Biomedical subjects
Publications and source records attributed to J M Miles.
When performing a sensory island transfer for restoration of sensibility to an area of pinch, one must be accurate in determining the recipient site. We present a simple and precise means of determining the recipient site for a sensory island transfer by ink-blotting the areas of contact of the thumb and index finger.
Infusion of epinephrine in humans increases glucose production and decreases plasma concentrations of some essential amino acids such as leucine, while not affecting the plasma concentration of the potential gluconeogenic amino acid alanine. To determine whether epinephrine alters alanine and leucine metabolism, rates of appearance (Ra) and disappearance (Rd) of glucose, alanine, and leucine were determined in postabsorptive volunteers using [3H]glucose, [2H3]alanine, [15N]leucine, and [2H3]leucine during a 180-min infusion of epinephrine (50 ng X kg-1 X min-1). Plasma glucose (90 +/- 1 to 142 +/- 5 mg/dl) and insulin (10 +/- 1 to 16 +/- 2 micrograms/ml) increased (P less than 0.05), whereas plasma alanine concentrations did not change and plasma leucine concentrations increased (127 +/- 5 to 72 +/- 3 microM). Glucose Ra increased transiently and returned to basal values by 120 min. In contrast, alanine Ra and Rd increased identically and progressively from 5.7 +/- 0.5 to 14.5 +/- 1.9 mumol X kg-1 X min-1 by 180 min. Although leucine nitrogen Ra increased transiently and returned to basal values, leucine carbon Ra and Rd decreased (P less than 0.05) during the infusion of epinephrine. The calculated rate and percent of leucine nitrogen going to alanine increased, whereas the percent of alanine nitrogen derived from leucine remained constant. The increase in alanine Ra was entirely attributable to increased de novo synthesis because proteolysis, as estimated by leucine carbon flux, decreased.
Prolonged exposure to glucocorticoids in pharmacologic amounts results in muscle wasting, but whether changes in plasma cortisol within the physiologic range affect amino acid and protein metabolism in man has not been determined. To determine whether a physiologic increase in plasma cortisol increases proteolysis and the de novo synthesis of alanine, seven normal subjects were studied on two occasions during an 8-h infusion of either hydrocortisone sodium succinate (2 micrograms/kg X min) or saline. The rate of appearance (Ra) of leucine and alanine were estimated using [2H3]leucine and [2H3]alanine. In addition, the Ra of leucine nitrogen and the rate of transfer of leucine nitrogen to alanine were estimated using [15N]leucine. Plasma cortisol increased (10 +/- 1 to 42 +/- 4 micrograms/dl) during cortisol infusion and decreased (14 +/- 2 to 10 +/- 2 micrograms/dl) during saline infusion. No change was observed in plasma insulin, C-peptide, or glucagon during either saline or cortisol infusion. Plasma leucine concentration increased more (P less than 0.05) during cortisol infusion (120 +/- 1 to 203 +/- 21 microM) than saline (118 +/- 8 to 154 +/- 4 microM) as a result of a greater (P less than 0.01) increase in its Ra during cortisol infusion (1.47 +/- 0.08 to 1.81 +/- 0.08 mumol/kg X min for cortisol vs. 1.50 +/- 0.08 to 1.57 +/- 0.09 mumol/kg X min). Leucine nitrogen Ra increased (P less than 0.01) from 2.35 +/- 0.12 to 3.46 +/- 0.24 mumol/kg X min, but less so (P less than 0.05) during saline infusion (2.43 +/- 0.17 to 2.84 +/- 0.15 mumol/kg X min, P less than 0.01). Alanine Ra increased (P less than 0.05) during cortisol infusion but remained constant during saline infusion. During cortisol, but not during saline infusion, the rate and percentage of leucine nitrogen going to alanine increased (P less than 0.05). Thus, an increase in plasma cortisol within the physiologic range increases proteolysis and the de novo synthesis of alanine, a potential gluconeogenic substrate. Therefore, physiologic changes in plasma cortisol play a role in the regulation of whole body protein and amino acid metabolism in man.
