Biull Eksp Biol Med, 1978 Aug, 86(8), 216 - 8 {Phagocytic capacity of hematopoietic tissue stromal precursor cells}; Latsinik NV et al.; A method of removal of phagocytising elements from the cell suspension by means of iron powder in magnetic field was used . Clonogenic fibroblast precursors of the hemopoietic tissue not belonging to histiocytes-macrophages, but referred to mechanocytes, were characterised by high phagocytic activity . Following Fe treatment less than 1% of clonogenic fibroblast precursors which gave rise to fibroblast colonies under conditions of monolayer cultures remained in the bone marrow cell suspension, and about 10%--in the spleen and the peritoneal exudate cell suspension. Klin Wochenschr, 1978 Aug 1, 56(15), 761 - 5 Histochemical methods for the demonstration of Thomsen-Friedenreich antigen in cell suspensions and tissue sections; Klein PJ et al.; 1 . Three different methods are described for the visualisation of the Thomsen-Friedenreich (TF) antigen on cell suspensions, formalin-fixed and paraffin embedded or frozen tissue sections: a) Rosette formation with chicken and sheep erythrocytes . b) Fluorescence-microscopy with fluorescein labelled peanut agglutinin . c) Autoradiography with 3H-labelled peanut agglutinin . 2 . The TF antigen was shown, as far as presently investigated, to be exposed on various blood cells, glomerula of the kidney and normal mammary gland after neuraminidase treatment . Mammary gland was also shown to possess TF receptors without prior treatment with neuraminidase . 3 . The exposure of this cryptantigen can be brought about by bacterial or viral neuraminidase and is followed by an antigen/antibody reaction, which can lead to possible pathological consequences. J Exp Med, 1978 Aug 1, 148(2), 580 - 91 Immunological studies of mouse decidual cells . I . Membrane markers of decidual cells in the days after implantation; Bernard O et al.; Mouse decidual cell suspensions from day 6 to day 8 of gestation were prepared by enzymatic treatment with collagenase and trypsin and tested for various membrane markers . (a) Besides H-2 antigens, Thy-1 antigens are present on about 50% of the cells; this may reflect the fibroblastic origin of decidual cells or be a marker expressed on some decidual cells possibly under hormonal control . (b) T or B lymphocytes, as defined by four Lyt antigens or surface immunoglobulins, are not present in significant amounts . (c) A substantial number of cells bearing receptors for the Fc portion of IgG (FcR) is detectable in the decidua, probably closely connected with trophoblast cells; these FcR-bearing cells may act in preventing excessive invasion of uterine tissue by trophoblast or could contribute to the protection of the embryo by interacting with maternal blocking antibodies and trophoblast . No receptors for for complement were detected, even after 16-20 h in culture after trypsin treatment. Acta Pathol Microbiol Scand {C}, 1978 Aug, 86C(4), 153 - 8 Freezing of rat lymphocytes . V . The effect of suppressor cells in phytohaemagglutinin-stimulated spleen cell cultures before and after freeze-thawing; Hem E; Spleen lymphocyte proliferation, as measured by 3H-thymidine incorporation induced by phytohaemagglutinin (PHA), was increased after freeze-thawing with 10% dimethyl sulphoxide . Depletion or intoxication of macrophages in fresh spleen cell preparations also increased lymphocyte proliferation in response to PHA . On the other hand, freezing of macrophage-depleted spleen cell suspensions lowered 3H-thymidine uptake of stimulated cultures . At concntrations above 3%, macrophages added to cultures of fresh purified lymphocytes showed a dose-dependent inhibitory effect on the PHA response, and fresh macrophages were more inhibitory than frozen-thawed macrophages . Purified lymphocytes mixed with 10% macrophages showed a higher response after freeze-thawing . It is concluded that macrophages suppress the lymphoproliferative response to PHA in rat spleen cell cultures, and that these macrophages are more sensitive than lymphocytes to the present freeze-thaw process. Br J Haematol, 1978 Aug, 39(4), 545 - 50 Evaluation of toluidine blue for measuring erythrocyte membrane loss during in vivo ageing; Greenwalt TJ et al.; A simple method using a cationic dye, toluidine blue (TB), to quantify changes of red cell membrane area has been developed and tested for its validity . After incubating a glucose-depleted red cell suspension with a fixed quantity of TB at 37 degrees C for 10 min, the remaining TB was measured spectrophotometrically at 640 nm . Using this technique, we were able to show differences in TB uptake by populations of young and old red cells . The exact mechanism for TB uptake by the red cells is not clear . Treatment of the cells with bromelin, papain and trypsin reduced the uptake of TB, but neuraminidase and ficin had little or no effect . No inhibition of TB removal by red cells was observed using heparin, D-glucose, glucuronic acid, or N-acetylglucosamine. J Natl Cancer Inst, 1978 Aug, 61(2), 319 - 25 Lung carcinoma-reactive antibodies isolated from tumor tissues and pleural effusions of lung cancer patients; Paluch E et al.; Low pH elution techniques were used on lung cancer tissues and pleural effusions of lung cancer patients to dissociate antigen-antibody complexes . The immunoglobulins obtained were assayed by indirect immunofluorescence against tissue cultures and fresh cell suspensions of various target cells; they reacted positively, in significant titers, with cells of squamous cell carcinomas and adenocarcinomas of the lung but not with cells of normal adult and fetal lung or of nonpulmonary tumors . Immunoglobulins, similarly dissociated from tumor effusions of other organs, showed no reactivity in indirect immunofluorescence tests against lung carcinoma cells. J Natl Cancer Inst, 1978 Aug, 61(2), 507 - 12 Transfer of viable putative preneoplastic hepatocytes to the livers of syngeneic host rats; Laishes BA et al.; A new approach was developed by which freshly isolated, chemical carcinogen-altered hepatocytes with the positive marker gamma-glutamyl transpeptidase (gamma-GT) could be transferred from adult donor F344 rats to the livers of syngeneic, adult host rats . Selective proliferation of gamma-GT-positive hepatocytes was induced in host rat livers, such that large, macroscopic colonies of altered hepatocytes could be generated within 10 days of the cell transfer operation . Quantitation of the number of gamma-GT-positive hepatocyte colonies (foci) appearing per square centimeter of host liver section area on day 10 following cell transfer revealed that prior treatment of host rats with a low dose of 2-acetylaminofluorene was essential for the appearance of large numbers of foci . In addition, the appearance of foci on day 10 depended on the presence of intact, gamma-GT-positive hepatocytes in the infused (transferred) cell suspensions. Steroids, 1978 Jul-Aug, 32(1), 127 - 36 Angiotensin stimulates cortisol biosynthesis in human adrenal cells in vitro; McKenna TJ et al.; Adrenal glands obtained from patients undergoing therapeutic adrenalectomy were used to study the effects of angiotensin on human adrenal steroidogenesis . It was observed that angiotensin stimulated cortisol biosynthesis . Although this has been demonstrated to occur in canine and bovine adrenals, angiotens in-induced cortisol biosynthesis has not been established in man . The possibility that angiotensin merely stimulated glomerulosa cells to secrete precursor steroids which accumulated in the medium and then diffused into fasciculata cells to provide substrate for cortisol biosynthesis was excluded by demonstrating that 3beta-hydroxy-5-pregnen-20-one (pregnenolone) and progesterone (the only pertinent precursors) did not accumulate in angiotensin-stimulated cell suspension . In addition, angiotensin stimulated cortisol biosynthesis in a fasciculata cell suspension in which angiotensin did not stimulate aldosterone production . Therefore, in human adrenal cell suspensions angiotensin appeared to act directly to stimulate cortisol synthesis by fasciculata cells . In normal subjects pre-treated with dexamethasone, angiotensin infusions failed to stimulate an increase in plasma cortisol . The physiological importance of angiotensin as a regulator of cortisol secretion remains, therefore, to be established. Cancer Lett, 1978 Jul, 5(1), 19 - 23 The effects of orally administered L-arginine HCl on the development of myeloma tumors in BABL/C mice following the injection of single cell suspensions, cell aggregates or tumor fragments; and on the growth of two ascites tumour cell lines; Pryme IF; When arginine-HCl was added to the drinking water of Balb/c mice the formation of subcutaneous tumours following the injection of single cell suspensions of MPC-11 cells was prevented, however, tumours did develop after injecting tumour fragments or aggregates of suspension-cultured cells . Arginine-HCl had no inhibitory effect upon the production of Krebs II and 6C3HED lymphoma ascites tumours in Balb/c and C3H mice respectively. Br J Cancer, 1978 Jul, 38(1), 77 - 81 Growth of human tumour cell colonies from biopsies using two soft-agar techniques; Courtenay VD et al.; Two techniques for growing colonies of human tumour cells in soft agar have been applied to cell suspensions derived from fresh tumour tissue from 48 patients . Colonies were obtained in 31 cases, with plating efficiencies between 0.01 and 15% . In 11 cases the plating efficiencies were 1% or above . There was evidence that some categories of tumour grew more readily than others under these conditions . The potential applications of the methods to clinical and experimental oncology are discussed. Transfusion, 1978 Jul-Aug, 18(4), 417 - 22 Modified microtiter tray method for blood typing; Parker JL et al.; A simple technique for red blood cell typing is described utilizing V-bottom microtiter trays with cell suspensions of 0.2% . This technique is many times more sensitive than tube or slide methods and can be used for direct agglutination, antiglobulin tests and mixed field agglutination. J Microsc, 1978 Jul, 113(2), 205 - 13 Differential leucocyte counts: a comparison of results using light and electron microscopy; Gillett R et al.; A method is described for handling leucocytes from an inflammatory peritoneal exudate prior to electron microscopy, which allows differential counts from ultra-thin sections to be made . The results of counts from ultrathin sections, viewed on the electron microscope, are compared with samples from the same cell populations prepared on a cytocentrifuge and counted by light microscopy . The results from several cell populations of widely different compositions show clearly that with suitable care taken over preparation and orientation, ultrathin sections can yield comparable differential counts to those obtained by standard light microscope procedures . Possible sources of error are discussed and the advantages of ultrastructural counting assessed . The method has a wide application wherever accurate differential counts are required from cell suspensions processed for electron microscopy. J Endocrinol, 1978 Jul, 78(1), 141 - 6 Separation of rat pituitary thyrotrophic cells; Tal E et al.; A method for the enrichment of live thyrotrophic pituitary cells is described . Pituitary glands of young male rats were removed into Earle's solution and dispersed in a 0.1% trypsin solution containing 0.5% bovine serum albumin, pH 7.2 . Nylon fibres (25 microgram) were used for the separation of the thyrotrophic cells, by stringing them across a plastic frame which fitted a plastic Petri dish containing the cell suspension . The fibres were washed with light petroleum (b.p . 60--80 degree C) and carbon tetrachloride, hydrolysed with 3 M-HCL for 30 min at room temperature and washed with distilled water and phosphate-buffered saline (pH 7.2) . The fibres were treated with thyrotrophin releasing hormone (TRH) alone or in the presence of soluble carbodiimide solution . After incubation for 1 h at room temperature, the fibres were transferred to a new Earle's medium and cells were released from the fibres by plucking them with a needle . The separated thyrotrophic cells were identified by radioimmunoassay and by electron microscopy . Using the above-mentioned methods, enrichment of thyrotrophic cells was obtained . Thus, the amounts of TSH, prolactin, LH and GH released, during 2 h of incubation, by 1.5 x 10(6) unseparated cells were 6.8 +/- 0.65, 4.1 +/- 0.47, 4.8 +/- 0.52 and 5.2 +/- 0.68 microgram respectively, while the same number of purified thyrotrophic cells released 76.1 +/- 0.42, 1.2 +/- 0.3, 0.6 +/- 0.35 and 1.6 +/- 0.22 microgram of the same hormones (means +/- S.E.M.). Immunology, 1978 Jul, 35(1), 177 - 82 Dependence of phagocytosis on strength of phagocyte-particle interaction; Capo C et al.; Sheep erythrocytes were pretreated with concanavalin A (Con-A-SRC), or glutaraldehyde (G-SRC), or specific rabbit immunoglobulin G (IgG-SRC), or specific rabbit immunoglobulin M and complement (C-SRC) . Each erythrocyte type was made to adhere to rat peritoneal cells and adhesion was measured; binding decreased as follow: conA-SRC greater than IgG-SRC greater than less than G-SRC greater than C-SRC Peritoneal cell-erythrocyte complexes were then submitted to a laminar shear flow, and resistance of binding was assayed . Binging strength decreased in the following order: G-SRC greater than C-SRC greater than IgG-SRC greater than ConA-SRC Cell suspensions were incubated at 37 degrees, and phagocytosis was measured . Ingestion decreased in the following order: G-SRC greater than IgG-SRC greater than C-SRC greater than ConA-SRC It is concluded that: Binding strength may be of importance in triggering phagocytosis; when immunocytoadherence is studied, two