669 Application of In-cell Western Blot to Study Burn-induced Cardiac Dysfunction

2021 ◽  
Vol 42 (Supplement_1) ◽  
pp. S191-S191
Author(s):  
Jake J Wen
Keyword(s):  
Mitochondrial Biogenesis ◽  
Cardiac Dysfunction ◽  
Cultured Cells ◽  
Burn Injury ◽  
Total Body Surface Area ◽  
Enzymic Activity ◽  
Good Tool ◽  
Human Cardiomyocytes ◽  
Related Proteins ◽  
In Cell Western

Abstract Introduction Traditional Western blotting (WB) may not reflect true target protein amount due to interference from enzymic activity during sample preparation. In-Cell Western (ICW) blotting has proved to have advantages over the traditional WB. However, ICW has not been used to study burn-induced cardiac dysfunction. This study will explore the efficacy of ICW in the study of burn-induced cardiac dysfunction. Methods Human cardiomyocytes (Ac16) were cultured in serum from rats with/without 60% of total body surface area (TBSA) burn injury. Cultured cells were permeabilized by Tritons-100 X, 1st anti-targeted protein antibodies were added and incubated, and, finally, two fluorescence-conjugated 2nd antibodies (one red and one blue) were added and incubated. LI-COR Odyssey CLx scanner was employed to obtain images and LI-COR Image Studio 4.0 software was used to analyze image and GraphPad Prism 8.0 software was used to determine image density. Results ICW proved to be a good tool to study burn injury. Our ICW data showed that burn serum resulted in 1) increased cardiomyocyte PARP1 (leading to increased protein acetylation); 2) increased damage of cardiomyocyte mitochondrial DNA replication (via increase in mitochondrial Fis1and BCBN proteins and decrease in parkin and POLG proteins); and 3) a decrease of cardiomyocyte mitochondrial biogenesis (with increase in inflammation-related proteins, apoptosis-related proteins, and decrease Brf2-ARE-related proteins). Conclusions This study provides preliminary evidence that ICW may be a novel tool to study burn injury and confirms that burn-injury induced cardiomyocyte mitochondrial dysfunction occurs via damage of mitDNA replication, involving inflammation, apoptosis, and mitochondrial biogenesis pathways.

Is the Use of Routine Echocardiogram Warranted in Large Pediatric Burns?

2021 ◽  
Author(s):  
I Wilmot ◽  
P H Chang ◽  
L Fowler ◽  
P Warner
Keyword(s):  
Left Ventricular Dysfunction ◽  
Cardiac Dysfunction ◽  
Ventricular Dysfunction ◽  
Burn Injury ◽  
Early Recognition ◽  
Total Body Surface Area ◽  
Left Ventricular ◽  
Burn Patients ◽  
Severe Left Ventricular Dysfunction ◽  
Pediatric Burns

Abstract Introduction Cardiac dysfunction can develop in large pediatric burns during the acute and recovery phase. When occurring in this population, the cardiac abnormality appears as left ventricular dysfunction or dilated cardiomyopathy. Recent studies have demonstrated perioperative and long-term cardiac dysfunction resulting in longer hospital stays for patients over 40% total body surface area. The objective of this study was to assess if early use of echocardiograms in large burns would allow for early recognition of patients at risk for cardiac dysfunction. Material and Methods Pediatric burn patients ages 0-18 years who sustained a burn injury of 30% TBSA or more or developed cardiac dysfunction during hospital course were evaluated. Echocardiograms were obtained upon admission with monthly repeats until three normal studies were attained or the patient was discharged and when symptomatic. Results Of the 130 acute burn patients admitted during 7/2017-10/2018, 10 patients met criteria for enrollment in this study. The average age was 5 years (0.8 to 10 yrs), 70% were males and 90% sustained flame injuries.Total TBSA average was 45% (24-70%) with average full thickness burns of 33% (0-67%). Twenty echocardiogram studies were obtained. One patient with 25% TBSA burn, demonstrated severe left ventricular dysfunction with an EF of 25% from post arrest myocardial stunning. Repeat echocardiogram studies demonstrated full recovery with normal EF. The remaining patients, despite large TBSA injuries, did not exhibit any abnormalities on ECHO examinations. No cardiac interventions were required. Conclusions Use of echocardiograms is best performed on symptomatic burn patient populations.

