Biomedical subjects

B Quistorff

Publications and source records attributed to B Quistorff.

At least 37 records · Page 2Linked to original sources

PubMed1998

Sympathetic activation in exercise is not dependent on muscle acidosis. Direct evidence from studies in metabolic myopathies.

Muscle acidosis has been implicated as a major determinant of reflex sympathetic activation during exercise. To test this hypothesis we studied sympathetic exercise responses in metabolic myopathies in which muscle acidosis is impaired or augmented during exercise. As an index of reflex sympathetic activation to muscle, microneurographic measurements of muscle sympathetic nerve activity (MSNA) were obtained from the peroneal nerve. MSNA was measured during static handgrip exercise at 30% of maximal voluntary contraction force to exhaustion in patients in whom exercise-induced muscle acidosis is absent (seven myophosphorylase deficient patients; MD [McArdle's disease], and one patient with muscle phosphofructokinase deficiency [PFKD]), augmented (one patient with mitochondrial myopathy [MM]), or normal (five healthy controls). Muscle pH was monitored by 31P-magnetic resonance spectroscopy during handgrip exercise in the five control subjects, four MD patients, and the MM and PFKD patients. With handgrip to exhaustion, the increase in MSNA over baseline (bursts per minute [bpm] and total activity [%]) was not impaired in patients with MD (17+/-2 bpm, 124+/-42%) or PFKD (65 bpm, 307%), and was not enhanced in the MM patient (24 bpm, 131%) compared with controls (17+/-4 bpm, 115+/-17%). Post-handgrip ischemia studied in one McArdle patient, caused sustained elevation of MSNA above basal suggesting a chemoreflex activation of MSNA. Handgrip exercise elicited an enhanced drop in muscle pH of 0.51 U in the MM patient compared with the decrease in controls of 0.13+/-0.02 U. In contrast, muscle pH increased with exercise in MD by 0.12+/-0.05 U and in PFKD by 0.01 U. In conclusion, patients with glycogenolytic, glycolytic, and oxidative phosphorylation defects show normal muscle sympathetic nerve responses to static exercise. These findings indicate that muscle acidosis is not a prerequisite for sympathetic activation in exercise.

Acidosis↗

PubMed1998

Energy metabolism of the gastrocnemius and soleus muscles during isometric voluntary and electrically induced contractions in man.

1. Phosphocreatine (PCr) and intracellular pH detected by 31P NMR in the gastrocnemius and soleus muscles were evaluated in order to compare the anaerobic ATP costs of voluntary and electrically induced exercise. Continuous isometric contraction at 40% of maximum force and repeated isometric contractions at approximately 75% of maximum force (contraction plus relaxation period of 0.5 s plus 2 s) were studied. 2. Anaerobic ATP turnover in soleus and gastrocnemius muscles was slower during continuous voluntary contraction than during continuous electrically induced contraction (0.36 +/- 0.04 versus 0.63 +/- 0.05 mmol (kg wet wt)-1 s-1, P < 0.05, in soleus; 0.19 +/- 0.03 versus 1.04 +/- 0.04 mmol (kg wet wt)-1 s-1, P < 0.001, in gastrocnemius). 3. There was no significant difference in anaerobic ATP turnover between voluntary and electrically induced exercise when repeated brief contractions were performed (0.22 +/- 0.05 and 0.30 +/- 0.04 mmol (kg wet wt)-1 s-1, respectively, for the soleus muscle and 0.57 +/- 0.03 and 0.66 +/- 0.07 mmol (kg wet wt)-1 s-1, respectively, for the gastrocnemius muscle). 4. During continuous voluntary contraction, in contrast to continuous stimulated contraction, anaerobic ATP turnover was slower (P < 0.05) in the gastrocnemius than in the soleus muscle, which also showed a higher electromyogram amplitude (41.1 +/- 1.1% of maximum) than the medial gastrocnemius muscle (21.4 +/- 3.6% of maximum, P < 0.001). 5. Anaerobic ATP turnover was faster (P < 0.05) in the gastrocnemius than in the soleus muscle during brief voluntary and brief electrically induced contractions. 6. The results show that the anaerobic ATP costs were higher for electrically induced exercise than for voluntary exercise when continuous submaximal contraction was performed but not when brief high-intensity contractions were performed. The gastrocnemius muscle contributes to total force production relatively less than the soleus muscle during continuous voluntary plantar flexion at 40% of the maximum voluntary contraction.

