Fine-wire recordings of flexor hallucis brevis motor units up to maximal voluntary contraction reveal a flexible, nonrigid mechanism for force control.

Our current knowledge on the neurophysiological properties of intrinsic foot muscles is limited, especially at high forces. This study therefore aimed to investigate the discharge characteristics of single motor units in an intrinsic foot muscle, namely flexor hallucis brevis, during voluntary contractions up to 100% of maximal voluntary contraction. We measured the recruitment threshold and discharge rate of flexor hallucis brevis motor units using indwelling fine-wire electrodes. Ten participants followed a target ramp up to maximal voluntary contraction by applying a metatarso-phalangeal flexion torque. We observed motor unit recruitment thresholds across a wide range of isometric forces (ranging from 10 to 98% of maximal voluntary contraction) as well as across a wide range of discharge rates (ranging from 4.8 to 23.3 Hz for initial discharge rate and 9.5 to 34.2 Hz for peak discharge rate). We further observed patterns of high variability in recruitment threshold and discharge rate as well as crossover in discharge rate between motor units within the same participant. These findings suggest that the force output of a muscle is generated through a mechanism with substantial variability rather than relying on a rigid organization, which is in contrast to the proposed onion-skin theory. The demands placed on the plantar intrinsic foot muscles during high- and low-force tasks may explain these observed neurophysiological properties.NEW & NOTEWORTHY We recorded for the first time single motor unit action potential trains in the flexor hallucis brevis, a short toe muscle, over the full range of maximum voluntary contraction. Its motor units are recruited up to very high (98%) recruitment thresholds with a substantial range of discharge rates. We further show high variability with crossover of discharge rates as a function of recruitment threshold both between participants and between motor units within participants.

A 5-year longitudinal study of association of maximum bite force with development of frailty in community-dwelling older adults.

To determine whether maximum bite force (MBF), an objective measure of oral function, is associated with development of frailty in community-dwelling older adults. This prospective cohort study included community-dwelling Japanese adults aged 75 years at baseline (n = 322). Baseline MBF was measured using an electronic recording device (Occlusal Force-Meter GM10). Follow-up examinations, including physical fitness and anthropometric evaluation and structured questionnaires, were administered annually over a 5-year period to determine the incidence of frailty, which was defined by the presence of 3 or more of the following 5 components derived from the Cardiovascular Health Study: low level of mobility, low physical activity level, weakness, shrinking and poor endurance and energy. Adjusted hazard ratios (HRs) of incidence of frailty according to sex-stratified tertiles of baseline MBF were calculated using Cox proportional hazards regression models. During the follow-up, 49 participants (15.2%) developed frailty. Participants in the lower tertile of MBF exhibited a significantly greater risk of frailty than those in the upper tertile. After adjustment for sex, depression, diabetes and Eichner index, the adjusted HRs for frailty in the upper through lower tertiles of MBF were 1.00 (reference), 1.27 (95% confidence interval [CI]: 0.50-3.20) and 2.78 (95% CI: 1.15-6.72), respectively (P for trend = .01). Poor oral function, as indicated by low MBF, increases the risk of development of frailty among elderly men and women.

Effect of short-term maximal voluntary co-contraction training on neuromuscular function.

The present study aimed to examine the effect of short-term training utilizing voluntary co-contraction with maximal efforts. 23 healthy young men (training group: TG, n = 13; control group: CG, n = 10) participated in this study. TG conducted a 4-week training program (3 days/week), which consisted of 4 s simultaneous maximal voluntary contractions of elbow flexors and extensors at 90° of the elbow joint, followed by 4 s muscle relaxation (10 repetitions/set, 5 sets/day). Before and after the intervention, maximal voluntary isometric and isokinetic torques and the muscle thicknesses of the elbow flexors and extensors were determined. The electromyograms (EMGs) of the 2 muscle groups during isometric maximal voluntary contraction (MVC) were also recorded. After the intervention, CG did not show any significant changes in all measured variables. In TG, significant increases were found in the agonist EMG activities during MVC, and maximal isometric and isokinetic torques of the elbow flexors and extensors, without significant changes in the muscle thicknesses and involuntary coactivation levels during MVC. The current results indicate that the training mode with maximal voluntary co-contraction is effective for improving the force-generating capabilities of the exercising muscles, without any increases in the level of involuntary coactivation during MVC.

