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Objective: Topical negative pressure, commonly used in wound therapy, has been shown to increase blood flow and stimulate angiogenesis in subcutaneous tissue and skeletal muscle. In wound therapy, intermittent negative pressure is often preferred to continuous negative pressure as tissue exposed to intermittent therapy shows twice as much granulation tissue formation than that exposed to continuous pressure after 2 weeks of therapy. The present study was designed to elucidate the differences in microvascular blood flow in the left anterior descending artery area between continuous and intermittent myocardial topical negative pressure of -50 mmHg., Methods: Six pigs underwent median sternotomy. Laser Doppler probes were inserted horizontally into the heart muscle in the left anterior descending artery area at depths of approximately 5-6 mm. Measurements of microvascular blood flow were performed in normal myocardium and ischemic myocardium during 20 min of countinuous and intermittent topical negative pressure at -50 mmHg., Results: Both continuous and intermittent topical negative pressure of -50 mmHg significantly increased microvascular blood flow in the underlying myocardium: from 56.2 +/- 13.1 perfusion units (PU) before to 132.8 +/- 7.4 PU during countinuous topical negative pressure application (P < 0.05) and from 75.8 +/- 12.1 PU before to 153.6 +/- 4.7 PU during intermittent topical negative pressure application (P < 0.05)., Conclusion: No statistically significant difference was found between microvascular blood flow during 20 min of continuous and intermittent topical negative pressure at -50 mmHg in this porcine model.

Objective: We previously reported the induction of stable immune tolerance following direct injection of retroviral vectors into preimmune fetal sheep. In the present studies, we conduct detailed analysis of the thymus of recipients of in utero gene transfer (IUGT) to delineate the mechanism of the observed immune tolerance and assess the impact of recipient age on this process., Materials and Methods: Fetal sheep at varying gestational ages received the MSCV-NeoR-RFP retroviral vector. The thymus was then collected from these animals at 27 to 30 days postinjection and analyzed for evidence of transduction of key immunoregulatory thymic cells., Results: Our results reveal that both thymic epithelial cells (TEC), crucial for presentation of self-antigen during T-cell thymic selection, and the cells comprising the Hassall's corpuscles, which can present antigen directly and also instruct dendritic cells to induce the formation of CD4(+)CD25(+) T-regulatory cells in the thymus, were only efficiently transduced if IUGT was performed early in gestation., Conclusions: Our findings thus demonstrate, for the first time, that early IUGT can potentially take advantage of multiple tolerogenic avenues in the fetus, transducing both TEC, which promote central tolerance, and Hassall's corpuscles, which induce formation of T regulatory cells that could act to maintain peripheral tolerance to the transgene products.

Background: We have previously shown that a myocardial topical negative pressure (TNP) of -50 mmHg significantly increases microvascular blood flow in the underlying myocardium in normal, ischemic, and reperfused porcine myocardium. The present study was designed to elucidate the effect of different TNP levels between -50 and -150 mmHg on microvascular flow in normal and ischemic myocardium., Materials and Methods: Seven pigs underwent median sternotomy. The microvascular blood flow in the myocardium was recorded, before and after the application of TNP, using laser Doppler velocimetry. Analyses were performed before left anterior descending artery (LAD) occlusion (normal myocardium) and after 20 min of LAD occlusion (ischemic myocardium)., Results: A TNP of -50 mmHg significantly increased microvascular blood flow in both normal (from 320.0+/-56.1 PU before TNP application to 435.7+/-65.5 PU after TNP application, P=.028) and ischemic myocardium (from 110.0+/-36.7 PU before TNP application to 194.3+/-56.2 PU after TNP application, P=.012). TNP between -75 and -150 mmHg showed no significant increase in microvascular blood flow in normal or ischemic myocardium., Conclusions: Of pressures between -50 and -150 mmHg, a TNP of -50 mmHg seems to be the most effective negative pressure concerning significant increase in microvascular blood flow in both normal and ischemic myocardium.

Background: Topical negative pressure (TNP), commonly used in wound therapy, has been shown to increase blood flow and stimulate angiogenesis in skeletal muscle. We have previously shown that a myocardial TNP of -50 mmHg significantly increases microvascular blood flow in the myocardium. When TPN is used in wound therapy (on skeletal and subcutaneous tissue) a zone of relative hypoperfusion is seen close to the wound edge. Hypoperfusion induced by TNP is thought to depend on tissue density, distance from the negative pressure source, and the amount negative pressure applied. When applying TNP to the myocardium, a significant, long-standing zone of hypoperfusion could theoretically cause ischemia, and negative effects on the myocardium. The current study was designed to elucidate whether hypoperfusion was produced during myocardial TNP., Methods: Six pigs underwent median sternotomy. Laser Doppler probes were inserted horizontally into the heart muscle in the LAD area, at depths of approximately, 1-2 mm. The microvascular blood flow was measured before and after the application of a TNP. Analyses were performed before left anterior descending artery (LAD) occlusion (normal myocardium) and after 20 minutes of LAD occlusion (ischemic myocardium)., Results: A TNP of -50 mmHg induced a significant increase in microvascular blood flow in normal myocardium (**p = 0.01), while -125 mmHg did not significantly alter the microvascular blood flow. In ischemic myocardium a TNP of -50 mmHg induced a significant increase in microvascular blood flow (*p = 0.04), while -125 mmHg did not significantly alter the microvascular blood flow., Conclusion: No hypoperfusion could be observed in the epicardium in neither normal nor ischemic myocardium during myocardial TNP.

Objective: : Hypothermia is known to limit the extent of myocardial infarction. The earlier hypothermia is applied to an ischemic myocardium, the more tissue can be salvaged. Topical negative pressure (TNP) is known to increase blood flow and stimulate angiogenesis in subcutaneous tissue and skeletal muscle. We have previously shown that a myocardial TNP of -50 mm Hg significantly increases microvascular blood flow in the underlying myocardium in ischemic and reperfused porcine myocardium. The present study was designed to elucidate the effect of different TNP levels on microvascular blood flow in reperfused myocardium during hypothermia., Methods: : Seven pigs underwent median sternotomy. The microvascular blood flow in the myocardium was recorded before and after the application of -50, -75, -100, -125, and -150 mm Hg using laser Doppler velocimetry. Analysis was performed in the epicardium and at a depth of 6 to 8 mm in the myocardium after 40 minutes of occlusion of the left anterior descending artery followed by cooling to 31°C, and reperfusion for another 20 minutes., Results: : A TNP of -50 mm Hg significantly increased blood the flow in the epicardium, from 116.7 ± 10.0 PU to 244.5 ± 52.6 PU (*P < 0.05) at 31°C. A TNP of -50 mm Hg significantly increased microvascular blood flow in the myocardium, from 155.0 ± 8.4 PU to 236.7 ± 61.5 PU (*P < 0.05)., Conclusions: : Only a TNP of -50 mm Hg, applied over the left anterior descending artery region in reperfused hypothermic myocardium significantly increased the microvascular blood flow in the epicardium and in the myocardium.

