Culture medium

J Immunol, 1981 Feb, 126(2), 540 - 4
Fate of a common acute lymphoblastic leukemia antigen during modulation by monoclonal antibody; Pesando JM et al.; Modulation of a human common acute lymphoblastic leukemia antigen (CALLA) by specific monoclonal antibody (J5) has been studied with the immune precipitation method to identify radiolabeled antigen . Surfaces of leukemic cells have been labeled using 125I both before and after modulation by J5 antibody for different time intervals . Leukemic cells have also been metabolically labeled with 35S-methionine before modulation . These studies indicate that the 100,000-dalton glycoprotein expressing CALLA (gp 100-CALLA) cannot be detected in cells that were modulated with J5 antibody before surface labeling but that it is easily detectable in cells that were surface labeled before modulation for 10 hr . At later time points, gp 100-CALLA is selectively lost from cells that were surface labeled before modulation . Gp 100-CALLA is not detected in the supernatants from cultures of these modulated cells . We conclude that gp 100-CALLA is rapidly internalized during modulation and that CALLA is degraded . Gp 100-CALLA is not shed into the culture media, nor does it remain on the cell surface in an altered form . Incubation of leukemic cells with antisera to beta2-microglobulin or IgM does not affect the expression of gp 100-CALLA.

Infect Immun, 1981 Feb, 31(2), 704 - 11
Effect of herpes simplex virus infection on murine antibody-dependent cellular cytotoxicity and natural killer cytotoxicity; Kohl S et al.; Mice intraperitoneally inoculated with a sublethal dose of herpes simplex virus (HSV) produced immunoglobulin G antibody-dependent cellular cytotoxicity (ADCC) and radioimmunoassay (RIA) antibody as early as 3 days after infection . There was a rise in natural killer cytotoxicity (NKC) to infected and uninfected target cells 1 to 3 days postinfection mediated by nonadherent peritoneal cells (PC) in mice inoculated with HSV, but also with other substances commonly used in tissue culture media . HSV caused the highest and most consistent increase in NKC . PC-NKC, as ADCC, was inhibited by latex and silica, both macrophage inhibitors . PC-ADCC markedly declined 3 to 8 days after HSV inoculation . This was not due to a soluble or cellular suppressor factor, was not reversed by incubation or trypsin treatment of PC, was not associated with a change in PC Fc receptors, adherence, or acridine orange staining characteristics, and could not be induced by inactivated HSV . In vitro inoculation of PC with HSV similarly caused a reduction in the ability of PC to mediate ADCC to HSV-infected target cells . These data demonstrate the complex stimulatory and inhibitory interactions between virus and host defense mechanisms.

Chromosoma, 1981, 84(3), 365 - 72
Cell-specific variation of X chromosome replication in the Syrian hamster (Mesocricetus auratus); Cawood AH; The sub-division of S-phase in Syrian hamsters, on the basis of Brd/U/Hoechst 33258/Giemsa banding, has allowed a quantitative comparison of the replication of individual chromosome bands within defined subphases of S . This analysis has shown that in hamsters, as has been reported in humans, there are distinct patterns of early replication in vitro in the early X, the late X in fibroblasts, and the late X in lymphocytes . In addition, it has been possible to show that, although the pattern of replication of the late X in fibroblasts differs from that in lymphocytes, the time in S at which bands first appear on this chromosome is the same in the two cell types . No significant heterogeneity can be ascribed to differences between individuals, adult or embryonic sources, culture media, or time of exposure to BrdU . The absence of any detectable heterogeneity in the replication band frequencies in autosomal heterochromatic arms suggests that the cell-specific variability of the late-replicating X is a feature of facultative rather than constitutive heterochromatin.

Histochemistry, 1981, 73(1), 137 - 50
{The histochemistry of human endosalpingeal ciliated cells (author's transl)}; Kugler P; The human endosalpingeal ciliated cells are not a homogeneous cell population . They can be distinguished into mitochondria prominent and normal cells . The morphological appearance of ciliated cells was studied in organ culture using different hormones and hormone combinations in the culture media . Histological (HE, semithin sections) and histochemical methods (PAS, Alcian Blue, SDH, LDH, ATPase, 5'-Nucleotidase, acid and alkaline Phosphatase) were applied . Those mitochondria prominent ciliated cells which are seen in the native endosalpinx can in vitro also be determined mainly under the influence of steroid hormones (hydrocortisone and progesterone) . In hormone free incubation media the apices of ciliated cells are separated . This will happen in hormone containing media, too, but with delay, however . Some results are in agreement with the possible transformation from ciliated cells to secretory cells.

Folia Microbiol (Praha), 1981, 26(5), 408 - 12
Production and purification of Penicillium citreoviride toxin and its effect on TPP-dependent liver transketolase; Datta SC et al.; The production isolation and purification of a yellow mycotoxin from Penicillium citreoviride NRRL 2579 in different culture media was described . When injected subcutaneously to albino rats it alters the kinetic pattern of transketolase (EC 2.2.1.1) in liver in vivo in a competitive manner . In vitro, the inhibition is noncompetitive in nature . However, addition of thiamine diphosphate (TPP) to the in vitro system relieved the inhibitory effect . These findings suggested a relationship between citreoviridin-induced beriberi and the probable antithiamine effect of the toxin.

J Immunol Methods, 1981, 44(1), 3 - 14
Human lymphocyte transformation induced by mitogens and antigens in a serum-free tissue culture system; Needleman BW et al.; The use of serum to supplement lymphocyte tissue culture media introduces uncontrolled variables; different serum sources, lots and concentrations can produce variability in experimental results, serum can stimulate or inhibit lymphocytes, and components of serum can react with substances whose effects on lymphocytes are being studied . To avoid these problems, we studied the ability of human peripheral mononuclear cells to survive and to respond to stimulation in an entirely synthetic medium, RPMI-1640 supplemented with L-glutamine, gentamicin, HEPES buffer and magnesium . Optimal cell concentration in this serum-free RPMI-1640 was 2.5 x 10(6) cells/ml, whereas optimal cell concentration in serum containing RPMI-1640 was 1 x 10(6) cells/ml . In this serum-free RPMI-1640, 50% of the cellular input was recovered as viable cells after 7 days of culture, which was similar to results in serum containing RPMI-1640 . Mononuclear cell transformation transformation was induced by phytohemagglutinin, concanavalin A, pokeweed mitogen, streptolysin O and candida . Optimal doses of stimulants and the kinetics of the responses were similar in serum-free and serum containing RPMI-1640 . This system can be used to avoid the problems inherent in systems which supplement tissue culture media with serum.

Arch Virol, 1981, 67(1), 31 - 43
The detection of influenza A virus antigens in cultured cells by enzyme-linked immunosorbent assay; Watanabe H et al.; An enzyme-linked immunosorbent assay (ELISA) was employed to investigate the expression of influenza A/Hong Kong/68 (H3N3) virus structural proteins on the surface of infected MDCK cells, and to detect viral antigens in culture media and cell extracts . Infected cells were fixed with 0.1 per cent glutaraldehyde before being examined for the presence of cell-surface antigens . Viral antigens were first observed on the surface of cells 4 hours after infection and reached a maximum 10-12 hours after infection, when measured by haemadsorption with chicken erythrocytes and by ELISA and immunofluorescence with hyperimmune antiserum to Hong Kong virus . A good correlation was found between the three assay systems . The presence of individual virion structural proteins on the cell surface was determined by ELISA using specific antibodies purified by differential affinity chromatography . Either or both or the internal matrix and nucleoprotein antigens were expressed from 2 to 6 hours after infection, with maximum expression after 2 hours, and the strain-specific and common antigenic determinants of haemagglutinin were observed on the cell surface from 4 hours after infection, and reached a maximum 8 to 10 hours after infection . Low levels of neuraminidase were detected between 4 and 8 hours after infection . Culture media and cell extracts were titrated by infectivity and haemagglutination assays, and by ELISA . Titres obtained from the culture media showed a close correlation between the three assay methods, with peak titres being attained 24 hours after infection . Viral antigens were first observed in cell extracts by ELISA 4 hours after infection, and infectious virions and haemagglutinin 2 hours later, but whereas maximum titres of infectious virus and haemagglutinin were found 10 hours after infection, the ELISA titre continued to rise until 24 hours after infection, which suggested that virus structural proteins were being accumulated in the cells after most of the progeny virions had been released . The results are discussed in terms of the potential use of ELISA in rapid virus diagnosis.

Med Interne, 1981 Jan-Mar, 19(1), 73 - 7
The relationship between air-borne fungal spores and Dermatophagoides pteronyssinus in the house dust; Chirila M et al.; The prevalence of mould genera and their association with the house dust mite (Dermatophagoides pteronyssinus) were investigated in the homes of 192 asthmatic patients and 125 healthy people in a Romanian town with a relatively high degree of humidity . The dominant moulds which developed in the culture media exposed on the bedroom floors were Cladosporium, Penicillium and Aspergillus . Living Dermatophagoides pt . and their eggs were present mostly in the colonies of Cladosporium; in those of Penicillium the house dust mites were dead and disintegrated . It is concluded that allergy to Dermatophagoides pt . must not be connected to house dust as such but to the presence in the dust of certain mould spores.

Cancer Chemother Pharmacol, 1981, 6(3), 205 - 10
Methodologic problems in clonogenic assays of spontaneous human tumors; MacKintosh FR et al.; Colony formation in soft agar was used to investigate growth properties and drug sensitivity in 102 tumor specimens from 91 patients . Sufficient colony growth for sensitivity testing with various drugs was obtained in 36 of 67 specimens (54%) with adequate cell yield and pathologically documented malignancy . Room temperature (20-24 degrees C) is superior to both 4 degrees C and 37 degrees C for 12-36 h storage and transport of malignant effusions . By contrast, fine mincing in sterile saline or balanced salt solution, and refrigerated storage (4 degrees C) appear optimal in experiments with three solid tumors . The use of buffered NH4Cl to lyse red blood cells markedly reduced plating efficiencies, and also reduced the percentage of tumors in which drug sensitivities could be tested from 64% to 38% . Several combinations of potential growth factors and culture media have been tested . Insulin enhanced plating efficiency (PE) in all six adenocarcinomas tested . Drug sensitivity of tumors was not affected by varying plating efficiency up to five-fold in two tumors . In eleven cases tumor cells were exposed to combinations of two or more drugs, and results assessed for evidence of drug interactions . In almost all cases, these two-drug combinations produced additive cell killing than either antagonistic or greater-than-additive effects.U

Clin Orthop, 1981 Jan-Feb, (154), 234 - 48
Bone-forming and bone-resorbing cell lines derived from bone marrow in tissue culture; Hirano H et al.; The differentiation of connective tissue outgrowths of adult bone marrow in response to the organic matrix of bone was observed in tissue culture by correlated histologic, electron microscopic, biochemical, and radioisotope-labeling methods . On a substratum of bone matrix gelatin, myelogenous cells degenerate and disappear while stromal and perivascular cells proliferate and differentiate into mesenchymal-type, monocytoid, and giant cells . From primary cultures of bone marrow cells on bone matrix, cartilage differentiates in isolated areas but only in small islets . With continuous subculture through 25 generations, the proportions of two functionally different populations of chondrogenetic and matrix-resorbing cells gradually emerge . Up to the time of the ninth generation of subculture, the chondrogenetic population predominates . After the 14th to the 25th generation, clones of matrix-resorbing large monocytoid cells predominate and rapidly digest the matrix substratum . Measurements of 35S uptake demonstrate that control cultures of muscle-derived mesenchymal-type cells produce about twice as much cartilage as marrow-derived mesenchymal-type cells . A decline in the chondrogenetic cell population and corresponding rise in the matrix-resorbing cell population is demonstrable by a progressive increase in the quantity of hydroxyproline-containing peptides in the culture medium . This decline is not attributable to conditions in culture because there was progressive loss of chondrogenetic activity of the eighth and 20th passage even when the cells were transplanted in diffusion chambers back into an isologous host . The problem is how to account for the competence of marrow stromal cells to differentiate into bone without bone matrix in vivo but not in vitro . Mixed cultures of muscle and marrow outgrowths produce only half as much cartilage (measured by 36S-uptake/microgramDNA in the system) as muscle outgrowths alone . These observations suggest that a bone marrow-derived matrix-resorbing cell population, by some unknown mechanism, inhibits proliferation of cartilage-bone precursor cell populations . The nature of the inhibition requires investigation by detailed biochemical analyses of marrow cell culture media and chemical extracts of whole bone marrow tissue.

J Cancer Res Clin Oncol, 1981, 102(1), 49 - 55
Human erythroid cell lines derived from a patient with acute erythremia; Okano H et al.; Three continuous human cell lines, designated KMOE, derived from a patient with acute erythremia (Di Guglielmo's disease) are reported . The cell lines are the cultures of (1) bone marrow cells, (2) peripheral blood cells, and (3) cells from a tumor developed into an athymic nude mouse after transplantation of the cultured bone marrow cells . Cells of all three lines show morphology of immature erythroblast and have i(17q) marker chromosome . They are negative for both Philadelphia chromosome and Epstein-Barr virus nuclear antigen . Although all KMOE cells in suspension culture are benzidine-negative, benzidine-positive cells are found within colonies formed in semi-solid culture media . The relative number of colonies with benzidine-positive cells is increased when sodium butyrate is added to the culture.

Pharmacol Ther Dent, 1981, 6(1-2), 35 - 43
The effect of dental plaque grown in the presence of xylitol or sucrose on bone resorption in vitro; Tenovuo J et al.; Bone culture was used as an experimental model in studying the ability of dental plaque grown in the presence of xylitol or sucrose to induce bone resorption . Plaque samples were collected in young adults after six or ten days with no oral hygiene and with frequent use of xylitol- or sucrose-sweetened chewing gum . The rate of resorption was assayed by measuring the release of both acid phosphatase and 45Ca from bones into culture media during a three-day incubation period . Sucrose-induced plaque collected after ten days increased both of these indicators of bone resorption, while xylitol plaque decreased or had no effect under identical conditions . Xylitol consumption induced a marked increase in acid phosphatase activity of dental plaque - a phenomenon which would appear to be unrelated to bone resorption . The results suggest that the inflammatory potential of dental plaque may be reduced during xylitol consumption as compared to sucrose consumption.

Proc Natl Acad Sci U S A, 1981 Jan, 78(1), 162 - 6
Isolation of functional human coagulation factor V by using a hybridoma antibody; Katzmann JA et al.; Spleen cells obtained from mice immunized with partially purified human coagulation Factor V were fused with NS-1 mouse myeloma cells, and hybrids were selected . Culture media were screened for anti-Factor V activity, and an antibody-positive clone was obtained and passaged as an ascites tumor in mice . The ascitic fluid from the hybridoma-bearing mouse could be diluted 1:10(6) before losing reactivity in an anti-Factor V radioimmunoassay . When immobilized on agarose, the monoclonal antibody quantitatively removed Factor V activity from human plasma . Factor V activity could be eluted with 1.2 M NaCl at pH 6.5 . Homogeneous Factor V was isolated by chromatography of barium citrate-adsorbed, polyethylene glycol 6000 precipitated plasma on the antibody column followed by chromatography on phenyl-Sepharose . The isolated Factor V exhibited a single band upon gel electrophoresis in sodium dodecyl sulfate with an apparent Mr comparable to that of bovine Factor V (330,000) . Upon exposure to thrombin, the activity of Factor V increased 53-fold when measured in Factor V-deficient plasma . This increased activity was associated with discrete proteolytic cleavages of the parent molecule.

Acta Physiol Lat Am, 1981, 31(3), 199 - 206
Immunogenicity of Trypanosoma cruzi in different animal species; Rubiolo ER; The immunogenicity of two fractions (1 500 F and 10 000 F) from epimastigotes of Trypanosoma cruzi as well as the supernatant from culture media (SF) were studied using hens, rabbits and opossums . For comparative purposes, sera from individuals with chronic Chagas' disease were also used . A similar, positive response was obtained for the fractions in all the animal species studied using indirect hemagglutination test . Supernatants from culture media were the least immunogenic . By double immunodiffusion test, it was possible to detect a positive response to a different number as well as to different antigens in the three animal species, but there was response to a common antigen by all the different animal species . The common antigen called here major, was present in all the fractions assayed . Human sera from individuals chronically infected showed a variable response . When assayed by double immunodiffusion technique, the major antigen could be detected in just a few samples.

Mol Cell Endocrinol, 1981 Jan, 21(1), 43 - 9
Inhibitory effect of gonadotropin releasing hormone upon cultured testicular cells; Hsueh AJ et al.; The direct inhibitory of gonadotropin-releasing hormone (GnRH) upon testicular androgen production was studied in cultured testicular cells . Enzyme-dispersed testicular cells from immature hypophysectomized rats were cultured for 6 days in a serum-free medium with or without various hormones . Culture media were changed every 2 days and media concentration of a androgens was measured by radioimmunoassay . The testis cultures from immature rats secrete predominately 5 alpha-androstane-3 alpha, 17 beta-diol (A-diol) and androsterone but negligible amounts of androstenedione, testosterone and dihydrotestosterone . Incubation of testicular cells with hCG and FSH increased A-diol and androsterone production as compared to control cultures . In contrast, treatment with GnRH or a GnRH agonist (des-Gly10, D-Leu6 (N alpha Me)Leu7, Pro9NHEt-GnRH) inhibited the gonadotropin effect in a dose-dependent manner; 10(-8) M GnRH inhibited A-diol production by approximately 20% on day 4 and 6 of culture whereas 10(-6) M GnRH inhibited A-diol production by greater than 80% . The observed effect of GnRH upon testicular cells in primary culture demonstrates that the hypothalamic peptide directly inhibits testicular steroidogenesis.

Connect Tissue Res, 1981, 9(1), 19 - 24
Connective tissue activating macromolecules in macrophage culture medium; Jalkanen M; Fractions isolated by isoelectric focusing (pI values 4.6-5.3) from macrophage culture media stimulated fibroblast proliferation in vitro and increased the synthesis of sulfated glycosaminoglycans (sGAGs) . Cationic fractions from macrophage culture media isolated by the same procedure (pI 10.2-10.4) enhanced secretion of collagen by fibroblasts . Two dimensional electrophoresis of non-fractionated 3H-lysine labeled macrophage media only showed incorporation of label into acidic proteins . The fractions from the pI area which increased DNA and GAG synthesis by fibroblast cultures contained three major labeled proteins with molecular weights of 15 K, 25 K and 135 K.

Rheumatol Int, 1981, 1(1), 11 - 6
The production in culture of metalloproteinases and an inhibitor by joint tissues from normal rabbits, and from rabbits with a model arthritis . I . Synovium; Cambray GJ et al.; During 5 days of culture, explants of normal rabbit synovium produced no active collagenase, negligible latent collagenase, but significant levels of free collagenase inhibitor . Synovium from joints exhibiting a proliferative arthritis produced greatly elevated levels of collagenase; the appearance of active enzyme in the medium during the second day of culture was associated with the disappearance of free inhibitor . Enzyme levels in the media correlated well with the arthritic status of joints, when explants were prepared up to 10 weeks after the induction of the model arthritis . Synovium from the contralateral joints of rabbits with unilaterally induced arthritis produced no active collagenase, but approximately one-third as much latent collagenase as found with arthritic joints . Enzymatic activities against gelatin and cartilage proteoglycan substrates were demonstrated in synovial culture media in addition to collagenolytic activity . Gel filtration showed that these activities were not due to a single enzyme, and further characterisation confirmed that the enzymes were metalloproteinases . The results are considered in the light of published data, and the involvement of metalloproteinases and their specific inhibitor in the development of arthritic lesions is discussed.

Biochim Biophys Acta, 1980 Dec 15, 633(3), 410 - 21
Effect of RNA and protein synthesis inhibitors on the release of inflammatory mediators by macrophages responding to phorbol myristate acetate; Bonney RJ et al.; The interaction of phorbol myristate acetate with resident populations of mouse peritoneal macrophages causes an increased release of arachidonic acid followed by increased synthesis and secretion of prostaglandin E2 and 6-keto-prostaglandin F1 alpha . In addition, phorbol myristate acetate causes the selective release of lysosomal acid hydrolases from resident and elicited macrophages . These effects of phorbol myristate acetate on macrophages do not cause lactate dehydrogenase to leak into the culture media . The phorbol myristate acetate-induced release of arachidonic acid and increased synthesis and secretion of prostaglandins by macrophages can be inhibited by RNA and protein synthesis inhibitors, whereas the release of lysosomal hydrolases is unaffected . 0.1 microgram/ml actinomycin D blocked the increased prostaglandin production due to this inflammatory agent by more than 80%, and 3 microgram/ml cycloheximide blocked prostaglandin production by 78% . Similar results with these metabolic inhibitors were found with another stimulator of prostaglandin production, zymosan . However, these inhibitors do not interfere with lysosomal hydrolase releases caused by zymosan or phorbol myristate acetate . It appears that one of the results of the interaction of macrophages with inflammatory stimuli is the synthesis of a rapidly turning-over protein which regulates the production of prostaglandins . It is also clear that the secretion of prostaglandins and lysosomal hydrolases are independently regulated.

Biochim Biophys Acta, 1980 Dec 5, 620(3), 418 - 28
Cyclooxygenase products of arachidonic acid metabolism by mouse bone in organ culture; Voelkel EF et al.; The products of endogenous and exogenous arachidonic acid metabolism via the cyclooxygenase pathway in mouse bone in organ culture were identified and quantitated by the use of high performance liquid chromatography and radioimmunoassay . Production of prostaglandins E2, F2 alpha, and I2 from endogenous substrate was stimulated by incubation of bone with epidermal growth factor and the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate . Addition of arachidonic acid to the culture medium not only resulted in the accumulation of prostaglandins E2, F2 alpha, and I2 but also thromboxane . Bone metabolized prostaglandins E2 and F2 alpha to their respective 13,14-dihydro-15-keto-derivatives . Prostaglandin I2, measured as 6-keto-prostaglandin F1 alpha was synthesized by bone, and metabolic products of prostaglandin I2 or 6-keto-prostaglandin F1 alpha, either 6,15-diketo-prostaglandin F1 alpha or 13,14-dihydro-6,15-diketo-prostaglandin F1 alpha, were also detected in the culture media . Formation of cyclooxygenase products of endogenous and exogenous arachidonic acid metabolism (both basal and stimulated) and bone resorption were inhibited by indomethacin . Bone as a tissue responded biochemically not only to exogenous prostaglandins and agents that enhance endogenous prostaglandin production but also to exogenous arachidonic acid by biosynthesis of prostaglandins, prostacyclin and thromboxane . Furthermore, bone metabolized these cyclooxygenase products to their more stable metabolites.

Early Hum Dev, 1980 Dec, 4(4), 337 - 45
Human amniotic fluid contains fibroblast-pneumonocyte factor; Seybold WD et al.; We have studied 41 samples of human amniotic fluid collected at 27-40 wk gestation . After 'stripping' with Florisil, most fluids studied stimulated {3H} choline incorporation into saturated phosphatidylcholine by human fetal alveolar Type II cells . The activity declined with advancing gestation and was inversely related to cell number following incubation . Following acid-ethanol extraction, activity eluted from Sephadex G75 columns in 1 M acetic acid with a Kd of approximately equal to 0.4-0.6, and was greater at earlier gestations (27-31 wk) than at later periods (32-36 or 37-40 wk) . In contrast, chromatography at neutral pH without prior extraction resulted in retarded elution . The active material was shown to be heat-stable . Thus, the biochemical features of activity recovered from human amniotic fluid are identical to fibroblast-pneumonocyte factor recovered from supernatant culture media of cortisol-treated human fetal lung fibroblasts.

Jpn J Exp Med, 1980 Dec, 50(6), 407 - 14
Analyses of some mite antigens found in house dust and in food material; Yago A et al.; Antigen analyses by micro-Ouchterlony, immunoelectrophoresis and passive hemagglutination tests were carried out on extracts from each of the following cultured mites: Dermatophogoides farinae, Dermatophogoides pteronyssinus, Tyrophagus putrescentiae, Aleuroglyphus ovatus and Carpoglyphus lactis; and on an extract from the culture medium for each mite . Antisera were raised against each extract or each medium in rabbits or in guinea pigs . Up to 5 precipitin lines by micro-Ouchterlony test and up to 8 precipitin arcs by immunoelectrophoresis were detected between an extract and its homologous (corresponding) antiserum . Some of the lines or arcs were seen consistently to be common to all species tested . The absorption of common antigens in an extract by heterologous (noncorresponding) antiserum was employed to find the antigens specific to D . farinae and D . pteronyssinus . By these tests, at least 3 species-specific antigens for D . farinae and 2 for D . pteronyssinus were found . In passive hemagglutination tests, higher titers were obtained mainly in homologous systems, although weak cross reaction was observed in every system . Culture media did not participate in any reaction combinations used here and did not interfere with the analyses.

Prostaglandins, 1980 Dec, 20(6), 1075 - 87
The prostaglandins of articular cartilage . I . Correlates of prostaglandin activity in a chondrocyte culture system; Copeland M et al.; Suspensions of aggregated chondrocytes display active prostaglandin (PG) production . Radioimmunoassay of culture media and thin layer chromatographic analysis suggests that PGE2 is the primary PG synthesized . In order of decreasing concentration, the following PG were tentatively identified; PGE greater than PGI greater than PGA + PGB greater than or equal to PGF1+2 greater than TxB . An inverse logarithmic relationship was identified between PG synthesis and cells cultured at densities of 1.5 to 7.5 x 10(6) cells/ml . Little or no change in the PG distribution profile was seen at these high cell densities . Maximum PG synthesis was attained after 36 hours of incubation with persistence of high synthetic levels up to 48 hours . PGE2 production measured at various post-isolation intervals indicated an initial high rate of synthesis during the first 4 hours which decreased with time up to 24 hours . Cartilage explant organ cultures demonstrated a similar level of PG synthesis suggesting minimal effect of matrix on cellular PG production . Indomethacin (5 microgram/ml) inhibited PG synthesis by 70% within 4 hours and 85% after 24 hours of exposure . Arachidonic acid supplementation (10 microM) stimulated PG synthesis by 300%.

J Clin Endocrinol Metab, 1980 Dec, 51(6), 1432 - 6
Pregnancy-specific beta-1-glycoprotein (SP1) in cultured amniotic fluid cells; Heikinheimo M et al.; The synthesis of pregnancy-specific beta-1-glycoprotein (SP1) was studied in amniotic fluid cell cultures using RIA, immunoperoxidase, and immunofluorescence techniques . SP1 was found by RIA in all 11 sonicates and in 21 of 26 culture media . The SP1-immunoreactive material was immunologically similar to maternal serum SP1 . Immunoperoxidase and indirect immunofluorescence staining were positive in large cells identified as epithelial amniotic cells by labeling with antikeratin antibodies . Fibroblast-like cells were occasionally found in cultures, but they did not contain demonstrable amounts of SP1 . The physiological significance of the findings presented remains unclear.

Cancer Res, 1980 Dec, 40(12), 4722 - 7
Specific effects of fibronectin-releasing peptides on the extracellular matrices of cultured human fibroblasts; Keski-Oja J et al.; Fibronectin-releasing activity has recently been found in concentrated serum-free culture media of the established human fibrosarcoma cell line, 8387 . We used these M.W . 10,000 fibronectin-releasing polypeptides (10K peptides) to study their effects on the cell-free matrices of cultured diploid human lung fibroblasts . Metabolically labeled cultures of fibroblasts were extracted with sodium deoxycholate and hypotonic buffer to prepare the matrices . The isolated matrices contained fibronectin and procollagen as their major radiolabeled proteins, and some as yet unidentified polypeptides were also detected . the matrices that were attached on the culture dishes were exposed to increasing concentrations of the 10K peptides in serum-free medium, and the changes in the radiolabeled polypeptides were studied . As a result of this treatment, there was a massive release of both fibronectin and procollagen from the matrices . No major cleavages of either released protein could be seen . After digestion of the matrix-associated collagen with collagenase, the 10K peptides released only fibronectin . Collagenase treatment, on the contrary, did not affect matrix-associated fibronectin . On the other hand, a M.W . 66,000 matrix-associated protein was constantly cleaved to a M.W . 62,000 form that remained in the matrix as a result of the incubation of the matrices with the 10K peptides . The 10K peptides did not affect the other radiolabeled polypeptides present in the matrix . the results indicate that the fibronectin-releasing peptide behaves as a specific protease on the matrices of cultured human fibroblasts.

Br J Vener Dis, 1980 Dec, 56(6), 390 - 3
Assessment of transport and isolation methods for gonococci; Taylor E et al.; Urethral discharge from men was diluted to give heavy and light inocula and cultured on seven different solid culture media, including two transport/isolation media, or held in three types of semi-solid transport medium for varying periods and then cultured . The amount of growth was quantitated and the performance of the different systems compared . Fresh non-selective media were best, with up to two failures in 254 cultures on each medium . With selective media there were 9-23 failures with heavy inocula and 22-47 failures with dilute inocula . For Transgrow or the Jembec system incubation before holding at ambient temperature was better than holding followed by incubation . Transport media yielded good results if cultures were set up within six hours; only minor losses occurred after 24 hours.

Cancer Lett, 1980 Dec, 11(2), 81 - 7
Formation of metastasis by human breast carcinoma cells (MCF-7) in nude mice; Shafie SM et al.; MCF-7 cells, a human breast carcinoma line, forms tumors when injected into athymic nude mice . These tumors are able to metastasize to lungs, liver and spleen . 17 beta-estradiol treatment increases both the growth rate and frequency of metastases . Castration or diabetes prevents metastasis formation, but treatment with estrogen or insulin restores the metastasizing capacity . MCF-7 cells secrete into the culture media collagenases able to lyse types I and IV collagens . Estrogen or insulin addition to the culture enhances collagenase production . Attention is called to the coexistence of enhancement in collagenase production and metastasis formation.

Zh Mikrobiol Epidemiol Immunobiol, 1980 Dec, (12), 57 - 60
{Modification of the agarose drop micromethod for studying mouse macrophage migration inhibition factor}; Lebedev VS; A modification of the drop agarose micromethod for the study of the factor inhibiting the migration of mouse macrophages is proposed . The syngeneic system was used instead of type RPMI-1640 complex culture media and calf embryonal serum, commonly used in the work with mouse macrophages . The production of the factor inhibiting the migration of macrophages by sensitized lymphocytes from mice of different strains has been shown to occur in vitro under the influence of pertussis corpuscular antigen . The method of obtaining supernatant containing macrophage migration inhibiting factor, from immune mouse lymphocyte culture is proposed . This supernatant may be used for, the analysis of the above-mentioned factor and other lymphokins.

Acta Obstet Gynaecol Jpn, 1980 Dec, 32(12), 1937 - 44
{Establishment of persistent cytomegalovirus infection in primary cultured trophoblastic cells (author's transl)}; Yabuki Y; The primary cultured cells (Ch) derived from chorionic villi were infected with human cytomegalovirus (CMV) . The main results obtained are as follows . 1) The primary cultured trophoblastic cell (Ch) infected with CMV have been maintained in the state of CMV persistent infection for over 6 months . These cells (Ch/CMV) were the CMV-carrier cells in a balance between the cytolysis by CMV-specific cytopathic effect (CPE) and the growth of uninfected cells, showing the CMV persistent infection at the cell population levels . 2) The Ch/CMV cells have persistently released infectious CMV with titers ranging from 1.6 X 10(2) to 1.2 X 10(4) PFU/ml into the culture fluid . CMV-specific antigens were detectable by immunofluorescent antibody staining of the virus releasing cells . 3) These Ch/CMV cells still maintained an ability to secret estrone and estradiol . When these abilities lowered in the successive cultures, the cells encountered the cytolysis by CMV release . 4) Infectious titer of CMV released from Ch/CMV cells reduced by the addition of estrogen in the culture media . In this case, however CMV-specific CPE occurred with a similar pattern as the control one lacked an addition of estrogen . 5) It was suggested that estrogen synthesis in the cells may play an important role in an establishment or a maintenance of CMV persistent infection.

