Browsing by Subject "Burns"
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Item Burn Serum Stimulated Mitochondrial Fission Was Decreased with IL-6 Antibody Treatment(2017-01-17) Sehat, Alvand; Song, Juquan; Kulangara, Rohan; Maredia, Navin; Maxwell, Christian; Panwar, Kunal; Huebinger, Ryan M.; Carlson, Deborah L.; Zang, Qun S.; Wolf, Steven E.INTRODUCTION: Burn patients suffer muscle mass loss associated with hyperinflammation and hypercatabolism. We previously observed that burn serum resulted in cell death with elevated mitochondrial fragmentation in C2C12 myoblast. IL-6 as the key cytokine response to thermal injury has been showed to increase mitochondrial fragmentation. We thus posit that inhibition of IL-6 expression in burn serum will alleviate mitochondrial fragmentation. The aim of this study is to investigate the neutralization effect of IL-6 antibody in burn serum stimulated myoblasts. METHODS: Murine myoblasts C2C12 cells were cultured with recombinant IL-6 protein from 0.01 ng/ml, 0.1 ng/ml, 1 ng/ml, and 100 ng/ml. Cells were labeled with MitoTracker Green dye and live cell images were captured with Confocal microscopy. Next, C2C12 cells were exposed to medium containing 1) 10% serum from control rat, 2) 10% serum from burn rat, and 3) 10% serum from burn rat and 0.5 ug/ml of IL-6 antibody. All cells were labeled as the first experiment and live cell images were recorded. The caspase 3 activity was further examined from cell protein lysate. RESULTS: The 1 ng/ml dose of r-IL6 showed a fourfold increase in mitochondrial volume (μm3) at 24 hours post challenge. The intensity signal of Mitotracker was significantly increased in the 1 ng/ml IL-6 dose group at 48 hours. The 1 ng/ml dose of r-IL6 showed an increase in mitochondrial fragmentation. In cells cultured with 10% serum from control rats, mitochondrial morphology maintained the elongated linear shape during the 48 hours. In cells cultured with 10% serum from burned rats, a reduction in mitochondrial sizes was significant. In cells cultured with 10% serum and IL-6 antibody treatment this effect was reversed. Further, the intensity signals increased in response to the burn serum challenge. However, with addition of IL-6 antibody treatment the intensity signals decreased. In addition, burn serum increased the total volume of mitochondria about 1.4 fold, while with IL-6 antibody treatment the total volume was decreased. Consistently, a significant decrease in the expression of caspase 3 in the IL-6 antibody treatment group was observed. CONCLUSION: IL-6 stimulates an increase in mitochondria fragmentation in myoblasts, while IL-6 antibody treatment decreases mitochondrial fragmentation and cell death in burn serum stimulated myoblasts. This project has shown that targeting cytokine levels may be an effective treatment strategy in the management of burn patients.Item Burn Serum Stimulates Mitochondrial Fission in C2C12 Myoblasts(2021-04-22) Sehat, Alvand; Wolf, Steven; Carlson, Deborah; Huebinger, RyanBACKGROUND: Burn patients suffer muscle mass loss associated with a hypercatabolic status. Impairment of mitochondrial function has been observed in the muscle of burn patient's. OBJECTIVE: We hypothesize that muscle atrophy due to burn injur y is associated with an alteration in mitochondrial dynamics. This study was designed to investigate changes in mitochondrial fission and fusion in response to a burn serum challenge in myoblasts in vitro. METHODS: Cultured murine myoblasts, C2C12 cells, w ere exposed to 10% rat serum isolated either from 40% total body surface area (TBSA) scald burn rats or control rats. Cells were then labeled with MitoTracker Green dye and live cell images were recorded by confocal microscopy. The expression of mitochondr ial fission/fusion factors was examined by Western blots. RESULTS: In cells cultured with 10% serum from control rats, mitochondrial morphology maintained the elongated linear shape during the 48 hours we observed. However, 24 hours of culturing in 10% scaled rat serum resulted in a significant reduction in mitochondrial size. .Further, the number of total mitochondria increased, indicating a stimulation of mitochondrial fragmentation in response to the burn serum challenge. In addition, burn serum increases the total volume of mitochondria about 1.4 fold. Consistently, Western blot analysis showed a significant decrease in the expression of mitochondrial fusion protein Mfn1. CONCLUSION: Burn serum stimulates an increase in mitochondria fission in myoblasts.Item Epidemiology of Burns Treated by Yekatit 12 Burn Unit in 2016(2017-04-03) Mehta, Kajal Anil; Abdelnaby, Abier; Chang, Mary; Phelan, HerbBACKGROUND: Burns contribute significantly to the