Hydrogen Gas Inhalation Attenuates Endothelial Glycocalyx Damage and Stabilizes Hemodynamics in a Rat Hemorrhagic Shock Model Background: Hydrogen gas (H2) inhalation during hemorrhage stabilizes post-resuscitation hemodynamics, improving short-term survival in a rat hemorrhagic shock and resuscitation (HS/R) model. However, the underlying molecular mechanism of H2 in HS/R is unclear. Endothelial glycocalyx (EG) damage causes hemodynamic failure associated with HS/R. In this study, we tested the hypothesis that H2 alleviates oxidative stress by suppressing xanthine oxidoreductase (XOR) and/or preventing tumor necrosis factor-alfa (TNF-α)-mediated syndecan-1 shedding during EG damage. Methods: HS/R was induced in rats by reducing mean arterial pressure (MAP) to 35 mm Hg for 60 min followed by resuscitation. Rats inhaled oxygen or H2 + oxygen after achieving shock either in the presence or absence of an XOR inhibitor (XOR-I) for both groups. In a second test, rats received oxygen alone or anti-tumor necrosis factor (TNF)-α monoclonal antibody with oxygen or H2. Two hours after resuscitation, XOR activity, purine metabolites, cytokines, syndecan-1 were measured and survival rates were assessed 6 h after resuscitation. Results: H2 and XOR-I both suppressed MAP reduction and improved survival rates. H2 did not affect XOR activity and the therapeutic effects of XOR-I and H2 were additive. H2 suppressed plasma TNF-α and syndecan-1 expression; however, no additional H2 therapeutic effect was observed in the presence of anti-TNF-α monoclonal antibody. Conclusions: H2 inhalation after shock stabilized hemodynamics and improved survival rates in an HS/R model independent of XOR. The therapeutic action of H2 was partially mediated by inhibition of TNF-α-dependent syndecan-1 shedding. Address for correspondence: Motoaki Sano, M.D, PhD, 35 Shinanomachi Shinjuku-ku Tokyo 160-8582, Japan, Department of Cardiology, Keio University School of Medicine, The center for molecular hydrogen medicine, Keio University. E-mail: msano@keio.jp. Received 7 August, 2019 Revised 27 August, 2019 Accepted 27 September, 2019 Reprints will not be ordered. Conflicts of Interest and Source of Funding: The research was supported by a research grant from Taiyo Nissan Corporation to M.Sa., and xanthine oxidoreductase inhibitor (Topiroxat) was provided by Sanwa Kagaku Kenkyusho Co., Ltd. T.T, M.Sa., and M.Su. received a travel grant from the Taiyo Nippon Sanso Corporation. The remaining authors declare no conflicts of interest. Supplemental digital content is available for this article. Direct URL citation appears in the printed text and is provided in the HTML and PDF versions of this article on the journal's Web site (www.shockjournal.com). © 2019 by the Shock Society

The Shock Society 2019–2021 Strategic Plan No abstract available

Cardiac Dysfunction in Severely Burned Patients: Current Understanding of Etiology, Pathophysiology and Treatment Patients that experience severe burn injuries face a massive inflammatory response resulting in hemodynamic and cardiovascular complications. Even after immediate and appropriate resuscitation, removal of burn eschar and covering of open areas, burn patients remain at high risk for serious morbidity and mortality. As a result of the massive fluid shifts following the initial injury, along with large volume fluid resuscitation, the cardiovascular system is critically affected. Further, increased inflammation, catecholamine surge and hypermetabolic syndrome impacts cardiac dysfunction, which worsens outcomes of burn patients. This review aimed to summarize the current knowledge about the effect of burns on the cardiovascular system. A comprehensive search of the PubMed and Embase databases and manual review of articles involving effects of burns on the cardiovascular system was conducted. Many burn units use multi-modal monitors (e.g. transpulmonary thermodilution) to assess hemodynamics and optimize cardiovascular function. Echocardiography is often used for additional evaluations of hemodynamically unstable patients to assess systolic and diastolic function. Due to its non-invasive character, echocardiography can be repeated easily, which allows to follow patients longitudinally. The use of anabolic and anti-catabolic agents has been shown to be beneficial for short- and long-term outcomes of burn survivors. Administration of propranolol (non-selective β-receptor antagonist) or oxandrolone (synthetic testosterone) for up to 12 months post-burn counteracts hypermetabolism during hospital stay and improves cardiac function. A comprehensive understanding of how burns lead to cardiac dysfunction and new therapeutic options could contribute to better outcomes in this patient population. Address reprint requests to Christian Tapking, MD, MMS, Department of Surgery, University of Texas Medical Branch Galveston, 301 University Blvd, Galveston, TX 77555, USA. E-mail: christian.tapking@googlemail.com Received 21 August, 2019 Revised 11 September, 2019 Accepted 2 October, 2019 Conflicts of interest: The authors declare that they have no conflicts of interest. Funding: None. © 2019 by the Shock Society