independent parameters should be considered: binding and binding strength . This report describes a new method that may allow discrimination between different cell subpopulations of similar binding specificities. Lab Invest, 1978 Jul, 39(1), 70 - 6 Tissue localization of T-lymphocytes by the histochemical demonstration of acid alpha-naphthyl acetate esterase; Knowles DM 2nd et al.; Nonspecific acid alpha-naphthyl acetate esterase (ANAE) activity has previously been evaluated with the sheep erythrocyte (E) rosette assay and found to be a useful T-cell marker in cell suspension studies . T-lymphocytes display a solitary red brown nodule of reaction product (T-pattern) which can be readily distinguished from the diffuse, cytoplasmic staining of monocytes (M-pattern) . Freshly obtained human tissue blocks were fixed with buffered formal sucrose and maintained in Holt's syrup; cryostat sections were cut and incubated under conditions appropriate for the histochemical demonstration of lymphocyte acid alpha-naphthyl acetate esterase activity . The results clearly demonstrate that T-pattern ANAE+ lymphocytes populate the paracortical and interfollicular regions of normal lymph nodes and tonsils and the periarteriolar sheaths of normal spleen, T-cell zones, while nearly every germinal center (B-cell area) lymphocyte is ANAE- . Macrophages and dendritic reticulum cells display diffuse cytoplasmic ANAE activity (M-pattern) . Nodular malignant lymphomas (B-cell origin) are ANAE-; an E+ diffuse, cutaneous lymphoma was ANAE+ . The histochemical demonstration of ANAE activity may represent the first reproducible, routine technique to localize T-lymphocytes in tissue sections. J Lipid Res, 1978 Jul, 19(5), 570 - 7 ACTH-induced hydrolysis of cholesteryl esters in rat adrenal cells; Vahouny GV et al.; Rat adrenal cortical cells have been prepared by collagenase dissociation of trypsin-treated adrenal tissue . The content and compositions of cholesteryl ester, phospholipid, and triglyceride fatty acids compare favorably with those of undissociated rat adrenal tissue . During 2-hour control incubations of adrenal cortical cells, steroidogenesis was not detected, and the levels of sterol ester, phospholipid, and triglyceride fatty acids were not significantly altered . Incubations with adrenocorticotropic hormone (ACTH) resulted in coricosterone production and significant depletions of sterol ester and triglyceride fatty acids, but not of phospholipid fatty acids . Although all fatty acid esters of cholesterol were hydrolyzed under these conditions, the greatest contributions to the net decrease in sterol esters were by oleate, arachidonate, and adrenate . Incubations with dibutyryl cyclic adenosine monophosphate (0.5 mM) resulted in significantly greater levels of corticosterone production than did ACTH (250 muunits), but the effects on cellular lipids were comparable to those seen with the tropic hormone . This study represents the first demonstration of hormone-induced hydrolysis of sterol esters in an in vitro cell suspension system . The results are discussed with respect to hormone-sensitive sterol ester hydrolase of adrenal cortex, and to the role of endogenous cholesteryl esters in the steroidogenic pathway. J Bacteriol, 1978 Jul, 135(1), 62 - 7 Physiological role of oxygenated cytochrome o: observations on whole-cell suspensions of Vitreoscilla; Webster DA et al.; The form of cytochrome o that predominates in Vitreoscilla cells having various levels of respiratory activity was studied by using untreated, frozen-thawed, and starved cells, which had respiratory rates decreasing in the order given . Direct spectral observation revealed that the oxygenated form of cytochrome o predominated during the aerobic steady-state oxidation of endogenous substrate or exogenous glutamate in untreated and frozen-thawed cells and was replaced by the reduced form when the cell suspensions became anaerobic . The respiratory rates, estimated inversely from the time of duration of the steady state, were correlated to the rates of oxygen consumption for the various cells . Oxidized cytochrome o predominated in aerobic starved cells . These results indicate the involvement of three forms of cytochrome o--oxidized, reduced, and oxygenated--in the catalytic and cyclic change of this cytochrome . The oxygenated form also appeared after the addition of hydrogen peroxide to the cells, but only the oxidized form appeared when ethyl hydrogen peroxide was added . The appearance of the oxygenated form with the addition of hydrogen peroxide was probably due to the reaction of the reduced cytochrome with the oxygen that had evolved by the action of catalase present in the cells. Stain Technol, 1978 Jul, 53(4), 199 - 204 A permanent cell viability assay using alcian blue; Thornthwaite JT et al.; The alcian blue dye exclusion method for glutaraldehyde-fixed cells has been utilized with "centrifugal cytology" to prepare permanent records of the viability of individual cells present in suspensions . The viability of spleen cell suspensions separated by linear bovine serum albumin density gradient centrifugation has been measured with this method . Combined light and scanning electron microscopy of nonviable and viable cells demonstrated membrane alterations in alcian blue-stained nonviable cells, while viable cells were spherical and displayed uniform surface features. Brain Res, 1978 Jun 16, 148(2), 313 - 31 Separation of cell types from the developing cerebellum; Cohen J et al.; The heterogenous population of perikarya (cells) obtained by dissociating cerebellar tissue of developing postnatal rats was separated by sedimentation at unit gravity . Peak fractions, defined by monitoring the distribution of different size classes with a particle analyzer, were enriched in ultrastructurally well-preserved and metabolically competent specific cell types . These fractions included the peak of rapidly sedimenting cells comprising large neurones, such as Purkinje cells, which accounted for about 50% of the cells (vs . 2% in the initial cell suspension) and for a much greater proportion of the total cell mass (over 80%) . More slowly sedimenting peak fractions contained 2-5-fold enrichments in replicating cells (assessed in terms of {3H}thymidine incorporation into DNA or by the frequency of mitotic cells), astroglia-like cells and external and differentiating granule cells, respectively . Separated, replicating cells continued synthesizing DNA in vitro; the {3H}thymidine incorporation rate was about 5-fold greater in the fraction enriched in proliferating cells than in the total cell suspension . Besides their structural integrity, the viability of the cells was also indicated by the finding that the proportion of trypan blue-excluding cells in all fractions exceeded 80% . Moreover, protein synthesis, in terms of incorporation of labelled amino acids, continued in the separated cells at a linear rate for a relatively long time . The rate per cell was highest in the large neuronal fraction, and lowest in the astroglia-like fraction. Cytobiologie, 1978 Jun, 17(1), 182 - 96 Penetration and entrapment of large particles in erythrocytes by electrical breakdown techniques; Vienken J et al.; Human erythrocytes suspended in isotonic solutions were subjected to haemolysis by application of an electric field pulse to the cell suspension . The field strengths used were 12 and 16kV/cm, respectively; the pulse duration 40 microseconds . The lysed cells showed resealing properties . The permeability change of the membrane generated by the field pulse and by the subsequent osmotic processes were large enough to facilitate the penetration and entrapment of ferritin and Latex particles (diameter: 0.091 and 0.176 micron, respectively) as revealed by electron microscopy . Correct identification of the Latex particles in the electron-micrographs indicated that LOYTER et al . {J . Cell Biol . 66, 292 (1975)}, who recently demonstrated the entrapment of Latex spheres in erythrocytes prepared by osmotic haemolysis mistook electron-dense bodies probably consisting of denaturated protein for Latex particles . Under conditions of osmotic haemolysis, carried out according to BODEMANN and PASSOW, particles could only occasionally be detected within the membrane itself and never within the cell interior, suggesting that the electrical haemolysis method is much more effective in the generation of large holes in the membrane. Br J Cancer, 1978 Jun, 37(6), 1015 - 9 A soft-agar procedure measuring growth of human colonic carcinomas; Kimball PM et al.; Cell suspensions from 5 human colonic carcinomas were fractionated by velocity sedimentation and plated in soft agar . Cluster formation was restricted to the purest fraction of epithelial cells, as had been determined by immuno- and histochemical criteria . Plating efficiencies for the 5 specimens were 1.0-4.5% . The effects of varying the incubation period and inoculum size upon growth were studied using unseparated cell suspensions from 6 specimens . Clusters were apparent after 3 weeks in culture, and maximum cluster formation was typically seen by 5 weeks . Cluster formation appeared concentration-dependent, and individual specimens varied with respect to the inoculum most conducive to growth . The maximum plating efficiencies for unseparated cells were unseparated cells were 0.4-1.7%. J Immunol, 1978 Jun, 120(6), 1876 - 80 In situ lymphoid cells of mouse mammary tumors . I . Development and evaluation of a method for the separation of lymphoid cells from mouse mammary tumors; Blazar BA et al.; A method of separating lymphoid cells from solid mouse mammary tumors was developed and evaluated . In this method the tumors are digested with 0.01% collagenase, 0.01% DNAase, and 0.025% trypsin in Dulbecco's PBS into suspensions of cells with a viability of 90% . The suspensions are fractionated on a continuous gradient of Ficoll in tissue culture medium . In model experiments this gradient was found to separate, cleanly, admixed cells of an established mammary tumor cell line and dissociated thymus glands . Recovery rates were 50% for the tumor cells and 80% for the thymocytes . The preparation of the cell suspensions and the gradient separation procedure are not harmful to the cells as indicated by trypan blue exclusion and the ability to grow in cell culture. Cancer Res, 1978 Jun, 38(6), 1539 - 49 Ultrastructure of adult rat hepatocytes cultured on floating collagen membranes; Sattler CA et al.; The ultrastructure of primary hepatocytes cultured for 2 to 17 days on floating collagen membrane was evaluated . A pellet of the hepatic cell suspension used to inoculate the collagen membranes contained some single cells and many aggregates of two, three, or four cells . Desmosomes were split during the perfusion, but tight junctions and gap junctions remained intact . By 2 days in culture, the hepatocytes had formed a monolayer of cells of polygonal shape with newly synthesized desmosomes between cells . Since the flexible floating collagen membrane decreases in size as the monolayer forms, the hepatocytes do not flatten out, as is characteristic of cells cultured on a rigid substrate . Hepatocytes in culture for 10 days or less exhibited large lamellar arrays of rough endoplasmic reticulum, well developed Golgi complexes, and structures resembling bile canaliculi, which possess tight junctions and desmosomes separating them from the intercellular space . Microfilaments oriented parallel to the plasma membranes of adjoining cells and in an intermeshed network at the edge of the monolayer and beneath the plasma membrane bordering the medium increased in size and number in older cultures . After 17 days in culture, the cells maintained tight junctions, desmosomes, Golgi complexes, and rough endoplasmic reticulum in small lamellar stacks and in small vesicles . Since hepatocytes on the floating collagen membrane retain most of the subcellular structural elements characteristic of normally functioning hepatocytes for 2.5 weeks, this system may be valuable for future experiments involving drug metabolism and carcinogenesis in vitro. Clin Exp Immunol, 1978 Jun, 32(3), 554 - 62 Inhibition of polyclonal B-cell activation by suppressor monocytes in patients with sarcoidosis; Katz P et al.; The present study employed a direct plaque-forming cell (PGC) assay following pokeweed mitogen (PWM) induced polyclonal activation of B lymphocytes in sarcoidosis to evaluate the in vitro humoral immune response in this disease and to delineate the immunoregulation of this response . Sarcoidosis lymphocytes had a suppressed PFC response to polyclonal activation, but were unable to suppress normal B-cell PFC responses in allogeneic co-cultures . Removal of a cell type, which was corticosteroid-resistant, radio-resistant, adherent and a non-T cell, from sarcoidosis mononuclear cell suspensions reversed the suppressed PFC response, indicating the presence of a suppressor monocyte . Thus in vitro suppressor cell activity has now been demonstrated in this disease, which is characterized by multiple immunological aberrancies. Cancer, 1978 Jun, 41(6), 2183 - 91 Thymoma: an immunologic and electron microscopic study; Cossman J et al.; Ultrastructural features, surface morphology and immunologic surface markers were examined on the cells of three human thymomas . The vast majority of the lymphocytes from the thymomas formed spontaneous rosettes with unsensitized sheep erythrocytes in both cell suspension and frozen tissue section and were, therefore, T cells . In addition to the lymphocytes, epithelial cells and macrophages were observed within the thymomas by transmission electron microscopy . When examined by scanning electron microscopy, most lymphocytes had virtually smooth surfaces, whereas cells believed to be epithelial in origin had surface