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PubMed1989

Alkaline phosphatase cDNA transfected cells promote calcium and phosphate deposition.

The full length cDNA of rat alkaline phosphatase (AP) was placed under the control of the SV40 early promoter. This plasmid was transfected by the calcium phosphate method into AP negative ROS 25/1 cells. Ten clones with AP specific activities ranging between 0.1-2 mumole/min/mg were isolated by cotransfection with the plasmid pSV2Neo, which renders the cells resistant to the antibiotic G418. Two clones with different AP specific activities: C (0.01 mumole/min/mg) and S (2.0 mumole/min/mg) and the osteoblastic ROS 17/2.8 cells (2.0 mumole/min/mg), were examined for their ability to mineralize. In vitro mineralization was tested by culturing cells in alpha-MEM containing 10% fetal bovine serum and 50 micrograms/ml ascorbate in the presence or absence of 10 mM beta-glycerophosphate. Mineralized deposits were observed in all cultures of the S clone and ROS 17/2.8 cells in the presence of beta-glycerophosphate, but not in C clones. Measurement of calcium and phosphorus levels in cells correlated with AP levels of transfected cells. However, extent of mineral accumulation in the transfected ROS 25/1 cells differ from the osteogenic ROS 17/2.8 cells. This finding indicates that high levels of AP may be a necessary constituent for the mineralization process together with other factors yet to be identified.

Alkaline Phosphatase↗

PubMed1979

Some lipogenic enzyme activities in rat livers in which an excessive fat accumulation occurred due to feeding low-level amino acid mixture diets.

In liver homogenates of rats fed a low-level diet of Wheat-Rice- or Miyazaki-pattern amino acid mixture, some enzymes such as glucose-6-phosphate dehydrogenase, ATP citrate lyase, fatty acid-synthesizing enzymes, malic enzyme and L-alpha-glycerophosphate dehydrogenase, whose activities are indicators of lipogenesis have been determined from the viewpoint of the mechanisms producing fatty liver. In the early experimental period, malic enzyme activity increased more markedly in rats fed low amino acid mixture diets than in the control group, and L-alpha-glycerophosphate dehydrogenase activity in the liver increased slightly. Conversely, ATP citrate lyase and fatty acid-synthesizing enzyme activities remained almost at control levels, or glucose-6-phosphate dehydrogenase activity tended to decrease. These results suggest that some other associated factors, such as depression of the lipid transfer system in the liver rather than accelerated lipogenesis itself, may be the main cause of the fatty livers produced under these nutritional conditions.

ATP Citrate (pro-S)-Lyase↗

PubMed1999

Effect of dietary taurine supplementation on GSH and NAD(P)-redox status, lipid peroxidation, and energy metabolism in diabetic precataractous lens.

