Impairment of μ-calpain activation by rhTNFR:Fc reduces severe burn-induced membrane disruption in the heart

AbstractStress cardiomyopathy is a major clinical complication after severe burn. Multiple upstream initiators have been identified; however, the downstream targets are not fully understood. This study assessed the role of the plasma membrane in this process and its relationship with the protease μ-calpain and tumor necrosis factor-alpha (TNF-α). Here, third-degree burn injury of approximately 40% of the total body surface area was established in rats. Plasma levels of LDH and cTnI and cardiac cell apoptosis increased at 0.5 h post burn, reached a peak at 6 h, and gradually declined at 24 h. This effect correlated well with not only the disruption of cytoskeletal proteins, including dystrophin and ankyrin-B, but also with the activation of μ-calpain, as indicated by the cleaved fragments of α-spectrin and membrane recruitment of the catalytic subunit CAPN1. More importantly, these alterations were diminished by blocking calpain activity with MDL28170. Burn injury markedly increased the cellular uptake of Evans blue, indicating membrane integrity disruption, and this effect was also reversed by MDL28170. Compared with those in the control group, cardiac cells in the burn plasma-treated group were more prone to damage, as indicated by a marked decrease in cell viability and increases in LDH release and apoptosis. Of note, these alterations were mitigated by CAPN1 siRNA. Moreover, after neutralizing TNF-α with rhTNFR:Fc, calpain activity was blocked, and heart function was improved. In conclusion, we identified μ-calpain as a trigger for severe burn-induced membrane disruption in the heart and provided evidence for the application of rhTNFR:Fc to inhibit calpain for cardioprotection.

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Topical Wound Healing Activity of Myricetin Isolated from Tecomaria capensis v. aurea

Molecules ◽

10.3390/molecules25214870 ◽

2020 ◽

Vol 25 (21) ◽

pp. 4870

Author(s):

Abdelsamed I. Elshamy ◽

Naglaa M. Ammar ◽

Heba A. Hassan ◽

Walaa A. El-Kashak ◽

Salim S. Al-Rejaie ◽

...

Keyword(s):

Wound Healing ◽

Inflammatory Cytokines ◽

Group Treatment ◽

Wound Closure ◽

Burn Injury ◽

Control Group ◽

Albino Rats ◽

Large Area ◽

Blood Capillaries ◽

Tnf Α

Wounds and burn injury are major causes of death and disability worldwide. Myricetin is a common bioactive flavonoid isolated naturally from the plant kingdom. Herein, a topical application of naturally isolated myricetin from the shoots of Tecomaria capensis v. aurea on excisional wound healing that was performed in albino rats. The wounded rats were treated every day with 10 and 20% myricetin for 14 days. During the experiment, the wound closure percentage was estimated at days 0, 7, and 14. Effects of myricetin on the inflammatory cytokines (tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and cluster of differentiation 68 (CD68) in the serum were evaluated using immunosorbent assay kits. The percentage of wound closure and contraction was delayed in wounded rats (67.35%) and was remarkably increased after treatment of wounded rats with myricetin; the treatment with 20% myricetin was the most potent (98.76%). Histological findings exhibited that 10% myricetin caused the formation of a large area of scarring at the wound enclosure and stratified squamous epithelium without the formation of papillae as in the control group. Treatment with 20% myricetin exhibited less area of scarring at the wound enclosure as well as re-epithelialization with a high density of fibroblasts and blood capillaries in the wound. Level elevations of serum pro-inflammatory cytokines, IL-1β, and TNF-α and macrophage CD68 were decreased in wounded rats treated with myricetin. Thus, it can be suggested that the enhancements in inflammatory cytokines as well as systemic reorganization after myricetin treatment may be recommended to play a crucial part in the promotion of wound healing. The findings suggest that treatment with a higher dose of myricetin was better in improving wound curing in rats. It could serve as a potent anti-inflammatory agent and can be used as an adjunctive or alternative agent in the future.