projections. Can J Biochem, 1978 Jun, 56(6), 430 - 9 Preparation of cell populations with stabilized erythropoietic potential from the primitive streak chick blastodisc: some implications for control of gastrulation; Wainwright SD et al.; Improved methods for preparation from primitive streak chick blastodiscs of cell suspensions capable of forming erythroid cells in culture have been developed . When blastodiscs were preincubayed with hyaluronidase in the absence of collagenase before cell dispersion and a high concentration of methyl-alpha-mannoside was present in all media, the yields of cells were some 10-fold higher than those obtained by former procedures . Cell suspensions obtained consisted almost entirely of viable cells, yielded large numbers of free mature erythrocytes in liquid culture, and formed erythroid colonies and bursts in solidified medium . The capacity to form differentiated cells after resedimenrtation through Ficoll density gradients was partly stabilized . Addition of gee yolk homogenate to the blastodiscs immediately following treatment with hyaluronidase and to all media used thereafter largely stabilized the capacity to form erythroid cells during resedimentation through Ficoll density gradients . Possible relevance of observations made during development of the procedures to the control of onset of cell migration in the process of gastrulation is indicated. J Bacteriol, 1978 Jun, 134(3), 718 - 27 Oxidation kinetics and chemostat growth kinetics of Thiobacillus ferrooxidans on tetrathionate and thiosulfate; Eccleston M et al.; Growth of Thiobacillus ferrooxidans in batch culture on 10 mM potassium tetrathionate was optimal at pH 2.5 (specific growth rate, 0.092 h-1) . Oxygen electrode studies on resting cell suspensions showed that the apparent Km for tetrathionate oxidation (0.13 to 8.33 mM) was pH dependent, suggesting higher substrate affinity at higher pH . Conversely, oxidation rates were greatest at low pH . High substrate concentrations (7.7 to 77 mM) did not affect maximum oxidation rates at pH 3.0, but produced substrate inhibition at other pH values . Tetrathionate-grown cell suspensions also oxidized thiosulfate at pH 2.0 to 4.0 . Apparent Km values (1.2 to 25 mM) were of the same order as for tetrathionate, but kinetics were complex . Continuous culture on growth-limiting tetrathionate at pH 2.5, followed by continuous culture on growth-limiting thiosulfate at pH 2.5, indicated true growth yield values (grams {dry weight} per gram-molecule of substrate) of 12.2 and 7.5, and maintenance coefficient values (millimoles of substrate per gram {dry weight) of organisms per hour) of 1.01 and 0.97 for tetrathionate and thiosulfate, respectively . Yield was increased on both media at low dilution rates by increase in CO2 supply . The apparent maintenance coefficient was lowered without affecting YG, suggesting better energy coupling in CO2-rich environments . Prolonged continuous cultivation on tetrathionate or thiosulfate did not affect the ability of the organism to grow subsequently in ferrous iron medium. Virchows Arch B Cell Pathol, 1978 May 19, 27(3), 205 - 15 Regenerative proliferation of mouse epidermal cells following application of a carcinogenic skin irritant (MCA) . Micro-flow fluorometric DNA measurements and 3H-TdR incorporation studies of isolated basal cells; Clausen OP; 0.025 ml of a 1% solution of the complete skin carcinogen 20-methylcholanthrene (MCA) dissolved in benzene was applied to the back skin of hairless mice . At different time intervals up to 3 days after the carcinogen application groups of animals were injected i.p . with 30 muCi 3H-TdR 30 min before they were killed . Single cell suspensions of epidermal basal cells were prepared by a combined enzymatic and mechanical separation method, and the DNA frequency distribution pattern from each cell suspension was measured by means of micro-flow fluorometry . Smears for autoradiography were made from each cell suspension and the labeling index and mean grain count assessed . After a short initial delay, MCA induced an increase in the labeling index similar to that observed after non-specific cell injury and cell loss . Thereafter, the cells were considerably delayed in their progression through the S phase, with a low exit from S resulting in a transient emptying of the G2 compartment, without indications of any significant delay of the passage through G2 phase . The cells that had been injured by the MCA application in or just before S phase proceeded into the G2 phase and mitosis more than 24 h after the initiation of DNA synthesis . The cell kinetic reaction of epidermis to a single application of MCA is thus very different from that caused by a nonspecific cell damage, e.g . application of the vesicant agent cantharidin or removal of surface cells by cellophane tape stripping. Experientia, 1978 May 15, 34(5), 602 - 3 Metabolic studies of Hg-203 on Chlamydomonas reinhardi; Macka W et al.; Sterile cultures of Chlamydomonas reinhardi, WT+, were treated with Hg-203 at 25 degrees C to identify probably formed volatile mercury compounds . Experiments were performed with living and dead cells under aerobic or anaerobic conditions, respectively, and the mercury concentration was measured in the system algae/nutrient medium . We found a timerelated decrease of mercury concentration in the cell suspension and the cell-free nutrient medium due to a reduction of Hg++ to Hg0, probably caused by extracellular enzymes; monomethyl or dimethyl mercury could not be detected. J Biol Chem, 1978 May 10, 253(9), 2902 - 4 The induction of ferritin synthesis in circulating larval red blood cells; Theil EC; The circulating red blood cells formed in bullfrog larvae, chicken embryos, and mouse embryos contain large amounts of ferritin and storage iron in excess of the need for hemoglobin . In contrast, the circulating red cells of adult animals contain little ferritin . Ferritin synthesis and iron storage are coordinated with differentiation and hemoglobin synthesis in the red cells of adults . In order to test the hypothesis that ferritin synthesis could be controlled independently of hemoglobin synthesis and differentiation in the red cells formed early in life, bullfrog larvae were injected with iron to determine if ferritin synthesis was increased in the circulating red cells . Within 17 h after the injection of iron, the synthesis of ferritin, assayed as the incorporation of {14C}leucine by cell suspensions prepared from circulating red cells, was increased from 2.9 to 10.2% of the total protein, and the specific activity of the ferritin synthesized increased from 1100 to 3000 cpm/A280 . There was no change in the hematocrit of the animals nor in the specific activity of hemoglobin synthesized by suspensions of red cells (average, 720 cpm/A280) . The results suggest that in mature, larval red cells, ferritin synthesis can be controlled by changes in the extracellular environment . The results also indicate that ferritin synthesis can be controlled independently of hemoglobin synthesis with which it is coordinated during erythroid differentiation in adult animals. J Pathol, 1978 May, 125(1), 53 - 8 Macrophages in giant cell tumours of bone; Wood GW et al.; Five giant cell tumours of bone were studied to determine the degree of macrophage infiltration and whether the giant cells expressed the characteristics commonly associated with macrophages, i.e., IgGFc and C3 receptors, phagocytosis and non-specific esterase activity . Macrophages were assessed in trypsin-derived tumour cell suspensions by IgGEAC rosette formation and in frozen sections of tumour by EA adsorption . The percentage of macrophages in cell suspensions from four of the tumours ranged from 11 to 40 per cent . Strong EA adsorption occurred over 35 to 95 per cent . of the tumours' surface and significant non-specific esterase positivity was observed in the tumour sections . The giant cells were receptor negative and non-phagocytic, but a low percentage of them expressed esterase activity . The results strongly suggest that despite the fact that large numbers of macrophages were present in the tumours, the giant cells were derived from cells other than macrophages. Eur J Immunol, 1978 May, 8(5), 331 - 5 Primary in vitro antibody formation in the rat: partial characterization and properties of an inhibitor cell present in normal spleen; Corvalan JR et al.; Rat spleen cells are shown to be unresponsive to sheep red cells (SRBC) in vitro under conditions in which thoracic duct lymphocytes (TDL) respond very well . By adding unresponsive spleen cells to responsive TDL cultures, the spleen cells are shown to contain an inhibitor capable of preventing the response to SRBC . The inhibitory activity is a property of live cells; it is sensitive to radiation doses as low as 100 R x rays and to mitomycin C . It can be completely removed from spleen cell suspensions by extraction with large amounts of carbonyl iron or by filtration through nylon wool columns . It is less efficiently removed by filtration through Sephades G-10 columns, and is completely resistant to the cytotoxic effects of silica . From a practical point of view, extraction of a spleen cell suspension with carbonyl iron is a useful method of obtaining fully responsive lymphocyte populations from rat spleen. Acta Pathol Jpn, 1978 May, 28(3), 435 - 44 Growth and cytogenetic characteristics of nickel sulphide-induced rhabdomyosarcomas in rats; Yamashiro S et al.; Rhabdomyosarcomas induced by single intramuscular injections of nickel sulphide (Ni3S2) in Fischer and Hooded rats were cultured in vivo and in vitro to study their growth characteristics and chromosomal constitution . The tumor cell suspensions cultured in vitro exhibited more myogenic differentiation on the coverslips than those cells grown in vivo in diffusion chambers . A characteristic feature of in vivo cultures was the appearance of microclusters which resembled the primary tumors . Chromosome analyses of primary tumors revealed that a majority of these had a modal number in the diploid or near diploid range . Fischer rat primary tumor cells exhibited abnormal configurations including rings, dicentrics and triradials . A comparison of the chromosome make-up of the primary tumors and their metastases was performed on four sets of tumors . Three out of four metastases examined showed the diploid chromosome make-up characteristic of the primary tumors suggesting that the tumors with the diploid or near diploid chromosome constitution are more likely to produce metastases. J Immunol, 1978 May, 120(5), 1550 - 3 Normal mouse serum immunosuppressive activity: action on adherent cells; Martineau RS et al.; The immunosuppressive activity of normal mouse serum (NMS) was evaluated by enumeration of primary in vitro plaque-forming cell response produced by normal spleen cell suspensions in response to sheep erythrocytes . The site of action of the inhibitor(s) was shown to be at the level of adherent cells since incubation of this cell type with NMS could reproducibly inhibit responses, whereas, in appropriate experiments, incubation of nonadherent cells with NMS had no effect . Furthermore, addition to normal spleen cells of excess spleen-adherent cells, normal peritoneal adherent cells, or of 2-mercaptoethanol could abrogate the inhibitory ability of NMS . Addition of excess nonadherent cells under the same conditions had no influence on inhibition . NMS was also shown to decrease the number of cell clusters, hemolytic clusters, and the number of nucleated cells recovered upon culture termination . It is suggested that NMS, acting on the adherent cell may hinder cell-to-cell contact and interactions. Biophys J, 1978 May, 22(2), 171 - 8 High gradient magnetic separation of erythrocytes; Owen CS; The high gradient magnetic separation technique has been applied to separate paramagnetic erythrocytes from a cell suspension that also contained diamagnetic cells . Paramagnetism was induced in the red blood cells by oxidizing the iron atoms in the cell hemoglobin to the ferric state (methemoglobin) . Diamagnetic cells were either untreated erythrocytes, containing oxyferrohemoglobin, or leukocytes in a suspension of mouse spleen cells . Cell suspensions were passed through a column containing 40 micron diameter stainless steel wire in a high magnetic field (33 kG) . The paramagnetic cells were retained on the surface of the wire while the diamagnetic cells passed through . Elution of the paramagnetic cells was accomplished by removing the column from the magnet, in effect turning off the field. Br J Cancer, 1978 May, 37(5), 806 - 17 Natural cytotoxicity of haemopoietic cell populations against murine lymphoid tumours; Burton RC et al.; Homozygous nude and normal mice of 3 strains, BALB/c, CBA and C57BL, were used as sources of nucleated haemopoietic "natural killer" (NK) cells . These killer cells could lyse a wide range of syngeneic and allogeneic lymphoid tumour cell lines in vitro, and it was found that cell suspensions from nude mice were always significantly more active than those from normal mice, and that the most active effector population was a polymorph-enriched peritoneal-exudate cell suspension . Eosinophils did not appear to be involved in the phenomenon, and mononuclear peritoneal-exudate cell suspensions were actually highly inhibitory . Three non-lymphoid tumours, a carcinoma, a fibrosarcoma and a mastocytoma, were totally resistant to in vitro lysis . Although all susceptible tumour cell lines were C-type virus-associated, not all of these tumours were killed by all strain sources of spleen cells, indicating a specificity of killing. Br J Cancer, 1978 May, 37(5), 718 - 22 Stability of transplanted murine tumour systems after storage of cells at -196 degrees C for up to 13 years; Hewitt HB et al.; Two murine lymphomas