J Cell Physiol, 1982 Apr, 111(1), 97 - 103 Characterization of a postlavage, in situ pulmonary macrophage population; Drath DB et al.; A postlavage in situ subpopulation of pulmonary macrophages (PM), biochemically distinct from the lavaged population, has recently been isolated from rats . After exhaustive bronchopulmonary lavage to extract the free lung cells, the lungs were excised, homogenized, and filtered, and the resultant cell suspension was allowed to form a monolayer on plastic Petri dishes . Electron microscopic morphometry failed to indicate any morphologic differences in the two populations . The postlavage in situ PM were more active metabolically during phagocytosis of zymosan particles or stimulation by phorbol myristate acetate (PMA) than the corresponding lavage population, as evidenced by greater superoxide generation . Macrophages prepared by either method became more avidly phagocytic when incubated with cell-free medium isolated in the preparation of the situ population . Peroxidase, an enzyme absent from the granules of PM separated by lavage techniques, was found in a granule-rich fraction of the in situ macrophage . Catalase activity was found in similar amounts in both supernatants and granule-rich fractions of both populations . The results support the concept of subpopulations of PM and suggest that these subpopulations are distinguished by their biochemical properties and their functional abilities. J Clin Endocrinol Metab, 1982 Apr, 54(4), 689 - 92 The effect of prolactin on human aldosterone-producing adenomas in vitro; Carroll JE et al.; There is evidence for an unidentified aldosterone-stimulating factor of pituitary origin . We measured the effect of ovine PRL (oPRL) on aldosterone secretion by isolated cell suspensions of human aldosterone-producing adenomas (APAs) and compared it to the effects of angiotensins, ACTH, and potassium (K+) . In the first APA, the aldosteronotropic action of large doses of oPRL was double that of angiotensin II (AII); the response to ACTH was triple that to AII, while K+ had a small stimulatory effect . Results with the second APA showed that physiological concentrations of oPRL caused a response nearly double that to AII, but, once again, less than the response to ACTH; K+ was inert . ACTH contamination of the oPRL preparation was too minute to account for these findings . We conclude that oPRL possesses aldosterone-stimulating activity in APAs greater than that of angiotensins and potassium, but less that that of ACTH . These data suggest a role for PRL in aldosterone secretion by aldosterone-producing adenomas. Agents Actions, 1982 Apr, 12(1-2), 60 - 3 Histamine release from human pulmonary mast cells; Ennis M; The enzyme collagenase was used to disperse human lung into its component cells . The resulting cell suspensions contained circa 8% mast cells and were used for studies of mediator release without further purification . They exhibited a low (circa 7%) spontaneous release of histamine . They could be sensitized passively and released histamine upon challenge with anti-human IgE . They responded to concanavalin A but not to dextran . Phosphatidyl serine did not potentiate the release induced by these agents . The calcium ionophores, A23187 and ionomycin, both elicited histamine release . The cells were refractory to the action of the basic releasers 48/80 and peptide 401 (MCD-peptide) . These results indicate marked differences between human pulmonary mast cells and the more widely used rat peritoneal mast cells. Nouv Presse Med, 1982 Mar 27, 11(14), 1071 - 4 {Clonogenic cultures of human tumours . Experience and prospects in cancer chemotherapy (author's transl)}; Rozencweig M et al.; Malignant cells grow selectively in semi-solid media . Cultures of single cell suspensions in these media result in the formation of colonies which are believed to represent clones of stem cells . Cells freshly sampled in cancer patients are assayed in such a system to quantitate the in vitro effect of various anticancer agents vs untreated controls . Partially retrospective comparisons between in vitro and in vivo data are very encouraging and suggest that observations in this assay correlate very favorably with clinical findings . This clonogenic assay should provide an experimental model of special relevance to clinical cancer chemotherapy and should greatly help define optimal treatments in individual patients . Although many technical problems remain to be settled, prospective studies are ongoing to determine to what extent the assay is actually predictive of the response to chemotherapy in cancer patients. Int J Radiat Oncol Biol Phys, 1982 Mar-Apr, 8(3-4), 737 - 9 Binding of misonidazole to EMt6 and V79 spheroids; Franko AJ et al.; Metabolism-induced binding of misonidazole was shown previously to be enhanced in hypoxic regions of V79 spheroids . Data are presented to show that the binding of misonidazole to EMT6 spheroids is similar to V79 in some respects and different in others . The binding rate in nitrogen was constant in all cells in EMT6 spheroids, whereas a 5-fold rise was found in binding rate across the outer 100 micrometers of V79 spheroids . A small enhancement of binding to the cells near the necrotic center in EMT6 spheroids in air was found, which was similar to the binding to chronically hypoxic cells in aerobic V79 spheroids . The binding rate in 1--5% oxygen was similar in EMT6 and V79 spheroids . Both showed a large rise in binding rate across the outer 120 micrometers and a constant binding rate to healthy cells interior to this . Since 1.5 oxygen should supply only the outer 40 micrometers of a typical spheroid, something beyond a simple oxygen diffusion limitation is required to explain the data . One possible explanation is the diffusion of some of the reactive product away from the hypoxic cells which produce it . This was tested by incubating a single cell suspension in misonidazole at 0.5% oxygen in the presence of absence of V79 spheroids . No evidence was found for an appreciable loss of reactive product from the spheroids. J Pharmacol Methods, 1982 Mar, 7(2), 153 - 9 A simple method for preparing single cell suspensions of heart and smooth muscle for radioreceptor labeling studies; Johns A et al.; A simple method of preparing single cell suspensions for radioligand binding studies is described . The method involves incubating tissues in the presence of collagenase and elastase for 90 min in physiological solution with 1 mM calcium chloride and the mechanical disruption of the tissue by pipetting . The tissues examined were atria, ventricle, bladder, uterus, and taenia coli removed from mature guinea pigs . Viability of the cells by trypan blue exclusion showed 60-88% viable cells and receptor binding studies using (3H}-1-quinuclidinyl benzilate (QNB) yield KD values of approximately equal to 0.1 nM . The receptor numbers for each tissue were (receptors/cell): atria, 5700; ventricle, 11,000; uterus, 31,000; bladder, 44,000; taenia coli, 68,000. Steroids, 1982 Mar, 39(3), 245 - 58 The specific binding of estradiol and estrone and the subsequent distribution of estrogen-receptor complexes within MCF-7 human breast cancer cells; MacIndoe JH et al.; We have examined the specific binding of estradiol (E2) and estrone (E1) within MCF-7 cells using both cytosol and whole cell suspension binding assay techniques . The results of these studies have revealed that each of these estrogens binds the high but different affinity to the same single class of cytosol receptor (ER) . The incubation of intact cells with E2 at 37 degrees resulted in the rapid formation, nuclear binding, and nuclear processing of E2-ER complexes . Reduction of incubation temperature to 15 degrees C and 4 degrees C resulted in slowed but continued E2-ER formation and nuclear binding, and in a marked inhibition of nuclear receptor processing . The incubation of intact cells with E1 at 37 degrees also was associated with the formation, nuclear binding, and nuclear processing of estrogen-ER complexes . Interestingly, however, E2 rather the E1 represented the major species of specifically bound estrogen under these conditions . This observation suggests that the estrogenic action of E1 in MCF-7 cells may be mediated largely by the intracellular formation of E2 . This phenomenon provides a likely explanation of the unexpected potency of E1 in this system previously observed by other workers . Furthermore, our results suggest that E1 may represent a major source of estrogenic stimulation for some hormone-dependent human breast tumors. Res Vet Sci, 1982 Mar, 32(2), 253 - 6 Microtitre techniques for the assay of rinderpest virus and neutralising antibody; Rossiter PB et al.; Microtitre techniques were compared with conventional tube techniques for their ability to assay rinderpest virus and neutralising antibody to the virus . The microtitre technique was as sensitive and reliable for assaying the virus as the recommended tube technique, using cell suspensions . Both of these methods, however, were less sensitive than tube titrations on preformed cell monolayers . The microtitre test was as sensitive as the tube test for detecting and assaying virus neutralising antibody and more robust in that it was less sensitive to variations in virus dose. J Immunol, 1982 Mar, 128(3), 1070 - 5 Studies on natural killer (NK) cells . III . The effects of in vitro culture on spontaneous cytotoxicity of murine spleen cells; Bartlett SP et al.; In vitro culture of normal lymphohemopoietic cells is a complex event leading to the generation of a diverse group of effector cells . When murine spleen cells are cultured by themselves in fetal calf serum containing medium for up to 9 days, at least 3 distinct subpopulations of cytotoxic effectors can be distinguished on the basis of tumor target cell preference and expression of cell surface alloantigens . Fresh spleen cell suspensions contain 2 distinct NK cell types: NK-1.2+ NKA cells, which preferentially lyse lymphoma targets, and NK-1.2- NKB cells, which lyse nonlymphoma, mainly solid tumor targets . These NK cells account for all the cytolytic activity on day 0 of culture . NK-1.2+ NKA cells are very labile, and greater than or equal to 80% of their activity is lost within 48 hr of culture . By contrast, the activity of NKB cells increases in culture, up to 4-fold by day 6, and solid tumor target cells resistant to lysis by fresh spleen cells now become susceptible . In addition, an NK-1.2-, H-2+, Thy-1.2+, Ly-1.2+, Ly-2.2+ cytotoxic effector cell arises in culture from, or is dependent upon the presence of, NK-1.2-, Thy-1.2+ cells . This spontaneously arising "Tc" has a peak of activity between day 3 and 6 of culture, and kills a variety of lymphohemopoietic tumor cell types. Biull Eksp Biol Med, 1982 Mar, 93(3), 63 - 5 {Ultrastructural localization of type-specific antigen of legionella pneumophila}; Popov VL et al.; Cell suspensions of Legionella pneumophila, a virulent Philadelphia I strain, were incubated with rabbit antiserum to type-specific heat-sensitive antigen III having toxic properties with the use of the indirect immunoferritin technique . Antigen III was localized in the outer fibrillar microcapsule-like layer 12-20 mm thick . In the direct immunoperoxidase test antigen III had globular localization on the cell wall surface . The layer described is unlikely to be a true microcapsule, since it does not contain acid mucopolysaccharides and is readily removed from the cell . It is more likely that it forms as a result of antigen III secretion (and, possibly, of other antigens) being more similar to a capsule-like or mucilagenous layer. Biochimie, 1982 Mar, 64(3), 185 - 93 In vivo study of cholesterol turnover in tissues of adult sows; Aigueperse J et al.; The in vivo study of free and esterified cholesterol turnover was carried out in 15 tissues of adult Large White sows maintained at a constant weight for 10-12 weeks . They received a single intravenous injection either of {1-14C} acetate, or of an autologous red cell suspension or of plasma, previously labelled in vitro (for red cells) or in vivo (for plasma) with tritiated cholesterol . The tissues can be separated into four groups according to their relative rate of free cholesterol exchange between plasma and tissues . The liver and the lungs have a very fast exchange rate whereas the brain and the spinal cord have a very slow one . The whole lipoprotein particle transfer--an exclusive model for the esterified cholesterol transport from plasma to tissues--has been found in all sow tissues . When {1-14C} acetate is used as a substrate for cholesterol synthesis, lungs, adrenal glands and heart do not seem--or at an extremely low rate--to convert acetate into cholesterol whereas an intense cholesterol synthesis takes place in the small intestine . Its contribution to cholesterol synthesis in sows--taking into account the cholesterol transfer processes--reaches 70 per cent. J Biol Chem, 1982 Feb 10, 257(3), 1128 - 30 Inactivation of yeast fructose-1,6-bisphosphatase . In vivo phosphorylation of the enzyme; Mazon MJ et al.; Incorporation of 32P into yeast fructose-1,6-bisphosphatase (EC 3.1.3.11) was observed after addition of glucose to a cell suspension incubated with (32P)orthophosphoric acid . The 32P counts were coincident with the enzyme band when immunoprecipitates were subjected to sodium dodecyl sulfate disc gel electrophoresis . The incorporation of phosphate was associated with a decrease in enzyme activity . Approximately 1 mol of phosphate was incorporated/mol of enzyme . The phosphate is bound to the enzyme in a phosphoester linkage with a serine residue . Release of 32P accompanying enzyme reactivation was observed both in vivo and in cell-free extracts. Tissue Antigens, 1982 Feb, 19(2), 134 - 9 A method for freezing sheep lymphocytes prior to cytotoxicity testing; Stear MJ et al.; The two stage freezing technique was adapted for use with sheep lymphocytes . Parameters investigated were lymphocyte concentration, composition of thawing medium and DMSO concentration . The main difference between this and earlier published techniques is the use of a high DMSO concentration (17%) . The technique adopted was both simple and reliable . It consistently gave lymphocyte viabilities of 95% or more in thawed cell suspensions . The procedure is apparently without effect on lymphocyte antigen expression and thus appears very suitable for use in microlymphocytotoxicity tests. J Steroid Biochem, 1982 Feb, 16(2), 175 - 84 Receptor mediated gonadotropin action in gonadal tissues: relationship between blood cholesterol levels and gonadotropin stimulated steroidogenesis in isolated rat Leydig and luteal cells; Azhar S et al.; The present studies were performed to evaluate the role of steroid precursors and plasma lipoproteins in gonadal tissue steroidogenesis . Leydig cell suspension isolated from rat testes responded to hCG . Bt2cAMP, 8 Br-cAMP and cholera toxin with an increase in testosterone response . Administration of 4-aminopyrazolo{3,4-d}pyrimidine (4-APP) reduced the plasma cholesterol and testosterone levels in a time and dose dependent manner . This treatment also reduced the steroidogenic capacity of isolated Leydig cells both under basal conditions and in response to trophic hormone . Different doses of 4-APP up to 25 mg/kg BW and up to 4 days of treatment, however, did not modulate cholesterol and cholesterol ester contents of isolated Leydig cells . 