The biomarkers sRAGE and d f in combination are significant predictors of mortality in COVID-19 patients

Introduction: COVID-19 infection is associated with marked inflammatory response and the patients who are admitted to the hospital are at increased risk of developing venous thromboembolism. sRAGE (soluble receptor for advanced glycation end-products) are acutely elevated in host inflammatory response to infections [1]. Fractal dimension ( df), the biomarker of clot microstructure that measures thrombogenicity has shown to be elevated in acute inflammatory conditions such as sepsis and severe sepsis. The aim of the study was to analyse these biomarkers in COVID-19 infection and whether these biomarkers help to predict mortality. Method(s): 120 suspected COVID-19 patients were recruited from the Emergency Department of a tertiary teaching hospital. One patient was excluded because they were anticoagulated, blood samples were taken to perform fractal dimension ( df) and sRAGE. Result(s): When compared to PCR -ve group, 95 patients in the PCR + ve group had significantly elevated sRAGE (p < 0.001), but not df (p = 0.43). When compared to those who survived, sRAGE was significantly elevated (p = 0.01) in 14 patients who died in PCR + ve group, but not df (p = 0.08). No significant correlation existed between sRAGE levels and df in those patients who survived (p = 0.72) or died (p = 0.92). Logistic regression analysis showed that sRAGE and df in combination acted as highly significant predictors of mortality in COVID-19 (p = 0.009) in PCR + ve group. Conclusion(s): COVID-19 patients had a profound inflammatory response as evidenced by significantly elevated sRAGE levels. This inflammatory process was more profound in those who died. The thrombogenicity in COVID-19 patients and those who died with COVID-19 appears to be not significant as measured by df. sRAGE in combination with df can be utilised as significant predictors of mortality in COVID-19 patients.

Oleic acid-induced acute lung injury in Ob/Ob mice fed with high-fat-diet exacerbate bile acid uptake and liver injury that was reversed by antagonizing their entry

Background and aims: Acute respiratory distress syndrome (ARDS) is a serious complication of COVID-19 and present in a large percentage of COVID-19 deaths. Many studies suggest that people with obesity are at increased risk of severe COVID-19, however, mechanism on liver-lung axis remains unknown. We aimed to evaluate whether bile acid (BAs) trafficking interfere with acute lung injury (ALI) in animal model with obesity. Method: Leptin deficient (ob/ob) mice fed with high-fat-diet (Ob/Ob HFD) were i.p injected with oleic acid (OA) to induce ALI. To modulate BAs uptake, mice were i.p treated with neutralizing antibody for sodium taurocholate co-transporting polypeptide (NTCP;BAs-transporter). Broncho-alveolar lavage fluid (BALF), lungs, livers and serum were obtained from mice and assessed for inflammatory (HandE staining, ALT and pro-inflammatory panel of cytokines), fibrosis (Sirius red staining, a-smooth muscle actin, collagen and fibronectin) and metabolic (BAs, cholesterol, triglyceride, glucose tolerance test (GTT) and fasting blood sugar (FBS)) profiles. In addition, alveolarcapillary membrane injury of surfactant D (SP-D) and the receptor for advanced glycation end-products (RAGE). BAs trafficking were assessed in primary lung cells and their impact on proliferation and apoptosis were evaluated. Results: Compared to WT-littermates, OA-induced lung injury and was worsened in the in the Ob/Ob HFD in the histopathology outcome. In addition, BALF of the Ob/Ob HFD showed elevated levels of BAs (3- fold;P = 0.002) associated with increased GM-CSF, INF-g, IL-1, IL-6 and IL-8 (p < 0.01). Moreover, Ob/Ob HFD with OA showed elevated serum levels in liver enzymes, lipids, glucose and metabolic markers (p < 0.01). In addition, Ob/Ob HFD livers showed an exacerbated fibrosis profile. NTCP neutralizing antibody in Ob/Ob HFD while inhibited BAs uptake/trafficking in both primary alveolar type II (BALF showed 4-fold increase in BAs) and primary hepatocytes (serum showed 3-fold increase in BAs). SP-D, RAGE and serum metabolic markers were suppressed to normal in line with enhance lung and liver histology and maintaining cell viability. Conclusion: Modulation of BAs trafficking from the liver of obese mice to the lungs could be an important step in the pathogenesis of ALI. Antagonizing BAs uptake may suggest a therapeutic strategy in improving liver-lung axis.

