cant find sugarless moon cake
so never eat
-
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[COVID-19 Virus] The Sinkies are fucked Thread.
- Thread starter zhihau
- Start date
so never eat
In the end, he will end up talking only with himself as he has already ignored everyone else. What a loser.
The loser's path so far:
1. Began life by spreading misinformation, lies and hate to stop people from protecting themselves = Failed when 90% vaccinated showed him the finger.
2. Started a one man crusade to openly ask people to revolt against authorities so they can stop vaccination, and prevent people from protecting themselves = Failed because most antivaxxers just want to kpkb here and not do more.
3. Tried to kill people by asking them to drink Chlorine Dioxide, touting it as miracle cure = Failed because his antivaxxer pals here are not that stupid to drink cleaning agent .... at least not yet.
4. Currently asking people to commit crime by going against the law = failed because people don't want to go to jail.
Along the way, he did promise us that Trump will be reinstated as POTUS last month.
Mother-of-four who proudly posted that she was 'unmasked, unmuzzled, and unvaccainated' dies of COVID-19 at age 40
- Kristen Lowery, 40, from Escalon, California, 'unexpectedly' passed away on September 15th
- Family revealed on Facebook that she had been battling COVID-19 and resulting pneumonia ahead of her passing
- Lowery was mother to four school-age children, Tayden, McKenna, Ella, and Ryenn
- She had regularly posted anti-vax, anti-mask content, proclaiming herself a 'free thinker' who 'questions everything'
Kristen Lowery, 40, from Escalon, California, 'unexpectedly' passed away on September 15th, according to a GoFundMe page raising money for her funeral expenses.
In screenshots taken from Facebook, family members say she lost her battle against COVID-19 — after frequently posting content protesting vaccines and labeling herself 'unmasked and unmuzzled' and a 'free thinker.'
Kristen Lowery, 40, from Escalon, California, 'unexpectedly' passed away from COVID-19 on September 15th
Lowery was mother to four school-age children, Tayden, McKenna, Ella, and Ryenn
Lowery's Facebook profile has since been set to private, but screenshots have been posted on Reddit and other sites.
Lowery had made use of political borders for her profile photos, including one reading 'Unmasked, unmuzzled, unvaccainated, unafraid. Together we win.'
Another read: 'Free Thinker. Question everything. Research mandatory.'
She also appears to have gone to some sort of protest or anti-vaccine event holding a sign reading 'Give voice to the vaccine injured.' In the photo, she also wore a shirt proclaiming herself to be an 'ex-vaxxer' and reading: 'I trusted them. Never again.'
Yet another photo was lined with a border reading 'Moms for Liberty.' Lowery was mother to four school-age children, Tayden, McKenna, Ella, and Ryenn.
Lowery had made use of political borders for her profile photos, including one reading 'Unmasked, unmuzzled, unvaccainated, unafraid. Together we win'
She was outspoken on social media against vaccines and masks
But earlier this month, her sister, Cassie, wrote on Facebook that Lowery was 'in the hospital fighting for her life against Covid and pneumonia.
'My sister has a long life left. We need you here, sissy. Your kids love and miss you so much. Please don't give up,' she wrote.
She also warned that this wasn't 'political' and she was uninterested in hearing anyone's opinions on vaccines.
Then, last week, two family members posted on Facebbok that Lowery 'died from covid,' with her aunt mourning that her 'heart is breaking for her 4 kids.'
'So many people need her here, but God must have special plans for her in heaven,' her aunt wrote.
Family revealed on Facebook that she had been battling COVID-19 and resulting pneumonia ahead of her passing
On a GoFundMe page that has reaised over $11,000 for her funeral, a loved one also shared the new, which is 'devastating to everyone that knew her'
On a GoFundMe page that has reaised over $11,000 for her funeral, a loved one also shared the new, which is 'devastating to everyone that knew her.'
'Kristen was a beautiful and amazing woman but more than that she was a phenomenal mother who always, ALWAYS put her children first,' she wrote.
'She lived for her children (Tayden, McKenna, Ella, and Ryenn) and loved being their mother more than anything in her life.
'She was full of life and loved her friends and family fiercely. She was the kind of person that even if she didn't have anything she would give the shirt off of her back to anyone in need. She was everything that was good and kind.
'Her passing leaves a big hole in many of our lives and she will never be forgotten but will be greatly missed,' reads the page.
RIP dear anti-vaxxer AMDL chiobu.Mother-of-four who proudly posted that she was 'unmasked, unmuzzled, and unvaccainated' dies of COVID-19 at age 40
A California mother-of-four who was outspoken against vaccines and masks has died of COVID-19.
