Measurement of lactic acid in nanomolar amounts reliability of such a method as an index of glycolysis in pancreatic islets

• 1.1. The output of lactate is measured in isolated pancreatic islets.
• 2.2. The method for lactate assay is reproducible, with a coefficient of variation between duplicate measurements not exceeding 10%. The biological factors of variability are more important.
• 3.3. In islets exposed to glucose (16.7 mm), the output of lactate accounts for 45–50% of the uptake of glucose and its further conversion to triosephosphates.
• 4.4. Rapid, marked changes in lactate output occur when the glucose concentration of the incubation medium is modified.
• 5.5. The reliability of lactate output as an index of glycolysis in the islets is discussed.

     

The effect of d2o on glycolysis by rat hepatocytes

• 1.1. The effect of incorporating D₂O into the incubation medium on glycolysis and gluconeogenesis by hepatocytes from fasted rats was examined.
• 2.2. The substitution by heavy water, D₂O, at concentrations from 10 to 40%, stimulated glucose uptake, lactate production and CO₂ yields from glucose. At 10 mM glucose, 40% D₂O doubled glucose uptake, increased CO₂ production by 40%, and increased lactate production by 350%.
• 3.3. The stimulation of lactate production decreased at higher glucose concentrations, but was still substantial even at 80 mM glucose.
• 4.4. There was no effect on CO₂ production above glucose concentrations of 30 mM.
• 5.5. Ten percent D₂O showed little inhibition of lactate uptake, its oxidation and gluconeogenesis. At 40% D₂O the inhibition ranged from 10 to 20%.
• 6.6. No effect of D₂O on the rate of glucokinase or glucose-6-phosphatase was observed.
• 7.7. The concentration of fructose, 2,6-P was not affected by D₂O

     

Interaction of lipogenic substrates in porcine adipose tissue in vitro

• 1.1. Porcine adipose tissue was incubated with radiolabeled glucose, acetate or lactate. Saturation curves indicated that lactate > glucose > acetate in providing two-carbon units for fatty-acid synthesis.
• 2.2. Competition between individual substrates indicated that lactate was the best lipogenic substrate.
• 3.3. Incubation of all three substrates at concentrations observable in serum indicated that at 5.56mM, glucose was the preferred lipogenic substrate in the presence of 0.1 mM acetate and 1.0 mM lactate.
• 4.4. At elevated concentrations (18.52mM glucose, 1.0 mM acetate and 10.0 mM lactate), acetate and lactate were preferred to glucose as lipogenic substrates.

     

Aerobic glucose disposal in the oyster: An in vivo kinetic analysis

• 1.1. Aerobic glucose disposal in starved oysters exposed to 1 mM external glucose was 2.29 μg C/g wet wt/min.
• 2.2. It was hypothesized that the maximum disposal rate is limited by the maximum rate of transepithelial glucose transport.
• 3.3. The major recipients of glucose-carbon were glycogen and amino acids. 4. The rate of glucose-carbon disposal to these two pools was 0.80 and 0.42 μg C/g/min, respectively.
• 4.5. The internal energy state determines the pathways of glucose disposal.
• 5.6. Disposal of glucose-carbon in “glucose-primed” oysters is primarily into glycogen.
• 6.7. In fasted bivalves the disposal is primarily into amino acids and carboxylic acids.
• 7.8. The uptake of dissolved glucose has the potential of contributing significantly to growth under conditions where the external glucose concentration is kept artificially high.

