Muscles A Medical Dictionary Bibliography and Annotated Research Guide to Internet References

MUSCLES A MEDICAL DICTIONARY, BIBLIOGRAPHY, AND ANNOTATED RESEARCH GUIDE TO INTERNET REFERENCES JAMES N. PARKER, M.D. AND PHILIP M. PARKER, PH.D., EDITORS

ii ICON Health Publications ICON Group International, Inc. 4370 La Jolla Village Drive, 4th Floor San Diego, CA 92122 USA Copyright 2004 by ICON Group International, Inc. Copyright 2004 by ICON Group International, Inc. All rights reserved. This book is protected by copyright. No part of it may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, or otherwise, without written permission from the publisher. Printed in the United States of America. Last digit indicates print number: 10 9 8 7 6 4 5 3 2 1 Publisher, Health Care: Philip Parker, Ph.D. Editor(s): James Parker, M.D., Philip Parker, Ph.D. Publisher's note: The ideas, procedures, and suggestions contained in this book are not intended for the diagnosis or treatment of a health problem. As new medical or scientific information becomes available from academic and clinical research, recommended treatments and drug therapies may undergo changes. The authors, editors, and publisher have attempted to make the information in this book up to date and accurate in accord with accepted standards at the time of publication. The authors, editors, and publisher are not responsible for errors or omissions or for consequences from application of the book, and make no warranty, expressed or implied, in regard to the contents of this book. Any practice described in this book should be applied by the reader in accordance with professional standards of care used in regard to the unique circumstances that may apply in each situation. The reader is advised to always check product information (package inserts) for changes and new information regarding dosage and contraindications before prescribing any drug or pharmacological product. Caution is especially urged when using new or infrequently ordered drugs, herbal remedies, vitamins and supplements, alternative therapies, complementary therapies and medicines, and integrative medical treatments. Cataloging-in-Publication Data Parker, James N., 1961- Parker, Philip M., 1960- Muscles: A Medical Dictionary, Bibliography, and Annotated Research Guide to Internet References / James N. Parker and Philip M. Parker, editors p. cm. Includes bibliographical references, glossary, and index. ISBN: 0-497-11073-3 1. Muscles-Popular works. I. Title.

iii Disclaimer This publication is not intended to be used for the diagnosis or treatment of a health problem. It is sold with the understanding that the publisher, editors, and authors are not engaging in the rendering of medical, psychological, financial, legal, or other professional services. References to any entity, product, service, or source of information that may be contained in this publication should not be considered an endorsement, either direct or implied, by the publisher, editors, or authors. ICON Group International, Inc., the editors, and the authors are not responsible for the content of any Web pages or publications referenced in this publication. Copyright Notice If a physician wishes to copy limited passages from this book for patient use, this right is automatically granted without written permission from ICON Group International, Inc. (ICON Group). However, all of ICON Group publications have copyrights. With exception to the above, copying our publications in whole or in part, for whatever reason, is a violation of copyright laws and can lead to penalties and fines. Should you want to copy tables, graphs, or other materials, please contact us to request permission (E-mail: [email protected]). ICON Group often grants permission for very limited reproduction of our publications for internal use, press releases, and academic research. Such reproduction requires confirmed permission from ICON Group International, Inc. The disclaimer above must accompany all reproductions, in whole or in part, of this book.

iv Acknowledgements The collective knowledge generated from academic and applied research summarized in various references has been critical in the creation of this book which is best viewed as a comprehensive compilation and collection of information prepared by various official agencies which produce publications on muscles. Books in this series draw from various agencies and institutions associated with the United States Department of Health and Human Services, and in particular, the Office of the Secretary of Health and Human Services (OS), the Administration for Children and Families (ACF), the Administration on Aging (AOA), the Agency for Healthcare Research and Quality (AHRQ), the Agency for Toxic Substances and Disease Registry (ATSDR), the Centers for Disease Control and Prevention (CDC), the Food and Drug Administration (FDA), the Healthcare Financing Administration (HCFA), the Health Resources and Services Administration (HRSA), the Indian Health Service (IHS), the institutions of the National Institutes of Health (NIH), the Program Support Center (PSC), and the Substance Abuse and Mental Health Services Administration (SAMHSA). In addition to these sources, information gathered from the National Library of Medicine, the United States Patent Office, the European Union, and their related organizations has been invaluable in the creation of this book. Some of the work represented was financially supported by the Research and Development Committee at INSEAD. This support is gratefully acknowledged. Finally, special thanks are owed to Tiffany Freeman for her excellent editorial support.

v About the Editors James N. Parker, M.D. Dr. James N. Parker received his Bachelor of Science degree in Psychobiology from the University of California, Riverside and his M.D. from the University of California, San Diego. In addition to authoring numerous research publications, he has lectured at various academic institutions. Dr. Parker is the medical editor for health books by ICON Health Publications. Philip M. Parker, Ph.D. Philip M. Parker is the Eli Lilly Chair Professor of Innovation, Business and Society at INSEAD (Fontainebleau, France and Singapore). Dr. Parker has also been Professor at the University of California, San Diego and has taught courses at Harvard University, the Hong Kong University of Science and Technology, the Massachusetts Institute of Technology, Stanford University, and UCLA. Dr. Parker is the associate editor for ICON Health Publications.

vi About ICON Health Publications To discover more about ICON Health Publications, simply check with your preferred online booksellers, including Barnes&Noble.com and Amazon.com which currently carry all of our titles. Or, feel free to contact us directly for bulk purchases or institutional discounts: ICON Group International, Inc. 4370 La Jolla Village Drive, Fourth Floor San Diego, CA 92122 USA Fax: 858-546-4341 Web site: www.icongrouponline.com/health

vii Table of Contents FORWARD .......................................................................................................................................... 1 CHAPTER 1. STUDIES ON MUSCLES................................................................................................... 3 Overview........................................................................................................................................ 3 The Combined Health Information Database................................................................................. 3 Federally Funded Research on Muscles ......................................................................................... 7 E-Journals: PubMed Central ....................................................................................................... 66 The National Library of Medicine: PubMed ................................................................................ 87 Academic Periodicals covering Muscles .................................................................................... 132 Dissertations on Muscles........................................................................................................... 132 CHAPTER 2. NUTRITION AND MUSCLES ....................................................................................... 145 Overview.................................................................................................................................... 145 Finding Nutrition Studies on Muscles...................................................................................... 145 Federal Resources on Nutrition ................................................................................................. 153 Additional Web Resources ......................................................................................................... 154 CHAPTER 3. ALTERNATIVE MEDICINE AND MUSCLES................................................................. 163 Overview.................................................................................................................................... 163 The Combined Health Information Database............................................................................. 163 National Center for Complementary and Alternative Medicine................................................ 164 Additional Web Resources ......................................................................................................... 164 General References ..................................................................................................................... 204 CHAPTER 4. PATENTS ON MUSCLES ............................................................................................. 205 Overview.................................................................................................................................... 205 Patents on Muscles .................................................................................................................... 205 Patent Applications on Muscles ................................................................................................ 240 Keeping Current ........................................................................................................................ 270 CHAPTER 5. BOOKS ON MUSCLES ................................................................................................. 271 Overview.................................................................................................................................... 271 Book Summaries: Federal Agencies............................................................................................ 271 Book Summaries: Online Booksellers......................................................................................... 276 Chapters on Muscles .................................................................................................................. 276 CHAPTER 6. MULTIMEDIA ON MUSCLES ...................................................................................... 281 Overview.................................................................................................................................... 281 Video Recordings ....................................................................................................................... 281 Audio Recordings....................................................................................................................... 287 CHAPTER 7. RESEARCHING MEDICATIONS .................................................................................. 289 Overview.................................................................................................................................... 289 U.S. Pharmacopeia..................................................................................................................... 289 Commercial Databases ............................................................................................................... 298 Researching Orphan Drugs ....................................................................................................... 299 APPENDIX A. PHYSICIAN RESOURCES .......................................................................................... 303 Overview.................................................................................................................................... 303 NIH Guidelines.......................................................................................................................... 303 NIH Databases........................................................................................................................... 305 Other Commercial Databases..................................................................................................... 307 The Genome Project and Muscles .............................................................................................. 307 APPENDIX B. PATIENT RESOURCES............................................................................................... 313 Overview.................................................................................................................................... 313 Patient Guideline Sources.......................................................................................................... 313 News Services and Press Releases.............................................................................................. 322 Newsletter Articles .................................................................................................................... 323 Finding Associations.................................................................................................................. 327

viii Contents APPENDIX C. FINDING MEDICAL LIBRARIES ................................................................................ 329 Overview.................................................................................................................................... 329 Preparation................................................................................................................................. 329 Finding a Local Medical Library................................................................................................ 329 Medical Libraries in the U.S. and Canada ................................................................................. 329 ONLINE GLOSSARIES................................................................................................................ 335 Online Dictionary Directories ................................................................................................... 340 MUSCLES DICTIONARY............................................................................................................ 341 INDEX .............................................................................................................................................. 435

1 FORWARD In March 2001, the National Institutes of Health issued the following warning: \"The number of Web sites offering health-related resources grows every day. Many sites provide valuable information, while others may have information that is unreliable or misleading.\"1 Furthermore, because of the rapid increase in Internet-based information, many hours can be wasted searching, selecting, and printing. Since only the smallest fraction of information dealing with muscles is indexed in search engines, such as www.google.com or others, a non-systematic approach to Internet research can be not only time consuming, but also incomplete. This book was created for medical professionals, students, and members of the general public who want to know as much as possible about muscles, using the most advanced research tools available and spending the least amount of time doing so. In addition to offering a structured and comprehensive bibliography, the pages that follow will tell you where and how to find reliable information covering virtually all topics related to muscles, from the essentials to the most advanced areas of research. Public, academic, government, and peer-reviewed research studies are emphasized. Various abstracts are reproduced to give you some of the latest official information available to date on muscles. Abundant guidance is given on how to obtain free-of-charge primary research results via the Internet. While this book focuses on the field of medicine, when some sources provide access to non-medical information relating to muscles, these are noted in the text. E-book and electronic versions of this book are fully interactive with each of the Internet sites mentioned (clicking on a hyperlink automatically opens your browser to the site indicated). If you are using the hard copy version of this book, you can access a cited Web site by typing the provided Web address directly into your Internet browser. You may find it useful to refer to synonyms or related terms when accessing these Internet databases. NOTE: At the time of publication, the Web addresses were functional. However, some links may fail due to URL address changes, which is a common occurrence on the Internet. For readers unfamiliar with the Internet, detailed instructions are offered on how to access electronic resources. For readers unfamiliar with medical terminology, a comprehensive glossary is provided. For readers without access to Internet resources, a directory of medical libraries, that have or can locate references cited here, is given. We hope these resources will prove useful to the widest possible audience seeking information on muscles. The Editors 1 From the NIH, National Cancer Institute (NCI): http://www.cancer.gov/cancerinfo/ten-things-to-know.



3 CHAPTER 1. STUDIES ON MUSCLES Overview In this chapter, we will show you how to locate peer-reviewed references and studies on muscles. The Combined Health Information Database The Combined Health Information Database summarizes studies across numerous federal agencies. To limit your investigation to research studies and muscles, you will need to use the advanced search options. First, go to http://chid.nih.gov/index.html. From there, select the “Detailed Search” option (or go directly to that page with the following hyperlink: http://chid.nih.gov/detail/detail.html). The trick in extracting studies is found in the drop boxes at the bottom of the search page where “You may refine your search by.” Select the dates and language you prefer, and the format option “Journal Article.” At the top of the search form, select the number of records you would like to see (we recommend 100) and check the box to display “whole records.” We recommend that you type “muscles” (or synonyms) into the “For these words:” box. Consider using the option “anywhere in record” to make your search as broad as possible. If you want to limit the search to only a particular field, such as the title of the journal, then select this option in the “Search in these fields” drop box. The following is what you can expect from this type of search: • Characterization of Esophageal Striated Muscle in Patients with Achalasia Source: Digestive Diseases and Sciences. 45(2): 285-288. February 2000. Summary: Many studies have been conducted analyzing the manometric properties of patients with achalasia, but the striated portion of the esophagus has never been analyzed and is often overlooked. This article reports on a retrospective review of 120 manometric tracings (20 achalasia, 100 controls) performed between 1994 and 1997. Tracings were excluded from patients with chronic cough and nutcracker esophagus. The data were assessed for age, sex, symptoms, duration of symptoms, lower esophageal sphincter (LES) pressure, gastroesophageal gradient, upper esophageal sphincter pressure, smooth muscle contraction amplitude and duration, striated muscle contraction amplitude and duration, length from upper esophageal sphincter to maximal striated muscle contraction, and esophageal length. The maximum striated

4 Muscles muscle contraction amplitude was significantly decreased in achalasia patients with a median amplitude of 45 mm Hg versus 76 mm Hg in the control group. Although the wave forms were similar, the maximum striated muscle contraction duration and the distance from the upper esophageal sphincter in achalasia patients was not significantly different from controls. The length of the esophagus was significantly longer in achalasia patients than in the control group. The authors conclude that patients with achalasia have significantly lower maximum striated muscle contraction amplitudes and longer esophagi, but the duration of the contractions and the configuration of the wave forms are not different. 2 figures. 2 tables. 12 references. • Significance of Pelvic Floor Muscles in Anal Incontinence Source: Gastroenterology. 123(5): 1441-1450. November 2002. Contact: Available from W.B. Saunders Company. 6277 Sea Harbor Drive, Orlando, FL 32887-4800. (800) 654-2452. Website: www.gastrojournal.org. Summary: The pathophysiology of anal incontinence may be elusive using current measurements. This article reports on a study undertaken to establish the role of the levator ani muscles in anal incontinence. The study included 53 patients with anal incontinence, 30 with constipation as disease controls, and 15 healthy controls. The authors evaluated incontinence severity by a 0-12 scale, anorectal function by standard manometric tests, and levator ani contraction by a perineal dynamometer. Patients with incontinence exhibited various physiological abnormalities, but analysis showed that levator ani contraction was the independent variable with strongest relation to the severity of incontinence. Furthermore, in contrast to other physiological parameters, clinical improvement in response to treatment was associated with a marked and significant strengthening of levator ani contraction. The authors conclude by reiterating the importance of levator ani failure in understanding the etiology (cause) of anal incontinence and in predicting response to treatment. 5 figures. 3 tables. 34 references. • Anatomic Plane of Separation Between External Anal Sphincter and Puborectalis Muscle: Clinical Implications Source: Diseases of the Colon and Rectum. 42(3): 374-379. March 1999. Contact: Available from Williams and Wilkins. 352 West Camden Street, Baltimore, MD 21201-2436. Summary: The possible existence of an anatomic and functional separation between the external sphincter and the puborectalis muscle has been reported in the medical literature. In this article, the authors confirm, by means of anatomic and clinical observations, the presence of such a separation and focus on its importance in understanding the pathway of diffusion for some suppurative anal lesions and in planning advanced procedures to spare the sphincter. Twenty adult anatomic specimens of the anal region (from 12 women, and 8 men) were cut in the sagittal, coronal, and paracoronal planes, stained with hematoxylin and eosin, and examined. The pelvic floor musculature was examined in 3 patients undergoing postanal repair operations. Primary posterior and posterolateral anal fistulas in 11 women and 19 men were carefully traced during and after staged fistulotomy. An attempted was made peranally to separate the external sphincter from the puborectalis muscle to spare the sphincter in 4 patients (3 women) aged 56 to 65 years with rectal cancers 4 to 5 cm from the anal verge. The results of these investigations showed a connective plane of separation between the puborectalis muscle and the external sphincter. An anatomicofunctional separation between the puborectalis muscle and the external sphincter was easily

