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Wuhan Coronavirus: NATO economic weapon? China virology lab?
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TonyGosling
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PostPosted: Sun Jun 14, 2020 11:00 am    Post subject: Reply with quote

DEVASTATING
NHS say only 3300 deaths where there are no underlying conditions and Covid19 is the only cause of death

Early June graphs from Christopher Bowyer
https://hectordrummond.com/2020/06/05/early-june-graphs-from-christoph er-bowyer/

POSTED FRIDAY 5TH JUNE 2020 HECTOR DRUMMOND

Reader Christopher Bowyer has once again at the digital coalface making graphs, which he has sent to me. Some are based on NHS England data, and some on the ONS week 21 release.

(Bear in mind that ‘with Covid-19’ deaths includes any death where the deceased was suspected of having Covid-19, even if it was not thought to be the cause of death, so these numbers are probably inflated.)



First, NHS England deaths with Covid-19 by absence (yellow) or presence (green) of a pre-existing condition.





Next, percentage of deaths with Covid-19 by pre-existing condition. (95.13% have one or more P.E.C.) Data from NHS England.



Note that the most common pre-existing conditions are diabetes, followed by dementia, chronic pulmonary disease, and chronic kidney disease.





Covid-19 deaths in England by date of death, with 3-day moving average trendline (the thicker line). (Note that numbers in grey area will increase over the next few days.)





A graph of deaths with Covid-19 in England which occurred each day, by the delay in reporting. Thick line is overall daily announced deaths.





Trendlines (3-day average) for Covid-19 deaths by England NHS region, by date of death. (Note that the numbers in the grey area will increase over the next few days.)





Trendlines (3-day average) for Covid-19 deaths per million by England NHS region, by date of death. (Note that the numbers in the grey area will increase over the next few days.)





English daily deaths with Covid-19 by date of death. Blue line is all Covid deaths, red line is hospital Covid deaths, and green line is non-hospital Covid deaths.





Weekly cumulative deaths with COVID-19, by age range (ONS data).





Total deaths with COVID-19, by detailed age range (ONS data).





Care home deaths for England and Wales. Five-year average deaths are blue, Covid-19 deaths are red, and excess non-Covid deaths are green.





Weekly Covid and non-Covid deaths at home. Five-year average deaths are blue, Covid-19 deaths are red, and excess non-Covid deaths are green.





Weekly Covid and non-Covid deaths by place of death. ONS data.





Many thanks to Christopher for doing these once again.

Update: Please help out with donations or a Patreon sub if you can, or at least buy my book! This is just the start, but I need your support.



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_________________
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www.mp911truth.org
www.ae911truth.org
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www.v911t.org
www.thisweek.org.uk
www.abolishwar.org.uk
www.elementary.org.uk
www.radio4all.net/index.php/contributor/2149
http://utangente.free.fr/2003/media2003.pdf
"The maintenance of secrets acts like a psychic poison which alienates the possessor from the community" Carl Jung
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TonyGosling
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PostPosted: Sun Jun 14, 2020 2:04 pm    Post subject: Reply with quote

Veater Ecosan: "The World About Us"
https://veaterecosan.blogspot.com/2020/06/a-step-by-step-guide-to-worr ying-world.html
"Plebs omnis plaudit ut me minore sepius audit." TREBLE BELL, COMBE RALEIGH CHURCH, DEVON.

Friday, 12 June 2020
A 'STEP BY STEP' GUIDE TO WORRYING WORLD-WIDE DEVELOPMENTS

View image on Twitter
There is no end to the predictive programming. This one is 'bat-* crazy': The reverse side of the 25-cent coin depicts a Samoan fruit bat mother with her pup clinging to her wings released by the US Mint on Feb. 3, 2020.


Authored by a member of a group that wants to remain anonymous. The unabridged article can be viewed here:
http://www.marcvandelft.nl/corona-crisis/Manual.pdf?fbclid=IwAR32RLpIS 4GiOLAZ9Omu_L4mdMx5qrhQfHU-75osNJBmU37R3cArw-VaWcU

The following is a 'step by step' guide to what we believe are the worrying international developments, the objective of which is to spread awareness in the general public, crucial if the dangers described are to be frustrated.

https://l.facebook.com/l.php?u=http%3A%2F%2Fwww.marcvandelft.nl%2Fcoro na-crisis%2FManual.pdf%3Ffbclid%3DIwAR1VyAxWGx7Kia-RBL48Ca3JtslA4JCF60 DOr0IjeEH2kisTBZ7cT_X9EFk&h=AT3ssV_SDv90WNbikGfKwUs28JHFcpNE49-wESam-d CwS6zK1ys8gkRAn3rTshD1ebL_6ScHKJiNfbJf3MehOH6EkOl1jB1tAcqixiOXFhkgn23G fNZiGUL_ecrApbf5v08

We believe what is now unfolding can be compared to Thomas Mann's brilliant novel 'Der Zauberberg'. The hero of it, Hans Castorp, suffering from TB, resides in the comfortable bubble of a mountain health resort, isolated from the outside world. When he recovers from the illness, he returns to it, only to discover Europe is ravaged by World War I, where he will most likely die. We believe that unless people choose to leave their 'bubbles', they may wake up to a society far more dangerous and dystopian.


There appears to be a covert agenda behind the government measures all around the globe, and our estimation is that, as time progresses, more and more of this agenda will become clear to ever more people. None of us can now afford the luxury of remaining in the bubble of our comfort zones.


As a group, we have known about the most horrendous government schemes for more than ten years now, and we feel it is our moral duty to inform you. In this manual, we take one step at a time. It isn’t our aim to convince you of anything, but simply to present facts in a certain order, so you can then form you own opinion on the basis of these facts.

TWENTY STEPS TO AWARENESS



INTRODUCTION

We are now living in a crucial moment in the history of mankind. It is of vital importance to have an understanding of what is now unfolding in order to be able to influence the outcome. Fortunately, the number of people who are aware is growing but needs to expand if disastrous consequences to humanity are to be avoided in the next few years. Hopefully this document will inspire others to do likewise.

We have found information suggesting that there are reasons behind government measures and that unchallenged, a covert plan is being wheeled out on an unsuspecting public. We have witnessed the formation of multiple separate groups that, although having insight, are insular, so that general awareness is limited, which we are trying here to correct.

People differ in their interpretations of what is needed. Some see Donald Trump as a hero, another sees him as a villain. We take the view that it doesn't really matter, because it only obscures the more important issue. Contentious issues such as ‘chem-trails’, the ‘Illuminati’, 9-11, ‘Qanon’, are similarly secondary. This is about joining forces on the basis of common ground. People need to subsume their differences in order to unite against the common enemy.


CORONAVIRUS CASE STUDY


At least deep down, everyone will have a feeling that there is something strange about the uniform measures that many governments have taken worldwide.

locking all of us up,
crashing the economy,
causing damage to many isolated elderly people (83-year father would rather die than continue to live in corona quarantine): https://www.youtube.com/watch?v=efmEfeopqfE&t=4s)
damaging our immune systems by ordering us to stay inside
feeding us with enormous fear,
splitting grieving families,
preventing normal funerals,
the dying have been forced to say their farewells via iPads,
while on the other hand prisoners are released everywhere ‘to combat the virus’ (https://www.foxnews.com/health/us-starts-release-inmates-coronavirus, https://www.reuters.com/article/us-health-coronavirus-iran-idUSKBN20W1 E5, etc.).

Another ‘side effect’ of the government measures is that the number of suicides has risen dramatically: https://abc7news.com/suicide-covid-19-coronavirus-rates-during-pandemi c-death-by/6201962/.

At the same time, more and more information is appearing indicating that the mortality rate of the current virus is no higher than that of an ordinary flu (https://californiaglobe.com/section-2/coronavirushysteria-the-numbers -dont-warrant-the-media-hype/).

Isn’t it striking, too, that none of the government advice deals with things like healthy nutrition, vitamins such a 'C and D, enough exercise, doing things that bring joy, etc., known to reinforce the essential human immune system? And isn’t it strange that such news doesn’t appear in the mainstream media?

Obviously, all of the above-mentioned measures have considerable negative side effects. It is only logical that these could have been predicted and planned for, but despite detailed legislation, the general impression was one of panic and chaos.

People will recall the attacks of 9/11 were said by leading politicians to have been 'completely unforeseeable' and could not have been predicted, despite previous attacks of a similar kind that exercises had anticipated. In similar vein, governments have demonstrated unpreparedness (absence of PPE, testing equipment, immigrant testing and control etc) for the Coronavirus outbreak, despite clear evidence of pre-dating anticipation.

Some examples of the specific expectation of a Coronavirus pandemic follow. The content and timing of these events and documents either display extraordinary prescience, or god forbid the sort of planning with numerous other so-called unexpected 'terrorist events'.

A. In the 2018 report Road Map for the Implementation of Actions by the European Commission announced that the project for mandatory vaccination of each and every world citizen is to be completed by 2022, See: https://ec.europa.eu/health/sites/health/files/vaccination/docs/2019-2 022_roadmap_en.pdf

B. The Global Preparedness Monitoring Board (GPMB) 2019 Annual Report: 'A world at Risk', strongly hints at a second wave, See: https://apps.who.int/gpmb/assets/annual_report/GPMB_annualreport_2019. pdf utm_source=mandiner&utm_medium=link&utm_campaign=mandiner_202004.
The Global Preparedness Monitoring Board (GPMB), a new body set up under the aegis of the WHO and the World Bank Group to monitor the world’s readiness to respond to outbreaks and other health emergencies with the financial support provided by the Government of Germany, the Bill and Melinda Gates Foundation, the Wellcome Trust, and Resolve to Save Lives.". It had has its origins in the 2014 Ebola outbreak in West Africa but convened the first meeting on 10th Sept, 2018. The board aims to publish its first report on the global state of preparedness in September 2019. https://www.who.int/news-room/detail/10-09-2018-global-preparedness-mo nitoring-board-convenes-for-the-first-time-in-geneva The timing could hardly be more propitious but not quite 9/11!

C. Event 201 was a 3.5-hour pandemic tabletop exercise that simulated response to a hypothetical, but scientifically plausible, pandemic.Similar to the Center’s 3 previous exercises—Clade X, Dark Winter, and Atlantic Storm—Event 201 aimed to educate senior leaders at the highest level of US and international governments and leaders in global industries.

"Fifteen global business, government, and public health leaders were players in the simulation exercise that highlighted unresolved real-world policy and economic issues that could be solved with sufficient political will, financial investment, and attention now and in the future."

"The next severe pandemic will not only cause great illness and loss of life but could also trigger major cascading economic and societal consequences that could contribute greatly to global impact and suffering. The Event 201 pandemic exercise, conducted on October 18, 2019, vividly demonstrated a number of these important gaps in pandemic preparedness as well as some of the elements of the solutions between the public and private sectors that will be needed to fill them. The Johns Hopkins Center for Health Security, World Economic Forum, and Bill & Melinda Gates Foundation organised the event and jointly proposed these recommendations."

It cannot escape notice how the Gates Foundation keeps cropping up whenever pandemic or vaccination programmes are considered, not to mention tracking technology. Imperial College London, whose Ferguson models predicted 500,000 UK deaths, had apparently been the beneficiary of over £200 million in Gates Foundation grants.