Macrophage migration inhibitory factor mediates late cardiac dysfunction after burn injury

2005 ◽  
Vol 288 (2) ◽  
pp. H795-H804 ◽  
Author(s):  
Monte S. Willis ◽  
Deborah L. Carlson ◽  
J. Michael DiMaio ◽  
Michael D. White ◽  
D. Jean White ◽  
...  
Keyword(s):  
Cardiac Function ◽  
Migration Inhibitory Factor ◽  
Cardiac Dysfunction ◽  
Macrophage Migration Inhibitory Factor ◽  
Burn Injury ◽  
Total Body Surface Area ◽  
Left Ventricular ◽  
Inhibitory Factor ◽  
Macrophage Migration ◽  
Endotoxin Challenge

We have recently demonstrated that macrophage migration inhibitory factor (MIF) is a myocardial depressant protein and that MIF mediates late, prolonged cardiac dysfunction after endotoxin challenge in mice. Because many factors, including endotoxin, have been implicated in the pathogenesis of cardiac dysfunction after burn injury, we tested the hypothesis that MIF might also be the mediator of prolonged cardiac dysfunction in this model. At 4 h after 40% total body surface area burn in anesthetized mice, serum MIF levels increased significantly compared with baseline (2.2-fold). This increase was accompanied by a significant decrease in cardiac tissue MIF levels (2.1-fold decrease compared with controls). This pattern was consistent with MIF release from preformed cytoplasmic stores in the heart and other organs. To determine whether MIF mediates cardiac dysfunction after burn injury, mice were pretreated with anti-MIF neutralizing monoclonal antibodies or isotype control antibodies. Beginning 4 h after burn injury (and continuing through 48 h), burned mice demonstrated a significantly depressed left ventricular shortening fraction of 38.6 ± 1.8%, compared with the normal controls (56.0 ± 2.6%). Mice treated with anti-MIF displayed an initial depression of cardiac function similar to nontreated animals but then showed rapid restoration of cardiac function with complete recovery by 24 h, which persisted for the duration of the protocol. This study is the first to demonstrate that MIF mediates late, prolonged cardiac dysfunction after burn injury and suggests that MIF blockade should be considered a therapeutic target for the treatment of burn injury.

scholarly journals Treatment of Anaemia in Patients with Acute Burn Injury: A Study of Blood Transfusion Practices

2021 ◽  
Vol 10 (3) ◽  
pp. 476
Author(s):  
Ioana Tichil ◽  
Samara Rosenblum ◽  
Eldho Paul ◽  
Heather Cleland
Keyword(s):  
Blood Transfusion ◽  
Surgical Management ◽  
Burn Injury ◽  
Total Body Surface Area ◽  
Perioperative Period ◽  
Hospital Length ◽  
Hospital Length Of Stay ◽  
Burn Care ◽  
Transfusion Requirements ◽  
Total Body

Objective: To determine blood transfusion practices, risk factors, and outcomes associated with the use of blood products in the setting of the acute management of burn patients at the Victorian Adult Burn Service. Background: Patients with burn injuries have variable transfusion requirements, based on a multitude of factors. We reviewed all acute admissions to the Victorian Adult Burns Service (VABS) between 2011 and 2017: 1636 patients in total, of whom 948 had surgery and were the focus of our analysis. Method and results: Patient demographics, surgical management, transfusion details, and outcome parameters were collected and analyzed. A total of 175 patients out of the 948 who had surgery also had a blood transfusion, while 52% of transfusions occurred in the perioperative period. The median trigger haemoglobin in perioperative was 80mg/dL (IQR = 76–84.9 mg/dL), and in the non-perioperative setting was 77 mg/dL (IQR = 71.61–80.84 mg/dL). Age, gender, % total body surface area (TBSA) burn, number of surgeries, and intensive care unit and hospital length of stay were associated with transfusion. Conclusions: The use of blood transfusions is an essential component of the surgical management of major burns. As observed in our study, half of these transfusions are related to surgical procedures and may be influenced by the employment of blood conserving strategies. Furthermore, transfusion trigger levels in stable patients may be amenable to review and reduction. Risk adjusted analysis can support the implementation of blood transfusion as a useful quality indicator in burn care.