of spontaneous origin were stored in liquid N2 as cell suspensions in DMSO for 8 and 13 years respectively . Isogenic transplantation assays done some years before freezing and immediately after thawing indicated no measurable loss of clonogenic cells during freezing, storage or thawing; the number of cells required for 50% successful transplantation remained close to unity in both cases . The overall data revealed no evidence of an alteration of the receptivity of the mouse colonies over a period of 13 or 20 years . We attribute this remarkable stability to close supervision of the systems within a single laboratory. J Clin Pathol, 1978 May, 31(5), 461 - 8 Cell receptor studies on six anaplastic tumours of the thyroid; Macaulay RA et al.; Cell suspensions from six anaplastic thyroid tumours were studied for expression of lymphocyte and macrophage surface markers, and results were correlated with electron microscopy, clinical extent of disease, and response to radiotherapy . The clinical presentation of the disease was similar in all six patients . In five cases, many of the cells showed surface immunoglobulin . Electron microscopy was available on three of these and showed appearances in keeping with malignant lymphoma . The single case whose cells did not show surface immunoglobulin had an entirely different ultrastructure and was probably a carcinoma . This case was the only one that did not achieve complete remission with radiotherapy and the patient died from extensive local recurrence . It is concluded that receptor techniques are of value in distinguishing between malignant lymphoma and other anaplastic tumours of the thyroid, particularly when the results are correlated with histology. Cancer, 1978 May, 41(5), 1845 - 56 Cytology and colchicine sensitivity of viable cells from lymph nodes with malignant lymphoma; Schrek R et al.; Viable cell suspensions were prepared from 31 nodes diagnosed non-Hodgkin's malignant lymphoma, and from 30 non-malignant nodes . The cells were examined and counted by phase contrast microscopy . The suspensions were characterized by the percentage of large cells and by a colchicine-sensitivity index . The finding of more than 6% large cells or the finding of a sensitivity index of more than 30% was considered a positive test for a malignant lymphoma . According to these criteria there were 2 false positives in 30 reactive nodes and one false negative in 31 malignant nodes . Findings on 3 nodes diagnosed angioimmunoblastic lymphadenopathy suggested malignancy . The colchicinesensitivity index of blood lymphocytes seemed useful for monitoring lymphoma patients for leukemic involvement. Am J Med, 1978 May, 64(5), 788 - 94 Terminal deoxynucleotidyl transferase in the diagnosis of leukemia and malignant lymphoma; Kung PC et al.; Neoplastic cells from 253 patients with leukemia and 46 patients with malignant lymphoma were studied for the presence of terminal deoxynucleotidyl transferase (TdT) by biochemical and fluorescent antibody technics . TdT was detected in circulating blast cells from 73 of 77 patients with acute lymphoblastic leukemia, 24 of 72 patients with chronic myelogenous leukemia examined during the blastic phase of the disorder and in cell suspensions of lymph nodes from nine of nine patients with diffuse lymphoblastic lymphoma . Blast cells from six of 10 patients with acute undifferentiated leukemia were TdT positive, but the enzyme was found in only two of 55 patients with acute myeloblastic leukemia . TdT was not detected in other lymphocytic or granulocytic leukemias or in other types of malignant lymphomas . The fluorescent antibody assay for TdT permits rapid and specific identification of the enzyme in single cells . The TdT assay is clinically useful in confirming the diagnosis of acute lymphoblastic leukemia, evaluating patients with blastic chronic myelogenous leukemia, and distinguishing patients with lymphoblastic lymphoma, whose natural history includes rapid extranodal dissemination, from patients with other poorly differentiated malignant lymphomas. Biomedicine, 1978 May-Jun, 28(3), 156 - 60 Production of stem cell proliferation stimulators and inhibitors by haemopoietic cell suspensions; Wright EG et al.; In mice treated with phenylhydrazine haemopoietic spleen colony forming cells (CFU-S) are proliferating rapidly in the bone marrow but not in the spleen . Using such mice we have investigated the production of factors responsible for the control of CFU-S proliferation . When irradiated spleen cells are incubated with non-irradiated bone marrow cells there is a marked fall in the proportion of femoral CFU-S in DNA synthesis . In the converse experiments, rapid triggering of splenic CFU-S is achieved . Both these effects can be eliminated by washing the irradiated cells prior to incubation; they are, however, retained in the supernatant media "conditioned" by these cells . When the washed cells are incubated in fresh medium at 37 degrees C both stimulatory and inhibitory activities reappear but after different incubation periods . The data demonstrate that both proliferation stimulatory and inhibitory factors acting on CFU-S can be produced by the same haemopoietic cell suspension. Transfusion, 1978 May-Jun, 18(3), 353 - 5 Autoagglutination dispersal utilizing sulphydryl compounds; Reid ME; Autoagglutination caused by powerful IgM cold autoagglutinins can be dispersed by sulphydryl compounds to provide an unagglutinated red blood cell suspension suitable for immunohematologic testing . For this 0.1 M 2-mercaptoethanol is added to twice washed red blood cells . Following agitation for 10 minutes at 37 C an unagglutinated red blood cell suspension is obtained without adverse effect to red blood cell antigens and without removal of in vivo bound IgG or complement . Dithiothreital may be substituted for 2-mercaptoethanol if the incubation time is increased to 15 minutes. Clin Exp Pharmacol Physiol, 1978 May-Jun, 5(3), 223 - 7 The effect of various drugs on adenosine triphosphate content and histamine release from rat peritoneal cell suspensions rich in mast cells; Day RO et al.; 1 . Iopanoic acid and iophenoxic acid were potent inhibitors of compound 48/80-induced histamine release from rat peritoneal cell suspensions rich in mast cells and caused a parallel dose-related reduction in ATP content of these cells . 2 . Lignocaine was less potent and ouabain and probenecid were ineffective in inhibiting histamine release and reducing cellular ATP content . 3 . Various drugs can inhibit 48/80-induced histamine release from rat peritoneal cells and this activity may result from depletion of cellular ATP . It is apparent that little structural specificity is required for activity in this cell system. Appl Environ Microbiol, 1978 May, 35(5), 858 - 62 Inhibition of light-induced pH increase and O2 evolution of marine microalgae by water-soluble components of crude and refined oils; Armstrong JE et al.; Light-induced alkalinization of the extracellular medium was found to be a common feature of the primary photosynthetic process of several marine microalgae . The light-induce PH increase of suspensions of whole cells was immediately and severely inhibited by a single dose of water-soluble components from crude and fuel oils . Differential effects on the rates of microalgal photosynthetic O2 evolution and cell suspension pH increase suggest different toxicity mechanisms of the water-soluble components of no . 2 fuel oil as compared with Southern Louisiana and Jay Crude oils . These short-term studies on the nature of sublethal petroleum toxicity to microalgae indicate that the primary effect may be through direct action on the energy-yielding electron transport systems. Clin Exp Immunol, 1978 Apr, 32(1), 186 - 91 Comparison of the relative cytotoxic effector cell capabilities and the proportions of cells bearing various surface markers in human tonsil and peripheral blood mononuclear cells; Hunninghake GW et al.; The relative cytotoxic effector cell capabilities and the proportions of cells bearing various surface markers in human tonsil and peripheral blood mononuclear cells has been studied . The peripheral blood contained a substantial proportion of monocytes (22 +/- 2.9%) compared to tonsil cell suspensions (2.5 +/- 0.3%) . The percentages of T lymphocytes was significantly higher in the blood than in the tonsil (P is less than 0.01); however, the percentages of cells forming rosettes with 7S EA were not significantly different in each group (P greater than 0.5) . Mitogen-induced cellular cytotoxicity by blood and tonsil mononuclear cells against Chang cells was proportional to the percentages of T lymphocytes in these cell suspensions, and both antibody-dependent and mitogen-induced cellular cytoxicity against sheep red blood cells was proportional to the percentages of monocytes in these suspensions . Tonsil mononuclear cell suspensions were incapable of mediating antibody-dependent cellular cytotoxicity against Chang cells, whereas blood mononuclear cells functioned normally . These findings are in contrast to the findings of similar percentages of Fc receptor-positive lymphocytes in blood and tonsil mononuclear cell suspensions . Previous studies have shown that the effector cells against antibody-coated Chang cells are Fc receptor-positive lymphocytes . These studies show that in the case of cytotoxicity mediated by an Fc receptor-bearing lymphoid cell, there may be a clear discrepancy between the relative proportions of Fc-bearing lymphoid cells in different organs and the relative levels of cytotoxicity. Strahlentherapie, 1978 Apr, 154(4), 277 - 81 {Short-term incubation in vitro with precursors of nucleic acids on human primary tumors and metastases of carcinoma of the breast (author's transl)}; Kaufmann M et al.; A technique of short-term tests in vitro by means of the incubation of tumor cell suspensions is utilized as a radioactive-biochemical method for pretherapeutic determination of the resistance in human cancers of the breast . Cell suspensions from primary tumors and metastases reveal individually different responses to cytostatics in vitro . It is possible, therewith, to differentiate two tumor collectives related to in vivo resistant or in vivo sensitive tumors . The responses of the primary lesion and the axillary lymphatic metastasis of the same carcinoma may in single cases also differ in vitro, according to clinical experience with the therapy of breast cancer . A distinct relation can be shown between the histological type of a carcinoma and its in vitro capacity of resistance. J Clin Microbiol, 1978 Apr, 7(4), 361 - 7 Peptocoagulase: clotting factor produced by bovine strains of Peptococcus indolicus; Switalski LM et al.; The production of a clotting factor (peptocoagulase) by bovine clinical isolates of Peptococcus indolicus and its nature were investigated . Extracellular peptocoagulase was demonstrated in culture filtrates of 93% and cell associated with washed cell suspensions of 100% of the 75 isolates tested . Both citrated and heparinized plasma were clotted . Crude peptocoagulase was nondialyzable, precipitated with (NH4)2SO4 at 40% saturation, somewhat resistant to heating at both neutral and acid pH, and chloroform insoluble . Culture filtrate did not contain proteolytic activity with albumin and casein, as substrates and no esterase activity was detected with tosylarginine and benzoylarginine methyl esters as substrates . The clotting reaction required peptocoagulase, prothrombin, and fibrinogen . The activation of prothrombin appeared to involve a stoichiometric reaction with peptocoagulase, possibly by formation of a stable complex. Biull Eksp Biol Med, 1978 Apr, 85(4), 451 - 4 {Heterologous antisera to stromal mechanocytes of the bone marrow}; Ivanov-Smolenskii AA et al.; Heterologous rabbit antifibroblast serum (AFS) to stromal fibroblasts of the guinea-pig bone marrow was obtained . AFS was shown to bind specifically stromal fibroblasts and their precursors (but not macrophages) in the monolayer primary cultures of the bone marrow, thymus and spleen of guinea pigs (in the complement-dependent cytotoxic reaction and in the indirect immunofluorescence test); AFS also bound precursors of blood fibroblasts and peritoneal exudate (in the cytotoxic reaction) . Precursors of the thymus fibroblasts were found to be more sensitive to the AFS action than the spleen and bone marrow precursors, this suggesting that the thymic stromal mechanocytes had a greater concentration of common tissue-specific antigens on their surface . Precursors of the stromal fibroblasts in native cell suspensions were found to be essentially more sensitive to the cytotoxic action of AFS than the colony-forming fibroblasts on the passage cultures. Immunology, 1978 Apr, 34(4), 715 - 20 Activation of B lymphocytes by mycoplasma mitogen(s); Naot Y et al.; Various strains of the murine mycoplasma M . neurolyticum have been shown to induce extensive blast transformation of mouse lymphocytes, comparable in strength to mitogenicity exerted by these mycoplasma species on rat lymphocytes . The data summarized in this report demonstrate that this mitogenic effect is non-specific . Lymphoid cells from mycoplasma free, germ-free mice were activated to the same extent as those lymphocytes obtained from conventionally bred animals . Lymph node cell suspensions obtained from athymic nude mice were strongly activated by M . neurolyticum mitogen . Furthermore, mouse thymocytes and mouse T-cell enriched populations, were not stimulated by these mitogens . It was thus suggested that M . neurolyticum activates mouse B lymphocytes. Endocrinology, 1978 Apr, 102(4), 1053 - 60 Decreased Leydig cell responsiveness in the testicular feminized male rat; Purvis K et al.; The Stanley-Gumbreck pseudohermaphrodite or testicular feminized male (tfm) rat