Influenza - Science topic

Hello

1. Why this system does not recognise copies of same publication and remove them?

2. Also: My article on influenza and Pneumococcal vaccination was published in December and is my latest publication, yet your system decided from its head to make it published in January 24 and put it down in the list. Why is that?

Would the removal of methionine and alanine from the N-terminus of the Influenza A H3N8 nucleoprotein affect its function or structure, considering the goal of leaving a serine residue to create an optimal TEV protease cleavage site? My objective is to purify the nucleoprotein for use as an antigen in developing ELISA kits for diagnostic purposes, and I want to ensure that this modification does not compromise the protein's integrity or its suitability for diagnostic applications.

I have done 3 blind passages to concentrate the viral stock in the supernatant without adding TPCK Trypsin. After passage 1 my Ct value for INF A was 36.96. After P2 the value decreased to 33.36. However, I was expecting a much lower value in the mid-20s. I am freeze-thawing the flasks at -20 degrees to lyse the cells and release the virus into the medium. Is it the correct approach? and can I get a much higher titre without adding TPCK Trypsin?

Hi :)

How should I design primers for (-)ssRNA samples from Influenza A and hRSV viruses?

I need to select the protein, locate the gene sequence, and then use it to design primers. Should I retrieve the cDNA sequence from NCBI to work with?

The term 'cds' in FASTA means that it is the sequence without introns. Is it okay to use this type?

if you have any guides, videos or documents you would like to share with me, I would be very grateful.

Thanks

Dear Researchers:

Could you please share some simple cures or prevention for COVID-19, Cold, Flu or Influenza, and possibly Other Viruses, and Cancers?

Updates on Oct. 10, 2023: First, many thanks to all contributors to this discussion. Here are some Natural Approaches found from surveying literature in medicine to Boost our Immune Systems against viruses such as COVID-19, Cold, Flu or Influenza infections and to avoid/minimize developing further inflammations in the lungs and hearts caused by some of those viruses:

Give it a try, please! Especially if you increase your Vitamin D level to a required level and consume Vitamin C sources, e.g., oranges, on a daily basis, you can check how rarely you would catch the virus. Or, even after catching the virus, the virus will likely develop very mild symptoms in your body.

1- Daily uptake of Vitamin D pills up to 100 IU per 1 kg weight is safe and very important, recommended by Afshar et al. (2000) and Dr. Hamid Sajjadi in an interview, to RAISE the Vitamin D level in our body to the POINT which is REQUIRED to BOOST our IMMUNE SYSTEMS against Viruses and Diseases including Cancers.

Please read the following article by Afshar et al. (2000) about the importance of vitamin D and the required daily dose of it (Up to 100 IU per 1 kg weight) to boost our Immune Systems.

Please also read the following Review article by Jordan et al. (2022) about the importance of Vitamin D on the level of infection & disease progression for COVID-19. You may find in the article the importance of our Forgotten SUN.

Vitamin D is rarely available in food sources, except in fatty fish which needs to be eaten high enough to get the required amount of Vitamin D for a body.

Another good natural source is daily sunbathing with naked skin; however, in cloudy regions such as Europe, sunbathing doesn't work well.

Vitamin D helps to absorb Calcium in our intestines and thus, in order to avoid excessive absorption of Calcium by our body, it would be better to use Vitamin D pills with Calcium sources such as warmed-up milk and Magnesium sources such as bananas on a daily basis. Because magnesium competes with calcium in our intestines to get absorbed.

Here is a text from A Review article by Kulie et al. (2009) about some of the importance of Vitamin D on our health:

"Vitamin D is a fat-soluble vitamin that plays an important role in Bone Metabolism and seems to have some Anti-Inflammatory and Immune-Modulating properties. In addition, recent epidemiologic studies have observed relationships between low vitamin D levels and multiple disease states.

Low vitamin D levels are associated with increased overall and Cardiovascular mortality, Cancer incidence and mortality, and Autoimmune Diseases such as Multiple Sclerosis. Although it is well known that the combination of vitamin D and calcium is necessary to maintain Bone Density as people age, vitamin D may also be an independent risk factor for falls among the Elderly."

2- Having Good Nutrients including Protein sources, Minerals, and Other Vitamins, e.g., C, A, and E, sources from fresh fruits, vegetables, and nuts. For example, the good sources of fruits and vegetables for these vitamins could be a daily use of 1-2 Oranges for Vitamin C, Carrots for Vitamin A, and Almonds or Sunflower Seeds for Vitamin E.

As Vitamin C is a water-soluble vitamin, the excess of it will be excreted from the body, it needs to be consumed every day to provide everyday vitamin C requirements for the body, as it is the 2nd most important vitamin after Vitamin D to boost our Immune Systems against viruses and diseases.

And, Vitamin B family from grains, poultry, and meat sources.

3- After the infection by those viruses, gargling salty water to disinfect the throat to avoid further movement of the virus into the lungs as the virus may stay in the throat for a few days

4- Inhaling Steamed Fresh Leaves, if not available, the Oil, of Eucalyptus 4-5 times a day for several continuous days to kill the virus in the lungs.

Here is A Review article by Mieres-Castro et al. (2021) about the "Antiviral Activities of Eucalyptus Essential Oils: Their Effectiveness as Therapeutic Targets against Human Viruses"

Australian Aboriginals are very much using Eucalyptus to Treat Infections.

5- Having plenty of Warm Drinks to wash out the virus from our body and dilute the blood to avoid blood clotting.