Circ Res, 1980 Nov, 47(5), 770 - 5
Trophic effect of norepinephrine on the rat portal vein in organ culture; Abel PW et al.; Rat portal veins were maintained in organ culture to study the development of characteristic denervation changes and a possible trophic effect of the neurotransmitter norepinephrine (NE) . Vessels maintained in organ culture for 2 days showed supersensitivity to NE and Ba2+, a more rapid rate of relaxation from a Ba2+ contracture, and partial depolarization of the myovascular cells . All of these changes except the quicker relaxation from Ba2+ contracture could be prevented by incubating the preparations in a NE-containing medium . This evidence suggests that functional changes in vascular muscle cells are caused by the removal of a tropic influence of NE, but can be prevented by NE replacement . However, the failure of NE in the culture media to prevent the increased rate of relaxation from Ba2+ contracture found after 2 days in organ culture suggests that NE is not the only trophic influence acting on the portal vein . In addition, incubation of veins in a NE-containing medium produced a marked subsensitivity to the contractile effects of NE, but not BA2+, and thus possible desensitization of noradrenergic receptors . The data thus support a trophic role for NE in the rat portal vein.

Br J Nutr, 1980 Nov, 44(3), 371 - 80
Preparation of an experimental low-fluoride diet from single-cell organisms for rats and mice; Khalawan SA et al.; 1 . A method for producing a standard low-fluoride diet from a green alga and yeast is described . Chlorella pyrenoidosa was grown in a culture medium prepared with distilled water and analytical grade chemical salts . The spent culture medium from the alga culture was reclaimed and replenished with salts and sucrose for the production of yeast, Saccharomyces cerevisiae . 2 . The single-cell organisms were separated by centrifugation from their culture media and the dried cells were blended with sucrose, maize oil, cellulose and a salt mix to produce diet pellets for rats and mice . 3 . The diet was readily accepted as food by rats and mice and it was found to contain 100-300 micrograms fluoride/kg dry weight . Two generations of rats and four generations of mice were bred on this diet . 4 . The use of hydroxyapatite to reduce the fluoride content of the chemical used in the production of the alga and yeast biomass was investigated . Diet pellets prepared with this biomass contained 45-60 micrograms fluoride/kg dry weight.

J Clin Endocrinol Metab, 1980 Nov, 51(5), 978 - 87
Ectopic production of somatostatin-like immuno- and bioactivity by cultured human pulmonary small cell carcinoma; Szabo M et al.; Eleven continuous cultures of human pulmonary small cell carcinoma cells were examined, and eight were shown to secrete quantities of somatostatin-like immunoreactivity (SRIF-LI) ranging from 0.07-27 ng/ml culture medium/4 days, SRIF-LI was also found in a 2-N acetic acid extract of one of three human pulmonary small cell carcinomas obtained at autopsy as well as in the extract of a solid tumor resulting from inoculation of nude, athymic mice with SRIF-LI-producing, cultured small cell carcinoma cells . The SRIF-LI produced by one continuous cell line, DMS 53, was characterized in terms of its immunological, chromatographic, and biological properties . SRIF-LI from DMS 53 culture media and lysed cells was heat stable and exhibited parallel displacement to synthetic SRIF standard in a double antibody RIA . DMS 53 SRIF-LI was quantitatively retained on an immunoaffinity column of sheep anti-SRIF-Sepharose 4B under neutral conditions and could be eluted with 2 N acetic acid . Gel filtration chromatography of immunoaffinity-purified SRIF-LI revealed multiple molecular weight forms, the largest of which had an apparent molecular weight of 10,000-12,000 daltons and may represent a precursor form . This high molecular weight SRIF-LI form was resistant to exposure to denaturing conditions (8 M urea or 4 M urea plus 0.5% mercaptoethanol), suggesting the absence of noncovalent and/or disulfide linkages . A low molecular weight form coeluted with synthetic SRIF . Additional evidence for the identity of this form with the tetradecapeptide was provided by highly specific reverse phase high performance liquid chromatography . The rate of degradation of high molecular weight SRIF-LI by the cultures was markedly reduced in comparison to that of the SRIF monomer, resulting in a preferential accumulation of high molecular weight SRIF-LI in 4-day culture medium . Bioactivity of DMS 53 SRIF-LI was assessed in 4-day primary monolayer cultures of rat adenohypophyseal cells where 10(-10)-10(-9) M synthetic SRIF elicited a linear log-dose suppression of 5 X 10(-4) M synthetic SRIF elicited a linear log-dose suppression of 5 X 10(-4) M dibutyryl cAMP-stimulated rat GH release . Immunoaffinity-purified SRIF-LI from DMS 53 lysed cells and 1-h serum-free incubation medium, which consisted predominantly of monomeric SRIF, was equipotent to synthetic SRIF, SRIF-LI from 4-day culture medium consisted mostly of the high molecular weight form and exhibited a reduced bioassay potency ratio relative to synthetic SRIF of 0.73 (95% confidence limits, 0.99-0.53) . Chromatographically purified high molecular weight SRIF-LI had significant bioactivity with a bioassay to immunoassay ratio of 0.19 (95% confidence limits, 0.33-0.09) . The demonstration of ectopic SRIF, production by human pulmonary small cell carcinoma is consistent with the proposed derivation of this tumor from a cell type in the amine precursor uptake and decarboxylation cell series.

Lipids, 1980 Oct, 15(10), 838 - 48
Selectivity in incorporation, utilization and retention of oleic and linoleic acids by human skin fibroblasts; Rosenthal MD; Fetal human fibroblasts were grown in culture medium containing 10% fetal bovine serum supplemented with {1(-14)C}linoleate or {1(-14)C}oleate . At all concentrations of exogenous fatty acids, the incorporation of oleate was greater than that of linoleate . With increased medium fatty acid concentrations, linoleate in triacyglycerol (TAG) could be increased from 13 to 75% of the total incorporated; at each concentration, relatively more linoleate than oleate was in TAG . When the cells were exposed to exogenous oleate/linoleate mixtures, the composition of the mixture determined the extent of incorporation of both fatty acids . When the mixture was primarily linoleate, scarce oleate was used preferentially for phospholipids (PL); no such specificity for scarce linoleate was observed . Addition of exogenous fatty acids resulted in a shift of previously incorporated 14C fatty acids from phospholipid into TAG; retention of oleate in PL was greater than that of linoleate . Incorporation of oleate into phospholipids was also higher than that of linoleate from exogenous fatty acid mixtures which were 80% saturated . It is suggested that normal human fibroblasts have adapted to the low levels of exogenous polyunsaturated fatty acids in culture media by increased use of oleate in phospholipid . Even when the cells are supplemented with linoleate, the preferential use of oleate in phospholipid groups is retained.

J Exp Med, 1980 Oct 1, 152(4), 1070 - 84
Dendritic cells are accessory cells for the development of anti-trinitrophenyl cytotoxic T lymphocytes; Nussenzweig MC et al.; This study establishes that dendritic cells (DC) are the critical accessory cells for the development of anti-trinitrophenol (TNP) cytotoxic T lymphocytes (CTL) in vitro . We developed a model in which nylon wool-nonadherent spleen cells were used both as the responding and stimulating cells, the latter having been TNP-modified and x-irradiated . Thy-1-bearing CTL developed in C57BL/6, B6D2F1, and CBA mice only when small numbers of DC were added . Maximal responses in 5-d cultures were achieved with 0.5-1 DC/100 responding T cells . The DC did not have to be TNP modified directly . Anti-Ia and complement inactivated accessory cells, whereas similar treatment of the responders had no effect . DC exposed to ultraviolet radiation were ineffective, but x-irradiated DC were fully active . Culture media from DC, or from DC-nylon wool-passed spleen T cell cocultures that contained abundant CTL, would not substitute for viable DC . Enriched preparations of macrophages (M phi) were obtained from blood, peritoneal cavity, and spleens of BCG-immune and unprimed mice . M phi added at doses of 0.2-4% were weak or inactive as accessory cells . The level of Ia antigens on test M phi populations was quantitated and visualized by binding of a radioiodinated monoclonal anti-I-Ab,d antibody, clone B-21 . M phi that bore substantial amounts of Ia from all organs were weak accessory cells . Addition of M phi to DC-T cell cocultures produced inhibitory effects, usually at a dose of 2% M phi . In contrast, 0.5% Ia-bearing M phi from BCG-immune boosted mice inhibited > 80% of the DC-mediated CTL response . Addition of indomethacin reversed M phi inhibition, and 10(-9) M prostaglandin E2 in turn blocked the indomethacin effect . Indomethacin also restored a low level of accessory cell function in immune-boosted adherent peritoneal cells, but not in preparations of monocytes and spleen M phi . Small numbers of DC were identified in preparations of immune-boosted peritoneal cells and may have accounted for the observed accessory activity . We conclude that the development of anti-TNP CTL is an immune response in which (a) DC are the critical accessory cells; (b) Ia-bearing M phi are weak or inactive; and (c) M phi can inhibit DC-mediated response by an indomethacin-sensitive mechanism.

Surg Neurol, 1980 Oct, 14(4), 303 - 9
In vitro secretion of follicle-stimulating hormone by pituitary chromophobe adenomas; Takeuchi J et al.; Of 20 female and 14 male patients with chromophobe adenomas, 4 male patients, aged 38 to 47 years, and one female patient, aged 54, showed above normal basal plasma levels of follicle-stimulating hormone (FSH) . An in vitro tissue culture study that was performed using chromophobe cells obtained from 3 of the 4 male patients and the 1 female patient showed moderate FSH secretion into the culture media . The secretion of FSH by pituitary chromophobe adenomas might be more frequent than hitherto suspected.

Invest Ophthalmol Vis Sci, 1980 Oct, 19(10), 1204 - 21
Evidence for a role of the plasminogen activator--plasmin system in corneal ulceration; Berman M et al.; Plasminogen is present in the cornea andcan be activated to plasmin by plasminogen activator . Plasmin is able, in turn, to activate latent collagenase . This system could initiate and perpetuate the collagen degradation of corneal ulceration . This report details evidence for such a system in the cornea . Plasmin has been found to activate latent collagenase from organ cultures of ulcerating rabbit corneas and from fibroblast cultures derived from such corneas . As in the case of activation by trypsin, activation by plasmin results in the conversion of the 40,000 MW latent form to an active species of 23,000 MW . Explants of normal or alkali-burned, ulcerating corneas demonstrated plasminogen-dependent lysis of fibrin clots; frozen sections of such corneas demonstrated that lysis begins in the superficial stroma near the periphery of the cornea . Multiply freeze-thawed ulcerating corneas, but not normal corneas, showed initial lysis, not peripherally but at the ulcer region containing polymorphonuclear leukocytes . The fact that the peripheral lytic pattern existed in corneas that were obtained from eyes prefrozen in liquid nitrogen before excision of the corneas would suggest that plasminogen activator is normally contained in cells in vivo and is not made only in response to tissue injury . There was no correlation between the location of blood vessels or the presence of the corneal endothelium and the plasminogen-dependent lysis . Plasminogen activator from the ulcerating cornea and from fibroblasts was characterized by sodium dodecyl sulfate--gel electrophoresis of its cleavage products of plasminogen . The activator cleaves plasminogen into heavy- and light-chain fragments similar to those produced from plasminogen by urokinase . Plasminogen activator activity was quantitated by a new assay that restricts diffusion of the enzyme to one dimension into a narrow bore tube . The addition of plasminogen daily to cultures of ulcerating corneas resulted in earlier rises of plasminogen activator, collagenase, and collagen degradation fragments in the culture media . Although total plasminogen activator levels were not increased by the addition of plasminogen to culture, levels of both collagenase and solubilized collagen were approximately doubled . It is concluded that the plasminogen activator--plasmin system might play an important role in the destruction of stromal matrix in corneal ulceration.

J Anim Sci, 1980 Sep, 51(3), 660 - 7
Influence of culture media on in vitro fertilization of ovine tubal oocytes; Bondioli KR et al.; The ability of Synthetic Oviductal Fluid (SOF) and Minimum Essential Medium (MEM) to support sperm binding, sperm penetration and subsequent cell division of ovine tubal oocytes in vitro was evaluated . These media contained no protein, 3% (w/w) bovine serum albumin (BSA) or 20% (v/v) lamb serum (LS) . Midventral laparotomies were performed on 49 ewes synchronized with progesterone and then superovulated, and tubal oocytes were collected by flushing the oviducts . Oocytes were pooled and randomly added to 2 X 10(7) ejaculated spermatozoa in 1 ml of treatment media that had been preincubated in tubes for 3 hr at 37 C under 5% CO2 in air . After an additional 3 hr of culture in a rotating tissue culture drum, oocytes were further cultured in microdrops of the same medium for 48 hr and observed for sperm penetration, cell division and fragmentation . Oocytes were then stained with aceto-orcein and observed for penetration, multiple nuclei and bound sperm . Percentage of oocytes showing penetration in SOF, SOF+ BSA, SOF+LS, MEM, MEM+BSA and MEM+LS was 14, 3.6, 7.8, 11.1, 8.7 and 12.8, respectively; percentage of oocytes cleaving was 8, 3.6, 5.9, 4.4, 4.3 and 4.3; percentage of oocytes fragmenting was 40, 60, 60.8, 20, 60.9 and 29.8, and the average number of sperm cells bound per oocyte was 82, 177, 117, 41, 56 and 47 . No cell division was observed for control oocytes (no sperm added), but 72.7, 80 and 76.9% fragmented in SOF, SOF+BSA and SOF+LS, respectively . No difference was found among the media in their ability to support sperm penetration and subsequent cell division in vitro . All media tested supported a high incidence of sperm binding and oocyte fragmentation.

Atherosclerosis, 1980 Sep, 37(1), 11 - 9
Enhanced synthesis of collagen and total protein by smooth muscle cells from atherosclerotic rabbit aortas in culture; Pietila K et al.; Protein synthesis in smooth muscle cells from normal and atherosclerotic rabbit aortas was studied . Cultures of cells from the third passage were incubated with {3H}proline and the synthesis rates of collagen and total protein measured from the radioactivities incorporated into protein-bound hydroxyproline and proline, respectively . The synthesis ratio of collagen types I and III was studied by separating the radioactive procollagens of the culture media by DEAE column chromatography . The synthesis rates of both collagen and total protein were higher in cells cultured from atherosclerotic rabbit aortas . There was no difference in the synthesis ratios of types I and III collagen between cells from the two origins . In addition to procollagens, a third protein eluted before procollagen I in the DEAE chromatography of medium proteins . Compared with control cells, cells from atherosclerotic aortas incorporated relatively less radioactivity into this protein . No differences were detected in the amino acid compositions of cellular proteins between cells from both origins . It was concluded that cells grown from atherosclerotic rabbit aortas synthesize more collagen and total protein than those from normal aortas, the synthesis of collagens I and III being equally enhanced . The results also suggest that there are proteins which are not involved in the general enhancement of protein synthesis.

Endocrinology, 1980 Sep, 107(3), 839 - 44
Glucocorticoid dependence of fetal lung maturation in vitro; Torday JS; Earlier studies of the fetal lung in tissue culture have led to the conclusion that its maturation is autonomous . We have studied the role of the small amount of cortisol supplied by the serum routinely added to culture media . It was observed that 28-day-old fetal rabbit lung cells continued to grow and differentiate in culture medium supplemented with 10% calf serum (2.2 X 10(-8) M cortisol) . However, when these cells were propagated in medium supplemented with calf serum which was charcoal stripped or from adrenalectomized animals, growth and differentiation were prevented (<1 X 10(-12) M cortisol) . The addition of cortixol (1 X 10(-10) to 1 X 10(-5) M) to such media restored the ability of these cells to grow and differentiate in culture . Other classes of steroid hormones failed to restore such activity even with a 10-fold excess of cortisol . We conclude that maturation of the 28-day-old fetal rabbit lung in vitro is dependent upon the presence of cortisol.

J Cell Physiol, 1980 Sep, 104(3), 349 - 57
Regulation of prostaglandin synthesis during differentiation of cultured mouse myeloid leukemia cells; Honma Y et al.; Mouse myeloid leukemia cells (Ml) were induced to differentiate into mature macrophages and granulocytes by various inducers . The differentiated Ml cells synthesized and released prosetaglandins, whereas untreated Ml cells did not . When the cells wee prelabelled with {14C}arachidonate, the major prostaglandins released into the culture media were found to be prostaglandin E2, D2, and F2 alpha in an early stage of differentiation, but the mature cells produced predominantly prostaglandin E2 . The synthesis and release of prostaglandins were completely inhibited by indomethacin . Dexamethasone, a potent inducer of differentiation of Ml cells, did not induce production of prostaglandins in resistant Ml cells that could not differentiate even with a high concentration of dexamethasone . These results suggest that production of prostaglandins in Ml cells is closely associated with differentiation of the cells . Homogenates of dexamethasone-treated Ml cells converted arachidonate to prostaglandins, but this conversion was scarcely observed with homogenates of untreated Ml cells . Dexamethasone and the other inducers stimulated the release of arachidonate from phospholipids . Therefore, induction of prostaglandin synthesis during differentiation of Ml cells may result from induction of prostaglandin synthetase activity and stimulation of the release of arachidonate from cellular lipids . Lysozyme activity, which is a typical biochemical marker of macrophages, was induced in Ml cells by prostaglandin E2 or D2 alone, as well as by inducers of differentiation of the cells, but it was not induced by arachidonate or prostaglandin F2 alpha . These results suggest that prostaglandin synthesis is important in differentiation of myeloid leukemia cells.

J Biol Chem, 1980 Aug 10, 255(15), 7071 - 4
NH2-terminal sequence analysis of pro-C4, the precursor of the fourth component of guinea pig complement; Goldberger G et al.; The amino acid sequence of the NH2-terminal regions of the intracellular precursor of the fourth component of guinea pig complement (pro-C4) and isolated alpha, beta, and gamma chains of native C4, synthesized by peritoneal macrophages in culture, were determined with a microradiosequencing technique . Radiolabeled pro-C4 was immunoprecipitated from cell lysates and native C4 from culture media which contained one of six 3H-amino-acids or {35S}methionine . The purity of these proteins was established and molecular weights were estimated by electrophoresis in sodium dodecyl sulfate . Seventeen of the first 22 residues of pro-C4 were identified . Fifteen of these 17 were nonpolar . The eight amino acids ascertained within the first 9 NH2-terminal residues of guinea pig pro-C4 were identical with those reported for human C4 beta chain, and identity with residues determined for guinea pig C4 beta chain was established . These data indicate that beta chain is the NH2-terminal segment of pro-C4, that no residues are cleaved from the NH2 terminus during the conversion to native C4, and that this segment of the pro-C4 molecule is conserved phylogenetically.

Science, 1980 Aug 8, 209(4457), 701 - 2
Estrogen and the growth of breast cancer: new evidence suggests indirect action; Shafie SM; The growth of the MCF-7 human breast cancer cell line is unresponsive to the presence of estrogen in culture media . Paradoxically, in nude mice, growth of these cells and formation of solid tumors are dependent on estrogen . Tumors fail to develop in ovariectomized mice, but do develop in intact mice and in ovariectomized mice given estrogen . Primary cultures derived from MCF-7 tumors revert to unresponsiveness to estrogen . However, when these cultures are again transplanted into nude mice, estrogen is required for tumor formation . The continuous culture, the solid tumor, and the primary cultures therefrom have similar estrogen-binding capacities and affinities . These results indicate that mammary carcinoma cell growth in vivo is subject to inhibition that can be overcome by estrogen.

In Vitro, 1980 Aug, 16(8), 669 - 74
An improved culture medium for mouse blastocysts; Spindle A; Eagle's basal medium, modified to contain essential amino acids at the concentrations optimal for mouse blastocyst hatching, attachment, and outgrowth, supported in vitro development of the mouse blastocyst better than other tissue culture media tested . This medium was improved for growth and differentiation of the inner cell mass by doubling the concentration of amino acids and glucose and by adding uridine (10(-5) M) and beta-mercaptoethanol (10(-5) M) . In this improved medium nearly all blastocysts grown from the two-cell stage hatched and formed trophoblast outgrowths, and 62% developed into two-layer egg cylinders.

Appl Environ Microbiol, 1980 Aug, 40(2), 391 - 9
Cholesterol as a limiting factor in the growth of Mycoplasma pneumoniae; Johnson JK et al.; Ultracentrifugation was used to separate three commercial lots of bovine serum fraction (BSF) into components designed to contain lipoproteins . Each BSF lot and component was tested for ability to support the growth of tree strains of Mycoplasma pneumoniae . In general, the level of growth-promoting activity corresponded to the amount of cholesterol present in the BSF or BSF components rather than to the amount or type of lipoprotein Cholesterol was the limiting nutritional factor of BSF with low growth-promoting activity . The addition of cholesterol and bovine serum albumin to BSF with low activity resulted in growth equal to or greater than that observed for BSF with high growth-promoting activity . When cholesterol was added to agar medium containing BSF of low activity, mycoplasma colonies were greater in number, possessed larger mean diameters, and had centers that were more distinct than those observed when this BSF was used alone . Variability in growth-promoting actions of commercial lots of BSF was eliminated by increasing their cholesterol content to an optimum level . An adjustment of the cholesterol and albumin levels of any serum product used in culture media may provide a simple convenient method to improve growth and isolation of mycoplasmas.

Am J Anat, 1980 Aug, 158(4), 433 - 444
Maintenance of gonadotrophs in pituitary autografts under the kidney capsules of female rats given sex hormones or LRH; Shino M et al.; Female Sprague-Dawley rats were hypophysectomized and the anterior pituitary gland was immediately placed under the kidney capsule . For 1 week after surgery, groups of pituitary autograft-bearing animals were treated with twice-daily injections of estradiol 17 beta (E), progesterone (P), estradiol 17 beta and progesterone (EP), or luteinizing hormone-releasing hormone (LRH) . Within 2--4 hours following the last injection, the pituitary grafts were removed and placed into organ culture . They were maintained in culture with or without added LRH (10(-7) M) for 1 hour at 37 degrees C . The culture media were then frozen for later radioimmunoassay of FSH and LH . The tissues were kept in culture for an additional 24 hours, at which time they were fixed and prepared for immunocytochemistry or electron microscopy . Results showed that treatment of the animals with E, EP, or LRH enhanced the release of FSH and LH into the culture media, and that the release of these hormones was increased further by acute incubation with LRH . The ultrastructure of the gonadotrophs was well maintained by treating the animals with E or the combination of E and P or with LRH . Graft tissue from animals treated with LRH, which was incubated subsequently for 24 hours with LRH, showed the best maintenance of gonadotroph morphology . This experimental procedure should be useful for obtaining gonadotrophs for use in establishing gonadotroph cell lines.

J Biol Chem, 1980 Jul 25, 255(14), 6579 - 83
Secretion of the vitamin K-dependent protein of bone by rat osteosarcoma cells . Evidence for an intracellular precursor; Nishimoto SK et al.; Four clonal cell lines derived from a rat osteosarcoma were tested for the ability to secrete the gamma-carboxyglutamic acid-containing protein of bone (BGP) using a specific radioimmunoassay for this protein . Two cell lines secreted BGP into culture media while the other two did not . Other investigators have shown that these two cell lines are also the only ones with the high parathyroid hormone responsiveness and alkaline phosphatase activity expected for osteoblast cells in culture . Both cell lines also form a mineralized sarcoma when implanted in rats . The BGP in culture media is identical in molecular weight and in electrophoretic mobility with the 5800-dalton BGP purified from rat bone . Thus, BGP is probably secreted by osteosarcoma cells directly and not derived from an extracellular precursor by proteolytic cleavage . There are two immunoreactive components within osteosarcoma cells which secrete BGP . One component is identical in molecular weight and electrophoretic mobility with BGP from rat bone . The other component has a higher molecular mass (approximately 9000 daltons) and about half the electrophoretic mobility of BGP from bone . The presence of both components within these cells raises the possibility that the larger component may be an intracellular precursor which is processed to BGP prior to secretion.

J Biol Chem, 1980 Jul 10, 255(13), 6036 - 9
Undersulfated proteoglycans are secreted by cultured chondrocytes in the presence of the ionophore monensin; Tajiri K et al.; The monovalent ionophore, monensin, inhibits secretion of many different proteins from a wide variety of cells . The site of blockage is at the golgi complex . We have exposed chick embryo chondrocytes in suspension culture to monensin, at concentrations ranging from 10(-8) to 10(-6) M . At the higher concentrations, between 10(-7) and 10(-6) M, monensin inhibited secretion of type II procollagen, which accumulated in the chondrocytes . At these concentrations of the ionophore, proteoglycan synthesis was inhibited, as measured by radioactive serine incorporation into core proteins and by radioactive glucosamine or SO4 incorporation into glycosaminoglycans . However, at a monensin concentration of 3 x 10(-8) M, the incorporations of serine and glucosamine were close to normal while SO4 incorporation was at 30% of control values . The ratio of glucosamine to serine in pronase-released glycosaminoglycans from culture media was unaffected by 3 x 10(-8) M monensin but the sulfate to serine ratio decreased to 29% of control values . Examination of the glycosaminoglycans by gel filtration showed a progressive increase in Kav values as sulfation decreased . Undersulfation was demonstrated by radiochromatographic analysis of the digestion products following incubation with chondroitinase ABC . The composite results show that monensin interferes with sulfation of newly synthesized proteoglycans.

J Cell Physiol, 1980 Jul, 104(1), 41 - 6
Murine sarcoma growth factors affect the growth and morphology of cultured mouse epithelial cells; Keski-Oja J et al.; A mouse embryo epithelial cell line, MMC-E, was used to study the effects of purified murine sarcoma growth factors (SGFs) on the growth of epithelial cells . Murine SGF was a partially purified preparation from the serum-free culture media of mouse fibroblasts transformed by Moloney murine sarcoma virus . SGFs stimulated DNA-synthesis in resting cells and induced them to grow to higher densities than control cells . With continued exposure to SGFs, MMC-E cells lost their postconfluency inhibition of division and formed small expanding foci . When the SGFs were removed and the cells were subcultured, they regained their normal phenotype, showing that the effects of SGFs are reversible on these epithelial cells . SGFs could also stimulate the MMC-E epithelial cells to grow in soft agar, like the syngeneic fibroblasts (MMC-F) from the same mouse embryo, but slower than the control fibroblastic clone . Microgram quantities of SGFs were needed to stimulate soft agar growth of MMC-E cells . The results indicate that SGF can bring about a phenotypic change in the growth pattern of epithelial cells as well as fibroblastic cells.

Biomedicine, 1980 Jul-Aug, 33(4), 103 - 5
Effect of levamisole on granulopoiesis in agar culture; Hellmann A et al.; We studied the effect of levamisole on the proliferation of granulocyte-committed progenitor cells (CFU-c) in the agar system . Media conditioned by pooled human peripheral blood mononuclear cells served as source of colony-stimulating activity (CSA) . Levamisole at concentrations ranging from 0.1 to 10.0 micrograms/ml had no detectable stimulatory or inhibitory effect on granulopoiesis in vitro . Levamisole added to culture media neither enhanced nor reduced the release of mononuclear cells of CSA . We conclude that the cases of granulocytopenia, occasionally severe, associated with levamisole therapy are more likely to be due to individual idiosyncrasy or hypersensitivity then to dose-related myelotoxicity.

J Clin Microbiol, 1980 Jul, 12(1), 74 - 8
Monoclonal antibodies specific for dengue virus type 3; Dittmar D et al.; Mouse lymphocyte hybridomas were prepared by polyethylene glycol-mediated fusion of cells from a mouse plasmacytoma line with lymphocytes from a mouse hyperimmunized with dengue virus type 3 (dengue-3) . Media from 50 hybrid colonies were screened; 46 of them showed antibody activity against dengue-3-infected cells as determined by an indirect immunofluorescent antibody technique . Dengue monoclonal antibody obtained after cloning one of these colonies demonstrated activity in hemagglutination inhibition and indirect immunofluorescent antibody assays with dengue-3 antigen, but not type 1, 2, and 4 antigens . In addition, this antibody activity could be removed from culture media only by absorption with dengue-3 antigen.

Acta Physiol Scand, 1980 Jul, 109(3), 275 - 81
Effects of purified macrophage RNase on granuloma fibroblasts with reference to silicosis; Aho S et al.; Two alkaline RNases, designated RNase 1 and RNase 2, were isolated from the culture media of silica-treated and non-treated macrophages . The yield of RNase from the medium of silica-treated macrophages was 30% of that from the non-treated control . The effects of these RNases on cultured granuloma fibroblasts and on granulation-tissue nuclei were studied . RNase 1 inhibited thymidine incorporation into fibroblasts except at low concentrations, where it was observed to be stimulatory . RNase 1 also inhibited the protein synthesis of fibroblasts . The incorporation of cytidine into RNA in cultured fibroblasts was not affected by RNase 1, but the incorporation into isolated nuclei was decreased . In pulse chase experiments RNase 1 increased the release of cytidine, but not that of thymidine, from the cells . RNase 2 had no effect on the protein or nucleic acid metabolism of the fibroblasts or on the RNA metabolism of isolated nuclei, perhaps because of impermeability . These experiments confirm that macrophage RNase activity is able to regulate the metabolism of granulation-tissue fibroblasts by increasing RNA degradation . Through this action it also regulates DNA and protein synthesis and other metabolic functions of those cells.

J Gen Virol, 1980 Jul, 49(1), 83 - 9
Enzyme-linked immunosorbent assay for coronaviruses HCV 229E and MHV 3; Kraaijeveld CA et al.; The antigenic relationship between human cornonavirus strain 229E (HCV 229E) and mouse hepatitis virus strain 3 (MHV 3) was studied by means of the indirect form of enzyme-linked immunosorbent assay (ELISA) . A cross-reaction was found with hyprimmune rabbit sera between HCV 229E and MHV 3 which may be due to the adherence of bovine serum componeants from tissue culture media, which were present on virus particles even after extensive purification . No cross-reaction was observed with immune sera absorbed with bovine serum, or with HCV 229E grown in tissue culture without serum . This indirect ELISA with HCV 229E may prove to be useful for studies with human sera.

J Clin Endocrinol Metab, 1980 Jul, 51(1), 182 - 4
Evidence for a peptide similar to 16K fragment in man . Its relationship to ACTH; Bertagna X et al.; A radioimmunoassay directed toward the NH2-terminal region of mouse pro-ACTH/endorphin (called 16K fragment) was used to examine human samples . Culture media from two corticotropic adenomas and plasmas from 11 patients with various ACTH hypersecretory syndromes gave parallel displacement curves; displacement curves for human samples were not parallel to purified mouse 16K fragment . Following sodium dodecyl sulfate polyacrylamide gel electrophoresis of culture medium from one adenoma, a major peak of 16K fragment immunoreactivity with an apparent molecular weight of ca . 16,000 was detected . A significant correlation (r = 0.963 ; p less than 0.001) was found between immunoreactive 16K fragment and ACTH in the patients' plasmas . These data indicate that a peptide similar to 16K fragment exists in man ; that human and mouse 16K fragment are immunologically distinguishable and that human 16K fragment appears to be secreted concomitantly with ACTH.

J Biol Chem, 1980 Jun 25, 255(12), 5681 - 7
Human adenosine deaminase and its binding protein in normal and adenosine deaminase-deficient fibroblast cell strains; Daddona PE et al.; Reduced or absent adenosine deaminase activity occurs in all tissues of some patients with severe combined immunodeficiency disease . In this report we describe (a) a sensitive and specific radioimmunoassay for fibroblast adenosine deaminase and its binding protein, (b) the importance of various tissue culture media on the expression of fibroblast adenosine deaminase, and (c) the levels of immunoreactive adenosine deaminase compared to its binding protein in normal and homozygous adenosine deaminase-deficient fibroblast cell strains.

Diabetes, 1980 Jun, 29(6), 497 - 500
Monolayer cultures of adult pancreatic islet cells on osmotically disrupted fibroblasts; Meda P et al.; A simple method that permits the attachment of fully dissociated adult pancreatic islet cells to culture dishes, using osmotically disrupted fibroblasts, is described . Using this system, monolayer cultures consisting of either isolated or clustered adult endocrine islet cells can be readily established in standard culture media.

Scand J Clin Lab Invest, 1980 Jun, 40(4), 311 - 8
Liberation of a fibrogenic factor from human blood monocytes, ascites cells, cultured histiocytes and transformed mouse macrophages by treatment with SiO2; Aalto M et al.; Human monocytes and ascites macrophages from cirrhotic patients were isolated in Percoll-gradient and cultured with and without silica . Similar experiments were carried out also with cultured malignant human histiocytes and transformed mouse macrophages . The fibrogenic activity of the culture media was tested by measuring the incorporation of {3H}proline and {3H}thymidine into cultured rat granuloma and human synovial cells . Media from silica-treated monocytes, ascites macrophages and certain histiocyte and mouse macrophage lines caused an increase in the incorporation of both {3H}proline and {3H}thymidine into collagen and DNA, respectively, in both cell systems . Alkaline RNase activities were decreased markedly in the media from silica-treated ascites macrophages but not in the media of the monocytes or histiocytes.

