Time has come to include Human Papillomavirus (HPV) testing in sperm donor banks.

HPV is well known as a potential cause of cervical cancer. Less well known is its link to temporal subfertility that is caused by binding of infectious virions to the spermatozoa's head which induces sperm-DNA damage and causes a reduction in clinical pregnancy rates in women receiving HPV positive semen. This impact on the global fertility burden remains greatly underestimated and underexplored. This risk of reduced fertility due to infectious HPV in sperm is especially important when donor sperm insemination is considered, since testing for the presence of HPV virions before use seems warranted. We tested 514 donor sperm samples from 3 different sperm banks for 18 different HPV types. Overall 3.9% (20/514) of tested donor sperm was positive for HPV, with different prevalence among the 3 different sperm banks (3.6% bank A, 3.1% bank B and 16.7% bank C). Also the HPV virion per spermatozoon ratio in donor samples was similar across the different sperm banks (95% CI 0,01 to 1,07 HPV virions/spermatozoon). When HPV positive donor sperm was used, no clinical pregnancies resulted, whereas when HPV negative donor sperm was used the clinical pregnancy rate was 14.6%. From both a cost/benefit and a safety point of view we recommend that donor sperm should always be tested for HPV before using it for insemination.

Magnetic resonance imaging of the proximal upper extremity musculature in boys with Duchenne muscular dystrophy.

There is a pressing need for biomarkers and outcomes that can be used across disease stages in Duchenne muscular dystrophy (DMD), to facilitate the inclusion of a wider range of participants in clinical trials and to improve our understanding of the natural history of DMD. Quantitative magnetic resonance imaging (qMRI) and spectroscopy (MRS) biomarkers show considerable promise in both the legs and forearms of individuals with DMD, but have not yet been examined in functionally important proximal upper extremity muscles such as the biceps brachii and deltoid. The primary objective of this study was to examine the feasibility of implementing qMRI and MRS biomarkers in the proximal upper extremity musculature, and the secondary objective was to examine the relationship between MR measures of arm muscle pathology and upper extremity functional endpoints. Biomarkers included MRS and MRI measures of fat fraction and transverse relaxation time (T 2). The MR exam was well tolerated in both ambulatory and non-ambulatory boys. qMR biomarkers differentiated affected and unaffected participants and correlated strongly with upper extremity function (r = 0.91 for biceps brachii T 2 versus performance of upper limb score). These qMR outcome measures could be highly beneficial to the neuromuscular disease community, allowing measurement of the quality of functionally important muscles across disease stages to understand the natural history of DMD and particularly to broaden the opportunity for clinical trial participation.

Bone loss in a new rodent model combining spinal cord injury and cast immobilization.

OBJECTIVES: Characterize bone loss in our newly developed severe contusion spinal cord injury (SCI) plus hindlimb immobilization (IMM) model and determine the influence of muscle contractility on skeletal integrity after SCI. METHODS: Female Sprague-Dawley rats were randomized to: (a) intact controls, (b) severe contusion SCI euthanized at Day 7 (SCI-7) or (c) Day 21 (SCI-21), (d) 14 days IMM-alone, (e) SCI+IMM, or (f) SCI+IMM plus 14 days body weight supported treadmill exercise (SCI+IMM+TM). RESULTS: SCI-7 and SCI-21 exhibited a >20% reduction in cancellous volumetric bone mineral density (vBMD) in the hindlimbs (p⋜0.01), characterized by reductions in cancellous bone volume (cBV/TV%), trabecular number (Tb.N), and trabecular thickness. IMM-alone induced no observable bone loss. SCI+IMM exacerbated cancellous vBMD deficits with values being >45% below Controls (p⋜0.01) resulting from reduced cBV/TV% and Tb.N. SCI+IMM also produced the greatest cortical bone loss with distal femoral cortical area and cortical thickness being 14-28% below Controls (p⋜0.01) and bone strength being 37% below Controls (p⋜0.01). SCI+IMM+TM partially alleviated bone deficits, but values remained below Controls. CONCLUSIONS: Residual and/or facilitated muscle contractility ameliorate bone decrements after severe SCI. Our novel SCI+IMM model represents a clinically-relevant means of assessing strategies to prevent SCI-induced skeletal deficits.

Gene transfer of arginine kinase to skeletal muscle using adeno-associated virus.

