Casanova et al demonstrated fecally contaminated liquid droplets of SARS cases are a potential vehicle for contagion besides respiratory droplets. This study reported that SARS coronaviruses remained infectious in water and sewage for days to weeks.[1]
For this reason, waterborne infection of COVID-19 coronaviruses should be checked meticulously to prevent further spread of COVID-19.
In addition to facial masks for blocking respiratory droplets, drinking water should be sterilized. At home, we must boil water or mix bitter herbs or lemons with water before drinking.
The government should pay attention to sterilize water supply source.
REFERENCE:
Casanova L, Rutala WA, Weber DJ, Sobsey MD. Survival of surrogate coronaviruses in water. Water Res. 2009 Apr;43(7):1893-8.
Conventional coronavirus is known as an ss(single-stranded) RNA virus. Its incubation period is 2 ~ 5 days, whereas novel coronavirus is 4 ~ 14 days and SARS one is 2 ~ 7 days.
As supportive evidence of the incubation period of novel coronavirus longer than that of SARS coronavirus, there was a report three persons recovered from COVID-19 still had positive RT-PCR results for novel coronaviruses, even without symptoms and contagion into nearby family members.[1]
Its clinical implication has not been solved.
Despite novel coronaviruses are similar to SARS ones, why is the incubation period of novel coronaviruses longer than SARS ones?
What are the factors to explain the above phenomena?
In SARS, viral shedding is the nonlytic release of the vast majority of mature virions into the extracellular space via the Golgi apparatus from the ER.[2]. If novel coronavirus behaves in this way, the incubation period may be similar to that of SARS coronavirus.
If so, there must be the other pathway for viral shedding and replication to be continued except for the extracellular space, namely the intracellular space.
Which compartment of the intracellular space is probably suitable for viral shedding and replication?
The author supposes that the isolated hidden place for introverted viral shedding into the cell may be an intranuclear space, for the vast majority of viral release happens on the extracellular cytoplasmic border, as aforementioned.
The author infers the answer from the point that the main target cell organelle by novel coronavirus is endoplasmic-reticulum(ER).
Among four main proteins in coronaviruses, N protein forms a complex by binding to genomic RNA and M protein triggers the formation of interacting virions in this endoplasmic reticulum-Golgi apparatus intermediate compartment(ERGIC) with this complex.[3][4]
The characteristic electron microscopic findings of the alveolar epithelial cells in SARS cases are markedly dilated rough endoplasmic reticulum(RER) and smooth endoplasmic reticulum(SER).[5]
Uniquely, there was a comment on the membranous inclusion bodies in some nuclei.[5]
Considering the genetic & electron microscopic similarity of novel coronavirus to SARS coronavirus, the chance for nuclear inclusions to occur is high.
About the structure of nuclear inclusions, the reference journal did not include the related figure, and we can not clearly describe the electron microscopic feature to explain viral shedding into the intra-nuclear space.
Instead, the electron microscopic finding of nuclear inclusions in the cells may be an alternative tool to explain why nuclear inclusions in novel coronaviral infected cells can be a hidden reservoir for persistent viral replication.
About the detailed structure of nuclear inclusion, that in pituitary adenoma may be appropriate for a comparable explanation.
The nuclear inclusion in pituitary adenoma is composed of the ER-rich cytoplasm within the cell nucleus.
Yang et al(2003) analyzed the genesis of nuclear inclusion as the result of intracytoplasmic invagination into the nucleus. [6]
A part of the outline of the nuclear inclusion is composed of a double-layered membrane at the locus between the nucleus and the cytoplasm. Its origin can not be proved to be from whether ER or nuclear membrane.
The point is that nuclear inclusion includes ER-rich cytoplasmic components, which is continuous with the ER on the cytoplasmic side.
The author thinks that this nuclear inclusion with ER-rich cytoplasmic component can be a reservoir for persistent viral replication even during viral release into the extracellular space and cell division.
Despite the detrimental cytolytic cytotoxicity in COVID-19 cases, the presence of the long incubation period of novel coronaviruses can be explained by this hypothesis.
Hence, further research for novel coronaviruses should be performed to investigate the presence of nuclear inclusions in COVID-19 cases.
P.S.
If nuclear inclusions in COVID-19 cases will be found, please correspond to me about the next step for therapeutic strategy.
REFERENCE:
[1] Lan L, Xu D, Ye G, et al. Positive RT-PCR Test Results in Patients Recovered From COVID-19. JAMA. Published online February 27, 2020.
[2] Denison MR. CORONAVIRUS RESEARCH: KEYS TO DIAGNOSIS, TREATMENT, AND PREVENTION OF SARS. In: Institute of Medicine (US) Forum on Microbial Threats; Knobler S, Mahmoud A, Lemon S, et al., editors. Learning from SARS: Preparing for the Next Disease Outbreak: Workshop Summary. Washington (DC): National Academies Press (US); 2004.