Studies 5 demonstrated during anal repair operations. The authors conclude that the presence of this plane is important to help understand the diffusion of some suppurative anal lesions and to plan advanced procedures to spare the sphincter. 9 figures. 26 references. (AA-M). • Effects of Adductor Muscle Stimulation on Speech in Abductor Spasmodic Dysphonia Source: Laryngoscope. 110(11): 1943-1949. November 2000. Contact: Available from Lippincott Williams and Wilkins. 12107 Insurance Way, Hagerstown, MD 21740. (800) 638-3030 or (301) 714-2300. Fax (301) 824-7390. Summary: This article reports on a study undertaken to determine whether adductor laryngeal muscle stimulation might be a beneficial treatment alternative for abductor spasmodic dysphonia (ABSD). Baseline comparisons were made of measures of voiceless consonant and syllable duration between patients with ABSD (n = 10, two men and eight women, aged 36 to 69 years) and normal control subjects. Speech and voice production with and without muscle stimulation were compared within 10 patients with ABSD. Baseline group comparisons were conducted on measures of syllable and voiceless consonant duration between the patients and the control subjects. Neuromuscular stimulation was applied to the thyroarytenoid or lateral cricoarytenoid muscles in the patients during extended phonation, and measures were made of fundamental frequency and sound pressure level in the stimulated and nonstimulated conditions. Voiceless consonant duration was compared with and without adductor laryngeal muscle stimulation during syllable repetitions and sentences in the patients. Before stimulation, the patients had increased syllable durations in comparison with control subjects. Repeated within patient comparisons with and without stimulation demonstrated significant reductions in voiceless consonant durations during syllable repetition. The more severely affected patients had the greatest reductions in voiceless consonant duration during sentence production. The authors conclude that adductor muscle stimulation improved speech production in patients with ABSD, and the improvement was greatest in the most severely affected patients. Therefore, adductor muscle stimulation has potential for benefiting patients with ABSD. 8 figures. 2 tables. 22 references. • Diabetic Muscle Infarction: An Underdiagnosed Complication of Long-Standing Diabetes Source: Diabetes Care. 26(1): 211-215. January 2003. Contact: Available from American Diabetes Association. 1701 North Beauregard Street, Alexandria, VA 22311. (800) 232-3472. Website: www.diabetes.org. Summary: This article reports on a systematic review of all the reported cases of diabetic muscle infarction (DMI) and its pathogenesis, clinical features, prognostic implications, and management. The authors searched databases (MEDLINE and EMBASE) from their inception to August 2001 and reviewed bibliographies in reports retrieved. A total of 47 references were retrieved; 115 patients and 166 episodes were included. Clinical presentation of DMI is uniform, with acute onset of painful swelling of the affected muscle. DMI was more frequent in women. Of the cases, 59 percent had type 1 diabetes; the mean duration of disease was 14.3 years; and multiple diabetic end-organ complications were noted. DMI affects the lower limbs with abrupt onset of pain and local swelling. Diagnosis is made by biopsy, but the characteristic features in magnetic

6 Muscles resonance imaging (MRI) are very typical. Treatment includes bed rest and administration of analgesics, but recurrence is common. 3 figures. 1 table. 49 references. • Basic Clinical Management of Muscle Strains and Tears Source: Journal of Musculoskeletal Medicine. 20(6): 303-307. June 2003. Summary: This journal article discusses prevention, diagnosis, and treatment of muscle strains and tears. Muscle strains and tears are injuries to a muscle or tendon that occur when the muscle is stretched excessively. Muscle strains are more common than tears. These injuries, especially hamstring strain, are common in athletes. Most patients with muscle strain injuries present after an acute onset of pain during activity. Physical examination may reveal local swelling or ecchymosis; palpitation usually reveals localized tenderness over the myotendinous junction. Radiographs may only show soft tissue swelling but should be obtained if there is any concern about fracture. MRIs may be helpful if the diagnosis is unclear. Acute management of muscle strain injuries includes rest, ice, compression, elevation cryotherapy, and NSAIDs. Prolonged immobilization after strain injury should be avoided, and an exercise program should be instituted after pain and swelling subside to recover range of motion, strength, endurance, and eventually, normal athletic skills. Surgery may be required in the rare case of a complete tear with significant retraction of the muscle from the tendon. Flexible, strong, and warmed-up muscles are the key to strain rehabilitation and injury prevention. 11 references and 2 figures. (AAM). • Muscle Tone Abnormalities Source: Rehabilitation Nursing. 22(3):118-123,130; May/June 1997. Summary: This journal article for health professionals presents current information about alterations in muscle tone. Rehabilitation nurses frequently encounter clients with neurological disorders that adversely affect muscle tone, so, if they understand the physiological etiology of abnormal muscle tone, they can design nursing interventions for various care settings. Topics discussed include the basis of motor control; the relationship between the type of muscle tone alteration and the location of neurological damage; the differences between spasticity, rigidity, and flaccidity; the medical and physical treatment approaches to muscle tone problems; and the nurse's role in managing alterations in muscle tone. 20 references, 2 figures, and 3 tables. (AA-M). • The Effects of Muscle Fatigue on Neuromuscular Function and Anterior Tibial Translation in Healthy Knees Source: American Journal of Sports Medicine. 24(5):615-621. 1996. Summary: This journal article for physicians investigated the effect of quadriceps and hamstring muscle fatigue on anterior tibial translation and muscle reaction time in 6 men and 4 women, all healthy, with an average age of 21.3 years with no known pathologic knee conditions. Each patient underwent a knee examination, arthrometer measurements of tibial translation, subjective functional assessment, and an anterior tibial translation stress test before and after quadriceps and hamstring muscle-fatiguing exercise. The recruitment order of the lower extremity muscles in response to anterior tibial translation did not change with muscle fatigue. However, results showed that an average increase of 32.5 percent in anterior tibial translation (range, 11.4 percent to 85.2 percent) after fatigue. Muscle responses in the gastrocnemius, hamstring, and quadriceps originating at the spinal cord and cortical level showed significant slowing and, in some cases, an absence of activity after the quadriceps and hamstring muscles

Studies 7 were fatigued. The authors indicate that increases in displacement after fatigue strongly correlated (0.62 to 0.96) with a delay in cortical activity (intermediate and voluntary). Muscle fatigue, which appears to affect the dynamic stability of the knee, alters the neuromuscular response to anterior tibial translation. It is suggested that fatigue may play an important role in the pathomechanics of knee injuries in physically demanding sports. 3 tables, 1 figure, and 26 references. (AA) • How To Evaluate the Patient Who Has Muscle Disease Source: Journal of Musculoskeletal Medicine. 17(7): 407-410,413. July 2000. Summary: This journal article provides health professionals with information on the evaluation of a patient who complains of muscle weakness, pain, dysfunction, or fatigue. Many conditions can affect the skeletal muscles, including inflammatory diseases such as polymyositis or one of the connective tissue disorders; endocrine, genetic, and metabolic disorders; and central and peripheral nervous system diseases. The patient history can help establish the severity of the symptoms. When questioning patients about their symptoms, the physician should list the tasks they now have difficulty performing and also quantify as much as possible their degree of weakness or strength. An additional area of inquiry that is crucial for the diagnosis of muscle disease is the family history. Facts gained from the family history may be of great value in suggesting the presence, or at least the strong possibility, of a particular disease. The physical examination should attempt to quantify the patient's muscle strength, assess other aspects of muscle function, and detect abnormal movements and reflex changes. During the examination, the physician should look for abnormal movements and reflex changes, as well as changes in muscle volume. Biochemical analyses of muscle enzymes and serum myoglobin, electromyographic studies, magnetic resonance imaging, sonography, and muscle biopsy can help confirm the diagnosis. 3 tables. (AA-M). Federally Funded Research on Muscles The U.S. Government supports a variety of research studies relating to muscles. These studies are tracked by the Office of Extramural Research at the National Institutes of Health.2 CRISP (Computerized Retrieval of Information on Scientific Projects) is a searchable database of federally funded biomedical research projects conducted at universities, hospitals, and other institutions. Search the CRISP Web site at http://crisp.cit.nih.gov/crisp/crisp_query.generate_screen. You will have the option to perform targeted searches by various criteria, including geography, date, and topics related to muscles. For most of the studies, the agencies reporting into CRISP provide summaries or abstracts. As opposed to clinical trial research using patients, many federally funded studies use animals or simulated models to explore muscles. The following is typical of the type of information found when searching the CRISP database for muscles: 2 Healthcare projects are funded by the National Institutes of Health (NIH), Substance Abuse and Mental Health Services (SAMHSA), Health Resources and Services Administration (HRSA), Food and Drug Administration (FDA), Centers for Disease Control and Prevention (CDCP), Agency for Healthcare Research and Quality (AHRQ), and Office of Assistant Secretary of Health (OASH).

8 Muscles • Project Title: A RODENT MODEL FOR LOCOMOTOR TRAINING WITH FNS Principal Investigator & Institution: Jung, Ranu; Associate Professor; Ctr for Biomedical Engineering; University of Kentucky 109 Kinkead Hall Lexington, Ky 40506 Timing: Fiscal Year 2002; Project Start 17-JAN-2002; Project End 31-OCT-2002 Summary: The long-term goal of this work is to develop strategies for using functional neuromuscular stimulation (FNS) of paralyzed muscles to enhance the recovery of individuals with incomplete spinal cord injury. The proposed work is motivated by three important developments. First, recent basic science and clinical studies have demonstrated that the degree of functional recovery of the injured spinal cord depends on the activity patterns of neural inputs to the spinal cord. Second, recent advances have produced adaptive controllers for FNS systems that provide a means of automatically adjusting stimulation parameters to reliably achieve specified rhythmic movements. Third, rodent models of spinal cord injury (complete and incomplete lesions) are extensively being used at the molecular, cellular, and systems level to investigate the effects of traumatic injury and to assess the results of therapeutic intervention. A combination therapy that utilizes locomotor training with FNS and pharmacological intervention is likely to be the most effective in enhancing the reorganization (plasticity) of the spinal circuitry that is spared after spinal trauma. A rodent model for FNS- assisted locomotion would facilitate quantitative evaluation of therapeutic regimens that include FNS and would provide the ability to characterize effects of FNS-assisted locomotion on the neuroanatomy and neurophysiology of the injured spinal cord. This biomedical engineering research grant proposal will develop a rodent model of locomotor training that utilizes treadmill walking and functional neuromuscular stimulation (FNS) with fixed-pattern and adaptive controllers. Kinematic and electromyogram (EMG) patterns of intact animals will be examined and then used to develop stimulation patterns for FNS-assisted movement. A series of tasks will be performed using FNS stimulation of hindlimb muscles in spinalized rats. These tasks will progress in difficulty from controlling suspended hindlimb movements to controlling hindlimb movements during treadmill locomotion in spinalized rats with partial weight support. Two different FNS control strategies will be used for each movement: a fixed-pattern, or open-loop, stimulation pattern and an adaptive stimulation control system. The adaptive stimulation control system will build upon our previous work and is expected to provide movement patterns that are more accurate and more repeatable. Successful completion of the proposed project will result in a novel animal model for FNS-assisted locomotor training and provide quantitative methods for evaluating locomotor behavior. In future studies, we plan to use a rodent model of incomplete spinal cord injury with FNS-assisted locomotion to test the hypothesis that FNS-assisted locomotor training enhances motor recovery after incomplete spinal cord injury. We anticipate that the improved performance provided by the adaptive control system may enhance the therapeutic effects of the technique. This locomotor training could also be combined with pharmacological intervention, tissue transplant, and neural repair therapies to determine if locomotor training can enhance the effectiveness of these therapies. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: ACL DEFICIENT KNEE--MRI AND BIOMECHANICAL MODELING Principal Investigator & Institution: Buchanan, Thomas S.; Professor and Director; Mechanical Engineering; University of Delaware Newark, De 19716 Timing: Fiscal Year 2002; Project Start 01-JUN-1999; Project End 31-MAY-2004

Studies 9 Summary: The overall goal of this work is to provide a detailed understanding of the effect of anterior cruciate ligament injury on knee movement in those who compensate well for the injury and those who do not. Some persons (copers) are able to fully compensate for the absence of the anterior cruciate ligament (ACL) while others (non- copers) are not. Non-copers demonstrate quadriceps femoris weakness, and use kinematic, kinetic, and muscle activity patterns that stiffen the knee joint for stability. They accomplish the joint stiffening via general cocontraction of the muscles around the knee and by reducing the force with which the foot hits the ground. Copers have no quadriceps weakness, normal ground reaction forces, and possess an ability to coordinate the activity of the lower extremity muscles to efficiently distribute control of the knee among the hip, knee and ankle while maintaining normal knee motion. Even using sophisticated motion analysis techniques, copers are indistinguishable from uninjured subjects. A new approach to in vivo analysis of musculoskeletal dynamics uses Cine-phase contrast (Cine-PC) magnetic resonance imaging (MRI) to image and track the moving knee. Cine-PC MRI, a non-invasive technique, is capable of measuring 3D muscle fiber and skeletal velocity, in vivo, during dynamic tasks. Through integration, 3D musculoskeletal movement can be tracked. A combination of the use of this new technology and conventional MRI, electromyography, and musculoskeletal modeling will provide a unique opportunity to elucidate the compensation strategies employed by the copers. There are two aims to this proposal. Aim I is to identify differences in knee kinematics, ligament lengths, tendon lengths, and muscle activation patterns of ACL deficient patients using Cine-phase contrast MRI and electromyographic analysis that characterize the mechanisms with which the copers, in altering their muscle activation pattern, alter their knee joint kinematics in order to stabilize their knees. Aim II is to identify differences in muscle activation patterns in ACL deficient copers and non-copers using electromyography and biomechanical modeling. Patient specific models of the ACL deficient knee using T1-weighted MRI will be developed and used as input to a biomechanical analysis. Previous work suggests that patients with ACL deficiencies balance knee joint loads between muscles and ligaments using a strategy that is different than that employed by unimpaired subjects. This will be examined for copers and non-copers in this study. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: ALTERATIONS IN NEUROMUSCULAR FUNCTION FOLLOWING BURNS Principal Investigator & Institution: Martyn, J A.; Professor; Massachusetts General Hospital 55 Fruit St Boston, Ma 02114 Timing: Fiscal Year 2003; Project Start 01-DEC-1983; Project End 31-MAR-2007 Summary: (provided by applicant): Muscle weakness accompanies all forms of critical illnesses including burns, resulting in hypoventilation, dependence on respirators, and decreased mobilization, all of which lead to increased morbidity and mortality. The loss of muscle strength is out of proportion to loss of muscle mass. It is hypothesized (a) since prolonged open channel time of acetylcholine receptors (AChRs) due to congenital mutations of AChRs results in muscle weakness, the weakness of muscles in close proximity to bums is related to the expression of gamma subunits containing an \"immature\" isoform of AChRs, which also have a longer mean open-channel time; (b) that akin to that seen in many congenital muscular dystrophies (CMDs), the weakness in muscles at sites distant from burn occurs as a result of changes in muscle membrane structural components termed dystrophin associated complexes (DACs). It is postulated that the pathophysiological bases for the neuromuscular changes following burns are

10 Muscles related to (a) decreased signaling via agrin, important for clustering, expression, and maturation of the AChRs, and (b) decreased growth factor signaling via Akt/PKB, important for stabilization and maintenance of DACs, respectively. Related to the above: Specific Aim 1 tests the hypotheses (1) that muscles in close proximity to burn injury express an immature isoform of AChRs at the neuromuscular junction (NMJ), resulting in aberrant neurotransmission, (2) that these AChR changes are related to increased expression of iNOS, resulting in decreased signaling of agrin, and (3) that iNOS inhibitors and/or exogenous agrin will reverse the AChR changes and enhance muscle function. Specific Aim 2 tests the hypothesis that the diminished contractility of skeletal muscle at sites distant from bum is due to changes in muscle membrane DAC, independent of AChRs, since AChRs are unaltered at sites distant from burn. Muscle membrane costamere integrity will be determined by confocal microscope. Biochemical fractionation techniques (velocity gradients) will be utilized to detect molecular localization of each DAC component (dystrophin, dystroglycan, caveolin-3, and integrin), since abnormal localization of these membranes will result in disruption of costamere integrity. Specific Aim 3 using the rat in vivo model and the in vitro cell culture model, tests the hypothesis that bum injury-induced malformation of DAC is due to decreased pro-anabolic signaling via Akt/PKB. Specific Aim 4 tests the hypothesis that the attenuated Akt/PKB activity at sites distant from burn can be rectified by parenteral IGF-I or adenovirus transfer of Akt/PKB, both of which will restore the DAC integrity and function to normal. Delineation of the pathophysiology of burn-induced muscle dysfunction will provide a scientific rationale for therapeutic approaches to prevent muscular complications of burns. These mechanistic studies will also help to understand the molecular etiology of other acquired and congenital diseases of muscle, which affect muscle function in a vast number of adult and pediatric patients. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: ANATOMICAL SPECIALIZATIONS OF THE HUMAN PHARYNX Principal Investigator & Institution: Mu, Liancai; Otolaryngology; Mount Sinai School of Medicine of Nyu of New York University New York, Ny 10029 Timing: Fiscal Year 2002; Project Start 01-FEB-2001; Project End 31-JAN-2005 Summary: (Applicant's abstract): What variables in the neuromuscular properties of the human pharynx make some patients more susceptible to aspiration, obstructive sleep apnea (OSA), acid reflux, cricopharyngeal spasm and other disorders of the pharyngeal region? In most mammals (and neonatal humans) the respiratory system is protected by overlapping the epiglottis and soft palate, however with separation of these structures the human at risk of aspiration, and this is often the cause of death in the elderly and neurologically impaired. At present the basic neuromuscular specializations of the human pharynx are poorly understood. In preliminary work numerous novel observations were made, one example is that of the human cricopharyngeus (CP) muscle; That the CP receives its innervation from multiple nerves, each of which supplies a distinct region within the muscle, and that it contains specialized muscle fibers. One of these, slow tonic muscle fibers (STMF) has a unique physiology. STMF are extremely rare in mammals but preliminary work has shown that they are widespread in human upper airway structures including the tongue and larynx. Moreover the particular distribution of these fiber suggests that they are directly related to distinct biomechanics. The proposed work will focus on clarifying the peripheral organization patterns of the human pharyngeal plexus and characterizing the intrinsic properties of the CP and the muscles surrounding the pharynx to answer the questions: what

