Search PubMed⌕ Search

Biomedical subjects

J Wernerman

Publications and source records attributed to J Wernerman.

At least 73 records · Page 4Linked to original sources
PubMed1997

Longitudinal changes of biochemical parameters in muscle during critical illness.

The study was undertaken to characterize the time course of biochemical parameters in skeletal muscle during critical illness to gain information for the design of a suitable protocol for interventional studies using metabolic or nutritional manipulation. Critically ill patients in our intensive care unit ([ICU] N = 9) were investigated on two separate sampling occasions with percutaneous muscle biopsies for determination of protein, nucleic acids, free amino acids, energy-rich phosphates, fat, water, and electrolytes. The first biopsy specimen was taken 3 to 11 days after admission and the second biopsy specimen 3 to 7 days later. Protein concentration, expressed as alkali-soluble protein (ASP)/DNA, decreased by 12% (P < .02) between the two biopsies. The total free amino acid content was only 50% of normal, but remained unaltered over time. In particular, the concentration of glutamine remained low, approximately 25% of normal. In contrast, branched-chain amino acid (BCAA) increased by 25% (P < .05) and phenylalanine by 55% (P < .05) between biopsies. The fat content related to fat-free solid (FFS) increased by 130% (P < .001) between the two biopsies. Muscle water did not change during the study period. The extracellular portion was double the normal value when related to FFS. Intracellular water, on the other hand, was outside the 95% confidence interval for normal values in the second biopsy. The concentrations of adenosine triphosphate (ATP), creatine, phosphocreatine, and the phosphorylated fraction of total creatine remained at the same level between the two biopsies. We conclude that in critically ill patients, there is a decrease in protein content over time and increases in BCAA, phenylalanine, and fat content, while the low glutamine level and high extracellular water content remain unaltered. The temporal alterations were well characterized after a 5-day study period.

Adult↗
PubMed1997

Skeletal muscle glutathione is depleted in critically ill patients.

OBJECTIVE: To investigate the concentrations of reduced and total glutathione in relation to the muscle free amino acid pattern in critically ill patients and matched healthy controls. DESIGN: Prospective case control. SETTING: University hospital intensive care unit (ICU). PATIENTS: Eleven critically ill patients in the intensive care unit were studied after a stay of at least 4 days. Eleven age- and gender-matched metabolically healthy patients undergoing elective surgical procedures served as controls. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Reduced and total glutathione concentrations were determined in skeletal muscle, in plasma, and in whole blood, together with muscle free amino acid concentrations. In the ICU group, reduced and total glutathione values were 57% and 62%, respectively, of the values seen in the control group (p < .001). In addition, a decreased ratio between reduced and total glutathione compared with the controls was seen (0.80 as compared with 0.91, p < .001). The glutamine concentration in skeletal muscle in the ICU group was 72% lower compared with that value seen in healthy controls (p < .001). Correlations were found between the concentrations of glutamine and the total muscle glutathione (r2 = .46, p < .001), as well as between glutamine and the ratio of reduced and total glutathione (r2 = .45, p < .001) in skeletal muscle, suggesting that the redox status of glutathione and the glutamine status of the tissue are related. CONCLUSIONS: Critical illness is associated with alterations in muscle glutathione metabolism. The muscle-reduced glutathione concentrations decrease and, in addition, the ratio between reduced and total glutathione decreases, indicating a situation of oxidative stress in this tissue. This decrease may impair the defense of muscle against oxygen free radicals and influence amino acid transport, thus contributing to the loss of balance between protein synthesis and protein degradation that is characteristic of protein catabolism.

Adult↗
PubMed1997

Measurement of human growth hormone receptor messenger ribonucleic acid by a quantitative polymerase chain reaction-based assay: demonstration of reduced expression after elective surgery.

