Iotrol, iodixanol, and 2-deoxy-D-glucose effects on neural tissue CO2 production.

In vivo and in vitro experiments have demonstrated that the myelographic agent metrizamide decreases neural tissue glucose metabolism whereas iohexol and iopamidol do not. This study compares the changes in slices of rat hippocampus CO2 production caused by the nonionic dimers iotrol and iodixanol with the effects of metrizamide and 2-deoxy-D-glucose. After 6-hr incubations, 70-mmol/l concentrations of iotrol and iodixanol increased CO2 production by 11 +/- 20% and 31 +/- 35%, respectively, as compared with the artificial CSF control medium. Metrizamide at 70 mmol/l and 2-deoxy-D-glucose at 35 mmol/l decreased CO2 production by 32 +/- 13% and 96 +/- 1%, respectively. The increases in CO2 production with iotrol and iodixanol appear to indicate that these molecules have some effect on cell metabolism. The mechanism for the increase in CO2 production could involve an effect on the glucose metabolic pathway or could be indirect via a mechanism that increases cell energy utilization. These in vitro effects have not been verified with in vivo experiments.

Effects of intravenous glucose loading on oxygen consumption, carbon dioxide production, and resting energy expenditure in infants with bronchopulmonary dysplasia.

To determine the effects of intravenous glucose loading on basal oxygen consumption, resting energy expenditure, and basal carbon dioxide production in infants with bronchopulmonary dysplasia who were still oxygen dependent, we administered intravenous glucose loads of 4 and 12 mg/kg-1/min-1 on 2 consecutive days, under identical experimental conditions, to six infants with bronchopulmonary dysplasia and six healthy control subjects. Infants were not fed for 9 hours before and during the 4- to 6-hour study periods; the intravenous glucose infusion, along with an amino acid mixture (2 gm.kg-1.24 hr-1), was started at the beginning of the fasting period. Oxygen consumption and carbon dioxide production and resting energy expenditure were measured by a flow-through indirect calorimetry technique under basal conditions. Infants with oxygen-dependent bronchopulmonary dysplasia had significantly higher basal oxygen consumption and resting energy expenditure than did control infants and significantly higher basal carbon dioxide production during the high glucose infusion. With glucose loading, infants with bronchopulmonary dysplasia had a significant rise in basal oxygen consumption (7.91 +/- 0.91 ml.kg-1.min-1 to 9.65 +/- 1.35 ml.kg-1.min-1, p less than 0.05), basal carbon dioxide production (5.93 +/- 0.72 ml.kg-1.min-1 to 7.10 +/- 1.04 ml.kg-1.min-1), and resting energy expenditure (53.8 +/- 5.75 kcal.kg-1.24 hr-1 to 65.3 +/- 7.0 kcal.kg-1.24 hr-1, all p values less than 0.05). Control infants had no significant changes with intravenous glucose loading. We conclude that intravenous glucose loading in infants with bronchopulmonary dysplasia resulted in a net increase in resting energy expenditure, which should be taken into account in assessing their energy intake during nutritional management. The risk of pulmonary stress caused by an increase in basal oxygen consumption and carbon dioxide production resulting from glucose load should also be considered.

Functional hepatocyte heterogeneity. Vascular 2-oxoglutarate is almost exclusively taken up by perivenous, glutamine-synthetase-containing hepatocytes.

