Diane H. Norback

GROWTH AND CHARACTERIZATION OF NORMAL HUMAN UROTHELIUM IN VITRO

SummaryA method for initiating rapidly growing cultures of normal human transitional cells from ureter and embryonic bladder specimens has been developed and quantified. A new microdissection technique was used to nonenzymatically separate the urothelium. The use of enriched medium containing 10 μg/ml insulin, 5 μg/ml transferrin, and 1 μg/ml hydrocortisone resulted in improved growth. The use of thin collagen gel substrates (0.6 ml/60 mm petri dish) resulted in 97% attachment of explants compared to 77% attachment on plastic. Explants grown on thicker collagen (2 ml/60 mm petri dish) showed, in addition to better attachment, enhanced growth of cells as determined both by measurements of colony size and cell density. Cultures of transitional cells that were initiated using explants could be passed three to five times using 0.1% EDTA for dispersion. Autoradiography of [3H]thymidine-labeled cells showed an initial phase of rapid cell division in primary explant cultures and restimulation of cell division in passaged cultures. Transmission electron microscopy showed that the cells growing out from the explants were continuous with the stratified urothelium maintained in the original explant. Stratification of transitional cells occurred in cultures of both ureter and embryonic bladder cells. Surface cells were joined near their apices by junctional complexes. Desmosomes and Golgi vesicles were present in all cells. Passage in culture did not alter the morphological characteristics of cells.

Chlorinated triphenyl-induced extensions of the hepatic endoplasmic reticulum.

Summary Ingestion of a highly chlorinated triphenyl by rats induces liver hypertrophy with proliferation of the vesicular smooth endoplasmic reticulum and formation of large concentric membrane arrays within the cytoplasm of the hepatocytes. At 21 days these changes result in increased hepatic function. Associated with the intracellular membrane alterations are increased phospholipid/protein, decreased cholesterol/protein, and altered specific enzyme activities of the microsomes. It is proposed that the phospholipid and cholesterol levels of the new membranes are compatible with an altered structure for localization of the foreign lipid soluble compound. The authors gratefully acknowledge the technical assistance of Miss Lois Abrahamson.

Argyrophilic nucleolar organiser region (AgNOR) staining in normal bone marrow cells.

Fifteen normal bone marrow aspirates were stained with the agyrophilic nucleolar organiser region (AgNOR) method. The results of the specific staining AgNORs as well as nuclear and cytoplasmic staining were analysed. A system was devised to characterise precisely the AgNORs present in the nuclei of bone marrow cells. Particular types of bone marrow cells had a characteristic AgNOR and non-AgNOR staining pattern. The bone marrow cells were identified easily and reliably with AgNOR staining and the method was especially useful for lymphocytes, plasma cells, erythroid cells, basophils/mast cells, monocytes and cells containing haemosiderin. The immature haemopoietic cells exhibited more and larger AgNORs than the more mature cells. It is concluded that AgNOR staining can be used to study bone marrow cells by providing additional information when used in conjunction with conventional stains.

Infectious mononucleosis progressing to fatal malignant lymphoma: a case report and review of the literature.

Epstein–Barr virus (EBV) infection is nearly ubiquitous. While most primary EBV infections are mild and self-limited, occasional patients will develop severe complications of infectious mononucleosis (IM). Potentially fatal complications include fulminant hepatic failure, virus associated hemophagocytic syndrome and lymphoproliferative disorders. We report the case of an apparently immunocompetent 21-year-old woman with IM that progressed within weeks from a polyclonal B cell proliferation to a fatal B cell lymphoma.

T helper-cell leukemia/lymphoma: Presentation as an acute immune-mediated illness

A 36-year-old man presented with an acute immunemediated illness characterized by leukocytoclastic vasculitis and polyarthritis. Evaluation of the synovial fluid, bone marrow, and peripheral blood revealed large numbers of abnormal lymphoid cells labeling a 4B4-positive, CD4-positive, IL-2 receptor-negative, helper T cells. Hypergammaglobulinemia, immune complexes, high levels of serum IL-2 receptors, serum antibodies against foreign alloantigens, and specific cytolysis of the patients leukemic cells by his normal CD8+ T lymphocytes suggest an interaction of the malignant cells and his normal immune cells. Thus, some of the rheumatologic symptoms leading to the diagnosis of leukemia appear to reflect an immunoregulatory imbalance manifested by B-cell hyperactivity, likely induced by the malignant helper T cells, and attempted regulation of his malignant T cells by normal lymphocytes.

The effect of diphenylhydantoin and cortisol on the cell cycle.

Abstract Normal human lymphocytes cultured in the presence of phytohemagglutinin were blocked in G0G1 when diphenylhydantoin (DPH) or cortisol 3.6 × 10-4 M was added at the beginning of culture. The suppression of culture growth was analyzed by flow cytometry and confirmed by [3H]thymidine incorporation and mitotic rate analysis. The correlation of these measurements with flow cytometry was good for DNA synthesis and excellent for mitosis. There was an additive effect on the G0G1 retention of cells when both drugs were present in the culture. These data may partially explain the suppression of cell-mediated immunity which occurs in DPH-treated patients.

CYCLOSPORINE A IN ADULT ONSET NEUTROPENIA

first patient, the tolerance of FIBI/HDM associated with the shortness of the post-graft aplasia, which lowers the risk of infection and is characteristic of PBSC autograft. might achieve satisfactory long-term remission in patients 50 years old. Thus, such a programme could be a new alternative for the treatment of patients with high risk multiple myeloma and justifies further studies. have been insufficient to allow a satisfactory engraftment. we infused him both PBSC and marrow prodiicts (1 x 104/kg b.w.) harvested when he was in remission (defined by the disappearance of monoclonal component in blood and by a normal marrow aspiration). Patient 2 received only the total quantity (30 x 104/kg b.w.) of PBSC collected. The extrahaematological tolerance was good. In both patients the duration of post-graft neutropenia was very short and they were discharged from LAF room from days 10 and 9. But if the platelet count rose rapidly in patient 2 ( > 50 x 109/1 on day 14, > 100 x 109/1 on day 27) and remains stable until now, it remained low in patient 1, needing maintenance platelet transfusions during 3 months, not exceeding now 3 5 x 10y/l. Furthermore, he presented an acute but transitory immune neutropenia on the third month post-graft. In spite of a persisting mild neutropenia (mean 1 x 109/1) in this patient, both he and the second one remain in complete remission 7 and 2 months after autograft. 11 and 6 months after diagnosis. respectively. In both patients, HDM has been well tolerated and has initially achieved a significant mass tumouir reduction. But the following collection of PBSC has been in unequal quantities, allowing only in patient 2 a pure PBSC autograft: the haemopoietic repopulating capacity of PBSC is now well established if the stem cell dose infused is 2 1 5 x 104/kg b.w. (Reiffers et al. 1987). Evidently, this new approach in the treatment of MM might be the most satisfactory in case of pure PBSC autograft, supposed to be uncontaminated with tumour plasma cells or precursors. But, even in the case of ‘mixed’ autograft (both PBSC and bone marrow) as in our