Biochim Biophys Acta, 1980 May 28, 618(2), 347 - 58
Production of apolipoproteins E and A-I by rat hepatocytes in primary culture; Dashti N et al.; Hepatocytes were isolated from normal adult rat livers and cultured in a modified HI-WO/BA medium . A nearly confluent monolayer was established at the plating concentration employed . The hepatocytes synthesized ansd secreted albumin at rates similar to those observed in vivo . The cells secreted triacylglycerol in the absence of fatty acid substrate . Under these conditions the most abundant triacylglycerol molecular species contained 53 carbons . Incubation with oleic acid markedly increased triacylglycerol secretion predominantly in the form containing a total carbon number of 57 . Approx . 80% of the secreted cholesterol was in the free form and this was unaffected by oleic acid . Employing monospecific antibodies constant rates of synthesis and secretion of apolipoproteins E and A-I were demonstrated by quantitative electroimmunoassay of the cell culture media . The rates of albumin, apolipoprotein E, and apolipoprotein A-I production were 1480, 170 and 60 microgram/h per g cell protein, respectively.

Gastroenterology, 1980 May, 78(5 Pt 1), 925 - 30
Effects of carbachol and atropine on gastrin secretion and synthesis in rat antral organ culture; Harty RF et al.; The purpose of the present study was to examine the effects of the cholinergic agent carbachol on gastrin synthesis and secretion by rat antral mucosa in organ culture . In addition, the effect of atropine on basal and carbachol-stimulated gastrin release was investigated . These in vitro studies demonstrate that carbachol stimulated both gastrin secretion and synthesis in a dose-dependent manner . Maximal stimulation of gastrin synthesis and release occurred at a concentration of 1 X 10(-5) M carbachol . The rate of gastrin release stimulated by carbachol (1 X10(-5) M) was 0.79 ng-hr-1-mg-1 which was significantly greater than the control value of 0.37 ng-hr-1 (P less than 0.001) . Atropine, over the dose range from 10(-7) to 10(-3) M, had no effect on basal (unstimulated) gastrin release . However, carbachol-stimulated gastrin release was inhibited progressively by inclusion of increasing concentrations of atropine in the culture media . The results of these studies indicated that the acetylcholine analogue, carbachol, is capable of directly stimulating the antral gastrin cell to significantly increase the rate of synthesis and release of gastrin . Whereas atropine, under these in vitro conditions, does not alter basal gastrin release, the muscarinic antagonist does competively inhibit carbachol-stimulated gastrin secretion.

J Pediatr, 1980 May, 96(5), 837 - 41
X-linked mental retardation, macro-orchidism, and the Xq27 fragile site; Turner G et al.; Twenty-three families with X-linked mental retardation were examined for the presence of a fragil site on the long arm of the X chromosome (Xq27 fra) . Specific culture media were necessary to demonstrate this site . In only seven of the families was the Xq fragile site observed; in these, all of the affected males had both the fragile X and macro-orchidism . Macro-orchidism was not observed in the remaining 16 families . In the families with Xq27 fra segregating the fraes . This correlated with the age of the carrier . The 25 affected males with macro-orchidism and Xq27 fra had some minor clinical features in common: there was an increase in birth weight, high forehead, prognathism, pale irides, big ears, and an increased head circumference in infancy and childhood which did not persist into adult life . The majority of the affected individuals were moderately retarded.

Biomedicine, 1980 May, 33(3), 61 - 2
Effect of antiobiotics on elimination of mycoplasma from experimentally infected cells; Negre G et al.; The primary and continuous cell cultures and mouse peritoneal macrophages were experimentally infected by Mycoplasma pneumoniae and M . orale I strains and maintained in culture media containing high concentrations of tetracycline or kanamycin . The viable mycoplasmas were eliminated from cell supernatants but not from cells.

Ann Endocrinol (Paris), 1980 May-Jun, 41(3), 157 - 92
{Recent data on somatomedins (insulin-like growth factors) (author's transl)}; Binoux M; Data on somatomedins have resulted from the convergence of two initially distinct lines of research relating to the two major aspects of their biological activities, their effects on cartilage growth and their insulin-like action . Human serum has yielded three factors, SM-A, SM-C and NSILA-S . The latter comprises two very similar polypeptides, IGF I and IGF II which structurally closely resemble proinsulin . A fourth factor, MSA, has been isolated from calf serum and from conditioned medium of a rat liver cell strain . All these chemically and biologically closely related factors have a molecular weight of approximately 7 500 but circulate in a form with much higher molecular weight owing to their binding to specific carrier proteins . Over the past five years, the use of purified somatomedins has led to rapid progress in the elucidation of the physiology of these hormones . In this review, present knowledge of somatomedins is analysed in the following terms: physiochemical properties, circulating forms, action on growth, insulin-like activity, interaction with receptors, immunological characteristics, biosynthesis and hormonal regulation of their blood level . Results obtained in our laboratory from rat liver organ culture are discussed: the action of culture media on cartilage sulphation, the detection of a protein which specifically binds NSILA-S and SM-A and its use in a competitive protein binding assay for somatomedins in biological fluids.

Arch Ophthalmol, 1980 May, 98(5), 919 - 23
Collagenase levels in a new model of experimental herpetic interstitial keratitis; Campbell R et al.; A reproducible model of severe herpetic interstitial keratitis was developed by injecting live herpes simplex virus type 2 intrastromally into the corneas of presensitized rabbits . Herpes-infected corneas showed significantly more stromal infiltration and vascularization, iritis, conjunctivitis, and epithelial disease than the control corneas injected with cell supernatant without virus . Levels of total collagenase detected in the culture media of the herpes-infected corneas were high and were similar to those observed previously in alkali-burned rabbit corneas . Unlike alkali-burned corneas, the herpes-infected corneas showed much more of the enzyme in the latent form during the first two days of culture.

J Cell Physiol, 1980 Apr, 103(1), 47 - 54
Fibronectin from aged fibroblasts is defective in promoting cellular adhesion; Chandrasekhar S et al.; Late passage fibroblasts show decreased cell-substrate adhesion . We provide evidence that the reduced adhesion is due to a defect in the adhesive glycoprotein fibronectin . Late passage cells become more adhesive in culture media that has been conditioned by the growth of early passage cells . Analysis of fibronectins purifed from early and late passage cell conditioned media indicates that there are striking differences in their abilities to promote cell adhesion . Young cell fibronectin supports the maximal adhesion of both young and old cells . However, old cells require quantitatively more fibronectin . In contrast, old cell fibronectin is less effective in supporting the adhesion of either cell type . In addition, neither cell type achieves a normal morphology in the presence of old cell fibronectin . The results support the conclusion that the fibronectin released by late passage cells i defective and does not support normal cell-substrate interactions.

In Vitro, 1980 Apr, 16(4), 330 - 6
Evaluation of growth hormone production from GH1 cells in vitro: effect of culture media and time in culture; Bolton WE et al.; Growth hormone production by a rat pituitary tumor cell line (GH1) was measured during lag, exponential, and plateau phases of growth in different culture media . Growth hormone secretion was low during lag and early exponential phase; it increased late in the exponential phase and continued to increase during the plateau phase . This biphasic pattern of growth hormone production was observed in all media and sera utilized . Both the doubling time and growth hormone production were influenced by the choice of media and sera . In addition, the length of time in culture affected the growth fraction with passage level 40 GH1 cells having a 79% growth fraction, whereas the growth fraction of passage level 100 cells was 95% . Using the population doubling time as a criterion for a choice of medium, F-10 medium supplemented with 20% fetal bovine serum consistently yielded the most rapid doubling time (32 hr), whereas Dulbecco's MEM supplemented with 15% horse serum and 2.5% fetal bovine serum yielded the greatest plateau cell density . Growth hormone secretion and the population doubling times were directly related to culture conditions including length of time in culture, choice of tissue culture media, choice of sera, and the phase of cell growth (lag, exponential or plateau).

Cancer Res, 1980 Apr, 40(4), 1084 - 90
Effect of fatty acid modifications of cultured hepatoma cells on susceptibility to complement-mediated cytolysis; Yoo TJ et al.; The fatty acid composition of Morris Hepatoma 7777 cells was modified by exposure to culture media that were supplemented with 0.1 to 0.36 mM oleic or linoleic acid for 5 days . Changes occurred in the fatty acid composition of both the cellular phospholipid and the neutral lipid fractions . Exposure to linoleic acid caused a large increase in the polyunsaturated fatty acid content of the cell lipids, whereas enrichment in monoenoic fatty acids occurred when the cells were exposed to high levels of oleic acid . Cellular phospholipid content decreased, cholesterol content did not change, and triglyceride content increased as a result of fatty acid supplementation . The fatty acid-modified cells showed increased susceptibility to complement-mediated cytolysis as compared with control cells grown in unsupplemented culture media . The extent of the increase in susceptibility to cytolysis depended on the degree of lipid modification and also on the cell number and antibody titer used in the assay . Cells enriched with linoleic acid were the most susceptible, but oleic acid enrichment also produced increased susceptibility to immune cytolysis . The kinetic study showed that the initial rate of cytolysis was higher in fatty acid-modified cells than in the control cells . There was no difference in the osmotic fragility of the control and fatty acid-modified cells . These results indicate that changes in the lipid composition of a target cell can influence its susceptibility to complement-dependent cytolysis.

Appl Environ Microbiol, 1980 Apr, 39(4), 818 - 22
Inhibitory effects of spices on growth and toxin production of toxigenic fungi; Hitokoto H et al.; The inhibitory effects of 29 commercial powdered spices on the growth and toxin production of three species of toxigenic Aspergillus were observed by introducing these materials into culture media for mycotoxin production . Of the 29 samples tested, cloves, star anise seeds, and allspice completely inhibited the fungal growth, whereas most of the others inhibited only the toxin production . Eugenol extracted from cloves and thymol from thyme caused complete inhibition of the growth of both Aspergillus flavus and Aspergillus versicolor at 0.4 mg/ml or less . At a concentration of 2 mg/ml, anethol extracted from star anise seeds inhibited the growth of all the strains.

Proc Natl Acad Sci U S A, 1980 Apr, 77(4), 2084 - 7
Control of growth of estrogen-sensitive cells: role for alpha-fetoprotein; Soto AM et al.; The role played by alpha-fetoprotein (AFP) during the perinatal period in rats has not yet been ascertained despite earlier suggestions that this plasma protein affected the multiplication, in an "in animal-in culture" system of tumor cells that are stimulated by estrogen (E) for growth . To test this inference developed from our previous work, AFP was purified by reverse affinity chromatography to homogeneity by electrophoretic and immunochemical criteria . Purified AFP was added at different concentrations to horse serum-supplemented medium which by itself is able to allow maximal exponential growth of a rat pituitary tumor cloned cell line C29RAP . These cells carry estrophilins and their growth is stimulated by E in animals but not in culture . At 3 mg/ml in culture media, AFP prevented growth of C29RAP cells; the effect was dose dependent . F4C1, a rat pituitary cloned cell line that carries estrophilins but shows autonomous behavior when injected into male and female Fisher rats, was not affected in cultured by comparable concentrations of AFP in the culture media . The effect of AFP on the growth of E-sensitive cells in culture was not reversed by E administration . We conclude from these experiments that (i) AFP is a specific inhibitor of the cell multiplication of cells that are E-sensitive for growth (as defined in this presentation), (ii) estrophilins seem not to play a significant role in this inhibition, and (iii) E appears not to be a growth-promoting hormone per se.

Nippon Naibunpi Gakkai Zasshi, 1980 Mar 20, 56(3), 262 - 73
{Osmotic pressure and angiotensin II stimulation of arginine vasopressin release from a guinea pig hypothalamo-neurohypophyseal complex in organ culture (author's transl)}; Ishikawa S et al.; An organ culture system of a male guinea pig hypothalamo-neurohypophyseal complex (HNC) was established . On day 5 in culture, (Na+ -K+) ATPase activity was 0.83 +/- 0.11 mM Pi/mg prot/hr (mean +/- SEM): that is 67% of that on day 1 in culture . 3H-thymidine incorporated into DNA in the explants of HNC was 1,205 +/- 185 cpm/microgram DNA . The explants responded to the elevated KCl medium and the hypertonic solution of sodium chloride with a 470 +/- 38% and 298 +/- 31% increase in arginine vasopression (AVP) release, respectively . This response was inhibited by the addition of tetrodotoxin to the culture medium . AVP release from the explants in res-onse to angiotensin II increased significantly in a dose dependent manner . {Sar1, Ile8} angiotensin II, however, attenuated the response of the explants to angiotensin II when administered simultaneously with angiotensin II . These results suggest that angiotensin II and its analogue cause the AVP release from the explants in a competitive manner . The concentrations of AVP in the culture media made hypertonic with sodium chloride, sucrose and mannitol were 298 +/- 31% (p less than 0.01), 251 +/- 36% (p less than 0.01) and 255 +/- 59% (p less than 0.05) of their control values, respectively . The hypertonic solutions of sodium chloride, sucrose and mannitol caused AVP release from the explants in vitro, while the hypertonic solutions of glucose and urea were revealed to be poor osmotic stimuli on AVP release . These results support the concept of osmoreceptors to release AVP from the hypothalamo-neurohypophyseal axis.

Mycopathologia, 1980 Mar 17, 70(2), 125 - 8
Production of aflatoxin by Aspergillus flavus (UBMI) in some Nigerian indigenous beverages and foodstuffs; Alozie TC et al.; Sixteen samples of some Nigerian indigenous beverages and foodstuffs were analyzed for their aflatoxin content . All the eight samples of beverages tested were found to be contaminated with aflatoxin . Of the eight samples of foodstuffs tested, all contained aflatoxin except ewedu, dawadawa and shoko yokoto . All the beverages used as culture media for Aspergillus flavus (UBMI) supported the growth of the fungus and aflatoxin elaboration . A . flavus was found to grow luxuriantly on all samples of foodstuffs, except dawadawa . However, growth of the fungus on foodstuffs was not synonymous with aflatoxin production.

Biochem J, 1980 Mar 15, 186(3), 971 - 5
Effect of tunicamycin and cycloheximide on the secretion of acid hydrolases from I-cell cultured fibroblasts; Miller AL et al.; I-cell cultures fibroblasts secrete excessive amounts of N-acetyl-beta-D-hexosaminidase and alpha-L-fucosidase into the culture media as compared with normal fibroblasts . Addition of tunicamycin or cyd {14C}leucine (40--50%) into trichloroacetic acid-precipitable material decreased the secretion of these I-cell hydrolases to normal values within 24 h, but had no effect on the secretion of acid hydrolases from normal fibroblasts . These results indicate that I-cell cultured fibroblasts secrete at least two types of acid hydrolases: one is tunicamycin- and cycloheximide-sensitive and constitutes the greater proportion of the secreted hydrolases, and a smaller proportion is insensitive to tunicamycin and cycloheximide, similar t9 the acid hydrolases secreted by normal cultured fibroblasts.

Am J Vet Res, 1980 Mar, 41(3), 372 - 6
Inhibition of lymphocyte blastogenesis by sera from cows with lymphoma; Jacobs RM et al.; The sera from cows with lymphoma inhibit the blastogenesis of normal lymphocytes . The inhibitory sera were partially characterized . Twenty of 34 sera from cows with lymphoma caused inhibition, whereas 3 of 25 sera from healthy animals were inhibitory . Sera from 15 cows with various inflammatory conditions did not cause inhibition . Seventy-four cows were tested for responsiveness to phytohemagglutinin, and of this group, the stimulation indices of 25 animals which had antibody to the bovine leukemia virus did not differ from the remainder of the group . The inhibitory substance: (a) was heat stable at 56 C for 30 minutes, (b) was not overcome by the addition of phytohemagglutinin, (c) was active in a manner that was proportional to the concentration of serum in the culture media, (d) was not lymphocytotoxic at a concentration which was profoundly inhibitory to blastogenesis, (e) was found in the first peak eluted from a Sephadex G-200 column, (f) was nonspecifically active because it inhibited the blastogenesis of a panel of lymphocytes from healthy animals, (g) was present in 59% of cows with lymphoma in the terminal stages of disease, (h) was rarely present in healthy cows or cows with inflammatory conditions, and (i) was not associated with bovine leukemia virus infection in the absence of tumor development.

Acta Paediatr Scand, 1980 Mar, 69(2), 263 - 7
An arthropathic form of osteogenesis imperfecta; Penttinen R et al.; A 14-year-old girl gradually developed severe osteoporosis and destructive generalized joint disease, resulting in joint stiffness and anchyloses . She also had moderate hydroxyprolinemia and hydroxyprolinuria . Rheumatoid arthritis was highly unlikely . Anamnestic data revealed two long bone fractures . Collagen biosynthesis was studied in fibroblasts cultured from the patient's skin . Chromatograms of 3H-labelled culture media proteins on ion exchange celluloses revealed an increased ratio of type III collagen to type I collagen when compared with the chromatograms of age-matched control fibroblasts . This finding is typical of certain cell strains in osteogenesis imperfecta . The patient might thus express a new variety of osteogenesis imperfecta with chronic arthropathy.

Clin Sci (Lond), 1980 Mar, 58(3), 201 - 10
Prostaglandins as mediators of bone resorption in renal and breast tumours; Greaves M et al.; Amounts of prostaglandin E and prostaglandin F have been measured by radioimmunoassay in extracts of renal cortical carcinoma and benign and malignant breast tumours after solvent extraction and column chromatography . 2 . Substantial amounts of prostaglandin E were found in extracts of benign and malignant breast tumours and in renal tumours . Much lower amounts of prostaglandin F were present in all tumour types . 3 . Co-cultivation of tumour explants with mouse calvaria led to significant bone resorption in 10 of 13 renal carcinomas, three of eight malignant breast tumours, and two of nine benign breast tumours . Tumours associated with bone resorption had higher concentrations of prostaglandin E in culture media at the end of incubation than did non-resorbers . 4 . Indomethacin (14 mumol/1) greatly reduced bone resorption in the presence of the tumour, but this was not always complete, particularly with breast tumours . Indomethacin had no effect on prostaglandin-induced bone resorption . Theophylline (2.2 mmol/1) significantly increased prostaglandin E production and resorption by an effect on the tumour . 5 . It is concluded that prostaglandins may be important in mediating the effects of renal cortical carcinoma and possibly breast cancer on bone destruction . A non-prostaglandin mechanism may also contribute to bone destruction by breast carcinoma.

Ann Intern Med, 1980 Mar, 92(3), 396 - 405
Pemphigus: current concepts; {The maintenance of insulin secretory response of long-term cultured pancreatic islets to glucose et al.; Tsutou A, Kawaguchi A, Taniguchi H, Yoshioka M, Tamagawa M, Seki M, Murakami K, Kobayashi T, Baba S.

The present study was performed to investigate whether long-term cultured rat pancreatic islets possess a postcultural insulin secretory response to hormones and neurotransmitters in spite of their lack of stimulation during the culture period . We also investigated the method of maintaining the insulin secretory response of islets cultured in a physiological concentration of glucose . The tissue culture media were TCM 199 supplemented with 5.5 mM glucose (A medium), 5.5 mM glucose plus 1 mM adenosine (B medium), 16.7 mM glucose (C medium), and 16.7 mM glucose plus 1 mM adenosine (D medium) . Short-term incubation after the culture period of 14 days showed that the islets cultured in B, C and D media maintained the same insulin secretory responsiveness to 8.3 mM glucose and/or 5 microM acetylcholine and also to 1 microM epinephrine as did non-cultured islets . A similar response was found among the islets maintained in B, C and D media . An insulin secretory response to epinephrine and phentolamine was deficient in islets cultured in A medium, whereas it was maintained in those cultured in C medium . The responsiveness of the islets cultured in C medium to the concomitant stimulation by epinephrine and phentolamine was not different from that of the non-cultured islets . It was thus concluded that the addition of adenosine in the culture medium containing the physiological concentration of glucose was as effective in amintaining the insulin secretory ability of the islets as was the culture medium containing a high concentration of glucose, and it was suggested that even the pancreatic islets cultured in these media, though separated from the innervation might preserve acetylcholine and adrenergic receptors similar to freshly isolated islets . Considering the action of adenosine, the necessity of enhancing ATP and C-AMP concentrations in B cells was also suggested in order to maintain the insulin secretory ability of cultured pancreatic islets.

Am J Obstet Gynecol, 1980 Feb 15, 136(4), 419 - 25
Progesterone-induced glycogen accumulation in human endometrium during organ culture; Shapiro SS et al.; An organ culture system was used to evaluate the response of human proliferative endometrium to progesterone stimulation . The glycogen content of proliferative tissue was increased as much as thirteenfold during organ culture in media containing progesterone . The steroid-induced increase in tissue glycogen was detectable after 16 hours and reached a maximum at 48 and 72 hours of culture . Progesterone induced a significant increase in glycogen at a media concentration of 1.6 x 10(-8)M and a maximal increase at 3.2 x 10(-7) to 3.2 x 10(-6)M . At higher media concentrations of progesterone (1.6 x 10(-5)M), glycogen levels failed to reach the maximum obtainable . The extent of the response correlated poorly with the initial glycogen content of the tissue and not at all with the initial content of high-affinity progesterone-binding sites in cytosol . Addition of estradiol-17 beta to medium (10(-10)M to 10(-7)) has no effect on the progesterone-induced increase in tissue glycogen . Delaying the addition of progesterone to the cultures for 24 hours resulted in a diminished glycogen response; the reduced response may be related to a rapid decrease in high-affinity progesterone-binding sites as measured in cytosol prepared from tissues cultured in the absence of progesterone . High-affinity progesterone-binding sites in endometrial cytosol were found to decrease rapidly during the first 24 hours of culture . The addition of cycloheximide or actinomycin D to the culture media inhibited the increase in tissue glycogen caused by progesterone . These results demonstrate that progesterone can induce an in vitro response in human proliferative endometrium similar to that seen in vivo . The response of the endometrium is reproducible and allows for comparisons between grouped data obtained by using tissues from several different donors.

Cell Biol Int Rep, 1980 Feb, 4(2), 185 - 93
Spermine oxidation products are selectively toxic to fibroblasts in cultures of normal human prostatic epithelium; Webber MM et al.; Contamination with fibroblasts has been a major problem in the isolation and establishment of pure cultures of prostatic epithelium, when developing in vitro cell models for studies on carcinogenesis . Addition of spermine to culture media containing fetal bovine serum results in oxidation of spermine to unstable aminoaldehydes which decompose releasing stable acrolein . This oxidation is catalyzed by amine oxidase present in ruminant serum . Prostatic epithelial cells are resistant to these oxidation products at the levels of spermine used . However, these oxidation products are selectively toxic to normal prostatic fibroblasts, which can be eliminated from mixed cultures, and pure cultures of prostatic epithelial cells can thus be established.

J Clin Invest, 1980 Feb, 65(2), 268 - 76
Alveolar macrophage-derived chemotactic factor: kinetics of in vitro production and partial characterization; Merrill WW et al.; Alveolar macrophages are the initial phagocytic cells that encounter foreign material and particulates deposited in the terminal airways . We have examined a mechanism by which these cells, after phagocytic challenge, may control or amplify the inflammatory response in lung parenchyma . Normal human alveolar macrophages (AM) were studied from eight subjects . With in vitro culture, AM produced and released two substances into culture media which have potent chemoattractant activity for blood polymorphonuclear granulocytes (PMN) and negligible activity for mononuclear cells . Release of these factors is maximally stimulated by aggregated human immunoglobulin (Ig)G or zymosan particles; however, simple adhesion of the macrophages to plastic surfaces is also sufficient to stimulate release of these chemotactic substances . The larger substance (10,000 daltons) is immunologically distinct from C5a and interacts with a different PMN membrane receptor than that known to exist for formyl-methionyl-leucyl-phenylalanine . Its chemotactic activity is sensitive to the enzymatic effect of trypsin . Although producing a single elution peak on gelfiltration chromatography, electrofocusing in polyacrylamide gels yielded five peaks of radioactivity . Chemotactic activity was localized to a fraction with a pI = 5.0 . The smaller molecular weight substance has been less well characterized . Thus, the human AM can produce at least two factors which attract PMN and this capability may augment the local inflammatory response in the lung.

J Clin Endocrinol Metab, 1980 Feb, 50(2), 234 - 9
Ectopic production of pregnancy-specific beta 1-glycoprotein by a nontrophoblastic tumor in vitro; Azer PC et al.; To demonstrate the ectopic production of pregnancy-specific beta 1-glycoprotein (PSbetaG) by a nontrophoblastic tumor, the in vitro secretion of this glycoprotein was evaluated in an hCG-producing ovarian cystadenocarcinoma cell line maintained in long term cell culture . Parallelism was demonstrated between the immunoreactive material present in the tissue culture media and highly purified PSbetaG measured by a sensitive and specific RIA . The immunoreactive material was shown to cochromatograph with purified PSbetaG present in normal term pregnancy serum on a Sephadex G-150 column . The tumor PSbetaG was adsorbed to concanavalin A-Sepharose and eluted with alpha-D-methyl-glucoside in a manner similar to purified PSbetaG . The indirect immunoperoxidase method with anti-PSbetaG sera localized PSbetaG in secretory vesicles and in the endoplasmic reticulum of the tumor cells by light and electron microscopy . The addition of sodium butyrate to the tissue culture media stimulated PSbetaG production in a fashion quantitatively similar to that seen with hCG . The number of PSbetaG-staining intracellular vesicles also increased after exposure to butyrate . These studies provide direct evidence for the ectopic production of PSbetaG by a nontrophoblastic tumor cell line.

Cancer Res, 1980 Feb, 40(2), 325 - 8
Procollagens as markers for the cell of origin of human bone tumors; Stern R et al.; Cells derived from osteogenic sarcomas and from Ewing's sarcomas, two malignant bone tumors, were examined for the types of collagens they elaborated into the tissue culture media . Type I procollagen was the predominant species from all osteogenic sarcoma cell lines, a finding consistent with bone cell origin . The Ewing's sarcoma cells contained a prominent peak of type III procollagen and resembled the profile of vascular smooth muscle cells . Fibroblasts derived from skin biopsies taken from amputation specimens synthesized both type I and type III procollagens at the expected ratio of approximately 3:1 . The examination of matrix proteins may provide a general classification scheme for human sarcomas and permit distinction of one tumor from another, as well as from normal fibroblasts.

J Bacteriol, 1980 Feb, 141(2), 687 - 93
Cyanophycin granule polypeptide formation and degradation in the cyanobacterium Aphanocapsa 6308; Allen MM et al.; The effect of a number of conditions on the amount of cyanophycin granule polypeptide {multi-L-arginyl poly(L-aspartic acid)} formed in the unicellular cyanobacterium Aphanocapsa 6308 was determined . Light, CO2, sulfur, and phosphorus starvation as well as the addition of arginine to culture media increased the amount of cyanophycin granule polypeptide in cells when compared with that in cells grown under conditions optimal for growth . Nitrogen limitation and reduction of growth temperature to 30 degrees C decreased the amount of cyanophycin granule polypeptide on a dry-weight basis . Shift-up and shift-down experiments suggest cyanophycin granule polypeptide may be a reserve nitrogen polymer in Aphanocapsa 6308.

J Cell Physiol, 1980 Feb, 102(2), 267 - 77
Sarcoma growth factor (SGF): specific binding to epidermal growth factor (EGF) membrane receptors; De Larco JE et al.; Cells transformed by murine sarcoma viruses (MSV) produce and release into their tissue culture media several polypeptide growth stimulating factors . One of these has been partially purified using Bio-Gel P-60 column chromatography followed by DEAE-cellulose chromatography . This growth factor was assigned the name sarcoma growth factor (SGF), and is here shown to require the epidermal growth factor (EGF) receptor in order to function as a growth factor . DEAE-cellulose chromatography yielded a product that was several-fold purer than the material present in the Bio-Gel P-60 column pool II . The biologically active material from the DEAE-cellulose column, when labeled with 125I, showed specific binding to EGF membrane receptors . The specific binding could be prevented with the addition of either unlabeled EGF or SGF . Both radiolabeled SGF and EGF will bind to live or fixed cells . We were able to bind 125I-SGF as well as 125I-EGF to fixed cells and elute the bound material from fixed receptors . The eluted SGF showed a greater than 25-fold increase in specific binding . The biological activities of EGF and SGF could be bound to and eluted from fixed receptors . The eluted SGF showed a greater than 25-fold increase in specific binding . The biological activities of EGF and SGF could be bound to and eluted from fixed cells . A 3T3 clone lacking EGF receptors was unable to respond to either EGF or SGF, whereas it responded well to serum and several other purified growth factors . The SGF isolated using DEAE-cellulose chromatography was unable to compete in a radioimmune assay using 125I-EGF and antibody to purified mouse submaxillary gland EGF; it also was not precipitated by anti-EGF antibody . From these studies it appears that the SGF produced and released by these MSV-transformed cells combines with and requires the EGF receptor in order to exert its biological effects . The peptide, however, is antigenically distinct from mouse submaxillary gland EGF.

Jpn J Physiol, 1980, 30(5), 767 - 74
Enhancement of erythroid colony formation in vitro by spleen extract from irradiated rats; Kasai S et al.; The effect of spleen extract from irradiated rats on CFU-e and BFU-e colony formation of rat bone marrow cells was investigated by using modified plasma clot culture media . In the presence of erythropoietin (Ep), CFU-e colony formation peaked at 48 hr of culture, and the Ep-induced increase of CFU-e colonies was dose-dependent . The addition of spleen extract enhanced the colony formation more than two-fold in the Ep-containing culture . BFU-e colony formation was also enhanced by the addition of spleen extract . These results indicate that spleen extract from irradiated rats contains factor(s) which stimulates the proliferation of erythroid progenitors.

Zentralbl Gynakol, 1980, 102(11), 592 - 5
{Screening method for diagnosis of bacterial infections of the vagina (author's transl)}; Spitzbart H et al.; Reported is the use of TTC-containing culture media . They were used by the authors in screening tests to determine bacterial vaginal flora . The method seems to be promising in terms of better diagnostic and therapeutic results.

Folia Parasitol (Praha), 1980, 27(4), 349 - 52
Effects of some sugars, amino acids, glycerol and alpha-glycerophosphate on oxygen uptake of some digenetic trematodes; Nizami WA et al.; Effect of various substrates on the oxygen uptake of some digenetic trematodes -- Srivastavaia indica, Gastrothylax crumenifer, Gigantocotyle explanatum from the buffalo, Bubalus bubalis and Isoparorchis hypselobagri from the fish Wallago attu was studied . In all the four species, the effect of various substrates except maltose was found to be stimulatory . The inhibitory or stimulatory effect of these substrates on oxygen uptake reflects the importance of these compounds in the energy metabolism . In spite of the fact that these trematodes were subjected to similar concentration of substrates, the percentage change in oxygen uptake is different in each of the trematodes, the differential utilization is probably related to the different ecophysiological conditions of the habitat of these trematodes . Such studies can be useful in devising better in vitro culture media for these trematodes.

Adv Exp Med Biol, 1980, 128, 635 - 43
Effect of uremic sera on parathyroid hormone (PTH) mediated release of calcium from normal rat embryonal bone maintained in tissue culture; Letteri JM et al.; 1 . 45Ca released from embryonal fetal rat bone into a tissue culture system containing uremic serum was lower than 45Ca measured in culture media containing normal sera . 2 . With stimulation of bone calcium mobilization by the addition of PTH, 1,25(OH)2D3, 24,25(OH)2D3 and 25 (OH)2D3, the 45Ca released into media containing uremic serum was significantly lower than measured in cultures containing normal serum . 3 . Addition of PTH in combination with 24,25(OH)2D3 or with 24,25(OH)2D3 and 1,25(OH)2D3 to bone cultures containing uremic serum increased the 45Ca released into the media when compared to cultures containing uremic serum and PTH.

Acta Anat (Basel), 1980, 106(1), 129 - 40
Differentiative ability of the tibial periosteum for the embryonic chick; Scott-Savage P et al.; The claim that the hypertrophic cartilage of a transversely divided 9-day chick tibia is able to induce the differentiation of osteoblasts when placed against the fibrous periosteum of a second, intact, 9-day tibia in vitro, is refuted . Tibia from 7- and 9-day-old chick embryos were transversely sectioned though either the diaphysis or epiphysis and these diaphyseal and epiphyseal grafts were then placed in close association with the fibrous periostea of intact, host tibiae of the same or different ages, at either the diaphyseal or epiphyseal level . All of the sixteen possible combinations of paired tibiae were established in organ culture for 7 days on three types of culture media and two types of atmospheres . In response to the close contact established when diaphyseal grafts were used, the fibrous layer of the periosteum underwent hyperplasia to form fibroblasts at the contact site . Contact of the graft with the periosteum as not sufficient to allow osteoprogenitor cells to accumulate, to differentiate into osteoblasts, or to deposit osteoid . We concluded that the outer, fibrous layer of the periosteum is fibroblastic and not osteblastic.