Four experiments are reported in which subjects vocalized words presented in the visual half-fields in a baseline period. Then two of the words were followed by aversive noise in a punishment phase, and finally, the baseline procedure was repeated. In two studies, as predicted, right handed males' reaction time improved least on words associated with the punished words when they appeared in the left visual field and had to cross from right to left hemisphere to be vocalized. The effect was not found among female subjects in the third study. Finally, the effect was found only among subjects in the highest third on the Repression-Sensitization scale. Taken together, the studies support the notion of selective inhibition of aversive processes that must cross from right to left hemispheres, although individual differences limit the generalizability of this statement.
The hyperglycaemia and hyperketonaemia of diabetic ketoacidosis are initiated primarily by overproduction of these substrates; subsequent maintenance of hyperglycaemia occurs, in large part, due to impaired utilization of glucose, whereas overproduction of ketone bodies continues to be the major mechanism for maintenance of hyperketonaemia. Insulin deficiency results in increased rates of lipolysis and provides increased substrate (free fatty acids) for ketogenesis. Hyperglucagonaemia can augment ketogenesis further in the setting of insulin deficiency. It is likely that other counter-insulin hormones (growth hormone, catecholamines) also contribute to the pathogenesis of DKA, though their role is less well defined. Insulin corrects DKA largely via suppression of lipolysis (and thus ketone body production); insulin suppresses glucose production at lower levels than it does ketone body production.
The present studies were undertaken to assess the relative effects of free fatty acid (FFA) availability, glucagon excess, and insulin deficiency on ketone body (KB) production in man. To determine whether an increase in FFA availability would augment KB production in the absence of insulin deficiency and glucagon excess, plasma insulin and glucagon were maintained at basal concentrations by infusion of somatostatin and exogenous insulin and glucagon, and plasma FFA were increased from 0.32 +/- 0.06 to 1.4 +/- 0.1 mM by a 2.5-h-infusion of a triglyceride emulsion plus heparin. KB production increased fivefold from 2.2 +/- 0.4 to 11.4 +/- 1.2 mumol . kg-1 . min-1, P less than 0.001. To determine whether insulin deficiency would further augment KB production, analogous experiments were performed but the replacement infusion of insulin was stopped. Despite a greater increase in plasma FFA (from 0.26 +/- 0.04 to 1.95 +/- 0.3 mM), KB production increased (from 1.5 +/- 0.3 to 11.1 +/- 1.8 mumol . kg-1 . min-1) to the same extent as in the absence of insulin deficiency. To determine whether hyperglucagonemia would augment KB production beyond that accompanying an increase in plasma FFA and, if so, whether this required insulin deficiency, similar experiments were performed in which the glucagon infusion rate was increased to produce plasma glucagon concentrations of 450-550 pg/ml with and without maintenance of the basal insulin infusion. When basal plasma insulin concentrations were maintained, hyperglucagonemia did not further increase KB production; however, when the basal insulin infusion was discontinued, hyperglucagonemia increased KB production significantly, whereas no change was observed in saline control experiments. These studies indicate that, in man, FFA availability is a major determinant of rates of KB production; insulin does not appear to influence ketogenesis rates by a direct hepatic effect, and glucagon can further augment KB production when FFA concentrations are increased but only in the setting of insulin deficiency.
Sixteen cases of osteosclerotic myeloma and peripheral neuropathy (SM-PN) were reviewed. The neuropathy resembled chronic inflammatory-demyelinating polyneuropathy with predominantly motor disability, high CSF protein levels, and low motor nerve conduction velocities. Twelve of the 16 patients had detectable levels of monoclonal serum proteins, all with lambda light chains, but results of other laboratory studies were usually normal. Most of the patients also had organomegaly, endocrine abnormalities, or both. Treatment of solitary lesions with tumoricidal irradiation usually improved the neuropathy and reversed the nonneurologic abnormalities; chemotherapy for multiple osteosclerotic lesions was less helpful.
