Culture medium

Invest Ophthalmol Vis Sci, 1977 May, 16(5), 392 - 6
Aldose reductase in retinal mural cells; Buzney SM et al.; Cultured mural cells (intramural pericytes) from adult rhesus retinal capillaries were examined for the presence of the sorbitol pathway . A radioimmunoassay for human aldose reductase, cross-reactive with rhesus lens aldose reductase, showed the presence of this enzyme in our cultured cells . Mural cells grown in culture media containing normal (10 mM) and high (40 mM) levels of glucose were examined for polyol accumulation by gas-liquid chromatography . Cells incubated in high glucose medium showed a threefold increase in sorbitol concentration over cells grown at low glucose levels . After 30 days in high glucose medium, mural cells formed dense multilayered areas with extensive cellular debris . These findings suggest the presence of the sorbitol pathway in cultured retinal mural cells and cellular degeneration in high glucose medium; this may have possible implications in the pathogenesis of diabetic retinopathy.

Biochem J, 1977 May 1, 163(2), 303 - 7
Evidence that latent collagenases are enzyme-inhibitor complexes; Sellers A et al.; Specific collagenase from the culture media of various rabbit tissues and cells exists in active and latent forms . Latent collagenase is most effectively activated with 4-aminophenylmercuric acetate, a thiol-blocking reagent, strongly suggesting that latent forms are enzyme-inhibitor complexes . A collagenase inhibitor from bone cultures, which may be closely related to the inhibitor of such latent enzyme complexes, was partially characterized.

Schweiz Med Wochenschr, 1977 Apr 16, 107(15), 514 - 20
{Therapy of hepatitis B . Practical implications}; Kuhn HA; There is as yet no specific treatment for viral hepatitis, and in an uncomplicated course no further action apart from moderate bedrest is necessary . The patient should however, be isolated in a special ward . In fulminant hepatic failure the benefit of glucocorticoid therapy is still controverted and appears to depend on an early beginning . Treatment with HBsAg-rich human serum in fulminant hepatitis B is still under evaluation . In chronic active hepatitis the administration of azathioprine in combination with glucocorticoids is highly effective, and it appears to be irrelevant whether HBsAg is present in plasma or not; however, the best results have been achieved in "lupoid" hepatitis . The use of transfer factor, laevamisol or thymosin to suppress T-cell action on antibody production of the B-cells cannot yet be finally evaluated with respect to its effectiveness in chronic active hepatitis . Prevention is of major importance in solving the problems involved in hepatitis B infections . Recent experience with active immunization using HBsAg-rich sera or purified formalin inactivated HBsAg preparations suggest the possibility of successful vaccination against hepatitis B in the near future, but the precondition for obtaining sufficient quantities of vaccine is to find suitable culture media for the virus.

Ann Immunol (Paris), 1977 Apr-Jun, 128C(3), 645 - 51
Stimulation of human lymphocytes in vitro by purified autologous DNA; Kirsch-Volders M; Addition of increasing concentrations of autologous DNA, but not of allogeneic DNA, in culture media of human lymphocytes induces a parallel increase of thymidine incorporation into leucocytes and of lymphocytes transformation into blast cells . Specificity of DNA action was analysed by different DNase and RNase treatments . DNase or RNase alone neither stimulates blastic transformation of lymphocytes and incorporation of thymidine into leucocytes, nor inhibits PHA-induced stimulations of these cells . However Dnase--but almost not RNase--clearly inhibits thymidine incorporation and blast transformation induced by autologous DNA.

Can J Microbiol, 1977 Apr, 23(4), 363 - 8
Inhibition of gas vesicle production in Microcyclus aquaticus by L-lysine; Konopka AE; The timing and degree of gas vesicle production in Microcyclus aquaticus was affected by nutritional conditions . If 50 microng L-lysine/ml was added to a glucose-mineral salts medium (DM), the organism did not form gas vesicles . This effect was specific for L-lysine, as neither D-lysine nor meso-diaminopimelic acid prevented gas vesicle production . Cells grown in the presence of L-lysine did not contain any immunologically detectable gas vesicle protein, which indicates that L-lysine affects expression of the structural gene for the gas vesicle protein rather than assembly of the protein into gas vesicles . The addition of L-lysine to cultures in DM did not immediately decrease the rate of gas vesicle assembly, nor did the removal of cells from DM plus L-lysine to DM result in immediate gas vesicle production . Gas vesicle production was also affected by the addition of L-threonine or L-cysteine to culture media or by an increase in the medium's ionic strength . These results are discussed in relation to the aspartic acid pathway of amino acid biosynthesis and effects upon the intracellular L-lysine concentration.