exhibits a decreased Leydig cell sensitivity to human CG (hCG) measured by androgen and cyclic-3',5',-adenosine monophosphate production in vitro . These changes were associated with an 80% reduction in the number of LH receptors in the tfm testis, when compared on the basis of equivalent amounts of testis particle protein or per 10(6) isolated Leydig cells . Androstenedione and not testosterone is the major androgen secreted by the tfm Leydig cell and androstenedione secretion is, therefore, a more appropriate end point than testosterone secretion for Leydig cell function in tfm animals . A dose of hCG (3 ng/2 ml) which elicited a near maximal response in androgen production from the decapsulated testes and Leydig cell suspensions of normals rats, did not significantly stimulate androgen production from Leydig cells of the tfm animals . A much higher dose of hCG (200 ng/2 ml) gave a response from the tfm Leydig cells which was comparable to that obtained with 3 ng from Leydig cells of normal littermates . This indicates that the small number of LH receptors on the tfm Leydig cell membrane are functional and that the reduction in receptor number results in a decrease in the sensitivity of response to LH rather than a reduction in the maximum steroid response. J Appl Physiol, 1978 Apr, 44(4), 542 - 6 Physical activity and hypophysectomy on the aerobic capacity of ligaments and tendons; Vailas AC et al.; Traditionally, ligaments and tendons (L and T) have been regarded as metabolically inert structures . However, sufficient biochemical evidence on the metabolism of collagen has indicated that such a concept is no longer tenable . To determine whether L and T respond to increased or decreased levels of chronic exercise, studies were undertaken to measure their aerobic capacities . For comparative purposes, similar measurements were obtained from liver and skeletal muscles secured from normal and hypophysectomized male rats . Oxygen consumption and cytochrome oxidase (CO) activity was recorded from cell suspensions that had been prepared with the inclusion of collagenase and with elastase added to the medium . The O2 results showed that L and T had values that were approximately 10 times lower than liver tissue and 7.5 times less than the means from skeletal muscles . Hypophysectomy caused marked reductions in O2 uptake of liver and muscle tissues; but had no impact on L and T . When CO activity of these connective tissues were evaluated, immobilization and hypophysectomy caused significant reductions that ranged from -36% to -59% respectively . Training, on the other hand, resulted in increases of less than 10% in the activity of this enzyme within L and T while being elevated in muscle tissue by 58% . It was concluded that the metabolic activity of L and T was lowered with decreased levels of physical activity but it was unclear why chronic exercise did not produce the opposite effect. Cancer Res, 1978 Apr, 38(4), 939 - 41 Prevalence of non-T-cells in the replication of the N-tropic, type C virus of young AKR mice; Gisselbrecht S et al.; The XC infectious center assay was used to study the nature of the lymphoid cells producing N-tropic C-type viruses in preleukemic AKR mice . Viral production by thymic cell suspensions was very low and was possibly due to contaminating cells . Production at least 100-fold higher was found in spleen cells and was probably due to non-T-cells . The significance of these results is discussed briefly, including the possibility that the N-tropic XC syncitia-inducing type C virus of young AKR mice is not the leukemogenic agent. Mech Ageing Dev, 1978 Apr, 7(4), 309 - 20 Cyclic nucleotide levels in resting and mitogen-stimulated spleen cell suspensions from young and old mice; Tam CF et al.; The levels of cyclic adenosine 3', 5'-monophosphate (cAMP) in suspensions of unstimulated spleen cells from tumor-free 30-month old (C57BL/10Sn X C3H/HeDiSn)F1 hybrid mice averaged only 14% of that of 6-month old mice . By contrast, the level of cyclic guanosine 3', 5'-monophosphate (cGMP) in spleen cell suspensions from old mice was about 270% that of young mice . The cAMP/cGMP ratio for the unstimulated (resting) state showed a decline by 30 months to about 5% of its 6-month value . Cyclic nucleotide levels were also measured in cell suspensions from old and young mice at intervals over a two hour period following in vitro stimulation with the plant mitogens phytohemagglutinin, concanavalin-A and pokeweed mitogen . Quantitative and in some instances qualitative differences in responses were noted . These results might conceivably reflect either age-related changes in the splenic lymphoid cell subpopulations or intrinsic cellular alterations or both . It is unlikely that changes of this degree could be wholly explained by population shifts . An imbalance in cyclic nucleotide levels in both resting and stimulated lymphoid cells in older animals might contribute to the immune dysfunction known to occur with normal aging. Br J Cancer, 1978 Apr, 37(4), 530 - 5 Measurement of H-2 antigen and immunogenicity of methylcholanthrene-induced murine sarcomas; Pearson T et al.; For each of a set of 11 methylcholanthrene-induced sarcomas of B10 mice, we measured the amount of H-2 antigen by absorption of a specific antiserum, and the strength of the tumour-specific transplantation antigen by a transplantation assay, to see whether they are correlated . No obvious correlation was seen . We showed that cell suspensions of tumours taken directly from the animal are contaminated by host cells which make a substantial contribution in H-2 assays . Since this contamination was lost after several passages in vitro, the amount of H-2 on tumour cells was assayed only after such passages. J Cell Sci, 1978 Apr, 30, 319 - 30 PH oscillations in cell suspensions of Dictyostelium discoideum: their relation to cyclic-amp signals; Malchow D et al.; Cells of Dictyostelium discoideum known to release cyclic AMP (cAMP) rhythmically in the form of pulses, change with the same period of about 8 min the pH of their medium . The pH is used here as an indicator to investigate the effect of externally added cAMP pulses on the oscillations . Both a temporary increase in amplitude and a permanent phase shift can be induced . The phase-response curve indicates that the period can be increased and decreased by rhythmic stimulation with cAMP pulses. J Bacteriol, 1978 Apr, 134(1), 115 - 24 Oxygen-limited continuous culture and respiratory energy conservation in Escherichia coli; Rice CW et al.; Escherichia coli B was cultured continuously in succinate-minimal medium under conditions of oxygen limitation in the phauxostat . With decreasing oxygenation and consequent decreasing growth rates, the complement of terminal cytochrome oxidases changed as follows: high growth rates, cytochrome o; intermediate growth rates, cytochromes o and d; lowest growth rates, cytochromes o, d, and a1 . Respiratory kinetics exhibited by nongrowing cell suspensions obtained from continuous cultures indicated that terminal oxidase activity was exhibited by cytochrome o (Km for O2 = 0.2 micron; Vmax = 1.1 to 1.5 mumol of O2 per nmol of cytochrome o per min) and cytochrome d (Km for O2 = 0.024 micron; Vmax = 0.7 mumol of O2 per nmol of cytochrome d per min) . During oxygen-limited growth, the molar growth yield referred to respiration, and corrected for maintenance respiration {Yo(max)}, was 12.6 g (dry weight) per g-atom of oxygen, not significantly different from the succinate-limited value of 12.0 g (dry weight) per g-atom of oxygen . The rate of maintenance respiration of the oxygen-limited culture was only 3.4 mg-atoms of O per g (dry weight) per h, some threefold less than that of the succinate-limited culture . Respiration-driven proton extrusion did not vary with the growth rate or with the complement of terminal oxidases (H+/O = 3.7; standard deviation, 0.07) . We conclude that the content of terminal oxidases is without effect on the efficiency of respiratory energy conservation. Biochim Biophys Acta, 1978 Mar 30, 528(3), 456 - 65 Lipoprotein lipase of cultured mesenchymal rat heart cells . I . Synthesis, secretion and releasability by heparin; Chajek T et al.; Cell suspensions prepared from rat hearts were separated by replating into F1, F2 and M cultures, and cultured for 3--11 days . Lipoprotein lipase activity was highest in the F1 cultures which consisted mainly of non-beating, mesenchymal cells . The enzyme activity was released into the medium only after addition of heparin . The release occurred by an initial rapid phase and a continuous slow phase . Both the rapid and the slow release of enzyme activity by heparin were inhibited by about 70% after a 4 h pretreatment with colchicine . Thus, it seems that the vesicular transport is responsible for the translocation of lipoprotein lipase to the cell surface also during the slow process of release . The residual activity in the colchicine treated cultures was higher than in the controls indicating that no inhibition of enzyme synthesis occurred . The slow phase of enzyme release continued also after removal of heparin from the medium but was reduced markedly when protein synthesis was inhibited by cycloheximide . Thus the increase in total enzyme activity encountered after exposure to heparin resulted from stimulation of new enzyme synthesis . The half-time of lipoprotein lipase in the F1 cultures was 35 min and full restoration of enzyme activity was found 60 min after complete removal of cycloheximide from the system . These data indicate that the culture system can be used to study regulation of new enzyme synthesis and its turnover. Int J Cancer, 1978 Mar 15, 21(3), 329 - 33 Separation of lymphoid cells with a suppressor effect on the activity of cytotoxic cells in vitro during the growth of a syngeneic mouse tumour; Schaaf-Lafontaine N; One or 4 weeks after grafting of a syngeneic sarcoma (T2) to C57BL/6 mice, lymph-node cells (LNC) are cytotoxic in vitro for the cells of this tumour . But after 2 weeks LNC are not cytotoxic or show a non-significant activity . These second-week LNC, added to cytotoxic lymphocytes (CL) from the first or fourth week, reduce considerably the cytotoxicity of the latter cells . When velocity sedimentation at unit gravity is used to fractionate 11th-14th day LNC, some fractions, enriched in small lympoid cells, kill the cancer cells . Other fractions, containing large blast cells, lack this property but can suppress the activity of the small cells or of fourth week CL . These suppressor cells can also be separated from active CL by passage on a glass bead column . They are "adherent" while CL are non-adherent at this stage . This suppressor effect is abolished when the lymphoid cell suspension is treated by anti-theta serum, but macrophage depletion does not modify the inhibition . The cell responsible for the suppressor effect is thus a large T lymphoid cell, adherent and non-phagocytic . It seems to act essentially on an effector phase of the cell-mediated cytotoxicity. Eur J Biochem, 1978 Mar 15, 84(2), 487 - 92 Characterization of beta-glucosidase isoenzymes possibly involved in lignification from chick pea (Cicer arietinum L.) cell suspension cultures; Hosel W et al.; Crude cell wall preparations from Cicer arietinum L . cell suspension cultures show high activity for the hydrolysis of coniferyl alcohol beta-D-glucoside (coniferin) . Various beta-glucosidase activities could be solubilized from these preparations by 0.5 M NaCl treatment and one of these could be shown to possess a high activity for the hydrolysis of coniferin . The enzyme activities were purified to near homogeneity by Sephadex G-200 and CM-Sephadex chromatography . Isoelectric focussing indicated the occurrence of beta-glucosidase isoenzymes with identical catalytic activity (pI 8.5-10) . Molecular weights were determined as 110 000, with two subunits of 63 000 and 43 000 . Maximum hydrolytic activity is at pH 5 . The beta-glucosidase isoenzymes catalyze the hydrolysis of various beta-glucosides with aromatic aglycone structure and different sugar moieties . However, coniferin has been found to be one of the best substrates (km = 0.8 mM; V = 6 mumol.min-1.mg protein-1) for these beta-glucosidase isoenzymes . The data suggest that beta-glucosidase-catalyzed reaction might be involved in lignification of these plant cell cultures. Biull Eksp Biol Med, 1978 Mar, 85(3), 281 - 4 {Allogeneic hepatic cell suspension in the treatment of liver insufficiency}; Ostroverkhov GE et al.; A study was made of a possibility of using isolated hepatocytes for the treatment of diseases of the liver in animals . The optimal dose of the cell suspension in intravascular, intraperitoneal, intrapleural and subcutaneous administration was determined; a reaction of the experimental animals to this biological substrate was studied . The efficacy of the isolated hepatocytes in the treatment of hepatic insufficiency is demonstrated; also a comparative assessment of the mentioned methods of the cell suspension administration is given. Prostaglandins, 1978 Mar, 15(3), 437 - 46 The influence of some prostaglandins on DNA synthesis and DNA excision repair in mouse spleen cells in vitro; Egg D et al.; In vitro experiments were performed on mouse spleen cells to establish possible influences of some naturally occurring prostaglandins on DNA synthesis and DNA excision repair . The prostaglandins A1, B1, E1, E2 and F2alpha were tested in concentrations of lopg, 5ng and 2.5microgram per ml cell suspension . DNA synthesis was significantly increased by PgF2alpha in all the three concentrations tested, while the other tested prostaglandins were essentially ineffective . DNA excision repair was significantly inhibited by PgE1 and PgE2 at 5ng/ml and at 2.5microgram/ml but increased by PgF2alpha in the two lower