International Journal of Research and Reports in Hematology

Assessment of Some Inflammatory Cytokines of Interest in Patients with Severe Acute Respiratory Syndrome Coronavirus 2 Admitted in Isolation Centres in Port-Harcourt

Chukwuka Achonnwo Kpaluku *

Department of Medical Laboratory Science, Rivers State University, Port Harcourt, Nigeria.

Evelyn M. Eze

Department of Medical Laboratory Science, Rivers State University, Port Harcourt, Nigeria.

Stella Urekweru Ken-Ezihuo

Department of Medical Laboratory Science, Rivers State University, Port Harcourt, Nigeria.

*Author to whom correspondence should be addressed.

---

Abstract

Coronavirus disease 2019 (COVID-19) is a novel and highly contagious viral infection caused by Sars-cov-2 and has been associated with a hyper-inflammatory immune response. This study aimed to evaluate changes in some inflammatory cytokines as clinical biomarkers to aid in the effective management of COVID-19 progression in Port Harcourt. A case-control study design was employed in this study, where a total of one hundred and ten (110) subjects were recruited, comprising fifty-five (55) COVID-19 positive subjects and fifty-five (55) COVID-19 negative subjects (controls), all between the ages of twenty (20) and seventy (70) years old, and both male and female subjects. Five milliliters (5 ml) of whole blood was collected using standard venipuncture technique with sterile hypodermic syringes and needles, aseptically, and dispensed into a plain bottle for the analysis of inflammatory cytokines. For the confirmation of COVID-19 positivity, a nasopharyngeal swab was collected aseptically using the RT-PCR technique. The results of this study revealed a significantly increased level of IL-6 (p = 0.0399) among subjects with COVID-19. The results from this study indicate significant alterations in inflammatory cytokines among subjects with COVID-19. It is necessary that tertiary health care settings and isolation centres consider IL-6 as a biomarker for effective management of patients with COVID-19 disease.

Keywords: COVID-19, cytokines, RT-PCR technique, pneumonia

---

---

---

References

World Health Organization (WHO). Coronavirus Disease (COVID-19) Situation Reports. Geneva Switzerland: World Health Organization; 2020a. Available:https://www.who.int/emergencies/diseases/ novel-coronavirus-2019/situation-reports

Accessed 5 July 2021.

Jayasinghe R, Ranasinghe S, Jayarajah U, Seneviratne S. Quality of online information for the general public on COVID-19. Patient Education and Counseling. 2020;103(1):2594-2597.

United Nations International Children's Emergency Fund. The evolving epidemiologic and clinical picture of SARS-CoV-2 and COVID-19 disease in children and young people. UNICEF Office of Research 2020; 2020.

Available:https://www.unicef-irc.org/publications/pdf/

Accessed on 5 July 2021.

Weiss SR, Leibowitz JL. Coronavirus pathogenesis. Advanced Virus Research. 2011;81(1):85-164.

Richman DD, Whitley RJ, Hayden FG. Pathogenesis of disease causation. Clinical virology 4th edn. Washington: ASM Press; 2016.

Yang D, Leibowitz JL. The structure and functions of coronavirus genomic 3’ and 5’ ends. Virus Research. 2015;206(1):120-133.

Zaki AM, Van-Boheemen S, Bestebroer TM, Osterhaus AD, Fouchier RA. Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. New England Journal Medicine. 2012;367(19):1814-1820.

Drosten C, Günther S, Preiser W. Identification of a novel coronavirus in patients with severe acute respiratory syndrome. New England Journal of Medicine. 2020;348 (1):1967-1976.

Cui J, Li F, Shi Z. Origin and evolution of pathogenic coronaviruses. Nature Reviews Microbiology. 2019;17(1):181-192.

Li Q, Guan X, Wu P. Early transmission dynamics in Wuhan China of novel coronavirus-infected pneumonia. New England Journal of Medicine. 2020a;3(5):21-30.

Li J, Yuan P, Heffernan J, Zheng T, Ogden N, Sander B, Zhu H. (Fangcang shelter hospitals during the COVID-19 epidemic Wuhan China. Bulletin of the World Health Organization. 2020b;98(12):830-841.