4-APP treatment also had no effect on testis weight, phospholipid content, protein synthesis and energy metabolism in isolated Leydig cells . Similarly, administration of 4-APP (12.5 mg/kg) to PMSG-hCG primed rats beginning on day 3, post hCG, drastically reduced the circulating cholesterol and progesterone levels . Injection of the drug also produced an inhibition in vitro luteal cell steroidogenesis and a reduction in cellular cholesterol esters and free cholesterol contents . Addition of LDL or HDL to incubation medium reversed the inhibitory effect of 4-APP on luteal cell steroidogenesis while this inhibition persisted in Leydig cells . Injection of rats with Triton-WR-1339 (mg/kg BW) resulted in a 10-fold increase in plasma cholesterol and a contrasting decrease in testosterone levels . This treatment, however, produced no effect on in vitro Leydig cell steroidogenesis or cellular content of cholesterol esters and free cholesterol . It appears that the Leydig and luteal cells process and utilize lipoprotein-delivered cholesterol for steroidogenesis through different mechanism(s) . These studies thus demonstrate differential actions and an acute regulatory role of lipoproteins in gonadotropin modulated steroidogenesis in two different gonadal tissue. J Toxicol Environ Health, 1982 Feb, 9(2), 277 - 85 Comparison of methods of assessment of metal-induced lipid peroxidation in isolated rat hepatocytes; Stacey NH et al.; There is some controversy about which method of assessment of lipid peroxidation in isolated hepatocytes is most appropriate . The present study was undertaken primarily to compare measurement of concentrations of thiobarbituric acid (TBA) reacting substances with measurement of ethane concentrations in the gas phase of the incubation flask as indicators of lipid peroxidation . Four metal salts, FeCl2, NaVO3, CdCl2, and MnCl2, were selected as agents that interact with the lipid peroxidation process . Furthermore, reduced glutathione (GSH) concentrations and enzyme leakage were assayed to determine whether there was a consistent pattern of interaction between lipid peroxidation and change in GSH concentrations and enzyme leakage . The effects of the metal ions on the concentration of TBA reactants estimated in the whole cell suspension and on the gaseous ethane concentration were similar . However, the assessment of TBA reactants was a little more sensitive, and ethane concentrations continued to climb with incubation time rather than leveling off as observed for TBA reactants . In general, the results of both assays were in good agreement . No consistent pattern of interaction between lipid peroxidation, GSH, and enzyme leakage was discernible in the results of the present study . It is suggested that perhaps the best approach to an experimental situation where lipid peroxidation is thought to be of central importance is measurement of both parameters, TBA reactants and ethane concentrations. Arthritis Rheum, 1982 Feb, 25(2), 196 - 203 In vitro conditions affecting the synthesis of sulfated proteoglycans by normal and rheumatoid synovial cells in culture; Marsh JM et al.; In vitro conditions affecting synthesis of sulfated proteoglycans by cell suspensions derived from monolayer cell cultures of normal and rheumatoid synovial tissue were examined . The capacity of cells to synthesize proteoglycans was estimated by the incorporation of 35S--sulfate into cetylpyridinium chloride--precipitable material . Synthesis of sulfated proteoglycans was maximal during log phase, and after 2--3 hours of recovery from disaggregation . Normal synovial cells appeared to be more sensitive to changes in serum concentration than were rheumatoid synovial cells, but rheumatoid synovial cells were more sensitive to changes in cell density . The proportion of newly synthesized extracellular proteoglycans increased with the duration of incubation in 35S--sulfate. Immunology, 1982 Feb, 45(2), 371 - 80 Mast cell growth on fibroblast monolayers: two-cell entities; Ginsburg H et al.; Clonal mast cell differentiation occurs when mesenteric lymph node cells from mice immunized with an antigen are grown in its presence on fibroblast monolayers prepared from mouse embryonic skin . Two types of mast cell clones are identified: the first, originates from a precursor present in the lymphoid cell suspension and the second, from a precursor in the fibroblast monolayer . Clones of the first type fail to appear when T cells are eliminated from the suspension; but they grow luxuriantly in the presence of fluid, harvested from cultures containing the antigen-sensitive T cells exposed to the antigen . The two mast cells differ in the clonal size and rate of growth, life span, cell size, shape and size of the granules and numbers of IgE receptors . It is concluded that the rapidly multiplying lymphoid mast cells associate with the mucosae of the respiratory and gastrointestinal tracts and appear in large number in response to immunological stimuli; the long lived mast cells derived from the embryonic skin monolayer are found in the general connective tissue. Br J Cancer, 1982 Feb, 45(2), 201 - 8 Cells bearing Fc receptors in human malignant solid tumours; Svennevig JL et al.; Fc-receptor-bearing cells forming EA rosettes with antibody-coated human erythrocytes (Ripley) were studied in cell suspensions and in purified preparations of mononuclear cells (MC) from 20 human malignant tumours . The EA rosettes were studied in preparations made by cytocentrifugation and the rosette-forming cells identified by their nonspecific-esterase activity and phagocytic capacity . Fc receptors were found on 16 +/- 20% of all cells in the primary cell suspensions . Significantly more tumour-infiltrating lymphocytes had detectable Fc receptors normal control subjects (14 +/- 6%) . There was a significant correlation between the proportion of lymphocytes lacking T and B markers (null cells) and the proportion of lymphocytes with Fc receptors . Fc receptors were also found on most tumour-infiltrating macrophages, on some T lymphocytes and polymorphonuclear cells and on a smaller percentage of the tumour cells . The significance of the Fc receptor and its usefulness as a marker of "host infiltration" into the tumours is discussed. In Vitro, 1982 Feb, 18(2), 141 - 8 Microcarriers: a new approach to pancreatic islet cell culture; Bone AJ et al.; Free islet cell suspensions were prepared from isolated fetal rat islets using a modified enzyme dispersion technique . The islet cells were dispensed into a culture flask containing microcarriers (Cytodex) suspended in culture medium RPMI 1640 by a slowly rotating bar magnet . Microscopical examination of the beads showed that the islet cells attached and then progressively proliferated on the surface of the beads as a monolayer . A highly sustained release of insulin from the beads to the medium was observed during the 7 d culture period . The functional viability of the cultured islet cells was further demonstrated by the ability of batches of the cell-coated beads to synthesize insulin and to increase the insulin release in response to an acute challenge (16.7 mmol/l glucose plus 5 mmol/l theophylline) . The results suggest that bead microcarriers may provide a new approach to monolayer islet cell culture providing functional monolayers, which can easily be transferred to different test systems and further manipulated. J Clin Pathol, 1982 Feb, 35(2), 191 - 4 Cell structure and percent viability by a slide centrifuge technique; Fitzgerald MG et al.; It was found that a slide centrifuge (Cytospin) preparation of a cell suspension allowed a reliable assessment of not only cell structure but also the percentage of non-viable cells . The non-viable cells appeared as "smear" cells and paralleled in number the cells taking up trypan blue . Direct experiment showed the unstained viable cells in a trypan blue cell suspension remained intact in a Cytospin preparation while the cells taking up trypan blue were the "smear" cells . The non-viability of the "smear" cells was confirmed by their inability to survive in culture. J Clin Pathol, 1982 Feb, 35(2), 139 - 43 Detection of surface immunoglobulins of human lymphoid cells: a comparative study of live and fixed cells using a direct immunoperoxidase procedure; Laurent G et al.; Surface immunoglobulins (Ig) of normal and malignant lymphoid cells were detected on prefixed, smeared (method A) and live (method B) cell suspensions; the results were compared with regard to staining patterns, specificity and sensitivity . In both methods surface Ig were detected by a direct immunoperoxidase procedure using conjugated purified antibody . Although method A has practical advantages, method B is more sensitive . The reasons for this discrepancy are discussed in relation to surface Ig denaturation and redistribution. J Endocrinol, 1982 Feb, 92(2), 293 - 302 Isolation of rat Leydig cells by density gradient centrifugation; Gale JS et al.; A rapid method for preparing Leydig cells from rat testes is described . An interstitial cell suspension, prepared by collagenase treatment of decapsulated testes, was centrifugal for 10 min over a cushion of 60% (v/v) Percoll to remove red blood cells, and then centrifuged for 20 min in a 0-60% linear density gradient of Percoll . Seventy-four per cent of the cells present in that fraction of the gradient comprising 35-50% Percoll were Leydig cells; the yield from each testis was about 1.5 x 10(6) cells . The Leydig cells appeared viable, excluded Trypan blue, possessed high-affinity binding sites for human chorionic gonadotrophin (hCG) and synthesized increased quantities of testosterone in response to hCG . The cells could be stored overnight in 20% (v/v) glycerol at -20 degrees C, with only minimal effect on the specific activities of a number of enzymes used as markers of subcellular components . Testosterone production in vitro by the cells after storage for 20 h was greater than that of hCG-stimulated fresh cells and was not further increased by hCG. Blood, 1982 Feb, 59(2), 226 - 32 In situ immunologic characterization of cellular constituents in lymph nodes and spleens involved by Hodgkin's disease; Poppema S et al.; The cellular constituents in lymph nodes and spleens of patients with Hodgkin's disease were studied with a series of monoclonal antibodies directed against human thymocyte, peripheral T-cell, and la antigens . Utilizing both an immunoperoxidase technique on frozen tissue sections and indirect immunofluorescence on cell suspensions, wer found that a majority of lymphocytes were T cells, since they stained with anti-T1 and anti-T3 antibodies, which react with all peripheral T cells . In addition, most of these cells were reactive with anti-T4 antibody, which defines the helper/inducer T-cell population, whereas only a minority of cells stained with anti-T5 and anti-T8 antibodies, which are reactive with suppressor/cytotoxic T cells . Moreover, a large proportion of T cells expressed T10 antigen, which is found on activated T cells . A minority of the T cells also expressed la antigen(s), again suggesting that some of the T cells are activated . In contrast, the Reed-sternberg cells did not react with any of these anti-T-cell antibodies or with anti-IgM antiserum, but displayed strong membrane and cytoplasmic staining with anti-la antibody . Taken together, these findings suggest that Reed-Sternberg cells are not of T-cell lineage but may be derived from antigen-presenting reticulum cells in the thymus-dependent areas of lymphoid tissues; these cells are normally associated with T4+ cells. J Invest Dermatol, 1982 Feb, 78(2), 125 - 7 Human skin cells synthesize HLA-DR molecules of the same charge and molecular weight as those synthesized by autologous lymphocytes; Morhenn VB et al.; Between 5 and 14% of human skin cells, separated by trypsinization, express HLA-DR antigen as detected by indirect immunofluorescence with monoclonal anti-DR antibody . To determine the source and structure of these DR molecules, skin cell suspensions were biosynthetically labeled with 35S-methionine and the radiolabeled DR molecules were analyzed by the method of two-dimensional polyacrylamide gel electrophoresis . The results indicated the presence of heavy, intermediate, and light chains indistinguishable from those of B lymphocyte