Vincent J. Caiozzo

Accurate prediction of VO2max in cycle ergometry.

Numerous equations exist for predicting VO2max from the duration (an analog of maximal work rate, Wmax) of a treadmill graded exercise test (GXT). Since a similar equation for cycle ergometry (CE) was not available, we saw the need to develop such an equation, hypothesizing that CE VO2max could be accurately predicted due to its more direct relationship with W. Thus, healthy, sedentary males (N = 115) and females (N = 116), aged 20-70 yr, were given a 15 W.min-1 CE GXT. The following multiple linear regression equations which predict VO2max (ml.min-1) from the independent variables of Wmax (W), body weight (kg), and age (yr) were derived from our subjects: Males: Y = 10.51 (W) + 6.35 (kg) - 10.49 (yr) + 519.3 ml.min-1; R = 0.939, SEE = 212 ml.min-1. Females: Y = 9.39 (W) + 7.7 (kg) - 5.88 (yr) + 136.7 ml.min-1; R = 0.932, SEE = 147 ml.min-1 Using the 95% confidence limits as examples of worst case errors, our equations predict VO2max to within 10% of its true value. Internal (double cross-validation) and external cross-validation analyses yielded r values ranging between 0.920 and 0.950 for the male and female regression equations. These results indicate that use of the equations generated in this study for a 15 W.min-1 CE GXT provides accurate estimates of VO2max.

Isokinetic torque imbalances in the rotator cuff of the elite water polo player

The specific repetitive activity of water polo, like base ball pitching, emphasizes adduction and internal rota tion. This study used the Cybex II to evaluate the isokinetic strength of the rotator cuff in elite water polo players and in a group of control subjects. The water polo players were significantly stronger than the con trols. Of greater importance was the confirmation of imbalances in the rotator cuff force couples of adduc tion/abduction and external/internal rotation. These changes are similar to those reported for pitchers. The adductors in the water polo group had gained in relative strength resulting in an increase in the adduction/ab duction ratio to about 2:1. The internal rotators had gained in relative strength resulting in a decrease in the external/internal ratio to about 0.6:1. For both force couples the differences are more apparent at a slow speed. Side-to-side differences were not significant.

Plasticity of skeletal muscle phenotype: mechanical consequences.

Muscles are complex biological machines that perform a wide variety of mechanical activities. Over the past 30 to 40 years, a large amount of effort has been devoted to understanding cellular/molecular responses of skeletal muscle to various altered physiological states (e.g., altered loading state induced via immobilization/spaceflight, resistance training). Many cellular/molecular adaptations brought about by such interventions act on underlying processes that regulate activation, force and velocity of shortening/lengthening, and relaxation. In this context, measurements of mechanical properties (e.g., force–velocity relationship) are important, because they can provide insight into the physiological consequences of such adaptations. During the course of the past 10 to 15 years, a number of investigators have employed the work‐loop technique to provide a more realistic approach toward understanding muscle function. Additionally, the work‐loop technique provides a unique conceptual perspective that integrates: (1) the length–tension relationship, (2) activation kinetics, (3) the force–velocity relationship in the shortening domain, (4) relaxation kinetics, (5) the force–velocity relationship in the lengthening domain, and (6) the compliance of the passive elastic elements. A discussion of those factors (i.e., factors 2–5) that appear to be highly malleable forms the basis of this paper.

VCP associated inclusion body myopathy and paget disease of bone knock-in mouse model exhibits tissue pathology typical of human disease.

Dominant mutations in the valosin containing protein (VCP) gene cause inclusion body myopathy associated with Pagets disease of bone and frontotemporal dementia (IBMPFD). We have generated a knock-in mouse model with the common R155H mutation. Mice demonstrate progressive muscle weakness starting approximately at the age of 6 months. Histology of mutant muscle showed progressive vacuolization of myofibrils and centrally located nuclei, and immunostaining shows progressive cytoplasmic accumulation of TDP-43 and ubiquitin-positive inclusion bodies in quadriceps myofibrils and brain. Increased LC3-II staining of muscle sections representing increased number of autophagosomes suggested impaired autophagy. Increased apoptosis was demonstrated by elevated caspase-3 activity and increased TUNEL-positive nuclei. X-ray microtomography (uCT) images show radiolucency of distal femurs and proximal tibiae in knock-in mice and uCT morphometrics shows decreased trabecular pattern and increased cortical wall thickness. Bone histology and bone marrow derived macrophage cultures in these mice revealed increased osteoclastogenesis observed by TRAP staining suggestive of Paget bone disease. The VCPR155H/+ knock-in mice replicate the muscle, bone and brain pathology of inclusion body myopathy, thus representing a useful model for preclinical studies.

