I have been *waiting* for this type of extensive autopsy study to be be performed since I first learned about #COVID19. 3/2020-3/2021
N=44 autopsies; Brains = 11
Great care taken to be performed within 24 hrs of death.
None vaccinated. Prior to variants.
Preserved more than 10k specimens for downstream analysis across 85 distinct anatomic sites and body fluids.
Fresh tissue and body fluids were preserved for downstream analysis in RNA later by droplet digital PCR assay for SARSCOV2 RNA
5/ Assessed for qPCR for subgenomic RNA which is thought to be a marker of recent viral replication.
As well as high throughput single genome amplification and sequencing to compare diversity of #SARSCOV2 spike gene within and across anatomic compartments.
6/ Flash frozen tissue and fluids were collected for cell culture to evaluate for replication competent virus.
Research tissues were formalin fixed for histopathologic analysis as well as in situ hybridization (ISH) and immunohistochemistry (IHC) to query cellular tropism.
7/ Defined persistence as detection of all RNA in tissues > or = to 31 days post initial #COVID19 symptom onset. Detected subgenomic RNA in > 300 samples.
Then tested 50 samples from 20 patients in vero culture and successfully cultured virus from 25 anatomic locations in 9 pts
8/ In these tissues, a cytopathic effect and an increase in #SARSCOV2 subgenomic RNA levels in cell culture supernatant relative to levels in original tissue homogenant were observed confirming replication competent virus.
9/ 100% of tissues with a subgenomic RNA cycle threshold between 15 and 20 had replication competent virus.
This number decreased to 64.3% of tissues that had CT value between 20 and 25.
And further decreased to 31.1% of tissues that had a CT value between 25 and 30.
10/ No replication of virus was detected in tissues with cycle threshold of 30 or higher.
Isolated replicating #SARSCOV2 from nasal placode, sinus turbinate, bronchus, lung, cornea & sclera of the EYE!, the jejunum, the mediastinal lymph node, the heart, and the adrenal gland.
11/ Robust framework showing that you can perform RNA and/or subgenomic RNA on tissue to determine if it likely to have replication competent virus without having to perform cell culture in a BSL3 setting (a limited resource)
12/ Using the ddPCR and sqRNA qPCR as a guide, we used ISH probing the SPIKE region of the SARSCOV2 genome to validate our PCR assays & to determine cell type specificity within tissues
Positive staining found in all tissue groups, small panel here that highlights ISH staining:
13/ Displayed in first row, in order from left to right
a) Frontal Lobe
b) Corpus callosum (early case)
c) Cerebellum (late case)
d) Cervical spinal cord (late case)
14/ Second row:
e) Follicular cells of Thyroid
f) Stratified squamous epithelium of the esophagus + stroma of a capillary (#)
g) Mononuclear leukocytes within the white pulp of the spleen
h) Colonic epithelium (*) and mononuclear cells within the stroma of the appendix (#)
15/ Third row:
i) Endocrine secretory cells of the adrenal gland
j) Stromal cells of the post menopausal ovary
k) Sertoli cells and maturing germ cells of the seminiferous tubule of the testes
l) Endometrial gland epithelium and stromal cells of a PREMENOPAUSAL UTERUS
16/ Evaluated SARSCOV2 cellular tropism within the cerebellum & other regions of the brain via fluorescent IHC and confocal microscopy.
Blue = cerebellum cell nuclei
Green = #SARSCOV2 nucleocapsid
Neurons = magenta
ML = molecular layer
GL = granular layer
WM = white matter
17/ A) Low magnification visualization of no antibody control
B) All specific protein expression (higher magnification of B shown in C, D, E shows specific cell type infection)
18/ C) Positive neurons indicated by yellow arrows, other unidentified cells indicated by white arrows are also associated with viral protein in the granular layer
D) Purkinje cells adjacent to the molecular layer are infected
19/ E) In rare instances, blood vessels adjacent to the granular layer and white matter were associated with viral protein
20/ This table summarizes the 36 cell types and hyaline membranes across the body and brain that were ISH positive for #SARSCOV2 SPIKE RNA in our cohort
21/ Final study aim was to determine if #SARSCOV2 evolves in different anatomical compartments within the same patient.
