Hum Genet, 1980, 53(3), 327 - 33 {Preliminary study of stages of human meiosis in spermatocytes and ovocytes (author's transl)}; Geneix A et al.; Human meiotic chromosomes, from spermatocytes and ovocytes, are described after observations of whole mount preparations under E.M . Small testicular and ovarian fragments are put in distillated water, then macerated; the cell suspension is spread on the surface of sheet copper grids covered with formvar plus collodion films . After dehydratation interesting stages are selected under L.M . before observations under E.M . Zygotene and pachytene are the most common stages . During pachytene the chromomeres are well individualized; the synaptonemal complex may be observed; chromatin fibers connect the chromosomes to nuclear pores, interchromosomal fibers joint the bivalents . Zygotene and pachytene bivalents are very similar in the male and the feminine germ cells. Biofizika, 1980 Jan-Feb, 25(1), 178 - 80 {Nature of the membrane potential of liver cells}; Malenkov AG et al.; Light scattering (90 degrees) of Ehrlich ascites tumor and sarcoma 37 cell suspension in the temperature range 25,60 degrees C was studied (heating velocity was 3 grad/min) . It is found that the scattering curve has the peak at 46 degrees C and two plots which are typical of phase transitions in the membranes; the first plot in the range 40-46 degrees C is reversible and the second one at 46-51 degrees C is irreversible . It is proposed that 46 degrees is a critical temperature for the membranes structure stability and viability of the studied cells. Biofizika, 1980 Jan-Feb, 25(1), 177 - 8 {Temperature phase transitions in cancer cells}; Popov GA et al.; Light scattering (90 degrees) of Ehrlich ascites tumor and sarcoma 37 cell suspension in the temperature range 25-60 degrees C was studied (heating velocity was 3 grad/min) . It is found that the scattering curve has the peak at 46 degrees C and two plots which are typical of phase transitions in the membranes; the first plot in the range 40-46 degrees C is reversible and the second one at 46-51 degrees C is irreversible . It is proposed that 46 degrees is a critical temperature for the membranes structure stability and viability of the studied cells. Eur J Biochem, 1980 Jan, 103(2), 323 - 30 Messenger RNA coding for phenylalanine ammonia-lyase . Characterization and partial purification from cell suspension cultures of Petroselinum hortense; Ragg H et al.; The mRNA coding for phenylalanine ammonia-lyase was partially purified from irradiated cell suspension cultures of parsley (Petroselinum hortense) . The product of cell-free translation of the mRNA in a reticulocyte lysate was isolated by immunoprecipitation and compared with the native enzyme subunit . Evidence for the identity, or at least a great similarity, of both was provided by tryptic-peptide and gel-electrophoretic analyses . Under partially denaturing conditions, phenylalanine ammonia-lyase mRNA sedimented as a 20--21-S molecule in a sucrose gradient and had an apparent molecular weight of about 1.05 x 10(6) on a polyacrylamide gel . Approximately two-thirds of the polynucleotide sequence of the mRNA were estimated to be required as coding sequence for the enzyme . We suggest that phenylalanine ammonia-lyase mRNA is unlikely to code for more than of three coordinately induced enzymes. J Surg Oncol, 1980, 13(1), 39 - 44 Mouse colostomy model for studies on large bowel cancer; Volenec FJ et al.; The development of appropriate animal model systems has been proposed as a means of facilitating the study of human colorectal cancer . This report describes the development and use of a blind-colorectal pouch in a carcinoma mouse model . The blind-pouch was prepared in C57BL/6J mice by surgically exteriorizing the descending colon and producing two stomata in the abdominal wall . The proximal stoma served as an end colostomy and the distal stoma created as a mucous fistula . Surgical closure of the anus thus provided a colorectal pouch . In pilot studies it was found that N-methyl-N-nitrosourea (MNU) and radiation together but neither separately produced tumors in the pouch of surviving mice . Further, inoculation of C38 syngeneic tumor cell suspension into the pouch and immediate closure resulted in tumor takes within three to four weeks . The use of this model in carcinogenesis and the immunology of colon cancer simulate the human colorectal cancer problem more closely than previous animal models. Paroi Arterielle, 1980, 6(4), 207 - 11 {Expression of results in studies of vascular biochemical components during the development of the SHR rat}; Bucher B et al.; Hypertrophy and hyperplasia induced by age and hypertension as well as tissue heterogeneity makes it difficult to analyse biochemical datas obtained with rats thoracic aortas from rats of various age and blood pressure . In SHR, the thoracic aorta weight increases parallely to the body weight from the first week after birth onwards . Meanwhile the protein content of the organ increases parallely to the organ weight . However, though the DNA content increases markedly up to 7 weeks of age, one observes a much lower increase with age afterwards . It seemed to us that the aorta DNA content was more representative of the organ cell number . This led us to use express our biochemical data per mg of the organ DNA content . Moreover, it seems necessary to take in account the cell volume which increases as well with age and hypertension on the one hand, and to be able to distinguish the cellular content of the myocytes from the one of other cells types contained in the adventitia and the intima, on the other hand . For these purposes pure media layers are prepared by completely removing the adventitia and intima and single cell suspensions are obtained after total enzyme digestion of the medias. Beitr Trop Landwirtsch Veterinarmed, 1980, 18(4), 395 - 8 Karyological study of Gallus domesticus macrochromosomes; El-Metainy AY et al.; After an incubation period of four days and colchicine pretreatment (air-cell method), clear metaphase figures were obtained from a cell suspension of the allantoic sacs . It was only by shortening the colchicine application to one hour that sufficiently elongated chromosomes suitable for karyological studies could be prepared . The total arm length and the arm length indices are given of the six pairs of macrochromosomes - including the Z chromosome - and of the three largest microchromosomes . Different from the previous findings as reported in the literature, the two largest macrochromosomes are described as submetacentric and not as metacentric . The findings on the seven chromosomes following agree with those given in the literature. Recent Results Cancer Res, 1980, 75, 252 - 9 Erythrocytes and lymphocytes as drug carrier systems: techniques for entrapment of drugs in living cells; Zimmermann U et al.; Mouse thymocytes and erythrocytes are loaded electrically with drugs in isotonic solution . The loaded cells are used for targeting the drugs to specific sites in the organism in order to achieve a controlled drug release in time and space . The field technique used for the loading of the cells is based on the dielectric breakdown of the cell membrane which is observed when cell suspensions are subjected to external field pulses of 2-20 kV/cm for short time intervals (ns to microseconds) . When an apparent membrane potential of about 1 V is reached in response to the external field, the membrane breaks down reversibly . The breakdown of the membrane is associated with a remarkable and reversible permeability increase of the cell membrane . The increase in permeability depends on the strength and the duration of the field pulse. Arch Dermatol Res, 1980, 268(3), 231 - 7 Increased "in vivo" lymphocyte blastogenesis in the peripheral blood of patients with atopic dermatitis and allergic contact dermatitis; Lachapelle JM et al.; The spontaneous 3H-thymidine (3HT) labelling of some lymphocyte subpopulations has been studied in the peripheral blood of five patients with atopic dermatitis and five with widespread allergic contact dermatitis and compared with that in 10 healthy subjects . One hour after addition of 3HT to heparinized blood, lymphocytes were separated and processed with two different rosetting techniques (E-rosette test and Active E-rosette test) . The cell suspensions were cytocentrifugated and autoradiography undertaken . An increased number of 3HT labelled lymphocytes was observed in the peripheral blood of patients with dermatitis as compared to controls . These labelled lymphocytes were E-rosette-forming cells (T cells) and E-non-rosette-forming cells (non-rosette-forming T cells and non-T cells) . The ratio between the labelling index (LI) of E-rosette- to the LI of non-E-rosette-forming cells was in favour of T cells in allergic contact dermatitis (ratio = 3.09) whereas in atopic dermatitis (ratio = 0.93) the DNA synthesis was relatively greater in the non-rosette-forming cells . It is suggested that this increased LI of peripheral blood lymphocytes could be related to the increased derman mononuclear cell 3HT-labelling that has been reported previously in these inflammatory skin diseases. J Immunol Methods, 1980, 35(1-2), 43 - 56 An anti-human t-lymphocyte antiserum . Further characterization of the specificity for T-cell subpopulations and a comparison of methods for identification of T-lymphocytes in tissue sections; Braathen LR; An anti-human T-lymphocyte antiserum has been further studied for specificity for T-cell subpopulations and application in tissue sections . Using a complement-dependent microcytotoxicity assay about 60% of normal peripheral blood T-cells were found to be sensitive to lysis, while 40% were resistant . When the T-cell suspensions were depleted of TG-cells using EA (Ripley) rosette sedimentation, the remaining cells demonstrated an increased percentage of lysis-sensitive cells, while the T-cells enriched in EA-RFC demonstrated a decreased percentage of cells sensitive to lysis, indicating that the antiserum was primarily directed against the non TG-cells, i.e . probably the TM helper cell population . This was further supported by functional studies . In order to quantitate T-cells, cytocentrifuge preparations were made from cell suspensions with known T-cell percentages and the T-cells determined with both the immune adherence technique using AET-treated sheep erythrocytes, as well as the indirect immunofluorescence technique using anti-T antiserum . The results of the two methods correlated well, suggesting that both methods can be used to determine T-cells in situ in infiltrate-like clusters of cells. J Immunol Methods, 1980, 37(3-4), 275 - 86 The intracellular localisation of immunoglobulin in human lymphoid cells and haematopoietic cell lines by immunoperoxidase electron microscopy; Newell DG et al.; A technique is described for the ultrastructural localisation of intracellular immunoglobulin (Ig) in human lymphoid cell suspensions by the immunoperoxidase method . The technique involves restricted saponin digestion of glutaraldehyde prefixed cells to enhance conjugate penetration . With this method of staining Ig was located in the rough endoplasmic reticulum, perinuclear space and Golgi apparatus in human lymphoid cells from a variety of sources, which is consistent with previously published observations using other ultrastructural techniques . In contrast, diffuse intracytoplasmic staining was predominant in cells prefixed with glutaraldehyde but not treated with saponin . These differences in patterns are discussed in terms of membrane permeability . Although saponin treatment was necessary for consistent localisation of intracellular Ig it resulted in unavoidable loss of Ig from the surface of the cells. Folia Microbiol (Praha), 1980, 25(5), 361 - 8 Characterization of adenylate cyclase from Escherichia coli; Janecek J et al.; Adenylate cyclase activity was detected and characterized in cell-free preparations of different strains of Escherichia coli; it was localized not only in the membrane fraction but also in the cytoplasm, the localization differing from strain to strain . The adenylate cyclase activity is highly dependent on the method used for disintegration of cells . The best results were obtained when using vortexing of the cell suspension with ballotini beads . The pH optimum of adenylate cyclase in cell-free preparations was found to be 9.0--9.5 . The enzyme has an absolute requirement for Mg2+ and is inhibited by sodium fluoride and inorganic diphosphate . Release of adenylate cyclase from the membrane leads to an immediate loss of the activity; it was found that adenylate cyclase is quite labile and hence it could not yet been purified . The method used to determine adenylate cyclase activity and cyclic AMP is described. Adv Exp Med Biol, 1980, 132, 509 - 17 Isolation of a lipocyte-rich fraction from rat liver nonparenchymal cells; Otto DA et al.; A modified pronase digestion procedure is described for isolating nonparenchymal liver cells from vitamin A treated rats, which yielded a 15-23% population of lipocytes in the total cell suspension . The criteria for defining a lipocyte was the appearance of vitamin A containing cells as determined by fluorescent microscopy . Measurement of alcohol dehydrogenase and retinol dehydrogenase activities indicated that these enzyme activities were not present in the isolated nonparenchymal cells . A lipocyte-rich fraction of nonparenchymal cells was obtained by centrifugation of purified nonparenchymal cells in a linear Metrizamide gradient . Vitamin A fluorescence and chemical assay of vitamin A in the cell fractions indicated a four-fold enrichment of lipocytes in the cell fraction with d = 1.043 g/ml . Fractions high in vitamin A also had numerous cells with fat droplets as shown by transmission electron microscopy. J Clin Lab Immunol, 1980 Jan, 3(1), 51 - 61 Lymphocyte subpopulations in the thymus of SJL/J mice: age-related