Studies of GH receptor (GHR) gene expression in human tissues have been hampered by the limited amount of tissue available for analysis and the low sensitivity of conventional methods. We have developed a quantitative reverse transcriptase-PCR assay for measurement of GHR messenger ribonucleic acid levels in small human tissue biopsies. To compensate for sample to sample variation, an internal RNA standard, which differs from the wild-type GHR transcript by only a few nucleotides, was reverse transcribed and amplified together with the GHR transcripts. PCR was carried out using one biotinylated primer to permit the purification of single stranded PCR products on streptavidin-coated microtiter plates. The ratio between the wild-type and mutated transcripts was determined by two separate minisequence reactions in which a primer, annealed immediately 3' of a variable nucleotide, was extended by a single 3H-labeled nucleotide, complementary to either the wild-type or mutated sequence. The assay range was 0.125-8 x 10(5) transcripts/sample, the mean intraassay coefficient of variation was 8.7%, and the lower limit of detection was 0.125 x 10(5) transcripts/sample. GHR messenger ribonucleic acid levels were detectable in small amounts (10-100 ng) of total RNA extracted from adipose tissue, skeletal muscle, and liver. The GHR gene expression in liver was approximately 10-fold higher than that in skeletal muscle, whereas intermediate levels were found in adipose tissue. In nine patients undergoing elective abdominal surgery, GHR gene expression in skeletal muscle was reduced on day 3 after surgery compared to the baseline level. The decrease in GHR gene expression was accompanied by a decrease in skeletal muscle glutamine. This suggests that the postoperative protein catabolism may be caused at least partly by acquired GH insensitivity due to reduced expression of the GHR gene.

Abdomen↗
PubMed1997

Correction of acidosis in dialysis patients increases branched-chain and total essential amino acid levels in muscle.

Earlier studies have shown increased oxidation of the branched-chain amino acids (BCAA), valine, isoleucine and leucine, in experimental acidosis and low levels of valine in the muscle of acidotic HD patients. Using HPLC, free amino acids in plasma and muscle were studied before and after correction of acidosis in 9 HD patients over 6 months by dialysis with a high bicarbonate solution. The predialysis standard bicarbonate concentration in blood increased from 20.6 +/- 1.3 mmol/l (mean +/- SD) before correction of acidosis to 25.9 +/- 1.8 mmol/l after correction. Correction of acidosis resulted in a significant increase in the i.c. concentrations of valine, isoleucine and leucine by 48%, 28% and 32%, as well as for the sum of BCAA and EAA, from a level lower than controls. The intra- and extracellular gradient increased for several amino acids and for the sum of EAA and BCAA, suggesting an increased influx or reduced efflux of amino acids across the cell membrane. Anthropometric data and the levels of S-albumin and transferrin did not change after correction of acidosis. The increases in the i.c. concentrations of BCAA after correction of acidosis suggest that the catabolism of these amino acids had been reduced. The effects of correction of acidosis on the concentrations of essential amino acids could be beneficial since low concentrations in muscle may reduce protein synthesis.

Acidosis↗
PubMed1996

Effect of transcutaneous electrical muscle stimulation on postoperative muscle mass and protein synthesis.

In an experimental study, 13 patients undergoing major elective abdominal surgery were given postoperative transcutaneous electrical muscle stimulation (TEMS) to the quadriceps femoris muscle on one leg; the opposite leg served as control. Changes in cross-sectional area (CSA) and muscle protein synthesis were assessed by computed tomography and ribosome analysis of percutaneous muscle biopsies before surgery and on the sixth postoperative day. The percentage of polyribosomes in the ribosome suspension decreased significantly (P < 0.03) after operation in control legs, but not in stimulated legs (P > 0.16). The total concentration of ribosomes decreased significantly in legs treated with TEMS (P < 0.03) but not in control legs (P > 0.16). CSA decreased significantly in both legs. The decrease in polyribosomes and CSA after operation was significantly less in stimulated legs than in controls (P < 0.05). TEMS may be a simple and effective method for improving muscle protein synthesis and muscle mass after abdominal surgery and should be evaluated in other catabolic states with muscle wasting.