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A prospective observation on nutrition support in adult patients with severe burns

British Journal Of Nutrition ◽

10.1017/s0007114519000217 ◽

2019 ◽

Vol 121 (09) ◽

pp. 974-981 ◽

Cited By ~ 1

Author(s):

Fengmei Guo ◽

Hua Zhou ◽

Jian Wu ◽

Yingzi Huang ◽

Guozhong Lv ◽

...

Keyword(s):

Hospital Mortality ◽

Burn Injury ◽

Early Stage ◽

Energy Requirement ◽

Total Body Surface Area ◽

Nutrition Therapy ◽

Nutrition Support ◽

Burn Patients ◽

Severe Burn ◽

Severe Burns

AbstractNutrition therapy is considered an important treatment of burn patients. The aim of the study was to delineate the nutritional support in severe burn patients and to investigate association between nutritional practice and clinical outcomes. Severe burn patients were enrolled (n 100). In 90 % of the cases, the burn injury covered above 70 % of the total body surface area. Mean interval from injury to nutrition start was 2·4 (sd 1·1) d. Sixty-seven patients were initiated with enteral nutrition (EN) with a median time of 1 d from injury to first feed. Twenty-two patients began with parenteral nutrition (PN). During the study, thirty-two patients developed EN intolerance. Patients received an average of about 70 % of prescribed energy and protein. Patients with EN providing <30 % energy had significantly higher 28- d and in-hospital mortality than patients with EN providing more than 30 % of energy. Mortality at 28 d was 11 % and in-hospital mortality was 45 %. Multiple regression analysis demonstrated that EN providing <30 % energy and septic shock were independent risk factors for 28- d prognosis. EN could be initiated early in severe burn patients. Majority patients needed PN supplementation for energy requirement and EN feeding intolerance. Post-pyloric feeding is more efficient than gastric feeding in EN tolerance and energy supplement. It is difficult for severe burn patients to obtain enough feeding, especially in the early stage of the disease. More than 2 weeks of underfeeding is harmful to recovery.

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Abstract 232: PDE5A Inhibition in Burn-induced Cardiac Dysfunction

Circulation Research ◽

10.1161/res.127.suppl_1.232 ◽

2020 ◽

Vol 127 (Suppl_1) ◽

Author(s):

Jake J Wen ◽

Ravi S Radhakrishnan

Keyword(s):

Gene Expression ◽

Burn Injury ◽

Heart Function ◽

Total Body Surface Area ◽

Left Ventricular ◽

Sod Activity ◽

Heart Dysfunction ◽

Translational Study ◽

Total Antioxidant ◽

Oxidatively Modified

Severe burn injury initiates a feedback cycle of inflammation, fibrosis and oxidative stress and others that may contribute to heart dysfunction via the PDE5A-cGMP-PKG pathway. Sildenafil (SIL) has been shown decrease immune adverse events in multiple cardiac diseases by inhibition of PDE5A. In this study, using Nano LC MS/MS strategy to identify differentially expressed proteins, we will determined the pathological importance of PDE5A-dependent injury in burn-related heart dysfunction using our well-established rat model with 4 groups: sham; sham/SIL (sham with sildenafil injection); 24 hpb (60% total body surface area scald burn and harvested at 24 hours post burn) and 24 hpb/SIL (24 hpb with SIL immediately after burn). We performed proteomics, Vevo 2100 ECG, qPCR, histology and ELISA to test our hypothesis. Our results show Sildenafil 1) inhibited burn-induced PDE5A mRNA, 2) increased cGMP and PKG activity leading to recover of PKG down-regulated genes (IRAG, PLB, RGS2, RhoA and MYTP), 3) decreased the ROS level (H2O2), oxidatively modified adducts (malonyldialdehyde (MDA), carbonyls), fibrotic gene expression (ANP, BNP, COL1A2, COL3A2, aSMA and αsk-Actin), inflammation-related gene expression (RELA, IL-18 and TGF-β) from burned rats (BRs) and 4) preserved Left-ventricular heart function (CO, EF, SV, LVvol at systolic, LVPW at diastolic and FS) and restored the oxidant/antioxidant balance (total antioxidant, total SOD activity and Cu,ZnSOD activity) in BRs. We demonstrate that the PDE5A-cGMP-PKG pathway mediates burn-induced heart dysfunction and will provide the theoretic foundation to guide pre-clinical translational study of Sildenafil in burned patients.