DEVELOPMENTS IN HOLLAND



They are not normal, and shouldn't be in a democratic society either ". According to the Order, emergency regulations provide plenty of space and a new law is not necessary. (https://bit.ly/nw-adviesnova
)

And the College of Human Rights also delivered an opinion: https://bit.ly/nw-mronline

These advice are valuable but not binding during the approval process.
𝗩𝗥𝗔𝗔𝗚𝗧𝗘𝗞𝗘𝗡𝗦
The bill brings a lot of questions. Submission comes at a time when the figures of the RIVM look positive: the number of hospital recordings and corona deaths has decreased drastically in recent weeks and is steadily heading towards the zero line. See the report ′′ Epidemiological situation COVID-19 in the Netherlands 5 June 2020 ": https://bit.ly/nw-rivm-rapport
.

Of course it is in the public interest to ensure that numbers do not rise drastically again. But there are other, less far-reaching possibilities for this, which do not affect our freedom, fundamental rights and human values.
I wonder:
✨ Why at the moment many rules, punishments and extensive powers for the government in the law?
✨ Why a 1,5 meter rule in the law, also for outside? (https://bit.ly/nw-buiten
, https://bit.ly/nw-smartexit
)
✨ Why the high fines?
✨ Why the possibility of prison and community service?
✨ Why now the possibility of breaching our domestic right?

It would be expected that there will be less heavy rules now than at the beginning of the crisis, but nothing is less true.
Furthermore, the law would be in the interest of public health. If it really is:
✨ How is it that the new rules have undeniable negative consequences for our health? (https://bit.ly/nw-artsen
, https://bit.ly/nw-contact-levensbehoefte
)
✨ Then why is it allowed that the law will cause huge economic and social damage, which also has major consequences for our wellbeing and our health? (https://bit.ly/nw-cbs
, https://bit.ly/nw-schade-smartexit
, https://bit.ly/nw-ira
)
✨ Why is not (also) deployed to improve our resistance and is the advice of 1600 care professional not taken seriously? (https://bit.ly/nw-leefstijl
, https://bit.ly/nw-zelfzorg
)

I've always learned that the means must sacred the end. A mosquito won't kill you with a rocket launcher. When I look at the figures of the RIVM and the intended purpose of the law, I see no reason to use these far-reaching, freedom restrictive means and understand the critical attitude of the advisory parties (see above).
I also find it incomprehensible that this law was not discussed during the press conference of Wednesday 5 June. There was mainly talk about ′′ the large package of relaxation ", as Rutte calls it, not about this emergency law, which is a large rule and penalty increase for an indefinite period.
𝗩𝗜𝗝𝗙 𝗩𝗢𝗢𝗥 𝗧𝗪𝗔𝗔𝗟𝗙 𝗘𝗡 𝗪𝗔𝗧 𝗞𝗔𝗡 Í𝗞 𝗗𝗢𝗘𝗡?
As far as I'm concerned, everyone should be aware of the consequences of this bill. We can then take joint action to prevent this proposal from being approved. Fortunately, the newspaper and journal have picked up the subject, but it's still five to twelve.
After the (non-binding) advice of the Council of State, only the First and Second Chamber can stop the law. We can help with that by letting us know:
✅ Sign the main petitions: https://bit.ly/nw-smartexit
, https://bit.ly/nw-petitie
, and https://bit.ly/nw-petitie-vw

✅ Join the Facebook group ′′ NO against 1,5 meters ": https://bit.ly/nw-fb-nee
, at https://www.1.5xbeter.nu
and https://viruswaanzin.nl

✅ Share this post with as many people as possible or write a piece about it yourself
✅ Inquire your friends and neighbors

If you have any more ideas to spread this news or stop the law approval, please share it with mi
Mr. Online (https://bit.ly/nw-mronline
)
Criticism of proposal corona law - Mr. Online
The Dutch Order of Lawyers is critical of the legislative proposal Temporary law measures COVID-19. According to the College of Human Rights, ' human rights risk s' are attached to the law. The law is designed to create a legal basis for corona measures in the other half meter society for a longer term. These are now based on emergency regulations. Legislation [...]

_________________
www.lawyerscommitteefor9-11inquiry.org
www.rethink911.org
www.patriotsquestion911.com
www.actorsandartistsfor911truth.org
www.mediafor911truth.org
www.pilotsfor911truth.org
www.mp911truth.org
www.ae911truth.org
www.rl911truth.org
www.stj911.org
www.v911t.org
www.thisweek.org.uk
www.abolishwar.org.uk
www.elementary.org.uk
www.radio4all.net/index.php/contributor/2149
http://utangente.free.fr/2003/media2003.pdf
"The maintenance of secrets acts like a psychic poison which alienates the possessor from the community" Carl Jung
https://37.220.108.147/members/www.bilderberg.org/phpBB2/
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TonyGosling
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Location: St. Pauls, Bristol, England

PostPosted: Fri Jun 26, 2020 12:48 am    Post subject: Reply with quote

Never Again
June 22, 2020
https://johnplatinumgoss.com/2020/06/22/never-again/

Never in my fairly long life have I been exposed to such a global scam as the way in which the virus SARS-COV-2 has been used to create public panic and distrust in the world. Humans are naturally gregarious animals yet in practically every country on the planet they have been instructed to disobey their friendly and communal instincts in favour of “social-distancing”. Stay two metres apart has been the general message.

Nobody knows exactly how viruses are spread or exactly how people become infected. Nevertheless, there has been a universal message – almost a demand – to keep well removed from our neighbours and other people outside of our immediate families. Two metres was the original instruction, a measure which caused lengthy queues at food markets and elsewhere, brought industry to its knees and sent shivers through the stock markets. More recently this distance has been reduced to one metre in some countries and that may well become, “the new norm” in the UK from 4 July. That suddenly the distance can be halved demonstrates that the gurus who put out these messages have absolutely no idea what distance is a safe distance, if any.

For three months the message from mainstream media was repetitive and preclusive of any other argument. Only recently has there been any questioning and then precious little – outside of social media articles which have been quick to query what has happened. Since it became quite clear that SARS-COV-2 was simply another virus – not dissimilar from AIDS, Ebola, influenza, the common cold or shingles – and like those viruses would ultimately run its course without wiping out the entire population, there has been a mellowing of the perpetual indoctrination which was tedious.

Thanks to thinking people, who have suffered much abuse by questioning the narrative, even its perpetrators can now see that the plan has not worked. The way the corporate media have presented a panic which they themselves created has been anything but balanced. What’s more that presentation took the form of a series of imperatives penetrating our eardrums and brains with instructions accompanied by “you must”. Visually this has been further endorsed by newsreaders sitting well apart from one another, most of the time looking awkward and ungainly.

This raises many questions. Who has the power to control the global media and governments? Who can get practically every country to sing the same song with the same orchestral backing, while its real entertainers and theatrical productions are sidelined? Who can bring all sporting events played by and within nations to a standstill? Why would they want to do this?

The power wielders
Most people, at least for a month or two, have been happy to comply with “the new norm” created for them by entities outside of their personal networks. Out of these dedicated followers of mainstream opinion there are those who tell anyone who prefers to question the narrative that it would not be possible to get all the countries and their governments to go along with a hoax. Is that true?

On the face of things it does seem doubtful that all countries would comply. Who could possibly get the people portrayed as ideological enemies of the west, like Xi Jinping and Vladimir Putin, to adopt this global plan? Originally Russia presented very few coronavirus infection figures with no deaths making it look like closing its borders had worked. Suddenly those figures changed quite dramatically and Russia became one of the worst hit countries, unlike its neighbour, Belarus (more of which later).

History can teach lessons. In the nineteenth century the banker Nathan Mayer Rothschild said: “The man who controls Britain’s money supply controls the British Empire, and I control the British money supply.” In this honest self appraisal lies all the wisdom necessary to see what has happened recently with the coronavirus panic-demic. He who pays the piper calls the tune. And that answers one of the big questions raised. The panic caused by a virus, less deadly by far than HIV-Aids, was yet another attempt by the elite to gain total control of the planet. The only real difference is that this most recent imperialistic excursion into all countries simultaneously was attempted without resorting to war, which on the face of it seems less cruel.

In the early 21st century a Rockefeller descendant, Nathan Rockefeller, told Aaron Russo what the elite families’ blueprint was for controlling and enslaving ordinary people and changing the world forever. Film producer, Russo, who died of bladder cancer in 2007, was not happy with what he heard and decided to go public. In an interview with Alex Jones he talked about how 9/11 had been planned to create a new enemy out of Islamic nations, how feminism had been introduced to create a rift between man and woman together with other control mechanisms the elite were using, and had planned, for global supremacy through dividing and conquering. Nathan Rockefeller told Russo how the world’s public, that’s you and me, would be micro-chipped to monitor our spending preferences and financial transactions and to make sure we did not step out of line – in other words modern slavery. This two minute extract should be enough to scare the pants off any thinking person.

Panic pandemonium
The death figures presented deliberately to create panic are not as scary as the way they are presented. As well as the argument whether a person has died “from” coronavirus or “with” coronavirus, global death-rates are pretty much on a par with other years and any excess of deaths may well be accounted for by lack of medical care for those on waiting-lists due to the extra resources ploughed into treating COVID-19 cases. The eventual figures will bring the truth to light.

Country to country comparisons have been used to question why a neighbouring country might not have been affected as badly as one’s own. All kinds of speculation go into the causes for this anomaly. Of the Scandinavian countries Sweden has been exemplified and pilloried as the way not to go. There was no lockdown in Sweden and the pundits who support the “panic pandemonium” have seized on this to highlight Sweden’s death rates in comparison with Norway and Denmark, countries which have had fewer deaths per capita. So is Sweden experiencing a worse than average death-rate? No.

A study from 1990 to 2020 compiled by HaraldofW from SCB (Statistics Sweden, a government agency) shows that death rates from all causes for the months January to April are up on 2019 but down on 2018 and in fact only higher than three other years in that 30 year period.

Deaths per 100 000 Sweden Jan - Apr 1990 - 2020

Belarus is a country which also abjured lockdown measures. It has played all its football matches in three divisions, with normal crowd attendance, and has already started its cup run. Life goes on as usual there. Social interaction has not been curbed and infections from the virus have been high. Deaths however have been very low. There is a lot to be said about the importance of getting out into the fresh air to fight any virus and being cooped up in a claustrophobic environment with the demand to “stay home” and breathe in your own carbon dioxide is an unhealthy message.

While mainstream media have put the wind up everybody there is absolutely nothing more to worry about than there is from the annual influenza toll, less in fact. For some unaccountable reason this virus took on a guise of being some kind of super-killer that could fly around like a caped crusader. That was a big lie. This graph shows the biggest daily killers and Covid-19 is way down the list.

deaths worldwide all causes

Since you are nearly five times more likely to die of measles why have we not had lockdowns to protect us from measles and those with it? Since you are nearly 40 times more likely to die of HIV-AIDS why have we not got social distancing measures over that? Whatever the cause of death it is distressing for families and loved ones at a time when we ought to be giving one another a friendly hug.

