Probing motor dynamics at the muscle level-Acoustic myography in Parkinson's disease.

Acoustic myography (AMG) noninvasively probes muscle activity. We explored whether AMG captures abnormal motor activity in patients with Parkinson's disease (PD) and how this activity is modulated by antiparkinsonian medication. Twenty patients with PD underwent AMG of the biceps, triceps, extensor carpi radialis longus, and adductor policis muscles of the more affected arm during active and passive movements, using a mobile AMG device (CURO, Denmark). AMG and assessment of motor symptoms were performed in a pragmatic off-medication state, as well as one and 3 h after oral intake of 200 mg levodopa. Three AMG parameters were calculated using the CURO analysis system. Motor efficiency was expressed by the E-score, muscle fiber recruitment by the temporal T-score, spatial summation by the S-score, and S/T ratio. Twenty age- and sex-matched healthy subjects served as controls. Group mean values were statistically compared using unpaired two-tailed adjusted t-test and ANOVA with Tukey´s correction for multiple comparison (p ≤ 0.05). For the biceps and extensor carpi radialis longus muscles, the active movement S:T ratio was lower in PD relative to healthy controls. The E-score was also lower during active and passive flexion/extension movements in the off-medication state. No significant between-group differences in the AMG scores were noted for the triceps muscle during active or passive movements. The active S:T ratio and the E-score during active elbow flexion and extension may offer a useful means to quickly assess abnormal motor activity and the effect of drug treatment in PD.

Supplementation with camelina oil prevents negative changes in the artery in orchidectomized rats.

The aim of the research was to examine the influence of orchidectomy on the elasticity and wall structure of the abdominal aorta in male rats and to check whether camelina oil treatment has an effect on aorta wall characteristics in orchidectomized rats. Forty 2-month-old male Wistar rats were used in the experiment: 10 animals underwent a sham testis repositioning operation (SHO) and 30 rats were orchidectomized (ORX). After the convalescence period, the SHO and ORX1 rats were given physiological saline intragastrically for 8 weeks; simultaneously, the other rats received camelina oil at the dose of 5 g/kg/b.w. (ORX2) or 9 g/kg/b.w. (ORX3) once a day. At the end of experiment, the animals were euthanized and fragments of the aorta were sampled for elasticity measurement and for histomorphometric and immunohistochemistry analysis. Orchidectomy caused a significant increase in the thickness of the total wall and its particular layers, mean intensity of elastin fluorescence in the tunica intima-media, and the volume of collagen I in tunica adventitia of the abdominal aorta in comparison to the other groups. The mean intensity of collagen I fluorescence in the tunica adventitia and tunica intima-media was significantly lower in the aorta of the orchidectomized rats. The values of the histomorphometric parameters of animals receiving camelina oil were lower than in the ORX1 group and higher than in the SHO rats. The values of the other parameters analyzed after the camelina oil treatment were similar to those in the SHO rats. In conclusion, our study showed that orchidectomy induced changes in the abdominal aorta wall characteristic for aging. Supplementation with camelina oil prevents negative consequences in the vessel wall structure in males with impaired endocrine function.

Physiological parameters of endurance horses pre- compared to post-race, correlated with performance: a two race study from scandinavia.

Few studies have investigated the physiological parameters of endurance horses in Scandinavia. Hence, this two race study has focused on the effects of endurance racing in terms of equine clinicopathological blood parameters, heart score, and fluid use. Race A involved 15 horses (120 km). Two pre- and one post-race blood samples were taken, body condition score was assessed in triplicate pre-race, and an ECG was used to determine heart score. Race B involved 16 horses (65-120 km). One pre- and two post-race blood samples were taken. For both races, horse data as well as fluid intake estimates and cooling water were noted. Race A showed that blood haematocrit, albumin, sodium, and triglycerides increased significantly with endurance racing, whilst chloride, glucose, iron, and potassium decreased significantly. In race B, blood creatinine, cholesterol, and inorganic phosphate continued to increase significantly during the first post-race sampling period compared to pre-race levels, whilst iron, which decreased significantly during the race, increased significantly over the two post-race sampling periods. It is concluded that whilst no correlation between heart score and speed was observed, a significant correlation exists between experience and changes in blood parameters with endurance racing and between fluid intake and average speed.

Non-invasive assessment of equine muscular function: A case study.

