BC SCIENCE
Saved by @Daoyu15

https://t.co/hXlo8qgkD0
Look like that they got a classical case of PCR Cross-Contamination.
They had 2 fabricated samples (SRX9714436 and SRX9714921) on the same PCR run. Alongside with Lung07. They did not perform metagenomic sequencing on the “feces” and they did not get

 A positive oral or anal swab from anywhere in their sampling. Feces came from anus and if these were positive the anal swabs must also be positive. Clearly it got there after the NA have been extracted and were from the very low-level degraded RNA which were mutagenized from
The Taq. https://t.co/yKXCgiT29w to see SRX9714921 and SRX9714436.
Human+Mouse in the positive SRA, human in both of them. Seeing human+mouse in identical proportions across 3 different sequencers (PRJNA573298, A22, SEX9714436) are pretty straight indication that the originals
Were already contaminated with Human and mouse from the very beginning, and that this contamination is due to dishonesty in the sample handling process which prescribe a spiking of samples in ACE2-HEK293T/A549, VERO E6 and Human lung xenograft mouse.
The “lineages” they claimed to have found aren’t mutational lineages at all—all the mutations they see on these sequences were unique to that specific sequence, and are the result of RNA degradation and from the Taq polymerase errors accumulated from the nested PCR process
Where the extremely low error-ridden cross-contaminating molecule were amplified with mutagenesis happening on each step—especially between the initial RT-PCR and sequencing PCR process, which cross contamination have happened. Notice that any extra Pro or Cys on the highly
Conserved N protein sequences are lethal to the virus—especially the extra Cys which will become a nucleophile and a Ribonuclease in the budded virion in an extracellular environment, degrading the genome and inactivating the virus immediately. These Cys will also cause the N to
Dimerize and consequently lose it’s ability to bind RNA or mediate virion assembly in the first place. While the Pro will break a conserved secondary structure in the C-terminal domain and prevent the N from folding correctly. These are not found anywhere else and are obviously
Sequencing errors. Especially since these samples came from cross-contamination without a corresponding positive oral or anal swab (they didn’t get a single swab that is positive).
M1, they cite as having “hemorrhage” claimed to be Coronavirus symptoms, are anything BUT coronavirus symptoms. https://t.co/3a8nqY6rOz “Open in new tab
X-ray tests revealed a large area of shadow in the left lung of pangolin 2-Lishui, suggestive of pneumonia” but
“we have not found coronaviruses in these pangolins by meta-transcriptomic and PCR methods.”. Notice the PLT- in the Pestivirus-infected pangolin samples. Pestivirus is seen in the SRA of M1, SRX7756769 (107267359 spots as in the Xiao et al article). The Pestivirus is seen
In all PRJNA573298 sample lungs, and it causes interstitial pneumonia and gives the same kind of CT scan and blood test result as in the 06/22 article. This sample also contained a locally circulating CCoV (Canine Coronavirus, often infects zoo/rescued solitary animals due to
It’s ubiquitous nature in most human-associated environments) strain with reads that weren’t rolling circles as the GD-1 reads and are mostly related to the Canine Coronavirus isolates circulating in dogs in GZ.
Coronavirus infections can not generate rolling circles as they does not encode RNA ligases, an enzyme that is necessary to create a true CircRNA. (What they claim to be CircRNA reads from CoV infections in other papers aren’t true CircRNA but are Partially duplicated RNA due to
The Coronavirus RdRp template-switching to a location before it’s current location. This RNA is still linear. However, the Rolling Circles corresponds to two hotspots of M13 primers seen in the amplicons for nsp3 and for the 3’-end of the genome. This imply that they were
Remnants of cDNA cloning, stemming from DNA that were circularized without a vector backbone.)
Canine Coronavirus and FIPV (alphacoronavirus 1) infections also generate very similar symptoms, and will result in positive cross-reactive NP in the immunofluorescence screening. Even the right GEO profile provide they remove it from the final SRA. Finally, the ACE2 expression
“Lowest in the lungs” claimed in this paper is at odd with the 06/22 paper claim that “ACE2 and TMPRSS2 is most highly expressed in the lungs”. This suggest that the GEO profile of the 06/22 article associated with A22/SRX8582289 have been fabricated or were derived from hACE2
Mouse. The hemorrhage seen in M1 is consistent with DYPV(Flavivirus/pestivirus related to BVDV) which is seen in the metagenome, while the pneumonia could be either be the result of DYPV-related Pestivirus, from the Sendai (murine respirovirus), from the Canine Coronavirus or
From any of the non-Coronavirus (Colitivirus or Murine Respirovirus) agents found in PRJNA573298.
M1 have Murine Respirovirus.
https://t.co/ZJnLSxwe5o
SeV is in the same family as human Parainfluenza virus. It causes bilateral ground-glass opacity.
https://t.co/NeY2ajwsDo
The blood tests—pestivirus once again fully explains all the blood changed observed. When a Parainfluenza virus is also present, the blood test become indistinguishable.
Another snRNA-seq say ACE2 and TMPRSS2 are barely exiressed in the lungs and are never coexpressed.
Weaker than the kidneys—this is again at odds with the SCAU claim that even negative pangolins have lung>>kidney for ACE2. Both publication say that Kidney is the highest ACE2 expressing organ in all pangolins where lung expression is negligible. The 06/22 article is the only
One that claim that all pangolins including negative samples, have lung ACE2 >> Kidney. This indicate that they have Spiked all of them with K12-hACE2 mouse as it is in disagreement of all previous or later pangolin transcriptomes.
“(Individual number: L08) as a positive control”—and guess what L08 actually is—Human AND Mice!
L08: Lung08.