Hemophagocytic Lymphohistiocytosis in Critically Ill Patients Background: Hemophagocytic lymphohistiocytosis (HLH), an uncontrolled overactivation of the immune system, is well characterized in pediatric patients, yet, much less is known about this life-threatening condition in adult patients. As HLH is often complicated by organ failure, patients will require admission to the intensive care unit for organ support therapy. However, recognition of HLH patients in the ICU is challenged by the clinical overlap with sepsis. Here, we analyze HLH patients to better understand its clinical presentation, diagnosis and treatment. Methods: For the purpose of this retrospective observational study, we searched for suspected and diagnosed adult HLH of all patients admitted to at least one adult surgical, anesthesiological or medical ICU between January 2006 and August 2018 at the university hospital Charité – Universitätsmedizin Berlin. All cases were reviewed by two HLH experts, who confirmed or declined the diagnosis. Results: Of 6340 ICU patients with ferritin measurement, 40 suffered from HLH (0.63%). Of these, in-hospital mortality was 60.0% over all cases, which was highest in malignancy-associated HLH (71.4%). Infections were identified as most common triggers (42.5%). A variety of 19 different treatment strategies were applied. Non-survivors showed higher ferritin at diagnosis compared to survivors (p = 0.021), which was also seen in multivariable analyses. A minimum ferritin of 4083 μg/L after diagnosis was most predictive for 30-day mortality (AUC 0.888, 95% CI 0.771–1.000; sensitivity 93.8%, specificity 78.9%). Conclusions: Mortality in adult HLH patients in the ICU is high, particularly in malignancy-associated HLH. Infections are the most frequent HLH triggers in critically ill patients. At present, there is no standardized treatment for HLH in adult patients available. Assessment of ferritin is valuable for diagnosis, prognosis, and treatment monitoring. Trial registration: The study was registered with www.ClinicalTrials.gov (NCT02854943) on August 1, 2016. Address reprint requests to Gunnar Lachmann, MD, Department of Anesthesiology and Operative Intensive Care Medicine (CCM, CVK) Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health Augustenburger Platz 1, D-13353 Berlin, Germany. E-mail: gunnar.lachmann@charite.de. Received 9 August, 2019 Revised 26 August, 2019 Accepted 17 September, 2019 Ethics approval: Ethics approval was obtained from the institutional review board (Ethikkommission der Charité – Universitätsmedizin Berlin, EA1/176/16). The study was registered with www.ClinicalTrials.gov (NCT02854943) on August 1, 2016. Consent for publication: Not applicable. Availability of data and material: Due to legal restrictions imposed by the data protection commissioner of the Charité – Universitätsmedizin Berlin, public sharing of study data with other researchers or entities is not allowed. Requests may be sent to dai-researchdata@charite.de. Competing interests: The authors declare that they have no competing interests. Funding: Gunnar Lachmann is participant of the BIH Charité Clinician Scientist Program funded by the Charité – Universitätsmedizin Berlin and the Berlin Institute of Health (BIH). Authors’ contributions: Conceived and designed the study: CK, GL. Analyzed the data: CK, FSS, GV, TSGL. Wrote the manuscript: CK, GJ, GL. Commented on the manuscript: all authors. The authors declare that they have no competing interests. Supplemental digital content is available for this article. Direct URL citation appears in the printed text and is provided in the HTML and PDF versions of this article on the journal's Web site (www.shockjournal.com). © 2019 by the Shock Society