Assessment of muscle function after an injury or during recovery is of great importance in the veterinary field. Accelerometry, bioimpedance analysis and mechanomyography/acoustic myography have been used to assess human muscular problems, but have not been applied to the veterinary clinic. We report the clinical use of these techniques in a 12-year-old Danish Warmblood horse presenting with recurring and shifting lameness. Acoustic myography, assessing both the amplitude and frequency of active muscles, was employed to locate the specific area of muscle injury, the right hip, which exhibited minimal fibre recruitment giving rise to considerable weakness. This specific region was assessed by accelerometry which revealed a normal step interval for the injured leg when compared with the contralateral, but a weaker acceleration and strike force. Finally, an assessment of muscle resistance (R) and reactance (Xc) using bioimpedance confirmed a regional loss of muscle mass and a loss of cellular integrity compared with the contralateral limb.

Virtual reflected-light microscopy.

Research on better methods to digitally represent microscopic specimens has increased over recent decades. Opaque specimens, such as microfossils and metallurgic specimens, are often viewed using reflected light microscopy. Existing 3D surface estimation techniques for reflected light microscopy do not model reflectance, restricting the representation to only one illumination condition and making them an imperfect recreation of the experience of using an actual microscope. This paper introduces a virtual reflected-light microscopy (VRLM) system that estimates both shape and reflectance from a set of specimen images. When coupled with anaglyph creation, the system can depict both depth information and illumination cues under any desired lighting configuration. Digital representations are compact and easily viewed in an online setting. A prototype used to construct VRLM representations is comprised only of a microscope, a digital camera, a motorized stage and software. Such a system automatically acquires VRLM representations of large batches of specimens. VRLM representations are then disseminated in an interactive online environment, which allows users to change the virtual light source direction and type. Experiments demonstrate high quality VRLM representations of 500 microfossils.

Vitamin D, surface electromyography and physical function in uraemic patients.

BACKGROUND: Muscle function is impaired in uraemic patients and several causes have been proposed. Deficiency of 25-hydroxyvitamin D (25-OHD), which affects muscle function in non-uraemic patients, may very well also be associated with the myopathy found in these patients. The aim of this study was to investigate the association between 25-OHD and muscle function as well as physical function in chronic kidney disease (CKD) and peritoneal dialysis (PD) patients. METHODS: In this cross-sectional study, 21 adult patients with CKD stage 3-5 and 21 patients treated with PD were included. Standard biochemistry parameters were measured including 25-OHD, 1,25-dihydroxycholecalciferol (1,25-OHD) and parathyroid hormone analysis. Muscle function was determined by 30-second surface electromyography (sEMG) recordings of a right thigh muscle (vastus lateralis) and a second left finger muscle (second dorsal interosseous) under voluntary contractions. Physical function was determined using a 30-second Chair Stand Test and the Short Form 36 quality of life questionnaire. Clinical characteristics were collected from the patient records. RESULTS: Moderate vitamin 25-OHD deficiency (<40 nmol/l) was measured in 52% of patients with CKD and in 71% of the patients on PD. Severe deficiency (<15 nmol/l) was measured in 14% of patients on PD. There were no significant differences between the CKD and PD patients in terms of sEMG results. 25-OHD was not correlated to any results from the tests of sEMG or physical function. However, a higher sEMG frequency and signal root mean square (RMS) were positively associated with a higher Chair Stand Test score. Time to maximum sEMG frequency was negatively correlated to the Chair Stand Test score (p < 0.05), and positively correlated to the level of comorbidity (p < 0.05). sEMG signal peak-peak amplitude, frequency and RMS were positively correlated to the quality of life scales Physical Function, Role Physical, General Health, Vitality, Social Function, Mental Health, and Physical Component Scale (p < 0.001). CONCLUSIONS: 25-OHD deficiency was prevalent in uraemic patients in the present study. Muscle function as determined using sEMG and the Chair Stand Test was not associated with 25-OHD. The results may be biased by the limited variation in 25-OHD and masked by effects of several other variables in this very sick population.

The long-term effect of alpha-ketoglutarate, given early in postnatal life, on both growth and various bone parameters in pigs.