1. In isolated perfused rat liver maximal rates of 2-[1-14C]oxoglutarate uptake were about 0.4 mumol.g-1 .min-1; half-maximal rates of 2-[14C]oxoglutarate uptake were observed with influent concentrations of about 100 microM. 2-[14C]Oxoglutarate uptake by the liver was not affected by the direction of perfusion, but was decreased by about 80-90% when Na+ in the perfusion fluid was substituted by choline+, suggesting a Na+-dependence of hepatic 2-oxoglutarate uptake. In the absence of added ammonia, [14C]oxoglutarate uptake by the liver was about twice the net oxoglutarate uptake, indicating a simultaneous release of unlabeled oxoglutarate from perfused rat liver. 2. 14C-Labeled metabolites derived from [1-14C]oxoglutarate and recovered in the effluent perfusate were 14CO2 and 14C-labeled glutamate and glutamine; they accounted for 85-100% of the radiolabel taken up by the liver. 14CO2 was the major product (more than 70%) from [1-14C]oxoglutarate taken up the liver, provided glutamine synthesis was either inhibited by methionine sulfoximine or the endogenous rate of glutamine production was below 40 nmol.g-1.min-1. 3. Stimulation of glutamine synthesis by ammonia did not affect [14C]oxoglutarate uptake by the liver, but considerably increased net hepatic oxoglutarate uptake, indicating a decreased release of unlabeled oxoglutarate from the liver. Stepwise stimulation of hepatic glutamine synthesis led to a gradual decrease of 14CO2 production and radiolabel was recovered increasingly as [14C]glutamine in the effluent. At high rates of glutamine formation (i.e. about 0.6 mumol.g-1.min-1), about 60% of the [1-14C]oxoglutarate taken up by the liver was recovered in the effluent as [14C]glutamine. 14CO2 and [14C]glutamine production from added [1-14C]oxoglutarate were dependent on the rate of hepatic glutamine synthesis but not on the direction of perfusion. Extrapolation of 14C incorporation into glutamine to maximal rates of hepatic glutamine synthesis yielded an about 100% utilization of the [14C]oxoglutarate taken up by the liver for glutamine synthesis. This was again true for both the antegrade and the retrograde perfusion directions. On the other hand, addition of ammonia did not affect 14CO2 production from labeled oxoglutarate, when glutamine synthetase was inhibited by methionine sulfoximine. 4. The data suggest that vascular oxoglutarate is almost exclusively taken up by the small perivenous hepatocyte population containing glutamine synthetase, i.e. a cell population comprising only 6-7% of all hepatocytes. Thus, the findings demonstrate the existence of a, to date, uniquely zonally distributed oxoglutarate transport system which is probably Na+-dependent in the plasma membrane.(ABSTRACT TRUNCATED AT 400 WORDS)

Whole body gas exchange: amino acid and lactate clearance as indicators of initial and early allograft viability in liver transplantation.

A method for the rapid assessment of liver allograft circulation and function after liver transplantation is described. In 12 patients undergoing liver transplantation continuous recording of whole body energy production was made on the basis of gas exchange measurements during the surgical procedure. Oxygen consumption decreased rapidly by 25% when the blood supply to the native liver was interrupted. After the anhepatic period, there was a sharp increase of oxygen consumption with successful reperfusion of the allograft. Carbon dioxide production fell by 14% and returned to preanhepatic values after successful declamping. In two cases with suboptimal reperfusion the return of gas exchange values was slow and incomplete. Results from the studies of whole body energy production were compared with biochemical measurements. No significant accumulation of amino acids occurred during the anhepatic period, but in the two patients with incomplete revascularization, clearance of amino acids, after the anhepatic phase, was impaired and plasma amino acids accumulated. The same pattern was found for plasma lactate levels. By the techniques described in this article, rapid and reliable assessment of initial and early graft function in hepatic transplantation is possible. This is of great value for the intraoperative and early postoperative assessment and planning of surgical and anesthesiologic strategies.

Metabolism of aspartate in Mycobacterium smegmatis.

Mycobacterium smegmatis grows best on L-asparagine as a sole nitrogen source; this was confirmed. [14C]Aspartate was taken up rapidly (46 nmol.mg dry cells-1.h-1 from 1 mM L-asparagine) and metabolised to CO2 as well as to amino acids synthesised through the aspartate pathway. Proportionately more radioactivity appeared in the amino acids in bacteria grown in medium containing low nitrogen. Activities of aspartokinase and homoserine dehydrogenase, the initial enzymes of the aspartate pathway, were carried by separate proteins. Aspartokinase was purified as three isoenzymes and represented up to 8% of the soluble protein of M. smegmatis. All three isoenzymes contained molecular mass subunits of 50 kDa and 11 kDa which showed no activity individually; full enzyme activity was recovered on pooling the subunits. Km values for aspartate were: aspartokinases I and III, 2.4 mM; aspartokinase II, 6.4 mM. Aspartokinase I was inhibited by threonine and homoserine and aspartokinase III by lysine, but aspartokinase II was not inhibited by any amino acids. Aspartokinase activity was repressed by methionine and lysine with a small residue of activity attributable to unrepressed aspartokinase I. Homoserine dehydrogenase activity was 96% inhibited by 2 mM threonine; isoleucine, cysteine and valine had lesser effects and in combination gave additive inhibition. Homoserine dehydrogenase was repressed by threonine and leucine. Only amino acids synthesised through the aspartate pathway were tested for inhibition and repression. Of these, only one, meso-diaminopimilate, had no discernable effect on either enzyme activity.