PURPOSE: To evaluate changes in glutathione and NAD(P)-redox status, taurine and malondialdehyde (MDA) levels, glucose utilization, and energy metabolism in diabetic precataractous lenses and to assess whether these changes can be prevented with dietary taurine supplementation. METHODS: The experimental groups included control and streptozotocin-diabetic rats with a 3-week duration of diabetes fed unsupplemented or taurine (1% or 5%)-supplemented diets. The levels of glucose, sorbitol, fructose, myo-inositol, oxidized glutathione (GSSG), glycolytic intermediates, malate, alpha-glycerophosphate, and adenine nucleotides were assayed in individual lenses spectrofluorometrically by enzymatic methods, reduced glutathione (GSH) spectrofluorometrically with O-phthaldialdehyde, MDA colorimetrically with N-methyl-2-phenylindole, and taurine by high-performance liquid chromatography. Free cytosolic NAD+/NADH and NADP+/NADPH ratios were calculated from the lactate dehydrogenase and malic enzyme systems. RESULTS: Sorbitol pathway metabolites and MDA were increased, and GSH and taurine levels were reduced in diabetic rats versus controls. The profile of glycolytic intermediates (an increase in glucose 6-phosphate, no change in fructose 6-phosphate and fructose 1,6-diphosphate, an increase in dihydroxyacetone phosphate, a decrease in 3-phosphoglycerate, phosphoenolpyruvate, and pyruvate, and no change in lactate), and a 9.2-fold increase in alpha-glycerophosphate suggest diabetes-induced inhibition of glycolysis. Free cytosolic NAD+/NADH ratios, ATP levels, ATP/ADP, and adenylate charge were reduced, whereas free cytosolic NADP+/NADPH ratios were elevated. Lens taurine levels in diabetic rats were not affected by supplementation with 1% taurine. With 5% taurine supplementation, they were increased approximately 2.2-fold higher than those in untreated diabetics but remained 3.4-fold lower than in controls. Lens GSH levels were similar in diabetic rats fed unsupplemented and 5% taurine-supplemented diets, whereas GSSG and MDA levels and GSSG/GSH ratios were reduced by 5% taurine supplementation. The decrease in free cytosolic NAD+/NADH, ATP/ADP, and adenylate energy charge were ameliorated by 5% taurine supplementation, whereas accumulation of sorbitol pathway intermediates, depletion of myoinositol, inhibition of glycolysis, a decrease in ATP and total adenine nucleotide, and an increase in free cytosolic NADP+/NADPH were not prevented. CONCLUSIONS: Dietary taurine supplementation ameliorates MDA levels, GSSG/GSH, and NAD+/NADH and fails to prevent the osmotically mediated depletion of GSH and taurine and the decrease in glucose utilization and ATP levels in diabetic precataractous lens. Dietary taurine supplementation cannot be regarded as an alternative to aldose reductase inhibition in eliminating antioxidant and metabolic deficits contributing to diabetes-associated cataractogenesis.

Adenosine Triphosphate↗

PubMed1999

[The effect of immobilized salivary alkaline phosphatase on remineralization processes].

Relationship between alkaline phosphatase immobilized on apatite and remineralization processes has been studied. The enzyme notably accelerated the remineralization. The course of enzymatic remineralization depended on the source of phosphorus (inorganic phosphorus or beta-glycerophosphate). Calcium incorporation in remineralizing surface was the most rapid in the presence of beta-glycerophosphate. This may be explained by a higher rate of formation of apatite under the effect of immobilized alkaline phosphatase, than of brushite or vitlokite.

Alkaline Phosphatase↗

PubMedDate not supplied

[The carbohydrate-containing cell wall polymers of some species from the cluster "Streptomyces lavendulae"].

The type strains of the cluster "Streptomyces lavendulae" species with a low level of DNA-DNA relatedness were found to contain different cell-wall carbohydrate polymers, whereas the species of this cluster with a level of DNA-DNA relatedness of about 60% contain similar or identical carbohydrate polymers. The type strains Streptomyces katrae VKM Ac-1220T and S. polychromogenes VKM Ac-1207T synthesize mannan with different amounts of alpha-1,2- and alpha-1,3-substituted mannopyranose units and a small number of 1,3-poly(glycerophosphate) chains. The cell walls of S. lavendulocolor VKM Ac-215T and Streptomyces sp. VKM Ac-2117 were found to contain a hitherto unknown teichuronic acid, whose repeating unit is a disaccharide consisting of diaminomannuronic acid and N-acetylgalactosamine:-->4)-beta-D-ManpNAc3NAcA-(1-->3)-alpha-D- GalpNAc-(1-->. In addition, the cell walls of these two streptomycetes contain beta-glucosylated 1,5-poly(ribitol phosphate). The cell walls of S. virginiae VKM Ac-1218T and S. flavotricini VKM Ac-1277T contain the same poly(glucosyl-glycerophosphate). The results presented in this paper are in accordance with the DNA-DNA relatedness data published earlier and indicate a taxonomic significance of the structure of the cell-wall polysaccharides for the delineation of pheneticall/related Streptomyces species.

Cell Wall↗

PubMedDate not supplied

[New strains of soil bacilli which mineralize organic phosphorus compounds].

Microorganisms, which mineralize organophosphates (216 isolates) have been isolated for the chernozem soils of different regions of Ukraine. Sixteen most active strains referred to Bacillus genus have been identified and studied. These microorganisms grow (with different intensity) on the mineral medium with glucose and glycerophosphate or phytin, accumulating phosphate-ions in the medium. The most active strains mineralized glycerophosphate, accumulating about 280 micrograms/ml of PO4(3-) and phytin accumulating up to 130 micrograms/ml of PO4(3-) during 3 days. New strains of bacilli have been chosen for creation of combined bacterial preparations of the complex effect.

