Partial or Complete Unloading of Skeletal Muscle Leads to Specific Alterations of Anabolic Signal Transduction

Consequences of disuse atrophy of skeletal muscle observed during spaceflight on astronaut health and performance are a focal point of space research. Decrements of both muscle mass and protein synthesis rates have been observed with exposure to varying muscle loading environments (1G \u3e partial loading \u3e 0G), and most of the reduced muscle mass can be attributed to diminished rates of synthesis. However, specific mechanisms behind unloading-dependent reductions of protein synthesis are not well defined. PURPOSE: To determine whether or not alterations of anabolic signal transduction was responsible for the changes previously observed in fractional synthesis rates with specific gravitational loading paradigms. METHODS: Female BALB/cByJ were normalized by bodyweight and assigned to normal cage ambulation (1G), partial weight bearing suspension titrated to approximately 33% bodyweight (G/3), partial weight bearing titrated to 16% bodyweight (G/6) and full unloading of hind limbs (0G) in specially designed cages. All mice were subjected to that loading environment for 21d prior to tissue harvest, and monitored daily. Immunoblotting of the gastrocnemius (n=23) was carried out to analyze alterations of anabolic signal transduction. Although numerous signaling intermediates were assessed, the focus of this abstract will be on ribosomal protein S6 kinase (p70-S6K). This important protein has served as a marker of protein synthesis signal transduction as well as the anabolic capacity in skeletal muscle. RESULTS: Regardless of loading paradigm, no differences were detected among groups for the activation of p70-S6K (as indicated by the phospho: total protein content). Total protein content, however, was ~27% lower than control in 0G and G/3 (P=0.008) with G/6 not being different from control (P\u3e0.05). CONCLUSION: In combination with previous data (unpublished observations), Partial gravitational fields at least partially rescues anabolic signaling, suggesting that a threshold level of stimulus is necessary to maintain anabolic capacity in muscle. These results may have important implications towards the development of strategies designed to counter the effects of partial/complete unloading on skeletal muscle based on how the anabolic capacity of muscle is affected

Autophagy is Required for the Anabolic Response to Muscle Contraction

Exercise is a key stimulus in regulating the behavior and metabolism of skeletal muscle, with exercise inducing muscular growth through activation of the anabolic mechanistic target of rapamycin kinase (mTOR). Separately, there is mounting evidence that exercise increases autophagy (one of the main routes by which intracellular proteins are degraded) and that the autophagic process may indeed be required for adaptations to exercise training. PURPOSE: To investigate the effects of autophagy inhibition on mTOR signaling and cellular anabolism after muscular contraction. METHODS: Cultured L6 myotubes were to exposed to electrical pulse stimulation using a stimulator set to deliver bipolar pulses of 30V at 100 Hz for 200 ms every fifth second for 60 minutes. Subsequently, cells received either vehicle control, or 100 μM NSC-185058, an antagonist of the key autophagy protein ATG4B and known inhibitor of autophagy. All groups were also exposed to 4% deuterium oxide, a stable isotopic tracer for measurements of protein synthesis. 24 hours post “exercise” bout, cells were lysed in ice-cold Norris buffer, and prepared for Western immunoblot of protein expression, or determination of protein fractional synthesis rate (FSR) of the myofibrillar fraction via mass-spectrometry analysis. Non-stimulated cells receiving vehicle control treatment served as controls, with a one-way analysis of variance and Tukey’s post-hoc test used to test for any differences between groups. RESULTS: We found that phosphorylation of a key downstream target of mTOR, P70S6 kinase, was roughly seven times greater in cells subjected to EPS and vehicle control (710.3%) relative to control (p0.05). While there was a trend for EPS treatment to increase expression of ATG4B, along with a reduction of ATG4B content as a result of NSC-185058 treatment, this finding did not rise to the level of statistical significance. There were no differences in FSR between cells exposed to EPS; however, NSC-185058 treatment significantly reduced FSR in EPS treated cells relative to controls (0.8712 %/hr vs 1.193 %/hr). CONCLUSION: These findings present two conclusions: high-intensity EPS as an in vitro model of exercise elevates mTOR signaling through P70S6K 24 hours post exercise, and mTOR activation as a result of muscular contraction is reliant upon autophagy in skeletal muscle. Further work will be required to elucidate the dynamics of this relationship, and the interplay between skeletal muscle autophagy and anabolism

Female mice may have exacerbated catabolic signalling response compared to male mice during development and progression of disuse atrophy