Beaney T, Neves A. L, Alboksmaty A, Ashrafian H, Flott K, Fowler A, Clarke J. Trends and associated factors for COVID-19 hospitalisation and fatality risk in 2.3 million adults in England. Nature Communications. 2022;13(1):1-10.

Li F. Structure function and evolution of coronavirus spike proteins. Annual Review of Virology. 2016;3 (1):237-261.

Cucinotta D, Vanelli M. WHO declares COVID-19 a pandemic. Acta Bio Medical Atenei Parmensis. 2020;91(1):157 - 182.

Woo PC, Lau SK, Huang Y, Yuen KY. Coronavirus diversity phylogeny and interspecies jumping. Experimental Biology and Medicine. 2019;234(10):1117-1127.

Jayasekara D, Seneviratne SL, Jayasekara A, De Zoysa I. Atypical presentations of COVID-19. Advanced Infectious Diseases. 2020;10 (1):136- 142.

Chan JF, Lau SK, To KK, Cheng VC, Woo PC, Yuen KY. Middle East respiratory syndrome coronavirus: another zoonotic betacoronavirus causing SARSlike disease. Clinical Microbiology Reviews. 2015;28(2):465-522.

Hui DS. An overview on severe acute respiratory syndrome (SARS). Monaldi Archives for Chest Disease. 2005;63(3):149-157.

Pal M, Berhanu G, Desalegn C, Kandi V. Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2): An update. Cureus. 2020;12(3):8721 - 8729.

Rahman A, Niloofa R, Jayarajah U, De Mel S, Abeysuriya V, Seneviratne SL. Hematological abnormalities in COVID-19: A narrative review. The American Journal of Tropical Medicine and Hygiene. 2021;104(4):1188-1194.

Centers for disease control and prevention. First travel-related case of 2019 novel coronavirus detected in United States; 2020. Available:https://www.cdc.gov/media/releases/2020/p0121-novel-coronavirus-travel-case.html (accessed July 18 2020).

Rahbari R, Moradi N, Abdi M. rRT-PCR for SARS-CoV-2: Analytical considerations. Clinica Chimica Acta. 2021;516 1-7.

D'Cruz RJ, Currier AW, Sampson VB. Laboratory testing methods for novel severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2). Frontiers in Cell and Developmental Biology. 2020;8:468-474.

Wu C, Chen X, Cai Y, Xia J, Zhou X, Xu S, Huang H, Zhang L, Zhou X, Du C, Zhang Y, Song J, Wang S, Chao Y, Yang Z, Xu J, Zhou X, Chen D, Xiong W, Xu L, Song Y. Risk Factors associated with acute respiratory distress syndrome and death in patients with coronavirus disease 2019 Pneumonia in Wuhan China. Journal of American Medical Association of Internal Medicine. 2020;180(7):934-943.

Jin YH, Cai L, Cheng ZS, Cheng H, Deng T, Fan YP, Wang XH. A rapid advice guideline for the diagnosis and treatment of 2019 novel coronavirus (2019-nCoV) infected pneumonia (standard version). Military Medical Research. 2020;7(1):1-23.

Stoecklin SB, Rolland P, Silue Y, Mailles A. First cases of coronavirus disease 2019 (COVID-19) in France: Surveillance investigations and control measures 2020. Euro Surveillance. 2020;25 (6):1-10.

World Health Organization (WHO). Coronavirus Disease (COVID-19) Technical Guidance: Laboratory Testing for 2019-nCoV in Humans; 2020b. Available:https://www.who.int/emergencies/diseases/novel-coronavirus-2019/technical~guidance/laboratory-guidance

(accessed 10 July 2021)

Kumar S, Nyodu R, Maurya VK, Saxena SK. Morphology genome organization replication and pathogenesis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In Coronavirus Disease 2019 (COVID-19). Springer Singapore. 2020;23-31.

Andersen KG, Rambaut A, Lipkin WI. The proximal origin of SARS-CoV-2. Nature Medicine. 2020;26 (4):450-452.