concentrations . The rejoining of DNA-strand breaks after gamma-irradiation was slightly reduced by PgE1, PgE2 and PgF2alpha at 2.5microgram/ml. Mikrobiologiia, 1978 Mar-Apr, 47(2), 212 - 6 {Hydrogenase activity of Chlorella vulgaris cells}; Persanov VM et al.; Cell suspensions of Chlorella vulgaris were found to possess the hydrogenase activity as was confirmed by their ability to absorb H2 in the presence of benzyl viologen, azocarmine and other hydrogen acceptors as well as to produce H2 from reduced methyl viologen . Incubation of the cells in the dark under anaerobic conditions in the atmosphere of H2, N2 or Ar stimulated the activity of hydrogenase and induced its de novo synthesis . Treatment of the cells adapted to anaerobiosis with dry ice or liquid nitrogen considerably increased their hydrogenase activity . The enzyme of the adapted cells was more resistant to the inactivation by O2 and temperature. Phys Med Biol, 1978 Mar, 23(2), 324 - 31 Effect of ultrasonic irradiation on mammalian cells and chromosomes in vitro; Roseboro JA et al.; Human peripheral blood and HeLa cells were irradiated in vitro at the ultrasonic frequency of 65 kHz . The whole blood and HeLa cell suspensions were exposed to continuous and pulsed ultrasonic power levels of 0.12, 0.16, 0.72, 1.12 and 2.24 W for a period of one minute . The method of ultrasonic irradiation was carried out with the whole blood or HeLa cell suspensions coupled directly to a cylindrical transducer while heating of the cell suspensions in excess of 41 degrees c was avoided . Irradiated and unirradiated peripheral blood lymphocyte chromosome cultures were prepared and scored for selected numerical and morphological abberrations . There was no significant difference in the frequency of chromosomal aberrations between irradiated and unirradiated cells . The fraction of cells S surviving after an exposure to an ultrasound dose of D can be represented by S = exp (-D/Do) . Further, Do is shown to depend on the time after exposure at which survival is assayed. Invest Urol, 1978 Mar, 15(5), 380 - 4 In vivo growth of human bladder cancer cell lines; Heaney JA et al.; Two human bladder cancer cell lines grew predictably in rats immunosuppressed with antilymphocyte serum . Intraperitoneal inoculation of tumor cell suspensions resulted in diffuse intraabdominal carcinomatosis with consequent host death after 10 to 20 days . Subcutaneous inoculation of tumor cell suspensions resulted in local tumors which grew exponentially for 20 to 30 days before eventual regression after 40 to 50 days; lung metastases developed in at least 13 per cent of the animals with subcutaneous tumors . The histologic appearance of the xenografted tumors closely resembled that of the original tumors . Subsequent in vitro culture of the xenografted tumors provided cell lines that were morphologically identical to the primary lines and that retained a human karyotype . It is proposed to employ this model of human bladder cancer to evaluate chemotherapeutic agents for possible use in the clinical disease. Br J Surg, 1978 Mar, 65(3), 188 - 90 Cell viability studies on the exfoliated colonic cancer cell; Rosenberg IL et al.; Suspensions of desquamated colonic cancer cells were obtained from patients with cancer of the large bowel by colonic exfoliative cytology and from resected specimens of colonic cancer by an exfoliative technique . In addition, a tumor suspension was obtained from the resected specimens . Cell viability studies were performed on these cell suspensions . Whereas 23 of the 25 tumour homogenate cell suspensions were shown to exclude trypan blue, none of the exfoliated colonic cancer cell suspensions had viable cells . This finding would cast some doubt on the hypothesis that suture line recurrence following large bowel cancer surgery is due to the implantation of cells desquamated from the surface of the growth. Br J Cancer, 1978 Mar, 37(3), 337 - 44 Monocytes and macrophages in malignant melanoma . II . Lysis of antibody-coated human erythrocytes as an assay of monocyte function; Nyholm RE et al.; Peripheral blood mononuclear cells will lyse antibody-treated human erythrocytes . Using Group A red cells and a hyperimmune anti-A1 serum, we have devised a microassay for the cytolytic capacity of mononuclear cell suspensions . The effector cells responsible for red-cell lysis are mononuclear, adherent and phagocytic, and their activity is blocked by aggregated IgG . Their presence correlates well with non-specific esterase-containing cells and we conclude that they are monocytes . Dose-response curves of red-cell lysis plotted against numbers of monocytes were used to derive a simple parameter expressing the number of monocytes needed to lyse 15% of the 51Cr-labelled red cells . The assay was applied to a group of 27 normal controls and 36 patients with a histologically proven diagnosis of malignant melanoma . The results indicate that monocytes from patients show significantly greater lytic activity than those from the controls . These data suggest that monocytes from cancer patients are in some way activated, and that other defects in monocyte function which have been detected in cancer patients (defective chemotaxis and maturation) may be associated with monocyte "activation". J Reprod Fertil, 1978 Mar, 52(2), 387 - 90 Assessment by microflow fluorometry of the purity of interstitial cell suspensions from the rat testis; Clausen OP et al.; The technique of microflow fluorometry (MFF) was used to identify the proportion of haploid cells (from the tubules) in interstitial cell suspensions . The MFF estimates of the degree of contamination by tubular elements correlated well with the numbers of cells with Leydig cell morphology and those staining positively for 3beta-hydroxysteroid dehydrogenase. Br J Cancer, 1978 Mar, 37(3), 363 - 8 Arrest patterns of circulating lymphosarcoma cells in tumour-bearing mice as modified by previously injected cell suspensions; Weiss L et al.; The effects were determined of an initial i.v . injection of 0.2 ml suspensions of 125IUdR-labelled lymphosarcoma cells on the early arrest patterns of a second injection of cancer cells into tumour-bearing mice . The results indicate that interactions between the first injection and the host markedly affected the arrest pattern of the second dose in the lungs, but not the livers, of tumour-bearing animals . These observations are explained on the basis of the injected fluid volumes, which are considerable in mice, in relation to their total blood volumes of approximately 2 ml. J Lab Clin Med, 1978 Mar, 91(3), 444 - 54 Effects of phenobarbital and 3-methylcholanthrene pretreatment on size, sedimentation velocity, and mixed function oxygenase activity of rat hepatocytes; Sweeney GD et al.; SVA was used to separate liver cells from rats pretreated for 3 days with PB (40 mg/kg intraperitoneally, twice daily) or 3-MC (50 MG/KG AS A SINGLE INJECTIOn) . Twelve fractions of cells were collected with s's ranging from 97 mm/hr (fraction1) to 25 mm/hr (fraction 12) . Cells in each fraction were sized and counted electronically . PB caused the average volume of the largest cells recovered (fraction 1) to increase to 16, 725 micrometer3 from 10,500 micrometer3 previously reported in untreated animals . The number of cells recovered in the fast-sedimenting fractions (1 to 6) also increased, but there was only a small rise in the average model volume of hepatocytes determined prior to SVA . In cell suspensions analyzed after PB treatment no evidence was found for a discontinuity in the distribution of density-volume characteristics as previously described in hepatocytes from untreated rats . As expected, prior to sedimentation analysis, hepatocyte suspensions from rats treated with PB contained increased cytochrome P-450 . The average ratio (n = 4) of P-450 in separated to unseparated cells ranged from 2.75 (fractions 1 + 2) to 0.38 (fractions 11 + 12), giving a 6.8-fold range in quantity of cytochrome per cell . Over this range, cell volume increased 4.2-fold, indicating a gradient in P-450 concentration similar to that previously reported to exist in cells from untreated rat liver . The gradient for AHH activity was 6.7-fold, suggesting that activity of MFO's paralleled the increase in cytochrome concentration . 3-MC pretreatment caused no significant increase in either size or number of cells in the fast-sedimenting fractions, but the discontinuity in density-volume characteristics which distinguished fast and slow sedimenting cells of untreated rats became marked . Furthermore, both the size and number of cells recovered in fractions 7 to 11 (which include the modal peal volume of unseparated hepatocytes) were increased . The gradient of P-450 (OR P1-450) was less steep in 3-MC-treated cells with pooled fractions 1 and 2 containing an average of 4.62 times as much cytochrome as fractions 11 and 12 . The gradient for AHH was 4.29 times . The range of cell volume was 3.3-fold over this range . Additional experimental work was performed to determine whether P-450, AHH, or NADPH cytochrome c reductase exhibited differences in activity or concentration per unit of cell volume between cells on either side of fraction 7 where discontinuity had been noted . Each variable was expressed per unit of cell volume in fractions 5 and 9; ratios were compared but were indistinguishable from unity . It was concluded that induction of MFO activity had occurred equally in both populations of cells . Densities were calculated from s and cell volume . Noticeable loss of density followed PB treatment in cells of all sizes . 3-MC had less of an effect on density but enhanced the increment in density observed at fraction 7, which corresponds to the division between cells with distinct sedimentation characteristics... Clin Exp Immunol, 1978 Mar, 31(3), 524 - 5 B- and T-lymphocytes in toxic diffuse goitre; Skoldstam L et al.; Operation specimens of thyroid tissue from patients with Graves' disease were digested, with collagenase, down to a cell suspension . The lymphocytes in the suspension were concentrated by density-gradient sedimentation and the proportions of B and T lymphocytes were determined . Six patients with Graves' disease were studied . The proportions of B and T lymphocytes in the thyroid glands were similar to the proportions of B and T lymphocytes in the venous blood from healthy subjects. Endocrinology, 1978 Mar, 102(3), 937 - 46 Independence of steroidogenic capacity and luteinizing hormone receptor induction in developing granulosa cells; Hillier SG et al.; The relationship between FSH-induced acquisition of LH/hCG receptors and the steroidogenic capacity of granulosa cells from estrogen-primed hypophysectomized rat ovaries has been examined . Granulosa cells harvested from the immature preantral follicles of animals not treated with FSH (controls) displayed negligible specific human {125I}iodo-hCG binding and produced only minimal amounts of progesterone during 48 h of culture in vitro . Addition of highly purified hFSH or prostaglandin-E2 (PGE2) to the culture medium elicited substantial increases in progesterone production which were not accompanied by measurable increases in {125I}iodo-hCG binding . Treatment with oFSH in vivo for 24 h led to the initiation of antrum formation in many follicles and was accompanied by an 8-10-fold increase in hCG binding by freshly isolated granulosa cells . Basal, hFSH-, and PGE2-stimulated progesterone production during culture was also greater than controls . In contrast, cells from animals receiving oFSH in vivo for only 12 h showed no increase in hCG binding either before or after culture, yet basal and stimulated progesterone production in vitro was significantly greater than controls, indicating that the initiation of steroidogenesis was antecedent to LH/hCG receptor induction . Only those cells obtained after the 24-h in vivo treatment with oFSH produced elevated amounts of progesterone when incubated in the presence of hCG, thereby showing that the observed increases in {125I}iodo-hCG binding reflected the induction of functionally active LH/hCG receptors . Pharmacological stimulation of steroidogenesis by cell suspensions with N,O'-dibutyryl cAMP resulted in consistently high levels of progesterone production irrespective of previous treatment with FSH in vivo . This uniform expression of in vitro steroidogenic capacity occurred in the complete absence of measurable increases in LH/hCG receptors, suggesting that these two fundamental developmental processes are independent phenomena which may be under separate regulation in vivo. Eur J Immunol, 1978 Mar, 8(3), 180 - 4 Conditions for maximal synthesis of cyclic AMP by mouse macrophages in response to beta-adrenergic stimulation; Welscher HD et al.; The synthesis of cyclic AMP by mouse peritoneal macrophages in response to stimulation by isoproterenol was studied as a function of drug concentration, incubation time and cell density . Cyclic AMP levels of macrophages increased 3 to 3.5 times over the control level 20 sec after the addition of 10(-3) M isoproterenol . Under these conditions the dose response could be followed down to an isoproterenol concentration of 10(-6) to 10(-7) M . When cell suspensions were inactivated as early as 1 sec after the addition of the drug, the increase in cyclic AMP was much greater (153 vs . 