Med Interne, 1980 Jan-Mar, 18(1), 75 - 82
HBs antigen and blood glucose concentration; Lenkei R et al.; HBsAg presence was studied by counterelectrophoresis (CEP) in 76 patients with liver cirrhosis and in 431 patients with diabetes mellitus . A striking correlation was found between high blood glucose values and HBsAg absence . Thus, HBsAg-negative cirrhotics (53%) had significantly higher levels of glycemia than the HBsAg-positive patients of the same age group, i.e . 95.75 +/- 6.36 ng/100 ml compared to 78.30 +/- 10.2/100 ml . This absence of HBsAg was also observed in all diabetics but one . As the incidence of HBsAg (CEP) was found to be of 3.63% in 253,460 subjects from different areas of Romania and 6.84% in 14,690 subjects with various non-hepatic diseases included, the chance of finding the 0.2% HBsAg incidence observed in the diabetics would be less than 0.0002 and 0.0001, respectively . The serum HBsAg absence in cirrhotics with high glycemia and in diabetics strongly incriminates the constant high concentrations of blood glucose as the main factor responsible for this negativity . The effect may be direct on virus replication, or indirect, by metabolically-induced hepatic dysfunction interfering with HBsAg secretion or excretion . The presence of high concentrations of glucose in cell culture media might explain the repeated failure of hepatitis B virus serial passage in tissue or organ culture.

In Vitro, 1980 Jan, 16(1), 87 - 92
Plant tumor reversal associated with the loss of foreign DNA; Yang FM et al.; Transformation of plant tissues into crown gall tumors has been associated with the transfer of a portion of a tumor-inducing plasmid (Ti-plasmid) into plant DNA . Various laboratories have regenerated normal-appearing plants from a number of crown gall tumors . This study investigates the fate of the foreign DNA in a series of tissues derived from various parts of a plant regenerated from the tumor BT-37 by Braum and his coworkers . It was found that all the foreign DNA sequences were lost from tissues that had lost all their tumorous traits; whereas the plasmid DNA sequences were still present in tissues that appeared normal but still exhibited tumorous traits when returned to tissue culture media . From these studies it would appear that the presence of the Ti-plasmid sequences in the plant DNA is required for the maintenance of the transformed state.

Biochimie, 1980, 62(1), 27 - 32
In vitro conversion of porcine thyroid cells growing in monolayer into functional follicular cells; Fayet G et al.; Porcine thyroid cells in primary cultures form either monolayers or, when they are stimulated by thyrotropin (TSH), follicles . This system, monolayer-follicle associated cell culture, determines two morphofunctional states of the thyroid cell in vitro . These cells divide, when grown in monolayer . In this article we describe the precise conditions which allow the conversion of monolayer cells into follicles . The rate of cell growth was lowered using a serum-free medium . Cells were concentrated, stimulated by TSH and cultured on poly-L-lysine pretreated plastic flasks . Light and electron microscope studies show that cells reorganize into follicles containing thyroglobulin . Active iodide transport by the cells, as well as detection of thyroid hormones in the cell culture media, demonstrate that these follicles are functional . Formation of monolayers from follicles is in vitro a spontaneous phenomenon . It is linked to the loss of cell polarity, iodide transport, synthesis of hormones and to the decrease of the number of TSH receptor-sites . These main characteristics of differentiation may be regained in vitro after conversion of monolayer thyroid cells into active follicles up to at least generation five.

Cancer Res, 1980 Jan, 40(1), 29 - 35
An inhibitor of lymphocyte proliferation produced by a human colonic adenocarcinoma cell line in culture; Whitehead JS et al.; The culture fluid taken from a human colonic adenocarcinoma cell line, SKCO-1, inhibited mitogenic stimulation of mouse lymphocytes measured by reductions in blast information or {3H}thymidine uptake . The inhibitor was not cytotoxic to lymphocytes nor did it alter the growth rates of 21 other cell lines examined . However, the in vitro growth rates of two murine lymphoid tumor lines were also markedly reduced by the SKCO-1 medium . The tumor cell culture media could be added up to 40 hr after concanavalin A stimulation and still effect complete inhibition of {3H}thymidine uptake . A similar inhibition was observed for phytohemagglutinin (from red kidney bean), lipopolysaccharide, and mixed lymphocyte response assays . The inhibitor has a molecular weight of greater than 100,000 and is stable to heat and ultraviolet radiation, but is destroyed by mild periodate oxidation . The inhibitor may play a role in immunosuppression.

J Supramol Struct, 1980, 13(3), 281 - 94
Cell cycle dependent rate of labelling of cellular and secreted glycosaminoglycans in mouse embryonic fibroblasts; Otto AM et al.; Cultures of embryonic fibroblasts from Balb/c or CBA/J mice were given 12-h pulses of 14C-galactose, or were double-labelled with 3H-galactose and 35H-sulfate . The time course of the rates of labelling of glycosaminoglycans--galactose label was found in the uronic acid moiety--was studied in synchronously and asynchronously growing cultures . Partial synchrony was achieved by trypsinising quiescent, confluent cells and subsequent transfer of cells to new cultures with fresh medium . Synchrony was monitored by measurement of thymidine uptake in parallel cultures . The distribution of label in the hyaluronic acid, chondroitin sulfate, and heparan sulfate fractions from cells and culture media was determined at each time point . Peaks of DNA synthesis were accompanied by or followed 12 h later by a maximal rate of labelling with galactose of secreted glycosaminoglycans, and with the exception of hyaluronic acid--also of cellular glycosaminoglycans . The rate of labelling with galactose of glycosphingolipids in parallel cultures followed a different time course . In double-label experiments the rates of labelling of glycosaminoglycan sulfates with 3H-galactose and 35S-sulfate did not go parallel . In older, quiescent cultures the labelling rate with galactose decreased while the sulfation rate increased . It is discussed that the labelling rate with galactose is indicative of the biosynthetic rate of the glycosaminoglycans . The conclusion is reached that glycosaminoglycans are preferentially synthesized and secreted after the S phase of the cell cycle.

Zentralbl Bakteriol Naturwiss, 1980, 135(4), 367 - 73
{Studies of the nitrogen and carbon content of culture media and of mushrooms, as well as the free amino acid composition of myecelia and fruiting bodies of Lentinus edodes}; Kleinmann-Klar D; The level contents of nitrogen and carbon in the analysed nature substances may be considered the reason for the good growth of Lentinus edodes on this culture medium (sugar-beet molasses and sawdust) . The separate parts of fruit-bodies are showing differences in contents of nitrogen, carbon and free amino-acids; the gills have the highest, the stalk the lowest concentration, that of the hats is taking a middle position . Young and old mycelia as well as the separate parts of fruit bodies are different out of total concentration also in composition of free amino acids . This may be a direction to development and transformations during the change from the vegetative to the reproductive phase.

Cytogenet Cell Genet, 1980, 26(2-4), 165 - 74
Structure of the mitotic apparatus and chromosomes after hypotonic treatment of mammalian cells in vitro; Brinkley BR et al.; Cultured mammalian cells were exposed briefly to culture media made hypotonic with various dilutions of distilled water . Electron microscopic observations and tubulin immunofluorescence were carried out on mitotic cells that had been briefly exposed to hypotonic conditions . Similar observations were made on cells that had been exposed to hypotonic medium and then returned to isotonic conditions . These experiments provided new insight into the structure and physical properties of the mitotic apparatus and chromosomes . Spindle microtubules were reversibly depolymerized by hypotonic treatments . Spindle fibers disappeared after a 15-min exposure to hypotonic medium, leaving only two brightly fluorescent spindle poles . When hypotonically swollen cells were returned to isotonic medium for 15 min prior to fixation, the spindle was reassembled . Several types of spindle aberrations were noted in cells recovering from hypotonic treatment . Chromosomes were greatly expanded by hypotonic treatment, and the kinetochores were disrupted . When treated cells were returned to isotonic culture medium, the chromosomes recondensed and the kinetochores reappeared . Brief hypotonic treatments, described herein, had no adverse effect on cell viability and may prove useful in investigations of the structure and physical properties of the mitotic apparatus.

Int Arch Allergy Appl Immunol, 1980, 63(2), 129 - 38
Human IgE, IgG and IgE antibody synthesis in vitro; Ohta K et al.; Human IgE biosynthesis in vitro was studied by co-culturing T and B cells from atopic patients and normal controls in the presence of 10 microliter/ml pokeweed mitogen for 7 days . IgE and anti-mite IgE ab in the culture media significantly correlated with those in plasma . Normal T cells suppressed in vitro production of IgE and IgE ab significantly more than did atopic T cells but no difference was found in IgG production . The culture of B cells alone (T-depleted fraction) produced IgE equally as well as the co-culture of T and B cells . Anti-mite IgE ab was also produced by B cells alone from mite-sensitive patients . However, IgE biosynthesis in vitro was not consistently affected by pokeweed mitogen . The results suggest a deficiency of the T suppressor system in atopy and the existence of T-independent IgE-producing cells.

Immunobiology, 1980 Jan, 156(4-5), 364 - 71
Human primary T-cell lines in lectin-free media; Fabricius HA et al.; A method of establishing human T-cell lines in tissue culture media containing TCGF but not PHA is described . PBL, initially stimulated by PHA to produce TCGF, continue production for at least 48 hrs after the lectin has been washed off . The TCGF-conditioned medium initiates blast formation of T-cells from freshly isolated PBL and supports indefinite growth of the T-cell blasts from the primary PBL cultures and also from blast-cells obtained from clonally derived T-cell colonies grown in soft agar culture . The established cell lines are identified by cytochemical and biological means as belonging to the T-cell compartment and express spontaneous cytotoxicity against HeLa-cells . The continuously growing T-cells are unable to produce TCGF and depend strictly on external supply of the growth factor . PHA by itself does not support T-cell growth in spite of its ability to elicit a mitogenic response in PBL-cultures . Thus two types of cells must be involved in the mitogenic event: (i) a TCGF-producer and (ii) a TCGF-responder . PHA elicits in the former TCGF production and then TCGF in the later the mitogenic T-cell response . Therefore, not PHA but TCGF might be considered to be the T-cell mitogen.

Dev Biol Stand, 1980, 45, 163 - 73
Replacement of animals in manufacture and control of vaccines; Hennessen W; The first vaccines were obtained from animals infected with the vaccine viruses . Bacterial vaccines were derived from culture media prepared from animal tissues . In some cases the principles of manufacture remained unchanged for more than a century, the methods only were refined . Animal cell culture was not introduced into vaccine production until the second half of of this century . After more than 25 years the search for non-animal substrates and their improvement still goes on . Control of vaccines lagged behind this progress . After a period of no control for the early vaccines, animal tests enabled immunologists to measure safety and potency, thus eliminating the disasters attributable to unsafe products or failures in protection . In vitro methods of control led to a better understanding of the immunology of vaccines . Replacement of animals in manufacture is now to be paralleled by their replacement in control . This will require courage from both the manufacturer and the controller.

J Virol Methods, 1980, 1(4), 213 - 21
Efficient production of mammalian RNA tumor viruses in serum-free culture medium allows rapid RNA subunit purification; Strauss EM et al.; A wide variety of infected mammalian cell cultures were observed to produce high levels of RNA tumor virus particles in the absence of serum for at least 12 h . Virus production was measured by yields of 50-70 S virus RNAs isolated directly from serum-free culture media by chromatography on oligo(dT)-cellulose . Yields of RNAs from viruses produced in serum-free medium were comparable to yields obtained from purified viruses produced in serum-containing medium . Subunits of viral RNAs were thermally dissociated and separated by a new sedimentation system using sucrose gradients with resolving power (in the relevant size range) equivalent to that obtained with electrophoresis in polyacrylamide gels . RNA subunits isolated directly from serum-free medium appeared to be intact as judged by poly(A) content and resedimentation . The overall approach developed here represents dramatic savings in time and effort over previous ways of producing purified RNA subunits from tumor viruses.

Oncodev Biol Med, 1980, 1(3), 181 - 90
Effect of phorbol esters and hormones on rat hepatoma cells producing alpha-fetoprotein; Kaneko Y et al.; Effect of 12-O-tetradecanoyl-phorbol-13-acetate (TPA) on rat AH66 hepatoma cells was studied with a reference to that of insulin and the epidermal growth factor (EGF) . In a short term cell incubation, TPA and EGF caused an approximately 2-fold increase in the production of alpha-fetoprotein (AFP) and other acid-precipitable materials, while the same concentration of insulin brought a 3-fold increase . In a long term culture using a low serum medium, TPA as well as insulin and EGF caused remarkable proliferation of AH66 cells, but the increase in cell number was not accompanied by a proportional increase in the levels of AFP of the culture media . These biological effects of TPA, insulin and EGF appeared to resemble each other, and subsequent hormone binding studies showed that TPA inhibited 125I-EGF binding to its membrane receptors without affecting 125I-insulin binding . Scatchard analysis of TPA effect on EGF binding indicated that TPA altered the affinity of the membrane receptors for EGF without changing the total number of available receptors per cell . From these data, it is suggested that some of the biological effects of TPA on AH66 cells may result from alterations in the functions of cell membrane.

J Immunol Methods, 1980, 39(4), 335 - 41
Development of a cell line secreting monoclonal antibody specific for concanavalin A and use of that antibody as an affinity immunosorbent for tissue culture media used to support long-term T cell growth; Spurll GM et al.; A new cell line, 71A7, secreting a monoclonal mouse IgG1 antibody specific for the protein concanavalin A (Con A) has been established . The stable and rapidly growing line has been propagated in vitro for several months without loss of secretion of antibody . Affinity purified antibody has been used as an immunoadsorbent to remove Con A from tissue culture supernatants for long-term T cell maintenance.

Immunol Commun, 1980, 9(6), 543 - 57
Antigen induced modulation by shed lymphocyte membrane gangliosides; Correa M et al.; A unique regulatory mechanism has been proposed for ganglioside which functions to immune responses to temporarily restrain B lymphocytes of all specificities and at various levels of differentiation . Utilizing antigen competition protocols, experiments was designed to exploit and extend the antigen induction properties of this modulation . Spleen cell cultures were prepared from TNP-BGG primed mice . In vitro stimulation of the cultures with ovalbumin (OVA) followed 24 hours later by addition of TNP-BGG resulted in only one weak TNP hemolytic plaque responses when compared to control cultures which did not receive OVA . OVA induced competitive effects were absorbed by anti-Thy-1 or anti-ganglioside . Glycolipids could be extracted from the culture media supernatants of experimental and control groups as a ganglioside fraction containing Thy-1 determinants . These molecules, when formulated into liposomes, produced the same modulatory effect as that of media from the competitive culture group . These results support and extend the proposal that glycolipids released by antigen stimulated T cells provide unrestricted modulation of B cells to prevent overload during T cell maturation . This regulatory mechanism prevents direct antigen binding and prepares B cell receptivity for further stimuli which orchestrate their terminal differentiation into plasma cells.

Teratog Carcinog Mutagen, 1980, 1(1), 3 - 13
Metabolism of aflatoxin B1, benzo{a}pyrene, and 1,2-dimethylhydrazine by cultured rat and human colon; Autrup H et al.; A model system for comparing carcinogen metabolism between human and rat colon has been developed . Tissue explants maintained under chemically defined conditions were treated with radioactively labeled carcinogens . After incubation for 24 hours, the binding of radioactive carcinogen to DNA was quantitated . Further, the carcinogen-DNA adducts and carcinogen metabolites released into the culture media were identified . Both human and rat colon activate benzo{a}pyrene (BP), aflatoxin B1 (AFB), and 1,2-dimethylhydrazine (DMH) into chemical species that reacted with cellular macromolecules . When human and rat colons were compared, the metabolism of AFB and DMH was qualitatively similar - the same major carcinogen-DNA adducts and metabolic profile . However, the mean binding levels of DMH and AFB to colonic DNA were higher in rats than in humans . BP-guanine adducts were the major adducts formed by both rat and human colonic DNA . However, BP-adenine adducts were observed in rat colonic DNA but not in human colonic DNA . A positive correlation for the binding of BP and DMH to human DNA of different individuals was observed, but no correlation was found between BP and AFB . The data suggest that similar enzyme systems may be involved in the metabolism of BP and DMH, whereas different enzymes might be involved in the metabolic activation of AFB.

Differentiation, 1980, 17(2), 105 - 9
Fetal rat pancreas in organ culture: effects of serum on the development of the endocrine cells; McEvoy RC; Eighteen-day-old rat fetal pancreas was grown in organ culture for four days on medium consisting of tissue culture Medium 199 and varying concentrations of chicken serum . The glucagon and somatostatin concentration of the explants was decreased when the serum concentration of the medium was reduced from 50 to 10% . There was a further reduction in these hormones when the explants were cultured on Medium 199 alone . Explant insulin content was reduced only when serum was omitted from the medium . A "serum factor" tripeptide was not able to substitute for this serum requirement . Heat-inactivation of the serum resulted in a significant increase in medium insulin content and an increase in both the insulin and glucagon contents of the explants . This increase in hormone content was directly correlated with increases in the beta and alpha cell volumes of the explants . There was no change in the somatostatin content or delta cell volume of the explants grown on heat-inactivated medium . It is suggested that the serum is an important component of the culture media and is apparently required in high concentration for the development of the islet cells in vitro . The islet cell types differ in their requirement for serum . The alpha and delta a higher concentration than do the beta cells.

Thromb Haemost, 1979 Dec 21, 42(4), 1207 - 16
Macrophage inhibition of collagen-induced platelet aggregation; Morland B; Collagen was incubated with cells or media fractions of mouse peritoneal macrophage cultures, and its aggregating effect on human platelets was tested . Incubation with lysates of cultured cells completely abolished the normal collagen-induced platelet aggregation, while incubation with media fractions only caused partial inhibition . The latter inhibition was more pronounced after macrophage phagocytosis of latex particles, while endocytosis of endotoxin had no effect . Corresponding macrophage cultures were also tested for specific collagenase activity, using 14C-glycine labelled collagen as substrate . Collagenase activity was found in the culture media fractions only, and the enzyme activity could be enhanced by endocytosis of latex as well as endotoxin . It appears that the effect of macrophage lysates and media on collagen-platelet interaction cannot be ascribed only to secretion of collagenase from macrophages.

Experientia, 1979 Dec 15, 35(12), 1060 - 1
Epidermal growth factor stimulates proliferation of rat hepatoma cells producing alpha-fetoprotein; Kaneko Y et al.; Epidermal growth factor stimulated both {3H}thymidine uptake and proliferation of rat AH66 hepatoma cells . However, the increase in cell number was not accompanied by a proportional increase in the levels of alpha-fetoprotein of the culture media . The effects of EGF on the cell proliferation were antagonized by N6,O2'-dibutyryl cAMP.

J Clin Pathol, 1979 Dec, 32(12), 1226 - 7
Pseudo-leptospires in blood culture; Rahman M et al.; Spiral filaments seen in the blood cultures of two patients with fever and jaundice were initially thought to be leptospires; these were later proved to be artefacts . An investigation was carried out to exclude the possibility of laboratory contamination of the culture media and to find out how these objects were produced . The significance of the findings is discussed in relation to the possibility of a mistaken diagnosis in routine laboratories which have a limited experience of leptospires.

In Vitro, 1979 Dec, 15(12), 1023 - 31
Adult rat lung in organ culture: maintenance of histotypic structure and ability to synthesize phospholipid; Weinhold PA et al.; The maintenance of adult rat lung explants in organ culture was assessed both morphologically and biochemically . A comparison of several culture media indicated that Ham's F12K plus 1.0 microM dexamethasone, which maintained the explants for 14 days, was superior . The ability of the explants to synthesize dipalmitoylphosphatidylcholine increased with the length of cultivation to values greater than the noncultivated controls . The DNA content remained constant for 7 days, and a relatively normal structural relationship between type I and type II pneumocytes was maintained . Explants cultivated in Ham's F12K without dexamethasone did not maintain a histotypic morphology; the type II pneumocytes appeared to proliferate and the ability to synthesize phosphatidylcholine decreased.

Appl Environ Microbiol, 1979 Dec, 38(6), 1052 - 5
Cyclopiazonic acid production by Penicillium camemberti Thom and natural occurrence of this mycotoxin in cheese; Le Bars J; Every Penicillium camemberti strain freshly isolated from 20 commercial cheese brands produced cyclopiazonic acid in two culture media at 25, 13, and 4 degrees C; the toxin yield was greatly dependent on the strain and environmental parameters (medium, temperature, and incubation time) . The toxigenic ability appeared as a log-normal distribution . This mycotoxin was found in the crust (0.05 to 0.1 microgram/g in three samples, 0.1 to 0.2 microgram/g in five samples, and 0.4, 1, and 1.5 microgram/g in three other samples) but not in the inner part . When its acute toxicity is considered, doses eventually ingested by consumers are very low (lower than 4 microgram) . Means for prevention are discussed . A highly toxigenic strength and rate appear to be necessary features leading to natural contamination in cheeses . The distribution of toxigenic ability makes possible without delay a choice of weakly toxic strains.

Am J Vet Res, 1979 Dec, 40(12), 1781 - 2
Gastrointestinal nematode immunization trials in cattle; Herlich H et al.; Three experiments were conducted to immunize calves against Ostertagia ostertagi, Trichostrongylus axei, and T colubriformis . Calves were given intraperitoneal injections with in vitro-grown parasitic larvae of the three species and IV and intraperitoneal injections of exoantigens obtained from culture media used to grow O ostertagi . None of the treatments provided immunity to subsequent oral challenge exposure with normal infective larvae.

Br Med J, 1979 Dec 1, 2(6202), 1392 - 3
Immunoreactive calcitonin in leukaemia; Hillyard CJ et al.; A radioimmunoassay was used to measure concentrations of immunoreactive human calcitonin (HCT) in plasma and leucocytes from patients with various leukaemic and myeloproliferative disorders . Plasma immunoreactive HCT concentrations were increased in 32 out of 33 patients with chronic granulocytic leukaemia (CGL) and in all eight patients with acute myeloid leukamia (AML) at presentation or in relapse . Out of 11 patients with other myeloproliferative disorders, eight had increased plasma immunoreactive HCT concentrations . Buffy-coat-cell extracts and culture media from peripheral leucocytes of patients with CGL also contained increased immunoreactive HCT concentrations . In contrast, plasma from patients with chronic lymphocytic leukaemia, acute lymphoblastic leukaemia, and AML in remission had low or undetectable immunoreactive HCT concentrations . Increased plasma and cellular concentrations of immunoreactive HCT may be a consequence of abnormal proliferation of myeloid cells and might prove to be valuable in predicting relapse in patients with myeloid leukaemias.

Aust N Z J Surg, 1979 Dec, 49(6), 716 - 20
Transplantation of organ cultured rat parathyroids; Gough IR et al.; The hypothesis that tissue culture alters the immunogenicity of grafts by removal or inactivation of passenger leucocytes has been investigated in an inbred rat parathyroid allograft model . DA rat (Ag-B4) parathyroid glands cultured for up to three weeks in Eagle's minimal essential medium (MEM) alone, or MEM with added donor-specific antilymphocyte serum, did not survive longer than fresh glands when allografted into Lewis rat (Ag-B1) recipients . Subsequent in vitro comparisons of three different culture media established that RPMI 1640 was superior to MEM and Medium 199 for rat parathyroids . However, further series of transplants after culture in RPMI 1640 alone also failed to produce prolongation of allograft survival.

Am J Clin Pathol, 1979 Dec, 72(6), 977 - 9
Storage and transport of cultures for Herpes simplex virus, type 2; Yeager AS et al.; Tryptic soy broth and brain-heart infusion broth maintained herpes simplex virus within +/- 1 serial dilution of the original titer in all of 51 samples held for one to three days at 4 C . In contrast, holding temperatures of -20 C or 22 C resulted in losses of titer of 10(2) or more . Holding periods as long as five days prior to inoculation did not delay the appearance of the cytopathic effect following inoculation . Unmodified tryptic soy broth or brain-heart infusion broth and holding periods as long as three days at 4 C prior to arrival in the laboratory give as satisfactory results as do traditional viral culture media and prompt inoculation of the specimen.

Zh Mikrobiol Epidemiol Immunobiol, 1979 Dec, (12), 45 - 7
{Suppressing action of splenic cells on macrophage phagocytic activity}; Boglazova EK et al.; The culture media withdrawn from 18-hour cultures of live spleen cells suppressed the phagocytic activity of peritoneal macrophages in mice . The level of suppression, as estimated for an equal number of spleen cells, varied in individual animals, which seemed to be connected with the level of normal infection in the animals . In the process of development of Ehrlich's carcinoma in mice suppressors were produced in the spleen in an increased amount, but only in connection with the total increase in the number of spleen cells, and not due to the selective accumulation of suppressor cells.

Brain Res, 1979 Nov 9, 177(1), 105 - 14
Antiserum induced myelination inhibition in vitro without complement; Dorfman SH et al.; Exposure of neonatal rat cerebellum cultures to antiserum to whole spinal cord or galactocerebroside inhibited myelin formation regardless of whether guinea pig serum was added fresh or after heating to 56 degrees C for 1 h in order to achieve complete removal of hemolytic complement activity . Myelination followed removal of antisera from the culture media . This suggests that the inhibition of primary myelination by anti-CNS tissue antiserum occurs through some mechanism other than as the result of a cytotoxic reaction against oligodendrocytes mediated via the complement system.

Nature, 1979 Nov 8, 282(5735), 207 - 8
Genetic defect in secretion of complement C5 in mice; Ooi YM et al.; A genetic deficiency of the fifth (C5) component of complement1-3, a serum glycoprotein of molecular weight (MW) 220,000 (ref . 4), has been found in 39% of inbred strains of mice3 . Sera of deficient mice lack detectable C5 activity and protein2,3 . In addition deficient mice produce antibody to mouse C5 when injected with sera from C5 sufficient (normal) strains . Levy et al.5 showed that somatic cell hybrids between C5 deficient (B10.D2/old line) macrophages and either C5 sufficient (B10.D2/new line) mouse kidney or chicken erythroblasts secreted haemolytically active mouse C5 in vitro . Several possible molecular mechanisms to account for the findings were considered, but insufficient direct data were available to choose among them . We recently reported that mouse (CD.1 strain) peritoneal cells in culture synthesise and secrete a single chain precursor, pro-C5 (MW approximately 210,000), of the two-chain (alpha chain, 125,000 and beta chain 83,000 MW) C5 protein6 . Radiolabelled precursor C5 was contained within the cells and was secreted into the tissue culture media . Using similar methods, we now find that C5 deficiency in each of five different mouse strains (AKR, SWR, DBA/2J8 A/HeJ and B10.D2/old line) is due to a failure in secretion of C5 protein and not to a failure in biosynthesis of pro-C5.

J Infect Dis, 1979 Nov, 140(5), 798 - 801
Lack of specific effect of adenine arabinoside, human interferon, and ribavirin on in vitro production of hepatitis B surface antigen; Lemon SM et al.; PLC/PRF/5 hepatoma cells continue to produce hepatitis B surface antigen (HBsAg) after greater than 80 passages in vitro, but they do not express other markers of heaptitis B virus (HBV) replication . In this respect, they resemble most liver cells that are persistently infected with HBV . PLC/PRF/5 cells were cultured in the presence of adenine arabinoside, human fibroblast interferon, and ribavirin to determine whether production of HBsAg was sensitive to these antiviral agents . HBsAg released into culture media was detected by radioimmunoassay, and cellular protein synthesis was assessed by {3H}amino acid incorporation studies . A dose-related inhibition of HBsAg occurred with each antiviral agent that was tested, but in each case, this inhibition was matched by a reduction of cellular protein synthesis to a similar degree . Thus, no specific effect on the production of HBsAg was found with any of the antiviral agents tested.

J Lab Clin Med, 1979 Nov, 94(5), 699 - 707
Endogenous erythroid colony-forming cells in fetal and newborn sheep; Roodman GD et al.; In plasma clot cultures fetal, newborn, and adult sheep bone marrow cells respond to Ep, giving rise to large numbers of colonies of erythroid cells . In fetal and newborn, but not adult, sheep the marrow produced EEC in the absence of added Ep . The EEC-forming cells represent a transitory population of erythroid progenitors, persisting for approximately 2 month post-partum . The activity of these cells in the normal newborn sheep displayed a cyclic pattern, which was abolished in the presence of high circulating Ep levels . In both fetal and newborn sheep, EEC formation was inhibited by prior neutralization of Ep present in culture media with anti-Ep, suggesting that as in patients with PV the proliferation and/or differentiation of EEC-forming cells of sheep is subject to Ep control.

Am J Vet Res, 1979 Nov, 40(11), 1649 - 51
Cryopreservation of bovine mononuclear leukocytes; Kleinschuster SJ et al.; A technique of cryopreservation of bovine mononuclear leukocytes for use in lymphocyte stimulation tests and cell identification studies has been developed . Dimethyl sulfoxide was used as the cryopreservation agent . Cells were frozen to --40 C at a controlled rate of --1 C/minute and then to --80 C at a rate of --4 C/minute, and were subsequently stored in liquid nitrogen (--109 C) . The cells were than recovered by rapid thawing in a 37-C water bath, diluted with tissue culture media, washed, and used for lymphocyte stimulation and cell identification tests . The magnitude of the lymphocyte blastogenic responses of frozen and thawed mononuclear leukocytes was similar to that seen with freshly collected cells . Additionally, percentages of T cells, B cells, and monocytes were similar between frozen and thawed cells and freshly collected cells.

J Protozool, 1979 Nov, 26(4), 648 - 52
Five trypanosomatid species of insects distinguished by isoenzymes; Goncalves de Lima VM et al.; Blastocrithidia culicis, Crithidia deanei, Crithidia fasciculata, Herpetomonas samuelpessoai, Leptomonas seymouri and Leishmania tarentolae grown in cultures were compared by electrophoretic mobility for isoenzymes in 6 enzymes . All species were found distinct in these characteristics . Endosymbiotic C . deanei, which was identical to the aposymbiotic C . deanei in 5 enzymes, had an extra band in aspartate aminotransferase . No differences in isoenzymes were found between members of one species maintained in 2 different culture media.

Am J Obstet Gynecol, 1979 Oct 15, 135(4), 499 - 502
In vitro effect of dopamine and pimozide on human chorionic gonadotropin secretion; Macaron C et al.; In human placental explants cultured in vitro, dopamine inhibited human chorionic gonadotropin (hCG) secretion into the culture media . In the control flasks, the level of hCG secretion was 751 +/- 215 mIU/gm of tissue . When 1 mM of dopamine was added, hCG levels decreased to 321 +/- 57.6 mIU/gm of tissue (n = 6, P less than 0.1)--5 and 10 mM of dopamine significantly inhibited hCG secretion . In contrast, 1 mM of pimozide enhanced hCG secretion by 1.8-fold compared to control levels (1,707 +/- 343 versus 3,117 +/- 0.005) . This in vitro effect on hCG is similar to the effect of dopamine and pimozide on hCS secretion by placental explants.

In Vitro, 1979 Oct, 15(10), 751 - 7
Major metabolic pathways and hormone production in unstimulated monolayer cultures of the rat anterior pituitary; Rappay G et al.; In cell cultures derived from anterior pituitary glands of rats, enzyme activities of cell homogenates and hormone (GH, PRL, LH, and FSH) content of the culture media were measured . Sex differences in enzyme activities representing major metabolic pathways (citrate cycles, pentose cycles, and glycolysis) were demonstrated both in freshly dispersed cells and in 8-day-old cultures; in cultures of both sexes enzyme activities increased during cultivation . In cultures derived from female rats, cell protein doubled by the 12th day and remained constant for up to the 24th day in culture, whereas enzyme activities showed changes suggesting that cell metabolism shifted to anaerobic glycolysis during cultivation . In the culture media the presence of four pituitary hormones was demonstrated for as long as 3 weeks of cultivation with variable secretion dynamics; the release of gonadotropic hormones diminished gradually whereas that of GH remained constant and PRL levels increased with time . These results indicate that under strictly defined culture conditions pituitary cells may function in spite of profound metabolic changes.

