RETROSPECTIVE STUDY OF VITAMIN D DEFICIENCY IN RHEUMATOLOGIC PATIENTS DURING THE COVID-19 PANDEMIC IN POLAND

BackgroundBefore the COVID-19 pandemic it was estimated that nearly 70% of the population is deficient in vitamin D - 25(OH)D <20ng/ml in Poland [1]. The percentage was expected to increase due to indoor isolation during the COVID-19 pandemic. Vitamin D has a positive effect on the condition of the bones, affects the course of autoimmune diseases, the course of neurological diseases, in type 2 diabetes, vitamin D supplementation improves glucose tolerance and reduces insulin resistance [2,3,4].ObjectivesThe aim of the retrospective study was to determine what percentage of rheumatology clinic patients suffer from vitamin D deficiency and whether this condition is effectively treated.MethodsIn January 2023, a retrospective analysis of the documentation of 172 patients treated at the Rheumatology Outpatient Clinic in Bełżyce (Poland) in 2022 was conducted.ResultsResults: The mean age of the 172 patients whose documentation was analyzed was 60.43 years (min 19, max 88). There were 132 women (76.8%) and 40 men (23.2%) in this group. The mean concentration of vitamin D was 25.57ng/ml±SD11.9 (min 5.7, max 75, Me 22.8). Vitamin D deficiency was found in 44% (serum concentration <20mg/ml), suboptimal concentration (20-30ng/ml) in 31%, optimal concentration (30-50ng/ml) in 21%, and high concentration (>50ng/ml) ml) in 4%. All those with a deficit or deficiency (75 people) were prescribed cholecalciferol in a dose of 20,000 units orally, 1 capsule twice a week after breakfast for 2 months [5]. Patients with optimal vitamin D levels were advised to take a dose of 2,000 units per day. Among the patients with deficit or deficiency, 48 people came for a follow-up visit to check the level of vitamin D (64% of the group with too low vitamin D concentration;28% of the entire group whose documentation was analyzed). In the follow-up examination, the mean concentration of vitamin D was 37.14±9.8ng/ml (min 28, max 84, Me 35.3). Therefore, a statistically significant increase in the concentration of vitamin D in the blood was noted (p<0.05). In the group of people who came for the follow-up examination, there were 35 women, whose mean age was 60.7 years and 13 men (mean age 68.2 years).Conclusion:1. During the COVID-19 pandemic in the group of outpatient rheumatology patients, 75% had a deficiency or suboptimal level of vitamin D.2. Treatment with cholecalciferol in a dose of 20,000 IU twice a week orally for 2 months is effective treatment of vitamin D deficiency.3. Too low percentage of patients diagnosed with vitamin D deficiency come for visits and check-ups.References[1]Hilger J., Friedel A., Herr R.. A systematic review of vitamin D status in populations worldwide. Br J Nutr. 2013;9: 1023.[2]Karczmarewicz E., Czekuć-Kryskiewicz E., Płudowski P. Effect of vitamin D status on pharmacological treatment efficiency-impact on cost- effective management in medicine. Dermatoendocrinology, 2013;5: 299-304.[3]Zhu J., Bing C., Wilding J.P.H. Vitamin d receptor ligands attenuate the inflammatory profile of IL-1β-stimulated human white preadipocytes via modulating the NF-κB and unfolded protein response pathways Biochemical and Biophysical Research Communications 2-18, 503: 1049-1056.[4]Luan W., Hammond L.A. Vuillermot S. Maternal vitamin d prevents abnormal dopaminergic development and function in a mouse model of prenatal immune activation. Scientific Reports 2018;8 (1) article numer 9741.[5]Płudowski P., Karczmarewicz E. i wsp. Witamina D: Rekomendacje dawkowania w populacji osób zdrowych oraz w grupach ryzyka deficytów.Wytyczne dla Europy Środkowej 2013 r. Standardy Medyczne/Pediatria 2013, 10, 573-578 (in Polish).Acknowledgements:NIL.Disclosure of InterestsNone Declared.