Speeding up of sedimentation under confinement.

We show an increase of the sedimentation velocity as small particles are confined in circular capillaries. In general, confinement slows down sedimentation. But, we show that at low Reynolds numbers and in 1D confinement this is not the case. Particle sedimentation velocity is not homogeneous, which can lead to the formation of structures. These structures are enhanced and stabilized in the presence of walls and in the absence of other dissipative mechanisms. As a consequence, it is possible to achieve sedimentation velocities that even exceed the Stokes velocity. The segregation at critical capillary diameters has been directly observed using a large scale model. These simple experiments offer a new insight into the old problem of sedimentation under confinement.

Overexpression of cIAP2 contributes to 5-FU resistance and a poor prognosis in oral squamous cell carcinoma.

BACKGROUND: Resistance to 5-fluorouracil (5-FU) is a major obstacle in treating oral squamous cell carcinoma (OSCC). However, little is known about apoptosis resistance, which contributes to 5-FU resistance in OSCC. METHODS: We focussed on the cellular inhibitor of apoptosis protein 2 (cIAP2) on the basis of a DNA microarray data using parental and 5-FU-resistant OSCC cell lines. The effects of cIAP2 downregulation on 5-FU sensitivity and apoptosis were evaluated. An immunohistochemical analysis of cIAP2 and related proteins, cIAP1 and X-linked IAP, was performed in 54 OSCC patients who were treated with 5-FU-based chemoradiotherapy and surgery. RESULTS: The downregulation of cIAP2 significantly enhanced the sensitivity of the 5-FU-resistant cells to 5-FU, with a significant increase in apoptosis. The immunohistochemical analysis demonstrated a high cIAP2 tumour expression to significantly correlate with the pathological response to chemoradiotherapy. Furthermore, a Cox regression analysis revealed the cIAP2 expression status (hazard ratio, 4.91; P=0.037) and the pathological response to chemoradiotherapy (hazard ratio, 0.418; P=0.016) to be significant prognostic factors for OSCC patients. CONCLUSION: These novel findings demonstrate that cIAP2 may represent a potentially useful therapeutic target for improving the treatment and survival of OSCC patients, particularly in the setting of 5-FU resistance.

Body mass-based exercise in middle-aged and older women.

The present study examined whether the degree to which muscle strength is improved by a body mass-based home exercise program in middle-aged and older women depends on the force-generating capabilities of the muscles prior to the intervention. 75 women (53-76 years) voluntarily participated in a circuit training program consisting of 5 exercises (16 repetitions/exercise, 2 or 3 circuits/day) using only body mass as resistance for 3 months. The subjects performed the training program 6 days a week in their own home and once a week in a local gym. Before and after intervention, isometric torques during maximal voluntary knee extension (KET) and plantar flexion (PFT) were determined and expressed relative to body mass (KET/BM and PFT/BM, respectively). KET/BM and PFT/BM increased significantly after intervention, and their relative changes were negatively correlated to the absolute values before intervention. Most of the subjects whose KET/BM and PFT/BM values before intervention were greater than 2.8 Nm/kg and 1.7 Nm/kg, respectively, did not show increases in strength after intervention. Thus, although body mass-based exercise at home is effective in improving lower limb strength in middle-aged and older women, the magnitude of the improvement is influenced by the force-generating capability before intervention.

Apical enrichment of human EGF precursor in Madin-Darby canine kidney cells involves preferential basolateral ectodomain cleavage sensitive to a metalloprotease inhibitor.