Shereen MA, Khan S, Kazmi A, Bashir N, Siddijque R. COVID-19 infection: Origin transmission and characteristics of human coronaviruses. Journal of Advanced Research. 2020;24(1):91-98.

Park WB, Kwon NJ, Choi SJ, Kang CK, Choe PG, Kim JY, Yun J, Lee GW, Seong MW, Kim NJ, Seo JS, Oh MD. Virus isolation from the first patient with SARS-CoV-2 in Korea. Journal of Korean Medical Science. 2020;35(7):84-90.

Finlay BB, See RH, Brunham RC. Rapid response research to emerging infectious diseases: Lessons from SARS. Nature Reviews Microbiology. 2004;2(7):602-607.

Saxena S. K, Kumar S, Maurya VK, Sharma R, Dandu HR, Bhatt M. Current Insight into the novel coronavirus disease 2019 (COVID-19). Coronavirus Disease 2019 (COVID-19). Epidemiology Pathogenesis Diagnosis and Therapeutics. 2020;1(1):1-8.

Luo M, Guo L, Yu M, Jiang W, Wang H. The psychological and mental impact of coronavirus disease 2019 (COVID-19) on medical staff and general public–A systematic review and meta-analysis. Psychiatry Research. 2020;291:1131-1139.

Walls AC, Park YJ, Tortorici MA, Wall A, McGuire AT, Veesler D. Structure function and antigenicity of the SARS-CoV-2 spike glycoprotein. Cell. 2020;181(2):281-292.

Coutard B, Valle C, de Lamballerie X, Canard B, Seidah NG, Decroly E. The spike glycoprotein of the new coronavirus 2019-nCoV contains a furin-like cleavage site absent in CoV of the same clade. Antiviral Research. 2020;176 (1):104-110.

Kamimura H, Kamimura K, Tsuchiya A, Terai S. Successful treatment of positive-sense RNA virus coinfection with autoimmune hepatitis using double filtration plasmapheresis. British Medical Journal of Case Reports. 2021;14(3):236984-236987.

Samprathi M, Jayashree M. Biomarkers in COVID-19: An up-to-date review. Frontiers in Pediatrics. 2021;1(8):1- 12.

Hoffmann M, Kleine-Weber H, Schroeder S, Krüger N, Herrler T, Erichsen S, Schiergens TS, Herrler G, Wu N. H, Nitsche A, Müller MA, Drosten C, Pöhlmann S. SARS-CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell. 2020;181 (1):1-10.

Agarwal A, Chen A, Ravindran N, To C, Thuluvath PJ. Gastrointestinal and liver manifestations of COVID-19. Journal of Clinical and Experimental Hepatology. 2020;10(3):263-265.

Wrapp D, Wang N, Corbett KS, Goldsmith JA, Hsieh CL, Abiona O, Graham BS, McLellan JS. Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation. Science. 2020;367 (6483):1260-1263.

Rabaan AA, Mutair AA, Alawi ZA, Alhumaid S, Mohaini MA, Aldali J, Dhama K. Comparative pathology molecular pathogenicity immunological features and genetic characterization of three highly pathogenic human coronaviruses (MERS-CoV SARS-CoV and SARS-CoV-2). European Review of Medical and Pharmacological Science. 2021;25(22): 7162-7184.

Zhang L, Dermawan K, Jin M, Liu R, Zheng H, Xu L, Xiong S. Differential impairment of regulatory T cells rather than effector T cells by paclitaxel-based chemotherapy. Clinical Immunology. 2008;129(2):219-229.

Averyanov A, Kogan E, Lesnyak V, Danilevskaya O. Lung disease related to connective tissue diseases. In Difficult to Diagnose Rare Diffuse Lung Disease 265-319. Academic Press; 2020.

Xu Z, Shi L, Wang Y, Zhang J, Huang L, Zhang C, Liu S, Zhao P, Liu H, Zhu L, Tai Y, Bai C, Gao T, Song J, Xia P, Dong J, Zhao J, Wang FS. Pathological findings of COVID-19 associated with acute respiratory distress syndrome. Lancet Respiratory Medicine. 2020;8 (4):420-422.