Studies 11 anatomic specializations are present that appear specific to humans and possibly speech and swallowing related? What variations in these specializations correlate with certain ethnic (black males OSA), genders (males reflux, OSA) and especially geriatric (CP spasm and aspiration) susceptibility to specific disorders? All studies will be done in human post-mortem tissue. The motor and sensory nerve supply to the pharyngeal region will be studied using Sihler's stain. An additional hypothesis to be tested is that the human glossopharyngeal nerve (IX) provides motor innervation not only to the traditionally described stylopharyngeus, but also to the CP and pharyngeal constrictor muscles as demonstrated by our preliminary studies. This will be studied by triple approaches: Sihler's stain whole-mount acetylcholinesterase (AChE) and silver stain, and Karnovsky-Roots' method. Another hypothesis to be tested is that most swallowing- related muscles are specialized and composed of neuromuscular compartments (NMC) as functional requirements. Our preliminary studies provided evidence for the existence of the NMC within the human CP inferior constrictor and geniohyoid muscles. In addition, the human CP appears to be a specialized skeletal muscle as it contains slow tonic and a-cardiac myosin heavy chain isoforms that are not normally present in limb muscles. The muscular specializations of the upper esophageal sphincter, pharyngeal constrictor and suprahyoid muscles will be explored using a wide variety of histochemical, immunohistochemical, electrophoretic and immunoblotting techniques The muscle fiber architecture, distribution of the fiber types and the major and unusual myosin heavy chain isoforms will be studied. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: ANKLE STRENGTHENING TO IMPROVE GAIT AND FUNCTION IN CP Principal Investigator & Institution: Engsberg, Jack R.; Associate Professor; Neurological Surgery; Washington University Lindell and Skinker Blvd St. Louis, Mo 63130 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-MAY-2005 Summary: (provided by the applicant): Many treatments exist to improve the gait and function of persons with Cerebral Palsy (CP). Despite recognizing that muscle weakness is a major impairment in CP, none of the treatments have the direct aim of strengthening muscles. Our results from an NIH investigation (R01-NS035830) indicated high correlations between ankle strength and function, with greater strength correlated with higher function. The idea of strengthening muscles has been controversial for safety issues. The purposes of this pilot investigation are to: 1) establish sample sizes for a future randomized clinical trial determining if intensive ankle strength training programs can improve strength, gait, and function without increasing spasticity; and 2) investigate potential safety issues arising from the training programs. Aim 1: Establish sample sizes for a future randomized clinical trial determining if intensive ankle strength training programs can improve ankle strength, gait, and function without increasing spasticity. Twenty ambulatory subjects with spastic diplegia CP will be randomly assigned to one of 4 groups: 1) Dorsi-flexor strength training group; 2) Plantar-flexor strength training group; 3) Dorsi-plantarflexor strength training group; and 4) the group undergoing no intensive strength training program. Subjects in the strength training groups will participate in a 12-week progressive, resistance strength- training program. Prior to, and at the end of the training program, all subjects will be objectively assessed for ankle Plantar-flexor spasticity, ankle strength, gait, and GMFM. The data will be used in a repeated measures power analysis to establish sample sizes for the clinical trial. Aim 2: Investigate potential safety issues arising from the training programs. The aim has 2 components. The first is the weekly measurement of Plantar-

12 Muscles flexor spasticity and tightness. The data will permit continuous monitoring of spasticity and tightness, and alert investigators to potential problems during each subject's participation. In the second component, both the pre- and post-intervention measures, and the weekly monitoring of spasticity and tightness, will determine if potential changes could be a concern for the future clinical trial. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: ANTERIOR CRUCIATE LIGAMENT-FUNCTIONAL BIOMECHANICS Principal Investigator & Institution: Andriacchi, Thomas P.; Professor; Mechanical Engineering; Stanford University Stanford, Ca 94305 Timing: Fiscal Year 2002; Project Start 01-APR-1988; Project End 31-MAY-2005 Summary: The long-term goal of this project is to provide information that can be applied to the prevention and treatment of injury to the anterior cruciate ligament (ACL) of the knee. The annual incidence of acute ACL disruptions is approximately 1 in 3000. Treatment of the ruptured anterior cruciate ligament is often complicated by the difficulty in predicting from passive physical examination of the knee which patients will be functionally impaired by the loss of this ligament and which patients will have minimal symptoms. Is it possible that altered patterns of locomotion dynamically compensate for loss of the ACT? Quantifying the relationship between altered patterns of locomotion and changes in the anterior-posterior displacement (AP) and internal- external rotation (IE) of the knee is a fundamental step towards answering this question. This information is clinically important since the AP and IE components of knee motion influence strains in secondary restraints (to anterior laxity) such as the medial meniscus of altered patterns of locomotion for ACL deficient knees. A newly developed point cluster technique will be used to quantify knee kinematics during locomotion. The first hypothesis will test if altered patterns of locomotion (characterized by the magnitude of the moment generated by net quadriceps/knee flexor muscles) are correlated with AP and IE displacements at the knee. Another consideration in this study is the possibility that individual anatomical variations can influence the effect of altered patterns of locomotion on knee kinematics. Previous work has implicated the extensor mechanism as a possible cause of these adaptations. The second hypothesis will test if the magnitude of the altered pattern of locomotion (defined by the reduction from normal) in the net quadriceps/knee flexor moment) is correlated with knee extensor anatomy. This study will generate fundamental new information on the patient's ability to dynamically control anterior posterior stability of the knee joint in the absence of anterior cruciate ligament. This study will also help to identify critical variables that should be considered in a larger prospective clinical outcome study. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: BINOCULAR COORDINATION IN MONKEYS WITH STRABISMUS Principal Investigator & Institution: Das, Vallabh E.; Neurology; Emory University 1784 North Decatur Road Atlanta, Ga 30322 Timing: Fiscal Year 2004; Project Start 01-FEB-2004; Project End 31-JAN-2007 Summary: (provided by applicant): Binocular alignment must be maintained in the horizontal, vertical and torsional planes to ensure binocular sensory fusion. Normal development ensures binocular alignment during fixation and binocular coordination during eye movements. Unfortunately, abnormal visual experience during development usually leads to ocular misalignment (strabismus). In fact, various studies have reported

Studies 13 the incidence of strabismus to be about 2-5% of the infant population. Data from strabismic humans and from strabismic monkeys in our laboratory have shown that ocular misalignment is accompanied by a lack of conjugate eye movements. Though strabismus is most often associated with a horizontal misalignment, often a combined horizontal, vertical and torsional misalignment is observed. Along with the static horizontal, vertical and torsional misalignment, there appears to be substantial dynamic cross-talk between the principal eye movement planes. In the clinical literature these apparent cross-axis interactions are usually described as 'A' and 'V' patterns of strabismus. Unfortunately, there is a lack of understanding of the neural or mechanical bases for these cross-axis movements, the putative relationship or lack thereof to the neural control of horizontal, vertical or torsional eye movements and the relationship to the etiology of the strabismus. Competing hypotheses include static malpositioning of extraocular muscle pulleys, sideslip of extraocular muscles and muscle pulleys, torsional control of eye movements gone awry leading to apparent muscle dysfunction and finally simply unexplained overaction/underaction of individual extraocular muscles. The goal of our studies is to clarify static and dynamic properties of cross-axis movements and examine its source in animals with a sensory induced strabismus. Our approach will include structural imaging of extraocular muscle to determine role of muscle pulleys; behavioral experiments to examine control of torsion and Listing's laws; neurophysiological experiments to examine the role of motor and pre-motor structures in the brain and biomechanical modeling of extraocular musculature to simulate experimental data. Completion of our studies will be of benefit to the understanding and treatment of certain types of strabismus. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: BINOCULAR MATCHING AND DISPARITY VERGENCE. Principal Investigator & Institution: Stevenson, Scott; Vision Sciences; University of Houston 4800 Calhoun Rd Houston, Tx 77004 Timing: Fiscal Year 2002; Project Start 10-APR-2001; Project End 31-MAR-2004 Summary: (adapted from applicant's abstract): Proper alignment of the eyes is essential for clear, single vision. Misalignment during early development can lead to amblyopia, a permanent visual impairment. Eye alignment during gaze changes is determined both by the anatomical organization of extraocular muscles and by the coordinated, visually- guided control of those muscles. The detection and correction of alignment errors from binocular comparison of retinal images is referred to as Disparity Vergence, and has both reflexive and voluntary aspects. The reflexive component of disparity vergence corrects horizontal, vertical and cyclotorsional errors of alignment, while voluntary control is restricted to horizontal vergence. This project is concerned with the visual information processing that provides the basis for reflexive disparity vergence, as revealed by vertical vergence responses. Previous work by the Principal Investigator has shown that the vertical vergence controller can extract vertical disparity signals from dynamic random dot stereograms, but that vertical vergence is not influenced by visual attention or subject effort and often occurs without conscious awareness. These movements thus reflect visual processes that are binocular, most probably cortical, but pre-conscious. The experiments in this project provide a way to study processing at an intermediate stage of the visual system. These processes are central to the control of eye alignment, but cannot be studied with conventional psychophysical techniques because they do not necessarily contribute to visual perception. An eye tracking device is used to detect small changes in eye alignment made in response to imposed vertical image misalignment, allowing for measurement of the vergence system's sensitivity to a

14 Muscles variety of image parameters. Proposed experiments will determine the role of contrast, spatial and temporal frequency, and visual feature type in the control of reflexive vergence eye movements. Measurements in subjects with abnormal binocularity will follow up on preliminary evidence that reflex vergence is intact in some cases of stereoblindness. Comparison to results from conventional psychophysical sensitivity measures will highlight differences between early (pre-conscious) and later (perceptual) visual processes. The long-term benefit of this research will be improvements in the diagnosis and treatment of binocular visual disorders of eye alignment and depth perception. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: BIOARTIFICIAL MUSCLES FOR GENE THERAPY Principal Investigator & Institution: Vandenburgh, Herman H.; Associate Professor; Cell Based Delivery, Inc. 4 Richmond Square, 5Th Fl Providence, Ri 02906 Timing: Fiscal Year 2002; Project Start 15-APR-1998; Project End 31-MAR-2003 Summary: Recombinant proteins are used to treat a number of human disorders including diabetes, neutropenia, anemia, and musculoskeletal disorders of the elderly. Muscle atrophy and bone wasting associated with aging can be attenuated using growth hormone (GH), insulin-like growth factor-1 (IGF-1), and parathyroid hormone (PTH) but delivery by daily injection is problematic since the proteins degrade rapidly in vivo, are expensive to manufacture, and have detrimental side effects when delivered at pharmacological doses. Cell based delivery of proteins from genetically-modified implanted cells may provide a more effective and cost-saving alternative. The long-term objective of this project is to develop and optimize the surgical techniques for reversible delivery of proteins from bioartificial muscle platforms. Muscle stem cells (myoblasts) can be isolated by simple needle biopsy and genetically modified to express foreign proteins. When tissue engineered in vitro into skeletal muscle-like bioartificial muscles (BAMs) and implanted in vivo, they serve as a long-term delivery system for biologically active proteins. Advantages of this technology over currently used injected myoblasts or plasmid DNA gene therapy techniques include efficient in vitro fusion of myoblasts into BAMs, preimplantation monitoring of growth factor secretion levels, and reversibility. BAMs secreting recombinant human GH (rhGH) and engineered from a murine C2C12 muscle cell line successfully attenuate skeletal muscle disuse atrophy when implanted subcutaneously under tension in mice. In the current project, primary myoblasts from inbred Fisher 344 rats will be transduced to constitutively express rhGH or IGF-1 using retroviral expression constructs under the control of the LTR viral promoter. New replication defective retrovirus expression vectors with the GH, IGF-1, and PTH genes under control of the regulatable human skeletal alpha -actin (HSA) promoter will also be constructed. BAMs from transduced primary rat myoblasts will be engineered using previously developed protocols and their morphology and protein secretion rates evaluated in vitro and in vivo. Rat BAM (R-BAM) myofiber survival, differentiation, innervation, and vascularization in subcutaneous and muscular sites will be studied by quantitative histological, immunocytochemical and biochemical techniques. The ability of GH, IGF-1 and PTH secreted from R-BAMs to attenuate muscle atrophy and bone wasting will be assessed in hindlimb unloaded adult Fisher rats. New therapeutic treatments with recombinant proteins for chronic musculoskeletal wasting disorders will lead to enhanced quality of life and reduced costs in this 150 billion dollar annual U.S. healthcare market. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

Studies 15 • Project Title: BIOMECHANICALLY BASED SHOULDER REHABILITATION STRATEGIES Principal Investigator & Institution: Ludewig, Paula M.; Phys Med and Rehabilitation; University of Minnesota Twin Cities 200 Oak Street Se Minneapolis, Mn 554552070 Timing: Fiscal Year 2003; Project Start 01-JUN-2003; Project End 31-MAY-2008 Summary: (provided by applicant): The candidate's long-term career objective is to investigate biomechanical factors contributing to musculoskeletal dysfunction, in order to refine current clinical treatment approaches and develop novel scientifically founded rehabilitation interventions. A five-year research and training plan developed with the primary mentor and a team of experienced scientists will expand the candidate's scientific background in biomechanics and musculoskeletal modeling, enrich her direct research skills and experience in the scientific process, promote integration of research findings to clinical practice, and advance skills necessary for becoming an independent investigator. The long-term objective of the research plan is to develop and test the effectiveness of biomechanically based rehabilitation strategies for improving upper extremity function and reducing pain and disability in persons with shoulder pathologies related to abnormal shoulder movement patterns. In-vivo 3-D full shoulder complex kinematics (thorax, clavicle, scapula, and humerus) during arm elevation will be collected from healthy and symptomatic subjects and integrated with a state of the art shoulder model to describe the 3-D muscle function of selected shoulder muscles, allowing comparisons among muscles for their relative biomechanical ability to reduce shoulder kinematic deviations, or contribute to deviations if producing excess or inadequate force (Aim 1). Computerized Tomography scans of the shoulder complex structures will be taken and imaging data combined with the kinematic data, allowing determination of the effects of abnormal kinematics on the available volume of the subacromial space, providing insight into how specific kinematic deviations create impingement of soft tissue structures (Aim 2). Determining and demonstrating muscle activations that can improve scapular kinematics and reduce subacromial impingement can provide a template for scientifically based intervention approaches that can be further tested through clinical trials. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: BODY COMPOSITION CHANGES IN THE ELDERLY--SARCOPENIA Principal Investigator & Institution: Baumgartner, Richard N.; Professor; Medicine; University of New Mexico Albuquerque Controller's Office Albuquerque, Nm 87131 Timing: Fiscal Year 2002; Project Start 30-SEP-1993; Project End 31-MAY-2004 Summary: (Adapted from Investigator's Abstract) The main objectives of this competing continuation application are to develop methods of estimating the prevalence and incidence rates of sarcopenia, or deficient relative muscle mass, and to determine sex and ethnic differences in risk factors and consequences of sarcopenia in community- dwelling elderly. It is accepted that muscle mass and strength are gradually lost with age. Because there are few methods of quantifying muscle mass in population studies, and criteria for defining \"deficient\" muscle mass remain to be established, estimates of the prevalence and incidence of sarcopenia are lacking and the extent of the public health problem posed is unknown. Age-related loss of muscle mass is undoubtedly multifactorial. Although a variety of possible mechanisms and etiological factors have been indicated, there are few data for multivariate associations of risk factors with sarcopenia. Sarcopenia is believed to be a major factor associated with physical functional impairment, and a number of studies have reported that indicators of muscle

