The Effect of Superoxide Dismutase on Inhibition of Acute Kidney Injury Induced by Sepsis Based on Kidney Tissue Histology and Murine Sepsis Score

Sepsis is one of the leading causes contributing to the incidence of acute kidney injury (AKI). Oxidative stress can be used as the main approach against sepsis-induced AKI. One of the primary antioxidants that plays a role in warding off oxidative stress is superoxide dismutase (SOD). This research aimed to observe the effect of antioxidant SOD in inhibiting sepsis in AKI based on kidney tissue histopathology. The research method was an experimental laboratory with a post-test-only control group design. Twenty-five adult male rats aged 12–16 weeks, weighing between 200 and 250 g, were randomly divided into five groups: Group I, as a positive control, where rats were injected with lipopolysaccharides (LPS); Group II, as a negative control; Group III, as treatment 1, where rats were injected with LPS and administered orally with SOD (Glisodin®) 250 IU daily; Group IV, as treatment 2, where rats were injected with LPS and administered orally with SOD (Glisodin®) 500 IU daily; and Group V, as treatment 2, where rats were injected with LPS and administered orally with SOD (Glisodin®) 1000 IU daily. Rats were administered with SOD (Glisodin®) by oral gavage with a flexible feeding tube for 16 weeks, given once daily in the morning, and then injected with LPS of 10 mg/kg body weight. Glisodin SOD had a significant effect on murine sepsis score (MSS). MSS influenced the tubular injury score linearly. We conclude that the optimal dose of SOD at 1000 IU for inhibiting sepsis-induced AKI incidence is compared to SOD at a dose of 250 and 500 IU. The antioxidant effect of SOD can prevent sepsis-induced AKI with oxidative stress events.

124. Establishment of a Post-Influenza Aspergillosis Model in Corticosteroid-Immunosuppressed Mice

Background Post-influenza aspergillosis (PIA) is a feared complication in patients with severe influenza, especially those receiving corticosteroids. However, validated murine models of PIA in a background of corticosteroid immunosuppression are lacking, compounding efforts to better characterize the immunopathology and treatment of this emerging entity. Methods 8-week-old female BALB/c mice were infected with ~5% of the lethal dose of a mouse-adapted influenza A/Hong Kong/1968 (H3N2) strain (flu), delivered by aerosolization, versus control (aerosolized saline). Mice then received two intraperitoneal injections of 10 mg cortisone acetate (CA) or mock injections on days 5 and 8 after flu infection. On day 9, mice were intranasally challenged with 50,000 A. fumigatus AF-293 conidia or mock-infected with saline. Survival was monitored until day 16 and infection severity was scored using the modified murine sepsis score (MSS, 0 = healthy to 3 = moribund). Pulmonary fungal burden was determined by an 18S quantitative PCR assay on day 16 or upon death. 15-16 mice per group were assessed across 3 independent experiments. Results Flu infection alone caused modest early morbidity, followed by full recovery of the mice until day 16. Treatment with CA after flu infection led to 12% mortality and increased morbidity that persisted until day 16 (median MSS = 0.8). Similarly, mice infected with AF after CA treatment had 12% mortality and a median MSS of 0.7. Combination of all 3 challenges (flu, CA, and AF) led to 40% mortality and severe morbidity in surviving mice (median MSS = 2.7). Likewise, prior flu infection of CA-treated, AF-infected mice increased the pulmonary fungal burden from 27k to 80k median conidial equivalents. In contrast, mice not receiving CA treatment showed consistently low morbidity (median day-16 MSS = 0.5) and minimal fungal burden after AF challenge, regardless of prior flu infection. Conclusion We have established a model of PIA in CA-immunosuppressed mice that underscores the detrimental effect of corticosteroid therapy on the outcomes of PIA. In the future, we will employ this model to study the impact of various pharmacological interventions on the natural history of PIA in the background of corticosteroid immunosuppression. Disclosures Dimitrios P. Kontoyiannis, MD, Astellas (Consultant)Cidara Therapeutics (Advisor or Review Panel member)Gilead Sciences (Consultant, Grant/Research Support, Other Financial or Material Support, Honoraria)

Sepsis of Patients Infected by SARS-CoV-2: Real-World Experience From the International HOPE-COVID-19-Registry and Validation of HOPE Sepsis Score