6- Having enough sleep and daily activities/exercises

7- Kids are proven to have High Immunity Against COVID-19, likely due to having a high amount of Melatonin, the Sleep Hormone, in their blood. So, that is why kids sleep very much as you know.

Melatonin production in our body usually decreases with increasing age. Thus, we may use daily melatonin pills after the infection based on what physicians may prescribe for us.

Here is A Review article by Carrillo-Vico et al. (2013) about the Importance of Melatonin on the Functionality of Our Immune Systems:

8- Avoid Fear/Panic as it Substantially Deteriorates the Functionality of Immune Systems against viruses and diseases.

Here is an interview by Dr. Lauren Deville about How Fear Affects Our Immune System:

This RG open question is linked to the previous about the dramatic evolution (partly unexplainable) of COVID19 in Northern Italy during wave 1.

The previous RG open question is reported below🔴 and resulted in a completely alternative model for the evolution🟨 of SARS-CoV/2 from pre-pandemic phase to pandemic phase.

In this specific RG question, the intention is to create an open discussion on the possible emergence of a violent outbreak of avian flu or similar in central Europe.

This concern arises from a qualitative model that links three events which in the past have always characterized the violent explosion of a bird flu or similar.

The model involves three steps.. https://www.researchgate.net/publication/367046404

This RG open question will serve to accumulate data both for and against this dire possibility.

Thanks to all the participants.

|--sv--|

🔴The novel Coronavirus in N. Italy, Lombardia 【 COVID19 / 2019nCoV / SARSCoV2 】 shows a fatality rate compatible with SARS-MERS. Why?? MAR.2020. -- https://www.researchgate.net/post/The-novel-Coronavirus-in-N-Italy-Lombardia-COVID19-2019nCoV-SARSCoV2-shows-a-fatality-rate-compatible-with-SARS-MERS-Why

🟨Link between the start of pandemic SARS-CoV/2 (COVID19) and the Huanan Seafood Wholesale Market in Wuhan (Hubei: China): the furin cleavage site of spike protein. FEB.2022. -- https://www.researchgate.net/publication/358443761

It is common during the season to increase the incidence of severe influenza, the symptoms of which are similar to Corona 19. In your opinion, is the weather the main cause, or is it the results of infection with Corona, or is it possible that a new variant is currently developing? Are there recent studies in Iraq on this subject?

Covid news – live: Cases soar again in India as doctors warn of ‘new symptom’

A new coronavirus strain dubbed Arcturus appears to be driving a surge in Covid-19 cases in India, prompting the country to resume vaccine production and sparking fears it could lead to a rise in cases in the UK and elsewhere.

India on Friday recorded 11,109 new Covid infections, the biggest jump in almost a year. The country’s active case count is now up to 49,662.

The XBB.1.16 strain, a sub-variant of Omicron, has been found in 22 countries, including Singapore, Australia, the UK and the US. Research indicates Arcturus could be one 1.2 times more infectious than the last major sub-variant, making it likely to become the dominant strain.

The spread of the strain, first detected in late January in India, is worrying experts, as it seems to exhibit unique symptoms in children, one of which is conjunctivitis.

The symptoms of the variant include high fever, cough, and “itchy” conjunctivitis or pinkeye, according to Vipin Vashishtha, a paediatrician and former head of the Indian Academy of Pediatrics Committee on Immunisation.

source: https://ca.yahoo.com/news/covid-news-live-cases-soar-075336169.html

COVID-19 and Influenza Activity

April 2, 2023 to April 8, 2023

These images provide a high-level assessment of respiratory virus activity in Ontario. Provincial percent positivity can be used to provide an estimate of the intensity of circulating viruses in the province. Percent positivity for the most recent week is used to assign influenza and COVID-19 to either a low, moderate, high or very high category. Weekly indicator change was determined by considering a combination of indicators (see Technical Notes). For further details, please refer to the Respiratory Virus Overview in Ontario report.

source: https://www.publichealthontario.ca/en/data-and-analysis/infectious-disease/covid-19-data-surveillance/covid-19-data-tool?tab=overview

According to most U. S. state health department records the levels of influenza recorded during the 2020-2021 flu season and the 2021-2022 flu season, when Covid 19 surges occurred, the levels of infection by influenza was below the baseline. Even the national data from the CDC shows the same thing, but no explanation has come forward to explain this phenomenon.

I have a question. Do prior immunity to a virus-like particle vaccine platform affects its immunogenicity for the next vaccination? For example, if we vaccinate an individual using a chimeric VLP consist of an antigen of interest fused into influenza M1 protein as the core, will it have a lower immune response if the individual were vaccinated with other VLP-based vaccine containing the same M1 protein? Will it happen as with the viral vector vaccine?

Hi Dear researcher

Some Influenza subtypes are recommended for vaccination by WHO. Unfortunately the current database of influenza have not update sequences about influenza. could you help me to find the protein sequences of following subtypes?

H1N1 A/Sydney/5/2021 (H1N1)pdm09-like virus

H3N2 A/Darwin/6/2021 (MDCK-SIAT derived)

B/Austria/1359417/2021 (MDCK derived)

I am wondering if the use of collagen as a matrix on the plate, or other matrix, with the consequent cell polarization, is necessary to get good virus yields. Many studies does not report culture on collagen or other matrix.

I plan to use BALB/c mice for studying pneumonia caused by the mouse-adapted influenza virus (A/PR8/H1N1).

I want to know, what is the requirement to determine the sex of an animal? ​It seems that most of them choose female mice, but there are also researchers who use half male and female mice. What is the difference between them? Does the sex of the mouse have a significant impact on the final result?

