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further assays. To determine whether bear serum had a seasonal effect in response toapoptotic threat, MC3T3-E1 pre-osteoblasts were primed for 24 hours in seasonal bearserum, then serum starved for 6 hours. The caspase-3/7 response to 6 hours’ serumstarvation was graphed longitudinally with a smoothed average curve (Figure 5.2). Thecaspase-3/7 response began to decrease in serum collected approximately 10 daysbefore hibernation, and remained at a lower level throughout hibernation. In serumcollected just after hibernation, caspase-3/7 response immediately rose to levels higherthan those stimulated by prehibernation serum, suggesting a state of increasedosteoblast metabolism in post-hibernation. When samples were grouped by season andcompared by ANOVA, caspase-3/7 activity dropped from 5.66+0.23 relative units in fallto 3.82+0.20 in winter; values then rose to 7.89+0.41 in the spring (p
involved in the reduced caspase-3/7 response of cells primed in hibernation serum,experiments were repeated with H-89 and pertussis toxin. When the protein kinase A(PKA) pathway was inhibited with H-89, the percent decrease between cells inprehibernation versus hibernation serum was 39% in the vehicle sample (p
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EXPLORATION OF THE ROLE OF SERUM FA
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Table of contentsIndex of Figures .
Page 5 and 6:
Chapter Four - Attempts to find an
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Index of figures1.1. Simplified sch
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6.17. Longitudinal analysis of OPG
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6.49. Longitudinal analysis of ColI
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A.2. Complete list of BSALP correla
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List of abbreviationsα-MEM. Alpha-
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PCR. Polymerase chain reactionPer1
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G2/M checkpoint. The point in the c
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Species names13-lined ground squirr
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Chapter One - Background and signif
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1.3 Bone turnover is unbalanced dur
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pro-apoptosis regulator BAX. An imp
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low-dose administration of parathyr
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decrease in bone trabecular thickne
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experience periodic arousals in whi
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ears must have evolved a mechanism
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1.7.6 Hypothesis 2: OCN interacts w
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ain, heart, and femoral muscle of h
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Hypothesis 3a: Serum concentrations
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were released. Behavior indicating
Page 47 and 48: 2.3 ResultsSerum activity of the bo
Page 49 and 50: (p
Page 51 and 52: post-hibernation sampling in the be
Page 53 and 54: also unclear whether ucOCN may affe
Page 55 and 56: atios of intact to fragmented OCN m
Page 57 and 58: Table 2.3—Bone marker “seasonal
Page 59 and 60: Table 2.7—Chemistry panel “seas
Page 61 and 62: ABNormalized BSALPBSALP (U/L)CO N D
Page 63 and 64: Total Calcium (mg/dL)A1.21.151.11.0
Page 65 and 66: ABTotal OCN (ng/mL)150100500O N D J
Page 67 and 68: Adiponectin (ng/mL)AB40003500300025
Page 69 and 70: ABNormalized InsulinInsulin (μU/mL
Page 71 and 72: surrounding and encompassing hibern
Page 73 and 74: decreased from 788+30 ng/mL in preh
Page 75 and 76: 120-122]. Most small hibernators ar
Page 77 and 78: ears. Similarly, serum NPY increase
Page 79 and 80: Table 3.2—Serum factor longitudin
Page 81 and 82: Leptin (ng/mL)ABO N D J F M A M O N
Page 83 and 84: Norepinephrine (ng/mL)A807060504030
Page 85 and 86: IGF‐1 (ng/mL)AN D J F M A MB10009
Page 87 and 88: 2008. Samples were collected as des
Page 89 and 90: sampling points. It is possible to
Page 91 and 92: C‐Terminal PTH (Relative)AB504030
Page 93 and 94: Chapter Five — Serum from hiberna
Page 95 and 96: Synergy HT Multi-Detection Micropla
Page 97: gene specific primers and 12.5 μL
Page 101 and 102: hibernation cycles without problem
Page 103 and 104: filtered seasonal bear serum and fo
Page 105 and 106: which is dependent upon caspase-3 a
Page 107 and 108: Caspase‐3/7 ActivityO N D J F M A
Page 109 and 110: 6Caspase‐3/7 Activity54321**0Vehi
Page 111 and 112: ABCaspase‐3/7 Activity9876543210*
Page 113 and 114: eyond the G1/S checkpoint, thus ind
Page 115 and 116: ears, it is possible that tissue se
Page 117 and 118: PTH1R, RANKL, Runx2, Smurf1, TLR4,
Page 119 and 120: factors Runx2 and OCN also increase
Page 121 and 122: Table 6.3—Gene expression 3 hour
Page 123 and 124: Table 6.5—Gene expression 6 day d
Page 125 and 126: Cyclin D1 Gene ExpressionO N D J F
Page 127 and 128: Smurf1 Gene ExpressionO N D J F M A
Page 129 and 130: BAK Gene ExpressionO N D J F M A MM
Page 131 and 132: M‐CSF Gene ExpressionO N D J F M
Page 133 and 134: Per1 Gene ExpressionO N D J F M A M
Page 135 and 136: Akt Gene ExpressionO N D J F M A MM
Page 137 and 138: Bcl‐2 Gene ExpressionO N D J F M
Page 139 and 140: Cyclin D1 Gene ExpressionO N D J F
Page 141 and 142: OPN Gene ExpressionO N D J F M A MM
Page 143 and 144: 2PTH1R Gene Expression1.510.50O N D
Page 145 and 146: TLR4 Gene ExpressionO N D J F M A M
Page 147 and 148: Bcl‐2 Gene ExpressionO N D J F M
Page 149 and 150:
OCN Gene ExpressionO N D J F M A MM
Page 151 and 152:
PTH1R Gene ExpressionO N D J F M A
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TLR4 Gene ExpressionO N D J F M A M
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condition in renal patients in whic
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hibernation. This report brings to
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14. Kaneps, A.J., S.M. Stover, and
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41. Chowdhury, I., B. Tharakan, and
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68. Ashe, M.C., et al., Bone geomet
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95. Perrien, D.S., et al., Aging al
Page 167 and 168:
122. Lesser, R.W., et al., Renal re
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151. Confavreux, C.B., R.L. Levine,
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178. Toribara, T.Y., A.R. Terepka,
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206. Luo, X.H., et al., Adiponectin
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233. Florant, G.L., et al., Fat-cel
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259. Bhasin, S., et al., Older men
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284. Miura, M., et al., A crucial r
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Appendix A—Complete tables of cor
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Table A.5—Complete list of ionize
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Table A.7—Complete list of adipon
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Table A.12—Complete list of norep
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