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Calpain inhibition ameliorates scald burn-induced acute lung injury in rats

Burns & Trauma ◽

10.1186/s41038-018-0130-3 ◽

2018 ◽

Vol 6 ◽

Author(s):

Peng-Ran Du ◽

Hong-Ting Lu ◽

Xi-Xiang Lin ◽

Li-Feng Wang ◽

Yan-Xia Wang ◽

...

Keyword(s):

Acute Lung Injury ◽

Lung Injury ◽

Burn Injury ◽

Weight Ratio ◽

Dry Weight ◽

Membrane Skeleton ◽

Lung Damage ◽

Severe Burn ◽

Calpain Activity ◽

Ankyrin B

Abstract Background The molecular pattern of severe burn-induced acute lung injury, characterized by cell structure damage and leukocyte infiltration, remains unknown. This study aimed to determine whether calpain, a protease involved in both processes, mediates severe burn-induced acute lung injury. Methods Rats received full-thickness scald burns covering 30% of the total body surface area, followed by instant fluid resuscitation. MDL28170 (Tocris Bioscience), an inhibitor of calpain, was given intravenously 1 h before or after the scald burn. The histological score, wet/dry weight ratio, and caspase-3 activity were examined to evaluate the degree of lung damage. Calpain activity and its source were detected by an assay kit and immunofluorescence staining. The proteolysis of membrane skeleton proteins α-fodrin and ankyrin-B, which are substrates of calpain, was measured by Western blot. Results Time-course studies showed that tissue damage reached a peak between 1 and 6 h post-scald burn and gradually diminished at 24 h. More importantly, calpain activity reached peak levels at 1 h and was maintained until 24 h, paralleled by lung damage to some extent. Western blot showed that the levels of the proteolyzed forms of α-fodrin and ankyrin-B correlated well with the degree of damage. MDL28170 at a dose of 3 mg/kg b. w. given 1 h before burn injury not only antagonized the increase in calpain activity but also ameliorated scald burn-induced lung injury, including the degradation of α-fodrin and ankyrin-B. Immunofluorescence images revealed calpain 1 and CD45 double-positive cells in the lung tissue of rats exposed to scald burn injury, suggesting that leukocytes were a dominant source of calpain. Furthermore, this change was blocked by MDL28170. Finally, MDL28170 given at 1 h post-scald burn injury significantly ameliorated the wet/dry weight ratio compared with burn injury alone. Conclusions Calpain, a product of infiltrating leukocytes, is a mediator of scald burn-induced acute lung injury that involves enhancement of inflammation and proteolysis of membrane skeleton proteins. Its late effects warrant further study.

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Sildenafil Recovers Burn-Induced Cardiomyopathy

Cells ◽

10.3390/cells9061393 ◽

2020 ◽

Vol 9 (6) ◽

pp. 1393 ◽

Cited By ~ 3

Author(s):

Jake J. Wen ◽

Claire Cummins ◽

Ravi S. Radhakrishnan

Keyword(s):