Background: Topical negative pressure (TNP) therapy has been adopted as a first-line treatment for wound healing. One of the mechanisms by which TNP improves healing is by stimulating blood flow to the wound edge. Among patients with ischemic heart disease, it is of great importance to improve the microvascular blood flow in the myocardium during episodes of ischemia to protect the myocardium from infarction. The present study was designed to elucidate the effect of TNP on microvascular blood flow in the myocardium., Methods: Six pigs underwent median sternotomy. The microvascular blood flow in the myocardium was recorded, before and after the application of TNP, by using laser Doppler velocimetry. Analyses were performed before left anterior descending artery (LAD) occlusion (normal myocardium), after 20 minutes of LAD occlusion (ischemic myocardium), and after 20 minutes of reperfusion (reperfused myocardium)., Results: TNP at -0 mm Hg increased microvascular blood flow in the normal myocardium from 14.7 +/- 3.9 perfusion units (PU) before to 25.8 +/- 6.1 PU after TNP application (p < 0.05), in the ischemic myocardium from 7.2 +/- 1.5 PU before to 13.8 +/- 2.6 PU after TNP application (p < 0.05), and in the reperfused myocardium from 10.8 +/- 2.0 PU before to 19.3 +/- 5.6 PU after TNP application (p < 0.05)., Conclusions: TNP increases the microvascular blood flow significantly in normal, ischemic, and reperfused myocardium and may provide a novel therapeutic tool in the treatment of ischemic myocardium.

Objectives: To assess the level of infectious complications and the impact of timing of a single, prophylactic, oral dose of ciprofloxacin 750 mg given either 2 hours before or in conjunction with ultrasound-guided core biopsy of the prostate in men without recognised risk factors and to analyse potential risk factors., Methods: All men undergoing prostate biopsy for elevated prostate specific antigen or clinical suspected prostate cancer were enrolled in an open, comparative prospective study. Excluded were men with recognised risk factors for infective complications. Two end points were chosen: febrile genitourinary infection and the results of postbiopsy urine culture., Results: A total of 1322 prostate biopsy occasions were made in 1157 men. Twelve (0.9%) cases of febrile genitourinary infections were recorded, two of which had proven sepsis. Administrating the drug 2 hours before or at the time of biopsy (p > 0.5) showed no statistical difference. Eight of 12 patients were shown to have prebiopsy undisclosed risk factors. Four percent developed postbiopsy, asymptomatic, significant bacteriuria. In addition, three (27%) men with prebiopsy unrecognised bacteriuria, who were accidentally enrolled, developed febrile genitourinary infection; one had proven sepsis., Conclusions: A single high-dose of oral ciprofloxacin 750 mg can be administered in direct conjunction with prostate biopsy to men without recognised risk factors, keeping the infection rate at approximately 1%. Bacteriuria before biopsy is a major risk factor for infective complications. Attention given to recognising individual risk factors would reduce the risk of infection further.

Monodelphis domestica (Marsupialia: Didelphidae) was used as a model animal to investigate and compare muscle adaptation to exercise training and cold exposure. The experimental treatment consisted of four groups of animals: either warm or cold acclimation temperature and with or without endurance exercise training. Maximal aerobic capacity during a running VO2max test in the warm-exercised or cold-exposed (with or without exercise) groups was about 130 mL O(2)/kg/min, significantly higher than the warm-acclimated controls at 113.5 mL O(2)/kg/min. Similarly, during an acute cold challenge (VO2summit), maximal aerobic capacity was higher in these three experimental groups at approximately 95 mL O(2)/kg/min compared with 80.4 mL O(2)/kg/min in warm-acclimated controls. Respiratory exchange ratio was significantly lower (0.89-0.68), whereas relative heart mass (0.52%-0.73%) and whole-body muscle mitochondrial volume density (2.59 to 3.04 cm(3)) were significantly higher following cold exposure. Chronic cold exposure was a stronger stimulus than endurance exercise training for tissue-specific adaptations. Although chronic cold exposure and endurance exercise are distinct challenges, physiological adaptations to each overlap such that the capacities for aerobic performance in response to both cold exposure and running are increased by either or both treatments.

Five cases of glycerol kinase deficiency are presented with clinical, biochemical, and genetic results. Two had the glycerol kinase deficiency as part of an Xp21 contiguous gene deletion syndrome-complex form-and three had an isolated form of the enzyme deficiency. In these we found two splice site mutations (IVS1+4A>G, IVS9-1G>T) and one insertion (1393&#95;1394insG). In patients with the complex form, a deletion of the DAX1, GK genes and the distal part of the DMD gene was found. A computerized study was performed to predict the effects of the splice site mutations. It showed that the IVS9-1G>T mutation substantially altered and removed the wild-type site and enhanced a cryptic site seven nucleotides downstream, and that the IVS1+4A>G diminished the strength of the wild-type donor site from strong to leaky. To verify these predictions, we developed an RT-PCR system with gene-specific primers that exclusively amplifies the Xp21 glycerol kinase gene transcript. Identification of individuals at risk is motivated by a need to avoid delay in a correct diagnosis. For reliable identification of heterozygotes for isolated glycerol kinase deficiency, knowledge of the specific mutation in the proband is required. This is easily obtained with the RT-PCR analyses developed in this study.

Background: The objective of this study was to determine if a chronic eccentric training intervention, i.e., negative work, could limit or even reverse sarcopenia and its related impairments and functional limitations. Is high-force eccentric training tolerable by elderly people and will it result in improved muscle size, strength, balance, and fall risk?, Methods: 21 frail elderly subjects (mean age, 80 years) experienced 11 weeks of lower extremity resistance training. The experimental eccentric (ECC) group (n=11) performed negative work while exercising on a high-force eccentric ergometer. The active "controls" performed traditional (TRAD) (n=10) lower extremity resistance exercises (weight training). Muscle fiber cross-sectional area and strength, balance, stair descending abilities, and fall risk were assessed prior to and following this intervention., Results: All ECC subjects who started the negative work intervention completed the study and reported the training to be relatively effortless; they experienced minimal and transient muscle soreness. Both groups experienced a significant increase in muscle fiber cross-sectional area (ECC=60%, TRAD=41%). Only the ECC group experienced significant improvements in strength (60%), balance (7%), and stair descent (21%) abilities. The timed up and go task improved in both groups, but only the ECC group went from a high to a low fall risk., Conclusions: These data demonstrate that lower extremity resistance exercise can improve muscle structure and function in those with limited exercise tolerance. The greater strength increase following negative work training resulted in improved balance, stair descent, and fall risk only in the ECC group. Because low energy cost is coupled to high force production with eccentric exercise, this intervention may be useful for a number of patients that are otherwise unable to achieve high muscle forces with traditional resistance exercise.