In this study, we tested the feasibility of non-invasively measuring phosphoarginine (PArg) after gene delivery of arginine kinase (AK) using an adeno-associated virus (AAV) to murine hindlimbs. This was achieved by evaluating the time course, regional distribution and metabolic flux of PArg using (31)phosphorus magnetic resonance spectroscopy ((31)P-MRS). AK gene was injected into the gastrocnemius of the left hindlimb of C57Bl10 mice (age 5 weeks, male) using self-complementary AAV, type 2/8 with desmin promoter. Non-localized (31)P-MRS data were acquired over 9 months after injection using 11.1-T and 17.6-T Bruker Avance spectrometers. In addition, (31)P two-dimensional chemical shift imaging and saturation transfer experiments were performed to examine the spatial distribution and metabolic flux of PArg, respectively. PArg was evident in each injected mouse hindlimb after gene delivery, increased until 28 weeks, and remained elevated for at least 9 months (P<0.05). Furthermore, PArg was primarily localized to the injected posterior hindimb region and the metabolite was in exchange with ATP. Overall, the results show the viability of AAV gene transfer of AK gene to skeletal muscle, and provide support of PArg as a reporter that can be used to non-invasively monitor the transduction of genes for therapeutic interventions.

Longitudinal measurements of MRI-T2 in boys with Duchenne muscular dystrophy: effects of age and disease progression.

Duchenne muscular dystrophy (DMD) is characterized by an increased muscle damage and progressive replacement of muscle by noncontractile tissue. Both of these pathological changes can lengthen the MRI transverse proton relaxation time (T2). The current study measured longitudinal changes in T2 and its distribution in the lower leg of 16 boys with DMD (5-13years, 15 ambulatory) and 15 healthy controls (5-13years). These muscles were chosen to allow extended longitudinal monitoring, due to their slow progression compared with proximal muscles in DMD. In the soleus muscle of boys with DMD, T2 and the percentage of pixels with an elevated T2 (⩾2SD above control mean T2) increased significantly over 1year and 2years, while the width of the T2 histogram increased over 2years. Changes in soleus T2 variables were significantly greater in 9-13years old compared with 5-8years old boys with DMD. Significant correlations between the change in all soleus T2 variables over 2years and the change in functional measures over 2years were found. MRI measurement of muscle T2 in boys with DMD is sensitive to disease progression and shows promise as a clinical outcome measure.

Non-invasive assessment of lower extremity muscle composition after incomplete spinal cord injury.

STUDY DESIGN: Cross-sectional study. OBJECTIVE: (1) To quantify intramyocellular lipid (IMCL) content of the soleus muscle. (2) To assess the T(2) relaxation rates in the lower extremity skeletal muscles in persons with incomplete spinal cord injury (SCI). SETTING: Academic Institution, Florida. METHODS: Eight subjects (42+/-10 years old; 70+/-12 kg; 176+/-10 cm) with chronic (17+/-9 months post injury) motor SCI (C4-T12; ASIA C or D) and eight matched healthy controls were tested. Localized unsuppressed proton spectroscopy (H-MRS) was performed to estimate total lipid content and individual lipid components; IMCL and extramyocellular lipid (EMCL) from the soleus muscle. T(2)-weighted imaging of lower extremity muscles yielded muscle T(2) rates. RESULTS: The IMCL content of the soleus muscle was 3.3 times higher in the patient group as compared to controls (P=0.002; 0.0401 (0.0234-0.0849) versus 0.0123 (0.0090-0.0175)). Similarly, EMCL measures were 4.5 times higher as compared to the controls (P=0.002). Significant differences were observed in the T(2) relaxation times of the soleus and gastrocnemius muscles (P<0.05). CONCLUSION: The increased levels of IMCL might interfere with the glucose uptake in skeletal muscle; potentially predisposing persons with incomplete SCI to the development of peripheral insulin resistance. Marked elevations in the T(2) relaxation times of the locomotor muscles are reflective of an altered muscle composition.

A longitudinal study of skeletal muscle following spinal cord injury and locomotor training.