Network Pharmacology, Molecular Docking and Molecular Dynamics Simulation Studies of the Molecular Targets and Mechanisms of ChuanKeZhi in the Treatment of COVID-19

Objectives: Coronavirus disease 2019 (COVID-19) has had a global impact and is spreading quickly. ChuanKeZhi injection (CKZI) is widely used in asthma patients. In this paper, we aimed to explore active compounds of CKZ and determine potential mechanisms against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) through network pharmacology, molecular docking and dynamic simulation studies. Materials and Methods: We used the Systematic Pharmacology Database and Analysis Platform of Traditional Chinese Medicine (TCMSP) to screen active compounds and potential target proteins of CKZ. COVID-19 target genes were screened via the American National Center for Biotechnology Information (NCBI) gene database and human gene database (GeenCards). The protein interaction network was constructed by the Protein Interaction Network Database (Search Tool for the Retrieval of Interacting Genes/Proteins (STRING)) platform. GO enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed by the Metascape database. The main active compounds of CKZ were docked with angiotensin-converting enzyme 2 (ACE2), spike protein S1, and SARS-CoV-2-3CL pro and also docked with hub targets. We performed molecular dynamics (MD) simulation studies for validation. Results: We finally obtained 207 CKZ potential targets and 4681 potential COVID-19 targets. Key targets included mainly AKT1, TNF, IL6, VEGFA, IL1B, TP53, JUN, CASP3, etc. There were 217 Gene Ontology (GO) items in the GO enrichment analysis (p < 0.05). The main KEGG pathways included the advanced glycation end products (AGE)- receptor for AGE (RAGE) signalling pathway in diabetic complications, rheumatoid arthritis, chemical carcinogenesis-receptor activation, alcoholic liver disease, etc. Molecular docking and dynamics simulation studies both exhibited great binding capacity. Conclusions: Network pharmacology, molecular docking and dynamics simulation studies were used to identify the potential and key targets, pharmacological functions, and therapeutic mechanisms of CKZI in the treatment of COVID-19. CKZI may be an effective and safe drug in COVID-19 treatment. However, further work is needed for validation.

Social isolation promotes mammary cancer recurrence, increases IL6/JAK/STAT3 signaling and causes mitochondrial dysfunction in tamoxifen treated rats, and a traditional herbal mixture reverses all these changes

Social isolation is associated with increased risk and mortality from many diseases, such as breast cancer. Socially isolated breast cancer survivors have a 43% higher risk of recurrence and a 64% higher risk of breast cancer-specific mortality than socially integrated survivors. Since Covid-19 has dramatically increased the incidence of social isolation, it is important to determine if social isolation affects the response to endocrine therapy and/or recurrence after the therapy is completed. Since previous studies indicate that social isolation increases circulating inflammatory cytokines, we investigated if an anti-inflammatory herbal mixture Jaeumkanghwa-tang (JGT) prevents the adverse effects of social isolation on breast cancer mortality. Estrogen receptor positive mammary tumors were initiated with 7,12-dimethylbenz[a]anthracene. When a rat developed a palpable mammary tumor, it was either socially isolated (SI) by housing it singly or a rat was allowed to remain group-housed (GH). Tamoxifen (340ppm via diet) or tamoxifen + JGT (500ppm via drinking water) started when the first mammary tumor reached a size of 11 mm in diameter. Tamoxifen administration ended when a complete response to this therapy had lasted for 9 weeks (corresponds to 5 years in women). During tamoxifen therapy, social isolation non-significantly reduced the rate of complete responses to 21%, from 31% in GH group (p>0.05). After the therapy was completed, SI significantly increased local mammary tumor recurrence (p<0.001;45% GH vs 75% SI). RNAseq analysis was performed in the mammary glands. Gene set enrichment analysis (GSEA) of transcriptome showed that the increased recurrence risk in socially isolated rats was associated with an enrichment of IL6/JAK/STAT3 signaling: this result was confirmed in the tumors. In addition, oxidative phosphorylation (OXPHOS) pathway was suppressed: the suppressed genes included those involved in mitochondrial pyruvate transport and conversion of pyruvate to acetyl CoA as well as genes in the TCA cycle and mediating electron transport in mitochondrial complexes I-IV. Social isolation also increased the expression of inflammatory receptor for advanced glycation end-products (RAGE) (p≤0.05). Consumption of an anti-inflammatory JGT inhibited IL6/JAK/STAT3 signaling, upregulated OXPHOS signaling and prevented the increased risk of mammary cancer recurrence in socially isolated animals. The percentage of recurrences in the SI rats dropped from 75% without JGT to 22% with JGT (p<0.001). Breast cancer mortality among socially isolated survivors may be most effectively prevented by focusing on the period following endocrine therapy using tools that inhibit IL6/JAK/STAT3 inflammatory cytokine signaling and correct disrupted OXPHOS and mitochondrial dysfunction.

Prospective Evaluation of Serial Protein Biomarker Measurements in Sepsis-Induced Acute Respiratory Distress Syndrome

RATIONALE: Protein biomarkers including soluble receptor for advanced glycation end-products (sRAGE), angiopoietin-2 (Ang- 2), and surfactant protein-D (SP-D) have been studied for diagnosis or prognostication in acute respiratory distress syndrome (ARDS). However, prior studies of ARDS biomarkers often included heterogeneous populations, and rarely examined serial measurements. Our aim was to determine the association between serially measured sRAGE, Ang-2, and SP-D levels and ARDS development in patients with sepsis. METHODS: Adult patients admitted to the medical ICU at Grady Memorial Hospital within 72 hours of sepsis diagnosis were enrolled into this prospective observational cohort study within 48 hours of ICU admission. Patients who already had ARDS at the time of screening were excluded. After obtaining informed consent, serial plasma samples were collected on day 1, 2, and 3 of enrollment, and were analyzed for sRAGE, Ang-2, and SP-D levels using ELISA. The patients were followed for up to 28 days for relevant clinical characteristics and outcomes. The primary outcome was ARDS development according to the Berlin Definition. The secondary outcome was mortality. Pulmonary sepsis was defined as the primary infection being pneumonia (including COVID19) or aspiration pneumonia. The biomarker levels and their changes from day 1 to days 2/3 were compared between those who developed ARDS versus those who did not. RESULTS: Among 80 patients with sepsis enrolled between September 1, 2020 and June 22, 2021, 15 patients (18.8%) developed ARDS and 65 patients (81.3%) did not. ARDS patients had higher proportions of pulmonary sepsis (14/15 [93.3%] vs. 30/65 [46.2%], p=0.001) and COVID19 (7/15 [46.7%] vs. 7/65 [10.8%], p=0.003) compared to non-ARDS patients. ARDS patients had higher SP-D levels on days 1 and 2, and had a greater increase in sRAGE levels from day 1 to day 3, compared to non-ARDS patients (Figure 1A- 1B). Within the ARDS group, those who died had higher sRAGE levels on day 1 compared to those who survived (Figure 1C). CONCLUSIONS: In this analysis, ARDS patients had higher SP-D and a greater increase in sRAGE over time compared to non- ARDS patients. Non-survivors of ARDS also had higher sRAGE compared to survivors. Our findings suggest that early serial biomarker measurements may be useful for identifying sepsis patients at risk of developing ARDS and adverse clinical outcomes, and for risk stratifying sepsis patients in ARDS clinical trials focused on early therapeutics and prevention. Larger studies are needed for more detailed analyses and confirmation of these findings.