For the control reasons outlined above it is imperative that the world wakes up to what is happening and the nonsense that SARS-COV-2 has become for us all. Being brainwashed by a media and governments that are under the control of big bankers has virtually ruined economies of the world and recovering from it will take some time.

This virus has been turned into a monster it should never have become. Accompanying terminology that has grown exponentially with it includes phrases like “the new norm”, “normality will never be the same” together with a whole load of military clichés including “lockdown”. Well thankfully the war is nearly over. While not being a “war to end all wars” it has been a war where all the ammunition and war propaganda has come from one side. The most important lesson we can all learn from this “enemy within” is to realise we have been had and next time to say “never again”.

IMPORTANT UPDATE: People have questioned the second chart in this blog-post and I agree with them. The graph is from early March and deaths in which COVID-19 has been cited as a cause have increased considerably since then, even though they are now on the wane in terms of daily deaths. What mislead me was the sources of the chart: the WHO, Lancet and Centres for Disease Control which seemed authoritative enough. While there may not be more likelihood of dying of measles, according to a new report more than 2.4 million people worldwide are killed annually from HIV, tuberculosis and malaria. Johns Hopkins university shows 486,101 have died from (more likely with) COVID-19 up to today (26/06/2020).

_________________
www.lawyerscommitteefor9-11inquiry.org
www.rethink911.org
www.patriotsquestion911.com
www.actorsandartistsfor911truth.org
www.mediafor911truth.org
www.pilotsfor911truth.org
www.mp911truth.org
www.ae911truth.org
www.rl911truth.org
www.stj911.org
www.v911t.org
www.thisweek.org.uk
www.abolishwar.org.uk
www.elementary.org.uk
www.radio4all.net/index.php/contributor/2149
http://utangente.free.fr/2003/media2003.pdf
"The maintenance of secrets acts like a psychic poison which alienates the possessor from the community" Carl Jung
https://37.220.108.147/members/www.bilderberg.org/phpBB2/
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TonyGosling
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PostPosted: Tue Jun 30, 2020 12:34 pm    Post subject: Reply with quote

Deloitte get the loot
https://morningstaronline.co.uk/article/c/deloitte-get-loot

A drive through testing facility for COVID-19 at Edinburgh Airport
IN ANOTHER example of how the coronavirus crisis has driven privatisation, Deloitte were given full charge of the drive-through Covid-19 testing programme: the government sees a medical emergency and turns to accountants and management consultants. Deloitte subcontracted much of the work to the likes of Sodexo and Mitie.

A new report reveals Deloitte also played a central role in the development of NHS Supply Chain, which has struggled so badly to provide PPE supplies to NHS workers in the pandemic.

Its involvement emerged in a report by University of Greenwich professor David Hall, co-published with the anti-privatisation group We Own It. Dr John Lister, both a health expert and redoubtable campaigner also co-wrote the report, which examines the history of the agency that buys and distributes most NHS “consumables,” including PPE. It’s already had some coverage in the Morning Star, but I wanted to look at some details.

NHS Supply Chain was first formed in 2006 when Labour privatised NHS Logistics. Many hospital trusts continued to buy their own consumables, however, often forming consortia to bypass the privatised, centralised NHS Supply Chain. So in 2017, then-health secretary Jeremy Hunt set up Supply Chain Co-ordination Ltd (SCCL) to transform and manage NHS Supply Chain. Around £500m was taken from trusts to force them to buy more via the revamped central system.

Though state-owned, SCCL is run more like a private firm, managed by Jin Sahota, a former French telecoms executive, helped by Rob Houghton, a former Post Office IT boss whose review into the scandal-hit Horizon computer system was mysteriously abandoned.

Having drawn money in from NHS hospitals, SCCL then passes on both the purchasing and supply to private companies under a series of contracts with firms such as DHL, Vizient, Akeso, Compass and Unipart doing the work under “eleven specialist buying functions, known as category towers.” As Hall’s report makes clear, this baroque system puts hospital money into a complex web of contracted middlemen, often at some remove from actual manufacturers.

The government’s aim might have been to cut hospital purchasing costs, but the system, as seen in the Covid-19 crisis, has failed spectacularly in deliveries of PPE supplies. So who might have designed a system that serves the public sector so badly? Management consultants and accountants. In 2017, Ernst & Young was given up to £20m to design the new central purchasing system. And in 2018, Deloitte was paid £400,000 to design the “category towers” buying system.

Deloitte’s role has angered experienced NHS buyers. The Health Care Supply Association, representing NHS buyers, pointed out that “the NHS has enough capacity and knowledge to be part of the solution, rather than have the solution done to us.” Two years on, NHS Supply Chain’s centralised PPE shortages do resemble something done to rather than by the NHS.

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PostPosted: Sun Jul 05, 2020 11:29 pm    Post subject: Reply with quote

How Dr. Wolfgang Wodarg sees the current Corona pandemic

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https://www.youtube.com/watch?v=p_AyuhbnPOI

How Dr. Wolfgang Wodarg sees the current Corona pandemic
2,370,010 views
Published on 13 Mar 2020
Dr. Wolfgang Wodarg is the first specialist we met to understand the current crisis about the coronavirus. Please support us so that we can investigate further in making a 90min cinema documentary: https://www.indiegogo.com/projects/co...

We will meet multiple personalities with different points of view and we will do extensive fact-based research. During our path to the truth, we think it’s important to open the discussion and analyse all sides. We intend to gather a maximum of information and then help to understand what is happening, and why it is happening.

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Martin Van Creveld: Let me quote General Moshe Dayan: "Israel must be like a mad dog, too dangerous to bother."
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PostPosted: Wed Jul 08, 2020 11:58 pm    Post subject: Reply with quote

'Six months before the pandemic, Bill Gates negotiated a $100 million contact tracing deal with a democratic congressman':
https://www.naturalnews.com/2020-07-08-pandemic-bill-gates-negotiated- contact-tracing-deal.html
'Four months before SARS-CoV-2 began infecting the people of China; Bill Gates was busy negotiating a $100 billion contact tracing program to be implemented by governments and forced on all Americans. On the week of August 12th to 19th, the Gates Foundation met privately with U.S. Congressman Bobby L. Rush (D-IL) in Rwanda, East Africa. The week-long event was underwritten by the Bill and Melinda Gates Foundation and the Rockefeller Brothers Fund.

At the meeting, they discussed the rollout of wide scale contact tracing and negotiated which companies would get to cash in on the plan. They discussed how to contact trace all Americans, how to force them to submit to medical tests and accept vaccination passports in order to go about their lives. (Related: Airline industry pushing for medical testing, thermal screening, digital IDs and vaccination passports.)

Bill Gates was making plans to trace people months before the pandemic was released
The Bill and Melinda Gates Foundation were preparing to test, track, trace, mask, isolate and corner Americans into forced vaccinations four months before SARS-CoV-2 arrived in China and six months before Bill Gate’s friends at the World Health Organization (WHO) declared a worldwide “pandemic.”


Bill Gates is the number one funder of WHO and has been parading media outlets calling for government surveillance of human movement and the need for elaborate contact tracing programs to enforce it. Among these tracing efforts, Bill Gates wants to enforce medical requirements and a “new normal” medical police state, complete with temperature checks that serve as a foothold into forced medical testing and isolation orders. This “new normal” also includes monitoring where people travel and who they interact with, while forcing their contacts into isolation. It all leads to the rollout of vaccination passports that will serve as permission slips for people to gather, meet, travel, and interact. Corporations will be incentivized to participate or be threatened with litigation if they do not go along with a “safe, new normal.”

These shocking revelations were revealed on the Thomas Paine podcast by John Moynihan and Larry Doyle — two undercover investigators who testified in Congress in 2018 about the $2.5 billion tax evasion and fraud schemes employed by the Clintons. Now the investigators are sounding the alarm — how Bill Gates planned to contact trace Americans long before a pandemic was ever declared.

The newly introduced TRACE Act was negotiated long before the “pandemic” was declared
Now, just nine months after the Gates Foundation met with democratic congressman Bobby Rush, there is a bill introduced in the Congress called H.R. 6666, the COVID-19 Testing, Reaching and Contacting Everyone (TRACE) Act. The $100 billion bill was introduced by none other than Bobby L. Rush himself. The TRACE act gives the CDC absolute power over the country and people’s lives, making nasal probes, surveillance tracking, and isolation orders, among other obscene medical edicts, a requirement into the future.


It should be noted that Congressman Rush is from Illinois, a state that is under strict controls put in place from Governor J.B Pritzker. This governor has a financial stake in covid-19 testing companies. This governor also has family connections to Microsoft and Bill Gates and has echoed Bill Gates by saying that the state will not be able to fully open until a vaccine or pharmaceutical is ready to be deployed. The medical coercion in Illinois is very real; Illinois is currently a concentration camp, where citizens, restricted of their liberties, are being conditioned to accept a “new normal” in order to prepare for tracking, tracing, medical requirements everywhere they go, which will likely include coerced, experimental injections by year’s end.'

Just like 9/11, some creatures knew what was coming....

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PostPosted: Thu Jul 09, 2020 12:09 am    Post subject: Reply with quote

'Cover-up: Dr. Anthony Fauci helped approve an effective treatment for coronavirus infections 15 years ago, but is suppressing it today in favor of new high-profit vaccines':
https://www.naturalnews.com/2020-07-08-dr-fauci-approved-treatment-cor onavirus-suppressing-for-vaccines.html
Poor old Faust (sorry, Fauci) wouldn't want people buying tuppenny-ha'penny HCQ when they should buy his expensive new-fangled experimental 'vaccines', now, would he? And if they fail, you can always get 5% off his new Funeral Parlour prices (he hasn't bought them up yet, but give him time).

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PostPosted: Thu Jul 09, 2020 12:53 am    Post subject: Reply with quote

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"The more accepting people are at the beginning of totalitarianism, the more horribly they will suffer at the end. Yes, it is accepted because their fear is being manipulated: they fear the named danger - either terrorists or viruses - more than they fear the smooth liars who say they are the saviors. But in the end, when the masks finally come off, they will realize that what they were manipulated to fear was nothing, and it was the wolf in sheep's clothing that is now going to devour them and their children.

The Germans could have resisted Hitler at the beginning, but they didn't, and Germany was almost entirely destroyed as a consequence. Today, it is the Liberal Left globalists who are the new Nazis." ..... A Critical Thinker

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PostPosted: Mon Jul 13, 2020 9:21 am    Post subject: Reply with quote

Whitehall_Bin_Men wrote:
Excess winter mortality in England and Wales: 2017 to 2018 (provisional) and 2016 to 2017 (final)
https://www.ons.gov.uk/peoplepopulationandcommunity/birthsdeathsandmar riages/deaths/bulletins/excesswintermortalityinenglandandwales/2017to2 018provisionaland2016to2017final

More people die in the winter than the summer. We present data by sex, age, region and cause of death.