Adenosine Triphosphate↗

PubMed1998

Effect of radiation therapy on small-cell lung cancer is reduced by ubiquinone intake.

The effect of oral ubiquinone (Q10) intake on the in vivo response of tumors to single dose radiotherapy was examined. The human small-cell lung cancer (SCLC) line CPH 054A, which is sensitive to relatively low doses of X-radiation, was grown as subcutaneous transplants in the flanks of nude nu/nu mice. When macroscopical growth was established, groups of mice received either 10, 20 or 40 mg/kg Q10 in 30 mL soy oil intragastrically daily on 4 consecutive days. Controls received either 30 mL of pure soy oil or nothing. Three h after the last dose half of the tumors in each group received a single radiation dose of 5 Gy, using a 300 kV therapeutic unit. The macroscopic growth pre- and posttreatment was analyzed according to a transformed Gompertz algorithm using the software program GROWTH. Treatment with Q10 or soy oil alone had no effect on tumor growth compared with untreated controls. Groups of tumors that received Q10 and radiotherapy had a significantly lower specific growth delay (SGD) than the radiotherapy-only groups. This effect was significant at 40 mg/kg and borderline at 20 mg/kg, whereas at 10 mg/kg no radioprotection was seen. We conclude that systemic Q10 reduces the response to single dose tumor irradiation inxenotransplanted human SCLC tumors.

Animals↗

PubMed1998

Effects of contraction duration on low-frequency fatigue in voluntary and electrically induced exercise of quadriceps muscle in humans.

The aims of this study were to investigate if low-frequency fatigue (LFF) dependent on the duration of repeated muscle contractions and to compare LFF in voluntary and electrically induced exercise. Male subjects performed three 9-min periods of repeated isometric knee extensions at 40% maximal voluntary contraction with contraction plus relaxation periods of 30 plus 60 s, 15 plus 30 s and 5 plus 10 s in protocols 1, 2 and 3, respectively. The same exercise protocols were repeated using feedback-controlled electrical stimulation at 40% maximal tetanic torque. Before and 15 min after each exercise period, knee extension torque at 1, 7, 10, 15, 20, 50 and 100 Hz was assessed. During voluntary exercise, electromyogram root mean square (EMGrms) of the vastus lateralis muscle was evaluated. The 20-Hz torque:100-Hz torque (20:100 Hz torque) ratio was reduced more after electrically induced than after voluntary exercise (P < 0.05). During electrically induced exercise, the decrease in 20:100 Hz torque ratio was gradually (P < 0.05) reduced as the individual contractions shortened. During voluntary exercise, the decrease in 20:100 Hz torque ratio and the increase in EMGrms were greater in protocol 1 (P < 0.01) than in protocols 2 and 3, which did not differ from each other. In conclusion, our results showed that LFF is dependent on the duration of individual muscle contractions during repetitive isometric exercise and that the electrically induced exercise produced a more pronounced LFF compared to voluntary exercise of submaximal intensity. It is suggested that compensatory recruitment of faster-contracting motor units is an additional factor affecting the severity of LFF during voluntary exercise.

Adult↗

PubMed1998

Motor unit activation patterns during concentric wrist flexion in humans with different muscle fibre composition.

Muscle activity was recorded from the flexor carpi radialis muscle during static and dynamic-concentric wrist flexion in six subjects, who had exhibited large differences in histochemically identified muscle fibre composition. Motor unit recruitment patterns were identified by sampling 310 motor units and counting firing rates in pulses per second (pps). During concentric wrist flexion at 30% of maximal exercise intensity the mean firing rate was 27 (SD 13) pps. This was around twice the value of 12 (SD 5) pps recorded during sustained static contraction at 30% of maximal voluntary contraction, despite a larger absolute force level during the static contraction. A similar pattern of higher firing rates during dynamic exercise was seen when concentric wrist flexion at 60% of maximal exercise intensity [30 (SD 14) pps] was compared with sustained static contraction at 60% of maximal voluntary contraction [19 (SD 8) pps]. The increase in dynamic exercise intensity was accomplished by recruitment of additional motor units rather than by increasing the firing rate as during static contractions. No difference in mean firing rates was found among subjects with different muscle fibre composition, who had previously exhibited marked differences in metabolic response during corresponding dynamic contractions. It was concluded that during submaximal dynamic contractions motor unit firing rate cannot be deduced from observations during static contractions and that muscle fibre composition may play a minor role.

