Search PubMed⌕ Search

Biomedical subjects

J L Robinson

Publications and source records attributed to J L Robinson.

At least 109 records · Page 6Linked to original sources

Hepatic metabolites and amino acid levels during adaptation of rats to a high protein, carbohydrate-free diet.

Changes in hepatic levels of lactate, pyruvate, phosphoenolpyruvate (PEP), alpha-ketoglutarate, malate, oxaloacetate, ketone bodies, alanine, serine, glycine, aspartate, glutamate, glutamine, valine, urea, adenine nucleotides and inorganic phosphate were examined in rats consuming a high protein, carbohydrate-free diet for up to 40 days. While some components showed transient changes, others (pyruvate, malate, oxaloacetate, PEP, ketone bodies, alanine, glycine, glutamine, valine, urea, adenine nucleotides and inorganic phosphate) were permanently altered. The cytoplasmic and mitochondrial redox states were only transiently affected and by day 24 were not different from control values. In contrast, the cytoplasmic phosphorylation state was affected from day 1 on; this suggests a role for the latter in permanently reorienting metabolism toward gluconeogenesis and ureogenesis.

Adaptation, Physiological↗

Circadian variation of liver metabolites and amino acids in rats adapted to a high protein, carbohydrate-free diet.

Diurnal variation in hepatic levels of lactate, pyruvate, phosphoenolpyruvate (PEP), alpha-ketoglutarate, malate, oxaloacetate, ketone bodies, alanine, serine, glycine, aspartate, glutamate, glutamine, valine, urea, adenine nucleotides and inorganic phosphate were studied in rats adapted to a high protein, carbohydrate-free diet for 24 days. Most circadian rhythms differed in relation to controls (10% protein diet); many merely had different amplitudes, some were inverted, and some exhibited drastically altered patterns. Cytoplasmic redox state exhibited nearly similar variations and phosphorylation state differed primarily in amplitude whereas mitochondrial redox state was highly depressed in the absorptive phase. The metabolic regulation implied by the results is discussed in relation to both circadian variations of plasma insulin and glucagon concentrations, and pyruvate kinase and phosphoenolpyruvate carboxykinase activities previously reported.

Adenine Nucleotides↗

Quantitative gait evaluation in the clinic.

The purpose of this paper is to present a clinically feasible method to provide objective, quantitative information regarding the quality of a patient's gait. The technique used is simple and economical and requires minimal equipment. The four basic temporal and distance factors are 1) stride length, 2) step length, 3) cadence, and 4) velocity. The technique requires the following equipment: 1) grid pattern, 2) stop watch, 3) portable tape recorder, 4) tape measure, 5) marking pen, and 6) masking tape. Case examples are provided to demonstrate the use of the data obtained.

Adult↗

Effect of amino acid concentration on orotic acid production by bovine mammary tissue.

Investigations were to determine if bovine mammary tissue could synthesize orotate and to assess the influence of amino acids on production of orotate. Mammary tissue from 6 Holstein cows was obtained immediately after slaughter. Tissue slices were incubated in either a modified Krebs-Ringer bicarbonate buffer, the same medium containing a physiological concentration of amino acids, or a third medium containing a fourfold concentration of the amino acid mixture. In all experiments, tissue slices were preincubated in the physiological amino acid medium for 5, 10, and 15 min (30 total min) with fresh medium changes after each incubation with one of the three treatment media for three consecutive 20-min periods. Tissue slices incubated in the physiological amino acid medium for a total of 90 min synthesized 28.5 microgram orotate/g wet tissue after correction for the initial tissue contribution. Orotate secreted by the tissue slices increased as concentration of amino acids increased. Lactose released into the medium was not affected by amino acid concentration. Bovine mammary tissue can synthesize substantial amounts of orotate, and tissue synthesis of orotate but not lactose increased as amino acid concentration increased.

Amino Acids↗

Development of gluconeogenic enzymes in the liver of fasting or suckling newborn pigs.

