. Tryptic digestion of the photolabelled protein yielded a radioactive fragment of apparent Mr 18,000, similar to that of the fragment produced by digestion of the labelled human erythrocyte glucose transporter. In addition, a protein of Mr identical with that of the photolabelled transporter was labelled on Western blots of microvessel membranes by antisera raised against the intact erythrocyte transporter and against synthetic peptides corresponding to its N- and C-terminal regions. It is concluded that the glucose-transport protein of bovine cerebral-cortex microvessel endothelial cells shows structural homology with the human erythrocyte glucose transporter.

Animals↗

Use of active site-directed inhibitors to study in situ degradation of glycoproteins by the perfused rat liver.

Use was made of the asialoglycoprotein receptor system in a perfused rat liver in order to study lysosomal degradation and subsequent metabolism of radioactive derivatives of asialo-ovine submaxillary mucin and asialo-alpha 1-acid glycoprotein. A trace of N-acetyl-D-[6-3H]galactosamine-labeled asialo-ovine submaxillary (4 micrograms) was completely taken up by the tissue in less than 20 min. After 3 h 24% of the radioactivity from the mucin reappeared on newly synthesized serum glycoproteins that were secreted into the perfusate. [6-3H] Galactose asialo-alpha 1-acid glycoprotein was also rapidly cleared by the liver; however, after 3 h greater than 60% of the radioactivity derived from this sugar labeled glycoprotein was secreted back into the perfusate as [3H]glucose. Rat livers perfused with 0.15 mM beta-D-galactopyranosylmethyl-p-nitrophenyltriazene lost 90% of their beta-D-galactosidase activity within 1 h while other representative glycosidases showed no change as followed by hydrolysis of p-nitrophenylglycosides. Livers pretreated with this triazene compound metabolized [3H]GalNAc asialo-ovine submaxillary mucin normally but were unable to process [3H]Gal asialo-alpha 1-acid glycoprotein as evidenced by a complete inhibition of [3H]glucose release following addition of the latter substrate. Metabolism of N-acetyl[14C]glucosamine asialo-alpha 1-acid glycoprotein was similarly inhibited by 70%. 125I-labeled asialo-alpha 1-acid glycoprotein catabolism was not affected by the chemically induced beta-D-galactosidase deficiency. Subcellular fractionation of inhibitor-treated livers accumulating radioactive carbohydrate showed the majority of the label was associated with a fraction enriched in lysosomes. Analysis of the trapped radioactivity by high resolution Bio-Gel P-4 chromatography revealed nearly intact oligosaccharides minus only the reducing N-acetylglucosamine of the chitobiose core. Direct comparison of these sugar chains with those isolated from human and canine GM1 gangliosidosis liver by silicic acid thin layer chromatography showed those isolated from rat liver to be identical to the major subset of oligosaccharides found in the human disease. In similar experiments in which the galactosyl triazene was replaced by swainsonine, an alpha-D-mannosidase inhibitor, catabolism of [14C]GlcNAc asialo-alpha 1-acid glycoprotein resulted in the accumulation of a single oligosaccharide of the structure. Man3[14C]GlcNAc1. These results demonstrate an endo-N-acetyl-beta-D-glucosaminidase is active in rat liver lysosomes.

Acetylgalactosamine↗

Serine synthesis by an isolated perfused rat kidney preparation.

The isolated perfused rat kidney was shown to synthesize serine from aspartate or glutamate, both of which are also precursors of glucose. The major products of aspartate metabolism were ammonia, serine, glutamate, glucose, glutamine and CO2. Perfusion of kidneys with aspartate in the presence of amino-oxyacetate resulted in a near-complete inhibition of aspartate metabolism, illustrating the essential role of aspartate aminotransferase in the metabolism of this substrate. Radioactivity from 14C-labelled aspartate and from 14C-labelled glycerol was incorporated into serine and glucose. Production of both glucose and serine from aspartate was suppressed in the presence of 3-mercaptopicolinic acid. These data provide evidence for the operation of the phosphorylated and/or non-phosphorylated pathway for serine production to the presence of 3-mercaptopicolinic acid. This is explained by simultaneous glycolysis. The rate of glucose production, but not that of serine, was greater in kidneys perfused with glutamate or with aspartate plus glycerol than the rates obtained by perfusion with aspartate alone. These data are taken to suggest that serine synthesis occurred at a near-maximal rate, and that the capacity of the kidney for serine synthesis from glucose precursors is lower than that for glucose synthesis.