Bacillus↗

PubMed2001

Molecular species composition of rat liver phospholipids by ESI-MS/MS: the effect of chromatography.

Using electrospray ionization tandem mass spectrometry (ESI-MS/MS) this study shows that the loss of glycerophospholipid (GPL) after chromatography was unevenly distributed across the GPL molecular species. Both TLC and HPLC caused a preferential loss of GPL with 0 to 3 double bonds: 20% and 7.2% for choline glycerophosphates (PC) and 19.7% and 7.5% for ethanolamine glycerophosphates (PE), respectively. A consequence of these losses was that GPLs containing fatty acids with four or more double bonds had a greater contribution to the total after chromatography. ESI-MS/MS analysis also showed that PC molecular species with four or more double bonds migrated at the front of the TLC band of PCs. GPLs extracted from TLC plates occasionally contained PCs that were smaller than those in the original extract. These low molecular mass PCs were easily reduced to alcohols and formed derivatives with 2,4-dinitrophenylhydrazine, suggesting that aldehydes were generated by the oxidation of unsaturated fatty acids. Directly analyzing lipid extracts by ESI-MS/MS without preliminary chromatographic separation gives an accurate distribution of GPL molecular species in lipid mixtures. However, the ionization of the phospholipids in the electrospray jet maximized at relatively low concentrations of GPL. There was a linear response between phospholipid mass and ion intensity for concentrations around 1-2 nmol/ml for both PC and PE. The total ion intensity continued to increase with concentrations above 1-2 nmol/ml, but the response was non-linear.

Animals↗

PubMed2001

[Assembly and evaluation of tissue engineered human cellular alveolar bony implanting materials in vitro].

OBJECTIVE: To assembly human cellular alveolar bony implanting materials in vitro and evaluate their osteogenic activities. METHODS: Human alveolar bone cells were separated from alveolar bone around the third impacted teeth of 3 patients by enzyme digestion and cultured in alpha-MEM containing beta-glycerophosphate and Dexamethasone at 5% CO2, 37 degrees C for 21-28 days. Confirmed osteoblasts-like cells were then seeded onto 3 kinds of degradable biomaterials of polyglycolic acid scaffold, collagen sponge, and L-lactic acid/epsilon-caprolactone to form the cell-scaffold complexes. The 3 types of complexes were continued to culture for 21-28 days at the same conditions. The cell proliferation, morphological changes, ALPase activity and mineral nodule formation on scaffolds were measured and observed at 3 days intervals. RESULTS: The results indicated that the cultured human alveolar bone origin cells from 3 patients could successfully express the osteoblasts phenotype in single layered culturing after stimulated by beta-glycerophosphate and Dexamethasone. The cultured osteoblast-like cells seeded on PGAS matrix had the highest attachment, proliferative and osteogenic activities, suggesting a good bio-affinity between the human alveolar osteoblast-like cells and the PGAS matrix. The statistical analysis (ANOVA) showed that there were significant differences between PGAS- osteoblasts complex and CLGS or LACT complexes on osteogenic activities (P < 0.05). CONCLUSION: PGAS-osteoblast complex is worth to be further developed into a tissue-engineered cellular artificial bony implant for reconstructing the oral-maxillofacial bony defects.

Absorbable Implants↗

PubMedDate not supplied

[Generation and transport systems of restored equivalents in rabbit brain tissue under experimental atherosclerosis].

Distrubances are found in the shuttle transport systems of extramitochonidral hydrogen in the brain of rabbits with experimental atherosclerosis, the catalytical function of glyceraldehyde phosphate dehydrohenase being preserved. A decrease in the activity of cytoplasmatic glycerophosphate dehydrogenase was observed parallel with an increase in the content of glycerophosphate. Deviations from the norm in the malate-aspartate shunt are pronounced in a considerable activation of mitochondrial malate dehydrogenase, inhibition of aspartate amino transpherase, accumulation of oxaloacetate and exhaustion of the malate stock. Beta-Oxybutyrate dehydrogenase activates sharply, the content of acetoacetate increases. The total activity of hexoxokinase in cytoplasm and mitochondria remains practically unchanged.