Background: Patients with sepsis with a concomitant coronavirus (COVID-19) infection are related to a high morbidity and mortality rate. We investigated a large cohort of patients with sepsis with a concomitant COVID-19, and we developed a risk score for the estimation of sepsis risk in COVID-19.Methods: We conducted a sub-analysis from the international Health Outcome Predictive Evaluation Registry for COVID-19 (HOPE-COVID-19-Registry, NCT04334291). Out of 5,837 patients with COVID-19, 624 patients were diagnosed with sepsis according to the Sepsis-3 International Consensus.Results: In multivariable analysis, the following risk factors were identified as independent predictors for developing sepsis: current smoking, tachypnoea (>22 breath per minute), hemoptysis, peripheral oxygen saturation (SpO2) <92%, blood pressure (BP) (systolic BP <90 mmHg and diastolic BP <60 mmHg), Glasgow Coma Scale (GCS) <15, elevated procalcitonin (PCT), elevated troponin I (TnI), and elevated creatinine >1.5 mg/dl. By assigning odds ratio (OR) weighted points to these variables, the following three risk categories were defined to develop sepsis during admission: low-risk group (probability of sepsis 3.1–11.8%); intermediate-risk group (24.8–53.8%); and high-risk-group (58.3–100%). A score of 1 was assigned to current smoking, tachypnoea, decreased SpO2, decreased BP, decreased GCS, elevated PCT, TnI, and creatinine, whereas a score of 2 was assigned to hemoptysis.Conclusions: The HOPE Sepsis Score including nine parameters is useful in identifying high-risk COVID-19 patients to develop sepsis. Sepsis in COVID-19 is associated with a high mortality rate.

DNA-Aptamer Raised against Receptor for Advanced Glycation End Products Improves Survival Rate in Septic Mice

Despite remarkable scientific advances in the understanding of molecular mechanisms for sepsis, therapeutic options are far from satisfactory. High mobility group box 1 (HMGB1), one of the ligands of receptor for advanced glycation end products (RAGE), is a late mediator of lethality in septic mice. We have recently found that the DNA-aptamer raised against RAGE (RAGE-aptamer) significantly blocks experimental diabetic nephropathy and melanoma growth and metastasis. We examined the effects of RAGE-aptamer on sepsis score, survival rate, and inflammatory and oxidative stress responses in serum, peripheral monocytes, kidneys and livers of lipopolysaccharide- (LPS-) injected mice, and on LPS-exposed THP-1 cells. RAGE-aptamer inhibited the binding of HMGB1 to RAGE in vitro. RAGE-aptamer significantly ( P = 0.002 ) improved sepsis score at 8 hours after LPS injection and survival rate at 24 hours ( P < 0.01 , 70%) in septic mice compared with LPS+vehicle- or LPS+control-aptamer-treated mice. RAGE-aptamer treatment significantly decreased expression of p-NF-κB p65, an active form of redox-sensitive transcriptional factor, NF-κB and gene or protein expression of TNF-α, IL-1β, IL-6, and HMGB1 in serum, peripheral monocytes, and kidneys of septic mice in association with the reduction of oxidative stress and improvement of metabolic acidosis, renal and liver damage. LPS-induced oxidative stress, inflammatory reactions, and growth suppression in THP-1 cells were significantly blocked by RAGE-aptamer. Our present study suggests that RAGE-aptamer could attenuate multiple organ damage in LPS-injected septic mice partly by inhibiting the inflammatory reactions via suppression of HMGB1-RAGE interaction.

Both Baicalein and Gallocatechin Gallate Efficaciously Inhibit SARS-CoV-2 Replication by Targeting Mpro and Sepsis in Mice

Severe acute syndrome coronavirus 2(SARS-CoV-2) caused the global pandemic of COVID-19 since December 2019. Although most of COVID-19’s patients are mild or common, most of the severe patients have sepsis caused by the cytokine storm, which greatly increases the case fatality rate. Moreover, there is no effective drug that can resist the novel coronavirus so far, so it’s urgent to develop antiviral drug for the SARS-CoV-2. In our research, we screened 29 compounds with a score lower than -6 from 35 flavonoid compounds by molecular docking. (-)-Gallocatechin gallate, (+)-Gallocatechin and Baicalein were identified to have potent inhibit activity with IC50 5.774±0.805μM, 13.14±2.081μM and 5.158±0.928μM by FRET assay. Subsequently, we conducted molecular docking experiments, which showed that (-)-Gallocatechin gallate, (+)-Gallocatechin and Baicalein were non-covalently bound to Mpro through π-π stacking and hydrogen bonds in the Cys145 catalytic site. We further evaluated the effect of (-)-Gallocatechin gallate and Baicalein on cytokine storm use a mouse model of sepsis. (-)-Gallocatechin gallate and Baicalein significant reduced sepsis severity based on weight, murine sepsis score and survival rate and reduced the inflammatory factors level such as TNF-α, IL-1α, IL-4 and IL-10. Overall, (-)-Gallocatechin gallate and Baicalein may be potential drugs for symptomatic treatment of COVID-19.