Tian S, Hu W, Niu L, Liu H, Xu H, Xiao SY. Pulmonary pathology of early-phase 2019 novel coronavirus (COVID-19) pneumonia in two patients with lung cancer. Journal of Thoracic Oncology: Official Publication of the International Association for the Study of Lung Cancer. 2020;15 (5):700-704.

Huang C, Wang Y, Li X, Ren L, Zhao J, Hu Y, Cao B. Clinical features of patients infected with 2019 novel coronavirus in Wuhan China. The Lancet. 2020;395(10223):497-506.

Wang L, Wang Y, Ye D, Liu Q. Review of the 2019 novel coronavirus (SARS-CoV-2) based on current evidence. International Journal of Antimicrobial Agents. 2020;55(6):105948.

Fuk-Woo CJ, Shuofeng Y, Kin-Hang K. A familial cluster of pneumonia associated with the 2019 novel coronavirus indicating person-to-person transmission: A study of a family cluster. Lancet. 2020;395(1):514-523.

Shi Y, Wang Y, Shao C, Huang J, Gan J, Huang X, Bucci E, Piacentini M, Ippolito G, Melino G. COVID-19 infection: The perspectives on immune responses. Cell Death and Differentiation. 2020;27 (5):1451-1454.

Osorio L, Caraglia M, Facchini G, Margherita V, Placido SD, Buonerba C. Toll-like receptors and COVID-19: A two-faced story with an exciting ending. Future Science OA. 2020;6(8):605-609.

Conti P, Ronconi G, Craffa A, Gallenga CE, Ross R, Frydas I. Induction of pro-inflammatory cytokines (IL-1 and IL-6) and inflammation by COVID-19: Anti-inflammatory strategies. Journal of Biological Regulators and Homeostatic Agents. 2020;34 (2):327-331.

Chen X, Zhao B, Qu Y, Chen Y, Xiong J, Feng Y, Li F. Detectable serum SARS-CoV-2 viral load (RNAaemia) is closely correlated with drastically elevated interleukin 6 (IL-6) level in critically ill COVID-19 patients. Clinical Infectious Diseases. 2020;8(4):2188 - 2194.

Mardi A, Meidaninikjeh S, Nikfarjam S, Majidi Zolbanin N, Jafari R. Interleukin-1 in COVID-19 infection: Immunopathogenesis and possible therapeutic perspective. Viral Immunology. 2021; 34(10):679-688.

Newton K, Dixit VM. Signaling in innate immunity and inflammation. Cold Spring Harbor Perspectives in Biology. 2012;4(3):604-609.

Ma J, Chen T, Mandelin J, Ceponis A, Miller N. E, Hukkanen M, Konttinen YT. Regulation of macrophage activation. Cellular and Molecular Life Sciences CMLS. 2003;60(11):2334-2346.

Netea MG, Stuyt RJ, Kim SH, Van der Meer JW, Kullberg BJ, Dinarello CA. The Role of Endogenous Interleukin (IL)--18 IL-12 IL-1b and Tumor Necrosis Factor--a in the Production of Interferon-g Induced by Candida albicans in Human Whole-Blood Cultures. Journal of Infectious Diseases. 2002;185(7):871 - 879.

Dinarello CA. The IL-1 family and inflammatory diseases. Clinical and Experimental Rheumatology. 2002;20(5):1-13.

Krishnaswamy JK, Chu T, Eisenbarth SC. Beyond pattern recognition: NOD-like receptors in dendritic cells. Trends in Immunology. 2013;34(5):224-233.

Yaqinuddin A, Kashir J. Novel therapeutic targets for SARS-CoV-2-induced acute lung injury: Targeting a potential IL-1β/neutrophil extracellular traps feedback loop. Medical Hypotheses. 2020;143 1096-1098.

Costela-Ruiz VJ, Illescas-Montes R, Puerta-Puerta JM, Ruiz C, Melguizo-Rodríguez L. SARS-CoV-2 infection: The role of cytokines in COVID-19 disease. Cytokine & Growth Factor Reviews. 2020;54 62-75.

