Acupuncture in children and adolescents with bronchial asthma: a randomised controlled study.

OBJECTIVE: This randomised controlled trial evaluates the immediate effects of acupuncture as an add-on therapy in in-patient rehabilitation of children and adolescents with bronchial asthma. PATIENTS AND METHODS: In a pre-post design, the severity of symptoms, lung function, illness-specific quality of life (Paediatric Asthma Quality of Life Questionnaire - PAQLQ) and general and asthma-specific level of anxiety (State-Trait Anxiety Inventory for Children - STAIC) were investigated in 46 acupuncture and 47 control patients. In addition to asthma sports, climate therapy and behavioural training, the intervention group received acupuncture treatment with a standardised needle pattern (12 × 30 min.). RESULTS: With acupuncture, the peak expiratory flow variability differs significantly (p<0.01) from that of the control patients' group. Moreover, the acupuncture group differs significantly in their rehabilitation response at the time of discharge concerning perceived anxiety (STAIC-S). The lung function tests do not present differences between groups. CONCLUSION: After additional acupuncture, amelioration of peak expiratory flow variability and anxiety can be shown, without any difference in objective lung function tests and quality of life between study groups. Further studies might evaluate the effects of acupuncture on childhood asthma in an outpatient setting.

Expression profile of FSHD supports a link between retinal vasculopathy and muscular dystrophy.

BACKGROUND: Facioscapulohumeral muscular dystrophy (FSHD) is caused by deletions within a tandem array of D4Z4 repeats on chromosome 4q35. In addition to muscle degeneration, most patients with FSHD develop abnormalities of the retinal vasculature. Previous work has suggested that muscle degeneration in FSHD results from increased expression of genes proximal to the deletion, including FRG1. OBJECTIVES: To reexamine this mechanism and identify pathways that are abnormally regulated early in the disease process. METHODS: We prospectively studied gene expression in skeletal muscle in patients with FSHD (n = 19) vs healthy individuals (n = 30) and patients with myotonic dystrophy type 1 (n = 12). We used oligonucleotide microarrays for global analysis of gene expression and reverse transcriptase-PCR (RT-PCR) to assess expression or alternative splicing for particular genes. RESULTS: Expression of FRG1 was not increased in patients with FSHD, either by microarray analysis or quantitative RT-PCR. Among genes on 4q35, only LRP2BP showed upregulation that was specific to FSHD. However, neither LRP2BP nor FRG1 showed imbalance of allelic expression by RT-PCR. After filtering out genes that showed similar dysregulation in other forms of muscular dystrophy, only 44 genes were specifically upregulated early in FSHD. Among these, 34 genes were characterized or partially characterized, of which 11 (32%) had a role in vascular smooth muscle or endothelial cells. CONCLUSION: Expression of genes on chromosome 4q35 was normally regulated in the early stages of facioscapulohumeral muscular dystrophy. Our results support a possible link between muscular dystrophy and retinal vasculopathy in facioscapulohumeral muscular dystrophy.

Senescence-related changes in gene expression in muscle: similarities and differences between mice and men.

A microarray study of the effect of senescence in mice on gene expression in muscle has been published recently. The present analysis was done to evaluate the extent to which the age-related differences in gene expression in murine muscle are also evident in human muscle. RNA extracted from muscle of young (21-24 yr) and old men (66-77 yr) was studied both by serial analysis of gene expression (SAGE) and by oligonucleotide microarrays. SAGE tags were detected for 61 genes homologous to genes reported to be differentially expressed in young and old murine muscle. The microarray had probe sets for 70 homologous genes. For 17 genes, there was evidence for a similar age-related change in expression in muscles of mice and men. For 32 other genes, there was evidence that the effect of age on the level of expression is not the same in mice and men. There was no evidence that older human muscle has increased expression of the stress response genes that are increased in old murine muscle.

High-abundance mRNAs in human muscle: comparison between young and old.