Animals↗

PubMed1992

[Lymphocyte enzymes, activity of lipid peroxidation processes and the antioxidant protection of patients with tuberculosis of the lungs].

Activity of succinate dehydrogenase, alpha-glycerophosphate dehydrogenase, NAD2- and NADP2-diaphorases and acid phosphatase in lymphocytes of the peripheral blood as well as malonic dialdehyde and alpha-tocopherol, as parameters of lipid peroxidation defense, were studied in 49 patients with different forms of pulmonary tuberculosis and in 17 practically and clinically healthy subjects. Patients with focal pulmonary tuberculosis presented drop of succinate dehydrogenase and NAD2-diaphorase activity and rise of alpha-glycerophosphate dehydrogenase and NADP2-diaphorase activity in lymphocytes. Parameters of malonic dialdehyde and alpha-tocopherol in patients and healthy subjects had no difference. Patients with infiltrative and fibrocavernous pulmonary tuberculosis had drastic suppression of energy enzymes and sharp rise of acid phosphatase activity in lymphocytes, which correlated with a significant rise of malonic dialdehyde level and decrease of blood serum alpha-tocopherol. There was a tendency to an increase in energy enzymes activity and decrease of acid phosphatase activity 3-4 months after chemotherapy, which was followed by the clinical improvement of patients' condition. Direct dependence was found between the normalization of enzyme activity of lymphocytes and diminution of lipid peroxidation processes.

Adult↗

PubMed1992

Ultrastructural localisation of acid phosphatase in intestinal eosinophilic granule cells (EGC) of rainbow trout (Oncorhynchus mykiss) following degranulation with capsaicin.

Enzyme cytochemistry was used to investigate possible lysosome involvement in capsaicin induced degranulation of the eosinophilic granule cell (EGC) of the rainbow trout intestine. Three adult rainbow trout (Oncorhynchus mykiss) were injected intraperitoneally with capsaicin in a saline vehicle (0.5 micrograms.g-1 body weight). Following a 2 hour period of incubation, the fish were killed, and a mid portion of the intestine was dissected and fixed in cold glutaraldehyde buffered with sodium cacodylate. Vibratome sections were incubated in either reaction medium containing beta-glycerophosphate and cerium chloride in acetate buffer or substrate (beta-glycerophosphate) deficient control medium. Sections were then refixed in osmium tetroxide and processed for electron microscopy. Acid phosphatase was found to be localised within lysosomes. The enzyme was not found in the large cytoplasmic granules under normal or capsaicin-stimulated conditions. EGCs which had migrated to the lamina propria in response to the capsaicin stimulation had a distinct multivesicular granule morphology. These multivesicular granules did not contain acid phosphatase suggesting that this form of EGC degranulation is not a lysosomally mediated event.

Acid Phosphatase↗

PubMedDate not supplied

[Effect of cultivation conditions of Bacillus subtilis IMV B-7023 on surface properties of cells].

It has been established that the cells of Bacillus subtilis IMV B-7023 grown on the medium where peptone or calcium glycerophosphate served as the only source of carbon and energy have similar acid type of the surface and are characterized by a high negative charge even at low values of pH of the dispersion medium. This may be determined by the presence of phosphate groups, teichoic acids being their carriers in Gram-positive bacteria. Glucose addition to peptone or glycerophosphate leads to the change of the bacteria surface type, i.e., at low values of pH the cells have total positive charge, that allows to suppose availability of protein nature components in the surface layers of the cell wall. Bacteria grown in these conditions are also characterized by the higher degree of adhesion to inert carrier (glass).

Bacillus subtilis↗

PubMed1991

[Myoglobinuria due to enzyme abnormalities in glycolytic pathway--especially lactate dehydrogenase M subunit deficiency].