     

Effect of diet and essential amino acids gavage on young rat amino acid metabolism enzymes

• 1.1. The effects of a high-fat, high-energy diet and essential plus semi-essential amino acid gavage on pup rats have been studied (60–65 animals).
• 2.2. The activities of alanine transaminase, adenylate deaminase, glutamine synthetase and serine dehydratase have been tested in liver and muscle.
• 3.3. Plasma was used for the estimation of proteins, urea, amino acids, glucose, lactate, 3-hydroxy-butyrate and acetoacetate.
• 4.4. Liver and muscle glutamine synthetase activities are increased by diet and gavage administered. Hepatic serine dehydratase is inhibited by a cafeteria diet but activated by amino acid gavage. Adenylate deaminase is inhibited by diet and gavage in the liver, but gavage does not affect this enzyme activity in muscle. Liver alanine transaminase is increased by the diet; in the muscle, cafeteria diet and amino acid gavage showed the highest values for this enzyme.
• 5.5. In the plasma, the increase in lactate produced by the diet is inhibited by the amino acids provided. Cafeteria-fed pups showed lower urea levels and higher 3-hydroxybutyrate concentrations in the plasma.
• 6.6. Intracellular glucose is diminished by cafeteria diet. In contrast, the blood cell amino acid concentration increases with diet and gavage supplied.

     

The peculiarities of nitrogen excretion in sturgeons (Acipenser ruthenus) (Pisces,Acipenseridae)—I. the influence of ration size

• 1.1. The nonfaecal nitrogenous excretion rate in starved sterlet fingerlings and fingerlings fed on different rations was investigated. The weight of the fish and temperature of the water was 43 g and 17.5°C, respectively.
• 2.2. In the nonfaecal excrements of starved sterlets the ammonia: urea ratio was substantially lower than in teleosts. This ratio was found to be 1.4:1.
• 3.3. In fed sterlets the urea excretion rate was higher than in starved ones but independent of ration size.
• 4.4. During the day the urea excretion rate in sterlets was constant.
• 5.5. The ammonia excretion rate accelerated 2 hr after feeding and reached its peak duration 6–11 hr after depending on the ration size.
• 6.6. Total ammonia output in the sterlet increased following the increase of ration size up to 8.4% of body wt. Further increases in ration size did not cause the corresponding elevation of ammonia excretion rate.

     

The effects of oxalate and glucose on lipogenesis by isolated hepatocytes from normal and biotin-deficient chicks (Gallus domesticus)

• 1.1. Isolated hepatocytes synthesize fatty acids and cholesterol from lactate and acetate with lactate being the more effective substrate.
• 2.2. Biotin deficiency decreased fatty add synthesis from both substrates but stimulated cholesterogenesis.
• 3.3. Exposure of intact hepatocytes to oxalate inhibited fatty acid and cholesterol synthesis from lactate, this effect was enhanced in biotin-deficient chicks. A similar effect was not observed when acetate was the substrate.
• 4.4. Synthesis of fatty acids from lactate and acetate was stimulated by glucose, biotin deficiency increased this response. Cholesterogenesis was reduced in control but not biotin-deficient chicks.

     

The effect of hypoxia on carbohydrate metabolism in flounder (Platichthys flesus L.)—I. Utilization of glycogen and accumulation of glycolytic end products in various tissues

• 1.1. Resting oxygen consumption at 10°C did not change from normoxia (150 mm Hg) down to an oxygen tension of 55 mm Hg for the flounder, Platichtys flesus.
• 2.2. Flounders exposed to hypoxia showed increased levels of blood glucose and lactate, dependent on the degree of hypoxia.
• 3.3. Due to hypoxia glycogen was depleted in the liver and swimming muscle but in the heart there was no significant change.
• 4.4. Liver glucose increased after 7 hr of hypoxia. Heart and muscle glucose did not change but the absolute glucose concentration in the heart was five times higher than in the muscle.
• 5.5. There is a transient accumulation of lactate in heart, liver and kidney after 7 hr of hypoxia while lactate accumulation in the swimming muscle is significant only after 21 hr of hypoxia.
• 6.6. Succinate only accumulated in the liver while alanine accumulated in muscle, heart and liver.

