Key pathways and genes that are altered during treatment with hyperbaric oxygen in patients with sepsis due to necrotizing soft tissue infection (HBOmic study).

BACKGROUND: For decades, the basic treatment strategies of necrotizing soft tissue infections (NSTI) have remained unchanged, primarily relying on aggressive surgical removal of infected tissue, broad-spectrum antibiotics, and supportive intensive care. One treatment strategy that has been proposed as an adjunctive measure to improve patient outcomes is hyperbaric oxygen (HBO2) treatment. HBO2 treatment has been linked to several immune modulatory effects; however, investigating these effects is complicated due to the disease's acute life-threatening nature, metabolic and cell homeostasis dependent variability in treatment effects, and heterogeneity with respect to both patient characteristics and involved pathogens. To embrace this complexity, we aimed to explore the underlying biological mechanisms of HBO2 treatment in patients with NSTI on the gene expression level. METHODS: We conducted an observational cohort study on prospective collected data, including 85 patients admitted to the intensive care unit (ICU) for NSTI. All patients were treated with one or two HBO2 treatments and had one blood sample taken before and after the intervention. Total RNAs from blood samples were extracted and mRNA purified with rRNA depletion, followed by whole-transcriptome RNA sequencing with a targeted sequencing depth of 20 million reads. A model for differentially expressed genes (DEGs) was fitted, and the functional aspects of the obtained set of genes was predicted with GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of genes and Genomes) enrichment analyses. All analyses were corrected for multiple testing with FDR. RESULTS: After sequential steps of quality control, a final of 160 biological replicates were included in the present study. We found 394 protein coding genes that were significantly DEGs between the two conditions with FDR < 0.01, of which 205 were upregulated and 189 were downregulated. The enrichment analysis of these DEGs revealed 20 GO terms in biological processes and 12 KEGG pathways that were significantly overrepresented in the upregulated DEGs, of which the term; "adaptive immune response" (GO:0002250) (FDR = 9.88E-13) and "T cell receptor signaling pathway" (hsa04660) (FDR = 1.20E-07) were the most significant. Among the downregulated DEGs two biological processes were significantly enriched, of which the GO term "apoptotic process" (GO:0006915) was the most significant (FDR = 0.001), followed by "Positive regulation of T helper 1 cell cytokine production" (GO:2000556), and "NF-kappa B signaling pathway" (hsa04064) was the only KEGG pathway that was significantly overrepresented (FDR = 0.001). CONCLUSIONS: When one or two sessions of HBO2 treatment were administered to patients with a dysregulated immune response and systemic inflammation due to NSTI, the important genes that were regulated during the intervention were involved in activation of T helper cells and downregulation of the disease-induced highly inflammatory pathway NF-κB, which was associated with a decrease in the mRNA level of pro-inflammatory factors. TRIAL REGISTRATION: Biological material was collected during the INFECT study, registered at ClinicalTrials.gov (NCT01790698).

Hyperbaric oxygen treatment in the management of necrotising soft-tissue infections: results from a Danish nationwide registry study.

OBJECTIVES: Application of hyperbaric oxygen (HBO2) treatment in the multidisciplinary setting of necrotising soft-tissue infection (NSTI) is debated as a considerable number of studies are of low quality with marked prognostication bias due to inadequately addressing disease severity. The objective of this study was to associate HBO2 treatment with mortality in patients with NSTI including disease severity as a prognostic variable. DESIGN: Nationwide population-based register study. SETTING: Denmark. PARTICIPANTS: Danish residents with NSTI patients between January 2011 and June 2016. PRIMARY AND SECONDARY OUTCOME MEASURES: Thirty-day mortality was compared between patients receiving and patients not receiving HBO2 treatment using inverse probability of treatment weighting and propensity-score matching with predetermined variables (age, sex and weighted Charlson comorbidity score, presence of septic shock and Simplified Acute Physiology Score II (SAPS II)). RESULTS: A total of 671 NSTI patients were included with a median age of 63 (52-71), 61% male sex, 30% had septic shock and a median SAPS II of 46 (34-58). Patients who received HBO2 treatment (n=266) were younger and had lower SAPS II, but a larger fraction had septic shock compared with patients not receiving HBO2 treatment. Overall, all-cause 30-day mortality was 19% (95% CI 17% to 23%). The statistical models were in general acceptably balanced with covariates reaching <0.1 absolute standardised mean differences and patients receiving HBO2 treatment were associated with lower 30-day mortality (OR 0.40, 95% CI 0.30 to 0.53, p<0.001). CONCLUSIONS: In analyses using inverse probability of treatment weighting and propensity score analysis, patients treated with HBO2 treatment were associated with improved 30-day survival.