16 Muscles strength and function (e.g., balance, gait speed, etc.) are associated with disability and falls in elderly people. There are few reports, however, for direct associations between sarcopenia and disability or falls. Sex and ethnic differences in rates of loss for muscles mass and strength, risk factors and consequences remain to be established. The proposed study will establish methods for defining sarcopenia using cross-sectional data collected previously in the New Mexico Elder Health Survey (NMEHS, 1993-1996, n = 883) and reference data to be collected for a population-based sample of 300 young adults 20 to 40 years of age. Risk factors and long-term consequences of sarcopenia will be studied using 4 to 10 years of follow up data by continuation of the New Mexico Aging Process Study (NMAPS, current n = 404). The following variables have been measured in the NMAPS since 1993: muscle mass from dual-energy X-ray absorptiometry, serum nutrient and hormone levels (e.g. free-T, estrone, IGF1, DHEAs, leptin), dietary intake, physical activity and resting energy expenditure, cognitive and physical functional status, disability, incident falls and morbidity. Data collected for these variables will be extended another 5 years. The NMEHS included Hispanic elderly men and women: 200 new Hispanic participants will be recruited in the NMAPS to further facilitate ethnic comparisons for risk factors and consequences of sarcopenia. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: BONE GROWTH, PERIOSTEAL MIGRATION AND MUSCLE FUNCTION Principal Investigator & Institution: Herring, Susan W.; Professor & Acting Chair; Orthodontics; University of Washington Grant & Contract Services Seattle, Wa 98105 Timing: Fiscal Year 2003; Project Start 01-MAY-1990; Project End 31-MAR-2008 Summary: (provided by applicant): Our overall goal is to clarify the influence of function on the growth and ultimate morphology of the head. The mechanical environment influences skull growth at every level from individual cells to gross structure. Although usually neglected, soft tissues such as muscles, ligaments and cartilages play a critical role in cranial mechanics and growth. This proposal focuses on two ways in which soft tissue mechanics may direct the growth of skull bones, using the pig as a model. First, the osteogenic activity of the periosteum is linked to its blood supply, which originates from muscles and ligaments. We hypothesize that the deformation of these soft tissues during function can modify periosteal perfusion. In Specific Aim 1, new methodology will be employed to map the three-dimensional deformation of muscles and ligaments during awake mastication and to test whether buccinator contraction places significant pressure on the alveolar periosteum. Specific Aim 2 addresses the periosteal vascular system directly with both immunocytochemical assays of cellular activity and in vivo measures of blood flow. These studies will provide evidence for or against a causal linkage between soft tissue behavior and periosteal perfusion. The second way in which soft tissues may direct skull growth involves the nasal septum. Despite being an unmineralized cartilage, the septum has been considered an important mechanical support of the face. Moreover, forces generated by septal growth are claimed to separate the sutures between facial bones, causing compensatory growth. These assertions have never been tested directly. In Specific Aim 3 a novel indwelling transducer will be used to reveal the mechanical loading pattern of the septum and to investigate the timing of its growth in relation to that of facial sutures. Specific Aim 4 will test the mechanical plausibility of the hypothesis that the septum controls facial growth by comparing the viscoelastic stiffness of the cartilaginous septum to the resistance of the facial sutures. Taken together, these studies will develop new techniques for monitoring soft tissue function, provide fundamental new information

Studies 17 about the mechanical behavior of the head, and test hypotheses about how soft tissues influence skull growth. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: CARDIAC CONTRACTILE PROTEIN COOPERATIVITY Principal Investigator & Institution: Tobacman, Larry S.; Professor; Internal Medicine; University of Iowa Iowa City, Ia 52242 Timing: Fiscal Year 2002; Project Start 01-AUG-1987; Project End 31-JUL-2006 Summary: (provided by applicant): This proposal concerns the mechanism controlling the repetitive contraction and relaxation of the heart, at the level of the protein assemblies that comprise the contractile apparatus. Cyclical variation in the calcium concentration causes calcium binding to and dissociation from troponin, which interacts with the other thin filament proteins tropomyosin and actin so that the thin filament switches between states that will or will not support contraction. A comparable system also exists in skeletal muscles. The long term goal of this work is to understand how troponin and tropomyosin exert this direct regulation, because of its fundamental physiological importance, because this regulation is altered in disease states, and because this is a potential target for therapy. Tropomyosin has an extended coiled-coil structure, and in striated muscles it spans seven actins. We will examine striated muscle alpha-tropomyosin segment by segment, to test hypotheses concerning its function and structure, to circumvent previous difficulties in understanding tropomyosin when using the entire molecule, and to evaluate evidence for major functional heterogeneity within this elongated protein. This will involve host-guest studies of function, and additionally both structural and functional studies of tropomyosin fragments. The structure and conformation of the thin filament also will be investigated by electron microscopy with 3-D reconstruction. This will involve filaments with altered forms of tropomyosin or troponin C (the calcium binding subunit), and short actin filaments created with the filament severing protein gelsolin. Solution studies of these altered filaments will be correlated with the structural results, and used to investigate the conformational transitions of the thin filament, and the spatial propagation of these transitions. Further, permeabilized muscle fibers containing altered forms of TnC will be used to investigate the mechanism of calcium-dependent cooperativity in the intact sarcomere. Finally, the mechanism of cooperative thin filament activation also will be investigated with statistical mechanical modeling of the effects of the non-homogeneous relationship between the regulatory proteins and the seven actins with each regulatory unit. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: CARDIOVASCULAR SEQUELLAE OF RESPIRATORY MUSCLE WORK Principal Investigator & Institution: Dempsey, Jerome A.; Professor; Population Health Sciences; University of Wisconsin Madison 750 University Ave Madison, Wi 53706 Timing: Fiscal Year 2002; Project Start 01-SEP-1977; Project End 31-MAR-2007 Summary: (provided by applicant): We propose to determine the physiological and clinical importance of metaboreflexes originating in the inspiratory and expiratory respiratory muscles in regulating blood flow and its distribution at rest and exercise. The rationale for this proposal is based on findings in humans which show that: a) mechanical unloading of the respiratory muscles in heavy exercise causes a reduction in stroke volume and cardiac output and vasodilation and increased blood flow in locomotor muscles; b) that fatiguing the diaphragm causes a time-dependent increases

18 Muscles in muscle sympathetic nerve activity (MSNA) in the resulting limb; and c) that central inspiratory motor output has no influence on MSNA in the intact human. Aim 1: We will voluntarily increase inspiratory and expiratory muscle effort in healthy humans at rest, during plantar flexion exercise and in hypoxia to determine the threshold and sensitivity of the respiratory muscle metaboreflex in response to progressive increases in respiratory muscle force output and fatigue. We will also determine the combined effects - additive or multiplicative - of combinations of forearm and diaphragm submaximal and fatiguing exercise. Outcome measures include: a) MSNA (via peroneal nerve microneurography); b) femoral arterial blood flow and vascular conductance (measured beat-by-beat with a Doppler ultrasound imaging technique). Aim 2: We will use local infusions of metabolites into the diaphragm and abdominal expiratory muscles in a chronically instrumented dog model in order to quantify the sensitivity and compensatory capabilities of the respiratory muscle metaboreflex and its effect on blood flow distribution at rest and exercise. This animal model will also be used to address the effects of the limb locomotor muscle metaboreflex on distribution of blood flow to the respiratory muscles during exercise. Aim 3: In patients with chronic heart failure of varying etiology, we will apply ventilatory assist in the form of pressure support or proportional assist mechanical ventilation to determine the influence of respiratory muscle work and intra-thoracic pressure on exercise performance, on stroke volume and cardiac output and on limb locomotor muscle blood flow and vascular resistance at rest and exercise. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: CATCH: MECHANOCHEMISTRY AND REGULATION IN SMOOTH MUSCLE Principal Investigator & Institution: Siegman, Marion J.; Professor; Physiology; Thomas Jefferson University Office of Research Administration Philadelphia, Pa 191075587 Timing: Fiscal Year 2002; Project Start 01-DEC-1994; Project End 31-JUL-2004 Summary: A unique and important characteristic of all smooth muscles (mammalian and invertebrate) is the ability to vary the energy cost of force production and maintenance during the time course of isometric contractions. The basic mechanisms controlling force maintenance, or tone in smooth muscle are not well understood. Our discovery that the phosphorylation state of twitchin (a mini-titin) regulates catch and force production in the anterior byssus retractor muscle (ABRM) of Mytilus edulis, has opened the way to new studies described here on the molecular basis of its function. Our overall goal is to determine the mechanisms whereby twitchin controls actin- myosin interaction and resulting mechanical output. The ubiquitous presence of twitchin in invertebrate striated, smooth catch and phasic muscles suggests that twitchin may serve as a regulator in all of these muscle types. The understanding of the basic mechanisms underlying this regulation may very well provide new insights on the control of other muscle types, especially mammalian smooth muscle, which shows very similar mechanical characteristics as the smooth muscles from invertebrates. In the proposed studies the smooth ABRM of M. edulis, and striated adductor of the sea scallop (P. magellanicus) will serve as experimental models. Intact and permeabilized invertebrate muscles will be used, as needed. The Specific Aims are to (1) Determine the mechanism by which twitchin phosphorylation gives rise to an increase in the detachment rate constant of the myosin crossbridge; (2) Determine the relationship between the degree of phosphorylation of twitchin and its mechanical effect in intact and permeabilized ABRM; (3) Test the hypothesis that the intrinsic rate of actin movement by myosin is attenuated by the presence of twitchin, and that the

Studies 19 phosphorylation of twitchin restores the intrinsic fast rate, using in vitro motility studies of actin movement on native thick filaments from ABRM and the body wall of C. elegans. Nucleotide turnover on the thick filaments will be measured in order to learn how twitchin phosphorylation alters the ATPase activity when calcium concentration is varied. (4) Determine of the effect of twitchin phosphorylation on fast striated scallop adductor muscle in order to learn whether this is a generalized mechanism for the modulation of crossbridge kinetics in invertebrate muscle. (5) Biochemical studies on twitchin will characterize the twitchin molecule and determine the mechanism by which its phosphorylation by protein kinase A controls the interaction of contractile proteins. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: CENTRAL AUDITORY PATHWAY OF THE MIDDLE EAR REFLEX Principal Investigator & Institution: Lee, Daniel; Otolaryngology; Univ of Massachusetts Med Sch Worcester Office of Research Funding Worcester, Ma 01655 Timing: Fiscal Year 2003; Project Start 15-JUL-2003; Project End 31-MAY-2008 Summary: (provided by applicant): The goal of this project is to understand the anatomic and physiologic features of interneurons of the mammalian middle ear muscle (MEM) reflex pathway. The neurons of this reflex coordinate the activity of the MEMs to protect the inner ear from intense acoustic stimuli as well as reduce masking. This reflex arc is composed of primary auditory afferents originating in the cochlea, a single or series of interneurons originating in the cochlear nucleus and ultimately synapsing on MEM motoneurons, and efferent fibers of the facial and trigeminal nerves that terminate on the stapedius and tensor tympani muscles, respectively. Although features of primary auditory afferents and the motoneuron efferents have been well characterized, little is known about the reflex interneurons. Which subdivision of the cochlear nucleus contains the reflex interneurons? Is there a single or a series of interneurons from the cochlear nucleus to the facial and trigeminal nuclei? For Aim 1, we will perform focal lesioning studies of the cochlear nucleus using kainic acid, an excitatory neurotoxin. We will correlate focal lesioning of the cochlear nucleus with loss of the MEM electromyography (EMG) response, to determine which division of the cochlear nucleus is involved in the MEM reflex pathway. Since the anatomical cell types of the cochlear nucleus subdivisions are well known, these studies will narrow down the identity of the cochlear nucleus interneurons. For Aim 2, we will examine the cochlear nucleus interneurons by double-injection experiments. We will inject retrograde tracer into either the stapedius or tensor tympani muscles to label their respective motoneurons, and, at the same time, inject an anterograde tracer into the cochlear nucleus to label the interneurons of the MEM reflex. Injections of the cochlear nucleus will be guided by our lesion studies described in Aim 1. Labeled projections from the cochlear nucleus will be identified as interneurons of the MEM reflex if they terminate on labeled MEM motoneurons. Such terminations would reveal a direct connection between the cochlear nucleus and the MEM motoneurons. Overall, the proposed project will improve our understanding of the brainstem connections that comprise the MEM reflex pathway. These findings may provide a basis for refining and extending our interpretation of clinical tests of MEM reflex integrity and brainstem auditory processing in humans. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: CHARACTERIZATION OF SKELETAL MUSCLE BY MR ELASTOGRAPHY Principal Investigator & Institution: An, Kai-Nan; Professor & Chair; Mayo Clinic Coll of Medicine, Rochester 200 1St St Sw Rochester, Mn 55905

20 Muscles Timing: Fiscal Year 2003; Project Start 01-APR-1999; Project End 31-MAR-2008 Summary: (provided by applicant): The goal of this proposal is two-fold: (1) to further develop and validate a technology, magnetic resonance elastography (MRE), for quantitatively imaging mechanical properties and tension distribution in muscle and (2) to apply the technique for in vivo evaluation of patients with four common, and clinically significant muscle disorders (spasticity, disuse atrophy, myofascial pain and a metabolic myopathy). These studies will employ a magnetic resonance imaging sequence with synchronous motion-sensitizing gradients to map propagating shear waves in the muscle. The technique will assess the mechanical properties of the muscle and its tension distribution. Specifically, the study can be divided into three specific aims. Aim 1: Optimize MRE methods of acquisition and analysis for the assessment of muscle, including electromechanical drivers, data acquisition techniques, and methods for image analysis. Advanced techniques for very rapid MRE assessment of muscle will continue to be developed. Aim 2: Validate the MRE assessment of muscle properties and tension with phantom, ex-vivo muscle, and Finite Element Modeling (FEM) techniques. Finite Element Analysis will be performed by using both phantom and bovine muscles to better correlate MRE wave-length findings as function of muscle properties, tension and fiber architecture. Aim 3: Study In Vivo Normal and Abnormal Muscle. The MRE technique will be applied in vivo to provide elastographic images of abnormal muscle with known disorders. The patient groups chosen for study are each important in their own right, and furnish unique information across the spectrum of muscular disease and dysfunction. Groups to be studied include individuals with new onset of spasticity following an ischemic, hemispheric stroke, disuse atrophy as a result of immobilization, metabolic (hyperthyroid) myopathy and myofascial pain for trigger point identification. The overall hypothesis of this work is that will bring benefits to both basic research and clinical care. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: CI-2 MODULATION OF SPINAL PROCESSES: SUPRASPINAL EFFECTS Principal Investigator & Institution: Foreman, Robert D.; Professor & Chair; Physiology; University of Oklahoma Hlth Sciences Ctr Health Sciences Center Oklahoma City, Ok 73126 Timing: Fiscal Year 2002; Project Start 01-SEP-1996; Project End 31-MAY-2007 Summary: (provided by applicant): The purpose of this study is to examine how neurons of propriospinal pathway(s) originating in the C1-C2 region process information from amygdala, subcoeruleus/parabrachial (SC/PB) nuclei, and vagal afferent fibers to modulate sensory-motor integration in the spinal cord. We previously demonstrated that chemical stimulation of C1-C2 neurons modulated spontaneous and visceral-evoked activity in lumbosacral spinal neurons and EMG activity of thoracic paraspinal muscles. Our preliminary data further demonstrate that chemical stimulation of C1 C2 neurons can strongly influence the activity of T3-T4 respiratory- related interneurons. Especially critical to this application are our preliminary results indicating that excitotoxic blockade of C1-C2 neurons with ibotenic acid, attenuated amygdalar and SC/PB modulation of lumbosacral spinal cells. The same lesion reduced vagal effects on lumbosacral neurons and paraspinal muscles. Our results challenge the assumption that descending pathways from supraspinal regions modulate activity of thoracic and lumbosacral neurons through direct projections only. The present application addresses the hypothesis that C1-C2 neurons process information from amygdala, SC/PB and vagal afferents. In turn, C1-C2 neurons strongly influence activity