Toniato E, Ross R, Kritas SK. How to reduce the likelihood of coronavirus-19 (CoV-19 or SARS-CoV-2) infection and lung inflammation mediated by IL-1. Journal of Biological Regulators and Homeostatic Agents. 2020;34(2):11-16.

Divani AA, Andalib S, Di Napoli M, Lattanzi S, Hussain M. S, Biller J, Torbey M. Coronavirus disease 2019 and stroke: Clinical manifestations and pathophysiological insights. Journal of Stroke and Cerebrovascular Diseases. 2020;29(8):104941-104948.

Ceribelli A, Motta F, De Santis M, Ansari AA, Ridgway WM, Gershwin ME, Selmi C. Recommendations for coronavirus infection in rheumatic diseases treated with biologic therapy. Journal of Autoimmunity. 2020;109:1024-1029.

Duong P, Tenkorang MA, Trieu J, McCuiston C, Rybalchenko N, Cunningham RL. Neuroprotective and neurotoxic outcomes of androgens and estrogens in an oxidative stress environment. Biology of Sex Differences. 2020;11(1):1-18.

Batu ED, Özen S. Implications of COVID-19 in pediatric rheumatology. Rheumatology International. 2020;40(8): 1193-1213.

Manji H, Carr AS, Brownlee WJ, Lunn MP. Neurology in the time of COVID-19. Journal of Neurology Neurosurgery & Psychiatry. 2020;91(6):568-570.

Jamilloux Y, Henry T, Belot A, Viel S, Fauter M, El Jammal T, Sève P. Should we stimulate or suppress immune responses in COVID-19? Cytokine and anti-cytokine interventions. Autoimmunity Reviews. 2020;19(7):1025-1033.

Gautret P, Million M, Jarrot PA, Camoin-Jau L, Colson P, Fenollar F, Raoult D. Natural history of COVID-19 and therapeutic options. Expert Review of Clinical Immunology. 2020; 16(12):1159-1184.

Kishimoto T. Interleukin-6: From basic science to medicine-40 years in immunology. Annual Review of Immunology. 2005;23(1):1-21.

Potere N, Batticciotto A, Vecchié A, Porreca E, Cappelli A, Abbate A, Bonaventura A. The role of IL-6 and IL-6 blockade in COVID-19. Expert Review of Clinical Immunology. 2021; 17(6):601-618.

Kang S, Narazaki M, Metwally H, Kishimoto T. Historical overview of the interleukin-6 family cytokine. Journal of Experimental Medicine. 2020;217(5):231 - 238.

Copaescu A, Smibert O, Gibson A, Phillips EJ, Trubiano JA. The role of IL-6 and other mediators in the cytokine storm associated with SARS-CoV-2 infection. Journal of Allergy and Clinical Immunology. 2020;146(3):518-534.

Cavalli G, Farina N, Campochiaro C, De Luca G, Della-Torre E, Tomelleri A, Dagna L. Repurposing of biologic and targeted synthetic anti-rheumatic drugs in COVID-19 and hyper-inflammation: A comprehensive review of available and emerging evidence at the peak of the pandemic. Frontiers in Pharmacology. 2020;11:598308.

Horby P. W, Pessoa-Amorim G, Peto L, Brightling CE, Sarkar R, Thomas K, Landray MJ. Tocilizumab in patients admitted to hospital with COVID-19 (RECOVERY): Preliminary results of a randomised controlled open-label platform trial. Medrxiv. 2021;5(2):213 - 219.

Siddiqi HK, Mehra MR. COVID-19 illness in native and immunosuppressed states: A clinical–therapeutic staging proposal. The Journal of Heart and Lung Transplantation. 2020;39(5):405-407.

Polidoro RB, Hagan RS, de Santis Santiago R, Schmidt NW. Overview: systemic inflammatory response derived from lung injury caused by SARS-CoV-2 infection explains severe outcomes in COVID-19. Frontiers in Immunology. 2020;11:1626.

Herold T, Jurinovic V, Arnreich C, Lipworth BJ, Hellmuth JC, von Bergwelt-Baildon M, Weinberger T. Elevated levels of IL-6 and CRP predict the need for mechanical ventilation in COVID-19. Journal of Allergy and Clinical Immunology. 2020;146(1):128-136.

