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Myelin was isolated from central white matter of beef and human brains. I t contained less than 8.1% ribonucleic acid, l e a than 8.63% deoxyribonucleic acid, and w s g l p l i p i d N-acetylneunminic acid. The preparation was 96% soluble in chioroform-methanol. Most of the insoluble materrai was sucrose. The myelin contained 19.6Y0 protein, 13.4y0 cholesterol, 86.1y0 phospholipid, and 23.6% galactslipid. A protein extracted from the rnyelin reparation with cold 0.2 N sulfuric acid gave a single sedimenting boundary in g e Spinco model E analytical ultracentrifuge. In 8 M ura-formate starch gels (pH 8.4) it migrated as a single band toward the cathode. Amino acid analysis revealed that the protein contained more than Z8Y0 dibasic amino acids, but little methisnine and no cystine. I t was rapidly hydrolyzed by trypsin. The molecular weight was estimated at 10,8W, When injected intradermally into guinea pigs, in suspension in Freund" adjuvant, it caused weight lms, a flaccid paralysis, occasional convulsions, and death, n pattern characteristic of experimental allergic encephalomyelitis. Evidence is cited for the presence of a t least one other protein in purified rnyelin.

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This report describes immunocytochemical and ultrastructural methods which led to the identification of Reovirus type 3 (Reo-3) in the porta hepatis of a patient with extrahepatic biliary atresia. The study indicates that Reo-3 antigenic sites are demonstrable by the avidin-biotinylated complex peroxidase method following formalin fixation and paraffin embedding, but are destroyed by freezing and thawing prior to fixation. Deparaffinization of the block and subsequent rembedding in epon-araldite did not alter immunoperoxidase staining. This procedure offered the advantage of higher light microscopic resolution of semithin (1 micron) sections and assisted in the selection of specific areas for ultrastructural studies. Localization of Reo-3 in extrahepatic biliary atresia was confined to a biliary remnant in which there were acutely inflammed, partially necrotic microscopic ducts. Electron microscopic examination of the immunoreactive sites revealed virus-like particles similar in appearance to Reo-3 particles in infected tissue culture cells. The observations presented here support previously reported serologic data which have shown an association between Reo-3 infection and extrahepatic biliary atresia.

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