alterations and the effect of spontaneous reticulum cell sarcoma development; Dumont F; The cellular composition of the thymus was investigated as a function of age in the immunologically aberrant SJL/J mouse strain . Lymphocyte subpopulations were identified by combined analysis of cell electrophoretic mobility (EPM) and cell electronic volume and by assessment of surface receptor for Peanut-agglutinin (PNA) and of surface immunoglobulin (slg) determinants . In the thymus from young adult animals two major types of thymocytes could thus be recognized . The first one (th1, 2, 3) representing about 75% of the thymocyte population was endowed with a low-EPM and exhibited PNA-receptors . The other one (th4) possessed a high-EPM and lacked PNA-receptors . During ageing of mice selected for the absence of macroscopically detectable Reticulum Cell Sarcoma (RCS) lesions, the frequency of th1, 2, 3 cells diminished whereas that of th4 cells increased (up to 75% at the age of 16 months) . This latter augmentation reflected a true expansion of the th4 cell subpopulation and acounted for the maintenance of thymus cellularity to a relatively high level throughout life . In accordance with the probable immunocompetence of th4 cells, thymus cell suspensions from old RCS-free SJL/J mice were found to exhibit high proliferative responses to T-cell mitogens . On the other hand, from the age of 8 months onwards, a new physical type of lymphocytes (th5) could be detected in increasing proportions . These cells were characterized by a lower EPM than typical th1, 2, 3 thymocytes and by a modal volume around 150 micrometer3 . They were further demonstrated to be PNA- but slg+ and are thus likely to represent B cells . Such alterations were not encountered in BALB/c and DBA/2 mice in which both th1, 2, 3 and th4 thymocyte subpopulations regressed at approximately the same rate with age . Moreover, in the thymus of RCS-bearing SJL/J mice, the hyperplasia of the th4 cell pool and the occurrence of th5 cells appeared less important than in RCS-free mice. J Invest Dermatol, 1980 Jan, 74(1), 54 - 8 The epidermal cell which selectively adheres to a collagen substrate is the basal cell; Stanley JR et al.; In order to determine whether a specific subpopulation of epidermal cells selectively attaches to collagen substrates in vitro, epidermal cell suspensions, obtained by trypsinization of guinea pig skin, were incubated on type I or type IV collagen-coated glass cover slips . It was noted, morphologically and by electronic volume measurements, that small round cells, as opposed to the larger angulated flat cells, adhered to the collagen substrates . To further characterize the attached cells, the percentage of basal cells was determined in the attached cell population and in the initial epidermal cell suspension . Basal cells were identified by indirect immunofluorescence in 2 ways: (1) by the presence of pemphigoid antigen and (2) by the absence of upper cytoplasmic antigen, which is present in all keratinocytes except the basal cells . Whereas in the initial guinea pig epidermal cell suspensions about 50% of the cells were basal cells using either of these 2 criteria, 86-97% of the cells which adhered to the collagen substrates were basal cells . Human basal cells, as defined by pemphigoid antigen, also selectively adhered to the collagen substrates. Scand J Immunol, 1980, 11(4), 401 - 8 Studies on human epidermal Langerhans cells . I . Allo-activating and antigen-presenting capacity; Braathen LR et al.; Human epidermis was separated from dermis by means of a suction blister device and dissociated with trypsin . The epidermal cell suspensions obtained contained 3--5% Langerhans cells as judged by immunofluorescence staining ot the cells with a rabbit anti-DR antiserum . The epidermal cells were co-cultured with purified allogeneic T cells and with autologous T cells with or without PPD of tuberculin . A strong T-cell response to allogeneic epidermal cells was obtained, as was a strong T-cell response to PPD, provided autologous epidermal cells were also present . Pre-treatment of the epidermal cells with anti-DR antiserum plus complement abolished both these responses . These data indicate that epidermal cells are able to substitute for macrophages both in the allo-activating and in the antigen-presenting function . Since the responsible cells were DR-positive, it is highly probable that the cells responsible for these functions are the Langerhans cells. Ann N Y Acad Sci, 1980, 341, 57 - 66 Coupling of ion flows in cell suspension systems; Geck P et al.; A valid test for cotransport between solutes is a demonstration that the degree of coupling between all coupled solute flows concerned, q defined in terms of Irreversible Thermodynamics, is sufficiently close to unity . The usual method to determine q kinetically by pulses and responses of flows can not simply be applied to rheogenic ion flows, as electrical potential difference changes due to the pulses can hardly be avoided . If, however, the ion flows are all electrically silent, changes in electrical potential difference (PD.) should not interfere with the determination of q . This holds for the furosemide-sensitive fluxes of Na+, K+, and Cl- in Ehrlich cells, each of which could be shown to be unaffected by a change in electrical PD and vice versa . Hence the q values could be determined for any pair of the three ion flows concerned and none differed significantly from unity . These results appear to indicate a furosemide-sensitive, electrically silent ternary symport mechanism for Na+, K+, Cl- with the stoichiometry 1:1:2, which is active but does not utilize ATP . It is assumed to function as a very efficient regulator of cellular volume and may be identical with other previously described binary symport systems. Dev Biol Stand, 1980, 46, 67 - 74 A novel rapid and continuous method for the resolution of cell dispersal activities in crude trypsin preparations; Dickerson CH et al.; High speed continuous electrophoretic fractionation of crude commercial trypsin preparations has resulted in the resolution of the cell monolayer dispersal activity into three fractions, two of which were considered useful in the production of fully dispersed single cell suspensions . This separation could be achieved at an input rate for crude enzyme of 500 mg per minute. Acta Biol Med Ger, 1980, 39(11-12), 1165 - 75 {In vitro cultivation and behavior of aortic endothelium cells in a low serum culture medium}; Halle W et al.; Endothelial cells isolated from calf aorta were used in the subculture No . 4 to 10 for experiments to establish standardized and well reproducible conditions of cultivation . The cells can be cultivated in the commercial medium Eagle-MEM with following supplements: 0.1 g L-glutamine, 5.0 g peptone, 0.5 g serum albumin, and 10 ml (= 1%) bovine serum per 1000 ml medium (MEMPAS) . With the aid of immunofluorescence technique the cell type specific marker Factor VIII antigen was shown to be localized especially in the perinuclear region of the cells . The cells were characterized with regard to their growth behaviour . Both, the MEMPAS and the Eagle-MEM with 10 per cent serum increases the cell number in the first 4 days of the exponential growth to the same values . The use of MEMPAS in connection with a strict cultivation regime from the deep frozen cell suspension to culture in scintillation vials guaranteed well reproducible conditions of cultivation . In 14 non-selected experiments distributed over a longer period of time it was found that with regard to the values of the cell number on the respective day the cultures can be divided into two groups, which differ with statistical significance . In further experiments it was possible to confirm this result . Medium, conditioned by endothelial cells (K-MEMPAS) increases the cell number and the growth rate . From these results it was concluded that endothelial cells of vessels are able to produce growth factors with self-stimulating effects . At this time the endothelial cell line is stored in deep frozen state up to the 25th subculture . The endothelial cells cultivated in the described standardized conditions are useful for screening of cell type specific factors with angiogenic activity. Arzneimittelforschung, 1980, 30(7), 1119 - 23 {Nucleic acid and protein synthesis of splenic lymphocytes of the rat under the influence of mucopolysaccharide-polysulfuric acid-esters (author's transl)}; Tempel K; The influence of three mucopolysaccharide-polysulfuric acid-esters (MPS) of different molecular weight on DNAase II-activity as well as on nucleic acid and protein synthesis of lymphocytes of rat spleen has been investigated in vitro . The results are summarized as follows: 1 . DNAase II-activity (bovine spleen) was inhibited by MPS in a competitive manner . 2 . At concentrations of greater than or equal to 10 microgram/ml MPS decreased the incorporation of 3H-thymidine and 14C-uridine into the nucleic acids of the cell suspension . 3 . Incorporation of 3H-amino-acids into the lymphocytes protein was enhanced when MPS were added at concentrations of > 1 and > 10 microgram/ml, resp . At lower concentrations of the polyanions, a slight inhibition of protein synthesis was shown . 4 . In the presence of MPS, incorporation of the radioactively labelled presursors into the acid-soluble fraction of the cells was enhanced . 5 . The MPS-effects described increased with the molecular weight of the polyanions . It is suggested that polyanions like MPS may interfere rather non-specifically with cell membranes and with nucleic acid-polymerases. J Natl Cancer Inst, 1980 Jan, 64(1), 89 - 96 In vivo T-lymphocyte response against spontaneous reticulum cell neoplasms in SJL/J mice; Weinstein Y et al.; The properties of lymphocytes associated with reticulum cell neoplasms (RCN) (type B) occurring spontaneously in SJL/J mice were examined . The activity of 20 alpha-hydroxysteroid dehydrogenase (20 alpha-SDH) was used as a marker for activated T-cells . High levels of this enzyme were found in cell suspensions of tumors taken from 6- to 7-month-old mice . Treatment of the cells derived from tumorous organs with anti-theta serum and complement resulted in a loss of the 20 alpha-SDH activity; this indicated that T-lymphocytes populate the RCN . The activated T-cells in the neoplastic tissue were larger than small lymphocytes . In the more advanced stage of tumor growth seen in 1-year-old mice, the percentage of malignant reticulum cells was low and the neoplastic tissue showed low levels of 20 alpha-SDH activity . Tumor cells irradiated in vitro triggered syngeneic lymphocytes to proliferate in tumor-lymphocyte mixed cultures . The T-cell proliferative response measured by {3H}thymidine incorporation was accompanied by a marked increase in 20 alpha-SDH activity . The spleen cells taken from mice bearing old tumors that showed marked fibrosis did not respond to T- and B-cell mitogens . Histologically, the structure of the spleen was preserved, with few or no tumor cells . Spleen cells from age-matched healthy mice responded to mitogens. Horm Res, 1980, 13(4-5), 259 - 79 In vitro secretion of ACTH, beta-endorphin and beta-lipotropin in Cushing's disease and Nelson's syndrome; Ludecke DK et al.; Tissue from histologically confirmed ACTH cell adenomas in Cushing's disease (CD) and Nelson's syndrome (NS) was gained by transsphenoidal surgery . Combined enzymatic and mechanic agitation of tumor tissue yielded a cell suspension . Aliquots of the cell suspension were transferred to superfusion chambers immediately after isolation and investigated for ACTH and beta-endorphin production . Feedback action of cortisol (CO) and dexamethasone on basal hormone production and on lysine vasopressin (LVP) induced ACTH secretion were studied . Adenomatous tissue and anterior lobe tissue from the same patient in CD could be investigated simultaneously in 4 cases . The paraadenomatous tissue showed depression of basal and LVP-induced ACTH secretion . In all adenomatous tissues investigated there was missing or reduced suppression of basal ACTH secretion by physiological levels of CO . CO not only failed to suppress LVP-induced ACTH secretion but also seemed to enhance LVP stimulation in some experiments . This study confirms former results, that a missing or inversed feedback action or glucocorticoids in adenoma cells is a mechanism involved in the pathological ACTH secretion in CD and NS . Bioassayable and immunoreactive ACTH from media of superfusion and short-term static incubation were compared with beta-endorphin and beta-LPH in an assay detecting these two peptides with equimolar sensitivity . Secretory patterns were basically parallel but great differences showed in quantities of hormones secreted . In addition, Sephadex G-50 gel chromatography was performed to separate beta-endorphin from beta-LPH and to calculate the ratios . These profiles show great variations between different adenomas. Acta Radiol Oncol, 1980, 19(5), 361 - 8 Effects of acute gamma-irradiation on spermatogenesis as revealed by flow cytometry; Hacker U et al.; Mice irradiated with doses ranging from 0.1 to 15 Gy using a 60Co-source and controls were killed at intervals varying from 2 to 35 days after irradiation . The DNA content of the testicular cells in single cell suspensions was measured with the pulse cytophotometer to determine the frequencies of the different stages in the spermatogenesis . The relative amount of S-phase and 4c-cells was reduced initially but increased subsequently to hypernormal values . A decrease of 2c-cells indicated a higher cell-kill of diploid spermatogonia . Gamma ray-induced spermatids with abnormal DNA-values (diploid sperm) were identified . The results suggest that the spermatogenesis can be analysed with flow cytometry and used as a biologic dosimeter even for small doses of ionizing radiation. Physiol Bohemoslov, 1980, 29(2), 107 - 16 In vitro K+-effect on ATP and