Immunology, 1979 Oct, 38(2), 257 - 63
The responsiveness of efferent lymph cells to phytohaemagglutinin during the response of the popliteal node to dinitrophenylated bovine serum albumin; English LS; Cells, recovered from the efferent lymphatics of the popliteal nodes of sheep during in vivo responses to dinitrophenylated bovine serum albumin (DNP-BSA), were examined for their responsiveness to phytohaemagglutinin (PHA) in vitro during the first 4--5 days of the immune response . The response was almost totally depressed when cells collected throughout the response were cultured with supra-optimal doses of the mitogen . Optimal and suboptimal doses of PHA resulted in greatly enhanced responses in cells collected in the first two periods (0--6 h; 6--12h) of the response; the presence of high doses of DNP-BSA in the culture media prevented the enhancement in the second period but had no adverse effects on the cells collected during the first 6 h . Efferent cells collected after 12 h generally showed decreased responses, this depression being maximal in cells collected 2--3 days after antigenic stimulation . Later in the response the cells again exhibited enhanced responses . The possible interpretations of these results in terms of regulation of the response are discussed.

J Clin Microbiol, 1979 Oct, 10(4), 586 - 9
Lipoproteins as substitutes for serum in Mycoplasma culture medium; Washburn LR et al.; Crude lipoprotein-containing fractions obtained from sera of three different animal species were tested, in combination with bovine serum in Mycoplasma pneumoniae culture medium . All sera yielded at least one lipoprotein-containing component which was considerably more effective in promoting mycoplasma growth than the unfractionated serum sample from which it was derived . The very low activity of certain whole-serum samples tested in this investigation suggests that toxic substances may be present in whole serum which are not contained in the lipoprotein preparations . The greatest activity appeared in the high-density lipoprotein-containing components of bovine and horse sera and the low-density lipoprotein-containing components of human serum . The high degree of growth-supporting activity of these crude lipoprotein-containing serum components suggests that they may be useful as serum substitutes in mycoplasma culture media.

Acta Pathol Microbiol Scand {C}, 1979 Oct, 87(5), 347 - 52
Isoelectric focusing of macrophage culture media and the effect of the fractions on the synthesis of DNA and collagen by fibroblasts; Jalkanen M et al.; Macromolecules from rat peritoneal macrophage culture media were separated into 30 fractions by flat bed isoelectric focusing (IEF) . The fractions were tested for their influence on thymidine and proline incorporation into cultured rat granuloma fibroblasts . Three fractions, stable after freezing and lyophilization, were of interest: one inhibiting thymidine incorporation (focusing at pH 7.3-7.6), another stimulating thymidine incorporation (focusing at pH 4.4-5.3), and the third stimulating proline incorporation into cells and medium collagen (focusing at pH 10.2-10.7) . The last also exhibited a ribonuclease (RNase) activity with a pH-optimum of 7.0-7.5.

Appl Environ Microbiol, 1979 Sep, 38(3), 412 - 5
Characteristic gamma-lactone odor production of the genus Pityrosporum; Labows JN et al.; Mass spectrometric-gas chromatographic analysis of culture headspaces revealed that members of the genous Pityrosporum produce volatile gamma-lactones during growth on lipid-containing media . Representative members of other yeast genera found on humans failed to produce these compounds . Addition of lecithin, oleic acids, triolein, or human sebum to the culture media stimulated gamma-lactone production by Pityrosporum species . All yeasts tested produced isopentanol and phenylethanol . Production of gamma-lactones may serve as a valuable characteristic in the identification of organisms of the genus Pityrosporum.

Cancer Res, 1979 Sep, 39(9), 3405 - 10
beta-Hexosaminidase isoenzymes in tissues, cultured cells, and media from human fetal intestine and colonic adenocarcinoma; Tsao D et al.; Isoelectric focusing of tissue homogenates revealed a predominance of beta-hexosaminidase B in colonic adenocarcinoma, whereas beta-hexosaminidase A was greater in paired normal-appearing colonic mucosa from the same patients as well as in a specimen of human fetal colonic mucosa . Because of the recognized cellular heterogeneity of these tissues, our studies were extended to an examination of these isoenzymes in 20 cultured epithelial cell lines derived from human fetal intestine and colonic carcinoma as well as the secreted enzymes in their culture media . While the B:A isoenzyme ratio was higher in human cancer cells as compared to fetal cells, some of the cancer cell lines had a greater proportion of the A isoenzyme . Examination of the isoenzyme profiles in the media of these cells revealed a greater B:A ratio whether of fetal or cancer cell origin . These studies parallel the apparent biological differences of neoplastic colonic epithelium occurring in vivo and are reminiscent of reported oncodevelopmental changes in enzymatic properties present in some malignant tissues . The differential stabilities of these two isoenzymes derived from the culture media of both types of cell lines in vitro may limit their value as markers of human colonic neoplasia.

J Biol Chem, 1979 Aug 25, 254(16), 7785 - 94
Stimulation and inhibition of cellular functions by glucocorticoids . Correlations with rapid influences on chromatin structure; Johnson LK et al.; The effect of media conditions on the glucocorticoid response has been examined in three types of cultured cells . In rat pituitary tumor cells (GC cells) growth hormone production was stimulated by glucocorticoids provided fresh culture media was present (enriched media conditions) . In contrast, dexamethasone either failed to induce or deinduce growth hormone synthesis if added to cultures which had not received fresh media for 3 days (depleted media condition) . With human skin fibroblasts, cortisol stimulated {3H}thymidine incorporation in the enriched condition but inhibited this response in the depleted condition . In mouse lymphoma (S49) cells the enriched media conditions significantly delayed the killing response to glucocorticoids (20% killing after 24 h versus 90% killing after 24 h for the depleted condition) . Thus, the magnitude and in some cases, the direction of the glucocorticoid response are sensitive to the conditions to which the cells are exposed . In all three cell types the steroid also rapidly (detectable by 15 min, maximal by 2 h) altered chromatin structure as detected by a change in the number of initiation sites for Escherichia coli RNA polymerase assayed under cell-free conditions . This early nuclear response could be in a positive or negative direction and was also affected by the culture conditions; enriched media favored a positive or less negative effect on the initiation sites by the steroid, while depleted media favored a steroid-induced inhibition of this chromatin function . In S49 and GC cells the kinetics and magnitude of the change in chromatin closely followed receptor . glucocorticoid complex binding to nuclei while removal of dexamethasone from the culture media resulted in a rapid (t 1/2 = approximately 20 min) disappearance of the effect which paralleled loss of bound hormone from the nucleus . The glucocorticoid effect on chromatin was not observed in two lines of glucocorticoid-resistant mutant S49 cells . One line (R-) lacks detectable glucocorticoid receptors; the other line (Nti) has receptors that bind the hormone normally, but the receptor . glucocorticoid complexes bind more avidly to the nucleus . These results suggest that the receptor is involved in both the stimulatory and the inhibitory effects on chromatin . The findings in the Nti cells and of a slight lag between nuclear binding of receptors and initiation site alteration implies that some receptor property, in addition to nuclear binding per se, is responsible for the influence on chromatin . These results are discussed in terms of a model in which steroid hormones initiate their actions by influencing a reaction that modifies chromatin structure . The direction and magnitude of the reaction, and its effect on the expression of specific genes, are dictated by the metabolic state and differentiation of the cell.

Int J Cancer, 1979 Jul 15, 24(1), 45 - 8
Cell lines with spontaneous secretion of pregnancy-associated alpha 2-globulin; Lundgren E et al.; Pregnancy-associated alpha 2-globulin (PA alpha 2G) was quantitated by radioimmunoassay in culture media from 34 exponentially growing human cell lines . Only 8/34 cell lines produced PA alpha 2G . The highest secretion was found in cell lines of histiocytic lymphoma origin while it was low in some carcinoma-derived lines . The results may support an assumption that PA alpha 2G is a normal cell product of B-lymphoid, monocytic and epithelial cells but may also simply indicate that ectopic PA alpha 2G production is especially common in tumors derived from such cells . The availability of PA alpha 2G-producing cell lines should facilitate studies of the immunoregulatory role of this protein.

Cancer Res, 1979 Jul, 39(7 Pt 1), 2711 - 7
Immunological activity of regional lymph nodes in tumor-bearing mice; Barna BP et al.; Regional (popliteal) lymph node cells (RLNC) from A/Jax mice, inoculated in each hind foot with isogeneic Sarcoma 1 tumor cells, demonstrated cytotoxicity in vitro at Day 14 of tumor growth but lost this ability by Day 21 . These noncytotoxic RLNC were capable of suppressing activity of other cytotoxic lymphoid cells, but after incubation for 24 hr in vitro their cytotoxicity was restored and their suppressive activity was abrogated . RLNC responsible for cytotoxicity were removed by treatment with anti-theta serum plus complement . The suppressive effect of RLNC was found to be mediated by a soluble "blocking" factor which was released into the culture medium after 24 hr incubation . The factor was not detected in culture media from RLNC pretreated with anti-theta serum plus complement prior to incubation . Absorption of RLNC culture supernatants with tumor-bearer spleen cells, but not with normal spleen cells, completely removed the blocking factor, while absorption by Sarcoma 1 cells significantly reduced blocking activity . The factor was trypsin sensitive, was retained on an Amicon XM100 filter, and did not demonstrate the presence of antibody to Sarcoma 1 in a radioimmunoassay . Although the exact nature of the factor has not been established, it appears to be a receptor antigen complex from T-cells of tumor-bearing animals.

C R Seances Acad Sci D, 1979 Jun 18, 288(22), 1687 - 90
{Calcifying bacteria and carbonic anhydrase}; Billy C et al.; Carbonic anhydrase activity was measured on bacteria known for their calcifying power . The results obtained allow us to conclude that carbonic anhydrase is not present in their enzymatic equipment . Nevertheless, the addition of carbonic anhydrase in culture media increases the growth of the cultures and their calcifying power . This result is due to the direct action of the enzyme on the pH of the culture medium . The results obtained could be applied to the mechanism of dental plaque calcification.

Pflugers Arch, 1979 Jun 12, 380(2), 165 - 70
Action potentials can propagate along small strands of smooth muscle; Barr L et al.; Bundles of taenia coli muscle as small as 25 micrometers in diameter were dissected from guinea pigs and incubated in an organ culture media for several days . We found that use of an organ-culture bathing solution greatly extended the in vitro responsiveness as well as survival of such small bundles . Electronmicrographs showed that surviving strands of cells constituted only a very small fraction of the cross section of these bundles . Nonetheless, still they supported propagating nondecrementing action potentials . This means that in some cases strands, only a few cells in cross section, supported propagating action potentials . The long length constant and parallel orientation of cells provide a basis assuming cells act as parallel core conductor segments . For these reasons we have called into question the notion that for propagation to occur there must be a facilitating cooperation between large numbers of smooth muscle cells in parallel . Indeed we suggest that the limiting size for propagation is a strand of single cells.

Biochim Biophys Acta, 1979 Jun 12, 585(2), 165 - 87
Synthesis of the calcium transport ATPase of sarcoplasmic reticulum and other muscle proteins during development of muscles cells in vivo and in vitro; Ha DB et al.; The effect of medium Ca2+ concentration upon the concentration and the rate of synthesis of muscle proteins was investigated in chicken pectoralis muscle cultures . There is an easily identifiable class of muscle protein which includes the Ca2+-ATPase of sarcoplasmic reticulum, myosin, troponin C, ATP : creatine phosphotransferase, muscle specific actin, tropomysin 1 and 2, and muscle hemagglutinin, which show a large increase in concentration during normal development . The increased synthesis of these proteins was inhibited, without inhibition of cell proliferation, in culture media of relatively low Ca2+ concentration, 0.05--0.3 mM, where fusion was prevented . Similar medium Ca2+ concentration was required for the expression of all these proteins, suggesting their coordinate regulation . The proteins are denoted as 'calcium-modulated proteins' . The increased Ca2+ transport activity of sarcoplasmic reticulum in cultured chicken pectoralis muscle cells during development at 1.8 mM medium calcium concentration represents de novo synthesis of the Ca2+ transport ATPase, as shown by immunoprecipitation, active site labeling and direct identification of the Ca2+ transport ATPase on two-dimensional gel electropherograms of whole muscle homogenates . The concentration and the turnover rate of the majority of the muscle proteins is not affected significantly by medium Ca2+ concentration between 0.06 and 1.8 mM . It is proposed that increase in cytoplasmic free Ca2+ concentration during fusion plays a central role in the regulation of the synthesis of calcium-modulated proteins.

Prostaglandins, 1979 Jun, 17(6), 905 - 14
Prostaglandin synthesis by fetal rat bone in vitro: evidence for a role of prostacyclin; Raisz LG et al.; Prostaglandin synthesis by fetal rat bones was examined by thin-layer chromatography of culture media after preincubation with labeled arachidonic acid . Cultures in rabbit complement (non-heat inactivated serum) were compared with cultures in heat-inactivated serum or cultures treated with indomethacin . The major complement-dependent products were PGE2, PGF2 alpha and 6-keto-PGF1 alpha, the metabolite of prostacyclin (PGI2) . Since PGI2 had not been previously identified in bone its ability to stimulate bone resorption was tested . Repeated addition of PGI2 stimulated release of previously incorporated 45Ca from fetal rat long bones in both short-term and long-term cultures at concentrations of 10(-5) to 10(-9)M . Because of the short half life of PGI2 in solution at neutral pH, we tested a sulfur analog, thiaprostacyclin (S-PGI2) which was found to be a stimulator of bone resorption at concentrations of 10(-5) to 10(-6)M . These studies suggest that endogenous PGI2 production may play a role in bone metabolism . Since vessels produce PGI2 it is possible that PGI2 release may be responsible for the frequent association between vascular invasion and resorption of bone or calcified cartilage in physiologic remodeling and pathologic osteolysis.

Zentralbl Bakteriol {B}, 1979 May, 168(3-4), 361 - 6
{Lipolytic activity from milk isolated strains of the species Thermoactinomyces vulgaris, Tsiklinsky 1899 (author's transl)}; Falkowski J; Thirty strains of the species Thermoactinomyces vulgaris (Tsiklinsky, 1988) were isolated from milk and tested for their lipolythic activity . After incubation for 120 hours at 55 degrees C they formed distinct diffusion zones on culture media containing tributyrin and tween-80 . In all strains exocellular lipases were found . Their action on the milk fat increased the number of the free fatty acids . Triglycerides, free fatty acids as well as diglycerides and monoglycerides were identified as split products chromatographically . The activity of the endolipases was lower . In four of the most active endolipases cholesterol was identified as split product of milk fat in addition to those mentioned above.

Can J Neurol Sci, 1979 May, 6(2), 241 - 2
Pyruvate dehydrogenase, lipoamide dehydrogenase and citrate synthase activity in fibroblasts from patients with Friedreich's and Charlevoix-Saguenay ataxia; Melancon SB et al.; The activity of lipoamide dehydrogenase and two closely related enzymes was studied simultaneously in early, mild, and late passage fibroblast cultures . Friedreich's ataxia fibroblasts tended to lose pyruvate dehydrogenase and citrate synthase activities, while lipoamide dehydrogenase activity remained constant with aging of the cells . Mean pyruvate dehydrogenase activity was lower over-all in fibroblasts from ataxics . Mean citrate synthase activity was higher in ataxic fibroblasts . Present tissue culture media do not represent the best conditions in which to reproduce cofactor binding defects such as those found in other genetic diseases with structural enzyme mutations.

Zh Mikrobiol Epidemiol Immunobiol, 1979 May, (5), 87 - 90
{Dynamics of Leptospira colony formation on solid nutrient media}; Kiktenko VS et al.; The formation of Leptospira colonies on solid culture media varying in composition were studied . In all species of Leptospira 3 main periods of colony development were revealed, each of them having its characteristic morphological changes connected with the growth of the colony deeply into agar.

Am Fam Physician, 1979 May, 19(5), 121 - 6
Office analysis of bacteriuria; Miriovsky MF; Several techniques for determining significant bacteriuria are convenient, accurate, inexpensive and applicable for office use . The nitrite test, the dehydrated culture media technique, the dip-slide method and the agar tube method are all applicable for screening at-risk populations and for treatment follow-up . In addition, the accuracy, subculturing capabilities and potential for presumptive bacterial diagnosis of the dip-slide and agar tube methods qualify them as excellent office culturing systems.

Mol Cell Endocrinol, 1979 Apr, 14(1), 73 - 9
Follicular fluid stimulation of steroidogenesis in immature granulosa cells in vitro; Ledwitz-Rigby F et al.; Granulosa cells from small (1-2 mm) immature porcine follicles were cultured in monolayer in culture media composed of equal parts of culture medium 199 and either (a) fluid from small follicles, (b) fluid from large (6-12 mm) follicles or (c) adult female porcine serum for 6 days, with or without 100 ng LH and/or 2 microgram FSH/ml . Both basal and gonadotrophin-stimulated progesterone secretion were greater in the presence of fluid from large follicles than in serum, for all 6 days . After 4 days of culture, fluid from small follicles enhanced gonadotrophin-stimulated progesterone secretion over that occurring in serum, but to a lesser extent than fluid from large follicles . These studies suggest the presence of a maturation stimulating molecule(s) in follicular fluid which increases in activity or concentration as the follicles enlarge . This factor may be essential for normal granulosa cell maturation in vivo.

J Infect Dis, 1979 Apr, 139(4), 478 - 82
Enhanced isolation of Mycoplasma pneumoniae from throat washings with a newly-modified culture medium; Tully JG et al.; Two hundred throat washings, previously screened and presumed negative for Mycoplasma pneumoniae in conventional mycoplasma culture media, were retested for the organism in a modified medium (PS-4) initially developed for cultivation of a tick-derived Mycoplasma (spiroplasma) . The organism was rapidly identified with an agar plate immunofluorescence procedure . M . pneumoniae was isolated from 69 (34.5%) of the 200 "negative" specimens cultured on a diphasic SP-4 medium, in contrast to 10 isolations (5%) made on conventional diphasic mycoplasma medium . This enhanced recovery of M . pneumoniae represented a combination of a superior culture medium and a more efficient identification technique . The findings suggest that these procedures might be effectively applied to the recovery of M . pneumoniae from all likely host and that improved recovery of the organism may aid in the interpretation of a number of puzzling questions about the epidemiology of M . pneumoniae infections.

J Cell Biol, 1979 Apr, 81(1), 193 - 214
Immunological studies of the embryonic muscle cell surface . Antiserum to the prefusion myoblast; Friedlander M et al.; Xenogeneic antisera raised in rabbits have been used to detect compositional changes at the cell surfaces of differentiating embryonic chick skeletal muscle . In this report, we present the serological characterization of antiserum (Anti-M-24) against muscle tissue and developmental stage-specific cell surface antigens of the prefusion myoblast . Cells from primary cultures of 12-d-old embryonic chick hindlimb muscle were injected into rabbits, and the resulting antisera were selectively absorbed to obtain immunological specificity . Cytotoxicity and immunohistochemical assays were used to test this antiserum . Absorption with embryonic or adult chick heart, brain, retina, liver, erythrocytes, or skeletal muscle fibroblasts failed to remove all reactivity of Anti-M-24 for myogenic cells at all stages of development . After absorption with embryonic myotubes, however, Anti-M-24 no longer reacted with differentiated myofibers, but did react with prefusion myoblasts . The myoblast surface antigens detected with Anti-M-24 are components of the muscle cell membrane: (a) these macromolecules are free to diffuse laterally within the myoblast membrane; (b) Anti-M-24, in the presence of complement, induced lysis of the muscle cell membrane; and (c) intact monolayers of viable myoblasts completely absorbed reactivity of Anti-M-24 for myoblasts . These antigens are not loosely adsorbed culture medium components or an artifact of tissue culture because: (a) absorption of Anti-M-24 with homogenized embryonic muscle removed all antibodies to cultured myoblasts; (b) Anti-M-24 reacted with myoblast surfaces in vivo; and (c) absorption of Anti-M-24 with culture media did not affect the titer of this antiserum for myoblasts . We conclude that myogenic cells at all stages of development possess externally exposed antigens which are undetected on other embryonic and adult chick tissues . In addition, myoblasts exhibit surface antigenic determinants that are either masked, absent, or present in very low concentrations on skeletal muscle fibroblasts, embryonic myotubes, or adult myofibers . These antigens are free to diffuse laterally within the myoblast membrane and may be modulated in response to appropriate environmental cues during myodifferentiation.

J Cell Physiol, 1979 Mar, 98(3), 597 - 601
Growth of human embryonic fibroblasts at clonal density: concordance with results from mass cultures; Smith JR et al.; In the past, it has been difficult to grow human diploid fibroblast cells at clonal densities . Newly devised cell culture media and rigorously controlled environmental conditions have greatly increased the ease with which such cells can be cloned . The present work was undertaken to determine whether, under appropriate conditions, diploid fibroblast cells from human embryonic lung, grow as well at clonal densities as in mass culture . The parameters studied were: (1) population doubling time, (2) in vitro proliferative capacity, (3) attachment, (4) percentage of non-dividing cells . In all cases essentially the same results were obtained for cultures at clonal densities and mass cultures . These results indicate that the behavior of these types of cells in clonal culture can be used to infer the behavior of individual cells and clones within a mass culture.

Res Vet Sci, 1979 Mar, 26(2), 259 - 60
Attempts to immunize rats against infection with Fasciola hepatica using in vitro culture antigens from newly excysted metacercariae; Davies C et al.; Attempts were made to immunise rats against Fasciola hepatica using the culture products obtained from the in vitro cultivation of newly excysted metacercariae . Three culture regimes were chosen: (1) medium NCTC 135 for 48 h (2) NCTC 135 + 20 per cent fetal calf serum (FCS) for 48 h (3) NCTC 135 + 20 per cent FCS for 14 days . The used culture medium from each of these regimes was concentrated, mixed with adjuvant and injected subcutaneously into rats . Similarly treated unused culture media was used in control rats . The rats were challenged with an oral dose of 20 F hepatica metacercariae 35 days later and autopsied 96 days after the start of the experiment . The fluke burdens in those rats which had received the culture antigens did not differ significantly from those in the control groups.

Mycopathologia, 1979 Feb 28, 66(3), 161 - 7
Inhibitory effects of condiments and herbal drugs on the growth and toxin production of toxigenic fungi; Hitokoto H et al.; The effects of thirteen kinds of powdered herbal drugs and seven kinds of commercial dry condiments on the growth and toxin production of Aspergillus parasiticus, A . flavus, A . ochraceus, and A . versicolor were observed by introducing these substances into culture media for mycotoxin production . Of the twenty samples tested, cinnamon bark completely inhibited the fungal growth, while the others only inhibited the toxin production . The inhibitors were easily extracted from the samples with solvents such as hot water, chloroform, or ethanol . The extracts from coptis, philodendron bark, mustard, green tea leaves, and zanthoxylum completely inhibited the aflatoxin production of A . parasiticus, however, they had little or no inhibitory effect against A . flavus.

J Cell Biol, 1979 Feb, 80(2), 492 - 8
Fibroblast cellular and plasma fibronectins are similar but not identical; Yamada KM et al.; Fibronectins are multimeric, adhesive glycoproteins present on cell surfaces and circulating in blood . Cellular fibronectin produced by fibroblasts in vitro and fibronectin isolated from plasma are known to be very similar immunologically and biochemically . We investigated whether or not they are identifical . Purified chicken and human cell-surface fibronectins are 150-fold more active in hemagglutination of fixed erythrocytes than plasma fibronectins . Cell-surface fibronectin is also 50-fold more active in restoring a more normal morphology to transformed cells originally missing the protein . However, in two other assays that measure cell attachment to collagen and cell spreading, cell-surface and plasma fibronectins have identical specific activities . In sodium dodecyl sulfate polyacrylamide gels, the subunits of human and chicken plasma fibronectins have significantly smaller apparent subunit molecular weights than cellular fibronectins present on cell surfaces or secreted into culture media . These differences are also present in a characteristic large subfragment of both forms of fibronectin after limited proteolysis by trypsin . We conclude that by both biological and biochemical criteria, cellular and plasma fibronectins are similar but not identical.

Gann, 1979 Feb, 70(1), 21 - 8
An attempt for improving plating efficiency of human diploid fibroblasts; Namba M et al.; An attempt was made to enhance the plating efficiency of normal human diploid skin fibroblasts by testing various culture media . Eagle's minimum essential medium (MEM), Dulbecco's MEM, William's medium E, DM-160, McCoy's 5a, RPMI-1640, Ham's F12, MCDB 102, Waymouth's MB 752/1, and L-15 synthetic media were examined . L-15 was the best among them . Plating efficiency with L-15 supplemented with 20% calf serum ranged from 20 to 60% . Plating efficiency decreased in the order of Eagle's MEM, Dulbecco's MEM, DM-160, and Ham's F12 . Waymouth's MB 752/1, MCDB 102 McCoy's 5a, RPMI-1640, and William's medium E were not favorable for high plating efficiency . Although L-15 gave the best plating efficiency and Waymouth's MB 752/1 the worst, the Waymouth medium supported better cell proliferation than L-15, when cell proliferation was compared in mass culture with these two media . It was concluded that L-15 was better suited for culture of small number of cells, and especially for work involving colony formation.

J Chromatogr, 1979 Jan 21, 168(2), 385 - 93
Separation of unconjugated pteridines by high-pressure cation-exchange liquid chromatography; Stea B et al.; In the course of determining the levels of unconjugated pteridines occurring in various biological fluids, such as urines, plasma and tissue culture media, a method has been developed for the separation and quantitative determination in the picomole range of ten 2-amino-4-hydroxy substituted pteridines . This method involves separation by high-pressure cation-exchange liquid chromatography and fluorescence detection of the eluted compounds at 450 nm . Optimal separation was obtained by isocratic elution with 3 mM phosphoric acid-7% methanol-1% acetonitrile at a flow-rate of 2 ml/min or with 1 mM ammonium dihydrogenphosphate pH 2.8-7% methanol-5% acetonitrile at a flow-rate of 1.5 ml/min . With either solvent, the order of elution of the compounds is: isoxanthopterin, pterin-6-carboxylic acid, xanthopterin, pterin-6-carboxaldehyde, D-erythro-neopterin, L-threo-neopterin, biopterin, 6-hydroxymethylpterin, pterin, 6-methylpterin . In addition, a systemic investigation of the effects of ammonium ion concentration and pH of the solvent as well as column temperature on the separation of these compounds was also conducted.

J Biol Chem, 1979 Jan 10, 254(1), 224 - 32
Separate amino and carboxyl procollagen peptidases in chick embryo tendon; Leung MK et al.; Procollagen synthesized by freshly excised chick enbryo leg tendons is efficiently processed by proteolytic removal of first the amino propeptides and then the carboxyl propeptides . The same processes proceed in confluent short term cell cultures derived from such tendon explants; in sparse cultures cleavage of the amino propeptides predominates . Separate amino and carboxyl procollagen peptidase activities were demonstrated by specific assays in enzymes obtained from cell culture media by ammonium sulfate precipitation, ion exchange chromatography, and velocity sedimentation . Both enzymes are inhibited by EDTA and 1:10 phenanthroline but not by inhibitors of serine proteases . Evidence is provided that the proteolytic scissions are specific and similar to the physiologically occurring processes . The collagen telopeptides left after cutting by the enzymes can participate in lysyl oxidase-induced cross-linking . The enzymes can remove propeptides from cross-linked procollagens without destroying these links which occur through telopeptides . The enzymes act on the separated amino and carboxyl portions of procollagen fragmented by vertebrate collagenase and can act on procollagens which have been associated as well as on molecules in solution.

J Supramol Struct, 1979, 11(2), 217 - 25
Human fibrosarcoma cells produce fibronectin-releasing peptides; Keski-Oja J et al.; A sensitive radioimmunoassay, specific for human fibronectin, was used to measure the ability of certain biologically active polypeptides to release fibronectin from cultured human lung fibroblasts into their culture media . Concentrated, serum-free supernatant from a human fibrosarcoma cell line was fractionated by gel filtration chromatography in the presence of acetic acid . Various polypeptides with molecular weights between 46,000 and 6,000 were tested for their ability to release fibronectin from cells . The column fraction, containing polypeptides with an apparent molecular weight of 10,000, exhibited the ability to rapidly release fibronectin from target cells . The activity could be inhibited by phenylmethyl sulphonylfluoride . Several other hormonal factors, tested in parallel with the column fractions, failed to show this effect . The 10,000 dalton molecular weight polypeptides may represent a family of cellular gene products responsible for maintenance of low levels of surface associated fibronectin in fibrosarcoma cells and thus be related to their infiltrating properties by preventing the formation of the extracellular matrix.

Arch Exp Veterinarmed, 1979, 33(6), 957 - 9
{Experience regarding the suitability of liquid and solid PPLO culture media produced in GDR for mycoplasma cultivation}; Templin G; Proper isolation of mycoplasma from milk or tissue was found to entail exacting demands on nutrient substrates . Several institutes of the GDR have jointly developed and tested a medium which consists exclusively of GDR-made substances . It is a culturing medium which is suitable for detection and identification of acholeplasma and mycoplasma species in milk and organ samples taken from cattle.

J Biol Stand, 1979, 7(4), 275 - 84
A method for testing the growth activity of tissue culture media using suspension lymphoblastoid cells; Babkova H et al.; A new rapid method for testing the growth activity of tissue culture media using the 'Raji' and 'Simpson' strains of lymphoblastoid cells grown in suspension is described . This method has advantages for rapidly assessing the quality of a culture medium.

Exp Cell Biol, 1979, 47(5), 332 - 50
Enzymatic modulation of the cell surface in malignant transformation of normal human mammary epithelial cells and in conversion of mammary carcinoma cells; Hakim AA; The effect of treatment with trypsin and neuraminidase on morphology, proliferation pattern, adhesiveness and ultrastructure of human normal mammary and carcinoma cell lines has been investigated . These criteria were chosen because they reflect on the cell surface function . Active enzymes were required to produce changes . If trypsinized and then allowed to grow in culture media supplemented with 'fibroblast growth-promoting factor', human normal mammary epithelial cells proliferated in a disorganized pattern with cells overlapping and piling up and developing ultrastructural characteristics of neoplastic cells . On the other hand, if treated with neuraminidase and then permitted to grow in presence of the 'fibroblast growth-promoting factor', mammary carcinoma cells proliferated in an organized pattern, forming one-cell-thick, fibroblast-like monolayers and ultrastructural characteristics of normal epithelial cells.

Adv Exp Med Biol, 1979, 112, 283 - 91
Action of porcine follicular fluid oocyte maturation inhibitor in vitro: possible role of the cumulus cells; Hillensjo T et al.; To study the mode of action of porcine follicular fluid oocyte maturation inhibitor isolated cumulus-enclosed or mechanically denuded pig oocytes were used . Two types of culture media were employed, a complex containing 15% pig serum (TC199A) and a defined, minimal medium (BMOC) . The maturation of cumulus-enclosed oocytes was comparable in the two types of culture media, but only in the complex medium did the cumulus cells remain functional in terms of morphology and progesterone secretion . The low molecular weight portion of follicular fluid partially inhibited oocyte meiosis and cumulus progesterone secretion in 199A . No inhibition of oocyte maturation was seen when follicular fluid was added to BMOC . Since denuded oocytes did not respond to follicular fluid in either culture medium, it is suggested that the cumulus cells may mediate the action of the follicular fluid oocyte maturation inhibitor.

Endocrinology, 1979 Jan, 104(1), 97 - 100
Evidence for the autoregulation of hormone secretion by prolactin; Herbert DC et al.; The cells from the 2B8 clonal strain of pituitary cells which previously have been reported to produce only PRL were incubated with varying amounts of ovine PRL (oPRL) ranging from 0.01--1000 ng/ml . After 1 h, significantly less PRL was measured in culture media from cells incubated with all doses of oPRL employed . This inhibition was dose related . The cells were rinsed five times with Hank's balanced salt solution and subsequently placed into PRL-free media for an additional hour . Hypersecretion of hormone occurred from those cells previously exposed to 1--1000 ng/ml oPRL . The PRL in the media from cells previously exposed to 0.01--0.1 ng/ml was similar to that in the control culture . In some experiments, the cellular concentration of PRL was determined at the end of each of the two 1-h incubation periods . After the initial exposure to oPRL, an increase in intracellular hormone was observed only in those groups incubated with 1--1000 ng/ml oPRL . In contrast, after the cells were rinsed and placed into PRL-free medium, the cellular concentration of PRL was unchanged in all groups except the group previously exposed to 100 ng/ml oPRL . These data indicate that PRL can inhibit its own secretion through direct action on the lactotrophs of the anterior pituitary gland.