Adult↗

PubMed1998

Muscle metabolism from near infrared spectroscopy during rhythmic handgrip in humans.

The rate of metabolism in forearm flexor muscles (MO2) was derived from near-infrared spectroscopy (NIRS-O2) during ischaemia at rest rhythmic handgrip at 15% and 30% of maximal voluntary contraction (MVC), post-exercise muscle ischaemia (PEMI), and recovery in seven subjects. The MO2 was compared with forearm oxygen uptake (VO2) [flow x (oxygen saturation in arnterial blood-oxygen saturation in venous blood, SaO2 - SvO2)], and with the 31P-magnetic resonance spectroscopy-determined ratio of inorganic phosphate to phosphocreatine (P(I):PCr). During ischaemia at rest, the fall in NIRS-O2 was more pronounced [76 (SEM 3) to 3 (SEM 1)%] than in SvO2 [71 (SEM 3) to 59 (SEM 2)%]. During the handgrip, NIRS-O2 was lower at 30% compared to 15% MVC [58 (SEM 3) v.s. 67 (SEM 3)%] while the SvO2 was similar [29 (SEM 3) v.s. 31 (SEM 4)%]. Accordingly, MO2 as well as P(I):PCr increased twofold, while VO2 increased only 30%. During PEMI after 15% and 30% MVC, NIRS-O2 fell to 9 (SEM 1)% and "0", but the use of oxygen by forearm muscles was not reflected in SvO2. During reperfusion after PEMI, the peak NIRS-O2 was lowest after intense exercise, while for SvO2 the reverse was seen. The discrepancies between NIRS-O2 and SvO2, and therefore between the estimates of the metabolic rate, would suggest significant limitations in sampling venous blood which is representative of the flexor muscle capillaries. In support of this contention, SvO2 and venous pH decreased during the first seconds of reperfusion after PEMI. To conclude, NIRS-O2 of forearm flexor muscles closely reflected the exercise intensity and the metabolic rate determined by magnetic resonance spectroscopy but not that rate derived from flow and the arterio-venous oxygen difference.

Adult↗

PubMed1998

Contribution of pH, diprotonated phosphate and potassium for the reflex increase in blood pressure during handgrip.

The relative importance of pH, diprotonated phosphate (H2PO4-) and potassium (K+) for the reflex increase in mean arterial pressure (MAP) during exercise was evaluated in seven subjects during rhythmic handgrip at 15 and 30% maximal voluntary contraction (MVC), followed by post-exercise muscle ischaemia (PEMI). During 15% MVC, MAP rose from 92 +/- 1 to 103 +/- 2 mmHg, [K+] from 4.1 +/- 0.1 to 5.1 +/- 0.1 mmol L-1, while the intracellular (7.00 +/- 0.01 to 6.80 +/- 0.06) and venous pH fell (7.39 +/- 0.01 to 7.30 +/- 0.01) (P < 0.05). The intracellular [H2PO4-] increased 8.4 +/- 2 mmol kg-1 and the venous [H2PO4-] from 0.14 +/- 0.01 to 0.16 +/- 0.01 mmol L-1 (P < 0.05). During PEMI, MAP remained elevated along with the intracellular [H2PO4-] as well as a low intracellular and venous pH. However, venous [K+] and [H2PO4-] returned to the level at rest. During 30% MVC handgrip, MAP rose to 130 +/- 3 mmHg, [K+] to 5.8 +/- 0.2 mmol L-1, the intracellular and extracellular [H2PO4-] by 20 +/- 5 mmol kg-1 and to 0.20 +/- 0.02 mmol L-1, respectively, while the intracellular (6.33 +/- 0.06) and venous pH fell (7.23 +/- 0.02) (P < 0.05). During post-exercise muscle ischaemia all variables remained close to the exercise levels. Analysis of each variable as a predictor of blood pressure indicated that only the intracellular pH and diprotonated phosphate were linked to the reflex elevation of blood pressure during handgrip.

Adult↗

PubMed1998

Regulation of glycogen accumulation in L6 myotubes cultured under optimized differentiation conditions.