Recent recommendations in the National Cholesterol Education Program Expert Panel on Detection, Evaluation and Treatment of High Blood Cholesterol in Adults (ATPIII) are expected to increase the number of triglyceride (TG) determinations and consequently the risk of misinterpretation of "non-blanked" results with co-determination of free glycerol. Glycerol-kinase deficiency (GKD) is one cause of falsely elevated TG results. The natural history of isolated GKD with symptom-free cases and cases with e.g. severe episodes of hypoglycemia and/or ketoacidosis challenges the laboratories to identify cases of GKD and family members at risk. "Blanked" methods reporting both glycerol and TG concentration are therefore desirable. Molecular studies of the glycerol kinase (GK) and DAX1 genes were performed on four cases of "persistent hypertriglyceridemia" found in an Italian population and on two pediatric cases with high serum glycerol concentration. Two new missense mutations were found (C358Y, T961). Molecular modeling on GK from E. coli, indicate that these mutations are located in parts of the enzyme important for enzyme formation or activity. One splice-site mutation, (IVS9A-1G>A), was found in two brothers. Splice-junction analysis indicates that it destroys the splice site and results in a mixture of mRNA. Deletion of the GK and DAX1 genes was found in one child with symptoms of adrenal failure. A female with glycerolemia and glyceroluria had normal GK activity but possibly slightly decreased ability to oxidize glycerol.

One challenge for veterinarians, animal facilities and research scientists is the making of physiological estimates appropriate to a variety of species for which data are often either completely lacking or are incomplete. Our intent in compiling the data in this paper is to provide the best possible database of normal physiological and anatomical values primarily (though not exclusively) for four common mammalian model species: mouse, rat, dog and man. In order to make those data as accessible and applicable as possible, we have presented the results of this study in the form of body-size dependent allometric equations in which some variable (Y) is expressed as a dependent function of body mass (M) in the power-law equation, Y = aM(b). By compiling these data, it is apparent that the resultant equations are quantitatively grouped (with similar slope or 'b' values). These emergent patterns provide insights into body-size dependent 'principles of design' that seem to dictate several aspects of design and function across species among all mammals. In general, the weights of most individual organs scale as a constant fraction of body mass (i.e. the body mass exponent, b approximately equal to 1.0). Biological rates (e.g. heart rate, respiratory rate) scale as b approximately equal to -1/4. Finally, volume-rates (the product of volume and rate) such as cardiac output, ventilation and oxygen uptake vary as b approximately equal to 3/4.

Human endurance performance is often evaluated on the basis of the maximal rate of oxygen uptake during exercise (V(O(2)max)). Methods for overcoming limits to V(O(2)max) are touted as means for increasing athletic endurance performance. Here, we argue that the respiratory system is well designed for delivering O(2) to meet O(2) demands and that no single factor is rate-determining for O(2) uptake. We show that V(O(2)max) can vary 5000-fold among mammals, while any limitation to O(2) delivery by a single component of the respiratory system affects V(O(2)max) by 10% or less. Attempts to increase O(2) delivery by enhancing one step in the respiratory system are shown to have little effect. Blood doping, hyperoxia and O(2) supplementation of high-altitude natives all raise O(2) availability substantially to the working muscles, but these treatments increase V(O(2)max) only minimally. Finally, we argue that O(2) uptake is only one of a number of properties important to human aerobic performance.

Adrenocortical carcinoma (ACC) is a rare, aggressive tumor that is often detected in an advanced stage. Medical treatment with the adrenotoxic drug mitotane has been used for decades, but critical prospective trials on its role in residual disease or as an adjuvant agent after surgical resection are still lacking. The concept of a critical threshold plasma level of the drug must be confirmed in controlled studies. Because individual responsiveness cannot be predicted, the use mitotane is still advised for nonresectable disease. In case of cortisol or other steroid overproduction, several drugs (e.g., ketoconazole or aminoglutethimide) may be used. Chemotherapy with single agents (e.g., doxorubicin or cisplatin) have been disappointing, with low response rates (< 30%) and a short response duration. Part of this refractoriness may be explained by the fact that ACC tumors express the multidrug-resistance gene MDR-1. Chemotherapy with multiple agents has been tested in smaller series and has resulted in significant side effects. The best results were achieved by the combination of etoposide, doxorubicin, and cisplatin associated with mitotane, achieving a response rate of 54%, including individual complete responses. To be able to make progress in treating advanced ACC disease, adjuvant multicenter trials must be encouraged. When mitotane-based therapies are used, monitored drug levels are mandatory.

Substantial ATP supply by glycolysis is thought to reflect cellular anoxia in vertebrate muscle. An alternative hypothesis is that the lactate generated during contraction reflects sustained glycolytic ATP supply under well-oxygenated conditions. We distinguished these hypotheses by comparing intracellular glycolysis during anoxia to lactate efflux from muscle during sustained, aerobic contractions. We examined the tailshaker muscle of the rattlesnake because of its uniform cell properties, exclusive blood circulation, and ability to sustain rattling for prolonged periods. Here we show that glycolysis is independent of the O(2) level and supplies one-third of the high ATP demands of sustained tailshaking. Fatigue is avoided by rapid H(+) and lactate efflux resulting from blood flow rates that are among the highest reported for vertebrate muscle. These results reject the hypothesis that glycolysis necessarily reflects cellular anoxia. Instead, they demonstrate that glycolysis can provide a high and sustainable supply of ATP along with oxidative phosphorylation without muscle fatigue.