25 pmol/10(7) cells) . Macrophage suspensions of high cell density were less responsive than those of low cell density . In the absence of any inhibitor of phosphodiesterase, the stimulatory effect of isoproterenol was always of short duration . The maximum effect of beta-adrenergic stimulation probably occurs in less than 1 sec at a cell density less than 2 x 10(6) cells/ml . The beta-blocking drug Visken abolished the observed effects. Anat Rec, 1978 Mar, 190(3), 687 - 702 Rapid isolation of chloride cells from pinfish gill; Hootman SR et al.; Enriched fractions of chloride cells with good ultrastructural integrity have been obtained from gill filaments of the euyhaline teleost, Lagodon rhomboides . The branchial epithelium from seawater-adapted fish was dissociated by gentle mechanical means in a Ca++, Mg++-free balanced salt solution . Density gradient centrifugation of the mixed cell suspensions through a Ficoll gradient yielded a fraction containing between 50 and 70% chloride cells . This fraction showed a 3- to 4-fold enrichment over comparable gill homogenate values for sodium plus potassium-activated adenosinetriphosphatase, (Na+, K+ ATPase), an enzyme concentrated in chloride cells . Isolation of chloride cells from fish adapted to one-third seawater was less successful, due to the smaller size and reduced number of these cells, although fractions with at least a 2-fold enrichment of the enzyme were obtained . These results continue to support the belief that chloride cells are responsible for osmoregulatory activity associated with the branchial epithelium of teleosts and that this vital function is mediated through the activity of the transport associated enzyme, Na+, K+-ATPase, the specific activity of which increases with osmotic stress. J Immunol, 1978 Mar, 120(3), 1074 - 6 Splenic suppressor cells and cell-mediated cytotoxicity in murine schistosomiasis; Coulis PA et al.; An impairment of the capacity to generate alloantigen-specific cytotoxic T lymphocytes (CTL) was observed in mixed lymphocyte cultures (MLC) established with spleen cells from mice infected with Schistosoma mansoni . This impairment, which was observed as early as the eighth week of infection, could be abrogated by the fractionation of spleen cell suspensions by the carbonyl iron/magnet method prior to the establishment of MLC . Cocultivation of normal spleen cells with increasing numbers of splenocytes from S . mansoni-infected syngeneic mice resulted in a dosage-dependent suppression of CTL generation . This "infectious suppression" was not sensitive to antiserum against mouse thymic lymphocyte antigen (MTLA) . The present studies suggest the role of a macrophage rather than a T cell as the suppressor cell in this model of cell-mediated immunity in schistosome-infected mice. Rev Fr Transfus Immunohematol, 1978 Mar, 21(2), 323 - 39 {Quantitative measurement of red cell antigens with Groupamatic}; Calvo A et al.; Groupamatic 360 can be adapted to the quantitative measurement of erythrocyte agglutination reactions . Here the authors give the necessary conditions to obtain the best sensitivity and the best reproductibility . They have considered globular suspensions, incubation of the erythrocyte antiserums mixtures, centrifugation, agitation, photometric measurement . As concerns the quantitative technique, we used the average peripheral measurement obtained in the twleve cuvettes of the same selected disc sector . The calibration is fixed to 50 mV for a non agglutinated red cell suspension and to 950 mV for albumin in a saline solution . In these conditions, the best discrimination is obtained for agglutination with measures between 400 and 800 mV . The maximum sensitivity is reached with reactions corresponding to 600 mV, that is to say in our selected conditions with 50% of free red cells . The choice of the zone between 500 and 600 mV enables to detect variations from 1.5 to 2% in an antibody solution concentration . This zone is particularly favorable for quantitative work . Although a part of these operations in this preliminary work were manually effected, the automatic centrifugation, agitation, photometric measurement and results print out make possible a large number of measurements with very satisfactory reproductibility conditions . Besides, these experiments pointed out some factors of improvement which will be helpful for the new 360 G Groupmatic equipments. Eur J Immunol, 1978 Mar, 8(3), 163 - 7 Subpopulations of mouse T lymphocytes . II . Suppression of graft-vs.-host reactions by naturally proliferating splenic T cells; Moorhead JW; The immunological role of a naturally proliferating subpopulation of splenic T cells was investigated using the graft-vs.-host (GvH) reaction on the mouse . Normal parental spleen cells, purified splenic T cells or lymph node cells were pulse-treated for one hour in vitro with tritiated thymidine of high specific activity ({3H}dThd, "thymidine suicide") . The treatment specifically and selectively kills proliferating cells which are actively synthesizing DNA, i.e . cells in S phase . Following treatment, the cells were transferred to F1 recipients and the GvH reaction measured by the splenomegaly assay . The results showed that the GvH effector cells in the donor spleen and lymph node are nonproliferating T cells . Furthermore, donor spleen cells treated with {3H}dThd consistently had enhanced GvH reactivity when compared to the controls, while the phytohemagglutinin response of these same treated cell suspensions was significantly inhibited . When purified splenic T cells were used, treatment with {3H}dThd also caused an increase in the GvH reaction, showing that a T cell population was being affected by the cycleactive agent . These results indicated that some naturally proliferating T cells have suppressor functions, and their specific inactivation allows nonproliferating effector T cells to mount a more vigourous GvH reaction. Acta Virol, 1978 Mar, 22(2), 104 - 12 Pyrimidine nucleotide synthesis in rat embryo cells infected with X14 or H-1 parvovirus; Ricceri G et al.; The activity of some enzymes involved in pyrimidine nucleotide synthesis was studied in rat embryo cell (REC) cultures infected with X14 or H-1 parvovirus . dCMP aminohydrolase activity of the infected cells was 64--121% greater than that of the mock-infected cells . dTMP synthetase activity was 18% greater in X14 virus-infected cells and 34% lower in H-1 virus-infected cells . These results suggest some differences in the infected cells, as regards the biosynthesis of dTMP . Orotate phosphoribosyltransferase and orotidylate decarboxylase activities appeared nearly unmodified compared to the mock-infected cells . The addition of phosphoribosylpyrophosphate (PRPP) to the cell suspension incubated with {6-14C} orotate increased the specific radioactivity of acid-soluble uracil, 5-fold in the mock-infected cells and 15- --24-fold in the X14 or H-1 virus-infected cells (72 hr p.i.) . This result suggests that the lowered pyrimidine nucleotide synthesis in infected cells depends to a large extent on the diminished PRPP pool. Thromb Haemost, 1978 Feb 28, 39(1), 146 - 57 Evaluation of platelet tests for measurement of cell integrity; Akkerman JW et al.; Various tests were evaluated for their capacity to differentiate between platelet suspensions with different degrees of cell damage . Those suspensions were prepared by simultaneous isolation of platelets from the same platelet-rich plasma (PRP) using the following procedures: 1 . centrifugation at 4 degrees C with EDTA 2 . gel filtration in Tangen's buffer 3 . gel filtration in Ca2+-free Tyrode's soltuion 4 . gel filtration in Ca2+-free Tyrode followed by dehydration against polyethylene glycol 20,000 and 5 . albumin density gradient centrifugation . In these suspensions and in the original PRP the following parameters were studied: 1 . morphology; 2 . aggregability upon ADP addition; 3 . platelet factor 3 availability; 4 . uptake of 14C-serotonin and 3H-adenine; 5 . metabolism of 3H-adenine and adenylate energy charge; 6 . endogenous total ATP, ADP and serotonin and 7 . lactate dehydrogenase (LDH) activity . Quantitation of pseudopod formation in the light or electron microscope and log dose response studies for ADP-induced aggregation proved to be the most sensitive and reproducible of the tests studied . Additional information could be obtained from measurement of the 3H-label in the ATP and hypoxanthine-inosine fractions and calculation of the adenylate energy charge . Determination of platelet factor 3 availability or uptake studies of 14C-serotonin and 3H-adenine were less suitable for discriminating between cell suspensions . Data for total ATP and serotonin concentrations and LDH activity differed between the cell suspensions but instead of detecting various degrees of cell damage they reflected alterations in platelet population caused by the isolation procedures. Biomedicine, 1978 Feb, 29(1), 22 - 5 Preparation of single cell suspension from rat liver . A simple method designed for immunologic studies; Rachman F et al.; Single cell suspensions from rat liver were prepared by mean of a 10 min . in vivo liver perfusion with an isotonic solution . This solution was devoid of calcium but contained 27 mM of sodium citrate and 1% of bovine serum albumin . The liver cells were dissociated over chromium nickel screen in presence of 0.8 mM of calcium and 0.25 M of saccharose; they were further separated by gravity sedimentation for 10 min . The dissociated cells, when incubated in Waymouth's medium enriched with 15% of heat-inactivated horse serum and 1% of BSA, actively synthetized urea and proteins . They also incorporated {3H} thymidine . Isolated hepatocytes, obtained without resorting to extraneous enzymes, conform to the conditions required for the study of cell surface receptors involved in immunologic recognition mechanisms. J Lipid Res, 1978 Feb, 19(2), 269 - 73 Determinations of adipose cell size and number in suspensions of isolated rat and human adipose cells; Cushman SW et al.; The osmic acid fixation-Coulter electronic counter method described for determining adipose cell size and number in intact adipose tissue fragments has been modified for use with suspensions of isolated rat and human adipose cells . Mean cell sizes in tissue fragments and isolated cell suspensions prepared from the same tissue are virtually identical in rats of various weights . No statistically significant difference in mean adipose cell size between tissue and isolated cell suspension was observed in human adipose tissue although the variability was much greater than in rat tissue . The distribution of cell sizes among replicate samples is more uniform in the isolated cell preparations, possibly reflecting the considerably larger quantities of tissue used in preparing isolated cells than in determining cell size and number directly from tissue fragments . An example of the utility of the modified method during routine metabolic studies with isolated rat epididymal adipose cells is described; isolated cells of increasing size can be obtained from rats of increasing body weight, or from the separated distal and proximal portions of the fat pads of rats of the same weight. Biull Eksp Biol Med, 1978 Feb, 85(2), 219 - 21 {Effect of regeneration of hematopoietic organs on the number and type of splenic colonies}; Sukhova GK et al.; Two thirds of the spleen or the bone marrow of the tibia were removed in CBA mice . Hemopoietic splenic colonies were obtained and examined microscopically on the 8th day after the lethal irradiation of mice and injection of the spleen cell suspension . An increase of the number of colonies and a significant rise of the number of granulocytopoietic colonices in comparison with their number in control was seen in the experimental animals . The number of other hemopoietic colonies remained unchanged . The authors suggest that these changes could be caused by the local influence of the proliferating spleen stroma, or by a factor secreted from the regenerating stroma of the hemopoietic organs. Br J Cancer, 1978 Feb, 37(2), 248 - 53 Quantitative relationship between volume of tumour cell units and their intravascular survival; Lione A et al.; The derivation of the median volume (MV) and the geometric standard deviation (SDg) for a suspension of tumour cells quantifies the size and distribution of tumour cell aggregates in the suspension . Data collected in a group of 14 experiments shwoed a significant correlation of 0.80 (P less than 0.001) between the number of lung tumours formed by a suspension of B16 melanoma cells injected i.v . into C57BL/6J mice and the product of the MV and SDg of each cell suspension . These data define a size parameter of tumour cell suspensions that correlates with the intravascular survival properties of tumour cells. Acta Pathol Microbiol Scand {C}, 1978 Feb, 86(1), 11 - 7 Functional analysis of lymphoid cells from mouse bone marrow in vitro; Rubin B et al.; The present results show that mouse bone marrow (BM) cells responded in vitro against phytohaemagglutinin (PHA), concanavalin A (Con A) and lipopolysaccharide (LPS) . In addition, BM cell suspensions contained relatively many cells with ADCC (antibody-dependent cell-mediated cytotoxicity) activity . BM cells were fractionated according to various cell surface characteristics . The results of these experiments were compared with previously published reports, showing that pre-T and pre-B cells could be separated on Ig anti-Ig columns (Ig = immunoglobulin) . The combined results showed that pre-T cells may fractionate together with cells with the following characteristics: they displayed ADCC activity and had Fc receptors (FcR) and/or membrane Ig . Pre-B cells fractionated with cells with the following characteristics: they responded to LPS but had no ADCC activity . They were not adherent and/or phagocytic and were Ig-, FcR- and C3 receptor (C3R) negative. Cytobios, 1978, 23(91-92), 199 - 23 Reconstruction of mammalian heart tissue from embryonic heart cell suspension with reference to the aggregation of adult heart cells; Nag AC; Aggregations of isolated embryonic and adult heart cells were studied so as to examine in detail the formation of cell contacts, the assembly of cells into multicellular systems, and cell co-operation in forming organized and differentiating tissues . At selected intervals after initiating rotation cultures, aggregates were examined microscopically for evidence of contractility, and subsequently processed for scanning and transmission electron microscopy (TEM, SEM) . By continuous accretion of single cells, and the joining of small clusters, the aggregates increased in size . Cells within the aggregates exhibited rhythmic and synchronous contractility by 3--12 h of culture, suggesting the formation of low-resistance inter-cellular junctions between apposed cells . Two populations of cells could be recognized by 9--12 h with SEM . One was spherical in surface view and the other was flattened . Spherical cells possessed myofibrils, and were classified as cardiac myocytes which occupied the core of the aggregate . The flattened cells were devoid of myofibrils and non-muscle in nature . They covered the surface in a multilayered epithelium . At 12h the aggregates were round to oval, covered by flattened cells, but individual round cells could still be recognized . Intercalated discs were frequently observed in 12 h aggregates . The junctional complexes observed in 12--72 h aggregates include desmosomes, fascia adherens and gap junctions . Most of the aggregation was completed by 24 h, and at later time periods, i.e . 