The differentiation of the L6 myogenic cell line was enhanced by the addition of dexamethasone, retinoic acid, insulin-like growth factor I (IGF-I), and creatine. Spontaneous contractions appeared from day 10 or 11 and persisted to day 14 or 15. Glucose transport was increased by insulin (100 nM) and IGF-I (5 nM) by approximately 60%. The highest level of glycogen was measured in myotubes differentiated under the influence of a combination of 5 nM dexamethasone, 100 nM retinoic acid, 5 nM IGF-I, and 10 mM creatine with glucose as substrate. The glycogen accumulation rate was constant from 0 to 2 h of incubation and decreased gradually to zero at 4 h. From 0 to 0.5 h of the glycogen accumulation, the glycogen synthase a (GSa) activity was 30-35% of the total activity, with a subsequent gradual decline to 2.5% after 6 h. The glycogen phosphorylase a (GPha) activity was constant at approximately 80% from 0 to 0.5 h, increasing to approximately 100% after 6 h. The activity ratio of GSa to GPha decreased about sixfold without significant change in the rate of glycogen accumulation. This indicates that factors other than phosphorylation/dephosphorylation play a decisive role in the regulation of glycogen metabolism in L6 myotubes. Intracellular glucose (glucosei) and glucose 6-phosphate (G-6-P) may be such factors. The observed values of these parameters may in fact explain an activation of GSa (G-6-P) and an inhibition of GPha (glucosei).

Biological Transport↗

PubMed1997

Dipolar resonance frequency shifts in 1H MR spectra of skeletal muscle: confirmation in rats at 4.7 T in vivo and observation of changes postmortem.

Non-isotropic contributions to 1H MR spectra from human skeletal muscle in vivo have recently been observed in the 0- to 5-ppm region. One pair of peaks has been identified to be subject to dipolar couplings. The corresponding changes in resonance frequency are related to the orientation of muscle fibers with respect to the external magnetic field and are analogous to the behavior of small molecules dissolved in liquid crystals. Image-guided localized spectroscopy based on the STEAM method has been applied to verify these phenomena in rat skeletal muscle in vivo and to investigate the effect postmortem. Residual dipolar couplings and anisotropic contributions to 1H MR spectra of skeletal muscle have been confirmed in animals and at a higher field strength--albeit with a slightly different spectral pattern compared to the human study. The most prominent dipolar doublet due to creatine and/or phosphocreatine vanishes postmortem with a rate similar to the disappearance of phosphocreatine, and is no longer observable 2 h postmortem.

Animals↗

PubMed1997

Effects of oral supplementation of coenzyme Q10 on 31P-NMR detected skeletal muscle energy metabolism in middle-aged post-polio subjects and normal volunteers.

The effects of oral supplementation of 100 mg coenzyme Q10 (CoQ10) for 6 months on muscle energy metabolism during exercise and recovery were evaluated in middle-aged post-polio (n = 3) and healthy subjects (n = 4) by the use of phosphorus-31 nuclear magnetic resonance spectroscopy. The metabolic response to isometric plantar flexion at 60% of maximal voluntary contraction force (MVC) for 1.5 min was determined in gastrocnemius muscles before, after 3- (3MO) and 6-month (6MO) of CoQ10 supplementation. The MVC of plantar flexion was unchanged following CoQ10 supplementation. The resting Pi/PCr ratio in gastrocnemius muscles of all subjects decreased after 3MO- and 6MO-CoQ10 (P < 0.05). The post-polio individuals showed a progressive decrease in this ratio, while less pronounced changes were observed in the control subjects. Similarly, the post-polio individuals showed a lower Pi/PCr ratio at the end of 60% MVC in both 3MO- and 6MO-CoQ10, whereas no change in the ratio was observed in the control subjects. A less pronounced decrease in muscle pH was observed at the end of 60% MVC in both 3MO- and 6MO-CoQ10 in the post-polio individuals, but not in the control subjects. No systematic difference in end-exercise ATP was observed between the three phases in both groups. The half-time of recovery for PCr decreased in all subjects after 6MO-CoQ10 supplementation (P < 0.05). The results suggest that CoQ10 supplementation affects muscle energy metabolism in post-polio individuals to a greater extent than in control subjects. The mechanism for this effect is not clear, but may involve an effect of CoQ10 on peripheral circulation in the calf muscles, its action in mitochondrial oxidative phosphorylation and/or its antioxidant potential.