Neonatal Sepsis Alters the Excitability of Regular Spiking Cells in the Nucleus of the Solitary Tract in Rats Objective: Sepsis is a leading cause of mortality and morbidity in infants. Although the measures of autonomic dysfunction (e.g. reduced heart rate variability) predict mortality in sepsis, the mechanism of sepsis-induced autonomic dysfunction has remained elusive. The nucleus of the solitary tract hjh(NTS) is a vital structure for the integrated autonomic response to physiological challenges. In the present study we hypothesized that sepsis alters the excitability of NTS neurons in a rat model of neonatal sepsis (14-day old rats). Methods and results: Sepsis was induced by intraperitoneal injection of cecal slurry (CS) in rat neonates. The presence of autonomic dysfunction was confirmed by observing a significant reduction in both short-term and long-term heart rate variably following CS injection. We investigated the effect of polymicrobial sepsis on the electrophysiological properties of the medial NTS neurons using a whole cell patch clamp recording. Our results showed that the resting membrane potential in regular spiking neurons was significantly less polarized in the septic group (-37.6 ± 1.76 mv) when compared with the control group (-54.7 ± 1.73 mv, P < 0.001). The number of spontaneous action potentials in the septic group, was also significantly higher than the control group (P < 0.05). In addition, the frequency and amplitude of the spontaneous excitatory post synaptic potentials (EPSPs) was significantly higher in neurons recorded in the septic group (P < 0.001). Interestingly, regular spiking cells in the CS group exhibited a rebound action potential following hyperpolarization. Injection of depolarizing currents was associated with lower first spike latency and changes in rise slope of action potential (P < 0.001). Conclusions: We showed that polymicrobial sepsis increases the excitability of regular spiking cells in the medial NTS. These alterations can potentially affect neural coding and thus may contribute to an abnormal homeostatic or allostatic physiological response to sepsis and systemic inflammation. Address reprint requests to Alireza Mani, MD, PhD, and Saeed Semnanian, MD, PhD, University College London, London, London United Kingdom. E-mail: a.r.mani@ucl.ac.uk; ssemnan@modares.ac.ir. Received 15 July, 2019 Revised 17 August, 2019 Accepted 13 September, 2019 Conflicts of interest: none Disclosure statement: Authors declare no conflict of interest. The funder had no influence on the study design or analysis or on the content of this article Funding: This study was supported by a grant from Tarbiat Modares University (PhD project grant to Golnar Eftekhari: g.eftekhari@modares.ac.ir). Supplemental digital content is available for this article. Direct URL citation appears in the printed text and is provided in the HTML and PDF versions of this article on the journal's Web site (www.shockjournal.com). © 2019 by the Shock Society

Genistein Protects Against Burn-Induced Myocardial Injury via Notch1 Mediated Suppression of Oxidative/Nitrative Stress Genistein (Gen) exhibits strong anti-oxidative/anti-nitrative activity and cardioprotective effects in several models; however, its role in burn-induced myocardial injury is unknown. This study investigated the protective effect of Gen on burn-induced myocardial injury and aimed to elucidate the mechanism of protection. Mice were injected with Gen, intraperitoneally, at different dose immediately after burn injury. The expression levels of Notch-1 intracellular domain (NICD1) and hairy and enhancer of split (Hes-1) were determined by immunoblotting. Conditional Notch-RBP-J knockout mice were used to investigate the mechanisms of Gen-induced cardioprotection. Gen alleviated burn-induced myocardial injury, as shown by improved left ventricle ejection fraction (LVEF), decreased serum LDH and CK levels, and apoptosis. Moreover, Gen decreased expressions of inducible NO synthase (iNOS) and gp91phox, reduced NO and superoxide anions production, and ameliorated their cytotoxic reaction product, peroxynitrite. More importantly, Gen significantly up-regulated the expression of NICD1 and Hes1 after burn injury. In addition, genetic knockout of Notch1 not only blocked the cardioprotection of Gen but also markedly attenuated Gen-induced anti-oxidative/anti-nitrative effect. These results demonstrate, for the first time, that Gen treatment attenuates burn-induced myocardial injury via the Notch1 mediated suppression of oxidative/nitrative stress. Address reprint requests to Dahai Hu, and Xuekang Yang, MD, Xijing Hospital Department of Burn Surgery, China. E-mail: 1047678732@qq.com; yangxkburns@163.com. Received 31 July, 2019 Revised 21 August, 2019 Accepted 1 October, 2019 Conflict of interest: None. Supplemental digital content is available for this article. Direct URL citation appears in the printed text and is provided in the HTML and PDF versions of this article on the journal's Web site (www.shockjournal.com). © 2019 by the Shock Society

