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Biomedical subjects

G R Johnson

Publications and source records attributed to G R Johnson.

At least 199 records · Page 11Linked to original sources
PubMed1982

Differentiation and "renewal" of multipotential cells in vitro.

Cells obtained from in vitro colonies expressing multiple hemopoietic lineages (mixed-erythroid colonies) have been shown to produce secondary mixed-erythroid colonies. When individual mixed-erythroid colonies are studied, considerable variation has been observed in the capacity to produce secondary mixed-erythroid colonies. That this heterogeneity in self-renewal capacity may be an intrinsic property of the primary mixed-erythroid colonies has been shown by the ability to fractionate fetal liver mixed-erythroid colony-forming cells with high and low levels of secondary colony formation. In addition mixed-erythroid colony-forming cells obtained from spleens consistently produce higher numbers of secondary mixed-erythroid colonies (up to 250 secondary mixed-erythroid colonies per primary mixed-erythroid colony) than bone marrow-derived mixed-erythroid colonies (no secondary mixed-erythroid colonies obtained) suggesting that external factors may predetermine the ability of primary mixed-erythroid colony-forming cells to self-renew in vitro. The ability of supernatants obtained from long-term marrow cultures to enhance secondary colony formation by bone marrow-derived mixed-erythroid colony-forming cells raises the possibility of defining the nature of the proposed external factors that determine stem cell self-renewal.

Animals↗
PubMed1982

The production of committed hemopoietic colony-forming cells from multipotential precursor cells in vitro.

Murine fetal liver cells have been fractionated by fluorescence-activated cell sorting into two fractions termed. For convenience, the pre-CFC and CFC (colony-forming cell) fractions, which differ in their relative binding of the lectin, pokeweed mitogen. The CFC fraction contained a high frequency of CFC (26%) and in liquid cultures stimulated with colony-stimulating factors generated large numbers of differentiated progeny. Consequently, residual CFC could not be detected in such cultures after day 4. In contrast, the pre-CFC fraction contained relatively few CFC (1%) but contained the majority of CFU-S and was able to generate large numbers of new CFC after 5-7 days in liquid culture. This production of CFC from precursor cells was absolutely dependent on factors present in pokeweed-mitogen-stimulated spleen cell conditioned medium (PWM-SCM) or postendotoxin serum (ES) but was not stimulated by known granulocyte or macrophage colony-stimulating factors. CFC production occurred from multipotential precursor cells and all types of CFC were detected (excluding lymphocytes). Clonal analysis of pre-CFC showed that most of the generated CFC arose from relatively few multipotential precursors some of which could generate up to 500 CFC. The data suggested a differentiation sequence of the multipotential precursor cells in which differentiation potentials are successively restricted in the order macrophage, eosinophil, granulocyte-macrophage leads to granulocyte, megakaryocyte, erythroid. The frequency of CFC-generating cells in the pre-CFC fraction was significantly less (ten-fold) than that of CFU-S, suggesting that pre-CFC may be a more ancestral subset of CFU-S. The cell fractions and assay systems described should be of use in defining and purifying factors regulating early phases of hemopoietic differentiation and in defining the differentiation steps involved in the restriction of potentialities of multipotential cells.

Animals↗
PubMed1982

Colony-forming cell proliferation: a rapid and sensitive assay system for murine granulocyte and macrophage colony-stimulating factors.

A microculture assay for murine granulocyte-macrophage colony-stimulating factor (GM-CSF) has been developed using fetal liver GM colony-forming cells (CFC) isolated by fluorescence-activated cell sorting. These GM-CFC are free of mature hemopoietic cells, such as granulocytes and macrophages, which may interfere with direct assays for GM-CSF. The assay procedure allows the quantitation of GM-CSF within 48 hr by measuring the number of cells produced from 50 GM-CFC in microcultures (15 microliter). The assay is particularly simple to set up and score and yet, because of the reduced volumes, this assay is still capable of detecting 0.2 pg (i.e., 0.2 U) of GM-CSF within 48 hr, i.e., 100 times less GM-CSF than the conventional soft agar assay. By allowing the microcultures to develop for 7 days, the extra proliferation allows a further tenfold increase in the sensitivity of CSF detection. The time and cost of setting up hundreds of GM-CSF assays for fractions from chromatographic columns, e.g., reverse phase high performance liquid chromatography, is reduced by at least five-fold. Enough GM-CFC can be isolated and stored frozen in one afternoon to provide sufficient cells for the daily assay of 200 samples of GM-CSF for several months. Microassay results for several sources of GM-CSF at different stages of purification are compared to the results obtained from the soft agar assay.