Administration, Oral↗

PubMed1996

Characterization of mitochondria from pig muscle: higher activity of exo-NADH oxidase in animals suffering from malignant hyperthermia.

Mitochondria were isolated from biopsies of the biceps femoris muscle of Danish landrace pigs. Three groups of animals were compared: (1) normal pigs; (2) pigs that were homozygous with respect to the gene Hal(n)/Hal(n) coding for the porcine malignant hyperthermia syndrome; and (3) heterozygote animals. A newly developed micro-method for preparation and assaying of small quantities of intact mitochondria was employed. With this technique mitochondria from biopsies weighing less than 100 mg were examined with respect to cytochrome content as well as phosphorylating and respiratory activities, including the nonphosphorylating exo-NADH oxidase activity. The mitochondria, prepared in a yield of 48%, showed high respiratory activities with tricarboxylic acid-cycle intermediates and pyruvate, and somewhat lower activity with palmitoyl-carnitine as substrate. The ATP synthase activity was about 1000 micromol ATP/min per g of protein and the maximal respiratory activity approx. 700 micromol of O2/min per g of protein. No differences among the three groups of animals were detected, except for the exo-NADH oxidase activities, which were 43, 78 and 107 micromol of O2/min per g of protein in the groups of normal, heterozygous and homozygous animals respectively. It is concluded that the exo-NADH oxidase activity may be a genetic manifestation of malignant hyperthermia and may play a significant role in the heat production characteristic of the syndrome.

Animals↗

PubMed1996

Assessment of dermal water by high-frequency ultrasound: comparative studies with nuclear magnetic resonance.

Although a principal constituent of human skin, cutaneous water is difficult to study, and little is known about water behaviour in physiological and pathological conditions of the skin. It has been proposed recently that changes in dermal echogenicity measured by high-frequency ultrasonography reflect changes in dermal water content. To validate skin ultrasonography for assessment of dermal water changes we have studied the relationship between dermal echogenicity and skin water content determined by nuclear magnetic resonance technique. Twenty MHz ultrasound scanning of the dorsal and ventral forearm skin was performed in 59 healthy volunteers (age 18-65) and dermal echogenicity was determined by counting low echogenic pixels (LEPs) in skin images. 1H magnetic resonance spectra were obtained from the same regions and the ratio of areas under the water- and fat-specific peaks (W/F) were calculated to measure a relative water content. Acute dermal oedema (histamine weal) was studied in the same way in 40 individuals. Baseline dermal echogenicity correlated significantly with W/F, both in the ventral (r = 0.47) and dorsal (r = 0.57) forearm. Intradermal application of histamine caused a development of intradermal oedema which could be visualized by nuclear magnetic resonance imaging. In a corresponding ultrasound image oedema was seen as a low-echogenic area. The proportional increases in LEPs and W/F after histamine application were correlated, but the elevation in LEPs was 25-48% (95% confidence intervals) higher than that for W/F. These results suggest that high-frequency ultrasonography is a sensitive method for assessment of changes in dermal hydration. This technique may find important applications in comparative and non-invasive evaluations of dermal water in physiological conditions and in skin pathologies associated with oedema formation.

Adolescent↗

PubMed1995

Growth inhibition in response to estrogen withdrawal and tamoxifen therapy of human breast cancer xenografts evaluated by in vivo 31P magnetic resonance spectroscopy, creatine kinase activity, and apoptotic index.