Repurposing of known drugs for COVID-19 using molecular docking and simulation analysis

We selected fifty one drugs already known for their potential disease treatment roles in various studies and subjected to docking and molecular docking simulation (MDS) analyses. Five of them showed promising features that are discussed and suggested as potential candidates for repurposing for COVID-19. These top five compounds were boswellic acid, pimecrolimus, GYY-4137, BMS-345541 and triamcinolone hexacetonide that interacted with the chosen receptors 1R42, 4G3D, 6VW1, 6VXX and 7MEQ, respectively with binding energies of -9.2 kcal/mol, -9.1 kcal/mol, -10.3 kcal/mol, -10.1 kcal/mol and -8.7 kcal/mol, respectively. The MDS studies for the top 5 best complexes revealed binding features for the chosen receptor, human NF-kappa B transcription factor as an important drug target in COVID-19-based drug development strategies.

Signaling pathways implicated in SARS-CoV-2 infection: key drug targets for human coronaviruses and related viral infections

The occurrence of SARS-CoV-2 in 2019 is the second coronavirus spread-out after the SARS-CoV, which has pandemic potential. Search for its remedies is dependent on integrative knowledge of cell signaling pathways, which is under clinical scrutiny. The major cascades triggered by coronavirus entry include Renin-Angiotensin System, MAPK, NF-κB, JAK/STAT which are involved with innate immunity. Some other modes are, unfolded protein response signaling and inflammasome mediated apoptosis activation. Virulence factors of the SARS-CoVs like spike, envelope, nonstructural proteins etc., interfere with some of these viral defense pathways. Therapeutically, the viral intrusion, multiplication, as well as tissue-injurious cytokine overreactions are targeted by pathway-specific drugs. Viral entry blockers, p38 MAPK inhibitors, cytokine regulators, JAK inhibitors, and anti-inflammatory drugs are either being repurposed or innovated with scopes for futuristic modeling. This chapter is aimed to elucidate the pathological signaling network behind Severe Acute Respiratory Syndrome, for evaluation of existing and postulated drug targets. © 2022 Elsevier Inc. All rights reserved.

Immunomodulatory effects of Abelmoschus esculentus water extract through MAPK and NF-κB signaling in RAW 264.7 cells

During the current COVID-19 pandemic, the world is facing a new, highly contagious virus that suppresses innate immunity as one of its early virulence mechanisms. Therefore, finding new methods to enhance innate immunity is a promising strategy to attenuate the effects of this major global health problem. With the aim of characterizing bioactive ingredients as immune-enhancing agents, this study focuses on Abelmoschus esculentus (okra), which has several previously demonstrated bioactivities. Firstly, we investigated the immune-stimulatory effects of okra leaf ethanol extract (OLE) and okra leaf water extract (OLW) on nitric oxide (NO) production in macrophages. OLE significantly decreased nitrite accumulation in LPS-stimulated RAW 264.7 cells, indicating that it potentially inhibited NO production in a concentration-dependent manner. In contrast, OLW significantly enhanced the production of prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and NO in a dose-dependent manner. OLW also increased the expression levels of NO synthase (iNOS) and cyclooxygenase (COX)-2, potentially explaining the OLW-induced increase in NO and PGE2 production. In addition, OLW stimulated the phosphorylation of mitogen-activated protein kinases (MAPKs;ERK, p38, and JNK) as well as the activation and subsequent nuclear translocation of nuclear factor κB (NF-κB). This indicated that OLW activates macrophages to secrete PGE2, TNF-α, IL-1β, and NO, inducing iNOS and COX-2 expression via activation of the NF-κB and MAPK signaling pathways. In conclusion, our results demonstrate that OLW can effectively promote the activation of macrophages, suggesting that OLW may possess potent immunomodulatory effects and should be explored as a potential health-promoting materials to boost the immune system.

