Barbara E. Barker

HUMAN MAMMARY SLICES IN ORGAN CULTURE. I. METHOD OF CULTURE AND PRELIMINARY OBSERVATIONS ON THE EFFECT OF INSULIN.

Abstract A technique for culture of human mammary slices has been devised, which permits survival and limited proliferation of all elements for periods up to 12 days in vitro. Slices survive in a variety of media with or without serum, providing that glucose concentrations are maintained above critical levels. In these organ cultures, addition of insulin to the medium results in marked morphological changes of ductal epithelium including proliferation, hypertrophy, and foci resembling squamous metaplasia.

HPLC analysis of aromatic amino acids, nucleosides, and bases in plasma of acute lymphocytic leukemics on chemotherapy

Plasma chromatograms--obtained by the reversed-phase mode of high performance liquid chromatography (HPLC)--of 19 subjects with acute lymphocytic leukemia (ALL) were compared to those of 19 normal individuals. ALL patients were in remission and on a methotrexate and 6-mercaptopurine maintenance regimen. The concentrations of the aromatic amino acids L-phenylalanine and L-tyrosine, the nucleosides uridine, adenosine, inosine, and guanosine, as well as the bases hypoxanthine and xanthine, were elevated in the leukemic in comparison to the normal chromatograms. Highest inosine levels corresponded to leukemic subjects whose condition severely deteriorated with time. Patients with lower inosine levels are still in continuous remission.

Cytochemical studies of human bone marrow fibroblast-like cells. I. Alkaline phosphatase.

Abstract Alkaline phosphatase activity of marrow fleck components and fibroblast-like cells (FLC) developing in marrow cultures has been compared. The enzyme is demonstrable in the vascular network of marrow stroma, maturing myeloid cells, and FLC developing in culture. The vascular network of marrow flecks visualized by this method is described. It is concluded that the FLC originate from marrow stroma and not from any hemic cells, in this culture system. The most likely parent cell of the FLC is the endothelial cell.

Phytomitogens and lymphocyte blastogenesis

I n the past 8 years the response of human blood lymphocytes to a variety of mitogenic substances has attracted increasing attention and many new investigators have entered the field of hemic cell culture. The modulation of the blood lymphocyte to a primitive blast-like cell, inappropriately termed transformation, has more recently been designated blastogenesis. This is the term which I will use in discussing our work. It will probably be helpful to trace briefly the development of this field in order to place our own interests and observations in the proper perspective. In 1949 Li and Osgood at the University of Oregon introduced phytohemagglutinin, an extract from the red kidney bean Phaseolus vulgaris, to hemic cell culture as a hemagglutinin for separation of erythrocytes from leukocytes (1). In 1960 Peter Nowells epoch-making demonstration that the phytohemagglutinin (PHA) was capable of stimulating a blast-like modulation of blood mononuclear cells (2) opened the door for development of practical techniques in human and animal cytogenetics. Nowell accurately characterized the morphologic features of the blastogenic process. When human white blood cells were placed in vitro in the presence of phytohemagglutinin some lymphocytes showed no response. Many, however, quickly became activated. They became much larger, assumed a primitive blastic appearance by 3 days in vitro with large nuclei and prominent nucleoli, and possessed deeply basophilic vacuolated cytoplasm. Much mitosis was

Effect of the methanol extraction residue of Bacillus Calmette-Guerin in advanced Hodgkin's disease

One‐hundred‐ninety‐six patients with Stage III and IV Hodgkins disease were prospectively randomized to receive either treatment with the methanol extraction residue of Bacillus Calmette‐Guerin (MER/BCG) or no immunotherapy. Prior to the MER/BCG randomization, patients received six courses of induction and two years of maintenance chemotherapy so that a group with a presumptively low tumor burden could be established. Only patients achieving a complete remission were evaluated. During the first two years of immunotherapy, the MER/BCG group had a relapse frequency twice that of controls. The overall crude relapse frequency and disease‐free survival were similar between the two treatment groups. The MER/BCG dose schedule used in this study was associated with a high frequency of unacceptable toxicity. Ulcerations of greater than 1 cm occurred in one‐third of the patients with associated pain, fever, and occasional lymphadenopathy. A high degree of patient noncompliance (36%) was observed. Age (P = 0.002), prior radiotherapy (P = 0.032), and chemotherapy (P = 0.044) were prognostic factors found to significantly influence remission duration. These factors were balanced between patients treated with immunotherapy and those who were not. MER/BCG therapy did not significantly delay or prevent relapse.

Tissue culture studies of human bone marrow: II. Protein synthesis in haemic cells

Abstract Human bone marrow cells cultivated in a chemically defined system incorporate tritiated histidine, phenylalanine, or leucine in vitro, as demonstrated by autoradiographic studies. Megakaryocytes show intense labelling up to 3 days, and platelet formation also occurs. Immature members of the granulocytic series, both neutrophilic and eosinophilic, show heavy labelling. Promyelocytes show greatest amino acid incorporation, and mature neutrophiles and eosinophiles show the least. Erythroblasts label in early cultures, and occasional erythrocytes, probably reticulocytes, also show significant incorporation. The presence of prednisolone in the medium is associated with increased incorporation of precursor as compared to that of cells cultured in hormone-free medium.

A technique for chromosome study of human bone marrow fibroblast-like cells☆

Abstract 1. 1. A technique for chromosome analysis of fibroblast-like cells from human bone marrow tissue cultures is presented. 2. 2. This technique may also be applied to the study of skin fibroblasts from skin explants.

Unusual blasts with basophilic granules in 2 cases of de novo acute myeloid leukemia with inv3(q21q26.2) and monosomy 7 and coexpression of CD2 and CD31.

Acute myeloid leukemia with inv3(q21q26.2)/t(3,3)(q21;q26.2) is a subtype of acute myeloid leukemia associated with significant dysmyelopoiesis and a poor prognosis. In more than a half of the cases, there is also monosomy 7. We present 2 young male patients with de novo acute myeloid leukemia with inversion 3 and monosomy 7 who had significant morphologic and immunophenotypical similarities. Both patients had circulating subsets of blasts with unusual intracytoplasmic basophilic granules and prominent bone marrow dysmegakaryopoiesis. The leukemic myeloid blasts were negative for myeloperoxidase and had aberrant coexpression of CD2 and CD31. Despite their morphologic and immunophenotypical similarities, only 1 of the patients achieved remission and remained free of disease 24 months after bone marrow transplant. The younger patient, who had also increased hemoglobin F and an associated FLT3 D835 variant, had an acute myeloid leukemia refractory to chemotherapy and died 4 months after his diagnosis.