Am J Med Technol, 1977 Apr, 43(4), 345 - 8
Mycological culture media and its quality control; Otto V; The need for control of the quality of media used in the isolation and identification of fungi is an important aspect of the overall quality control of laboratory culture media; however, guidelines for such controls have not been available . In this paper, primary isolation media with and without appropriate drugs are suggested for use on a variety of specimens . Positive and negative quality control organisms that indicate clear-cut postiive and negative results on the fungal media to be tested are listed . Several alternative procedures for maintenance of fungi are discussed.

J Clin Endocrinol Metab, 1977 Apr, 44(4), 651 - 9
Effects of fibroblast growth factor and epidermal growth factor on the rate of growth of amniotic fluid-derived cells; Gospodarowicz D et al.; Primary cultures of cells derived from human and bovine amniotic fluid were cultivated in Dulbecco's modified Eagles medium with 20% fetal calf serum . Whereas increased concentrations of fetal calf or other types of serum bring about no further mitogenic response, the rate of growth of these cells was nevertheless sharply increased when 100 ng/ml of FGF was added to the culture media . The FGF induced mitogenic effect was statistically significant . EGF at 100 ng/ml had a less pronounced effect which, when analyzed statistically, was not significant . Amniotic fluid-derived cells are used for the prenatal detection of genetic disorders . The use of FGF to increase the rate of growth of these cells should reduce the time required between amniocentesis and diagnosis.

Exp Hematol, 1977 Mar, 5(2), 103 - 8
Stimulation of erythropoiesis in a grafted animal by mouse fetal liver culture media; Zucali JR et al.; Erythropoietin given to polycythemic, irradiated mice receiving a bone marrow transplant restores the number of erythroid colonies produced . In this study mouse fetal liver culture media administered to polycythemic, irradiated mice also restored the number of erythroid colonies and erythropoiesis as measured by 59Fe incorporation . Thus, mouse fetal liver in culture produces a factor which resembles erythropoietin.

Am Rev Respir Dis, 1977 Feb, 115(2), 285 - 93
Cell line A549: a model system for the study of alveolar type II cell function; Smith BT; Cell line A549, a continuously cultured line derived from a human pulmonary adenocarcinoma that has morphologic and biochemical features of the pulmonary alveolar type II cell, was studied with regard to saturated phosphatidylcholine synthesis and secretion . Evaluation of a number of culture media showed that those media that supported the most rapid rate of in vitro growth were least optimal for saturated phosphatidylcholine synthesis . Similarly, a labeled precursor (choline labeled with tritium) was most efficiently converted to saturated phosphatidylcholine when growth was slow or arrested, whereas during phases of active cellular proliferation a greater proportion was incorporated into unsaturated (structural) phosphatidylcholine . De novo synthesis of phosphatidylcholine was primarily via the cytidine diphosphate-choline pathway, because no incorporation of labeled methyl groups from S-adenosyl-L-methionine was observed . Cortisol stimulated tritium-labeled choline incorporation into saturated phosphatidylcholine in a manner that likely involed a steroid receptor system, whereas cortisone was inactive . The half-life of cellular saturated phosphatidylcholine was of the order of 100 hours and could be accounted for virtually entirely by release of this phospholipid into the medium unchanged . Both cholinergic and adrenergic agents in high doses modestly stimulated the release of pre-labeled saturated phosphatidylcholine from the cells, and the response to these secretagogues was significantly enhanced by cortisol . Cell line A 549 promises to be a useful model system for studies of the cellular events involved in pulmonary saturated phosphatidylcholine synthesis and secretion.