ZhiYong Peng, Zhongnan Hospital

Brief Summary:

2019 new coronavirus (2019-nCoV) infected pneumonia, namely severe acute respiratory infection (SARI) has caused global concern and emergency. There is a lack of effective targeted antiviral drugs, and symptomatic supportive treatment is still the current main treatment for SARI.

Vitamin C is significant to human body and plays a role in reducing inflammatory response and preventing common cold. In addition, a few studies have shown that vitamin C deficiency is related to the increased risk and severity of influenza infections.

We hypothesize that Vitamin C infusion can help improve the prognosis of patients with SARI. Therefore, it is necessary to study the clinical efficacy and safety of vitamin C for the clinical management of SARI through randomized controlled trials during the current epidemic of SARI.

Vitamin C, also known as ascorbic acid, has antioxidant properties. When sepsis happens, the cytokine surge caused by sepsis is activated, and neutrophils in the lungs accumulate in the lungs, destroying alveolar capillaries. Early clinical studies have shown that vitamin C can effectively prevent this process. In addition, vitamin C can help to eliminate alveolar fluid by preventing the activation and accumulation of neutrophils, and reducing alveolar epithelial water channel damage.

At the same time, vitamin C can prevent the formation of neutrophil extracellular traps, which is a biological event of vascular injury caused by neutrophil activation. Vitamins can effectively shorten the duration of the common cold. In extreme conditions (athletes, skiers, art workers, military exercises), it can effectively prevent the common cold.

And whether vitamin C also has a certain protective effect on influenza patients, only few studies have shown that vitamin C deficiency is related to the increased risk and severity of influenza infections. In a controlled but non-randomized trial, 85% of the 252 students treated experienced a reduction in symptoms in the high-dose vitamin C group (1g / h at the beginning of symptoms for 6h, followed by 3 * 1g / day).

C19 tests may show cross reactivity or interference with other viruses

CR: human coronavirus 229E, human coronavirus OC43, human coronavirus NL63, MERS coronavirus

IF: Influenza A, B; RSV, Protein A-positive Staphylococcus aureus

Is there any laboratory which is capable to test these?

Kind regards,

Dietrich Doll

Do we need adapt influenza viruses in MDCK cells before conducting serum neutralization test with influenza viruses in MDCK cels?? Thanks

From 40 to 64 years of age, about 92,000 people have died or at least their deaths are attributed to Covid in the United States alone. https://www.statista.com/statistics/1191568/reported-deaths-from-covid-by-age-us/ If the new variants are as reported this disease is evolving to a more lethal version that kills younger patients https://www.cnbc.com/2021/03/11/covid-variant-in-the-uk-appears-to-be-64percent-more-deadly-than-other-strains-study-finds.html This begs serious questions did our defensive measures hand washing, masks, and quarantines encourage this trend to appear. Certainly, the modern practice of clustering or warehousing elderly members of society allowed the virus to specialize in rapid reproduction very similar to what happened to Spanish influenza in the trenches of WW1. Reports of unmitigated stress, binge eating, monthly weight gain lend credence that confinement might have been counterproductive. Indeed, some might argue that had we done no quarantining we would have gained herd immunity by now. Instead, are we giving Covid the time to evolve each time it breaks into a protected sector of the population? Please feel free to debate this question in the replies.

Dear Fellows

With the developing stories of the spread of COVID-19 all around the world, which has been declared as a pandemic by WHO recently, I am wondering, in how much time this virus would vanish from the surface of the Earth? Is there any scientific study available for this?

Please share your opinions.

Hello,

I have recently started to do some Plaque assays to determine Influenza titer. However, my plaques dont really look nice and are therefore hard to count, so I wanted to ask if someone has some suggestions to improve that.

Some Info: I use MDCK-II cells in 96-well plates for infection and after a 1 hour infection period I incubate these for 28 hours with an 50/50 mixture of Avicel/2xMEM + Trypsin.

I have attached a picture of one of my assays so you can see that sometimes the layer of cells seems to be disrupted, making me unable to count properly.

Thanks for your help.

Hi all,

I wonder if anyone know where I can find the plasmid map for pHH21 that is commonly used in Influenza research?

Hypothesis: Interleukin 10 (IL-10) is a potent anti-inflammatory cytokine which can guide regarding optimal use of steroids and immunosuppressants, allowing immune system to fight at its best.

Recovery trial has shown positive effects of steroids in COVID-19 patients who were receiving either invasive mechanical ventilation or oxygen alone but not among those receiving no respiratory support¹.

Now-a-days steroids are among most widely used drugs in COVID pneumonia and we know that ‘steroid’ is a double-edged sword. If these drugs are given when patient is actually going through cytokine storm phase then it has beneficial effect but if we are administering them in appropriate immune response then it may be hazardous as it can cause suppression of immunity which in turn can escalate viral replication and subsequent disease progression.

There is no widely accepted definition of cytokine storm.

Cytokine storm is an umbrella term encompassing several disorders of immune dysregulation characterized by constitutional symptoms, systemic inflammation, and multiorgan dysfunction that can lead to multiorgan failure if inadequately treated².

IL-1, IL-6, ferritin, CRP are the part and partial of immune response irrespective of appropriate or dys-regulated.

Elevation of interleukin 6 and CRP is a part of cytokine storm but it’s not the absolute marker. We need to corelate these markers with overall clinical scenario to label it as cytokine storm.

So, which is the crucial moment to start steroid?

Interleukin 10 (IL-10) is a potent anti-inflammatory cytokine that plays a central role in limiting host immune response to pathogens, thereby preventing damage to the host and maintaining normal tissue homeostasis. It is also essential for regulation of immune responses³.