- Kristen Lowery, 40, from Escalon, California, 'unexpectedly' passed away on September 15th
- Family revealed on Facebook that she had been battling COVID-19 and resulting pneumonia ahead of her passing
- Lowery was mother to four school-age children, Tayden, McKenna, Ella, and Ryenn
- She had regularly posted anti-vax, anti-mask content, proclaiming herself a 'free thinker' who 'questions everything'
Kristen Lowery, 40, from Escalon, California, 'unexpectedly' passed away on September 15th, according to a GoFundMe page raising money for her funeral expenses.
In screenshots taken from Facebook, family members say she lost her battle against COVID-19 — after frequently posting content protesting vaccines and labeling herself 'unmasked and unmuzzled' and a 'free thinker.'
Kristen Lowery, 40, from Escalon, California, 'unexpectedly' passed away from COVID-19 on September 15th
Lowery was mother to four school-age children, Tayden, McKenna, Ella, and Ryenn
Lowery's Facebook profile has since been set to private, but screenshots have been posted on Reddit and other sites.
Lowery had made use of political borders for her profile photos, including one reading 'Unmasked, unmuzzled, unvaccainated, unafraid. Together we win.'
Another read: 'Free Thinker. Question everything. Research mandatory.'
She also appears to have gone to some sort of protest or anti-vaccine event holding a sign reading 'Give voice to the vaccine injured.' In the photo, she also wore a shirt proclaiming herself to be an 'ex-vaxxer' and reading: 'I trusted them. Never again.'
Yet another photo was lined with a border reading 'Moms for Liberty.' Lowery was mother to four school-age children, Tayden, McKenna, Ella, and Ryenn.
Lowery had made use of political borders for her profile photos, including one reading 'Unmasked, unmuzzled, unvaccainated, unafraid. Together we win'
She was outspoken on social media against vaccines and masks
But earlier this month, her sister, Cassie, wrote on Facebook that Lowery was 'in the hospital fighting for her life against Covid and pneumonia.
'My sister has a long life left. We need you here, sissy. Your kids love and miss you so much. Please don't give up,' she wrote.
She also warned that this wasn't 'political' and she was uninterested in hearing anyone's opinions on vaccines.
Then, last week, two family members posted on Facebbok that Lowery 'died from covid,' with her aunt mourning that her 'heart is breaking for her 4 kids.'
'So many people need her here, but God must have special plans for her in heaven,' her aunt wrote.
Family revealed on Facebook that she had been battling COVID-19 and resulting pneumonia ahead of her passing
On a GoFundMe page that has reaised over $11,000 for her funeral, a loved one also shared the new, which is 'devastating to everyone that knew her'
On a GoFundMe page that has reaised over $11,000 for her funeral, a loved one also shared the new, which is 'devastating to everyone that knew her.'
'Kristen was a beautiful and amazing woman but more than that she was a phenomenal mother who always, ALWAYS put her children first,' she wrote.
'She lived for her children (Tayden, McKenna, Ella, and Ryenn) and loved being their mother more than anything in her life.
'She was full of life and loved her friends and family fiercely. She was the kind of person that even if she didn't have anything she would give the shirt off of her back to anyone in need. She was everything that was good and kind.
'Her passing leaves a big hole in many of our lives and she will never be forgotten but will be greatly missed,' reads the page.
1173 community cases! shiok shiok or not?
Talking Donkey
Alfrescian (S)
Why you neber update today case, it's suppose to be your duty u noe?
Teacher , updated
ADVISORY ON VULNERABLE GROUP
As more information emerges about COVID-19 infections, there has been further indication of groups who are at risk of poorer clinical outcomes in the event of a COVID-19 infection.
Greater precautions should be taken for such individuals, in keeping with existing safe management measures applicable to the general population.
Such persons include:
a. Persons who are aged 60 and above;
b. Patients who are immunocompromised or have concurrent medical conditions such as obesity (BMI > 31), hypertension, diabetes, chronic heart and lung diseases, kidney diseases on dialysis, hypercoagulable states,c ancer, or patients on drugs that cause immunosuppression;
c. Pregnant women;
d. Persons with Down’s syndrome
Persons who are part of the groups identified above should take greater care to avoid exposure to COVID-19 infection, and should be encouraged to be fully vaccinated against COVID-19 if they have no medical contraindications, so as to reduce their risk of severe disease if infected.
They should pay special attention to MOH’s health advisories, to practise a high level of personal hygiene habits with frequent hand washing with soap and water. They should also comply with safe distancing and other infection prevention and control measures e.g. use of masks, avoiding crowds and avoiding close contact with others who are unwell.
We also advise them to continue to take medications for any underlying health conditions exactly as prescribed, go for regular vaccinations based on clinical indication, and to seek medical attention promptly if feeling unwell.
Employers and caregivers for at-risk persons/ groups should support them to adhere to the above advice.