EGF precursor (proEGF) is a member of the family of membrane-anchored EGF-like growth factors that bind with high affinity to the epidermal growth factor receptor (EGFR). In contrast to human transforming growth factor-alpha precursor (proTGFalpha), which is sorted basolaterally in Madin-Darby canine kidney (MDCK) cells (Dempsey, P., and R. Coffey, 1994. J. Biol. Chem. 269:16878-16889), we now demonstrate that human proEGF overexpressed in MDCK cells is found predominantly at the apical membrane domain under steady-state conditions. Nascent proEGF (185 kD) is not sorted but is delivered equally to the apical and basolateral membranes, where it is proteolytically cleaved within its ectodomain to release a soluble 170-kD EGF form into the medium. Unlike the fate of TGFalpha in MDCK cells, the soluble 170-kD EGF species accumulates in the medium, does not interact with the EGFR, and is not processed to the mature 6-kD peptide. We show that the rate of ectodomain cleavage of 185-kD proEGF is fourfold greater at the basolateral surface than at the apical surface and is sensitive to a metalloprotease inhibitor, batimastat. Batimastat dramatically inhibited the release of soluble 170-kD EGF into the apical and basal medium by 7 and 60%, respectively, and caused a concordant increase in the expression of 185-kD proEGF at the apical and basolateral cell surfaces of 150 and 280%, respectively. We propose that preferential ectodomain cleavage at the basolateral surface contributes to apical domain localization of 185-kD proEGF in MDCK cells, and this provides a novel mechanism to achieve a polarized distribution of cell surface membrane proteins under steady-state conditions. In addition, differences in disposition of EGF and TGFalpha in polarized epithelial cells offer a new conceptual framework to consider the actions of these polypeptide growth factors.

Physical function is weakly associated with angiotensin-converting enzyme gene I/D polymorphism in elderly Japanese subjects.

BACKGROUND: The turning point in the deterioration of physical function seems to occur between the ages of 70 and 80 years. In particular, muscle strength may decline even more in subjects older than 75. A recent study found that the angiotensin-converting enzyme (ACE) genotype also affects physiological left ventricular hypertrophy. A very limited number of papers have examined genetic differences in resistance and endurance forms of a single sporting discipline. OBJECTIVE: The purpose of this study was to evaluate the relationship between ACE genotype and physical function by controlling the known confounding factors including dental status. METHODS: We selected 431 subjects who were aged 76 years and did not require special care for their daily activities. We conducted a medical examination, followed by 5 physical function tests, as follows: (1) maximum hand grip strength, (2) maximal isometric knee extensor strength, (3) maximal stepping rate for 10 s, (4) one-leg standing time with eyes open and (5) 10-meter maximum walking speed. Subjects were genotyped for the ACE intron 16 Alu insertion. In addition, serum concentrations of total cholesterol, total protein, IgA and IgG were measured at a commercial laboratory. The Eichner index was used as an indicator of occlusal condition. Multiple linear regression analysis was performed to evaluate the relationship between the ACE gene insertion/deletion (I/D) polymorphism and physical function considering confounding factors. RESULTS: The ACE gene I/D polymorphism was positively associated with hand grip strength and 10-meter maximum walking speed. Betas of hand grip strength were 0.09 for I/D (p = 0.022) and 0.12 for insertion/insertion (I/I; p = 0.004). Betas of 10-meter walking speed were -0.11 for I/D (p = 0.093) and -0.14 for I/I (p = 0.039). Dental status such as Eichner index class C was significantly associated with one-leg standing time with eyes open (beta -0.11; p = 0.028). CONCLUSION: This study suggests that there is a significant relationship between ACE genotype and physical function. In particular, subjects with the ACE deletion/deletion genotype were associated with upper extremities.

Physical fitness and cognitive function in an 85-year-old community-dwelling population.