Bost P, Giladi A, Liu Y, Bendjelal Y, Xu G, David E, Amit I. Host-viral infection maps reveal signatures of severe COVID-19 patients. Cell. 2020;181(7):1475-1488.

Diao B, Wang C, Tan Y, Chen X, Liu Y, Ning L, Chen Y. Reduction and functional exhaustion of T cells in patients with coronavirus disease 2019 (COVID-19). Frontiers in Immunology. 2020; 827:2121 - 2129.

Ruan Q, Yang K, Wang W, Jiang L, Song J. Clinical predictors of mortality due to COVID-19 based on an analysis of data of 150 patients from Wuhan China. Intensive Care Medicine. 2020;46(1):846-848.

McFadyen JD, Stevens H, Peter K. The emerging threat of (micro) thrombosis in COVID-19 and its therapeutic implications. Circulation Research. 2020;127(4):571-587.

Grant RA, Morales-Nebreda L, Markov NS, Swaminathan S, Querrey M, Guzman ER, Wunderink RG. Circuits between infected macrophages and T cells in SARS-CoV-2 pneumonia. Nature. 2021;590(7847):635-641.

Patra T, Meyer K, Geerling L, Isbell TS, Hoft DF, Brien J, Ray R. SARS-CoV-2 spike protein promotes IL-6 trans-signaling by activation of angiotensin II receptor signaling in epithelial cells. Public Library of Science Pathogens. 2020;16(12): e1009128.

Gubernatorova EO, Polinova AI, Petropavlovskiy MM, Namakanova OA, Medvedovskaya AD, Zvartsev RV, Nedospasov SA. Dual role of TNF and LTα in carcinogenesis as implicated by studies in mice. Cancers. 2021;13(8):1775 - 1779.

Canzano P, Brambilla M, Porro B, Cosentino N, Tortorici E, Vicini S, Camera M. Platelet and endothelial activation as potential mechanisms behind the thrombotic complications of COVID-19 patients. Basic to Translational Science. 2021;6(3):202-218.

Atzeni F, Talotta R, Salaffi F, Cassinotti A, Varisco V, Battellino M, Sarzi-Puttini P. Immunogenicity and autoimmunity during anti-TNF therapy. Autoimmunity Reviews. 2013; 12(7):703-708.

Ablamunits V, Lepsy C. Blocking TNF signaling may save lives in COVID-19 infection. Molecular Biology Reports. 2022;1-7.

Malaviya R, Laskin JD, Laskin DL. Anti-TNFα therapy in inflammatory lung diseases. Pharmacology & Therapeutics. 2017;180 90-98.

Yao Y, Zhou J, Diao X, Wang S. Association between tumor necrosis factor-α and chronic obstructive pulmonary disease: A systematic review and meta-analysis. Therapeutic Advances in Respiratory Disease. 2019;13:1753-1759.

Suissa S, Ernst P, Hudson M. TNF-α antagonists and the prevention of hospitalisation for chronic obstructive pulmonary disease. Pulmonary Pharmacology & Therapeutics. 2008; 21(1):234-238.

George PM, Wells AU, Jenkins RG. Pulmonary fibrosis and COVID-19: the potential role for antifibrotic therapy. The Lancet Respiratory Medicine. 2020;8(8): 807-815.

Lechowicz K, Drożdżal S, Machaj F, Rosik J, Szostak B, Zegan-Barańska M, Kotfis K. COVID-19: the potential treatment of pulmonary fibrosis associated with SARS-CoV-2 infection. Journal of Clinical Medicine. 2020;9(6):1917 - 1923.

Hanna A, Frangogiannis NG. Inflammatory cytokines and chemokines as therapeutic targets in heart failure. Cardiovascular Drugs and Therapy. 2020;34(6):849-863.

Harder-Lauridsen NM, Krogh-Madsen R, Holst JJ, Plomgaard P, Leick L, Pedersen BK, Fischer CP. Effect of IL-6 on the insulin sensitivity in patients with type 2 diabetes. American Journal of Physiology-Endocrinology and Metabolism. 2014;306(7):769-778.