Myosin isoform expression and MAFbx mRNA levels in hibernating golden-mantled ground squirrels (Spermophilus lateralis).

Hibernating mammals present many unexplored opportunities for the study of muscle biology. The hindlimb muscles of a small rodent hibernator (Spermophilus lateralis) atrophy slightly during months of torpor, representing a reduction in the disuse atrophy commonly seen in other mammalian models. How torpor affects contractile protein expression is unclear; therefore, we examined the myosin heavy‐chain (MHC) isoform profile of ground squirrel skeletal muscle before and after hibernation. Immunoblotting was performed first to identify the MHC isoforms expressed in this species. Relative percentages of MHC isoforms in individual muscles were then measured using SDS‐PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis). The soleus and diaphragm did not display differences in isoforms following hibernation, but we found minor fast‐to‐slow isoform shifts in MHC protein in the gastrocnemius and plantaris. These subtle changes are contrary to those predicted by other models of inactivity but may reflect the requirement for shivering thermogenesis during arousals from torpor. We also measured mRNA expression of the Muscle Atrophy F‐box (MAFbx), a ubiquitin ligase important in proteasome‐mediated proteolysis. Expression was elevated in the hibernating gastrocnemius and the plantaris but was not associated with atrophy. Skeletal muscle from hibernators displays unusual plasticity, which may be a combined result of the intense activity during arousals and the reduction of metabolism during torpor.

Prediction of normal values for lactate threshold estimated by gas exchange in men and women

Abstract Lactate threshold (LT) is an index of exercise capacity and can be estimated from the gas exchange consequences of a metabolic acidosis (LTGE). In recent years, it has emerged as a diagnostic tool in the evaluation of subjects with exercise limitation. The purpose of this study was to develop LTGE prediction equations on a relatively large sample of adults and to cross-validate each equation. A total of 204 healthy, sedentary, nonsmoking subjects (103 men and 101 women), aged 20–70 years, underwent graded exercise testing on a cycle ergometer. The V-slope technique was used to detect LTGE as the oxygen uptake (V˙O2) at the breakpoint of the carbon dioxide output versus V˙O2 relationship. Multiple linear regression was used to develop 12 equations with combinations of the following predictor variables: age, height, body mass, and fat-free mass. Eight of the equations are gender-specific and four are generalized with gender as a dummy variable. The equations were cross-validated using the predicted residual sum of squares (PRESS) method. The results demonstrate that the equations had relatively high multiple correlations (0.577–0.863) and low standard errors of the estimate (0.123–0.228 1 · min−1). The PRESS method demonstrated that the equations are generalizable, i.e., can be used in future studies without a significant loss of accuracy. Since we tested only healthy, sedentary subjects, our equations can be used to predict the lower limit of normal for a given subject. Using individual data for healthy and diseased subjects from the literature, we found that our gender-specific equations rarely miscategorized subjects unless they were obese and mass was a predictor variable. We conclude that our equations provide accurate predictions of normal values for LTGE and that they are generalizable to other subject populations.

Artificial gravity as a countermeasure to microgravity: a pilot study examining the effects on knee extensor and plantar flexor muscle groups

