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Biomedical subjects

R S Basch

Publications and source records attributed to R S Basch.

65 records · Page 4Linked to original sources

3 -Hydroxy steroid dehydrogenase activity in the mitochondria of rat adrenal homogenates.

The activity of 3beta-hydroxy steroid dehydrogenase (EC 1.1.1.51) in the mitochondrial fraction of rat adrenal homogenates was approx. 31% of the total activity recovered after differential centrifugation and washing of the particulate fractions. Some 45% of the total activity was found in the microsomal fraction. The activity was assayed by a radioisotopic method devised in this laboratory for the purpose of studying small quantities of tissue and cell fractions. Satisfactory separation of the two fractions was demonstrated by electron microscopy of the pellets and by comparative recoveries of RNA, steroid 21-hydroxylase and cytochrome c oxidase in the various compartments. Analyses of the kinetics of the enzyme activity in the two fractions revealed no significant differences in apparent K(m) for pregnenolone, dehydroepiandrosterone or NAD(+), but demonstrated a distinct difference in the K(m) for NADP(+). pH optima and susceptibility to cyanoketone inhibition were similar in both fractions.

Adrenal Glands↗

Early thymic regeneration after irradiation.

Whole body irradiation produces profound thymic atrophy. After sublethal irradiation, regeneration begins promptly and the earliest regeneration is from radioresistant intrathymic precursors. The progeny of these precursors expand rapidly and restore thymic cellularity to near normal within 2 weeks. We have used monoclonal antibodies specific for a variety of differentiation markers of the T lineage to analyze the early events in thymic regeneration. A three-color flow microfluorometric analysis revealed that the majority of the cells found early in the regenerative process have the phenotype of mature T cells. These include CD4-/CD8-; CD3hi as well as CD4+/CD8-; CD3hi and Cd4-/CD8+; CD3hi. The proportion of cells with mature phenotypes declines rapidly between day 6 and day 12. Not all of the early appearing cells have mature phenotypes. Among the early cells that do not express CD3 are both CD4 and CD8 single positive cells that express HSA and resemble the intrathymic precursors found in other systems. In these mice CD4 single positive predominate. There are other cells that are HSA positive but express low levels of CD4 and very low levels of Thy-1. These appear to include the earliest members of the T-lineage. In addition to relatively mature conventional T cells and early progenitors, the early developing population includes cells that express markers of the T-cell lineage including the T-cell receptor but do not express Thy-1. These Thy-1 negative T cells comprise a significant number of the earliest cells found after regeneration.

Animals↗

Isolation of cell lines possessing functional and serological properties resembling those of thymocyte precursors.

Thymocytes develop from a committed haematopoietic progenitor, referred to as a prothymocyte. They are uniquely capable of migrating to and restoring the thymus of a lethally irradiated host, a property which has been exploited as a specific assay for these cells. Like other committed haematopoietic progenitors, prothymocytes are found only in small numbers in even the richest sources (0.05-1.0% of the nucleated cells in bone marrow). Purification has proved difficult both in terms of finding a suitable starting material and in the degree of enrichment achieved. We now report the isolation of cloned lines of cells with some of the serological and functional properties of prothymocytes. One of these lines has been in continuous culture for almost 2 years. When injected into irradiated recipients, cells from this line migrate to the thymus and there develop into cells which resemble normal cortical thymocytes.

Animals↗