Amino Acids↗

Glucose-C14 metabolism of dormant and activated ascospores of Neurospora.

Budd, Kenneth (The University of Michigan, Ann Arbor), Alfred S. Sussman, and Frederick I. Eilers. Glucose-C(14) metabolism of dormant and activated ascospores of Neurospora. J. Bacteriol. 91:551-561. 1966.-Dormant and activated ascospores of Neurospora tetrasperma, incubated in C(14)-labeled glucose, absorb and metabolize this sugar. At the same time, up to 55% of the CO(2) production from endogenous substrates is quenched, whereas total CO(2) production is unchanged. Glucose-carbon appears in CO(2), lipids, and ethyl alcohol-soluble and -insoluble material in both dormant and activated ascospores, although the proportions entering these fractions differ in the two groups of spores. With few exceptions, the identifiable intermediates of glucose metabolism are the same in dormant and activated ascospores, indicating that the principal pathways may be identical. During glucose metabolism, dormant ascospores accumulate a nondialyzable, ethyl alcohol-soluble polymer, or polymers, which is either absent from activated spores or present in much smaller amounts. This material contains glucose, ribose, and at least nine amino acids, and may represent precursors of more complex cell material which accumulate because of an enzymatic deficiency in the dormant spore. Radioactivity is incorporated into all fractions of the dormant spores and into CO(2) without a noticeable lag, indicating that most, if not all, of the enzymes for glucose utilization are present. A lag in incorporation is observed in the activated spores, which most probably is due to rapid endogenous production of glucose from trehalose, resulting in dilution of lable. After absorption of labeled glucose, two pools of trehalose are found in dormant spores, one of which is extractable without breaking the spores, and the other, only after the spores are disintegrated. The widely differing specific radioactivity of the two pools indicates that these are separated in the intact spore.

Carbon Dioxide↗

The deoxyglucose method in the ferret brain. I. Methodological considerations.

In the brain of the anesthetized ferret, the 2-deoxyglucose (2-DG) transfer rate constants required to determine cerebral glucose utilization by the deoxyglucose method were calculated from regional gray matter time-radioactivity curves measured for 180 min after tracer injection. Results suggest that loss of metabolized tracer from brain occurs at a rate of about 1%/min for the first 180 min after injection if the rate constant of the rate-limiting step for loss of metabolized tracer (k4*) represents a first-order kinetic process. A simulation experiment shows that, whether k4* is assumed to be 0 or 0.01 min-1, has a negligible influence on glucose utilization rates obtained in conventional 45 min autoradiographic experiments provided that the entire analysis, including lumped constant determination, is carried out in a consistent way. The 2-DG lumped constant for k4* = 0 is 0.54, and 0.68 for k4* = 0.01 min-1.

Animals↗

Radioactive deoxyglucose uptake into forelimb muscles in task performing monkeys.

Incorporation of [14C]2-deoxy-d-glucose (2-DG) into forelimb muscles of task-performing monkeys was studied. Extensor muscles of used forelimbs showed a significant increase in 2-DG uptake; they were ext.dig.2nd and 3rd, ext.dig.com., ext.carp.rad. and ext.carp.uln. On the other hand, flexor muscles in the used forelimb did not show such a significant increase in 2-DG uptake. Muscles of non-used limbs, flexors or extensors, did not show a significant 2-DG uptake.

Animals↗

[Relationship between three neuropsychological tests and cerebral glucose metabolism in Alzheimer's disease].