Investigating the Effects of Hyperbaric Oxygen Treatment in Necrotizing Soft Tissue Infection With Transcriptomics and Machine Learning (the HBOmic Study): Protocol for a Prospective Cohort Study With Data Validation.

BACKGROUND: Necrotizing soft tissue infections (NSTIs) are complex multifactorial diseases characterized by rapid bacterial proliferation and progressive tissue death. Treatment is multidisciplinary, including surgery, broad-spectrum antibiotics, and intensive care; adjunctive treatment with hyperbaric oxygen (HBO2) may also be applied. Recent advances in molecular technology and biological computation have given rise to new approaches to infectious diseases based on identifying target groups defined by activated pathophysiological mechanisms. OBJECTIVE: We aim to capture NSTI disease signatures and mechanisms and responses to treatment in patients that receive the highest standard of care; therefore, we set out to investigate genome-wide transcriptional responses to HBO2 treatment during NSTI in the host and bacteria. METHODS: The Effects of Hyperbaric Oxygen Treatment Studied with Omics (HBOmic) study is a prospective cohort study including 95 patients admitted for NSTI at the intensive care unit of Copenhagen University Hospital (Rigshospitalet), Denmark, between January 2013 and June 2017. All participants were treated according to a local protocol for management of NSTI, and biological samples were obtained and stored according to a standard operational procedure. In the proposed study, we will generate genome-wide expression profiles of whole-blood samples and samples of infected tissue taken before and after HBO2 treatment administered during the initial acute phase of infection, and we will analyze the profiles with unsupervised hierarchical clustering and machine learning. Differential gene expression will be compared in samples taken before and after HBO2 treatment (N=85), and integration of profiles from blood and tissue samples will be performed. Furthermore, findings will be compared to NSTI patients who did not receive HBO2 treatment (N=10). Transcriptomic data will be integrated with clinical data to investigate associations and predictors. RESULTS: The first participant was enrolled on July 27, 2021, and data analysis is expected to begin during autumn 2022, with publication of results immediately thereafter. CONCLUSIONS: The HBOmic study will provide new insights into personalized patient management in NSTIs. TRIAL REGISTRATION: ClinicalTrials.gov NCT01790698; https://clinicaltrials.gov/ct2/show/NCT01790698. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/39252.

Increase in the Complement Activation Product C4d and the Terminal Complement Complex sC5b-9 Is Associated with Disease Severity and a Fatal Outcome in Necrotizing Soft-Tissue Infection.

The hyperinflammatory burden is immense in necrotizing soft-tissue infection (NSTI). The complement system is a key during the innate immune response and may be a promising target to reduce the inflammatory response, potentially improving the clinical outcome. However, complement activation and its association to disease severity and survival remain unknown in NSTI. Therefore, we prospectively enrolled patients with NSTI and sampled blood at admission and once daily for the following 3 days. Plasma C4c, C4d, C3bc, and C3dg and the terminal complement complex (TCC) were evaluated using ELISA techniques. In total, 242 patients were included with a median age of 62 years, with a 60% male predominance. All-cause 30-day mortality was 17% (95% confidence interval [CI] 13-23) with a follow-up of >98%. C4c and C3dg were negatively correlated with Simplified Acute Physiology Score II (Rho -0.22, p < 0.001 and Rho -0.17, p = 0.01). Patients with septic shock (n = 114, 47%) had higher levels of baseline TCC than those in non-shock patients (18 vs. 14, p < 0.001). TCC correlated with the Sequential Organ Failure Assessment (SOFA) score (Rho 0.19, p = 0.004). In multivariate Cox regression analysis (adjusted for age, sex, comorbidity, and SOFA score), high baseline C4d (>20 ng/mL) and the combination of high C4d and TCC (>31 arbitrary units/mL) were associated with increased 30-day mortality (hazard ratio [HR] 3.26, 95% CI 1.56-6.81 and HR 5.12, 95% CI 2.15-12.23, respectively). High levels of both C4d and TCC demonstrated a negative predictive value of 0.87. In conclusion, we found that in patients with NSTI, complement activation correlated with the severity of the disease. High baseline C4d and combination of high C4d and TCC are associated with increased 30-day mortality. Low baseline C4d or TCC indicates a higher probability of survival.