The long-term effect of alpha-ketoglutarate (AKG) given for 21-24 days post-partum, on the skeleton of commercial pigs, was investigated. In experiment A, 12 pigs were given AKG [0.1 g/kg of body weight (b.w.) per day per os], while 12 controls were administered vehicle. At day 169, the left and right femur, humerus and sixth ribs were analysed for mechanical and geometrical properties and quantitative computed tomography. In experiment B, 32 piglets were divided equally into an AKG group (0.3 g/kg of b.w. per day) or a control group. Blood, taken at days 24 and 53 was analysed for plasma 17 beta-oestradiol. The main bone effect of AKG was to increase bone length in the sixth rib (7.3%, p < 0.01), ultimate strength (23%, p < 0.05), Young s modulus (52%, p < 0.001) and maximum elastic strength (31%, p = 0.056) compared with controls. In both experiments, AKG preferentially increased the growth of female piglets, whilst for male piglets AKG had the opposite effect. In addition, AKG elevated plasma 17 beta-oestradiol levels compared to those of controls at the end of the period of treatment (20%, p = 0.002). It is concluded that AKG has long-term effects on rib properties when given early in postnatal life whilst it elevates plasma 17 beta-oestradiol levels only so long as it is being administered.

Biological effects of 2-oxoglutarate with particular emphasis on the regulation of protein, mineral and lipid absorption/metabolism, muscle performance, kidney function, bone formation and cancerogenesis, all viewed from a healthy ageing perspective state of the art--review article.

The fact that men and women are living longer than they have ever done before is something in which we can all rejoice. However, the process of ageing is associated with changes in skeletal, muscular, gastrointestinal, neural hormonal and metabolic processes that seriously affect an individual's performance and quality of life. Indeed, such changes can be contributory to a loss of independence in the elderly. This state-of-the art address highlights the main changes found to occur with ageing whilst simultaneously reporting findings of in vivo and in vitro studies designed to elucidate the potential of the Krebs cycle intermediate - alpha-ketoglutaric acid (AKG) in protecting elderly body systems from failing and degradation. The topics of paramount importance include impaired bone structure and strength, amino acid and mineral absorption, muscle performance, as well as highlighting the role of Krebs cycle intermediates in the debilitating changes that occur with end-stage renal failure and the regulation of the lipid metabolism. Finally, focus will be given to the role of 2-oxoglutarate as a potent protective factor in connection with the development of malignant cells in the body.

De electricitatis catholici musculari - Concerning the electrical properties of muscles, with emphasis on meat quality.

This study aims to explore the potential of evoked non-invasive surface electromyography (SEMG) analysis, in predicting meat quality traits in livestock. Evoked SEMG is a system that records, transdermally, electrical signals generated in muscle fibres upon external stimulation. These signals are reported as compound muscle action potentials (CMAP). CMAP parameters of LD correlated negatively and significantly to ultimate pH (pH 24h) at day 61, but not at day 153 after birth, and a similar albeit positive correlation was observed for muscle glycogen content. Muscle glycogen content and pH 24h correlated negatively in LD and BF. Negative significant correlations between CMAP parameters and shear force were found in LD at day 153 after birth, which might, in the range of the recording electrodes, reflect the combined effect of large cross-sectional area fibres and reduced perimysium content per unit volume of muscle. The fact that correlations between CMAP characteristics and quality traits of both metabolic and non metabolic origin could be established, warrants a fuller investigation of this method in terms of its potential as a predictive tool for meat quality traits in live animals.

Direct measurements of nitric oxide release in relation to expression of endothelial nitric oxide synthase in isolated porcine mitral valves.

The aim of this study was to measure the direct release of nitric oxide (NO) from the porcine mitral valve using a NO microelectrode. Furthermore, the expression and localization of endothelial nitric oxide synthase (eNOS) in the mitral valve was studied using immunohistochemistry, Western blotting and RT-PCR. Results show that bradykinin increases NO release from mitral valves (DeltaBradykinin: 33.71 +/- 10.41 nm NO, P < 0.001, n = 10), whereas N-nitro-l-arginine methyl esther (l-NAME) decreases NO release when compared with basal level (Deltal-NAME: 82.69 +/- 15.66 nm NO, P < 0.005, n = 4). Both protein and mRNA expression of eNOS in mitral valves and in isolated valvular endothelial cells suggest that the NO release is mainly associated with the mitral valve endothelium. It is concluded that direct NO release from porcine mitral valves coincides with eNOS expression. This study documents useful techniques for investigations into the role of local NO release in mitral valve diseases.

The uremic environment and muscle dysfunction in man and rat.