Gene Expression ◽

Cardiac Fibrosis ◽

Burn Injury ◽

Heart Function ◽

Mrna Level ◽

Total Body Surface Area ◽

Left Ventricular ◽

Cgmp Level ◽

Sod Activity ◽

Heart Dysfunction

Background: Severe burn injury initiates a feedback cycle of inflammation, fibrosis, oxidative stress and cardiac mitochondrial damage via the PDE5A-cGMP-PKG pathway. Aim: To test if the PDE5A-cGMP-PKG pathway may contribute to burn-induced heart dysfunction. Methods: Sprague–Dawley rats were divided four groups: sham; sham/sildenafil; 24 h post burn (60% total body surface area scald burn, harvested at 24 h post burn); and 24 h post burn/sildenafil. We monitored heart function and oxidative adducts, as well as cardiac inflammatory, cardiac fibrosis and cardiac remodeling responses in vivo. Results: Sildenafil inhibited the burn-induced PDE5A mRNA level and increased the cGMP level and PKG activity, leading to the normalization of PKG down-regulated genes (IRAG, PLB, RGS2, RhoA and MYTP), a decreased ROS level (H2O2), decreased oxidatively modified adducts (malonyldialdehyde [MDA], carbonyls), attenuated fibrogenesis as well as fibrosis gene expression (ANP, BNP, COL1A2, COL3A2, αSMA and αsk-Actin), and reduced inflammation and related gene expression (RELA, IL-18 and TGF-β) after the burn. Additionally, sildenafil treatment preserved left ventricular heart function (CO, EF, SV, LVvol at systolic, LVPW at diastolic and FS) and recovered the oxidant/antioxidant balance (total antioxidant, total SOD activity and Cu,ZnSOD activity). Conclusions: The PDE5A-cGMP-PKG pathway mediates burn-induced heart dysfunction. Sildenafil treatment recovers burn-induced cardiac dysfunction.

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Correlation between Depression, Posttraumatic Stress Disorder, and Inflammatory Factors in Patients with Severe Burn Injury

The American Surgeon ◽

10.1177/000313481808400851 ◽

2018 ◽

Vol 84 (8) ◽

pp. 1350-1354 ◽

Cited By ~ 3

Author(s):

Donglin Jiang ◽

Shengyang Jiang ◽

Fang Gong ◽

Fenglai Yuan ◽

Peng Zhao ◽

...

Keyword(s):

Posttraumatic Stress Disorder ◽

Posttraumatic Stress ◽

Stress Disorder ◽

Burn Injury ◽

Self Report ◽

Inflammatory Factors ◽

Enzyme Linked Immunosorbent Assay ◽

Severe Burn ◽

Tnf Α ◽

Severe Burn Injury

We aim to investigate the relation between depression, posttraumatic stress disorder (PTSD), and inflammatory factors in patients with severe burn injury. Psychological assessment was carried out using PTSD checklist (PCL) involving a 17-item self-report questionnaire (PCL-17) and the Hamilton Rating Scale for depression (HAMD-24). The serum IL-1β, IL-6, IL-8, and tumor necrosis factor-α (TNF-α) were determined using enzyme-linked immunosorbent assay. Correlation analysis was performed to analyze the correlation between the factors and scores of PTSD and depression. Compared with the PCL-17 score, HAMD-24 score, and inflammatory factors at month 3, a significant decrease was noticed in the PCL-17 score, HAMD-24 score, and inflammatory factors at months 6 and 9 (P < 0.01). For the HAMD-24 score, significant improvements were noticed in the anxiety/somatization, cognitive disorder, blocking, sleep disorders, and depression at months 3, 6, and 9. The levels of IL-1β, IL-8, and TNF-α were positively correlated with the PCL-17 score (P < 0.05). The levels of IL-1β, IL-6, IL-8, and TNF-α were positively correlated with the HAMD-24 score (P < 0.05). Patients with severe burn injury showed obvious stress alternation displaying specific depression-related characteristics, and inflammation may involve in the pathogenesis of PTSD and depression in burn patients.

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Sodium variability is associated with increased mortality in severe burn injury

Burns & Trauma ◽

10.1186/s41038-017-0098-4 ◽

2017 ◽

Vol 5 ◽

Cited By ~ 6

Author(s):

Soman Sen ◽

Nam Tran ◽

Brian Chan ◽

Tina L. Palmieri ◽

David G. Greenhalgh ◽

...