Associations between YKL-40 and markers of disease severity and death in patients with necrotizing soft-tissue infection.

BACKGROUND: Necrotizing soft-tissue infection (NSTI) is a severe and fast-progressing bacterial infection. Prognostic biomarkers may provide valuable information in treatment guidance and decision-making, but none have provided sufficient robustness to have a clinical impact. YKL-40 may reflect the ongoing pathological inflammatory processes more accurately than traditional biomarkers as it is secreted by the activated immune cells, but its prognostic yields in NSTI remains unknown. For this purpose, we investigated the association between plasma YKL-40 and 30-day mortality in patients with NSTI, and assessed its value as a marker of disease severity. METHODS: We determined plasma YKL-40 levels in patients with NSTI (n = 161) and age-sex matched controls (n = 65) upon admission and at day 1, 2 and 3. RESULTS: Baseline plasma YKL-40 was 1191 ng/mL in patients with NSTI compared with 40 ng/mL in controls (p < 0.001). YKL-40 was found to be significantly higher in patients with septic shock (1942 vs. 720 ng/mL, p < 0.001), and in patients receiving renal-replacement therapy (2382 vs. 1041 ng/mL, p < 0.001). YKL-40 correlated with Simplified Acute Physiology Score II (Rho 0.33, p < 0.001). Baseline YKL-40 above 1840 ng/mL was associated with increased risk of 30-day mortality in age-sex-comorbidity adjusted analysis (OR 3.77, 95% CI; 1.59-9.24, p = 0.003), but after further adjustment for Simplified Acute Physiology Score II no association was found between YKL-40 and early mortality. CONCLUSION: High plasma YKL-40 to be associated with disease severity, renal-replacement therapy and risk of death in patients with NSTI. However, YKL-40 is not an independent predictor of 30-day mortality.

Effect of immunoglobulin G on cytokine response in necrotising soft-tissue infection: A post hoc analysis.

BACKGROUND: A marked inflammatory response in necrotising soft-tissue infection (NSTI) may contribute to the severe clinical course. Intravenous polyspecific immunoglobulin G (IVIG) is used by some as adjuvant treatment for NSTI, but in the randomised INSTINCT trial, no effect of IVIG in NSTI patients was seen on physical quality of life. In experimental studies, IVIG may induce immunosuppressive effects by reducing the pro-inflammatory response and neutralising circulating superantigens. However, data on the potential immunomodulatory effects are sparse and remain to be investigated in a clinical setting. In this post hoc analysis of the INSTINCT trial, we aimed to assess the effect of IVIG on various inflammatory cytokines up to day 3 after randomisation. METHODS: Tumour necrosis factor (TNF), interleukin-1ß, interleukin-6, interleukin-10 and granulocyte colony-stimulating factor were measured at admission, days 1, 2 and 3. RESULTS: A total of 100 ICU patients with NSTI were included; 50 were allocated to IVIG (25 g/d for 3 days) and 50 to placebo. No difference in the overall inflammatory response was observed between groups except from TNF, which was higher in the IVIG group as compared to the placebo group (area under curve-admission to day 3, 93.6 vs 60.2, P = .02). Similarly, no differences were observed in percentage change from baseline to day 3 in any of the studied cytokines between patients allocated to IVIG group and those allocated to placebo group. CONCLUSION: In ICU patients with NSTI, IVIG did not reduce the plasma concentration of cytokines in the first 3 days.

Hyperbaric oxygen treatment impacts oxidative stress markers in patients with necrotizing soft-tissue infection.