BACKGROUND: Little is known about the association between physical fitness and cognitive function in very elderly people (over 80 years of age). OBJECTIVES: To evaluate that relationship in 85-year-old community-dwelling individuals. METHODS: Out of 207 participants (90 males, 117 females) who were 85 years old and community-dwelling, 205 completed the Mini-Mental State Examination (MMSE) for evaluating cognitive function. The numbers of subjects who completed physical fitness measurements such as hand-grip strength, isometric leg extensor strength, one-leg standing time, stepping rate, and walking speed were 198, 159, 169, 168, and 151, respectively. RESULTS: There were significant associations in MMSE with hand-grip strength (right or left hand), isometric leg extensor strength, stepping rate, and walking speed by simple regression analysis. MMSE was still significantly associated with hand-grip strength (beta = 0.305, p = 0.005 for right side; beta = 0.309, p = 0.004 for left side), stepping rate (beta = 0.183, p = 0.046), and walking speed (beta = -0.222, p = 0.014) by multiple regression analysis after adjustments for the amount of education, gender, smoking, drinking, complication of stroke, body weight, body height, regular medical care, serum albumin, blood HbA1c, and marital status. By logistic regression analysis, the prevalence of a normal MMSE score (MMSE >or=24) was increased by 9% with each 1-kg increase in hand-grip strength of the left hand (OR 1.087, 95% CI 1.003-1.179, p = 0.042), and was increased by 6% with each step per 10 s in stepping rate (OR 1.060, 95% CI 1.000-1.122, p = 0.048). CONCLUSION: In a very elderly population of 85-year-olds, cognitive function was associated with some physical fitness measurements, independent of confounding factors.

Surface tension and elasticity of gel studied with laser-induced surface-deformation spectroscopy.

The laser manipulation technique was effectively used for agarose solutions and the frequency specstrum of the surface response to the periodical laser irradiation yielded shear elasticity G and surface tension sigma in the gel. The laser spot size, from 60 mum to 200 mum in radius, was chosen so that either the Rayleigh waves or the capillary waves, selectively excited, associated with G or sigma , respectively. The result of G showed a dependence on the agarose concentration that is consistent with the theoretical prediction of the percolation model, while sigma has little dependence on the concentration. The surface state of 0.2 wt.% agarose solution was controlled with sodium-dodecyl-sulfate (SDS) additives, and sigma of the gel and the sol was observed at different SDS concentrations: The result showed (i) sigma decreased with increasing SDS concentration up to 39 x 10-3 mol/l and kept a constant value thereafter, and (ii) the gel and the sol have the same value of sigma and the same dependence on the concentration. These results were considered from a viewpoint of surface pressure and a partially quantitative discussion was made on the surface adsorbed with SDS and agarose molecules.

Identification of a family of insulin-like growth factor II secreted by cultured rat epithelial-like cell line 18,54-SF: application of a monoclonal antibody.

Somatomedin/insulin-like growth factor (IGF)-like polypeptides (designated SMP) were purified from the serum-free conditioned medium of cultured rat epithelial-like cells, 18,54-SF. A monoclonal antibody (MAb) was produced against partially purified SMP. The antibody was immunoglobulin G1 relatively specific for multiplication-stimulating activity III-2 (rat IGF-II), with a Kd value of 5.6 X 10(-9) M. The antibody showed 100% cross-reactivity with human IGF-II and 10% cross-reactivity with human IGF-I, but did not cross-react with insulin. For purification of SMP, therefore, immunoaffinity chromatography on Sepharose coupled with the MAb was used besides a procedure including ion exchange chromatography, gel filtration, and reverse phase HPLC. The purified SMP (at least five polypeptides) each gave a single peak on reverse phase HPLC and appeared as a single band with an apparent mol wt of 5000-8000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The major components of SMP (designated HP1-SMP and HP3-SMP), which were purified about 100-fold from conditioned medium, stimulated DNA synthesis in human fibroblasts in culture and sulfation in chick embryonic cartilage in culture. These polypeptides showed almost the same cross-reactivity as multiplication-stimulating activity III-2 on RIA with the MAb. The partial amino acid sequences of HP1- and HP3-SMP were determined, and these polypeptides were identified with rat IGF-II.

cDNA cloning and chromosomal mapping of genes encoding novel protein kinases termed PKU-alpha and PKU-beta, which have nuclear localization signal.