OBJECTIVE: To study the relationship between the scores of 3 neuropsychological scales: MMSE, extended scale of dementia (ESD) and Blessed dementia scale (BDS) and cerebral glucose metabolism in Alzheimer's disease (AD) indicated by positron emission tomography (PET). METHODS: Brain scanning was performed with SIEMENS ECAT 47 PET scanner among 21 AD patients, aged 60 - 83 (10 mild, 6 moderate, and 5 severe), and 15 healthy persons, aged 57 - 73. The ratio of mean radioactivity of cerebral lobe to that of cerebellum as semi-quantitative parameters was used to evaluate the cerebral glucose metabolism. Cognitive function was assessed by three neuropsycholigical scales: MMSE, ESD and BDS. RESULTS: The average MMSE score was 13.3 +/- 6.3 (range 2 - 25), the average ESD score was 120.5 +/- 54.8 (range 28 - 200), and the average BDS score was 4.7 +/- 3.1 (range 0.5 - 10) in the AD patients. The decreases of glucose metabolism in parietal, frontal and temporal lobes were significantly positively correlated with MMSE and ESD scores, and negatively correlated with BDS scores in AD patients (P < 0.05). The correlations between MMSE and ESD (r = 0.886), MMSE and BDS (r = -0.763), and ESD and BDS (r = -0.773) were significant in AD patients (all P < 0.01). Taking the ratio of radioactivity of cerebral lobe to that of cerebellum as an independent variable X, the MMSE, ESD and BDS scorsas dependent variable Y, three regression equations were established as follows: Y = 40.11 * X- 25.32, Y = 309.19 * X- 180.9, Y = 19.97 - 16.53 * X. The independent variable entering the three regression equations was always the ratio of radioactivity of left parietal lobe to that of cerebellum. The ratio of radioactivity of left parietal lobe to that of cerebellum was a significant predictor for cognitive dysfunction measured by MMSE, ESD and BDS in AD patients (P < 0.01). CONCLUSION: MMSE, ESD and BDS scores correlate well with impaired cerebral glucose metabolism in AD. The function of left parietal lobe may play an important role in the progress of the disease. MMSE, ESD and BDS are good measures for cognitive dysfunction.

Aged↗

Importance of local versus systemic effects of non-steroidal anti-inflammatory drugs in increasing small intestinal permeability in man.

Increased small intestinal permeability caused by non-steroidal anti-inflammatory drugs (NSAIDs) is probably a prerequisite for NSAID enteropathy, a source of morbidity in patients with rheumatoid arthritis. This increased small intestinal permeability may be a summation of a local effect during drug absorption, a systemic effect after absorption, and a local effect of the drug excreted in bile, but the relative contribution made by these factors is unknown. We assessed the effect of indomethacin and nabumetone on intestinal permeability. The principal active metabolite of nabumetone, 6-methoxy-2-naphthylacetic acid, is not subject to appreciable enterohepatic recirculation. Twelve volunteers were studied before and after one week's ingestion of indomethacin (150 mg/day) and nabumetone (1 g/day) with a combined absorption/permeability test. Neither drug had a significant effect on the permeation of 3-0-methyl-D-glucose, D-xylose, and L-rhamnose. Indomethacin increased the permeation of radioactive 51chromium ethylenediaminetetra-acetic acid (51Cr EDTA) significantly from baseline (mean (SEM) 0.63 (0.09)% v 1.20 (0.14)%, p less than 0.01) but nabumetone did not (0.70 (0.10)% p greater than 0.1). These results were supported by the 51Cr EDTA/L-rhamnose urine excretion ratios, which reflect changes in intestinal permeability. They suggest that NSAIDs increase intestinal permeability during absorption or after biliary excretion and that the systemic effect is of minor importance.

Adult↗

Clinical applications of assessments of myocardial substrate utilization with positron emission tomography.

Positron emission tomography (PET) permits sequential, noninvasive assessment of myocardial perfusion and metabolism. Based on the pattern of substrate use, clinical studies have utilized PET to define the location and extent of myocardial infarction, to identify areas of jeopardized but viable myocardium, and to assess the metabolic response of the myocardium to pharmacological therapy as well as to interventions such as coronary thrombolysis and coronary artery bypass surgery. The ability to noninvasively assess specific metabolic pathways should facilitate our understanding of normal myocardial metabolism, its perturbations with cardiac disease, and thereby improve the diagnosis and treatment of the biochemical processes underlying cardiac dysfunction.

Amino Acids↗

125I-glucagon-degrading activity in acid-saline extracts of rat salivary gland.

The antibody-binding ability of the glucagon-like substance in rat submaxillary gland acid saline extract was examined by affinity chromatography, and the biological activity studied using the isolated liver perfusion method. We found that the glucagon-like substances in acid saline extract could not be bound to anti-glucagon antibody and that the gel-filtration peak on ultrogel AcA 54 could increase neither glucose nor cyclic AMP output from isolated perfused rat liver. Furthermore, the radioactivity peak of 125I-glucagon on Bio Gel P-6 column chromatography moved from its original position and eluted in later fractions after incubation with an acid saline extract of the submaxillary gland. In consequence, there was 125I-glucagon degrading activity in the submaxillary gland, but no glucagon-related peptide. Therefore, it is suggested that the glucagon-like substance, which has been reported in acid saline extract of the rat salivary gland, may be an artifact due to tracer degrading activity.

Animals↗

Acute effects of ethanol in the control of protein synthesis in isolated rat liver cells.