Necrotizing soft-tissue infection (NSTI) is a rare, severe, and fast-progressing bacterial infection associated with a high risk of developing sepsis or septic shock. Increasing evidence indicates that oxidative stress is crucial in the development and progression of sepsis, but its role in NSTI specifically has not been investigated. Some patients with NSTI receive hyperbaric oxygen (HBO2) treatment as the restoration of oxidative stress balance is considered an important mechanism of action, which HBO2 facilitates. However, a gap in knowledge exists regarding the effect of HBO2 treatment on oxidative stress in patients with NSTI. In the present observational study, we aimed to investigate HBO2 treatment effects on known markers of oxidative stress in patients with NSTI. We measured plasma myeloperoxidase (MPO), superoxide dismutase (SOD), heme oxygenase-1 (HO-1) and nitrite+nitrate in 80 patients with NSTI immediately before and after their first HBO2 treatment, and on the following day. We found that HBO2 treatment was associated with a significant increase in MPO and SOD by a median of 3.4 and 8.8 ng/mL, respectively. Moreover, we observed an HBO2 treatment-associated increase in HO-1 in patients presenting with septic shock (n=39) by a median of 301.3 pg/mL. All markers were significantly higher in patients presenting with septic shock compared to patients without shock, and all markers correlated with disease severity. High baseline SOD was associated with 90-day mortality. In conclusion, HBO2 treatment was associated with an increase in MPO and SOD in patients with NSTI, and oxidative stress was more pronounced in patients with septic shock.

Hyperbaric oxygen treatment is associated with a decrease in cytokine levels in patients with necrotizing soft-tissue infection.

BACKGROUND: The pathophysiological understanding of the inflammatory response in necrotizing soft-tissue infection (NSTI) and its impact on clinical progression and outcomes are not resolved. Hyperbaric oxygen (HBO2 ) treatment serves as an adjunctive treatment; however, its immunomodulatory effects in the treatment of NSTI remains unknown. Accordingly, we evaluated fluctuations in inflammatory markers during courses of HBO2 treatment and assessed the overall inflammatory response during the first 3 days after admission. METHODS: In 242 patients with NSTI, we measured plasma TNF-α, IL-1ß, IL-6, IL-10, and granulocyte colony-stimulating factor (G-CSF) upon admission and daily for three days, and before/after HBO2 in the 209 patients recieving HBO2 . We assessed the severity of disease by Simplified Acute Physiology Score (SAPS) II, SOFA score, and blood lactate. RESULTS: In paired analyses, HBO2 treatment was associated with a decrease in IL-6 in patients with Group A-Streptococcus NSTI (first HBO2 treatment, median difference -29.5 pg/ml; second HBO2 treatment, median difference -7.6 pg/ml), and overall a decrease in G-CSF (first HBO2 treatment, median difference -22.5 pg/ml; 2- HBO2 treatment, median difference -20.4 pg/ml). Patients presenting with shock had significantly higher baseline cytokines values compared to non-shock patients (TNF-α: 51.9 vs. 23.6, IL-1ß: 1.39 vs 0.61, IL-6: 542.9 vs. 57.5, IL-10: 21.7 vs. 3.3 and G-CSF: 246.3 vs. 11.8 pg/ml; all p < 0.001). Longitudinal analyses demonstrated higher concentrations in septic shock patients and those receiving renal-replacement therapy. All cytokines were significantly correlated to SAPS II, SOFA score, and blood lactate. In adjusted analysis, high baseline G-CSF was associated with 30-day mortality (OR 2.83, 95% CI: 1.01-8.00, p = 0.047). CONCLUSION: In patients with NSTI, HBO2 treatment may induce immunomodulatory effects by decreasing plasma G-CSF and IL-6. High levels of inflammatory markers were associated with disease severity, whereas high baseline G-CSF was associated with increased 30-day mortality.

Adjunctive hyperbaric oxygen treatment for necrotising soft-tissue infections: A systematic review and meta-analysis.