We inadvertently subjected a group of goats to 5 mo of cold exposure (mean minimum temperature less than -13 degrees C) during an experiment designed to examine the effects of training by daily running on one member of each sibling pair. During the three coldest months, the sedentary but cold-exposed goats experienced a 34% increase in maximal oxygen uptake (VO(2 max), P < 0.01) and a 29% increase in running speed at maximal (P < 0.05). When temperatures increased in the spring, both oxygen uptake and running speed decreased. We interpret these findings as evidence that cold is a sufficient stimulus to invoke the development of aerobic structures in muscle and that these structures subsequently can be utilized for the novel task of running. When the experiment was subsequently repeated without the cold exposure, running speed and VO(2 max) of trained animals increased less than in either group of cold-exposed animals. However, the cost of transport of these warm runners was lower than either group of cold-exposed animals (from 13-19%, P < 0. 0001). Thus, although aerobic capacity was increased with acclimation to severe winter weather, cold-acclimated goats operated with lower efficiency during locomotion.

NTBC treatment has greatly improved the survival of patients with acute tyrosinemia and has reduced the need for liver transplantation during early childhood. In patients in whom treatment with NTBC was started early in life, 2 cases (1%) of HCC have occurred during the first year of treatment, but no further cases have occurred among these patients, who have been followed for up to 9 years. In patients with late start of NTBC treatment, there is a considerable risk for liver malignancy. The risk for malignancy in this group of patients must be evaluated on an individual basis, taking into account the phenotype and clinical status of the patient. Porphyric crises are not seen in patients who comply with the medication regimen. NTBC is a well-tolerated drug with few adverse effects.

During locomotion, major muscle groups are often activated cyclically. This alternate stretch-shorten pattern of activity could enable muscle to function as a spring, storing and recovering elastic recoil potential energy. Because the ability to store and recover elastic recoil energy could profoundly affect the energetics of locomotion, one might expect this to be an adaptable feature of skeletal muscle. This study tests the hypothesis that chronic eccentric (Ecc) training results in a change in the spring properties of skeletal muscle. Nine female Sprague-Dawley rats underwent chronic Ecc training for 8 wk on a motorized treadmill. The spring properties of muscle were characterized by both active and passive lengthening force productions. A single "spring constant (Deltaforce/Deltalength) from the passive length-tension curves was calculated for each muscle. Results from measurements on long heads of triceps brachii muscle indicate that the trained group produced significantly more passive lengthening force (P = 0.0001) as well as more active lengthening force (P = 0.0001) at all lengths of muscle stretch. In addition, the spring constants were significantly different between the Ecc (1.71 N/mm) and the control (1.31 N/mm) groups. A stiffer spring is capable of storing more energy per unit length stretched, which is of functional importance during locomotion.

Lengthening (eccentric) muscle contractions are characterized by several unusual properties that may result in unique skeletal muscle adaptations. In particular, high forces are produced with very little energy demand. Eccentrically trained muscles gain strength, but the specific nature of fiber size and composition is poorly known. This study assesses the structural and functional changes that occur to normal locomotor muscle after chronic eccentric ergometry at training intensities, measured as oxygen uptake, that do not influence the muscle when exercised concentrically. Male subjects trained on either eccentric or concentric cycle ergometers for 8 wk at a training intensity starting at 54% and ending at 65% of their peak heart rates. The isometric leg strength increased significantly in the eccentrically trained group by 36%, as did the cross-sectional area of the muscle fiber by 52%, but the muscle ultrastructure remained unchanged. There were no changes in either fiber size, composition, or isometric strength in the concentrically trained group. The responses of muscle to eccentric training appear to be similar to resistance training.

Eccentric contractions, the lengthening of muscle while producing force, are a common part of our everyday movements. This study presents a challenge to the accepted notion that eccentric work causes obligatory muscle injury while demonstrating that an increase in muscle strength, via eccentric work, can occur with little demand for oxygen. Nine healthy subjects, ages 18-34, were randomly placed in either an eccentric or a concentric training group. Both groups trained for 6 wk while progressively increasing training frequency and duration. Significant gains in isometric leg strength were seen in the eccentrically trained subjects only. While training, the oxygen consumption required to do the eccentric work was equal to or less than that required to do the concentric work. The results demonstrate that by progressively increasing the eccentric work rate, significant isometric strength gains can be made without muscle injury and with minimal increase in metabolic demand for oxygen. The potential clinical implications of an eccentric training program that uncouples skeletal muscle strength improvements from the demand for oxygen are alluring.

In tyrosinaemia type I (McKusick 276700), fatal liver disease results either because of liver failure during infancy or early childhood or because of development of hepatocellular carcinoma during childhood or adolescence. This is caused by toxic metabolites which accumulate because of deficiency of fumarylacetoacetase, the last enzyme in the tyrosine catabolic pathway. NTBC is a potent inhibitor of 4-hydroxyphenylpyruvate dioxygenase and has been shown to efficiently prevent tyrosine degradation, and production of succinylacetone, in patients with tyrosinaemia. Since the first trial of NTBC treatment for tyrosinaemia type I in 1991, over 220 patients have been treated by the drug using a protocol which includes regular follow-up with reports of clinical and laboratory investigations to the study centre in Gothenburg, where additional analysis of critical variables is done on regularly collected samples. The course of the disease in patients with acute tyrosinaemia has changed dramatically. Only 10% of the patients have not clinically responded to NTBC treatment. In half of these patients, successful liver transplantation has been performed which has further reduced the mortality rate during infancy to 5%. The international NTBC study has now been going for 5 years and data have emerged that indicate a decreased risk for early development of hepatocellular carcinoma in patients who started treatment at an early age. There are now 101 patients aged 2-8 years who have started NTBC treatment before 2 years of age, and no cancer has developed after 2 years of age among these patients. However, there is no safe age with respect to occurrence of liver cancer, which has been recognized at diagnosis at 1 year of age in one patient and after a few months of treatment in an infant who was given NTBC at 5 months of age.

Adrenocortical carcinoma (ACC) is a rare disease with a poor prognosis. It has been difficult to establish a strict treatment program for ACC, and better treatment alternatives and diagnostic tools must be sought. Even though surgery is the treatment of choice, the role of surgery in advanced disease has been questioned. Eighteen consecutive patients were treated at our unit over a 22-year period (1975-1997). All patients underwent surgery and were followed by our protocol, which includes urinary steroid profiles, clinical examinations, analysis of steroid hormones, and radiologic investigations. Twelve patients received mitotane with drug concentration measurements to deliver an effective, nontoxic dose. The median duration of mitotane treatment was 12 months. Few side effects were observed. Four patients with low-stage tumors underwent second-look operations with no pathologic findings. Five patients were subjected to repeat operations, and the mean duration of the disease-free interval before repeat surgery for these patients was 59 months. There was a significant positive correlation between the disease-free interval and the observed survival after repeat surgery. Eleven patients with intentionally curative surgery had their urinary steroid profiles tested several times postoperatively. For five patients preoperative urine samples were also available. Steroid profiles indicated recurrent disease despite normal radiologic findings in two of these five patients. The follow-up ranged from 6 weeks to 24 years. The predicted 5-year survival was 58% according to the Kaplan-Meier method. We conclude that monitoring serum concentrations of mitotane makes long-term treatment possible with few side effects; steroid profile analysis can be used for early detection of tumor recurrence; and repeat surgery for recurrence is of value for patients with long disease-free intervals.