Abdomen↗
PubMed1996

Protein synthesis rates of skeletal muscle, lymphocytes, and albumin with stress hormone infusion in healthy man.

The rate of protein synthesis was assessed in muscle, lymphocytes, and albumin in healthy volunteers administered an infusion of 6.0 micrograms cortisol +3.0 ng glucagon +0.5 nmol epinephrine min-1.kg-1. Protein synthesis in muscle tissue was not sensitive to the immediate effects of hormone infusion, but decreased significantly by 18 hours after the infusion had ceased (1.77% +/- 0.12% per day v 1.29% +/- 0.10%, P < .05). The rate of protein synthesis in lymphocytes was acutely sensitive to the effect of the hormone infusion, decreasing from 7.15% +/- 1.02% per day to 2.47% +/- 0.5% (P < .05). However, measurements made 18 hours after the end of the hormone infusion indicated that lymphocyte protein synthesis returned to the preinfusion rates. The rate of albumin synthesis was unaltered during infusion of the stress hormones, but was significantly increased when measured 18 hours after ending the hormone infusion (6.84% +/- 0.43% per day v 7.99% +/- 0.45%, P < .05). Thus, tissues respond differently to stress hormone infusion, demonstrating the importance of studying multiple organ systems when assessing the regulation of protein metabolism.

Adult↗
PubMed1996

Decrease in muscle glutamine, ribosomes, and the nitrogen losses are similar after laparoscopic compared with open cholecystectomy during the immediate postoperative period.

BACKGROUND: The purpose of the study was to compare the postoperative muscle amino acid pattern, the ribosome concentration and size distribution, and postoperative nitrogen balance in patients who underwent either laparoscopic or open cholecystectomy. METHODS: Patients who underwent cholecystectomy by means of either laparoscopy (n=8;LAP) or laparotomy (n=8;OPEN) were studied. The concentrations of amino acids, ribosomes, and polyribosomes, reflecting protein synthesis, were determined in skeletal muscle tissue before operation and on postoperative day 2. The cumulated nitrogen balance was determined. RESULTS. Decreases in muscle glutamine (26.7% +/- 8.4% in the LAP group and 30.3% and +/- 4.5% in the OPEN group) and in polyribosomes (28.7% +/- 6.5% in the LAP group and 23.6% +/- 8.5% in the OPEN group) were observed without differences between the groups (mean +/- SEM). The nitrogen losses were similar in both groups (15.2 +/-1.6 gm in the LAP group and 15.5 +/- 1.2 gm in the OPEN group). CONCLUSION: A stress++ response with effects on amino acid and protein metabolism in muscle in present also after laparoscopic cholecystectomy. On postoperative day 2 this response is of similar magnitude after both the laparoscopic and the open procedures.

Adult↗
PubMed1996

Effect of a short-term infusion of glutamine on muscle protein metabolism postoperatively.

The acute effect of a short-term postoperative infusion of glucose supplemented with glutamine (0.285 g/kg body weight), on muscle protein metabolism, was studied by analyses of free amino acid concentrations and determinations of protein synthesis. A glutamine-glucose infusion was given for 5.5 h to 6 patients 2-3 days after elective surgery for colon cancer. The free glutamine concentration was 5.72 +/- 0.96 mmol/kg wet weight (ww) before and 6.14 +/- 1.10 mmol/kg ww 4 h after the glutamine infusion. The rate of protein synthesis was 1.26 +/- 0.15%/24 h before the infusion and 1.12 +/- 0.16%/24 h during its latter part. The percentage of polyribosomes was 42.2 +/- 3.4% before and 40.9 +/- 1.3% after the infusion. The results showed no difference in these biochemical parameters, indicating that a short-term infusion of glutamine given postoperatively is insufficient to affect protein metabolism in human skeletal muscle.

Journal Article↗
PubMed1996

Protein metabolism in critical illness.