DR . Gels in which DR immunoprecipitates from skin cells and autologous lymphocytes were run simultaneously confirmed this finding . Thus, skin cells and lymphocytes synthesize HLA-DR antigen of identical charge and molecular weight. J Natl Cancer Inst, 1982 Feb, 68(2), 325 - 8 Lipopolysaccharide and pristane-induced peritoneal exudate requirements for plasma tumor cell colony formation; Platica M et al.; The role for lipopolysaccharide (LPS) and a pristane-induced peritoneal exudate (PIPE) on the in vitro development of murine plasmacytoma was studied . MOPC-315 cell suspensions showed little tendency for colony formation in a soft agar culture medium . Additions of LPS and PIPE were required for maximal colony formation . The LPS effect was dose-dependent down to nanogram quantities . PIPE prepared from inbred strains of mice not susceptible to pristane-induced plasmacytoma (DBA/2) or from normal peritoneal washings was ineffective . PIPE activity was radioresistant and not transferable by cell-free conditioned medium . Three strains of transplantable plasmacytomas showed colony formation stimulation by LPS plus PIPE, but LPS and PIPE were ineffective with lymphosarcoma P1798. Brain Res, 1982 Feb, 255(2), 167 - 80 GABA uptake and release in purified neuronal and nonneuronal cultures from chick embryo retina; Hyndman AG et al.; Uptake and release of gamma-aminobutyric acid (GABA) have been studied using glia-free, purified neuronal cultures from 8-day chick embryo retina . At 3 days in vitro 65% of the neurons showed high-affinity GABA uptake . These neurons appeared heavily labeled after incubation in 5 X 10(-8) M {3H}GABA, but no labeling was detected when the incubation was carried out at 4 degrees C, or in the absence of Na+ ions . Diaminobutyric acid (DABA) also blocked completely the neuronal uptake of GABA, while beta -alanine was ineffective at similar concentrations . At 6 days in vitro Na+- and temperature-dependent GABA uptake was present in 50% of the neurons . In addition, in 80% of those neurons the uptake was insensitive to DABA or beta -alanine, whereas in the remaining 20% it was blocked by DABA but not by beta -alanine . Important developmental changes were also found in the capacity of the neurons to release GABA into the medium . Spontaneous GABA release (i.e . that taking place in regular medium, containing 5 mM K+) was higher at 3 than at 6 days in vitro . However, increasing the K+ concentration to 56 mM had minimal effects at 3 days in vitro, but induced a 2 to 3-fold increase in GABA release at 6 days in vitro . This K+-induced release appeared to be Ca2+-dependent, since it was substantially reduced the presence of 10 mM Co2+ . Cultures containing a confluent monolayer of nonneuronal flat cells were generated by seeding retinal cell suspensions on poorly adhesive substrata . Retina nonneuronal cells showed, during the first 10 days in vitro, a high-affinity mechanism for GABA uptake which was Na+- and temperature-dependent, and was reduced by 85% by DABA but was practically unaffected by beta-alanine . This uptake mechanism seemed to be lost towards the end of the second week in vitro, and could not be detected after 21 days culture. Proc Natl Acad Sci U S A, 1982 Feb, 79(4), 1166 - 70 ESR studies of O2 uptake by Chinese hamster ovary cells during the cell cycle; Lai CS et al.; Magnetic interactions between dissolved oxygen and nitroxide radical spin probes lead to broadening of the ESR lines . We have used a closed-chamber method based on this property to determine the maximum rate of O2 uptake per cell (Vmax per cell) in cultured mammalian cells . A suitable spin probe and a cell suspension are mixed in an aerated medium, and the rate of disappearance of dissolved O2 is measured . The effects of temperature, pH, and microwave power on the determination of dissolved oxygen in solution were studied . For asynchronous Chinese hamster ovary cells, oxygen uptake is 3.8 X 10(7) oxygen molecules per cell per sec and appears to be enzymatically limited at oxygen concentrations greater than 10 microM . About 5-10 X 10(5) cells were used for each measurement, making it possible to study mitotically synchronized cells . Using this method, we have found that Vmax per unit cell volume changes during the cell cycle of Chinese hamster ovary cells from a minimum in mitosis to maxima in both G1 and late S phases . Advantages and limitations of spin probes for studying the O2 uptake of intact cells are discussed. J Cell Biol, 1982 Feb, 92(2), 505 - 13 Cortical cell populations from rabbit kidney isolated by free-flow electrophoresis: characterization by measurement of hormone-sensitive adenylate cyclase; Vandewalle A et al.; Free-flow electrophoresis allows the separation of different cell populations from a cell suspension isolated from rabbit kidney cortex after perfusion of the kidneys with a calcium-binder, followed by gentle mechanical treatment . After electrophoretic separation, analysis of the adenylate cyclase activities after stimulation by various hormones allows the precise determination of the origin of the cell populations with different electrophoretic mobilities . Adenylate cyclase from the slow-moving main cell population was only sensitive to parathyroid hormone . These cells had also high alkaline phosphatase content, further demonstrating their proximal origin . The various fast-moving cell populations had adenylate cyclase sensitive to isoproterenol and arginine vasopressin but were less sensitive to parathyroid hormone than the slow-moving cells . Their alkaline phosphatase content was also much lower . This indicates that these fast-moving cell populations originate from both the granulous segment of the distal tubule and from the collecting ducts . The adenylate cyclase activity and the cyclic AMP contents of isolated proximal cells maintained in culture medium were also investigated. J Exp Med, 1982 Feb 1, 155(2), 629 - 34 Concomitant induction of the cell surface expression of Ia determinants and accessory cell function by a murine macrophage tumor cell line; Walker EB et al.; This study demonstrates that an uncharacterized soluble factor produced in concanavalin A-induced rat spleen cell suspensions has the capacity to induce the increased expression of cell surface H-2K and H-2D molecules and the expression of I-region gene products on murine monocyte-macrophage lineage tumors that are not Ia positive in the absence of the factor . In parallel with induction of serologically defined Ia specificities, Ia-induced WEHI-3 macrophage tumor cells are capable of providing accessory cell function in stimulating IL-2 production by T-T hybridomas that are activated in a major histocompatibility complex-restricted, antigen-dependent fashion . The uninduced Ia-negative WEHI-3 tumor cells do not trigger a comparable response in this assay system. Exp Hematol, 1982 Feb, 10(2), 217 - 27 Marrow microenvironment transfer by heterotopic transplantation of freshly isolated and cultured cells in porous sponges; Friedenstein AJ et al.; Cells for study were obtained from mouse bone marrow by three methods: 1) Mechanical dissociation; 2) Trypsin release; and 3) Passage through a syringe and needle . The ratio of the fibroblast colony-forming cells in these suspensions was, respectively, 1:13:1 . Of the colonies formed by cells obtained by trypsin release, 70% were alkaline phosphatase positive fibroblasts . Freshly isolated marrow cells and cells harvested, by trypsin treatment, from cultures established from the differently obtained bone marrow cell suspensions, were absorbed into porous sponges for transplantation under the renal capsule of mice . Ossicles containing bone marrow formed after transplantation of either freshly isolated cells, or cultured cells obtained from marrow by trypsin treatment, but not after the transplantation of cells cultured from marrow obtained by mechanical dissociation . The size of the bone marrow "organs" that formed was determined by both the number of cells grafted and the size of the sponge in which they had been absorbed for grafting. Biochim Biophys Acta, 1982 Jan 22, 684(2), 228 - 32 Phloretinyl-3'-benzylazide: a high affinity probe for the sugar transporter in human erythrocytes . II . Irreversible transport inhibition is induced by photolysis; Fannin FF et al.; In the dark, phloretinyl-3'-benzylazide (PBAz), at a nominal concentration of 10 microM, will inhibit the transport of D-glucose in human erythrocytes by more than 90% . This inhibition can be completely reversed by percolating the cell suspension through a small column of Sephadex G-10; cells recovered after this treatment, and then loaded with 100 mM D-glucose, possess a transport capacity (glucose efflux) equal to untreated cells . The Sephadex matrix completely removes non-covalently bound inhibitor even though, under these conditions (subdued light, 0.2% hematocrit, 0 degrees C, pH 6.2 or 7.8), from 70 to 80% of the PBAz added is bound to the cells (mostly non-specifically to hemoglobin) . However, when erythrocytes exposed to 10 microM inhibitor are irradiated with long wavelength ultraviolet light, the glucose transporter is irreversibly inhibited; after 1 min irradiation, about 50% of transporter activity cannot be restored by Sephadex treatment . Under identical conditions, control cells (no PBAz, but irradiated and treated with Sephadex) retain over 90% of carrier activity . The photolytic conversion of the inhibition to an irreversible form is directly dependent on PBAz concentration . The results reaffirm our earlier conclusions that PBAz is a potentially useful photoaffinity labeling agent for the glucose transporter in erythrocyte membranes. Fortschr Med, 1982 Jan 21, 100(3), 83 - 7 {Transplantation of isolated chondrocytes in articular cartilage defects . Regeneration of adult hyaline cartilage with fetal chondrocytes}; Helbing G; Viability and proliferative capacity of the chondrocytes are particularly important for a successful cartilage graft . This can be demonstrated by a new in vitro colony-forming assay . Only fetal or neonatal chondrocytes are considered to be applicable according to these criteria . Single cell suspensions prepared from articular cartilage can be transplanted without technical problems . In animal experiments articular cartilage regenerated after transplantation of neonatal chondrocytes, whereas only connective tissue was found in controls. Biochim Biophys Acta, 1982 Jan 12, 714(1), 157 - 63 Cholinergic muscarinic stimulation of steroidogenesis in bovine adrenal cortex fasciculata cell suspensions; Hadjian AJ et al.; Acetylcholine was found to acutely stimulate cortisol production by bovine fasciculata adrenocortical cell suspensions . This effect was maximal at 10(-4) M acetylcholine concentration, resulted in a 5-fold increase in cortisol production over the control after 1 h incubation, and represented about one fifth of the ACTH maximal stimulation under the same conditions . Acetylcholine-stimulated steroidogenesis was concentration-dependent (10(-8)-10(-5) M), proportional to the cell number (5 X 10(5)-1 X 10(6)) and reached a plateau after 30 min incubation . Use of various cholinergic specific agonists and antagonists showed that the steroidogenic action of acetylcholine was a typical muscarinic effect . This character is in agreement with the previously demonstrated presence of muscarinic receptors in bovine adrenocortical tissue . The steroidogenic effect of acetylcholine required the presence of extracellular calcium in the medium and was impaired upon addition of tetracaine and procaine . No change in cyclic AMP nor cyclic GMP levels could be detected in the system under acetylcholine stimulation . Acetylcholine appeared to exhibit a synergistic effect in combination with ACTH, and exogenous cyclic AMP; these observations suggest a different mechanism of action for acetylcholine and ACTH and point to a possible cholinergic participation in the regulation of adrenocortical differentiated functions in vivo. Life Sci, 1982 Jan 4, 30(1), 85 - 91 Regulation of tyrosine hydroxylase activity in retinal cell suspensions: effects of potassium and 8-bromo cyclic AMP; Iuvone PM et al.; Tyrosine hydroxylase (TH) contained in dopamine (DA) neurons of rat retina is activated in vivo as a consequence of photic stimulation . Experiments were conducted to test the effects of changes of membrane potential and of cyclic AMP-dependent protein phosphorylation on TH activity of these retinal neurons . Retinas were dissociated into suspensions of apparently viable cells to test the direct effects of pharmacological manipulations on TH activity in the absence of trans-synaptic influences . To test the effects of changes of membrane potential on TH activity we examined the effects of a depolarizing agent, potassium . Incubation of cell suspensions in Ringer's solution containing a depolarizing concentration of K+ (52 mM) resulted in a significant increase of TH activity, suggesting that membrane depolarization may trigger a series of molecular events that leads to TH activation . Incubation of cell suspensions in the presence of 8-bromo cyclic AMP, a cyclic AMP analog that is known to activate cyclic AMP-dependent processes following extra-cellular application, resulted in a significant activation of TH that was comparable to that produced in vitro by cyclic AMP-dependent protein phosphorylation . These data support the hypothesis that membrane potential plays a role in the regulation of TH activity, and indicate that cyclic AMP-dependent phosphorylation can activate retinal TH in situ . The apparent viability of the retinal cells in suspension suggests that this preparation may be useful for studying synaptic regulatory mechanisms. Prostate, 1982, 3(3), 253 - 75 Intraprostatic and subcutaneous transplantation of a spontaneous prostatic carcinoma (11095) to male Fischer rats (F344): an ultrastructural