phosphocreatine levels and on Na+ K+-atpase activity of mouse brain cells; Kovaru H et al.; Cell suspensions were prepared form mouse brain cortices . Cells were incubated in a medium also containing 10 mmol.l-1 glucose and either 5 mmol l-1 K+ or 50 mmol.l-1 K+ concentration . In order to standardize individual experiments, the biuret reaction was modified for rapid determination of the protein in cell suspensions . The cellular reserves of energy-rich phosphates were determined in the course of 60 min of cell incubation with 5 mmol.l-1 K+ and following 30 min incubation of cells with 50 mmol.l-1 K+ . The level of ATP was significantly elevated after 10-60 min of incubation with low K+, from 0.58 to 0.78 micromoles per 100 mg protein; the creatine phosphate content during the same interval was in the range 1.27-1.44 micromoles per 100 mg protein . A significant decrease of energy reserves in cells was observed if the extracellular concentration of K+ was increased . After 10 and 30 min of incubation, a decrease by 36.2% and 38.5% for creatine phosphate and 34.6% and 44.9% for ATP was found, respectively . Na+ K+-ATPase activity of cells incubated for 60 min with 5 mmol.l-1 K+ was expressed as 4.99 micromoles of liberated Pi per 100 mg protein.1 h . Enzyme activity was stimulated with 50 mmol.l-1 K+ by 24.3% and 25.7% after 10 and 30 min of cell incubation respectively . Stimulation of Na+ K+-ATPase activity of brain cortex cells was directly dependent on the actual presence of stimulating 50 mmol.l-1 K+ concentration. Prog Clin Biol Res, 1980, 48, 277 - 90 Further experience in testing the sensitivity of human ovarian carcinoma cells to interferon in an in vitro semisolid agar culture system: comparison of solid and ascitic forms of the tumor; Epstein LB et al.; We studied the in vitro growth characteristics of 10 solid-tumor samples of patients with ovarian carcinoma using a semisolid agar culture technique . Tumor cell colonies were observed in 8 of 10 samples, but sufficient number of tumor colonies to evaluate the effects of interferon and other antitumor agents were obtained in only four samples . As compared with cell suspensions prepared from ascitic fluid samples, solid-tumor samples had markedly lower viability, 39% vs 89%, and had more tumor cells, 81% vs 28% . Also, whereas the maximum increase in tumor-colony number occurred during the first week of growth in both solid- and ascitic-fluid-derived samples grown concurrently from the same donors, increase in tumor colony number was sustained for longer periods in ascitic-fluid-derived cultures . The ascitic-fluid-derived tumor colonies were more sensitive to the antiproliferative effects of interferon than colonies derived from solid-tumors . At a concentration of 300 units/ml incorporated into the agar for the duration of the culture, three of four ascitic fluid samples showed a reduction in tumor colony number by greater than or equal to 25%, whereas none of the solid-tumor samples were affected by the interferon to that degree . In contrast, solid-tumor samples showed greater response to cis platinum and Adriamycin than did ascitic-fluid-derived cultures . Such studies and observations are critical in designing clinical trials for the use of interferon in the treatment of malignancy and the judicious selection of patients and route of administration most likely to provide optimal results, especially in view of present critical shortages in availability of interferon. Acta Derm Venereol, 1980, 60(5), 381 - 7 Studies on human epidermal Langerhans' cells: II . Activation of human T lymphocytes to herpes simplex virus; Braathen LR et al.; Epidermis from patients suffering from recurrent herpes labialis was separated from dermis by means of a suction blister device and dissociated with trypsin . The epidermal cell suspensions obtained were 80--95% viable and contained 3--5% Langerhans' cells, as judged by immunofluorescence staining of the cells with a rabbit anti-DR antiserum . T lymphocytes from the same patients were co-cultured with herpes simplex virus antigen (HSV-Ag) or live virus (HSV), with or without epidermal cells or macrophages . A strong proliferative T cell response to HSV-Ag and HSV was obtained, provided that the cultures also contained epidermal cells or macrophages . Pretreatment of the epidermal cells with a rabbit anti-DR antiserum plus complement abolished the responses, while pretreatment with normal rabbit serum plus complement did not . These data therefore indicate that HLA--DR positive Langerhans' cells are able to present herpes simplex virus in an immunogenic way to T lymphocytes. Med Microbiol Immunol (Berl), 1980, 168(2), 129 - 37 Detection and spontaneous alteration of lymphocyte antigens on slide smears; Sassen A et al.; Smears of cell suspensions from murine lymphoid organs were prepared on slides, air dried, and processed for detection of immunoglobulin (Ig) and theta (Thy-1) antigens by the unlabeled antibody peroxidase-antiperoxidase (PAP) technique . Satisfactory results were obtained for both antigens on recently made smears . However, slides kept at room temperature showed a progressive decrease in staining of the two antigens with time . Various fixatives and preservation procedures were tested to prevent this alteration . Good conservation of smears was obtained when slides were kept at -18 degrees C and/or air isolated before or after fixation with alcohols . A similar degradation of Ig and/or Thy-1 antigens occurred also in histological tissue sections or in serum spots dried on slides . The major cause for this degradation is thought to be contact with air, residual enzymatic hydrolysis playing a less important role. Int Arch Allergy Appl Immunol, 1980, 63(2), 153 - 8 Suppression of experimental allergic encephalomyelitis with thoracic duct lymphocytes; Frost H et al.; Thoracic duct lymphocytes (TDL) from Lewis rats immunized 9-10 days previously with basic protein in complete Freund's adjuvant (BP-CFA) failed to induce experimental allergic encephalomyelitis (EAE) in syngeneic recipients . This contrasts with the successful transfer of EAE by lymph node cell suspensions from donors immunized 9 days previously with BP-CFA . Only minor EAE was induced passively by TDL from rats immunized 11-12 days before with BP-CFA . TDL collected 9-20 days after BP-CFA immunization, however, were successful in transferring specific suppression of EAE tested by the lack of disease in the recipients immunized actively with BP-CFA 1 week after the TDL transfer . The data indicate that the thoracic duct contains specific suppressor cells shortly before, during and after the development of clinical EAE. Folia Haematol Int Mag Klin Morphol Blutforsch, 1980, 107(1), 104 - 24 {Erythrocyte concentrates, low in buffy coat: production, preservation and evaluation of its quality.}; Strauss D et al.; Buffy coat-poor packed red cells were prepared from fresh ACD-, ACD-AG- and EDTA-blood, than resuspended with a preservation solution, containing glucose, adenine, guanosine, sucrose, citric acid and sodium citrate and stored at 4 degrees C for 6 weeks . The survival rate of resuspended red cells from ACD-AG-blood amounted to 77% after 6 weeks of storage . The ATP content of resuspended red cells was approximately 25% lower than in ACD-AG whole blood during storage caused probably by increased ATP consuming reactions at the red cell membrane . The P2G-content of resuspended red cells from ACD- and ACD-AG-blood decreased above 50% of the normal level during the first week, as fast as in ACD- and ACD-AG whole blood . The P2G-breakdown in red cells from EDTA-blood was delayed for a week due to the higher pH as in CPD blood . Additions of xylitol, inorganic phosphate, and bicarbonate in 6, 5 and 20 mM final concentrations in the red cell suspensions and an increased pH at the same time delayed the breakdown of ATP and P2G . Packed red cells can be administered fast enough at hematocrits to 0.60 that will be achieved by adding 50 to 100 ml preservation solution . Leukocytes and thrombocytes were reduceds to 70 to 80% . With increasing rate of reduction a higher loss of red cells occured . Buffy coat-poor red cell concentrate contains only few microaggregates . It diminishes the risc of febrile transfusion reactions and delays the appearance of alloimmunisation . The circulatory overload of patients is less frequent than after transfusions of red cell resuspensions containing a large resuspension volume. Int Arch Allergy Appl Immunol, 1980, 62(2), 205 - 12 Histamine-containing cells from bronchial lavage of macaque monkeys . Time course and inhibition of anaphylactic histamine release; Butchers PR et al.; Bronchial lavage of rhesus and cynomolgus monkeys provided leucocyte suspensions with viable histamine-containing cells (HCC) as 1--8% of the white cell population . These HCC released histamine or challenge with antiserum to human IgE . HCC from 2 monkeys with pulmonary and cutaneous hypersensitivity to Ascaris antigen (AA) released histamine on challenge with AA . The extent of histamine release was related to the concentration of antigen and antiserum, and histamine release was complete within 10 min of challenge . (+/-)Salbutamol and (-)isoprenaline were potent inhibitors of anaphylactic histamine release from HCC, but disodium cromoglycate and AH 9679 were relatively poor inhibitors . The HCC system combines the reproducibility of a cell suspension with a response to drugs similar to that of human lung fragments. Acta Biol, 1980, 31(1-3), 249 - 55 Rate of thymidine incorporation and incidence of parenchymal cell division in adult rat hypophyseal cell cultures . Effect of thyroliberin and somatostatin; Rappay G et al.; Cell suspensions derived from adult rat anterior pituitary glands were cultured for up to eight days . Prolactin immunoreactivity and/or tritiated thymidine incorporation into DNA of cell nuclei were demonstrated in cells with and without thyroliberin (TRH) and somatostatin (SRIF) treatment . It has been established that (a) TRH, which is effective in releasing both thyrotropin and prolactin, may stimulate cell proliferation in other than its target cells; that (b) SRIF has no effect on lactotropic cell proliferation and augments thymidine incorporation into DNA of unidentified cells; that (c) immunoreactive lactotropic cells with tritium-labelled nuclei are present in each culture, independent of hypothalamic hormone treatments. Virchows Arch B Cell Pathol Incl Mol Pathol, 1980, 33(2), 127 - 38 A T-dependent plasma cell response: part of a graft-versus-host and host-versus-graft reaction in the rabbit spleen; Veldman JE et al.; B-cell reactivity, as expressed in a plasma cell reaction may be part of a graft-versus-host or host-versus-graft reaction . Cell transfer experiments were able to prove this . Histological evidence is presented for the occurrence of the so-called "allogeneic effect" during a graft-versus-host or host-versus-graft reaction in the rabbit spleen . Cell suspensions containing both T- and B-cells give rise to a cellular and a humoral immunity reaction component upon transfer to histoincompatible recipients . This can also occur during a graft-versus-host reaction . This B-cell response is T-cell dependent. Virchows Arch B Cell Pathol Incl Mol Pathol, 1980, 33(2), 107 - 16 Diagnostic morphometry of isolated lymph node cells from patients with mycosis fungoides and Sézary's syndrome; van der Loo EM et al.; Mycosis fungoides (MF) and Sezary's syndrome are cutaneous T cell lymphomas, characterized by the presence of lymphoid cells with deeply indented nuclei (CMC) in the infiltrate . In order to find objective criteria for the diagnosis of early MF involvement of lymph nodes from patients with MF, we performed morphometric analysis of lymphoid cells in lymph node cell suspensions measuring the degree of nuclear indentation as expressed by the nuclear contour index (NCI) . Statistical discriminant analysis was used to analyze the differences in the NCI histograms between lymph nodes without and with MF involvement and to select the most discriminating parameters for diagnostic classification . Using a training set of 6 lymph nodes from patients with unrelated diseases and 8 lymph nodes from patients involved by cutaneous T cell lymphomas, the mean and standard deviation of the NCI histograms were selected as the most discriminating parameters . All lymph nodes from the training set were assigned to the correct diagnostic classification group with a probability over 90% . The predictive value of the morphometric classification was tested on a set of 12 enlarged lymph nodes from patients with MF . The histological diagnosis was used as a reference . In 10 cases the morphometric classification was identical to the histological classification, whereas in two cases (1 classified as positive, 1 as negative) a disagreement was found . It is concluded that morphometry of lymphoid cells can contribute substantially to the diagnosis of early MF involvement in lymph nodes. Nature, 1979 Dec 13, 282(5740), 738 - 9 Shear-induced concanavalin A agglutination of human erythrocytes; Greig RG et al.; The mechanism by which cell suspensions are agglutinated by plant lectins remains obscure . The agglutination of a particular cell line in the presence of a specific plant lectin probably depends on several factors including the number and valence of lectin molecules bound to the cell surface, the mobility of receptor molecules in the membrane, the surface morphology and charge and the metabolic state of the cell1-6 . The assay system used to assess cell agglutination also seems to be important, since many laboratories studying the same agglutination reaction have reported dissimilar or contradictory results . To provide further information on the molecular mechanism of agglutination we have begun a systematic study on the aggregation of human red cells by the lectin concanavalin A (Con A) . By using an adaptation of our previously described