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Abstract— Sensitive micromethods were used to study the plaques, adjacent white matter and remote, grossly normal white matter from two cases of multiple sclerosis and to compare them with white matter from normal controls. Lipid‐free dry wt/unit of volume was found to be similar for plaques and for normal white matter, reflecting a high water content of plaque tissue and establishing a base for comparison of enzyme activities. Elevations of acid proteinase in and around plaques were confirmed, but they were far exceeded by the increases in acid phosphatase; other acid hydrolases (β‐galactosidase, β‐glucuronidase and dipeptidyl arylamidase II) showed no significant or consistent changes. However, an acid lipase‐esterase hydrolysing 4‐methylumbelliferyl oleate was about 30% as active in plaques as in normal‐appearing white matter. Glucose‐6‐phosphate dehydrogenase was unchanged except in one plaque, but lactic dehydrogenase was markedly elevated both in plaques and adjacent white matter. The grossly normal white matter of MS patients, although histologically far from normal (showing gliosis, perivascular infiltrations and small plaques), did not differ significantly from controls with regard to the activity of any of the enzymes studied. DNA levels were much reduced in plaques, but comparisons were difficult because of the apparent gliosis in normal white matter. Decreases in dry wt/unit vol, reflecting partial demyelination, could be shown to extend in a gradient to a distance of about 2 mm. from the edge of certain plaques.

—In Lewis rats with experimental allergic encephalomyelitis (EAE), quantitative histochemical techniques were used to measure the activities of several hydrolytic enzymes, especially acid proteinases, and their levels were correlated with the concentrations of DNA and total lipids. Only those samples which contained lesions (visualized directly or in stained nearby sections) had elevated acid hydrolase activities. A lipase‐esterase hydrolysing a fluorogenic substrate at pH 5.0 was more active in lesions; activity on the same substrate at pH 7.2 was not elevated. β‐Glucuronidase rather than acid proteinase or cathepsin D was the enzyme showing the greatest differences. In a monkey with EAE, studied for comparison, a massive inflammatory lesion appeared to leak β‐glucuronidase to a distance of about 2.5 mm beyond its margin.

Abstract— A thiol dependent proteolytic enzyme (tentatively identified as carboxypeptidase B) which liberates phenylalanine from CBZ‐glutamyl‐phenylalanine at pH 5.3 was shown, by a sensitive micromethod, to be greatly increased in activity in and around MS plaques. These increases exceeded those of other hydrolases previously measured. Plaque tissue, on the basis of lipid‐free dry weight, is up to 3‐fold richer in this enzyme than control white matter, and most samples of apparently uninvolved MS white matter already show elevated activities. The enzyme is highly dependent on the presence of dithiothreitol. It is unaffected by diisopropyl fluorophosphate and pepstatin, but inhibited by iodoacetate and leupeptin. Macrophages or lymphocytic infiltrations in the tissue do not appear to be the main source of the enzyme. In conjunction with measurements of DNA, reflecting gliosis, invasion by hematogenom cells and proliferation of phagocytes as well as oligodendrocyte loss, and acid lipase‐esterase, indicative of the survival or degeneration of oligodendrocytes, these results are interpreted as probably reflecting predominantly the activity of astrocytes. The incidental finding that most samples of unaffected white matter from MS patients contain more DNA per unit lipid free dry weight than average control white matter is considered significant in pointing to more widespread tissue changes independent of or preceding plaque formation.

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Abstract— Arylsulphatase activity, with 4‐methylumbelliferone sulphate as substrate, was measured by a quantitative histochemical method in individual anterior horn nerve cell bodies and adjacent neuropil of man and monkey; and in molecular and granular layers and subjacent white matter of cerebellum of monkey, rat and guinea pig. The activity was much higher in neuronal perikarya than in neuropil, and higher in the granular layer of cerebellum than in the molecular or white matter, thus resembling the distinctive distribution, reported in monkey, of three other lysosomal enzymes, β‐galactosidase, β‐glucuronidase and α‐naphthyl acid phosphatase. One exception was encountered: the white matter of guinea pig cerebellum had more arysulphatase activity than the granular layer. For comparison, other lysosomal enzymes also were measured in rat and guinea pig cerebellum; in these species, α‐naphthyl acid phosphatase distribution was found to differ from that of β‐galactosidase and arysulphatase, and from the pattern common to four lysosomal enzymes in the monkey.

Abstract— The localization of an acid lipase‐esterase which cleaves the fluorogenic substrate 4‐methyl‐umbelliferyl oleate at pH 5 was studied, because previous experiments has shown this activity to be reduced in the plaques of multiple sclerosis. In the human cerebellum, quantitative histochemical methods showed the activity to be relatively low in the molecular layer, compared to the granule cell layer; but the underlying white matter was the most active. In the human spinal cord, anterior horn cell bodies were richest in acid lipase, but white matter was, on a dry weight basis, as active as neuropil. Oligodendrocytes obtained in bulk from bovine white matter according to Poduslo & Norton (1972) had a specific activity up to 20 times greater than the crude myelin fraction. While it remains possible that in myelin the enzyme is in an inactive or inhibited state, the results indicate that the enzyme is localized in oligodendroglial cell bodies and suggest its use as a marker.