Biomedical subjects

E Milne

Publications and source records attributed to E Milne.

At least 73 records · Page 4Linked to original sources

PubMed1993

Temporal responses of protein synthesis in human skeletal muscle to feeding.

In attempting to evaluate alterations in metabolic responses to dietary nutrients that occur in pathological conditions in man, it is first necessary to understand normal metabolic responses. The present study set out to determine the temporal responses of protein synthesis in the skeletal muscle of healthy subjects to the consumption of food. Sequential measurements of protein synthesis in quadriceps muscle were made in eight subjects by injection of 0.05 g L-[1-13C]leucine/kg body-weight. The rate of protein synthesis after an overnight fast (i.e. in the post-absorptive state) was 2.2% muscle protein. After 1 h of eating, protein synthesis was unaltered (2.2%/d), but after 10 h of consuming small hourly meals the rate had risen to 2.9%/d, with a variation in response among individuals. The response of muscle to 10 h of feeding was also investigated in subjects who underwent only one measurement each, either after 10 h of eating small meals or after the same time-period when no food was given. Protein synthesis rates were only slightly elevated in the group of fed individuals (2.3%/d, n 6) compared with the fasted group (2.1%/d, n 6). Taken together the two studies suggest that in healthy adults muscle protein synthesis does not respond quickly to the influx of dietary nutrients and that even after 10 h of feeding any stimulation of protein synthesis is small.

Adult↗

PubMed1993

An evaluation of the Northumberland Infectious Disease Report.

An evaluation of the quarterly Infectious Disease Report for Northumberland was undertaken by postal questionnaire. The response to the survey was encouraging, suggesting that this method of communicating local information is not only acceptable, but also effective and perceived as having a positive impact on practice.

Attitude of Health Personnel↗

PubMed1993

Muscle protein synthesis after operation: effects of intravenous nutrition.

OBJECTIVE: To assess the effect of an elective abdominal surgical operation (open cholecystectomy) on the rate of protein synthesis in skeletal muscle in humans. DESIGN: Prospective random control trial. SETTING: University hospital. SUBJECTS: 17 Metabolically healthy patients who were to undergo elective open cholecystectomy. INTERVENTIONS: Patients randomised to receive either saline alone (n = 8) or total parenteral nutrition (n = 9) for three days after operation. The rate of protein synthesis in muscle was calculated from the increase in enrichment of (1-13C) leucine in protein after a flooding dose of (1-13C) leucine. RESULTS: Median (quartiles) rate of protein synthesis had decreased on the third postoperative day in the saline group by 49% (from 2.42 [2.03, 2.54] to 1.24 [0.99, 1.63]) and in the group that had received total parenteral nutrition by 54% (from 1.96 [1.90, 2.07] to 0.91 [0.79, 1.06]) (p < 0.01). CONCLUSIONS: The trauma associated with open cholecystectomy reduced the rate of protein synthesis in skeletal muscle by half in three days, and conventional total parenteral nutrition had no effect on these changes.

Adult↗

PubMed1992

Turnover rates of different collagen types measured by isotope ratio mass spectrometry.

The rates of collagen turnover in different tissues have been estimated in growing rats previously exposed to gaseous 18O2. The abundance of the stable isotope was measured using isotope ratio mass spectrometry following combustion of isolated collagen-derived hydroxyproline. Using this method, problems of label reutilization associated with radiolabelling methods are avoided. In general the results confirm the slow turnover rates with half-lives of total collagen in skin, muscle and gut of 74, 45 and 244 d, respectively. The use of cyanogen bromide digests of whole tissues followed by isolation of collagen type-specific peptides has allowed the comparison of turnover rates of collagen types I and III, indicating that collagen type III is turned over more rapidly than type I.

Collagen↗

PubMed1992

Responses in tissue protein synthesis to sub- and supra-maintenance intake in young growing sheep: comparison of large-dose and continuous-infusion techniques.