The generation of an effective immune response along with limiting tissue damage requires a delicate balance between pro- and anti-inflammatory responses³.

Impaired IL-10 expression and/or signaling can enhance clearance of pathogens during an acute infection, but also exaggerate inflammatory response, resulting in exacerbated tissue damage⁴.

Some pathogens can harness the immunosuppressive capacity of IL-10 to limit host immune response, resulting in persistent infection⁵.

Though absence of IL-10 is often initially beneficial, it’s prolonged deficiency can be detrimental in the long term. Prolonged and increased production of inflammatory cytokines can lead to septic shock in the context of viral, bacterial, or fungal infections^6,7.

Inflammatory molecules can often be potent activators of cell death and increasing levels of IL-10 can moderate the extent of apoptosis that is induced in response to infection.

For instance,

1. In a Chlamydia pneumoniae model, where bacterial clearance is enhanced in the absence of IL-10, mice also develop severe inflammation and experience elevated levels of apoptosis3.

2. During Mycobacterium avium infection, early IL-10 production is correlated with the failure of control of infection; ablation of IL-10 signaling led to enhanced pathogen control, demonstrating a causal relationship between IL-10 and the lack of pathogen control⁸.

3. In acute influenza infection, blocking the action of IL-10 results in enhanced pulmonary inflammation and harmful injury⁹.

It is not clear whether elevated IL-10 levels during infections are a cause or a consequence of high pathogen burdens, but studies indicate that resolution of infection requires a coordinated response in which initial pro-inflammatory mechanisms clear the pathogen and are subsequently limited by IL-10 before pathology occurs³.

A unique feature of the COVID-19 cytokine storm is the dramatic elevation of interleukin 10 (IL-10) in severe/critically ill patients which has led to speculation that IL-10 might play a pathological role in COVID-19 disease progression¹⁰.

1. If IL-10 levels are elevated along with IL-1, IL-6, CRP, ferritin etc.: This indicates that IL-10 is already trying to control the inflammation by acting as anti-inflammatory agent and in-turn might be helping virus to proliferate. If in such situation we are administering steroids to the patient which can further suppress immune response leading to further viral proliferation.

2. If IL-10 levels are suppressed along with elevated levels of IL-1, IL-6, CRP, ferritin etc: If patient is showing clinical symptoms and signs of cytokine storm then this is probably the best indicator of exaggerated inflammatory response and such patients are probably the best candidates to be benefited from steroids and immunosuppressants.

3. If IL-10 levels are normal along with elevated levels of IL-1, IL-6, CRP, ferritin etc: Use of steroids should be guided by clinical symptoms of signs of cytokine storm.

1. Lin WS, Emberson JR, Mafham M, Bell JL, Linsel L, Staplin N et al. Dexamethasone in Hospitalized Patients with Covid-19. N Engl J Med 2021;384:693-704. DOI: 10.1056/NEJMoa2021436

2. Fajgenbaum DC, June CH. Cytokine storm. N Engl J Med 2020;383:2255-73. DOI: 10.1056/NEJMra2026131.

3. Iyer SS, Cheng G. Role of Interleukin 10 Transcriptional Regulation in Inflammation and Autoimmune Disease. Crit Rev Immunol. 2012; 32(1): 23–63.

4. Li C, Corraliza I, Langhorne J. A defect in interleukin-10 leads to enhanced malarial disease in Plasmodium chabaudi chabaudi infection in mice. Infect Immun. 1999 Sep; 67(9):4435–42.

5. Brooks DG. Interleukin-10 determines viral clearance or persistence in vivo. Nature Med. 2006;12:1301–9.

6. Gazzinelli RT, Wysocka M, Hieny S, Scharton-Kersten T, Cheever A, Kühn R, Muller W, Trinchieri G, Sher A. In the absence of endogenous IL-10, mice acutely infected with Toxoplasma gondii succumb to a lethal immune response dependent on CD4+ T cells and accompanied by overproduction of IL-12, IFN-gamma and TNF-alpha. J Immunol. 1996 Jul 15; 157(2):798–805.

7. Li C, Corraliza I, Langhorne J. A Defect in inter-leukin-10 leads to enhanced malarial disease in Plasmodium chabaudi chabaudi infection in mice. Infection and Immunity. 1999; 67(9):4435–42.

8. Fiorentino DF, Bond MW, Mosmann TR. Two types of mouse T helper cell. IV. 32 clones secrete a factor that inhibits cytokine production by 31 clones. J Exp Med. 1989; 170:2081–95.

9. Sun J, Cardani A, Sharma AK, Laubach VE, Jack RS, Müller W, Braciale TJ. Autocrine regulation of pulmonary inflammation by effector T-cell derived IL-10 during infection with respiratory syncytial virus. PLoS Pathog. 2011 Aug.7(8):e1002173.

10. Lu L, Zhang H, Dauphars DJ, He YW. A Potential Role of Interleukin 10 in COVID-19 Pathogenesis. Trends in Immunology, January 2021, Vol. 42, No. 1.

Croup is a viral infection affecting the upper respiratory tract. Its causative organisms includes Influenza and Parainfluenza viruses. This question aims to know the role of antiviral agents in the management of disease.

The Spanish flu, also known as the 1918 influenza pandemic, was an unusually deadly influenza pandemic caused by the H1N1 influenza A virus. Lasting from February 1918 to April 1920, it infected 500 million people – about a third of the world's population at the time – in four successive waves. Deaths: 17–100 million (estimates)

One hundred years later - 2019 COVID-19 pandemic.137 million cases, 2 million 960 thousands deaths.