Endocrinology, 1979 Jan, 104(1), 198 - 204
Autonomous secretion of follicle-stimulating hormone by long term organ cultures of rat pituitaries; Sheridan R et al.; Pituitaries removed from ovariectomized adult rats were maintained for 18 weeks in organ culture using three different culture media . Gonadotropin secretion was assessed by RIA and was correlated with the histological features of the cultures . In medium favoring prolonged survival of the cultures, LH content of the medium fell to a low level within a few days . In the same cultures, FSH production initially decreased before increasing and leveling at a plateau which persisted until the end of the culture period . Cultures in medium unsuitable for long term survival of pituitary tissue displayed a similar decrease in LH production along with a gradual fall of FSH . It was concluded that contrary to LH, FSH may be secreted autonomously by pituitaries removed from hypothalamic control, provided that culture conditions are adequate for survival of gonadotropes.

Somatic Cell Genet, 1979 Jan, 5(1), 23 - 8
Cloning of Drosophila cells: effect of vitamins and yeast extract components; Wyss C; Yeast extract, a component of Drosophila cell culture media, is shown to contain substances of high, intermediate, and low molecular weight, that are, respectively, essential, inhibitory, and stimulatory for colony formation in semisolid agar medium . Furthermore, it is shown that high concentrations of pyridoxal greatly increase the cloning efficiency of Drosophilia cells . A cloning method with line Kc is described which routinely gives cloning efficiencies in excess of 20%.

Can J Microbiol, 1979 Jan, 25(1), 53 - 60
{Identification of an extracellular nucleotide pyrophosphatase in the culture media of Streptomyces mediterranei ME/R 17}; Pellon G et al.; An exocellular pyrophosphatase, active on the nucleotide precursors of peptidoglycans, has been found in the culture medium of Streptomyces mediterranei ME/R 17 . This enzyme was separated from the DD-carboxypeptidase by batchwise adsorption on DEAE cellulose . The pyrophosphatase had no strict substrate requirements, it hydrolyzed various UDP-sugar substrates: UDP-GlcNAc, UDP-Mur NAc and UDP-MurNAc peptides, giving rise to the corresponding sugar phosphate and to UMP . The enzyme preparation also contained a 5'-nucleotidase activity and UMP was further split to give uridine . This nucleotidase activity was inhibited by potassium tetraborate . Both cytoplasmic and particulate preparations from cells of S . mediterranei also contained a pyrophosphatase activity while only the particulate fractions showed the DD-carboxypeptidase activity . The pyrophosphatase excretion was tested during the grwoth cycle . The activity of the enzyme showed a constant increase throughout the exponential growth and a stronger increase in the late exponential phase . Such a result could be correlated with a consumption of the nutrients in the culture medium, in fact a relatively poor culture medium had a strong positive effect upon the production of the exocellular pyrophosphatase.

Pharmacology, 1979, 18(2), 95 - 102
Relationship between surface activity and toxicity to Chang liver cultures of tricyclic antidepressants; Yasuhara H et al.; Chang liver cell cultures were exposed to the tricyclic antidepressants, chlorimipramine (CIM), nortriptyline (NT), amitriptyline (AT), imipramine (IM), and dosepin (DOX) . Loss of enzymes into surrounding media and cytopathic changes were used to quantitate cytotoxicity . Time- and concentration-related cytotoxic effects were evident for all drugs . The order of cytotoxic potency was CIM greater than NT greater than AT greater than IM greater than DOX . All tricyclic antidepressants tested lowered the surface tension of the salt solution contained in the tissue culture media and the order of their surface activity was identical to that of their cytotoxicity . It is postulated that the cellular toxicity induced by tricyclic antidepressants in vitro is related to a function of their surface activity.

Ann Ist Super Sanita, 1979, 15(1), 77 - 84
{Methods for collection, transport and culture media}; Ercolani F et al.; A crucial factor affecting the ultimate success of anaerobic cultures is a proper specimen collection, with care to avoid inclusion of normal flora often present on human mucosa . In general, material for anaerobic cultures is best obtained using a needle and syringe, from which the air must be expelled . Then the specimens should be placed immediately into an anaerobic transport broth and then in suitable culture pre-reduced media.

J Hyg Epidemiol Microbiol Immunol, 1979, 23(4), 397 - 406
Influence of culture medium of the fatty-acid profile in enteric bacteria; Vasyurenko ZP et al.; Enteric bacteria having a high content of cyclopropane fatty acids steeply increase their synthesis when grown on insufficiently propitious culture media (meat-peptone agar or modified Drobot'ko synthetic medium) as compared with bacteria grown under more favourable conditions (meat-peptone broth) . Simultaneously, a decrease in monounsaturated fatty acids and increase in palmitic acid are observed . One of the main factors underlying the change in the proportion of fatty acids in bacteria grown on synthetic medium is an increase in medium pH in the process of their growth . Enteric bacteria containing minute amounts/or not containing cyclopropane fatty acids at all (under the experimental conditions used) change their fatty-acid profile little if the culture medium is changed . When grown under insufficiently favourable conditions, these bacteria mainly display an enhanced content of palmitic acid and a lowered content of octadacenoic acid as compared with bacteria grown under more favourable conditions . Of the culture media used, meat-peptone broth, which affords the most favourable conditions for eneteric bacteria growth, is the most suitable medium for obtaining data of taxonomic value.

Arch Exp Veterinarmed, 1979, 33(3), 419 - 28
{Studies of bovine Mycoplasma mastitis . 2 . Testing of various culture media and culture methods for the isolation of Mycoplasma from milk samples}; Pfutzner H et al.; The recipes Medium-I broth, Medium-B broth, and Weissenseemycoplasma broth as well as Medium-I agar, Medium-B agar, and MRL agar were tested for their applicability to culturing mycoplasma from milk samples, using the direct and indirect techniques . No dependable information on the occurrence of mycoplasma was obtainable from tested material unless several nutritive media and techniques were combined . Medium I, for which almost no imported substances were needed, was in no way inferior to common international nutritive substrates . Its use in conjunction with the indirect culturing technique is recommended for routine diagnosis.

Am J Vet Res, 1979 Jan, 40(1), 130 - 4
Evaluation of various methods for the detection of enteropathogenic Escherichia coli in diarrheic calves; Lariviere S et al.; Evaluation of the Escherichia coli population in the small intestine of diarrheic calves was performed by estimating the number of colony-forming units and the observation of direct gram-stained smears of mucosal scrapings . Enterotoxigenicity of the isolates was determined by the infant mouse test and the ligated intestinal segment in calves . The influence of various broth culture media on the production of enterotoxin was also studied . Serologic characterization of the E coli isolates was performed . A total of 190 cases of diarrhea in bovine neonates was studied and it was found that enteropathogenic E coi was not responsible for more than 20% of the cases . A practical approach for the recognition of enteropathogenic E coli in a routine diagnostic laboratory is proposed.

Int Arch Allergy Appl Immunol, 1979, 60(4), 407 - 13
Mitotic activity of thymocytes in a synthetic tissue culture medium . Effect of L-alanine; Sandberg G et al.; DNA synthesis in guinea pig thymocytes suspended in RPMI 1640 medium increased to a peak after 4-5 h in culture and was followed by increased mitotic activity, indicating that many thymocytes in S phase proceeded through G2 into mitosis . Addition of L-alanine to the medium markedly increased the DNA synthesis within 1 h and the mitotic frequency from 6 h . The increase in DNA synthesis when L-alanine was present in the medium was thus caused by an increased number of cells in S phase . Human thymocytes cultured in RPMI 1640 for 18 h had a low mitotic frequency . Addition of L-alanine immediately started DNA synthesis in the arrested thymocytes resulting in increased mitotic activity from 6 h later . The results show that L-alanine is a growth factor for guinea pig and human thymocytes and should be included in tissue culture media used for such cells . Growth of thymocytes in vitro was partly synchronized, and the mitotic studies indicated that many cells had entered S phase near the start of incubation.

Oncology, 1979, 36(5), 224 - 7
Increased colony-stimulating activity in the plasma of a patient with lymphoepithelioma metastatic to the liver; Tebbi K et al.; Leukocytosis with predominance of poly morphonuclear leukocytes is not unusual in a variety of primary or metastatic hepatic tumors . However, to date no apparent explanation for this event is available . This paper describes extreme leukocytosis in a 17-year-old black male with lymphoepithelioma after development of metastases to the liver . In order to identify the source of leukocytosis, studies of granulocytic colony-stimulating activity (CSA) of the patient's plasma and leukocyte-conditioned media (LCM) were carried out and compared with normal controls . The assay system consisted of nonadherent marrow cells of hematologically normal individuals and normal mice cultured in semi-solid culture media . In this system, the number of granulocytic colony-forming units (CFU-C) is proportional to the amount of stimulating activity present, and therefore allows for quantitative comparison . The number of colonies produced under the influence of patient's plasma was significantly higher (p less than 0.001) than those obtained from the patient's LCM or from control plasma or LCM . This pattern is the reverse of that seen in normal individuals in whom the major sources of CSA are macrophages and monocytes . It is conceivable that malignant cells are the source of additional CSA.

Rev Chir Orthop Reparatrice Appar Mot, 1979 Jan-Feb, 65(1), 39 - 43
{Mycobacterium fortuitum infection after total hip prosthesis . A report of 3 cases (author's transl)}; Badelon O et al.; The authors have observed three instances of sepsis due to Mycobacterium fortuitum complicating total hip replacement for osteoarthritis . The case histories are fully described . The requirements are given for recognition of the organism which grows on special culture media and which may be mistaken for Mycobacterium tuberculosis . This feature explains why, in some cases, the organism may not be discovered . Antibiotics were ineffective but the general condition of the patient was not greatly affected.

Rev Chir Orthop Reparatrice Appar Mot, 1979 Jan-Feb, 65(1), 39 - 43
{Mycobacterium fortuitum infection after total hip prosthesis . A report of 3 cases (author's transl)}; Badelon O et al.; The authors have observed three instances of sepsis due to Mycobacterium fortuitum complicating total hip replacement for osteoarthritis . The case histories are fully described . The requirements are given for recognition of the organism which grows on special culture media and which may be mistaken for Mycobacterium tuberculosis . This feature explains why, in some cases, the organism may not be discovered . Antibiotics were ineffective but the general condition of the patient was not greatly affected.

Zentralbl Bakteriol Naturwiss, 1979, 134(6), 547 - 50
Production and isolation of milk-clotting enzyme from Aspergillus versicolor; Abdel-Fattah AF et al.; The production of a milk-clotting enzyme by Aspergillus versicolor in 19 different culture media was investigated . Considerable milk-clotting activity was achieved by supplying corn steep liquor with either glucose of maltose . Dephytinization of corn steep liquor had an adverse effect on the production of milk-clotting enzyme . The results indicated that complex organic compounds favoured the production of the enzyme . Precipitation with acetone or tannin was unsuitable, but ammonium sulphate and ethanol above certain concentration produced active fractions.

Acta Haematol, 1979, 61(3), 138 - 43
Effect of hemin and Protoporphyrin IX on the protein-synthesizing activity of human granulocytes, lymphocytes and platelets; Malik Z et al.; The hemin effect on protein synthesis of human granulocytes, lymphocytes and platelets was examined . Hemin added to culture media without serum caused a dose-dependent inhibition of protein synthesis in all three cell types . A cell-specific enhancement of protein-synthesizing capability was observed in 24-hour cultures in the presence of hemin and serum . A marked increase of protein synthesis was found in granulocytes, unchanged in lymphocytes and decreased in platelets . Lymphocytes from patients with chronic lymphatic leukemia (CLL) were moderately inhibited by hemin when incubated in media containing serum, the effect being more pronounced in the presence of freshly disolved doses of hemin . Addition of protoporphyrin IX to cells cultures resulted in a marked suppression of protein synthesis by the three cell types, in all experiments . These results confirm the importance of serum proteins in preventing the inhibitory effects of free hemin and protoporphyrin IX on blood cell protein synthesis . On the other hand, they show a cell-specific enhancement of the protein-synthesizing capacity mediated by hemin.

Zentralbl Bakteriol Naturwiss, 1979, 134(7), 594 - 9
Effect of ruthenium on nitrogen fixation by some nitrogen fixers; Bahadur K et al.; The effect of ruthenium chloride in the culture media on the nitrogen-fixing ability of the three nitrogen fixers (unidentified species of Azotobacter, designated here as D3, B3, and B), isolated from Allahabad soil, was studied . It was observed that the nitrogen-fixing ability of the organisms is much increased in presence of 25-75 micro M concentration of ruthenium chloride in the culture media, while sugar consumption remains more or less steady . Also, if mg nitrogen fixed/g carbon consumed in two culture media with successive increasing concentrations of ruthenium chloride is compared by calculating the difference in increase of the amount of nitrogen fixed and carbon consumed in these two culture media, it was observed that high amounts of nitrogen are fixed by B3 and B between 25-50 micro M, and between 50-75 micro M concentration of ruthenium chloride by D3.

Oncology, 1979, 36(2), 72 - 5
Unexpectedly low incorporation of isotopic acetate into lipids of Ehrlich ascites tumor cells cultured in lipid-poor medium; Loffler M et al.; The present communication reports data on the lipid biosynthesis of Ehrlich ascites tumor cells grown in culture media supplemented with modified sera . Whereas the metabolisms of {14C}pyruvate and {14C}mevalonate are identical in all media tested, the incorporation of {14C} acetate is higher in medium with dialyzed serum than in medium with delipidized serum; it is suppressed in the absence of all lipids in culture medium . Cellular integrity is not impaired in modified media . The results indicate that acetyl-CoA synthetase of Ehrlich ascites tumor cells is not regulated by exogenous lipids as is known to be the case in nonmalignant cells.

Prostaglandins Med, 1978 Dec, 1(6), 433 - 9
Adriamycin stimulates canine kidney (MDCK) cells to deacylate cellular lipids and to produce prostaglandins; Ohuchi K et al.; Dog kidney (MDCK) cells treated with adriamycin (0.5 micrograms/ml) for 1 hr, produced from 2 to 7 times more prostaglandins E2 and F2alpha when measured in culture media 24, 48 and 72 hrs after the treatment . Indomethacin (ID50 less than 2 x 10(-8) M) and cycloheximide (0.5 micrograms/ml) inhibited this adriamycin-stimulated prostaglandin production . The aglycone of adriamycin (0.5 to 5.0 micrograms/ml) had little stimulating effect . Treatment of {3H}arachidonic acid-labeled MDCK cells with adriamycin (0.5 micrograms/ml) for 1 hr also stimulated deacylation of cellular lipids during subsequent incubation . Altered morphology of MDCK cells resulted from such treatment with adriamycin; indomethacin did not inhibit this change, but cycloheximide did.

Acta Pathol Microbiol Scand {B}, 1978 Dec, 86B(6), 349 - 53
"Runde" virus: further characteristics and a method for purification; Traavik T et al.; Neither BHK 21/c13, BSC-1, Vero nor GMK cells were of use for quantification of "Runde" virus . The titres were low and difficult to reproduce . Infected newborn mouse brains gave considerably higher yields than any of the cell cultures . The growth curve in BHK 21/c13 cells showed a slow increase in both intracellular and extracellular virus until maximum titres of about 10(6) baby mouse LD50 were reached at 48 and 72 hours post-infection (p.i.) . During the following 24 hours, the infectivity dropped by about 1 log10 unit and was then unaltered until 196 hours p.i . Infected BHK 21/c13 cells did not haemadsorb chicken erythrocytes, although the culture media contained haemagglutinins . Resistance to BUdR indicated an RNA genome . Concentrated and purified virus preparations were produced by polyethylene glycol 6000/NaCl "precipitation" and hydroxylapatite column chromatography . Treatment with a colloidal silica gel widened the spectrum of agglutinable erythrocyte species.

Tijdschr Diergeneeskd, 1978 Nov 15, 103(22), 1217 - 30
{Mycoplasma synoviae control . I . Studies on the thermal sensitivity of pathogenic avian mycoplasmas (Mycoplasma synoviae, Mycoplasma gallisepicum and Mycoplasma meleagridis)}; Goren E; A number of experiments were carried out to study the thermal sensitivity of Mycoplasma synoviae and Mycoplasma gallisepticum as well as that of young embryos in vitro and in ovo . A full mycoplasmacidal effect was attained after heating in bouillon cultures for six hours at 45 degrees C (including cultures of Mycoplasma meleagridis), for two and a half hours at 50 degrees C, for ninety minutes at 52 degrees C and for thirty minutes at 55 degrees C . Yoder's method of heating to control these mycoplasmas in hatching eggs was found to be inadequate . When inoculated eggs were heated for ten hours at 45 degrees C, mycoplasmas could no longer be isolated; however, this had a highly adverse effect on the proportion of eggs hatched (33 and 50 percent for hen's eggs and turkey eggs respectively) . Mycoplasma synoviae was slightly more sensitive to heating than Mycoplasma gallisepticum, whereas Mycoplasma meleagridis showed more resistance than Mycoplasma gallisepticum . After heating, atypical colonies constantly appeared on the primary plates and conversion of glucose was delayed in the cultures . Isolation by SPF embryos was found to be a more sensitive method than isolation using artificial culture media only . When fresh incubated eggs were heated for ten hours at 45 degrees C, for two and a half hours at 46 degrees C or for thirty minutes at 47 degrees C, fifty percent of embryos died, whereas one hundred per cent died after heating for sixty minutes at 47 degrees C . Thermal treatment of hatching eggs to eliminate Mycoplasma synoviae, Mycoplasma gallisepticum and Mycoplasma meleagridis is unsuitable for use in poultry practice.

Biochem J, 1978 Nov 15, 176(2), 455 - 62
Studies on the fate of pulmonary surfactant in the lung; Desai R et al.; 1 . Radioactively labelled pulmonary surfactant was prepared in an isolated perfused lung system provided with {14C}hexadecanoate . 2 . After intratracheal administration of pulmonary surfactant radioactively labelled components were rapidly distributed into different lung fractions, including macrophages (free cells), but most of the radioactive label was accumulated by the lung tissue . 3 . Alveolar macrophages, maintained in a variety of culture media in the presence and absence of mineral particles, incorporated a low percentage (11%) of radioactively labelled components when incubated with the surfactant, although evolution of labelled CO2 (6% of the original total activity) suggested that some breakdown of the components had taken place . 4 . In similar cultures little intracellular accumulation or extracellular release of non-esterified fatty acids was demonstrated, indicating minimal catabolism of the high-molecular-weight lipid components of surfactant (particularly phosphatidylcholine) . 5 . However, experiments in vitro designed to simulate the lysosomal degradation of endocytosed surfactant indicated that the macrophage had enzymes capable of releasing non-esterified fatty acids, particularly hexadecanoate, from the lipoprotein complex . 6 . It is argued that lung cells, other than alveolar macrophages, may also have a role in surfactant turnover.

Cell Tissue Res, 1978 Nov 9, 194(1), 55 - 69
Secretion of melanophore-stimulating hormone (MSH) in long-term cultures of pituitary neurointermediate lobes; Semoff S et al.; Neurointermediate lobes from pituitaries of the frog, Rana berlandieri forreri (Rana pipiens, sensu lato), were maintained in organ culture in media with and without serum for up to six months . The cultured tissues were examined periodically by light microscopy and transmission electron microscopy and by bioassay of the melanophore-stimulating hormone (MSH) secreted and present in the culture media . Light-microscopic observations revealed a high degree of preservation of the pars intermedia at four weeks with isolated areas of some glands maintaining histological integrity for the entire six months . Similarly, at the ultrastructural level the cells appeared morphologically intact and to be actively synthesizing and secreting hormone . Bioassays showed the glands to be continuously secreting MSH; however, larger yields of hormone were obtained in media lacking serum . No significant ultrastructural differences between cells grown in the presence or absence of serum were detected . The difference in concentration of MSH between the two groups therefore apparently results from enzymatic degradation of the hormone by the serum . Organ culture of the vertebrate neurointermediate lobe may provide a unique method for the production of large quantities of MSH and for the study of other melanotropic and opiate peptides as they may be synthesized and secreted by the pars intermedia.

J Cell Physiol, 1978 Nov, 97(2), 221 - 9
Differential effects of hydrocortisone on both growth and collagen metabolism of human fibroblasts from normal and keloid tissue; Russell JD et al.; Cultured fibroblasts isolated from normal and keloid tissue do not differ in their growth characteristics or in the rate of collagen synthesis under routine culture conditions . The addition of hydrocortisone to the culture media results in significant differences in both growth and collagen synthesis between these cell types . Collagen synthesis is inhibited 60% in normal cultures by hydrocortisone (0,5 micrograms/ml) and the population size at which density-dependent growth inhibition is achieved is increased . Keloid-derived fibroblasts grow to a lower maximum density in the presence of hydrocortisone, while their rate of collagen synthesis is not significantly reduced . The rate of non-collagen protein synthesis is increased significantly by hydrocortisone in both cell types . Comparison of normal and keloid-derived cultures obtained from a single individual suggests that the keloid phenotype with respect to both growth and collagen synthesis is restricted to the fibroblasts isolated from the keloid nodule.

J Biochem (Tokyo), 1978 Nov, 84(5), 1171 - 6
Collagenolytic activities of cultured human malignant melanoma cells; Tane N et al.; Three human malignant melanomas were cultured in pure populations and one tumor was cloned into melanotic and amelanotic cell lines . In the homogenates of these cultured cells, specific collagenase activities were demonstrated by isotope release from 14C-labeled collagen, disc electrophoresis, and specific cleavage of collagen molecules as demonstrated in the segment long spacing form . No significant collagenase activity was observed in the culture media . Interestingly, early cultures had a high collagenase activity in the cells and as they were successively subcultured, the activity diminished . Cysteine completely inhibited the degradation of tropocollagen as determined by disc electrophoresis and EDTA partially inhibited the degradation . It is concluded that human malignant melanoma cells produce a specific collagenase in vitro which can be extracted in early culture directly from the homogenate.

Schweiz Med Wochenschr . 1978 Oct 14;108(41):1607.
{Isolation of human tumor cells: conditions for in vitro diagnostic and therapeutic studies (proceedings)}; Peter HJ et al.; With a view to the possible use of human tumor cells for diagnostic and therapeutic tests (e.g . determination of hormone receptors, sensitivity to cytotoxic drugs, cytotoxicity tests), an attempt has been made to isolate and cultivate pure human tumor cell populations . 49 malignant effusions from 42 patients formed the starting material . The tumor cell isolation was carried out in three steps: 1 . elimination of erythrocytes by density-gradient centrifugation or osmotic lysis with triple distilled water; 2 . concentration of tumor cells by lg-sedimentation; 3 . elimination of adherent cells by short term cultivation in plastic tissue culture flasks . This procedure yielded 95-98% tumor cells . From these suspensions of purified tumor cells, primary cultures were successfully established by supplementing the tissue culture media with autologous effusion fluid.

J Neurol Sci, 1978 Oct, 38(3), 409 - 19
Globoid cell leukodystrophy (Krabbe's disease) . Metabolic studies with cultured fibroblasts; Tanaka H et al.; Metabolism of tritium-labelled galactosylceramide and lactosylceramide added to the culture medium was examined in cultured skin fibroblasts from 4 patients with globoid cell leukodystrophy (GLD) and 4 control individuals . The uptake of {3H}galactosylceramide and {3H}lactosylceramide by the fibroblasts continued actively at least up to 3 days . Approximately 30--40% of the galactosylceramide, which had been taken up, was released subsequently from the cells in a 4-day period, whereas only 10% of lactosylceramide was released during the same period . The GLD fibroblasts showed no abnormality in the kinetics of the uptake and in the release of these glycosphingolipids which are natural substrates of the beta-galactosidase genetically deficient in the disorder . This finding differs from that reported for fibroblasts from patients with metachromatic leukodystrophy, which showed abnormal accumulation and retention of sulfatide added to the culture media . However, degradation of added galactosylceramide to {3H}galactose by the GLD fibroblasts was only 25% of the control cells, while lactosylceramide was degraded at 70% of the normal rate . These findings are consistent with the known substrate specificities of the two acidic beta-galactosidases in human tissues; galactosylceramide is hydrolyzed almost exclusively by galactosylceramidase, while lactosylceramide can be hydrolyzed by both galactosylceramidase and GM1-ganglioside beta-galactosidase.

Proc Natl Acad Sci U S A, 1978 Oct, 75(10), 5226 - 9
Growth cone formation in cultures of sensory neurons; Bray D et al.; Three experimental situations have been found in which cultured sensory neurons from embryonic chicken will form growth cones from positions along the length of the neurite . If the neurons are dissected with a remaining short axonal stump and plated into serum-free medium, they can form a morphologically normal growth cone from the stump within 15 min, even in the presence of cycloheximide or puromycin . When neurites growing in culture media with low levels of serum are cut at any point with microneedles, growth cones are produced quickly from the amputated stump, usually within 20 min . Treatment of growing neurons with low concentrations of colchicine, Colcemid, or podophyllotoxin results in the progressive appearance of lateral filopodia and regions of flattened cytoplasm that closely resemble growth cones except for their preterminal positions . These observations show that the potential to form growth cones is distributed throughout the neuron and suggest that this normally repressed in some way by the neuronal microtubules.

Lab Anim Sci, 1978 Oct, 28(5), 567 - 74
Comparative mitogen responses of lymphocytes from Colombian Panamanian, and Peruvian owl monkeys; Taylor DW et al.; A comparative study of lymphocyte responses to various mitogens, an index of cell-mediated immune competency, was made under various culture conditions using peripheral blood lymphocytes from Colombian, Panamanian, and Peruvian Aotus monkeys . Dose response curves were determined for each primate group to phytohemagglutinin, concanavalin A, and pokeweed mitogen stimulation . Considerable variation in mitogen response was observed . The influence of culture media supplemented with fetal calf serum or autologous plasma, the number of cells per culture, and the duration of incubation on mitogen responsiveness was also evaluated . Optimal lymphocyte culture conditions also differed among the three groups . Owl monkeys from the same location in South and Central America showed similar responses to physical condition and free of detectable disease, these differences probably reflect fundamental inherent biological differences between subpopulations of owl monkeys.

J Neurocytol, 1978 Oct, 7(5), 623 - 36
The role of calcium ions in the synthesis and transport of noradrenaline carrier vesicles in guinea-pig sympathetic neurons in vitro; Anderson PN et al.; Guinea-pig inferior mesenteric ganglia (IMG)/hypogastric nerve preparations were incubated in tissue culture media containing ethylene glycol-bis-(beta-aminoethyl ether)N,N'tetra-acetic acid (EGTA) or colchicine and examined either by fluorescence microscopy or by transmission electron microscopy . Two millimolar EGTA inhibited the accumulation of noradrenaline (NA) fluophore proximal to a crush, but did not produce an increase in the fluorescent intensity of, or the number of dense-cored vesicles (DCVs) within the neuronal perikarya . Calcium ions, but not magnesium ions, were able to block this effect of EGTA . Preparations incubated in the presence of colchicine (2.5 microgram/ml) showed a reduction in the amount of fluorescent material accumulating proximal to a crush, but an increase in both the fluorescent intensity and the number of DCVs within the neuronal perikarya . The suggestion that calcium ions are required for the synthesis of some part of the NA-containing vesicle rather than for their loading onto the axoplasmic transport mechanism is discussed.

J Bacteriol, 1978 Sep, 135(3), 818 - 27
Mycoplasma growth factors in bovine serum fraction; Washburn LR et al.; Mycoplasma growth factors in bovine serum fraction were separated by Sephadex G150 column chromatography and density ultracentrifugation . The major growth factor of bovine serum fraction eluted from the Sephadex column in the void volume . Its growth-supporting activity was greatly enhanced by the presence of bovine serum albumin in the mycoplasma culture media . Other investigators had previously identified the major growth factor in serum as an alpha-lipoprotein . Although density ultracentrifugation revealed the presence of traces of a high-density lipoprotein in bovine serum fraction, another, less dense component, isolated by ultracentrifugation (component 3) and containing cholesterol, cholesteryl esters, free fatty acids, triglycerides, and protein, but no lipoprotein, exhibited considerably more growth-supporting activity than did the high-density lipoprotein, thus indicating that at least two mycoplasma species do not require intact serum lipoprotein for growth . Both the high-density lipoprotein and component 3 exhibited maximum activity only in the presence of bovine serum albumin . A chloroform extract containing component 3 lipids combined with bovine serum albumin to form an effective, partially defined, less complex substitute for serum in mycoplasma culture media.

Immunology, 1978 Sep, 35(3), 539 - 48
Use of leucocyte migration under agarose to study spontaneous and directed locomotion of leucocytes; Repo H et al.; Three different cell attractants, together with the parallel use of the leucocyte migration agarose test (LMAT) and the leading front modification (LFM) of the Boyden chamber technique, were employed in studying whether the maximal migration of normal human polymorphonuclear leucocytes (PMNs) is higher toward an attractant (chemotaxis) than in the same attractant incorporated in the culture media (chemokinesis) . Using LMAT, the maximal migration distance toward zymosan activated serum (ZAS) was found to be significantly longer than that under agarose mixed with ZAS, thus indicating a chemotactic effect exerted by ZAS . When bacterial culture filtrate (BCF) and casein were used as attractants, the corresponding difference was not significant, implying that the stimulatory effect of these substances on cell migration could be explained by increased random locomotion (chemokinesis) alone . In LFM, the migration rate was significantly higher along a casein gradient than without a gradient . Using ZAS, however, only chemokinesis could be demonstrated . BCF was found to attract PMNs into membrane filters only in the presence of human serum albumin . These observations give credence to the view that both LMAT and LFM are applicable to the in vitro assessment of chemotaxis and chemokinesis but the attractant of choice for this is different in each of the two methods.

Am J Anat, 1978 Sep, 153(1), 81 - 95
Specific subclones derived from a multipotential clone of rat anterior pituitary cells; Shiino M et al.; Several functional subclones of rat anterior pituitary cells were established from our 2A8 clone which apparently contains a heterogenous population of committed and uncommitted cells . On the basis of the hormones secreted into the culture media, as measured by radioimmunoassay, these subclones were divided into four categories, i.e., subclones which secrete (1) ACTH only, (2) prolactin only, (3) prolactin and GH or (4) ACTH, prolactin and GH . None of the subclones produced detectable amounts of thyrotrophic or gonadotrophic hormones . Subclones which secrete a single hormone have shown no change in the type of hormone produced, indicating that these subclones were each derived from a committed cell . The cells of all subclones exhibit a normal diploid karyotype and show good growth characteristics . The cells of the different subclones can be classified by phase contrast microscopy into four categories . However, no clear-cut ultrastructural features have been observed which can be correlated with the different categories of subclones . On the basis of the results a hypothesis is proposed relative to the functional cytodifferentiation of anterior pituitary cells.

Rev Asoc Argent Microbiol, 1978 Sep-Dec, 10(3), 83 - 93
{Influence of the sterilization technic and culture media components on the growth and dissociation of Brucella abortus strain 19 in submerged cultures}; Yantorno OY et al.; Submerged cultures of Brucella abortus strain 19 were studied in shaking flasks . The influence of the sterilization methods and the medium composition on the bacterial yield and cellular dissociation were studied . The selected medium was as follows (amounts in g/l): casein pancreatic hydrolizated 30; yeast extract 10; glucose, 30; sodium phosphate dibasic anhydrous 3,3; sodium monobasic monohydrate 9 . Cell concentration of 8 . 10(10) viable cell/ml was obtained after 48 hours when the medium components were separated and sterilized at 121 degrees C for 20 min in autoclave.

J Allergy Clin Immunol, 1978 Sep, 62(3), 137 - 42
Enhancement of antigen-induced leukocyte histamine release by a mononuclear cell--derived factor; Bamzai AK et al.; Supernatant fluids of phytohemagglutinin-stimulated mononuclear cells from ragweed-sensitive patients significantly enhanced the release of histamine from antigen-triggered leukocytes of ragweed-sensitive as well as control individuals . Supernatants of mononuclear cells from control individuals did not reveal this enhancing effect, nor was it found with the use of supernatants of unstimulated mononuclear cells of ragweed-sensitive patients or culture media with PHA alone . Supernatant fluids of phytohemagglutinin-stimulated mononuclear cells of patients sensitive to trees and grass also revealed this enhancing effect . The factor(s) responsible for the enhancement of antigen-induced histamine is heat labile and has a molecular weight of less than 10,000 daltons . The mechanism and site of action of the enhancing factor could involve initiating and/or modulating steps of the leukocyte histamine release reaction . This factor, presumably a lymphokine or a monokine, may constitute a regulating link between cell-mediated immunity and histamine-mediated hypersensitivity reactions in allergic patients.