Glycolysis is an important energy productive system. Enzyme abnormalities the in glycolytic pathway, which cause myoglobinuria, are deficiencies of phosphofructokinase, phosphoglycerate kinase, phosphoglycerate mutase, and lactate dehydrogenase (LDH). Common symptoms of these enzyme abnormalities are muscle cramp, muscle pain, and rhabdomyolysis after strenuous exercise. Acute renal failure owing to myoglobinuria is the most noteworthy symptom. In daily life, symptoms are rarely observed and prognosis is usually good. Correct and fast diagnosis of such latent symptomatic disorders is important to prevent a turn for the worse of these symptoms. LDH M subunit deficiency was first discovered by urinary discoloration and a discrepancy of laboratory data. Since then, only four cases have been reported in the Japanese population. The response to ischemic forearm work is characteristic (an increase of venous lactate concentration after ischemic work is not observed and a marked increase of venous pyruvate is found). The increase of pyruvate concentration is specific in LDH-M subunit deficiency, and is not observed in other abnormalities of the glycolytic pathway. Glycolysis was markedly retarded in the patient's muscle in the glyceraldehyde 3-phosphate dehydrogenase (GA3PD) step, possibly due to the impaired reoxidation of NADH produced by GA3PD activity. Then, the excess NADH is reoxidized by alpha-glycerophosphate dehydrogenase and triose phosphates are drained to alpha-glycerophosphate and glycerol. Therefore ATP production is significantly impaired and muscle tissue is damaged. A genetical study revealed a deletion of 20 base-pairs in exon 6 in LDH-M subunit deficiency. This mutation results in a frame-shift translation and premature termination.

Adolescent↗

PubMed1991

[Cytochemical characteristics of lymphocytes in peptic ulcer under various states of acid secreting function of the stomach].

Metabolic activity of lymphocytes was evaluated in ulcer patients with different acid-production function. Measurement of succinic dehydrogenase and alpha-glycerophosphate dehydrogenase of the mitochondria showed that acid production activity brought about variations in the activity of alpha-glycerophosphate dehydrogenase suggesting activation of a reserve path of the cell energy supply. The data obtained on the strain of the local defence and repair processes in gastric mucosa of patients with ulcer running with hyperacidity especially upon cicatrization of the lesion, provide a basis for the disease outcome.

Adult↗

PubMedDate not supplied

[Effect of removal and stimulation of the superior cervical sympathetic ganglia on the activity of oxidation-reduction enzymes in the neurosecretory cells of the anterior hypothalamus in rabbits].

Experiments were conducted on rabbits. A study was made of the activity of the redox enzymes--glucose-6-phosphate dehydrogenase (G-6-PDH), lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), malate dehydrogenase (MDH), NAD-and NADP-diaphorases, cytochromeoxidase (CCO), alpha-glycerophosphate dehydrogenase (alpha-GPDH) in the supraoptic and paraventricular nuclei of the hypothalamus and the posterior lobe of the hypophysis under conditions of stimulation and removal of the superior cervical sympathetic ganglia. There was revealed a correlation between the activity of the tissue respiration enzymes (SDH, MDH, NAD- and NADP-diaphorase, CCO) and the functional condition of the hypothalamo-neurohypophysial neurosecretory system. However, the enzymes of the pentose-phosphate (G-6-PDH) and glycerophosphate shunt (alpha-GPDH) and also of the anaerobic way of oxidation (LDH) reacted nonspecifically on the induced effects.

Animals↗

PubMedDate not supplied

Hypoxic right ventricular cardiomyopathy. A morphological and pathogenetic study on the myocardial atrophy and fatty infiltration.