Proton translocation coupled to the oxidation of carbon monoxide to CO2 and H2 in Methanosarcina barkeri.

Cell suspensions of acetate-grown Methanosarcina barkeri mediate the conversion of CO and H2O to CO2 and H2. The reaction is coupled with the phosphorylation of ADP. Evidence is presented that CO oxidation by the cells is associated with the transient acidification of the suspension medium. Up to 2 mol vectorial protons were measured/mol CO oxidized when the transmembrane electrical gradient was kept low by the addition of valinomycin (20 nmol/mg protein) and KCl (200 mM) or of KSCN (50 mM). No transient acidification was observed in the presence of the protonophore tetrachlorosalicylanilide which stimulated rather than inhibited CO oxidation. Proton extrusion remained unaltered when the proton-translocating ATPase was specifically inhibited by dicyclohexylcarbodiimide. The latter finding indicates that proton translocation is associated with CO conversion to CO2 and H2 rather than with ATP hydrolysis in the cells. The data substantiate that the coupling of CO oxidation with ADP phosphorylation in M. barkeri occurs via a chemiosmotic mechanism.

Propionic acidemia--biochemical studies.

The first documented case of propionic acidemia in Israel is described. Diagnosis was based on three independent methods: analysis of urinary organic acids by gas chromatography/mass spectrometry, assay of propionyl CoA carboxylase activity and oxidation of 1-14C-propionate by cultured skin fibroblasts. The use of more than one method for confirmation of the diagnosis is considered to be of importance in providing an additional margin of safety in cases where genetic counseling and prenatal diagnosis in future pregnancies are indicated.

Hyper and hypodynamic models of sepsis in guinea pigs.

The acute metabolic response following experimentally induced sepsis can generally be classified as either hypodynamic ("low flow") or hyperdynamic ("high flow"). We have found that in conscious guinea pigs the bolus infusion of 10(10) live Escherichia coli bacteria can elicit either response, depending on the route of administration of the bacteria. Intravenous infusion results in the hypodynamic condition of septic shock in which oxygen consumption (VO2) is reduced to approximately 60% of the control level, plasma glucose is elevated 4 hr after infusion with a reversal to extreme hypoglycemia 12 hr after infusion, and body temperature is reduced by approximately 5 degrees C in 12 hr. In contrast, subcutaneous injection results in increased VO2, body temperature, and plasma glucose. In both models the concentration of cortisol, catecholamines and glucagon were elevated, but the responses were more pronounced in the hypodynamic model. In both cases, insulin concentration was decreased. These models of sepsis are useful because many aspects of response are comparable to man, they are simple to create, and they are consistent and reproducible.

Lipogenesis in liver and adipose tissue of the domestic cat (Felis domestica).

1. Production of fatty acids, CO2 and glycerol from glucose and acetate was determined in slices of liver and adipose tissue taken from mature cats. 2. Acetate was the predominant carbon source for de novo fatty acid synthesis in both liver and adipose tissue. 3. Fatty acid synthesis occurred at much greater rates in adipose tissue than in liver. 4. Relative lipogenic capacity (ratio of production of long-chain fatty acids and glycerol to oxidation of substrates) was 20 times greater in adipose tissue than in liver with both substrates.

Effect of dietary bulk on organ mass, fasting heat production and metabolism of the small and large intestines in sheep.

Groups of 10 lambs were fed either a control diet or a diet consisting of 10% polyethylene at isoenergetic levels for 30 d to study the effect of dietary bulk on visceral organ mass and metabolism of mucosa from the jejunum and proximal colon. Fasting heat production was measured using an additional six lambs. Weights of the large intestine, stomach complex and lungs were greater in lambs fed the polyethylene diet than control lambs. Liver and small intestine weights were not affected by diet. Oxygen consumption rates of jejunal mucosa were not affected by diet while a trend existed for oxygen consumption rates of mucosa from the proximal colon to be increased due to feeding polyethylene. Na+,K+ ATPase activity of mucosa from both jejunum and colon was increased due to feeding polyethylene. Fasting heat production, oxygen consumption or carbon dioxide production did not differ with diet. Dietary bulk affects growth and metabolism of visceral organs but metabolic modifications appear different among tissues.