To gain a better understanding of the potential role of altered gene expression in the diminished muscle function in old age, we performed a broad search for transcripts expressed at quantitatively different levels in younger (21-24 yr) and older (66-77 yr) human vastus lateralis muscle by serial analysis of gene expression (SAGE). Because SAGE was based on RNA pooled from muscle of several different subjects, relative concentrations of selected mRNAs also were determined in individual muscle samples by quantitative RT-PCR. There were 702 SAGE tags detected at least 10 times in one or both mRNA pools, and the detection frequency was different (at P < 0.01) between young and older muscle for 89 of these. The ratio of myosin heavy chain 2a mRNA to myosin heavy chain 1 mRNA was reduced in older muscle. The mRNAs encoding several mitochondrial proteins involved in electron transport (including several subunits of cytochrome-c oxidase and NADH dehydrogenase) and subunits of ATP synthase were approximately 30% less abundant in older muscle. Several mRNAs encoding enzymes involved in glucose metabolism also were less abundant in older muscle. Analysis of individual samples revealed that the differences suggested by SAGE were not artifacts of atypical gene expression in one or a few individuals. These data suggest that some of the phenotypic changes in senescent muscle may be related to altered gene transcription.

Expanded CUG repeat RNAs form hairpins that activate the double-stranded RNA-dependent protein kinase PKR.

Myotonic dystrophy is caused by an expanded CTG repeat in the 3' untranslated region of the DM protein kinase (DMPK) gene. The expanded repeat triggers the nuclear retention of mutant DMPK transcripts, but the resulting underexpression of DMPK probably does not fully account for the severe phenotype. One proposed disease mechanism is that nuclear accumulation of expanded CUG repeats may interfere with nuclear function. Here we show by thermal melting and nuclease digestion studies that CUG repeats form highly stable hairpins. Furthermore, CUG repeats bind to the dsRNA-binding domain of PKR, the dsRNA-activated protein kinase. The threshold for binding to PKR is approximately 15 CUG repeats, and the affinity increases with longer repeat lengths. Finally, CUG repeats that are pathologically expanded can activate PKR in vitro. These results raise the possibility that the disease mechanism could be, in part, a gain of function by mutant DMPK transcripts that involves sequestration or activation of dsRNA binding proteins.

Inventory of high-abundance mRNAs in skeletal muscle of normal men.

G42875rial analysis of gene expression (SAGE) method was used to generate a catalog of 53,875 short (14 base) expressed sequence tags from polyadenylated RNA obtained from vastus lateralis muscle of healthy young men. Over 12,000 unique tags were detected. The frequency of occurrence of each tag reflects the relative abundance of the corresponding mRNA. The mRNA species that were detected 10 or more times, each comprising >/=0.02% of the mRNA population, accounted for 64% of the mRNA mass but <10% of the total number of mRNA species detected. Almost all of the abundant tags matched mRNA or EST sequences cataloged in GenBank. Mitochondrial transcripts accounted for approximately 20% of the polyadenylated RNA. Transcripts encoding proteins of the myofibrils were the most abundant nuclear-encoded mRNAs. Transcripts encoding ribosomal proteins, and those encoding proteins involved in energy metabolism, also were very abundant. The database can be used as a reference for investigations of alterations in gene expression associated with conditions that influence muscle function, such as muscular dystrophies, aging, and exercise.

Stimulation of myofibrillar synthesis by exercise is mediated by more efficient translation of mRNA.

Resistance exercises stimulate protein synthesis in human muscle, but the roles of changes in mRNA concentrations and changes in the efficiency of mRNA translation have not been defined. The present study was done to determine whether resistance exercise affects concentrations of total RNA, total mRNA, actin mRNA, or myosin heavy-chain mRNA (total and isoform specific). Eight subjects, 62-75 yr old, performed unilateral knee extensions at 80% of their one-repetition-maximum capacity on days 1, 3, and 6 of the study. On day 7, biopsies of exercised and nonexercised vastus lateralis muscles were obtained. Myofibrillar synthesis was determined by stable- isotope incorporation, and mRNA concentrations were determined by membrane hybridization and PCR-based methods. The exercise stimulated myofibrillar synthesis [30 +/- 6 (SE)%] without affecting RNA or mRNA concentrations. The effect of exercise on protein synthesis in individual subjects did not correlate with the effect on total RNA and mRNA concentrations. These data suggest that the stimulation of myofibrillar synthesis by resistance exercise is mediated by more efficient translation of mRNA.

Human kidney and liver gluconeogenesis: evidence for organ substrate selectivity.