Gluconeogenic enzymes and substrates were measured in the livers of fasted and suckled newborn pigs in the first 48 h postpartum. The activities at birth of glucose-6-phosphatase, fructose-1,6-diphosphatase, pyruvate carboxylase and phosphoenolpyruvate carboxykinase were, respectively, 70%, 45%, 117% and 35% of adult values. At birth, cytosolic phosphoenolpyruvate carboxykinase represented 35% of total activity, a similar distribution to that in the adult. In suckled piglets, all activities were greater at 24 and 48 h that at birth. In starved piglets, the increases were greater in all cases; the increase in cytosolic phosphoenolpyruvate carboxykinase was much more pronounced than for that for the particulate enzyme, with the former representing more than 50% of total at 48 h. The levels of gluconeogenic enzymes in the piglets in the early neonatal period would appear to be adequate for their needs and do not provide an explanation for their fasting hypoglycaemia. Hepatic levels of lactate, pyruvate, phosphoenolpyruvate, ketone bodies, and amino acids were determined in these piglets. No significant differences were observed in these metabolites between fasted and suckled animals except that glutamine was doubled in fed piglets, Evidence for the metabolic block in the livers of fasted animals was lacking and ketone bodies did not accumulate. These observations suggest that the limitations to gluconeogenesis result from unavailability of energy substrates and/or carbon precursors to the liver or the deficiency in their uptake.

Animals↗

Orotic acid content of infant formulas.

The orotic acid content of four commercially available infant formulas has been examined. Enfamil contains 118 microgram orotic acid per milliliter as fed, Similac 98, SMA 27, and Isomil less than 1 microgram/ml. As expressed relative to total solids, these formulas contain less than 0.1% orotic acid. Since consumption of 1% orotic acid does not lead to a fatty liver in any species examined other than the rat and 0.1% orotic acid fails to induce statistically significant hepatic changes in the rat, it is suggested that orotic acid at the level found in these formulas is not likely to pose a health hazard to the infants consuming them.

Infant Food↗

Orotic acid-induced metabolic changes in the rat.

The effects of orotic acid consumption on hepatic lipid and nucleotide metabolism were examined concurrently in the rat. When orotic acid was fed at 1.0%, fatty liver development was evident by day 7 but not by day 3, whereas a decreased purine/pyrimidine ratio of hepatic acid-soluble nucleotides was noted by day 3. As nucleotide changes preceded hepatic fat accumulation, they could be a contributing factor in the etiology of fatty liver. When fed for 10 days, rats did not accumulate hepatic fat on a diet containing 0.1%, but did on 0.5 and 1.0%; similarly, the purine/pyrimidine ratio was not significantly affected on 0.1% but was decreased on both 0.5 and 1.0% orotic acid. These results indicate that orotic acid alters both hepatic lipid and nucleotides when consumed by rats at a level above 0.1%. As measured in these studies, hepatomegaly did not always accompany hepatic steatosis and appeared to be a subsequent development. In addition, L-dihydroorotate was shown to be as effective as orotic acid in increasing hepatic weight and lipid whereas uridine was ineffective.

Animals↗

Superiority of 24-hour outpatient monitoring over multi-stage exercise testing for the evaluation of syncope.

Twenty-four hour outpatient monitoring was compared with maximum multi-stage Treadmill exercise testing for the detection of dysrhythmias producing syncope or near syncope in 119 patients. All patients had cardiovascular disorders; only 21% had coronary artery disease. Patients with obvious cause of syncope and/or significant dysrhythmias by resting electrocardiograms (ECGs) were excluded. A significant dysrhythmia was found which was considered the probable cause of syncope or pre-syncope in 76 patients (64%). A non-cardiac cause was found in tachycardia, exercise testing identified only patients with complex ventricular dysrhythmias and missed 64% of these. Overall, dysrhythmias were found by ambulatory monitoring alone in 63 patients, by exercise testing alone in only three, and by both in 10. It is concluded that: 1) in the majority of patients with syncope or pre-syncope suspected to be of a cardiac origin, a dysrhythmia can be found; 2) ambulatory monitoring is far superior to exercise testing for detection of dysrhythmias; 3) exercise testing increases the yield for complex ventricular dysrhythmias.

Adolescent↗

Histology and ultrastructure of the carotid sinus in experimental hypertension.

Histological studies including electron microscopy were performed on the carotid sinuses of dogs with renal hypertension of 17 to 82 days duration or hypertension of two years secondary to aortic coarctation. Physiological abnormalities in the baroreceptors including resetting had been demonstrated before death. Histologically there were no qualitative or quantitative differences in the intrasinus nerve fibers of the hypertensive animals compared tp controls with normal blood pressures, and there was no evidence that baroreceptor degeneration occurred as a consequence of the hypertension. Structural lesions confined to the intima were evident in the sinus walls of some of the renal hypertensive animals whereas the dogs with coarctation showed more advanced sinus wall changes including medial calcification and enlargement of the vascular lumen.