This is not the latest release. View latest release

This is an accredited national statistic.Contact:
Melissa Bennett Release date:
30 November 2018
Next release:
November 2019
Table of contents
Main points
Statistician’s comment
Things you need to know about this release
Excess winter deaths in 2017 to 2018 are the highest recorded since the winter of 1975 to 1976
Pronounced increase in mortality over the winter months
Females and the elderly were most affected by excess winter mortality
Respiratory disease caused most excess winter deaths
Predominant strain of influenza in winter 2017 to 2018 impacted the elderly the most
Relationship between excess winter mortality and temperature is complex
Excess winter mortality significantly increased in all regions of England and in Wales in 2017 to 2018
What causes excess winter mortality?
Links to related statistics
Quality and methodology
Print this statistical bulletin

Download as PDF

View all data used in this statistical bulletin
1. Main points

In the 2017 to 2018 winter period, there were an estimated 50,100 excess winter deaths in England and Wales.
The number of excess winter deaths in 2017 to 2018 was the highest recorded since winter 1975 to 1976.
During the winter months of 2017 to 2018, the number of daily deaths exceeded the daily five-year average for all days except 25 March.
Excess winter mortality in 2017 to 2018 significantly increased from 2016 to 2017 in all English regions and Wales, with Wales having the highest regional index.
Excess winter mortality continued to be highest in females and people aged 85 and over.
Excess winter mortality doubled among males aged 0 to 64 years between 2016 to 2017 and 2017 to 2018.
Over one-third (34.7%) of all excess winter deaths were caused by respiratory diseases.
Back to table of contents
2. Statistician’s comment

“The number of excess winter deaths in England and Wales in 2017 to 2018 was the highest recorded since the winter of 1975 to 1976. However, peaks like these are not unusual – we have seen more than eight peaks during the last 40 years. It is likely that last winter’s increase was due to the predominant strain of flu, the effectiveness of the influenza vaccine and below-average winter temperatures”.

Nick Stripe, Health Analysis and Life Events, Office for National Statistics

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Martin Van Creveld: Let me quote General Moshe Dayan: "Israel must be like a mad dog, too dangerous to bother."
Martin Van Creveld: I'll quote Henry Kissinger: "In campaigns like this the antiterror forces lose, because they don't win, and the rebels win by not losing."
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PostPosted: Mon Jul 13, 2020 9:45 am    Post subject: Reply with quote

Ep.132 | The Parasitic Occult Initiation Rituals

Link

https://www.youtube.com/watch?v=uoRXvCnZ484

Adrian 15.8K subscribers Published on 11 Jul 2020

Awareness can break the spell. We are witnessing a massive social engineering project to completely change everyone. Well, that appears to be the plan. It's worth looking into this so you don't fall victim and become a sleep walker like so many.

Article I was referencing
Occult Ritual Transformation and Coronavirus: How Mask Wearing, Hand Washing, “Social Separation” and Lockdowns Are Age-Old Occult Rituals Being Used to Initiate People Into a New Global Order
https://haveyenotread.com/occult-ritu...

The characteristics of an initiation ritual
https://www.youtube.com/watch?time_co...

The Isolated individual and the crowd beast
https://www.youtube.com/watch?v=bBgbI...

Alice in Wonderland
https://www.bitchute.com/video/mYKIp5...


As always please do your own research, thinking and draw your own conclusions.

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_________________
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'Suppression of truth, human spirit and the holy chord of justice never works long-term. Something the suppressors never get.' David Southwell
http://aangirfan.blogspot.com
http://aanirfan.blogspot.com
Martin Van Creveld: Let me quote General Moshe Dayan: "Israel must be like a mad dog, too dangerous to bother."
Martin Van Creveld: I'll quote Henry Kissinger: "In campaigns like this the antiterror forces lose, because they don't win, and the rebels win by not losing."
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PostPosted: Tue Jul 14, 2020 11:10 pm    Post subject: Reply with quote

Horowitz: The CDC Confirms Remarkably Low Coronavirus Death Rate. Where is the Media?
https://www.greenmedinfo.com/blog/horowitz-cdc-confirms-remarkably-low -coronavirus-death-rate-where-media

Posted on: Wednesday, July 8th 2020 at 1:15 pm
Written By: Daniel Horowitz

Originally published on www.conservativereview.com

Most people are more likely to wind up six feet under because of almost anything else under the sun other than COVID-19.

The CDC just came out with a report that should be earth-shattering to the narrative of the political class, yet it will go into the thick pile of vital data and information about the virus that is not getting out to the public. For the first time, the CDC has attempted to offer a real estimate of the overall death rate for COVID-19, and under its most likely scenario, the number is 0.26%. Officials estimate a 0.4% fatality rate among those who are symptomatic and project a 35% rate of asymptomatic cases among those infected, which drops the overall infection fatality rate (IFR) to just 0.26% -- almost exactly where Stanford researchers pegged it a month ago.





Until now, we have been ridiculed for thinking the death rate was that low, as opposed to the 3.4% estimate of the World Health Organization, which helped drive the panic and the lockdowns. Now the CDC is agreeing to the lower rate in plain ink.

Plus, ultimately we might find out that the IFR is even lower because numerous studies and hard counts of confined populations have shown a much higher percentage of asymptomatic cases. Simply adjusting for a 50% asymptomatic rate would drop their fatality rate to 0.2% - exactly the rate of fatality Dr. John Ionnidis of Stanford University projected.

More importantly, as I mentioned before, the overall death rate is meaningless because the numbers are so lopsided. Given that at least half of the deaths were in nursing homes, a back-of-the-envelope estimate would show that the infection fatality rate for non-nursing home residents would only be 0.1% or 1 in 1,000. And that includes people of all ages and all health statuses outside of nursing homes. Since nearly all of the deaths are those with comorbidities.

The CDC estimates the death rate from COVID-19 for those under 50 is 1 in 5,000 for those with symptoms, which would be 1 in 6,725 overall, but again, almost all those who die have specific comorbidities or underlying conditions. Those without them are more likely to die in a car accident. And schoolchildren, whose lives, mental health, and education we are destroying, are more likely to get struck by lightning.

To put this in perspective, one Twitter commentator juxtaposed the age-separated infection fatality rates in Spain to the average yearly probability of dying of anything for the same age groups, based on data from the Social Security Administration. He used Spain because we don't have a detailed infection fatality rate estimate for each age group from any survey in the U.S. However, we know that Spain fared worse than almost every other country. This data is actually working with a top-line IFR of 1%, roughly four times what the CDC estimates for the U.S., so if anything, the corresponding numbers for the U.S. will be lower.



As you can see, even in Spain, the death rates from COVID-19 for younger people are very low and are well below the annual death rate for any age group in a given year. For children, despite their young age, they are 10-30 times more likely to die from other causes in any given year.

While obviously yearly death rates factor in myriad of causes of death and COVID-19 is just one virus, it still provides much-needed perspective to a public policy response that is completely divorced from the risk for all but the oldest and sickest people in the country.

Also, keep in mind, these numbers represent your chance of dying once you have already contracted the virus, aka the infection fatality rate. Once you couple the chance of contracting the virus in the first place together with the chance of dying from it, many younger people have a higher chance of dying from a lightning strike.

Four infectious disease doctors in Canada estimate that the individual rate of death from COVID-19 for people under 65 years of age is six per million people, or 0.0006 per cent - 1 in 166,666, which is "roughly equivalent to the risk of dying from a motor vehicle accident during the same time period." These numbers are for Canada, which did have fewer deaths per capita than the U.S.; however, if you take New York City and its surrounding counties out of the equation, the two countries are pretty much the same. Also, remember, so much of the death is associated with the suicidal political decisions of certain states and countries to place COVID-19 patients in nursing homes. An astounding 62 percent of all COVID-19 deaths were in the six states confirmed to have done this, even though they only compose 18 percent of the national population.

We destroyed our entire country and suspended democracy all for a lie, and these people perpetrated the unscientific degree of panic. Will they ever admit the grave consequences of their error?

Daniel Horowitz is a senior editor of Conservative Review. Follow him on Twitter @RMConservative.

_________________
--
'Suppression of truth, human spirit and the holy chord of justice never works long-term. Something the suppressors never get.' David Southwell
http://aangirfan.blogspot.com
http://aanirfan.blogspot.com
Martin Van Creveld: Let me quote General Moshe Dayan: "Israel must be like a mad dog, too dangerous to bother."
Martin Van Creveld: I'll quote Henry Kissinger: "In campaigns like this the antiterror forces lose, because they don't win, and the rebels win by not losing."
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PostPosted: Fri Jul 17, 2020 1:15 am    Post subject: Reply with quote

Coronavirus is ARK: http://www.911forum.org.uk/board/viewtopic.php?t=24304
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PostPosted: Sat Jul 18, 2020 9:38 pm    Post subject: Reply with quote


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PostPosted: Wed Jul 22, 2020 11:50 pm    Post subject: Reply with quote

Lockdown may cost 200,000 lives, government report shows
Research shines a light on the reasons why the Government has been keen to lift lockdown, in spite of experts claiming it happened too soon
https://www.telegraph.co.uk/news/2020/07/19/lockdown-may-cost-200k-liv es-government-report-shows/

By Sarah Knapton, SCIENCE EDITOR 19 July 2020 • 9:00pm

More than 200,000 people could die from the impact of lockdown and protecting the NHS, an official government report shows.

As national restrictions were imposed, experts from the Department of Health, the Office of National Statistics (ONS), the government’s Actuary Department and the Home Office forecast the collateral damage from delays to healthcare and the effects of recession arising from the pandemic response.

It estimated that in a reasonable worst case scenario, around 50,000 people would die from coronavirus in the first six months of the pandemic, with mitigation measures in place.

But in the report published in April they calculated that up to 25,000 could die from delays to treatment in the same period and a further 185,000 in the medium to long term - amounting to nearly one million years of life lost.

It comes amid debate over the easing of lockdown restrictions, with some arguing it is both too early to lift the measures and that they should have been imposed earlier, while other politicians have questioned whether the cure is worse than the disease.


The Prime Minister has stressed his desire to avoid a return to national lockdown.

In an interview in The Telegraph, Boris Johnson likened the measures to a nuclear deterrent, and said he did not want to impose blanket restrictions again, or think it would be necessary.

The UK's National Statistician, Prof Sir Ian Diamond also said on Sunday that there had been no uptick in cases since lockdown measures had been eased but warned the nation would need to be vigilant come the autumn.

Professor Chris Whitty, the Chief Medical Officer (CMO), and Sir Patrick Vallance, the government’s chief scientific advisor (GSCA) have both expressed concern that the damage from lockdown could be severe.

The report came to light after Sir Patrick told MPs at the Science and Technology Select Committee last week that calculations had been made to predict the number of deaths caused by the effects of lockdown, which was announced on March 23.

The report produced in the following weeks warned there could be 500 more suicides during the first wave, and between 600 and 12,000 more deaths per year resulting from a recession which had a significant impact on GDP.

They also forecast around 20 more deaths this year through domestic violence, and an increase in the number of accidents at home - in the ‘low tens.’ In total, under a worst case scenario, around quarter of a million people would die because of the pandemic response.

The figures were based on 75 per cent of elective care being cancelled over six months without significant reprioritisation when things returned to normal.

The number of elective hospital appointments dropped to around a quarter of usual levels in March and April and had only recovered to around half by the beginning of July.