BACKGROUND: Patients reaching end-stage renal disease experience debilitating fatigue, with progression of this disease, rendering patients dysfunctional in their everyday lives. METHODS: In vivo measurements of muscle function, assessed using surface electromyography (EMG), were made on 25 patients prior to and after a session of hemodialysis (HD) treatment, along side in vitro measurements of muscle function in isolated rat muscles incubated in normal or uremic conditions approximating to those found in uremic rats (rat uremic: RU) or uremic humans (human uremic: HU). RESULTS: HD significantly affected plasma values, e.g. reducing urea (69%), creatinine (62%), potassium (23%) and phosphate (48%) concentrations in patients (all p < 0.01). Treatment also improved the EMG frequency of 2nd dorsal interosseous (fast-twitch) (p < 0.01), although no change was noted for vastus lateralis (slow-twitch). In isolated rat muscles, a uremic environment had no significant effect on slow-twitch soleus during field stimulation, however, in fast-twitch extensor digitorum longus, a significant 23% (RU) and 22% (HU) faster rate of decline in force was measured, compared to controls (p < 0.001 and p < 0.01, respectively). CONCLUSION: It is concluded that (1) muscle weakness and its electrophysiological correlates may be rapidly induced by uremic solutes and rapidly reversed when the solutes are removed by dialysis, and (2) fast-twitch muscles are more readily affected by uremic conditions than slow-twitch muscles.

Effects of antisecretory factor-derived peptides on contractions in guinea pig colon.

Two antisecretory factor (AF)-derived peptides have been studied in relation to effects on motility of guinea pig colon. Colon segments were isolated from adult guinea pigs and incubated in Tyrode Ringer. Motility was measured as the force and frequency of contractions upon addition of the derived peptides AF 1 (8 amino acids (aa)) and AF 3 (10 amino acids). At the lowest concentration (5 pM), peptide AF 1 induced a negative effect on the force of contraction in colon segments; an effect that was abolished by the cholinergic agonist carbachol. Peptide AF 3 induced a significant increase in the force of colon contractions at all concentrations (5-180 pM), with carbachol only reducing the effect of peptide AF 3 at a concentration of 15 pM. Both peptides increased contractile frequency, although the overall response was lower for peptide AF 3 than for peptide AF 1. It is concluded that antisecretory factor-derived peptides may play a role in regulating colon motility such that under pathophysiological conditions, they may serve to hasten the evacuation of noxious agents from the large intestine.

Gastrointestinal-tract models and techniques for use in safety pharmacology.

The gastrointestinal tract (GI tract), which extracts nutrients, electrolytes, minerals, and water, is prone to injury as a result of oral drug administration. Clinical assessment of the GI tract is often limited to measurements of transit time and observations of vomiting or diarrhoea, despite the existence of methods and techniques capable of assessing specific changes in GI function at the membrane, cell, and whole animal levels. Membrane studies, record the uptake of solutes, and electrolyte transport, assessing the affects of compounds on transepithelial GI transport and flux. Such methods lend themselves to permeability, immunohistochemistry, morphology, and molecular biology techniques. Isolated cells from the GI tract or cultured cell lines provide knowledge of regulation and function at a cellular level, whilst motility patterns, taken in vivo or from biopsies, provide information at a more integrated level. In anesthetised animals, ligated segments of the intestine can be infused with test compounds, providing information about absorptive and secretory processes important for the treatment of diarrhoea. Computer simulations and modelling are used to predict the disposition of a chemical and its metabolite and can, to some extent, replace animal testing, thereby reducing development costs. Indeed, software programs can be used to simulate the dissolution, absorption, distribution, metabolism, and excretion (ADME) properties of potential drugs in the human GI tract. Finally, advances in the field of imaging, combined with endoscopy, have resulted in a wireless capsule, allowing the inspection of the GI tract anatomy and pathology without surgical intervention. It is concluded that the field of safety pharmacology could rapidly, cheaply, and routinely incorporate membrane, isolated tissue, and endoscopy techniques for GI tract testing of drugs.

Alpha-ketoglutarate treatment early in postnatal life improves bone density in lambs at slaughter.

This study has investigated the long-term effect on skeletal development of a short postnatal period of oral alpha-ketoglutarate (AKG) administration, a compound known to regulate the synthesis of proline, which in turn is important for collagen production. Male lambs born to Shropshire ewes were used in this study. Lambs were randomly assigned to either an AKG-treated group or a control group receiving an equal volume of distilled water. AKG-treated lambs received 0.1 g/kg body weight orally from the first 14 days of postnatal life. Lambs were slaughtered at approximately 130 day of life and a body weight of 43-49 kg. Plasma samples, collected from lambs at days 14 and 130, were analyzed for IGF-1 concentration using sheep-specific RIA kits. Bone development was determined on the femur in terms of geometrical and mechanical properties and quantitative computed tomography (QCT). Trabecular bone density, cortical bone density, and the mechanical properties of the bones were significantly higher for AKG-treated compared with control lambs. However, neither plasma IGF-1 concentration nor the geometrical properties of the bones were significantly influenced by AKG treatment. It is concluded that early postnatal treatment of lambs with AKG positively affects bone strength, an effect that does not appear to be mediated by an increased plasma IGF-1 concentration.