Skeletal muscle fibers are composed of three structural elements, each contributing a unique aspect of muscle function, yet each 'competing' in a sense for space inside the cell. The volume occupied by myofibrils determines the force of contraction, the volume of sarcoplasmic reticulum sets the rate of onset and relaxation of a fiber's contraction and hence contraction frequency, and the volume of mitochondria sets the level of sustained performance. The entirety of functional outcomes in muscle, from sustained isometric to high frequency contractions, and from high power output to high endurance, are all primarily attributable to shifts in the proportions (and relationships) of those three structures. This paper examines and reviews these components of muscle first to identify and summarize structure-function 'rules', and second to examine the balance between sometimes competing demands. In particular, we focus on those muscles in which power, endurance and frequency are all simultaneously high (flight muscles), and examine how muscle has 'solved' problems of space and energy demand. From these results and observations it would appear that for flight to have evolved in small animals, the double packing of inner mitochondrial membranes may be expected in animals under 50-80 g in mass, and asynchronous muscle is structurally essential for flight in small insects with wing beat frequencies above about 100 Hz.

Overlapping DNA fragments spanning approximately 21 kb of genomic DNA and encompassing the human 4-hydroxyphenylpyruvate dioxygenase gene (HPD) have been cloned by screening a human leukocyte genomic library and by PCR amplification of human fibroblastic DNA. A continuous gene sequence of 20,890 nucleotides was established, including 1957 bp of the 5'-flanking region. The 4-hydroxyphenylpyruvate dioxygenase gene is composed of 14 exons interrupted by 13 introns, all exhibiting conventional vertebrate splicing. Computer analysis of the DNA sequence revealed 12 complete repetitive Alu elements, 1 in the 5'-flanking region and 11 in the intervening segments of the gene. The transcriptional initiation site was mapped to a position 35 nt upstream of the translational start point. The computer analysis also identified several potential transcription regulatory elements, including one CRE site, two AP-2 sites, and two Sp1 sites, in the sequence upstream of the transcription initiation site. Functional analysis of promoter activity by transient transfection of chloramphenicolacetyl transferase reporter plasmids revealed a possible involvement of cyclic adenosine monophosphate in the regulation of transcription. The highest level of expression of 4-hydroxyphenylpyruvate dioxygenase was found in human liver tissue as demonstrated by Northern blot analysis.

To study the effect of carnitine depletion on physical working capacity, healthy subjects were administered pivaloyl-conjugated antibiotics for 54 days. The mean carnitine concentration in serum decreased from 35.0 to 3.5 mmicromol/L, and in muscle from 10 to 4.3 micromol/g noncollagen protein (NCP). Exercise tests were performed before and after 54 days' administration of the drug. At submaximal exercise, there was a slight increase in the concentration of 3-hydroxybutyrate in serum, presumably caused by decreased fatty acid oxidation in the liver. There was also a decreased consumption of muscle glycogen, indicating decreased glycolysis in the skeletal muscle. The muscle presumably had enough energy available, since there was no significant decrease in the concentration of adenosine triphosphate (ATP) and creatine phosphate during exercise. The work at maximal oxygen uptake (VO2max) and the maximal heart rate were reduced. Since VO2max is considered dependent on heart function, carnitine depletion seemed to affect cardiac function.

Sound production is one of the most energetically costly activities in animals. Minimizing contraction costs is one means of achieving the activation rates necessary for sound production (20-550 Hz) (refs 1-3) without exceeding energy supplies. Rattlesnakes produce a sustained, high-frequency warning sound by extremely rapid contraction of their tailshaker muscles (20-90 Hz) (refs 4,5). The ATP cost per twitch is only 0.015 micromol ATP per g muscle per twitch during rattling, as measured by in vivo magnetic resonance. The reduced volume density of myofibre (32%) in tailshaker muscle is consistent with contraction cost being minimized (crossbridge cycling), in contrast to the contractile costs of vertebrate locomotory and asynchronous insect flight muscle. Thus tailshaker muscle is an example of sound-producing muscle designed for 'high frequency, minimal cost'. The high rates of rattling are achieved by minimizing contractile use of ATP, which reduces the cost per twitch to among the lowest found for striated muscle.

Vertebrate sound producing muscles often operate at frequencies exceeding 100 Hz, making them the fastest vertebrate muscles. Like other vertebrate muscle, these sonic muscles are "synchronous," necessitating that calcium be released and resequestered by the sarcoplasmic reticulum during each contraction cycle. Thus to operate at such high frequencies, vertebrate sonic muscles require extreme adaptations. We have found that to generate the "boatwhistle" mating call (approximately 200 Hz), the swimbladder muscle fibers of toadfish have evolved (i) a large and very fast calcium transient, (ii) a fast crossbridge detachment rate, and (iii) probably a fast kinetic off-rate of Ca2+ from troponin. The fibers of the shaker muscle of rattlesnakes have independently evolved similar traits, permitting tail rattling at approximately 90 Hz.

The western diamondback rattlesnake Crotalus atrox can rattle its tail continuously for hours at frequencies approaching 90 Hz. We examined the basis of these fast sustainable contractions using electromyography, data on oxygen uptake and the quantitative ultrastructure of the tailshaker muscle complex. The tailshaker muscle has no apparent unique structures; rather, the relative proportions of the structures common to all skeletal muscles appear to be present (1) to minimize activation, contraction and relaxation times via an extremely high volume density of sarcoplasmic reticulum (26 %) as well as, (2) to maximize ATP resysnthesis via a high volume density of mitochondria (26 %). The high rate of ATP supply is reflected in the in vivo muscle mass-specific oxygen uptake of this group of muscles which, at 585 ml O2 kg-1 min-1 during rattling at 30 &deg;C body temperature, exceeds that reported for other ectotherm and many endotherm muscles. Since the change in oxygen uptake paralleled that of the rattling frequency over the range of measured body temperatures, there was a nearly constant O2 cost per muscle contraction (0.139&plusmn;0.016 &micro;l O2 g-1). Electromyo-graphic analysis suggests that each of the six muscles that make up the shaker complex may be a single motor unit. Finally, the maximum rate of mitochondrial oxygen uptake is similar to that of various mammals, a hummingbird, a lizard, an anuran amphibian and of isolated mitochondria (at 10 000-40 000 molecules O2 s-1 &micro;m2 of cristae surface area, when normalized to 30 &deg;C), suggesting a shared principle of design of the inner mitochondrial membrane among the vertebrates.