INTRODUCTION: Surgical intervention, broad-spectrum antibiotics and intensive care support are the standard of care in the treatment of necrotising soft-tissue infections (NSTI). Hyperbaric oxygen treatment (HBOT) may be a useful adjunctive treatment and has been used for almost 60 years, but its efficacy remains unknown and has not been systematically appraised. The aim was to systematically review and synthesise the highest level of clinical evidence available to support or refute the use of HBOT in the treatment of NSTI. METHODS: The review was prospectively registered (PROSPERO; CRD42020148706). MEDLINE, EMBASE, CENTRAL and CINAHL were searched for eligible studies that reported outcomes in both HBOT treated and non-HBOT treated individuals with NSTI. In-hospital mortality was the primary outcome. Odds ratio (ORs) were pooled using random-effects models. RESULTS: The search identified 486 papers of which 31 were included in the qualitative synthesis and 21 in the meta-analyses. Meta-analysis on 48,744 patients with NSTI (1,237 (2.5%) HBOT versus 47,507 (97.5%) non-HBOT) showed in-hospital mortality was 4,770 of 48,744 patients overall (9.8%) and the pooled OR was 0.44 (95% CI 0.33-0.58) in favour of HBOT. For major amputation the pooled OR was 0.60 (95% CI 0.28-1.28) in favour of HBOT. The dose of oxygen in these studies was incompletely reported. CONCLUSIONS: Meta-analysis of the non-random comparative data indicates patients with NSTI treated with HBOT have reduced odds of dying during the sentinel event and may be less likely to require a major amputation. The most effective dose of oxygen remains unclear.

Soluble ICAM-1 is modulated by hyperbaric oxygen treatment and correlates with disease severity and mortality in patients with necrotizing soft-tissue infection.

The inflammatory response in patients with necrotizing soft-tissue infection (NSTI) is excessive and often causes collateral damage, thereby worsening disease severity and prognosis. Shedding of endothelial adhesion molecules may be a key regulatory mechanism to modulate the inflammatory response in patients with septic NSTI. Hyperbaric oxygen (HBO2) treatment has demonstrated an effect on adhesion molecules. However, endothelial shedding and its association with NSTI disease severity and prognosis is not fully understood. We hypothesized that shedding of intercellular adhesion molecule-1, and the resulting release of the soluble isoform soluble intercellular adhesion molecule-1 (sICAM-1), is modified by HBO2 treatment, and that sICAM-1 concentrations are associated with severity of disease and mortality in patients with NSTI. We measured sICAM-1 in 80 patients with NSTI immediately before and after first session of HBO2 treatment as well as on the following day. We found an overall sICAM-1 level of 594 ng/mL [interquartile range (IQR) 406-817]. HBO2 significantly (P = 0.01) increased sICAM-1 by a median of 45.1 ng/mL, which remained elevated until the following day; this effect was more pronounced in patients with septic shock. Furthermore, sICAM-1 was significantly correlated with disease severity [simplified acute physiology score II (SAPS II); ρ = 0.24, P = 0.04] and low sICAM-1 was found to be an independent predictor for 90-day mortality in age-sex-SAPS II-adjusted analysis (odds ratio 14.0, 95% CI 1.82-341.4, P = 0.03). These results support the hypothesis that endothelial shedding is an important pathophysiological mechanism in NSTI and suggest that HBO2 treatment may induce immunomodulatory effects that potentially decreases collateral damage and mortality.NEW & NOTEWORTHY HBO2 treatment may be a promising immunomodulatory agent by increasing sICAM-1, thereby lowering risk of collateral damage, especially in the most critically ill patients. sICAM-1 is associated with disease severity in NSTI as emphasized by significant correlations with SAPS II. Low sICAM-1 levels are an independent risk factor of 90-day mortality and appeared to give a good level of diagnostic accuracy, suggesting that sICAM-1 can be used as a prognostic biomarker for NSTI.

Incidence, comorbidity and mortality in patients with necrotising soft-tissue infections, 2005-2018: a Danish nationwide register-based cohort study.