[3] de Haan CA, Masters PS, Lili Kuo, Harry Vennema, Peter JM, Rottier. Coronavirus particle assembly: primary structure requirements of the membrane protein. J Virol. 1998; 72: 6838-6850.
[4] Escors D, Ortego J, Enjuanes L. The membrane M protein of the transmissible gastroenteritis coronavirus binds to the internal core through the carboxy-terminus. Adv Exp Med Biol. 2001; 494: 589-593.
[5] Ding Y, Wang H, Shen H, Li Z, Geng J, Han H, Cai J, Li X, Kang W, Weng D, Lu Y, Wu D, He L, Yao K. The clinical pathology of severe acute respiratory syndrome (SARS): a report from China. J Pathol. 2003 Jul;200(3):282-9.(PDF free download)
[6] Yang SW, Yang KM, Kang Hy, Kim TS. Intranuclear cytoplasmic pseudoinclusions in pituitary adenomas. Yonsei Med J. 2003 Oct 30;44(5):816-20.(PDF free download)
te Velthuis et al reported that Zn(2+) inhibits the replication of SARS coronaviruses in cell culture study. They found that the SARS-CoV RdRp elongation was inhibited and template binding reduced by Zn(2+).[1]
Mocchegiani et al found that old people aged 60-65 years with specific IL-6 polymorphism (GG allele carriers; named C-) had a low level of zinc and the proinflammatory status of IL-6. With zinc supplementation, the inflammatory responses by IL-6 were ameliorated into the healthy condition.[2]
Old people have a tendency to have an increased level of IL-6 associated with the aging process.[3]
It has known that increased IL-6 production by Th2 cells and macrophages.[4]
This Th2 shifting immunity is susceptible to viral infection.
About the immunity of COVID-19, Xu et al revealed Th2 immune skewness polarized by IL-6 and TGF-β, which is vulnerable to viral infection.[5]
With a constellation of the above data, although there is no report about the relationship between zinc and novel coronaviral infection(COVID-19, Wuhan Coronaviral infection), the author thinks that zinc can be a supportive nutritional mineral for anti-viral immunity in COVID-19.
REFERENCE:
[1] te Velthuis AJ, van den Worm SH, Sims AC, Baric RS, Snijder EJ, van Hemert MJ. Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture. PLoS Pathog. 2010 Nov 4;6(11):e1001176.
[2] Mocchegiani E, Romeo J, Malavolta M, Costarelli L, Giacconi R, Diaz LE, Marcos A. Zinc: dietary intake and impact of supplementation on immune function in elderly. Age (Dordr). 2013 Jun;35(3):839-60. doi: 10.1007/s11357-011-9377-3. Epub 2012 Jan 6. PMID: 22222917; PMCID: PMC3636409.
[3] Franceschi C. Inflammaging as a major characteristic of old people: can it be prevented or cured? Nutr Rev. 2007 Dec;65(12 Pt 2):S173-6.
[4] Mocchegiani E, Muzzioli M, Cipriano C, Giacconi R. Zinc, T-cell pathways, aging: role of metallothioneins. Mech Ageing Dev. 1998;106:183–204.
[5] Xu Z, Shi L, Wang Y, Zhang J, et al. Pathological findings of COVID-19 associated with acute respiratory distress syndrome. Lancet Respir Med. 2020;S2213-2600(20)30076-X.
Chapter 12 of Exodus in Bible reports on Passover day. Here God ordered the Israelite to avoid disaster on Passover day.
Chapter 12: verse 12th and 13th of Exodus, Bible says, "12on that same night I will pass through Egypt and strike down every firstborn - both men and animals - and I will bring judgment on all the gods of Egypt. I am the LORD.13The blood will be a sign for you on the houses where you are; and when I see the blood, I will pass over you. No destructive plague will touch you when I strike Egypt" (Figure 1).
The Passover means the disaster skipped the Israelite who kept the orders of God.
God orders the Israelite to paint the blood of lamb or goat on the sides and tops of the doorframe.
Why did God order this?
I interpreted this question medically. In terms of pathology, my opinion on these two problems is as follows;
Blood color varies from red to dark purple according to oxygen saturation. The main component of blood hue is caused by RBC (Red Blood Cell). When oxygen is rich in blood, blood color is red by binding O2to iron in hemoglobin. Conversely, when oxygen is deficient, blood hue is dark purple. When we have cut wound in the skin, the blood coagulates slightly sticky. This occurs mainly by the fibrin component. The fibrin particles are interlocked and form chains of fibrin, a structure of meshwork (Fig.2.). This meshwork structure filters blood cells, especially platelets and RBCs. All this process leads to blood coagulation. The characteristics of the fibrin component endow the blood glutinous.