In ten lambs (average live weight 33 kg), five offered 300 g/d (approximately 0.6 x maintenance; L) and five 900 g/d (1.8 x maintenance; H), tissue protein synthesis was measured by three procedures simultaneously. The techniques involved continuous infusion of [U-14C]phenylalanine and [1-13C]leucine over 7-8 h followed by a terminal large dose of [15N]phenylalanine during the last 30 or 60 min. Rates of protein synthesis were then calculated based on the free amino acid or oxo-acid isotopic activity in either arterial, iliac venous blood or tissue homogenate for the continuous-infusion studies, or on plasma or tissue homogenate for the large-dose procedure. For muscle (> 99%), and to a lesser extent skin (85-93%), effective flood conditions were achieved with the [15N]phenylalanine but were either not established or maintained for liver and tissues of the gastrointestinal tract (< 50%). The large dose of phenylalanine also caused changes in the concentration and isotopic activity of blood leucine and 4-methyl-2-oxo-pentanoate. Based on the assumption that the large-dose procedure yields the closest value for the true rate of protein synthesis (L 1.97%/d, H 2.85%/d) then, for muscle, only values based on the homogenate as precursor gave comparable results for both leucine (L 1.83%/d, H 3.01%/d) and phenylalanine (L 1.67%/d, H 2.71%/d) continuous infusion. The values based on the arterial or venous amino or oxo-acid were significantly less, more so at the lower intake. In contrast, for skin, a tissue dominated by export protein synthesis, values from the large-dose procedure (L 6.37%/d, H 10.98%/d) were similar to those derived with arterial or venous metabolites as precursor (L 5.23 and 6.93%/d, H 9.98 and 11.71%/d for leucine), but much less than those based on homogenate data. Based on the large-dose technique, protein synthesis increased with intake in muscle (P < 0.001), skin (P = 0.009) and liver (26.7 v. 30.5%/d; P = 0.029). The contributions of muscle and skin to total protein synthesis were approximately equal. The incremental efficiency of conversion for muscle of synthesized protein into deposition appeared to be similar to values reported for rodents.

Animals↗

PubMed1992

Effect of food intake on hind-limb and whole-body protein metabolism in young growing sheep: chronic studies based on arterio-venous techniques.

Whole-body protein synthesis, estimated by the irreversible loss rate procedure, and hind-leg protein metabolism determined by arterio-venous techniques were monitored in response to three nutritional conditions (approximately 0.6, 1.2 and 1.8 x energy maintenance (M)) in ten wether lambs (33 kg average live weight). In all lambs and treatments measurements were based on radiolabelled phenylalanine, but the terminal procedures (five at 0.6 x M and five at 1.8 x M) also included infusion of [1-13C]leucine; this permitted comparison of amino acids catabolized (leucine) and non-metabolized (phenylalanine) by the hind-limb tissues. Whole-body protein synthesis increased with intake and the relationship with energy expenditure was slightly lower than that reported previously for pigs and cattle. The efficiency of protein retention:protein synthesis did not exceed 0.25 between the two intake extremes. Effects of intake on amino acid oxidation were similar to those observed for cattle. Hind-limb protein synthesis also increased significantly (P < 0.001) in response to intake. Estimates of protein gain, from net uptake values, indicated that the tissues made a greater proportional contribution to total protein retention above M and to protein loss below M, emphasizing the role played by muscle tissue in providing mobile protein stores. The rates of protein synthesis calculated depended on the selection of precursor (blood) metabolite, but rates based on leucine always exceeded those based on phenylalanine when precursor from the same pool was selected. The incremental efficiency of protein retained:protein synthesis was apparently unity between 0.6 and 1.2 x M but 0.3 from 1.2 to 1.8 x M. Blood flow through the iliac artery was also proportional to intake. Leucine and oxo-acid catabolism to carbon dioxide increased with intake such that the metabolic fate of the amino acid was distributed in the proportion 2:1 between protein gain and oxidation. The rates of oxidation were only 1-3% the reported capacity of the rate-limiting dehydrogenase enzyme in muscle, but sufficient enzyme activity resides in the hind-limb adipose tissue to account for such catabolism.