The goal of this project was to examine the effects of artificial gravity (AG) on skeletal muscle strength and key anabolic/catabolic markers known to regulate muscle mass. Two groups of subjects were selected for study: 1) a 21 day-bed rest (BR) group (n = 7) and 2) an AG group (n = 8), which was subjected to 21 days of 6 degrees head-down tilt bed rest plus daily 1-h exposures to AG (2.5 G at the feet). Centrifugation was produced using a short-arm centrifuge with the foot plate approximately 220 cm from the center of rotation. The torque-velocity relationships of the knee extensors and plantar flexors of the ankle were determined pre- and posttreatment. Muscle biopsy samples obtained from the vastus lateralis and soleus muscles were used for a series of gene expression analyses (mRNA abundance) of key factors implicated in the anabolic vs. catabolic state of the muscle. Post/pre torque-velocity determinations revealed greater decrements in knee extensor performance in the BR vs. AG group (P < 0.04). The plantar flexors of the AG subjects actually demonstrated a net gain in the torque-velocity relationship, whereas in the BR group, the responses declined (AG vs. BR, P < 0.001). Muscle fiber cross-sectional area decreased by approximately 20% in the BR group, whereas no losses were evident in the AG group. RT-PCR analyses of muscle biopsy specimens demonstrated that markers of growth and cytoskeletal integrity were higher in the AG group, whereas catabolic markers were elevated in the BR group. Importantly, these patterns were seen in both muscles. We conclude that paradigms of AG have the potential to maintain the functional, biochemical, and structural homeostasis of skeletal muscle in the face of chronic unloading.

Pathogenic accumulation of APP in fast twitch muscle of IBM patients and a transgenic model

Inclusion body myositis (IBM) is the most common age-related degenerative skeletal muscle disorder. The aberrant intracellular accumulation of the beta-amyloid (Abeta) peptide within skeletal muscle is a pathological hallmark of IBM. Skeletal muscle is comprised of both slow and fast twitch fibers, which are present in different proportions in various muscles. It remains unclear if fast and/or slow twitch fibers are differentially involved in IBM pathogenesis. To better understand the molecular pathogenesis of IBM, we analyzed human IBM muscle biopsies and muscle from a transgenic mouse model of IBM (MCK-betaAPP). Here we report that the majority of histopathologically-affected fibers in human IBM biopsies were type II fast fibers. Skeletal muscle from MCK-betaAPP mice exhibited higher transgene expression and steady-state levels of human betaAPP in fast type IIB fibers compared to slow type I fibers. These findings indicate that fast twitch fibers may selectively accumulate and be more vulnerable to betaAPP- and Abeta-mediated damage in IBM. These findings also highlight parallels between the MCK-betaAPP mice and the human IBM condition.

Effects of distraction on muscle length: mechanisms involved in sarcomerogenesis.

Although a great deal of interest has been given to understanding the mechanisms involved in regulating the radial growth that occurs because of resistance training, much less has been given to studying the longitudinal growth of skeletal muscle that occurs because of passive stretch. The current authors provide a brief overview of key issues relevant to the longitudinal growth of skeletal muscle that occurs during distraction osteogenesis. Specifically, five key issues are addressed: (1) the pattern of sarcomerogenesis during distraction; (2) sarcomerogenesis and altered expression of sarcomeric and nonsarcomeric genes; (3) the satellite cell hypothesis; (4) mitogenic factors; and (5) new approaches for studying the longitudinal growth of skeletal muscle. A discussion is provided that revolves around the concept of a negative feedback loop. One of the most interesting issues to be resolved in muscle biology is the role of satellite cells in regulating the growth of skeletal muscle. Currently, it is not known whether satellite cell activation is a prerequisite for the longitudinal growth of skeletal muscle. Gene chip analyses provide a paradoxical view, showing that distraction osteogenesis results in the upregulation of a gene, GADD45, involved with growth arrest and deoxyribonucleic acid destruction.

A new concept in laryngeal muscle: multiple myosin isoform types in single muscle fibers of the lateral cricoarytenoid.

This report describes the first known investigation of canine laryngeal muscle in which single fibers were dissected and their myosin heavy chain (MHC) isoform content was analyzed. Both SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot techniques were used. The data from single fiber SDS-PAGE indicate that the lateral cricoarytenoid (LCA) is predominantly a fast muscle composed of the following MHC isoforms: Type I, 16.3%; Type IIA, 71.3%; Type IIX, 10.4%; and Type IIB, 2.0%. The results reveal a phenomenon that, to our knowledge, has not been previously described for laryngeal muscle: the presence of two or more MHC isoforms in a single canine LCA muscle fiber. A large number (41%) of muscle fibers coexpressed two or more MHC isoforms. The three most common patterns of coexpression were Type IIA/IIX (72%), Type IIA/I (16%), and Type IIA/IIX/I (8%). Interestingly, the fast Type IIX MHC isoform was typically present with other isoforms and rarely found by itself in individual fibers. Additional experiments are underway to determine whether other laryngeal muscles exhibit such an unusually high ratio of MHC isoform polymorphism.