burden of disease, particularly in low and middle-income countries. There is a paucity of data regarding the epidemiology of burns in Ethiopia, and about burn injuries in Ethiopian urban settings. This study describes the epidemiology of burn injury admissions in a public tertiary care hospital burn unit. METHODS: A retrospective observational study was performed and the hospital records of the burn unit admissions, operating room logs and collected data sheets were utilized. All patients treated on an inpatient basis by the Yekatit 12 Hospital Burn Unit team between December 2015 and December 2016 were included in the study. RESULTS: A total of 184 burn hospitalizations were registered during this time period. Of the registered patients, 42.4% were female and 57.1% were male. The average age of the patients was 16.4 year old; 51.1% of patients were pediatric (<14 years old) with the average pediatric patient age of 4.36 years and average adult patient age of 29.4 years. Flame burns accounted for 35.8% of burns, scald 35.1%, electrical 27.8% and others <0.01%. The gender ratio was significantly dependent on the cause of burn (X2(10, N=184) = 35.9, p<0.0001), with males more likely to sustain electrical and scald burns (10.25 and 1.19 times the rate seen in females). The average total body surface area (TBSA) burned as calculated on the Lund-Browder scale, was 19.8% (n=77, SD=18.2). In admitted patients, 52.7% underwent operative treatment, with most frequent operation being split-thickness skin graft (STSG). The mortality rate was 13.3% (n=105). CONCLUSION: This is the most recent epidemiological study of burn patients in an urban setting in the last 15 years in Ethiopia. This research highlights the current picture of burn injuries in Addis Ababa. Many of the results are consistent with the literature of scald and flame burns being the predominant etiology, and a mortality rate of 13.3%. Though this study is limited by incomplete data, the proportion of burns due to electrical burns was higher than has been previously demonstrated, and thus warrants further study.Item Functional Outcomes in Patients with Full Thickness Hand Burns(2020-03-10) Vu, Kevin Quochuy; Holavanahalli, Radha; Kowalske, Karen; Hynan, Linda S.BACKGROUND: There has been previous work objectively examining the severe contractures that develop with hand burns, but few have correlated severity with functional outcome. While contracture definitions exist for restrictions in range of motion (ROM), they have not been linked to prognostic use and functional outcome. OBJECTIVE: The objective of this study is to correlate severity of hand contracture in joints of the hand with differences in functional outcomes scores at discharge. METHODS: This multicenter study uses the National Institute on Disability, Independent Living, and Rehabilitation Research (NIDILRR) database, otherwise known as the Burn Injury Model Systems (BMS) National Database. A sample of 95 patients with ROM measurements and the SF-36 physical component score (PCS) to measure functional outcome was used. Patients were grouped by ROM into mild, moderate, or severe contracture definitions. Inclusion criteria included those with burn injuries as defined by the American Burn Association who had recorded ROM deficits. Patients with post-injury amputations were excluded. Statistical analyses were conducted to compare the maximum severity of contracture in both hands on the PCS when the maximum contracture was classified as mild, moderate, or severe. Secondary analysis was used to also compare PCS between mild and moderate versus severe contracture groups. RESULTS: There was no significant difference in PCS for mild, moderate or severe contracture (p = 0.858). There was a downward trend noted in the means between each contracture group, as well as several outliers in the moderate contracture group. Secondary analysis between a combined mild and moderate versus severe contracture group showed no significant difference in PCS between the two groups (p = 0.654) CONCLUSION: The results of this study suggest that although there is a downward trend in PCS that correlates with severity of contracture, the difference in functional outcome as measured by the PCS is not significant between the different ratings of contracture severity. Future studies involving long-term PCSItem Functional, Psychological and Community Integration Changes Over Time in Persons With Major Burn Injury(2006-05-15) Grace, Sheila Ann Isom; Cromes, G. FredThe purpose of the present study was to measure prospectively the functional, psychological and community integration status of individuals with major burn injury in order to determine the extent of changes over time