STUDY DESIGN: Experimental rat model of spinal cord contusion injury (contusion SCI). OBJECTIVE: The objectives of this study were (1) to characterize the longitudinal changes in rat lower hindlimb muscle morphology following contusion SCI by using magnetic resonance imaging and (2) to determine the therapeutic potential of two types of locomotor training, treadmill and cycling. SETTING: University research setting. METHODS: After moderate midthoracic contusion SCI, Sprague-Dawley rats were assigned to either treadmill training, cycle training or an untrained group. Lower hindlimb muscle size was examined prior to SCI and at 1-, 2-, 4-, 8-, and 12-week post injury. RESULTS: Following contusion SCI, we observed significant atrophy in all rat hindlimb muscles with the posterior muscles (triceps surae and flexor digitorum) showing greater atrophy than the anterior muscles (tibialis anterior and extensor digitorum). The greatest amount of atrophy was measured at 2-week post injury (range from 11 to 26%), and spontaneous recovery in muscle size was observed by 4 weeks post-SCI. Both cycling and treadmill training halted the atrophic process and accelerated the rate of recovery. The therapeutic influence of both training interventions was observed within 1 week of training and no significant difference was noted between the two interventions, except in the tibialis anterior muscle. Finally, a positive correlation was found between locomotor functional scores and hindlimb muscle size following SCI. CONCLUSIONS: Both treadmill and cycle training diminish the extent of atrophy and facilitate muscle plasticity after contusion SCI.

Resistance training and locomotor recovery after incomplete spinal cord injury: a case series.

STUDY DESIGN: Longitudinal intervention case series. OBJECTIVE: To determine if a 12-week resistance and plyometric training program results in improved muscle function and locomotor speed after incomplete spinal cord injury (SCI). SETTING: University research setting. METHODS: Three ambulatory individuals with chronic (18.7+/-2.2 months post injury) motor incomplete SCI completed 12 weeks of lower extremity resistance training combined with plyometric training (RPT). Muscle maximum cross-sectional area (max-CSA) of the knee extensor (KE) and plantar flexor (PF) muscle groups was determined using magnetic resonance imaging (MRI). In addition, peak isometric torque, time to peak torque (T (20-80)), torque developed within the initial 220 ms of contraction (torque(220)) and average rate of torque development (ARTD) were calculated as indices of muscle function. Maximal as well as self-selected gait speeds were determined pre- and post-RPT during which the spatio-temporal characteristics, kinematics and kinetics of gait were measured. RESULTS: RPT resulted in improved peak torque production in the KE (28.9+/-4.4%) and PF (35.0+/-9.1%) muscle groups, as well as a decrease in T(20-80), an increased torque(220) and an increase ARTD in both muscle groups. In addition, an increase in self-selected (pre-RPT=0.77 m/s; post-RPT=1.03 m/s) and maximum (pre-RPT=1.08 m/s; post-RPT=1.47 m/s) gait speed was realized. Increased gait speeds were accompanied by bilateral increases in propulsion and hip excursion as well as increased lower extremity joint powers. CONCLUSIONS: The combination of lower extremity RPT can attenuate existing neuromuscular impairments and improve gait speed in persons after incomplete SCI.

A quantitative study of bioenergetics in skeletal muscle lacking carbonic anhydrase III using 31P magnetic resonance spectroscopy.

Oxidative slow skeletal muscle contains carbonic anhydrase III in high concentration, but its primary function remains unknown. To determine whether its lack handicaps energy metabolism and/or acid elimination, we measured the intracellular pH and energy phosphates by (31)P magnetic resonance spectroscopy in hind limb muscles of wild-type and CA III knockout mice during and after ischemia and intense exercise (electrical stimulation). Thirty minutes of ischemia caused phosphocreatine (PCr) to fall and P(i) to rise while pH and ATP remained constant in both strains of mice. PCr and P(i) kinetics during ischemia and recovery were not significantly different between the two genotypes. From this we conclude that under neutral pH conditions resting muscle anaerobic metabolism, the rate of the creatine kinase reaction, intracellular buffering of protons, and phosphorylation of creatine by mitochondrial oxygen metabolism are not influenced by the lack of CA III. Two minutes of intense stimulation of the mouse gastrocnemius caused PCr, ATP, and pH to fall and ADP and P(i) to rise, and these changes, with the exception of ATP, were all significantly larger in the CA III knockouts. The rate of return of pH and ADP to control values was the same in wild-type and mutant mice, but in the mutants PCr and P(i) recovery were delayed in the first minute after stimulation. Because the tension decrease during fatigue is known to be the same in the two genotypes, we conclude that a lack of CA III impairs mitochondrial ATP synthesis.

