Thesis (Ph. D.)--University of Washington, 2000. / Vita. Includes bibliographical references (leaves 53-58).

The loss of muscle mass leading to cachexia is rarely identified in early lung cancer. Fasting blood and muscle biopsy were collected in 59 non-small cell lung cancer (NSCLC) and 16 non-cancer patients, at the beginning of thoracic surgery. Serum C-reactive protein (CRP), and IL-6 were higher in NSCLC. In weight-losing NSCLC, food intake and serum albumin were lower, CRP, and TNF-alpha were higher. Although the expression of genes of the ubiquitin-proteasome system was not different, ubiquitinated-protein levels were lower and negatively correlated with ph-FOX01 in weight-losing patients. This would suggest lower muscle proteolytic rates in the early stages of NSCLC. Ph-FOXO1 also related to the degree of weight loss and stage of NSCLC. These data suggest that in early stages of the disease, weight and muscle loss could be mainly due to reduced food intake, rather than accelerated proteolysis, which reinforces the potential for successful dietary interventions to prevent or delay the onset of cachexia.

Muscle wasting is a significant complication of many diseases including diabetes mellitus, renal and liver failure, HIV/AIDS, and cancer. Sustained loss of skeletal muscle can severely impair a patient's quality of life and often results in poor tolerance and responsiveness to disease treatments. The increased protein breakdown observed during muscle atrophy has been attributed to accelerated activity of the ubiquitin-proteasome pathway, but the precise mechanisms by which this activation stimulates muscle protein loss are poorly understood. Previous work showed that the deubiquitinating enzyme USP19 is upregulated in rat skeletal muscle in various forms of muscle wasting, including streptozotocin induced diabetes, cancer, and dexamethasone treatment. 1 To further explore the role of USP19 in muscle wasting, siRNA-mediated depletion of the enzyme was carried out in L6 myotubes. Knockdown of USP19 resulted in more rapid differentiation of myoblasts into myotubes, with a greater extent of myoblast fusion. It also produced tubes that were visibly larger than those formed by myoblasts transfected with a control siRNA. At the molecular level, silencing of USP19 increased the amount of myosin heavy chain (MHC) and tropomyosin proteins. It also increased levels of MHC transcript, suggesting that USP19 acts at the level of gene transcription or mRNA stability rather than protein degradation. USP19 may mediate its effects on muscle-specific gene expression through the myogenic transcription factor myogenin, since depletion of USP19 increased protein and mRNA levels myogenin but did not affect protein levels of the related transcription factor Myf5. Moreover, the increased tropomyosin and MHC observed upon USP19 knockdown could be abolished when myogenin was simultaneously depleted using siRNA. Collectively, these results suggest that USP19 functions to inhibit the synthesis of key muscle proteins and may therefore be a promising target for the treatment of muscle atrophy.

Progressive resistance training (PRT) in the elderly has commonly used ATPase histochemistry to evaluate fiber type changes, but evidence shows there are myosin heavy chain (MHC) hybrids in aging muscle that cannot be classified by histochemistry. The purpose of this study was to assess the MHC and whole muscle alterations following a 12-week PRT protocol. Seven healthy men (age=74.0±4.7, weight=74.6±13.5kg) underwent testing for 1-repetition maximum (1-RM), whole muscle (thigh) crosssectional area (CSA) by computed tomography, and a needle muscle biopsy from the vastus lateralis for analysis of MHC, pre- and post-training. The PRT consisted of 2 sets of 10 repetitions, and a third set to volitional exhaustion at 80% 1-RM, 3 days per week for 12 weeks. Muscle ATPase histochemistry analysis for distribution did not significantly differ following training. Muscle samples were freeze dried and dissected for MHC analysis (sodium dodecyl sulfate-polyacrylamide gel electrophoresis (5% gel) and silver stained; 224.0±11.2 and 213.0±8.1 fibers/subject pre-/post-training; total fibers analyzed=3059). MHC analysis demonstrated significant increases in MHC I proportion (10.4%; P<0.05), and significant decreases in MHC UIIa (9.0%; P<0.05), UIIa/x (0.9%; P<0.05), and IIa/x (8.9%; P<0.05) isofroms, along with no change in the MHC Ila and IIx isoforms, pre- versus post-training. In addition, 1-RM (51.9%; P<0.05) and CSA (5.9%; P<0.05) increased from pre- to post-testing. This data supports previous whole muscle changes, more important, is the increase in MHC I and decrease in MHC I/IIa, I/IIa/IIx, and IIa/x hybrids. The myosin light chain 3f (MLC3f) to MLC 2 ratio did not change with the PRT in either the MHC I or MHC IIa isoforms, although there was a significantly greater amount of MLC 3f in the MHC Ila versus the MHC I fibers (p<0.05), pre- and post-training. The myosin isofrom data provides support that aging muscle has the plasticity to adapt in a manner unlike that of young muscle. / School of Physical Education