MINISTRY OF HEALTH
21 SEPTEMBER 2021
still hovering around 98% not actually sick
Over the last 28 days, 97.9 per cent of the local cases were asymptomatic or had mild symptoms.
There were 246 cases who needed oxygen supplementation and 23 had been in the ICU. Of these, 54.3 per cent were fully vaccinated and 45.7 per cent were either partially vaccinated or unvaccinated.
slowly but surely, that percentage is dropping.
vax or no vax seems to make no difference to those older folks who have comorbidities
i guess thats why moh issue the advisory last night
There’s a reason why I set the mental note to look at the ICU figures in late October you know?
meanwhile, tuapayoh has becum a dangerous covid slumland...haaaa
who is the useless pap mp?
Life around Toa Payoh disrupted by Covid-19 clusters - here's what's really going on in the hood, Singapore News
www.asiaone.com
SINGAPORE - The crowds are gone in Toa Payoh, which is feeling the impact of several Covid-19 clusters emerging across the neighbourhood over the past few weeks.
The Ministry of Health has identified various locations as virus clusters, the most recent being the Singapore Pools outlet at Block 111 Toa Payoh Lorong 1 and the bus interchange in Toa Payoh Central, currently the biggest of the clusters with 254 cases.
The other clusters are the market and hawker centre at Block 210 Toa Payoh Lorong 8, the wholesale night vegetable market in Toa Payoh East Lorong 7, religious school Madrasah Irsyad Zuhri Al-Islamiah in Toa Payoh North, and Orient Goldsmiths and Jewellers in Toa Payoh Central.
The usually busy hawker centre along Toa Payoh Lorong 8 was mostly empty when The Straits Times visited on Monday (Sept 20) afternoon.
Less than a quarter of the 80 stores were in operation, with many hawkers having opted against opening on the first day since the premises had been closed for cleaning for three days.
The cleaning, initially scheduled for Friday, was brought forward to last Sunday after Covid-19 cases were detected among stallholders and workers there.
A total of 41 cases linked to the hawker centre and market have been identified.
A female hawker, who declined to be identified, said that many of the other store owners were reluctant to resume business as they were not expecting many customers to visit.
She said she had opted to open as she still had to pay for rental costs.
"Today is a really horrible day, there's no one here. During lunchtime, it's normally busy but as you can see, it's empty now," she said, pointing to rows of unoccupied tables.
Another hawker said he would usually sell 50 bowls of fishball noodles but had less than half his takings on that day.
Member of Parliament Saktiandi Supaat, who oversees the area containing both the market and hawker centre in Lorong 8 and the wholesale night vegetable market in Toa Payoh East Lorong 7, has advised residents to avoid crowds and when necessary, to make their visits brief.
"We're also encouraging our residents to perform home tests on themselves using the ART (antigen rapid test) kits," he added.
Businesses around the market were similarly affected by the news of the cluster and the weekend's closure. A supervisor at the Kelly Jie seafood restaurant, Mr Jason, 60, said that it was slim pickings for the eatery as well.
Speaking in Mandarin, he said: "We've had no customers today, just a couple of people who came by to take away their lunch. On weekends, we usually take only reservations but less than half the tables were occupied - residents staying nearby who could not go to the hawker centre as it was closed."
Over at the bus interchange in Toa Payoh Central, the usual lunchtime crowd was absent. Popular food and beverage outlets, such as Toast Box, Burger King, and McDonald's, had about half their usual number of customers.
A service staff at the Toast Box outlet, Mr Ng Yong Sun, 69, said that news of the cluster at the interchange resulted in a massive drop in the number of customers.
"On Fridays and weekends, business is generally better, but it still won't be full like it was before the cluster," he said.
Ms Easwar Utamrao Soorin, 29, a sales assistant at the Fashion One clothing shop, reported a similar scenario. She said: "At 9am when I opened the shop, there was no one at all. Some people came only after lunch."
The cluster was first reported on Aug 26 when proactive testing by the respective public transport operators picked up 36 cases linked to various bus interchanges.
A stall worker sits at a table in the market and hawker centre at Block 210 Toa Payoh Lorong 8, on Sept 20. PHOTO: Straits Times/Wallace Woon
According to Transport Minister Mr S. Iswaran, less than 10 per cent of the 9,500 public bus drivers are affected by the outbreak at the interchanges.
Even though he noted that the impact on bus services had been limited, the lack of manpower has seen commuters facing longer wait times for bus services.
"I usually wait ten minutes for the bus, but now it has doubled to 20 minutes," said Mr Soh Hun Seng, 68.
"I try to stay home as much as possible, but today I went out to meet my friend for lunch as I felt cooped up at home."