Lower extremity skeletal muscle function in persons with incomplete spinal cord injury.

STUDY DESIGN: A cross-sectional study design. OBJECTIVES: To characterize and specifically quantify impairments in muscle function after chronic incomplete spinal cord injury (SCI). SETTING: University of Florida, Gainesville, FL, USA. METHODS: Voluntary and electrically elicited contractile measurements were performed and voluntary activation deficits were quantitatively determined in the knee extensor and ankle plantar flexor muscle groups in 10 individuals with chronic incomplete SCI (C5-T8, ASIA C or D) and age-, gender-, height- and body weight matched healthy controls. RESULTS: Persons with incomplete-SCI were able to produce only 36 and 24% of the knee extensor torque and 38 and 26% of the plantar flexor torque generated by noninjured controls in the self-reported less-involved and more-involved limbs, respectively (P<0.05). In addition, both indices of explosive or instantaneous muscle strength, torque200 (absolute torque reached at 200 ms) and the average rate of torque development (ARTD) were dramatically reduced in the ankle plantar flexor and knee extensor muscle groups in persons with incomplete-SCI. However, the deficit in instantaneous muscle strength was most pronounced in the ankle plantar flexor muscles, with an 11.7-fold difference between the torque200 measured in the self-reported more involved limb and a 5-fold difference in the less-involved limb compared to control muscles. Voluntary activation deficits ranged between 42 and 66% in both muscle groups. Interestingly, electrically elicited contractile properties did not differ between the groups. CONCLUSION: The resultant impact of incomplete-SCI is that affected muscles not only become weak, but slow to develop voluntary torque. We speculate that the large deficit in torque200 and ARTD in the ankle plantar flexors muscles of persons with incomplete-SCI may limit locomotor function. The results presented in this study provide a quantitative and sensitive assessment of muscle function upon which future research examining rehabilitation programs aimed at restoring muscle function and promoting functional recovery after incomplete-SCI may be based.

Resistance training improves strength and functional capacity in persons with multiple sclerosis.

The purpose of this study was to evaluate the effect of an eight-week progressive resistance training programme on lower extremity strength, ambulatory function, fatigue and self-reported disability in multiple sclerosis (MS) patients (mean disability score 3.7 +/- 0.8). Eight MS subjects volunteered for twice weekly training sessions. During the first two weeks, subjects completed one set of 8-10 reps at 50% of maximal voluntary contraction (MVC) of knee flexion, knee extension and plantarflexion exercises. In subsequent sessions, the subjects completed one set of 10-15 repetitions at 70% of MVC. The resistance was increased by 2-5% when subjects completed 15 repetitions in consecutive sessions. Isometric strength of the quadriceps, hamstring, plantarflexor and dorsiflexor muscle groups was assessed before and after the training programme using an isokinetic dynamometer. Magnetic resonance images of the thigh were acquired before and after the exercise programme as were walking speed (25-ft), number of steps in 3 min, and self-reported fatigue and disability. Knee extension (7.4%), plantarflexion (52%) and stepping performance (8.7%) increased significantly (P < 0.05). Self-reported fatigue decreased (P < 0.05) and disability tended to decrease (P = 0.07) following the training programme. MS patients are capable of making positive adaptations to resistance training that are associated with improved ambulation and decreased fatigue.

Effects of testosterone replacement therapy on skeletal muscle after spinal cord injury.

STUDY DESIGN: Randomized control. OBJECTIVE: To examine the effects of testosterone replacement therapy (TRT) on skeletal muscle 11 weeks after complete SCI. SETTING: Athens, Georgia USA. METHODS: Soleus (SOL), gastrocnemius (GA), tibialis anterior (TA), vastus lateralis (VL) and triceps brachii (TRI) muscles were taken from twelve young male Charles River rats 11 weeks after complete SCI (T-9 transection, n=8) or sham surgery (n=4). Rats received either TRT (two 5 cm capsules, n=4) or empty capsules (n=8) implanted at surgery. Muscle samples were sectioned and fibers analyzed qualitatively for myosin ATPase and quantitatively for succinate dehydrogenase (SDH), alpha-glycerol-phosphate dehydrogenase (GPDH) and actomyosin ATPase (qATPase) activities using standard techniques. RESULTS: SCI decreased average fiber size (49+/-4%) in affected muscles and the percentage of slow fibers in SOL (93+/-3% to 17+/-2%). In addition, there was a decrease in SDH and an increase in GPDH and qATPase activities across the four hind-limb muscles of the SCI animals. Fiber size in the TRI was increased (31+/-2%) by SCI while enzyme activities were not altered. Average fiber size across the four hind limb muscles was decreased by only 30% in TRT SCI animals and their SOL contained 39+/-2% slow fibers. TRT also attenuated changes in enzyme activities. There was no effect of TRT on the TRI relative to SCI. CONCLUSIONS: TRT was effective in attenuating alterations in myofibrillar proteins during 11 weeks of SCI in affected skelatal muscles. SPONSORSHIP: Supported by a grant from The National Institutes of Health (HD-33738) and HD-37645 to KV, and HD-39676 to GAD.