aggregation assay which provides a continuous measure of both the rate and degree of intercellular adhesion in the presence of controlled shear forces, we have found that the agglutination of erythrocytes by Con A can be resolved into three stages, two of which are observed only if the system is exposed to shear. Eur J Immunol, 1979 Dec, 9(12), 997 - 1003 Surface Ig on rabbit lymphocytes . Rabbit B and T cells are distinct populations; Bast BJ et al.; Rabbit peripheral blood lymphocytes (PBL) were analyzed by immunofluorescence using anti-T cell conjugates and anti-Fab, anti-a1 allotype, anti-IgM and anti-IgA conjugates . In addition, T cells were demonstrated by rosetting with papain-treated homologous erythrocytes . Control experiments, using acid treatment and incubation at 37 degrees C for 18 h after or without pronase treatment, revealed the endogenous origin of all surface determinants tested . A good correlation was found between results obtained with the two anti-T cell conjugates used and the T rosette test on PBL and on lymphoid cells isolated from various organs . In lymphocytes isolated from peripheral blood and from various lymphoid organs, the percentages of T and B cells were respectively 45 and 38 for PBL, 10 and 46 for bone marrow, 27 and 31 for appendix, 40 and 45 for spleen, 42 and 46 for Peyer's patches, 96 and 0.3 for thymus and 70 and 16 for peripheral lymph nodes . The percentage of "null" cells in lymphocyte populations derived from bone marrow and appendix is rather high . The final percentages of T and B cells in rabbit PBL depend to a significant extent on the method of isolation, especially isolation by Ficoll-Hypaque centrifugation results in a depletion of T cells . Moreover, a rather impure lymphoid cell suspension is obtained . In double incubation experiments, T cells (as defined by T cell antigen(s) or rosette formation) and B cells (Fab-bearing cells) were entirely different subpopulations . Allotypes of the a locus could not be detected on the surface of T cells . The results are discussed with respect to genetic coding of antigen receptors on B and T cells. J Endocrinol, 1979 Dec, 83(3), 303 - 10 Varying response to luteinizing hormone of two luteal cell types isolated from bovine corpus luteum; Ursely J et al.; Luteal cell suspensions obtained by enzymatic digestion of pregnant cow corpus luteum were found to be heterogenous and mainly made up of two types of cells of different sizes . The large cells (37 micrometers, average diameter) could be separated from the small ones (18 micrometers, average diameter) by sedimentation at unit gravity in a gradient of Ficoll-bovine serum albumin . A comparative in-vitro study of the synthesis of progesterone by the two types of cells indicated striking differences between them . The average content and the synthesis of progesterone in the absence and presence of a saturating dose of bovine LH after incubation for 2 h were 0.07, 0.12 and 6.9 pg/cell for the small cells and 0.65, 2 and 10 pg/cell for the large ones . Moreover, the sensitivity to low concentrations of LH was 100 to 1000 times higher for the small cells than for the large ones . Oestradiol-17 beta at concentrations ranging from 5 X 10(-10) to 5 X 10(-4) mol/l exerted a dose-dependent inhibition on the stimulation of LH in both cell types . These results suggest a possible involvement of both cell types in the synthesis of progesterone in vivo with a greater contribution by the small cells to stimulation induced by LH . Moreover, it appears that small cell suspensions could be a useful model system for in-vitro studies of the control of the synthesis of progesterone in cow corpus luteum. In Vitro, 1979 Dec, 15(12), 949 - 56 Tapping culture--an improved method for cell suspension culture; Katsuta H et al.; A new culture vessel was designed for cell suspension culture . A silicone-covered magnet bar fixed by one end to the side wall of the bottle was held horizontally a short distance from the bottom . A standard type magnetic stirrer was used . In contrast to the conventional horizontal movement of "stirring" in cultures the bar moves vertically with a "tapping" motion . This improvement resulted in less cell injury, higher rate of cell proliferation and formation of fewer bubbles than in the conventional type . Nine cell types were simultaneously cultivated in tapping, stirring and stationary culture . All cell types proliferated more luxuriously in tapping cultures then in stirring cultures . Serial cultivation of cells in tapping cultures was also successful. J Cell Sci, 1979 Dec, 40, 271 - 9 The effects of EGTA and trypsin on the serum requirements for cell attachment to collagens; Schor SL; Cells growing on plastic or glass surfaces in vitro may be brought into suspension by proteases (e.g . trypsin) or chelating agents (e.g . EGTA) . Trypsin and EGTA remove different quantities and types of molecules from cell surfaces . Previous studies have revealed that when confluent cultures of either BHK or PyBHK cells are brought into suspension by exposure to trypsin, foetal calf serum (or fibronectin) is required for cell attachment to films of denature type I collagen, but not to 3-dimensional gels of native collagen fibres . In this communication the serum requirements for the attachment of BHK and PyBHK cells to collagen substrata have been examined as a function of (a) the method used to prepare the cell suspension (EGTA or trypsin), and (b) cell density . Data are presented consistent with the view that cell surface-associated fibronectin is able to mediate cell attachment directly to films of denatured collagen. Biull Eksp Biol Med, 1979 Dec, 88(12), 693 - 5 {Suppression of delayed hypersensitivity in mice receiving a massive dose of xenogeneic erythrocytes}; Chernousov AD et al.; The injection of 6x109 sheep red blood cells to mice suppresses delayed-type hypersensitivity (DTH) in situ and activates spleen cells which prevent sensitization of recipients . Preliminary thymectomy of donors and the treatment of cell suspensions with anti-T-globulin abolish suppression of DTH . Pretreatment of mice with low doses of cyclophosphamide (CY) enhances antibody formation and DTH . Higher doses of CY increase the DTH reaction but inhibit antibody formation . The data obtained allow to conclude that suppression of DTH is due to the activity of short-living, intensively proliferating cells of thymic origin and possibly to B cells. Appl Environ Microbiol, 1979 Dec, 38(6), 1147 - 52 Sublethal stress in Escherichia coli: a function of salinity; Anderson IC et al.; Sublethal stress in Escherichia coli was detected in various test media after exposure (in vitro) to seawater of various salinites . Stress was measured with an electrochemical detection technique and a beta-galactosidase assay . Test media included EC medium, medium A-1, and tryptic soy broth modified to contain lactose for beta-galactosidase assay experiments . Stress was defined as the difference between a predicted electrochemical response time calculated for unstarved cells from a standard curve and the observed electrochemical response time for cells starved in seawater . The higher the salinity, the greater the stress for all test media examined . Stress was most pronounced in EC and was attributed primarily to initial die-off of starved cells exposed to the test medium at the elevated temperature of 44.5 degrees C . Lag time and growth rates in test media were not significantly affected by salinity . beta-Galactosidase specific activity, assayed in starved cells after transfer to an induction medium at 44.5 degrees C for 150 min, was inversely related to the salinity of the starved cell suspension . The consequences of these observations with respect to coliform enumeration methods are discussed. Arch Int Physiol Biochim, 1979 Dec, 87(5), 925 - 34 Cytochalasin E-induced oxidative metabolism in polymorphonuclear leukocytes; Heyneman RA; The extent of the cyanide-resistent oxidative burst of polymorpho-nuclear leukocytes after stimulation with cytochalasin E was shown to depend markedly on the osmolarity of the cell-suspension medium . With granulocyte concentrations up to 2 X 10(6) cells/ml, optimal oxygen consumption and releases of H2O2 and superoxide anions were reached at 180 mOsmol and 2 X 10(-5) M cytochalasin E . After removal of unbound activator, the cellular oxidative activity remained unaltered and continued to depend on the used osmotic conditions . It is proposed that binding of cytochalasin to the plasma membrane induces an irreversible activation of the oxidative system, whereas the resulting metabolic activity depends on conformational changes in the plasma membrane. J Membr Biol, 1979 Nov 30, 50(3-4), 311 - 29 Manganese as a calcium probe: electron paramagnetic resonance and nuclear magnetic resonance spectroscopy of intact cells; Getz D et al.; When Lettree cells are exposed to Mn2+, the cation becomes associated with cells in two ways: in a relatively loose and mobile manner that gives a six-line EPR spectrum designated Mnb*, and in an immobile, relatively tight manner that gives no detectable EPR specrtum, designated Mnb . Mnb* is probably on the surface of cells; most Mnb is probably inside cells . NMR measurements of Lettree cell suspensions show two water proton relaxation rates and confirm the existence of cell-associated Mn . Human erythrocytes, on the other hand, bind no Mn2+ under these conditions, as judged by EPR and NMR measurements . Virally-treated Lettree cells show an increase in Mnb (but not in Mnb*) . They also show a third water proton relaxation rate. Prikl Biokhim Mikrobiol, 1979 Nov-Dec, 15(6), 883 - 8 {Determination of cytochromes in chloroplasts of Chlamydomonas reinhardii mutants}; Roshchina VV et al.; A method for qualitative determination of cytochromes of the wild-type and mutant strains of Chalmydomonas reinhardii was developed . The effect of different techniques of cell disruption (ultrasound, Triton X-100, acetone, etc) on detection of cytochrome maxima in the oxidized minus reduced difference spectra was studied on a comparative basis . The development of the stable difference spectrum of the disrupted cell suspension was shown to be a function of incubation time in the presence of an oxidizing or a reducing agent . Cytochromes of the wild-type and 10 nonphotosynthesizing mutants were determined . Four mutants with lesions in cytochromes B559 and C553 were detected . Mutants with lesions in the reaction center of photosystem 2 were found to have a substantially reduced content of cytochrome B559, whereas those with normal photosystems--of cytochrome C553. Microsc Acta, 1979 Nov, 82(3), 251 - 3 Dissociation of rat colonic mucosa into single epithelial cells and their microscopic visualization; Herp A et al.; Repeated incubation of rat colonic mucosa with phosphate-buffered mannitol yields morphologically intact single cell suspensions which are easily visualized microscopically with trypan blue . Cell dimensions are determined by means of a Nikon microcomparator. J Clin Pathol, 1979 Nov, 32(11), 1180 - 3 A sensitive single reverse passive haemagglutination test for detecting both HBsAg and anti-HBs; Barbara JA et al.; A trial of a modified reverse passive haemagglutination test for HBsAg using a 0.1% cell suspension instead of the recommended 1% showed an approximately eight-fold increase in detection sensitivity . The test can be performed within 30 minutes and lends itself to mass screening techniques . Confirmation tests can be done using the 0.1% method . In addition, the same serological plates and cells used for HBsAg screening can then be used to screen for high-titre anti-HBs . This makes the overall screening for both HBsAg and high-titre anti-HBs donors cheap and convenient. Eur J Biochem, 1979 Nov 1, 101(1), 225 - 33 Purification and properties of strictosidine synthase, the key enzyme in indole alkaloid formation; Treimer JF et al.; A new enzyme, strictosidine synthase, which catalyzes the synthesis of 3-alpha(S)-strictosidine from tryptamine and secologanin was isolated from the soluble protein extract of Catharanthus roseus cell suspension cultures and was purified approximately 50-fold by ammonium sulfate fractionation, column chromatography on DEAE-cellulose . Ultrogel AcA34 and isoelectric focusing . The apparent molecular weight of the enzyme was 34000 . The pH optimum was 6.8, apparent Km values for tryptamine and secologanin were 2.3 mM and 3.4 mM respectively for the enzyme to synthesize strictosidine . Strictosidine synthase shows high substrate specificity . No apparent cofactor requirement could be demonstrated . Of several enzyme inhibitors tested, only p-chloromercuribenzoate inhibited the enzyme . The enzyme was relatively stable and could be stored at -20 degrees C for periods of up to 1 year without appreciable loss of catalytic activity . The enzyme was demonstrated to occur in suspension cultures of 15 different species belonging to 9 different genera of the indole-alkaloid-producing subfamily Plumerioideae of the Apocynaceae family . This enzyme is responsible for the synthesis of strictosidine the key intermediate in the formation of the majority of monoterpenoid indole alkaloids occurring in the plant kingdom. Blut, 1979 Nov, 39(5), 333 - 44 Red cell filtration by paper filters; Staubli M et al.; A method for the assessment of red cell filterability with paper filters is described . Two milliliters of a washed red cell suspension with a hematocrit of 50% was filtered through a filter paper cone in a glass funnel and the filtration half-time (FT1/2) was measured . The filter papers were calibrated by previous measurement of the filtration time for the suspending solution, the calibration time (CT) . A linear correlation between CT and FT1/2 was found for individual red cell suspensions (p less than 0.001) as well as for a normal population (p less than 0.001) . Consequently, the index FT1/2/CT should represent a more reproducible parameter