Sticking nature of blood by fibrin and blood other components, namely viscous force, makes blood to acquire the filtering power to capture fine dust of air including microorganisms (bacteria and virus particles).
This may dilute the density of microbial pathogen particles.
As a principle of vaccine, the diluted amount of microbial pathogens may work as an immune-boosting effect rather than causing disease.
In this situation, I infer the people who did not paint blood on the doorframe may inhale contaminated air and be exposed to the infectious state. In contrast, people, namely the Israelites who paint blood on the doorframe, may inhale the air of diluted infectious microbial particles, which worked as a vaccine effect.
Furthermore, God designates that doorframe painting with blood should be done at night (Exodus, chapter 12; verse 6th:Take care of them until the fourteenth day of the month, when all the people of the community of Israel must slaughter them at twilight). To keep the viscosity of blood, the blood should preserve the liquidness. The liquid state may most be maintained at the cool state. The cool state in a day may be kept at the night time after sunset.
For the above reason, I infer God ordered blood paint on the doorframe at night in a day.
Copyright (C-2013-017025)PASSOVER DAY – Medical Interpretation by a Pathologist – (English)
Facial Mask in the year 2020:
As of March. 2020, novel coronavirus infection is pandemic. As a primary preventive method, the importance of the facial mask is raised. The author thinks that the facial mask may play a role in decreasing the number of viral droplets in inflowing air like the doorframe painted with blood in Passover.
Human coronaviruses have a property to haemagglutinate with sheep erythrocytes(red blood cells).[1] In contrast, human coronaviruses do not haemagglutinate with chick erythrocytes.[2]
Haemagglutination of sheep RBCs by viruses
These findings are reminiscent of the interpretaton by Yang(2013) about 'Passover and Blood of Sheep or Goat'.[3]
The story about 'Passover and Blood of Sheep or Goat' is below:
In chapter 12: verse 5th ~ 7th of Exodus, Bible says, "5The animals you choose must be year-old males without defect, and you may take them from the sheep or the goats. 6; Take care of them until the fourteenth day of the month, when all the people of the community of Israel must slaughter them at twilight.7Then they are to take some of the blood and put it on the sides and tops of the doorframes of the houses where they eat the lambs".
There is a question, "why did God designate the blood of lamb or goat for doorframe painting?”.
In that time of Exodus, there should be poultry and animals besides sheep and goat.
If the stickiness of blood is enough for removing infectious microbial pathogens, why did God designate specifically the blood of lamb or goat?
Concerning this question, I think the blood painted on doorframe even at night can be dry in some degree. In the partial dry state, the microbial organisms attached within blood can be detached and easily floated into the air. The microbial-contaminated air can be inhaled into the lung of the Israelite and cause an infectious state.
To overcome this problem, the blood of lamb or goat should have a certain specific attribute to seize microbial organism, even in the dry state of the blood.
Regarding this problem, there is an interesting report by Medeiros et al (2001. Virology 289:74-85).[4] Medeiros et al found influenza virus particles stick to the RBCs (Red Blood Cell) in the blood of lamb or goat and aggregates RBCs (termed 'agglutination'). In contrast, the RBCs in the blood of chicken does not cause this reaction. As inferred from this fact, there must be specificity in each species of animal or poultry, in RBC agglutination to particular microbial pathogens.
So I think God may order the blood of lamb or goat to remove the particular microbial pathogens at the Passover day night.
Through the stickiness and the RBC agglutination of the blood of lamb or goat, it is more likely that the amount of filtered microbial pathogens may work as a vaccine effect.
In the chapter 12 of Exodus in the old testament, verse 12th and 13th, the Bible says, "12 on that same night I will pass through Egypt and strike down every firstborn - both men and animals - and I will bring judgment on all the gods of Egypt. I am the LORD.13The blood will be a sign for you on the houses where you are; and when I see the blood, I will pass over you.
Although there can be other pathogens and view points to be considered, the plausible facts of haemagglutination with sheep erythrocytes and the rapid occurrence of death overnight seem to be similar to those of COVID-19 infection.
REFERENCE:
1. Hierholzer JC, Tannock GA. Quantitation of antibody to non-hemagglutinating
viruses by single radial hemolysis: serological test for human coronaviruses. J Clin Microbiol. 1977 Jun;5(6):613-20.
2. Gerdes JC, Klein I, DeVald BL, Burks JS. Coronavirus isolates SK and SD from multiple sclerosis patients are serologically related to murine coronaviruses A59 and JHM and human coronavirus OC43, but not to human coronavirus 229E. J Virol. 1981 Apr;38(1):231-8.
3. Yang SW. Episode 2.Passover and Blood of Sheep or Goat.PASSOVER DAY – Medical Interpretation by a Pathologist (English).Copyright (C-2013-017025)
4.Medeiros R, Escriou N, Naffakh N, Manuguerra JC, van der Werf S.Hemagglutinin residues of recent human A(H3N2) influenza viruses that contributeto the inability to agglutinate chicken erythrocytes. Virology. 2001 Oct 10;289(1):74-85.