J Natl Cancer Inst, 1977 Feb, 58(2), 215 - 21
Enzymatic susceptibility and spontaneous release of human melanoma tumor-associated antigens; Stuhlmiller GM et al.; A chimpanzee anti-human melanoma antiserum was used to study the enzymatic susceptibility and spontaneous release into tissue culture medium of human melanoma tumor-associated antigens (TAA) . Limited proteolytic digestion of melanoma cells with trypsin or with pronase rendered these cells refractory to lysis by the chimpanzee antiserum and complement . Longer periods of incubation of higher concentrations of enzyme caused an increased sensitivity to lysis . Digestion of melanoma cells with neuraminidase apparently exposed antigens reactive with natural antibodies in rabbit complement because cells so treated had a marked increase in sensitivity to cytolysis . Absorption of the complement with either neuraminidase-treated human melanoma cells or washed human spleen cells prior to its use in the cytotoxicity assay removed this activity . When absorbed complement was used, neuraminidase had no noticeable effect on the expression of malanoma TAA . These results suggest that proteolytic digestion of melanoma cells may prove to be a useful means of solubilizing TAA . The spontaneous release of melanoma cell membrane TAA was studied . Protein precipitated by (NH4)2SO4 from four of six samples of tissue culture medium used to feed malanoma cell lines contained significant antigenic activity compared to a control "antigen" preparation, whereas one preparation contained only limited TAA activity . One melanoma cell line that apparently failed to release TAA into the culture medium had previously become nonreactive with the chimpanzee antiserum . From these data, we conclude that melanoma cells growing in tissue culture rapidly release large amounts of TAA into the culture media and, as a result, the spent culture medium may be a good source for obtaining TAA for further study . The significance of these results is discussed.

J Chromatogr, 1977 Jan 21, 131, 383 - 90
Rapid purification of an RNA tumor virus and proteins by high-performance steric exclusion chromatography on porous glass bead columns; Darling T et al.; High-performance steric exclusion chromatography on a 1250-A pore size polyethylene glycol-treated glass bead column was used to purify avian myeloblastosis virus and hamster melanoma virus from plasma protein and tissue culture media . The purified hamster melanoma virus was still infectious and the avian myeloblastosis virus-associated RNA-directed DNA polymerase showed a 1100-fold purification of the virus from one column treatment . Electron microscopy of the purified virus showed intact particles, with surface projections evident . The time required for column purification of the virus was 5 min.

Biochim Biophys Acta, 1977 Jan 11, 480(1), 178 - 93
Control of the production and partial characterization of repressible extracellular 5'-nucleotidase and alkaline phosphatase in Neurospora crass; Hasunuma K et al.; A new species of orthophosphate repressible extracellular 5'-nucleotidase (5'-ribonucleotide phosphohydrolase, EC 3.1.3.5) was found to be released into mycelial culture media when a wild type strain of Neurospora crassa was grown on limiting amounts of phosphate . The production of 5'-nucleotidase and extracellular acid and alkaline phosphatase was inhibited by the addition of rifampicin when it was added at the later stage of mycelial growth, but not when it was added at a very early stage . The 5'-nucleotidase and extracellular alkaline phosphatase were partially purified and characterized . pH optimum of the former was 6.8 and that of the latter was higher than 10.0 . The 5'-nucleotidase activity was inhibited by ethylenediaminetetraacetate (EDTA) and ZnCl2 at pH 6.8 and stimulated by MnCl2 and CoCl2 at pH 4.0 . Alkaline phosphatase activity was stimulated by EDTA, MgCl2, CoCl2 and MnCl2 . 5'-nucleotidase activity was stimulated by EDTA, MgCl2, CoCl2 and MnCl2 . 5'-nucleotidase hydrolyzed various 5'-nucletides but not 3'-nucleotides or other various phosphomono- and diester compounds . Alkaline phosphatase hydrolyzed all the phosphomonoester compounds tested . Mutants, nuc-1 and nuc-2, which were originally isolated by the inability to utilize RNA or DNA as a sole source of phosphate, were unable to produce 5'-nucleotidase or six other repressible enzymes reported previously . These mutants showed no or significantly reduced growth on orthophosphate-free nucleotide media depending on the number of conidia inoculated, mainly because of loss of ability to produce these repressible extracellular phosphatases.