We have cloned cDNAs for novel serine/threonine protein kinases (PK), termed PKU-alpha and PKU-beta, by screening a bacteriophage expression library for kinase activity. Sequence analysis of PKU-alpha and PKU-beta genes revealed that their open reading frames (ORF) were 2151 and 2361 nucleotides (nt) encoding polypeptides of 717 and 787 amino acid (aa) residues, respectively. The deduced aa sequences of PKU-alpha and PKU-beta contained typical serine/threonine PK domains at the C-terminal region and were 86% identical to each other, indicating that they belong to the same PK family. Northern analysis reveals that they are expressed in nearly all human tissues and in cultured cells. The genes for PKU-alpha and PKU-beta were mapped to chromosome 17q23 and 8p12-p22, respectively, by fluorescence in situ hybridization. The proteins encoded by both cDNAs contain a putative nuclear localization signal (NLS) in their N-terminal region. These signals are likely to function in nuclear localization. Glutathione S-transferase (GST)-fusions to regions of PKU-alpha and beta containing the NLS were efficiently localized to the nucleus. In addition, PKU-beta transiently expressed in COS-1 cells was predominantly nuclear. PKU-alpha and PKU-beta differ: a consensus sequence for a nt binding motif is present near the NLS of PKU-beta. These results suggest that PKU-alpha and beta may phosphorylate serine and/or threonine residues on similar proteins, but their activities are regulated through distinct interactions with a nuclear component.

Correlations of the expression of fibroblast growth factor-2, vascular endothelial growth factor, and their receptors with angiogenesis in synovial tissues from patients with internal derangement of the temporomandibular joint.

Synovitis in internal derangement of the temporomandibular joint (TMJ) is accompanied by the growth of new blood vessels. Fibroblast growth factor-2 (FGF-2) and vascular endothelial growth factor (VEGF) are well-characterized angiogenic factors. The objective of this study was to elucidate the correlation between the expression of FGF-2, VEGF, and their receptors-FGF receptor-1 (FGFR-1) and VEGF receptor-1 (Flt-1)-with microvessel density in synovial tissues of the TMJ. Using an immunohistochemical technique, we examined 47 joints (45 patients) with internal derangement. Individual microvessel density was evaluated by means of the CD34 antibody, a specific endothelial marker. The correlation between the percentage of immuno-positive cells and microvessel density was evaluated. In multiple logistic regression analysis, the correlation between the percentage of Flt-1-positive cells and microvessel density was significant [p = 0.005, odds ratio = 1.071, 95% confidence interval = 1.021-1.124]. These results suggest that the expression of the VEGF/Flt-1 system is involved in angiogenesis in inflamed synovial tissue in the TMJ.

Different antitumor activities of anti-bFGF neutralizing antibodies: heparin-binding domain provides an inefficient epitope for neutralization in vivo.

Basic fibroblast growth factor (bFGF) plays an important role in tumor growth and angiogenesis. To elucidate the efficient recognition sites by anti-bFGF neutralizing antibodies, we generated two anti-bFGF neutralizing monoclonal IgG1 antibodies (mAbs), 2G11 and 1E6, recognizing different sites, and estimated as binding to the heparin-binding and the receptor-binding regions of bFGF, respectively, both of which have been shown to be important for its receptor interaction. Despite their high in vitro anti-bFGF activity in the absence of heparin, 2G11, with in vitro activity in competition with heparin, failed to inhibit the in vivo tumor growth of bFGF-producing RPMI4788 cells, though 1E6, showing non-competition with heparin, exhibited a significant antitumor effect. These results show that the heparin-binding domain of bFGF provides an inefficient epitope for in vivo neutralization of anti-bFGF mAb, and anti-bFGF neutralizing mAbs without competition against heparin have the potential to show in vivo antitumor effects.

HLA antigens as immunogenetic markers of alcoholism and alcoholic liver disease.

HLA antigens were studied among 94 chronic alcoholics. Concerning A and B-loci, there was no significant change of phenotype frequency (PF) in the HLA typing between the patients and controls (80 healthy subjects). However, there was a significant difference in the PF of CW3 between chronic alcoholics and controls (58.5% in alcoholics vs 30.0% in controls). The corrected p value was less than 0.05 with relative risk value being 3.29 HLA-DR loci were also detected in 26 patients, but there was no significant difference between the patients and controls. All alcoholics were subdivided according to the hepatic morphology, and the PF of HLA was examined. A significant high frequency of HLA CW3 was found in patients with hepatitis (64%) compared to controls (30%). There was also an increased PF of CW3 in the liver cirrhosis group (59% in cirrhosis group vs 30% in controls). In conclusion, chronic alcoholics have a significantly higher PF of HLA-CW3 as compared to controls. This characteristic feature becomes even more distinct in alcoholics with severe hepatic lesions.