Charities have increasingly warned that delaying diagnosis, pausing surgery and postponing treatment is a ‘ticking time bomb’ which will cause long-term harm.


Figures released by Cancer Research UK today show that as of May 30, there were more than 180,000 people in England waiting for an endoscopy - a rise of 44 per cent from the same time in 2019.

And around 2.3 million fewer tests that help diagnose cancer have taken place since lockdown compared to the same time last year, and 51 per cent more people are waiting for colonoscopies.

Michelle Mitchell, Cancer Research UK’s chief executive, said: “Covid-19 has had a devastating impact on cancer patients and services across the UK.

“In the early weeks of lockdown urgent referrals plummeted, screening programmes were paused, surgeries were cancelled, clinical trials were put on hold, and existing health inequalities were exacerbated.

“It’s now more crucial than ever that the Government works closely with the NHS to ensure it has the staff and equipment it needs to clear the mounting backlog and get services back on track before this situation gets even worse – especially in the event of a second wave.”

The report points out that nearly 500,000 people would have died from coronavirus if the virus had been allowed to run through the population unchecked. And there would have been more than a million non-Covid deaths resulting from missed treatment if the health service had been overwhelmed in dealing with the pandemic.


But charities said more should have been done to get medical care up and running quickly when it became clear early on that the NHS was not stretched. Nightingale Hospitals in particular were largely empty even at the peak of the pandemic and have only recently been reassigned for normal care.

Dr Sonya Babu-Narayan, Associate Medical Director at the British Heart Foundation and Consultant Cardiologist, said it was a priority to restore heart and circulatory care: “This report is a sobering wake-up call for governments and the NHS across the UK to urgently restore and maintain care for people with medical conditions including heart and circulatory diseases.

“Although the latest statistics have shown fewer excess deaths in recent weeks, people are still experiencing delays in accessing vital treatment and care, which could make them sicker and ultimately lead to more deaths from both undiagnosed and existing conditions.

“The growing and significant backlog must be tackled to prevent a tidal wave of illness overwhelming the NHS whilst it’s fighting Covid-19.”

Last week, the University of Oxford found that 5,000 fewer heart attack patients had attended hospital from March to May, many of whom could have died through lack of medical care.

The Institute of Cancer Research (ICR) also warned that delays in diagnosis and treatment meant many people who would have recovered from their illness were now facing incurable cancer.

Modelling by the ICR suggests that a three-month delay to surgery alone across all Stage 1-3 cancers could cause more than 4,700 attributable deaths per year in England.

Clare Turnbull, Professor of Cancer Genomics at The Institute of Cancer Research, London, said: “Lockdown and re-deployment of NHS workers as a result of the COVID-19 pandemic is causing significant disruption to cancer diagnosis and management.

“For patients with cancer, delay of surgery has the real potential to increase the likelihood of advanced disease, with some patients’ tumours progressing from being curable – with near normal life expectancy – to non-curable – with limited life expectancy.

“At this point, it is crucial to ramp up capacity as quickly as possible to allow cancer services to clear the backlog accumulated during lockdown.”

Figures released last week from Target Ovarian Cancer showed that more than half of women with ovarian cancer surveyed said their treatment had been affected by coronavirus.

Annwen Jones OBE, Chief Executive of Target Ovarian Cancer, said: “It’s hard to overstate the difficulties faced by women with ovarian cancer during this pandemic. We must not let them down as health services recover.

“It is urgent that we now see comprehensive plans and a timeline for the full restoration of diagnostic, treatment and support services. This is the only way to avoid worse outcomes and a devastating toll on women’s lives.”

Data compiled by The Telegraph showed that by the end of May there had been more than 23,000 excess deaths in care homes or at home, not linked to Covid-19, since the middle of March.

There are also fears that the government’s ‘protect the NHS’ message was too effective, with many people staying away from hospitals when they needed urgent medical help.

In April, Chief Executive of NHS England Sir Simon Stevens was forced to issue an urgent message asking people to come forward if they needed treatment.

However the report did predict some good news for lockdown. Experts calculated that there would be approximately 200 - 500 fewer road traffic and air pollution deaths, 67 fewer murders and a small drop in work related accidents during the six month first wave period.

There may also be a positive impact on health as a result of increased physical activity as people take the opportunity to use their one instance of daily exercise, the authors state, although they also warn that some may become more sedentary whilst staying home. And there is some evidence that recessions can improve mortality rates, possibly saving a few thousands lives.

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PostPosted: Fri Jul 24, 2020 12:53 pm    Post subject: Reply with quote

Mask wearing experience of asthma patient today


Link


https://youtu.be/ki6ztpQ_M18

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Martin Van Creveld: Let me quote General Moshe Dayan: "Israel must be like a mad dog, too dangerous to bother."
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PostPosted: Tue Jul 28, 2020 11:46 am    Post subject: Reply with quote


Link


http://www.youtube.com/watch?v=xfyLceO_Vf4



COVID-19
Get the latest information from the NHS about coronavirus.

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See more resources on Google


DOCTORS SPEAK OUT ABOUT COVID & THE #FearPorn #FakeNews
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Published on 27 Jul 2020
#Plandemic #Hydroxychloroquine #CovidCure

BREAKING: American Doctors Address COVID-19 Misinformation with SCOTUS Press Conference...
FINALLY...REAL FRONTLINE DOCTORS TELLING THE TRUTH

_________________
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'Suppression of truth, human spirit and the holy chord of justice never works long-term. Something the suppressors never get.' David Southwell
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http://aanirfan.blogspot.com
Martin Van Creveld: Let me quote General Moshe Dayan: "Israel must be like a mad dog, too dangerous to bother."
Martin Van Creveld: I'll quote Henry Kissinger: "In campaigns like this the antiterror forces lose, because they don't win, and the rebels win by not losing."
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PostPosted: Sun Aug 02, 2020 9:48 am    Post subject: Reply with quote

Chloroquine is a potent inhibitor of SARS coronavirus infection and spread
https://virologyj.biomedcentral.com/articles/10.1186/1743-422X-2-69

Martin J Vincent, Eric Bergeron, […]Stuart T Nichol
Virology Journal volume 2, Article number: 69 (2005) Cite this article

401k Accesses
378 Citations
40607 Altmetric
Metrics details
Abstract

Background
Severe acute respiratory syndrome (SARS) is caused by a newly discovered coronavirus (SARS-CoV). No effective prophylactic or post-exposure therapy is currently available.

Results
We report, however, that chloroquine has strong antiviral effects on SARS-CoV infection of primate cells. These inhibitory effects are observed when the cells are treated with the drug either before or after exposure to the virus, suggesting both prophylactic and therapeutic advantage. In addition to the well-known functions of chloroquine such as elevations of endosomal pH, the drug appears to interfere with terminal glycosylation of the cellular receptor, angiotensin-converting enzyme 2. This may negatively influence the virus-receptor binding and abrogate the infection, with further ramifications by the elevation of vesicular pH, resulting in the inhibition of infection and spread of SARS CoV at clinically admissible concentrations.

Conclusion
Chloroquine is effective in preventing the spread of SARS CoV in cell culture. Favorable inhibition of virus spread was observed when the cells were either treated with chloroquine prior to or after SARS CoV infection. In addition, the indirect immunofluorescence assay described herein represents a simple and rapid method for screening SARS-CoV antiviral compounds.

Background

Severe acute respiratory syndrome (SARS) is an emerging disease that was first reported in Guangdong Province, China, in late 2002. The disease rapidly spread to at least 30 countries within months of its first appearance, and concerted worldwide efforts led to the identification of the etiological agent as SARS coronavirus (SARS-CoV), a novel member of the family Coronaviridae [1]. Complete genome sequencing of SARS-CoV [2, 3] confirmed that this pathogen is not closely related to any of the previously established coronavirus groups. Budding of the SARS-CoV occurs in the Golgi apparatus [4] and results in the incorporation of the envelope spike glycoprotein into the virion. The spike glycoprotein is a type I membrane protein that facilitates viral attachment to the cellular receptor and initiation of infection, and angiotensin-converting enzyme-2 (ACE2) has been identified as a functional cellular receptor of SARS-CoV [5]. We have recently shown that the processing of the spike protein was effected by furin-like convertases and that inhibition of this cleavage by a specific inhibitor abrogated cytopathicity and significantly reduced the virus titer of SARS-CoV [6].

Due to the severity of SARS-CoV infection, the potential for rapid spread of the disease, and the absence of proven effective and safe in vivo inhibitors of the virus, it is important to identify drugs that can effectively be used to treat or prevent potential SARS-CoV infections. Many novel therapeutic approaches have been evaluated in laboratory studies of SARS-CoV: notable among these approaches are those using siRNA [7], passive antibody transfer [8], DNA vaccination [9], vaccinia or parainfluenza virus expressing the spike protein [10, 11], interferons [12, 13], and monoclonal antibody to the S1-subunit of the spike glycoprotein that blocks receptor binding [14]. In this report, we describe the identification of chloroquine as an effective pre- and post-infection antiviral agent for SARS-CoV. Chloroquine, a 9-aminoquinoline that was identified in 1934, is a weak base that increases the pH of acidic vesicles. When added extracellularly, the non-protonated portion of chloroquine enters the cell, where it becomes protonated and concentrated in acidic, low-pH organelles, such as endosomes, Golgi vesicles, and lysosomes. Chloroquine can affect virus infection in many ways, and the antiviral effect depends in part on the extent to which the virus utilizes endosomes for entry. Chloroquine has been widely used to treat human diseases, such as malaria, amoebiosis, HIV, and autoimmune diseases, without significant detrimental side effects [15]. Together with data presented here, showing virus inhibition in cell culture by chloroquine doses compatible with patient treatment, these features suggest that further evaluation of chloroquine in animal models of SARS-CoV infection would be warranted as we progress toward finding effective antivirals for prevention or treatment of the disease.

Results

Preinfection chloroquine treatment renders Vero E6 cells refractory to SARS-CoV infection
In order to investigate if chloroquine might prevent SARS-CoV infection, permissive Vero E6 cells [1] were pretreated with various concentrations of chloroquine (0.1–10 μM) for 20–24 h prior to virus infection. Cells were then infected with SARS-CoV, and virus antigens were visualized by indirect immunofluorescence as described in Materials and Methods. Microscopic examination (Fig. 1A) of the control cells (untreated, infected) revealed extensive SARS-CoV-specific immunostaining of the monolayer. A dose-dependant decrease in virus antigen-positive cells was observed starting at 0.1 μM chloroquine, and concentrations of 10 μM completely abolished SARS-CoV infection. For quantitative purposes, we counted the number of cells stained positive from three random locations on a slide. The average number of positively stained control cells was scored as 100% and was compared with the number of positive cells observed under various chloroquine concentrations (Fig. 1B). Pretreatment with 0.1, 1, and 10 μM chloroquine reduced infectivity by 28%, 53%, and 100%, respectively. Reproducible results were obtained from three independent experiments. These data demonstrated that pretreatment of Vero E6 cells with chloroquine rendered these cells refractory to SARS-CoV infection.