Eur J Immunol, 1978 Aug, 8(8), 607 - 11
Rat macrophage-mediated toxicity to cancer cells; effect of endotoxins and endotoxin inhibitors contained in culture media; Martin F et al.; The cytotoxic effect of normal or BCG-activated rat macrophages on a syngeneic line of cancer cells was compared in media containing rat or bovine sera . Normal macrophages were usually cytotoxic to cancer cells in fetal or newborn bovine serum; however, they enhanced cancer cell growth in normal rat serum . BCG- activated macrophages were toxic to cancer cells regardless of the serum used in the assay . Many cell culture media or sera obtained commercially were found to be contaminated by bacterial endotoxins . When endotoxin-free reagents were used in the toxicity assay, different results were observed: normal macrophages were not cytotoxic, but rather, they often enhanced cell growth even in fetal bovine serum; toxicity of activated macrophages was significantly reduced in normal rat serum . These results suggest that endotoxin and endotoxin inhibitors play a role in the modulation of macrophage-mediated cytotoxicity . These results emphasize the importance of monitoring endotoxin contamination in cell culture reagents used in assays involving macrophages.

J Biochem (Tokyo), 1978 Aug, 84(2), 453 - 60
Formation of biologically active protein from the subunits of islets-activating protein (IAP), a new protein isolated from Bordetella pertussis; Kanbayashi Y et al.; Based on the finding reported in the preceding paper (Kanbayashi, et al.: J . Biochem) that subunits of islets-activating protein (IAP), a new protein purified from the culture media of Bordetella pertussis, were inactive as such, but regained the original biological activities when recombined, the conditions required for recovery of the biological activities were studied . Essentially the same biological activities as the native IAP were recovered when the smallest subunit, F-3, was incubated with one of the other subunits, F-1 and F-2, at a pH of around 7, at temperatures below 30 degrees C and for longer than 12 h . During the incubation, association products were formed which were isolated by gel filtration as homogenous proteins that consisted of two subunits probably in a molar ratio of 1 : 1 . The native IAP (consisting of two IAP subunits including F-3) were equipotent in enhancing insulin secretory responses, in inhibiting epinephrine-induced hyperglycemia, in inducing leukocytosis and in increasing histamine sensitivity in experimental animals.

J Exp Zool, 1978 Aug, 205(2), 277 - 84
Wound healing in tadpole tailfin pieces in vitro; Derby A; Explants of tail fins from R . catesbeiana tadpoles undergo reepithelialization of their cut surfaces (healing) when cultured in vitro in Hanks' balanced salt solution at 22 degrees C . Healing is initiated early and closure of the wound is complete by 12 to 24 hours . Morphogenesis continues for several days as further reorganization and migration of epidermal cells from the regions adjacent to the wound margins take place . The addition of serum to the culture media improves the general appearance of these tissues and promotes healing . The rate of healing is affected by temperature . Tail fins maintained at 10 degrees C do not heal while fins maintained at 30 degrees and 37 degrees, although healing more rapidly than at 22 degrees, undergo progressive degeneration in culture . Epidermal cell movements were also studied in explants consisting of a combination of intact tail fin plus tail fin deprived of its epithelium . Rapid and extensive migration of epidermal cells from the intact tail fin across the collagen lamella of the stripped fin is observed.

J Natl Cancer Inst, 1978 Aug, 61(2), 431 - 6
Transmission electron microscopy surveillance of retroviruses in tissue culture cells prepared by the critical-point drying method; Iwasaki Y et al.; Tissue culture cells grown on grids were processed by the critical-point drying whole-cell method . With the use of a conventional transmission electron microscope operating at 100 kV, this technique permitted visualization of intracytoplasmic organelles of unsectioned whole cells . The morphology of type C virus in the process of budding and also in extracellular locations closely resembled that revealed in thin sections . Prelimininary results of virus surveillance of tissue culture cells prepared by this technique was corroborated by the levels of reverse transcriptase activity in culture media and by immunofluorescence staining of viral antigens on the cell surface.

J Biochem (Tokyo), 1978 Aug, 84(2), 443 - 51
Subunit structure of islets-activating protein (IAP), a new protein isolated from the culture media of Bordetella pertussis; Kanbayashi Y et al.; The subunit structure was studied of islets-activating protein (IAP), a new protein recently isolated from the culture media of Bordetella pertussis and possessing a unique action, i.e., potentiating insulin secretory responses of animals, IAP dissociated into three subunits, F-1, F-2, and F-3, when incubated in 8M urea . Three subunits isolated by chromatography on CM-Sepharose and DEAE-Sepharose columns showed different molecular weights (F-1: 44,000, F-2: 20,000, F-3: 11,000) and different isoelectric points, but similar amino acid compositions . The F-1 subunit consisted of two polypeptide chains linked by S-S bonding(s), while the F-2 and F-3 subunits were single-chain peptides . These subunits, none of which was biologically active alone, associated upon incubation for 2 h at 37 degrees C and regained biological activities after association only when the F-3 subunit was present in the association product . Thus, the F-3 subunit was essential, and the F-1 and F-2 subunits were permissive, for the development of IAP activity in animals.

Aktuelle Gerontol, 1978 Aug, 8(8), 403 - 9
Changes of glycosaminoglycan synthesis during in vitro ageing of human fibroblasts (WI-38); Schachtschabel DO et al.; Synthesis rates of glycosaminoglycans by WI-38 cultures (diploid, human fibroblasts exhibiting a limited number of population doublings in vitro) were determined by incorporation of 35S-sulfate of 14C-glucosamine into cellular and extracellular glycosaminoglycans at different passage levels before phase out . A progressive decline in the synthesis of cellular and extracellular glycosaminoglycans occured during the last (about 4) population doublings . 35S-sulfate incorporation into extracellular glycosaminoglycans appeared to be somewhat more reduced than 14-C-glucosamine incorporation during the last passages . Analysis of the distribution pattern of incorporated label into various glycosaminoglycan types (hyaluronic acid, chondroitin sulfate, dermatan sulfate and possibly heparan sulfate) revealed an age-related relatively stonger decline of 14C-glucosamine incorporation into cellular and extracellular hyaluronic acid and of 35S-sulfate into extracellular chondroitin sulfate in comparison with the other glycosaminoglycan types . Addition of exogenous glycosaminoglycans (chondroitin-4-sulfate, chondroitin-6-sulfate, dermatan sulfate, hyaluronic acid, heparan sulfate, heparin) at 100 microgram/ml to the culture media during the last 7 to 10 population doublings before phase out did not increase the total number of population-doublings . Heparin exhibited a significant growth inhibitory effect at 100 or 500 microgram/ml . The changes in glycosaminoglycan metabolism are interpreted as an expression of cellular ageing, and such an in vitro system offers a model for analyzing the factors involved in or causing the induction respectively prevention of this functional change.

Br J Obstet Gynaecol, 1978 Jul, 85(7), 551 - 6
Production of fibrinolytic enzymes by macrophages on intrauterine contraceptive devices; Foley ME et al.; Intrauterine contraceptive devices (IUCDs) were removed from 44 patients with a variety of clinical conditions, and incubated in culture media . Following incubation for up to 96 hours the total numbers of macrophages on each device were counted . The Lippes loop and Saf-T- Coil had higher counts than the Copper 7 . The counts on all devices were higher at mid-cycle and during menstruation and significantly higher in patients with menorrhagia and intermenstrual bleeding (P less than 0.0005) . Samples of culture media were taken on a number of occasions for up to 96 hours for fibrinolytic studies, and fibrinolytic activity increased with time in 10 of 16 cases where fibrinolytic activity was detected . There was a weak positive correlation between the number of cells on each of the 10 devices which produced a rise in fibrinolytic activity and the highest level of activity produced by each of the devices (r = +0.59; P less than 0.05) . Plasminogen activator activity was maximum early in the incubation period, while plasmin-like activity predominated in later samples . The possible role of macrophages in IUCDs in causing menorrhagia is discussed.

J Clin Endocrinol Metab, 1978 Jul, 47(1), 168 - 70
In vitro effect of dopamine and pimozide on human chorionic somatomammotropin (hCS) secretion; Macaron C et al.; In human placental explants cultured in vitro, dopamine inhibited human chorionic somatomammotropin (hCS) secretion into the culture media . In the control flasks, the level of hCS secretion was 130.5 +/- 7.8 micrograms/g tissue (n = 6) . When 1 mM dopamine was added, hCS levels decreased to 80.2 +/- 11.5 micrograms/g tissue (P less than 0.01) . Dopamine (5 and 10 mM) further lowered hCS levels . In contrast, 1 mM pimozide enhanced hCS secretion by 2-fold as compared to control levels (248.2 +/- 44.8 vs . 130.5 +/- 7.8, P less than 0.02) . The simultaneous addition of dopamine did not alter the stimulatory effect of pimozide on hCS secretion . In separate experiments, arginine (1 and 5 mM) and somatostatin (1 microgram/ml culture media) did not alter hCS secretion from placental explants . These results suggest that hCS secretion is modulated by dopaminergic receptors.

Diabetologia, 1978 Jun, 14(6), 397 - 404
Isolated mouse pancreatic islets in culture: effects of serum and different culture media on the insulin production of the islets; Andersson A; Various conditions for tissue culture of collangenase-isolated mouse pancreatic islets were studied in an attempt to optimize the maintenance of glucose stimulated insulin biosynthesis and release in the cultured specimens . Islets which had been cultured at a physiological glucose concentration (5.5 mmol/1) in the absence of serum had an impaired glucose-stimulated insulin biosynthesis and release as well as a reduced insulin content . Thus, insulin biosynthesis was three times higher after culture in a serum supplemented medium . Further, the insulin secretion of islets cultured in the presence of serum was markedly enhanced in acute incubations with high concentrations of glucose . This response was most pronounced in islets which had been cultured free-floating . A comparison between different culture media showed that islets cultured in RPMI 1640 had the highest insulin production . The present data suggest that the most favourable conditions for long-term storage of isolated islets in culture may be obtained when the islets are maintained as free-floating explants in a culture medium consisting of RPMI 1640 supplemented with serum.

J Exp Med, 1978 Jun 1, 147(6), 1551 - 67
Requirement for hexose, unrelated to energy provision, in T-cell-mediated cytolysis at the lethal hit stage; MacLennan IC et al.; The requirement for D-glucose in T-cell-mediated cytolysis was studied using mouse spleen cells sensitized against alloantigens in vitro . Glucose was required for cytolysis: (a) cytolysis proceeded in a simple buffered salt solution containing Ca++ and Mg++ (low phosphate-buffered saline, LPBS) in the presence but not in the absence of added glucose; (b) 2-deoxy-D-glucose blocked cytolysis . The block by this agent was overcome by excess glucose added as late as 40 min after the inhibitor . This block was not due to inhibition of NADP reduction, since 2-deoxy-D-glucose failed to interfere with the rate of CO2 production by the pentose cycle which we found to be of significant activity in sensitized spleen cells; (c) dialyzed fetal bovine serum (DFBS) in LPBS supported cytolysis in the absence of added glucose . However, 2-deoxy-D-glucose was also inhibitory under these conditions, suggesting that carbohydrate was required here as well . Further results supported the conclusion that DFBS was not acting as a direct source of the required carbohydrate . The relationship between cytolysis, glucose requirement, and provision of energy was studied . As little as 0.1 mM D-glucose in LPBS supported cytolysis . At this glucose concentration, there was no measurable accumulation of lactate in sensitized spleen cells, but Krebs cycle activity was detectable . In 3 mM glucose or above, the range covered by standard tissue culture media, anaerobic glycolysis became a major source of energy in sensitized spleen cells . Consequently, it appears that in standard tissue culture medium, effector cells can generate sufficient energy for cytolysis either by aerobic or anaerobic metabolism . However, the addition of an energy source alone in the absence of glucose was insufficient to support cytolysis in LPBS . Pyruvate in LPBS did not support cytolysis but was shown to be a good substrate for aerobic metabolism in sensitized spleen cells . Glycogenic amino acids and glycerol also failed to support cytolysis . The stage of cytolysis at which glucose is required was investigated . Glucose was necessary for the calcium-dependent lethal hit phase, but not for the cytochalasin A-blockable recognition stage, nor for 51Cr release from injured target cells . Models for the lethal hit process are discussed, which are compatible with the observed requirement for certain hexoses unrelated to their capacity to serve as sources of energy.

Proc Natl Acad Sci U S A, 1978 Jun, 75(6), 2621 - 5
Synthesis of cold-insoluble globulin by cultured calf endothelial cells; Macarak EJ et al.; Radiolabeled amino acids were incorporated into nondialyzable protein by cultures of endothelial cells derived from calf aorta . Antibody prepared against purified bovine plasma cold-insoluble globulin (CIG) formed a strong precipitin line upon immunodiffusion against 3H-labeled proteins from endothelial cell culture media . This precipitin line formed a line of identity with the precipitin line formed by anti-CIG and purified CIG . Double-antibody immunoprecipitation of 3H-labeled protein showed that 36.3% of the radioactivity was specifically precipitated by the anti-CIG antibody . Gel filtration and polyacrylamide gel electrophoresis of the immunoprecipitate indicated that it contained a single protein species whose molecular weight was consistent with that of CIG isolated from plasma . When cultured cells were examined by indirect immunofluorescence microscopy, the endothelial cells stained specifically with anti-CIG antibody . These data indicate that cultured endothelial cells derived from calf aorta synthesize and secrete a protein antigenically similar to CIG.

J Urol, 1978 Jun, 119(6), 777 - 9
The effect of human serum on 3H-thymidine incorporation in human prostate tumors in tissue culture; Boileau M et al.; Human prostatic tumors, 4 adenomas and 4 adenocarcinomas, were established in tissue culture . We used the incorporation of 3H-thymidine into deoxyribonucleic acid as a measure of cell activity to compare the effects of supplementing the culture media with human or fetal calf serum . 3H-thymidine incorporation was greater with human serum, with up to 120 per cent increase observed . The addition of dihydrotestosterone (10(-8)M.) did not enhance or inhibit 3H-thymidine incorporation.

Biochem J, 1978 May 15, 172(2), 275 - 84
Degradation of cartilage proteoglycans by a neutral proteinase secreted by rabbit bone-marrow macrophages in culture; Hauser P et al.; When cultivated together with pieces of cartilage biosynthetically labelled with 35S in their proteoglycans, rabbit macrophages, differentiated in vitro from bone-marrow cells, cause the release of soluble 35S-labelled material into the culture medium . This process is inhibited by killing the macrophages or by cycloheximide treatment, and is due to the secretion by the cells of a metal-dependent neutral proteinase capable of degrading cartilage proteoglycan subunits into fragments of high molecular weight . Enzyme activity is optimum at about pH7, and is inhibited by EDTA, o-phenanthroline, cysteine or serum, but not by di-isopropyl phosphorofluoridate nor by 4-hydroxymercuribenzoate . The effect of EDTA is partially reversed by Co2+ or Zn2+ ions . The enzyme is eluted from Sephadex G-150 columns as a single peak of material (apparent mol.wt . 17000) that contains also most of the proteolytic activity exerted by culture media on Azocoll (denatured collagen) or on casein . The possible role of this metalloproteinase in chronic inflammatory processes is discussed, particularly in connection with joint erosions in rheumatoid arthritis.

Biochem J, 1978 May 15, 172(2), 261 - 74
The simultaneous release by bone explants in culture and the parallel activation of procollagenase and of a latent neutral proteinase that degrades cartilage proteoglycans and denatured collagen; Vaes G et al.; 1 . A latent neutral proteinase was found in culture media of mouse bone explants . Its accumulation during the cultures is closely parallel to that of procollagenase; both require the presence of heparin in the media . 2 . Latent neutral proteinase was activated by several treatments of the media known to activate procollagenase, such as limited proteolysis by trypsin, chymotrypsin, plasmin or kallikrein, dialysis against 3 M-NaSCN at 4 degrees C and prolonged preincubation at 25 degrees C . Its activation often followed that of the procollagenase present in the same media . 3 . Activation of neutral proteinase (as does that of procollagenase) by trypsin or plasmin involved two successive steps: the activation of a latent endogenous activator present in the media followed by the activation of neutral proteinase itself by that activator . 4 . The proteinase degrades cartilage proteoglycans, denatured collagen (Azocoll) and casein at neutral pH; it is inhibited by EDTA, cysteine or serum . Collagenase is not inhibited by casein or Azocoll and is less resistant to heat or to trypsin than is the proteinase . Partial separation of the two enzymes was achieved by gel filtration of the media but not by fractional (NH4)2SO4 precipitation, by ion exchange or by affinity chromatography on Sepharose-collagen . These fractionations did not activate latent enzymes . 5 . Trypsin activation decreases the molecular weight of both latent enzymes (60 000-70 000) by 20 000-30 000, as determined by gel filtration of media after removal of heparin . 6 . The latency of both enzymes could be due either to a zymogen or to an enzyme-inhibitor complex . A thermostable inhibitor of both enzymes was found in some media . However, combinations of either enzyme with that inhibitor were not reactivated by trypsin, indicating that this inhibitor is unlikely to be the cause of the latency.

J Med Microbiol, 1978 May, 11(2), 187 - 91
Comparison of several culture media used for studies on mycobacteriophages; Stager CE et al.; The value of RVA, N-1, 7H10, 7H11 and Sauton's media for studies on mycobacteriophage infeciton and lysis of mycobacteria was assessed . Experiments were made with mycobacteriophages BGI, BKI, CRI-3, G37, and LG, all of which lyse Mycobacterium smegmatis strain 607B, and with mycobacteriophage DS6A which lyses Mycobacterium tuberculosis strain H37Rv . The methods involved "direct lysis", the measurement of "routine test dilutions" and counts of plaque-forming units . It was found that N-1, 7H10 and 7H11 media gave better overall results than RVA medium for M . smegmatis strain 607B and its phages, and that RVA medium was generally the most useful for M . tuberculositems employed.

Arch Microbiol, 1978 Apr 27, 117(1), 49 - 52
The role of vanadium in green plants . IV . Influence on the formation of delta-aminolevulinic acid in Chlorella; Meisch HU et al.; In a series of experiments, it is demonstrated that the trace element vanadium (4.10(-7) g-at/1 as NH4VO3) has a considerable positive influence on the synthesis of delta-aminolevulinic acid(delta-ALA) in the autotrophically growing green alga Chlorella pyrenoidosa, the effect being visible by an enhanced output of the amino acid into the culture medium in presence of levulinic acid (LA) . The level of intracellularly accumulated delta-ALA, however, is not changed in presence of the metal . The V-effect on exogenous found delta-ALA is suppressed, when LA is added to the nutrient medium at low pH (pH5), although V-uptake into the algal cells is not disturbed by LA . As demonstrated in culture media with various nitrogen sources (urea, partially hydrolized urea, ammonium salts), the development of the pH during the cultivation time is important for the presentation of the V-effect on delta-ALA . It is suggested that vanadium acts as a catalyst in the conversion of 4,5-dioxovaleric acid to delta-ALA by transamination.

Am J Vet Res, 1978 Apr, 39(4), 687 - 90
Comparison of whole blood and purified canine lymphocytes in a lymphocyte-stimulation microassay; Shifrine M et al.; The optimum mitogen concentration and time required for using whole blood from dogs in a microassay were determined, and this test then was compared with a standard lymphocyte-stimulation microtest, using gradient-isolated lymphocytes, 2 different mitogens (phytohemagglutinin and concanavalin A), and 2 different culture media . Statistical analysis of the data from 10 dogs showed that whole blood was significantly more reactive than were gradient-isolated lymphocytes (P less than 0.05) . Waymouth's medium was significantly better than RPMI 1640 (P less than 0.001), and concanavalin A was significantly more mitogenic than phytohemagglutinin (P less than 0.001) . The interaction between lymphocyte source and mitogens was the only one of the various interactions that was significant at P less than 0.05.

Cytobiologie, 1978 Apr, 16(3), 381 - 92
Dextran T 500--induction of spreading in Ehrlich ascites tumour cells on glass surface; Cieslak J et al.; The experiments were carried out in order to find factors which could induce attachment and spreading of Ehrlich ascites tumour (EAT) cells on solid substrata . In normal culture media, serum-free as well as serum-containing, these cells did not spread and very weakly attached onto glass . It was found that after coating the cell surfaces with dextran T 500 the EAT cells strongly attached and spread extensively on glass . This spreading could be inhibited or reversed by washing out the dextran or adding calf serum . Dextran T 500 caused rapid spreading also in chick embryo fibroblasts and mouse lymphocytes . Some aspects of these results in connection with contemporary views concerning the processes of cell attachment and spreading are discussed.

J Parasitol, 1978 Apr, 64(2), 283 - 9
Phenol oxidase activity: inductionin female schistosomes by in vitro incubation; Seed JL et al.; A biochemical methods has been developed for detecting phenol oxidase in female Schistosoma mansoni . Enzyme activity is observed only after incubation of the female schistosomes for an extended period of time in tissue culture media . Male S . mansoni do not contain detectable levels of phenol oxidase activity . The properties of this enzyme are similar to those identified for a phenol oxidase from Fasciola hepatica . L-DOPA, dopamine, and tyrosine were found to be good substrates for this enzyme . Vmax = 14.1, 8.1, and 6.1 mumoles O2/min/mg protein for each substrate, respectively . This enzyme appears to be associated with egg production and thus may be a useful marker for biochemical and immunological studies.

J Cell Physiol, 1978 Apr, 95(1), 49 - 55
Thymidylate synthetase and dihydrofolic acid reductase in the stimulated human lymphocyte; Haurani FI et al.; The phytohemagglutinin (PHA)-stimulated human lymphocytes demonstrated trace or no activity (dihydro) folate acid reductase using three methods including a radioassay, but demonstrated ample activity of thymidylate synthetase . This was true regardless of the day of harvest, (first through seventh) of the stimulated lymphocyte . The lymphocyte extracts revealed no inhibitor to the reductase enzyme when these extracts were added before the liver extracts to the assay system . When methotrexate (MTX) was added to the culture media of the lymphocytes, there was, as expected, an increase in the synthetase activity, but the expected rise in the reductase activity did not occur, it remained nil . On the other hand, MTX did influence the incorporation of nucleosides by the stimulated lymphocytes in a fashion similar to its action on the incorporation of the same nucleosides by other cells.

J Natl Cancer Inst, 1978 Apr, 60(4), 773 - 7
Purification and immunologic evaluation of human melnoma-associated antigens; McCabe RP et al.; Melanoma-associated antigens (MAA) were isolated and their functional immunologic properties were evaluated . Spent fetal calf serum-free culture media and 3-m KCI extracts of cultured human melanoma cells grown in this medium were used as antigen sources . Ultracentrifugal flotation on KBr was used to separate MAA and HLA antigens present in the extracts or spent culture media; thus interference by histocompatibility antigens was prevented in subsequent tests of tumor antigenic activity . MAA purified in this manner retained their immunologic functions as evidenced by their ability to produce delayed cutaneous hypersensitivity reactions in patients with melanoma, specifically combine with antimelanoma xenoantibody, and elicit production of functionally specific xenoantibody . Possible structural differences between HLA antigens and MAA were considered in evaluation of the data.

Z Parasitenkd, 1978 Mar 16, 55(1), 63 - 72
{Experiments in vitro with Litomosoides carinii (Nematoda: Filarioidea) . I . Maintenance of adult females and microfilariae as well as release of microfilariae in different culture media (author's transl)}; Wenk P et al.; Embryos of L . carinii continue intrauterine development to microfilariae and are totally released into the medium within 5--6 days when the latter (Tc 199) is changed daily and air is used as the gas phase . Oogenesis or further fertilization of eggs, however, does not occur in vitro in any of the media examined by us . One female releases 140 X 10(3) microfilaria/day on an average in vitro within 5--6 days . Mean initial numbers of 300 X 10(3) Mf/female/day are observed . Addition of equine serum inhibits microfilarial release in vitro; normal cotton rat serum prolongs survival of females while total numbers of released microfilariae or retained embryonic stages are not increased . The serum of post-patent animals does not influence the numbers of released microfilariae or their viability or survival of females . Microfilariae released in vitro in Tc 199 + 33% normal cotton rat serum survive for more than 8 days, when air is used as the gas phase and the medium is changed daily . Microfilariae isolated from the blood of patent animals survive for at most 6 days, at a 48-hourly change of medium survival does not even exceed 4 days.

Growth, 1978 Mar, 42(1), 59 - 69
Canine kidney cells: I . Neutral lipids, phospholipids and fatty acids of SV40 transformed canine kidney cells in suspension and monolayer culture; Hougland AE et al.; Differences in lipid composition and the effects of monolayer and suspension culture on SV40 virus transformed canine kidney cells and their plasma membranes were studied . Plasma membranes from monolayer and suspension cultures of SV40 transformed canine kidney cells were isolated by the Warren fluorescein-mercuric acetate technique . Free cholesterol was the major neutral lipid extracted from the whole cells and plasma membranes of all these cultures . Triglyceride increased about 6- to 50-fold in suspension grown whole cells and their plasma membranes compared to those from monolayer cultures . The mu moles of total neutral lipids and phospholipids are elevated in plasma membranes compared to whole cells from monolayer cultures but are similar in suspension cultures . The distribution of fatty acids in the major lipid classes of plasma membranes and whole cells generally follow the composition of calf serum used in the culture media . INDEX WORDS: SV40, plasma membranes, transformed cells, canine kidney cells, neutral lipids, phospholipids, monolayer culture, suspension culture.

J Clin Pathol, 1978 Feb, 31(2), 162 - 4
Synergy between sulphonamide and trimethoprim in the presence of pus; Edmunds PN; Synergy between sulphadiazine and trimethoprim against Escherichia coli has been demonstrated in culture media not containing lysed horse blood despite the presence of pus or a pus extract which inhibited the action of each drug separately . Synergy may therefore be important where pus is present in vivo.

Endocrinology, 1978 Feb, 102(2), 509 - 18
Enriched populations of rat pituitary thyrotrophs in monolayer culture; Leuschen MP et al.; Rat anterior pituitary cells were dissociated and subjected to 4 h of unit gravity sedimentation . Eighty-five 8-ml fractions were collected, and nine pooled fractions were placed in monolayer culture for 1 week . The attached cells were immunocytochemically stained for TSH using an antiserum previously shown to be specific for the beta subunit of TSH and all culture media saved for subsequent assay of TSH . Thyrotrophs were localized both immunocytochemically and by radioimmunoassay to 1-2 fractions from the top of the sedimentation chamber . Typically, it was found that 60-80% of the cells in these fractions were immunocytochemically identified as thyrotrophs . Electron microscopic observations indicated that such cells displayed the classical morphological features associated with thyrotrophs . The enriched thyrotroph cultures responded to 1.0 mM dibutyryl cyclic AMP with an increased release of TSH indicating that the intracellular secretory machinery was not altered by the procedures employed . In all cases, the basal TSH secretion decreased with time in culture . In addition, the fractions containing the most TSH in the culture media were usually one fraction lighter (lower sedimentation rate) than the fractions which were found immunocytochemically to contain the highest percentage of thyrotrophs . The results suggest the possibility of two functionally distinct thyrotroph cell types as has been suggested for the pituitary somatotrophs.

Exp Cell Biol, 1978, 46(6), 325 - 37
Replication of mouse mammary tumor cells in monolayer cultures stimulated with embryo extract; Baumann KR et al.; A particulate fraction derived from homogenized chicken embryos was added to primary monolayer cultures of mouse mammary tumor cells . All culture media contained fetal calf serum (10%) . After 4 days, the extract had stimulated a twofold increase (relative to controls lacking the extract) in (1) total DNA per culture; (2) the rate of incorporation of labeled DNA precursors; (3) epithelial cell number, and (4) total protein . This integrated growth activity is discussed in terms of the nature and exogenous regulation of neoplastic growth.

Ann Biol Clin (Paris), 1978, 36(1), 23 - 5
{Rabies virus study by the plaque forming system . A simplified technique}; Salaun JJ; The author describes here a simplified and economical plaque forming system for rabies virus study . It is achieved in three steps (cells preparation and infection, apply of overlay, staining) . It presents more advantages than previous techniques: time as well as culture media and handling spearing, easy and durable reading . Cells prepared with di-ethyl-amino-ethyl-dextran are added to equal quantities of diluted virus (for titration) or to virus-serum mixing (for seroneutralization) and overlaid after 4 hours with a carboxymethylcellulose medium . Reading is carried out after 6 days incubation and staining with amido black . Plaques obtained are to 2-3 mm in diameter, regularly reproductible, clear and easy to read . This technique enables titration and seroneutralization and even a rabies virus cloning.

J Immunol, 1978 Jan, 120(1), 96 - 101
Antibody response and tumor growth in syngeneic mice immunized to partially purified B16 melanoma-associated antigens; Bystryn JC; Soluble murine melanoma-associated antigens (MAA), partially purified from the media of B16 melanoma cells in culture by ammonium sulfate precipitation and Sephadex G-200 chromatography, were tested for their effect on tumor growth . MAA were immunogenic in syngenic mice as evidenced by their ability to induce anti-melanoma antibodies and partial protective tumor immunity . The level of immunity was variable . It ranged from retardation of tumor development to almost complete suppression of tumor growth . The results were influenced by the nature of the control group, since immunization to either normal tissue antigens or complete Freund's adjuvant enhanced tumor growth . Overall, 46 of 91 MAA immunized mice, but none of 114 control mice, survived over 6 weeks (p less than 0.001) . The protective immunity was specific since MAA immunized mice were not resistant to challenge with an unrelated syngeneic tumor (BW 10232 ADENOCARCINOMA) . Partially purified normal tissue antigens were also immunogenic in syngeneic mice . They induced low levels of antibodies to, and enhanced the growth of, melanoma . These findings indicate that the culture media of melanoma cells contains tumor antigens that retain their biologic activity after partial purification, and that can induce specific, although only partial, protective immunity to melanoma.

Immunol Commun, 1978, 7(5), 557 - 66
Effect of delta9-tetrahydrocannabinol on in vitro sensitization of mouse splenic lymphocytes; Lefkowitz SS et al.; The effects of delta9-tetrahydrocannabinol (delta9-THC) on the immune response of murine cells sensitized in vitro was determined using a plaque-forming cell (PFC) assay . Splenic lymphocytes from mice injected with delta9-THC showed a depressed immunologic response when compared with cells from control animals which were identically sensitized in vitro with sheep erythrocytes (SRBC) . The direct addition of delta9-THC to the culture media altered the immunological response as demonstrated by a reduction in the number of PFC.

Proc Natl Acad Sci U S A, 1978 Jan, 75(1), 459 - 63
Anti-epidermal-cell-surface pemphigus antibody detaches viable epidermal cells from culture plates by activation of proteinase; Farb RM et al.; Immunoglobulin from pemphigus patients binds to the surface of mouse epidermal cells in culture . Cells incubated with the pemphigus antibody are easily detached from culture plates whereas cells incubated with serum from normal patients remain on the plate . Pemphigus antibody-mediated cell detachment is blocked by the addition of the proteinase inhibitors soybean trypsin inhibitor and alpha2-macroglobulin to the culture media . Detachable cells are viable, and activation of the complement cascade is not necessary for cell detachment . The anti-cell-surface antibody of pemphigus appears to disrupt adhesion between viable epidermal cells by activation of proteinase.

Ciba Found Symp, 1978, (64), 33 - 52
Changes in the surface of the mouse blastocyst at implantation; Sherman MI et al.; Implantation is a critical event, and perhaps the earliest one, in the maternal recognition of pregnancy . Information transfer from conceptus to mother might occur during, and subsequent to, implantation at the level of cell surface interaction . Therefore, attempts have been made both to identify the phases of implantation during which changes in the blastocyst surface occur and to characterized such changes . In vitro, blastocysts have been found to go through a series of discrete steps which are analogous to implantation in utero, and these steps can be retarded or prevented by the use of either suboptimal culture media or an inappropriate substratum . Morphological surface changes are not apparent when the blastocyst becomes adherent to the substratum; however, marked differences in blastocyst surface structure are revealed by scanning electron microscopy at the onset of trophoblast outgrowth . Studies at the molecular level implicate collagen as having a role in blastocyst adhesiveness, but other cell surface components are also likely to be involved.