Diagnostic Utility of Cord Blood Serum Procalcitonin in Early Onset Neonatal Sepsis in Comparision to Sepsis Screening Parameters

Neonatal sepsis is an important cause of neonatal deaths globally. Diagnosis of neonatal sepsis is established based on microbiological tests of sepsis screen and clinical status. Mid phase markers of inflammation like CRP & Serum Procalcitonin are considered useful and sensitive for diagnosis. Most of the studies evaluating serum PCT as a diagnostic marker for neonatal septicemia have been carried out in peripheral venous blood with smaller sample sizes with inclusion of neonates without considering perinatal sepsis score. This hospital based, prospective study compares the diagnostic utility of cord blood Procalcitonin (PCT) with venous blood PCT; alone and as part of sepsis screening parameters currently in use in perinatal sepsis score positive neonates. Statistical analysis for cord blood Serum Procalcitonin (PCT) for detecting blood culture positive patients showed that PCT has a sensitivity of 44.4%, a high specificity of 86.4%, a low PPV of 33.3% and a high NPV of 91.1%. Overall diagnostic accuracy is 80.9%, indicating that cord blood PCT is a good test for identifying these patients (p < 0.05). However, venous blood PCT failed to demonstrate similar results. Conclusions- Umbilical blood sampling protects the neonates from pain of venipunctures. Cord blood PCT estimations have statistically significant correlation with blood culture and other sepsis screen parameters and better sensitivity and specificity than venous blood PCT. This early serological biomarker is valuable for the diagnostic armamentarium of neonatal septicemia for early diagnosis and management while awaiting blood culture reports and helps in reducing separation of probable sepsis neonates from mother, thus contributing in developmental supportive care.

Utility of cell-free DNA concentrations and illness severity scores to predict survival in critically ill neonatal foals

Plasma cell-free DNA (cfDNA) levels have been associated with disease and survival status in septic humans and dogs. To date, studies investigating cfDNA levels in association with critical illness in foals are lacking. We hypothesized that cfDNA would be detectable in the plasma of foals, that septic and sick-nonseptic foals would have significantly higher cfDNA levels compared to healthy foals, and that increased cfDNA levels would be associated with non-survival. Animals used include 80 foals of 10 days of age or less admitted to a tertiary referral center between January and July, 2020 were stratified into three categories: healthy (n = 34), sick non-septic (n = 11) and septic (n = 35) based on specific criteria. This was a prospective clinical study. Blood was collected from critically ill foals at admission or born in hospital for cfDNA quantification and blood culture. Previously published sepsis score (SS) and neonatal SIRS score (NSIRS) were also calculated. SS, NSIRS, blood culture status and cfDNA concentrations were evaluated to predict survival. Continuous variables between groups were compared using Kruskal-Wallis ANOVA with Dunn’s post hoc test. Comparisons between two groups were assessed using the Mann-Whitney U-test or Spearman rank for correlations. The performance of cfDNA, sepsis score and NSIRS score to predict survival was assessed by receiver operator characteristic (ROC) curve analysis including area under the curve, sensitivity and specificity using cutoffs. Plasma cfDNA was detectable in all foals. No significant differences in cfDNA concentration were detected between healthy foals and septic foals (P = 0.65) or healthy foals and sick non-septic foals (P = 0.88). There was no significant association between cfDNA and culture status, SS, NSIRS or foal survival. SS (AUC 0.85) and NSIRS (AUC 0.83) were superior to cfDNA (AUC 0.64) in predicting survival. Although cfDNA was detectable in foal plasma, it offers negligible utility to diagnose sepsis or predict survival in critical illness in neonates.

Protective Effect of Arbidol Against Pulmonary Fibrosis and Sepsis in Mice

From the perspective of epidemiology, viral immunology and current clinical research, pulmonary fibrosis may become one of the complications of patients with Coronavirus Disease 2019 (COVID-19). Cytokine storm is a major cause of new coronavirus death. The purpose of this study was to explore the effects of antiviral drug arbidol on cytokine storm and pulmonary fibrosis. Here, we use a mouse model of bleomycin-induced pulmonary fibrosis and a mouse model of fecal dilution-induced sepsis to evaluate the effects of arbidol on pulmonary fibrosis and cytokine storm. The results showed that arbidol significantly reduced the area of pulmonary fibrosis and improved lung function (reduced inspiratory resistance, lung dynamic compliance and forced vital capacity increased). Treatment with arbidol promoted reduced sepsis severity 48 h after sepsis induction, based on weight, murine sepsis score and survival rate. Arbidol observably alleviates inflammatory infiltrates and injury in the lungs and liver. Finally, we also found that arbidol reduced serum levels of pro-inflammatory factors such as TNF-α and IL-6 induced by fecal dilution. In conclusion, our results indicate that arbidol can alleviate the severity of pulmonary fibrosis and sepsis, and provide some reference for the treatment of cytokine storm and sequelae of pulmonary fibrosis in patients with COVID-19.