     

Fetal lung metabolism: Response to maternal fasting

• 1.1. Fetal lung metabolic response to maternal fasting late in gestation was investigated.
• 2.2. Maternal fasting 4 days before term was associated with low fetal plasma glucose and insulin levels but increased levels of fetal plasma glucagon, glycerol, lactate and fatty acids.
• 3.3. Fetuses from fasted mothers showed a significant decrease in body weight (30%), lung weight (30%) and lung glycogen (46%), but no change in lung protein, phospholipid or total lung DNA, suggesting that lung size is affected more than maturation.
• 4.4. Fetal lung slices incubated in vitro showed that lactate oxidation to CO₂ equalled that of glucose in control fetal lungs and was unaffected by maternal fasting, while glucose oxidation was depressed (23%).
• 5.5. Maternal fasting significantly decreased in vitro incorporation of [U-¹⁴C]-glucose, [U-¹⁴C]lactate and [1-¹⁴C]palmitate into lung phospholipids.
• 6.6. Fetal lungs from fasted mothers showed increased conversion of lactate to glucose, indicating gluconeogenic potential by fetal lung.
• 7.7. These studies show that plasma lactate serves as an important energy fuel and substrate for lipid synthesis for the fetal lung, and maternal fasting markedly alters fetal lung metabolism.

     

1H NMR study of metabolic alterations in the small intestine of rats infected with Hymenolepis diminuta

• 1.1. 1H NMR spectra of the duodenum, jejunum and ileum tissues of the small intestine of a rat showed metabolic gradients.
• 2.2. The concentrations of metabolites in these gut regions were altered by the presence of the tapeworm Hymenolepis diminuta.
• 3.3. In the infected duodenum there was significantly less glycogen, glucose and phosphocreatine/creatine, but significantly more lactate than in the corresponding controls.
• 4.4. Infected jejunum contained significantly less betaine but significantly more succinate, alanine and lactate.
• 5.5. Infected ileum had significantly less glycogen and taurine but significantly more alanine and lactate.

     

Regulation of glutamine catabolism in the perfused guinea-pig liver in relation to ureogenesis and gluconeogenesis

• 1.1. L-Glutamine conversion into ammonia, urea and glucose by the perfused liver of 48 hr starved guinea-pigs was concentration dependent attaining the maximal rate at 4 mM.
• 2.2. The activity of glutaminase I (EC 3.5.12), measured in isolated liver mitochondria was high enough to account for the observed rate of ammonia, urea and glucose formation by the perfused liver. Neither NH4C1 (5 mM) nor aminooxyacetate (0.5 mM) affected the rate of glutamine conversion into glutamate by isolated liver mitochondria.
• 3.3. Gluconeogenesis and ureogenesis from glutamine was inhibited by octanoate, Dt-3-hydroxybutyrate, aminooxyacetate, ethanol and p-hydroxyphenylpyruvate while ammonia formation was stimulated by aminooxyacetate. 2,4-Dinitrophenol stimulated the rate of the formation of all three metabolites from glutamine.
• 4.4. The major changes induced by aminooxyacetate, as determined in livers perfused with glutamine and stopped by freeze-clamping technique, consisted in a decrease in the content of ATP, aspartate and malate and in a slight increase in the content of glutamate.
• 5.5. Glutamine is an effective precursor of phosphoenolpyruvate in isolated liver mitochondria. Its formation was inhibited by octanoate and by DL-3-hydroxybutyrate.
• 6.6. The data are discussed in terms of regulation of glutamine catabolism in liver with emphasis on ureogenesis and gluconeogenesis.

     

Effect of adenosine on gluconeogenesis in the perfused liver of chicken and guinea-pig

• 1.1. Glucose formation from lactate by the perfused liver of 48 hr starved chickens was strongly inhibited by adenosine (Ado); the half-maximal inhibition was attained at 40 μM. This effect was paralleled by a four- to five-fold increase of ATP content as determined in freeze-clamped liver.
• 2.2. In chicken liver homogenate gluconeogenesis from precursors such as alanine, glutamate, glutamine and aspartate, which are not converted into glucose by the perfused chicken liver, proceeded at rates equal to or higher than that with lactate, being markedly inhibited by Ado.
• 3.3. In the perfused guinea-pig liver glucose synthesis with lactate, propionate, glycerol and fructose was also inhibited by Ado; however, when precursors such as pyruvate, glutamine and a mixture of lactate + pyruvate were supplied to the liver Ado did not inhibit gluconeogenesis.
• 4.4. Assay of adenine nucleotides in the perfused guinea-pig liver, stopped by freeze-clamping technique in a number of experimental variants, revealed no correlation between the rate of gluconeogenesis and the changes induced by Ado in the adenine nucleotide pool.
• 5.5. In the perfused liver of both chicken and guinea-pig Ado produced an increase of the lactate to pyruvate ratio and, in general, a diminution of the content of malate-aspartate shuttle intermediates.
• 6.6. The results are interpreted as suggesting that the inhibitory effect of Ado on hepatic gluconeogenesis is not necessarily mediated by the changes in the adenine nucleotide pool.