Objective: To study the effect of induced carnitine depletion on myocardial structure and function., Subjects and Design: 7 healthy adult volunteers given 1200 mg pivmecillinam per day for 7-8 weeks were studied by echocardiography before and after 7-8 weeks of treatment and a 15 months follow up after the treatment period., Setting: Teaching hospital., Main Outcome Measures: Carnitine concentration in serum, urine, and muscle and echocardiographic measurements., Results: After 7-8 weeks of treatment the median free serum carnitine concentration was reduced to 7% and the median total muscle carnitine concentration to 46% of the pretreatment levels. The median diastolic interventricular septum thickness decreased by 14% (mean 26%, P = 0.028) and the median left ventricular mass by 10% (mean 20%, P = 0.018). Fifteen months later these dimensions had increased but not completely returned to pretreatment values., Conclusions: Extended treatment with pivalic acid containing antibiotics causes carnitine depletion which may lead to changes in cardiac structure.

Hereditary tyrosinaemia type I, a severe autosomal recessive metabolic disease, affects the liver and kidneys and is caused by deficiency of fumarylacetoacetate hydrolase (FAH). Mice homozygous for a FAH gene disruption have a neonatal lethal phenotype caused by liver dysfunction and do not represent an adequate model of the human disease. Here we demonstrate that treatment of affected animals with 2-(2-nitro-4-trifluoro-methylbenzyol)-1,3-cyclohexanedione abolished neonatal lethality, corrected liver function and partially normalized the altered expression pattern of hepatic mRNAs. The prolonged lifespan of affected animals resulted in a phenotype analogous to human tyrosinaemia type I including hepatocellular carcinoma. The adult FAH-/- mouse will serve as useful model for studies of the pathophysiology and treatment of hereditary tyrosinaemia type I as well as hepatic cancer.

Treatment with pivalic acid containing prodrugs has been shown to cause carnitine depletion by loss of pivaloyl carnitine in urine. A 7-day standard pivmecillinam treatment of adults lead to a marked decrease of the free serum carnitine concentration (44.6 to 12.9 mumol/liter), whereas no change was seen in those given norfloxacine (40.0 to 40.5 mumol/liter). In some patients irrespective of age the free serum carnitine concentration was decreased to levels (around 10 mumol/liter) at which an impaired ketone-body production may occur. Therefore, there is reason for cautious use of this type of drug irrespective of the age of the patients.

Some 20 compartments of the body may be measured by CT and organ areas determined in 28 CT scans. Advantages of CT are described. While there have been extensive studies of hormones in pre- and postnatal growth, apart from evidence from disease, the role of hormones in adults has been less known. Data on growth hormone and sex hormones, from organ-oriented body composition studies, are summarized, together with implications for the relation between body composition and cardiovascular risk. Sex-specific anthropometric equations allow estimation of LBM, visceral and sc AT with <20% error. In the obese such estimates show visceral AT to be a stronger risk predictor than other compartments or W/HR.

In a double-blind, placebo-controlled trial, the effects of recombinant human growth hormone were studied on cerebrospinal fluid concentrations of growth hormone, insulin-like growth factor 1 (IGF-1), insulin-like growth factor binding protein-3 (IGFBP-3), monoamine metabolites, neuropeptides and endogenous opioid peptides. Twenty patients, 10 patients in each of 2 groups, with adult-onset, growth hormone deficiency were treated for 1 month with recombinant human growth hormone (0.25 U/kg/week) or placebo. All the patients received the appropriate thyroid, adrenal and gonadal hormone replacement. In cerebrospinal fluid, the mean concentration of growth hormone increased from 13.3 +/- 4.4 to 149.3 +/- 22.2 muU/l (p = 0.002), during recombinant human growth hormone treatment. The cerebrospinal fluid IGF-I concentration increased from 0.67 +/- 0.04 to 0.99 +/- 0.10 micrograms/l (p = 0.005) and the IGFBP-3 concentration rose from 13.4 +/- 1.25 to 17.5 +/- 1.83 micrograms/l (p = 0.002). The dopamine metabolite homovanillic acid decreased from 282.1 +/- 36.0 to 234.3 +/- 26.5 nmol/l (p = 0.02) and the vasoactive intestinal peptide decreased from 4.1 +/- 0.6 to 3.7 +/- 0.4 pmol/l (p = 0.03). Cerebrospinal fluid immunoreactive beta-endorphin increased from 24.4 +/- 1.8 to 29.9 +/- 2.1 pmol/l (p = 0.002). There were no significant changes compared to baseline in the cerebrospinal fluid concentrations of enkephalins, dynorphin A, the norepinephrine metabolite 3-methoxy-4-hydroxyphenyl-ethyleneglycol, the serotonin metabolite 5-hydroxyindoleacetic acid, gamma-aminobutyric acid, somatostatin or corticotropin-releasing factor.(ABSTRACT TRUNCATED AT 250 WORDS)

One source of muscle fatigue may be the failure to provide the required oxygen by any step in the oxygen transport cascade or a lack of the necessary machinery to utilize that oxygen. We favor abandoning the concept of a single rate-limiting step for the concept of tuned resistors, each contributing to the overall resistance to oxygen flow. However, because some of these steps have considerably less phenotypic plasticity than others, these are the component parts of the respiratory system that must be built with adequate "reserve" to accommodate adaptive increases in the other steps (Lindstedt et al., 1988; Weibel et al., 1992; Lindstedt et al., 1994). These structures will usually appear to be over built except in those rare individual animals at the species-specific limit of VO2 in which these less malleable structures may be limiting.

Using a panel of human-rodent somatic cell hybrids, we have previously mapped the gene (HPD, previously called PPD) encoding 4-hydroxyphenylpyruvate dioxygenase to the distal half of the long arm of human chromosome 12, region q14-->qter. To obtain a genomic probe useful for fluorescence in situ hybridization (FISH) analysis we screened a human leukocyte genomic library and isolated a 13.4-kb phage clone, which by restriction fragment and sequence analyses was shown to contain exons 1-10 of HPD and approximately 2-kb upstream sequences. We now report the subregional localization of HPD to 12q24-->qter based on two color FISH analysis employing this clone.