study; Weber P et al.; A prostate carcinoma arising spontaneously in a Fischer rat was transplanted by cell suspension to male and female Fischer rats . Tumour growth was rapid in both subcutaneous and intraprostatic sites of inoculation in males and subcutaneously in females . The growth rate was greater in male than in female hosts and the tumour developed as a solid form in both groups . It had a heterogeneous cell population with ultrastructural characteristics similar to basal cells of the normal prostate and to tumour cells seen in human prostate adenocarcinoma . Later stages of tumour growth transformed it to a cornifying squamous cell carcinoma . The morphological description of tumours produced by transplantation provides the basis for further studies using this method as a model for investigating the response of prostate carcinoma to endocrine manipulation. Physiol Chem Phys, 1982, 14(4), 381 - 4 1H-NMR spectroscopic study of aerobic glucose metabolism in Toxoplasma gondii harvested from the peritoneal exudate of experimentally infected mice; Ohsaka A et al.; 1H-nuclear magnetic resonance (NMR) was used for the study of aerobic glucose metabolism in Toxoplasma gondii harvested from the peritoneal exudate of experimentally infected mice . Without any pretreatment (i.e., extraction, separation or purification) of the supernatant fluids from the cell suspension incubated with glucose for various time intervals, we were able to identify and quantitate the end-products of glucose metabolism by means of 1H-NMR . The major end-products found were lactic acid and acetic acid. Cancer Metastasis Rev, 1982, 1(1), 65 - 81 NK cell-target interactions: approaches towards definition of recognition structures; Urdal DL et al.; NK cells lyse an uncommonly wide range of cell types, implying either that they (the NK cells) have clonally distributed receptors each of which is capable of interacting with a very limited number of cell types or, alternatively, that susceptible target cells share a common characteristic . A number of experimental approaches have suggested that the cytotoxic 'specificity' of NK cells is not clonally distributed . Thus, clones of NK cells, established from mouse spleen cell suspensions, showed no greater restriction in the spectrum of target cells which they could lyse, than did the parent spleen cell populations from which they were derived . It seems likely therefore that the wide range of target cell types that can be lysed by NK cells share common cell surface characteristics which render them susceptible . As lysis results from membrane-membrane interactions, it seemed logical that a search for 'hallmarks' of NK susceptibility should begin with a detailed examination of the plasma membrane of susceptible cells . Analysis of one pair of lymphoma cell variants, selected on account of their markedly different susceptibility to NK cells, suggests that cell surface glycoconjugates may be of significance in determining those effector cell-target cell interactions that lead to lysis . This review outlines attempts to characterize such glycoconjugates. Bioelectromagnetics, 1982, 3(4), 453 - 66 Effects of X-band microwave exposure on rabbit erythrocytes; Cleary SF et al.; Rabbit erythrocytes were exposed in vitro to continuous wave (CW) and pulse-modulated X-band microwaves in wave guide exposure chambers . Erythrocytes were exposed as whole (heparinized) blood suspensions or as washed cells in 1:1 isotonic buffered K+-free saline suspensions . Statistically significant increases in K+ efflux relative to thermal controls were detected when red cells were exposed in whole blood suspensions to either CW or pulsed 8.42-GHz microwaves at SARs that resulted in equilibrium sample temperatures of approximately 24 degrees C . Under the same exposure conditions, no statistically significant K+ efflux occurred in the case of 1:1 red cell suspensions . Measured differences in sample heating rates and temperature gradients between microwave-exposed and heated control suspensions may account in part for the differential effect of microwave exposure but such effects do not appear to explain the results of this study fully. Arkh Patol, 1982, 44(10), 3 - 11 {Stromal bone marrow cells and the hematopoietic microenvironment}; Fridenshtein AIa; Stromal clonogenic fibroblasts of the bone marrow possessing osteogenic potential are responsible for hemopoietic bone marrow microenvironment . These cells may be recovered from the bone marrow by cloning in cell cultures; their progeny cells upon retransplantation into the body form new bone marrow organs . Microenvironmental cells are histogenetically independent of hemopoietic cells . In bone marrow cell suspensions prepared by bone marrow tissues trypsinization stromal clonogenic cells are present in greater concentrations than in those prepared by mechanical mincing of the bone marrow tissue . New ectopic bone marrow organs may be created by transplantation of trypsinized suspensions in porous sponges. Res Exp Med (Berl), 1982, 181(2), 147 - 54 An in vitro model for the study of human parathyroid gland tissue: single cell suspensions and monolayer cultures; Wagner PK et al.; An in vitro model for studies of parathyroid physiology is described using single cell suspensions and adherent monolayer cultures of human parathyroid tissue . The isolated cells were viable and maintained functional properties tested by calcium and magnesium sensitivity . Parathyroid hormone (PTH) secretion could be suppressed by rising calcium and magnesium concentrations . The secretory behaviour of the cells was not altered by cultivation or cryopreservation . The morphological integrity of the individual cells after mechanical and enzymatic preparation was confirmed by light and electron microscopy. Physiol Chem Phys, 1982, 14(1), 13 - 7 Kinetics of the reaction of carbon monoxide and oxygen with T-state hemoglobin in human red blood cell suspensions studied by dye laser flash photolysis; Hasinoff BB; The reaction kinetics of carbon monoxide and oxygen binding to slow reacting T-state "deoxy" hemoglobin (HbT) in human red blood cell (RBC) suspensions were studied with a 300-ns pulse width dye laser to photodissociate either HbCO or HbO2 . The rate constants were determined using only the first 10% of the slow HbT reaction, as at longer times diffusion of ligand away from the RBC seriously affects the kinetics . The rate constants measured are similar to those in solution, indicating that the different physical environment in the RBC has little effect on the Hb kinetics. Cell Tissue Res, 1982, 227(1), 225 - 9 Presence of adrenal medullary chromaffin cells in primary adrenal dispersions and their persistence in long-term culture; Moore NA et al.; Cell suspensions prepared by enzymatic dispersion of whole rat adrenal glands for the purpose of studying adrenocortical cells were found to contain chromaffin cells even though they are commonly thought to not survive in such preparations . These cells fluoresced when treated with methods specific for catecholamines . The fluorescent cells persisted in the cultures of "cortical" cells, took on the morphology of neurons in the cultures, maintained their specific catecholamine fluorescence in long-term cultures, and ultrastructurally were identical to chromaffin cells. Acta Biol Med Ger, 1982, 41(11), 1061 - 73 Quasistationary states in red cell suspensions with special regard to blood and red cell preservation; Wolf J; A biophysical model of a red cell suspension which is considered as a closed two-compartment system is established . The model includes the main components of resuspending solutions for red cell preservation which are of biophysical importance . Knowing the composition of the resuspending solution and the mixture cell sediment/resuspending solution the relative cell volume, the cell pH, the pH of the medium, and the transmembrane potential can be calculated . The model is in good accordance with experimental findings (temperature dependence of suspension pH, depolarisation of cells by addition of sucrose or citrate to the outer medium, acidification of the medium by lactate production) and enables one to explain these observations to a certain degree of accuracy also quantitatively . Relations to biochemical aspects of red cell and blood preservation are shown. Acta Chir Scand, 1982, 148(7), 609 - 12 Interruption of hepatic arterial supply in rats with liver tumors; Carlsson G et al.; Liver tumors receive their main blood supply from the hepatic artery . Various types of interruption of arterial blood supply to the liver are considered to give a reduction in liver tumor growth . Inoculation of a tumor cell suspension of a transplanted benzpyrene-induced sarcoma was used to induce a tumor in the liver of Lister-hooded rats . The influence of hepatic artery ligation (HAL) and regular liver dearterialization, i.e . dividing of all structures to the liver except portal vein, common bile duct and hepatic veins, was tested against control and sham operation . Ligation of the hepatic artery gave a statistically significant temporary reduction of liver tumor growth . A continuous growth of the tumors was found after regular liver dearterialization, sham procedure and in the controls . Seven and 14 days after the procedures this difference between HAL and regular liver dearterialization was recognized statistically significant (p less than 0.001) in groups with only five animals. Res Exp Med (Berl), 1982, 181(3), 205 - 10 {Effect of calcium and magnesium on parathyroid hormone release from human parathyroid tissue in vitro}; Wagner PK et al.; The effects of calcium and magnesium on parathyroid hormone release from eight adenomas causing primary hyperparathyroidism and six hyperplastic glands causing hypercalcemic secondary hyperparathyroidism were investigated in vitro using single cell suspensions from the respective tissue . We observed suppression of parathyroid hormone release with increasing concentrations of either cation . The quantitative hormone secretions of both adenomatous and hyperplastic glands was identical. Blood Cells, 1982, 8(2), 315 - 28 Rheological assessment of antisickling effects of pyridoxine and pyridoxal; Kuranstin-Mills J et al.; The antisickling effects of pyridoxine and pyridoxal on intact sickle erythrocytes (SRBCs) were assessed by microviscometry of cell suspensions at shear rates of 1.15 to 230.0/s, measurement of cellular deformability by cell filtration and scanning and transmission electron microscopy (EM) . Incubation of fresh SRBCs in albuminated (0.1%) phosphate buffered saline, pH 7.4, with 5 to 30 mM, pyridoxine or pyridoxal at 37 degrees C for 90 min followed by deoxygenation to pO2 = 25 mmHg, decreased the percentage of sickled cells observed as a function of vitamin concentration . EM of fresh SRBCs incubated with 20 mM pyridoxine or pyridoxal at 25 mmHg pO2 showed mostly discocytes without intracellular fibers indicating absence of hemoglobin polymerization . Determination of fluidity (viscosity-1) versus shear stress showed that both pyridoxine and pyridoxal significantly (P less than 0.01) increased the fluidity of a deoxygenated suspension of SRBCs . The estimated apparent yield stress from Casson plots for the vitamin-treated deoxygenated cells and the deoxygenated control cells were 0.11 and 0.18 dynes/cm2 respectively . The relative resistance of 0.2% cell suspension to flow through 5 microns Nuclepore filters indicated a significant increase (greater than 75%) in the deformability of the vitamin-treated deoxygenated sickle cells . The ultrastructure of the SRBCs through the filter pores further suggested that the fluidity of the intracellular milieu of the vitamin-treated deoxygenated cells was higher than that of the deoxygenated controls . These data support the hypothesis that pyridoxylation of sickle hemoglobin inhibits sickling, increases the fluidity of sickle hemoglobin under low pO2, and thereby enhances the deformability of SRBCs in models of capillary blood flow. Blood Cells, 1982, 8(2), 263 - 72 Sickling as a function of oxygen delivery: effect of simulated transfusions of stored, fresh and inositol-hexaphosphate-loaded (low affinity) red cells; Kumpati J et al.; Low oxygen affinity red cells were prepared by incorporating inositol hexaphosphate (IHP) into red blood cells (rbc) by means of a liposomal transport system . The effect of in vitro simulated exchange transfusions on sickling was studied with buffered red cell suspensions containing 50% SS cells and 50% test cells . Test rbc were either stored cells with high oxygen affinity, fresh cells with normal affinity or IHP-loaded cells with decreased affinity . Oxygen equilibrium curves and percentage sickling as a function of PO2 were determined and the data analyzed in terms of percentage sickling as a function of oxygen delivery . Our simplified analysis shows that simulated exchange transfusion with stored and, to a lesser extent even with fresh blood, results in a decreased venous PO2 and increased sickling of the remaining SS cells . In contrast, transfusion with IHP-loaded cells results in higher venous PO2 values and less sickling throughout the range of oxygen delivery . Thus, the transfusion of IHP-loaded cells may result in less sickling of the remaining SS cells in addition to the normal dilutional effect. Exp Pathol, 1982, 22(3), 179 - 80 Induction of a slowly growing transplantable adenocarcinoma metastasizing into the lungs; Dutter A et al.; In male Long Evans rats Dibutylnitrosamin induces adenocarcinomas of the lung . Transplantation of a tumor cell suspension prepared from the tumor leads in 100% of cases to death by metastases from adenocarcinoma of the lung after an average of 24 weeks when applied i.m . in a syngeneic system. Comp Biochem Physiol B, 1982, 73(4), 829 - 33 The mitochondrial function in hemosome formation and hemoglobin biosynthesis; Brunner A Jr et al.; 1 . Rabbit-kidney epithelial cell cultures were induced to synthesize hemoglobin, by previously mixing cell suspensions with solutions containing reticulocyte free globin, hemoglobin and anemic rabbit blood plasma . As control, a solution without globin was used . 