OBJECTIVE: To assess the incidence, comorbidities, treatment modalities and mortality in patients with necrotising soft-tissue infections (NSTIs) in Denmark. DESIGN: Nationwide population-based registry study. SETTING: Denmark. PARTICIPANTS: Danish residents with NSTI between 1 January 2005 and 31 August 2018. MAIN OUTCOME MEASURE: Incidence of disease per 100 000 person/year and all-cause mortality at day 90 obtained from Danish National Patient Registry and the Danish Civil Registration System. RESULTS: 1527 patients with NSTI were identified, yielding an incidence of 1.99 per 100 000 person/year. All-cause 30-day, 90-day and 1-year mortality were 19.4% (95% CI 17.4% to 21.5%), 25.2% (95% CI 23.1% to 27.5%) and 30.4% (95% CI 28.0% to 32.8%), respectively. Amputation occurred in 7% of the individuals. Diabetes was the most predominant comorbidity affecting 43% of the cohort, while 26% had no comorbidities. Higher age, female sex and increasing comorbidity index were found to be independent risk factors of mortality. Admission to high-volume hospitals was associated with improved survival (OR 0.59, 95% CI 0.45 to 0.77). Thirty-six per cent received hyperbaric oxygen therapy (HBOT) as an adjunctive therapy. No change in overall mortality was found over the studied time period. CONCLUSION: The present study found that in Denmark, the incidence of NSTI increased; mortality rates remained high and largely unaltered. Diabetes was the most common comorbidity, while higher age, female sex and increasing comorbidity index were associated to increased mortality. Survival was improved in those admitted to hospitals with more expertise in treating NSTI. In high-volume hospital, HBOT was associated with decreased odds for mortality.

[Hyperbaric oxygen therapy].

Hypoxia triggers hypoxia-inducible factor (HIF). Not only hypoxia triggers downstream HIF target genes for transcription, as intermittent hyperoxia also possesses similar capabilities, suggesting that fluctuations in oxygen availability may be equally important for inducing HIF transcription. This review describes some of the mechanisms, whereby intermittent hyperbaric hyperoxia may explain some of the observations during hyperbaric oxygen therapy such as enhanced wound healing, angiogenesis and tissue healing, and concludes that oxidative stress enhances certain antibiotics in infection control.

Measurement of peripheral arterial tonometry in patients with diabetic foot ulcers during courses of hyperbaric oxygen treatment.

INTRODUCTION: Treatment of diabetic foot ulcers is complex and often protracted. Hyperbaric oxygen treatment (HBOT) improves wound healing in diabetic ulcers and serves as an important adjunct to regular diabetic wound care. Endothelial dysfunction plays a central role in diabetes-related vascular complications and may be evaluated by a non-invasive technique called peripheral arterial tonometry which measures a reactive hyperaemia index (RHI). We hypothesized that endothelial function measured by peripheral arterial tonometry is impaired in diabetic foot ulcer patients and that HBOT might improve endothelial function. METHODS: Endothelial function was prospectively assessed by peripheral arterial tonometry in 22 subjects with diabetic foot ulcers and 17 subjects without diabetes during courses of HBOT. Endothelial function was evaluated before first (baseline) and 30th treatments, and at 90-day follow-up. Serum insulin growth factor-I (IGF-I) concentrations were determined by immunoassay. Results were compared to 23 healthy subjects. RESULTS: No baseline differences were found in endothelial function between subjects with diabetes, HBOT patients without-diabetes and healthy control subjects (RHI; 1.26, 1.61 and 1.81, respectively). No significant changes in RHI were found in patients with (P = 0.17) or without (P = 0.30) diabetes during courses of HBOT. At 90-day follow-up IGF-I was significantly reduced in the subjects with diabetes (P = 0.001) and unchanged in the group without diabetes (P = 0.99). CONCLUSIONS: We found no significant differences in RHI between subjects with diabetic foot ulcers and patients without diabetes, nor improvement in endothelial function assessed by peripheral arterial tonometry during courses of HBOT.

Patient's characteristics and outcomes in necrotising soft-tissue infections: results from a Scandinavian, multicentre, prospective cohort study.