Amino Acids↗

PubMed1992

Short-term starvation decreases skeletal muscle protein synthesis rate in man.

The rate of protein synthesis in skeletal muscle was determined in the post-absorptive state and after 3 days of starvation in healthy volunteers. The flooding dose technique employing intravenous injection of (1-13C)leucine (0.05 g kg-1) was used and incorporation of isotope into muscle protein was measured by taking percutaneous biopsies at 0 and 90 min. Blood samples were taken during the incorporation period for assessment of the enrichment of the free amino acid precursor of protein synthesis. The median (25,75 quartiles) rate of muscle protein synthesis after an overnight fast was 2.03 (2.00,2.23) % days-1 when the precursor enrichment was obtained by measurement of the plasma alpha-ketoisocaproate, taken to be representative of muscle free leucine. Repeat measurements in the same subjects after 3 days of total starvation showed a decrease to 1.82 (1.57,2.05) % days-1. Rates calculated on the basis of the plasma leucine as precursor were 5% lower at both times. An interindividual variation in response to starvation was observed, but the median decrease of 13% in the rate of protein synthesis was statistically significant (P less than 0.01).

Adult↗

PubMed1991

Stimulation of protein synthesis in human tumours by parenteral nutrition: evidence for modulation of tumour growth.

Eighteen patients with localized colorectal carcinoma were randomized to receive intravenous nutrition or to be fasted during the 24 h before surgery. Protein synthesis, an index of tumour growth, was then measured by the incorporation of [13C]leucine into tumour protein immediately before surgery. The mean (s.e.m.) rate of tumour protein synthesis in patients receiving nutrition (42.7(3.5) per cent per day) was 89 per cent higher than the rate in the fasted group (22.6(1.9) per cent per day) (P = 0.002). As tumours consist of a variety of different cell types, in vitro rates of protein synthesis were measured in malignant cells isolated from colorectal tumours and cultured with autologous serum obtained from the patient in either the fasted or the fed state. There was a mean increase of 81 per cent in protein synthesis when fed rather than fasted serum was used (P less than 0.02), indicating that the malignant cells themselves respond to nutrient supply. This increase in tumour protein synthesis provides the first evidence in vivo that the exogenous supply of nutrients can modulate the rate of growth of a human tumour.

Aged↗

PubMed1991

The effect of insulin suppression on postprandial nutrient metabolism: studies with infusion of somatostatin and insulin.

Utilization of fat, carbohydrate and protein before and after feeding was studied in six healthy subjects using simultaneous respiratory gas exchange measurement and [1-13C] leucine infusion. The role of insulin was investigated by repeating a control study with the addition of an infusion of somatostatin, a hormone which can suppress insulin release. Where near-complete insulin suppression was effected, subjects were studied on a third occasion with the further addition of exogenous insulin infusion. The normal switch on feeding from fat to carbohydrate as principal energy source was reproduced at insulin levels of only 17%-33% of control values, which were inadequate to prevent hyperglycaemia. At fed levels below 10%, a fat-predominant pattern persisted unless insulin was infused. Protein degradation was reduced and synthesis unaffected by feeding, regardless of insulin concentration. Leucine oxidation was dependent on its plasma concentration in the presence of circulating insulin. Thus insulin appears to be necessary for the normal switch to carbohydrate oxidation on feeding but not for postprandial changes in protein metabolism.

Adult↗

PubMed1990

Baseline measurements for stable isotope studies: an alternative to biopsy.