Background: Muscle atrophy is a common pathology associated with disuse, such as prolonged bed rest or spaceflight, and is associated with detrimental health outcomes. There is emerging evidence that disuse atrophy may differentially affect males and females. Cellular mechanisms contributing to the development and progression of disuse remain elusive, particularly protein turnover cascades. The purpose of this study was to investigate the initial development and progression of disuse muscle atrophy in male and female mice using the well-established model of hindlimb unloading (HU). Methods: One hundred C57BL/6J mice (50 male and 50 female) were hindlimb suspended for 0 (control), 24, 48, 72, or 168 h to induce disuse atrophy (10 animals per group). At designated time points, animals were euthanized, and tissues (extensor digitorum longus, gastrocnemius, and soleus for mRNA analysis, gastrocnemius and extensor digitorum longus for protein synthesis rates, and tibialis anterior for histology) were collected for analysis of protein turnover mechanisms (protein anabolism and catabolism). Results: Both males and females lost ~30% of tibialis anterior cross-sectional area after 168 h of disuse. Males had no statistical difference in MHCIIB fibre area, whereas unloaded females had ~33% lower MHCIIB cross-sectional area by 168 h of unloading. Both males and females had lower fractional protein synthesis rates (FSRs) within 24-48 h of HU, and females appeared to have a greater reduction compared with males within 24 h of HU (~23% lower FSRs in males vs. 40% lower FSRs in females). Males and females exhibited differential patterns and responses in multiple markers of protein anabolism, catabolism, and myogenic capacity during the development and progression of disuse atrophy. Specifically, females had greater mRNA inductions of catabolic factors Ubc and Gadd45a (~4-fold greater content in females compared with ~2-fold greater content in males) and greater inductions of anabolic inhibitors Redd1 and Deptor with disuse across multiple muscle tissues exhibiting different fibre phenotypes. Conclusions: These results suggest that the aetiology of disuse muscle atrophy is more complicated and nuanced than previously thought, with different responses based on muscle phenotypes and between males and females, with females having greater inductions of atrophic markers early in the development of disuse atrophy

Solar Occultation Satellite Data and Derived Meteorological Products: Sampling Issues and Comparisons with Aura MLS

Derived Meteorological Products (DMPs, including potential temperature (theta), potential vorticity, equivalent latitude (EqL), horizontal winds and tropopause locations) have been produced for the locations and times of measurements by several solar occultation (SO) instruments and the Aura Microwave Limb Sounder (MLS). DMPs are calculated from several meteorological analyses for the Atmospheric Chemistry Experiment-Fourier Transform Spectrometer, Stratospheric Aerosol and Gas Experiment II and III, Halogen Occultation Experiment, and Polar Ozone and Aerosol Measurement II and III SO instruments and MLS. Time-series comparisons of MLS version 1.5 and SO data using DMPs show good qualitative agreement in time evolution of O3, N2O, H20, CO, HNO3, HCl and temperature; quantitative agreement is good in most cases. EqL-coordinate comparisons of MLS version 2.2 and SO data show good quantitative agreement throughout the stratosphere for most of these species, with significant biases for a few species in localized regions. Comparisons in EqL coordinates of MLS and SO data, and of SO data with geographically coincident MLS data provide insight into where and how sampling effects are important in interpretation of the sparse SO data, thus assisting in fully utilizing the SO data in scientific studies and comparisons with other sparse datasets. The DMPs are valuable for scientific studies and to facilitate validation of non-coincident measurements