The Spanish flu started on hundred years ago and continued for two years. Had four waves.

The COVID-19 started 2019 and continued for now. Had two waves, but pandemic continues.

Can we expect third and forth wave of COVID-19 in shorth time future?

Hi, I am looking for a test lab that can do tests on anti-viral activity of formulations. Strains to be tested are HSV-1, and for other formulation typically HRV, Influenza, and covid-19.

test labs may be from the west, say US, EU, but also from India, China, indonesia.

We used to do the H1N1 WSN plaque assay without TPCK-trypsin, and it works fine. Now we are trying the H1N1 PR8 and H3N1 plaque assay, but the PR8 failed to form plaques, and the plaques of H3N1 was tiny and not clear.

I'm following the Virapur protocol, using DMEM and MDCK cells and my virus does make strong CPE effect in MDCK so I was wondering if adding TPCK-trypsin can optimised the result?

thank for the response 

Dear all,

I have a naive question. When we want to study Type 1 response, we infect mice with influenza for instance, if we want to study Type 2 response we use for instance T. Bruecei. Does anybody know which type of response we induce when we immunize with our classical protocols such as SRBC or NPCGG/NPOVA/NPKLH?

Thank you very much

As widely accepted presently, early childhood education and development are closely intertwined and Impossible to truly set apart. Adults, living in pandemic Times, are pivotal actors in the way this challenging Times integrate children's development.

Educators, parents, political leaders and other stakeholders how should we go about this to assure the best developmental outcomes for the new generation ranging from 2 to 6 year old, within this important frame of socialization and development stages? Should we look to what happened during the great influenza for lessons?

Previous research has suggested an involvement of meteorological conditions in the spread of droplet-mediated viral diseases, such as influenza. However, as for the recent novel coronavirus, few studies have discussed systematically about the role of daily weather in the epidemic transmission of the virus.

With the appearance of COVID-19 vaccines on the market, many available choices are there.

Which one is good?

Which one is safest?

Which one is most expensive?

Which one is easiest to store?

How many doses are required?

Other than intramuscular injection, any other forms?

How to check immune response after?

Do we need post-injection blood test?

Do we need annual booster dose?

Do we need new vaccines every year by prediction as if flu vaccines?

Any contraindications?

Any allergy from vaccination?

Do we need to mask after injection?

Hello,

Most hand sanitizers contain 60-80% alcohol. Alcohol destroys organisms including bacteria, fungi, protists, and enveloped viruses by breaking down the plasma membrane. In recent years, microbial communities on the skin, in the airways, and in the gut have been shown to play an important role in health. Does daily use of hand sanitizer negatively impact the diversity and strength of the hand microbiome? If so, what are the implications of this to overall health? Do skin microbes play beneficial roles such as helping to prevent transmission of other pathogens?

Thank you.

All literature data I came across showed that Ribavirin had equal if not greater effect in titer reduction, survivability, and body weight loss compared to Oseltamivir. The biggest difference I found was that Ribavirin caused lower levels of interleukins but I'm not sure if that correlates to that rarity of its practical usage.

Can the formation of drugs against coronavirus also be useful against the Influenzas virus or HIV? This is because these viruses shared many common proteases for viral replication.

Kindly share any research outcomes related to the above topic or virus /influenza based on the contemporary situation.

Antibiotic resistance and the emergence of many new viral strains due to climate change, animal farming and deforestation are increasing the usefulness of pharmaceutical agents, incl. antibiotics. We are currently dealing with yet another viral pandemic. Are there any natural plant extracts that are being studied that are effective disinfectants for the whole suite of fungal, viral and bacterial infections that act as pathogens to humans ? Particularly interested in those effective against coronaviridae ?

Or not?

In The Great Influenza by John Barry at p. 340, about government controlling fear, writes: "They could not control it because every true report had been diluted with lies." At page 460, he writes: "For if there is a single dominant lesson from 1918, it's that governments need to tell the truth in a crisis. Risk communication implies managing the truth. You don't manage the truth. You tell the truth."

An article touching on these issues is, The Only People Panicking Are the People in Charge, by Malka Older, September 16, 2020.

Statistics, lies and the virus: Tim Harford's five lessons from a pandemic, Sep 10, 2020 Financial Times magazine, includes: "Carefully gathering the data we need, analysing it openly and truthfully, ... this is the only chance we have to defeat the virus ...."

I am looking for your opine

I am looking for protocols and/or information as to how to perform sucrose gradient ultracentrifugation to purify samples of influenza A that have been propagated in egg allantoic fluid. Does anyone have any protocols that includes the speeds used? (Protocols that do not cause lysis of the virus after purification are greatly appreciated as well)

Does anyone know what the immunogenic similarities of the two mentioned viruses are (I mean the specialized similarities needed to make a vaccine, such as a specific genomic sequence)?Please give me more information.

Thanks.

UVGI technology has long been known to be an effective tool to fight against airborne infectious viruses. However, lower wavelength UV light source (<240 nm) is also known to induce Ozone production from oxygen in the air. In addition, UV light is known to have carcinogenic and cataractogenic effects. UV lamps of wavelengths in far-UVC range (207 to 222 nm) and low dosage (2 mJ/sq.cm) have been studied to have an effective germicidal effect by neutralizing viruses and bacteria without adverse health effects to humans. Far UVC has emerged as a promising technology in the recent years ( https://www.nature.com/articles/s41598-018-21058-w ). While the use of wavelengths higher than 240 nm can be incorporated in enclosed systems like air purifiers and HVAC which can minimize direct skin and eye exposure of humans to UV light, far UVC has additional advantages of usability in open configuration in public places.