In summary, protein metabolism of critically ill patients is a field open to new investigations that will help us to understand better the mechanism behind 'autocannibalism', which is still today associated with mortality. Although the underlying disease is the major determinant of mortality, nutritional depletion will add morbidity, an addition that grows over time in the ICU. With conventional treatment the velocity of the catabolic process can at best be slowed down and the patient be bought time for other types of treatment to work. New forms of specific nutrition and adjuvant therapies may give us tools to prevent muscle depletion, without endangering the supply of essential substrates to the tissues in the splanchnic area. Muscle is at present a limiting organ for the ICU patient in two respects. A depleted muscle can no longer provide enough substrates for the splanchnic organs to maintain intestinal integrity and to maintain a high immunocompetence. In addition, a depleted muscle will be restored back to normal only very slowly; in elderly patients restoration may not even occur at all. The effects of an attenuation of muscle depletion on rehabilitation time have yet to be evaluated. An understanding of protein metabolism may be the key to better patient care in the ICU in the future.

Critical Illness↗
PubMed1996

Determination of protein synthesis in lymphocytes in vivo after surgery.

1. The stimulation and depression of peripheral blood lymphocytes has previously been studied in vitro, showing an immune depression postoperatively; however, it is difficult to interpret these in vitro findings. Therefore, an in vivo technique has been established for determination of the fractional protein synthesis rate, as an index of metabolic activity in human peripheral blood lymphocytes, by using a stable isotope technique. 2. The rate of protein synthesis was calculated from the increase in enrichment of L-[2H5]phenylalanine in protein of a mixed population of mononuclear leucocytes, isolated by density gradient, after an intravenous flooding dose of L-[2H5]phenylalanine. A linear time course of isotopic incorporation into the cells was demonstrated. 3. The fractional rate of protein synthesis of a mixed population of mononuclear leucocytes was studied in relation to surgical interventions and to potential modifiers of the response. The fractional synthesis rate increased 24 h after open and laparoscopic cholecystectomy (49 +/- 19% and 40 +/- 14% respectively, P < 0.02), irrespective of postoperative total parenteral nutrition or preoperative glucose infusion. In contrast to surgery, insulin did not stimulate protein synthesis in peripheral mononuclear leucocytes.

Adult↗
PubMed1996

Skeletal muscle glutathione after surgical trauma.

OBJECTIVE: The authors investigate the effect of surgical trauma on skeletal muscle concentrations of glutathione in patients undergoing selective abdominal surgery. SUMMARY BACKGROUND DATA: The posttraumatic state is accompanied by characteristic changes in the pattern of free amino acids and a decline of protein synthesis in human skeletal muscle. Glutathione has multiple metabolic functions that are involved in cellular homeostasis. It is unknown how surgical trauma affects the glutathione metabolism of skeletal muscle in surgical patients. METHODS: Eight patients undergoing elective abdominal surgery were investigated. Percutaneous muscle biopsies and blood samples were taken before operation and at 6, 24, and 48 hours after operation. The concentrations of glutathione were determined in muscle tissue, plasma, and whole blood, as well as the concentrations of the related amino acids in muscle and plasma. RESULTS: In skeletal muscle, the levels of both reduced and total glutathione decreased by 40% (p<0.01) at 24 hours and remained low at 48 hours after operation compared with the preoperative values. The glutathione concentration in plasma was 20% lower after operation compared with the concentration before operation (p<0.05). There were no changes at the whole blood levels of glutathione. Tissue glutamate and glutamine decreased significantly after operation (p<0.001), whereas intracellular cysteine and glycine remained unchanged. CONCLUSIONS: Skeletal muscle glutathione deficiency occurs after surgical trauma. This may lead to an increase in the susceptibility to intracellular oxidative injury.

Adult↗
PubMed1996

A descriptive study of skeletal muscle metabolism in critically ill patients: free amino acids, energy-rich phosphates, protein, nucleic acids, fat, water, and electrolytes.