than FT1/2 and experimentally this appears to be the case . In fact, elimination of erratic values in a normal population was realized . Furthermore, discrimination between normal and abnormal values was improved and the coefficient of variation of single measurements passed from 28.4 to 9.4% . The reproducibility and reliability of this extremely simple method are therefore decisively enhanced without complication of the procedure. Int J Radiat Biol Relat Stud Phys Chem Med, 1979 Nov, 36(5), 435 - 51 Effect of salt solutions on the radiosensitivity of mammalian cells as a function of the state of adhesion and the water structure; Moggach PG et al.; The radiation isodose survival curve of attached Chinese hamster (V79) cells, subjected to a wide concentration range of salt or sucrose solutions, is characterized by two maxima separated by a minimum . Cells are radioprotected at the maxima (high and low hypertonic salt concentrations) while they are radiosensitized at the minimum (intermediate hypertonic salt concentrations) . Both cations and anions can alter the cellular radiosensitivity above and beyond the (osmotic) effect observed for cells treated with sucrose solutions . However, the basic curve shape, except in the case of sulphate salts, remains the same . When these experiments are repeated with single cells in suspension, the isodose survival curve is quite different in that high salt concentrations (greater than 0.9 M) do not protect cells in suspension unlike the case with attached cells . The curve shape is also altered in that the second maximum is absent with many salt solutions . If multicellular spheroids are used for these experiments, the data resemble those for single cell suspensions rather than for attached cells . The radiation survival data for cells in suspension in salt solutions correlate with water proton spin-lattice relaxation time (T1) and, in hypo- and iso-tonic solutions, with cell volume. Anal Quant Cytol, 1979 Nov-Dec, 1(3), 228 - 32 The use of percent volume analysis in Coulter sizing of cells in gynecologic cytopathologic cell suspensions; Leary JF et al.; The Coulter Counter model TA II was evaluated as a gynecologic cytodiagnostic tool . The instrument produces particle size histograms as plots of percent volume versus particle volume in 16 channels with each channel corresponding to twice the volume of the next lower channel . Cell volume distributions were obtained using cell suspensions from patients with cervicitis, inflammation, dysplasia or carcinoma and were compared with those from normal subjects . Although this method is superior to other methods of volume distribution analysis, it does not satisfy the stringent requirements of a cervical cancer mass screening method. Tsitologiia, 1979 Nov, 21(11), 1372 - 3 {Method of cell cultivation in microdishes}; Kovalev SP; A method is proposed of cell cultivation on the microscopic glass . Glass rings with inner diameter of 6 mm are fixed on prepared glasses . A simple of 0.2 ml cell suspension is poured into so-formed microcups . The cultivation is carried out in hermetic Petri dishes under conditions of the 5% CO2 atmospheric saturation at 37 degrees C. Br J Cancer, 1979 Nov, 40(5), 689 - 700 Association of host immunoglobulins with solid tumours in vivo; James K et al.; Using a direct radioimmune antiglobulin technique and a competitive double-antibody radioimmune assay, we have demonstrated the presence of appreciable amounts of host immunoglobulins on the surface and in extracts of cell suspensions from freshly excised solid tumours . IgA appeared to have the greatest concentrations from freshly excised solid tumours . IgA appeared to have the greatest concentration, followed in turn by IgM congruent to IgG2a greater than IgG1 congruent to IgG2b greater than IgG3 . The amount of immunoglobulin appeared to be influenced by the tumour under investigation and its mode of maintenance . It could also be increased by the administration of C . parvum but was not significantly influenced by the T-cell status of the host. Endocrinology, 1979 Nov, 105(5), 1183 - 90 Sulfhydryl reagent-induced insulin release and 45Ca++ fluxes in Syrian hamster insulinoma cells; Erlichman J et al.; Dispersed single cell suspensions of Syrian hamster insulinoma cells were used to study the effects of a variety of sulfhydryl-binding reagents on insulin release and 45Ca++ flux . Incubation of cells with several organic mercurials resulted in a rapid increase in 45Ca++ uptake as well as increased efflux in cells which had been prelabeled with 45Ca++ . Concomitant with increased calcium uptake was a 4- to 5-fold increase in insulin released into the medium . Incubation with alkylating reagents such as iodoacetamide and N-ethyl maleimide or dithiol reagents such as 5,5'-dithiobis (2-nitrobenzoic acid) failed to stimulate either 45Ca++ flux or insulin release . Elimination of medium calscium or preincubation of cells with N-ethyl maleimide resulted in approximately 50% inhibition of mercurial-induced insulin release from these cells . 8-(N,N2-diethylamino)Octyl-3,4,5-trimethoxybenzoate or alpha-isopropyl-alpha {(N-methyl-N-homoveratryl)-gamma-aminopropyl}3,4,5'-trimethoxyphenylacetonitrite hydrochloride, agents which block potassium (40 mM)-stimulated calcium flux and insulin release, failed to inhibit mercurial-induced calcium flux or insulin secretion . These results indicate that sulfhydryl-binding reagents, through their interaction with critical thiol groups, promote insulin release in these insulinoma cells by inducing changes in calcium fluxes . It is possible that these thiol groups regulate calcium metabolism and, thus, insulin release under physiological conditions. J Cell Biol, 1979 Nov, 83(2 Pt 1), 271 - 83 Characterization of gastric mucosal membranes . X . Immunological studies of gastric (H+ + K+)-ATPase; Saccomani G et al.; Gastric mucosal homogenates from hog were fractionated by differential and density gradient centrifugation and free-flow electrophoresis . The two major membrane fractions (FI and FII) thus obtained are distinct both enzymically and in terms of transport reactivity . This heterogenicity extends to their antigenic activity . Purified antibodies which were raised against the K+-ATPase-containing H+ transport fraction FI were of two types: inhibitory and non-inhibitory . Inhibitory antibodies reduced the K+-ATPase activity by approximately 80% and the K+-p-nitro-phenylphosphatase activity by approximately 40% in a concentration-dependent manner, while the small Mg++-dependent component of the enzyme activity was unaffected . Antibodies inhibiting the K+-ATPase also inhibited H+ transport . These antibodies did not cross-react with the other major membrane fraction isolated by free-flow electrophoresis, FII, and gave a single band on rocket immunoelectrophoresis . Antibodies against this FII fraction also did not react with the K+-ATPase and were heterogeneous, giving at least four bands with rocket immunoelectrophoresis and inhibiting both the 5'-nucleotidase and Mg++-ATPase of this fraction . Immunofluorescent staining of tissue sections showed that the FI was derived from the parietal cell of gastric tissue and was localized to the supranuclear area of the cell . Staining of isolated rat gastric cell suspensions by FI antibodies confirmed the selectivity of the antibody and showed a polar, plasma membrane localization . FII antibodies also largely stained the parietal cells in tissue sections . In the 16 hog tissues tested, FI antibodies cross-reacted only with gastric fundus, thyroid and weakly with thymus . Immunoelectronmicroscopy showed that FI antibodies reacted strongly with the secretory membrane at the apical cell surface of the parietal cells and at the secretory canaliculi, weakly with the apical surface of the zymogen cell, and not with the basal-lateral surface of the cells . Thus, the protontranslocating ATPase is localized in the parietal cells and in the region postulated to be the site of acid secretion. Cancer, 1979 Nov, 44(5), 1622 - 8 Studies of mixed lymphocyte reactions, surface B cell antigens, and intracytoplasmic immunoglobulins in "null cell" acute lymphocytic leukemia; Davey FR et al.; Lymphoblasts from "null cell" acute lymphocytic leukemia (ALL) were analyzed for the pattern of proliferation displayed in a mixed lymphocyte reaction (MLR), the presence of a B cell surface antigen, and for the presence of intracytoplasmic immunoglobulin (ICIg) . "Null cell" ALL was defined by cytologic and cytochemical criteria and by the absence of spontaneous rosette formation and surface membrane immunoglobulin in cell suspensions of the malignant lymphocytes . In eleven of fourteen patients the proliferative characteristics of lymphoblasts in the MLR were similar to those observed with normal B enriched lymphocytes . In eleven cases studied, anti-B cell serum reacted with a majority of the lymphoblasts . None of the ten cases examined displayed ICIg in the lymphoblasts . We conclude that the "null" lymphoblast from most cases of ALL is a B cell in an early stage of development. Infect Immun, 1979 Nov, 26(2), 448 - 52 Ability of enriched immune T cells to confer resistance in hamsters to infection with Treponema pertenue; Chan JK et al.; This investigation presents the first direct evidence that T cells are involved in resistance to challenge with Treponema pertenue . Enriched T cells from immune hamsters were obtained by sequential filtration through glass and nylon-wool columns . This procedure removed the majority of functional antibody-producing and immunoglobulin-bearing cells . The fractionated cell suspensions were less responsive to stimulation by phytohemagglutinin, lipopolysaccharide, and dextran sulfate, but they were enriched with antithymocyte-sensitive cells and were more responsive to stimulation with concanavalin A . Hamsters receiving fractionated or unfractionated immune cells had no cutaneous lesions 21 days after infection and had significantly lower lymph node weights and fewer treponemes per node than hamsters that received fractionated or unfractionated normal cells . Resistance was transferred with immune cell suspension enriched in T cells despite an absence of anamnestic antibody response to specific treponemal antigens. J Biochem (Tokyo), 1979 Nov, 86(5), 1291 - 300 Preparation and characterization of an active lysozyme derivative: Kyn 62-lysozyme; Yamasaki N et al.; A novel method for the preparation of Kyn 62-lysozyme, in which tryptophan 62 is replaced by kynurenine, is reported . Hen egg-white lysozyme was ozonized in aqueous solution to yield one N'-formylkynurenine residue and deformylated with hydrochloric acid in frozen solution at -10 degrees C . Crude Kyn 62-lysozyme was purified by affinity and Bio Rex 70 chromatography successively . Kyn 62-lysozyme retains affinity for chitin and is essentially an active enzyme with a slightly weakened but distinct catalytic activity . After this modification, the enzyme activity was changed differently depending on the kind of substrate . At the individual optimum pH's, lytic activity was largely retained (80% active), but the catalytic efficiency for hydrolyzing glycol chitin was relatively low (30% active) . Lysis of M . lysodeikticus cell suspensions was optimally catalyzed by Kyn 62-lysozyme at pH 6.2 and at 0.088 ionic strength . These values are lower by 1.3 pH unit and 0.04 ionic strength, respectively, than those of intact lysozyme . The optimum pH and ionic strength for the hydrolysis of neutral substrates were scarcely affected . These results suggest the significance of electrostatic interaction in the lysis of lysozyme . Relatively limited loss of activity induced by modification of the 62nd residue, which is thought to participate directly in the binding of the substrate at subsite C, is discussed on the basis of the similarity of side chain structure in tryptophan and kynurenine. Int J Cancer, 1979 Oct 15, 24(4), 450 - 4 Contribution of macrophages to interactions between tumor cells and other tissues assessed in an in vitro system; Maslow DE et al.; The modification of the tissues surrounding solid tumors is usually attributed exclusively to interactions between the normal tissues and the cancer cells in the tumor, in spite of the fact that tumors contain many different types of non-cancer cells including macrophages . As an experimental model for some facets of the cellular interactions between tumor and normal tissues, we have assessed the individual contributions of macrophages and cancer cells to the differential inhibition of neural retina (NR) cell aggregation by co-culturing NR cells with small numbers of macrophages or cancer cells alone, as well as with both natural (ascites tumors) and artificial mixtures . Cells from human colonic adenocarcinoma inhibited NR aggregation to a greater extent than normal colon mucosa . Aggregation of NR cells was inhibited by macrophages from mice and rats, and to a greater extent by cancer cell suspensions of mouse Ehrlich and rat Walker 256 lines from spinner culture or in the ascites form . Combinations of macrophages and cancer cells indicated that their inhibitory effects were neither additive nor synergistic . Cell-free media from macrophages and cancer cell cultures were equally effective inhibitors of aggregation . The results suggest that the interaction between a malignant tumor and the surrounding normal tissue can be modified by cancer cells, tumor macrophages and their products. Nature, 1979 Oct 11, 281(5731), 497 - 9 Evidence for a charge-shift electrochromic mechanism in a probe of membrane potential; Loew LM et al.; Extrinsic optical probes have become important tools for monitoring membrane potential, with probes now available for many tissue or cell suspension systems . In each case that has been studied in detail, it seems that the mechanism involves a shift in the equilibrium population of the probe from one chemical environment to another in response to the