531 The Effect of Burn Wound Size on Caloric Requirements: A Correlation of Nutritional Changes to the Clinical State

2021 ◽  
Vol 42 (Supplement_1) ◽  
pp. S113-S114
Author(s):  
Marc R Matthews ◽  
Sara Calder ◽  
Areta Kowal-Vern ◽  
Philomene Spadafore ◽  
Karen J Richey ◽  
...  
Keyword(s):  
Injury Severity ◽  
Burn Injury ◽  
Recovery Period ◽  
Total Body Surface Area ◽  
Caloric Intake ◽  
Retrospective Chart Review ◽  
Clinical State ◽  
Apache Score ◽  
Burn Wound ◽  
Caloric Requirements

Abstract Introduction Caloric intake has been a vital component for burn wound healing and recovery. The hypothesis was that caloric requirements are based on injury severity & post-burn week as predicated by indirect calorimetry (IC)/predictive equations. Methods This was a retrospective chart review of 115 burn patients (2012–2017). Caloric requirements were determined by the Curreri equation [which includes % total body surface area (TBSA)] and IC for a 5-week period provided mainly by enteral nutrition. Patients received supplements and total parenteral nutrition as needed. Results The mean ±sd age was 43±18 years, 41±18 % TBSA, Body Mass Index of 28±7 kg/m2, and mortality of 26 (23%). The major mechanisms of injury were flame/flash/explosions. There were 59 (51%) of patients with < 40 % TBSA burns, [median Injury Severity Score (ISS) 9; Apache score 14], and 56 (49%) with ≥40 % TBSA (median ISS 25; Apache score 21), p < .0001. The Respiratory Quotient (RQ) had a median of 0.94 (range 0.79 to 1.02). The median number of surgeries for the < 40 % TBSA group was 5 versus 12 for the ≥40 % TBSA, p < .0001. The Injury Factor did not differ from weeks 1–5 (1.8 for < 40 % TBSA and 2.0 for the ≥ 40 % TBSA). The Curreri equation calculation for this study was a median 3640 (range 2161–5950) calories. The Curreri equation resulted in significantly increased caloric recommendations for the ≥ 40 %TBSA compared to the < 40 %TBSA patients, p < .0001. The < 40 %TBSA group had caloric requirements ranging between 1500- 2700 calories compared to the ≥ 40 %TBSA group, whose calories ranged between 2000–3700. The total daily caloric recommendations were also significantly increased in the ≥40 %TBSA compared to the < 40 %TBSA patients. The maximum levels of resting energy expenditure (REE) from IC, total daily calories recommended by the dietitian and average calories ranged between 3000–4500 in the < 40 %TBSA group and 3600–6700 in the ≥ 40 %TBSA group. The caloric recommendations increased for all patients from week 1 to week 3 and leveled off during weeks 4–5. Conclusions Patient caloric requirements were dependent not only on the severity of the burn injury but also the post-burn hospitalization during which surgeries, debridement/grafting, and infectious complications occurred. They increased until the third week post-burn and leveled off in the recovery period. The study caloric recommendations and requirements were consistent with the REE and Curreri equation assessments.

11 Vitamin D Deficiency on Admission and the Association with Length of Stay in Patients with Thermal Injury: A Multi-center Analysis

2021 ◽  
Vol 42 (Supplement_1) ◽  
pp. S13-S14
Author(s):  
Sarah Zavala ◽  
Kate Pape ◽  
Todd A Walroth ◽  
Melissa A Reger ◽  
Katelyn Garner ◽  
...  
Keyword(s):  
Vitamin D ◽  
Length Of Stay ◽  
Vitamin D Deficiency ◽  
Burn Injury ◽  
Total Body Surface Area ◽  
Inhalation Injury ◽  
Hospital Length Of Stay ◽  
Burn Patients ◽  
Comfort Care ◽  
The Impact