Housewife Madam Sariyah Nacknay, 43, who was on the way home from school with her daughter, who is in Primary 3, said that there was a five-minute longer wait time for the bus, compared to normal.
Taking it in her stride, she said: "It doesn't matter to me because I don't like to rush anyway. I can see the bus uncles are making an effort to go to work, drive us around, so I appreciate that."
A man walks past shuttered food stalls in the market and hawker centre at Block 210 Toa Payoh Lorong 8, on Sept 20. PHOTO: Straits Times/Wallace Woon
The Orient Goldsmith and Jewellery was first designated a cluster on Sept 4, and grew to a total of 17 cases linked to the location. As of Monday, the store has remained closed since Sept 3.
Neighbouring shops along the stretch reported a similar downturn in their business. Many shopkeepers could be seen sitting in their stores, customers nowhere to be found.
A few units down at Newhouse Furnishing, store boss Mr Johnny Chan, 60 said that the dwindling footfall could sound the death knell for his business.
"My business is dying. Our business has dropped by 60 per cent - I don't have enough to pay rent. In fact, business has been dropping since around May or June.
917 new Covid-19 cases in Singapore; 2 Singaporean men aged 84 and 85 die from complications
"By the end of the year, if business doesn't improve, we can't hang on any longer and I'll have to close down my shop. I can't even say I'm sad because the Government has done what it can, and other businesses like the travel agencies and karaoke places have it worse than me," he added.
For some residents, the emergence of the clusters comes as no surprise.
"The people here don't care about Covid, especially the old people. They just sit around and mingle, they drink and chit chat," said Mr Stanley Quek, 68, a transport driver for foreign workers.
Echoing his sentiments was Madam Ho, 60, who works in a bakery next to the closed Singapore Pools outlet at Block 111 along Toa Payoh Lorong 1.
The betting outlet was closed after an employee there tested positive for Covid-19 on Friday.
An elderly man reads a closure notice of the Singapore Pools outlet at Block 111 Toa Payoh Lorong 1, on Sept 20. PHOTO: Straits Times/Wallace Woon
Members of the public could be seen closely reading a notice on the shutter informing them of the closure, advising them to patronise alternate outlets in the area.
She said: "The queue next door (at the Singapore Pools) has no safe-distancing and people don't wear their masks properly all the time."
Construction supervisor Mr Kelvin Yeo, 62, said that the pandemic has just become a "part of life".
"The Covid-19 situation didn't really affect my daily life because I never liked to go to crowded places to begin with," he said.
"There are new clusters everyday and we cannot keep living in fear. We cannot cover all the bases completely to prevent the cases from happening or rising."
who is the useless pap mp?
Life around Toa Payoh disrupted by Covid-19 clusters - here's what's really going on in the hood, Singapore News
www.asiaone.com
The Ministry of Health has identified various locations as virus clusters, the most recent being the Singapore Pools outlet at Block 111 Toa Payoh Lorong 1 and the bus interchange in Toa Payoh Central, currently the biggest of the clusters with 254 cases.
The other clusters are the market and hawker centre at Block 210 Toa Payoh Lorong 8, the wholesale night vegetable market in Toa Payoh East Lorong 7, religious school Madrasah Irsyad Zuhri Al-Islamiah in Toa Payoh North, and Orient Goldsmiths and Jewellers in Toa Payoh Central.
The usually busy hawker centre along Toa Payoh Lorong 8 was mostly empty when The Straits Times visited on Monday (Sept 20) afternoon.
Less than a quarter of the 80 stores were in operation, with many hawkers having opted against opening on the first day since the premises had been closed for cleaning for three days.
The cleaning, initially scheduled for Friday, was brought forward to last Sunday after Covid-19 cases were detected among stallholders and workers there.
A total of 41 cases linked to the hawker centre and market have been identified.
A female hawker, who declined to be identified, said that many of the other store owners were reluctant to resume business as they were not expecting many customers to visit.
She said she had opted to open as she still had to pay for rental costs.
"Today is a really horrible day, there's no one here. During lunchtime, it's normally busy but as you can see, it's empty now," she said, pointing to rows of unoccupied tables.
Another hawker said he would usually sell 50 bowls of fishball noodles but had less than half his takings on that day.
Member of Parliament Saktiandi Supaat, who oversees the area containing both the market and hawker centre in Lorong 8 and the wholesale night vegetable market in Toa Payoh East Lorong 7, has advised residents to avoid crowds and when necessary, to make their visits brief.
"We're also encouraging our residents to perform home tests on themselves using the ART (antigen rapid test) kits," he added.
Businesses around the market were similarly affected by the news of the cluster and the weekend's closure. A supervisor at the Kelly Jie seafood restaurant, Mr Jason, 60, said that it was slim pickings for the eatery as well.