Animals↗

The active site of rabbit muscle pyruvate kinase. Evidence for a site common to the oxalacetate decarboxylase and pyruvate kinase reactions.

Rabbit muscle pyruvate kinase has been shown to catalyze the decarboxylation of oxalacetate (Creighton, D.J. and Rose, I.A. (1976) J. Biol. Chem. 251, 61). Noncovalent and covalent modifiers of the enzyme have been used to assess whether the decarboxylase and kinase reactions take place at a common site. Phosphoenol-alpha-ketobutyrate, an analog of the substrate phosphoenol-pyruvate, inhibits decarboxylase and kinase competitively and with nearly identical Ki values (5.7 micron and 4.8 micron, respectively). Oxalate, an analog of enol-pyruvate, inhibits each competitively and with similar Ki values (11 micron and 4.7 micron, respectively). Both activities are lost in parallel upon reaction with dithionitrobenzoate, which is active-site specific. These results indicate that the two activities share a common site on the enzyme. But, effects of the following modifiers suggest that different amino acid residues at that site participate in the two reactions: phenylalanine inhibition and fructose 1,6-bisphosphate activation are more effective with the decarboxylase; iodoacetamide preferentially inactivates decarboxylase while trinitrobenzenesulfonate preferentially inactivates kinase.

Animals↗

Accelerographic, temporal, and distance gait factors in below-knee amputees.

Gait characteristics of 19 patients with a unilateral below-knee amputation were studied. The accelerographic and foot placement method used in this study allowed for simultaneous acquisition of data commonly obtained in the experimental laboratory (acceleration) and data easily gathered in the physical therapy clinic (temporal and distance factors). The following results may be of interest to the clinician: 1) measures of cadence, stride length, and velocity were highly related and the magnitude of these measures was below commonly accepted values for normal; 2) the below-knee amputees spent more time in stance phase on their uninvolved lower extremity than on their involved (prosthetic) extemity; 3) the step length from heel strike of the uninvolved lower extremity to heel strike of the involved (prosthetic) lower extremity was greater and accomplished in less time than the opposite step; and 4) smoothness of the gait pattern and any single temporal and distance factor exhibited low statistical relationships.

Acceleration↗

Secondary kinase reactions catalyzed by yeast pyruvate kinase.

1. Yeast pyruvate kinase (EC 2.7.1.40) catalyzes, in addition to the primary, physiologically important reaction, three secondary kinase reactions, the ATP-dependent phosphorylations of fluoride (fluorokinase), hydroxylamine (hydroxylamine kinase) and glycolate (glycolate kinase). 2. These reactions are accelerated by fructose-1,6-bisphosphate, the allosteric activator of the primary reaction. Wth Mg2+ as the required divalent cation, none of these reactions are observed in the absence of fructose-biphosphate. With Mn2+, fructose-bisphosphate is required for the glycolate kinase reaction, but merely stimulates the other reactions. 3. The effect of other divalent cations and pH on three secondary kinase reactions was also examined. 4. Results are compared with those obtained from muscle pyruvate kinase and the implications of the results for the mechanism of the yeast enzyme are discussed.

Adenosine Triphosphate↗

The proton transfer reactions catalyzed by yeast pyruvate kinase.

1. The proton-transfer reactions of yeast pyruvate kinase (EC 2.7.1.40) were studied. Proton-transfer from C-3 of phosphoenolpyruvate to water occurs only in the presence of the phosphoryl-acceptor ADP. Proton transfer from C-3 of pyruvate to water occurs only in the presence of ATP. However, the proton transfer in the latter case occurs 10-100 times faster than phosphoryl transfer; this supports a mechanism in which proton transfer precedes phosphoryl transfer in the reverse reaction of pyruvate kinase. 2. The characteristics of proton-transfer reactions of yeast pyruvate kinase were compared with those previously reported for rabbit muscle pyruvate kinase (Robinson, JL. and Rose, I.A. (1972) J. Biol. Chem. 247, 1096-1105). The pH-profiles and the divalent cation dependencies were similar for Fru-1,6-P2-activated yeast pyruvate kinase and the muscle enzyme. Pyruvate enolization by yeast pyruvate kinase has an absolute requirement for ATP in contrast to enolization by the muscle enzyme which proceeds when ATP is replaced by Pi or other dianions. 3. Fructose-1,6-bisphosphate was shown to affect the catelytic steps of yeast pyruvate kinase in addition to the binding of substrates. Its role depends on the divalent cation used to activate the enzyme.

Adenosine Diphosphate↗