transmembrane potential; the environments perturb the probe's spectrum differently . As this indirect mechanism involves a redistribution of dye between chemical environments that are likely to vary if a given probe is transferred from one membrane to another, a potential probe that is effective and calibrated for all membrane systems has not been realised . We present here evidence for a direct response of a probe chromophore to the electric field across membrane systems . The results suggest it might be possible to develop a universal set of membrane probes. J Membr Biol, 1979 Oct 5, 50(1), 23 - 41 HCO3-/Cl- exchange across the human erythrocyte membrane: effects of pH and temperature; Obaid AL et al.; Changes in extracellular pH (PH0) in red cell suspensions were monitored in a stopped-flow rapid reaction apparatus under conditions where dpH0/dt was determined by the rate of HCO3-/Cl- exchange across the membrane . Experiments were performed at 5 degrees C less than T less than 40 degrees C using either untreated cells or cells exposed to 0.11 mM SITS (4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid) . Although SITS exposure reduced the rate of exchange by 90%, both untreated and SITS-treated cells are similarly affected by changes in pH0 and temperature . The rate of HCO3-/Cl- exchange exhibits a minimum at about pH0 5 and a maximum at about pH0 7.4 at all temperatures . A transition temperature of 17 degrees C was observed in the Arrhenius relationship for all pH0 . The activation energies (Ea) in kcal/mol are 19.6 below and 11.7 above 17 degrees C for 5 less than pH0 less than 8 . These findings, similar to those reported for Cl- self-exchange, suggest that: (i) a change in the rate-limiting step for HCO3-/Cl- exchange occurs at 17 degrees C, possibly due to an altered interaction between the transport pathway and membrane lipids; (ii) the carrier system can be titrated by either H+ or SITS from the outside of the membrane, but the untitrated sites continue to transport normally; (iii) the pH0 dependence of the rate of exchange is consistent with the titratable carrier having its most alkaline pK in the range expected for amino groups; and (iv) below pH0 5, the nature of the exchange is markedly altered. Br J Exp Pathol, 1979 Oct, 60(5), 499 - 506 Comparative studies of the metastatic potential of three transplantable rat mammary carcinomas of spontaneous origin; Willmott N et al.; The metastatic potential of 3 spontaneously arising mammary carcinomas (Sp4, Sp15 and Sp22) has been examined when transplanted in the form of a viable cell suspension into the tissue of origin . Primary tumours were excised at different times after implantation and it was found that the metastatic potential of the immunogenic tumour Sp4 was directly correlated with the size of the primary tumour when excised . By contrast, the incidence of metastases from the non-immunogenic tumours Sp15 and Sp22 was similar irrespective of the size of the primary tumour on excision . The pattern of metastasis also differed between the tumours, although here there was no relation to immunogenicity . Thus, resection en bloc of large primary Sp4 or Sp15 tumours plus regional lymph nodes could be completely curative, signifying initial spread of tumours via the lymphatics and only subsequently via the blood stream . On the other hand, resection en bloc of primary Sp22 tumours plus regional lymph nodes at a similar stage of primary tumour development was never curative, signifying early spread via the blood stream . Other studies showed that the metastatic potential of mammary carcinoma Sp4 was an innate characteristic of the tumour and not related to the tissue of implantation since in addition to metastasizing from the mammary pad it metastasized when implanted either s.c . or intradermally in a region devoid of mammary tissue . Furthermore, a rat sarcoma Mc7 showed a negligible tendency to metastasize when implanted either in the mammary pad or in the s.c . tissue, where it had been induced with methylcholanthrene. J Immunol, 1979 Oct, 123(4), 1778 - 80 Rapid magnetic purification of rosette-forming lymphocytes; Owen CS et al.; High gradient magnetic separation, which as previously been shown effective in extracting erythrocytes from a flowing cell suspension, has been used to separate rosetted and unrosetted human peripheral blood lymphocytes . The hemoglobin in the sheep red cells used to form rosettes was first oxidizied to the paramagnetic methemoglobin form . Samples of 50 x 10(6) lymphocytes could be processed in 10 min under sterile conditions with greater than 90% purity of the rosetted cell fraction and maintenance of T cell function in mixed lymphocyte cultures. Am J Pathol, 1979 Oct, 97(1), 17 - 41 alpha-Naphthyl acetate esterase activity--a cytochemical marker for T lymphocytees . Correlation with immunologic studies of normal tissues, lymphocytic leukemias, non-Hodgkin's lymphomas, Hodgkin's disease, and other lymphoproliferative disorders; Pinkus GS et al.; Cytochemical identification of T lymphocytes on the basis of alpha-naphthyl acetate esterase (NAE) activity was compared with immunologic markers for cell suspensions and/or cryostat sections of 113 specimens . Nonneoplastic tissues (peripheral blood, lymph nodes, spleens, tonsils, thymus, and pleural fluid) and specimens from various lymphoproliferative disorders, including acute and chronic lymphocytic leukemia, lymphosarcoma cell leukemia, hairy cell leukemia, non-Hodgkin's lymphomas of B-and T-cell types, and Hodgkin's disease, were evaluated . T (E-rosetting) cells demonstrated several patterns of NAE reactivity: 1) a strong globular reaction product, the most specific pattern for T-cell identification, 2) granular cytoplasmic staining, or 3) no reactivity . B lymphocytes revealed a granular pattern of NAE staining, were devoid of enzyme, or, in rare instances, exhibited strong NAE activity . Percentages of lymphoid cells with strong (globular) NAE activity closely paralleled T-cell (E-rosette) values in the majority of cases, with the best correlations observed for peripheral blood studies . However, discordant results were noted for some neoplastic and nonneoplastic tissues, including cases of T-cell lymphoma or leukemia . Markedly discrepant results were noted for thymic lymphocytes, most of which revealed E-rosette formation and weak or absent NAE activity . Lymph nodes involved by Hodgkin's disease demonstrated a heterogeneous pattern of staining in E-rosetting cells and in Reed-Sternberg variants . Cryostat section studies of reactive lymph nodes and nodular lymphomas demonstrated strong NAE staining in lymphoid cells of T-cell (interfollicular, internodular) areas, with little or no positivity in follicles or nodules (B-cell areas) . NAE staining patterns further suggested that T cells comprise part of the follicular cuff and possibly represent a minor population of some neoplastic nodules . Although NAE determinations do not represent a consistently reliable alternative to immunologic methods for T-cell identification, this easily applicable cytochemical marker is complementary to other techniques in assessing neoplastic or nonneoplastic tissues, particularly cryostat sections . (Am J Pathol 97:17--42, 1979). J Immunol, 1979 Oct, 123(4), 1818 - 21 Presence of T cell-associated surface antigens on murine NK cells; Pollack SB et al.; The expression of T cell-associated surface antigens on natural killer (NK) spleen cells of C57BL/6 mice was evaluated by cytotoxic depletion experiments with alloantisera prepared against the Thy 1, Ly 1, Ly 2, Ly 5, Ly 6, and NK 1 antigens . The NK activity of these nonimmunized spleen cells for YAC-1 leukemia cells was dramatically reduced by antisera to the Ly 5 and NK 1 antigens . Variable results were obtained with anti-Ly 6 sera--certain pools of this antiserum decreased the NK activity, whereas other pools showed only negligible effects . The NK activity of the same cell suspensions was not affected by antisera to the Thy 1, Ly 1, and Ly 2 antigens . In parallel tests the T cell-associated cell surface antigens of alloimmune T killer cells were similarly evaluated by cytotoxic depletion experiments . In this case, the activity of these cells was consistently diminished by antisera to the Thy 1, Ly 2, Ly 5, and Ly 6 antigens, but not by antisera to the Ly 1 and NK 1 antigens . On this basis it was concluded that the NK cells expressed a restricted subset of T cell-associated alloantigens and therefore may have been derived from the T cell lineage of lymphocytes. J Clin Microbiol, 1979 Oct, 10(4), 533 - 7 Comparison of rates of virus isolation from leukocyte populations separated from blood by conventional and Ficoll-Paque/Macrodex methods; Howell CL et al.; One hundred fifty-two blood specimens, largely from immunocompromised patients, were collected in heparinized Vacutainer tubes and divided into paired aliquots of equal volume . Buffy-coat preparations, containing mixed leukocyte and separate mononuclear and polymorphonuclear leukocyte populations were obtained by treatment of blood with conventional and Ficoll-Paque/Macrodex (F-P/M) methods . The development of cytopathic effect in monolayers of WI-38 fibroblasts inoculated with cell suspensions derived from the two methods was used to assess virus infectivity . Twice as many virus isolations were obtained using F-P/M . Of those viruses isolated by both conventional and F-P/M, the development of cytopathic effect was more extensive using the latter method . Moreover, a greater variety of viruses was isolated using F-P/M method, as compared to the conventional method . The F-P/M method is no more time consuming than conventional procedures, is readily adaptable for use in the diagnostic virology laboratory, requires only minimal additional cost, and is a particularly suitable and effective means of monitoring viremia. J Biochem (Tokyo), 1979 Oct, 86(4), 971 - 7 Probable sites of action of cyclic adenosine 3',5'-monophosphate in the induction of phosphodiesterase in Dictyostelium discoideum; Yamasaki F et al.; The sites of action cyclic adenosine 3',5'-monophosphate (cAMP) in phosphodiesterase {EC 3.1.4.17} induction in Dictyostelium discoideum were studied . When cAMP was added to the cell suspension from the start of the incubation, the effect of the cyclic nucleotide on the cellular enzyme-induction did not appear for 30 min, then occurred abruptly . From experiments on the addition of cAMP to the cell incubation mixture at various times, preparations for the synthesis of the enzyme appear to occur during the period from 20 min to 30 min after the start of the incubation . After the addition of cycloheximide at 30 min, the enzyme was degraded very rapidly . The half-life of phosphodiesterase was roughly 21 min in the presence of cAMP, and 12 min in its absence . The degradation rates became approximately the same on removing cAMP . When daunomycin and actinomycin D were added to cells previously stimulated with cAMP, phosphodiesterase was still synthesized at a higher rate than in cells not pretreated with cAMP . These results suggest that cAMP acts at two sites at least, i.e., on enzyme synthesis at the transcription level, and in suppressing the degradation of phosphodiesterase. J Anat, 1979 Oct, 129(3), 541 - 59 Dissociation of adult mammalian heart into single cell suspension: an ultrastructural study; Nag AC et al.; Adult rat heart was dissociated into a single cell suspension by a perfusion technique which used 0.05% collagenase and 0.1% hyaluronidase in Krebs-Ringer phosphate buffer (KRP) . The non-muscle cells of the suspension were separated from the myocytes by centrifugation through 3% Ficoll solution in KRP with 0.01 mM Ca2+ . An approximately 90% pure suspension of isolated single muscle cells was obtained with this method . The effects of the successive steps in the dissociation procedure on the ultrastructure of the heart were studied by scanning and transmission electron microscopy . After 30 minutes of enzyme digestion, dissociation of the inner endothelial lining of the ventricle into single cells or small groups of cells became apparent . In addition, the underlying cardiac skeleton began to disintegrate and linear arrays of cardiac muscle cells were observed . After 45 minutes of enzyme digestion the number of released single cells was higher because of the separation of intercalated discs . The majority of non-muscle cells were by now dissociated from the surfaces of muscle cells . Widening of the lateral intercellular spaces between the myocardial cells was associated with separation of desmosomes . In some regions of the heart, intact desmosomes, fasciae adherentes and gap junctions were observed even though lateral intercellular spaces had widened greatly . The majority of myocardial cells had become separated from one another after 60 minutes of enzyme digestion . Separation of gap junctional sites took place in two ways: (1) by 'unzipping' them through enzyme action; (2) by tearing them mechanically . Gap junction remnants were sometimes observed in a vesiculated state within the cell . The dissociation of the heart was ineffective when perfused with media containing 1.0 or 2 mM Ca2+ . Alcian blue treatment after 60 minutes of enzyme digestion revealed that the basement membrane, and its accompanying collagen fibrils, was still present on the plasma membrane of dissociated single cells . The isolated myocardial cells retained their normal morphological characteristics . This study has enabled us to understand in detail how dismantlement of highly ordered adult cardiac tissue into a single cell suspension takes place . Cell suspensions of this type should be invaluable in the study of metabolic and synthetic activities in adult myocardial cells. Biochim Biophys Acta, 1979 Sep 21, 556(2), 278 - 91 Biochemical and ultrastructural study of the disruption of blood platelets by streptolysin O; Launay JM et al.; The membrane-damaging protein toxin, streptolysin