Studies 21 of spinal sensory neurons, thoracic respiration-related interneurons, and somatomotor reflexes. We also hypothesize that amygdala and vagal afferents transmit information to C1-C2 neurons via SC/PB nuclei. Specific aims are designed to answer the following questions: 1) Are discharge patterns and activities of C1-C2 neurons affected by stimulating specific supraspinal nuclei? 2) Do neurons in C1-C2 segments process information from specific supraspinal nuclei to change sensory and integrative/motor activity in the spinal cord? 3) Do neurons in C1-C2 segments process information from vagal afferents to change integrative/motor and sensory activity in the spinal cord? 4) Do SC/PB nuclei relay information from amygdala and vagal afferent fibers to the C1- C2 segments to change sensory and integrative/motor activity in the spinal cord? Neurophysiological studies to examine extracellular discharge patterns and studies using c-fos as a marker of neuronal activation will be conducted in anesthetized rats. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: DEFINING HETEROPLASMY AT THE SINGLE MITOCHONDRION LEVEL Principal Investigator & Institution: Arriaga, Edgar A.; Assistant Professor; Chemistry; University of Minnesota Twin Cities 200 Oak Street Se Minneapolis, Mn 554552070 Timing: Fiscal Year 2002; Project Start 15-APR-2002; Project End 31-MAR-2006 Summary: (provided by applicant): Our long-term goal is to understand how mitochondrial DNA (mtDNA) mutations appear, propagate, and are distributed among the thousands of mtDNA molecules in a cell, a condition called heteroplasmy. While these mutations are associated with age-related diseases and the aging process, predicting the degree of heteroplasmy at which disease symptoms or age-related phenotypes will appear is practically impossible because the dynamics of heteroplasmy are not well understood. The central hypothesis of this application is that individual mitochondria, containing anywhere from 2 to 10 mtDNA copies, can be heteroplasmic, a condition resulting from the segregation of mtDNA molecules upon mitochondrial replication, and likely modified by the dynamic exchange of genomic material among mitochondria within a given cell. If this hypothesis is correct, a heteroplasmic mitochondrion will have both mutated and wild-type mtDNA, all the peptides encoded by the mitochondrial genome, and a normal mitochondrial membrane potential. We propose to develop the first bioanalytical technologies capable of investigating heteroplasmy in individual mitochondria. We will continue to use and improve upon an instrument based on capillary electrophoresis with laser-induced fluorescence detection (CE-LI F) to determine the properties of individual mitochondria that can then be collected and subjected to PCR amplification of their DNA. In addition, peptide profiles from mitochondria containing mutated mtDNA that will be determined by in situ matrix-assisted laser-desorption time-of-flight mass spectrometry will provide a more comprehensive characterization of heteroplasmy. As testing models, we will use NS-1 cells lines, two cybrid cell lines harboring 7522 and 4977 deletions, and rectus femoris and soleous muscles from Fisher 344 Rats, aged 6, 24, and 28 months. The NS-1 model will mainly be used to develop technologies and methods. The cybrid models have a defined degree of heteroplasmy (> 50 percent) and host deletions that omit the expression of the genes ND5, ND4, ND3, ND4L, COIII, A6, A8, and tRNAL, tRNAS, tRNAH, tRNAR, and tRNAG. In addition to these genes the cybrid hosting the 7522 base pair deletion further omits expression of the cytb, ND6, COIl (2), tRNAR, and tRNAK genes. These cybrid models will be used to study the progression of heteroplasmy in cell lines. The two muscle models will be used to study the progression of heteroplasmy along red ragged fibers that have been identified by (COX-, SDH++)

22 Muscles phenotype. Our individual mitochondrial determinations will be the basis for monitoring (1) the progression of heteroplasmy after the formation and propagation of a cybrid clone, and (2) the degree of heteroplasmy along skeletal muscle fibers. The data resulting from the cybrid and muscle tissue models will be used to refine existing mathematical models that predict the clonal expansion of heteroplasmy. The determination of mtDNA mutations at the single mitochondrion level in the cybrid and muscle models will bring us closer to uncovering the intricacies of heteroplasmy and its implications in disease and aging. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: DEVELOPMENT OF INNERVATION TOPOGRAPHY IN MUSCLE Principal Investigator & Institution: Laskowski, Michael B.; Biological Sciences; University of Idaho Moscow, Id 838443020 Timing: Fiscal Year 2002; Project Start 01-JUL-1988; Project End 31-MAY-2005 Summary: (From the Applicant's Abstract): Over the last several years, we have shown that motor neuron pools map onto muscles with a rostrocaudal positional bias. Detailed studies from our lab revealed that this topographic map is detectable in embryonic muscles upon first contact between nerve and muscle, and is partially restored after denervation. We have developed an important model of synaptic competition during reinnervation, where we can predict with 95 percent accuracy the survivor between two competing nerve terminals. We have also developed an in vitro model to identify muscle membrane-bound labels that may be responsible for the positional bias. We have found selective growth of embryonic spinal cord neurites on membranes derived from embryonic rostral or caudal muscles or from transgenic muscle cell lines bearing a heritable memory for rostrocaudal position. We have recently focused our attention on the Eph A/ephrin A subfamily of tyrosine kinase receptors as a class of candidate molecules that regulate neuromuscular topography. We have found that all five members of the ephrin A subfamily are expressed in embryonic muscles, and that membrane expression of ephrin A ligands progressively diminishes during postnatal development. We have further found that overexpression of ephrin A5 or deletion of ephrin A5 and A2 degrades the topographic map. We propose to build on this series of observations in three ways. First, we will study the physiological basis for the altered topographic map by ephrins A using intracellular recording and uptake of activity dependent dyes into living nerve terminals. Second, we will extend our in vitro model for innervation topography using a wide array of neurite growth assays. In particular, we will examine growth on membranes of two particularly selective muscles, the gluteus and serratus anterior where 87 percent to 95 percent of the neurites making a choice grew selectively on membranes of similar axial position. We will also explore selective neurite growth within compartments of a single muscle. Third, we will use this in vitro model to search for molecular guidance cues other than ephrin A ligands that may cooperate in establishing the neuromuscular map. This will include the use of ephrin A5 fusion proteins to block endogenous ephrin A ligands. In addition, we will isolate membranes from mutant mice where ephrin A5 or A2/A5 genes have been deleted. In both cases we will search for residual selective growth by spinal motor neurites as a first step toward isolation of additional guidance molecules. Results of these studies will provide unique insight into how neurites in the peripheral nervous system recognize and synapse with their positionally matched partners. We will also learn whether positional labels in the neuromuscular system are part of a general strategy for encoding position in the nervous system. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

Studies 23 • Project Title: DIABETES ENDOCRINOLOGY RESEARCH CENTER Principal Investigator & Institution: Barrett, Eugene J.; Professor; Internal Medicine; University of Virginia Charlottesville Box 400195 Charlottesville, Va 22904 Timing: Fiscal Year 2003; Project Start 01-MAY-2003; Project End 31-JAN-2008 Summary: (provided by applicant) We propose to establish a DERC within the University of Virginia Diabetes Center. The research base of the proposed DERC includes 43 scientists with distinguished research accomplishments in the fields of diabetes, endocrinology, immunology, cell signaling and vascular disease. Their research is organized in 3 thematic areas autoimmunity, insulin secretion and action and vascular complications of diabetes. These are areas of current strength and future growth for diabetes at Virginia. The proposed DERC will be a magnet to attract faculty and research support in diabetes and its complications to the institution. The proposed DERC will center around 5 scientific core laboratories, and the Pilot and Feasibility and Enrichment Programs. Organizational structure and grants management by an Administrative Core will support these activities. The Cores include a Genetics Core that will give DERC members facilities for genotyping, linkage analysis and generating speed congenics; a Biomolecular Research Core with tools for DNA sequencing and peptide and oligonucleotide synthesis as well as unique capabilities for protein sequencing using mass spectroscopy; an Animal Characterization Core that will facilitate detailed metabolic, imaging and behavior assessment of nutritionally, genetically, pharmacologically or behaviorally treated rodents as well as facilities for immune assay and phospho and fluoro imaging; a Cell and Islet Isolation Core that will provide facilities to isolate and characterize islets, adipocytes, and isolated muscles, as well as routine access to tissue culture reagents; and a Integrated Data Management Core that will bring mathematical tools to analyze, integrate and archive data from individual laboratories and from other Cores to enhance research programs of center investigators. These capabilities in our scientific cores promise to drive a period of discovery in diabetes and endocrine research at the University of Virginia. Pilot and Feasibility and Enrichment Programs will introduce new investigators to diabetes research in an environment of research excellence. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: DIETARY RESTRICTION, MT DNA ABNORMALITIES AND AGING Principal Investigator & Institution: Aiken, Judd M.; Professor; Animal Hlth & Biomedical Scis; University of Wisconsin Madison 750 University Ave Madison, Wi 53706 Timing: Fiscal Year 2002; Project Start 10-JUL-1995; Project End 31-JAN-2006 Summary: (Verbatim from application) Aging is recognized as an intricate web of global, physiological attrition. Many of the physiologically significant age-related changes are exhibited in non-replicative tissues such as brain, heart and skeletal muscle that rely heavily on oxidative metabolism for energy. In skeletal muscle, we hypothesize that mitochondrial genetic and enzymatic abnormalities, possibly secondary to life-long oxidative damage, may ultimately disrupt cellular processes or trigger cell death. The ensuing skeletal muscle fiber dysfunction or loss may contribute to sarcopenia, the age- related loss of skeletal muscle mass and function. We are addressing, by the in situ analyses of skeletal muscle from aged rodents, the question of the biological impact of mitochondrial abnormalities. Our studies suggest a specific sequence of events linking mtDNA deletions to sarcopenia. Concomitant with decreased muscle mass and fiber number, we have observed increases in segmental mitochondrial abnormalities that

24 Muscles contain specific rntDNA deletion mutations as revealed by laser capture microdissection and whole mitochondrial genome amplification. Muscle fibers harboring mtDNA deletion mutations often display atrophy, splitting and oxidative damage demonstrating a cellular impact of these abnormalities. These correlations suggest a causal role for mtDNA deletion mutations in sarcopenia. The aims of the present proposal are four- fold: 1) characterize ETS abnormalities, fiber atrophy, fiber splitting and oxidative damage during the progression of sarcopenia in selected rat muscles; 2) ascertain the cellular impact of age-associated ETS abnormal segments by gene expression profiling of laser-capture microdissected muscle fibers 3) Assess the effect of early- and adult- onset caloric restriction on the progression of sarcopenia and the accumulation of mitochondrial abnormalities in selected muscles of F344BNF1 rats; 4) determine whether mitochondrial genomes harboring deletion mutations are causally related to age-associated ETS abnormalities and subsequent cellular impact. The outcome of this work will shed additional light on the biological significance of these mutations and the effects they have on the age-related changes in muscle physiology and structure. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: DIFFERENCES IN SWALLOW MECHANICS IN INFANTS Principal Investigator & Institution: German, Rebecca Z.; Professor; Biological Sciences; University of Cincinnati 2624 Clifton Ave Cincinnati, Oh 45221 Timing: Fiscal Year 2002; Project Start 01-JUL-1998; Project End 31-JUL-2006 Summary: (provided by applicant): Swallowing requires the coordination of a large number of muscles; this complexity arises partly from the need for airway protection. In the previous funding period, we added to the understanding of muscle activity and oropharyngeal kinematics in infant deglutition. However, the role of the majority of muscles during emptying of the valleculae and in the transport of the bolus past the laryngeal opening or the natural stimuli that initiate the emptying of the valleculae over maturation is not well understood. Our preliminary data suggest that two distinct pathways of bolus movement exist, either around the epiglottis/laryngeal opening (in the newborn) or over it (by the age of weaning). However the timing of the transition, from one path to the other and the associated changes in the kinematics or motor patterns, are unknown. The decerebrate pig is an excellent model for studying vallecular emptying because this phase of the swallow can be isolated experimentally. We propose to apply our existing techniques both to this model and to intact animals, in order to answer the following questions. What natural stimuli initiate vallecular emptying, and do they change during maturation? What is the pattern of muscle activity during vallecular emptying in terms of the order and amplitude of muscle activation? Does change in the consistency of the bolus alter the motor pattern during vallecular emptying, and does this change over developmental time? Does epiglottal movement result from: (i) direct muscle contraction; (ii) indirect movement of the rest of the larynx, (iii) the mechanical action of food on the epiglottis, or a combination of all three? Current studies of human dysphagia and rehabilitation rely heavily on several older studies of oral function in adult man and animal; these studies did not have the means to examine the ontogeny of vallecular function in detail. The proposed study of the maturation of motor patterns will provide an important baseline for treatment strategies aimed at human infant dysphagia. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

Studies 25 • Project Title: DISEASE AND CONTINUOUS MYOFIBER REMODELING IN EOM Principal Investigator & Institution: Mcloon, Linda K.; Associate Professor; Ophthalmology; University of Minnesota Twin Cities 200 Oak Street Se Minneapolis, Mn 554552070 Timing: Fiscal Year 2002; Project Start 01-JUN-2002; Project End 31-MAY-2005 Summary: (Applicant's Abstract) Extraocular muscles (EOM) are spared or preferentially involved in various skeletal muscle diseases. We propose a novel and controversial alternative mechanism that may form the basis for the differences between extraocular muscles and limb skeletal muscles. We have strong preliminary evidence to suggest that there is ongoing, continuous myofiber remodeling in the adult extraocular muscles. This would involve both myogenic and apoptotic components. These conclusions are based on 4 lines of evidence. In this proposal we are asking questions to confirm our original observations. We have shown that the EOM continue to express cells positive for myogenic regulatory factors, such as myoD. Activated satellite cells are always present in adult EOM. BrdU labeling experiments using both 2 week and 4 week continuous labeling protocols followed by various brdU-free periods, a protocol that labels dividing cells, demonstrated mature myofibers with brdU-positive nuclei within them. Our working hypothesis is that there are mechanisms present in adult EOM that allow continuous satellite cell activation and division, resulting in either continuous remodeling of existing myofibers by fusion of new myoblasts with existing adult myofibers or formation of entirely new myofibers by the fusion of myoblasts with each other. This proposal asks the following questions: What are the mechanisms of myofiber remodeling in adult EOM? What is the time course and extent of fiber remodeling? What role does apoptosis play in myofiber remodeling and by what mechanism? How do surgical and chemodenervation manipulations simulating strabismus treatments cause changes in myofiber remodeling that may be affecting long-term surgical outcomes? The ability of adult EOM to continuously remodel provides a wealth of testable hypotheses for some long-standing enigmas involving the EOM and their preferential sparing or involvement in various muscle diseases. Ultimately, we hope to use this information to develop new therapeutic strategies.for the treatment of strabismus and other ocular and non-ocular muscle diseases. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: DO LIGHT CHAIN EXTENSIONS ENHANCE MUSCLE POWER OUTPUT? Principal Investigator & Institution: Maughan, David W.; Research Professor; Molecular Physiol & Biophysics; University of Vermont & St Agric College 340 Waterman Building Burlington, Vt 05405 Timing: Fiscal Year 2002; Project Start 01-AUG-2001; Project End 31-JUL-2005 Summary: Oscillatory power production is a general feature of striated muscle. Enhanced oscillatory power output is correlated with the presence of extensions of the amino terminus of myosin light chains. Homologous extensions exist in myosin essential light chains of vertebrate myocardium and the regulatory light chain of Drosophila jump and flight muscles. The exact function of these protein extensions is unknown, but preliminary results suggest that they augment power during contraction. Our central hypothesis is that the light chain extensions make molecular contacts that help pre-position the motor subunit of myosin near its target zone on actin for optimal interaction and power generation. Light chain constructs will be created in mouse myocardium and flies to assess the extent to which power output is diminished by

26 Muscles removing or replacing residues thought to be involved in thin filament interaction. The following hypotheses will be tested: 1) intact ventricular strips lacking the essential light chain N-terminal extension produce lower oscillatory power output at submaximal calcium levels than strips with full length light chains, 2) the essential light chain extension exhibits its effect on power only at in vivo lattice spacing, 3) the light chain extension exerts its effect on power by specific, electrostatic interactions with the thin filament, and 4) comparable alterations of the N-terminal extension of the regulatory light chain in Drosophila flight and jump muscles produce structural and functional phenotypes comparable to those observed in mouse hearts. Interfilament spacing, lattice order, and indices of myosin head alignment will be measured in both intact and demembranated (skinned) preparations using low-angle X-ray diffraction, aided by electron microscopy. Isometric force, unloaded shortening velocity, and dynamic stiffness (oscillatory power output) will be measured in skinned preparations under conditions in which the otherwise swollen lattice is restored by osmotic compression to its in vivo spacing. This research will contribute to understanding and treating human muscle diseases in which power output is compromised. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: DOES SPINAL MANIPULATION SPEED DETERMINE NEURAL RESPONSE Principal Investigator & Institution: Pickar, Joel G.; Research and Development; Palmer Chiropractic Universtiy 1000 Brady St Davenport, Ia 52803 Timing: Fiscal Year 2002; Project Start 01-AUG-2001; Project End 31-JUL-2004 Summary: The goal of this R21 PROJECT is two-fold: 1) to increase our understanding regarding the effects of spinal manipulation on the nervous system and 2) to seek a scientific basis for the continued investigation of the role of proprioceptors in the effects of spinal manipulation. The specific aim of this project is to determine if the speed of a spinal manipulation is an important determinant of the neural response from paraspinal muscle proprioceptors. Strong evidence supports using spinal manipulation to help patients with acute low back pain and neck pain. A theory common to the practice of spinal manipulation proposes that spinal manipulation alters paraspinal sensory input (ie, neural input from tissues of the vertebral column). Preliminary data demonstrate that spinal manipulative impulses stimulate proprioceptive afferents from lumbar paraspinal muscles. These afferents could contribute to the therapeutic effects of manipulation. The proposed experiments will determine how muscle spindles and Golgi tendon organs in lumbar paraspinal muscle respond to the time-varying impulse of a spinal manipulation. Slow and fast impulses will be given. A Fourier transform will be used to analyze, in the time domain, the force-time and displacement-time profile of each manipulative impulse. The resulting power spectra for a range of impulse durations will enable us to determine if the speed of a spinal manipulation is an important determinant of proprioceptive sensory input from paraspinal muscles during spinal manipulation. The experiments will provide information regarding the neural systems impacted by spinal manipulation and can lead to improved training methods for the proper application of spinal manipulation. Teaching the manual skill of spinal manipulation could be approached from the perspective of quantifying the velocity with which the clinician applies a spinal manipulation. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