Computational drug repurposing against SARS-CoV-2 reveals plasma membrane cholesterol depletion as key factor of antiviral drug activity

Comparing SARS-CoV-2 infection-induced gene expression signatures to drug treatment-induced gene expression signatures is a promising bioinformatic tool to repurpose existing drugs against SARS-CoV-2. The general hypothesis of signature-based drug repurposing is that drugs with inverse similarity to a disease signature can reverse disease phenotype and thus be effective against it. However, in the case of viral infection diseases, like SARS-CoV-2, infected cells also activate adaptive, antiviral pathways, so that the relationship between effective drug and disease signature can be more ambiguous. To address this question, we analysed gene expression data from in vitro SARS-CoV-2 infected cell lines, and gene expression signatures of drugs showing anti-SARS-CoV-2 activity. Our extensive functional genomic analysis showed that both infection and treatment with in vitro effective drugs leads to activation of antiviral pathways like NFkB and JAK-STAT. Based on the similarity—and not inverse similarity—between drug and infection-induced gene expression signatures, we were able to predict the in vitro antiviral activity of drugs. We also identified SREBF1/2, key regulators of lipid metabolising enzymes, as the most activated transcription factors by several in vitro effective antiviral drugs. Using a fluorescently labeled cholesterol sensor, we showed that these drugs decrease the cholesterol levels of plasma-membrane. Supplementing drug-treated cells with cholesterol reversed the in vitro antiviral effect, suggesting the depleting plasma-membrane cholesterol plays a key role in virus inhibitory mechanism. Our results can help to more effectively repurpose approved drugs against SARS-CoV-2, and also highlights key mechanisms behind their antiviral effect.

Research progress on antiviral mechanism of artemisinin and its derivatives

Artemisinin, extracted from the traditional Chinese medicine Artemisia annua Linn., has a powerful antimalarial effect. With further research, artemisinin and its derivatives have been found to have antiviral effects against a variety of viruses from different viral families, including Herpesviridae, Flaviridae, and Coronaviraceae, and can inhibit virus-induced inflammatory responses. The possible antiviral mechanisms include alkylation of DNA binding domain of NF-κB P65 subunit, inhibition of plasmidic nuclear transport of NF-κB p65, production of reactive oxygen species or activation of carbon center free radicals to regulate host Keap1/Nrf2/ARE pathway to inhibit viral replication. The inflammatory response was alleviated by inhibiting the activation of NF-κB and ERK pathways induced by the virus. This paper reviews the possible antiviral mechanism of artemisinin and its derivatives to provide reference for artemisinin to fight new viruses and promote new use of old drugs. © 2022 by the authors.

Tissue Integrity and COVID-19

DefinitionTissue integrity depends on biological tissue stiffness. Tissue integrity can protect both against age-related diseases and against severity of COVID-19. The disruption of tight junctions and increase of tissue permeability with advancing age can be related with age-related diseases as well as with age-dependent COVID-19. Release of tightly bound water from collagen fibrils leads to the increase of extracellular matrix stiffness and to the associated with matrix stiffness increased tissue permeability. The link between arterial stiffness and oxidative stress has been reported and is expected to be studied in more detail in the future. Trehalose can be suggested for retardation of tightly bound water release and subsequent extracellular matrix crosslinking by advanced glycation end products. Increase in tissue permeability can be blocked by polyphenols that inhibit ICAM-1 expression and mitigate cytoskeleton reorganization. NF-κB activation as a result of increased stiffness and cytoskeleton reorganization can cause both cardiovascular pathologies and COVID-19. Increased cholesterol content in cell membrane leads to increased virus entry into cell and increase of cholesterol is linked with cardiovascular diseases. Statins and chitosan are known as cholesterol-lowering substances. Nrf2 inhibits NF-κB activation and NF-κB inhibits Nrf2 pathway.