Keyword(s):

Serum Sodium ◽

Burn Injury ◽

Total Body Surface Area ◽

Multivariate Logistic Regression Analysis ◽

Median Number ◽

High Plasma ◽

Plasma Sodium ◽

Burn Patients ◽

Severe Burn ◽

Severe Burn Injury

Abstract Background Dysnatremias are associated with increased mortality in critically ill patients. Hypernatremia in burn patients is also associated with poor survival. Based on these findings, we hypothesized that high plasma sodium variability is a marker for increased mortality in severely burn-injured patients. Methods We performed a retrospective review of adult burn patients with a burn injury of 15% total body surface area (TBSA) or greater from 2010 to 2014. All patients included in the study had at least three serum sodium levels checked during admission. We used multivariate logistic regression analysis to determine if hypernatremia, hyponatremia, or sodium variability independently increased the odds ratio (OR) for death. Results Two hundred twelve patients met entry criteria. Mean age and %TBSA for the study was 45 ± 18 years and 32 ± 19%. Twenty-nine patients died for a mortality rate of 14%. Serum sodium was measured 10,310 times overall. The median number of serum sodium measurements per patient was 22. Non-survivors were older (59 ± 19 vs. 42 ± 16 years) and suffered from a more severe burn injury (50 ± 25% vs. 29 ± 16%TBSA). While mean sodium was significantly higher for non-survivors (138 ± 3 milliequivalents/liter (meq/l)) than for survivors (135 ± 2 meq/l), mean sodium levels remained within the laboratory reference range (135 to 145 meq/l) for both groups. Non-survivors had a significantly higher median number of hypernatremic (> 145 meq/l) measurements (2 vs. 0). Coefficient of variation (CV) was significantly higher in non-survivors (2.85 ± 1.1) than survivors (2.0 ± 0.7). Adjusting for TBSA, age, ventilator days, and intensive care unit (ICU) stay, a higher CV of sodium measurements was associated with mortality (OR 5.8 (95% confidence interval (CI) 1.5 to 22)). Additionally, large variation in sodium ranges in the first 10 days of admission may be associated with increased mortality (OR 1.35 (95% CI 1.06 to1.7)). Conclusions Increased variability in plasma sodium may be associated with death in severely burned patients.

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Burn injury induces intestinal inflammatory response mediated by Th17 in burn-primed endotoxemic mice

International Surgery ◽

10.9738/intsurg-d-18-00014.1 ◽

2020 ◽

Author(s):

Kazuhiko Sekine ◽

Takayuki Shibusawa ◽

Seitaro Fujishima ◽

Naoki Aikawa ◽

Junichi Sasaki

Keyword(s):

Lamina Propria ◽

Mononuclear Cells ◽

Burn Injury ◽

Inflammatory Responses ◽

Total Body Surface Area ◽

Vital Role ◽

Mice Model ◽

Th17 Response ◽

Lamina Propria Mononuclear Cells ◽

Tnf Α

Objective: This study aimed to elucidate the mechanism underlying the susceptibility to infection-related acute lung injury by focusing on the role of gut mucosal T-helper (Th) 17 cells that preferentially produce IL-17 with probiotics in a burn-primed endotoxemic mice model. Methods: Mice were subjected to a 15% total body surface area third-degree burn. Survival from lethal lipopolysaccharide (LPS) administration (3 mg/kg) on 11th day post burn was assessed in mice fed by chow with or without 1.2% Lactobacillus powder after burn injury. Lamina propria mononuclear cells were enzymatically isolated from the ileum removed on 11th day post burn and incubated along with 1 μg/mL LPS or 10 μg/mL anti-CD3 antibody for 24 h; subsequently, the following seven cytokines were analyzed in the supernatant: IFN-γ, TNF-α, IL-2, IL-4, IL-6, IL-10, and IL-17. Results: Lactobacillus treatment post-burn injury markedly improved survival after lethal endotoxemia in burn-primed mice (64.3% vs. 21.4%, p = 0.03). The production of proinflammatory cytokines such as TNF-α, IL-6, and IL-17 by lamina propria mononuclear T-lymphocytes and macrophages including Th17 response was augmented by burn injury but decreased with Lactobacillus treatment after burn injury. Conclusions: Th17- and Th17-mediated inflammatory responses in the gut mucosa may play a vital role, which could be attenuated by Lactobacillus treatment, in survival of lethal endotoxemia in burn-primed mice.