Estrogen withdrawal versus tamoxifen (TAM) treatment was compared in two human breast cancer xenografts, the estrogen-dependent ZR75-1 and its estrogen-independent subline ZR75/LCC-3. The following parameters were determined: tumor growth, NTP:P(i) by 31P magnetic resonance spectroscopy, apoptotic index, and creatine kinase (CK) activity. Tumors of each line were grown in ovariectomized nude mice during stimulation from a s.c. 17 beta-estradiol pellet. At a tumor size of approximately 350 mm3, the pellet was removed from one-half of the animals. The remaining one-half served as controls. In parallel experiments, injections of TAM were initiated instead of estrogen withdrawal. Estrogen withdrawal as well as TAM induced growth inhibition of ZR75-1 tumors, whereas ZR75/LCC-3 was resistant to both types of therapy. Growth inhibition of ZR75-1 by estrogen withdrawal, but not by TAM, was accompanied by an 80% increase of the NTP:P(i) ratio (P < 0.01) and a significantly decreased cytosolic CK activity (P < 0.01). No significant change in pH was observed. These changes seemed not to be related to changes in apoptotic index. None of the described changes occurred in ZR75/LCC-3. The present data indicate: (a) ZR75-1 and ZR75/LCC-3 xenografts respond differently to estrogen withdrawal and TAM with regard to growth inhibition, 31P magnetic resonance spectroscopy, and CK activity; (b) estrogen withdrawal, but not TAM, induced a decrease in the CK activity of estrogen-dependent tumor tissue, and (c) increased apoptosis did not explain the growth inhibition and the increase in NTP:P(i) induced by estrogen withdrawal. The results indicate other growth inhibitory mechanisms of TAM in addition to competitive inhibition of the estrogen receptor.

Animals↗

PubMed1995

Effect of estrogen withdrawal on energy-rich phosphates and prediction of estrogen dependence monitored by in vivo 31P magnetic resonance spectroscopy of four human breast cancer xenografts.

The effect of estrogen withdrawal on energy metabolism was studied in four human breast cancer xenografts: the estrogen-dependent MCF-7 and ZR75-1 and the estrogen-independent ZR75/LCC-3 and MDA-MB-231. The tumors were grown in ovariectomized nude mice with a s.c. implanted estrogen pellet. After Gompertzian growth was verified, the estrogen pellet was removed from half of the animals. In vivo 31P magnetic resonance spectroscopy of the tumors was performed 1 day before and on days 2, 6, and 14 after estrogen removal. Estrogen withdrawal induced a significant increase in the nucleoside triphosphate:Pi ratio in the two estrogen-dependent xenografts, whereas this ratio remained unchanged in the estrogen-independent tumors. In ZR75/LCC-3 tumors a slight decrease in nucleoside triphosphate:Pi was observed following onset of estrogen stimulation after initial growth without estrogen. Extracts of freeze-clamped tumors prepared 14 days after estrogen removal were analyzed for ATP and phosphocreatine content. Our findings suggest a correlation between estrogen withdrawal and the steady-state concentrations of ATP, phosphocreatine, and Pi in human breast cancer xenografts. Discrimination analysis of the pretherapeutic spectra enabled us to identify the tumor line and the estrogen dependence of the tumors in 80-90% of all cases.

Animals↗

PubMed1995

Ethanol inhibits acetate metabolism in rat hepatocytes.

The metabolism of acetate at concentrations of 1, 2.5, 5 and 10 mM was investigated in freshly isolated hepatocytes from 48 hr fasted, female rats in the absence and presence of 10 mM ethanol. The maximal capacity for acetate metabolism was 0.85 mumol/(10(8) cells.min). Ethanol caused a 20% decrease in the apparent Vmax for acetate metabolism and an increase in the apparent Km for acetate from 3.0 to 4.6 mM. At physiological concentration of acetate (approximately 1 mM) and in the absence of an inhibitory effect of ethanol, the capacity for acetate metabolism was 15-20% of the rate of acetate formation from ethanol and the inhibitory effect of ethanol further reduced it to 10-15%. The results thus explain the well-known but hitherto not understood fact that only a small fraction of acetate produced in the liver during ethanol oxidation is further metabolized by the liver, while the majority is exported for oxidation in other tissues. Finally, a new method for calculation of liver acetate uptake in the presence of ethanol is presented.

Acetates↗

PubMed1994

Kinetics of creatine uptake in the perfused mouse liver: a 31P-n.m.r. study of transgenic mice expressing creatine kinase (CKBB) in the liver.