Specific detection of type III iodothyronine deiodinase protein in chicken cerebellar purkinje cells.

Because iodothyronine deiodinases play a crucial role in the regulation of the available intracellular T(3) concentration, it is important to determine their cellular localization. In brain, the presence of type III iodothyronine deiodinase (D3) seems to be important to maintain homeostasis of T(3) levels. Until now, no cellular localization pattern of the D3 protein was reported in chicken brain. In this study polyclonal antisera were produced against specific peptides corresponding to the D3 amino acid sequence. Their use in immunocytochemistry led to the localization of D3 in the Purkinje cells of the chicken cerebellum. Both preimmune serum as well as the primary antiserum exhausted with the peptide itself were used as negative controls. Extracts of chick cerebellum and liver were made in the presence of Triton X-100 to solubilize the membrane-bound deiodinases. Using these extracts in Western blot analysis, a band of the expected molecular weight ( approximately 30 kDa) could be detected in both tissues. Using a full-length (32)P-labeled type III deiodinase cRNA probe, we identified a single mRNA species in the cerebellum that was of the exact same size as the hepatic control mRNA (+/-2.4 kb). RT-PCR, followed by subcloning and sequence analysis, confirmed the expression of D3 mRNA in the chicken cerebellum. In this study we provide the first evidence of the presence of the D3 protein in a neuronal cell type, namely Purkinje cells, by means of immunocytochemical staining. We were able to detect a protein fragment corresponding to the expected molecular mass (30 kDa) for type III deiodinase by means of Western blot analysis. RT-PCR as well as Northern blot analysis confirmed the presence of D3 mRNA in the cerebellum.

Magnetic resonance spectroscopy of the masseter muscle in different facial morphological patterns.

The aims of this study were (1) to develop a reliable noninvasive method to evaluate the masseter muscle metabolism, by using (31)P-magnetic resonance spectroscopy, and (2) to evaluate the metabolic profile of the masseter muscle in subjects with various facial patterns. The maxillary-mandibular relationship, which varied from hypodivergent to hyperdivergent, was measured on lateral cephalograms of 20 adults, 22 to 35 years of age. (31)P-spectra were acquired from the masseter muscle at rest with a custom-made, single-turn, double-tuned, 3 x 5-cm oblong surface coil. The inorganic phosphate to phosphocreatine (Pi/PCr) ratios were measured and compared in relation to vertical and sagittal cephalometric measurements. A statistically significant (R(2) = 0.65, r = 0.81, P = .001) relationship was found between Pi/PCr ratio and the palatal-to-mandibular plane angle. As the maxillary-to-mandibular divergence increased, the Pi/PCr ratio decreased. This correlation suggests that muscles with a higher Pi/PCr ratio have a higher resting metabolic activity than those with a lower Pi/PCr ratio. Consequently, these muscles may keep bone under more tension and influence its growth in a more horizontal direction. Another possible explanation of the results is that the fiber type composition of the masseter muscle varies with facial morphology.

Metabolic enzymes and phenotypic expression among human locomotor muscles.