and how functional and community integration status relates to emotional distress. This study was part of the North Texas Burn Model System Grant funded by the National Institute on Disability and Rehabilitation Research. Subjects in this study included adults over 14 years of age who met the American Burn Association's criteria for a major burn injury. The Total Body Surface Area burn across the measurement periods in these subjects ranged from 22.4 to 25.2 percent. Data was collected from 356 participants at discharge, 199 participants at 2 months, 255 participants at 6 months, 193 participants at 12 months, 114 participants at 24 months and 41 participants at 36 months. The instruments used in this study were the Brief Symptom Inventory (BSI), the Burn Specific Health Scale (BSHS), The Community Integration Questionnaire (CIQ), the Pain Analog Scale (PAS) and the Functional Assessment Screening Questionnaire (FASQ). It was hypothesized that 1) both areas of functioning and community integration would improve as emotional distress abated and that 2) the functional improvement and community integration as well as the lessening of emotional distress would continue over time. A repeated measures ANOVA was used to measure changes over time and a Bonferroni Multiple Comparison Test was used to compare results at different time intervals. FASQ and CIQ total scores for individuals exhibiting high versus low measures of emotional distress (with median split of BSI global scores) were compared at each measurement period using the Student's T-test. A statistically significant interaction was found between the variables as predicted; however, the participants in this study appeared to reach a plateau at 24 months after which little change in outcomes was noted.Item Gene Expression Changes in Response to Severe Burn Injury in Rat Achilles Tendon(2017-01-17) Buller, Dustin; Mitchell, Wes; Manchanda, Kshitij; Song, Juquan; Hernandez, PaulaIMPORTANCE: Severe burn injury, occurring at a global rate of 5/100,000 per year, can result in serious sequelae such as infection, shock, and direct organ damage. It has been shown that severe burn also results in both systemic inflammation and distant inflammatory effects, specifically in skeletal muscle and bone. However, we are unaware of any work that investigates similar effects in the tendon, which connects muscle and bone. OBJECTIVE: To determine whether, after distant severe burn injury, changes occur in gene expression, protein synthesis, and biomechanical properties in rat Achilles tendon. METHODS: Rats were subjected to thoracolumbar full-thickness severe burn injury under anesthesia before harvesting the Achilles tendon at time points of 6h, 1d, 3d, 7d, and 14d. Unburned rats were used for control. RNA expression of collagen I (Col1), collagen III (Col3), MMP9, MMP13, IL-1β, IL-6, TNF-α, and tenomodulin was measured using qPCR. Quantified Western blots were performed to assess relative protein content for Col1, Col3, and MMP9. Finally, biomechanical testing was performed on a separate group of rats to assess biomechanical changes at 14d versus control (n = 8). RESULTS: The MMP9 gene was upregulated on the order of 20-fold at 14d (p = 0.0016) and 10-fold at 3d (p = 0.0255) and MMP13 showed a trend toward 12-fold increase at 3d (p = 0.079). A significant increase in expression of IL-1β and a trend toward significance IL-6 were also observed at 3d (p < 0.01, and p = 0.059, respectively). Differences in expression of Col1, Col3, TNF-α, and tenomodulin were not significant compared to control. Preliminary data from Western blots showed 5-fold decrease of collagen 1 at 7d (p < 0.01) and 2-fold increase of collagen 3 at 14d (p = 0.016). Significant changes were not found in MMP9. Preliminary biomechanical data shows a trend toward a 4-fold decrease in stiffness in the burn group. All p values are by t-test. CONCLUSIONS: These results newly confirm the existence of inflammation in tendon distant from the burn site after severe burn injury. Specifically, these changes could indicate initial matrix remodeling as carried out by the MMPs beginning at 3d, followed by collagen deposition with a decreased Col1:Col3 ratio, resulting in decreased tissue stiffness. These findings are consistent with the repair processes known to occur in other tissues after inflammation. Additionally, IL-6 and IL-1β may have a more significant role in post-burn acute-phase inflammation than other acute-phase reactants like TNF-α in the Achilles tendon.Item Inflammatory Effects of Severe Burn Injury in Rat Intervertebral Disc(2017-01-17) Mitchell, Thomas Wes; Buller, Dustin; Manchanda, Kshitij; Song, Juquan; Hernandez, PaulaIMPORTANCE: Severe burn injury