Arizona State Museum Archaeological Series No. 177

The 1979-1983 Testing at Los Morteros (AZ AA:12:57 ASM), A Large Hohokam Village Site in the Tucson Basin by Richard C. Lange and William L. Deaver. Contributions by Susan L. Brantley, Laverne Conway, T. Michael Fink, Paul R. Fish, Richard J. Harrington, James P. Holmlund, Felipe C. Jacome, Susan J. Wells, Sharon F. Urban. Compiled by Richard C. Lange. Archaeology Section, Research Division, Arizona State Museum, University of Arizona, 1989. Archaeological Series 177.Los Morteros (AZ AA:12:57 [ASM]) is a large Hohokam village site located at the northern end of the Tucson Mountains along the Santa Cruz River. Named for the bedrock mortars located near the center of the site, Los Morteros has a rich and varied history. Several phases of prehistoric occupation are suggested by ceramics representing the Colonial, Sedentary, and Classic periods (A.D. 500 to 1450). The range of features present is considerable, including cremation pits, pit houses, roasting pits, mounds, canals, petroglyphs, hill-side terraces (trincheras), a ballcourt, and the bedrock mortars. Spatially, th~ site covers a large area, but most features are clustered in a limited core area (Fig. 1.1). The area around Los Morteros has also been used during historical times (see Stein 1982). In this report reference to the historical period is limited to those events that bear directly on the site of Los Morteros, in particular the history of archaeological interest in the site. Our major concern is with the prehistoric components at Los Morteros. (excerpt from Introduction)Preface / Paul R. Fish, Richard C. Lange -- Introduction / Richard C. Lange -- Methods / Richard C. Lange -- Surface distribution of ceramic artifacts / William L. Deaver -- Pottery and other ceramic artifacts / William L. Deaver -- Feature descriptions / Richard C. Lange -- Specific testing activities / Richard C. Lange -- Discussion and summary / Richard C. Lange -- Appendix A. Trench details -- Apprendix B. Ceramic data -- Appendix C. Lithic data -- Appendix D. Feature data -- Apprendix E. Synopsis of soil profiles / James P. Holmlund -- Appendix F. The utility of phosphate testing at Los Morteros / Susan J. Wells -- Apprendix G. Faunal materials / Laverne Conway -- Appendix H. Pollen samples -- Appendix I. Macrobotanical remains / assembled by Charles H. Miksicek -- Appendix J. Analysis of a human skeleton (burial 1) from Los Morteros / Richard J. Harrington -- Appendix K. The cremated human remains from Los Morteros / T. Michael Fink -- Appendix L. Shell from Los Morteros / Sharon F. Urban -- Appendix M. Archaeomagnetic sample results -- Appendix N. Radiocarbon sample results -- Appendix O. Photographs of Los Morteros artifacts -- Appendix P. Chemical analysis of pigments on sherds from Los Morteros / Susan L. Brantley -- Appendix Q. Hohokam projectile points and their localities of manufacture in the Tucson Basin / Felipe Carlos Jacome.This title from the ASM Archaeological Series is made available by the Arizona State Museum and University of Arizona Libraries. If you have questions about this title, please contact Jannelle Weakly at the Arizona State Museum, (520) 621-6311, [email protected]

OUTDOOR RECREATION IN THE UNITED STATES WITH SPECIAL REFERENCE TO RECREATION AT U.S. ARMY CORP OF ENGINEERS SITES

Resource /Energy Economics and Policy,

Arizona State Museum Archaeological Series No. 147, Vol. 2, Part 2

Hohokam Habitation Sites in the Northern Santa Rita Mountains by Alan Ferg, Kenneth C. Rozen, William L. Deaver, Martyn D. Tagg, David A. Phillips, Jr., David A. Gregory, with contributions by Margaret Glass, Robert S. Thompson, Kurt Dongoske, Karl J. Reinhard, Richard H. Hevly, Richard C. Lange, Bruce B. Huckell. Submitted by Cultural Resource Management Division, Arizona State Museum, University of Arizona. Prepared for ANAMAX Mining Company, 1984. Archaeological Series No. 147, Vol. 2, Part 2.Comparisons show them to be similar to forms known from other Hohokam sites. In the ninth chapter an analysis of the factors influencing settlement location is presented. Variables such as topography, soils, vegetation, elevation, and distance to permanent water are found to have low correlations to site location, but location of sites is highly correlated to stream profile gradient. Comparisons with sites located in similar areas in southeastern Arizona suggest that this variable may have considerable explanatory and predictive power. The last chapter pulls together all available data to examine the nature of the Hohokam occupation of the Rosemont area. Functional site types and intrasite organization are first discussed. Three categories of functional site types (new farmsteads, stable or growing farmsteads, and one site with a ballcourt) are recognized, as is a pattern on intrasite organization which is probably based in part on Hohokam customs and in part on local topography. Economy and subsistence are next examined, and from the meager data available it is proposed that maize agriculture supplemented by hunting and gathering of upland fauna and flora supported the area's inhabitants. Examination of areal and regional relationships indicates principal contacts with the Phoenix Basin Hohokam, the Mogollon, and the Trincheras cultures. Intrusive decorated pottery is the primary evidence for contact, although the presence of a large number of inhumations and certain aspects of architecture provide further documentation of intercultural contact. The nature of the Tucson Basin Hohokam occupation of the area is next traced in terms of site distribution, population distribution, and intersite organization. Possible organization of the Rosemont "local system" is discussed, and reconstructions of temporal trends in settlement and population numbers are presented. Unstable climatic conditions, in conjunction with overexploitation of the local resource base, is suggested to be the ultimate cause of abandonment of the Rosemont area.Digital version includes redaction statements in the document.This title from the ASM Archaeological Series is made available by the Arizona State Museum and University of Arizona Libraries. If you have questions about this title, please contact Jannelle Weakly at the Arizona State Museum, (520) 621-6311, [email protected]