Endocrinology, 1977 Jan, 100(1), 39 - 45
The culture of pig oocytes in minimal medium, and the influence of progesterone and estradiol-17beta on meiotic maturation; McGaughey RW; Pig oocytes, with (intact) or without (denuded) adhering granulosa cells, were cultured for 33 to 35 h in minimal culture media containing bovine serum albumin and either progesterone or estradiol-17beta or both steroids . The incidence of maturation for intact oocytes in minimal medium was comparable, if not superior, to that observed by other investigators using complex culture media . No statistically significant stimulation of maturation by progesterone was observed for either intact or denuded oocytes . Estradiol-17beta at concentrations of 1.0 and 10.0 mug per ml inhibited the maturation of denuded oocytes, but the inhibition was overcome either by including progesterone in the medium or by subsequently culturing the inhibited oocytes in steriod-free medium . Chromosomal examination of air-dried oocytes indicated that intact and denuded oocytes cultured either in progesterone or estradiol-17beta exhibited an increased incidence of diploidy and hyperhaploidy at telophase I and metaphase II, but that oocytes cultured in media containing both progesterone and estradiol showed a higher incidence of normal haploidy than did control oocytes.

Endocrinology, 1977 Jan, 100(1), 205 - 8
Chromatographic evidence for vasotocin biosynthesis by cultured pineal ependymal cells from rat fetuses; Pavel S et al.; Cultured pineal ependymal cells from rat fetuses aged 17 to 19 days post-coitum release into their media a substance that has hydroosmotic, antidiuretic and rat uterine activities . Paper chromatography of concentrates from culture media demonstrates that the substance possessing all the above activities was eluted from a slow moving region (Rf 0.15-0.35) with a peak at 0.20-0.30 . This region is not significantly different from the Rf of synthetic arginine vasotocin (AVT) used as standard . Neither hydroosmotic nor rat uterine activities could be detected in the chromatographic eluates from the fast moving region corresponding to the Rf of synthetic oxytocin (0.55-0.65) used as standard . The chromatographic mobility of biological activities from culture media, the ratio of the activities as well as their susceptibility to tryptic digestion, demonstrates the presence of a basic peptide indistinguishable from synthetic AVT . The total amount of AVT released into the medium during 43 days of incubation is about 40 times greater than the amount contained in non-incubated pineal glands of the same age, strongly suggesting de novo synthesis of AVT.

Curr Med Res Opin, 1977-78, 5(4), 315 - 27
Experimental in vitro and in vivo comparison of modern antimycotics; Haller I et al.; Studies were carried out with the current leading antimycotic agents, using identical test methods, to compare their activity in vitro and in animal in vivo experiments . In the broth dilution test, the minimum inhibitory concentrations against approximately 50 strains of the most important species of yeasts, dermatophytes and moulds were determined using different culture media . Efficacy against yeasts was examined in experimentally-induced candidosis in mice treated orally, and against dermatophytes in experimentally-induced trichophytia in guinea pigs receiving topical treatment . The results showed that the imidazole derivatives studied can be regarded as true broad-spectrum antimycotics, and clotrimazole, in particular, had a well-balanced overall effect . The significance of the experimental data with respect to therapeutic efficacy in man in various indications is discussed.

Dermatologica, 1977, 155(4), 224 - 8
{Trial of the culture media Microcult GC for detection of gonococci}; Olmos L et al.; A trial of the culture medium 'Microcult GC' for the detection of gonorrhea in 246 patients confirms its utility . The direct smear of the exudate is still indispensable . The best culture medium is the Thayer-Martin plate . The main advantage of the new medium tested is its simple utilization technique which makes it suitable for the practitioner.

Arch Environ Contam Toxicol, 1977, 5(4), 437 - 45
The effect of 2,4-dichlorophenoxyacetic acid on growth and nitrogen-fixation of blue-green alga Anabaenopsis raciborskii; Das B et al.; Sodium salt of 2,4-dichlorophenoxyacetic acid (80% active ingredient), commonly applied for the control of aquatic weeds, was used to observe its effect on the growth and nitrogen fixation of a heterocystous bloom forming blue-green alga Anabaenopsis raciborskii . A concentration of 10 microgram per ml of 2,4-D showed stimulation of growth and nitrogen fixation and these were almost unaffected in presence of its 100 microgram per ml in the medium . The alga could tolerate up to 800 microgram per ml in liquid culture media with and without nitrate nitrogen and up to 90 microgram per ml on to agar plates . Nitrogen fixation was inhibited in presence of its higher concentrations.