The autopsy report of an asymptomatic, non familial cardiomyopathy with widespread fatty infiltration of the right ventricular wall in two alcoholic subjects, who were also heavy smokers and suffering from a serious laryngeal obstruction, led the Authors to investigate, on the basis of a thorough review of the literature, the possibility that hypoxia, alcoholism and smoke could have caused the development of the cardiac lesion. The presence of myocardial fatty infiltration is explained, under conditions of high-flow hypoxia, by the reduced fatty acid oxidation. The higher tissue levels of fatty acyl-CoA, fatty acyl-carnitine and alpha-glycerophosphate thereby lead to the increased conversion of the FFA into tissue lipids. Under hypoxic conditions there is also an increased polyols synthesis. The reduced conversion of dyacylglycerol into phosphatidic acid causes its tissutal increase and the interaction with fatty acyl-CoA to produce triacylglycerol and CoASH. In alcoholic patients reduced oxidation and increased FFA synthesis is sustained by the altered mitochondrial respiratory control and excess of acetate, with the consequent increase in acetyl-CoA, fatty acyl-CoA and alpha-glycerophosphate concentration. In addition, fatty acid ethyl esters normally absent in the myocardium are formed. The fact that, in hypoxic or alcoholic subjects with cardiomyopathy, an impaired myocardial contractility has been noted as the most relevant haemodynamic factor may be explained by both the reduced energy production following the decrease in aerobic glycolysis and FFA oxidation, and specific genetic changes that lead to both the production of a myosin with lower Ca2 + ATPase activity and a reduced protein (and therefore myofibrillar) synthesis. This fact can result in a severe atrophy of the cardiac myocytes. The lower their contractile activity, the more evident the process of atrophy. The lesion principally affects the right ventricle for both metabolic and anatomical reasons. It has been shown how, under normal conditions, the RV metabolism is suited to a relatively reduced O2 supply situation, with a high lactate dehydrogenase and alpha-hydroxybutiratedehydrogenase activity. It is more likely to be affected therefore whenever there is a chronic state of high-flow hypoxia. While alpha-HBDH allows the RV extensive utilization of ketone bodies as an energy source, its notable increase under hypoxic conditions further increases the synthesis of fatty acids and therefore fatty infiltration of the myocardium. The relatively lower capacity for oxygen extraction and lower tissue perfusion of the RV compared with the left ventricle make an adequate oxygen supply in the case of increased O2 demand even more difficult.(ABSTRACT TRUNCATED AT 400 WORDS)

Adipose Tissue↗

PubMed1978

A novel phosphodiesterase from Aspergillus niger and its application to the study of membrane-derived oligosaccharides and other glycerol-containing biopolymers.

A novel phosphodiesterase has been found in commercially available extracts of Aspergillus niger and has been partially purified by fractionation with acetone and chromatography on carboxymethylcellulose. The enzyme attacks glycerophosphodiester bonds with the liberation of free glycerol only. The synthetic substrate glucose 6-phospho-sn-1'(3')-glycerol is hydrolyzed with production of equivalent amounts of free glycerol and glucose 6-phosphate. Similarly, the enzymic hydrolysis of sn-glycero-3-phosphocholine liberates glycerol and phosphocholine. The hydrophilic head groups of membrane phospholipids of Escherichia coli are continuously transferred to a closely related family of oligosaccharides ("membrane-derived oligosaccharides") containing glucose as the sole sugar (van Golde, L. M. G., Schulman, H., and Kennedy, E. P. (1973) Proc. Natl. Acad. Sci. U. S. A. 70, 1368--1372). Oligosaccharide A-2 contains sn-1-glycerophosphate residues (derived from phosphatidylglycerol) in phosphodiester linkage. Treatment of this oligosaccharide with the phosphodiesterase led to the liberation of nearly all of the glycerol as free glycerol. Subsequent partial acid hydrolysis of the enzyme-treated oligosaccharide led to the recovery of glucose 6-phosphate in almost quantitative yield. The sn-1-glycerophosphate residues are therefore linked to position 6 of glucose units of the oligosaccharide. The activity of the enzyme is not restricted to glycerophosphodiesterases since it will hydrolyze phosphodiesters containing other polyols such as the synthetically prepared glucose 6-phospho-DL-1'(2'-hydroxy-3'-ethoxy)propane.

Aspergillus niger↗

PubMed1990

[Apatite-collagen complex. Preparation of a new apatite-collagen complex].

A new apatite-collagen complex was prepared in calcium beta-glycerophosphate solutions at pH 8.50. For this preparation, reconstituted type I collagen was cross-linked with phosvitin in the presence of alkaline phosphatase by use of a cross-linking agent of dimethyl suberimidate. After two weeks of immersion in daily-renewed solution of calcium beta-glycerophosphate, the complex contained apatite approximately two times the modified collagen in weight. When viewed in a scanning electron microscope, needle-like crystals precipitated densely on the collagen fibrils. However, in some portion of the complex, dot-like precipitate was observed as well. X-ray diffraction and IR analyses of the complex suggested that the apatite precipitated on the collagen fibrils was very similar to bone mineral in two aspects, crystallinity and carbonate content.

Apatites↗