O, proved highly lytic on human, guinea-pig and rabbit platelets . About 15 molecules of toxin were sufficient to lyse one cell . Platelet disruption was assessed by electron microscopy, clearing of cell suspensions and assay of lactate dehydrogenase, serotonin, monoamine oxidase and glutathione peroxidase released in the extracellular fluid . This egress reflected the damage of both plasmic and organelle membranes . A quantitative study of lactate dehydrogenase and serotonin liberation taken as respective markers of the cytosol and dense bodies was undertaken as a function of toxin concentration . No platelet aggregation or shape change was elicited by streptolysin O . The ghosts resulting from platelet lysis retained properties of the native membrane such as aggregability and serotonin uptake . Dense bodies were easily separated after gentle disruption of the plasmic membrane by small amounts of toxin . Platelet lysis by streptolysin O proved a useful procedure for the determination of protein content, enzyme activities and serotonin assay on the same lysate in contrast to usual methods. World J Surg, 1979 Sep 20, 3(5), 641 - 50 Heterotransplantation of human gastric carcinomas into nude mice; Nakatani K et al.; A total of 33 specimens of human gastric carcinoma were used for transplantation into nude mice . Initital tumor "take" was accomplished in 15 of the 33 tumors, and the transplantability rate was 45.5% . Transplantability correlated with histologic type, but not with clinical stage or Borrmann's classification . The transplantability rate of differentiated carcinomas, such as well-differentiated tubular adenocarcinoma, moderately differentiated tubular adenocarcinoma, and papillary adenocarcinoma was greater than that of poorly differentiated tumors, such as poorly differentiated adenocarcinoma and mucinous adenocarcinoma . The growth patterns of transplanted tumors were divided into 3 types: rapid, slow, and persistent . There were no specific relationships between growth pattern and histologic type . All histologic types, except signet ring cell carcinoma, could be transplanted serially . Tumor growth became rapid after serial transfer . However, the original histology of these tumors was unchanged . No invasion or metastases were encountered . Intraperitoneal injection of a tumor cell suspension, prepared from subcutaneous transplants of a poorly differentiated adenocarcinoma of Borrmann type III, grew in an ascites form, with invasion and metastasis . Ascitic fluid accumulated within 3--6 weeks after injection . Subsequently, intravenous injection of ascites fluid produced metastases in nude mice . The histology of the subcutaneous tumor was similar to that of the original tumor from the patient. Biochem J, 1979 Sep 15, 182(3), 697 - 705 Mechanism of the stimulation of serine and alanine transport into isolated rat liver cells by bicarbonate ions; McGivan JD; 1 . Bicarbonate ions stimulate the transport of serine and alanine into isolated hepatocytes . 2 . The effect of bicarbonate is to increase the Vmax . of the transport process without changing the apparent Km . 3 . The intracellular pH was estimated from the distribution of the weak base methylamine and the weak acid 5,5'-dimethyloxazolidine-2,4-dione (DMO) across the plasma membrane . 4 . The addition of bicarbonate to a cell suspension caused the internal pH to become more acid . 5 . The initial rate of serine, alanine and glycine transport was a linear function of the initial difference in pH across the membrane . 6 . It is concluded that bicarbonate activates the transport of these amino acids primarily by increasing the pH difference across the plasma membrane . 7 . It is suggested that the uptake of serine together with Na+ ions occurs in exchange for H+ ions, which are translocated outwards on the same carrier system . Some preliminary evidence consistent with this model is presented. Biochem J, 1979 Sep 15, 182(3), 687 - 96 Quantitative analysis of proton-linked transport systems . The lactose permease of Escherichia coli; Booth IR et al.; Evidence is presented that lactose uptake into whole cells of Escherichia coli occurs by symport with a single proton over the range of external pH 6.5--7.7 . The proton/lactose stoicheiometry has been measured directly over this pH range by comparison of the initial rates of proton and lactose uptake into anaerobic resting cell suspensions of E . coli ML308 . Further, the relationship between the protonmotive force and lactose accumulation has been studied in E . coli ML308-225 over the range of external pH 5.9--8.7 . At no point was the accumulation of the beta-galactoside in thermodynamic equilibrium with the protonmotive force . It is concluded that the concentration of lactose within the cell is governed by kinetic factors rather than pH-dependent changes in the proton/substrate stoicheiometry . The relevance of these findings to the model of pH-dependent proton/substrate stoicheiometries derived from studies with E . coli membrane vesicles is discussed. Biochim Biophys Acta, 1979 Sep 3, 586(3), 453 - 63 Stimulation of de novo synthesis of L-phenylalanine ammonia-lyase in relation to phytoalexin accumulation in Colletotrichum lindemuthianum elicitor-treated cell suspension cultures of french bean (Phaseolus vulgaris); Dixon RA et al.; (1) The regulation of the accumulation of the isoflavonoid-derived phytoalexin phaseollin in cell suspension cultures of Dwarf French Bean (Phaseolus vulgaris/ has been investigated . (2) An elicitor preparation from cell walls of Colletotrichum lindemuthianum, the causal agent of anthracnose disease of French bean, caused a marked accumulation of phaseollin in the cultures . The elicitor induced phaseollin accumulation to a level of 60% that obtained with the artificial elicitor autoclaved ribonuclease A and was maximally active at a concentration (weight basis) of at least 50 times lower than required for maximal response to ribonuclease . (3) Elicitor preparations from cell walls of Phytophthora megasperma var . sojae, a fungal pathogen of soybean, and Botrytis cinerea, the common grey mould, were much less effective than the C . lindemuthianum wall-released elicitor . (4) There was a marked but transient increase in the extractable activity of phenylalanine ammonia-lyase, the enzyme catalysing the first reaction in the biosynthesis of phaseollin from L-phenylalanine, in response to the elicitor from C . lindemuthianum . (5) Comparative density labelling with 2H from 2H2O indicated that the elicitor stimulates de novo synthesis of phenylalanine ammonie findings provide the basis of a scheme for elicitor induction of phytoalexin accumulation. J Lipid Res, 1979 Sep, 20(7), 914 - 8 Measurement of cholic acid synthesis and secretion by isolated rat hepatocytes; Whiting MJ et al.; Liver cells isolated from normal and cholestyramine-treated rats were incubated as cell suspensions for up to 4 hr in a simple, defined medium . The bile acid concentration in cells plus cell medium was determined by gas-liquid chromatography . Normal hepatocytes synthesized cholic acid at an initial rate of 0.25 nmol/mg cell protein per hr, which is comparable to rates reported from in vivo methods . This rate was increased more than 4-fold when rats were fed a cholestyramine-containing diet for 7 days prior to liver cell isolation . Although cholic acid was secreted into the cell medium during the incubation, it could not be assayed reliably by the hydroxysteroid dehydrogenase assay method, contrary to the reports of Anwer et al . 1975 . Biochem . Biophys . Res . Commun . 64: 603 and Gardner and Chenouda 1978 . J . Lipid Res . 19: 985. Am Rev Respir Dis, 1979 Sep, 120(3), 541 - 5 The effect of hyperoxia on migration of alveolar macrophages in vitro; Bowles AL et al.; There is in vitro evidence to support the notion that directed migration (chemotaxis) is involved in the recruitment of alveolar macrophages in vivo . Because O2 is widely used in the treatment of pulmonary diseases, we examined the effect of hyperoxia on migration of guinea pig alveolar macrophages in vitro . Migration was measured in blind-well chambers incubated in either room air or hyperoxia . N-formyl-methionyl-phenylalanine was used to stimulate random migration and to produce directed migration . Migration was quantified by counting the number of mononuclear cells per oil immersion field that had migrated completely through a polycarbonate filter with 5-micrometer pores . The average PO2 in the cell suspensions incubated in room air was 100 mm Hg . In the hyperoxic environments, the average PO2 at 1 h was 260 mm Hg, whereas at 2 and 3 h, it was 410 and 425 mm Hg, respectively . In 6 separate experiments, there was no significant difference between the mean response to N-formyl-methionyl phenylalanine in hyperoxia and in room air after 1 h of incubation . After 2 and 3h of incubation, however, the response in hyperoxia was significantly (P less than 0.002) lower than that in room air . The decreased response in hyperoxia did not appear to result from loss of viability of responding cells, diminished adherence of cells to the filters, loss of activity of N-formyl-methionyl phenylalanine exposed to high PO2, or failure of the cells to exhibit directed migration . Instead, it appeared that hyperoxia decreased the response of alveolar macrophages primarily by impairing random migration. Am J Physiol, 1979 Sep, 237(3), H348 - 52 Application of an isolated heart model to investigate blood-oxygen delivery; Rand PW et al.; To avoid the compensatory hemodynamic responses, which have limited interpretation of hemoglobin-oxygen affinity modifications in animal experimentation, an isolated blood-perfused rabbit heart model providing metabolic, functional, and vectorcardiographic measurements has been developed . Fixed-flow perfusions of unchanged or affinity-modified red blood cell suspensions were carried out to assess the benefits of high affinity during hypoxic hypoxia and of low affinity during posthypoxic recovery . Using fully saturated suspensions, the influence of affinity level during restricted flow and reperfusion was also studied . Higher myocardial oxygen consumption (MVO2) was associated with high-affinity blood during mild hypoxia and low-affinity blood during posthypoxic recovery . At low flows, heart rate and MVO2 tended to be lower in high-affinity perfusions, and to recover more completely during low-affinity reperfusions . Ventricular function, vectorcardiographic patterns, and lactate levels were affected by hypoxia and ischemia, but not by level of affinity . The relevance of these observations to the therapeutic potential of hemoglobin-oxygen affinity modification is discussed. Pflugers Arch, 1979 Sep, 381(3), 281 - 5 45Ca2+ uptake by dispersed pancreatic islet cells: effect of D-glucose and the calcium probe, chlorotetracycline; Sehlin J et al.; Uptake of 45Ca2+ was studied in dispersed pancreatic islet cells from non-inbred ob/ob-mice . Like whole islets the dispersed cells responded to 20 mM D-glucose with a markedly increased 45Ca2+-labeling of both the lanthanum-nondisplaceable and the lanthanum-displaceable calcium pools . The pronounced effect of D-glucose could not be reproduced with 3-O-methyl-D-glucose, L-glucose, D-mannose, L-leucine, or D-leucine; however, 45Ca2+ uptake was greater in the presence of L-leucine as compared with D-leucine . 45Ca2+ uptake by dispersed cells or whole islets was stimulated severalfold by 100 microM or more chlorotetracycline . At the concentration of only 10 microM, chlorotetracycline had no effect on whole islets and partially inhibited 45Ca2+ uptake by the dispersed cells . The ability of D-glucose to stimulate 45Ca2+ uptake by islets or dispersed cells remained in the presence of 10 microM chlorotetracycline . Islet cell suspensions apparently represent a valid model for studying how Ca2+ interacts with the cells . However, when using chlorotetracycline as fluorescent Ca2+ probe, attention must be paid to it potential ionophoric activity . At only 10 microM, the drug seems to monitor a peripheral pool of Ca2+, some of which may reside in normal transport channels. Exp Hematol, 1979 Sep, 7(8), 416 - 24 Engraftment of bone marrow transplants in W anemic mice measured by electronic determination of the red blood cell size profile; Wiktor-Jedrzejczak W et al.; Defective stem cells of WBB6F1-W/Wv mice produce macrocytic red blood cells (RBCs); stem cells of WBB6F1-+/+ mice produce normocytic RBCs . Utilization of the Coulter counter channelyzer permitted good dissociation between the size distribution of populations of +/+ and W/Wv RBCs . Peaks (mean cell volumes) for +/+ and W/Wv RBCs have been determined to be between the 30th and 40th channel and 50th and 60th channel, respectively . Variability of profiles for individual mice of both genotypes did not exceed the variability of separate determinations of the same cell suspension from a single mouse . Admixture (approximately 15%) of either type of erythrocytes could be quantitatively detected by this method . One week after transplant of 10(7) +/+ marrow cells into W/Wv recipients, 25% of donor type erythrocytes were detected . Eighteen days post-graft, concentration of +/- normocytes exceeded the concentration of macrocytes in the W/Wv recipients' circulation . Approximately 45 days post-transplant, the proportion of macrocytes decreased below the 10% detectable level . Calculation of the daily RBC production rate during repopulation and estimation of the number of RBCs produced by a single hematopoietic colony were determined . The RBC size profile was found to be a convenient method for studying the effect of implantation of W/Wv marrow into lethally irradiated +/+ mice . This method proved suitable for repetitive determination of the size population in individual transplanted mice. J Natl Cancer Inst, 1979 Sep, 63(3), 587 - 92 Relationship between monocytosis and T-lmphocyte function in human cancer; Wood GW et