Reid MB, Li YP. Tumor necrosis factor-α and muscle wasting: A cellular perspective. Respiratory Research. 2001;2(5): 1-4.

Mavrogonatou E, Konstantinou A, Kletsas D. Long-term exposure to TNF-α leads human skin fibroblasts to a p38 MAPK-and ROS-mediated premature senescence. Biogerontology. 2018; 19(3):237-249.

Omarjee L, Janin A, Perrot F, Laviolle B, Meilhac O, Mahe G. Targeting T-cell senescence and cytokine storm with rapamycin to prevent severe progression in COVID-19. Clinical Immunology. 2020;216:108464.

Bischof E, Siow RC, Zhavoronkov A, Kaeberlein M. The potential of rapalogs to enhance resilience against SARS-CoV-2 infection and reduce the severity of COVID-19. The Lancet Healthy Longevity. 2021;2(2):105-111.

Hadid T, Kafri Z, Al-Katib A.). Coagulation and anticoagulation in COVID-19. Blood Reviews. 2021;47:100761.

Serebrovska ZO, Chong EY, Serebrovska TV, Tumanovska LV, Xi L. Hypoxia HIF-1α and COVID-19: from pathogenic factors to potential therapeutic targets. Acta Pharmacologica Sinica. 2020;41(12):1539-1546.

Gralinski LE, Bankhead III A, Jeng S, Menachery VD, Proll S, Belisle SE, Baric RS. Mechanisms of severe acute respiratory syndrome coronavirus-induced acute lung injury. The American Society for Microbiology. 2013;4(4):271 - 279.

Yu M, Nardella A, Pechet L. Screening tests of disseminated intravascular coagulation: Guidelines for rapid and specific laboratory diagnosis. Critical Care Medicine. 2000;28(6):1777-1780.

Corman V. M, Landt O, Kaiser M, Molenkamp R, Meijer A, Chu DKW. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveillance. 2020;25(3):1-8.

U.F.A.D. Administration. Acclerated Emergency Use Authorization (EUA) Summary Orig3n 2019 Novel Coronavirus (COVID-19) Test. ORIG3N INC; 2020. Available:https://www.fda.gov/media/136873/download

(accessed May 2020)

Dsw DU, WuWu IS. Social welfare services and broken homes: The role of social work practice in Eleme Local Government Area of Rivers State. Central Asian Journal of Social Sciences and History. 2021;2(8):117-131.

Kotrlik JWKJW, Higgins CCHCC. Organizational research: Determining appropriate sample size in survey research appropriate sample size in survey research. Information Technology Learning and Performance Journal. 2001;21(1):43.

Chiedozie AP, Chukwuebuka OJ, Chidimma CF, Onyinyechi OV, Chijioke AK, Chibuzor OS, Chioma UB. Willingness to accept a potential COVID-19 vaccine in Nigeria. American Journal of Medical Science. 2021;9(1):1-5.

Arya M, Shergill IS, Williamson M, Gommersall L, Arya N, Patel HR. Basic principles of real-time quantitative PCR. Expert Review of Molecular Diagnostics. 2005;5(2):209-219.

Cheng VC, Lau SK, Woo PC. Yuen KY. Severe acute respiratory syndrome coronavirus as an agent of emerging and reemerging infection. Clinical Microbiology Review. 2007;20(4):660-694.

McBride CS, Baier F, Omondi A. B, Spitzer SA, Lutomiah J, Sang R, Ignell R, Vosshall LB. Evolution of mosquito preference for humans linked to an odorant receptor. Nature. 2014;515 (7526):222-227.

Zafer MM, El-Mahallawy HA, Ashour HM. Severe COVID-19 and sepsis: Immune pathogenesis and laboratory markers. Microorganisms. 2021;9(1):159-164.

Chaolin H, Yeming W, Xingwang L. Clinical features of patients infected with 2019 novel coronavirus in Wuhan China. Lancet. 2020;1(395):497-506.

Adias TC, Eze EM, Green MB. Evaluation of chemokine profile in stored whole blood. Journal of Medical Science and clinical Research. 2018;6(06):978-983.