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PubMed1993

Concentrations of nucleotides, nucleosides, purine bases and urate in cerebrospinal fluid of children with meningitis.

The release of agents mediating inflammation in meningitis may bring about neuronal hypoxia, under which circumstances ATP concentrations decrease and its degradation products increase and are released into the cerebrospinal fluid. In this study of alterations in neuronal energy metabolism in meningitis, AMP, IMP, inosine, adenosine, guanosine, adenine, guanine, hypoxanthine, xanthine and urate were determined by high performance liquid chromatography in the cerebrospinal fluid of 54 children aged between 1 month and 13 years suffering from meningitis (25 viral, 24 bacterial and 5 tuberculous cases) and 63 controls. Compared to the controls, patients with viral meningitis exhibited high concentrations of IMP, adenosine, guanosine, adenine, guanine and xanthine; patients with bacterial meningitis exhibited high concentrations of IMP, inosine, guanosine, adenosine, hypoxanthine, xanthine and urate; and patients with tuberculous meningitis exhibited high concentrations of AMP, guanosine, xanthine and urate. Viral and bacterial cases did not differ significantly for any of the metabolites studied. AMP and urate concentrations were significantly higher in patients with tuberculous cases compared with viral or bacterial meningitis cases.

Adenosine Monophosphate↗
PubMed2001

Concentrations of nucleotides, nucleosides, purine bases, oxypurines, uric acid, and neuron-specific enolase in the cerebrospinal fluid of children with sepsis.

To determine the effects of sepsis on cerebral energy metabolism, the cerebrospinal fluid adenosine monophosphate, inosine monophosphate, inosine, adenosine, guanosine, hypoxanthine, xanthine, and urate concentrations were determined by high-performance liquid chromatography and the neuron-specific enolase levels by means of an enzyme immunoassay method in 32 children with sepsis, without meningitis, aged between 2 months and 13 years, and in 160 age-matched controls. The septic group had significantly higher cerebrospinal fluid concentrations of inosine, adenosine, xanthine, and urate than controls. These results suggest that sepsis could provoke some degree of neuronal hypoxia and significant alterations of cerebral energy metabolism homeostasis.

Adolescent↗
PubMedDate not supplied

Nucleotide, nucleoside and purine base concentrations in human placentae.

Nucleotides, nucleosides and purine bases in trichloroacetic acid extracts of freeze clamped samples of human placenta have been measured by high-pressure liquid chromatography. The concentrations of the nucleotides concerned with energy transduction, ATP, ADP and AMP, and especially the energy charge, are stable over periods of ischaemia of 30 min. Concentrations of 14 nucleotides, including UDPAG, GDP Man, UDP and CTP, have now been defined. In addition, the concentrations of hypoxanthine, xanthine, uridine, adenine and inosine are indicated. Concentrations of the vasodilator adenosine are similar to the apparent Michaelis constants of its main metabolizing enzymes adenosine kinase and adenosine deaminase. The availability of 'normal' values of adenine nucleotide concentrations in human placenta should permit the detection of 'placental insufficiency' of energy supply, if this condition exists.

Cesarean Section↗
PubMed1979

Use of automated chromatography on the amino acid analyzer with lithium citrate buffers to separate nucleic acid bases, nucleosides, nucleotides and their precursors.

A simple and sensitive method has been developed to separate nucleic acid bases, nucleosides, nucleotides and their precursors by automated chromatography using the amino acid analyzer with lithium citrate buffers. The method is sensitive to a concentration of 5 nmol, linear in the range of 5--100 nmol, and resolves almost all the bases, nucleosides, nucleotides and their precursors of physiologic importance.

Autoanalysis↗
PubMedDate not supplied

Effect of a nucleoside-nucleotide mixture on protein metabolism in rats after seventy percent hepatectomy.