Cogongrass (Imperata cylindrica L.) Ethanol Extract on Sepsis Mice Model Body Weight and Sepsis Score

Sepsis causes damage for cells, behavioral phenotype regression, and will end in most patients' death. The ethanol extract of cogongrass (Imperata cylindrica L.) acts as an antioxidant. This study aimed to observe the effect of giving ECGR to body weight (BW) and the sepsis score of the sepsis mice model by lipopolysaccharide (LPS) induction. This study was an in vivo study with a randomized post-test controlled group design at the animal laboratory of Universitas Padjadjaran, 2018. We used 4 (four) groups of male mice (Mus musculus) DDY strains. Group 1 as a control, group 2: LPS 10 μL/kgBW, group 3, and 4: LPS+ECGR (90 mg/kgBW, and a dose of 115 mg/kgBW, respectively). This treatment was performed for two weeks. Every three days, we measured their body weight. After two weeks, group 2, group 3, and 4 were injected with LPS for 8 hours to induce sepsis. Next, we measured body weight and sepsis score using murine sepsis score (MSS). Then statistical analysis was performed using ANOVA and the Kruskal-Wallis test. The results showed no differences in body weight were found in the treatment groups (3 and 4) compared with control, suggesting no effect of ECGR in decreasing mice body weight. The sepsis score was more than 21 in groups treated with LPS (2, 3, and 4), suggesting LPS can induce sepsis. There was a slight decrease in scores in-group 3 and 4 compared with group 2. This study concludes that the treatment of ECGR caused no harm to body weight and slightly decreased sepsis score in the sepsis mice model. EKSTRAK ETANOL ALANG-ALANG (IMPERATA CYLINDRICA L.) TERHADAP BERAT BADAN DAN SKOR SEPSIS MENCIT MODEL SEPSISSepsis menyebabkan kerusakan sel, regresi fenotipe perilaku, dan akan berakhir kematian pada sebagian besar pasien. Ekstrak etanol akar alang-alang (Imperata cylindrica L.) (ECGR) berperan sebagai antioksidan. Penelitian ini bertujuan mengetahui pengaruh pemberian ECGR terhadap berat badan (BB) dan skor sepsis pada mencit model sepsis yang diinduksi lipopolisakarida (LPS). Penelitian ini adalah penelitian in vivo dengan desain randomized post-test controlled group di laboratoium hewan Universitas Padjadjaran tahun 2018. Kami menggunakan 4 (empat) kelompok mencit jantan (Mus musculus) strain DDY. Kelompok 1 sebagai kontrol, kelompok 2 diinduksi LPS 10 μL/kgBB, kelompok 3 dan 4 diinduksi LPS + ECGR (dosis 90 mg/kgBB dan 115 mg/kgBB masing-masing). Perlakuan ini dilakukan selama 2 minggu. Setiap tiga hari dilakukan pengukuran berat badan mencit. Setelah dua minggu, kelompok 2, kelompok 3, dan kelompok 4 diinjeksi LPS selama 8 jam untuk menginduksi sepsis. Selanjutnya, diukur berat badan dan skor sepsis menggunakan murine sepsis score (MSS). Analisis statistik menggunakan ANOVA dan Uji Kruskal-Wallis. Hasil penelitian menunjukkan tidak terdapat perbedaan berat badan pada kelompok perlakuan (3 dan 4) dibanding dengan kelompok kontrol yang menunjukkan ECGR tidak berpengaruh dalam menurunkan berat badan mencit. Skor sepsis lebih dari 21 pada kelompok yang diinduksi LPS (2, 3, dan 4) menunjukkan LPS dapat menyebabkan sepsis. Terdapat sedikit penurunan skor pada kelompok 3 dan 4 dibanding dengan kelompok 2. Simpulan penelitian ini, pengobatan ECGR tidak membahayakan berat badan dan mengakibatkan sedikit penurunan skor sepsis pada mencit model sepsis.