Ketone bodies have been suggested to have a protein-sparing effect, since infusion of Na-beta-hydroxybutyrate in man decreases plasma alanine concentrations and urinary nitrogen (N) excretion. To test this hypothesis, six normal postabsorptive volunteers were infused with Na-beta-hydroxybutyrate for 3 h. Rates of glucose, leucine carbon, and alanine appearance and disappearance from the plasma space were traced with [3-3H]glucose, L-[6,6,6-2H3]leucine, and [2,3,3,3-2H4]alanine. Rates of leucine N appearance and disappearance and the rate of transfer of leucine N to alanine were assessed with [15N]leucine. During ketone body infusion, plasma alanine decreased (P less than 0.05), whereas plasma leucine increased (P less than 0.05). Rates of alanine appearance increased (5.3 +/- 0.3 to 7.8 +/- 0.6 mumol/kg X min), but the increase in its rate of disappearance was slightly greater, accounting for the decrease in plasma alanine concentration. Leucine N flux and the rate and percent of leucine N transferred to alanine increased, whereas leucine carbon flux was unchanged. To determine the effect of the alkalemia induced by Na-beta-hydroxybutyrate, four additional subjects were infused with NaHCO3. Alkalemia had no effect on leucine N or carbon flux or on the rate of appearance of alanine, but increased the rate of alanine disappearance, resulting in a decrease in the plasma alanine concentration. Since the rate of appearance of leucine carbon was unaltered during the infusion of Na-beta-hydroxybutyrate, it is unlikely that hyperketonemia per se decreases proteolysis in postabsorptive man.
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In normal human subjects, when plasma insulin, glucagon and growth hormone were 'clamped' at basal concentrations (by infusion of somatostatin plus replacement infusion of these hormones), infusion of Intralipid and heparin increased plasma free fatty acid (FFA) concentrations to approx. 1.3 mM, and ketone body production increased 4-5 fold to approx. 11 mumol . kg -1 . min-1. Hyperglucagonaemia did not further increase ketogenesis. In conditions of combined insulin and glucagon deficiency (by infusion of somatostatin without insulin and glucagon), administration of Intralipid and heparin increased plasma FFA concentrations to approx. 2.2 mM but a further increase in ketone body production did not accompany this increase. In these conditions hyperglucagonaemia increased ketogenesis by 2-3 fold the increment seen in control studies. Infusion of adrenaline (epinephrine) in conditions in which insulin secretion was not inhibited caused only a transient increase in plasma FFA concentrations and in ketone body production. These data indicate: (1) that in humans increased FFA availability can markedly augment ketogenesis in the absence of insulin deficiency and without hyperglucagonaemia; (2) that glucagon can increase ketone body production during insulin deficiency but not in its absence; and (3) that insulin deficiency may be accompanied by increased ketogenesis only because of a lack of its restraint on lipolysis and because of the action of glucagon. Glucagon may be important in determining the magnitude of ketone body production for a given degree of FFA availability and insulin deficiency, and may be necessary for attainment of maximal rates of ketogenesis. Adrenaline increases ketone body production in humans, but whether this is primarily due to a direct effect on the liver or is mediated through enhancement of lipolysis remains to be determined.
The importance of alpha-keto acid binding to plasma proteins was investigated both in vitro and in vivo using alpha-ketoisocaproate (KIC), the alpha-keto acid of leucine. Gel chromatography indicated that 65% of the radioactivity comigrated with serum albumin. An ultrafiltration assay was developed to estimate the percentage of free and bound KIC. These percentages, along with total plasma KIC concentrations, were used to calculate the circulating concentrations of free and bound KIC. KIC or free fatty acids (FFA) displaced [14C]KIC bound to bovine albumin or whole plasma. KIC was totally displaced from plasma proteins by 10 mM oleate, stearate, and myristate; whereas the alpha-keto acids of isoleucine and value were 50 and 85%, respectively, as effective as KIC. To determine whether increased plasma FFA concentrations alter the binding of KIC to plasma proteins in vivo, five postabsorptive humans were infused with triglyceride and heparin during the simultaneous administration of somatostatin, glucagon, and insulin. During the FFA elevation, plasma leucine decreased by 9% (P less than 0.02). Total plasma KIC remained constant, whereas free KIC increased (P less than 0.02) and bound KIC decreased (P less than 0.001). These results indicate that KIC is bound to plasma albumin in vivo and suggests that FFA, by altering circulating free KIC concentrations, may influence protein metabolism in man.
In vitro perfusion and incubation studies and recent investigations in dogs suggest that branched chain amino acids (BCAA) may be a major source of alanine nitrogen. To determine the contribution of BCAA nitrogen to the formation of alanine in man, seven postabsorptive adults received prime-dose constant infusions of 15N-leucine, L-[6,6,6-2H3] leucine, and L-[2,3,3,3-2H4] alanine; isotopic enrichment was determined in arterialized venous plasma samples by gas chromatography-mass spectroscopy. At substrate and isotope steady state, alanine flux and the rate of 15N alanine appearance were 5.4 +/- 0.3 mumol/kg-min and 32 +/- 2 nmol/kg.min, respectively. Leucine nitrogen flux was significantly greater than that of leucine carbon flux (2.54 +/- 0.25 vs. 1.90 +/- 0.10 mumol/kg.min, respectively; P less than 0.001). The 30% greater flux of leucine nitrogen when compared with leucine carbon suggests significant recycling of the leucine carbon in vivo. The percent of circulating alanine nitrogen derived from leucine was 12.5 +/- 1.5%; however, the rate of leucine nitrogen transferred to alanine was 0.66 +/- 0.05 mumol/kg.min, and represents a minimum of 28% of leucine nitrogen going to alanine. On the basis of these data, together with the percent of alanine and leucine in body protein, only 40% of circulating plasma alanine could come from endogenous protein, whereas 60% is derived from de novo synthesis. In addition, at least 20% of the nitrogen required for alanine synthesis is derived solely from leucine following an overnight fast. Therefore, if the contribution of isoleucine and valine nitrogen is similar to that of leucine, the BCAA may contribute to a minimum of 60% of the nitrogen required for alanine synthesis in postabsorptive man.
Myosins and myosin light chain kinases have been isolated from a cloned line of myoblasts (L5/A10) as this cell line undergoes differentiation toward adult muscle. At least three myosin isozymes were obtained during this developmental process. Initially a nonmuscle type of myosin was found in the myoblasts. The molecular weights of the myoblast light chains were 20 000 and 15 000. Myosin isolated from early myotubes had light chains with molecular weights of 20 000 and 19 500. Myosin isolated from myotubes which contained sarcomeres had light chains with molecular weights of 23 000, 18 500, and 16 000. This last myosin was similar in light chain complement to adult rat thigh muscle. Two forms of the myosin light chain kinase activity were detected: a calcium-independent kinase in the myoblasts and a calcium-dependent kinase in the myotubes with sarcomeres. No myosin light chain kinase activity was detected in the early myotubes.
To determine the mechanism by which ketone bodies decrease plasma glucose in man, seven normal postabsorptive volunteers were infused for 3 h with beta-hydroxybutyrate. Total plasma ketone bodies (beta-hydroxybutyrate plus acetoacetate) increased to levels (approximately 2.5 mM) observed after a 2- to 3-day fast in normal subjects. Plasma glucose decreased 10% concomitant with decreases of 25% and 10%, respectively, in the rates of glucose production and glucose utilization determined isotopically with [3-3H]glucose. Plasma insulin and glucagon concentrations were unaltered, but plasma C-peptide levels increased from 2.6 +/- 0.1 ng/ml to a maximum of 3.9 +/- 0.2 ng/ml at 30 min (P < 0.01) and remained significantly increased for more than 2 h. Plasma alanine decreased approximately 14% (P < 0.05), while plasma lactate increased 25% (P < 0.01) so that there was no net decrease in the combined levels of these gluconeogenic substrates. These results demonstrate that physiological increments in circulating ketone body concentrations decrease plasma glucose in normal man by suppressing glucose production, an effect which can be explained by the stimulation of insulin secretion being reflected only in changes in plasma C-peptide. Thus, changes in pancreatic B cell function not sufficient to alter peripheral plasma insulin levels may cause significant changes in hepatic glucose production.
The records of 20 patients with typical or sclerotic (sclerotic or mixed sclerotic and lytic) multiple myeloma and peripheral neuropathy who were seen during a 13-year period were analyzed to determine the natural history of the neuropathy. In the 10 patients with typical multiple myeloma, the clinical pattern was heterogeneous. Chemotherapy for multiple myeloma did not affect the neuropathy. The 10 patients who had sclerotic myeloma presented in a relatively homogeneous manner, with a predominantly motor neuropathy. Tumoricidal radiation therapy to localized plasmacytomas resulted in pronounced improvement in the neuropathy of two patients, mild improvement in three, and mild improvement in the endocrine abnormalities of one patient.
To determine the mechanism by which hyperinsulinemia causes hypoglycemia in insulinoma patients, rates of glucose production and utilization, and circulating levels of insulin, glucagon, alanine, lactate, and glycerol were measured in 6 insulinoma patients during development of fasting hypoglycemia and in 8 normal volunteers studied over an identical interval. Initially, insulinoma patients had a greater plasma insulin (42 +/- 9 versus 15 +/- 1 microunits/ml) and glucagon levels (214 +/- 31 versus 158 +/- 21 pg/ml) than normal subjects, P less than 0.05, but their plasma glucose levels (81 +/- 4 mg/dl) and rates of glucose production and utilization (1.71 +/- 0.08 and 1.74 +/- 0.08 mg/kg . min, respectively) were not significantly different from those of normal subjects (93 +/- 2 mg/dl, 1.93 +/- 0.11, and 1.92 +/- 0.13 mg/kg . min, respectively). During a subsequent 8-h fast, glucose production and glucose utilization decreased in both groups, but more markedly in insulinoma patients. Since glucose utilization exceeded glucose production to a greater extent in insulinoma patients than in normal subjects, plasma glucose decreased to 44 +/- 3 mg/dl in insulinoma patients, but only to 84 +/- 1 mg/dl in normal subjects (P less than 0.001). Glucose utilization in insulinoma patients never exceeded that of normal subjects. These results demonstrate that fasting hypoglycemia in the insulinoma patients is usually due to suppression of glucose production rather than to acceleration of glucose utilization, as is widely thought. A direct effect of insulin on the liver is probably responsible, since circulating levels of gluconeogenic precursors are normal and since plasma glucagon increases during development of hypoglycemia in insulinoma patients.
A simple and reliable method is described for the determination of leucine flux in vivo using two stable isotopes of leucine and gas chromatography-mass spectrometry (GC-MS). [6,6,6-2H3]Leucine is administered as a primed-dose constant infusion in vivo and DL-[2H7]-leucine is added to plasma as an internal standard. Plasma leucine concentration and moles per cent enrichment of [2H3]leucine can be determined simultaneously by GC-MS and selected ion monitoring. Leucine flux calculated from the [6,6,6-2H3]leucine data was nearly identical to that obtained with L-[U-14C]leucine in dogs. This method is readily applicable to the study of leucine metabolism in humans of all ages and laboratory animals.
Epinephrine (50 ng . kg-1 . min-1) was infused for 120 min in seven normal volunteers alone (combined alpha- and beta-adrenergic stimulation), with propranolol (alpha-adrenergic stimulation), and with propranolol plus phentolamine (alpha-adrenergic blockade superimposed on alpha-adrenergic stimulation). During alpha-adrenergic stimulation, plasma glucose and glucose production increased 32 and 42% less, respectively, than during infusion of epinephrine alone, whereas glucose clearance was suppressed comparably. Plasma insulin decreased during alpha-adrenergic stimulation but increased during infusion of epinephrine alone. Plasma epinephrine was threefold greater during infusion of epinephrine plus propranolol than during infusion of epinephrine alone. When alpha-adrenergic blockade was superimposed on alpha-adrenergic stimulation, the increases in plasma glucose and glucose production as well as the decreases in plasma insulin and glucose clearance observed during alpha-adrenergic stimulation were virtually abolished, whereas plasma epinephrine levels were unaltered. These results indicate that in man epinephrine can cause hyperglycemia via both alpha- and beta-adrenergic stimulation of glucose production and suppression of glucose clearance, either directly or indirectly. alpha-Adrenergic effects on glucose production and clearance may be mediated by inhibition of insulin secretion.