Endocrinology, 1977 Jan, 100(1), 216 - 26
Induction of hypothyroidism and hypoprolactinemia by growth hormone producing rat pituitary tumors; Seo H et al.; The GH3 rat pituitary tumor cell line which secretes both growth hormone (GH) and prolactin (PRL) stopped releasing PRL when transplanted to animals; furthermore, it suppressed PRL production by the hosts' pituitary glands . When the same tumor was transferred back to cell culture, PRL production resumed . The PRL to GH ratio in cell culture medium and cells ranged from 5 to 1 while in the tumor and serum of the host animals it averaged 0.09 and 0.001, respectively . To investigate further this phenomenon, female rats were transplanted with GH3 tumors (T) and compared to intact normal (N) and to thyroidectomized (Tx) rats . T animals were larger and had splanchnomegaly but smaller pituitaries and thyroids . Serum PRL concentrations in the basal state were decreased, as were levels of triiodothyronine (T3), thyroxine (T4), and free T4 index . Despite reduced serum thyroid hormone concentrations, and in contrast to Tx animals, the serum thyrotropin (TSH) level in T rats was not elevated and they did not show a supranormal TSH response to thyrotropin-releasing hormone (TRH) administration . The PRL response to TRH in T animals was completely abolished while all N and Tx animals responded by a significant increase in serum PRL . Serum corticosteroids and estrogens were normal in T rats . Pituitary content of PRL was decreased and that of TSH increased in T rats . Tx animals, however, had a reduced pituitary content of PRL, TSH, and GH . When GH3 cells were grown in cell culture media containing serum from T animals, there was a reduction of PRL content in cells and released in the medium . Addition of T3 to the T serum did not alter its suppressive effect on PRL nor did rat GH added to N serum alter PRL production and release in vitro . In a preliminary experiment, rats injected ip with 50 mug hGH in two divided doses for eighteen days, suppressed serum T4 and T3 concentrations; pituitary content of TSH was significantly increased and that of PRL slightly decreased . Injection with 250 mug oPRL or saline, on the same schedule and for the same length of time, had no significant effect on the levels of serum thyroid hormones . Thus, GH, but also possibly other substance(s) secreted by GH3 tumors in vivo a) suppress the production of tumor and pituitary PRL; b) suppress the release of TSH, causing mild hypothyroidism; c) inhibit the PRL and TSH responses to TRH; and d) decrease the production of PRL in tissue culture . Although no simple and unifying theory could explain these findings, an hypothesis implicating somatomedin is presented.

Acta Microbiol Acad Sci Hung, 1977, 24(3), 175 - 80
Quantitation of seven elements in Mycobacterium phlei and Mycobacterium bovis by neutron activation analysis; Karasseva V et al.; Quantitative determination of the elements potassium, sodium, manganese, magnesium, iron, cobalt and zinc was performed in mycobacteria by neutron activation analysis . Mycobacterium phlei ATCC 19 249 at different phase of growth (4, 8, 13, 23 and 37 days old cultures), and 14 days old Mycobacterium bovis BCG cultures and uninoculated semi-synthetic Sauton culture media were examined . The elements studied could be divided into three groups; sodium, potassium and magnesium could be regarded as major, iron as minor, and zinc, manganese and cobalt as trace elements . M . phlei contained, with the exception of zinc, higher amounts of elements than M . bovis . Other metals (aluminium, antimony, rubidium) could also be detected.

Adv Chromatogr, 1977, 15, 1 - 31
Detection of bacterial metabolites in spent culture media and body fluids by electron capture gas-liquid chromatography; Brooks JB; Electron capture gas-liquid chromatography, when used to analyze derivatized extracts of spent culture media and body fluids under specified conditions, holds promise as a tool for use by physicians, hospitals, and clinical laboratories in identifying certain diseases and disease-producing organisms . The detection of certain disease processes and the identification of disease-producing organisms are based on qualitative or large quantitative differences in EC-GLC profiles or a combination of both . Various practical procedures are given for extracting and derivatizing compounds, such as carboxylic acids, hydroxy acids, alcohols, amines, and nitrosamines . The characteristics of the parameters essential for successful analysis are discussed . Species and, in some cases, strains have been differentiated by comparing EC-GLC profiles . Metabolic products are affected by change in substrate . Media that can be reproduced from lot to lot are essential in some studies . The volatile components detected by EC-GLC in spent culture media consist mostly of bacterial metabolites, but the volatile compounds detected in body fluids may be bacterial metabolites, volatile components produced by the host in response to an infection, metabolites of cells associated with host defense, or a combination of two or more of these groups of compounds . The EC-GLC profiles obtained by analysis of synovial and cerebrospinal fluids appear to have good potential for use in diagnosing certain forms of arthritis and meningitis . Well-documented samples are essential to establishing EC-GLC profiles representative of a particular disease . A moderately priced computer would greatly aid in data processing and could be especially useful in compensating for minor changes in the retention times of peaks, which can occur as a result of column aging or when columns are renewed . An approach to the identification of components detected by EC-GLC, which makes use of electron capture gas chromatography-mass spectrometry, is presented.