2 . After a 24 hr culture growth period, hemoglobin was absent, as stated through electrophoresis, suggesting hemoglobin denaturation: mitochondria interacted with the incorporated material and particles resembling ferritin molecules were found within 48 hr . 3 . Mitochondria modified remarkably giving rise to lamellated bodies which recomposed to form prohemosomes, presumably containing globin and newly synthesized heme: hemoglobin was still absent up to 72 hr . 4 . After 96 hr hemosomes developed and hemoglobin, apparently constituted by reticulocyte globin, was detected. Stem Cells, 1982, 1(4-5), 261 - 8 2'5'-adenylate inhibition of erythropoietin-dependent colony formation; Orlic D et al.; The tetramer core of 2'5'-adenylate was tested in vitro for its effect on erythroid colony-forming units (CFU-E) from bone marrow of adult rats . Inhibition of CFU-E was 2'5'-adenylate dose dependent, when tested with concentrations of 0.05- 1.0 mM . Also, 2'5'-adenylate was inhibitory when incubated with 2, 6 or 10 X 10(4) cells/0.1 ml plasma clot . At 0.1 mM concentration of 2'5'-adenylate there was a 30-50% reduction of erythroid colonies for each of these three cell concentrations . There was an early decrease in 3H-thymidine incorporation when 2'5'-adenylate was added to erythropoietin-stimulated bone marrow cell suspensions prior to plating . It is suggested that in situ production of 2'5'-adenylate may represent a regulatory mechanisms in red cell maturation. Int J Tissue React, 1982, 4(2), 91 - 4 A comparison of three methods of in vitro culture of human oesophageal mucosa; Simon BG et al.; Three methods of in vitro culture of human oesophageal epithelium were assessed which included the Bijou bottle, cell suspension and organ culture . The epithelium did not survive in the Bijou bottle and the cell suspension caused a growth of oesophageal fibroblasts . Organ culture proved to be the best method with survival up to ten days . Organ culture of human oesophageal epithelium provides an easy in vitro method of studying various cytotoxic factors which may play a role in reflux oesophagitis. Blood Cells, 1982, 8(1), 89 - 101 Nonspecific rheological abnormalities in sickle cell disease; Schmid-Schonbein H et al.; The rheology of blood from normal healthy subjects, from sickle cell patients and hybrid red cell suspensions prepared by mixing HbSS cells with normal plasma and HbA cells with plasma of sickle cell patients were studied . The microrheological behavior was observed in a rheoscope, in which the kinetics of red cell aggregation were also studied systematically . The viscosities of plasma, sera and whole blood samples were determined in a coaxial cylinder and a cone-plate viscometer . The data show that in sickle cell disease a number of nonspecific factors capable of interfering with blood fluidity are changed . These include an elevation of the tendency to red cell aggregation (which also persists after the removal of plasma fibrinogen), the enhanced tendency to red cell aggregation (measured photometrically and by extrapolation of the viscometric data plotted according to the Casson equation) and consequently strong decrease in apparent fluidity, especially at low shear rates . These unspecific abnormalities persist even after deoxygenation and complicate the specific effect of HbSS cells following hemoglobin gelation . Comprehensive pathophysiological hypothesis about circulatory disturbances in sickle cell patients is given which includes concepts about the interaction of general and local hemodynamics and of specific and nonspecific hemorheological determinants of apparent blood fluidity. Blood Cells, 1982, 8(1), 53 - 64 Viscoelastic properties of sickle cells and hemoglobin; Chien S et al.; The quantitative relationship between deoxygenation and rheological behavior of SS cell suspensions and concentrated HbS solutions has been studied under steady shear (viscosity eta) and oscillatory shear (complex viscosity with viscous component eta' and elastic component eta") . decrease of O2 saturation below 80-85% causes eta, eta' and eta" to increase progressively in SS cell suspensions and HbS solutions . These rheological parameters do not change in AA cell suspensions and HbA solutions following deoxygenation . The deoxygenation-induced increase in eta" of HbS solutions from the unmeasurable level when oxygenated reflects the gelation of HbS . At high O2 saturations, the rheological behavior of the intracellular fluid has relatively insignificant contribution to that of the SS cell suspension . As O2 saturation decreases, the viscoelastic properties of the SS cell suspension become increasingly dominated by that of the intracellular HbS . at a given degree of deoxygenation, eta, eta' and eta" of HbS solutions decrease with increasing shear rate, indicating that HbS aggregates can be dispersed by shear stress. Ultrasound Med Biol, 1982, 8(3), 299 - 309 The exposure vessel as a factor in ultrasonically-induced mammalian cell lysis--II . An explanation of the need to rotate exposure tubes; Church CC et al.; This paper is an attempt to explain the need to rotate a polystyrene tube containing a cell suspension in order to obtain cell lysis . Calculations, based on known physical laws, were performed in order to determine the important forces on cells and bubbles and the movements and interactions which these forces are likely to cause . These calculations support the following conclusions: (1) in the absence of rotation, cells and bubbles larger than resonance size are trapped at pressure minima while bubbles smaller than resonance size are trapped at pressure maxima, (2) at 1 W/cm2 with rotation, lysis is caused by cells sweeping through arrays of trapped small bubbles, (3) at higher intensities lysis is caused by both trapped and non-trapped small bubbles. Biorheology, 1982, 19(1/2), 237 - 44 Modifications of the erythrocyte deformability alter the effect of temperature on the relative viscosity of human blood; Rogausch H; The relative viscosity of normal red cell suspensions is independent of temperature at high shear rates . The relative viscosity of suspensions with normovolemic sphered erythrocytes having a reduced deformability, however, is higher at 37 degrees C then at 20 degrees C . It is concluded that the changes of the lipoprotein configuration within the erythrocyte membrane which are proposed to be involved in the disc-sphere-transformation of the red cell, depend on temperature and are responsible for the increased relative viscosity at 37 degrees C. Vopr Onkol, 1982, 28(4), 42 - 4 {Effect of pulsed laser radiation on the lysosomes of Ehrlich ascitic tumor cells}; Moskalik KG et al.; The results of a study on the effect of neodymium pulsed laser radiation on Ehrlich ascites tumor lysosomes are discussed . Tumor cell suspension was exposed in vitro to 10 pulses of laser irradiation with fluences of 10, 25 and 100 J/cm2, at a wavelength of 1.06 microns and a pulse duration of 1 microsecond . A rise in the free and total activities of acid phosphatase in the fraction of isolated lysosomes was registered one or four hours after exposure, particularly, at fluences of 10 and 25 J/cm2 . However, laser radiation did not affect acid phosphatase activity in preparations of solubilized enzyme . Special experiments involving the use of detergents showed the increased activity of acid phosphatase in isolated lysosome fraction to be due to labilization of cytoplasmic membranes of tumor cells. Acta Pharmacol Toxicol (Copenh), 1982 Jan, 50(1), 67 - 74 Interaction between drug acetylation and ethanol, acetate, pyruvate, citrate, and L(-) carnitine in isolated rat liver parenchymal cells; Olsen H; The acetylation of sulfanilamide and procainamide in suspensions of isolated rat parenchymal cells was studied in absence and presence of ethanol (33mM), citrate (4 mM), pyruvate (4 mM), and L(-)carnitine (2 mM) . Ethanol treatment enhanced the sulfanilamide acetylation whereas the acetylation of procainamide was considered to be unchanged . Acetate (1-5 mM), citrate, and pyruvate treatment enhanced the acetylation of both sulfanilamide and procainamide . Acetate (4 mM) increased both Km and Vmax of both sulfanilamide and procainamide acetylation . Combined treatment with L(-)carnitine and either acetate, pyruvate, or citrate enhanced the acetylation rate of sulfanilamide more than acetate, pyruvate, or citrate, respectively alone . In cell suspensions treated with L(-) carnitine and acetate or pyruvate, the acetylation kinetics of sulfanilamide changed from zero-to apparent first-order . With procainamide as test drug, a further increase of the acetylation rate was found when L(-) carnitine was added to citrate pyruvate . Acetyl-CoA increased the rate of sulfanilamide acetylation in rat liver homogenates in a dose dependent manner. J Cancer Res Clin Oncol, 1982, 102(3), 253 - 63 Radiation effect on partially synchronized Yoshida sarcoma cells; Bippus PH et al.; Yoshida sarcoma cells, which have the same growth characteristics as ascites cells in the mouse and in cell suspension, were partially synchronized in vitro by means of excess thymidine (0.1 mM thymidine for 18 h) . The growth of non-synchronized cultures was inhibited by irradiation, the degree depending on the dose of radiation . At the same time, a 50% inhibition in vivo (380 rad) and in vitro (480 rad) was determined . The incorporation of 3H-thymidine into the DNA is inhibited by 10-32%, depending on the radiation dose . The mitotic index decreases 2 h after irradiation by a dose-dependent amount . A mitotic maximum develops later; the delay is dose-dependent . Partially synchronized cells were irradiated in the G1/S-, G2-, and G1-phase . As compared to the 3H-thymidine incorporation and the mitotic index there were no significant differences between the cultures which were irradiated in the individual phases of the non-synchronized control cultures . The cultures which were irradiated in the G2-phase, however, showed a significantly reduced growth in vivo after 48 h . If the cells were cultured for more than 72 h after irradiation, the differences between the cultures irradiated in the G2-phase and the other phases were reduced. J Membr Biol, 1982, 65(1-2), 139 - 45 Effects of salicylate on HCO-3/Cl- exchange across the human erythrocyte membrane; Crandall ED et al.; Changes in extracellular pH (pHo) in human red cell suspensions were monitored in a stopped-flow rapid reaction apparatus . A 20% suspension of washed human RBC in saline at pH 7 containing NaHCO3 and extracellular carbonic anhydrase was mixed with an equal volume of buffered saline solution at pH 6.7 . Sodium salicylate, when present, was added to both the erythrocyte suspension and the buffer solution . The effects of salicylate in the therapeutic to toxic concentration range on HCO-3/Cl- exchange were studied at 37 degree C . HCO-3/Cl- exchange flux was estimated using the extracellular buffer capacity and the difference between dpHo/dt using a control RBC suspension and that using a suspension of RBC whose anion exchange pathway was markedly inhibited . The results show that salicylate competitively decreases the rate of HCO-3/Cl- exchange, with inhibition increasing as salicylate concentration increases . KI is approximately 2.4 mM . At a salicylate concentration of 10 mM, HCO-3/Cl- exchange under the conditions of our experiments was inhibited by more than 70% . These findings are consistent with the possibility that CO2 transfer in capillary beds in vivo may be diminished in the presence of salicylate due to slowing of red cell HCO-3/Cl- exchange. J Pathol, 1982 Jan, 136(1), 1 - 13 Isolated epithelioid cells from disaggregated BCG granulomas - an ultrastructural study; Williams GT; Non-caseating epithelioid granulomas have been induced in rats by the subcutaneous injection of BCG vaccine and their cellular disaggregation to yield isolated epithelioid cells has been achieved using collagenase . Ultrastructurally epithelioid cells in both intact granulomas and disaggregated cell suspensions showed a spectrum of appearances ranging from cells with conspicuous rough surfaced endoplasmic reticulum ("plasmacytoid epithelioid cells") to cells containing numerous cytoplasmic membrane-bound vesicles ("vesicular epithelioid cells") . Numerous intermediate forms were present . Endocytosed material was inconspicuous . Investigation of disaggregated 4, 10 and 28 day lesions showed that the proportion of vesicular epithelioid cells increased with maturation of the granulomas . Nevertheless a full spectrum of epithelioid cell morphology was present as early as 4 days . It is suggested that mononuclear phagocyte cells entering the granuloma transform into vesicular epithelioid cells via an intermediate plasmacytoid stage . The successful isolation of viable epithelioid cells from granulomas may allow the function of these cells to be evaluated further. Invest Ophthalmol Vis Sci, 1982 Jan, 22(1), 115 - 8 Metastasis in a rabbit choroidal melanoma model; Liu LH et al.; A substrain of Greene hamster melanoma adapted to grow in the choroid of the rabbit eye is shown to have the tendency to metastasize . Twenty-one normal rabbits were studied for the effects of vascular inoculation of a tumor cell suspension prepared from this choroidal melanoma, and two rabbits were used for long-term observation of the natural untreated course of the choroidal melanoma . In both groups, metastases to the liver, lung, kidney, and other organs were demonstrated. Int Arch Allergy Appl Immunol, 1982, 67(4), 335 - 9 Contact-dependent expression of actin in chicken lymphocytes