al.; This study was designed to: 1) determine the relative number of monocytes in mononuclear cell suspensions derived from the peripheral blood of cancer patients and 2) ascertain if any relationship existed between the numbers of monocytes in those cell suspensions and T-lymphocyte function . Monocytes were quantitated by morphology verified by phagocytosis of antibody-coated erythrocytes . A significant difference (P less than or equal to 0.01) existed between the number of monocytes in suspensions from normal individuals (26.2 +/- 4.3) and cancer patients (38.0 +/- 13.4) . The cancer patients were divided into 2 groups: 1) those who exhibited normal in vitro T-cell responses to phytohemagglutinin and 2) those in whom responses were significantly suppressed . The mean number of monocytes in suspensions from the cancer patient group with normal responses was 29 +/- 9, whereas that from the cancer patients with suppressed responses was 47 +/- 11, a highly significant difference (P less than or equal to 0.01) . Therefore, the study demonstrated two things: 1) Mononuclear cell suspensions derived from cancer patients exhibited significant monocytosis relative to those from normal individuals and 2) a strong correlation existed between monocytosis and suppressed T-lymphocyte function in vitro. J Immunol, 1979 Sep, 123(3), 1356 - 61 In vitro antibody response to influenza virus . I . T cell dependence of secondary response to hemagglutinin; Anders EM et al.; A good secondary IgG response to the hemagglutinin (HA) of influenza virus has been obtained in vitro in Marbrook-type cultures of influenza-primed mouse spleen cell suspensions stimulated with inactivated influenza virus . Anti-HA antibody was quantitated by a solid phase radioimmunoassay (RIA) by using purified HA as substrate . The T dependence of this secondary response was shown by depletion of T cells and reconstitution with a source of primed or unprimed T cells . The help given by T cells primed to the homologous virus was many times greater than that given by unprimed T cells, although the latter was significant . The system described will allow investigation of the specificity requirements of helper T cells engaged in the anti-HA response. J Lab Clin Med, 1979 Sep, 94(3), 414 - 20 A method for obtaining human bone marrow specimens enriched for myeloblasts and promyelocytes; Preisler HD et al.; A simple method has been developed for obtaining specimens of human marrow which are enriched for myeloblasts and promyelocytes . The erythrocytes are lysed, the marrow is incubated with iron particles, and the cells that phagocytize the iron are removed with a powerful magnet . The marrow is then subjected to a density-cut centrifugation using Ficol-Hypaque with a density of 1.084 gm/mm3 . The cells that do not enter the Ficol-Hypaque are removed from the surface and studied . The proportion of myeloblasts and promyelocytes in this subpopulation of cells exceeds 50% . Total recovery of these immature myeloid progenitor cells is 50% of that in the original marrow specimen . This method has been used for cell suspensions containing as many as 10(9) cells . Cells prepared using this method incorporate 3H-TdR, 3H-UR, and 3H-Leu at a higher rate than the unseparated specimens and have a cloning efficiency of 2.0% to 19.4% compared with 0.1% to 2.17% for the unseparated marrows. Blood, 1979 Sep, 54(3), 703 - 12 Hand-mirror cell leukemia associated with mental retardation: immunologic, chromosome, and morphological studies; Stern R et al.; Cycochemical, morphological, immunologic, and cytogenetic studies were carried out on hand-mirror cells (HMC) from a mentally retarded patient with a constitutional chromosome abnormality, 46,XX,r(21), and acute lymphoblastic leukemia . Scanning electron and differential interference contrast microscopy showed microspikes on the uropodia, but little evidence of cellular motility, despite formation and disappearance of individual uropodia in cell suspensions . The cells rosetted with sheep erythrocytes, suggesting T-cell origin . Cells derived from the bone marrow (80% HMC) showed a high degree of polyploidy (60%) and a bimodal chromosome number of 49 (49,XX,+10,-21, +3 rings) and 94 (6 no . 10, 3 no . 18, 2 no . 21 chromosomes, 3 ring chromosomes, plus 4 copies of each other chromosome). Z Naturforsch {C}, 1979 Sep-Oct, 34(9-10), 704 - 8 A model mechanism for the enzymatic synthesis of lupin alkaloids; Wink M et al.; A crude enzyme preparation obtained from cell suspension cultures of Lupinus polyphyllus catalyzes the pyruvate dependent conversion of cadaverine into the tetracyclic lupin alkaloids . As the first reaction product 17-oxosparteine could be identified by gas-liquid chromatography and mass spectroscopy . In some experiments sparteine was found additionally . A participation of diamine oxidase could be ruled out . The cadaverine-pyruvate transaminating enzyme system (17-oxosparteine synthase) catalyzes the formation of 17-oxosparteine from three cadaverine units without releasing free intermediates . These results are inconsistent with the hypothetical mechanism thus far formulated for the lupin alkaloid biosynthesis . A new enzymatic model mechanism is proposed regarding both the results of the enzymatic experiments and those of the in vivo tracer studies. Am J Physiol, 1979 Sep, 237(3), R132 - 8 Kinetics of bicarbonate/chloride exchange in dogfish erythrocytes; Obaid AL et al.; A stopped-flow rapid reaction apparatus was used to monitor changes in extracellular pH in dogfish (Mustelus canis) erythrocyte suspensions under conditions where dpH/dt was determined by the rate of HCO3-/Cl- exchange across the red cell membrane . Experiments were performed on erythrocytes suspended either in their own plasma or in elasmobranch Ringer solution over a range of temperatures from 5 to 35 degrees C . The exchange fluxes at 25 degrees C for red blood cells suspended in their own plasma (2.03 nmol/cm2-s) or in Ringer solution (2.00 nmol/cm2-s) are not significantly different and can be compared to those obtained under similar conditions for human red cell suspensions (0.910 nmol/cm2-s) . The flux for dogfish erythrocytes suspended in Ringer solution was reduced by 80% after exposure of the cells to SITS . An Arrhenius plot of the exchange rate constant yielded an activation energy of about 13.2 kcal/mol . We conclude that 1) plasma has no inhibitory effect of HCO3-/Cl- exchange across the dogfish erythrocyte membrane or on activity of intraerythrocyte carbonic anhydrase, 2) HCO3-/Cl- exchange probably occurs via the same mechanism in fish and mammalian erythrocytes, and 3) the conversion of plasms HCO3- to CO2 in dogfish can be catalyzed by intraerythrocyte carbonic anhydrase. Crit Care Med, 1979 Sep, 7(9), 391 - 5 Automated method for determination of oxygen equilibrium curves of red cell suspensions under controlled buffer conditions and its clinical applications; Asakura T; The accurate determination of the oxygen equilibrium curve (OEC) of whole blood or red cell suspensions requires special considerations to avoid the secondary effects due to lactate formation, removal of carbon dioxide, use of anticoagulants, etc . An automated apparatus has been constructed that can record the whole OEC of red cells within 20 min . The instrument also can be used to record the OEC of hemolysate . The pH, temperature, and PO2 are monitored constantly during the measurement . Using this instrument, the effects of pH, temperature, carbon dioxide, anticoagulants, and buffers on the OEC of normal fresh blood have been investigated . The OEC of normal blood, blood containing abnormal hemoglobin, or enzyme, were determined . The results were compared with those obtained using commercial OEC apparatuses. J Biol Chem, 1979 Aug 25, 254(16), 7885 - 94 Synthesis and secretion of corticotropins, melanotropins, and endorphins by rat intermediate pituitary cells; Mains RE et al.; The synthesis and secretion of various intermediate pituitary proteins was studied by using dispersed intermediate pituitary cell suspensions . Control studies indicated that the isolated cells were obtained in good yield and that after more than 24 h in culture the isolated cells continued to synthesize a collection of proteins similar to those found in freshly extracted intermediate pituitary tissue . Rat intermediate pituitary cells synthesized a molecule (Mr = 30,000; called 30K) that contained antigenic determinants for beta-endorphin, gamma-lipotropin, corticotropin (ACTH), and 16K fragment (the NH2-terminal region of mouse tumor cell pro-ACTH/endorphin) . This 30K molecule, two high molecular weight forms of ACTH(13K and 20K), and 16K fragment were all shown to be glycoproteins . Continuous labeling and pulse-chase incubations were used to define the intracellular biosynthetic processing of the 30K molecule . After a 15-min pulse incubation the 30K molecule was the only labeled protein containing antigenic determinants for beta-endorphin, gamma-lipotropin, ACTH, or 16K fragment . A beta-lipotropin-like molecule served as a biosynthetic intermediate in the production of proteins similar to beta-endorphin and gamma-lipotropin . Methionine-enkephalin and alpha-endorphin were not major products in the intermediate lobe cells . Molecules similar to alpha-melanocyte-stimulating hormone and corticotropin-like intermediate lobe peptide (ACTH(18-39)) were also derived from the same 30K molecule; 20K ACTH served as a biosynthetic intermediate in this conversion . In rat intermediate pituitary cells ACTH(1-39) was not a major final product of the intracellular biosynthetic processing of the 30K molecule . The 30K molecule also served as a precursor to a protein similar to mouse tumor cell 16K fragment and related smaller proteins . With rat intermediate pituitary cells, pulse-chase experiments utilizing {35S}methionine demonstrated almost quantitative conversion of the 30K precursor into labeled proteins similar to beta-endorphin and alpha-melanocyte-stimulating hormone . In the absence of added secretagogues, small amounts of all of the smaller proteins derived from the 30K precursor were secreted coordinately into the culture medium. Brain Res, 1979 Aug 3, 171(2), 225 - 37 Neurochemical and morphological studies of bulk isolated rat brain cells . II . Preparation of viable cerebral neurons which retain synaptic complexes; Huttner WB et al.; The bulk isolation from rat cerebral cortex of viable neurons retaining synaptic complexes is described . The basis of this procedure is to dissociate the neurons in situ from the surrounding glial cells . The glial structures that are normally adjacent to the neuronal cell body and to the proximal parts of the neuronal processes are largely destroyed by perfusion of the brain under special conditions . The most important of these conditions was found to be a hyperosmolar concentration of hexoses in the perfusion medium . In addition, the presence of collagenase and hyaluronidase in the perfusion medium and specific perfusate flow characteristics were required to produce the structural changes throughout the brain tissue . When the perfused brain was further dissociated into a cell suspension by mincing and sieving, isolated neurons were obtained, the majority of which retained the proximal parts of their processes . A novel feature of these neurons was the retention of synaptic boutons on the plasma membrane . Presynaptic terminals with mitochondria and vesicles as well as pre- and postsynaptic membranes and densities were observed on the isolated neurons . The neurons were fractionated to 90--95% purity using discontinuous Ficoll density gradient centrifugation with a liquid fluorocarbon as cushion . Highly purified, viable cerebral neurons retaining synaptic complexes are thus available in bulk for neurobiological studies. Scand J Haematol, 1979 Aug, 23(2), 129 - 40 Effect of human serum from patients with haematological disorders on mouse pluripotent haemopoietic stem cells (CFU-S); Sawada U et al.; Sera from 25 individuals (9 healthy subjects, 6 patients with systemic lupus erythematosus, and 10 patients with a variety of haematological disorders) were tested for anti-CFU-S by incubating suspensions of mouse marrow cells with serum and then assaying the cell suspensions for their capacity to form spleen nodules in lethally irradiated mice . The sera from 2 patients (1 with a preleukaemic disorder and the other with a malignant 'histio'-lymphoproliferative neoplasm) had anti-CFU-S activity which behaved like antibodies, rather than like the non-specific cytotoxic activity found in various sera for heterologous tissues, which in the past has been attributed to 'natural antibodies' and which, in recent years, has, in some cases, been found to be related to activation of the alternative pathway of complement . The anti-mouse tissue activity in the sera of the 2 patients may be related either to cross-reactivities between certain mouse and human tissues or to cross-reactivities between exogenous agents such as bacteria and mouse tissues. J Pharm Sci, 1979 Aug, 68(8), 1022 - 4 Myoinositol uptake by rat hepatocytes in vitro; Chen CP et al.; Myoinositol uptake by rat hepatocytes in vitro was studied . Adult rat hepatocytes were prepared by digestion of the perfused liver with collagenase . Cell suspensions were incubated with tritium-labeled myoinositol in pH 7.4 Krebs bicarbonate solution containing 1% gelatin at 37 degrees . 14C-Carbon-labeled polyethylene glycol was used as a marker of adherent extracellular fluid volume . Myoinositol uptake was demonstrable after 5 min of incubation, but no intracellular concentration in excess of that in the incubation medium was observed after 60 min of incubation . Uptake saturation over a wide myoinositol concentration range could not be demonstrated . Neither the omission of sodium ions nor the inclusion of ouabain suppressed the distribution ratio significantly . Metabolic inhibitors and lower temperatures also show