Percutaneous biopsies were taken from the right vastus lateralis (VL), tibialis anterior (TA), soleus (Sol), and lateral gastrocnemius (LG) muscles of eight recreationally active adult males. Approximately 60 fibers in each sample were analyzed for their type (I, IIa, or IIx), cross-sectional area (CSA), and succinic dehydrogenase (SDH), alpha glycerol phosphate dehydrogenase (GPDH) and calcium-activated actomyosin adenosine triphosphatase (qATPase) activities. This was done to test the hypothesis that metabolic enzyme activities are more reflective of the functional diversity among human locomotor muscles than fiber type composition. The results showed that enzymatic characteristics differed more or less than expected between muscles of the same or different fiber type. For example, the relative CSA occupied by fast fibers was only about 50% greater in the mixed (LG and VL) than in the slow (Sol and TA) muscles (57 vs. 38%). At the same time, average fiber SDH activity and fiber type specific SDH:qATPase*%CSA, both used as estimates of fatigue resistance, were greater in Sol and LG than in TA and VL. As a result, the two slow muscles and the two mixed muscles had different values, and a mixed muscle (LG) had higher values than a slow muscle (TA). The findings suggest that differences in enzymatic profile, more than fiber type composition, afford human locomotor muscles the capacity to perform their purportedly divergent functional tasks.

Relationship between muscle T2* relaxation properties and metabolic state: a combined localized 31P-spectroscopy and 1H-imaging study.

A multi-volume 31P-magnetic resonance spectroscopy localization procedure was implemented to compare directly muscle metabolism and proton T2* relaxation properties in the human plantar flexor muscles during exercise. Localized 31P-spectra were collected simultaneously from the medial gastrocnemius, lateral gastrocnemius and soleus muscles during exercise using beta1-insensitive Hadamard Spectroscopic Imaging (HSI). 1H T2*-weighted gradient-echo images were acquired at rest and immediately following high-intensity plantar flexion exercise. T2* mapping of the individual calf muscles showed that plantar flexion with the knee extended produces significant increases (P < 0.0001) in the mean (SEM) T2* of the medial [35.6 (1.2) ms vs 28.5 (0.5) ms at rest] and lateral gastrocnemius [35.6 (0.9) ms vs 26.2 (0.9) ms at rest], but not in the soleus [26.7 (0.6) ms vs 27.3 (0.8) ms at rest]. In accordance with the changes in T2*, the ratio of inorganic phosphate to phosphocreatine (Pi:PCr) and the intracellular muscle pH shifted significantly in the gastrocnemii, while the soleus showed no change in muscle pH and only a moderate increase in Pi-to-Ph. Comparison of spectroscopic and relaxation parameters in both gastrocnemius muscles revealed a significant relationship between post-exercise T2* and intracellular pH (r = 0.72-0.76) and Pi-to-Ph ratios (r = 0.81-0.88) during exercise. Using an improved method of localization, this study confirms the existence of a strong relationship between transverse relaxation properties and the metabolic state in skeletal muscles engaged in heavy exercise.

Effects of immobilization on plantar-flexion torque, fatigue resistance, and functional ability following an ankle fracture.

BACKGROUND AND PURPOSE: The goal of this investigation was to study the recovery of ankle plantar-flexor peak torque, fatigue resistance, and functional ability (stair climbing, walking) following cast immobilization in patients with ankle fractures. SUBJECTS: The participants were 10 patients who underwent open reduction-internal fixation and 8 weeks of cast immobilization following a fracture of the ankle mortise and 10 age- and sex-matched, noninjured comparison subjects. METHODS: Plantar-flexor torque and fatigue resistance were measured at 1, 5, and 10 weeks of rehabilitation using an isokinetic dynamometer. Ankle plantar-flexor peak torque and fatigue resistance were correlated to timed ambulation, timed stair climbing, and unilateral heel-rises. RESULTS: Following immobilization, plantar-flexor peak torque was decreased at all angular speeds and positions. The decrease in peak torque was associated with an increase in fatigue resistance. With rehabilitation, ankle plantar-flexor torque and fatigue resistance normalized. Regression analysis revealed a strong relationship between plantar-flexor peak torque and functional measures. By 10 weeks post-immobilization, peak torque, fatigue resistance, and all measures of functional performance had returned to control levels. CONCLUSION AND DISCUSSION: The decrease in muscle performance, functional ability, and fatigue resistance induced by 8 weeks of cast immobilization can be reversed with 10 weeks of supervised physical therapy. In addition, this study demonstrated that ankle-plantar flexor torque is a good predictor of stair-climbing and walking performance in patients with ankle fractures.

In vivo ATP synthesis rates in single human muscles during high intensity exercise.