Access to abstract permanently restricted to Ball State community only / Access to thesis permanently restricted to Ball State community only / School of Physical Education, Sport, and Exercise Science

We have previously shown that non-specific blockade of the cyclooxygenase (COX) enzymes in skeletal muscle eliminates the normal increase in muscle protein synthesis following resistance exercise. The current study tested the hypothesis that this COX-mediated increase in postexercise muscle protein synthesis is specifically regulated by the COX-2 isoform. Sixteen males (23 ± 1 yr, 177 ± 2 cm, 81.5 ± 3.4 kg) were randomly assigned to one of two groups that received three doses of either a specific COX-2 inhibitor (celecoxib; 200 mg per dose, 600 mg total) or a placebo during the 24 hours following a single bout of resistance exercise with the knee extensors. Skeletal muscle fractional synthesis rate (FSR) was measured at rest and 24 hours postexercise using a primed constant infusion of [2H5]phenylalanine coupled with muscle biopsies of the vastus lateralis. Mixed muscle FSR was increased following exercise to a greater extent (206%, P<0.05) in the COX-2 group (0.052 ± 0.014 %Ih) as compared with the placebo group (0.017 ± 0.007 %Ih). These results suggest that the specific inhibition of the COX-2 isoform in human skeletal muscle causes a compensatory response in muscle protein synthesis. These data also highlight the involvement of the cyclooxygenase pathways in the regulation of muscle protein synthesis following resistance exercise. / School of Physical Education, Sport, and Exercise Science

I investigated SL trans-splicing in the troponin I gene of Ciona intestinalis. Experimental mutation of the AG dinucleotide adjacent to the natural trans-splice acceptor site (-64) in CiTnI/nuclacZ constructs eliminated trans-splicing to that site in Ciona embryos but activated trans-splicing at cryptic acceptor sites at -76 and -39, adjacent to the nearest AG dinucleotides. However, not all AG dinucleotides specify cryptic acceptor sites because outron internal deletions or 3'truncation mutants were trans-spliced at a far-upstream AG-adjacent cryptic site (-346), leaving many AGs in the retained outron segments. Thus, additional sequence elements that are present only in the -346 and -76/-64/-39 regions are required for cryptic acceptor activity. All mutant constructs generated detectable beta-gal enzyme expression, although the mutant with the longest retained-outron segment appeared less active. Therefore, mRNA accumulation and translation do not require trans-splicing to the natural acceptor site, although they may be facilitated by the normal removal of the outron during trans-splicing.

Thesis (Ph.D.)--Baylor University, 2008. / Includes bibliographical references (p. 121-133)

Thesis (Ph. D.)--University of Missouri--Columbia, 2002. / "May 2002." Typescript. Vita. Includes bibliographical references (leaves 103-110).

Thesis (M.Sc. )--Memorial University of Newfoundland, 1997. / Bibliography: leaves 137-145.