The effect of a nucleoside-based mixture (OG-VI) on protein synthesis in partially hepatectomized rats was evaluated. Twenty male Wistar rats weighing about 200 g underwent 70% hepatectomy by Higgins and Anderson's method, and then were subjected to total parenteral nutrition (TPN) for 3 days: the control group received only standard TPN (S-TPN group) and the experiment group received standard TPN supplemented with a nucleoside-nucleotide mixture (13% W/V, OG-VI) (OG-TPN group). The rates of whole body protein turnover and protein synthesis and breakdown were determined for 3 days after hepatectomy by the method of Picou and Taylor-Roberts with [15N] glycine. The nitrogen balances on days 1 and 3 and the cumulative nitrogen balance were significantly higher in the OG-TPN group than in the control group. Moreover, the rates of protein turnover and protein synthesis were significantly higher in the OG-TPN group than in the control group. Thus, increase in protein synthesis, rather than change in protein breakdown, was concluded to be responsible for the better nitrogen balance of the OG-TPN group. These findings suggest that the nucleoside-nucleotide mixture was used for the syntheses of RNA and DNA through the salvage pathway in regenerating liver and that this resulted in enhanced protein turnover with simultaneous increase in protein synthesis.

Animals↗
PubMed1993

Reversed-phase and ion-pair separations of nucleotides, nucleosides and nucleobases: analysis of biological samples in health and disease.

Methods for the assay of nucleotides, nucleosides and nucleobases in biological samples in health and disease are reviewed, with emphasis on reversed-phase and ion-pair reversed-phase techniques for their determination. Modes of extraction from biological samples are discussed with respect of the determination of in vivo concentrations. Advantages and limitations of ion-pair reversed-phase chromatography are discussed with examples from biochemistry and clinical chemistry. The capacity of the high-performance capillary electrophoresis is compared with that of ion-pair reversed-phase chromatography.

Animals↗
PubMed1995

A dietary nucleoside-nucleotide mixture inhibits endotoxin-induced bacterial translocation in mice fed protein-free diet.

Nucleosides and nucleotides are important substrates utilized by the intestinal mucosa. To determine the relative effect of dietary nucleosides and nucleotides on the gut, we investigated the effects of these compounds on endotoxin-induced bacterial translocation, cecal bacterial populations and ileal histology in protein-malnourished mice. There was an inhibition of gram-negative enteric bacteria in the mesenteric lymph node and spleen of the surviving mice fed the protein-free diet supplemented with a nucleoside-nucleotide mixture compared with the nonsupplemented group. Histologically, the damage to the gut mucosal barrier was more pronounced in the nonsupplemented group than in the nucleoside-nucleotide supplemented group. However, the cecal bacterial populations in the groups were not different. The villous height, crypt depth and total wall thickness were more developed in the supplemented group compared with the nonsupplemented group, indicating that the nucleoside-nucleotide mixture blocked bacterial translocation by preventing endotoxin-induced mucosal or epithelial damage. These results suggest that the nucleoside-nucleotide mixture could be used to inhibit or reduce the incidence of bacterial translocation, decrease intestinal injury and improve survival in a lethal model of protein deficiency and endotoxemia.

Animals↗
PubMed1996

A nucleoside-nucleotide mixture and its components increase lymphoproliferative and delayed hypersensitivity responses in mice.

We studied the effect of individual components of a nucleoside-nucleotide mixture on T cell-mediated immunity in BALB/c and DBA/2 mice. Mice were fed for 4 wk a nucleotide-free 20% casein diet (control) or this diet supplemented with 3 mol/kg of one of the following; inosine, guanosine monophosphate, uridine, cytidine, thymidine, or a mixture of these. In both strains of mice, popliteal lymph node immunoproliferative response to alloantigeneic (C57BL/6 splenic cells) challenge in mice fed the mixture and guanosine monophosphate was greater (P < 0.05) than in the mice fed the control diet. BALB/c mice fed inosine, uridine, and thymidine also showed greater (P < 0.05) responses compared with control fed mice. In the sheep red blood cells challenge assay, foot pad weight-gain in both strains of mice fed the mixture and the individual components supplemented diets was greater (P < 0.05) than in those fed the control diet. In both strains of mice, interleukin-2 secretion by popliteal lymph node lymphocytes in mice fed the mixture and thymidine was higher (P < 0.05) compared with the rest of the groups except BALB/c mice fed cytidine. Significantly higher than control secretions of interferon-gamma were observed only in BALB/c mice fed inosine, thymidine and the mixture. We conclude that mice fed the nucleoside-nucleotide mixture and the individual components of the mixture had greater responses in the T cell-mediated immune functions studied and that the responses in mice fed the mixture were not different from those in mice fed the individual components.