Animals↗
PubMed1981

Cost effective moulded seating for the handicapped child.

For many severely handicapped children requiring external trunk support there is probably no better solution than to provide custom moulded seating. This will improve the patient's posture, reduce the occurrence of pressure sores and facilitate such activities as feeding. However, if large numbers of patients are to be supplied with this type of equipment, it is important that it should be manufactured at the lowest cost compatible with an acceptable quality. These manufacturing problems have been studied in considerable detail and a production method using vacuum consolidation casting together with a modified vacuum forming process has been developed. This has made it possible to produce custom moulded seating efficiently with the minimum of capital equipment. Much attention has been paid to the final appearance of the article and this has been enhanced by the use of commercially available items for the supporting structure and for the finishing of edges, etc. It is suggested that other orthoses could be redesigned for production using an approach similar to that described in this paper.

Child↗
PubMed1981

Presence of multipotential hemopoietic cells in teratocarcinoma cultures.

Previous investigations have shown that the teratocarcinoma line PCC3/A/1 is able to differentiate in vitro to produce red blood cells of the primitive hemopoietic cell population of the mouse embryo. The present paper demonstrates the presence in the same cultures of multipotential cells capable of giving rise to colonies containing erythrocytes, macrophages, and megakaryocytes when stimulated by pokeweed-mitogen spleen-cell conditioned medium.

Animals↗
PubMed1981

A serological survey of Leptospira interrogans serotype pomona in Alberta and Saskatchewan striped skunks and possible transmission between cattle and skunks.

The range of known occurrence of Leptospira interrogans serotype pomona is extended to Alberta in striped skunks (Mephitis mephitis); no evidence of L. sejroe was found. Reacting sera from skunks were confined to the southern portion of Alberta and adjacent Saskatchewan, although a number of reactors were found sufficiently further north in Saskatchewan suggesting that a different mode of infection may be functioning there. Of 95 skunk sera from near a dairy farm infected with serotype pomona 40% were reactors. Of 438 skunk sera from other areas only 5.7% were reactors; that difference was suggestive of transmission from cattle to skunks on the dairy farm. Of 22 skunk sera collected near the dairy farm in summer none were reactors, whereas 52% of skunk sera taken the previous and following winters were. That seasonal difference was not evident among sera from other locations.

Alberta↗
PubMed1980

Differential expression of lectin receptors during hemopoietic differentiation: enrichment for granulocyte-macrophage progenitor cells.

Molecular changes occur at the surface of hemopoietic cells during differentiation from progenitor cells to mature granulocytes and macrophages. The differential expression of surface carbohydrate residues has been probed using lectins and the results used to purify normal mouse granulocyte-macrophage progenitor cells. Ten different lectins were screened for selective interaction with mouse hemopoietic colony-forming cells (CFCs), using agglutination or a quantitative analysis of the number of fluoresceinated lectin molecules bound per cell using a fluorescence activated cell sorter (FACS). Pokeweed mitogen (PWM), Helix pomatia agglutinin (HPA), soybean agglutinin (SBA), and peanut agglutinin (PNA) preferentially bound to CFCs so that it was possible to enrich 4 to 10-fold for these progenitor cells by sorting for the highly fluorescent cells. Further analysis of the low and high angle light scattering characteristics of the CFCs indicated that these cells were polydisperse, but could be enriched ten-fold by selecting for cells with high intensity low angle (0 degrees) scatter and low intensity high angle (90 degrees) scatter. PWM gave the best enrichment (10 to 15-fold) for adult bone marrow CFCs, for CFCs from fetal sources (fetal liver, fetal blood), and for CFCs from the spleens of mice injected previously with outer membrane lipoprotein from E. coli. Three parameter sorting for CFC using the FACS (low angle scatter, high angle scatter, and PWM-fluorescence) resulted in large enrichment factors (16 to 50-fold) for CFCs from all the above sources. Over 7% of the cells sorted from bone marrow, 10% of the cells sorted from post-lipoprotein spleen, and 28% of the cells sorted from fetal peripheral blood were hemopoietic CFCs. Ninety percent of the cells in these fractions had the morphology of blast cells or myelocytes. Thus, it was possible to identify the morphological characteristics of the hemopoietic progenitor cells. Screening of other developmental systems using quantitation of fluorescence with lectins should prove of general value for the purification of selected differentiation states.

Animals↗
PubMed1980

Colony formation in agar by adult bone marrow multipotential hemopoietic cells.

