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Autopsy in Myalgic Encephalomyelitis
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==== Chia 2015 study ==== A | 2015 study<ref name=":0">{{Cite web | url = https://forums.phoenixrising.me/index.php?threads/are-infections-just-a-trigger-of-me-cfs-or-an-ongoing-cause-of-me-cfs.37549/page-25#post-632251 | title = Chronic enterovirus infection in a patient with myalgic encephalomyelitis / chronic fatigue syndrome (ME/CFS) β clinical, virologic and pathological analysis. John Chia, David Wang, Andrew Chia, Rabiha El-Habbal. 2015. Presented at the 19th International Picornavirus Meeting, 2016. | date = |access-date= | website = | last = | first = |archive-url=|archive-date=|url-status=}}</ref> by [[John Chia]] looked at the brain tissues of another ME/CFS patient, a 23 year old male with high titers to [[echovirus]] 11 who who committed suicide 6 years after the onset of symptoms. Using Western blot, Chia found evidence of enterovirus infection in the pontomedullary junction and midbrain (both are in the brainstem), medial temporal lobe, lateral frontal cortex, occipital lobe and cerebellum. And by RT-PCR, Chia found enterovirus RNA in the frontal cortex, pontomedullary junction. When the virus was sequenced, they found a 92% homology to CVB2, and about 86% to echovirus 30.<ref>{{Cite web | url = http://www.investinme.org/IIMEC10.shtml | title = Dr John Chia's presentation at the Invest in ME London Conference 2015 (available on DVD), timecode 27:32. | last = | first = | date = | website = | archive-url = | archive-date = |url-status = | access-date=}}</ref> Interestingly enough, the first time Chia used RT-PCR to find enterovirus RNA in this patient's brain tissue, he got a negative result. But Chia remembered a phenomenon whereby viral RNA may bind to chromosomal [[DNA]], preventing RT-PCR from detecting the RNA. So Chia used the DNase [[enzyme]] to digest the chromosomal DNA in the brain tissue, and after doing this, he was able to find enterovirus RNA in the frontal cortex and pontomedullary junction.<ref>{{Cite web | url = http://www.investinme.org/IIMEC10.shtml | title = Dr John Chia's presentation at the Invest in ME London Conference 2015 (available on DVD), timecode 26:46 | date = |access-date= | website = | last = | first = | archive-url = | archive-date = |url-status = | quote=So when we did our RT-PCR again it's completely negative. That's the positive control here, negative control here. Then we remembered one phenomena: so we actually use the DNase to try to digest any chromosomal DNA off this RNA. We think they are attached like this. Then we repeated the RT-PCR: then we saw the band here, for the pontomedullaryjunction, and this one's the frontal cortex, here's the positive strand, the positive control and negative control. Then we sequenced it: this is our top band for the pontomedullary junction, OK, and that's clearly different from the positive control. And that was about 92% homology to CVB2, about 86% to echovirus 30.}}</ref> Dr Chia points out that an enterovirus infection in the stomach can travel along the [[vagus nerve]] (via reverse axonal transport) and reach the brainstem in around 3 days. In this way, the virus can bypass the [[blood-brain barrier]], and enter into the brainstem, infecting the brain.<ref>{{Cite web | url = https://www.youtube.com/watch?v=obHtCwhg3-0&t=2m20s | title = Dr John Chia's presentation at the State of Knowledge Workshop on ME/CFS, National Institutes of Health, April 2011, day 1, part 1, timecode 2:20. | date = |access-date= | website = | last = | first = |archive-url=|archive-date=|url-status=}}</ref> Chia also questions the common wisdom that in brain infection, you will find the virus in the cerebrospinal fluid: he points out this is not always true in the case of enterovirus, because even in acute enterovirus 71 infection, only 5% of the cerebrospinal fluid samples are positive, and ME/CFS does not involve an acute brain infection, but rather a chronic one. Dr Chia points out that in ME/CFS, enterovirus infections are found in the tissues, not really so much in the blood and fluids.<ref>{{Cite web | url = https://www.youtube.com/watch?v=BO-yxqZuXTY&t=9m34s | title = Dr John Chia's presentation at the State of Knowledge Workshop on ME/CFS, National Institutes of Health, April 2011, day 1, part 2, timecode 9:33. | last = | first = | date = | website = | archive-url = | archive-date = |url-status = | access-date=}}</ref> This is because the persistent enterovirus infections found in the tissues of ME/CFS patients are of the aberrant [[non-cytolytic enterovirus]] form. The following table summarizes the areas of the brains of ME/CFS patients in which enterovirus infection was found post-mortem: {| class="wikitable" cellpadding="0" !Brain Study !Date !Enterovirus Infected Areas in Red !Enterovirus Testing Method !Areas Of Brain Found Infected With Enterovirus |- style="background-color: white;" |McGarry, Gow and Behan <ref name="pmid8172448" /> |1994 |[[File:Brain Small McGarry Brain Autopsy.jpg|frameless|200x200px]] |PCR |Hypothalamus and brainstem. |- style="background-color: white;" |John Richardson <ref name="Richardson2011" /> |2001 |[[File:Brain Small Richardson Brain Autopsy.jpg|frameless|200x200px]] |VP1 protein stain |Fibroblasts in the adventitia of small blood vessels in the cerebral cortex, and also in a small fraction of the glial cells. |- style="background-color: white;" | rowspan="2" |John Chia <ref name=":0" /> | rowspan="2" |2015 | rowspan="2" |[[File:Brain Small Chia Brain Autopsy.jpg|frameless|200x200px]] |Western blot |Pontomedullary junction and midbrain (both are in the brainstem), medial temporal lobe, lateral frontal cortex, occipital lobe and cerebellum. |- style="background-color: white;" |RT-PCR |Frontal cortex, pontomedullary junction. |}
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