Transport of creatine in the mouse liver has been investigated in vivo and in the perfused organ. Experiments were carried out with transgenic mice expressing creatine kinase in the liver (brain isoenzyme CKBB; EC 7.2.3.2.) [Koretsky, Brosnan, Chen, Chen and Van Dyke (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 3112-3116] and in the corresponding control mice. The animals were fed a regular chow with or without the addition of 10% creatine (w/w) for 5 days. The kinetics of creatine uptake was measured in the perfused liver by 31P-n.m.r. spectroscopy and biochemical analysis following infusion of creatine at concentrations ranging over 0-15 mM and at an extracellular pH of either 7.40 or 6.40. The results suggest that creatine is actively transported by a pH-dependent mechanism obeying a saturable Michaelis-Menten type of kinetics (Km = 0.80 +/- 0.18 and 5.12 +/- 2.40 mM; Vmax. = 0.57 +/- 0.04 and 1.72 +/- 0.32 mumol.g of liver-1.min-1 at pH 7.40 and 6.40 respectively). Creatine export was evaluated in the perfused liver preloaded with creatine and the results show that less than 2.5 and 5% of the total creatine pool is exported to the perfusate during 80 min of perfusion at pH 7.40 and 6.40 respectively. Taken together, these results seem to explain the observation that creatine accumulates in the mouse liver only when blood creatine is raised by creatine feeding.

Animals↗

PubMed1994

Hepatic metabolism during constant infusion of fructose; comparative studies with 31P-magnetic resonance spectroscopy in man and rats.

A protocol of constant infusion of fructose has been carried out both in human volunteers and in the perfused rat liver, aiming at a steady-state blood fructose concentration of 6-8 mM. Localized 31P-NMR spectroscopy and biochemical analyses were used to evaluate the metabolic changes. Comparison of the model experiment and the clinical study allowed an evaluation of this protocol as a clinically relevant assessment of the metabolic function of the liver. The time course of change, as well as the quasi steady-state levels reached during fructose infusion, for phosphomonoesters (PME), ATP and inorganic phosphate (Pi) provided the following results: During fructose infusion, ATP and Pi reached a steady-state level of 74.0 +/- 5.9 and 54.6 +/- 3.3% of control respectively, in the human volunteers. The corresponding data in the rat liver was 71.3 +/- 4.3 and 54.4 +/- 4.3%. Hepatic clearance of fructose was 0.53 and 0.52 ml.g liver-1.min-1 for volunteers and rats, respectively. The time course of intracellular metabolite recovery after fructose could be approximated by a first order kinetic. The rate constants for PME and ATP change were similar during fructose infusion and recovery, while after the discontinuation of fructose infusion, Pi increased with a rate constant significantly greater than during its fructose-induced depletion in human liver (P < 0.005). Thus, this relatively simple clinically applicable protocol seems to be verifiable in the well controlled perfused rat liver model, and it is argued that it may be useful in the clinical evaluation of the metabolic functional capacity of the human liver.

Adult↗

PubMed1994

Hepatic 5'-AMP-activated protein kinase: zonal distribution and relationship to acetyl-CoA carboxylase activity in varying nutritional states.

Acetyl-CoA carboxylase (ACC) activity in the liver varies markedly as a function of the nutritional state and is subject to complex regulation involving variable enzyme content, enzyme specific activity due to variable phosphorylation, and zonation within the hepatic lobule. A 5'-AMP-activated protein kinase (AMPK) has been identified as the major regulatory kinase active on ACC. Employing dual-digitonin pulse perfusion, the effect of varying nutrition on periportal and perivenous zonation of ACC and AMPK activity within the liver has been characterized. During the transition from fasting to refeeding with high-carbohydrate chow, total ACC activity is increased 11- to 17-fold. This induction of total ACC activity is accounted for by a 4.5- to 6-fold increase in the content of the two major ACC isoforms and by a 2.5-to 3-fold increase in enzyme specific activity (units per mg ACC). Despite a small perivenous preponderance of ACC protein, a gradient of activity to the periportal side, due to this increase in specific activity, is observed in fasted rats and during early refeeding. After 24-48 h of refeeding, maximal induction of both ACC protein and specific activity is observed with obliteration of this total activity gradient. 5'-AMP-activated protein kinase activity is maximal in the fasted rat and is zonated to the perivenous side. During refeeding, this activity is rapidly markedly diminished with abolition of this gradient, correlating inversely with the activation of ACC over a narrow range of kinase activity. Activities of casein kinase II, myelin basic protein kinase(s), and ribosomal S6 kinase(s) show no zonation. These data suggest that the zonal activity of the 5'-AMP-activated protein kinase is responsible, in part, for the intrahepatic gradient in ACC activity and that the regulation of this kinase is responsible for the variations in ACC-specific activity that occur during varying nutrition.

AMP-Activated Protein Kinases↗