_________________
--
'Suppression of truth, human spirit and the holy chord of justice never works long-term. Something the suppressors never get.' David Southwell
http://aangirfan.blogspot.com
http://aanirfan.blogspot.com
Martin Van Creveld: Let me quote General Moshe Dayan: "Israel must be like a mad dog, too dangerous to bother."
Martin Van Creveld: I'll quote Henry Kissinger: "In campaigns like this the antiterror forces lose, because they don't win, and the rebels win by not losing."
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PostPosted: Sun Aug 02, 2020 9:48 am    Post subject: Reply with quote

Chloroquine is a potent inhibitor of SARS coronavirus infection and spread
https://virologyj.biomedcentral.com/articles/10.1186/1743-422X-2-69

Martin J Vincent, Eric Bergeron, […]Stuart T Nichol
Virology Journal volume 2, Article number: 69 (2005) Cite this article

401k Accesses
378 Citations
40607 Altmetric
Metrics details
Abstract

Background
Severe acute respiratory syndrome (SARS) is caused by a newly discovered coronavirus (SARS-CoV). No effective prophylactic or post-exposure therapy is currently available.

Results
We report, however, that chloroquine has strong antiviral effects on SARS-CoV infection of primate cells. These inhibitory effects are observed when the cells are treated with the drug either before or after exposure to the virus, suggesting both prophylactic and therapeutic advantage. In addition to the well-known functions of chloroquine such as elevations of endosomal pH, the drug appears to interfere with terminal glycosylation of the cellular receptor, angiotensin-converting enzyme 2. This may negatively influence the virus-receptor binding and abrogate the infection, with further ramifications by the elevation of vesicular pH, resulting in the inhibition of infection and spread of SARS CoV at clinically admissible concentrations.

Conclusion
Chloroquine is effective in preventing the spread of SARS CoV in cell culture. Favorable inhibition of virus spread was observed when the cells were either treated with chloroquine prior to or after SARS CoV infection. In addition, the indirect immunofluorescence assay described herein represents a simple and rapid method for screening SARS-CoV antiviral compounds.

Background

Severe acute respiratory syndrome (SARS) is an emerging disease that was first reported in Guangdong Province, China, in late 2002. The disease rapidly spread to at least 30 countries within months of its first appearance, and concerted worldwide efforts led to the identification of the etiological agent as SARS coronavirus (SARS-CoV), a novel member of the family Coronaviridae [1]. Complete genome sequencing of SARS-CoV [2, 3] confirmed that this pathogen is not closely related to any of the previously established coronavirus groups. Budding of the SARS-CoV occurs in the Golgi apparatus [4] and results in the incorporation of the envelope spike glycoprotein into the virion. The spike glycoprotein is a type I membrane protein that facilitates viral attachment to the cellular receptor and initiation of infection, and angiotensin-converting enzyme-2 (ACE2) has been identified as a functional cellular receptor of SARS-CoV [5]. We have recently shown that the processing of the spike protein was effected by furin-like convertases and that inhibition of this cleavage by a specific inhibitor abrogated cytopathicity and significantly reduced the virus titer of SARS-CoV [6].

Due to the severity of SARS-CoV infection, the potential for rapid spread of the disease, and the absence of proven effective and safe in vivo inhibitors of the virus, it is important to identify drugs that can effectively be used to treat or prevent potential SARS-CoV infections. Many novel therapeutic approaches have been evaluated in laboratory studies of SARS-CoV: notable among these approaches are those using siRNA [7], passive antibody transfer [8], DNA vaccination [9], vaccinia or parainfluenza virus expressing the spike protein [10, 11], interferons [12, 13], and monoclonal antibody to the S1-subunit of the spike glycoprotein that blocks receptor binding [14]. In this report, we describe the identification of chloroquine as an effective pre- and post-infection antiviral agent for SARS-CoV. Chloroquine, a 9-aminoquinoline that was identified in 1934, is a weak base that increases the pH of acidic vesicles. When added extracellularly, the non-protonated portion of chloroquine enters the cell, where it becomes protonated and concentrated in acidic, low-pH organelles, such as endosomes, Golgi vesicles, and lysosomes. Chloroquine can affect virus infection in many ways, and the antiviral effect depends in part on the extent to which the virus utilizes endosomes for entry. Chloroquine has been widely used to treat human diseases, such as malaria, amoebiosis, HIV, and autoimmune diseases, without significant detrimental side effects [15]. Together with data presented here, showing virus inhibition in cell culture by chloroquine doses compatible with patient treatment, these features suggest that further evaluation of chloroquine in animal models of SARS-CoV infection would be warranted as we progress toward finding effective antivirals for prevention or treatment of the disease.

Results

Preinfection chloroquine treatment renders Vero E6 cells refractory to SARS-CoV infection
In order to investigate if chloroquine might prevent SARS-CoV infection, permissive Vero E6 cells [1] were pretreated with various concentrations of chloroquine (0.1–10 μM) for 20–24 h prior to virus infection. Cells were then infected with SARS-CoV, and virus antigens were visualized by indirect immunofluorescence as described in Materials and Methods. Microscopic examination (Fig. 1A) of the control cells (untreated, infected) revealed extensive SARS-CoV-specific immunostaining of the monolayer. A dose-dependant decrease in virus antigen-positive cells was observed starting at 0.1 μM chloroquine, and concentrations of 10 μM completely abolished SARS-CoV infection. For quantitative purposes, we counted the number of cells stained positive from three random locations on a slide. The average number of positively stained control cells was scored as 100% and was compared with the number of positive cells observed under various chloroquine concentrations (Fig. 1B). Pretreatment with 0.1, 1, and 10 μM chloroquine reduced infectivity by 28%, 53%, and 100%, respectively. Reproducible results were obtained from three independent experiments. These data demonstrated that pretreatment of Vero E6 cells with chloroquine rendered these cells refractory to SARS-CoV infection.



https://seed306.bitchute.com/FqVwIm1CMAby/m6Ka8wVS5CeJ.mp4



https://seed167.bitchute.com/FqVwIm1CMAby/Dd3L0Jr85Nfi.mp4

_________________
--
'Suppression of truth, human spirit and the holy chord of justice never works long-term. Something the suppressors never get.' David Southwell
http://aangirfan.blogspot.com
http://aanirfan.blogspot.com
Martin Van Creveld: Let me quote General Moshe Dayan: "Israel must be like a mad dog, too dangerous to bother."
Martin Van Creveld: I'll quote Henry Kissinger: "In campaigns like this the antiterror forces lose, because they don't win, and the rebels win by not losing."
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PostPosted: Sun Aug 02, 2020 1:32 pm    Post subject: Reply with quote

100%. His arguments and points are bullet proof. An honest comment from and honest journalist.

Link

https://www.youtube.com/watch?v=ZUU7Q92aPgY


Heated Vaccine Debate - Kennedy Jr. vs Dershowitz

Link

https://www.youtube.com/watch?v=IfnJi7yLKgE

_________________
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www.v911t.org
www.thisweek.org.uk
www.abolishwar.org.uk
www.elementary.org.uk
www.radio4all.net/index.php/contributor/2149
http://utangente.free.fr/2003/media2003.pdf
"The maintenance of secrets acts like a psychic poison which alienates the possessor from the community" Carl Jung
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PostPosted: Mon Aug 03, 2020 12:28 am    Post subject: Reply with quote

Flu vaccine associated with a 26.5% increase in pneumonia-influenza mortality

Overall, the lifesaving value of seasonal influenza vaccines is uncertain.
https://www.bmj.com/content/369/bmj.m1932/rr-24

This fact was recently highlighted by a study that looked at mortality in the elderly over a 14-year period. (Anderson et al, Ann Intern Med 2020;172:445)
Table 2 indicates that influenza vaccine was associated with an 8.9% increase in all-cause mortality in elderly men (VE­8.9%, CI –19.6% to 1.8%), and a 26.5% increase in pneumonia-influenza mortality (VE –26.5%, CI –56.1% to 3.0%)

ALLAN S. CUNNINGHAM 28 July 2020

Competing interests: No competing interests

_________________
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www.mp911truth.org
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www.rl911truth.org
www.stj911.org
www.v911t.org
www.thisweek.org.uk
www.abolishwar.org.uk
www.elementary.org.uk
www.radio4all.net/index.php/contributor/2149
http://utangente.free.fr/2003/media2003.pdf
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PostPosted: Mon Aug 03, 2020 11:05 pm    Post subject: Reply with quote

Chloroquine is a potent inhibitor of SARS coronavirus infection and spread
Martin J Vincent, Eric Bergeron, Suzanne Benjannet, Bobbie R Erickson, Pierre E Rollin, Thomas G Ksiazek, Nabil G Seidah & Stuart T Nichol
Virology Journal volume 2, Article number: 69 (2005) Cite this article

401k Accesses
https://virologyj.biomedcentral.com/articles/10.1186/1743-422X-2-69

Abstract
Background
Severe acute respiratory syndrome (SARS) is caused by a newly discovered coronavirus (SARS-CoV). No effective prophylactic or post-exposure therapy is currently available.

Results
We report, however, that chloroquine has strong antiviral effects on SARS-CoV infection of primate cells. These inhibitory effects are observed when the cells are treated with the drug either before or after exposure to the virus, suggesting both prophylactic and therapeutic advantage. In addition to the well-known functions of chloroquine such as elevations of endosomal pH, the drug appears to interfere with terminal glycosylation of the cellular receptor, angiotensin-converting enzyme 2. This may negatively influence the virus-receptor binding and abrogate the infection, with further ramifications by the elevation of vesicular pH, resulting in the inhibition of infection and spread of SARS CoV at clinically admissible concentrations.

Conclusion
Chloroquine is effective in preventing the spread of SARS CoV in cell culture. Favorable inhibition of virus spread was observed when the cells were either treated with chloroquine prior to or after SARS CoV infection. In addition, the indirect immunofluorescence assay described herein represents a simple and rapid method for screening SARS-CoV antiviral compounds.

Background
Severe acute respiratory syndrome (SARS) is an emerging disease that was first reported in Guangdong Province, China, in late 2002. The disease rapidly spread to at least 30 countries within months of its first appearance, and concerted worldwide efforts led to the identification of the etiological agent as SARS coronavirus (SARS-CoV), a novel member of the family Coronaviridae [1]. Complete genome sequencing of SARS-CoV [2, 3] confirmed that this pathogen is not closely related to any of the previously established coronavirus groups. Budding of the SARS-CoV occurs in the Golgi apparatus [4] and results in the incorporation of the envelope spike glycoprotein into the virion. The spike glycoprotein is a type I membrane protein that facilitates viral attachment to the cellular receptor and initiation of infection, and angiotensin-converting enzyme-2 (ACE2) has been identified as a functional cellular receptor of SARS-CoV [5]. We have recently shown that the processing of the spike protein was effected by furin-like convertases and that inhibition of this cleavage by a specific inhibitor abrogated cytopathicity and significantly reduced the virus titer of SARS-CoV [6].