Rev Ig Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Pneumoftiziol, 1978 Jan-Mar, 27(1), 53 - 8
{The effectiveness of some culture media with pyruvic acid bases for the isolation of mycobacteria from sputum}; Timosca S et al.; The authors have studied the value of culture media based on pyruvic acid (the Dixon medium and an original medium called P.T.) in the isolation of mycobacteria from sputum, as compared with the Lowenstein-Jensen medium . The studies were carried out in two groups of patients: 1043 samples were collected from ambulatory patients and 1740 samples were collected from hospitalized patients . The superiority of the Dixon and of the P.T . media was confirmed by the increased percentage of positive cultures (11% and 20% respectively) as compared with the Lowenstein-Jensen medium . With these media it was possible to isolate mycobacteria 7--14 days earlier than with the other media . On the basis of the results obtained the authors recommend the use, in parallel, especially in those cases when the samples contain only a small number of bacteria, of the Lowenstein-Jensen medium and one of the media based on pyruvic acid.

Acta Microbiol Pol, 1978, 27(3), 237 - 42
Effect of various nitrogen salts in beet pulp medium on polygalacturonase activity of Penicillium sp . 7/4B/EI 1 mutant; Szajer I; The highest PG activity was obtained in beet pulp with (NH4)2HPO4 in amount equivalent to 0.14--0.28% N after 7 days of growth; At optimal concentration, (NH4)2HPO4 did not influence the biomass production, but increased the pH of the culture media to above 6.0 . The PG was an extracellular enzyme, acting mainly on highly esterified pectin . The attack was of a random manner, characteristic for the endo-enzymes . The enzyme was active at temperature 20 degrees C--40 degrees C and pH 5.0--6.0.

Int J Cancer, 1977 Dec 15, 20(6), 817 - 23
Increase in immunogenicity of a pulmonary squamous-cell carcinoma, propagated in vitro; Jamasbi RJ et al.; The chemically induced, non-immunogenic lung squamous-cell carcinoma (MSC-10) propagated in vitro gradually loses tumorigenicity in immunocompetent hosts with increasing in vitro passage . This was found to be related to an increase in antigenicity, since immunosuppressed hosts (thymectomy plus 600 rads whole body X-irradiation) supported the growth of tumor cells, whereas immunocompetent recipients did not . The antigens involved in rejection are not heterologous serum proteins present in culture media since the cell line grown in isologous serum is also rejected . Immunization with the in vitro tumor line partially protected against the parental in vivo line, therefore the antigens involved must be present on both tumor lines . Inoculation of the cultured cell line into normal or immunosuppressed hosts produced tumors with the same histological characteristics as those of the in vivo tumor line . We concluded that by in vitro culture the weakly antigenic carcinoma becomes more immunogenic and thereby capable of inducing transplantation resistance . The cultured tumor cells retain their antigenic specificity and histologic characteristics while increasing their antigenic potency.

J Clin Endocrinol Metab, 1977 Dec, 45(6), 1271 - 80
Immunoreactive luteinizing hormone, follicle stimulating hormone and their subunits in tissue culture media from normal and adenomatous, human pituitary fragments; Beitins IZ et al.; Fragments from 4 human pituitaries removed at surgery from one gonadectomized man and three women (one with Nelson's syndrome, one with Forbes Albright syndrome and one with chromophobe adenoma) were grown in tissue culture . The tissue culture medium was changed at weekly intervals, pooled and applied to a Sephadex G-100 column for gel filtration . In each eluate luteinizing hormone (LH), follicle stimulating hormone (FSH), alpha subunit and the beta subunits of LH and FSH were measured by specific radioimmunoassays . For comparison the pituitary standard LER 907 was similarly studied . LH and FSH were measurable in all studies . LH eluted over a broad area whereas FSH eluted as a much sharper peak . In all culture media and in LER 907 large quantities of alpha subunit were detected . The beta subunits of LH and FSH were not present in the LER 907 standard . LHbeta subunit was present in the culture medium of the pituitary fragments from the castrated man and from the women with Nelson's syndrome and Forbes Albright syndrome but not in that of the woman with chromophobe adenoma . FSHbeta subunit was detectable only in the latter case.

Cancer, 1977 Nov, 40(5 Suppl), 2699 - 2705
Restoration of in vitro growth control to malignant cells; Lipkin G et al.; A glycoprotein (molecular weight, ca . 160,000) from culture medium of contact-inhibited hamster melanocytes restores contact inhibition of growth to malignant melanocytes of man, mouse, and hamsters, and also effectively inhibits growth in vitro of a broad spectrum of malignant and normal cell types of ectodermal, mesodermal and endodermal origins, including human colon carcinomas . The melanocyte contact inhibitory factor (MCIF) produces G1 growth arrest in malignant melanocytes; inhibition of all cell types is reversible, does related, and nontoxic at concentrations below 200 microgram/ml, but selectively lethal to malignant cells at higher concentrations . An electrophoretically identical protein is present in culture media of contact-inhibited melanocytes, fibroblasts, and epidermal cells, but absent from those of colon carcinomas, HeLa cells and malignant melanomas . Nevertheless, an MCIF-like band is present in whole cell homogenates of human colon carcinomas and hamster melanomas . MCIF may permit normal surface interactions required for feedback inhibition of growth.

Br J Cancer, 1977 Nov, 36(5), 601 - 7
Secretion of prostaglandins as bone-resorbing agents by renal cortical carcinoma in culture; Atkins D et al.; Fragments of human renal carcinoma tissue have been co-cultured with mouse calvaria . In 9/13 cases significant bone resorption occurred whilst in no case did control kidney cause significant resorption . When bone resorption did occur, it could be reduced by inclusion of indomethacin in the culture medium . In some cases when theophylline was included in culture medium to prevent cyclic AMP breakdown, there was enhancement of tumour-induced bone resorption . Control studies without tumour showed that none of the experimental treatments had a direct effect on bone . Radioimmunoassay of prostaglandin E (PGE) levels in pooled culture media showed that tumour fragments produced appreciable amounts of PGE, and that this production was lowered by indomethacin and increased by theophylline . It is concluded that the bone resorption induced by these tumours is due to a prostaglandin, and that prostaglandin production may be controlled by changes in cyclic AMP metabolism.

Proc Natl Acad Sci U S A, 1977 Nov, 74(11), 4955 - 8
Prostaglandin regulation of macrophage collagenase production; Wahl LM et al.; The production of collagenase (EC 3.4.24.3) by endotoxin-stimulated macrophages was significantly inhibited by indomethacin, indicating that prostaglandins (PGs) mediate this effect . Inhibitions of collagenase production by indomethacin was overcome by addition of exogenous PGE2 at 10 nM whereas the addition of 0.1 and 1.0 micrometer PGE2 increased the enzyme production to 3 times that achieved by endotoxin . Although the addition of prostaglandin alone to macrophage cultures did not stimulate collagenase production, the simultaneous addition of PGE1 or PGE2 and endotoxin enhanced collagenase activity 2- to 10-fold . This increase was detectable at PGE concentrations of 10 nM and was optimal at 0.1-1.0 micrometer . PGF2alpha had little effect on either the enhancement of collagenase production by endotoxin-stimulated macrophages or its restoration in cultures inhibited by indomethacin . Radioimmunoassay of prostaglandins in the culture media revealed that macrophages exposed to endotoxin secreted 40-fold more PGE2 than did unstimulated cells . The increase in PGE2 was detected 4 hr after exposure to endotoxin and was maximal at 14 hr . The peak PGE2 concentrations in the culture media were similar to those of exogenous PGE2 (about 10 nM) needed to restore collagenase production in indomethacin-treated cultures . These findings demonstrate the involvement of PGE in the endotoxin-activation of macrophages with resultant production of collagenase.

Virchows Arch B Cell Pathol, 1977 Oct 27, 25(2), 171 - 7
Effects of erythrocyte lysate and erythrocyte-conditioned medium on erythroid cells in vitro; Bateman A et al.; A component of erythrocyte-conditioned medium has been shown to inhibit the incorporation of tritiated thymidine into erythroblasts from fetal mouse liver, proliferating in vitro . This component, however, has no detectable effect on the growth of colonies of erythroid cells stimulated to grow in viscous culture media by the hormone erythropoietin . Erythrocyte lysate and preparations of haemoglobin derived from the lysate increase the number and size of the colonies growing in vitro . Results are discussed in terms of possible control mechanisms in erythropoiesis.

No Shinkei Geka, 1977 Oct, 5(11), 1135 - 41
{Growth hormone, prolactin, LH and FSH secretion in tissue culture of pituitary adenomas (author's transl)}; Kubo O et al.; We have performed tissue culture of pituitary adenomas (5 acromegalies, 7 non-acromegalies) and measured GH, PRL, LH and FSH in the media of tissue culture by radioimmunoassay (RIA) for 30-40 days . In addition, GH, PRL, LH and FSH were measured in the plasma and tumor tissue by RIA . GH concentration in the culture media was markedly high in all cases with acromegaly and high also in 2 out of 7 non-acromegaly cases (case 6 and 7) . GH concentration in the media rapidly decreased almost as a straight line on a semilogarithmic scale until the 30-40 in day, when GH level became less than 10 ng/ml . PRL concentration in the culture media was high in 5 patients (3 acromegalies, 2 non-acromegalies) . PRL concentration regressed more slowly than GH, but still remained higher than 10 ng/ml in all of the patients on the 25-40th day when the studies were discontinued . LH and FSH concentrations in the culture media was high in two patients . One patient was a pituitary adenoma secreting LH and FSH, and another case was a false high secreting one, possibly contaminated with gonadotropin in normal pituitary gland at surgical operation . LH and FSH concentrations rapidly decreased as that of GH . A rare case of primary LH and FSH secreting pituitary adenoma was demonstrated.

J Clin Microbiol, 1977 Oct, 6(4), 362 - 6
Comparative evaluation of different types of blood culture media for isolation of aerobes; Gross PA et al.; The possible advantage of hypertonic sucrose medium over isotonic medium for isolating aerobic organisms from blood was studied . Approximately 50 ml of medium and 5 ml of blood inoculum were present in each culture bottle . In the first phase of the study, supplemented peptone broth (SPB) was compared with brucella broth containing 10% sucrose (BB-10S) . There were 194 significant clinical isolates in at least one of the two bottles in each set during a 7-month period; 160 (82%) of the isolates grew in SPB, whereas 191 (98%) grew in BB-10S (P less than 0.01) . Of the 158 isolates that grew in both media, 13 (8%) appeared earlier in BB-10S, whereas none did so in SPB . In the second phase of the study, SPB with 10% sucrose (SPB-10S) was compared with BB-10S . There were 187 isolates during a 9-month period; 173 (93%) grew in SPB-10S compared with 179 (96%) for BB-10S . In this comparison there was no apparent difference in the time interval required for recovery of organisms . The two hypertonic sucrose media (SPB-10S and BB-10S) were comparable for isolating organisms under aerobic conditions and superior to the nonhypertonic medium (SPB).

Biomedicine, 1977 Oct, 26(5), 344 - 9
Effects of serum-free culture media on human liver and fibroblastic cells; Lemonnier F et al.; The morphological aspects and amino acid variations of human fibroblast and liver cell monolayer cultures were studied in serum free media . Under these conditions, the behaviour of the two cell types differed greatly . The morphological changes for the liver cells, as compared with the fibroblasts, appear more quickly and some of these changes are particular . Furthermore, extracellular amino acid variations are of lesser importance for the liver cells, during the 10 days following the suppression of serum, except for serine.

Clin Chim Acta, 1977 Oct 1, 80(1), 105 - 11
Ectopic production of a salivary type amylase by adenocarcinoma cells: demonstration by a culture technique; Leda M et al.; Characterization of an elevated amylase activity in ascitic fluid obtained from a patient with carcinomatous peritonitis is described; ninety-one percent of the increased amylase activity in the fluid was of salivary type and the remainder of pancreatic type, when studied by ion-exchange chromatography . Culture of ascitic cells successfully demonstrated morphologically characterized tumor cells surviving for at least 31 days . During that period, significant amylase activities were detected in the culture media, and the isozyme pattern was a single band whose electrophoretic mobility corresponded to salivary amylase . The data obtained indicate that the ascites amylase of salivary type was produced ectopically by the tumor cells.

Steroids, 1977 Oct, 30(4), 569 - 80
Short term culture of human midterm and term placenta: parameters of hormonogenesis; Hall CS et al.; Monolayer cultures of human midterm and term placentae have been established following trypsin dispersion of placental minces . Maintenance of endocrine function was monitored by the concentrations of specific hormones in the culture media . At either gestational age the cultures 1) secret estradiol-17beta(1) and estrone (in a ratio of about 1:20) and aromatize 3H- or 14C-dehydroepiandrosterone sulfate and 14C-androstenedione, estrogen production being markedly enhanced by addition of dehydroepiandrosterone (10(-6)7) to the culture medium; 2) metabolize 3H-pregnenolone to progesterone and 14C-cortisol to cortisone; and 3) produce increasing amounts of chorionic gonadotropin and decreasing amounts of placental lactogen during the first week in culture . It is proposed that the model is highly suited to the study of factors affecting hormonogenesis by the human placenta whether they be of maternal or of fetal origin.

J Biol Chem, 1977 Sep 25, 252(18), 6310 - 5
Pseudouridine-deficient transfer RNAs from Escherichia coli B and their use as substrates for pseudouridine synthetase; Kwong LK et al.; Transfer RNAs isolated from Escherichia coli B grown in the presence of 2-thiouracil are deficient in pseudouridine . Much of this deficiency is from the T psi C region, which has only about 50% of its normal pseudouridine content . The other modified nucleoside from this region, ribothymidine, is reduced by only about 10% . Studies showed that 2-thiouracil is incoproated into the RNA of E . coli during growth in the presence of the analog . This incorporation appears to result from the replacement of uracil, occur in a random manner, and involve all RNA species . The extent of incorporation varies from 1 to 3 mol %, depending upon the preparation and RNA species examined . Electrophoresis on polyacrylamide gels and chromatography on Sephadex G-75 and reverse phase (Systen 5) columns of normal and 2-thiouracil-containing tRNAs revealed no profile differences . No accumulation of any precursor tRNA in the thiopyrimidine-treated cells is found . A partial recovery of the pseudouridine content of 2-thiouracil-containing tRNAs can be achieved in vivo by removal of the 2-thiouracil from the culture media . These transfer RNAs have also been used as substrates to study the properties of a partially purified preparation of pseudouridine synthetase II invitro and should be useful as substrates in the further purification of this enzyme.

J Endocrinol, 1977 Sep, 74(3), 405 - 14
Inhibition of thyrotrophin-releasing hormone responsiveness by physiological concentrations of thyroid hormones in the cultured rat pituitary gland; Lewis M et al.; Diced quarter anterior pituitaries from mature females Wistar rats were cultured in synthetic medium with or without added serum . Using each culture as its own control, the thyrotrophin-releasing hormone (TRH) dose-thyrotrophin (TSH) response characteristics of both media were similar; significant TSH secretion being stimulated at TRH doses around 1-5 X 10(-9) mol/l . During days 1-3 of culture, basal TSH secretion fell significantly but TRH responsiveness was unchanged . Neither tri-iodothyronine (T3) nor thyroxine (T4) influenced basal TSH secretion . In both culture media inhibition of TRH responsiveness was demonstrated with concentrations of T3 and T4 within the ranges 1-5 X 10(-12) to 1-5 X 10(-9) mol/l for T3 and 6-5 X 10(-10) to 6-5 X 10(-7) mol/l for T4 . Equivalent inhibition was accompanied by similar T3 concentrations whether T3 or T4 supplements were used, suggesting that T4 itself has no feedback action . The similar concentrations of T3 required to inhibit TRH responsiveness in media either with or without serum suggest that the pituitary is responsive not only to free but also to total thyroid hormone concentrations, since serum-free medium contains no thyroid hormone-binding protein.

Exp Hematol, 1977 Sep, 5(5), 392 - 8
Identification of erythropoietin producing cells in fetal mouse liver cultures; Gruber DF et al.; Fetal mouse liver explants were cultured and the culture media shown to possess an erythropoietically active substance neutralizable by antiserum to sheep plasma erythropoietin, suggesting that the media contain erythropoietin . Immunofluorescent and carbon particle ingestion techniques suggest that the erythropoietin was elaborated by macrophages or Kupffer cells of the hepatic reticuloendothelial system.

Exp Hematol, 1977 Sep, 5(5), 385 - 91
Erythropoietin, thrombopoietin and colony stimulating factor in fetal mouse liver culture media; Zucali JR et al.; Fetal mouse livers, days 13 to 19 of gestation, were cultured for 21 days and the culture media tested for erythropoietin (Ep), thrombopoietin (TSF) and colony stimulating factor (CSF) . High levels of both Ep and CSF were released into the culture media . However, no detectable TSF was found . Maximum Ep culture activity was detected in day 14 and day 15 fetal liver cultures while maximum CSF was found in the day 16 fetal liver culture . These studies indicate that fetal liver cells in culture are capable of producing and/or releasing both Ep and CSF but not TSF.

Biochim Biophys Acta, 1977 Aug 11, 483(2), 493 - 8
The detection and characterisation of collagenase inhibitors from rabbit tissues in culture; Murphy G et al.; As tissue cultures, rabbit bone, skin and non-gravid uterus synthesise inhibitors of collagenase (EC 3.4.24.3) . An assay for the inhibitors is described and their action on collagenase from different tissue sources demonstrated . Evidence for the involvement of the tissue inhibitors of collagenase in the latency of the enzyme in culture media is presented . Latent collagenase was activated by treatment with 4-aminophenylmercuric acetate, and then reacted with the inhibitors to form inactive complexes with properties similar to the naturally occurring latent enzyme forms . The associated changes in molecular weight are detailed, and discussed in relation to the observations of other workers concerning the extracellular control of collagenase activity.

Surgery, 1977 Aug, 82(2), 260 - 5
Erythrocytes cultured from bone marrow; Kelly G et al.; The long-range objective of this study is in vitro tissue culture of bone marrow stem cells to produce erythroid cells of sufficient volume for clinical transfusion . Bone marrow from dogs and patients was cultured in 29 experiments lasting up to 15 weeks . Peripheral erythroid cells from dogs were cultured in three control experiments . Optimal tissue culture media was NCTC 109 augmented with vitamin B12, erythropoietin (EP), folic acid, and 9% fetal protein in 60 mm glass Petri dishes . Additional media and Step III EP was added at 2 to 4 day intervals . Peripheral erythroid cells in culture all were dead within 4 weeks . Marrow erythroic cells in culture proliferated as demonstrated by (1) Fe59 incorporation into cells during culture, (2) H3 thymidine uptake into cultured cells, (3) microscopic evidence of mitoses, and (4) total erythrocyte concentration in cultures far exceeding that of peripheral culture controls . For as yet unexplained reasons the total mature red blood cell concentration in the culture media remained essentially constant throughout the studies . This is a first step in achieving the ultimate goal of bulk erythrocyte production from tissue culture.

J Cell Physiol, 1977 Aug, 92(2), 303 - 7
Decreased in vivo and in vitro colony stimulating activity responses to bacterial lipopolysaccharide in C3H/HeJ mice; Russo M et al.; Mice of the C3H/HeJ strain exhibit low inflammatory and immunological responses to certain bacterial lipopolysaccharide (LPS) preparations . Lymphocytes from C3H/HeJ mice also show defective LPS-induced mitogenesis . We tested the colony stimulating activity (CSA) response of C3H/HeJ mice . As controls we used mice of the congenic C3HeB/FeJ strain, which are good responders to LPS . Serum was obtained three hours after intravenous administration of LPS . Serum CSA was determined in agar cultures of bone marrow cells from AKR mice . The serum CSA response of C3H/HeJ to 10 microgram LPS was approximately 8-fold lower than that of control C3HeB/FeJ mice . In contrast, both strains showed similar serum CSA-induced by Poly I:poly C . Peritoneal macrophage cultures were also incubated with 0.1-10.0 microgram LPS and culture media assayed for CSA . The response of C3H/HeJ macrophages was about 6-fold lower than that of macrophages obtained from the control mice . The results show that the lower responsiveness of C3H/HeJ mice to LPS also extends to the production of CSA . The in vitro findings indicate that the postulated defect in the LPS receptor of B lymphocytes may also be present on macrophages.

Cancer Res, 1977 Aug, 37(8 Pt 2), 2860 - 5
Approaches for the isolation of biologically functional tumor-associated antigens; Reisfeld RA et al.; Melanoma-associated antigens were isolated from human melanoma cells in long-term tissue culture and from the spent culture fluid of these cells propagated in chemically defined, serum-free media . The 3 M KCl extracts from such cells and their concentrated spent culture media elicited specific delayed cutaneous hypersensitivity reactions in patients with malignant melanoma but not in patients with other neoplasms . HLA antigens present in these extracts could be specifically removed by ultracentrifugation in KBr at a density of 1.23 g/ml . Purification of melanoma-associated antigens was achieved by this step, followed by ion-exchange chromatography and preparative isoelectric focusing on Pevikon C870 . Another approach is described for the isolation of carcionembryonic antigens from metastatic lesions with an approximately 70% yield utilizing the least denaturing procedures, which avoid lyophilization and involve essentially 0.9% NaCl solution extraction, specific adsorption, elution from concanavalin A Sepharose, and subsequent gel-exclusion chromatography on Ultrogel AcA 22 . For effective isolation of carcinoembryonic antigens freely shed from cultured cells derived from a primary colon tumor, a system was devised based on the use of Amicon hollow fiber culture units, in which cultured tumor cells were introduced in the extracapiliary spaces of such a unit . The extracapillary fluid, containing carcinoembryonic antigens but no fetal calf serum components, is removed and further purified by affinity chromatography.

J Endocrinol, 1977 Aug, 74(2), 175 - 84
Inhibitory action of porcine follicular fluid upon granulosa cell luteinization in vitro: assay and influence of follicular maturation; Ledwitz-Rigby F et al.; Culture medium 199 supplemented with follicular fluid from 1-2 mm antral porcine follicles inhibited spontaneous luteinization of granulosa cells from preovulatory porcine follicles in vitro . Three characteristics of luteinization were inhibited: morphological transformation, progesterone secretion, and accumulation of cyclic AMP in response to LH . The last was inhibited more effectively by culture media containing 50% follicular fluid than by media containing 20% follicular fluid . The inhibitory actions of the follicular fluid were not altered by charcoal or petroleum ether extraction . Follicular fluid from large follicles (6-12 mm) did not exhibit any of these inhibitory actions . These observations may indicate the presence of a luteinization inhibitor in the fluid of small follicles which (1) is lost by the time the follicle reaches the preovulatory stage, or (2) is overcome by a stimulatory agent which may accumulate as the follicle grows.

Z Parasitenkd, 1977 Jul 29, 52(3), 281 - 8
Immune mechanism of rats on Nippostrongylus brasiliensis in vitro . II . The influence of lymphocytes and peritoneal cells; Horchner F et al.; Nippostrongylus were collected from the intestines of rats 6 days p.i . and kept under sterile conditions in cultures . Serum, lymphocytes and peritoneal cells of immune or non-infected animals were added in various combinations to the culture media . The culture media were changed 2-3 times in an experimental period of 10 days, resp . serum and cells were added . The lymphocytes were isolated from the peripheral blood or from the mesenterial lymph nodes whereas the mononuclear cells were obtained from the peritoneal cavity . Serum and lymphocytes from the peripheral blood both from immune and non-infected rats, had no increased lethal effect on Nippostrongylus . The highest lethality rate of adults (65-68%) was achieved in cultures with peritoneal cells and lymphocytes from the lymph nodes of sensitized rats . Serum of infected or non-infected animals had no influence on adult Nippostrongylus in cultures with these cell combinations . In the controls without any cell-supplements the survival rate of the parasites was up to 88%.

Int J Cancer, 1977 Jul 15, 20(1), 15 - 20
RNA-reverse transcriptase complex from cultured human myeloma-leukemia cells; Sawada H et al.; A high molecular weight RNA-reverse transcriptase complex in the culture media of peripheral leukocytes obtained from two Japanese patients with myeloma-leukemia was detected by demonstration of a 3H-uridine peak and a peak of DNA polymerizing activity banding at a density of 1.15-1.19g/ml . The enzyme in the complex was able to utilize poly(rA)-d(pT)10 or poly (rC)-d(pG) 12-18, but not poly (dA)-d(pT) 10 or (dT) 12-18 as template-primers . The sucrose density sedimentation analysis revealed that RNA in the complex sedimented at a location of approximately 50s and 20-30s.

Acta Physiol Pol, 1977 Jul-Aug, 28(4), 391 - 5
Dependence of vitamin B12 levels on the growth rate of cells cultured in vitro; Koziorowska J et al.; Vitamin B12 content has been determined in different kinds of cells grown in vitro and in the culture media . The results indicate a dependence of the vitamin levels on the growth rate and growth characteristics.

J Immunol, 1977 Jul, 119(1), 227 - 31
Enhancement of weak mixed lymphocyte-type reactions by hydrophilic polymers; Ben-Sasson SA et al.; Two hydrophilic polymers, dextran and polyethylene glycol, were found to enhance in vitro MLR-type reactions which are weak or nonexistent in normal culture media . BALB/c and C57BL/6 thymocytes were cultured for 4 days in Eagle's MEM + 10% FCS, with or without the x-irradiated lymphomas LSTRA AND EL-4, respectively . By thymidine incorporation no stimulation was observed with EL-4 in plain culture whereas occasionally a very small stimulation was observed with LSTRA . Four percent dextran (w/v, m.w . greater than or equal to 40,000) improves stimulation (E/C) by about 2-fold . The effect of polyethyleneglycol m.w . 6,000 (PEG-6) is much more pronounced . At the optimal concentration of 4 to 5%, PEG-6 causes an increase of 5- to 20-fold in the stimulation index . The PEG-6 effect can also be observed microscopically by the appearance of numerous blasts clustered around the tumour cells . PEG-6 by itself has little effect on thymocytes in the absence of tumor cells . In the case of one-way allogeneic MLR under suboptimal ratios, dextrans (m.w . greater than or equal to 40,000) enhance the reaction dramatically . We speculate that these effects are occurring via a change in the solvent (such as solvent exclusion) in a way which enhances immunologic interactions under conditions which are tolerable to cells in vitro.

Invest Urol, 1977 Jul, 15(1), 2 - 4
Synthesis of alpha-fetoprotein and some other serum proteins in testicular tumors; Sakashita S et al.; Serum levels of alpha-fetoprotein were determined in 33 patients with testicular germ cell tumors and were normal in 11 patients with seminoma and in one patient with matured teratoma; high levels were observed in 19 of 21 patients with embryonal carcinoma, teratocarcinoma, or a mixed type of these germ cell tumors . Tissues from the testicular germ cell tumors were cultivated with 14C-labeled leucine . After incubation, the culture media were subjected to immunoelectrophoretic and autoradiographic analyses . The results were: (i) Radioactive alpha-fetoprotein, albumin, transferrin, and alpha1-globulin appeared in the culture media of embryonal carcinomas obtained from two infants . (ii) Radioactive albumin and alpha1-globulin appeared in the culture media of a mixed type tumor metastasized from testis to retroperitoneal region . (iii) No such radioactive proteins appeared in the culture media of primary seminomas.

J Endocrinol, 1977 Jun, 73(3), 511 - 7
Analysis of androgen-binding protein in media from sertoli cell incubations and cytosols from rat testes; Rommerts FF et al.; Binding ability of androgen-binding protein (ABP) in concentrated media from Setoli cell cultures and in various preparations of cytosols from rat testicular tissue have been estimated using dialysis and polyacrylamide gel electrophoresis (PAGE) . Equilibrium dialysis in the presence of 1 nM-17beta-hydroxy-5alpha-androstan-3-one appears to be the most convenient and reliable method for estimation of ABP if no other binding proteins are present . PAGE can be applied for estimation of ABP in samples which also contain other steroid-binding proteins . Estimated binding activities in culture media measured by PAGE and dialysis were essentially similar . Binding of androgens to ABP estimated with PAGE in cytosols prepared after sonication of testicular tissue was much lower than in cytosols obtained after homogenization using a Potter-Elvehjem homogenizer . When ABP was added to cytosols prepared after sonication or homogenization, approximately 50 and 90% respectively of the original binding of androgens to ABP was recovered.

Scand J Clin Lab Invest, 1977 Jun, 37(4), 351 - 6
Identification of glutamine as a hepatic factor which influence the synthesis of collagen by freshly isolated fibroblasts; Ronnemaa T et al.; Earlier we have found that 35,000 g supernatants of liver homogenates from both normal and hypercholesterolaemic rats stimulate the synthesis of collagen by freshly isolated fibroblasts . Supernatants from the fatty livers of hypercholesterolaemic rats showed greater stimulation . In the present study we fractionated the 35,000 g supernatants using gel filtration, ion-exchange chromatography, paper electrophoresis and paper chromatography . One of the stimulating factors turned out to be glutamine . However, the concentration of glutamine was the same in normal and fatty livers suggesting that glutamine is not responsible for the greater stimulating activity found in the 35,000 g supernatants from fatty livers . Authentic glutamine increased the synthesis of collagen by 40% at a concentration of 40 mumol/l but inhibited it 80% at 4 mmol/l, which is widely used in cell culture media . There, the concentration of glutamine should be controlled carefully in tests for collagen synthesis in vitro.

J Parasitol, 1977 Jun, 63(3), 418 - 26
Decreased binding of cytotoxic antibody by developing Schistosoma mansoni . Evidence for a surface change independent of host antigen adsorption and membrane turnover; Dean DA; Schistosoma mansoni schistosomula maintained in chemically defined culture media became increasingly resistant to the cytotoxic effects of infected guinea pig serum . Two- and 6-day-old schistosomula recovered from mice showed no uptake of IgG antibody from infected guinea pig serum, as revealed by the indirect fluorescent antibody test . Results from this test remained negative when the schistosomula were tested at 0 C, after exposure to drugs which inhibit synthetic and secretory processes, or after being killed by heat or formalin . In contrast, new schistosomula collected within 3 hr after skin penetration bound IgG from infection serum under all test conditions, and showed increased susceptibility to cytotoxicity after exposure to various drugs . It thus appears that soon after skin penetration schistosomula undergo surface changes which prevent binding of antibody from infection serum . These changes can apparently take place in the absence of host antigens, and once they have occurred, do not depend on worm physiological processes for their function.

Gann, 1977 Jun, 68(3), 363 - 7
Production of alpha-fetoprotein by cultured rat ascites hepatoma cells determined in vivo as alpha-fetoprotein-negative; Isaka H et al.; Three ascites hepatoma cells of the rat, AH-41B, AH-34, and AH-64B, which had been determined in vivo as alpha-fetoprotein-negative, were cultivated in vitro in a medium containing adenosine 3',5'-cyclic monophosphate (cyclic AMP), cyclic dibutyryl-AMP, or theophylline . The concentration of alpha-fetoprotein in culture media was measured by the 125 I-radioimmunoassay . Results demonstrated that the AH-41B cells produced alpha-fetoprotein in vitor, the concentration of which being elevated in the media with three substances, while the remaining AH-34 and AH-64B cells did not . A comment was made on the producibility of alpha-fetoprotein in the so-called alpha-fetoprotein-negative hepatoma cells.

Am J Obstet Gynecol, 1977 May 15, 128(2), 209 - 11
Electron probe microanalysis of the chemical elemental content of human follicular fluid; Chong AP et al.; Follicular fluid samples were obtained by puncturing follicles of ovaries in situ from patients undergoing laparotomy . Sodium, potassium, chloride, magnesium, calcium, phosphorus, and sulfur concentrations measured by electron probe microanalysis were similar to those of blood, with minimal differences . This suggested that culture media in which these electrolytes are added in concentrations similar to those of serum are appropriate for culture of the human oocyte.

Experientia, 1977 May 15, 33(5), 668 - 70
Degradation of {3H}thymidine by a pentosyltransferase (EC 2.4.2.4) in the plasma of man and different animals; Pauly JL et al.; {3H}Thymidine is degraded by an enzyme (thymidine phosphorylase; EC 2.4.2.4) which we have identified in the plasma of man and some animals . The presence of this enzyme in plasma or sera used to supplement culture media may, under certain experimental conditions, limit the validity of measuring the uptake of radiolabeled thymidine as a means of defining DNA synthesis.

In Vitro, 1977 May, 13(5), 293 - 6
Effect of activated charcoal on callus growth and shoot organogenesis in tobacco; Constantin MJ et al.; Incorporating activated charcoal (AC) in culture media has been shown to affect growth and development of various organisms . Since AC stimulates the development of tobacco haploid plantlets from cultured anthers, research was conducted to determine the effect of activated charcoal on pith-derived callus growth and shoot development in Nicotiana tabacum cv . Wisconsin 38 . Our results indicate that the hormones required for callues growth and shoot development in Wisconsin-38 tobacco are adsorbed by AC, thereby inhibiting callus growth and prohibiting shoot development . This effect was observed even when AC was removed from the medium by filtration prior to culturing the callus.

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