Abstract Introduction In burn patients, vitamin D deficiency has been associated with increased incidence of sepsis. The objective of this study was to assess the impact of vitamin D deficiency in adult burn patients on hospital length of stay (LOS). Methods This was a multi-center retrospective study of adult patients at 7 burn centers admitted between January 1, 2016 and July 25, 2019 who had a 25-hydroxyvitamin D (25OHD) concentration drawn within the first 7 days of injury. Patients were excluded if admitted for a non-burn injury, total body surface area (TBSA) burn less than 5%, pregnant, incarcerated, or made comfort care or expired within 48 hours of admission. The primary endpoint was to compare hospital LOS between burn patients with vitamin D deficiency (defined as 25OHD < 20 ng/mL) and sufficiency (25OHD ≥ 20 ng/mL). Secondary endpoints include in-hospital mortality, ventilator-free days of the first 28, renal replacement therapy (RRT), length of ICU stay, and days requiring vasopressors. Additional data collected included demographics, Charlson Comorbidity Index, injury characteristics, form of vitamin D received (ergocalciferol or cholecalciferol) and dosing during admission, timing of vitamin D initiation, and form of nutrition provided. Dichotomous variables were compared via Chi-square test. Continuous data were compared via student t-test or Mann-Whitney U test. Univariable linear regression was utilized to identify variables associated with LOS (p < 0.05) to analyze further. Cox Proportional Hazard Model was utilized to analyze association with LOS, while censoring for death, and controlling for TBSA, age, presence of inhalation injury, and potential for a center effect. Results Of 1,147 patients screened, 412 were included. Fifty-seven percent were vitamin D deficient. Patients with vitamin D deficiency had longer LOS (18.0 vs 12.0 days, p < 0.001), acute kidney injury (AKI) requiring RRT (7.3 vs 1.7%, p = 0.009), more days requiring vasopressors (mean 1.24 vs 0.58 days, p = 0.008), and fewer ventilator free days of the first 28 days (mean 22.9 vs 25.1, p < 0.001). Univariable analysis identified burn center, AKI, TBSA, inhalation injury, admission concentration, days until concentration drawn, days until initiating supplementation, and dose as significantly associated with LOS. After controlling for center, TBSA, age, and inhalation injury, the best fit model included only deficiency and days until vitamin D initiation. Conclusions Patients with thermal injuries and vitamin D deficiency on admission have increased length of stay and worsened clinical outcomes as compared to patients with sufficient vitamin D concentrations.

scholarly journals PSIX-33 Docosahexaenoic Acid (DHA) but not Eicosapentaenoic Acid (EPA) induces the trans-differentiation of C2C12 into white-like adipocytes phenotype

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 309-310
Author(s):  
Yan Huang ◽  
Saeed Ghnaimawi ◽  
Yongjie Wang ◽  
Shilei Zhang ◽  
Jamie Baum
Keyword(s):  
Mitochondrial Biogenesis ◽  
Cultured Cells ◽  
Adipogenic Differentiation ◽  
The Body ◽  
C2c12 Cells ◽  
Brown Adipocyte ◽  
Marker Genes ◽  
Induction Medium ◽  
Spare Capacity ◽  
Epa And Dha

Abstract Muscle-derived stem cells (MDSCs, or myoblasts) play an important role in myotubes regeneration. However, these cells can differentiate into adipocytes once exposed to EPA and DHA, which are highly suggested during pregnancy. The objective of this study aims at determining the effect of isolated EPA and DHA on C2C12 cells undergoing white and brown adipogenic differentiation. Confluent cultured cells were treated with white and brown adipocyte induction medium (WIM and BIM respectively) with 50µM EPA and 50µM DHA separately. DHA treated groups differentiated into white-like adipocyte by down-regulating the expression of myogenic genes such as MyoD, MyoG, and Mrf4; but promoted white adipocyte marker genes(P < 0.05). Moreover, cells treated with WIM and DHA exhibited a decrease in mitochondrial biogenesis through suppressing PGC1a and TFAM expression (P < 0.05). Also, DHA promoted the expression of lipolysis regulating genes. DHA impaired C2C12 cells browning through reducing the mitochondrial biogenesis by significantly suppressing the expression of COX7a1, PGC1a, and UCP3 genes (P < 0.05). DHA treated groups showed an increased accumulation of lipid droplets and suppressed maximal mitochondrial respiration and spare capacity. EPA treatment reduced myogenesis regulating genes (P < 0.05) but did not affect adipogenic genes (P >0.05). Likewise, EPA suppressed the expression of WAT signature genes (P < 0.05), indicating its antagonism to DHA. EPA and WIM treatment suppressed the expression of TFAM and PGC1a, but did not affect PGC1a protein level. Although mitochondrial biogenic gene expression was reduced in EPA and BIDM treated group, no changes in mitochondrial function were observed. EPA supplementation did not affect the differential route of C2C12 into brown adipocytes. To conclude, EPA and DHA may similarly affect the integrity of muscle tissue, but DHA is a potent adipogenic and lipogenic factor that can change the metabolic profile of the body by increasing intramuscular fat.