Overexpression of BCL-2 in the Intestinal Epithelium Prevents Sepsis-Induced Gut Barrier Function Via Altering Tight Junction Protein Expression Sepsis induces both intestinal hyperpermeability and epithelial apoptosis. While each has been implicated in mediating sepsis mortality, the relationship between these two processes is unclear. We hypothesized that preventing intestinal apoptosis would prevent gut barrier dysfunction. To test this hypothesis, transgenic mice that overexpress the anti-apoptotic protein Bcl-2 in the gut epithelium (Fabpl-Bcl-2 mice) and wild type (WT) mice were subjected to sham laparotomy or cecal ligation and puncture and orally gavaged with fluorescein isothiocyanate conjugated-dextran (FD-4) five hours before sacrifice. Serum FD-4 concentration was assayed to measure intestinal permeability, and jejunal tight junctions were assayed for mRNA and protein expression. Baseline FD-4 concentration was similar between WT and Fabpl-Bcl-2 mice. Intestinal permeability increased 6, 12, 24 and 48 hours following sepsis in WT mice; however, FD-4 concentration was significantly lower at each timepoint in Fabpl-Bcl-2 mice. In addition, there were no statistically significant changes in permeability between septic and sham transgenic mice. Intestinal mRNA expression of claudin 3, claudin 5 and occludin were lower in septic Fabpl-Bcl-2 mice while claudin 4 mRNA levels were higher in Fabpl-Bcl-2 mice. In contrast, no differences were detected in claudins 2, 7, 15, JAM-A or ZO-1. Protein levels followed the same trend for all tight junction mediators different between WT and Fabpl-Bcl-2 mice except occludin was significantly higher in transgenic mice. Together these results demonstrate that decreasing intestinal epithelial apoptosis prevents hyperpermeability following sepsis via tight junction alterations which may be at least partially responsible for improved survival conferred by Bcl-2 overexpression. Address reprint requests to Craig Coopersmith, MD, 101 Woodruff Circle, Suite WMB 5105, Atlanta, GA 30322. E-mail: cmcoop3@emory.edu. Received 19 March, 2019 Revised 8 April, 2019 Accepted 30 September, 2019 Conflicts of Interest and Source of Funding: This work was supported by funding from the National Institutes of Health (GM072808, GM095442, GM104323, GM113228, AA027396) Supplemental digital content is available for this article. Direct URL citation appears in the printed text and is provided in the HTML and PDF versions of this article on the journal's Web site (www.shockjournal.com). © 2019 by the Shock Society

Earlier Recognition of Sepsis and Septic Shock With SEPSIS-3 Criteria – It's Still Early Days! No abstract available

Reply to the Letter to the Editor: Earlier Recognition of Sepsis and Septic Shock With Sepsis-3 Criteria – It's Still Early Days! No abstract available

Urantide Improves Cardiac Function, Modulates Systemic Cytokine Response and Increases Survival in a Murine Model of Endotoxic Shock Introduction: Urotensin II (UII) is a potent vasoactive peptide activating the G protein-coupled receptor UT, and is involved in systemic inflammation and cardiovascular functions. The aim of our work was to study the impact of the UT antagonist urantide on survival, systemic inflammation and cardiac function during endotoxic shock. Methods: C57Bl/6 mice were intraperitoneally injected with lipopolysaccharide (LPS) and then randomized to be injected either by urantide or NaCl 0.9% 3, 6 and 9 hours (H3, H6, H9) after LPS. The effect of urantide on the survival rate, the levels of cytokines in plasma at H6, H9, H12, the expression level of nuclear factor-kappa B (NF-κB-p65) in liver and kidney (at H12), and the cardiac function by trans-thoracic echocardiography from H0 to H9 was evaluated. Results: Urantide treatment improved survival (88.9% vs 30% on day 6, p < 0.05). This was associated with changes in cytokine expression: a decrease in IL-6 (2485 [2280–2751] pg/ml vs 3330 [3119–3680] pg/ml, p < 0.01) at H6, in IL-3 (1.0 [0.40–2.0] pg/ml vs 5.8 [3.0–7.7] pg/ml, p < 0.01), and IL-1β (651 [491–1135] pg/ml vs 1601 [906–3010] pg/ml, p < 0.05) at H12 after LPS administration. Urantide decreased the proportion of cytosolic NF-κB-p65 in liver (1.3 [0.9–1.9] vs 3.2 [2.3–4], p < 0.01) and kidney (0.3 [0.3–0.4] vs 0.6 [0.5–1.1], p < 0.01). Urantide improved cardiac function (left ventricular fractional shortening: 24.8 [21.5–38.9] vs 12.0 [8.7–17.6] %, p < 0.01 and cardiac output: 30.3 [25.9–39.8] vs 15.1 [13.0–16.9] ml/min, p < 0.0001). Conclusion: These results show a beneficial curative role of UT antagonism on cytokine response (especially IL-3), cardiac dysfunction and survival during endotoxic shock in mice, highlighting a potential new therapeutic target for septic patients. Address reprint requests to Hélène Castel, INSERM U1239, Laboratory of Neuronal and Neuroendocrine Cell Communication and Differentiation, University of Rouen Normandie, 25 Rue Tesnière, 76821 Mont-Saint-Aignan. E-mail: helene.castel@univ-rouen.fr Received 12 January, 2019 Revised 12 February, 2019 Accepted 6 September, 2019 Thomas Clavier and Emmanuel Besnier: These two authors contributed equally to this work. Hélène Castel and Vincent Compère: Co-last authors This work was supported by Normandie Rouen University and Inserm. Conflicts of Interest and Source of Funding: The authors have disclosed that they hold no financial interests related to the conduct or results of this research. Supplemental digital content is available for this article. Direct URL citation appears in the printed text and is provided in the HTML and PDF versions of this article on the journal's Web site (www.shockjournal.com). © 2019 by the Shock Society