[Serum-free culture of tumor cells--recent problems in autocrine hypothesis].

Tumor cell lines in culture that can grow in protein-free medium are suitable for studying the mechanism of autonomous growth of tumor cells. We have studied the mechanism of autonomous growth of three cell lines which were established from human solid tumors. These cell lines produce basic fibroblast growth factor (bFGF), but it does not seem to act as an autocrine growth factor. bFGF produced by these cells is accumulated in the nuclei and may act as an autointrafactor for autonomous growth of these cells.

1998 ISEK Congress Keynote Lecture: The use of electromyography in applied physiology. International Society of Electrophysiology and Kinesiology.

Electromyogram (EMG) analyses (surface, intramuscular and evoked potentials) in studies of muscle function have attracted increasing attention during recent years and have been applied to assess muscle endurance capacity, anaerobic and lactate thresholds, muscle biomechanics, motor learning, neuromuscular relaxation, optimal walking and pedalling speeds, muscle soreness, neuromuscular diseases, motor unit (MU) activities (MU recruitment and rate coding), and skeletal muscle fatigue. This paper deals with the use of EMG analyses employed in the area of applied physiology and is divided into three sections: surface EMG analyses; intramuscular EMG analyses; and evoked potential analyses.

The muscle sound properties of different muscle fiber types during voluntary and electrically induced contractions.

Soundmyogram (SMG) and electromyogram signals were recorded simultaneously from the relatively fast medial gastrocnemius (MG) and slow soleus (SOL) during voluntary and electrically induced contractions. Using a spike-triggered averaging technique, the averaged elementary sound and corresponding MU spikes were also obtained from about 35 different MUs identified. The rms-SMG of MG increased as a function of force (P < 0.01). On the contrary, these values for SOL increased up to 60% MVC (P < 0.01), but decreased at 80% MVC. The relationship between the peak to peak amplitude of SMG and MU spike indicated significant positive correlations (r = 0.631 to approximately 0.657, P < 0.01). During electrical stimulation at 5 Hz, the SMG power spectral peak frequency (PF) was matched with stimulation frequency in both muscles. At higher stimulation frequencies, e.g., > 15 Hz, only in the MG was SMG-PF synchronized with stimulation frequency; the slow SOL did not show such synchronization. Our data suggest that the SMG frequency components might reflect active motor unit firing rates, and that the SMG amplitude depends upon mechanical properties of contraction, muscle fiber composition, and firing rate during voluntary and electrically induced contractions.

Localization of basic fibroblast growth factor (bFGF) in a metastatic cell line (AT-3) established from the Dunning prostatic carcinoma of rat: application of a specific monoclonal antibody.

Localization of basic fibroblast growth factor (bFGF) in a metastatic cell line, AT-3, established from the Dunning prostatic carcinoma of rat was determined by two immunological techniques using a specific monoclonal antibody against bFGF. Concentration of bFGF in cell extract was measured by sandwich radioimmunoassay (RIA) with heparin-Sepharose and 125I-labeled monoclonal antibody. bFGF concentration in the extract of AT-3 cells increased with increasing concentration of NaCl in extraction buffer. Localization of bFGF in AT-3 cells was determined by counting radioactivity of 125I-labeled monoclonal antibody binding to AT-3 cells before or after increasing permeability of the cells. The binding increased significantly by this treatment, indicating that bFGF within the cells was detected.

The expression of fibroblast growth factor-2 and fibroblast growth factor receptor-1 in chondrocytes in synovial chondromatosis of the temporomandibular joint. report of two cases.

Synovial chondromatosis (SC) is a rare, benign condition characterized by the formation of metaplastic cartilaginous nodules. The expression of fibroblast growth factor-2 (FGF-2) and fibroblast growth factor receptor-1 (FGFR-1) in two cases of SC of the temporomandibular joint (TMJ) were immunohistochemically studied. The possible roles of FGF-2 and FGFR-1 in SC of the TMJ are discussed.