Studies 27 • Project Title: DYNAMICS OF MUSCLE FORCE PRODUCTION DURING LOCOMOTION Principal Investigator & Institution: Roberts, Thomas J.; Zoology; Oregon State University Corvallis, or 973391086 Timing: Fiscal Year 2002; Project Start 01-APR-2000; Project End 31-MAR-2004 Summary: (adapted from Investigator's abstract) The mechanical and metabolic energetics of locomotion are ultimately determined by the mechanical properties of skeletal muscles and the pattern of contraction they undergo. The link between muscle properties and movement energetics is poorly developed because we lack information about how muscles contract in vivo. The proposed research will use a particularly suitable locomotor model to measure force, power and activity of muscles and tendons directly during running, walking and acceleration. Direct measurements of muscle contraction in vivo, measurements of tissue properties, and inverse dynamics will be used to determine how muscle contractile power is translated into movement. Independent measurements of muscle and tendon work will be used to test the hypothesis that tendon energy recovery supplies the majority of the positive work of movement during steady-speed level walking and running. The force-velocity and length-tension properties of muscles will be used to test the hypothesis that muscles operate at lengths and shortening velocities that allow for economical force production during steady-speed walking and running. The role of passive force development during movement will also be investigated to test the hypotheses that muscles produce force passively over a range of muscle lengths and velocities where active force capacity is low. These studies will provide insight into how the cellular and molecular properties of muscles and tendons determine the energetics and mechanics of normal gait. A basic understanding of muscle mechanical function during normal gait is important for developing rehabilitative therapies for individuals with musculoskeletal injuries or gait disorders, the design of prosthetic devices, and an understanding of the mechanical forces that influence the regulation of muscle properties. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: ESTROGEN EFFECTS ON CARDIOVASCULAR RESPONSE TO EXERCISE Principal Investigator & Institution: Kaufman, Marc P.; Professor of Internal Medicine & Human p; Internal Medicine; University of California Davis Sponsored Programs, 118 Everson Hall Davis, Ca 95616 Timing: Fiscal Year 2002; Project Start 01-FEB-2001; Project End 31-JAN-2006 Summary: (Applicant's abstract): Static and moderate dynamic exercise are known to increase heart rate, myocardial contractility, arterial blood pressure, breathing and muscle sympathetic nerve discharge. These effects, which are believed to increase the delivery of oxygen to metabolically active tissues (i.e., the exercising muscles), appear to be less in women than in men. This difference is often attributed to the effect of estrogen on neuronal function. Consequently, the aim of the experiments proposed in this application is to identify the effect of estrogen on \"central command\" and the muscle reflex, the two neural mechanisms responsible for evoking the autonomic responses to exercise. The proposed studies will be done in decerebrate unanesthetized female and male cats, which have been either ovariectomized or castrated, respectively two to four weeks prior to the experiment. In this preparation, the two neural mechanisms, central command and the muscle reflex, can be investigated separately without the influence of anesthesia. The effect of estrogen (i.e., 17-beta-estradiol) on the central command to

28 Muscles exercise will be studied while the cats are paralyzed with vecuronium, and will be evoked by both electrical and chemical stimulation of the hypothalamic and mesencephalic locomotor regions. Motoneuron discharge to agonist and antagonist hindlimb muscles will be recorded. The criterion for elicitation of central command will be \"fictive locomotion.\" Likewise, the effect of estrogen on the muscle reflex will be studied, but the cats will not be paralyzed. The muscle reflex will be evoked both while the hindlimb muscles are freely perfused and while they are ischemic. Dose response relationships for the effect of estrogen on both the cardiovascular and respiratory responses to central command and the muscle reflex will be determined. Moreover, studies will be extended to estrogen pretreatment with timed release pellets implanted into castrated male cats and ovariectomized females. In addition, the effect of microinjections of 17beta-estradiol into the hypothalamic and mesencephalic locomotor regions will be determined because preliminary data suggest that central command, but not the muscle reflex, is responsible for the estrogen-induced attenuation of the cardiovascular and ventilatory responses to exercise. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: EXPIRATORY MUSCLE ACTIVATION TO PRODUCE COUGH Principal Investigator & Institution: Dimarco, Anthony F.; Physiology and Biophysics; Case Western Reserve University 10900 Euclid Ave Cleveland, Oh 44106 Timing: Fiscal Year 2002; Project Start 01-FEB-1986; Project End 31-JUL-2004 Summary: (Applicant's abstract):Patients with spinal cord injury frequently suffer from respiratory complications due to their inability to cough and clear secretions. In recent animal studies, we have demonstrated that lower thoracic spinal cord stimulation (SCS) and magnetic stimulation (MS) results in the generation of large increases in airway pressure and high peak flow rates. These techniques, therefore, have the potential to produce an effective cough mechanism in spinal cord injured patients. The purpose of these studies is to resolve important basic science issues concerning these techniques in animal studies. In OBJECTIVE I, the efficacy of cough by these techniques will be assessed by radiolabeled clearance studies. In OBJECTIVE II, the pathway(s) by which the motor nerves innervating the expiratory muscles are activated during SCS and MS will be determined. The importance of motor root activation via stimulation of spinal cord pathways will be assessed by monitoring pressure generation before and after sequential section of the ventral roots. The specific pathways responsible for pressure generation will be localized anatomically by evaluating the effects of spinal cord section. Nerve compound action potentials will also be recorded from the motor roots during stimulation. In OBJECTIVE III, the electric field generated around and within the spinal cord during SCS and MS will be measured and used in conjunction with finite element analysis modeling techniques to determine optimum electrode and coil design. In OBJECTIVE IV, we will characterize the changes in expiratory muscle structure and function following upper motoneuron denervation. An effective cough is dependent upon optimal function of the expiratory muscles which are most likely atrophied in patients with spinal cord injury. Therefore, we will also assess the capacity or SCS and MS to maintain expiratory muscle function in a chronic animal model of spinal cord injury. In OBJECTIVE V, the safety profile of SCS will be assessed in chronic animals. The results of these studies should provide important information relevant to the potential use of these techniques in human clinical trials. Restoration of affective cough mechanism may allow patients with spinal cord injury to clear secretions more easily, reduce the incidence of respiratory complication and, ultimately, improve their life quality.

Studies 29 Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: EYELID SENSORIMOTOR NETWORKS Principal Investigator & Institution: Ledoux, Mark S.; Associate Professor; Neurology; University of Tennessee Health Sci Ctr Memphis, Tn 38163 Timing: Fiscal Year 2002; Project Start 04-AUG-2000; Project End 31-MAY-2004 Summary: Normal eyelid motor function depends on neurons that innervate the orbicularis oculi muscles that clone the eyes during blinks and levator palpebrae muscles that open the eyes. Neural structures afferent to orbicularis oculi and levator palpebrae motoneurons control the parameters of voluntary eyelid opening and closure, spontaneous and reflexive blinks, and eyelid activity that accompanies eye movements. The motor circuitry mediating spontaneous and reflex blinking is critical for the maintenance of normal ocular function and prevention of ocular injury. Disorders of the nervous system associated with abnormal blinking such as blepharospasm and apraxia of eyelid opening can produce significant functional disability including blindness. Lid retraction and decreased blink frequency seen in neurodegenerative disorders such as progressive supranuclear palsy can cause dry eye and exposure keratitis. Blepharospasm is an involuntary, typically bilateral, closure of the eyes secondary to spasmodic contractions of the orbicularis oculi musculature. Blepharospasm, although usually idiopathic, has been associated with structural lesions of the central nervous system, particularly the rostral brainstem and mesencephalon. Light sensitivity (photophobia) is a symptom with most patients. Some patients with blepharospasm have a history of irritative ocular stimuli such as blepharitis or dry eye; one hypothesis is that maladaptive responses to these stimuli are critical to the development of blepharospasm. Pharmacological, physiological, and postmortem-pathological evidence suggest that monoaminergic systems, particularly serotonergic, may play a role in the pathophysiology of blepharospasm. The neural circuits premotor to orbicularis oculi and levator palpebrae motoneurons will be defined anatomically in both rats and primates using both standard and viral transneuronal tracers. These experiments will also determine the relationship of orbicularis oculi premotor neurons to the central terminations of trigeminal afferents from the eyelid and cornea. The simultaneous use of two transneuronal tracers will localize neural structures critical to the bilateral coordination of orbicularis oculi and levator palpebrae motoneuron activity. Finally, the components of the orbicularis oculi premotor network activated either acutely or chronically by irritative ocular stimuli will be determined in rats. The data generated from these experiments will contribute to the development of models of eyelid motor function and dysfunction, improve understanding of clinical blink reflex testing and conditioning studies of the blink reflex, and provide important information regarding the cell-specific transport of viruses into rodent and primate nervous systems. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: FACTORS THAT MODIFY INSULIN ACTION Principal Investigator & Institution: Buse, Maria G.; Professor; Medicine; Medical University of South Carolina 171 Ashley Ave Charleston, Sc 29425 Timing: Fiscal Year 2002; Project Start 01-MAY-1978; Project End 31-MAR-2003 Summary: \"Glucose toxicity\" accounts for insulin resistance in uncontrolled Type I diabetes (IDDM) and contributes to insulin resistance in Type II diabetes (NIDDM). Sustained hyperglycemia or hyperinsulinemia cause insulin resistance; glucose and insulin act synergistically in down- regulating insulin-stimulated glucose transport. A

30 Muscles hypothesis to be tested in 3T3-Ll adipocytes is that glucose/insulin induced glucose transport desensitization reflects altered subcellular trafficking of the glucose transporter, GLUT4, which may involve impaired GLUT4 translocation and inappropriate association of GLUT4 containing vesicles (GCV) with the plasma membrane. Products of the hexosamine synthesis pathway (HNSP) have been implicated in glucose-induced insulin resistance; glutamine-fructose-6-P amidotransferase (GFAT) is the rate limiting enzyme and UDP-N-acetyl glucosamine (UDP-GlcNAc) the major product. The role of HNSP will be tested by examining whether conditions which increase or decrease flux via HNSP augment or mitigate, respectively, glucose induced insulin resistance. O-GlcNAcylation is a reversible process, involving O-glycosylation of proteins on Ser/Thr residues with monosaccharide GlcNAc. It usually involves phosphorylation sites and may be regulatory. Based on preliminary data in muscles of a mouse model of insulin resistance, over-expressing GLUTI in muscle, the hypothesis will be tested that increased flux via HNSP promotes O-GlcNAcylation of critical proteins involved in insulin- stimulated glucose transport. These may include GSV-associated proteins, possibly GLUT4 itself and/or proteins associated with GSV docking and fusion. Since adaptive regulation usually involves multiple sites, we will test the hypothesis that glucose-induced insulin resistance represents in part down-regulation of the insulin receptor (IR) signaling cascade, attempt to identify the major regulatory sites and critically assess the possible contribution of HNSP to the glucose effect. If warranted, the involvement of modulators of IR signal transduction, I.E. protein kinase C (PKC) isoforms, and candidate protein tyrosine phosphatases (PTP-ases: PTP-1B, SH-PTP2 and LAR) will be examined. GFAT activity is allosterically regulated by UDP-GlcNAc, and is modulated in vivo in muscle by the hormonal and metabolic milieu. The pre- and post- translational regulation of GFAT expression will be studied in muscles of rodent models. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: FAILED RESCUE OF OLD SKELETAL MUSCLE FROM ATROPHY Principal Investigator & Institution: Booth, Frank W.; Professor; Veterinary Biomedical Sciences; University of Missouri Columbia 310 Jesse Hall Columbia, Mo 65211 Timing: Fiscal Year 2002; Project Start 15-AUG-2001; Project End 31-JUL-2005 Summary: A common clinical problem is that in many nursing homes there are mobile and functioning aged individuals who, upon being subjected to one or more periods of immobility due to illness or injury, are unable to return back to mobility. Even with extensive rehabilitative therapy, many of these individuals are unable to recover to preinjury functioning levels. An animal model mimics this human condition. Both young and old rats who undergo a 10-day period of hindlimb immobilization exhibit disuse atrophy, but only skeletal muscle from young rats successfully regrows from this disuse atrophy as old muscle had no regrowth after 77 days of recovery. Hypothesis 2 reads: \"Ten of the 200 mRNAs corresponding to growth factors, growth factor receptors, or post-receptor signaling that are present on the employed microarray will differ between young and old rats during the failure of old skeletal muscle to rescue itself from immobilization-induced atrophy during reloading. Specific aim 1 will identify a pool of mRNAs associated with the failure of old skeletal muscle to rescue itself from immobilization-induced atrophy during reloading. Another observation is that short- term IGF-I application will rescue the failure of old muscle to regrow after disuse atrophy. However, this effect is only transient as continued IGF-I application to old muscle depletes remaining satellite cell proliferations and muscle wastes. Thus, all

Studies 31 defective growth factor responses must be identified. Specific aim 2 will apply IGF-I to old muscle after hindlimb immobilization in order to further identify those genes that failed to respond in old muscles, but had responded in young muscles. Specific aim 3 will begin to characterize for those differentially expressed ESTs between young and old muscles after ending immobilization in Specific aims 1 and 2. To support data analysis in Specific aims 1 and 2, Specific aim 4 will develop a computer-based, automated system for analysis of microarray data, data warehousing system for high capacity data storage, and tools for querying microarray data across experiments. Identifying the inappropriately expressed mRNAs associated with failed muscle regrowth of old muscles will permit a more scientifically-based growth factor rescue of old atrophied muscle. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: FUNCTIONAL DYNAMICS OF MOTOR CONTROL Principal Investigator & Institution: Brezina, Vladimir; Physiology and Biophysics; Mount Sinai School of Medicine of Nyu of New York University New York, Ny 10029 Timing: Fiscal Year 2002; Project Start 15-JUN-2001; Project End 31-MAY-2005 Summary: from applicant's abstract) The long-term goal of this research is to understand the basic computational and control principles which the central nervous system uses to generate functional behavior. Some fundamental principles are implicit in the interaction of the central controller with its peripheral effectors, most importantly muscles. The motor commands of the nervous system and the peripheral response characteristics of the neuromuscular system must be mutually matched for optimal performance. In many systems this matching is accomplished by peripheral modulation which dynamically tunes the properties of the muscle so as to enable it to perform the behavior being commanded by the nervous system. But, although set up as part of the behavior, the modulation generally has much slower dynamics than those of the behavior. In effect, the modulatory state represents a memory, maintained peripherally in the muscle, of past behavior. This memory then prepares the muscle to perform future behavior. It facilitates performance especially of the same kind of behavior as in the past, but may complicate performance if the nervous system commands a different behavior without its presence into account. This peripheral memory and its consequences for control of motor performance and behavior by the nervous system will be studied in a well known, experimentally advantageous model neuromuscular system. The system participates in several behaviors and exhibits a rich variety of neuromuscular modulation on a wide range of time scales. Preliminary studies demonstrate prominent peripheral memory in the system. A strategy combining experiments with mathematical modeling will be used to address the following questions: What motor commands does the nervous system send in the different behaviors? What corresponding modulation occurs? How do the commands and modulation interact to produce functional movement? How does the functional movement change when on the one hand the motor commands, and on the other hand the modulation, are altered? Altogether, this work will test a two-part hypothesis, reflecting the mutual interdependence of controller and effector: that the peripheral memory is required for smooth, efficient integration of successive cycles of a behavior and even for transitions from one behavior to another; but that, at the same time, its existence requires modification of the commands sent by the central nervous system. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