in vitro; Woods GM et al.; The presence of actin in chicken bursa, thymus and spleen cells has been confirmed using quantitative neutralization absorptions of anti-actin antibody (AAA) with frozen-thawed preparations of these three cell types . Spleen cells were more effective than thymus or bursa cells in reducing the AAA titre . Cell suspensions of these organs were allowed to settle onto a glass slide, air-dried and stained with AAA . Immunofluorescence was restricted to cell borders at points of contact with other cells . This staining was reversibly inhibited by cytochalasin B but not colchicine . The higher content of actin in spleen cells demonstrated by the absorption experiments was reflected in their more rapid expression of actin during smear formation, and the relative resistance of this to cytochalasin B treatment, compared to thymus and bursa preparations . These findings further support the proposition that actin expression is a function of lymphocyte maturation. Tsitologiia, 1982 Jan, 24(1), 41 - 6 {Albumin synthesis by mouse liver cells}; Borovkov AIu et al.; The presence of albumin in mouse liver cells was examined using the immunoperoxidase method with the help of monospecific antibodies to mouse serum albumin . All the experiments were carried out on the cell suspension . The rat cells were used as a control in the mixture of rat and mouse liver cells . All mouse hepatocytes were shown to contain serum albumin . Our results are compared with those reported by other authors . The advantages of our method over the routine histological technique are discussed. Exp Hematol, 1982 Jan, 10(1), 26 - 35 The organization of hemopoietic tissue as inferred from the effects of 5-fluorouracil; Hodgson GS et al.; Mouse bone marrow obtained one day after injection of 5-fluorouracil (FU) had a markedly diminished content of spleen colony forming units (CFUs) but retained its capacity to repopulate the marrow granulocyte-macrophage colony forming cell (GM-CFC) and CFUs compartments of 850 R irradiated hosts and had only a slightly reduced platelet repopulating ability (PRA) . A significant correlation (r = 0.94, P less than 0.001) was observed between the content of high proliferative potential granulocyte-macrophage progenitor (HPP-GM-CFC) and the platelet and marrow GM-CFC repopulating abilities of bone marrow cell suspensions . Spleens of irradiated mice, injected with marrow from donors treated with FU between 1 and 7 days before showed an increase in colony numbers with time of sampling between 8-13 days after transplantation . In contrast, the colony counts observed in mice injected with normal bone marrow remained constant over that time interval . The colonies derived from bone marrow of FU treated mice grew faster than those from bone marrow of normal mice . Spleens obtained from irradiated mice, 10 days after injection of bone marrow derived from donors treated with FU 1 or 3 days before, showed only a few macroscopic surface colonies but when sectioned were found to contain large numbers of microscopic colonies, 80% of which were megakaryocytic . The results are interpreted on the basis of a clonal succession model of hemopoiesis with stem cells of varying proliferative potential and proliferation rates increasing as capacity for cell production decreases. Adv Exp Med Biol, 1982, 149, 91 - 6 Functional antigen binding by the defective B cells of CBA/N mice; Snippe H et al.; CBA/N mice have an X-linked B cell defect which prevents them from responding to nonmitogenic thymic independent (TI-2) antigens such as dinitrophenylated DNP-Ficoll (1,2) . The F1 male progeny of CBA/N female mice express the same defect . Spleen cell suspensions from such defective mice (CBA/N X C3H/HeN F1 males) could not respond to DNP-Ficoll following in vitro immunization and subsequent transfer into irradiated, syngeneic, F1 male recipients as expected . In contrast, normal CBA/N X C3H/HeN F1 female spleen cells could respond and effect a "rescue"; they mounted strong plaque-forming cell responses 7 days after in vitro exposure to DNP-Ficoll and subsequent transfer into irradiated F1 male recipients . Defective F1 male spleen cells, however, could bind significant quantities of 125I-DNP-Ficoll after in vitro exposure . Extensive washing of these spleen cells could not reverse this binding . Such DNP-Ficoll-exposed and washed F1 male spleen cells could, after transfer, aid normal untreated F1 female cells in their rescue function . The defective F1 male spleen cells could convey immunogenic quantities of DNP-Ficoll to the "rescuing" F1 female cells . Mitomycin treatment of F1 male cells did not interfere with their conveyor function . Goat anti-mouse mu serum impeded the passive antigen conveyor function of defective F1 male cells as did prior exposure to high concentrations of free DNP hapten . Our data support the view that the B cell defect of CBA/N X C3H/HeN F1 male mice does not relate to antigen binding, but rather to an inability to be effectively triggered by certain cell-bound polymeric antigens. Stem Cells, 1982, 1(4-5), 209 - 23 Interaction between macrophages and human tumor clonogenic cells; Hamburger AW et al.; We have demonstrated that autologous macrophages enhance the growth of human tumor cells in soft agar . In an attempt to identify the macrophage subpopulation responsible for enhancing tumor growth, we depleted adherent cells of macrophages bearing surface Ia antigens, and assessed the ability of the residual cell population to support tumor growth . Adherent cells constituted 18-36% of cells from 20 effusions tested . A variable proportion of these adherent cells (33-60%) were lysed by anti-Ia-like antibody and complement . These was no correlation between cloning efficiencies of cells in the original cell suspension and the percentage of Ia-positive adherent cells initially present . Clonogenicity was reduced in all cases by removal of adherent cells . Adherent cells, depleted of Ia-positive macrophages, were able to enhance the growth of nonadherent tumor cells . The growth enhancement was dependent on the number of Ia-negative cells added . The number of colonies grown in the presence of Ia-negative macrophages was between 34 and 350% greater than the number of colonies observed when untreated adherent cells were added in 17 of 21 cases . The effect of the Ia antibody on the functional activity of the adherent cells was complement and dose dependent . Both heteroantibodies and monoclonal antibodies produced a similar effect . The results indicate Ia-negative macrophages could enhance the growth of human tumor cells in soft agar . In addition, Ia-positive macrophages may have suppressed the growth of clonogenic tumor cells Ia-positive macrophages may have suppressed the growth of clonogenic tumor cells. Int Arch Allergy Appl Immunol, 1982, 68(4), 392 - 6 Differential capacity of macrophages from various sources to act as hapten-specific stimulator cells in vitro; von Blomberg M et al.; Lymphocytes from 2,4-dinitrochlorobenzene, contact sensitized guinea pigs show increased DNA synthesis in vitro when stimulated by dinitrophenylated macrophages . In this study, macrophage-containing cell suspensions were isolated from various sources (spleen, lungs, peritoneal cavity) and from the peritoneal cavity also after different stimuli (oil, thioglycollate, starch, lymphokines) . These cells were then haptenized and investigated on their capacity to act as stimulator cells in vitro . In addition, a possible relationship between Ia-positivity of the hapten-presenting cell suspensions and the induction of DNA synthesis was studied . The results demonstrate (1) that the hapten-presenting capacity of macrophages differs with respect to the source and the way of elicitation, (2) that oil-induced peritoneal exudate cells are the most suitable stimulator cells for in vitro assays in contact sensitivity, and (3) that the cellular expression of Ia antigens is in itself no warrant for hapten-presenting capacity. Carcinogenesis, 1982, 3(11), 1317 - 20 Benzo{a}pyrene--DNA adduct formation in target cells in a cell-mediated mutation assay; Sebti SM et al.; In order to analyze the adducts formed in V79 Chinese hamster target cells in a Syrian hamster embryo (HE) cell-mediated mutation assay, a procedure was developed in which the HE cells are differentially killed when the mixed cell suspension is treated with antiserum to Syrian HE cells and complement . The V79 cell suspension, separated from lysed HE cells by gradient centrifugation, is greater than 90% HE cell-free . When the carcinogen--DNA interaction products formed in these two cell types were analyzed after exposure to {3H}benzo{a}pyrene (B{a}P) for 24 h under the conditions of a cell-mediated mutation assay, the major B{a}P--DNA adducts in both cell types resulted from reaction of the anti and syn isomers of B{a}P-7,8-diol-9,10-epoxide with DNA . The amount of B{a}P bound/mg DNA in the target cells was only 30% less than in the activator cells, and the relative proportions of the adducts of the syn and anti isomers were similar in the two cell types . The target cells, which do not metabolize B{a}P, were also unable to metabolically activate B{a}P-7,8-diol to DNA-binding metabolites . Thus, the transfer of activated B{a}P metabolites from activator cells to the DNA of target cells is a relatively efficient process that appears to be independent of the relative reactivity of specific metabolites with cellular nucleophiles . The immunological cell separation procedure we describe can be adapted to the analysis of carcinogen--cell interaction products formed in cell-mediated assays in which other types of activator and target cells are used to measure either mutation or another biological endpoint. Acta Haematol, 1982, 68(4), 309 - 12 Technique for human bone marrow harvest; Herrmann RP et al.; The usual technique of harvesting bone marrow for allogeneic or autologous transplantation involves passage of the marrow suspension through discs of stainless steel mesh of increasingly small diameter . We describe a sterile technique which is much less messy and produces a single cell suspension . Potter-Elvehjem homogenizers are used to break up the marrow particles . This procedure has been used successfully in 6 patients where allogeneic transplantation was performed and in 6 harvests of autologous marrow . Marrow cryopreserved in this way contains viable committed stem cells and is not subject to agglutination following subsequent thawing. Stem Cells, 1982, 1(4-5), 240 - 9 A two-step procedure for obtaining 80-fold enriched suspensions of murine pluripotent hemopoietic stem cells; Visser JW et al.; A method to enrich for murine pluripotent hemopoietic stem cells is described . It consists of two steps . Cells with a density of less than 1.072 g/cm3 at 4 degrees C are first selected from mouse bone marrow by a single-step bovine serum albumin density separation . These low-density cells are then labeled with fluoresceinated wheat germ lectin and sorted by a light-activated cell sorter on the basis of differences in light scatter and fluorescence intensities . Numbers of pluripotent stem cells were determined by the spleen colony assaY (CFU-S) . The concentration of CFU-S was 80 times higher in the sorted cell suspension than in the unfractionated control. Vet Med (Praha), 1982, 27(6), 371 - 7 {Determination of the gonadotropin activity of biological preparations by testosterone synthesis in mouse testes in vitro}; Mamode MI et al.; The ability of mouse testicular tissue to respond to stimulation by a commercial preparation of human choriongonadotropin (Praedyn Spofa) was tested in vitro, using whole testes, fragments and cell suspension . In each experiment seven gonadotropin concentrations ranging from 0.06 to 50 i . u . per litre were tested, and the results were compared with the group of control samples without gonadotropin . The incubation of whole testes with gonadotropin of different concentrations was performed in five vessels, each containing one testis; the fragments were incubated in four vessels, each containing four fragments from different testes, and the cell suspension in triplicates containing 1 . 