Due to the severity of SARS-CoV infection, the potential for rapid spread of the disease, and the absence of proven effective and safe in vivo inhibitors of the virus, it is important to identify drugs that can effectively be used to treat or prevent potential SARS-CoV infections. Many novel therapeutic approaches have been evaluated in laboratory studies of SARS-CoV: notable among these approaches are those using siRNA [7], passive antibody transfer [8], DNA vaccination [9], vaccinia or parainfluenza virus expressing the spike protein [10, 11], interferons [12, 13], and monoclonal antibody to the S1-subunit of the spike glycoprotein that blocks receptor binding [14]. In this report, we describe the identification of chloroquine as an effective pre- and post-infection antiviral agent for SARS-CoV. Chloroquine, a 9-aminoquinoline that was identified in 1934, is a weak base that increases the pH of acidic vesicles. When added extracellularly, the non-protonated portion of chloroquine enters the cell, where it becomes protonated and concentrated in acidic, low-pH organelles, such as endosomes, Golgi vesicles, and lysosomes. Chloroquine can affect virus infection in many ways, and the antiviral effect depends in part on the extent to which the virus utilizes endosomes for entry. Chloroquine has been widely used to treat human diseases, such as malaria, amoebiosis, HIV, and autoimmune diseases, without significant detrimental side effects [15]. Together with data presented here, showing virus inhibition in cell culture by chloroquine doses compatible with patient treatment, these features suggest that further evaluation of chloroquine in animal models of SARS-CoV infection would be warranted as we progress toward finding effective antivirals for prevention or treatment of the disease.

Results
Preinfection chloroquine treatment renders Vero E6 cells refractory to SARS-CoV infection
In order to investigate if chloroquine might prevent SARS-CoV infection, permissive Vero E6 cells [1] were pretreated with various concentrations of chloroquine (0.1–10 μM) for 20–24 h prior to virus infection. Cells were then infected with SARS-CoV, and virus antigens were visualized by indirect immunofluorescence as described in Materials and Methods. Microscopic examination (Fig. 1A) of the control cells (untreated, infected) revealed extensive SARS-CoV-specific immunostaining of the monolayer. A dose-dependant decrease in virus antigen-positive cells was observed starting at 0.1 μM chloroquine, and concentrations of 10 μM completely abolished SARS-CoV infection. For quantitative purposes, we counted the number of cells stained positive from three random locations on a slide. The average number of positively stained control cells was scored as 100% and was compared with the number of positive cells observed under various chloroquine concentrations (Fig. 1B). Pretreatment with 0.1, 1, and 10 μM chloroquine reduced infectivity by 28%, 53%, and 100%, respectively. Reproducible results were obtained from three independent experiments. These data demonstrated that pretreatment of Vero E6 cells with chloroquine rendered these cells refractory to SARS-CoV infection.

Figure 1
figure1
Prophylactic effect of chloroquine. Vero E6 cells pre-treated with chloroquine for 20 hrs. Chloroquine-containing media were removed and the cells were washed with phosphate buffered saline before they were infected with SARS-CoV (0.5 multiplicity of infection) for 1 h. in the absence of chloroquine. Virus was then removed and the cells were maintained in Opti-MEM (Invitrogen) for 16–18 h in the absence of chloroquine. SARS-CoV antigens were stained with virus-specific HMAF, followed by FITC-conjugated secondary antibodies. (A) The concentration of chloroquine used is indicated on the top of each panel. (B) SARS-CoV antigen-positive cells at three random locations were captured by using a digital camera, the number of antigen-positive cells was determined, and the average inhibition was calculated. Percent inhibition was obtained by considering the untreated control as 0% inhibition. The vertical bars represent the range of SEM.

Full size image
Postinfection chloroquine treatment is effective in preventing the spread of SARS-CoV infection
In order to investigate the antiviral properties of chloroquine on SARS-CoV after the initiation of infection, Vero E6 cells were infected with the virus and fresh medium supplemented with various concentrations of chloroquine was added immediately after virus adsorption. Infected cells were incubated for an additional 16–18 h, after which the presence of virus antigens was analyzed by indirect immunofluorescence analysis. When chloroquine was added after the initiation of infection, there was a dramatic dose-dependant decrease in the number of virus antigen-positive cells (Fig. 2A). As little as 0.1–1 μM chloroquine reduced the infection by 50% and up to 90–94% inhibition was observed with 33–100 μM concentrations (Fig. 2B). At concentrations of chloroquine in excess of 1 μM, only a small number of individual cells were initially infected, and the spread of the infection to adjacent cells was all but eliminated. A half-maximal inhibitory effect was estimated to occur at 4.4 ± 1.0 μM chloroquine (Fig. 2C). These data clearly show that addition of chloroquine can effectively reduce the establishment of infection and spread of SARS-CoV if the drug is added immediately following virus adsorption.

Figure 2
figure2
Post-infection chloroquine treatment reduces SARS-CoV infection and spread. Vero E6 cells were seeded and infected as described for Fig. 1 except that chloroquine was added only after virus adsorption. Cells were maintained in Opti-MEM (Invitrogen) containing chloroquine for 16–18 h, after which they were processed for immunofluorescence. (A) The concentration of chloroquine is indicated on the top. (B) Percent inhibition and SEM were calculated as in Fig. 1B. (C) The effective dose (ED50) was calculated using commercially available software (Grafit, version 4, Erithacus Software).

Full size image
Electron microscopic analysis indicated the appearance of significant amounts of extracellular virus particles 5–6 h after infection [16]. Since we observed antiviral effects by chloroquine immediately after virus adsorption, we further extended the analysis by adding chloroquine 3 and 5 h after virus adsorption and examined for the presence of virus antigens after 20 h. We found that chloroquine was still significantly effective even when added 5 h after infection (Fig. 3); however, to obtain equivalent antiviral effect, a higher concentration of chloroquine was required if the drug was added 3 or 5 h after adsorption.

Figure 3
figure3
Timed post-infection treatment with chloroquine. This experiment is similar to that depicted in Fig. 2 except that cells were infected at 1 multiplicity of infection, and chloroquine (10, 25, and 50 μM) was added 3 or 5 h after infection.

Full size image
Ammonium chloride inhibits SARS-CoV infection of Vero E6 cells
Since chloroquine inhibited SARS-CoV infection when added before or after infection, we hypothesized that another common lysosomotropic agent, NH4Cl, might also function in a similar manner. Ammonium chloride has been widely used in studies addressing endosome-mediated virus entry. Coincidently, NH4Cl was recently shown to reduce the transduction of pseudotype viruses decorated with SARS-CoV spike protein [17, 18]. In an attempt to examine if NH4Cl functions similarly to chloroquine, we performed infection analyses in Vero E6 cells before (Fig. 4A) and after (Fig. 4B) they were treated with various concentrations of NH4Cl. In both cases, we observed a 93–99% inhibition with NH4Cl at ≥ 5 mM. These data indicated that NH4Cl (≥ 5 mM) and chloroquine (≥ 10 μM) are very effective in reducing SARS-CoV infection. These results suggest that effects of chloroquine and NH4Cl in controlling SARS CoV infection and spread might be mediated by similar mechanism(s).

Figure 4
figure4
NH 4 Cl inhibits SARS-CoV during pre or post infection treatment. NH4Cl was added to the cells either before (A) or after (B) infection, similar to what was done for chloroquine in Figs 1 and 2. Antigen-positive cells were counted, and the results were presented as in Fig. 1B.

Full size image
Effect of chloroquine and NH4Cl on cell surface expression of ACE2
We performed additional experiments to elucidate the mechanism of SARS-CoV inhibition by chloroquine and NH4Cl. Since intra-vesicular acidic pH regulates cellular functions, including N-glycosylation trimming, cellular trafficking, and various enzymatic activities, it was of interest to characterize the effect of both drugs on the processing, glycosylation, and cellular sorting of SARS-CoV spike glycoprotein and its receptor, ACE2. Flow cytometry analysis was performed on Vero E6 cells that were either untreated or treated with highly effective anti-SARS-CoV concentrations of chloroquine or NH4Cl. The results revealed that neither drug caused a significant change in the levels of cell-surface ACE2, indicating that the observed inhibitory effects on SARS-CoV infection are not due to the lack of available cell-surface ACE2 (Fig. 5A). We next analyzed the molecular forms of endogenous ACE2 in untreated Vero E6 cells and in cells that were pre-incubated for 1 h with various concentrations of either NH4Cl (2.5–10 mM) or chloroquine (1 and 10 μM) and labeled with 35S-(Met) for 3 h in the presence or absence of the drugs (Fig. 5B and 5C). Under normal conditions, we observed two immunoreactive ACE2 forms, migrating at ~105 and ~113 kDa, respectively (Fig. 5B, lane 1). The ~105-kDa protein is endoglycosidase H sensitive, suggesting that it represents the endoplasmic reticulum (ER) localized form, whereas the ~113-kDa protein is endoglycosidase H resistant and represents the Golgi-modified form of ACE2 [19]. The specificity of the antibody was confirmed by displacing the immunoreactive protein bands with excess cold-soluble human recombinant ACE2 (+ rhACE2; Fig. 5B, lane 2). When we analyzed ACE2 forms in the presence of NH4Cl, a clear stepwise increase in the migration of the ~113-kDa protein was observed with increasing concentrations of NH4Cl, with a maximal effect observed at 10 mM NH4Cl, resulting in only the ER form of ACE2 being visible on the gel (Fig. 5B, compare lanes 3–5). This suggested that the trimming and/or terminal modifications of the N-glycosylated chains of ACE2 were affected by NH4Cl treatment. In addition, at 10 mM NH4Cl, the ER form of ACE2 migrated with slightly faster mobility, indicating that NH4Cl at that concentration might also affect core glycosylation. We also examined the terminal glycosylation status of ACE2 when the cells were treated with chloroquine (Fig. 5C). Similar to NH4Cl, a stepwise increase in the electrophoretic mobility of ACE2 was observed with increasing concentrations of chloroquine. At 25 μM chloroquine, the faster electrophoretic mobility of the Golgi-modified form of ACE2 was clearly evident. On the basis of the flow cytometry and immunoprecipitation analyses, it can be inferred that NH4Cl and chloroquine both impaired the terminal glycosylation of ACE2, while NH4Cl resulted in a more dramatic effect. Although ACE2 is expressed in similar quantities at the cell surface, the variations in its glycosylation status might render the ACE2-SARS-CoV interaction less efficient and inhibit virus entry when the cells are treated with NH4Cl and chloroquine.

Figure 5
figure5
Effect of lysomotropic agents on the cell-surface expression and biosynthesis of ACE2. (A) Vero E6 cells were cultured for 20 h in the absence (control) or presence of chloroquine (10 μM) or NH4Cl (20 mM). Cells were labeled with anti-ACE2 (grey histogram) or with a secondary antibody alone (white histogram). (B) Biosynthesis of ACE2 in untreated cells or in cells treated with NH4Cl. Vero E6 cells were pulse-labeled for 3 h with 35S-Met, and the cell lysates were immunoprecipitated with an ACE2 antibody (lane 1). Preincunbation of the antibody with recombinant human ACE2 (rhACE2) completely abolished the signal (lane 2). The positions of the endoglycosidase H-sensitive ER form and the endoglycosidase H-resistant Golgi form of ACE2 are emphasized. Note that the increasing concentration of NH4Cl resulting in the decrease of the Golgi form of ACE2. (C) A similar experiment was performed in the presence of the indicated concentrations of chloroquine. Note the loss of terminal glycans with increasing concentrations of chloroquine. (D) The terminal glycosidic modification of ACE2 was evaluated by neuraminidase treatment of immunoprecipitated ACE2. Here cells were treated with 1–25 μM concentrations of chloroquine during starvation, pulse, and 3-h chase.