32 Muscles • Project Title: FUNCTIONAL RESPONSES OF EXTRAOCULAR EYE MUSCLES TO T3 Principal Investigator & Institution: Rubinstein, Neal A.; Associate Professor; Anatomy; University of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104 Timing: Fiscal Year 2002; Project Start 01-DEC-1997; Project End 31-MAR-2003 Summary: Thyroid dysfunction affects some 10 million Americans; and although the extraocular muscles (EOMs) are often involved in thyroid disease, little is known about the effects of T3 on the properties or development of EOM fibers. The effects of dysthyroidisms on the function of appendicular muscle fibers suggest that altered T3 levels should have profound influences on the performance of EOM fibers; however, there unique developmental origin, structural and functional properties and singular reactions to diseases suggest that EOMs have unique rules governing gene expression. T3 regulates the contractile properties of muscle fibers by differentially activating or repressing isoforms of the myosin heavy chains (MyHCs). The transcriptional control is mediated by the thyroid receptors (TRs) and the retinoid X receptors (RXRs) which themselves exist as multiple isoforms. Preliminary data, as well as susceptibilities to disease, suggest that the response of genes to T3 in EOMs will differ from that in other muscles. We hypothesize this differential response will be related to unusual distributions of TR and RXR isoforms among fibers; altered T3 levels will lead to the expression of inappropriate MyHC isoforms, abnormal contractile characteristics and impaired vision. Proving this hypothesis requires (a) determining which MyHC genes are expressed in each EOM fiber type during development and in the adult, (b) correlating the MyHC complement of each fiber to the contractile properties of that fiber, (c) determining whether hypo-and hyperthyroidism after the expression of MyHC genes and contractile properties, (d) discriminating the TR and RXR isoforms synthesized in euthyroid and pathological conditions. Studies will isoform-specific cRNA probes and antibodies will be combined with contractile measurements of individual skinned EOM fibers to accomplish these aims. To understand how the eye performs its repertoire of motions under both normal and pathological circumstances, one must understand the synthetic capacity of each fiber and how it defines the functional properties of each fiber. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: GENE THERAPY FOR A SEVERE DMD ANIMAL MODEL Principal Investigator & Institution: Xiao, Xiao; Associate Professor; Molecular Genetics & Biochem; University of Pittsburgh at Pittsburgh 350 Thackeray Hall Pittsburgh, Pa 15260 Timing: Fiscal Year 2004; Project Start 15-APR-2004; Project End 31-MAR-2009 Summary: (provided by applicant): Duchenne muscular dystrophy (DMD), caused by mutations in the dystrophin gene, is the most common, disabling and lethal muscle disease, afflicting one of every 3500 males. Recently, we have generated a series of highly truncated mini-dystrophin genes that had large deletions in the \"non-essential\" regions including part of the central rod domain and the very C-terminus domain. These minigenes were small enough to be packaged into adeno-associated virus (AAV) vectors and large enough to preserve high functionality, when tested in mdx mice after local intramuscular gene delivery. However, the mdx mice are far from an ideal DMD animal model although it is a commonly used one. While manifesting many similar symptoms of the human patients such as the muscle pathology, the mdx mice do not suffer shortened lifespan and do not show overall muscle weakness and skeletal contractures

Studies 33 as do the human patients. This phenomenon is due to the up-regulation of utrophin gene (a dystrophin analogue) that partially compensates the defects of dystrophin in the mdx mice. By knocking out both dystrophin and utrophin genes (double-KO), two teams have recently developed a severe DMD mouse model that closely reflects every major deficiency seen in the human patients including much shortened life-span, severe muscle weakness and skeletal contractures, offering a more truthful small animal model for more stringent tests of new therapeutics. In this grant proposal, we will use the newly available double-KO mice to vigorously test the hypothesis whether the novel mini-dystrophin genes are able to rescue the muscle functions locally and systemically, and more importantly, to improve the overall health and prolong the life-span of the severe DMD animal, which is key to the development of a clinically efficacious gene therapy strategy. In this proposal, we will investigate 1) biological/therapeutic functions of mini-dystrophin genes in the double-KO mice using the transgenic mouse technology; 2) therapeutic effects of mini-dystrophin genes in both young and adult double-KO mice after local intramuscular injection of AAV vectors; 3) systemic gene delivery and its therapeutic effects in large groups of muscles and the entire body; 4) alternative therapeutic genes that may offer synergistic effects along with the minigenes to benefit the dystrophic muscles. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: GENETICS AND MOLECULAR BIOLOGY OF MYOSIN Principal Investigator & Institution: Bernstein, Sanford I.; Biology; San Diego State University 5250 Campanile Dr San Diego, Ca 92182 Timing: Fiscal Year 2002; Project Start 01-JUL-1983; Project End 31-MAR-2004 Summary: (adapted from investigator's abstract): The investigators are using an integrative and multidisciplinary approach to determine how the myosin heavy chain (MHC) protein drives muscle function. Myosin is the molecular motor of muscle and the major component of myofibrillar thick filaments. Its ATP-dependent interaction with actin-containing thin filaments powers muscle contraction. They will test a series of hypotheses that predict myosin properties encoded by alternative exons, and how these properties dictate the different mechanical functions of various muscles. They use the model organism Drosophila melanogaster because it has a single gene coding for muscle MHC, but produces multiple forms of the protein (isoforms) by alternative RNA splicing. Using MHC null mutants in conjunction with germline transformation, they created a series of \"isoform-switch\" organisms that accumulate versions of MHC differing in single domains. To determine how each alternative structural domain defines the biochemical and biophysical properties of myosin and the ultrastructural and physiological properties of muscle, they are employing a battery of in vitro and in vivo assays: ATPase, actin and nucleotide binding, in vitro motility, optical trapping, electron microscopy, whole organism muscle function and isolated fiber mechanics. As appropriate, they will create a second series of chimeric constructs, to more specifically link functional properties with structural subdomains within each alternative region of the myosin head. Defining whether in vitro properties dictate in vivo functions is difficult, since a biochemical activity of a protein may always correlate with a particular mechanical property of a muscle without there being a causal relationship. The Drosophila muscle system is unique in that the effects of individual functional domains can be tested in muscle cells and intact organisms. Therefore, they can determine directly and to what degree a specific biochemical property defines a mechanical characteristic. They will also use these assays to test hypotheses regarding the molecular, biochemical, physiological and ultrastructural defects associated with two

34 Muscles Mhc mutations that affect key amino acid residues. Their results will be interpreted in relation to the three-dimensional structure of the myosin molecule and models for the mechanochemical cycle. Overall, their novel approach will yield direct insight into how the myosin protein functions in muscle and permit testing of models for the transduction of chemical energy into movement. Since mutations in the myosin head cause defects in human cardiac and skeletal muscle, these studies are relevant to understanding human myopathies. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: GTPASE REGULATION OF SMOOTH MUSCLE CONTRACTION Principal Investigator & Institution: De Lanerolle, Primal; Professor; Physiology and Biophysics; University of Illinois at Chicago 1737 West Polk Street Chicago, Il 60612 Timing: Fiscal Year 2002; Project Start 01-JUL-2001; Project End 31-MAY-2005 Summary: Small GTP binding proteins transduce signals that control a host of cellular responses. The activation of protein kinases by the GTP-bound form of small G proteins and highly regulated changes in the actin cytoskeleton appear to be important characteristics of the signalling properties of specific GTPases. Myosin II is an actin based molecular motor that converts chemical energy into mechanical work. The actin- myosin II interaction in smooth muscles is regulated by the phosphorylation of ser 19 on the 20 kDa light chain of smooth muscle myosin by the calcium-calmodulin dependent enzyme myosin light chain kinase. Recent reports have suggested that GTPases and myosin phosphorylation interact to regulate the actin cytoskeleton and smooth muscle contraction. Experiments performed by Somlyo, Kaibuchi and Narumiya and their co- workers have shown that GTPases regulate the calcium sensitivity of smooth muscle contraction. We have recently discovered that myosin light chain kinase is regulated by phosphorylation by PAK 1, a member of a family of protein kinases that is activated by the binding of the p21 GTPase. Although PAKs are generally thought to be involved in responding to stress, substrates for PAK's are not well characterized and myosin light chain kinase represents an important one. Based on extensive preliminary data, we propose that PAKs regulate the calcium sensitivity of smooth muscle contraction by a mechanism that involves the phosphorylation of myosin light chain kinase. We now propose experiments to test this hypothesis. The experiments described in Specific Aim 1 will investigate the kinetics of myosin light chain kinase phosphorylation by PAK 1, in vitro. Specific Aim 2 will test the hypothesis by performing studies on skinned smooth muscles. Specific Aim 3 will test the hypothesis by performing experiments on intact blood vessels. These experiments represent a powerful test of our hypothesis and they will provide important insights into the molecular mechanisms that regulate the calcium sensitivity of smooth muscle contraction. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: IGF-1 GENE TRANSFER TO ACCELERATE MUSCLE RECOVERY Principal Investigator & Institution: Vandenborne, Krista H.; Chair and Associate Professor; Physical Therapy; University of Florida Gainesville, Fl 32611 Timing: Fiscal Year 2002; Project Start 01-FEB-2002; Project End 31-JAN-2006 Summary: Muscle weakness is a common clinical phenomina observed following bed rest, surgery, cast immobilization and injury or disease. The consequences of loss of muscle strength are far reaching and include decrease of motor control and overall fitness, development of functional limitations and impairment, and long term disability. As such, the objective of this study is to investigate the potential of virus-mediated gene

Studies 35 transfer of IGF-1 to guard skeletal muscle from the deleterious impact of disuse or forced inactivity and to accelerate the subsequent recovery in muscle size and strength. For this purpose the left or right hindlimb (randomized) muscles of young adult mice will be injected with a recombinant adeno-associated virus vector for IGF-1. 3 months post-injection, both hindlimbs (injected and control) of the animals will be immobilized in a plaster cast for a period of 2 weeks. After removal of the cast the animals are allowed to reambulate and resume their normal cage activity. Cage restricted levels of weight-bearing activity have been shown to be sufficient to induce muscle regeneration and hypertrophy. Morphometric and functional measurements will be performed bilaterally (injected and control limb) at baseline, 3 months post-injection, following 2 weeks of cast immobilization, during reloading and at several time points during reambulation (2, 4 and 10 weeks). Morphological measures will include fiber cross- sectional area and fiber number, wet weight and protein content. Functional measures (twitch and tetanic force) will be performed in vitro on superfused muscles. The secondary objective of this study is to elucidate the mechanisms by which IGF-1 overexpression modulates muscle size and function under varying loading/activity conditions. For this purpose we will measure IGF-1 peptide levels, in vivo protein synthesis and degradation rates, and markers of muscle regeneration and satellite cell proliferation. We anticipate that the ability to locally manipulate muscle regeneration and hypertrophy during disuse and subsequent rehabilitation will be of great clinical importance. In addition, we anticipate that this study will help elucidate the role of IGF- 1 in the regulation of muscle size under varying loading/activation conditions. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: INJECTABLE SENSORS FOR CONTROL OF FES Principal Investigator & Institution: Loeb, Gerald E.; Professor and Director; Biomedical Engineering; University of Southern California 2250 Alcazar Street, Csc-219 Los Angeles, Ca 90033 Timing: Fiscal Year 2002; Project Start 10-JUN-2000; Project End 31-MAY-2004 Summary: (adapted from the Investigator's abstract): In order to reanimate a paralyzed limb to produce clinically useful movements, three functions must be provided: 1) Electrical stimulators to cause muscles to contract; 2) A controller to coordinate the stimulation; and 3) Sensors of command and feedback signals from the patient to the controller. The investigators have recently completed development and preclinical testing of a novel stimulation technology that permits large numbers of individual muscles to be precisely controlled by injectable, wireless microstimulators that receive power and data by RF transmission from an external controller. They propose to extend that technology by incorporating and testing various types of sensors in similar injectable modules. These will use a novel, compatible system for RF back-telemetry to send signals out of the limb for command and feedback purposes. Their immediate goal is a family of generic \"BIONs\" (bionic neurons)-that can be configured flexibly to serve a wide range of Functional Electrical Stimulation (FES) applications. The investigators have selected the following basic sensing modalities: 1) Low-level bioelectric signal recording such as electromyography, to monitor (the) level of electrical recruitment (M- waves) and spontaneous activity from muscles with some remaining voluntary control (useful as myoelectric commands for prostheses);and 2) Triangulation of relative position between devices, to be used for determining limb posture Acceleration and inclination (vs. gravity), using microelectromachined silicon (MEMS) sensor technology. The research will proceed in overlapping stages, the first of which is already underway in pilot work: 1) Design, build and test the basic circuit functions for low-level, low-

36 Muscles power signal detection, digitization and telemetric transmission; 2) Design, build and test specialized MEMS sensors; 3) Build complete injectable BIONs with sensing and back-telemetry capabilities; 4) Perform preclinical tests of sensor BIONs for biocompatibility; 5) Test sensing and telemetry functions in vitro with artificially- generated inputs; and 6) Test sensing and telemetry functions in alert, behaving animals. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: INSTABILITY AND MUSCULAR DEMAND DURING OBSTACLE CROSSING Principal Investigator & Institution: Chou, Li-Shan; Exercise & Movement Science; University of Oregon Eugene, or 97403 Timing: Fiscal Year 2003; Project Start 04-APR-2003; Project End 31-MAR-2005 Summary: (provided by applicant): Falls are among the most serious problems facing the aging population and have become the largest single cause of accidental death. Moreover, the total direct cost of fall injuries in 1994 among people 65 and older was $20.2 billion. Most falls in the elderly stem from interactions between environmental hazards and increased individual susceptibility to hazards from accumulated effects of age and intrinsic factors. Research on biomechanics of selected physical tasks, that take both environmental and intrinsic factors into account, is needed to quantify impairment magnitudes, to determine what elements are critical to the impairment, and ultimately to design more effective interventions for preventing falls in the elderly. The long-term goals of this proposed project are to advance the understanding of the mechanisms underlying the increased incidence of falls in the elderly, to determine a more effective method of identifying aged persons at risk of falling, and eventually to design more effective exercise/strengthening programs for the prevention of falls in the elderly. Specific aims of this project are to (1) demonstrate that motion of the whole body center of mass (COM) during obstacle crossing could better distinguish fallers from non-fallers when compared to individual segmental motion, (2) examine the relationship between ability to accommodate to environmental hazards during locomotion and muscle weakness, and (3) to identify quantitative, biomechanical indices (muscular demand-to- capacity ratios) that can better indicate the level of mechanical challenge imposed on selected muscles. Motion analysis and muscle strength testing will be performed on 24 elderly non-fallers and 24 elderly fallers (65 years or older). Body segment motion, ground reaction forces, and electromyography will be collected during unobstructed walking and stepping over obstacles of heights corresponding to 2.5% and 10% of each subject's height. Isometric strength of selected lower extremity muscles will be measured bilaterally. A thirteen-link biomechanical model, with kinematic inputs of each body segment and ground reaction forces will be used to compute the three- dimensional motion of the whole body COM and three-dimensional joint moments (torques) of the lower limbs. Data analysis will be performed on both mechanical and neuromuscular levels, including the isometric muscle strength, electromyography, motion of the COM, and it's interaction with the center of pressure (COP) of the stance foot derived from ground reaction forces and moments. Finally, correlation between muscle strength and dynamic balance control (indicated by the motion of the whole body COM) will be examined. This proposed project is expected to identify/define more sensitive biomechanical measures (both intrinsic and extrinsic) for better quantification of age-related mobility impairment and functional challenges imposed on our musculoskeletal system. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

Studies 37 • Project Title: INTERACTION AFFECTING CRANIOFACIAL MUSCLE DIFFERENTIATIO Principal Investigator & Institution: Noden, Drew M.; Professor; Biomedical Sciences; Cornell University Ithaca Office of Sponsored Programs Ithaca, Ny 14853 Timing: Fiscal Year 2002; Project Start 01-APR-2002; Project End 31-MAR-2005 Summary: (provided by applicant): Craniofacial muscles show great structural and functional diversity, and anomalies in their development contribute to a wide range of problems in vision, facial expression, mastication, and phonation. However, little is known about processes controlling the differentiation and maturation of branchial and extra-ocular muscles. Head muscles arise in mesenchyme adjacent to the brain then migrate and differentiate in connective tissues derived from the neural crest. Trunk and limb muscles arise in epithelial myotomes, then differentiate in paraxial and lateral mesoderm. This research will characterize tissue interactions necessary (1) to promote early differentiation of branchial and extra-ocular muscle myoblasts and (2) to direct expression of fiber typespecific myosins in primary myotubes in these muscles. These interactions will be identified by transplanting specific muscle precursors, at different stages and with or without putative muscle-inducing tissues, between head and trunk axial levels. Assays of tissue response include expression of myoblast markers (e.g., my, about, myoD), of trunk or head muscle-specific markers (e.g., parc aboutcis, pax3; podl, barx2, Ibxl,en2), and at later stages of muscle-specific myosin heavy chains. Surgeries are done on quail and chick embryos, which are ideally suited for in vivo tissue recombination experiments and for which detailed cellular and molecular biographies of head muscles are uniquely available. A third specific aim will characterize the processes by which craniofacial muscles change their attachments during later growth and maturation stages. EM studies reveal that primary myotubes undergo focal degeneration at their myotendinous tips and also at mid-myotube regions during embryonic days 12-15. I propose that these focal degenerations are essential for normal remodeling of head muscles, but nothing is known of the processes or provocations for these focal losses. Assays for elements of the pathways known to be active during programmed cell death of mononucleated cells will be applied to normal and function- or growth-arrested head muscles. Collectively, these experiments will provide the first insights into the signals necessary for initiation and diverse differentiations of craniofacial muscles and the mechanisms by which remodeling of these head muscles occurs. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: ISCHEMIC MITRAL REGURGITATION:FROM MECHANISMS TO THERAPY Principal Investigator & Institution: Levine, Robert A.; Professor; Massachusetts General Hospital 55 Fruit St Boston, Ma 02114 Timing: Fiscal Year 2002; Project Start 10-JUL-2001; Project End 31-MAY-2006 Summary: (provided by applicant) Immediate Goals: To examine the mechanism of ischemic mitral regurgitation (MR) with the goal of designing and implementing more effective therapy to reduce adverse impact on patients. Career Development Goals: To provide sufficient time for mentoring and research activities. Research Project: Mitral valve function can be understood in terms of the force-balance concept in which tethering forces from the papillary muscles balance left ventricular valve closing forces. In ischemic MR, this force balance may be altered in ways that impair the ability of the mitral leaflets to close effectively at the annular level. This proposal uses a combined,