Pleomorphic, ranging from 60 to 220 nm in diameter
ETYMOLOGY
(Latin) corona = crown.
APPEARANCE
A fringed appearance of widely spaced club-spaced surface projections, viral spike(S) peplomers, reminiscent of a solar corona.
PROTEINS that contribute to the overall structure of coronaviruses
Spike (S), envelope (E), transmembrane (M), and nucleocapsid (N)
OUTLINE of coronaviruses
Envelope composed of lipid bilayer.
GENOME
Non-segmented single-stranded positive-sense RNA of approximately 30 kb
PROPERTY
The ability to haemagglutinate
Spike (S) protein
1. Virions(infected form of the virus) can be bound to specific surface receptors in the plasma membrane of the host cell via the N-terminus of the S proteins [9]
2. The major inducer of neutralizing antibody
Envelope (E) protein
A small, integral membrane protein involved in several aspects of the virus’ life cycle, such as assembly, budding, envelope formation, and pathogenesis.[2]
Transmembrane (M) protein
1. The virion to fuse into the cell and to make protein antigenic.[3][4][5]
2. N protein forms a complex by binding to genomic RNA and M protein triggers the formation of interacting virions in this endoplasmic reticulum-Golgi apparatus
intermediate compartment (ERGIC) with this complex.[6][7][8]
Nucleocapsid (N) protein
Virion structure, replication and transcription of coronaviruses.
Fig. Coronavirus Structure
REFERENCE
[1] David Greenwood, Richard C. B. Slack, John F. Peutherer. MEDICAL MICROBIOLOGY. 16 ed. 2002. Churchill Livingstone.
[2] Schoeman, D., Fielding, B.C. Coronavirus envelope protein: current knowledge.Virol J16,69 (2019).
[3] de Haan CA, de Wit M, Kuo L, Montalto-Morrison C, Haagmans BL, Weiss SR, et al. The glycosylation status of the murine hepatitis coronavirus M protein affects the interferogenic capacity of the virus in vitro and its ability to replicate inthe liver but not thebrain. Virology. 2003; 312:395-406.
[4] Alexander S, Elder JH. Carbohydrate dramatically infl uences immune reactivity of antisera to viral glycoprotein antigens. Science. 1984; 226: 1328-1330.
[5] Wissink EH, Kroese MV, Maneschijn Bonsing JG, Meulenberg JJ, van Rijn PA, Rijsewijk FA, et al. Signifi cance of the oligosaccharides of the porcine reproductive and respiratory syndrome virus glycoproteins GP2a and GP5 for infectious virus production. J Gen Virol. 2004; 85: 3715-3723.
[6] de Haan CA, Masters PS, Lili Kuo, Harry Vennema, Peter JM, Rottier. Coronavirus particle assembly: primary structure requirements of the membrane protein. J Virol. 1998; 72: 6838-6850.
[7] Escors D, Ortego J, Enjuanes L. The membrane M protein of the transmissible gastroenteritis coronavirus binds to the internal core through the carboxy-terminus. Adv Exp Med Biol. 2001; 494: 589-593.
[8] Narayanan K, Makino S. Characterization of nucleocapsid-M protein interaction in murine coronavirus. Adv Exp Med Biol. 2001; 494:577-582.
[9] Lewicki DN, Gallagher TM. Quaternary structure of coronavirus spikes in complex with carcinoembryonic antigen- related cell adhesion molecule cellular receptors. J Biol Chem. 2002; 277: 19727-19734.
70% ethanol reduced viral infectivity below the detectable level.
There can be a question about how ethanol inactivates viral particles.
About this, Toppozine et al(2012) reported the evidence for enhanced permeability in both fluid and gel phases of thephospholipidbilayers in the presence ofethanolmolecules.
The outline of Coronaviruses is composed of envelope structure, which is composed of lipid bilayer. The permeability of this lipid can be increased by ethanol.
For this reason, ethanol as a disinfectant for Coronaviruses can be useful.
REFERENCE:
1. Kariwa H, Fujii N, Takashima I. Inactivation of SARS coronavirus by means ofpovidone-iodine, physical conditions and chemical reagents. Dermatology.2006;212 Suppl 1:119-23. doi: 10.1159/000089211. PMID: 16490989.Date: 2020.03.05
2. Laura Toppozini, Clare Armstrong, Matthew A. Barrett, Songbo Zheng, Lindy Luo, Hirsh Nanda, Victoria Garcia Sakai, Maikel C. Rheinstadter. Partitioning of ethanol into lipid membrane and its effect on fluidity and permeability as seen by X-ray and neurtron scattering. Soft Matter, issue 47, 2012.