Erythroid colony formation in agar cultures of CBA bone marrow cells was stimulated by the addition of pokeweed mitogen-stimulated spleen conditioned medium (SCM). Optimal colony numbers were obtained when cultures contained 20% fetal calf serum and concentrated spleen conditioned medium. By 7 days of incubation, large burst or unicentric erythroid colonies occurred at a maximum frequency of 40--50 per 10(5) bone marrow cells. In CBA mice the cells forming erythroid colonies were also present in the spleen, peripheral blood, and within individual spleen colonies. A marked strain variation was noted with CBA mice having the highest levels of erythroid colony-forming cells. In CBA mice erythroid colony-forming cells were mainly non-cycling (12.5% reduction in colony numbers after incubation with hydroxyurea or 3H-thymidine). Erythroid colony-forming cells sedimented with a peak of 4..5 mm/hr, compared with CFU-S, which sedimented at 4.25 mm/hr. The addition of erythropoietin (up to 4 units) to cultures containing SCM did not alter the number or degree of hemoglobinisation of erythroid colonies. Analysis of the total number of erythroid colony-forming cells and CFU-S in 90 individual spleen colonies gave a correlation coefficient of r = 0.93 for these two cell types. In addition to benzidine-positive erythroid cells, up to 40% of the colonies contained, in addition, varying proportions of neutrophils, macrophages, eosinophils, and megakaryocytes. Taken together with the close correlation between the numbers of CFU-S in different adult hemopoietic tissues, including individual spleen colonies, the data indicate that the erythroid colony-forming cells expressing multiple hemopoietic differentiation are members of the hemopoietic multipotential stem cell compartment.

Animals↗
PubMed1980

The responses of hemopoietic precursor cells in mice to bacterial cell-wall components.

The influence upon different cellular and humoral parameters of hemopoiesis of three structurally unrelated, highly purified bacterial cell-wall components (BCWC) was investigated. The spleens of C57BL/6 mice assayed 6 days after the injection of either lipid A or outer-membrane lipoportein, but not murein, showed a marked increase in granulocyte-macrophage, eosinophil, and megakaryocyte progenitor cell levels. The number of pluripotent hemopoietic stem cells (CFU-S) also increased in the spleens of mice treated with either lipid A or lipoprotein. Similar results were obtained following the injection of lipoprotein or lipid A into CBA or C57BL/6.nu mice. Genetically anemic Wf/Wf mice were found to have spontaneously elevated numbers of splenic progenitor cells, which increased further after the injection of lipid A. The proportions of the different splenic progenitor cell types were similar in both untreated and lipid A treated Wf/Wf mice, and in normal littermate controls. When tested in vitro, unfractionated or partially purified post-lipid A serum was found to stimulate the growth of granulocyte-macrophage progenitor cells (GM-CFC), but no detectable stimulation of eosinohphil, megakryocyte, or erythroid progenitor cells was observed. The data suggest that the rise in splenic levels of the different progenitor cells is not mediated by the corresponding types of CSF, but more likely by proliferation and differentiation of CFU-S.

Animals↗
PubMed1980

Partial priapism.

We report the third documented case of partial priapism. The proximal right corpus was distended and separated from the distral flaccid corpus by a transverse membrane. Histologic examination revealed the obstructing lesion to be owing to thrombosis of vascular erectile tissue probably secondary to trauma. Excision of this membrane promptly cured the condition.

Adult↗
PubMed1980

Radiographic heart volume, stroke volume and exercise cardiac frequency: relationship to body composition and other factors in healthy adult males.

Amongst 288 healthy male workers in heavy industry the radiographic heart volume was related to the fat-free mass and the percentage body fat which between them explained 27% of the variance. Exercise was performed on a treadmill using a belt speed of 80 m.min-1 (3 mph) and an incline which was increased progressively to 14% when the oxygen uptake was on average 61% of the maximum for these subjects. The exercise cardiac output was related to the uptake of oxygen, the fat-free mass and the ambient temperature; these factors accounted for 52% of the variance. The exercise stroke volume was related to the heart volume, the fat-free mass, the thigh muscle width, the ambient temperature, the Harvard Pack Index and other measurements which between them accounted for 42% of the variance. The exercise cardiac frequency was correlated negatively with the heart volume, the fat-free mass, the age and the Harvard Pack Index. The variates between them described 41% of the variance. The findings provide reference values for heart volume, stroke and exercise cardiac frequency in similar subjects.

Adult↗