Studies of protein metabolism with stable isotopes require determination of the 'natural' isotopic enrichment in tissues. This has previously been determined by taking a pre-test muscle biopsy or by using the isotopic enrichment of a separate control group of subjects. In this study we have measured and compared the 'natural' 13C enrichment of leucine in plasma protein and muscle protein in 14 subjects. The mean enrichment of leucine (delta 13CPDB) in muscle protein, -26.627, was not significantly different from that in plasma protein, -27.152. The data indicate that the 13C enrichment of leucine in plasma protein reflects that of muscle protein and provides an attractive alternative to an additional muscle biopsy in studies of protein metabolism with stable isotopes.

Adult↗

PubMed1990

Muscle protein synthesis in response to testosterone administration in wether lambs.

A method has been developed based on stable isotopes and biopsy procedures which allows the large-dose procedure for measurement of protein synthesis to be applied in serial studies to farm species. Measurements of total nitrogen retention and protein synthesis in m. longissimus dorsi and m. vastus lateralis were made in five wether lambs (40-44 kg) infused intravenously, successively, with vehicle (10 d); testosterone (15 d; 9 mg/d); vehicle (15 d). N retention was improved by testosterone infusion (+2.9 g N/d; a 96% improvement total over control periods). Muscle protein synthesis was not significantly altered by exogenous hormone administration, nor were RNA:protein, RNA:DNA or protein:DNA. The implication of the developed procedure for dynamic studies in accessible tissues of large animals is discussed.

Animals↗

PubMed1990

Measurement of albumin synthesis in humans: a new approach employing stable isotopes.

A new method for measuring albumin synthesis in humans with stable isotopes is presented. This can readily be applied in most clinical conditions, even when albumin losses are occurring or when repeated assessment is required. After rapid intravenous injection of a large dose of [13C]leucine (57 mg/kg body wt, 19.4 atoms%), plasma samples were taken at intervals up to 90 min. The enrichment of free leucine in plasma measured by gas chromatography-mass spectrometry rose to a peak at 10 min and then fell slowly, whereas that in liver biopsies (from surgical patients) ranged from 101.5 to 80.5% of the plasma value between 10 and 90 min after injection. The fractional synthesis rate (FSR) was calculated by dividing the increase in enrichment of leucine in albumin, measured by gas isotope ratio mass spectrometry, by the area under the plasma free leucine enrichment vs. time curve after allowing for the period between synthesis of the protein and its secretion into the plasma. The FSR in healthy postabsorptive males was 7.2 +/- 1.3%/day, and the absolute synthesis rate was 157 +/- 39 mg.kg body wt-1.day-1. These rates are comparable to those obtained by other methods.

Adult↗

PubMed1990

Nutrient oxidation patterns and protein metabolism in lean and obese subjects.

The immediate metabolic response to eating has been compared in a group of grossly obese subjects (W/H2 = 45) with that in lean controls (W/H2 = 22). Dietary intake of energy for obese subjects was based on their estimated basal energy expenditure for ideal body weight (given at an hourly rate of 3 X BMR over a 4-h period). Lean subjects were measured twice: control 1 with the same intake of energy as the obese in terms of ideal body weight and control 2 with the same energy intake in relation to each subject's measured resting energy expenditure (2.2 X REE). The changes in energy expenditure and nutrient disposal with the onset of eating have been assessed by a method of combined respiratory gas analysis and intravenous infusion of 13C-labelled leucine. Leucine kinetics were used to quantitate rapid changes in protein oxidation and to assess protein synthesis and degradation. 1) Total energy expenditure was 20-30 per cent greater in obese subjects than lean subjects in fasting and feeding. Energy expenditure expressed per kg fat-free mass, from D2O dilution, was similar in obese and lean subjects in both fasting (5.8 v. 5.5 kJ/kg FFM/h) and feeding [6.7 v. 6.3 (Control 2) kJ/kg FFM/h]. 2) The onset of eating was associated with increased carbohydrate and protein oxidation with decreased fat oxidation in both lean and obese individuals. In obese subjects, however, both the decrease in fat oxidation and the increase in protein oxidation were significantly smaller (P less than 0.05) than the corresponding increments in lean subjects (Control 2). 3) The rate of protein synthesis was significantly (P less than 0.05) higher in obese subjects both in the fasting state (99 v. 84 mumols leucine/kg FFM/h) and in the fed state [94 v. 67 (Control 2) mumols leucine/kg FFM/h]. The rate of protein degradation was also higher in obese individuals in fasting (117 +/- 6 v. 106 +/- 4 mumol leucine/kg FFM/h) and feeding [65 +/- 4 v. 54 +/- 6 (Control 2) mumol leucine/kg FFM/h] though these differences are not statistically significant (P greater than 0.05). 4) The observed differences between obese and lean individuals in protein and energy metabolism in the fasted state and in the immediate response to eating do not support a hypothesis of greater metabolic efficiency in obesity.