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10 A Multi-center Study Analyzing Association of Vitamin D Deficiency and Replacement with Infectious Outcomes in Patients with Burn Injuries

Journal of Burn Care & Research ◽

10.1093/jbcr/irab032.015 ◽

2021 ◽

Vol 42 (Supplement_1) ◽

pp. S12-S13

Author(s):

Katelyn Garner ◽

Sarah Zavala ◽

Kate Pape ◽

Todd A Walroth ◽

Melissa A Reger ◽

...

Keyword(s):

Vitamin D ◽

Burn Injury ◽

Graft Loss ◽

Total Body Surface Area ◽

Inhalation Injury ◽

Control Group ◽

Comfort Care ◽

Increased Risk ◽

Multi Center Study ◽

Center Study

Abstract Introduction Vitamin D (25OHD) deficiency has been associated with poor outcomes in intensive care populations. A recent single-center, burn study found a high incidence of 25OHD deficiency. A difference was noted in infectious complications, but was underpowered. The primary objective of this multi-center study was to determine if 25OHD deficiency is associated with infectious outcomes in adult burn patients. Methods Adult patients were eligible for inclusion in this 7 center, retrospective study if admitted January 1, 2016 - July 25, 2019 and had a 25OHD concentration drawn within the first 7 days of admission. Patients were excluded if admitted for a non-burn injury, had total body surface area (TBSA) burned of less than 5%, a readmission, pregnant, incarcerated, or made comfort care or expired within 48 hours of admission. Expecting a 3:1 enrollment, goal was at least 250 total patients to be appropriately powered (β = 0.2; α = 0.05) to detect a 33% difference in composite infectious outcome (bacteremia, pneumonia, urinary tract infection, wound infection, graft loss, or death) between patients with 25OHD deficiency (< 20 ng/mL) and control (≥ 20 ng/mL). Generalized linear mixed modelling was used to control for center effect, % TBSA, age, and presence of inhalation injury and find the most predictive model. Results A total of 1147 patients were initially included. After exclusions, 234 (56.8%) in the deficient and 178 in the control group remained. Patients in the control group had their concentration drawn earlier (p < 0.001), were more likely to be male (p = 0.006), Caucasian (p < 0.001), lower body mass index (p = 0.009), lower % TBSA burn (p = 0.002), and taking a 25OHD supplement prior to admission (p < 0.001). Deficient patients were more likely to have an infectious outcome (52.1% vs 36.0%, p = 0.002), acute kidney injury requiring renal replacement therapy (p = 0.009), less ventilator free days in the first 28 days (p < 0.001), and more days requiring vasopressors (p = 0.008). After controlling for center, % TBSA, age, and inhalation injury the best model also included presence of deficiency (odds ratio = 2.425 [1.035 - 1.252]), days until 25OHD supplement initiation (1.139 [1.035 - 1.252]), and choice of cholecalciferol over ergocalciferol 2.112 [1.151 - 3.877]). Conclusions Dilution concerns were controlled by including %TBSA in the regression model. Even if low 25OHD concentrations were an acute reaction to burn injury and not representative of true deficiency, low concentrations and delay in supplementation were independently associated with increased risk of an infectious outcome.

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