The mean minimal transit time for blood in muscle capillaries (tc) was estimated in six species, spanning two orders of magnitude in body mass and aerobic capacity: horse, steer, dog, goat, fox and agouti. Arterial (CaO2) and mixed venous (CvO2) blood O2 concentrations, blood hemoglobin concentrations ([Hb]) and oxygen uptake rates were measured while the animals ran on a treadmill at a speed that elicited the maximal oxygen consumption rate (VO2max) from each animal. Blood flow to the muscles (Qm) was assumed to be 85% of cardiac output, which was calculated using the Fick relationship. Total muscle capillary blood volume (Vc) and total muscle mitochondrial volume were estimated by morphometry, using a whole-body muscle sampling scheme. The tc was computed as Vc/Qm. The tc was 0.3-0.5 s in the 4 kg foxes and agoutis, 0.7-0.8 s in the 25 kg dogs and goats, and 0.8-1.0 s in the 400 kg horses and steers. The tc was positively correlated with body mass and negatively correlated with transcapillary O2 release rate per unit capillary length. Mitochondrial content was positively correlated with VO2max and with the product of Qm and [Hb]. These data suggested that Qm, Vc, maximal hemoglobin flux, and consequently tc, are co-adjusted to result in muscle O2 supply conditions that are matched to the O2 demands of the muscles at VO2max.

Long-term treatment with pivalic acid prodrug results in impaired ketone-body production. Therefore, it was of interest to investigate whether short-term treatment had any influence on the fatty acid oxidation. In this study six healthy males were given 1200 mg per day of pivmecillinam for 12 days to induce carnitine deficiency. The concentration of free carnitine in serum was reduced from a mean of 42.8 mumol/liter (range, 31-48) to 11.6 mumol/liter (range, 7.0-24), but the muscle carnitine concentration was not reduced. A 36-h fasting test was performed before and after drug administration to study the effect on ketone-body production. After treatment, the two subjects with the lowest level of serum free carnitine at the end of the fasting period had impaired ketogenesis. This indicates a carnitine deficiency in the liver which was reflected in the free carnitine concentration for by mobilization of muscle carnitine. We conclude that there is a substantial risk to develop carnitine deficiency and impaired fatty acid oxidation in the liver during short-term treatment with drugs conjugated with pivalic acid.

We have examined aerobic exercise performance among the mammals with particular attention to the constraints that body size places on all aspects of muscle biomechanics, aerobic energetics, tissue oxygen diffusion, cardiovascular oxygen delivery, and pulmonary oxygen uptake. Several body-size-dependent patterns emerge that seemingly govern aerobic performance in mammals, with the caveat that at any given body size there is a range of aerobic capacities, the result of natural selection operating on the size-dependent "default values" of structure and function. Among these default values, the following apparent functional clusters surface: 1. In general, concentrations and pressures (e.g., of proteins and gases) are roughly independent of body size. Inspiratory and expiratory ventilation pressures, blood pressure and the partial pressures of O2 and CO2 in lungs, blood, and tissues do not vary with body size. Likewise, concentrations of hemoglobin, myoglobin, and hematocrit are independent of body size. 2. Most volumes and capacities scale linearly with body size (i.e., as a constant function of body mass). In addition to heart, lung, and total blood volumes, important examples relevant to exercise performance are the diffusing capacities for oxygen in the lung and, apparently, in the tissues. 3. Finally, most time-dependent variables related to oxygen delivery scale allometrically with body mass; they are of shorter duration in small animals than in large ones. Biological rates, for example, Vmax of working muscle, heart and respiratory rates, and transit times of blood through the muscles and lungs, all vary roughly as the -1/5 to -1/4 power of body mass.

A 21 month old girl presented with a short history of frequent falls and a right sided foot drop. She went on to suffer recurrent episodes of distal weakness in her arms and legs with hyporeflexia. Electrophysiological studies were consistent with inflammatory demyelinating polyradiculoneuropathy (IDP) and treatment with corticosteroids appeared to lead to an improvement. However, the development of hypertension, evidence of tubulopathy, and hepatomegaly led to re-evaluation. A diagnosis of type I tyrosinaemia was made, based on increased urinary excretion of succinylacetone and decreased activity of fumarylacetoacetase in her cultured skin fibroblasts. A low tyrosine diet did not prevent life-threatening exacerbations of neuropathy but intravenous haemarginate appeared to aid her recovery from one exacerbation. An immediate improvement in strength was seen after starting treatment with 2-(2-nitro-4-trifluoro-methyl-benzoyl)-1,3-cyclohexanedione (NTBC), an inhibitor of 4-hydroxy-phenylpyruvate dioxygenase. A liver transplant was performed but the patient died of immediate postoperative complications. Tyrosinaemia needs to be considered in a child with recurrent peripheral neuropathy because (i) the signs of liver disease and renal tubular dysfunction may be subtle; (ii) acute exacerbations may be life threatening; (iii) specific forms of treatment are available.

This part of an on-going intervention trial analyses impacts of obesity on psychosocial factors and health. The study sample comprised 800 obese men (BMI > or = 34 kg/m2) and 943 women (BMI > or = 38 kg/m2) ranging in age from 37 to 57 years. All participants completed standardized health-related quality of life measures, a validated obesity-specific eating inventory and study-specific questionnaires on current and past health status, use of medical care and medications, socioeconomic status, dietary habits, physical activity habits, weight history and familial history of obesity. Chronic patients and population samples were used as reference. The obese reported distinctly poorer current health and less positive mood states than the reference subjects, women being worse than men. Anxiety and/or depression on a level indicating psychiatric morbidity were more often seen in the obese and again women reported more affliction than men. Furthermore, the average poor mental well-being was worst than in chronically ill or injured patients, such as rheumatoid, cancer survivors and spinal cord injured persons. Predictors of perceived health and psychosocial functioning could be discerned using a comprehensive system of statistical analyses (16-28% explained variance). A background of both somatic and psychiatric morbidity was decisive for the health and psychosocial functioning in the obese; joint symptoms and angina pectoris dominated among somatic variables. Physical inactivity was the most prominent of traditional risk factors. The number of dieting attempts and body image were important weight correlates. Our results provide further evidence to the effect that severe obesity is a crippling condition.

