Department of Medicine Presentations (UMKC)
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Items in this collection are the scholarly output of the Department of Medicine faculty, staff, and students, either alone or as co-authors, and which may or may not have been published in an alternate format
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Item Hyperthermia Induces Functional and Molecular Modifications in Cardiac, Smooth and Skeletal Muscle Cells(2010-03) Romero, Sandra; Hall, Todd; Touchberry, Chad; Elmore, Chris; Silswal, Neerupma; Parelkar, Nickil; Baker, Kendra; Loghry, Michael; Rizk, Hatem Ibrahim; Mo, Chenglin; Brotto, Leticia; Leon-Salas, Walter D. (Walter Daniel); Wacker, Michael J.; Andresen, Jon; Brotto, Marco; University of Missouri (System); Missouri Life Sciences Summit (2010: University of Missouri--Kansas City)Hyperthermia is used for the treatment of a number of diseases, including muscle injuries, inflammations, tendinitis, and osteoarticular disorder. More recently, hyperthermia has been used as an adjuvant in cancer treatment. Only two studies have shown that hyperthermia leads to hypertrophy in in-vitro models of cardiac and skeletal muscle cells. Functional, biochemical and molecular mechanisms of hyperthermia-induced hypertrophy in muscles remain largely undiscovered. We investigated the effects of mild heat shock (HS) on C2C12 skeletal, HL-1 cardiac and AR-75 smooth muscle cells. Mild HS (20 min 43ºC) induced increases in the cell area in all muscle cells tested. C2C12 cells are a well-accepted model of skeletal muscle fibers, and were selected for complementary studies. First, to biochemically confirm an increase in protein synthesis we measured and found an increase of ~6% in total protein content 24 hrs after HS. Second, we examined potential modifications in calcium (Ca) homeostasis regulation by measuring intracellular Ca. We detected a lower resting level of intracellular Ca and smaller and longer caffeine-induced Ca transients in C2C12 muscle cells 24 hrs after HS. Next, to search for molecular mechanisms involved with HS-induced hypertrophy and calcium homeostasis modifications, mRNA from C2C12 muscle cells was analyzed at different time points after HS (0, 1, 2, and 24 hrs). We used an ABI Step One Plus RT2 PCR Array System and a custom-built 96 gene array. We report for the first time that the expression of key heat-shock, hypertrophy/ metabolic, and Ca+2 signaling genes were altered after HS. Hsp70 and Hsp72 genes were highly expressed (211-1829 fold change) after HS. Also, Myh7 (MHC-I), Myh6, Srf, Ppp3r1 and Pck1 were up-regulated by 2-6 fold change compared with control cells.. Furthermore, a reduction in the expression of RyR and Trdn genes was observed (2- 3.6 fold change) with an associated increase in the expression of IP3R genes (2-4 fold change). These results indicate that hyperthermia modulates not only heat-shock related and hypertrophy genes, but also genes involved with metabolism, apoptosis repression, calcium homeostasis and signaling, and cell homeostasis. Our studies offer an initial exploration of the functional, biochemical and molecular mechanisms that may help explain the beneficially adaptive effects of hyperthermia on muscle function. Our studies shall also prove useful for the refinement of a specific device (EM-Stim) to be employed for the treatment of muscle and bone diseases (See poster by Hatem et al). Importantly, our studies have potential translational applications. By learning how to more precisely use hyperthermia to control specific genes that can improve or treat muscle injuries, musculoskeletal, and cardiovascular diseases, the ensuing benefits shall be unmistakable. Our short and long-term goals are: i) optimize our protocols; ii) test HS in animal models; iii) manipulate expression of promising genes of interest in vitro and in in-vivo animal models; iv) initiate clinical studies to fully translate from the bench to the bed-side.Item Clearing the hurdles for nanotechnology: In vivo inhalation effects(2010-03) Herndon, B.; Nalvarte-Kostoryz, Elisabet L. (Elisabet Lourdes), 1958-; Molteni, A.; Quinn, T.; Fibuch, Eugene; University of Missouri (System); Missouri Life Sciences Summit (2010: University of Missouri--Kansas City)INTRODUCTION: Nanoparticles of many types have been created for industrial and medical applications. Among these nanoparticles, single-walled carbon nanotubes (SWCNT) are of high interest for their physicochemical properties and application in electronics, drug delivery and other areas. With the rapid expansion in SWCNT-based new technologies, a full understanding of their safety and risks for human exposure must be considered. Because of the potential human risk of nanoparticle exposure we have developed an animal model to study the effects of nanoparticle exposure on lung tissue. Using this rat model we hypothesized that an acute nanoparticle exposure would result in an inflammatory response in lung tissue. METHODS: Particle instillation (intratracheal under direct visualization) of 50 μL pediatric surfactant containing 500 micrograms SWCNT (or surfactant alone) was performed in 32 rats to date. Pulmonary histology and biochemical measures on bronchoalveolar lavage (BAL), pleural fluid, serum and lung cells was quantified. RESULTS: Very early (<30 minutes) eosinophilia developed in lung tissue following SWCNT instillation. Innate immune system sterile response, or Damage Associated Molecular Patterns (DAMPs) protein was released. Our dose proved sterile, <0.03 EU LPS, showing the effect was damage-induced not pathogen-induced. High mobility group box protein-1 (HMGB1), a nuclear chaperone and prototype DAMP was elevated (ELISA) following SWCNT exposure. A second DAMP, heatshock protein 70 (HSP-70), a cytoplasmic chaperone, was also quantified by ELISA. The response OF HSP-70 over time is similar to HMGB1. Western blots performed on time-harvested lungs exposed to SWCNT demonstrated a receptor for advanced glycation end products (RAGE), with a strong peak at 3 hours after pulmonary exposure. The inflammatory cytokine TNFα appeared in lung tissue and bronchial alveolar lavage (BAL) at 30 minutes, with the same timing as the HMGB1 and HSP-70 release. Flow cytometry of type II pneumocytes and pulmonary macrophages from SWCNT-exposed rats demonstrated secondary DAMP receptors. A potential chronic effect was noted at one month. HMGB1 and HSP-70 peaked acutely at approximately 24 hr and then slowly decreased at 1 to 2 weeks. At 1 month, however, a new increase was seen. CONCLUSIONS: The hydrophobic SWCNT, important industrial components, form bundles and fibers in the hydrophilic lung, creating an immediate cellular inflammatory response, measurable cellular necrosis and very rapid chemokine release. Early data suggests the potential for chronicity.