Speaking in Mandarin, he said: "We've had no customers today, just a couple of people who came by to take away their lunch. On weekends, we usually take only reservations but less than half the tables were occupied - residents staying nearby who could not go to the hawker centre as it was closed."
Over at the bus interchange in Toa Payoh Central, the usual lunchtime crowd was absent. Popular food and beverage outlets, such as Toast Box, Burger King, and McDonald's, had about half their usual number of customers.
A service staff at the Toast Box outlet, Mr Ng Yong Sun, 69, said that news of the cluster at the interchange resulted in a massive drop in the number of customers.
"On Fridays and weekends, business is generally better, but it still won't be full like it was before the cluster," he said.
Ms Easwar Utamrao Soorin, 29, a sales assistant at the Fashion One clothing shop, reported a similar scenario. She said: "At 9am when I opened the shop, there was no one at all. Some people came only after lunch."
The cluster was first reported on Aug 26 when proactive testing by the respective public transport operators picked up 36 cases linked to various bus interchanges.
According to Transport Minister Mr S. Iswaran, less than 10 per cent of the 9,500 public bus drivers are affected by the outbreak at the interchanges.
Even though he noted that the impact on bus services had been limited, the lack of manpower has seen commuters facing longer wait times for bus services.
"I usually wait ten minutes for the bus, but now it has doubled to 20 minutes," said Mr Soh Hun Seng, 68.
"I try to stay home as much as possible, but today I went out to meet my friend for lunch as I felt cooped up at home."
Housewife Madam Sariyah Nacknay, 43, who was on the way home from school with her daughter, who is in Primary 3, said that there was a five-minute longer wait time for the bus, compared to normal.
Taking it in her stride, she said: "It doesn't matter to me because I don't like to rush anyway. I can see the bus uncles are making an effort to go to work, drive us around, so I appreciate that."
The Orient Goldsmith and Jewellery was first designated a cluster on Sept 4, and grew to a total of 17 cases linked to the location. As of Monday, the store has remained closed since Sept 3.
Neighbouring shops along the stretch reported a similar downturn in their business. Many shopkeepers could be seen sitting in their stores, customers nowhere to be found.
A few units down at Newhouse Furnishing, store boss Mr Johnny Chan, 60 said that the dwindling footfall could sound the death knell for his business.
"My business is dying. Our business has dropped by 60 per cent - I don't have enough to pay rent. In fact, business has been dropping since around May or June.
917 new Covid-19 cases in Singapore; 2 Singaporean men aged 84 and 85 die from complications
"By the end of the year, if business doesn't improve, we can't hang on any longer and I'll have to close down my shop. I can't even say I'm sad because the Government has done what it can, and other businesses like the travel agencies and karaoke places have it worse than me," he added.
For some residents, the emergence of the clusters comes as no surprise.
"The people here don't care about Covid, especially the old people. They just sit around and mingle, they drink and chit chat," said Mr Stanley Quek, 68, a transport driver for foreign workers.
Echoing his sentiments was Madam Ho, 60, who works in a bakery next to the closed Singapore Pools outlet at Block 111 along Toa Payoh Lorong 1.
The betting outlet was closed after an employee there tested positive for Covid-19 on Friday.
Members of the public could be seen closely reading a notice on the shutter informing them of the closure, advising them to patronise alternate outlets in the area.
She said: "The queue next door (at the Singapore Pools) has no safe-distancing and people don't wear their masks properly all the time."
Construction supervisor Mr Kelvin Yeo, 62, said that the pandemic has just become a "part of life".
"The Covid-19 situation didn't really affect my daily life because I never liked to go to crowded places to begin with," he said.
"There are new clusters everyday and we cannot keep living in fear. We cannot cover all the bases completely to prevent the cases from happening or rising."
sars claimed 33 lives in sinkypura
covid has now doubled that amount and increasing
scared or not?
whether vax or unvax, if u have hypertension and hyperlipidaemia, you are at high risk. obese people beware!
Case 66111, a 62 year-old female Singaporean, has passed away from complications due to COVID-19 infection on 20 September 2021. She tested positive for COVID-19 infection on 23 July and was admitted to Khoo Teck Puat Hospital the next day. She had not been vaccinated against COVID-19, and had a history of hypertension and hyperlipidaemia.
Case 72610, a 74 year-old male Singaporean, has passed away from complications due to COVID-19 infection on 19 September 2021. He was admitted to Tan Tock Seng Hospital on 31 August for an unrelated medical condition and tested negative for COVID-19 upon admission. Another test taken on 9 September came back positive for COVID-19 infection. He had been partially vaccinated against COVID-19, and had a history of chronic kidney disease, diabetes mellitus, hypertension and hyperlipidaemia.