We report the primary structure of 4-hydroxyphenylpyruvate dioxygenase [4-hydroxyphenyl-pyruvate:oxygen oxidoreductase (hydroxylating, decarboxylating)]. The work is based on the isolation of cDNA clones from human liver lambda gt11 libraries. Several overlapping clones covering the coding sequence were characterized. In parallel, peptides from four different digests of the purified protein were analysed for their amino-acid sequence. These peptide sequences covered 86% of the cDNA-derived amino-acid sequence. This gives the sequence for a polypeptide of 392 amino acids with a calculated molecular mass of 44.8 kDa. There is more than 80% identity between the human and the pig enzymes and also between these enzymes and the F antigen from rat and the two allelic forms of this antigen from mouse. The enzyme has 53% conserved amino acids and 27% identical amino acids in common with 4-hydroxyphenylpyruvate dioxygenase from Pseudomonas sp. P.J. 874 and 52% conserved and 28% identical residues, with a protein from Shewanella colwelliana. At the C-terminus there is 61% identity between the seven proteins. These results indicate that these proteins are all 4-hydroxyphenylpyruvate dioxygenases. The identity of the C-terminus makes this part of the molecule a candidate for a functional role in the catalytic process. At conserved positions in all seven enzymes, there are two tyrosine residues and three histidine residues, i.e. amino acids which have been implicated as ligands for iron in 2-oxoacid-dependent dioxygenases. The gene encoding the enzyme was localized to chromosome 12q14-->qter by Southern-blot analysis of human-rodent somatic-cell hybrids.

gamma-Butyrobetaine hydroxylase is a 2-oxoglutarate-dependent dioxygenase that catalyzes the hydroxylation of gamma-butyrobetaine to carnitine, the last step in the biosynthesis of carnitine from lysine. The primary structure of the enzyme from Pseudomonas sp. AK1 has been determined. Sequence analysis of the intact protein and of peptides from essentially three different digests established the presence of a peptide chain containing 383 residues, and an N-terminal truncated form of 382 residues. The two chains have molecular masses of 43,321 Da and 43,207 Da, respectively, and are identical except for the presence or absence of an N-terminal asparagine residue; the shorter form starts with an alanine residue. In preparations of the dimeric protein, the two chains occur in an approximate ratio of 1:1. There are nine cysteine residues and 13 histidine residues, i.e. amino acids which have been postulated as ligands for iron binding. In spite of functional similarities, there appears to be no clear sequence similarities with any of the other mammalian 2-oxoglutarate-dependent dioxygenases so far characterized.

Objective: To evaluate the results of treatment of a consecutive series of patients with adrenocortical carcinoma who presented during the six year period 1985 to 1991., Design: Open study., Setting: Departments of Surgery, Pathology, Endocrinology, and Clinical Chemistry, Sahlgren Hospital, Göteborg, Sweden., Subjects: 10 consecutive patients, two with recurrent and eight with primary adrenocortical carcinoma., Interventions: All patients were treated surgically. Two required preoperative embolisation of the tumour vessels to facilitate excision of particularly large tumours, and eight were given adjuvant treatment with mitotane (o,p'-DDD)., Results: At a median follow up of 1.5 years (range 3 months, to 21 years) 6 patients were alive with no radiological or biochemical signs of disease; 2 were alive, but with signs of recurrence (at 3 months and 6 years, respectively); and two had died of their disease (at 4 and 8 months, respectively). For the past two years all patients have had their urinary steroid profiles monitored by gas chromatography and mass spectrometry to detect recurrence of the tumour at the earliest possible stage., Conclusion: Operation is the treatment of choice for patients with adrenocortical carcinoma, particularly stages I-III. The role of mitotane as adjuvant treatment can be evaluated only in multicentre studies.

Body size fundamentally affects maximal locomotor performance in mammals. Comparisons of performances of different-sized animals yield different results if made using relative, rather than absolute scales. Absolute speed may be a reasonable way to evaluate the locomotor performance of an animal that must escape predators in real time. However, comparisons of metabolic power in animals of different size can only be made meaningfully on a mass-specific basis. Numerous factors associated with the mechanics, energetics, and storage of elastic energy during locomotion change with body size, which results in allometric relationships that make the energetic cost of locomotion (alpha Mb-0.3) more expensive for small mammals than for large mammals. Small mammals have lower enzymatic capacities for anaerobic glycolysis (alpha Mb0.15) and higher specific aerobic capacities (alpha Mb-0.13) than large mammals. However, the energetic cost of transport increases more than aerobic power as mammals get smaller. The higher ratio of cost to available power in small mammals may explain why they run more slowly than large mammals, as a rule. Maximum aerobic capacity is allometrically related to body size. Limits to VO2max can be imposed by mitochondrial oxidative capacity, as in goats, or by the O2 transport system, as in humans and horses. No single step in the O2 transport system can limit the flux of O2 by itself; however, in an average non-athletic species of mammal, any of the steps in the system might appear to be the weakest link. In highly aerobic athletic species, and possibly elite athletic individuals of other species (e.g. humans), the malleable elements of the O2 transport system may develop to the point that their O2 transport capacities approach that of the least malleable element in the system, the lung. VO2max is very high in such individuals, and appears to be limited by simultaneous failure of all components of the O2 transport system.

We report the finding of mitochondrial ATP-synthase deficiency in a child with persistent 3-methylglutaconic aciduria. The child presented in the neonatal period with severe lactic acidosis, which was controlled by Na-HCO3 and glucose infusions. During the 1st y of life, there were several episodes of lactic acidosis precipitated by infections or prolonged intervals between meals. The excretion of lactate in urine was variable, but there was a persistent high excretion of 3-methylglutaconic acid. The activity of 3-methylglutaconyl-CoA hydratase in fibroblasts was normal. The child had a hypertrophic cardiomyopathy and magnetic resonance images revealed hypoplasia of corpus callosum. The gross motor and mental development was retarded, but there were no other neurologic signs. Investigation of muscle mitochondrial function at 1 y of age revealed a severe mitochondrial ATP-synthase deficiency (oligomycin-sensitive, dinitrophenol-stimulated Mg2+ ATPase activity: 27 nmol x min-1 x (mg protein)-1, control range 223-673 nmol x min-1 x (mg protein)-1. The mitochondrial respiratory rate was low and tightly coupled. The respiratory rate was normalized by the addition of an uncoupler. Low Mg2+ ATPase activity was also demonstrated by histochemical methods. Morphologic examination revealed ultrastructural abnormalities of mitochondria. There was no deletion of mitochondrial DNA. The sequences of the ATP synthase subunit genes of mitochondrial DNA were in accordance with published normal sequences.