The CATH database: an extended protein family resource for structural and functional genomics.

The CATH database of protein domain structures (http://www.biochem.ucl.ac.uk/bsm/cath_new) currently contains 34 287 domain structures classified into 1383 superfamilies and 3285 sequence families. Each structural family is expanded with domain sequence relatives recruited from GenBank using a variety of efficient sequence search protocols and reliable thresholds. This extended resource, known as the CATH-protein family database (CATH-PFDB) contains a total of 310 000 domain sequences classified into 26 812 sequence families. New sequence search protocols have been designed, based on these intermediate sequence libraries, to allow more regular updating of the classification. Further developments include the adaptation of a recently developed method for rapid structure comparison, based on secondary structure matching, for domain boundary assignment. The philosophy behind CATHEDRAL is the recognition of recurrent folds already classified in CATH. Benchmarking of CATHEDRAL, using manually validated domain assignments, demonstrated that 43% of domains boundaries could be completely automatically assigned. This is an improvement on a previous consensus approach for which only 10-20% of domains could be reliably processed in a completely automated fashion. Since domain boundary assignment is a significant bottleneck in the classification of new structures, CATHEDRAL will also help to increase the frequency of CATH updates.

Effects of endothelin on submandibular salivary responses to parasympathetic stimulation in anaesthetized sheep.

Submandibular responses to stimulation of the parasympathetic chorda tympani nerve have been investigated in anaesthetized sheep before, during and after an intracarotid infusion of endothelin, which reduced the blood flow through the gland by 56+/-5%. Stimulation of the peripheral end of the chorda tympani nerve produced a frequency-dependent increase in the flow of submandibular saliva, and in sodium, potassium and protein output. The reduction in submandibular blood flow, which occurred in response to endothelin, was associated with a decrease in the flow of saliva at all frequencies tested amounting on average to 44+/-6% (P<0.01). The flow of saliva was linearly related to the blood flow before and after endothelin. Both parameters were also linearly related during the infusion of endothelin and the regression lines were parallel. Salivary sodium concentration was significantly increased at the lower frequencies (1 and 2 Hz). Protein output was generally reduced but the difference only achieved statistical significance during stimulation at 1 Hz (P<0.01). Thus, submandibular secretory responses to parasympathetic stimulation are significantly attenuated by reducing the blood flow through the gland in this way.

Alzheimer morphology is not increased in dialysis-associated encephalopathy and long-term hemodialysis.

This study examines the role of aluminium in the etiology of Alzheimer's disease (AD). Brains taken at autopsy (n = 50) from patients with a history of long-term hemodialysis (HD) and intake of aluminium (Al)-containing drugs were examined by light microscopy. Using our modified silver stain we have been able to demonstrate and clearly discriminate between AD changes and dialysis-associated encephalopathy (DAE) on paraffin sections; evaluation was done with a 3-point scale. DAE morphology is characterized by lysosome-derived intracytoplasmic, Al-containing, pathognomonic, argyrophilic inclusions in choroid plexus epithelia, cortical glia and neurons. A statistically significant difference was found between the amounts of drug-related Al ingested and the degree of DAE-related morphological change (P < 0.001). On the other hand no apparent microscopical increase in AD morphology was found. No AD changes were seen whatsoever in patients under the age of 60, despite a history of long-term HD with ingestion of "pure" Al up to 2.5 kg. Patients over 60 years of age occasionally presented with sparse deposits of beta A4 amyloid (beta A4) and/or a low incidence of AD-type neurofibrillary tangles (NFT). In accordance with CERAD criteria these were identified as normal, age-related phenomena (P < 0.001 for beta A4; P < 0.001 for NFT). Rare, isolated cases from a group of 127 long-term hemodialyzed patients have been reported previously, who presented with intermingled, clearly distinguishable lesions of both age-related AD morphology and DAE changes. Comparison of AD morphology with an age-matched control group was not statistically significant (P > 0.6 for beta A4, P > 0.7 for NFT). In our experience, Al does not cause an increase in AD morphology, at least not in terms of bioavailable Al in drugs or as a result of long-term HD.