PURPOSE: Necrotising soft-tissue infections (NSTI) are characterised by necrosis, fast progression, and high rates of morbidity and mortality, but our knowledge is primarily derived from small prospective studies and retrospective studies. METHODS: We performed an international, multicentre, prospective cohort study of adults with NSTI describing patient's characteristics and associations between baseline variables and microbiological findings, amputation, and 90-day mortality. RESULTS: We included 409 patients with NSTI; 402 were admitted to the ICU. Cardiovascular disease [169 patients (41%)] and diabetes [98 (24%)] were the most common comorbidities; 122 patients (30%) had no comorbidity. Before surgery, bruising of the skin [210 patients (51%)] and pain requiring opioids [172 (42%)] were common. The sites most commonly affected were the abdomen/ano-genital area [140 patients (34%)] and lower extremities [126 (31%)]. Monomicrobial infection was seen in 179 patients (44%). NSTI of the upper or lower extremities was associated with monomicrobial group A streptococcus (GAS) infection, and NSTI located to the abdomen/ano-genital area was associated with polymicrobial infection. Septic shock [202 patients (50%)] and acute kidney injury [82 (20%)] were common. Amputation occurred in 22% of patients with NSTI of an extremity and was associated with higher lactate level. All-cause 90-day mortality was 18% (95% CI 14-22); age and higher lactate levels were associated with increased mortality and GAS aetiology with decreased mortality. CONCLUSIONS: Patients with NSTI were heterogeneous regarding co-morbidities, initial symptoms, infectious localisation, and microbiological findings. Higher age and lactate levels were associated with increased mortality, and GAS infection with decreased mortality.

SuPAR correlates with mortality and clinical severity in patients with necrotizing soft-tissue infections: results from a prospective, observational cohort study.

Necrotizing soft tissue infections (NSTI) have a 90-day mortality rate of 18-22%. Tools are needed for estimating the prognosis and severity of NSTI upon admission. We evaluated soluble urokinase-type plasminogen activator receptor (suPAR) levels at admission as a prognostic marker of NSTI severity and mortality. In a prospective, observational cohort study, suPAR was measured in 200 NSTI patients. We compared admission suPAR levels in survivors and non-survivors, patients with septic shock and non-shock, amputation and non-amputation, correlations with Simplified Acute Physiology Score II (SAPS II) and the Sequential Organ Failure Assessment (SOFA) score. Admission suPAR levels were higher in septic shock vs. non-septic shock patients (9.2 vs. 5.8 ng/mL, p-value < 0.001) and non-survivors vs. survivors (11 vs. 6.1 ng/mL, p-value < 0.001) and correlated with SAPS II (r = 0.52, p < 0.001) and SOFA score (r = 0.64, p < 0.001). Elevated suPAR upon admission was associated with 90-day mortality (log-rank test p < 0.001), however not after adjustment for age, sex, and SOFA score. The AUC for suPAR and 90-day mortality was 0.77. We found that suPAR is a promising candidate for prognosis and severity in patients with NSTI.

Neuronal nitric oxide inhibition attenuates the protective effect of HBO2 during cyanide poisoning.

PURPOSE: Experiments have shown that hyperbaric oxygen (HBO2) therapy reduces cyanide-induced cerebral distress. The exact mechanism behind HBO2's neuroprotective effect is unknown, but has been proposed to be mediated by an increased neuronal nitric oxide (NO) bioavailability, which may compete with cyanide for the active site of cytochrome oxidase in the mitochondrial respiratory chain. We hypothesized that the ameliorating effect of HBO2 is caused by an increased bioavailability of NO, which can be attenuated by injection of the selective neuronal NO synthase inhibitor, 7-nitroindazole, preceding the HBO2 procedure. METHODS: A total of 41 anesthetized female Sprague-Dawley rats were allocated to four groups: 1) vehicle [1.2 ml isotonic NaCl via intra-arterial administration]; 2) cyanide [5.4 mg/kg potassium CN (KCN) intra-arterial] plus 7-nitroindazole [25 mg/kg 7-nitroindazole via intraperitoneal injection]; 3) cyanide plus 7-nitroindazole plus HBO2 [284 kPa for 90 minutes]; 4) cyanide plus 7-nitroindazole plus normobaric oxygen [101.3 kPa for 90 minutes]. Cerebral interstitial lactate, glucose, glycerol and pyruvate were evaluated by means of microdialysis. RESULTS: HBO2 during inhibition of nNOS worsened cerebral metabolism compared to both solely CN-intoxicated animals and normobaric oxygen-treated animals. This was indicated by elevated lactate (in mM; 0.85 vs. 0.63 and 0.42, P=0.006 and P ⟨ 0.001, respectively), glycerol (in mM; 46 vs. 17 and 14, both P ⟨ 0.001), glucose (in mM; 0.58 vs. 0.31 and 0.32, both P ⟨ 0.001). CONCLUSIONS: The results indicate that a specific nNOS inhibition offsets the ameliorating effect of HBO2 during cerebral CN intoxication. However, other factors might contribute to this neuroprotective effect as well.