     

Increased muscle glucose uptake in response to chronic glyburide treatment is not related to changes in glucose transporter (GLUT4) protein

• 1.1. The mechanism of action of glyburide (a sulfonylurea) on muscle has been investigated by measuring glucose uptake and glucose transporter (GLUT4) protein levels after chronic glyburide treatment.
• 2.2. A dietary induced insulin resistant rat model (4 wk of high-fat, high-sucrose feeding) was given glyburide (2mg/kg/day) for 10 days and glucose uptake was measured in a perfused hindquarter preparation.
• 3.3. Protein levels of the GLUT4 glucose transporter were determined by Western analysis.
• 4.4. After 7 days of treatment, rats fed glyburide had lower blood glucose concentrations 2 hr (72 ± 5 vs 103 ± 12 mg/dl) and 24 hr (97 ± 7 vs 123 ± 7 mg/dl) after glyburide administration with no difference in serum insulin levels compared to vehicle treated animals.
• 5.5. Glucose uptake was approx doubled in basal state (0 insulin) in response to glyburide (2.8 + 0.4 vs 1.7 ± 0.2μ mol/g per hr).
• 6.6. Maximal insulin (100 nM) stimulated glucose uptake tended to be higher in the glyburide treated group, but did not reach statistical significance (8.0 ± 0.7 vs 7.0 ± 0.6 μmol/g per hr).
• 7.7. Western analysis revealed no significant effect of glyburide on the GLUT4 protein level in skeletal muscle.
• 8.8. These results suggest that glyburide alters glucose uptake through some mechanism other than alterations in the level of the GLUT4 glucose transporter protein.

     

The role of citrate derived from glucose in the acetylcholine synthesis in rat brain synaptosomes

• 1.1. Synaptosomes utilizing glucose or glucose plus malate produced citrate with rates of 2.4 and 7.8 nmol/hr/mg of protein, respectively.
• 2.2. (−)Hydroxycitrate increased citrate net synthesis 4 times and inhibited acetylcholine synthesis by 40%.
• 3.3. Oxygen and glucose consumption as well as lactate and CO₂ production were not changed by this inhibitor.
• 4.4. (−)Hydroxycitrate inhibited utilization of exogenous citrate in synaptosomes by 50%.

     

Glycolytic enzymes of fowl and turkey spermatozoa

• 1.1. It was confirmed that, under anaerobic conditions, fowl spermatozoa formed lactate from glucose thirteen times faster than turkey spermatozoa.
• 2.2. The profiles of glycolytic enzyme activities were similar for spermatozoa from both species; however fowl spermatozoal activities were generally 2- to 4-fold higher.
• 3.3. Exceptions were glycerophosphate mutase and lactate dehydrogenase activities which were respectively 9.5 and 41 times greater in fowl spermatozoa.
• 4.4. In both species, spermatozoal glyceraldehyde-3-phosphate dehydrogenase had the lowest activity of the glycolytic enzymes.