What are some other known real-life challenges and other important issues associated with UVGI technology to be cared for while designing systems integrating HEPA/ULPA filtration to be used in portable air purifiers, HVAC systems or upper room UVGI lamps in hospitals, commercial and residential buildings specially in the context of fighting the rapid spread of infectious viruses like the coronavirus associated with COVID-19?

Does anyone know of a panel of serum/plasma with antibodies to known pathogens such as Influenza A, HBV, HCB etc, that could be purchased for testing? or through ethics application to biobank?

Inhalation of steam from the decoction of mint and oregano can be effective in treating lung infections and coughs caused by COVID-19 or influenza, especially during the recovery period. This has been explored in a very small statistical community.

These plants can act as inhaled antibiotics and can be applied directly to the infected areas, especially the inactive parts of the lungs, and can be effective in treating the infection.

These herbs also have anti-inflammatory properties and can be effective in treating cough and easier breathing of patients.

The NS1 allows that patients asymptomatic can infect.

Hi everyone,

We are synthesizing a virucidal material that will kill SARS-CoV-2 on contact.

In these preliminary stages, we want to use enveloped surrogate viruses that are the easiest to propagate and safe to handle. For example, Phi6 bacteriophage and its easily cultured host, Pseudomonas syringae.

Other CoVs (299e, OC43, etc.) and influenzas have been considered, but as I said for now we want to keep complications to a minimum.

Any advice on possible enveloped viruses to use?

Thanks,

Max Bueckert

COVID-19 is a worldwide threat to humanity and spread fast. There is no effective vaccine yet, the combined antiviral therapies are performed and have a higher success rate. But we need additional treatments and approaches to this disease.1

I and my colleague realized that the levels of high-density lipoprotein (HDL) and non-HDL were very important for sepsis fatality in a small observational study. We tried to predict mortality with lipoprotein levels and ratios. We have believed that there is a balance between HDL and non-HDL, and it is an immunological actor for infectious disease.2 Our study was about bacterial infections, but a similar relationship may exist in viral infections. Hepatitis-C infections may alter lipoprotein metabolism. Statins have used for adjuvant treatment for chronic hepatitis-C infections.3 HDL may be an innate immunity actor, modulate macrophage functions and inhibit thrombotic events. LDL decrease was shown with inflammation.4

A meta-analysis showed that total cholesterol and low-density lipoprotein (LDL) levels were good predictors for Dengue virus severity. The investigators thought the lipoprotein levels should be screened routinely and the approach should be reconsidered with these levels.5

Another study showed that statin users with Influenza had fewer complications and 30-day mortality than non-users. But this protective association was demonstrated for some seasons and subtypes.6

The lipoprotein levels and ratios may be explored in COVID-19 screening and follow-up may be a predictor for prognosis. These levels and ratios may be targets for treatment.

1. Yan Y, Shin WI, Pang YX et al. The First 75 Days of Novel Coronavirus (SARS-CoV-2) Outbreak: Recent Advances, Prevention, and Treatment. Int J Environ Res Public Health. 2020 Mar 30;17(7). pii: E2323. doi: 10.3390/ijerph17072323.

2. Karahan I, Cifci A. Are Lipoprotein Levels and Ratios Able to Predict Mortality due to Sepsis? J Coll Physicians Surg Pak. 2020;30:272-275. doi: 10.29271/jcpsp.2020.03.272.

3. Aizawa Y, Seki N, Nagano T, Abe H. Chronic hepatitis C virus infection and lipoprotein metabolism. World J Gastroenterol. 2015;21:10299-313. doi: 10.3748/wjg.v21.i36.10299.

4. Levine DM, Parker TS, Donnelly TM, Walsh A, Rubin AL. In vivo protection against endotoxin by plasma high density lipoprotein. Proc Natl Acad Sci USA 1993; 90:12040-4.

5. Lima WG, Souza NA, Fernandes SOA, Cardoso VN, Godói IP. Serum lipid profile as a predictor of dengue severity: A systematic review and meta-analysis.

Rev Med Virol. 2019;29:e2056. doi: 10.1002/rmv.2056. Epub 2019 Jun 6.

6. Atamna A, Babitch T, Bracha M Statins and outcomes of hospitalized patients with laboratory-confirmed 2017-2018 influenza. Eur J Clin Microbiol Infect Dis. 2019;38:2341-2348. doi: 10.1007/s10096-019-03684-y.

JAMA article published today states that SARSCOV2 is at least 40 times as deadly as the Influenza virus. Why are most people not concerned? Do they not know this information? Do they think that they are immune? Do they not trust the Health authorities? Do they know that the data, assumptions made, results and conclusions are somewhat suspect of comparing "apples to oranges?" Do they not see evidence to support the studies? I went out today, it seems like I was the only person taking precautions. Why?

Due to their small size, viruses were shown to dry and die quickly, especially on porous (hydrophobic) surfaces and in dry environments.

Could drying the air be a method of treatment / prevention for colds and influenza?

In this review I will elabotely study the complete spectrum of antiviral compounds used in different Viral Diseses - applied to the latest Covid19 Pandemic .The review is mainly based on to point out the Older to Newer antiviral medicine -from Medicinal Synthetic background .