10(6) cells per ml . All three types of tissue reacted to the increasing concentrations of stimulating agent by the increasing production of testosterone, the concentration of which was determined by radioimmunoassay . For the suppression of the individuality and for achieving a uniform response of the individual samples with each stimulation, the cell suspension of testes from several animals appears to be most suitable. Biorheology, 1982, 19(1/2), 253 - 67 Flows of red blood cell suspensions through narrow two-dimensional channels; Chan T et al.; Apparent viscosity and mean channel hematocrits have been measured at various shear rates and feed hematocrits for red blood cell (RBC) suspensions flowing in two-dimensional channels . Three types of RBC were used in the suspensions : normal, partially hardened by heating at 50 degrees C and completely hardened by glutaraldehyde fixation . Channel height was varied from 20 to 200 mu and feed hematocrit from 5 to 55% . Measurements show that RBC deformability plays a dominant role in narrow channels and viscosity increases rapidly with decreasing cell deformability . Like in narrow tubes the apparent viscosity as well as the mean channel hematocrits decrease as the channel height is reduced . However the apparent viscosity in a channel remains slightly higher than the viscosity in a tube of diameter equal to the channel height . These results are consistent with the existence of a cell-depleted layer near the channel walls. Prostaglandins, 1982 Jan, 23(1), 17 - 28 The in vitro production of prostanoids by cultured bovine articular chondrocytes; Mitrovic D et al.; Bovine articular chondrocytes, cultured as cell suspensions and monolayers, produced prostaglandin (PG) E2 and PGI2 (assayed as 6 keto PGF1 alpha), rather less PGF2 alpha and irregular quantities of thromboxane (Tx) B2 . Addition of foetal calf serum to the medium greatly stimulated PG production (a sixfold increase in PGE2 and a twofold increase in 6 keto PGF1 alpha) . Prostanoid production by cell suspensions grown in serum-free medium generally plateaued after 24 hours . In the presence of 20% foetal calf serum, prostanoid production in long-term monolayer cultures increased during the first 6 days of culture . Levels of PGE2 and TxB2 then decreased, while 6 keto PGF1 alpha levels remained high . Indomethacin (10(-6)M) inhibited chondrocyte PG production both in the presence and absence of added arachidonic acid (10(-4)M) . Prostanoids produced by chondrocytes may play a role in the modulation of cartilage metabolism in vivo. Acta Biol Med Ger, 1982, 41(12), 1139 - 43 Flow-cytometric analysis of glucose induced changes of the transmembrane potential and optical density in pancreatic islet cell suspensions; Zuhlke H et al.; Glucose-induced insulin release of beta cells of pancreatic islets is associated with islet cell membrane electrical activity . We measured the membrane potentials of neonatal rat islet cell suspensions by means of "optical probes" by flow cytometry . Moreover, we determined the narrow angle light scattering and light absorption in direction of the laser beam, respectively . In both these cases two or three clusters of the whole cell population showing different responses to glucose could be distinguished . Glucose-dependent depolarization as well as loss in optical density of cells of pancreatic islets should be associated with the physiological function of insulin producing beta-cells . It is concluded that changes of the membrane potential and/or optical density of islet cells produced by insulin secretagogues can be analysed by flow cytometry which might provide the basis for sorter experiments to separate specific subpopulations of cells of pancreatic islets. Int Arch Allergy Appl Immunol, 1982, 69(3), 210 - 17 Characterization of human lysozyme (muramidase)-releasing cells . Organ distribution and functional properties as analyzed in a protein A plaque assay; Smith CI et al.; Using a modification of the protein A plaque assay, human lysozyme-releasing cells were detected as plaque-forming cells (PFC) . Blood and bone marrow contained higher numbers of muramidase secretors compared to adenoid and tonsil cell suspensions . Human peripheral blood cell cultures were found to contain cells detected as PFC in the presence of antimuramidase immunoglobulin as a developing agent . Lysozyme-producing cells were also found in human bone marrow cultures . High cell densities and a short culture period were optimal conditions for lysozyme release in bone marrow cell cultures . Addition of the activating ligand lipopolysaccharide directly into the plaque assay altered the number of muramidase-secreting cells. Cell Tissue Res, 1982, 225(1), 65 - 77 Reformation of organized epidermal structure by transplantation of suspensions and cultures of epidermal and dermal cells; Worst PK et al.; The development of epidermis and epidermal appendages from dissociated cells of neonate mouse skin was examined by transplantation of cell suspensions to subdermally prepared, protected graft beds . Using Ficol gradients and culture procedures, we prepared subfractions of primary cell suspensions consisting of essentially pure epidermal cells or fibroblasts . Reformation of an epithelium structurally similar to the epidermis was observed from transplanted epidermal-cell suspensions, but formation of hair follicles and development of normal epidermal microarchitecture was observed only when epidermal cells were transplanted together with cells of dermal origin . This pattern was observed following transplantation of either fresh-cell isolates or cells cultured up to 7 days prior to transplantation. Acta Neurochir (Wien), 1982, 65(1-2), 15 - 27 In-vitro secretion of ACTH in Nelson's syndrome; Ludecke DK et al.; Cell suspensions of ACTH cell adenomas of 10 patients with Nelson's syndrome were investigated for in-vitro secretion of ACTH . Two incubation systems, one using incubation beakers and the other a superfusion system, were employed . The cells were tested for their reactivity to lysine-vasopressin (LVP), cortisol, and combinations of both . LVP regularly provoked a rapid significant increase of ACTH secretion . The effect of cortisol was heterogeneous . Paradoxical initial stimulatory effects of cortisol were observed . There was a suppressive effect in some patients, which correlates to low proliferative activity in histological evaluation . In both systems a low secretory activity coincided with high proliferative activity in vivo. Vet Med Nauki, 1982, 19(3), 10 - 5 {Extraction and freeze-drying of herpes virus C3-1 isolated from turkeys}; Kasabov R et al.; Possibilities for producing dried preparations of turkey herpes virus C3-1 by use of various ultra-sound desintegrator-systems and different ways of processing the cell suspensions containing the virus were studied . It was established that cell-less virus of a 1.5 to 2 log smaller quantity than the initial virus containing cell suspension can be produced by ultrasound generator with an indirect action -- a vibrating membrane, which transmits the impulses through a liquid medium and the container's walls on the object . The use of a generator with direct action -- titanic probe placed into the suspension results in an insignificant loss and in some cases even leads to higher titer . In the process of freeze drying the protective media SPGA, 5% pepton and 5% horse serum produce almost equal results . Highest dried virus yield was produced by lyophilization of entire (without centrifugal separation) ultrasounded cell suspension . In such cases the titer of the freeze-drying virus was by 0.2-0.4 log higher than the initial cell virus determined after the microplate method . The authors are of the opinion that titration by this method does not give an exact criterion for comparison of virus content in frozen cell and freeze-dried preparations . A proper protection against Marek's disease was established in laboratory and terrain experiments with chickens following immunization by freze-dried preparations of the strain C3-1 (losses were reduced 3-4 times) . An immunization dose of 2000 POU lyophilized virus is recommended for practical use. Cell Tissue Res, 1982, 223(1), 73 - 86 Dibutyrylic cyclic AMP and theophylline inhibit proliferation of accessory cells in primary cultures of adrenomedullary cells; Unsicker K et al.; Studies on isolated adrenal chromaffin cells in primary cultures may be seriously hampered by the presence of non-chromaffin, mainly fibroblast-like cells, which always occur in dissociates of adrenal medullary tissue and often outnumber the chromaffin cells by the end of the first week of culture, when no measures are taken to control their proliferation . The present study offers a new means to inhibit effectively the proliferation of these accessory cells by treating the cultures with dibutyrylic cyclic AMP (dbcAMP, 0.1 or 0.01 mM) and equimolar amounts of the phosphodiesterase inhibitor theophylline . With this treatment cultures of young rat adrenal chromaffin cells remain virtually free of accessory cells for two weeks of culture . Cultures of bovine adrenomedullary cells retain their initial amounts of non-chromaffin cells, which largely depends upon whether the primary cell suspensions have undergone differential plating prior to seeding . Suppression of accessory cell proliferation with dbcAMP and theophylline is partly due to maintaining differentiation of cortical cells, which otherwise dedifferentiate into rapidly dividing fibroblast-like elements . However, a more direct action of dbcAMP on accessory cells in terms of growth control is also conceivable . DbcAMP and theophylline in the doses applied do not impair the viability, ultrastructure and catecholamine-storing capacity of cultured chromaffin cells. J Reticuloendothel Soc, 1982 Jan, 31(1), 17 - 30 Release of erythropoietin from macrophages mediated by phagocytosis of crystalline silica; Rich IN et al.; The hormone erythropoietin (Ep), which regulates erythropoiesis, can be shown to be released extrarenally from macrophages of the spleen, bone marrow, peritoneal exudate, lung, liver, and fetal liver when suspensions of these cells are preincubated with crystalline silica . The cell supernatants are assayed for Ep activity in the erythroid colony-forming technique using 12- 13-day fresh or cryopreserved fetal liver cell suspensions as a source of Ep-sensitive CFU-E target cells . This report describes the optimal conditions required for release of Ep from silica-treated spleen macrophages . The requirement for phagocytosis of silica in mediating Ep release is shown by four different methods, namely, temperature dependency, inhibition of phagocytosis by Cytochalasin B, the presence of calcium in the medium, and the fact that silica particles coated with serum or poly-2-vinylpyridine-N-oxide can decrease Ep release to control levels . In addition, the reduction to background Ep levels when reduced glutathione, alpha tocopherol (vitamin E), and alpha-thioglycerol are added either separately or in combination with silica-treated macrophages is discussed in terms of a possible mechanism of silica action. J Lab Clin Med, 1982 Jan, 99(1), 56 - 63 Contrasting patterns of protein kinase activities among normal peripheral blood lymphocyte subpopulations and lymphoid cell lines; Elias L et al.; Human peripheral mononuclear cell suspensions were separated into E-rosette-positive (T cell) and E-rosette-negative (non-T cell) fractions and assayed for cAMP binding activity and histone and casein kinase activity, in the presence and absence of cAMP . Each of these activities was higher in E-rosette-positive than E-rosette-negative fractions . Cyclic AMP binding and histone kinase activities were further noted to be higher in T lymphoblastoid cell lines when compared to lines of non-T lymphoid cell origin . In addition . DEAE-cellulose chromatography revealed a qualitative contrast of isoenzymic composition of protein kinase from these cell lines . Cyclic AMP-independent casein kinase activity was high in both types of cell lines . These results suggest that high protein kinase activity is characteristic of T cells and may be related to their higher sensitivity to cAMP agonists in comparison to non-T cells . This study also provides further evidence for an association between increased cAMP-independent protein kinase activity and the transformed state, which has been noted for a variety of other cell types. Gastroenterology, 1982 Jan, 82(1), 106 - 10 Production of angiotensin I converting enzyme by liver granuloma macrophages of Schistosoma mansoni infected mice; Weinstock JV et al.; Angiotensin I converting enzyme activity is detectable in serum and isolated liver granulomas of mice infected with Schistosoma mansoni . At the chronic phase of the infection (20 wk) the intensity of the granulomatous response spontaneously diminishes . Concomitantly, an increase in angiotensin I converting enzyme activity is observed . The objective of this investigation was to determine the source of this elevated angiotensin I converting enzyme activity . In chronically infected mice, measurements showed that angiotensin I converting enzyme activity in hepatic venous blood was higher than that of peripheral venous blood . In contrast, normal mice demonstrated no difference in regional venous angiotensin I converting enzyme activity . Isolated liver granulomas cultured in vitro released angiotensin I converting enzyme into the culture medium . When granulomas were dispersed into single cell suspension, angiotensin I converting enzyme activity was traced to the adherent cell fraction . Both granulomas and adherent cells from granulomas of the chronically (20 wk) infected mice secreted far more angiotensin I converting enzyme than those from animals with acute (8 wk) infection . Cycloheximide partially inhibited enzyme release . We conclude that in murine schistosomiasis, angiotensin I converting enzyme is produced by the granuloma macrophage . The enzyme is released from the granuloma into the circulation probably resulting in increased serum angiotensin I converting enzyme activity . The role of elevated angiotensin I converting enzyme activity within the lesion as well as the circulation remains to be elucidated. Cancer Immunol Immunother, 1982, 13(3), 190 - 3 Cytostatic activity on tumour cells of monocytes from patients with gastrointestinal cancer; Mytar B et al.; The ability of monocytes from patients with gastrointestinal cancer to inhibit tumour cell growth and suppress PHA-induced lymphocyte response in vitro was assessed . Isolated monocytes, i.e., adherent Fc+ cells from mononuclear cell suspension, were cytostatic but not cytolytic for both K562 line and L1210 lymphoma cells . Monocytes from the patients showed an increased ability to inhibit the growth of L1210 but not K562 line cells . The increased cytostatic activity of monocytes was associated with their suppressor activity . This suggests that suppressor monocytes are also able to arrest tumour cell growth in vitro. Acta Biol Med Ger, 1982, 41(12), 1145 - 50 Pancreatic islet cell suspensions of newborn rats and the formation of pseudo-islets in culture; Schroder D et al.; Well preserved pancreatic islet cell suspensions were isolated by trypsin treatment from islets of Langerhans of newborn rats . (Pro)insulin biosynthesis of freshly isolated islet cells can be stimulated by increasing glucose concentrations . In contrast insulin secretion on glucose as the only stimulus is drastically reduced but can be potentiated by 3-isobutyl-1-methylxanthine (IBMX) . Within