Case 76425, an 83 year-old male Singaporean, has passed away from complications due to COVID-19 infection on 20 September 2021. He was conveyed to Ng Teng Fong General Hospital on 15 September with symptoms, and tested positive for COVID-19 infection on the same day. He had been vaccinated against COVID-19, and had a history of chronic obstructive pulmonary disease, recurrent aspiration pneumonia, hypertension and hyperlipidaemia which, together with his advanced age, made him more susceptible to severe illness.
Does cholesterol play a role in COVID-19?
www.news-medical.net
By Dr. Liji Thomas, MDMay 12 2020
A new study published on the preprint server bioRxiv* in May 2020 suggests that the strikingly increased mortality in COVID-19 patients who are either old or also have high blood pressure, diabetes, cardiovascular disease is due to high tissue cholesterol levels. It also indicates the role of a healthy diet, by showing that polyunsaturated fatty acids (PUFA) in blood oppose the effects of cholesterol and thus the risk for severe COVID-19 symptoms.
The current study aims to answer the question as to what controls the entry of SARS-CoV-2 into the host cell.
These GM-1 rafts migrate from areas of disordered lipids or polyunsaturated fats.
Cholesterol is a fat found in cell membranes all over the body and deposited in macrophage-rich tissues with chronic inflammation. In lung disease, lung cells, as well as macrophages, are observed to take cholesterol in and out. The study, therefore, focused on finding the relationship between the infective potential of the virus and the concentration of cholesterol.
In the cholesterol loading condition, the viral entry increased markedly by 50%, while it was significantly reduced in the unloading state. The addition of a cholesterol-removing chemical called methyl-beta-cyclodextrin (MβCD) to the cell culture duplicated the viral entry-inhibiting effect of cholesterol unloading. The researchers concluded that the lack of virus entry with the unloading of cholesterol is not due to a lack of ACE2 receptor, because the latter was expressed at normal high levels in the presence of both apoE and MβCD.
Having indicated the cholesterol-dependent entry of the virus, they examined the underlying molecular mechanisms. Cholesterol is essential to the number and size of GM1 rafts as well as the in and out movement of proteins into the rafts in the cell membrane. With an increase in these, the viral entry also goes up.
The researchers found that ACE2 receptors were correlated with GM1 lipid rafts more than 3-fold in the loaded condition of the cell. The opposite effect (an over 70% reduction in localization to the rafts) was seen with the addition of MβCD. Thus apoE-mediated cholesterol loading causes localization of ACE2 at the point of viral entry.
The researchers next attempted to find out whether the reduced viral entry was due to less receptor binding. They tested the binding of the virus to the receptor-binding domain (RBD) of the ACE2 receptor on the culture cells, both with and without the addition of MβCD to the cells. The slight reduction of the RBD binding due to the presence of the chemical was insignificant.
The next step was to find out how viral entry was affected by ACE2 expression and low cholesterol levels. They found that with higher ACE2 concentrations, the receptors moved away from the vicinity of the GM1 lipid rafts, possibly because the excess ACE2 migrated to the disordered lipid region. Though the virus attaches to the ACE2 in this region, it cannot enter the cell, and thus this state protects against infection, as seen in children.
In the pseudovirus, this site is cleaved in the cells that manufacture the virus. On the other hand, another site cleaves a fusion peptide and allows the virus to enter the host cell membrane. This is supposed to be cleaved by the proteases TMPRSS2 or cathepsin.
To test this, they labeled both furin and TMPRSS2 along with the GM1 rafts. They found only furin was associated with the rafts. In low cholesterol conditions, this association disappeared.
Finally, the study also reveals the attachment of palmitoyl residues during post-translation modification. These 16-carbon lipids attach to many proteins to facilitate their entry and exit from GM1 lipid rafts. The SARS-CoV endodomain must be palmitoylated to allow viral fusion and entry via the S protein. All the palmitoylation sites considered to be present in this virus are also found in SARS-CoV-2, with one more, which may indicate that this alteration favors virus survival.
Overall, therefore, the study suggests 3 distinct mechanisms that could explain the differences in infectivity between population subgroups with comorbidities and chronic inflammation.
The first is the cholesterol-dependent increase in the number and size of GM1 lipid rafts, and hence entry points for the virus. Secondly, the receptor can dock into the sites that offer the most significant benefit for viral entry, again in a cholesterol-dependent manner. And thirdly, the proteases required for virus activation prefusion are also absent in low cholesterol conditions.
In children, who typically have surface viral entry only rather than lipid-raft-mediated endocytosis, the limited viral entry leads to mild infection. Low cholesterol reduces the infective potential of the virus still more.
With higher cholesterol, RBD binding improves, which could indicate the preferential use of endocytosis by the virus for cell entry. The increase in TMPRSS2 is linked to higher infectivity. However, if this protease is preferred in children, it might be ineffective in older and sicker patients.