Your openion will be cordially welcome - Thanks

Jaydip Datta

I am interested in measuring the underlying inflammation associated with viral infections (corona virus or influenza) to test the hypothesis that a inflammation-associated severity index might have clinical utility in treating patients. The prototype can be seen at: https://cslide-us.ctimeetingtech.com/actrims/attendee/eposter/poster/66

Is the absence of decision by UK and NL to enforce social distanciation against Covid19 (unlike China, Italy, France, Spain, Germany, USA, etc) caused by memory loss of the positive effects of proactive enforcement of social distancing by US cities in the 1918 influenza pandemic? Death rates were reduced by 50%, source:

Which markers are suitable for assessing the effect of influenza on the heart (FABP or NTproBNP, ELISA)?

The 1918 H1N1 Pandemic Influenza did not have a vaccine. In fact, physicians and scientists didn’t even know what a virus was or how it worked. They knew it was not bacteria and that it did not seem to be alive, but that was about it. But for the transmission, most scientists thought it was a chemical agent of some sort.

Given the scale of this pandemic, it is possible that 2020 is just the first (and worst) year of an annual coronavirus outbreak, similar to how we have an annual flu season.

Coronavirus mutates at a faster rate than influenza (Nextstrain was researching flu when the pandemic started and they are currently tracking mutations in SARS-CoV-2 too). By the time billions of people have been infected, SARS-CoV-2 (the novel coronavirus) may have mutated into a few distinct strains, different enough to each require its own vaccine.

It is also possible that a strain appears that is far less lethal. If this disease was killing 1 in 10,000 instead of 1–5 in 100, we probably wouldn’t even have noticed it until tens of millions had been infected and we’d probably just let it run its course without any actions, just as we do every year with the other coronaviruses that cause the common cold.

As we know, Australian researchers have found that body's immune response to COVID-19 is similar to influenza. Now my question is: "Does getting influenza give some immunity against COVID-19?"

I am interested in the robustness of this technique. I have seen so far publications only from the lab of founders of the technology. Therefore, my curiosity if it is used in the field of vaccine development too

Referring to research papers and general advice are welcomed as part of the discussion

Which markers are suitable for assessing the effect of influenza on the heart (FABP or NTproBNP, ELISA)?

Can either rhinovirus or Dengue virus, for instance, coexist with coronavirus in the same infect cell?

It is know that multiple viruses can coexist in the organism, like HIV and C hepatitis, B and C hepatitis, syncytial respiratory and influenza viruses, Dengue and Chikungunya viruses, etc.

SARS-Cov, SARS-Cov-2, rhinovirus, Dengue virus, among others viruses, are Group IV RNA viruses that affect the respiratory tract, and could target to some extent the same epithelial cells. Peace and blessings.

We want to study about influenza viruses and producing vaccines. In vaccines, 15 mcg HA / 0.5 ml units are determined for each strain. Which methods are used to calculate this concentration? I am glad if you give me information.

Thank you for any advice.

Regards,

1918 influenza pandemic caused death more than that of combined death in World war I & II. Since then world has faced four Influenza pandemics. last one in 2009. We are in threat of more Influenza pandemics. What measures could be taken to prevent the development of Influenza pandemics?

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Hey there,

I've been trying to purify these influenza samples, however, I've come across 2 problems. I've repeated the procedures twice and have come across the same issues both times (first time thinking some error had occurred).

Sample 1: After PCR, product is visualised on gel to see if there is a correct product size. There is but also some non specifics, so I cut the band out. After gel purification, the band is now larger bp than original. Size of PCR product is 916bp. However, after purification, it is now well above 1000 bp.

Sample 2: After gel purification, the product has disappeared.

I know it is not an error of the kits (i.e forgetting to add ethanol to wash buffer or forgetting to incubate) as I have used these same kits to purify many samples before it. So these are perhaps sample-specific problems.

I have run these samples using both the Qiagen Gel Purification kit and the Intron Purification kit. Both times have encountered the same problem.

Any recommendations on what I should try?

Thank you so much!

Hello everyone;

I'm preparing my master thesis it's about influenza prediction using deep learning. The data that I had is the rate of dangerous cases and the suspicious ones per week and per region around the country.

for now I implemented the Random Forest, KNN, DNN, LSTM, CNN, CNN-LSTM and Deep Belife Network. I concluded that I'm faced with time series forecasting so I used the window method to make my problem surpervised with window_size=3, 2 and 1. Calculating the r2_score I got 10 regions under the 70% (which I had read that it's the acceptable threshold).

So I'm writing this question hoping to find a solution or get some idea or another deep learning technique and maybe special architecture of the technique used above to improve my prediction in this region.

and thank you in advance

(You will found attached some picture of the regions that I want to improve and my LSTM model.)

These are not contaminants. I have checked several times. I have used Gibco high glucose DMEM media with 10% Heat inactivated FBS and 1X anti-anti. I have used gibco TrypLE for trypsinization for around 10 mins. I do not understand what it is?

Interested in bio security aspect of disposing of horse faeces in an Equine Influenza 2 outbreak. Cannot find any research regarding if the virus is or is not shed in droppings. Also, how long would the virus remain infectious in the bedding?