leads to a myriad of clinical effects, some of which are mediated by systemic inflammatory cytokines. Although inflammation has been shown to increase skeletal muscle catabolism, little is known about the effects this inflammation has on the intervertebral disc tissue and whether or not it predisposes to spinal pathology. OBJECTIVE: To investigate the gene expression changes after severe burn injury in rat intervertebral disc tissue and explore the possible role of inflammatory cytokines in tissue remodeling imbalance. METHODS: Rats were given severe burn injury according to protocol, then harvested at multiple time points: 6h, 1d, 3d, 7d, and 14d post-burn. Unburned rats used for control. Rat intervertebral discs were dissected and removed, then used for measurement of gene expression using qPCR. RESULTS: Differences in gene expression of structural matrix proteins between time points were not statistically significant, however, TNF-alpha showed statistically significant (p<.05) decreased expression at 1d, after increased expression at 6h that approached significance (p=.055); this pattern of gene expression is consistent with previous results. In addition, there was a statistically significant (p<.05) increase in expression of the TRPV4 channel, which helps regulates osmolarity of the tissue in the nucleus pulposis. This could explain how osmotic dysregulation induced by pro-inflammatory cytokines leads to loss of water content from nucleus pulposis cells, thus triggering disc degeneration. CONCLUSIONS: These results indicate that in the post-burn state pro-inflammatory cytokines, such as TNF-alpha, likely have an effect on intervertebral disc tissue and the subsequent up-regulation of TRPV4 could have long-term effects that hasten the onset of disc degeneration related to loss of water content.Item Mitochondrial Dynamics in Renal Cell with Burn Serum Stimulation: An Observational Study(2017-01-17) Maxwell, Christian T. W.; Huebinger, Ryan M.; Song, Juquan; Sehat, Alvand J.; Wolf, Steven E.BACKGROUND: Myoblasts and myocardiocytes have increased cell death with burn serum stimulation, which is associated with mitochondrial fission and function impairment. Acute kidney injury is a significant issue in burn patients; however, the role of the mitochondrial response in renal cells (RCs) has not been explored. The purpose of this summer project was to establish a method that observes mitochondrial dynamics in renal epithelial cells upon exposure to burn serum. HYPOTHESIS: The mitochondrial fission/fusion cycle in human RCs is disrupted following exposure to burn serum. METHODS: Human primary renal proximal tubule epithelial cells were cultured with Renal Epithelial Cell Growth Media in an incubator at 37°C with 5% CO2. Upon reaching 65-70% confluency, RCs were treated with growth media containing 10% rat serum (RS) either from control rats or 40% total body surface area scald burn rats. 3nM of MitoTracker Green FM dye was added to all cell treatments to stain the mitochondria (MT). Live cell images were taken under a Nikon Ti Eclipse Confocal microscope at 6, 24, 48, 72, and 96 hour time points; subsequently, images were analyzed with Nikon software to identify fluorescent intensity, mitochondrial volume and elongation. T-tests were then applied to analyze the significance with Bonferroni correction. Significance after correction was determined if p < .01 . OUTCOMES: RC morphology was cuboidal and refractile in the culture media. During serum stimulation experiments, the MT of RCs gradually increased in fluorescent intensity and maintained a rod-like shape in both groups. Specifically, the MT exposed to normal RS had significantly higher intensity at the 72 and 96 hour marks than those in burn RS. Despite these results, cell viability was determined to be 48 hours, since 60% or more of cells had detached at the 72 and 96 hour marks, indicating cell death. There were no significant differences in elongation and volume between burn and normal serum stimulation at each time point. CONCLUSIONS: The cell culture protocol for human primary renal epithelial cells was established this summer. Although rats and humans share evolutionarily conserved traits, the RS did not significantly influence the more evolved human RC.Item Mitochondrial Fission with Function Impairment in Burn Serum Treated C2C12 Cells(2016-01-19) Sehat, Alvand; Song, Juquan; Kumar, Puneet; Cai, Anthony; Huebinger, Ryan M.; Carlson, Deborah, L.; Zang, Qun S.; Wolf, Steven E.BACKGROUND: Burn patients suffer muscle mass loss associated with a hypercatabolic status. Mitochondria dynamics cycle is affected by metabolic status, and mitochondrial fission mediated high glucose induced cell death. Mitochondria