One key example of this seeming paradox is the presence of chronic inflammation, where tissue cholesterol levels are high though blood cholesterol is low. The reason is that cholesterol unloading in the tissue is inhibited, while macrophages load it into the cells.
The study could thus help forward research into the role of cholesterol in viral infection in COVID-19. More research is needed to find the role of other proteins and lipids in the complex membrane environment, which could significantly influence the virus entry and infection of the host cell.
www.news-medical.net
A new study published on the preprint server bioRxiv* in May 2020 suggests that the strikingly increased mortality in COVID-19 patients who are either old or also have high blood pressure, diabetes, cardiovascular disease is due to high tissue cholesterol levels. It also indicates the role of a healthy diet, by showing that polyunsaturated fatty acids (PUFA) in blood oppose the effects of cholesterol and thus the risk for severe COVID-19 symptoms.
Variation in Infectivity Between Adults and Children
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes COVID-19 disease, is known to infect and enter host cells employing the ACE2 receptor molecule present on the outside of the host cell membrane. It was initially thought that low ACE2 levels in children caused the decreased risk for this condition, but recent studies show that this is not necessarily always true. Some children show high ACE2 levels in early childhood.The current study aims to answer the question as to what controls the entry of SARS-CoV-2 into the host cell.
The Role of Cholesterol in Viral Entry
Earlier SARS-CoV viral research might help provide the answer. This virus enters the cell only in the presence of cholesterol and monosialotetrahexosylganglioside1 (GM1) in the form of lipid rafts, in culture. Cholesterol is required for the formation of these GM-1 lipid rafts, and viruses that attach here enter the cell by endocytosis.These GM-1 rafts migrate from areas of disordered lipids or polyunsaturated fats.
Cholesterol is a fat found in cell membranes all over the body and deposited in macrophage-rich tissues with chronic inflammation. In lung disease, lung cells, as well as macrophages, are observed to take cholesterol in and out. The study, therefore, focused on finding the relationship between the infective potential of the virus and the concentration of cholesterol.
Cholesterol Promotes Viral Entry
First, using a pseudovirus expressing the spike (S) protein of SARS-CoV-2, they loaded cholesterol into cultured cells along with the carrier protein apoE, and blood serum. The S protein is responsible for viral entry. In the cholesterol-bound form, the apoE protein binds to the LDL receptor of the cell to load cholesterol into the cell. If free, its binding promotes the unloading of cholesterol from the cell.In the cholesterol loading condition, the viral entry increased markedly by 50%, while it was significantly reduced in the unloading state. The addition of a cholesterol-removing chemical called methyl-beta-cyclodextrin (MβCD) to the cell culture duplicated the viral entry-inhibiting effect of cholesterol unloading. The researchers concluded that the lack of virus entry with the unloading of cholesterol is not due to a lack of ACE2 receptor, because the latter was expressed at normal high levels in the presence of both apoE and MβCD.
Having indicated the cholesterol-dependent entry of the virus, they examined the underlying molecular mechanisms. Cholesterol is essential to the number and size of GM1 rafts as well as the in and out movement of proteins into the rafts in the cell membrane. With an increase in these, the viral entry also goes up.
Cholesterol Facilitates Greater ACE2 Availability
The movement of ACE2 receptors to the viral entry site also increases with the cholesterol load. To find out how much of the receptor is localized in these raft sites, and how much this varies with the cholesterol concentration, they used fluorescent-labeled antibodies to ACE2 to cells treated with apoE and serum and used dSTORM (direct stochastical optical reconstruction microscopy) to acquire images of the lipid domain structure at the nanoscale.The researchers found that ACE2 receptors were correlated with GM1 lipid rafts more than 3-fold in the loaded condition of the cell. The opposite effect (an over 70% reduction in localization to the rafts) was seen with the addition of MβCD. Thus apoE-mediated cholesterol loading causes localization of ACE2 at the point of viral entry.
Why is COVID-19 So Much More Dangerous in the Old and Sick?
This cholesterol-dependent mechanism of ACE2 localization, as well as virus entry, could be why COVID-19 is so much more lethal in the elderly. Tissue cholesterol levels increase with age as a result of chronic disease conditions like atherosclerosis and other acute or chronic inflammations. These are the same conditions that are found to increase the severity and likelihood of symptomatic COVID-19 disease.Related Stories
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The researchers next attempted to find out whether the reduced viral entry was due to less receptor binding. They tested the binding of the virus to the receptor-binding domain (RBD) of the ACE2 receptor on the culture cells, both with and without the addition of MβCD to the cells. The slight reduction of the RBD binding due to the presence of the chemical was insignificant.