     

Serum chemistry profiles for Lechwe waterbucks (Kobus leche): Variations with age and sex

• 1.1. Over an 8-year period, 19 biochemical parameters have been determined at various ages in the blood serum of 92 clinically healthy Lechwe waterbucks (Kobus leche), 33 males and 59 females.
• 2.2. Significant differences have been noted with age. In neonates, the lowest values of total proteins, glucose, creatinine, urea, AST, ALT and iron have been noted; the highest ones have been seen for cholesterol, alkaline phosphatase, calcium and phosphorus.
• 3.3. With regard to sex, raised values of glucose, urea, alkaline phosphatase and ALT, and lowered values of cholesterol, have been noted in juvenile females compared with males of the same age.
• 4.4. In adult females, higher levels of urea and cholesterol and lower levels of glucose, triglycerides and natrium have been recorded compared with males.
• 5.5. With sex and age, no significant changes have been found in the levels of GGT, magnesium, chlorides and copper.
• 6.6. Out findings are discussed with those abstracted from the literature for related species.

     

Urea excretion rates and waste nitrogen concentrations during metamorphosis of Xenopus laevis Daudin

• 1.1. Serum urea, ammonia concentrations in the blood and excretion were measured in tadpoles of different stages and juveniles of Xenopus laevis.
• 2.2. The urea excretion rate was determined with the help of injected ¹⁴C-urea.
• 3.3. Urea concentrations are higher during metamorphic climax and at the end of metamorphosis than during prometamorphosis.
• 4.4. Blood ammonia levels remain rather constant throughout metamorphosis.
• 5.5. Coincidentally, the relative amount of urea in the blood increases.
• 6.6. The ¹⁴C-urea excretion rates slow down from very high values (48%/hr) at the beginning of prometamorphosis to low rates (5%/hr) in newly metamorphosed animals.
• 7.7. This means that during metamorphosis not only is the possibility of urea production established. but there is a capacity to retard and store urea to some extent.

     

Regulation of spermidine transport in l1210 cells

• 1.1. 1 mM 2-amino isobutyric add (AIB), glutamine or asparagine when preincubated for 3 hr with L1210 cells promoted a marked increase in the rate of spermidine uptake.
• 2.2. Cycloheximide also increased the transport rate and completely prevented the increase due to AIB.
• 3.3. Trifluoperazine and iso-H7 inhibited the uptake of spermidine, much less the uptake of AIB.
• 4.4. Adenosine promoted an increase in the uptake of AIB, a decrease in that of spermidine.
• 5.5. Hypotonic stress also increased the rate of spermidine transport. This modification was only partially prevented by cycloheximide.
• 6.6. Okadaic arid had no effect on this increase, whereas it prevented the increase of ODC activity.

     

2-Deoxyglucose transport by the frog sartorius: Effects of electrical stimulation and N-carbobenzoxy-glycyl-l-phenylalaninamide

• 1.1. Studies characterizing glucose transport in the frog sartorius were performed.
• 2.2. For nonstimulated and stimulated muscles, intracellular 2-deoxyglucose exceeded 2-deoxyglucose-6-phosphate at 15 min, showed little further increase, and was maintained below the extracellular concentration for 2 hr.
• 3.3. Accumulated 2-deoxyglucose-6-phosphate did not inhibit glucose transport.
• 4.4. Unlike in adipocytes, basal and stimulated 2-deoxyglucose transport showed no difference in sensitivity to N-carbobenzoxy-glycyl-l-phenylalaninamide.
• 5.5. Phenylarsine oxide blocked contraction-enhanced 2-deoxyglucose uptake.
• 6.6. These results suggest that the glucose transporter of the sartorius exhibits auto-regulation, and that basal transport is not regulated by the same process as in adipocytes.

     

Changes in mammary glucose and protein uptake in relation to milk synthesis during lactation in high- and low-yielding goats

• 1.1. Glucose and protein uptake were measured in both mammary glands of two low- and two high-yielding dairy goats during lactation.
• 2.2. Low-yielding goats tended to have higher arterial glucose concentrations, but approximately 40% lower arterio-venous differences (AV) and extraction rates (E) for glucose than high-yielding goats.
• 3.3. AV and E for glucose (but not protein) were linearly related to yields of both lactose, milk protein and fat.
• 4.4. Mammary uptake of glucose is determined primarily by mammary glucose metabolism, not glucose supply; lower intracellular glucose concentration in mammary cells of genetically superior animals thus explains the more efficient mammary uptake.