Adult↗

PubMed1989

The role of degradation in the acute control of protein balance in adult man: failure of feeding to stimulate protein synthesis as assessed by L-[1-13C]leucin infusion.

The effect of feeding on whole-body protein turnover was measured in six healthy volunteers using the essential amino acid, L-[1-13C]leucine, as a tracer for protein metabolism. Varied lengths of periods of feeding and isotope infusion produced different apparent responses to feeding. When parameters of protein turnover were estimated from 8-hour infusions, the change from post-absorptive in the first four hours to mixed feeding during the final four hours was found to produce positive leucine balance by decreasing degradation from 89.5 +/- 5.0 to 31.7 +/- 7.3 mumol leucine/kg/h (P less than .001), with no apparent change in synthesis. By contrast, when tracer was infused for 24 hours with 12 hours of feeding followed by 12 hours of fasting, the estimate of protein synthesis during feeding was 35% higher than during fasting (P less than .01). However, when tracer infusion during the 12-hour feeding/12-hour fasting protocol was limited to the last four hours of each nutritional period, the estimates of fed and fasted protein synthesis showed no significant difference, 71.3 +/- 6.5 and 66.2 +/- 5.6 respectively, while the calculated rate of protein degradation was 43% lower during feeding (P less than .002). As relatively higher levels of enrichment in plasma leucine were detected in comparable nutritional states following longer infusions, the possibility of significant recycling of label was investigated. Residual tracer was still detectable in both breath and plasma 12 hours after cessation of a 12-hour tracer infusion, supporting the conclusion that significant errors in estimates of protein turnover due to recycling of label arise with prolonged infusions.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1989

Measurement of the rate of protein synthesis in muscle of postabsorptive young men by injection of a 'flooding dose' of [1-13C]leucine.

1. The 'flooding dose' technique for measuring the rate of protein synthesis in tissues in vivo involves the injection of a large amount of unlabelled amino acid together with the tracer to minimize differences in isotopic enrichment of the free amino acid in plasma and tissue compartments. This approach has been investigated in human muscle by taking biopsies from postabsorptive male volunteers given [1-13C]leucine. 2. Intravenous injection of 4 g of unlabelled leucine resulted in a rapid rise in free leucine concentration of seven- to eleven-fold in plasma and five-fold in muscle. Values were still elevated by two-fold after 2 h. 3. Five minutes after injection of [1-13C]leucine (0.05 g/kg) the isotopic enrichment of plasma leucine was 82% that of the injected material, falling to 44% at 120 min. The enrichment of free leucine in sequential muscle biopsies was close to that in plasma and almost identical to that for plasma alpha-ketoisocaproate. 4. The rate of protein synthesis was determined from the increase in leucine enrichment in protein of muscle biopsies taken before and 90 min after injection of [1-13C]leucine (0.05 g/kg; 19 or 39 atom% excess) and the average plasma alpha-ketoisocaproate enrichment over this period (taken to represent muscle free leucine). The mean rate of muscle protein synthesis in 10 subjects was 1.95 (SEM 0.12) %/day. Rates of protein synthesis calculated from plasma leucine as precursor enrichment were only 5% lower than those calculated from plasma alpha-ketoisocaproate.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1988

The applicability of evacuated serological tubes for the collection of breath for isotopic analysis of CO2 by isotope ratio mass spectrometry.