More from Daoyu

Daoyu
Daoyu
@Daoyu15
@stevenemassey Only a sample is there. This cell line don’t have NGS. But it was consistent with a cell culture, and the RBD of the virus is not suitable for binding to the ACE2 found there. The ACE2 is same as Ra4331 and Ra9479.

And the RBD have a T403 that prevents infection of intestinal cells (hACE2 intestinal organoids, infection is restored upon T403R, but weaker than SARS-CoV-2) or lung cells (R.affinis primary, with supplemented hACE2. Infection is restored upon T403R).


Also that there exist no Sarbecovirus RBD with a D501.
ALL
Daoyu
Daoyu
@Daoyu15
@jbloom_lab It seems that it is a deeply cultured, but in deed ancestrally derived, sequence of SARS-CoV-2, that have many of the VERO cell specific mutations and deletions already in.

but it is not likely a clinical specimen from China because of the USA/WA-1 base which is not in Wuhan.

It had T22657C, T3346C, A21562C and G487T. all of which is in RaTg13 but not in WuHu-1. also T1963C and T22963C in BANAL-52. not all of the reads were H655Y and del I68-T76.

And it is actually clustered closely to the A/North America lineage, which shouldn't have appeared in a Chinese lab. also not there in Wuhan either.

also only one of the SRRs harbored T18060C it seemed.
ALL
Advertisement
Daoyu
Daoyu
@Daoyu15
@franciscodeasis https://t.co/Sd9IslUCH5 a FCS need a FCS in the inoculum to exist. It can not arise de-novo as it will be destroyed instantly by the immune system.

https://t.co/UgXygDjYbW a fourth Sars-like CoV is live at the WIV. This fourth virus is an infectious clone, where engineering of the S1-S2 is used regularly as mean to generate a culturable virus in HAE cells. No VERO E6 here, and HeLa-hACE2 is the new VERO of the WIV

https://t.co/DtjyycKy1v
https://t.co/PG7LVnHfsy
Even with VERO E6, only half the time does passage lead to the loss of the FCS—smaller plaques need to be explicitly picked for that to be a certainty.

Marburg virus is a novel virus that escaped from the lab. https://t.co/OGQM6qV27l the only reason why it did not become a pandemic is due to it being too lethal to sustain asymptomatic transmission in humans.