Animals↗
PubMed1997

The effect of a nucleotide-nucleoside solution on hepatic regeneration in rats after partial hepatectomy and in primary monolayer culture of hepatocytes.

Purine and pyrimidine metabolism is a key process after hepatic surgery. To evaluate the effect of purine and pyrimidine supplementation on hepatic regeneration, the following clinical in vitro and in vivo experiments were performed. Changes in blood nucleotides, nucleosides and nucleobase were analyzed by high performance liquid chromatography in patients and rats after partial hepatectomy. The effect of supplementation of nucleotide-nucleoside solution (OG-VI) or their components on nucleic acids syntheses in primary monolayer cultured hepatocytes and preoperative intraperitoneal injection of OG-VI on hepatic regeneration in the partially hepatectomized rats were evaluated. Blood purine and pyrimidine levels in patients change after hepatectomy and their changes indicate increased salvage synthesis of purine and pyrimidine in the regenerating liver. Addition of appropriate amounts of inosine, GMP, uridine, or thymidine, the substrates for salvage purine and pyrimidine syntheses, to primary cultures of hepatocytes enhanced both DNA and RNA syntheses by the salvage and de novo pathways. The OG-VI mixture also enhanced the syntheses of DNA and RNA. Preoperative administration of OG-VI to partially-hepatectomized rats enhances hepatic DNA synthesis in a way similar to the in vitro study.

Adenosine Triphosphate↗
PubMed1988

Extraction and measurement of myocardial nucleotides, nucleosides, and purine bases by high-performance liquid chromatography.

Nucleotides, nucleosides, and purine bases were extracted from human endomyocardial biopsies, freeze-clamped rat hearts, and porcine coronary sinus plasma. Perchloric acid extracts were neutralized with Freon-trioctylamine and analyzed at 250 nm by reverse-phase ion-pairing high-performance liquid chromatography. To achieve the sensitivity necessary for analyzing small (1-3 mg wet wt) tissue samples, a small-bore, 2.1-mm-internal-diameter, C18, 5-micron reverse-phase column and a flow rate of 0.2 ml/min were used. All of the myocardial nucleotides and AMP degradation products were resolved in a total separation time of 27 min with 30 mM KH2PO4, 7.5 mM tetrabutylammonium phosphate buffers, and binary pH and acetonitrile gradients.

Animals↗
PubMed1996

Nucleoside-nucleotide free diet protects rat colonic mucosa from damage induced by trinitrobenzene sulphonic acid.

BACKGROUND: Growing evidence suggests that intestinal recovery from injury induced by radiation, endotoxin, and protein deficiency is improved by the ingestion of nucleosides and nucleotides. AIM: This study examined the effect of dietary nucleosides and nucleotides supplementation on trinitrobenzene sulphonic acid induced colonic damage in experimental colitis. METHODS: Sprague-Dawley rats were randomised into two groups and fed nucleic acid free 20% casein diet (control) or this diet supplemented with 0.5% nucleoside-nucleotide mixture for four weeks. On the second week, colonic inflammation was induced in rats by intracolonic administration of 0.25 ml of 50% ethanol containing 25 mg of trinitrobenzene sulphonic acid. Additionally, other sets of rats were treated with 0.25 ml of 50% ethanol, 25 mg of trinitrobenzene sulphonic acid in 0.25 ml saline, or 0.25 ml of 0.9% saline. RESULTS: After two weeks, colon weight, macroscopic and microscopic damage scores, were significantly greater (p < 0.05) in the nucleoside-nucleotide supplemented group compared with the non-supplemented control groups. The same variables seen in the trinitrobenzene sulphonic acid-ethanol group fed nucleoside-nucleotide free diet were greater (p < 0.05) than in the rest of the groups fed nucleoside-nucleotide free diet and treated with ethanol, trinitrobenzene sulphonic acid in saline, or saline. Histologically, segmental ulceration and inflammation associated with significantly increased infiltration of polymorphonuclear leucocytes, macrophages, lymphocytes, fibroblasts were observed in the supplemented group compared with the controls. In the nucleoside-nucleotide supplemented group the epithelial damage, mucosal erosion, oedema, and coagulative necrosis of the muscularis propria was more extensive in comparison to the non-supplemented control groups. CONCLUSIONS: This study suggests that dietary nucleosides and nucleotides may aggravate colonic damage and inflammation in chemically induced experimental colitis in rats; and that nucleoside-nucleotide free diet combined with other pharmacological agents may offer a better response.