A rapid classification protocol for the CATH Domain Database to support structural genomics.

In order to support the structural genomic initiatives, both by rapidly classifying newly determined structures and by suggesting suitable targets for structure determination, we have recently developed several new protocols for classifying structures in the CATH domain database (http://www.biochem.ucl.ac.uk/bsm/cath). These aim to increase the speed of classification of new structures using fast algorithms for structure comparison (GRATH) and to improve the sensitivity in recognising distant structural relatives by incorporating sequence information from relatives in the genomes (DomainFinder). In order to ensure the integrity of the database given the expected increase in data, the CATH Protein Family Database (CATH-PFDB), which currently includes 25,320 structural domains and a further 160,000 sequence relatives has now been installed in a relational ORACLE database. This was essential for developing more rigorous validation procedures and for allowing efficient querying of the database, particularly for genome analysis. The associated Dictionary of Homologous Superfamilies [Bray,J.E., Todd,A.E., Pearl,F.M.G., Thornton,J.M. and Orengo,C.A. (2000) Protein Eng., 13, 153-165], which provides multiple structural alignments and functional information to assist in assigning new relatives, has also been expanded recently and now includes information for 903 homologous superfamilies. In order to improve coverage of known structures, preliminary classification levels are now provided for new structures at interim stages in the classification protocol. Since a large proportion of new structures can be rapidly classified using profile-based sequence analysis [e.g. PSI-BLAST: Altschul,S.F., Madden,T.L., Schaffer,A.A., Zhang,J., Zhang,Z., Miller,W. and Lipman,D.J. (1997) Nucleic Acids Res., 25, 3389-3402], this provides preliminary classification for easily recognisable homologues, which in the latest release of CATH (version 1.7) represented nearly three-quarters of the non-identical structures.

Assigning genomic sequences to CATH.

We report the latest release (version 1.6) of the CATH protein domains database (http://www.biochem.ucl. ac.uk/bsm/cath ). This is a hierarchical classification of 18 577 domains into evolutionary families and structural groupings. We have identified 1028 homo-logous superfamilies in which the proteins have both structural, and sequence or functional similarity. These can be further clustered into 672 fold groups and 35 distinct architectures. Recent developments of the database include the generation of 3D templates for recognising structural relatives in each fold group, which has led to significant improvements in the speed and accuracy of updating the database and also means that less manual validation is required. We also report the establishment of the CATH-PFDB (Protein Family Database), which associates 1D sequences with the 3D homologous superfamilies. Sequences showing identifiable homology to entries in CATH have been extracted from GenBank using PSI-BLAST. A CATH-PSIBLAST server has been established, which allows you to scan a new sequence against the database. The CATH Dictionary of Homologous Superfamilies (DHS), which contains validated multiple structural alignments annotated with consensus functional information for evolutionary protein superfamilies, has been updated to include annotations associated with sequence relatives identified in GenBank. The DHS is a powerful tool for considering the variation of functional properties within a given CATH superfamily and in deciding what functional properties may be reliably inherited by a newly identified relative.

Measurement of force and both surface and deep M wave properties in isolated rat soleus muscles.

In isolated soleus muscles of 4-wk-old rats, M wave parameters were recorded with surface and deep recording electrodes and examined in relation to both twitch and tetanic force. Addition of ouabain (10(-5) M for 16 min) to isolated muscles caused an approximately 40% decrease in twitch amplitude and area (P < 0.01) that was associated with a 98% decrease in surface M wave amplitude, a 78% decrease in deep M wave amplitude (both P < 0.001), a 98% decrease in surface M wave area (P < 0.01), 48% of which occurred within 60 s of addition of ouabain (P < 0.05), and a 55% decrease in deep M wave area (P < 0.05). The decrease in twitch parameters on addition of ouabain was most closely correlated with deep M wave area (r = 0.92). Direct tetanic stimulation at a frequency of 30 Hz resulted in an initial potentiation of M waves, which was not seen at a frequency of 90 Hz. Instead, 90 Hz stimulation resulted in a prompt decrease in tetanic force that was correlated with a decrease in both deep M wave amplitude (r = 0.94; P < 0.01) and deep M wave area (r = 0.96; P < 0.01). It is concluded that simultaneous surface and deep recordings involving area and amplitude are fundamental to analysis of the effects of pharmacological agents on muscle performance and the use of M waves as predictors of muscle excitability.