The highest reported number of cases were in WuChang right on top of the old WIV headquarters, In contrast to the population density data of Wuhan—note that the place near the market had the highest population density in all of Wuhan, which make it the most optimal location for
ALL
Daoyu
Daoyu
@Daoyu15
@AngloScot2 @WisdomRebel https://t.co/iHKwUoNLJ5

https://t.co/sxm17Eu9hV

https://t.co/FtjifSGItc
PRRA is highly purified in the CaLu-3 cell line when FBS is added to inhibit trypsin. Stu lied about that in his defense on the FCS identity. https://t.co/GzM7uA6vup the Gallaher article contain glaring

@WisdomRebel Mistakes and is completely impossible as HKU9 + RaTG13 only lead to a frameshift inactivated S and not a furin cleaved S.
https://t.co/gmDXK9OluN
The serological test the WIV claimed to be negative in Shi’s addendum is the exact same test they claimed positive in 2012 in the phD

@WisdomRebel thesis, and Stu once again lied about it. https://t.co/Uiy8U9BTYp significant evidence in the form of contaminated SRA datasets suggest that the DEFUSE grant have been funded in China, as the full-length HKU4 is not a part of the 2016-2019 grant, but is found before the start

@WisdomRebel Date of the 2020-2024 grant of June 2020. The WIV also contained several HKU3-like Coronaviruses, particularly in mice in an 2017 GEO experiment, which is consistent with the published HKU3 and batified mice experiment in the DEFUSE document (evidence from an already conducted

@WisdomRebel Experiment). Either DEFUSE or fast-tracked form of the 2020-2024 grant in 2019 would have lead to full-length Sarbecovirus clones being used, leading to SARS-CoV-2 in the lab—they used HKU4 in stead of HKU5 for the MERS spike experiment indicating that backbone sequence distance
ALL
Daoyu
Daoyu
@Daoyu15
https://t.co/SvP0tfbDki


https://t.co/k9mmL9e5kn


https://t.co/7fR13yb3qJ


https://t.co/EXerlaiG9u


https://t.co/iNCdLY1Pzn
ALL

More from Science

Billy Bostickson \U0001f3f4\U0001f441&\U0001f441 \U0001f193
Billy Bostickson 🏴👁&👁 ...
@BillyBostickson
Let Uncle Ralph educate you
Synthetic Viral Genomics:
Risks and Benefits for Science and Society

"Alternatively, “No See’m” sites can be used to insert foreign genes into viral, eukaryotic, or microbial genome or vector, simultaneously removing all evidence of the restriction sites that were used in the recombinant DNA manipulation"

By orientating the restriction sites as “No See’m”, the sites are removed during reassembly, leaving only the desired mutation in the final DNA product.

The dual properties of strand specificity & a variable end overhang that can be tailored to match any sequence allow for Esp3I sites to be engineered as “universal connectors” that can be joined with any other four nucleotide restriction site overhangs (e.g. EcoRI, PstX1, BamH1)

Seamless assembly (also called No See’m Sites (85)) cascades have been used to assemble full length cDNAs of the coronaviruses mouse hepatitis virus, transmissible gastroenteritis virus, infectious bronchitis virus and SARS-CoV (Refs: 85,86,87)
SCIENCE , ALL
Marit Kolby Zin\xf6cker
Marit Kolby Zinöcker...
@zinocker
Did we get dietary saturated fats all wrong? The #HADLmodel provides a new understanding and an opportunity to get it right. THREAD👇👇👇
@simondankel @kariannesve

Increased dietary saturated fatty acids lead to increased cholesterol in lipoproteins, but we don’t know why. Enter the #HADLmodel, which explains changes in lipoprotein cholesterol as adaptive homeostatic adjustments that ensure optimal cell membrane fluidity and cell function.

We propose that circulating lipoproteins enable appropriate redistribution of cholesterol molecules between specific cells and tissues, to accomodate changes in dietary fatty acid supply, due to our omnivore nature and variable intake of fatty acids. #HADLmodel

Our #HADLmodel implies that circulating levels of LDL change for protective, not for pathological reasons; an SFA-induced raise in LDL cholesterol in healthy individuals is a normal response, while a lack of this needed response may reflect a deeper pathology in lipid handling.

Circulating lipoproteins may change for pathological reasons, when regulatory mechanisms become disrupted by pathogenic processes related e.g. to inflammatory processes. Diverging lipoprotein responses in healthy versus metabolically unhealthy individuals support this view.
SCIENCE
Advertisement
The Tycoon Mindset
The Tycoon Mindset
@tycoonmindset05
Solara Active Pharma Science con call was today at 3:30 PM.