OBJECTIVE: To characterize biochemical changes in skeletal muscle in critically ill patients. DESIGN: Survey of critically ill patients. SETTING: Intensive care unit (ICU) at a university hospital. PATIENTS: Critically ill patients (n = 20) subjected to trauma, surgical complications, and/or bacteremia who were treated in the ICU and showed no risk of bleeding complications were included. Reference groups of metabolically healthy volunteers and patients served as the control/reference groups. INTERVENTIONS: Percutaneous muscle biopsy was obtained from both patients and healthy volunteers. MEASUREMENTS AND MAIN RESULTS: Total free amino acids in skeletal muscle decreased 59% (p < .001) and skeletal muscle glutamine concentration decreased 72% (p < .001) in the critically ill patients. Basic amino acids decreased 49% (p < .001). Branch-chain amino acids increased 39% (p < .01), and aromatic amino acids increased 88% (p < .001) in the patients. Adenosine triphosphate (ATP) was reduced by 12% (p < .01). Total creatine concentration increased by 26% (p < .001) due to an 80% increase in free creatine (p < .001). The phosphorylated creatine fraction of total creatine decreased 22% (p < .001) in the patients. Alkali-soluble protein/DNA decreased 24% (p < .01) and fat free solid/DNA decreased 21% (P <.01) in patients sampled on or after ICU day 5 compared with the reference group. Muscle water increased 10% due to a doubling of the extracellular water fraction. CONCLUSIONS: Although critically ill patients are a very heterogeneous group from a clinical point of view, there is a remarkable homogeneity in many of the biochemical parameters regardless of the severity of illness and the length of the ICU admission. The three most consistent differences were the skeletal muscle low glutamine concentration, the decrease in protein content, and the increase in extracellular water in the patients.

APACHE↗
PubMed1996

The concentrations of free amino acids in human liver tissue obtained during laparoscopic surgery.

The concentrations of the free amino acids in individual tissues gives information concerning amino acid, energy and protein metabolism. In muscle and intestinal mucosa, different metabolic states are distinctly characterized by an altered free amino acid pattern. Furthermore, the patterns are quite different in individual tissues. So far, liver tissue has not been investigated systematically in this respect. The aims of this investigation were to establish a standardized sampling procedure for liver tissue during laparoscopic surgery and to characterize the free amino acid concentrations in human liver tissue. Aspartate was the most abundant amino acid in the liver, followed by taurine, glutamine, glutamate, glycine and alanine. These six, and most abundant, amino acids constitute 90% of the total hepatic amino acid concentration. In the future, liver tissue sampling during laparoscopic surgery may be used as a model for investigating the influence of nutrition and hormones on hepatic amino acid and protein metabolism in man.

Adult↗
PubMed1995

Determination of intracellular glutathione in human skeletal muscle by reversed-phase high-performance liquid chromatography.

A chromatographic method for the specific determination of cellular low molecular mass thiols has been applied to human muscle tissue. The method is based on the derivatisation of thiols using monobromobimane, which is a specific reagent for the sulphydryl group. The glutathione and cysteine bimane adducts were separated by reversed-phase HPLC, whilst quantitation of the cysteine and glutathione adducts was achieved by fluorescence spectroscopy. The method was found to yield a quantitative recovery of glutathione (ca. 96%), to be sensitive (down to 20 pmol glutathione/per injection) and reveal a low intra-individual coefficient of variation (C.V. < 5%) of the glutathione concentrations in human skeletal muscle. The concentrations of reduced and total glutathione were 1320 +/- 37 mumol/kg wet weight (mean +/- S.E.M.) and 1525 +/- 66 mumol/kg wet weight, respectively. The method was also applied to tissues from nine healthy volunteers to determine if fluctuations in glutathione level occurred over a 24-h period. No diurnal variation of glutathione level in human skeletal muscle was observed.

Adult↗