scholarly journals Burn-induced heterotopic ossification from incidence to therapy: key signaling pathways underlying ectopic bone formation

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Xianglin Hu ◽  
Zhengwang Sun ◽  
Fengfeng Li ◽  
Chaoyin Jiang ◽  
Wangjun Yan ◽  
...  
Keyword(s):  
Growth Factor ◽  
Heterotopic Ossification ◽  
Transforming Growth Factor ◽  
Burn Injury ◽  
Hypoxia Inducible Factor ◽  
Total Body Surface Area ◽  
Adaptive Immune System ◽  
Transforming Growth Factor Β1 ◽  
Mesenchymal Transition ◽  
Endothelial To Mesenchymal Transition

AbstractBurn injury is one of the potential causes of heterotopic ossification (HO), which is a rare but debilitating condition. The incidence ranges from 3.5 to 5.6 depending on body area. Burns that cover a larger percentage of the total body surface area (TBSA), require skin graft surgeries, or necessitate pulmonary intensive care are well-researched risk factors for HO. Since burns initiate such complex pathophysiological processes with a variety of molecular signal changes, it is essential to focus on HO in the specific context of burn injury to define best practices for its treatment. There are numerous key players in the pathways of burn-induced HO, including neutrophils, monocytes, transforming growth factor-β1-expressing macrophages and the adaptive immune system. The increased inflammation associated with burn injuries is also associated with pathway activation. Neurological and calcium-related contributions are also known. Endothelial-to-mesenchymal transition (EMT) and vascularization are known to play key roles in burn-induced HO, with hypoxia-inducible factor-1 (HIF-1) and vascular endothelial growth factor (VEGF) as potential initiators. Currently, non-steroidal anti-inflammatory drugs (NSAIDs) and radiotherapy are effective prophylaxes for HO. Limited joint motion, ankylosis and intolerable pain caused by burn-induced HO can be effectively tackled via surgery. Effective biomarkers for monitoring burn-induced HO occurrence and bio-prophylactic and bio-therapeutic strategies should be actively developed in the future.

scholarly journals Pediatric burn resuscitation: past, present, and future

2017 ◽  
Vol 5 ◽  
Author(s):  
Kathleen S. Romanowski ◽  
Tina L. Palmieri
Keyword(s):  
Fluid Resuscitation ◽  
Burn Injury ◽  
Total Body Surface Area ◽  
Burn Shock ◽  
Basic Principles ◽  
Future Directions ◽  
Number Of Children ◽  
History Of ◽  
Adults And Children ◽  
Total Body

Abstract Burn injury is a leading cause of unintentional death and injury in children, with the majority being minor (less than 10%). However, a significant number of children sustain burns greater than 15% total body surface area (TBSA), leading to the initiation of the systemic inflammatory response syndrome. These patients require IV fluid resuscitation to prevent burn shock and death. Prompt resuscitation is critical in pediatric patients due to their small circulating blood volumes. Delays in resuscitation can result in increased complications and increased mortality. The basic principles of resuscitation are the same in adults and children, with several key differences. The unique physiologic needs of children must be adequately addressed during resuscitation to optimize outcomes. In this review, we will discuss the history of fluid resuscitation, current resuscitation practices, and future directions of resuscitation for the pediatric burn population.

scholarly journals Restrictive Transfusion Strategy Is More Effective in Massive Burns: Results of the TRIBE Multicenter Prospective Randomized Trial