Animals↗
PubMed1994

[Effects of a nucleotide-nucleoside mixture on ischemic muscle metabolism in patients with stage II peripheral arterial occlusive disease. MR spectroscopic and biochemical analytic results].

In order to assess the acute metabolic effects of an intra-arterial infusion of nucleotide-nucleoside-mixture (NNM), 31P-mr-spectroscopy at the site of m. gastrocnemius and metabolite determinations from blood of the femoral artery and vein were carried out in 10 patients with PAOD stage II during ergometric calf exercise to the claudication pain limit. The spectroscopic measurements revealed a greater exercise-induced fall of PCr and a higher increase of Pi in calf muscles during supply of NNM compared with control ergometry. Post-exercise recovery of PCr was distinctly delayed during infusion of NNM. The anaerobic production of energy, however, was sufficient to maintain the ATP concentration to the same extent as under control ergometry. On the other hand, intramuscular lactate acidosis developed to a lower degree with NNM infusion than without NNM. A reduced muscular release of lactate, pyruvate, ammonia and alanine followed from the evaluation of the arteriovenous balance of these metabolites in the femoral vessels indicating a favourable global metabolic effect of NNM infusion in the extremity. The apparent contradiction in the spectroscopic and analytic-biochemical findings can be explained by local blood shunts induced by maximum vasodilation. Noninvasive mr-spectroscopy allows to detect directly and continuously the metabolic impact of ischemia in the calf muscles afflicted by arterial occlusion, whereas the metabolite concentrations in femoral blood are altered by afflux from non-ischemic areas. The known clinical benefit of frequently repeated intra-arterial infusions of NNM is thought to be due to an expansion of collateral circulation and to a favourable influence on endothelial functions.

Adenosine Triphosphate↗
PubMed1979

Changes in free nucleotides, nucleosides and bases during preparation of pre-cooked dehdyrated minced meats.

Changes in free nucleotides, nucleosides and bases during preparation of pre-cooked dehydrated minced meats from goat and sheep are reported. Major changes took place during cooking stage only; the changes during dehydration were relatively minor. Addition of EDTA at 500 ppm level significantly reduced the rate of dephosphorylation during curing process. Concentration of free nucleotides and related compounds were very low in commercially prepared accelerated freeze dried meat chunks.

Animals↗
PubMedDate not supplied

Cell growth arrest by nucleotides, nucleosides and bases as a tool for improved production of recombinant proteins.

Arresting cell growth and thus decreasing cell division potentially lessens the chance for genetic drift in the cell population; this would be of utmost importance for the consistent production of biopharmaceuticals during long periods. The drawback of the addition of well-known synchronizing agents, such as chemotherapeutics, is that they cause a disproportionate accumulation of cellular constituents, leading to cell death. The use of compounds that are naturally synthesized by the cell, as is the case of nucleotides, nucleosides, and bases (Nt/Ns/B), is shown in this work to be a promising tool. The addition of purines and pyrimidines was tested using a CHO cell line producing the secreted form of the human placental alkaline phosphatase enzyme (SEAP). From the chemical alternatives tested, AMP was the most promising compound for protein production improvement; it reduced cell growth and maintained the culture with high cell viability for long periods, while increasing SEAP specific productivity 3-fold. The use of CHO and BHK mammalian cells producing Factor VII and the use of a insect cell line (Sf9) showed that the effect of AMP addition seems to be independent of the r-protein and cell line. With the addition of AMP, accumulation of cells at the S phase was accompanied by an increase of the protein specific productivity. Addition of known synchronizing drugs (aphidicolin and doxorubicin) and application of environmental cell growth arrest strategies (depletion of nutrients and byproduct accumulation) showed also to effectively arrest CHO cell growth. A careful look onto cell cycle distribution in the different scenarios created, shows whether it is important to consider r-protein expression dependency upon cell cycle in process optimization and operation strategies.

Alkaline Phosphatase↗