Decompression illness treated in Denmark 1999-2013.

INTRODUCTION: The incidence, diver characteristics and symptomatology of decompression illness (DCI) in Denmark has not been assessed since 1982, and the presence of long-term residual symptoms among divers receiving hyperbaric oxygen therapy in Denmark has never been estimated to our knowledge. METHODS: We undertook a retrospective study of the incidence and characteristics of DCI cases in Denmark for the period of 1999 to 2013. Medical records and voluntary questionnaires were reviewed, extracting data on age, gender, weight, height, diver certification level, diving experience, number of previous dives, type of diving, initial type of hyperbaric treatment and DCI symptoms. Trend in annual case numbers was evaluated using run chart analysis and Spearman's correlation. Age, height, weight, and BMI were evaluated using linear regression. The presence of long-term residual symptoms was investigated by phone interviewing the subgroup of divers treated in 2009 and 2010. RESULTS: Two-hundred-and-five DCI cases were identified. The average annual case load was 14 with no significant trend during the study period (P = 0.081). Nor did we find any trend in age, weight, height or BMI. The most frequent symptoms were paraesthesia (50%), pain (42%) and vertigo (40%). Thirteen out of the subgroup of 30 divers had residual symptoms at discharge from hospital, and six out of 24 of these divers had residual symptoms at the time of follow-up. CONCLUSIONS: We observed a more than ten-fold increase in DCI-cases since the period 1966-1980. In the subgroup of divers treated in 2009/2010, a quarter had long-term residual symptoms as assessed by telephone interview, which is in keeping with the international literature, but still a reminder that DCI can have life-long consequences.

Postural stability in patients with decompression sickness evaluated by means of Quantitative Romberg testing.

OBJECTIVE: The present study was designed to retrospectively evaluate the use of quantitative Romberg's testing on postural stability during the course of hyperbaric oxygen (HBO2) therapy in patients presenting with decompression sickness (DCS). METHODS: The Quantitative Romberg test was used to evaluate postural stability in 33 patients with DCS treated between May 2009 and August 2014. Postural stability was assessed before and after each session of HBO2 therapy. Patients were allocated into groups according to whether they presented with vertigo or not. RESULTS: Significantly higher sway values obtained with the Quantitative Romberg test were observed in the group of DCS with vertigo relative to DCS without vertigo and healthy controls. A stepwise improvement in postural instability for DCS patients with vertigo was found following HBO2 therapy. After three treatments of HBO2, postural stability was found to be within the normal range of healthy controls. CONCLUSIONS: The Quantitative Romberg test offers the the clinician a fast, reliable and objective set of parametrical data to document postural instability in patients with either confirmed or suspected DCS.

Evaluation of endothelial function by peripheral arterial tonometry and relation with the nitric oxide pathway.

Endothelial dysfunction is an important component in the development of cardiovascular diseases. Endothelial function may be evaluated by peripheral arterial tonometry (PAT) which measures the vasodilator function in the microvasculature of the fingertip during reactive hyperaemia. The reactive hyperaemia index (RHI) is decreased in the presence of cardiovascular risk factors and thus far several studies have shown that PAT-RHI may provide reliable prediction of outcome. The technique is operator independent and easy to perform. Abnormalities measured by PAT follow the same trend as those measured by flow-mediated dilation in the brachial artery, but the two methods are not interchangeable. We have reviewed the recent literature in an effort to evaluate peripheral arterial tonometry as a method to assess the function of the endothelium and additionally suggest directions for future research. Special attention will be directed to the nitric oxide dependency of the reactive hyperaemia index obtained by peripheral arterial tonometry.