2019 ◽  
Vol 184 (Supplement_1) ◽  
pp. 11-15 ◽  
Author(s):  
Tina L Palmieri ◽  
James H Holmes ◽  
Brett Arnoldo ◽  
Michael Peck ◽  
Amalia Cochran ◽  
...  
Keyword(s):  
Wound Healing ◽  
Burn Injury ◽  
Total Body Surface Area ◽  
Inhalation Injury ◽  
Transfusion Strategy ◽  
Restrictive Transfusion ◽  
Blood Utilization ◽  
Outcome Differences ◽  
Total Body ◽  
Restrictive Transfusion Strategy

Abstract Objectives Studies suggest that a restrictive transfusion strategy is safe in burns, yet the efficacy of a restrictive transfusion policy in massive burn injury is uncertain. Our objective: compare outcomes between massive burn (≥60% total body surface area (TBSA) burn) and major (20–59% TBSA) burn using a restrictive or a liberal blood transfusion strategy. Methods Patients with burns ≥20% were block randomized by age and TBSA to a restrictive (transfuse hemoglobin <7 g/dL) or liberal (transfuse hemoglobin <10 g/dL) strategy throughout hospitalization. Data collected included demographics, infections, transfusions, and outcomes. Results Three hundred and forty-five patients received 7,054 units blood, 2,886 in massive and 4,168 in restrictive. Patients were similar in age, TBSA, and inhalation injury. The restrictive group received less blood (45.57 ± 47.63 vs. 77.16 ± 55.0, p < 0.03 massive; 11.0 ± 16.70 vs. 16.78 ± 17.39, p < 0.001) major). In massive burn, the restrictive group had fewer ventilator days (p < 0.05). Median ICU days and LOS were lower in the restrictive group; wound healing, mortality, and infection did not differ. No significant outcome differences occurred in the major (20–59%) group (p > 0.05). Conclusions: A restrictive transfusion strategy may be beneficial in massive burns in reducing ventilator days, ICU days and blood utilization, but does not decrease infection, mortality, hospital LOS or wound healing.

scholarly journals Outcome of Acute Burn Injuries in a Tertiary Care Centre of Kathmandu Nepal

2020 ◽  
Vol 42 (2) ◽  
pp. 33-36
Author(s):  
Manish Devkota ◽  
Samit Sharma ◽  
Sangam Rayamajhi ◽  
Jayan M Shrestha ◽  
Ishwar Lohani
Keyword(s):  
Low Income ◽  
Surgical Intervention ◽  
Burn Injury ◽  
Tertiary Care ◽  
Total Body Surface Area ◽  
Burn Injuries ◽  
Low Income Countries ◽  
Female Population ◽  
Partial Thickness Burn ◽  
Flame Burn

Introduction Burn injuries are associated with higher morbidity and mortality especially in middle and low-income countries. The objective of the study is to assess the outcome of acute burn injuries in a tertiary care center of Kathmandu, Nepal. MethodsThis is a descriptive cross-sectional study of the clinical data of acute burn patients admitted from January 2016-December 2018. ResultsOut of 124 patients with burn injuries, there were more females (n=65, 52.4 %) than males (n=59, 47.6%). Flame burn was the most common mode of injury (n=71,57.3 %) followed by scald (n=22,17.7 %). Thirty-eight (30.6%) patients arrived to hospital on the same day of injury. The average time required to reach our hospital was 24 hours. The most frequently involved site was lower extremities (n=40, 32.26%) followed by upper extremities (n=33, 26.62%). Total body surface area (TBSA) involved in the burn injury ranges from 10% to 50% with a median of 15%. Hospital stay was 14 to 58 days with a median of 17 days. Partial thickness burn was seen in 114 (91.94%) patients whereas 10 (8.06%) patients had full thickness burn. Surgical intervention was needed in 71 (57.26%) patients. Among 71 patients, 12 patients underwent surgery twice. Fifty-two (41.94%) patients were managed conservatively. The mortality rate was 4.03%. ConclusionAdult female population is at high risk for burn injuries mostly due to flame burn. Delay in reaching care has also contributed for poor outcome of burn injuries. Majority of burn injury patients needed surgical intervention and hence improvement in surgical aspects can lead to better outcome of burn injuries.Keywords:

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