Arizona State Museum Archaeological Series No. 147, Vol. 2, Part 1

Hohokam Habitation Sites in the Northern Santa Rita Mountains by Alan Ferg, Kenneth C. Rozen, William L. Deaver, Martyn D. Tagg, David A. Phillips, Jr., David A. Gregory, with contributions by Margaret Glass, Robert S. Thompson, Kurt Dongoske, Karl J. Reinhard, Richard H. Hevly, Richard C. Lange, Bruce B. Huckell. Submitted by Cultural Resource Management Division, Arizona State Museum, University of Arizona. Prepared for ANAMAX Mining Company, 1984. Archaeological Series No. 147, Vol. 2, Part 1.Excavations at 22 ceramic period sites in the Rosemont area of the northern Santa Rita Mountains, Coronado National Forest, are described. Investigations have e~tablished that these sites were occupied by the Tucson Basin Hohokam more or less continuously from approximately A.D. 500 until abandonment of the area at approximately A.D. 1200. The ceramic period prehistory of southeastern Arizona is outlined in the first chapter, and the research design for work at the Rosemont sites is presented in the second chapter. Next each site that received investigation is briefly described, including discussions of the features, artifacts, and length of occupation of each. This is followed by a series of six chapters which present the results of specialized analyses of various classes of material culture. The first of these discusses the pottery from the sites. Detailed definitions and descriptions of the plain, painted, and red ware pottery types are presented, and a model of decorative stylistic development of Tucson Basin painted pottery is introduced. Potential and temporal changes are explored as well; a few temporal trends are noted, but no functional differentiation was identified. The flaked stone artifacts are rigorously described, and both the implements and debitage are subjected to detailed technological, formal, and functional examinations. This study demonstrates that the assemblages display a great deal of consistency in all attributes, and that there is little evidence of technological, formal, or functional variability among the sites. Only projectile point styles exhibit change through time. The ground stone artifacts, divided into utilitarian and nonutilitarian forms, are treated in the next two chapters. A series of formal and functional classes is defined for both, and the various classes are described in detail. Possible functions are discussed, and each class is compared to similar classes from other sites and other areas. It is noted that the Rosemont ground stone artifacts are generally simple, unembellished forms made of local materials, and that the assemblages from all sites are quite similar. Shell artifacts are next described and discussed, and are again found to be relatively simple forms, probably entering the area as finished items. Comparisons show them to be similar to forms known from other Hohokam sites. In the ninth chapter an analysis of the factors influencing settlement location is presented. Variables such as topography, soils, vegetation, elevation, and distance to permanent water are found to have low correlations to site location, but location of sites is highly correlated to stream profile gradient. Comparisons with sites located in similar areas in southeastern Arizona suggest that this variable may have considerable explanatory and predictive power. The last chapter pulls together all available data to examine the nature of the Hohokam occupation of the Rosemont area. Functional site types and intrasite organization are first discussed. Three categories of functional site types (new farmsteads, stable or growing farmsteads, and one site with a ballcourt) are recognized, as is a pattern on intrasite organization which is probably based in part on Hohokam customs and in part on local topography. Economy and subsistence are next examined, and from the meager data available it is proposed that maize agriculture supplemented by hunting and gathering of upland fauna and flora supported the area's inhabitants. Examination of areal and regional relationships indicates principal contacts with the Phoenix Basin Hohokam, the Mogollon, and the Trincheras cultures. Intrusive decorated pottery is the primary evidence for contact, although the presence of a large number of inhumations and certain aspects of architecture provide further documentation of intercultural contact. The nature of the Tucson Basin Hohokam occupation of the area is next traced in terms of site distribution, population distribution, and intersite organization. Possible organization of the Rosemont "local system" is discussed, and reconstructions of temporal trends in settlement and population numbers are presented. Unstable climatic conditions, in conjunction with overexploitation of the local resource base, is suggested to be the ultimate cause of abandonment of the Rosemont area.Figures / Tables / Preface / Acknowledgments / Abstract / The Rosemont Study Area and Previous Research in Surrounding Areas / Research Design / Site Descriptions / Pottery / Flaked Stone / Utilitarian Ground Stone / Nonutilitarian Ground Stone, Crystals, and Minerals / Shell / Ceramic Period Settlement Patterns in the Rosemont Area: A Discussion / Discussion / Appendices / ReferencesThis title from the ASM Archaeological Series is made available by the Arizona State Museum and University of Arizona Libraries. If you have questions about this title, please contact Jannelle Weakly at the Arizona State Museum, (520) 621-6311, [email protected]