38 Muscles parallel clinical and experimental approach to evaluate the mechanism, progression and therapy of ischemic MR, all relating to the central hypothesis that ischemic MR is caused by an abnormal relationship of the mitral valve to its supporting ventricular structures. These altered relationships involve both abnormal tethering forces due to displacement of the papillary muscles as well as reduced closing forces due to LV contractile dysfunction. Specific testable questions related to this hypothesis include: 1) The progression of mitral regurgitation in patients with acute myocardial infarction relates to abnormalities in the mitral valve-ventricular relationship; 2) These mechanisms also cause persistent MR despite coronary revascularization surgery, thereby impairing exercise capacity and raising pulmonary pressures; 3) Both an externally applied device and afterload reduction provide effective means of reducing ischemic mitral regurgitation by normalizing these relationships between the valve and the ventricle; cutting a minimum number of critically positioned strut chordae also has the potential to relieve tethering, and opens the way to potential minimally invasive percutaneous approaches. The aims of the mentored award will be met by allowing the PI to translate his experimental expertise to direct clinical studies of progression and functional outcome of ischemic MR, and to make the transition from mechanism to therapy in models reflecting the clinical situation, with the ultimate goal of patient applications. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: LOCOMOTOR DYNAMICS OF MUSCLE FUNCTION Principal Investigator & Institution: Biewener, Andrew A.; Professor & Chair; Organismic & Evolutionary Biol; Harvard University Holyoke Center 727 Cambridge, Ma 02138 Timing: Fiscal Year 2003; Project Start 01-JUN-2003; Project End 31-MAY-2008 Summary: (provided by applicant): The proposed research addresses the central question of how muscles function under dynamic conditions of locomotor activity. It does so in the context of how muscle function is modulated in relation to muscle architecture and fiber composition to accommodate changes in locomotor requirement. These questions will be addressed by making in vivo recordings of force (tendon buckle transducers), length change (sonomicrometry) and neural activation (electromyography) of key limb muscles in two animal models: quadrupedal goats and bipedal guinea fowl. Measurements will be obtained from animals trained to move over a range of speeds on a treadmill at different gaits and grades (level vs incline vs decline) to address the following hypotheses: (i) regional activation and fractional length change within muscles that have focal skeletal attachments is uniform both along a fascicle axis and between differing fascicle regions, but may vary in muscles with broader attachments and more complex architectures; as a result, (ii) the timing and strain of activated fascicles are homogeneous within a muscle performing a given motor task; and (iii) proximal muscles with long fibers account for the majority of mechanical work modulation; whereas distal short-fibered muscles with long tendons contract isometrically for more economical force production and tendon elastic savings. Differences in mechanical work rate with locomotor grade will be related to observed changes in the in vivo force-length behavior of key limb muscles. Recordings made while animals accelerate from rest will provide a second context to evaluate work modulation in relation to muscle architecture. Ground reaction force-platform and high- speed video recordings will also be carried out to integrate the in vivo force, length and EMG measurements of individual muscles into whole-limb mechanics. These studies have important consequences for understanding patterns of motor recruitment in relation to locomotor strategy and how regional differences in motor unit organization

Studies 39 (and fiber type) may influence the neural control of movement. Prior work in this area has been limited by studies of motor function under more quasi-steady ranges of movement and/or indirect assessment of muscle length change and force development. Although an overarching goal is to understand factors that influence normal and age- related changes in human motor function, animal studies allow direct experimental approaches for assessing the dynamics of motor function that are likely to apply to humans. Consequently, the proposed studies will have value for developing more effective physical, occupational and rehabilitative therapies, as well as for sports and exercise training, and prosthetics design. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: LUNG GROWTH AT HIGH ALTITUDE AND MAXIMAL O2 TRANSPORT Principal Investigator & Institution: Johnson, Robert Lee.; Professor; Internal Medicine; University of Texas Sw Med Ctr/Dallas Dallas, Tx 753909105 Timing: Fiscal Year 2002; Project Start 01-JUN-1996; Project End 31-MAR-2006 Summary: (Applicant's abstract): Human natives of high altitude (HA) develop increased lung volume and diffusing capacity consistent with enhanced alveolar growth, and increase blood volumes that facilitate 02 transport. However, other adaptation to HA (muscularization of pulmonary arterioles, dysanaptic airway growth and retardation of thoracic growth) may impair O2 transport. The interplay among these factors at different altitudes is not known. Functional consequences of these structural changes can be isolated only after re-acclimatization to sea level (SL) when reversible changes in blood volume and pulmonary vascular reactivity have subsided. We employ this approach to address long-term structure-function relationships of maturation at HA in dogs. Hypotheses are 1) Hypoxia stimulates alveolar hyperplasia and enhances diffusive gas exchange. 2) Airway growth lags behind alveolar growth at HA, leading to uneven distribution of ventilation and increased ventilatory work. 3) Pulmonary vascular changes at HA significantly limits maximal cardiac output at SL. 4) Structural changes at HA persist after reacclimatization to SL and exert opposing effects on O2 transport, i.e., persistent vascular and airway abnormalities offset benefits derived from enhanced alveolar growth. 5) Growth of thorax is impaired in an altitude- dependent way; at extreme altitude, the restricted thoracic size sets an upper limit to lung growth and O2 transport. We plan to raise immature dogs (age 2 mo.) to somatic maturity (12 mo.) at 3 levels of HA (3, 100m, 3,800m or 4,500m in separate groups) compared with controls raised at SL. Dogs will be returned to SL at maturity for cardiopulmonary testing at rest and exercise, including pressure-volume curves, maximal 02 uptake, efficiency of gas exchange and diffusing capacity of lungs (DL) and muscles, ventilatory work, hemodynamics and blood volume. Dimensions of airways, diaphragm, rib cage, lungs and spleen will be assessed by spiral CT scan. Components of DL, septal tissue volume and pulmonary blood flow will be measured at regular intervals and correlated with blood volume. After 1 yr. of re-acclimatization to SL, studies will be repeated to determine regression of changes. Terminally, detailed structural analysis will be performed on the lungs, respiratory, locomotive and ventricular muscles, as well as ribs and long bones. Growth patterns of the acini, airways, vasculature, thoracic structures and their functional correlates will be compared at the 3 levels of hypoxia to determine the altitude-dependence of adaptation in O2 transport. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

40 Muscles • Project Title: MECHANICAL FUNCTION OF MUSCLE DURING MOVEMENT Principal Investigator & Institution: Marsh, Richard L.; Professor; Biology; Northeastern University 360 Huntington Ave Boston, Ma 02115 Timing: Fiscal Year 2002; Project Start 01-FEB-2001; Project End 31-JAN-2006 Summary: The performance of a skeletal muscle during movement is determined by the interaction of its intrinsic properties with the mechanical properties of the system to which it is linked. These interactions are complex and predictive equations are limited in some respects by the lack of empirical data on the performance of muscle under loading conditions that replicate those found during movement. Muscles serve three major mechanical functions during movement: producing force, producing work, and providing stability. These different functions are linked to differences in the length trajectory (sequence of length change) in relation to the motor activity of the muscles. Although we know that all three of these functions are performed by humans in walking and running, we know little of the conditions under which individual muscles operate while performing each function. Further we have no empirical data on the quantitative importance of each function to the cost of locomotion. The specific aims of this project are to: 1) Examine the in vivo contractile parameters (operational lengths, length trajectories, and amounts of series elasticity) for muscles that are active only while performing positive work in running and jumping; 2) Examine the prediction that during running and jumping actively lengthening muscles function to help stabilize the movement; 3) Quantify the relative energetic importance of the different mechanical functions served by muscles during running; 4) Measure the efficiency of fast and slow muscles under conditions of varying power output; 5) Quantify the influence of velocity dependent activation and deactivation on mechanical function of fast and slow muscles. The mechanical function of muscles used in running and jumping will be assessed in vivo using sonomicrometry and electromyography. Blood flow measurements using fluorescently labeled microspheres will be used in conjunction with other measures to estimate the relative contribution of the different limb muscles to the energy cost of running. In vitro work with the muscles used in jumping and running and computer modeling will examine the optimum conditions for accelerating inertial loads. Mouse muscles will be used to examine the influence of length trajectory and cycle frequency on mechanical performance and efficiency. This project is predicated on the assumption that examining how muscles are used in animals during movement allows us to better predict the design parameters important in human movement and will improve our understanding of both normal and dysfunctional human movement. Such studies will eventually assist in designing rehabilitative strategies that require an understanding of the diverse roles of muscles during movement. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: MECHANICAL VENTILATION AND RESPIRATORY MUSCLES Principal Investigator & Institution: Powers, Scott K.; Professor and Chair; Exercise and Sport Sciences; University of Florida Gainesville, Fl 32611 Timing: Fiscal Year 2002; Project Start 01-FEB-2001; Project End 31-JAN-2005 Summary: (Applicant's abstract): Mechanical ventilation (MV) is used clinically to sustain ventilation in patients who are incapable of independently maintaining adequate alveolar ventilation. Unfortunately, the withdrawal of MV, or weaning, can be difficult in a large number of cases. Strong evidence exists that MV-induced respiratory muscle weakness contributes significantly to these difficulties in weaning. Indeed, we have recently demonstrated that prolonged MV results in diaphragmatic atrophy and a

Studies 41 significant reduction in diaphragmatic maximal force production. Further, we have observed that prolonged MV results in oxidative injury (i.e. protein oxidation) to the diaphragm; this is significant because oxidized proteins become targets for proteases. The mechanisms responsible for this MV-induced atrophy and protein oxidation are unknown and comprise the focus of our proposed experiments. To determine the factors that contribute to diaphragmatic atrophy during prolonged MV, we will test the following hypotheses: 1a) MV-induced diaphragmatic atrophy occurs due to a decrease in synthesis of muscle proteins as well as an increased rate of proteolysis; 1b) proteolysis is the major contributor to diaphragmatic protein loss during prolonged MV; 2a) The increased activity of calpain, lysosomal, and ATP ubiquitin-dependent proteases are collectively responsible for the protein degradation observed in diaphragms from MV animals; and 2b) Although calpain, lysosomal, and ATP-ubiquitin-dependent proteases all contribute to diaphragmatic protein loss during MV, the ATP-ubiquitin- dependent and calpain proteolytic pathways are dominant. To resolve which chemical pathways are responsible for diaphragmatic protein oxidation during MV we will test the hypothesis that MV-induced protein oxidation in the diaphragm is caused by several reactive chemical species including hypochlorous acid, tyrosyl radicals and hydroxyl radicals. To test these postulates, we will perform both in vitro and in vivo studies using an animal model and utilize the tools of molecular biology, biochemistry, and physiology. These experiments will improve our understanding of the mechanisms associated with MV-induced diaphragmatic atrophy. The long-term goal of our experiments is to provide the knowledge required to develop clinical strategies to oppose the deleterious effects of MV on respiratory muscles. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: MECHANISMS OF SEXUAL DIFFERENTIATION IN NEURAL SYSTEMS Principal Investigator & Institution: Forger, Nancy G.; Professor; Psychology; University of Massachusetts Amherst 408 Goodell Building Amherst, Ma 01003 Timing: Fiscal Year 2002; Project Start 12-JUN-2000; Project End 31-MAY-2005 Summary: (adapted from applicant's abstract): A large number of sex differences in the central nervous systems (CNS) of vertebrates have now been described. Such morphological dimorphisms may underlie well documented sex differences in behavior, in susceptibility to certain drugs, and in the incidence of some human mental disorders including autism, depression and schizophrenia. In many cases, neural sex differences have been shown to be due to differential exposure to gonadal steroid hormones in males and females. However, the cellular and molecular mechanisms governed by hormones in the nervous system are not well understood. The identification and cloning of several new neurotrophic molecules has fueled an explosion of research into the actions of trophic molecules in the CNS, and recent findings indicate a role for neurotrophic factors in sexually dimorphic development. Experiments in the first half of this proposal will test the idea that effects of gonadal steroids are mediated by trophic factors in a well-characterized model system. The spinal nucleus of the bulbocavernosus (SNB) and its target muscles constitute an anatomically simple system that is sexually dimorphic in many mammals. SNB motoneurons reside in the lumbar spinal cord and innervate striated perineal muscles attached to the phallus. Androgens regulate SNB motoneuron survival during perinatal development, and SNB cell size in adulthood. Recent observations suggest that some effects of androgens on this system are mediated by protein neurotrophic factors. Trophic factor antagonists will be administered to developing and adult rats in order to identify endogenously produced factors

42 Muscles controlling SNB cell survival and morphological plasticity. In the second half of this proposal, the intracellular events regulated by hormones and neurotrophic factors will be explored. Specifically, a role for the death-regulatory protein, Bcl-2, in sexually dimorphic cell death will be tested in the SNB and in the anteroventral periventricular nucleus (AVPV) of the hypothalamus. Because the neurotrophic factors and death- regulatory proteins to be examined are expressed throughout the nervous systems of many vertebrates, including humans, information gained from this work will be relevant to our overall understanding of the extracellular and intracellular molecules mediating hormone regulated development and plasticity in neural tissues. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen • Project Title: MENTAL EFFORT EFFECT ON LARGE MUSCLE STRENGTHENING Principal Investigator & Institution: Yue, Guang H.; Associate Staff; Cleveland Clinic Foundation 9500 Euclid Ave Cleveland, Oh 44195 Timing: Fiscal Year 2002; Project Start 27-SEP-1999; Project End 31-MAY-2004 Summary: It is well known that to strengthen a muscle one should perform training involving heavy loads or resistance. Recently we have found that substantial voluntary strength gains can be achieved with training involving low resistance but strong mental effort. In contrast, individuals who trained with the same low-intensity contractions but with low mental effort had no improvement in strength. Based on these preliminary findings, we hypothesize that muscle strength improvements depend primarily on the level of mental effort during training, not the training intensity (resistance) per se. The reason that high-intensity training always increases strength is because mental effort is high during high-intensity muscle contractions. Aim 1 of the project is to compare the effects of training with different levels of mental effort on the improvement in muscle strength. Four groups of elderly subjects (greater than or equal to 65 years) will participate in a 12-week training study involving elbow-flexor muscles. One group will be trained with an intensity near the level of maximal voluntary contraction (MVC group); a second group will be trained with high mental-effort, low muscle-intensity elbow-flexion contractions (LME group); and the fourth (control) group will not be trained will participate in the strength tests. We expect that the strength improvement after training will be: MVC group > HME group > LME group = control group. We also expect that the strength increase in the MVC and HME groups will result in an improvement in daily living function. Aim 2 is determine the neural mechanisms underlying muscle strength improvements. We hypothesize that an increase in the central nervous system (CNS) drive is the primary mechanism that mediates strength improvements induced by low-intensity training (HME group). To evaluate the CNS drive, four measurements will be made using the same subjects and groups as in Ami 1: brain activation level examined by functional MRI (fMRI) and EEG-derived motor activity-related cortical potential (MRCP), surface EMG signals, and the MRI T2 relaxation time obtained from the trained muscles. We expect to find that after training: (1) the brain activation level (fMRI and MRCP), EMG, and MRI T2 will significantly increase in the MVC and HME groups; and (2) the amplitude of increases in these measurements will be: MVC group = HME group > LME group = control group. The knowledge gained from these studies will substantially advance the current understanding of mechanisms underlying human voluntary muscle strengthening and will have direct application in neuromuscular rehabilitation for older adults and individuals who are physically handicapped and unable to perform repeated, forceful muscle contractions. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen