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1.
Gastroenterology ; 162(7):S-886, 2022.
Article in English | EMBASE | ID: covidwho-1967381

ABSTRACT

Endothelial cells (ECs) lining the blood vessels of all organs express the SARS-CoV2 receptor. In the absence of preexisting tissue damage, the virus would need to pass through the ECs to blood vessels to infect other tissues. Thus, EC are a target for SARS-CoV-2 infection and a conduit for viral dissemination to distant organs. We hypothesized that ECs infection and/ or injury are the mechanisms of COVID-19 pathology and multi-organ dissemination and injury. Methods: Human studies: We used lung, heart, kidney, and small bowel specimens obtained during autopsies (n=5) from COVID-19 patients and uninfected subjects. Studies: 1) histologic evaluation of endothelial damage and endotheliitis, 2) immunohistochemistry for vWF, PAI-1, VCAM-1, & ICAM-1. Studies in cultured human microvascular ECs (HMVECs): We cultured lung and cardiac HMVECs in the presence or absence of SARSCoV- 2 S1 and/or S2 protein (10 ng/ml) for 0 - 24 hr. Studies:1) cell viability and proliferation;2) angiogenesis on Matrigel and cell migration;3) mitochondrial membrane potential (MMP);4) RNA seq analysis;5) Western blotting for vWF, PAI-1, VCAM-1, and ICAM-1. We examined the protective effect of melatonin, Coenzyme Q10 and nerve growth factor on S1/S2 protein induced HMVEC cell damage. Results: Histopathologic examination revealed presence of endothelial abnormalities and endotheliitis with marked presence of inflammatory cells in vessel wall & lumen, and fibrinous microthrombi) in lung, heart & kidney in autopsy specimens of COVID-19 patients. Immunostaining visualized increased vWF, PAI-1, VCAM- 1, & ICAM-1 in COVID-19. In in vitro study, S1 and S2 proteins induced endothelial injury, reduced angiogenesis and phosphorylated/activated Erk and Akt proteins in cultured HMVECs. Treatment of HMVECs for 1 & 4 hours with S2 but not S1 protein increased ICAM-1 levels by 1.4- to 1.8-fold (P < 0.001). RNA Seq analysis showed that treatment of HMVECs with S1 and S2 proteins upregulated VCAM-1, ICAM-1 and E-selectin mRNA in cultured HMVECs. Melatonin, Coenzyme Q10 and NGF stimulated angiogenesis in HMVECs by 2.4-, 1.3-&1.4-fold (all P < 0.001). Conclusions: 1) Significant endothelial abnormalities, blood vessel damage and endotheliitis are present in lung, heart and kidney autopsy specimens of COVID-19 patients, 2) There is increased expression of vWF, PAI-1, VCAM-1, and ICAM- 1 in lung, heart, and kidney specimens of COVID-19 patients, 3) Treatment of cultured HMVECs with SARS-CoV-2 S1 and S2 proteins upregulates VCAM-1, ICAM-1 and Eselectin expression, 4) SARS-CoV-2 S1 and S2 proteins induce endothelial injury in cultured HMVECs, and 5) melatonin, Coenzyme Q10 and NGF stimulated EC function. These studies uncovered novel mechanism – endothelial dysfunction underlying SARS-CoV-2 and identified melatonin, Coenzyme Q10 and NGF as potential drugs for treatment of COVID- 19-induced EC injury

2.
Gastroenterology ; 162(7):S-247, 2022.
Article in English | EMBASE | ID: covidwho-1967258

ABSTRACT

Background: Gastric muscularis propria immune cells play an instrumental role in homeostasis and disease. A subset of these cells, muscularis macrophages (MMs) are involved in the pathobiology of diabetic gastroparesis (DG) but are poorly understood. This study aims to survey transcriptional and functional profiling of gastric MMs in DG and diabetes. Methods: Full-thickness gastric body biopsies were obtained from patients with DG and diabetic controls. CD45+ cells were isolated from dissociated muscle tissue using magnetic beads. 10xGenomics was used for scRNA-seq library prep and cells sequenced by Illumina HiSeq4000. Bioinformatic analyses was performed using Suite and Seurat. Myeloid cells were annotated through a pseudogating strategy that identifies cells by differential expression levels of HLA-DR, CD14, CD11b, and CD11c based on flow cytometry-based gating utilized in a recent analysis of human small intestinal MMs. Canonical signaling pathways were determined using Ingenuity Pathway Analysis (IPA). Results: A total of 21,740 high-quality single-cell transcriptomes were generated from 16 subjects (DG=6, age 32±8 yr, BMI 23.7±3.9, 48.2±40.1% 4 hr gastric retention, average GCSI score 3.7±0.5;Diabetic controls= 10, age 53±13 yr, BMI 42.2±5.7). Through annotating 8,693 myeloid cells (DG 1509, Controls 7184), we characterized 1,788 as MMs (CD45+HLA-DR+) and 448 as dendritic cells (CD14-CD11c+). Utilizing a priori markers for pseudogating, the MMs were divided into four populations (Figure 1): subset 1 (CD14+CD11c+HLA-DRint, 5.6%), subset 2 (CD14+CD11c+HLA-DRhi, 36.0%), subset 3 (CD14+CD11c-CD11b-, 41.8%), and subset 4 (CD14+CD11c-CD11b+, 16.6%). The overall proportions of cells in the 4 subsets were similar to a prior approach in small bowel using gating. The expected ratio of cells from DG/diabetic control was 21% based on imputed cells. Subsets 1 and 4 were significantly decreased in DG compared to controls with ratios 15% and 14% respectively while subsets 2 and 3 were unchanged (21% and 20%). On IPA, phagosome formation and immune cell trafficking represented canonical signaling pathways of subset 1 and coronavirus phagocytosis pathway and phagosome formation of subset 4. Canonical genes of subset 1 included S100A12, A8, A9, and CSTA and subset 4 as LYVE1, MAF, MRC1 (CD206), MS4A4, and A2M. Subset 4 also had the highest expression of neuron-related genes (NPTX2, BMP2) similar to that observed in the small intestine. Conclusions: Pseudogating based on the transcriptomic expression of gastric immune cells reveal MM clusters similar in gene expression and proportions to previously characterized MMs in human small bowel using gating. The reduction of MM clusters associated with anti-inflammatory, phagocytosis, and neuronal signaling in specialized MMs subsets may suggest candidate targets in the pathophysiology of DG. Supported by NIHDK074008. (Figure Presented) Figure 1. Single-Cell RNA-Seq Profiling of Human Gastric Muscularis Macrophages in DG and Diabetes. T-distributed Stochastic Neighbor Embedding (tSNE) plot of muscularis macrophages in DG and diabetic control subjects by their differential genes from MAST (FDR < 0.05), color-coded by Status. *Mf1 and Mf2 not visualized as distinct clusters due to inadequate separation of overall gene expression in cells distinguished by HLA-DRint (Mf1) and HLA-DRhi (Mf2)

3.
Gastroenterology ; 162(7):S-68-S-69, 2022.
Article in English | EMBASE | ID: covidwho-1967239

ABSTRACT

Introduction: Gut dysbiosis is associated with immune dysfunction and severity in COVID- 191-2. This study aimed to determine targeting dysbiosis as a therapy and its effect on antibody formation, gut dysbiosis and immune profile in patients with COVID-19. Material & Methods: In an open-label study, 25 consecutive hospitalized patients with COVID- 19 received a novel microbiome immunity formula (SIM01) for 28 days;30 patients who did not receive the intervention acted as controls. We collected fecal and blood samples at baseline and week 5 and followed subjects from admission up to five weeks. We performed multi-omics analysis using data from peripheral blood mononuclear cell (PBMC) transcriptome, fecal metagenomic sequencing and fecal metabolomic profiling (Figure 1A). Results: Significantly more COVID-19 patients on SIM01 developed anti-SARS-CoV-2 IgG than the control group at 2 weeks (Figure 1B). Patients on SIM01 (but not controls) showed a significant reduction of plasma levels of interleukin (IL)-6, macrophage colony-stimulating factor (M-CSF), tumour necrosis factor (TNF-a), IL-1RA (Figure 1C) and downregulated COVID-19 related signalling pathway in PBMC at Week 5. Fecal samples of subjects on SIM01 were enriched in commensal bacteria and reduced in opportunistic pathogens at week 4 and 5. Elevated plasma acetic acid in SIM01 group showed a negative correlation with SARS-CoV-2 viral load in nasopharyngeal samples (Figure 2A). Increased relative abundance of Bifidobacteria adolescentis and Coprococcus comes in fecal samples in SIM01 group positively correlated with plasma acetic acid levels (Figure 2B). Conclusion: We showed for the first time a novel microbiome formula SIM01 was effective in hastening antibody formation against SARS-CoV-2, reduced pro-inflammatory immune markers and restored gut dysbiosis in hospitalised COVID-19 patients. References: 1. Zuo T, Zhang F, Lui GCY, et al. Alterations in gut microbiota of patients with COVID-19 during time of hospitalization. Gastroenterology 2020;159:944-955 e8. 2. Yeoh YK, Zuo T, Lui GC, et al. Gut microbiota composition reflects disease severity and dysfunctional immune responses in patients with COVID- 19. Gut 2021;70:698-706. (Figure Presented) (Figure Presented)

4.
Biomedicine and Pharmacotherapy ; 153, 2022.
Article in English | EMBASE | ID: covidwho-1966381

ABSTRACT

Dexamethasone acts as an immunosuppressive drug and has been used recently in the management of specific coronavirus disease 2019 (COVID-19) cases;however, various adverse effects could limit its use. In this work, we studied the mitigation effects of black pepper oil (BP oil) on glycemic parameters, dyslipidemia, oxidative and nitrosative stress and pancreatic fibrosis in dexamethasone-treated rats. Animals were divided into five groups that were treated with vehicle, dexamethasone (10 mg/kg, SC) or black pepper oil (BP oil, 0.5 mL, or 1 mL/kg) or metformin (50 mg/kg) plus dexamethasone for 4 consecutive days. Serum insulin, blood glucose, total cholesterol, triglycerides, and Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) were higher in the dexamethasone group vs the control group and decreased in BP oil and metformin groups relative to the dexamethasone group. Pancreatic nitric oxide, inducible nitric oxide synthase and malondialdehyde levels were increased in the dexamethasone group vs the control group and decreased in BP oil and metformin groups relative to the dexamethasone group. Pancreatic endothelial nitric oxide synthase and reduced glutathione were declined in the dexamethasone group vs the control group. They were increased in BP oil and metformin groups relative to the dexamethasone group. Moreover, the pancreatic islets diameter and collagen deposition were assessed and found to be higher in the dexamethasone group vs the control group. BP oil and metformin groups showed to regress this effect. In conclusion, BP oil may alleviate hyperglycemia, hyperinsulinemia, insulin resistance, dyslipidemia and pancreatic structural derangements and fibrosis by suppressing oxidative stress, increasing endogenous antioxidant levels, modulating nitric oxide signaling, preventing pancreatic stellate cells transition and collagen deposition.

5.
Indian Journal of Pharmaceutical Sciences ; 84(3):617-630, 2022.
Article in English | EMBASE | ID: covidwho-1957666

ABSTRACT

Drug repositioning may be a promising way to find potential therapies against coronavirus disease 2019. Although chloroquine and hydroxychloroquine showed controversial results against the coronavirus disease 2019 disease, the potential common and diverging mechanisms of action are not reported and need to be dissected for better understanding them. An integrated strategy was proposed to systematically decipher the common and diverging aspects of mechanism of chloroquine and hydroxychloroquine against coronavirus disease 2019-disease network based on network pharmacology and in silico molecular docking. Potential targets of the two drugs and coronavirus disease 2019 related genes were collected from online public databases. Target function enrichment analysis, tissue enrichment maps and molecular docking analysis were carried out to facilitate the systematic understanding of common and diverging mechanisms of the two drugs. Our results showed that 51 chloroquine targets and 47 hydroxychloroquine targets were associated with coronavirus disease 2019. The core targets include tumor necrosis factor, glyceraldehyde 3-phosphate dehydrogenase, lymphocyte-specific protein-tyrosine kinase, beta-2 microglobulin, nuclear receptor coactivator 1, peroxisome proliferator-activated receptor gamma and glutathione disulfide reductase. Both chloroquine and hydroxychloroquine had good binding affinity towards tumor necrosis factor (affinity=-8.6 and -8.4 kcal/mol, respectively) and glyceraldehyde 3-phosphate dehydrogenase (-7.5 and -7.5 kcal/mol). Chloroquine and hydroxychloroquine both had good affinity with angiotensin-converting enzyme 2, 3-chymotrypsin-like protease and transmembrane serine protease 2. However, hydroxychloroquine manifested better binding affinity with the three proteins comparing with that of chloroquine. Chloroquine and hydroxychloroquine could have potential to inhibit over-activated immunity and inflammation. The potential tissue-specific regulation of the two drugs against severe acute respiratory syndrome coronavirus 2 infection may related with the lung, liver, brain, placenta, kidney, blood, eye, etc. In conclusion, our data systematically demonstrated chloroquine and hydroxychloroquine may have potential regulatory effects on coronavirus disease 2019 disease network, which may affect multiple organs, protein targets and pathways. Routine measurements of the chloroquine and hydroxychloroquine blood concentrations and tailored therapy regimen may be essential. But, further rigorous and high quality randomized controlled clinical trials are warranted to validate the antiviral effects of chloroquine and hydroxychloroquine against severe acute respiratory syndrome coronavirus 2. Our proposed strategy could facilitate the drug repurposing efforts for coronavirus disease 2019 treatment.

6.
Biomedical Reviews ; 32:37-46, 2021.
Article in English | EMBASE | ID: covidwho-1957608

ABSTRACT

Butyrylcholinesterase (BChE), a hepatic enzyme produced by the liver is affected by and influences a variety of inflammatory, infectious and metabolic dysfunction processes. Considering that COVID-19 is a multisystem disorder related to conditions influenced by BChE, the potential interrelation of the two is reviewed. BChE is altered in a variety of infectious diseases, and serves as a prognostic marker in both infections and in non-infectious diseases. Closely related to acetylcholinesterase (AChE), BChE plays a role in modulating inflammation via the cholinergic system. It forms part of the signaling pathway linking the immune system, nervous system and the endocrine system. COVID-19 progresses to a stage of unregulated inflammation in a subset of subjects. Cholinergic dysfunction could be potentially responsible for a march to cytokine storm. BChE could influence the course of COVID-19 by acting through the brain-immune-endocrine axis via cholinergic transmission, as well as affecting factors predicting adverse outcomes of COVID-19 (obesity, insulin resistance, coronary artery disease, type 2 diabetes mellitus). Interestingly, variant forms of the enzyme with impaired hydrolytic activity are reported in endogamous ethnic populations. It would be instructive to study the effect of COVID-19 in these natural human knock-out equivalents. Biomed Rev 2021;32: 37-46.

7.
Nevrologiya, Neiropsikhiatriya, Psikhosomatika ; 14(2):91-97, 2022.
Article in Russian | EMBASE | ID: covidwho-1957599

ABSTRACT

The novel coronavirus infection is commonly referred to as COVID-19, sometimes by the name of the causative agent of a respiratory viral infection, as SARS-CoV-2. Frequently, the course of COVID-19 is divided into three main periods: acute COVID-19 (up to 4 weeks), post-acute COVID-19 (from 4 to 12 weeks), post-COVID (post-COVID;from 12 weeks to 6 months). If a more protracted course of COVID (over 6 months) is discussed, the term “long-COVID” is used. All observations demonstrated a high incidence of pain syndromes of various localization in the post-COVID period and long-COVID. According to survey data, 92.3% of patients with COVID-19 reported musculoskeletal problems at the time of admission. Pain syndrome is observed in 56.3% of cases 1 month after hospitalization. Three months after COVID-19, myalgia was observed in 40.55% of cases, joint pain in 39.18%, back pain in 31.62%, and lower back pain in 24.74%. After 6 months, joint pain continues to be observed in 18.59% of patients, myalgia - in 15.09%, back pain - in 14.39%, lower back pain - in 11.23%. In 50.8% of cases, patients reported new-onset pain, of which 38.5% had pain of moderate severity (≥3 points on the visual analog scale). Patients with new-onset pain during COVID had worse quality of life indicators and a negative correlation with the pain syndrome severity, which significantly hampered recovery. Data from a meta-analysis that included 47,910 patients with long-COVID and with a protracted course of COVID indicate that 19% of them had pain in the joints of various localization. The direct cytopathic effect of SARS-CoV-2 and the systemic immune inflammation that occurs in response to infection cause damage to the joint tissue. According to the Guidelines for the Treatment of Patients with the Consequences of COVID-19, it is recommended to use slow acting structure-modifying drugs - SYSADOA - in the pharmacological treatment regimen for patients with osteoarthritis, among which parenteral forms of pharmaceutically standardized drugs - chondroitin sulfate (CS) and glucosamine sulfate (GS) are preferred. GS and CS are inhibitors of the signaling cascade of the nuclear factor NF-κB, which is involved in the realization of biological effects of a pro-inflammatory cytokine (tumor necrosis factor α), the excessive activity of which is associated with the cytokine storm in COVID-19.

8.
Natural Product Communications ; 17(7), 2022.
Article in English | EMBASE | ID: covidwho-1956964

ABSTRACT

Objective: The Chinese herbal formula Huo-Xiang-Zheng-Qi (HXZQ) is effective in preventing and treating coronavirus disease 19 (COVID-19) infection;however, its mechanism remains unclear. This study used network pharmacology and molecular docking techniques to investigate the mechanism of action of HXZQ in preventing and treating COVID-19. Methods: The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) was used to search for the active ingredients and targets of the 10 traditional Chinese medicines (TCMs) of HXZQ prescription (HXZQP). GeneCards, Online Mendelian Inheritance in Man (OMIM), Pharmacogenomics Knowledge Base (PharmGKB), Therapeutic Target Database (TTD), and DrugBank databases were used to screen COVID-19-related genes and intersect them with the targets of HXZQP to obtain the drug efficacy targets. Cytoscape 3.8 software was used to construct the drug-active ingredient–target interaction network of HXZQP and perform protein–protein interaction (PPI) network construction and topology analysis. R software was used to perform Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Finally, AutoDock Vina was utilized for molecular docking of the active ingredients of TCM and drug target proteins. Results: A total of 151 active ingredients and 250 HXZQP targets were identified. Among these, 136 active ingredients and 67 targets of HXZQP were found to be involved in the prevention and treatment of COVID-19. The core proteins identified in the PPI network were MAPK1, MAPK3, MAPK8, MAPK14, STAT3, and PTGS2. Using GO and KEGG pathway enrichment analysis, HXZQP was found to primarily participate in biological processes such as defense response to a virus, cellular response to biotic stimulus, response to lipopolysaccharide, PI3K-Akt signaling pathway, Th17 cell differentiation, HIF-1 signaling pathway, and other signaling pathways closely related to COVID-19. Molecular docking results reflected that the active ingredients of HXZQP have a reliable affinity toward EGFR, MAPK1, MAPK3, MAPK8, and STAT3 proteins. Conclusion: Our study elucidated the main targets and pathways of HXZQP in the prevention and treatment of COVID-19. The study findings provide a basis for further investigation of the pharmacological effects of HXZQP.

9.
Sexually Transmitted Infections ; 98:A44, 2022.
Article in English | EMBASE | ID: covidwho-1956918

ABSTRACT

Introduction The Covid-19 pandemic has dramatically accelerated the point of care (POC) landscape, increasing awareness and demand for rapid diagnostics of other diseases. STIs are a major current health issue, with Neisseria gonorrhoea (NG) and Chlamydia trachomatis (CT) being highly prevalent. Current diagnostic methods are not POC and the most rapid takes around 20 mins. We have developed a new molecular assay, taking < 10 minutes for a diagnostic result, and combined it with a novel, rapid detection mechanism to produce a fully integrated POC device. Methods Our assay will make use of the exponential amplification reaction (EXPAR), a rapid isothermal DNA amplification technique, to produce an output detectable by Linear Dichroism (LD). LD is a highly sensitive optical detection technique, relying on exploiting structural properties of a scaffold such as M13 bacteriophage. Adapting EXPAR and combining it with a DNA sensitive LD assay allows detection of specific DNA sequences, signalling the presence of CT and/or NG. A principal advantage of our system is it allows multiplexing on the same detection scaffold. Results Initial clinical trials using EXPAR show detection of CT/NG patient samples within 10 minutes of DNA amplification at a constant temperature. The sequences detected represent specific and well conserved regions of CT and NG. Large signal changes give M13 a high analytical sensitivity. Discussion Our systems will ensure faster and more accurate diagnosis and ultimately better patient health outcomes. We are currently focused on an expansion of the scope of diseases that we can detect, using our in-house sequence selection process. (Figure Presented).

10.
Cardiovascular Research ; 118:i19, 2022.
Article in English | EMBASE | ID: covidwho-1956561

ABSTRACT

Background and purpose: Increased inflammatory cytokines, including interleukin 6 (IL-6), are associated to enhanced arrhythmogenic risk, including atrial fibrillation [1]. Moreover, direct effects of cytokines on ion channels are emerging as important mediators of arrhythmogenic remodeling [2]. In line with this, enhanced arrhythmogenesis in COVID-19 patients is hypothesized to be driven by cytokine storms, a well demonstrated condition in this setting [3]. To dissect the underlying mechanisms explaining such an association, we evaluated the proarrhythmogenic alterations of IL-6, assessing the impact on the expression of Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels, of the regulatory subunits MiRP1, and on the action potential (AP) profile in HL-1 cardiomyocytes (CMs). In human left atrial samples we studied the relation occurring between the expression levels of IL-6 and of HCN channels. In human induced pluripotent stem cell (hiPSC)-derived CMs we evaluated the acute effects of IL-6 on pacemaker activity. Methods: HL-1 CMs exposed to 50 ng/ml IL-6 or vehicle were collected after 0, 0.5, 6, 12, 24 and 48 h to study intracellular signaling, ion channel expression and AP profile. The latter was assessed through a high-throughput system allowing optical detection of APs with optical stimulation. In human atrial samples obtained from patients undergoing surgery, IL-6 and HCN mRNA expression were analyzed by quantitative RT-PCR. The acute effects on pacemaker activity were evaluated in hiPSC-derived CMs exposed to increasing concentrations of IL-6. Results: In HL1 CMs IL-6 rapidly induces STAT3 phosphorylation, demonstrating the activation of IL-6 signaling cascade. IL-6 modifies HCN channel transcript and proteins at different time points, evidencing a significant downregulation of HCN4 isoform and significant upregulations of HCN1, HCN2 and MiRP1. In line with this, in human left atrial samples, expression levels of IL-6 were linearly and directly related to HCN1 channel, while they were linearly and inversely related to HCN4. Electrophysiological recordings on HL-1 CMs showed a decreasing trend of AP amplitude and of maximum diastolic potential, while AP durations tended to increase. In hiPSC-derived CMs IL-6 reduces the frequency of AP in a concentration-dependent manner. Conclusions: Our data demonstrate that in HL-1 CMs IL-6 activates a STAT3 dependent intracellular signaling that is associated to subsequent variation of HCN channel expression and a concurrent alteration of AP profile. The relation between IL-6 and HCN1,4 expression in human samples suggests a mechanistic link between IL-6 levels and ionic channel targets, including HCN channels. The reduction of AP frequency in hiPSC-derived CMs suggests a direct interaction with ion channels. We hypothesize that these modifications may lay the basis to enhance the propensity of atria to develop arrhythmias in condition of elevate IL-6 levels.

11.
Sleep Science ; 15:34, 2022.
Article in English | EMBASE | ID: covidwho-1935129

ABSTRACT

Introduction: Social isolation, as a result of the coronavirus pandemic, favored the exposure of elderly people to stressors that may have compromised the quality of sleep. Objective: To verify the quality of sleep of elderly people in the face of the COVID-19 pandemic. Methods: A study was carried out with 305 elderly from 61 cities, recruited through social networks, from February- June/2021. The online questionnaire included general data (city, age, gender, skin, color, education level). Questions were asked about the practice of Physical Activity (PA), the quality of sleep during the pandemic (Mini Sleep Questionnaire/MSQ altered sleep score ≥ 25;EPWORTH-sleepiness score ≥ 10 points) and whether was diagnosed with COVID-19. Results: Out of 305 individuals who participated, 71% were between 60 to 69 years, and 72% were women. In addition, 59.5% practiced some type of physical activity, 9.5% contracted the SARS-CoV2 virus, 65% reported that their sleep pattern was not altered by the pandemic, 28.8% had impaired sleep quality, 72% were not taking sleep medication. The sleepiness was within the normal range. The relation of sleep quality of elderly practitioners of PA did not show statistically significant difference in relation to those who did not practice it during the pandemic. Sleep quality and the diagnosis of COVID-19 did not present relevant statistical data either. Although the individuals did not show any change in sleep quality during the pandemic, the data signaled a poor quality of sleep in the elderly surveyed, with a score showing an overall mean (MSQ) of 31.7 indicating severe difficulty in sleeping. In this study, women had a higher sleep quality score (32.6) than men (29.5), signaling the worsening of sleep in females. Elderly people in age group 70 years more demonstrated a better sleep pattern than age group 60 to 69 years. There was a statistically significant difference in the quality of sleep of the elderly who snored (33.1) compared to those who did not snore (28.0) and those who took a nap during the day (32.7) and those who did not take a nap (30,4). Tiredness was also shown to be a sign of worsening sleep quality when related to those who were not tired during the day. Conclusion: The results suggest that there were no significant changes in the sleep pattern of the elderly during the period evaluated in the pandemic. The changes found were similar in the literature for this population.

12.
Open Access Macedonian Journal of Medical Sciences ; 10:1383-1391, 2022.
Article in English | EMBASE | ID: covidwho-1939099

ABSTRACT

BACKGROUND: No gold standard therapy was approved globally for COVID-19 pneumonia to the date of this study. The pathophysiology of SARS-CoV-2 infection displayed the predominance of hyperinflammation and immune dysregulation in inducing multiorgan damage. Therefore, the potential benefits of both immune modulation and suppression in COVID-19 have been extensively discussed as a modality to control cytokine release syndrome (CRS). Abnormally high levels of interleukin-6 (IL-6) are a common finding in COVID-19 patients with pneumonia and acute respiratory distress syndrome, so the use of IL-6 antagonist was tested as a therapeutic option in controlling the disease. Tocilizumab is a recombinant humanized anti-human IL-6 receptor monoclonal antibody that can specifically bind the membrane-bound IL-6 receptor and soluble IL-6 receptor, thereby inhibiting signal transduction. Tocilizumab is currently FDA approved for the management of rheumatoid arthritis, giant cell arthritis, polyarticular juvenile idiopathic arthritis, and systemic juvenile idiopathic arthritis. This study is a retrospective analysis of data polled during Phase I of COVID pandemic, adopted by the isolation hospital of Kasr Al-Ainy Medical School, Cairo University, during the period from May to September 2020. AIM: The aim of this study is to evaluate tocilizumab influence in the outcome;in terms of reducing the hospital stay, risk and duration of mechanical ventilation (invasive and noninvasive), mortality, and the incidence of complications related to drugs use (secondary bacterial infection and GIT bleeding) in patients with moderate-to-severe COVID-19. METHODS: This retrospective, observational cohort study included adults (between 18 and 80 years) with moderate-to-severe COVID-19 pneumonia, who were admitted to isolation hospital of Kasr Al-Ainy Medical School, Cairo University, between May and September 2020. We segregated the patients into two groups: Group A: In addition to the standard care protocol according to the local guidelines of the Egyptian Ministry of Health and Population in that period (supplemental oxygen, steroids in a dose of 1–2 mg/kg methylprednisolone for 5–10 days, broad-spectrum antibiotics, vitamins, and prophylactic dose of anticoagulation with low-molecular-weight heparin, proton-pump inhibitor, and poly-vitamins), they received tocilizumab intravenously in a dose of 8 mg/kg bodyweight (up to a maximum of 800 mg per dose), divided in two shots 12–24 h apart. Group B: Those received the standard care protocol alone, noting that guidelines were adjusted later on according to the updated scientific publications and WHO recommendations. The primary endpoint was to evaluate the effect of different regimens in controlling the disease, the need for mechanical ventilation and its duration (either invasive or non-invasive), length of ICU stay, hospital stay, and in-hospital mortality. Comparisons between quantitative variables were done using the non-parametric Mann–Whitney U-test. For comparison of serial measurements within each patient, the non-parametric Wilcoxon signed-rank test was used. For comparing categorical data, Chi-square (2) test was performed. Exact test was used instead when the expected frequency was <5. Correlations between quantitative variables were done using Spearman correlation coefficient. RESULTS: During this period, 166 patients were admitted to ICU, suffering from severe hypoxemia with moderate to severe COVID-19 pneumonia, 10 of them were excluded (three were over 80 years old, other three had advanced stages of malignancy, two were on steroids therapy and non-invasive home ventilation due to chronic chest condition, and two were presented with MODs and deceased in <48 h from admission), thus, 156 were included in the study. Group A: Seventy-six patients (49%) received tocilizumab in addition to standard therapy, Group B: Eighty patients (51%) received standard therapy only. In Group A, the mean length of ICU stay was 8.96 days with mean length of hospital stay 13.76, compared to mean length f ICU stay 9 days in Group B (p = 0.57) and mean length of hospital stay 12.46 days (p = 0.117). In Group A, 35 patients (46%) needed non-invasive mechanical ventilation (MV),12 patients of the 35 needed invasive MV in later stage, compared to 26 patients (32%) in Group B, 14 patients of the 26 needed invasive MV in later stage (p = 0.16). In Group A, 14 patients (18.4%) needed invasive mechanical ventilation, compared to 19 patients (23.7%) in Group B (p = 0.213). In Group A, 6 (7.9%) of 76 patients died, compared to 13 (16.3%) of 80 in Group B p = 0.11. The incidence of secondary bacterial infection in Group A was 16 patients (21%) compared to 21 (26%) in Group B (p = 0.44). CONCLUSION: In this study, we did not detect statistical difference in both groups of patients coming during CRS-associated COVID-19 pneumonia, regarding (ICU stay, need for and length of MV, the incidence of secondary bacterial infection, and in-hospital mortality) for COVID-19 moderate-to-severe pneumonia.

13.
European Journal of Clinical Investigation ; 52:160-161, 2022.
Article in English | EMBASE | ID: covidwho-1937924

ABSTRACT

Background: Familial Mediterranean fever (FMF) is an autosomal recessive autoinflammatory disease characterized by recurrent fever and serositis attacks. We aimed to explore clinical aspects of FMF including knowledge, diagnosis, symptoms, and medication in a Lebanese cohort enrolled by ad-hoc questionnaire. Methods: During November 2021-March 2022 we conducted an online survey by a google form questionnaire (33 items) advertised across Lebanese communities, hospitals, internists, and specialists. Patients and children's parents voluntarily provided information about FMF knowledge, diagnosis, presence, and severity of symptoms before and after medication. Since COVID-19 and FMF may share some common symptoms due to activation of the inflammasome pathway, we further investigated this aspect in the FMF cohort with symptomatic COVID-19. Results: A total of 123 FMF patients participated in this survey (75 females, age range 1-67 years;10 subjects from Armenia, Persia, and Turkey). The most frequent MEFV variants were M694V, M694I, E148Q, V726A, R202Q, and A744S. Before the diagnosis 70% of the subjects had no knowledge about FMF. The diagnosis was late in 40% of subjects (at age ≥20 years). A misdiagnosis occurred in 21% of subjects and was associated with unnecessary procedures such as heavy antibiotic prescription, appendectomy, and abdominal surgery. Prior to the diagnosis and targeted FMF therapy, subjects described typical febrile periodical attacks of systemic serositis with a frequency of more than attack once per month (48%) with intensity ranging from moderate to severe (95%). Following therapy with colchicine, 65% of the subjects reported mild symptoms. In addition, 60% of subjects had COVID-19 infection which was symptomatic in 80% of the cases. Concerning COVID-19, 63% of symptomatic COVID-19 subjects reported that FMF symptoms were higher compared to COVID-19, 23% reported that COVID-19 symptoms were higher than FMF symptoms, and 14% reported no difference between the two diseases. Additionally, 12% reported consequences of FMF-COVID- 19 combined symptoms, mainly joint pain due to persisting arthralgias. Conclusion: In Lebanon, an endemic region for FMF with a mixture of the ethnic communities from the Mediterranean area, FMF diagnosis can be missed, delayed, or initially erroneously classified. Nevertheless, the diagnostic ability is improving over time. This is the first study in Lebanon to clarify aspects of FMF knowledge, diagnosis, and symptoms as well as evaluation of COVID-19 and FMF interplay. The complex interaction and consequences between COVID-19 infection and the genetic autoinflammatory FMF is being further investigated.

14.
Journal of Hypertension ; 40:e29, 2022.
Article in English | EMBASE | ID: covidwho-1937690

ABSTRACT

Objective: COVID19-associated immunopathology is associated with increased production of interferon (IFN)-alpha (IFNα) and lambda3 (IFNL3). Effects of IFNs are mediated by interferon-stimulated genes (ISGs) and influence expression of angiotensin-converting enzyme 2 (ACE2), the receptor for S-protein (S1P) of SARS-CoV-2. Increasing evidence indicates vascular inflammation in cardiovascular sequelae of COVID19. We hypothesized that S1P-induced immune/inflammatory responses in endothelial cells (EC) are mediated via IFNα and IFNL3. Design and method: Human ECs were stimulated with S1P (1 μg/mL), IFNα (100ng/mL) or IFNL3 (100IU/mL). Because ACE2, metalloproteinase domain-17 (ADAM17) and type-II transmembrane serine protease (TMPRSS2) are important for SARS-CoV-2 infection, cells were treated with inhibitors of ADAM17 (marimastat, 3.8 nM), ACE2 (MLN4760, 440pM), and TMPRSS2 (camostat, 50 μM). Gene and protein expression was investigated by real-time PCR immunoblotting, respectively. Vascular function was assessed in mesenteric arteries from wild-type (WT) normotensive and hypertensive mice and in ISG15-deficient (ISG15KO) mice. Results: EC stimulated with S1P increased expression of IFNα (3-fold), IFNL3 (4-fold) and ISG (2-fold)(p < 0.05). EC exhibited higher responses to IFNα (ISG15: 16-fold) than to IFNL3 (ISG15: 1.7-fold)(p < 0.05). S1P increased gene expression of IL-6 (1.3-fold), TNFα (6.2-fold) and IL-1β (3.3-fold), effects that were maximized by IFNs. Only marimastat inhibited S1P effects. IL-6 was increased by IFNα (1,230pg/mL) and IFNL3 (1,124pg/mL) vs control (591pg/ mL). This was associated with increased phosphorylation of Stat1 (134%), Stat2 (102%), ERK1/2 (42%). Nitric oxide production and eNOS phosphorylation (Ser1177) were reduced by IFNα and (40%) and IFNL3 (40%). Reduced endothelium relaxation maximal response (%Emax) was observed in vessels from WTmice stimulated with IFNα (67%) and IFNL3 (71%) vs control (82%)(p < 0.05) but not in vessels from ISG15KO mice. Increased contraction was observed only in vessels from hypertensive mice treated with IFNα (9.1 ± 0.5mN vs control: 7.3 ± 0.3mN, p < 0.05). Conclusions: In ECs, S1P, IFNα and IFNL3 increased ISG15 and IL-6, processes that involve ADAM17. Inflammation induced by S1P was amplified by IFNs. IFNs induce vascular dysfunction through ISG15-dependent mechanisms, with augmented effects in hypertension. Our findings demonstrate that S1P induces immune/inflammatory responses that may be important in endotheliitis associated with COVID-19. This is especially important in the presence of cardiovascular risk factors, including hypertension.

15.
Handb. Exp. Pharmacol. ; 275:V-X, 2022.
Article in English | EMBASE | ID: covidwho-1929369
16.
American Journal of Respiratory and Critical Care Medicine ; 205(1), 2022.
Article in English | EMBASE | ID: covidwho-1927931

ABSTRACT

Rationale: The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has killed millions via the acute respiratory distress syndrome (ARDS). The early immune suppression of SARS-CoV-2 then subsequent inflammation suggests an unusual ability to cause immune dysregulation. Host transforming growth factor beta (TGF-β) is an immunesuppressing and profibrotic cytokine frequently “hijacked” by microbes to evade immune detection. We discovered a KRFK domain (a potent activating motif for latent TGF-β) in the SARS-CoV-2 nonstructural 15 (NSP15) protein. We hypothesized that this NSP15 protein causes immune dysregulation by activation of latent TGF-β and subsequent activation of immunosuppressive Tregulatory (Treg) cells, and that substantial TGF-β is present in the lungs of COVID-19 ARDS patients. Methods: We evaluated TGF-β1 concentrations in endotracheal aspirates (ETA) of 27 COVID-19 ARDS patients by ELISA. We produced recombinant SARS-CoV-2 NSP15 protein in E. coli and tested its ability to activate latent TGF-β1 using in vitro assays. TGF-β inhibitors were assessed for their ability to block effects. We obtained blood mononuclear cells from healthy subjects and isolated Tregs to assess their activation state via intracellular smad-2 phosphorylation (pSMAD2) using flow cytometry. Results: The KRFK domain was present in all SARS-CoV-2 variants. High concentrations of both active and total TGF-β1 were detected in ETA of COVID-19 ARDS patients (150 +/- 34 pg/ml active;1,819 +/- 304 pg/ml total) in a range previously shown to affect T cell function. NSP15 at 2.4 nM increased activation of latent TGF-β 12-fold (P < .001 vs. vehicle), compared to an 11% activation with the positive control thrombospondin-1 (TSP1;10 nM) (Figure). TGF-β receptor inhibitors blocked NSP15 effects on latent TGF-β activation and intracellular TGF-β1 signaling in a bioassay by over 95% (p<.01). At tested concentrations (25, 50, 100 nM) NSP15 increased Treg pSMAD2 levels via activation of latent TGF-β1, exceeding levels seen in Tregs stimulated with 400 pM of active TGF-β1 (+ control) (pSMAD2 + cells: vehicle 1.1%, active TGF-β1 43%, NSP15/latent TGF-β1 49-56%). Conclusions: High concentrations of active and total TGF-β1 are present in the lungs of COVID-19 ARDS patients, suggesting SARS-CoV-2 uses host TGF-β hijacking as a mechanism for immune evasion. The NSP15 protein of SARSCoV- 2 potently activates latent TGF-β, leading to Treg activation. TGF-β inhibitors are potent inhibitors of these NSP15 effects. A strategy to block NSP15-mediated effects with TGF-β inhibitors is an innovative therapy worthy of testing in animal models of COVID-19.

17.
American Journal of Respiratory and Critical Care Medicine ; 205(1), 2022.
Article in English | EMBASE | ID: covidwho-1927910

ABSTRACT

RATIONALE: The proteomic responses of hospitalized patients with SARS Co-V-2 infection may provide insight into risk, time course, and mechanisms associated with this infection. We used a high throughput proteomic platform to examine proteins that were differentially expressed relative to the length of hospital stay (LOS). METHOD:26 patients, hospitalized with SARS CoV-2 infection (mean age 48 yrs, 44% women) had blood samples obtained within 72 hours of admission. Initial plasma samples were analyzed from patients who were hospitalized for < 3 days (n=6), < 7days (n=12) and > 7 days (n=8) of LOS and compared to healthy controls (HC, n=8). Samples were analyzed with the modified aptamer-based array (SomaScan) that measures more than 7,000 human proteins representing different molecular pathways and gene families. Differentially regulated proteins with > 1.5 fold change and a false discovery rate of 5% were analyzed using the Ingenuity Pathway Analysis (IPA). Unique protein categories associated with LOS were assessed. RESULT: Compared to HC, differentially expressed proteins were detected among the 3 groups: 461 at < 3 days, 1,635 proteins at < 7 days and 1,738 proteins in >7 days. 407 proteins were common among all hospitalized COVID 19 individuals independent of LOS and 12, 250 and 361 proteins were uniquely present at < 3 days, < 7 days and > 7 days respectively compared to HC. The table below demonstrates the top highly enriched canonical pathway, molecular function and upstream regulator of differentially expressed proteins. The temporal sequence of these protein networks varied with LOS. Representative examples include early responses;platelet membrane glycoprotein GP6 signaling pathway that involves the FcR gamma-chain and the Src kinases linked to platelet aggregation, signaling involved in T cell receptor-mediated IL-2 production (TEC kinase). Less than 7 days include diacylglycerol associated with T cell activation, carnitine palmitoyltransferase associated with mitochondrial beta-oxidation of long chain fatty acids. CXCR4 a receptor for stromal -cell derived factor 1 and associated with COVID-19 prognosis. Late responses after 7 days include pathways involved in remodeling of epithelial adherens junctions. CONCLUSIONS : A high throughput proteomic approach provides insight into the dynamic regulation of protein pathways associated with the progression of SARS-Co-V2 infection. This may provide additional insight into risk and mechanisms associated with outcomes in COVID. (Table Presented).

18.
American Journal of Respiratory and Critical Care Medicine ; 205(1), 2022.
Article in English | EMBASE | ID: covidwho-1927905

ABSTRACT

Disorders in pulmonary vascular integrity are a prominent feature in many lung diseases, including acute respiratory distress syndrome (ARDS), capillary leak syndrome, and COVID19. Paracrine signals are enriched in the lung and are critically important in regulating the homeostasis of the functional pulmonary microvasculature. Here, we employed single-cell RNA-sequencing (scRNAseq) to study ligand and receptor interactions in the native human lung microvascular niche, and identified soluble factors that are critical in endothelial integrity. The scRNAseq data reveals a total of 47 cell populations consisting of five vascular endothelial subtypes in human lungs, including general capillary EC, aerocyte capillary EC (EC aCap), arterial EC, pulmonary venous EC, and systemic venous EC. Using EC aCap as a signal receiving core (Receptors) and the putative adjacent cell types (alveolar fibroblast, ATI, ATII, pericyte, plasma cell, etc.) in the EC aCap niche as senders (Ligands), we identified that SLIT2-ROBO4, ANGPT1-TIE1, ADM-RAMP2, VEGFD-KDR, and BMP5-BMPR2 are the top specific and abundant pairs in the niche. Immunostaining and ELISA assays confirmed their spatial information and secretion level. Furthermore, upon treatment with these ligands, real-time resistance recorded using an electric cell-substrate impedance sensing (ECIS) system revealed that VEGFD, ANGPT1 (angiopoietin 1), and ADM (adrenomedullin) could markedly increase the electrical resistance of human lung microvascular, arterial, and venous endothelial cells, suggesting their strong impact on the endothelial barrier function. Deciphering the cell-cell soluble signals that improve endothelial integrity in human lungs lays the foundation for uncovering the pathogenesis of pulmonary vascular disorders and the development of ex vivo functional lung vasculature.

19.
American Journal of Respiratory and Critical Care Medicine ; 205(1), 2022.
Article in English | EMBASE | ID: covidwho-1927902

ABSTRACT

Introduction: Dexamethasone decreases mortality in patients with severe COVID-19. The effects of dexamethasone on inflammation and repair in patients with severe COVID-19 are not well understood. We integrated tracheal aspirate (TA) and peripheral blood bulk/single-cell RNA sequencing to study the effect of dexamethasone on patients with COVID-19 ARDS. Methods: We studied selected patients from a cohort of adults with COVID-19 admitted to three hospitals in San Francisco, California from April 2020 to February 2021. Immunosuppression was not used to treat COVID-19 ARDS at these hospitals prior to July 2020, but was routinely used in these patients after this date. For this analysis, we included patients who were mechanically ventilated for COVID-19 ARDS for whom sequencing samples were available within four days of intubation. We excluded patients who received steroids prior to July 2020, subjects who received immunosuppression other than dexamethasone (e.g., tocilizumab) prior to sample collection, and chronically immunosuppressed subjects. We compared bulk RNASeq from TA and single cell RNASeq from TA and whole blood from subjects who received dexamethasone to subjects who did not receive dexamethasone. In addition, we studied the effect of dexamethasone on peripheral blood cytokine concentrations to confirm the effects of observed changes in gene expression. Results: TA bulk RNASeq was available from 20 subjects (six dexamethasone, 14 non-dexamethasone). There was no significant difference in age, sex, smoking, or BMI between groups. After correcting for multiple comparisons, 947 genes were differentially expressed in TA from subjects who received dexamethasone. Ingenuity Pathway Analysis predicted decreased activation of interferon, JAK/STAT, and NLRP12 signaling in subjects who received dexamethasone (Figure 1A). TA scRNASeq samples were available from ten dexamethasone-treated subjects and nine non-dexamethasone subjects. Whole blood scRNAseq samples were available for seven dexamethasone and eight non-dexamethasone subjects (Figure 1B). Eight subjects (three treated with dexamethasone) had both TA and whole blood scRNAseq samples available for analysis. Dexamethasone had distinct effects on the proportions of immune cells in tracheal aspirates and whole blood (Figure 1C). In 36 dexamethasone vs 42 non-dexamethasone subjects, treatment with dexamethasone was associated with significantly increased concentrations of IL-10 and decreased concentrations of IL-6 (Figure 1D). Conclusions: Dexamethasone decreases pro-inflammatory gene expression in the respiratory tract and peripheral blood of patients with COVID-19 ARDS. The effect of dexamethasone on specific cell populations may be distinct in the respiratory tract and peripheral blood.

20.
American Journal of Respiratory and Critical Care Medicine ; 205(1), 2022.
Article in English | EMBASE | ID: covidwho-1927857

ABSTRACT

Background: Latent class analyses in patients with acute respiratory distress syndrome (ARDS) have identified “hyper-inflammatory” and “hypo-inflammatory” phenotypes with divergent clinical outcomes and treatment responses. ARDS phenotypes are defined using plasma biomarkers and clinical variables. It is currently unknown if these phenotypes have distinct pulmonary biology and if pre-clinical models of disease replicate the biology of either phenotype. Methods: 45 subjects with ARDS (Berlin Definition) and 5 mechanically ventilated controls were selected from cohorts of mechanically ventilated patients at UCSF and ZSFG. Patients with COVID-19 were excluded from this analysis. A 3-variable classifier model (plasma IL-8, protein C, and bicarbonate;Sinha 2020) was used to assign ARDS phenotypes. Tracheal aspirate (TA) RNA was analyzed using established bulk and single-cell sequencing pipelines (Langelier 2018, Sarma 2021). Differentially expressed (DE) genes were analyzed using Ingenuity Pathway Analysis (IPA). Microbial community composition was analyzed with vegan. Fgsea was used to test for enrichment of gene sets from experimental ARDS models in genes that were differentially expressed between each phenotype and mechanically ventilated controls. Results: Bulk RNA sequencing (RNAseq) was available from 29 subjects with hypoinflammatory ARDS and 10 subjects with hyperinflammatory ARDS. 2,777 genes were differentially expressed between ARDS phenotypes. IPA identified several candidate upstream regulators of gene expression in hyperinflammatory ARDS including IL6, TNF, IL17C, and interferons (Figure 1A). 2,953 genes were differentially expressed between hyperinflammatory ARDS and 5 ventilated controls;in contrast, only 243 genes were differentially expressed between hypoinflammatory ARDS and controls, suggesting gene expression in the hypoinflammatory phenotype was more heterogeneous. Gene sets from experimental models of acute lung injury were enriched in hyperinflammatory ARDS but not in hypoinflammatory ARDS (Figure 1B). Single cell RNA sequencing (scRNAseq) was available from 6 additional subjects with ARDS, of whom 3 had hyperinflammatory ARDS. 14,843 cells passed quality control filters. Hyperinflammatory ARDS subjects had a markedly higher burden of neutrophils (Figure 1C), including a cluster of stressed neutrophils expressing heat shock protein RNA that was not present in hypoinflammatory ARDS. Expression of a Th1 signature was higher in T cells from hyperinflammatory ARDS. Differential expression analysis in macrophages identified increased expression of genes associated with mortality in a previous study of ARDS patients (Morell 2019). Conclusions: The respiratory tract biology of ARDS phenotypes is distinct. Hyperinflammatory ARDS is characterized by neutrophilic inflammation with distinct immune cell polarization. Transcriptomic profiling identifies candidate preclinical disease models that replicate gene expression observed in hyperinflammatory ARDS.

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