Effect of thiouracil in modifying folate function in severe vitamin B12 deficiency.

The effects of thiouracil in correcting defects in folic acid function produced by B12 deficiency were studied. Addition of the thyroid inhibitor, thiouracil, to a low methionine diet containing B12, increased the oxidation of [2-14C]histidine to carbon dioxide, and increased liver folate levels. Addition of 10% pectin to the diet accentuated B12 deficiency as evidenced by a greatly decreased rate of histidine oxidation (0.19%) and an increased excretion of methylmalonic acid. Addition of thiouracil to the diet restored folate function as measured by increased histidine oxidation and increased liver folate levels similar to that produced by addition of methionine to a B12-deficient diet. Thiouracil decreased methylmalonate excretion, and increased hepatic levels of B12 in animals on both B12-deficient and -supplemented diets. Hepatic methionine synthase was increased by thiouracil, which may be the result of the elevated B12 levels. S-Adenosylmethionine and the enzyme methionine adenosyltransferase were also increased by thiouracil. Thus it is possible that the effect of thiouracil in increasing folate function consists both in the effect of thiouracil in decreasing levels of methylenetetrahydrofolate reductase, and also in its action in increasing S-adenosylmethionine which exerts a feedback inhibition of this enzyme.

Challenge of phagocyte metabolism by extracorporeal circulation and membrane contact. A biocompatibility test.

In the present study, direct data on phagocyte metabolism during hemodialysis are collected by the determination of 14CO2-production from labelled glucose on whole blood samples. Fifteen minutes after the start of cuprophan dialysis, 14CO2 production per 10(3) phagocytes increased more than four-fold (from 77.6 +/- 30.2 to 315.0 +/- 69.6 DPM/10(3) phagocytes; P less than 0.01). Such a change was absent during dialysis with membranes containing polysulphone, AN69S or reused cuprophan. In vitro contact of normal blood with cuprophan membranes revealed an increase of 14CO2-production by a maximum of 9.0 +/- 3.4 X 10(3) D.P.M. at 15 min. (P less than 0.05). Such an increase was absent after contact with polysulphone. The measurement of 14CO2 production during glucose metabolism, by phagocytes present in micro-amounts of whole blood, is a valuable test for membrane biocompatibility. Cuprophan dialysis induces a challenge of phagocytic activity that is absent for dialysers containing other membranes.

Real importance of alanine in renal metabolism: in vitro studies in rat and dog.

In vitro studies were performed on cortical renal tubules to clarify possible differences between dog and rat with regard to alanine production and to define more precisely the role of alanine on ammonia and glucose production by the kidney. It was established that glutamate-pyruvate transaminase has an activity that is seven times lower in the rat than in the dog kidney. At the same time, alanine production from lactate, pyruvate, and glutamate is three times lower in the rat than in the dog kidney. The enzymatic reaction could be completely inhibited in a competitive fashion with aminooxyacetate. O2 consumption and CO2 production by the renal tubules were lower than that observed with glutamine. CO2 production in the rat was lowest. Production of ammonia and glucose by the kidney from alanine during acidosis averaged less than 20% of that produced with L-glutamine. Furthermore, during metabolic acidosis the production of ammonia and glucose from alanine was not augmented and failed to be influenced by increasing the concentration of alanine in the incubation medium.

CO2 production of the chick embryo during the first day of post-laying development.

The carbon dioxide production of the chick embryo cultured in vitro has been determined during the first 24 h of post-laying development using a non-invasive conductometric microtechnique. The mean CO2 production of the whole blastoderm (1) increased from 16 nmol/h at laying to 231 nmol/h at early neurulation, (2) became dependent on exogenous glucose and (3) was closely linked to mechanical tension generated in the blastoderm (loosening from vitelline membrane resulted in a decrease of 56%). In our experimental conditions, no significant influence of carbonic anhydrase on the CO2 production has been detected. The value of the respiratory exchange ratio varied from about 3 at pregastrular stages to 1 at neurula stage and CO2 was produced transiently in presence of antimycin A. Such results indicate that the source of CO2 is not exclusively mitochondrial and that the relative proportions of mitochondrial and non-mitochondrial CO2 productions might vary significantly throughout the early development. Our findings confirm that the metabolism of the chick embryo becomes more and more oxidative from laying onwards and suggest that the modifications of metabolism observed during the studied period of development could be associated with functional differentiation.