To assess the contribution of the human kidney to gluconeogenesis (GN) and its role in conversion of glutamine and alanine to glucose, we used a combination of isotopic and organ balance techniques in nine normal postabsorptive volunteers and measured both overall and renal incorporation of these precursors into glucose before and after infusion of epinephrine. In the postabsorptive basal state, renal incorporation of glutamine (27 +/- 2 mumol/min) and alanine (2.1 +/- 0.5 mumol/min) into glucose accounted for 72.8 +/- 3.3 and 3.9 +/- 0.5% of their overall incorporation into glucose (37 +/- 2 and 51 +/- 6 mumol/min, respectively) and 19.0 +/- 3.5 and 1.4 +/- 0.2%, respectively, of overall renal glucose release. Infusion of epinephrine, which increased systemic and renal glucose release more than twofold (P < 0.001), increased overall glutamine and alanine incorporation into glucose (both P < 0.001) and increased renal GN from glutamine (P < 0.001) but not from alanine (P = 0.15). Renal glutamine GN now accounted for 90.3 +/- 4.0% of overall glutamine GN (P = 0.01 vs. basal), whereas renal alanine GN still accounted for only 4.8 +/- 1.7% of overall alanine GN (P = 0.36 vs. basal). With the assumption that kidney and liver are the only gluconeogenic organs in humans, these results indicate that glutamine GN occurs primarily in kidney, whereas alanine GN occurs almost exclusively in liver. Isotopic studies of glutamine and alanine incorporation into plasma glucose may provide a selective, noninvasive method to assess hepatic and renal GN.

High-protein meals do not enhance myofibrillar synthesis after resistance exercise in 62- to 75-yr-old men and women.

This study tested the hypothesis that increasing the protein content of isocaloric meals increases the rate of myofibrillar synthesis in muscle of healthy subjects over 60 yr old and enhances the stimulation of myofibrillar synthesis induced by resistance exercise. Myofibrillar synthesis of sedentary and exercised quadriceps muscle was determined by incorporation of L-[1-13C]leucine. During the tracer infusion, subjects consumed meals with a low (7% of energy, n = 6)-, normal (14%, n = 6)-, or high (28%, n = 6)-protein content. In sedentary muscle, the mean (+/- SE) myofibrillar synthesis was 1.56 +/- 0.13%/day in the low-protein group, 1.73 +/- 0.11 %/day in the normal-protein group, and 1.76 +/- 0.10%/day in the high-protein group (P = 0.42). Myofibrillar synthesis was faster in exercised muscle (mean 27%, P < 10(-6) in all groups (2.10 +/- 0.14%/day in low protein; 2.18 +/- 0.10%/day in normal protein; 2.11 +/- 0.09%/day in high protein; P = 0.84). The stimulation of myofibrillar synthesis by exercise was not significantly different among low-protein [0.54 +/- 0.12%/day (37 +/- 9%)], normal-protein [0.46 +/- 0.08%/day (28 +/- 5%)], and high-protein groups [0.34 +/- 0.04%/day (20 +/- 3%); P = 0.31]. We conclude that high-protein meals do not enhance the stimulation of myofibrillar protein synthesis induced by resistance exercise.

Growth hormone and insulin-like growth factor-I as anabolic agents.

The reduced growth hormone and insulin-like growth factor-I concentrations in growth hormone deficiency and normal ageing are associated with reduced muscle mass and strength, and slower muscle protein synthesis. Recent research has addressed the hypothesis that growth hormone and insulin-like growth factor-I have an anabolic effect in adults, including the elderly. These hormones stimulate whole-body and muscle protein synthesis, at least under some conditions. There is increasing evidence to justify long-term administration of growth hormone to promote muscle growth in growth hormone deficient adults. However, the long-term effects on muscle mass and function in the elderly do not seem beneficial enough to justify widespread hormone replacement therapy. These hormones may be useful anabolic agents to counteract muscle wasting under other conditions, including surgical stress, renal failure, muscular dystrophy, glucocorticoid administration and HIV infection, but more clinical trials are needed to determine the functional significance of the protein anabolic effects under these conditions.

Insulin-like growth factor-I, actin, and myosin heavy chain messenger RNAs in skeletal muscle after an injection of growth hormone in subjects over 60 years old.

Growth hormone (GH) increases the amount of insulin-like growth factor-I (IGF-I) mRNA in rat skeletal muscle, but this effect has not been demonstrated in human muscle. An autocrine effect of IGF-I produced in muscle may be an important determinant of the increased muscle mass associated with GH therapy. Thus, we examined IGF-I mRNA abundance in skeletal muscle biopsy samples taken 10 h after a subcutaneous injection of GH (0.03 mg/kg, n = 6) or placebo (normal saline, n = 5) in men and women over 60 years of age. Relative tissue concentrations of IGF-I mRNA were evaluated with a competitive reverse transcriptase-polymerase chain reaction assay. Mean plasma IGF-I concentrations rose steadily after the GH injection, and were 74% higher in the GH group than in the control group at the time of the muscle biopsies. There was no consistent difference between the GH and control groups in muscle IGF-I mRNA abundance when expressed in relation to total RNA or polyadenylated RNA. However, one GH-treated subject had three times more IGF-I mRNA, relative to polyadenylated RNA, than the average control subject. There was no effect of GH on levels of mRNAs encoding the most abundant myofibrillar proteins, actin and myosin heavy chain. These data do not support the hypothesis that increased IGF-I mRNA abundance in skeletal muscle is required for the anabolic effect of GH in people over 60 years of age.

Expression of elongation factor-1 alpha and S1 in young and old human skeletal muscle.

Previous research has indicated that reduced expression of elongation factor-1 alpha (EF-1 alpha) may be an important determinant of the reduced rate of protein synthesis in senescent animals and cultured cells. The present study examined whether expression of EF-1 alpha or S1, a homologous protein found exclusively in postmitotic tissues, is reduced in senescent human skeletal muscle. Muscle biopsies were obtained from the vastus lateralis muscles of healthy young (22-31 yr old) and old (61-74 yr old) subjects. As reported previously, myofibrillar protein synthesis was approximately 40% slower in the older muscle (p < .001) as determined by incorporation of a stable isotope. Immunoblotting revealed no difference in the concentration of EF-1 alpha + S1 between younger and older muscle. RT-PCR assays indicated that S1 mRNA was much more abundant than EF-1 alpha mRNA in muscles of both age groups, with no reduction in either EF-1 alpha or S1 mRNA abundance in older muscles. We conclude that expression of EF-1 alpha and S1 is not diminished in older muscles and does not explain the age-related slowing of protein synthesis in human skeletal muscle. However, we cannot exclude the possibility that the activity of these proteins declines during senescence due to post-translational modifications.

Steady-state and non-steady-state measurements of plasma glutamine turnover in humans.

To compare steady-state glutamine turnover using nitrogen, carbon, and hydrogen tracers and to test the validity of monocompartmental equations to determine plasma glutamine turnover under non-steady-state conditions, we infused 10 normal postabsorptive volunteers simultaneously with [3,4-3H]glutamine, [2-15N]glutamine, and [U-14C]glutamine for 4 h to isotopic steady state. Eight of the ten subjects were subsequently infused in a stepwise fashion with exogenous glutamine. Plasma glutamine enrichment and specific activities fit a monoexponential model well (r = 0.89, 0.92, and 0.92 for [2-15N]-, [U-14C]-, and [3,4-3H]glutamine, respectively). Volumes of distribution for each tracer (362 +/- 58, 433 +/- 51, and 446 +/- 63 ml/kg) and the transfer rate constants (0.0224 +/- 0.0020, 0.0222 +/- 0.0020, and 0.0240 +/- 0.0023 min(-1)) for [2-15N]-, [U-14C]-, and [3,4-3H]glutamine, respectively, were not significantly different from one another. However, turnover of glutamine determined with [3,4-3H]glutamine (6.14 +/- 0.54 micromol x kg(-1) x min(-1)) exceeded that determined with [U-14C]glutamine (5.72 +/- 0.541 micromol x kg(-1) x min(-1); P < 0.03), which in turn exceeded that determined with [2-15N]glutamine (4.67 +/- 0.39 micromol x kg(-1) x min(-1), P < 0.01). The monocompartmental non-steady-state equations of both DeBodo et al. (DeBodo, R., R. Steele, A. Dunn, and J. Bishop. Rec. Prog. Horm. Res. 19: 445-448, 1963) and Finegood et al. (Finegood, D., R. Bergman, and M. Vranic. Diabetes 36: 914-924, 1987) yielded acceptable approximations of predicted rates of glutamine plasma appearance with deviations from predicted rates from 0.2 to 1.6% (Finegood et al.) and from 0.1 to 8.2% (DeBodo et al.). Use of a 0.75 pool fraction most closely approximated predicted rates.

Regulation of gluconeogenesis by glutamine in normal postabsorptive humans.

There is evidence that glutamine may act as a regulator of protein, free fatty acid, and glycogen metabolism. To test the hypothesis that glutamine may act as a physiological regulator of gluconeogenesis, we infused 16 normal postabsorptive volunteers with glutamine at a rate (11.4 micromol kg(-1) x min(-1)) estimated to approximate its appearance in plasma after a protein meal and assessed changes in production of glucose from glutamine, systemic glucose appearance and disposal, and uptake and release of glucose, glutamine, and alanine by forearm skeletal muscle. Although infusion of glutamine increased plasma glutamine concentration and turnover only threefold (from 0.63 +/- 0.03 to 1.95 +/- 0.10 mmol/l and from 5.43 +/- 0.24 to 14.85 +/- 0.66 micromol x kg(-1) x min(-1), respectively; P < 0.001), formation of glucose from glutamine increased sevenfold from 0.55 +/- 0.03 to 3.74 +/- 0.28 micromol x kg(-1) x min(-1) (P < 0.001). Formation of glucose from alanine was also stimulated (0.52 +/- 0.05 vs. 0.75 +/- 0.04 micromol x kg(-1) x min(-1); P < 0.001) in the absence of a change in plasma alanine concentration. Furthermore, glutamine infusion decreased its own de novo synthesis (4.55 +/- 0.22 vs. 2.81 +/- 0.62 micromol x kg(-1) x min(-1);P < 0.02) while increasing that of alanine (2.82 +/- 0.32 vs. 3.56 +/- 0.32 micromol x kg(-1) x min(-1); P < 0.002). Systemic glucose appearance, systemic glucose disposal, and forearm balance of glucose and alanine were not altered. Because the stimulatory effects of glutamine on gluconeogenesis occurred in the absence of changes in plasma insulin and glucagon levels, these results provide evidence that, in humans, glutamine may act both as a substrate and as a regulator of gluconeogenesis as well as a modulator of its own metabolism.

Effect of age on muscle hypertrophy induced by resistance training.

BACKGROUND: Previous research has shown that resistance training induces muscle hypertrophy in older subjects, but has not clarified whether the degree of hypertrophy is affected by age. The present study was done to test the hypothesis that men and women over 60 years old have a smaller hypertrophic response to resistance training than young adults. METHODS: Cross-sectional areas (CSA) of muscle in the thigh and upper arm were determined before and after 3 months of progressive resistance training by magnetic resonance imaging (MRI) in 9 young (22-31 yr, 5 male and 4 female) and 8 old (62-72 yr, 4 male and 4 female) subjects. Strength was determined by 3-repetition-maximum (3RM) testing. The amount of weight lifted during the training program was proportional to baseline strength. RESULTS: Mean pretraining 3RM strength, per cm2 CSA, was less in the older group for all muscle groups examined (16 +/- 6% for elbow flexors, p < .02; 40 +/- 7% for knee flexors, p < .001; 19 +/- 9% for knee extensors, p < .05). Mean training-induced increases in muscle CSA were less in the older group for elbow flexors (22 +/- 4% in young, 9 +/- 4% in old, p < .05) and knee flexors (8 +/- 2% in young, 1 +/- 2% in old, p < .01), but not for knee extensors (4 +/- 1% in young, 6 +/- 2% in old). Mean training-induced increases in specific tension (ratio of 3RM strength to CSA) were similar in young and old groups for elbow flexors (21 +/- 5% in young, 19 +/- 5% in old) and knee extensors (38 +/- 6% in young, 32 +/- 14% in old), but were greater in the older group for knee flexors (28 +/- 5% in young, 64 +/- 13% in old, p < .02). CONCLUSIONS: Aging can attenuate the hypertrophic response of muscle groups to resistance training, when the training load is proportional to baseline strength. However, aging does not impair training-induced increases in specific tension.

Growth hormone increases muscle mass and strength but does not rejuvenate myofibrillar protein synthesis in healthy subjects over 60 years old.

The rate of synthesis of myofibrillar proteins is slower in muscle of healthy subjects over 60 yr old than it is in young adults. Previous research suggests that reduced activity of the GH/insulin-like growth factor-I system could be a determinant of this slowing of protein synthesis. To test the hypothesis that GH could rejuvenate the rate of myofibrillar protein synthesis, we studied healthy subjects over 60 yr old, after a single injection (0.03 mg/kg.sc) of recombinant human GH (n = 6 males/2 females) or placebo (n = 6 males/2 females), or after 3 months of either GH (0.03 mg/kg, sc, 3 x /week, n = 5 males) or placebo (n = 5 males) treatment. Myofibrillar protein synthesis and whole-body protein metabolism were evaluated with the tracer L-[1-13C]leucine. GH reduced whole-body leucine oxidation by 36% (P < 0.01) in the single injection study. There was no effect of GH on whole-body protein breakdown or synthesis, or on myofibrillar protein synthesis in the quadriceps, either acutely or after 3 months of treatment. GH treatment for 3 months increased lean body mass (3.3 +/- 0.7 kg, P < 0.01, as evaluated by 40K counting), muscle mass (3.3 +/- 1.1 kg, P < 0.02, as evaluated by urinary creatinine excretion), and thigh strength (14 +/- 5%, P < 0.05, as evaluated by isokinetic dynamometry). We conclude that GH can increase muscle mass and strength in healthy men over 60 yr old, but does not restore a youthful rate of myofibrillar protein synthesis.

Human IgG production in vivo: determination of synthetic rate by nonradioactive tracer incorporation.

Exposure of Ab-secreting cells to selected hormones, cytokines, and drugs alters the rate of Ig production in vitro. Whether these effects are important clinically is unknown because there are no safe, reproducible, and appropriate techniques for measuring Ig synthesis in vivo. We developed a stable isotope tracer method to measure IgG secretion into plasma. L-[1-13C]Leucine was given as a priming dose followed by a constant i.v. infusion over 8 h. Tracer accrual in IgG, determined by mass spectrometry, was linear from 2 to 8 h of the infusion. The normal rate of IgG synthesis into plasma assessed in 21 healthy subjects was 860 +/- 310 mg/day (mean +/- SD). The synthetic rate measurements were remarkably reproducible (coefficient of variation = 10.5 %). Simultaneous analysis of leucine kinetics allows, for the first time, Ig secretion to be studied in the context of whole body protein economy. IgG secretion into plasma accounts for 0.3% of whole body protein synthesis. Experimental support for a key metabolic assumption, that tracer enrichment in plasma and that at the site of IgG synthesis were similar, came from a comparison of synthetic rates derived from low dose and high dose tracer infusions. Measurement of Ig secretion by tracer incorporation is rapid and reproducible. In contrast to older methods that rely on radioisotope disappearance, the tracer incorporation method is safe for serial measurements in an individual and will be useful for quantitative studies of treatment effects and immune regulation in vivo.

Glutamine and alanine metabolism in NIDDM.

Gluconeogenesis is increased in NIDDM. We therefore examined the metabolism of glutamine and alanine, the most important gluconeogenic amino acids, in 14 postabsorptive NIDDM subjects and 18 nondiabetic volunteers using a combination of isotopic ([6-3H]glucose (20 microCi, 0.2 microCi/min), [U-14C]glutamine (20 microCi, 0.2 microCi/min), [3-13C]alanine (99% 13C, 2 mmol, 20 micromol/min), [ring-2H5]phenylalanine (99% 2H, 2 micromol/kg, 0.03 micromol x kg(-1) x min(-1)), and limb balance techniques. Alanine turnover (4.54 +/- 0.24 vs. 5.64 +/- 0.33 micromol x kg(-1) x min(-1)), de novo synthesis (3.00 +/- 0.25 vs. 4.01 +/- 0.33 micromol x kg(-1) x min(-1)), and conversion to glucose (1.02 +/- 0.09 vs. 1.56 +/- 0.17 micromol x kg(-1) x min(-1)) were increased in NIDDM subjects (all P < 0.01), while its forearm release (0.45 +/- 0.04 vs. 0.39 +/- 0.04 micromol x kg(-1) x min(-1)) was unaltered. Although glutamine turnover (4.81 +/- 0.23 vs. 4.40 +/- 0.31 micromol x kg(-1) x min(-1)) was unaltered in NIDDM, its conversion to glucose (0.57 +/- 0.04 vs. 1.08 +/- 0.10 micromol x kg(-1) x min(-1)) and to alanine (0.10 +/- 0.01 vs. 0.34 +/- 0.04 micromol x kg(-1) x min(-1)) (both P = 0.001) was increased while its oxidation (2.84 +/- 0.27 vs. 1.84 +/- 0.15 micromol x kg(-1) x min(-1), P = 0.03) and forearm release (0.77 +/- 0.05 vs. 0.62 +/- 0.09 micromol x kg(-1) x min(-1), P < 0.008) were both reduced. Our results thus demonstrate that there are substantial alterations of glutamine and alanine metabolism in NIDDM. Conversion of both amino acids to glucose and the proportion of their turnover used for gluconeogenesis are increased; release of both amino acids from tissues other than skeletal muscle seems to be increased. Finally, the reduction in glutamine oxidation, possibly the result of competition with glucose and free fatty acids as fuels, makes more glutamine available for gluconeogenesis without a change in its turnover.

Polyadenylated RNA, actin mRNA, and myosin heavy chain mRNA in young and old human skeletal muscle.

The myofibrillar protein synthesis rate in old human skeletal muscle is slower than that in young adult muscle. To examine whether this difference in protein synthesis rate is explained by reduced availability of the mRNAs that encode the most abundant myofibrillar proteins, we determined relative hybridization signals from probes for actin mRNA, myosin heavy chain mRNA, and total polyadenylated RNA in vastus lateralis muscle biopsies taken from young (22- to 31-yr-old) and old (61- to 74-yr-old) human subjects. The mean fractional rate of myofibrillar synthesis was 38% slower in the older muscles, as determined by incorporation of a stable isotope tracer. Total actin and myosin heavy chain mRNAs, and polyadenylated RNA, were determined using slot-blot assays. Isoform-specific determinations of alpha-actin mRNA, type I myosin heavy chain mRNA, and type IIa myosin heavy chain mRNA were done with ribonuclease protection assays. Hybridization signals were expressed relative to tissue DNA content. There was no difference between age groups in total polyadenylated RNA or in any of the specific mRNAs. We conclude that the slower myofibrillar synthesis rate in older muscle is not caused by reduced mRNA availability.

Utility of creatinine excretion in body-composition studies of healthy men and women older than 60 y.

This study evaluated whether aging alters the usefulness of creatinine excretion as an index of lean body mass (LBM) or muscle mass in healthy men and women. Creatinine excretion was determined while 24 young (15 men and 9 women aged < 32 y) and 38 older (23 men and 15 women aged > 60 y) healthy volunteers stayed at a clinical research center for 3 d. Total LBM was determined by 40K counting in all subjects, and cross-sectional areas of upper arm and thigh muscles were determined in some subjects (10 young, 20 old) by magnetic resonance imaging. The slopes and intercepts of the regression equations relating LBM to average daily creatinine excretion were the same in the younger and older groups, and the precision with which LBM could be determined from creatinine excretion was not diminished by aging. Creatinine excretion was closely correlated with cross-sectional areas of upper arm (r = 0.85) and thigh (r = 0.88) muscles, and the slopes and intercepts of the regression equations were not significantly affected by age. The older group had a mean LBM smaller than that of the younger group, which was accounted for entirely by a smaller muscle mass. Mean resting metabolic rate (RMR), whole-body protein turnover rate, and thigh strength were lower in the older group than in the young group. Adjustment of RMR and protein turnover for creatinine excretion or LBM eliminated the age effects. Strength was lower in the older group even after adjustment for creatinine excretion or LBM. This study indicates that creatinine excretion is useful for evaluating body composition in both young and old subjects.