a period of 4-6 days in tissue culture pancreatic islet cells of newborn rats show the tendency to aggregate and form pseudo-islets of different size. Acta Derm Venereol, 1982, 62(5), 377 - 81 Identification of mononuclear cells in cell suspension from cutaneous infiltrates freed by a suction blister method; Ahokas T et al.; Mononuclear cells were liberated from superficial skin inflammations by a suction blister method which yielded a sufficient number of cells for histochemical and immunocytochemical analysis . The cell population consisted mainly of T cells which were identified as sheep red blood cell rosettes . Very few B cells were found as rosettes with immunobeads . Macrophages and eosinophils were found in varying numbers and they were identified histochemically . The method is suitable for studying nontumorous skin infiltrates. Folia Haematol Int Mag Klin Morphol Blutforsch, 1982, 109(2), 290 - 306 {Separation of red blood cells from G6PD-deficient patients in dextran density gradients}; Grieger M et al.; Red blood cells of 30 patients with G6PD deficiency were separated and characterized by means of isopyknic dextran density gradient centrifugation . The simultaneous determination of G6PD activity and the percentage of NADPH deficiency cells in relation to the maturation parameters of density, reticulocyte share, GOT and PK activity made it possible to recognize differences in the maturation of red blood cells with G6PD deficiency in normal persons as well as within a group of patients . In each case the more or less diminished enzyme activity of the cell suspension was accompanied by a marked enzyme deficiency of the youngest fraction . It is possible that NADPH defect cells are being eliminated at first . In many cases a direct correlation between the percentage of "empty cells" and the in vitro stability tests with and without NADP+ addition could be identified . Decreased maximal speed, changed kinetic behaviour, and instability of these variants are stressed as being the decisive parameters for the life expectation of red blood cells in patients with G6PD deficiency. Virchows Arch A Pathol Anat Histol, 1982, 395(1), 87 - 98 Intracellular keratins in normal and pathological bronchial mucosa . Immunocytochemical studies on biopsies and cell suspensions; Bejui-Thivolet F et al.; The distribution of intracellular keratins was investigated in normal bronchial epithelium and in several morphologically distinct forms of respiratory tract carcinomas . This study was performed with two different experimentally produced antisera against normal human stratum corneum keratin and against keratin protein of MW 67,000 dalton, using indirect immunofluorescence and immunoperoxidase methods on tissue sections and cell suspensions . In normal bronchial epithelium, the basal cells were strongly labelled by both antisera . The ciliated columnar cells appeared devoid of cytokeratins in tissue sections but were strongly labelled with both antisera in cell suspensions . The goblet cells remained negative in every case . In squamous metaplasia of the bronchus, all epithelial cells were unevenly stained with both antisera . Among tumours, only the squamous cell carcinomas were strongly labelled by both antisera . Primary lung adenocarcinoma appeared weakly positive, whereas metastatic lung carcinomas, undifferentiated lung carcinomas, oat cell tumours, carcinoid tumours were negative . The immunocytochemical determination of keratins appeared to be of value in the study of normal and abnormal epithelial differentiation, in the diagnosis of poorly differentiated carcinomas and in their distinction from metastatic tumours of the lung. Scand J Immunol, 1982 Jan, 15(1), 55 - 61 Studies on human epidermal langerhans cells . IV . HLA-D restriction of langerhans cell-dependent antigen activation of T lymphocytes; Braathen LR et al.; Epidermal cell suspensions were produced containing 3-6% HLA-DR-positive Langerhans cells . By using mixtures of epidermal cells and allogeneic T lymphocytes from donors sensitized to purified derivative of tuberculin (PPD) and herpes simplex virus (HSV), we could show that a proliferative in vitro response to these antigens, similar in strength to that seen in autologous combination of macrophages or epidermal cells and T cells, could be obtained provided the epidermal cells shared both HLA-D/DR determinants with the T-cell donor . No antigen-specific response qas obtained in fully HLA-D/DR-disparate combinations . These results, together with previously reported data using other antigen-presenting cells, further emphasize that one minimal requirement for antigen-specific activation of T cells is co-recognition of products of the self HLA-D region. Br J Cancer, 1982 Jan, 45(1), 61 - 9 Lymphoreticular cells in human brain tumours and in normal brain; Phillips JP et al.; The present investigation, using various rosetting assays of cell suspensions prepared by mechanical disaggregation or collagenase digestion, demonstrated lymphoreticular cells in human normal brain (cerebral cortex and cerebellum) and in malignant brain tumours . The study revealed T and B lymphocytes and their subsets (bearing receptors for Fc(IgG) and C3) in 5/14 glioma suspensions, comprising less than 15% of the cell population . Between 20-60% of cells in tumour suspensions morphologically resembled macrophages and less than or equal to 75% of these cells formed strong rosettes . Lymphocytes were not found in cancer-free (putatively normal) brain . Macrophages and the smaller "microglial cells" (both phagocytic, staining with sudan black, and expressing Fc(IgG) and C3 receptors) were found in normal brain in numbers similar to those in tumour suspensions, but with less rosetting avidity . These cells may be part of an immunological defence mechanism. Int Arch Allergy Appl Immunol, 1982, 67(1), 44 - 8 Evidence that tissue mast cells derive from mononuclear phagocytes; Czarnetzki BM et al.; A new method has been developed that allows in vitro differentiation of rat mast cells from precursor cells in the absence of feeder layers . In the present investigation, the precursor cells are further characterized as to their nature, state of activation and distribution in different organs of the rat . The data support our previously published evidence, based on morphology and enzyme-cytochemical reaction patterns that the precursor cells are mononuclear phagocytes . Peritoneal macrophages elicited by mineral oil, thioglycollate or proteose-peptone have lost the ability to differentiate into mast cells . Single cell suspensions of spleen and bone marrow contain mast cell precursors, while cells from mesenteric lymph nodes and thymus or purified lymphocytes never yield mast cells in this culture system. Virchows Arch B Cell Pathol Incl Mol Pathol, 1982, 41(3), 253 - 65 Histochemical study on human germinal centre, mantle-zone and extra-follicular area lymphoid cell subpopulations . Immunological and cytochemical correlations with lymphomatous cells, peripheral normal and leukemic lymphocytes; Palestro G et al.; Tissue sections of lymph nodes, appendices and tonsils, together with smears of immunologically separated peripheral lymphoid cells from a B-CLL and lymphomatous cells from an immunocytoma were submitted to combined enzyme cytochemical investigations with acid alpha-naphthyl acetate esterase (ANAE), beta-glucuronidase (B-G), acid phosphatase (AcPh), adenosine triphosphatase (ATPase), a,d 5'nucleotidase (5'N) . T-cells were Acph+, ATPase- and 5'N- . The vast majority of T- and B-cells displayed ANAE and B-G activities with two distinct staining patterns (T-like and B-like pattern) . A high proportion of lymphoid cells in the germinal centre (G.C.) and the vast majority of lymphoid cells in the mantle-zone (M.Z.) were shown to belong to B-cell system because of the expression of ATPase and 5'N in their membranes . Some lymphoid cells positive for ANAE and B-G with a B-like pattern and for AcPh were recognizable in the G.C . In the M.Z . only a few lymphoid cells being ANAE+, with a T-like pattern, and AcPh+ were shown to belong to the T-cell system . In contrast, in this zone a high proportion of small lymphoid cells (64% +/- 10%) showed ANAE activity, mostly with B-like pattern . Therefore, these findings indicate that in the M.Z . a high proportion of B-cells ATPase+ and 5'N+ also display ANAE activity . By comparison of the results obtained from lymphoid tissue sections, B-CLL and immunocytoma cell suspensions and normal circulating lymphocytes we can conclude that B-ANAE-positive cells of the M.Z . do not usually appear in the peripheral blood . They circulate in large numbers only in some pathological conditions (like our reported B-CLL) . Therefore, B-ANAE-positive lymphoid cells of the mantle, with a B-like staining pattern, include a wide range of subsets which exclude large lymphoid cells and plasma cells. Toxicology, 1982, 24(1), 33 - 43 Inhibition of hepatic lipogenesis by salicylate; Beynen AC et al.; Salicylate has been found to be an inhibitor of fatty acid synthesis in isolated rat hepatocytes . Half-maximal inhibition of fatty acid synthesis occurs at approximately 2 mM . The inhibitory effect of salicylate on fatty acid synthesis is not relieve by the addition of acetate, suggesting that salicylate inhibits the conversion of acetate into fatty acids . Acetyl-CoA carboxylase activity in homogenates of hepatocytes is not influenced by previous exposure of the intact cells to salicylate . Partially purified acetyl-CoA carboxylase, isolated and assayed in the absence of citrate, is markedly inhibited by salicylate . However, in the presence of 0.5 mM citrate, which is the concentration of this metabolite in the cytosol of the liver cell, salicylate activates the enzyme . Upon treatment of acetyl-CoA carboxylase with salicylate (in the absence or presence of citrate), followed by separation of enzyme and effector on a Sephadex G-25 column, the enzyme activity is enhanced as compared to the salicylate-free control, demonstrating that the inhibitory effect of salicylate (in the absence of citrate) is reversible, but not the stimulatory effect (in the presence of citrate) . Salicylate inhibition of fatty acid synthesis by hepatocytes is not rapidly reversible; hepatocytes preincubated with salicylate followed by a wash procedure (centrifugation and resuspension) still show depressed rates of fatty acid synthesis from acetate upon further incubation . Salicylate was found to prevent pyruvate accumulation in hepatocyte suspensions observed in the absence of this compound; salicylate even induces the disappearance of pyruvate and lactate initially present in the cell suspension . This suggests that salicylate activates pyruvate and lactate consumption, which is most likely related to the well-known fact that salicylate uncouples oxidative phosphorylation . The latter action of the drug will stimulate citric acid-cycle activity . This causes an inhibition of fatty acid and cholesterol synthesis since acetyl units will be specifically channelled into the citric acid cycle and not into the lipogenic pathway . It is concluded that part of the inhibitory effect of salicylate on fatty acid biosynthesis is exerted at (a) step(s) in the conversion of acetate into fatty acids, acetyl-CoA carboxylase not being a target of this compound . In addition, salicylate prevents that pyruvate, generated by glycolysis, enters the lipogenic pathway . The latter effect of salicylate would also explain the observed inhibition of cholesterol synthesis by this compound. Virchows Arch B Cell Pathol Incl Mol Pathol, 1982, 39(1), 87 - 99 Observations on induced carcinosis peritonei of the Guinea pig; van de Molengraft F et al.; A diethylnitrosamine-induced hepatocellular carcinoma cell suspension (line-10), injected intraperitoneally in Sewall Wright strain-2 guinea pigs, causes ascites with implantation of malignant cells on the peritoneal surface . At these sites, swelling of the mesothelial cells and simultaneous proliferation of underlying fibroblasts and capillaries are seen . When there are about four layers of malignant cells and the mesothelial lining is disrupted, papillary projections of fibroblasts with capillaries, covered by malignant cells develop . These begin to behave as a "tissue" . In these areas basement membrane destruction and lymphatic and blood vessel infiltration are demonstrable . These developments have been investigated by light microscopy, histochemistry, transmission- and scanning electron microscopy. Mol Immunol, 1982 Jan, 19(1), 39 - 43 The influence of tissue transglutaminase on the function of Fc receptors; Fesus L et al.; In contrast to FcRII the soluble Fc receptor (FcRI) of human peripheral mononuclear blood cells (PMBC) is shed from PMBC following a 4-37 degrees C temperature shift and inhibits rosette formation of nonshed PMBC with antibody-coated erythrocytes (EA) . Purified FcR, could be polymerized by tissue transglutaminase as was revealed by SDS-polyacrylamide gel electrophoresis . Comparing the Sephadex G-150 elution profile of the EA rosette inhibitory capacity of FcRI vs FcRI incubated in the presence of transglutaminase, the latter was found in a higher mol . wt region and could inhibit rosette formation by both FcRI and FcRII . Furthermore, the shedding of FcRI could be prevented by the addition of transglutaminase or Ca2+-ionophore A23187 (which leads to the activation of PMBC transglutaminase) to the cell suspension . The function of FcRII was not affected by either the addition of transglutaminase or Ca2+-ionophore to the cells . The results point to the involvement of transglutaminase in the determination of the functional state of the Fc receptor on the cell surface. Boll Ist Sieroter Milan, 1982, 61(3), 218 - 24 Interferon production in mixed cultures of murine leukocytes and syngeneic L1210 leukemia cells; Sarzotti M et al.; Spl