48--72 h, the external surface of the aggregates was smoothed out with epithelial investment . In these aggregates myofibrils and intercellular junctions became reconstructed in less than 24 h . Unlike embryonic myocardial cells, adult cells did not form aggregates of numerous cells . Instead, they formed irregular clusters of 2--5 cells during 3--48 h of culture . Intercellular contacts and suggestive desmosomal materials were observed between adherent cardiac muscle cells . When culture continued for 48--72 h, the cells underwent supercontraction and became non-viable, suggesting that the terminally differentiated adult myocardial cells are incapable of regenerating constituents obligatory for histogenetic reconstruction. Arch Immunol Ther Exp (Warsz), 1978, 26(1-6), 89 - 93 Studies on cell surface antigens of mouse leukemic and normal lymphocytes . I . Categories of cell surface antigens on mouse leukemia and their serological identification; Radzikowski C; Analysis of cell surface antigens is most advanced with mouse tumors, mainly because of the availability of inbred strains with known susceptibility to naturally occuring or induced tumors . The development of serological techniques enables identification of gene products which are expressed on the surface of tumor cells . The naturally occuring and induced leukemias in mice are a particularly suitable model for the studies of surface antigens, because the leukemia cells can be easily obtained in cell suspension, they are highly sensitive to cytotoxic antibodies and they can be compared, because of common origin, with normal thymocytes and lymphocytes . In addition to conventional alloantigens (MHC), differentiation alloantigens (Thy, Tl and Lyt) viral structural (MuLV and occasionally MMTV) also viral related cellular antigens are detectable . Various categories of cell surface antigens and antisera defining their presence on the surface of mouse lymphocyte produced and used for studies carried on in the Department of Tumor Immunology are listed and discussed in the present and following papers (No . I-V). J Nutr Sci Vitaminol (Tokyo), 1978, 24(4), 383 - 95 Studies on the factors influencing the hydrogen peroxide hemolysis test; Mino M et al.; There seems to be a greater variation between laboratories in hydrogen peroxide hemolysis techniques than in other clinical laboratory procedures . In this report, factors influencing hemolytic values were examined and the effect of the combination of these factors on the results was analysed statistically by using the orthogonal array . Factors influencing hemolysis induced by hydrogen peroxide were as follows; the concentration of hydrogen peroxide, temperature in the peroxide reagent when added to the red cell suspension, the red cell concentration in the cell suspension and the addition of charcoal to the reaction mixture . However, addition of charcoal may not be essential to stabilize the hemolytic values and other factors such as keeping blood for 4 hours at room temperature before testing, the difference between investigators, a reaction time 2 or 3 hours and the technique of adding peroxide reagent to the cell suspension, had little effect on hemolysis . The most important factor was the temperature in hydrogen peroxide solution . The estimated hemolytic values by the orthogonal array linearly correlated with the plasma tocopherol levels. Arch Geschwulstforsch, 1978, 48(2), 145 - 9 Pulse cytophotometric measurements on tumor cell suspensions and exfoliative material; Krug H; By impulse cytophotometry (ICP) it is possible to get a DNA-distribution curve of nuclei in cell suspensions . The ICP curve directly represents the ploidy pattern of the investigated cell population and, more indirectly the proliferation intensity . For practical tumor diagnosis in cytology by ICP as a first step we must know, wether the ICP is able to detect differences between normal and tumor cells by simple criteria . Together with our clinical coworkers we had investigated cell suspensions from carcinomas of the cervix, ovary, breast, larynx, stomach and malignant melanomas . Approximately one half of these tumors were poly- or aneuploid . In the remaining diploid tumors some did not show signs of strong proliferation in the form of high 4c-peaks in the ICP; for these the chances of detecting by ICP in exfoliative material are low . This is in agreement with our results on vaginal smears . Here we had a good correlation between the mean 4c-height of the ICP curves and the Papanicolaou grading . But mass screening is not the main application of ICP because the security of Pa-grading is more sufficient . In individual problematic cytological cases with repeated investigation ICP is a very valuable diagnostic aid. Cytogenet Cell Genet, 1978, 21(5), 267 - 81 The chromosomes of cockerels (Gallus domesticus) during meiosis; Pollock DL et al.; The chromosomes in spermatogenic cells of 10 cockerels were examined . Prior to removing the testes, the birds had been injected with either 2.5 mg vinblastine sulfate or 5 mg colchicine, or were untreated . A cell suspension was made from small pieces of testis and slides were prepared using an air-drying technique . Pretreatments of the birds increased the numbers of spermatogonial and secondary spermatocyte cells at metaphase and rendered them more amenable to analysis . The modal number of macrochromosomes or bivalents was observed in 90.2% of spermatogonia, 98% of primary spermatocytes at diakinesis, and 94.2% of secondary spermatocytes . Polyploid cells at all stages were thought to be largely artifacts of the procedure . Aneuploidy was rare, being found in only 1% of secondary spermatocytes . It is estimated that between 56 and 66 chiasmata occur in the 39 bivalents; of these, an average of 28 are seen in the six macrobivalents . Average numbers were 8.1, 5.9, 4.6, 3.6, 2.4, and 3.5 for macrobivalents 1 through 5 and the Z respectively . The numbers of chiasmata differed significantly among birds but were not differentially affected by pretreatment of the birds. J Immunol Methods, 1978, 21(1-2), 79 - 88 Classical and alloimmune anaphylactic degranulation of isolated single mast cells; Thiernesse N et al.; Viable mast cells, directly isolated by micromanipulation from a mouse peritoneal cell suspension, were deposited on the bottom of microtiter-plate wells and submitted to histamine release . Conventional antigen-induced anaphylactic degranulation as well as direct allogeneic anaphylactic degranulation were strongly inhibited when these mast cells were settled on normal tissue culture plastic surfaces . Nevertheless, normal degranulation could be recovered by pretreatment of the experimental surface with a multipositive charged molecule (poly-L-lysine) . Under these conditions, we demonstrate that the degranulation of one isolated mast cell is possible and consequently, as regards the direct allogeneic anaphylactic degranulation, confirm the "self-triggering mechanism" in which the recognition of histocompatibility antigens on the membrane of the mast cell itself is the trigger to the secretory response . The technique of monocellular degranulation described in this paper provides a new tool which leads us to think that the problem of detection of anaphylactic antibody-secreting cells can be solved. J Immunol Methods, 1978, 20, 173 - 83 A sensitive rosetting method for detecting subpopulations of lymphocytes which react with alloantisera; Parish CR et al.; A procedure is described for directly estimating the proportion of mouse lymphoid cell suspensions which react with alloantisera . The method entails reacting Ig-capped lymphoid cells with alloantisera, and then assessing the uptake of alloantibodies by rosetting the lymphocytes with SRBC coated with sheep IgG specific for mouse Ig . This rosetting procedure was found to be generally more sensitive than the conventional dye exclusion microcytotoxicity test for detecting the binding of alloantibodies to lymphocytes . Furthermore, the rosette method has the advantage that, unlike the complement lysis technique, it has a low and reproducible background and lymphocyte subpopulations which react with alloantisera can be isolated. Cytogenet Cell Genet, 1978, 20(1-6), 365 - 72 Studies on the function of H-Y antigen: dissociation and reorganization experiments on rat gonadal tissue; Zenzes MT et al.; On circumstantial evidence, H-Y antigen is assumed to be responsible for the differentiation of the undetermined gonadal anlage into a testis . A direct approach to test the function of H-Y antigen is provided by Moscona-type experiments . Applying a modified technique of in vitro reassociation of cell suspensions, we obtained the following results: (1) dissociated newborn rat gonads, both testis and ovary, reorganize into histotypic structures; (2) under exposure to anti-H-Y antiserum, testicular cells reassociate into ovarian follicular-like organization; (3) anti-H-1 antiserum by itself does not prevent the testicular cells from forming tubular structures . It is concluded that H-Y antigen acts as a differentiation between preferably or exclusively on the cell elements participating in the formation of the seminiferous cords. Biochimie, 1978, 60(1), 35 - 44 Glycosyl transfers from UDP-sugars to lipids of plant membranes: identification and specificity of the transferases; Axelos M et al.; Microsome preparations extracted from wheat roots or sycamore cell suspensions catalyzed the transfer of sugar from nucleotide-sugars to endogenous lipidic acceptors . The nature of the products biosynthesized from UDP-Glc, GDP-Glc, UDP-Gal, UDP-Xyl or UDP-Arab was examined . Sterylglycosides were obtained from UDP-Gglc, GDP-Glc or UDP-Xyl . Galactosyldiglycerides were synthesized from UDP-Gal . When UDP-Glc or UDP-Gal was used as a substrate, a membrane-bound 4-epimerase interconverted the epimeric nucleotide-sugars, thereby allowing the simultaneous biosynthesis of galactosyldiglycerides and sterylglucosides . The biosynthesis of free and acylated sterylglucosides from UDP-Glc, without interference of other glycosyl transfer reactions, was obtained by the omission of Mg++ ions from the incubation medium . The biosynthesis of galactosyldiglycerides from UDP-Gal without interference of other transfer reactions was obtained when digitonin was added to the incubation medium of sycamore microsomes. Hemoglobin, 1978, 2(2), 101 - 16 Acylation of hemoglobin by glutarylsalicylamide and its effect on oxygen transport properties; Tam JW; Hemoglobin A was modified in vitro with 0.02-0.03 M glutarylsalicylamide for two hours at pH 7.2 and 37 degrees C . The extent of modification was about 30-50%, as estimated by visual comparison after electrophoretic separation . A substantial decrease in oxygen affinity of modified hemoglobin solutions was observed . Similar results were also obtained for dilute cell suspensions of washed red blood cells and whole blood after GSM modification . Other properties such as cooperativity, Bohr effect and 2,3-DPG dependence remained essentially unchanged . Athough the site(s) of modification have not been determined, it is unlikely that they would involve any amino acid residue contributing to the above allosteric properties. J Natl Cancer Inst, 1978 Jan, 60(1), 179 - 83 Isolation from a murine fibrosarcoma of cell lines with enhanced plating efficiency in vitro; Suzuki N et al.; The technique for the isolation of mutants was applied to establish highly clonogenic cells from a fibrosarcoma (FSA) that had an extremely poor growth capacity in vitro (10(-6)--10(-7) of surviving fraction of cells) . After consecutive clonings, the surviving fraction of cells increased to 1--5 X 10(-1), whereas that of the ordinarily maintained culture remained at a low level . Selected clones were analyzed in vitro and/or in vivo . The results indicated that the FSA was composed of heterogeneous cells or cells having a potential variability in cloning ability in vitro, metastatic ability in vivo {lung colony-forming efficiency (LCFE)}, and DNA content . The relatively high DNA content of one of the clones, FSA 1233, continued after growth in vivo or in vitro, indicating its hereditary nature . FSA 1233 was also demonstrated to have a lower LCFE when the cell suspension was made from a tumor rather than from a culture in vitro . The difference of surviving fractions between the cells in the two conditions was not enough to explain the difference in LCFE's between them . These cells could grow under either in vitro or in vi