function impairment associated with muscle mass loss has been observed in severe burn patients. We hypothesize that severe burn impaired muscle mass loss is associated with increased mitochondria fission with function impairment. The study was to investigate mitochondrial dynamics in response to burn serum stimulation. METHODS: Murine myoblast C2C12 cells were treated with DMEM media containing 10% rat serum isolated either from 40% TBSA scald burn rats, or control rats. Mitochondria was labeled with 3nM of MitoTracker Green FM dye, and live cell images were taken sequentially under a Nikon Ti Eclipse Confocal microscope. Cell lysates were collected for molecular biological analysis. Mitochondrial function was evaluated with Enzo Mito-ID membrane potential cytotoxicity kit. Target protein signals from cell lysate were detected by SDS-PAGE and western blot analysis. RESULTS: Mitochondrial morphology maintained the elongated linear shape in C2C12 cells when treated with 10% control rat serum. In contrast, when cells were treated with 10% burn serum, mitochondria reduced the elongated linear shape at 24 to 48 hours, and the florescent dye diffused at 72 hours. The cell florescent images showed an increase in circularity and fragmentation of mitochondria in C2C12 cells with burn serum stimulation. Meanwhile mitochondrial membrane potential decreased with 6hr post-burn serum stimulation. Western blot data showed that mitofusion-1 (Mfn1) significantly decreased in C2C12 cells with burn serum stimulation, confirming the observation of mitochondrial fission in response to burn serum. Cell death marker caspase 3 increased its expression in C2C12 cells with burn serum stimulation, suggesting a superfluous cell death in skeletal muscle after burn. CONCLUSION: Our results show an increase in the mitochondria fission/fusion ratio in C2C12 cells stimulated with burn serum isolated 6 hours after burn. The mechanism of mitochondrial fission with function impairment leading to muscle death is under investigation.Item Myokine Musclin Expression Is Elevated in Rats after Burn(2017-01-17) Maredia, Navin; Song, Juquan; Sehat, Alvand; Maxwell, Christian; Panwar, Kunal; Kulangara, Rohan; Carlson, Deborah; Huebinger, Ryan; Wolf, StevenINTRODUCTION: Annually, over 2 million people in the US experience severe burns, a condition marked by a hypercatabolic state with significant muscle loss. Muscle is necessary for glucose and lipid metabolism. Previous studies have shown detrimental effects of insulin resistance and hyperglycemia associated with muscle loss due to burns. Recently, the novel skeletal muscle myokine musclin has been found to regulate glucose in vitro. Thus, we attempted to better understand the effects of burns on musclin levels. We aimed to investigate the effects of burns on 1.) musclin levels systemically and 2.) musclin mRNA expression in vitro. METHODS: Thirty-one adult male Sprague-Dawley rats received 40% total body surface area (TBSA) burns. Rat serum was collected from 6 hours to 14 days after burn. Nine animals without injury served as control. Musclin levels in serum were measured by ELISA. Mouse C2C12 myoblasts were stimulated with 10% rat burn serum for 24 hours. Cells were stimulated with non-burn serum, 6-hour post burn serum, 72-hour post burn serum, and 14-day post burn serum. Following stimulation for 24 hours, C2C12 cells were collected and musclin expression was quantified by real time PCR analysis. RESULTS: Circulating musclin levels were 59.3 ± 3.3 ng/mL in non burned control rats. Musclin levels in the serum significantly increased to 76.7 ± 6.0 ng/mL at 6 hours and 76.7 ± 1.9 ng/mL at 24 hours after burn (p<0.05). Musclin levels in the serum returned to baseline until 14 days. Normalized to GAPDH mRNA level, musclin mRNA expression was 7.87 ± 0.56 fold in C2C12 myoblasts with 10% non-burn serum stimulation. Musclin mRNA expression significantly increased with the addition of the following burn rat serums: 22.66 ± 5.18 fold with 6-hour post-burn serum, 15.00 ± 1.93 fold with 72-hour post-burn serum, and 12.17 ± 0.82 fold with 14-day post-burn serum (p<0.05). CONCLUSIONS: Musclin levels increase in rat serum following 40% TBSA burn injury. In vitro stimulation of muscle cells with burn serum increases musclin expression.Item [News Items](1953-05-15) Aagaard, George N.Item [News](1977-04-14) Land, ChrisItem [News](1980) Harrell, AnnItem [News](1980-10-13) Harrell, AnnItem [News](1973-04-02) Fenley, Bob; Harrell, AnnItem [News](1974-11-07) Taylor, SilviItem [News](1982-01-18) Harrell, AnnItem [News](1981-08-07) Harrell, AnnItem [News](1984-08-31) Harrell, AnnItem [News](1973-04-05) Fenley, Bob
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