@Gautam__Baid @unseenvalue @drprashantmish6 @rdkriplani

Here are the key takeaways of the call. 👇🧵

Business Updates:

• Speed of filing products has been increased. Filed 5 new DMFs – 3 in US and 2 in EU
• Currently ramping up the facility in VIZAG with strong customer attraction, and will be commercialized as planned in Q3
• VIZAG EBIDTA growth guidance changed to 40%+ YoY

• Growth Driver: Broad based growth in other products and Vizag.

• PLI: Company has filed Mysore for PLI scheme for 1 molecule.

• Technology: Company is about to complete its 2 out of 3 technology, which will enhance efficiency for Solara.

Demand in some of sectors is moderated:
• Mgmt expected moderation in demand, hence company has taken necessary steps for those products.

Gross Margins:
• It was down a bit due to product mix however.

Fiscal 2022:
• Solara has grown in regulated market space.
• Vizag will kick in. CRAMS will support tailwind.
• Most of the CRAMS is commercial API.

• Most of the product is expected to be stable both in term of demand and price in the coming few quarter
SCIENCE
James Hay
James Hay
@jameshay218
Ct values can be used to estimate epidemic dynamics UPDATE! Ct values are expected to change depending on whether the epidemic is growing or declining, and we harness this to estimate the epidemic trajectory. Lots of cool new analyses and methods!

Highlights:
- Cts from symptom-based surveillance change over time, but the effect is weaker
- Methods to infer incidence using single cross-sections of Cts
- Unbiased by changing testing coverage
- Gaussian process (wiggly line) model for incidence tracking using Ct values
2/12

This work is *JOINTLY led* with @LeekShaffer and PI’d by @michaelmina_lab. Thank you also to the ever insightful @mlipsitch and to coauthors @SanjatKanjilal @gabriel_stacey and @nialljlennon. 3/12

Premise: times since infection depend on the epidemic trajectory. Distributions of randomly sampled viral loads proxy times since infection. With calibration, Ct values can estimate growth rate. We focus on qPCR in SARS-CoV-2, but the principle applies to any outbreak. 4/12

Result 1: viral loads are shifted higher (Cts lower) during epidemic growth and lower (Cts higher) during decline when individuals are sampled *based on the onset of symptoms*. We simulated linelist data under symptom-based surveillance and looked at TSI and Cts over time. 5/12
SCIENCE
DR22 \u03a9 \U0001f3ad\U0001f525
DR22 Ω 🎭🔥...
@DejaRu22
Best visual demonstration of God’s source code in action
SCIENCE

You May Also Like

Jonathan Ware
Jonathan Ware
@ReassessHistory
Rhino Barges

One of the dullest, coolest, more bizarre and fascinating pieces of kit used in Normandy.

Which NO ONE REALLY CARES ABOUT.

BUT I DO AND YOU SHOULD TOO.
/1

#WW2 #SWW #History


Planning for Overlord and Neptune had a serious snag, how to get troops from LSTs onto the beach as simply ramming them onto the beaches and dropping the ramp was known to damage the exceptionally vulnerable LSTs and felt to be unsustainable in the mid to long term. /2


LSTs were essential in sustaining Overlord's progress and were a subject of major headaches in the planning phase, and a real subject of friction when it came to launching additional amphibious operations such as Dragoon.

A single LSTs loss represented a capability nick. /3


So, logic dictated, we don't really want these remarkable but rare and highly valuable vessels constantly doing beach landings, we want something to act as a medium between ship to shore, and in came the (mostly) American designed Rhino. /4


I fucking love Rhinos.

Seriously.

Graceful lines of pure industrial pedigree.

Modular.

LOOK AT IT. /5
HISTORY
Anshul Pandey
Anshul Pandey
@Anshulspiritual
LING PURANA: IMPORTANCE AND CHARACTERISTICS

Reading Ling Purana provides bhog and moksha to the reader and listeners. It is about Shiva's sermon to Brahma when Shiva entered the Agni Ling and preached about Dharma sidhhi through the stories of Agni Kalp.