The next step was to find out how viral entry was affected by ACE2 expression and low cholesterol levels. They found that with higher ACE2 concentrations, the receptors moved away from the vicinity of the GM1 lipid rafts, possibly because the excess ACE2 migrated to the disordered lipid region. Though the virus attaches to the ACE2 in this region, it cannot enter the cell, and thus this state protects against infection, as seen in children.
Cholesterol Increases Furin Availability
The virus requires protease-mediated priming to enter the host cell. In SARS-CoV-2, this is performed by furin, which has a cleavage site, unique to this virus, between S1 and S2 subunits of the S protein.In the pseudovirus, this site is cleaved in the cells that manufacture the virus. On the other hand, another site cleaves a fusion peptide and allows the virus to enter the host cell membrane. This is supposed to be cleaved by the proteases TMPRSS2 or cathepsin.
To test this, they labeled both furin and TMPRSS2 along with the GM1 rafts. They found only furin was associated with the rafts. In low cholesterol conditions, this association disappeared.
Finally, the study also reveals the attachment of palmitoyl residues during post-translation modification. These 16-carbon lipids attach to many proteins to facilitate their entry and exit from GM1 lipid rafts. The SARS-CoV endodomain must be palmitoylated to allow viral fusion and entry via the S protein. All the palmitoylation sites considered to be present in this virus are also found in SARS-CoV-2, with one more, which may indicate that this alteration favors virus survival.
Multiple Mechanisms of Cholesterol-Dependent Infectivity
The model shows that in high cholesterol regions, more and larger GM1 lipid rafts are formed in the cell membrane due to cholesterol loading. ACE2 moves into the GM1 lipid raft and enters the cell via endocytosis. The spike protein covering the virus surface binds to ACE2, possibly pulling the GM1 lipids over the virus and so facilitating endosomal entry.Overall, therefore, the study suggests 3 distinct mechanisms that could explain the differences in infectivity between population subgroups with comorbidities and chronic inflammation.
The first is the cholesterol-dependent increase in the number and size of GM1 lipid rafts, and hence entry points for the virus. Secondly, the receptor can dock into the sites that offer the most significant benefit for viral entry, again in a cholesterol-dependent manner. And thirdly, the proteases required for virus activation prefusion are also absent in low cholesterol conditions.
In children, who typically have surface viral entry only rather than lipid-raft-mediated endocytosis, the limited viral entry leads to mild infection. Low cholesterol reduces the infective potential of the virus still more.
With higher cholesterol, RBD binding improves, which could indicate the preferential use of endocytosis by the virus for cell entry. The increase in TMPRSS2 is linked to higher infectivity. However, if this protease is preferred in children, it might be ineffective in older and sicker patients.
How is This Important To COVID-19 Treatment?
The fact that most cell cultures for SARS-CoV-2 are carried out without a cholesterol source like serum is vital in developing therapies for COVID-19, in the light of these findings. It is vital to note that a high tissue level of cholesterol and not a high blood level is the driver of infectivity with this virus. This means that current tests for blood cholesterol do not always indicate the actual risk of severe or critical disease.One key example of this seeming paradox is the presence of chronic inflammation, where tissue cholesterol levels are high though blood cholesterol is low. The reason is that cholesterol unloading in the tissue is inhibited, while macrophages load it into the cells.
The study could thus help forward research into the role of cholesterol in viral infection in COVID-19. More research is needed to find the role of other proteins and lipids in the complex membrane environment, which could significantly influence the virus entry and infection of the host cell.
the cai png song for a break from cv19
[Sent by Gov.sg – 22 Sep]
As of 21 Sep 2021, 12pm, 1,109 COVID-19 cases are warded in hospital. There are 147 cases of serious illness requiring oxygen supplementation and 17 in the ICU.
Over the last 28 days, of the infected individuals, 97.9% have mild or no symptoms, 1.9% requires oxygen supplementation, 0.2% requires ICU care, and 0.06% has died.
As of 20 Sep, 82% of our population has completed their full regimen/received 2 doses of vaccines, and 84% has received at least one dose.
As of 21 Sep, there are 1,178 new cases in Singapore.
go.gov.sg/moh210921
As of 21 Sep 2021, 12pm, 1,109 COVID-19 cases are warded in hospital. There are 147 cases of serious illness requiring oxygen supplementation and 17 in the ICU.
Over the last 28 days, of the infected individuals, 97.9% have mild or no symptoms, 1.9% requires oxygen supplementation, 0.2% requires ICU care, and 0.06% has died.
As of 20 Sep, 82% of our population has completed their full regimen/received 2 doses of vaccines, and 84% has received at least one dose.
As of 21 Sep, there are 1,178 new cases in Singapore.
go.gov.sg/moh210921
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