1. In vivo ATP synthesis rates were measured in the human medial gastrocnemius muscle during high intensity exercise using localized 31P-magnetic resonance spectroscopy (31P-MRS). Six-second localized spectra were acquired during and following a 30 s maximal voluntary rate exercise using a magnetic resonance image-guided spectral localization technique. 2. During 30 s maximal voluntary rate exercise, ATPase fluxes were predominantly met by anaerobic ATP sources. Maximal in vivo glycogenolytic rates of 207 +/- 48 mM ATP min-1 were obtained within 15 s, decreasing to 72 +/- 34 mM ATP min-1 by the end of 30 s. In contrast, aerobic ATP synthesis rates achieved 85 +/- 2 % of their maximal capacity within 9 s and did not change throughout the exercise. The ratio of peak glycolytic ATP synthesis rate to maximal oxidative ATP synthesis was 2.9 +/- 0.9. 3. The non-Pi, non-CO2 buffer capacity was calculated to be 27.0 +/- 6. 2 slykes (millimoles acid added per unit change in pH). At the cessation of exercise, Pi, phosphomonoesters and CO2 were predicted to account for 17.2 +/- 1.5, 5.57 +/- 0.97 and 2.24 +/- 0.34 slykes of the total buffer capacity. 4. Over the approximately linear range of intracellular pH recovery following the post-exercise acidification, pHi recovered at a rate of 0.19 +/- 0.03 pH units min-1. Proton transport capacity was determined to be 16.4 +/- 4.1 mM (pH unit)-1 min-1 and corresponded to a maximal proton efflux rate of 15.3 +/- 2.7 mM min-1. 5. These data support the observation that glycogenolytic and glycolytic rates are elevated in vivo in the presence of elevated Pi levels. The data do not support the hypothesis that glycogenolysis follows Michealis-Menten kinetics with an apparent Km for [Pi] in vivo. 6. In vivo -measured ATP utilization rates and the initial dependence on PCr and glycolysis were similar to those previously reported in in situ studies involving short duration, high intensity exercise. This experimental approach presents a non-invasive, quantitative measure of peak glycolytic rates in human skeletal muscle.

Spectral quantitation by principal component analysis using complex singular value decomposition.

Principal component analysis (PCA) is a powerful method for quantitative analysis of nuclear magnetic resonance spectral data sets. It has the advantage of being model independent, making it well suited for the analysis of spectra with complicated or unknown line shapes. Previous applications of PCA have required that all spectra in a data set be in phase or have implemented iterative methods to analyze spectra that are not perfectly phased. However, improper phasing or imperfect convergence of the iterative methods has resulted in systematic errors in the estimation of peak areas with PCA. Presented here is a modified method of PCA, which utilizes complex singular value decomposition (SVD) to analyze spectral data sets with any amount of variation in spectral phase. The new method is shown to be completely insensitive to spectral phase. In the presence of noise, PCA with complex SVD yields a lower variation in the estimation of peak area than conventional PCA by a factor of approximately 2. The performance of the method is demonstrated with simulated data and in vivo 31P spectra from human skeletal muscle.

Autonomic testing in patients with chronic fatigue syndrome.

The purpose of this study was to determine whether chronic fatigue syndrome (CFS) patients show autonomic dysfunction at the cardiac level and if so, to discover whether these abnormalities explain the fatiguability and/or other symptoms in CFS. The study population consisted of 21 CFS patients (Centers for Disease Control and Prevention [CDC] criteria, 1988) and 13 age- and sex-matched healthy controls. The autonomic testing consisted of: (1) postural challenge: registration of heart rate and blood pressure (BP) and heart rate variability in supine and in upright position (tilted to 70 degrees); (2) Valsalva maneuver; (3) handgrip test; (4) cold pressor test; and (5) heart rate response to deep breathing. Statistical analysis was performed using the Mann Whitney rank sum test; results of the test were considered significant at the 0.05 level. After tilting heart rate was significantly higher in CFS patients compared with healthy controls (mean CFS = 88.9 beats/min vs control = 77.9 beats/min; P <0.01). Low frequency power after tilting was significantly higher in CFS patients compared with controls (mean CFS = 0.603 vs control = 0.428; P = 0.02). There was a trend toward an increased heart rate during the cold pressor test. Other parameters did not differ between the CFS and control populations. The observed changes point toward a sympathetic overactivity in CFS patients when they are exposed to stress. Parasympathetic abnormalities could not be observed. Therefore, our findings provide no real explanation for the fatigue and intolerance to physical exertion in these patients.