Vyasji divided this into two parts. This book is different from others and has eleven thousand shlokas which depict the glory of Shiva. This Puran is the summary about importance of Tridev.
It starts first with a questions. Then it describes the Shrishti in brief.

Then there is description of Yog and Kalp. Next is the evolution of Ling and its puja Vidhi. It consists of dialogue between Sanat Kumar and Shail etc. It also mentions the character of Dadhichi, the dharma representation of that yug, Bhuvan kosh description,

Chandravansh and Surya vansh. Next there is elaborate description of Shrishti, story of Tripur, Ling and Pashu Paash vimoksha, Vrats for Shiva, Discipline, Repentance, Arisht, description of Shri Shail. Next is the story of Andhakasur, Varah, Narsingh,

k¡lling of Jalandhar, Shiv sahastra naam, story of Daksha's yagya and its destruction, killing of Madan and marriage of Parvati.

Next is Vinayak katha, Shiv tandav, Ambrish katha and Upmanyu .
RELIGION
Advertisement
Kieran Snyder
Kieran Snyder
@KieranSnyder
1 There's a chasm between an NLP technology that works well in the research lab and something that works for applications that real people use. This was eye-opening when I started my career, and every time I talk to an NLP engineer at @textio, it continues to strike me even now.

2 Research conditions are theoretical and/or idealized. A huge problem for so-called NLP or AI startups with highly credentialed academic founders is that they bring limited knowledge of what it takes to build real products outside the lab.

3 A product is ultimately a thing that people pay for - not just cool technology or user experience. But I’m not even talking about knowledge gaps in go-to-market work. I'm talking purely technical gaps: how you go from science project to performant + delightful user experience.

4 Most commoditized NLP packages solve well-understood problems in standard ways that sacrifice either precision or performance. In a research lab, this is not usually a hard trade-off; in general, no one is using what you make, so performance is less important than precision.

5 In software, when you’re making something for real people to use, these tradeoffs are a big deal. Especially if you’re asking those people to pay for what you’ve made (can’t get away from that pesky GTM thinking). They expect quality, which includes precision AND performance.
MACHINE LEARNING
Anu Satheesh \U0001f1ee\U0001f1f3
Anu Satheesh 🇮🇳...
@AnuSatheesh5
Nanganallur Anjaneya Temple
#Chennai
#Tamilnadutemples 🚩

Bhaktha Anjaneya temple at Nanganallur in Chennai is one among tallest Anjaneyar Vigraham. Here Anjaneyar blesses with relief to all illness. Vigraham is about 32 feet height made of single  stone.
Anjaneyar here is 👇


Sri Vishwaroopa Aadhivyathihara Baktha Anjaneya Swamy.

Sthala Puranam says that, this is the place where Indra Deva hit Hanuman on the cheek by his Vajrastram. Sri Raghavendra Swami and Sri Kanchi Paramacharyar were behind in building this temple. Temple is constructed to


Realize power of Anjaneyar who is seen both in Ramayana and Mahabharatha.
Vadamalai is important offering to Anjaneyar here. It is believed that vigraham contain powers to heal any disease. Shri Ramanavami is celebrated here.

Jai Sri Ram
Jai Hanuman
🙏🕉🙏
ALL
Queen Preshiii
Queen Preshiii
@preshdeyforyou
Men actually cum faster when having sex with someone they love

Haha y'all fw this tweet very well 😀

So why we're here lemme show you guys WHAT IMPRESSES A MAN MOST IN BED

Sweetie I wanna remind you that every Good sex😍 is based on mutual respect and love❤, so make sure that the guy you’re fucking is worthy of you.

. and never waste effort on a guy who isn't worth it. But as long as you’re convinced of the hero-status of this guy, your endeavours will never be wasted❤.

So Below Are Things He's Secretly Dying for You to Try In Bed👇

• BABE FUCK HIM WITH THE LIGHTS ON😋💡

Men loves it when they see their dick going in and out of our pussy🍑 —They love it so much.

They love to see our creamy💦 pussy wet their dick🍆— They fucking love it 😍

You may be worried about what your husband thinks of your post-breast-feeding boobs, your C-section scar, or that at some point, he'll stop being attracted to you because you look a little — OK, a lot—different than you did on your wedding day.
LIFE