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To confirm that ACE2 undergoes terminal sugar modifications and that the terminal glycosylation is affected by NH4Cl or chloroquine treatment, we performed immunopreipitation of 35S-labeled ACE2 and subjected the immunoprecipitates to neuraminidase digestion. Proteins were resolved using SDS-PAGE (Fig 5D). It is evident from the slightly faster mobility of the Golgi form of ACE2 after neuraminidase treatment (Fig 5D, compare lanes 1 and 2), that ACE2 undergoes terminal glycosylation; however, the ER form of ACE2 was not affected by neuraminidase. Cells treated with 10 μM chloroquine did not result in a significant shift; whereas 25 μM chloroquine caused the Golgi form of ACE2 to resolve similar to the neuraminidase-treated ACE2 (Fig 5D, compare lanes 5 and 6). These data provide evidence that ACE2 undergoes terminal glycosylation and that chloroquine at anti-SARS-CoV concentrations abrogates the process.

Effect of chloroquine and NH4Cl on the biosynthesis and processing of SARS-CoV spike protein
We next addressed whether the lysosomotropic drugs (NH4Cl and chloroquine) affect the biosynthesis, glycosylation, and/or trafficking of the SARS-CoV spike glycoprotein. For this purpose, Vero E6 cells were infected with SARS-CoV for 18 h. Chloroquine or ammonium chloride was added to these cells during while they were being starved (1 h), labeled (30 min) or chased (3 h). The cell lysates were analyzed by immunoprecipitation with the SARS-specific polyclonal antibody (HMAF). The 30-min pulse results indicated that pro-spike (proS) was synthesized as a ~190-kDa precursor (proS-ER) and processed into ~125-, ~105-, and ~80-kDa proteins (Fig. 6A, lane 2), a result identical to that in our previous analysis [6]. Except for the 100 μM chloroquine (Fig. 6A, lane 3), there was no significant difference in the biosynthesis or processing of the virus spike protein in untreated or chloroquine-treated cells (Fig. 6A, lanes 4–6). It should be noted that chloroquine at 100 μM resulted in an overall decrease in biosynthesis and in the levels of processed virus glycoprotein. In view of the lack of reduction in the biosynthesis and processing of the spike glycoprotein in the presence of chloroquine concentrations (10 and 50 μM) that caused large reductions in SARS-CoV replication and spread, we conclude that the antiviral effect is probably not due to alteration of virus glycoprotein biosynthesis and processing. Similar analyses were performed with NH4Cl, and the data suggested that the biosynthesis and processing of the spike protein were also not negatively affected by NH4Cl (Fig. 6A, lanes 7–12). Consistent with our previous analysis [6], we observed the presence of a larger protein, which is referred to here as oligomers. Recently, Song et al. [20] provided evidence that these are homotrimers of the SARS-CoV spike protein and were incorporated into the virions. Interestingly, the levels of the homotrimers in cells treated with 100 μM chloroquine and 40 and 20 mM NH4Cl (Fig. 6A, lanes 3, 9, and 10) were slightly lower than in control cells or cells treated with lower drug concentrations.

Figure 6
figure6
Effects of NH 4 Cl and chloroquine (CQ) on the biosynthesis, processing, and glycosylation of SARS-CoV spike protein. Vero E6 cells were infected with SARS-CoV as described in Fig. 2. CQ or NH4Cl was added during the periods of starvation (1 h) and labeling (30 min) with 35S-Cys and followed by chase for 3 h in the presence of unlabeled medium. Cells were lysed in RIPA buffer and immunoprecipitated with HMAF. Virus proteins were resolved using 3–8% NuPAGE gel (Invitrogen). The cells presented were labeled for 30 min (A) and chased for 3 h (B). The migration positions of the various spike molecular forms are indicated at the right side, and those of the molecular standards are shown to the left side. proS-ER and proS-Golgi are the pro-spike of SARS-Co in the ER and Golgi compartments, respectively and proS-ungly is the unglycosylated pro-spike ER.

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The data obtained from a 30-min pulse followed by a 3-h chase (Fig. 6B, lanes 2 and Cool confirmed our earlier observation that the SARS-CoV spike protein precursor (proS-ER) acquires Golgi-specific modifications (proS-Golgi) resulting in a ~210-kDa protein [6]. Chloroquine at 10, 25, and 50 μM had no substantial negative impact on the appearance of the Golgi form (Fig. 6B, compare lane 2 to lanes 4–6). Only at 100 μM chloroquine was a reduction in the level of the Golgi-modified pro-spike observed (lane 3). On the other hand, NH4Cl abrogated the appearance of Golgi-modified forms at ≥10 mM (compare lane 8 with 9–11) and had a milder effect at 1 mM (lane 12). These data clearly demonstrate that the biosynthesis and proteolytic processing of SARS-CoV spike protein are not affected at chloroquine (25 and 50 μM) and NH4Cl (1 mM) doses that cause virus inhibitory effects. In addition, with 40, 20, and 10 mM NH4Cl, there was an increased accumulation of proS-ER with a concomitant decrease in the amount of oligomers (Fig. 6B, lanes 9–11). When we examined the homotrimers, we found that chloroquine at 100 μM and NH4Cl at 40 and 20 mM resulted in slightly faster mobility of the trimers (Fig. 6B, lanes 3, 9, and 10), but lower drug doses, which did exhibit significant antiviral effects, did not result in appreciable differences. These data suggest that the newly synthesized intracellular spike protein may not be a major target for chloroquine and NH4Cl antiviral action. The faster mobility of the trimer at certain higher concentration of the drugs might be due the effect of these drugs on the terminal glycosylation of the trimers.

Discussion
We have identified chloroquine as an effective antiviral agent for SARS-CoV in cell culture conditions, as evidenced by its inhibitory effect when the drug was added prior to infection or after the initiation and establishment of infection. The fact that chloroquine exerts an antiviral effect during pre- and post-infection conditions suggest that it is likely to have both prophylactic and therapeutic advantages. Recently, Keyaerts et al. [21] reported the antiviral properties of chloroquine and identified that the drug affects SARS-CoV replication in cell culture, as evidenced by quantitative RT-PCR. Taken together with the findings of Keyaerts et al. [21], our analysis provides further evidence that chloroquine is effective against SARS-CoV Frankfurt and Urbani strains. We have provided evidence that chloroquine is effective in preventing SARS-CoV infection in cell culture if the drug is added to the cells 24 h prior to infection. In addition, chloroquine was significantly effective even when the drug was added 3–5 h after infection, suggesting an antiviral effect even after the establishment of infection. Since similar results were obtained by NH4Cl treatment of Vero E6 cells, the underlying mechanism(s) of action of these drugs might be similar.

Apart from the probable role of chloroquine on SARS-CoV replication, the mechanisms of action of chloroquine on SARS-CoV are not fully understood. Previous studies have suggested the elevation of pH as a mechanism by which chloroquine reduces the transduction of SARS-CoV pseudotype viruses [17, 18]. We examined the effect of chloroquine and NH4Cl on the SARS-CoV spike proteins and on its receptor, ACE2. Immunoprecipitation results of ACE2 clearly demonstrated that effective anti-SARS-CoV concentrations of chloroquine and NH4Cl also impaired the terminal glycosylation of ACE2. However, the flow cytometry data demonstrated that there are no significant differences in the cell surface expression of ACE2 in cells treated with chloroquine or NH4Cl. On the basis of these results, it is reasonable to suggest that the pre-treatment with NH4Cl or chloroquine has possibly resulted in the surface expression of the under-glycosylated ACE2. In the case of chloroquine treatment prior to infection, the impairment of terminal glycosylation of ACE2 may result in reduced binding affinities between ACE2 and SARS-CoV spike protein and negatively influence the initiation of SARS-CoV infection. Since the biosynthesis, processing, Golgi modification, and oligomerization of the newly synthesized spike protein were not appreciably affected by anti-SARS-CoV concentrations of either chloroquine or NH4Cl, we conclude that these events occur in the cell independent of the presence of the drugs. The potential contribution of these drugs in the elevation of endosomal pH and its impact on subsequent virus entry or exit could not be ruled out. A decrease in SARS-CoV pseudotype transduction in the presence of NH4Cl was observed and was attributed to the effect on intracellular pH [17, 18]. When chloroquine or NH4Cl are added after infection, these agents can rapidly raise the pH and subvert on-going fusion events between virus and endosomes, thus inhibiting the infection.

In addition, the mechanism of action of NH4Cl and chloroquine might depend on when they were added to the cells. When added after the initiation of infection, these drugs might affect the endosome-mediated fusion, subsequent virus replication, or assembly and release. Previous studies of chloroquine have demonstrated that it has multiple effects on mammalian cells in addition to the elevation of endosomal pH, including the prevention of terminal glycosyaltion of immunoglobulins [22]. When added to virus-infected cells, chloroquine inhibited later stages in vesicular stomatitis virus maturation by inhibiting the glycoprotein expression at the cell surface [23], and it inhibited the production of infectious HIV-1 particles by interfering with terminal glycosylation of the glycoprotein [24, 25]. On the basis of these properties, we suggest that the cell surface expression of under-glycosylated ACE2 and its poor affinity to SARS-CoV spike protein may be the primary mechanism by which infection is prevented by drug pretreatment of cells prior to infection. On the other hand, rapid elevation of endosomal pH and abrogation of virus-endosome fusion may be the primary mechanism by which virus infection is prevented under post-treatment conditions. More detailed SARS CoV spike-ACE2 binding assays in the presence or absence of chloroquine will be performed to confirm our findings. Our studies indicate that the impact of NH4Cl and chloroquine on the ACE2 and spike protein profiles are significantly different. NH4Cl exhibits a more pronounced effect than does chloroquine on terminal glycosylation, highlighting the novel intricate differences between chloroquine and ammonium chloride in affecting the protein transport or glycosylation of SARS-CoV spike protein and its receptor, ACE2, despite their well-established similar effects of endosomal pH elevation.

The infectivity of coronaviruses other than SARS-CoV are also affected by chloroquine, as exemplified by the human CoV-229E [15]. The inhibitory effects observed on SARS-CoV infectivity and cell spread occurred in the presence of 1–10 μM chloroquine, which are plasma concentrations achievable during the prophylaxis and treatment of malaria (varying from 1.6–12.5 μM) [26] and hence are well tolerated by patients. It recently was speculated that chloroquine might be effective against SARS and the authors suggested that this compound might block the production of TNFα, IL6, or IFNγ [15]. Our data provide evidence for the possibility of using the well-established drug chloroquine in the clinical management of SARS.

Conclusion
Chloroquine, a relatively safe, effective and cheap drug used for treating many human diseases including malaria, amoebiosis and human immunodeficiency virus is effective in inhibiting the infection and spread of SARS CoV in cell culture. The fact that the drug has significant inhibitory antiviral effect when the susceptible cells were treated either prior to or after infection suggests a possible prophylactic and therapeutic use.

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