The acute effect of ethanol on hepatic protein synthesis is a rather controversial issue. In view of the conflicting reports on this subject, the effect of ethanol on protein labeling from L-[3H]valine in isolated liver cells was studied under a variety of experimental conditions. When tracer doses of the isotope were utilized, ethanol consistently decreased the rate of protein labeling, regardless of the metabolic conditions of the cells. This inhibition was not prevented by doses of 4-methylpyrazole large enough to abolish all the characteristic metabolic effects of ethanol, and it was not related to perturbations on the rates of L-valine transport and/or proteolysis. When ethanol was tested in the presence of saturating doses of L-[3H]valine no effect on protein labeling was observed. These observations suggest that the ethanol effect in decreasing protein labelling from tracer doses of the radioactive precursor does not reflect variations in the rate of protein synthesis but reflects changes in the specific activity of the precursor. These changes probably are secondary to variations in the dimensions of the amino acid pool utilized for protein synthesis. Even though it showed a lack of effect when tested alone, in the presence of saturating doses of the radioactive precursor ethanol inhibited the stimulatory effects on protein synthesis mediated by glucose and several gluconeogenic substrates. This effect of ethanol was not prevented by inhibitors of alcohol dehydrogenase, indicating that a shift of the NAD system to a more reduced state is not the mediator of its action. It is suggested that ethanol probably acted by changing the steady-state levels of some common effector(s) generated from the metabolism of all these fuels or else by preventing the inactivation of a translational repressor.

Animals↗

Quantitation of the pathways followed in the conversion of fructose to glucose in liver.

Recently, only about 50% of the conversion of fructose to glucose was reported to be via fructose-1-P aldolase catalysis in children. This was also suggested to be the case in adults. That possibility has been tested using a method that quantifies the pathways of fructose conversion to glucose via the fate of 14C from specifically labeled fructose. Trace [6-14C] fructose or its immediate precursor [6-14C]sorbitol with unlabeled fructose (0.3 mg/kg body weight/min) was given intravenously or intragastrically with trace [1-14C]lactate to six normal adults fasted overnight. The distributions of 14C in glucose from blood samples were determined. The ratios of 14C in C1 to C6 of the glucose were equal to or only slightly less than the ratios of 14C in C3 to C4. Since incorporation into C3 and C4 of glucose must have arisen via the conversion of [1-14C]lactate to [1-14C]triose phosphates, fructose conversion to glucose must also have arisen predominantly via the triose phosphates. From the ratios, 85.1% to 100%, a mean of 94.9% of the fructose converted to glucose is calculated to have been converted to glucose with cleavage of the carbon skeleton of the fructose. These findings contrast with the report that in children under similar conditions only about 50% of the conversion of fructose to glucose is with cleavage. The findings agree with previous results in which fructose was administered to normal adults as a bolus at a dose of 60 mg/kg body weight. The possible reasons that the findings in children are different from those in adults are considered.

Adult↗

Influence of glucose and urea on 125I transport across an anion exchange paper membrane.

In order to study the influence of glucose and urea on the 125I transport across an anion exchange paper membrane, the transmembrane potential, the fluxes, and the concentrations of 125I, glucose and urea within the membrane were measured in the Na125I concentration-cell system containing glucose or urea. Glucose and urea increased the membrane/solution distribution of the iodide ion, but scarcely affected the diffusion process of iodide ion within the membrane.

Glucose↗

[Significance of PET for ENT-tumors].

Significance for ENT-tumors. Molecular imaging with PET is based on the use of specific radioactive molecules as source of image contrast. In ENT-tumors mostly glucose and occasionally amino acid/protein metabolism were assessed with PET for tumor diagnosis. Thymidine salvage pathway and proliferation as well as tissue hypoxia are tested already in clinical studies and bear considerable potential for modulation of radiation ports and therapeutic response of cytoreductive regimens. This short review summarises actual clinical indications and potential of PET in ENT-tumors. For both nodal staging as well as detection of recurrent disease, sensitivity and specifity of FDG-PET were 80 - 100 %. FDG-PET proved to be superior to conventional imaging in most published studies. In CUP-syndrome the primary could be detected in 25 - 50 % of patients. Acquisition of PET and CT images in a combined scanner allow versatile PET based metabolic imaging in combination with high resolution and anatomical precise CT-based morphological imaging and thus combines advantages of both imaging modalities. Clinical as well as scientific potential of this functional metabolic and high resolution morphological imaging approach is high.

Amino Acids↗