Many thanks

Canine influenza viruses (CIV) are contagious respiratory diseases found in dogs. These Type A influenza viruses and the H3N2 (canine and human) virus is known to infect dogs, cats and people. There is a concern that the disease can mutate with dogs being the ”mixing bowl” and potentially produce a pandemic. See: https://www.ingentaconnect.com/search/article?option2=author&value2=Daesub+song&pageSize=10&index=1#

I am attempting to perform a qPCR standard curve using viral DNA (Adenovirus and MCMV) and cDNA (Influenza A) to be used to determine relative abundance of these viruses in primary samples. Viral DNA and RNA was extracted from viral stock using the QIAmp MinElute Virus Spink kit and vRNA was converted to cDNA using the ImPromII Reverse Transcription System. 6 point 10x dilutions were performed using this viral DNA and cDNA (undiluted-1:100,000) and a negative water control was included. For the qPCR, I am using TaqMan Universal qPCR Master Mix along with virus specific primers (forward and reverse) and probes whose sequences were provided from literature review or provided by neighboring labs but have not previously been used by our lab with this particular master mix. I believe that all of the input DNA/cDNA concentrations are fine (ranging from 17-180 ng/rxn note: master mix prefers concentrations <250 ng) and being that I have repeated these experiments a number of times with great care, I do not believe that it was a pipetting error with the dilutions. The amplifications curves, especially for Adenovirus, do not follow the natural curve progression seen in most standard curves and for all of the viruses the points do not fall on the slope of the standard curve line. I have attached photos for reference. I have tried repeating these experiments many times and adjusting the input concentrations of the viral DNA and cDNA. All experiments were for each virus was done separately, there was no multiplexing. I am wondering if it is possible that the primer efficiency may be the problem. The annealing temperatures for these primers are slightly higher (~3-6 degrees) than recommended by the master mix but a representative from Fisher seemed to believe that the reaction should still work. Also the target regions for these primers seem very short (all <150nt). Is it possible that I need to design new primers or is there possibly another obvious issue. If anyone could provide any feedback on what they believe the possible problem is or has further questions, please let me know.

I m using dh5 alpha cells to insert some specific segments from influenza A . I ligated the cells using TA cloning kit and transform them . I cultivated the cells on an LB media with ampicilin , Xgal and IPTG for the blue white screening. The screening worked perfectly , I cultivated them overnight and after this I used the Qiagen mini prep kit. I used ecor1 to digest the mini prep and after this i run a gel electrophoresis (agrose at 1%) for 45 minutes at 70V . I can't understand why I got the top bands like this , is it super coiled DNA ? Is something normal or I didn't do any of the steps properly . Does somebody know where I failed ?

Hello,

I was doing a PCR for influenza viruses ( more specifically defective interfering segments) and i had good results beside 2 segments. I store them at -20 degrees and i was thinking to run them 5 cycles more ( initially all the sample were run for 15 cycles) . Do i need to make fresh samples or i can just reuse the 2 stored samples. I'm thinking just to place them in the PCR machine and add 5 more cycles to this 2 samples .

Will this work ?

I know the buffer, dNTPs , Taq polymerase and others are limited and have a specific life expectancy of PCR cycles, but 20 ( 15 already done + 5 which i will run )cycles from my presumption will not exhaust all the dNTPs and other reagents.

Thanks a lot ! Sorry for this stupid question !

Hi,

I want to see if my mice have gotten infected, is it possible to coat an ELISA plate with IAV broken up by sonication?

Thank you!

Hi, I want a correct protocol for plaque assay with influenza viruses .

I used MDCK CELLS ( 450 000 cell/well ) with different strains and different plates but i didn't see plaque formation but only holes on all conditions ( also control).

What do you think about this problem?

Can i have a correct protocol of it?

thank you

best regards

Vitamin D status is associated with upper respiratory tract infection and / or influenza.

Of course such association (or causal relation) is stronger when there is :

- initially state of deficiency (lower than 50 nmol/L or 20 mg/nl)

- sufficient high dose of suppletion (e.g. 1000 - 2000 iU / d)

- time of suppletion is long enough (at least 3 month) before the season of cold / influenza etc starts

I point at a metaanalysis of Martineau et al (BMJ 2017) and know of some special RCT's but I would like to discuss this with you and maybe find some extra evidence regarding a positive association / causal relation.

Martineau Vitamin D supplementation to prevent acute respiratory tract infections: systematic review and meta-analysis of individual participant data

BMJ 2017;356:i6583 | doi: 10.1136/bmj.i6583

Poultry diseases cause increased mortality and high economic losses. Is it possible to breed to produce disease-resistant poultry strains?

How to seek for the UTR of IAV?

Is there any database to seek for the UTR of influenza?

I need to know the UTR sequence of influenza to rescue IAV.

I was using MDCK cells to grow Influenza H1N1 in 12-wells plates. Cells was seeded 1 x 10^6 cells/well and left overnight in media without serum. Cells were then infected with H1N1 at moi of 0.1 and virus titre was tested by HA assay after 24 hrs, and 48 hrs.

Trypsin was added during infection.

Under the microscope, the cells was achieved ~80% cytopathic effect after 48 hours. However, no titre was determined by HA assay (all negative HA) either after 24 hours or 48 hours.

Is there any mistake happening here?

In my point of view, first, I guess its because of the temperature during harvesting, virus was harvested at room temperature from 37C cell culture and spin at 4C and then store at -80C freezer for further HA testing.

Second point, was the virus titre too low? or was the cells density too low?

Does anyone here working in the same field or experiment with me before?

I'm currently performing the MN assay using A/Hong Kong/4801/2014 (H3N2) strain of influenza. I came across a strange change in cell layer post 3 days of inoculation with the virus. The cells were detached from the surface of the well and formed clumps. Surprisingly, the clumps are found around the corners, so it has generated half-moon shape in the well where half of the cells are attached smoothly and rest half are detached and clumped.

I seeded cells 2 days ago and then used the plates for the assay.

Thank you in advance.

The viral RNA obtained from a patient with influenza symptoms was analyzed and the results for a real-time PCR were negative for the influenza A virus and positive for the AH1NI virus.

The positive and negative control were fine.