Stable isotope ratio mass spectrometry was used for the analysis of 13C and 18O in CO2 of human breath collected in evacuated serological tubes (Vacutainer and Venoject) and in gas sample bottles. There was a 1-2% difference between the delta 13 CPDB values obtained from breath CO2 collected in gas bottles and that collected in two batches of Vacutainer. delta 18OPDB differed between Vacutainers and gas bottles by as much as 14% and standard deviations within the two batches of Vacutainer were 0.7 and 1.8%. The poor reproducibility and accuracy for the delta 18OPDB values was caused by the presence of a contaminant which originated from the rubber septa of the Vacutainers. The results show that it is not possible to obtain delta 18OPDB values with a high degree of accuracy or precision for breath samples collected in Vacutainers. However, with selection of less contaminated or re-evacuated batches, delta 13CPDB analysis of breath CO2 in Vacutainers may provide acceptable accuracy and precision. delta 13CPDB and delta 18OPDB values measured in breath collected in non-sterile Venoject tubes were also significantly different from those obtained with gas bottles. However, the accuracy and precision of these determinations were considerably better than in sterilized Vacutainer tubes. As a result of these comparisons it is concluded that gas bottles are necessary for the full accuracy and precision of stable isotope ratio analysis mass spectrometers to be exploited in breath CO2 analysis. However, automated analysis of breath CO2 collected in non-sterile Venoject tubes will provide an accuracy and precision that is suitable for most biological studies.

Breath Tests↗

PubMed1987

The effect of indomethacin on the response of protein synthesis to feeding in rats and man.

Protein synthesis was measured in muscle of fed and fasted rats. Synthesis rates were 29% higher with feeding. Treatment with indomethacin before feeding completely blocked the stimulation of protein synthesis. Whole-body protein synthesis was measured in six healthy volunteers with [15N]glycine in fasted and fed states. Feeding increased synthesis rates by 50%. An oral dose of indomethacin 2 h before the measurement of protein synthesis had no effect in either the fasted or fed state.

Adult↗

PubMed1987

Rates of nutrient utilization in man measured by combined respiratory gas analysis and stable isotopic labelling: effect of food intake.

Rates of oxygen consumption and carbon dioxide production have been measured in healthy adults during 4 h of fasting followed by 4 h of hourly small meals. Both rates rose to new steady values during feeding, and the respiratory quotient (RQ) increased from 0.792 to 0.924. The RQ was consistent in repeat studies on any individual (coefficient of variation: 2.5 per cent), and differences between individuals were significant in the fasted but not the fed state. Simultaneous measurements were made of the rate of protein oxidation by primed constant infusion of (1-13C)leucine for 8 h. Rates were calculated from the enrichment of plasma alpha-ketoisocaproate and the production of 13CO2 in the breath, taking account of the incomplete recovery of 13CO2 and the changes in baseline enrichment resulting from natural 13C in the food. Leucine oxidation increased by 87 per cent during the feeding period. Rates of nutrient utilization were calculated from respiratory gas exchange and rates of protein oxidation. These showed that fat was predominant in the fasted state, contributing 61 per cent of total energy expenditure, compared with 27 per cent for carbohydrate and 11 per cent for protein. On feeding there was a switch to carbohydrate as the main fuel (62 per cent), with smaller contributions from fat (20 per cent) and protein (18 per cent). During feeding total utilization of each nutrient exceeded its intake from the diet, indicating storage in the body. Dietary carbohydrate was stored without conversion to fat. It is concluded that this method is useful for studying the control of nutrient utilization by food intake.

Adult↗