High throughput single genome sequencing to analyze SPIKE gene variant sequences from 46 tissues from 6 patients.
P18 = identical across all compartments
22/ P19 = Detected a single synonymous mutation (blue) in a minor variant in the right superior lung lobe
Overall, these 2 patients demonstrate that #SARSCOV2 can infect cells in non-respiratory tissues without requiring changes in the SPIKE genome.
23/ P27 = synonymous mutations (blue) were detected both within the respiratory tract and outside of it, specifically in the mediastinal lymph node and the left and right ventricle of the heart.
24/ P33 = Unable to sequence many tissues as a late case, there were 2 distinct haplotypes detected within the thoracic lymph node
25/ They sequenced brain samples from 2 patients, which revealed non-synonymous mutations between haplotypes within the CNS and the rest of the body, specifically in the dura mater for P36 and the hypothalamus and thalamus in P38. #COVID19#NeuroCOVID
26/ "These results indicate within the brain, that #SARSCOV2 is subject to the development of genetic compartmentalization...and exploring the mechanisms behind this will be...important... future...work..." (trails off somberly) #COVID19#NeuroCOVID#LongCOVID
Most dermatologists do not recommend using Neosporin d/t risk of allergic contact dermatitis. Declared the Contact Allergen of the Year for 2010 by the American Contact Dermatitis Society (ACDS). Can cause secondary dermatosis known as autosensitization. ncbi.nlm.nih.gov/pmc/articles/P…
Derms recommend polysporin instead because it doesn’t contain neomycin, just polymixin & bacitracin. Would also be curious what infectious disease physicians think of prolonged use of topical antibiotics in an era of increasing antimicrobial resistance & antibiotic stewardship…
This is particularly a concern for people who have altered immune systems, autoimmunity or issues with allergies already like with MCAS.
The biology of neurotypical (allistic) brains is towards maximal efficiency. Microglia literally prune redundant synapses to strengthen neural connections. This works differently in autistic folks who have many more dendritic spines & less pruning🧵
Paper: nature.com/articles/nn.27…
2/ These brains function differently. Many folks have superior pattern recognition. It’s not fully understood why. Some areas of the brain have shown more connectivity, some less. Many variables at play and lack of diagnosis especially in underrepresented groups are significant.
3/ Some literature cites higher incidence of neurodegenerative diseases in Autistic people but with the known diagnostic and representation issues, as well as how much we don’t understand about the brain, I’m not sure how much of that can be extrapolated to an entire population.
This is a really interesting thread on eugenics but has some glaring omissions like the connection with the current pandemic, ongoing US taxpayer funded genocide of Palestine, or that our country was founded on the genocide of Indigenous Americans and built by enslaved Africans🧵
2/ The glaring foundation of white supremacy, colonialism, patriarchy, ableism and capitalism should also be emphasized. Nazi propaganda of “useless eaters” is particularly relevant to our current moment in justifying the mass death of disabled people (as we disable more people)
3/ One could practically do a dissertation on the pandemic specifically explaining the harmful rhetoric and messaging from our administration, Walensky & Fauci regarding eugenics of “the vulnerable” when it’s *society* that MAKES people vulnerable
It has *ALWAYS* been a workers rights issue. One could argue one of the largest in history.
If the people in power won’t protect themselves or their own families from infection, how little do you think they care about you and me? They prioritized “the economy” above all else.
Computer based tests like this exist and patients can do them remotely at home. Because many patients with LC are severely disabled and just getting to a clinic is cognitively and physically exhausting.
The discovery of Hep B Virus was an EMBARASSMENT to @theNCI exactly because of the heavily funded and largely failed *Special Virus Cancer Program* from a scientist who LEFT the NCI in the 1960s exactly because of his interdisciplinary curiosity! Work that led to the HBV vaccine.
@theNCI The first virus discovered to cause cancer in humans was EBV (virus that causes mono) and Burkitt’s Lymphoma in 1958 by an Irish surgeon, Dennis Burkitt and two British virologists.
Caused a whole scurry of chaos with media printing “Cancer may be infectious!” in @LIFEmagazine.