Scand J Immunol, 1977, 6(11), 1133 - 44
Poly A:U-induced secretion of T-lymphocyte helper factors; Bick PH et al.; In vitro exposure of mouse thymocytes to complexes of polyadenylic:polyuridylic acid (poly A:U) effected, within 6 h, the release of soluble factor(s) capable of nonspecifically enhancing IgM and IgG plaque-forming cells (PFCs) in in vitro primary and secondary spleen cell responses to burro erythrocytes . Poly A:U stimulation was, most likely, polyclonal, since production of soluble factor(s) occurred in the absence of antigen and in serum-free culture media . Poly A:U-induced soluble factor(s) were not capable of substituting for T cells but were dependent on T cells for the expression of PFC enhancement . These data support the hypothesis that the mechanism of poly A:U's adjuvant action is polyclonal stimulation of T cells, causing early induction and release of nonspecific, soluble PFC-enhancing factor(s).

J Supramol Struct, 1977, 7(3-4), 515 - 30
Glycoprotein synthesis as a function of epithelial cell arrangement: biosynthesis and release of glycoproteins by human breast and prostate cells in organ culture; Tokes ZA et al.; We demonstrate that a technique is available to investigate glycoprotein synthesis in organ cultures of human breast and prostate surgical specimens where the 3-dimensional epithelial cell arrangement remains intact . Malignant breast and prostate epithelium maintained their capacity to synthesize glycoproteins for at least 3 days as followed by the incorporation of {3H}glucosamine into macromolecules . Over 70% of incorporation was by malignant cells as judged by autoradiography . Labeled glycoproteins were released into glandular lumina and consequently into the culture fluid . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed predominantly one group of macrmolecules released with an apparent molecular weight of 48,000 +/- 6,000 daltons . This glycoprotein was found in all of the breast specimens studied, which included 1 medullary, 1 infiltrating lobular, and 8 infiltrating duct carcinomas . The pattern was independent of the availability of estrogen receptors . A similar glycoprotein was also observed in the culture media from a Grade I and a Grade II well-differentiated infiltrating prostate carcinoma . Incorporation was below the level of detection in 4 of 6 cases of benign prostatic hyperplasia . A more complex pattern of labeled glycoproteins was found in the media of a Grade II and a Grade III poorly-differentiated prostate carcinoma . The established human mammary carcinoma cell line MCF-7 synthesized and released a similar 48,000 molecular weight glycoprotein but additional components with larger molecular weights were also released . An intriguing interpretation that 3-dimensional tissue integrity restricts some glycoprotein synthesis is discussed . Cells grown in 2-dimensional monolayers could escape from such a topographic restriction and express additional families of glycoproteins.

Arch Invest Med (Mex), 1977, 8(3), 199 - 208
Qualitative x-ray spectrometric study of E . histolytica trophozoite nuclei; Feria-Velasco A et al.; Qualitative X-ray energy spectroscopical analysis was performed in two strains of Entamoeba histolytica trophozoites: HK-9; NIH strain, and HM-2; IMSS strain, cultured in axenic and monoxenic conditions, respectively . The trophozoites were fixed in buffered-glutaraldehyde, embedded in Epon, thick sectioned )50-200 nm), and mounted on nickel grids . X-ray emissions were analyzed with an Ortec X-ray energy detector and multichannel and analyzer adapted to a transmission electron microscope . The study was concentrated on the refractive intranuclear bodies, characteristically numerous in axenically grown E . histolytica trophozoites . Nuclei of trophozoites with those intranuclear bodies contained phosphorus, calcium, and magnesium in greater amounts than nuclei not containing those structures . This was true for all axenically grown trophozoites and two monoxenically grown trophozoites which contained four intranuclear bodies . It is concluded that the composition of the refractive intranuclear bodies in E . histolytica trophozites does not appear to change with the culture conditions or with the composition of the culture media.

Microbios, 1977, 19(77-78), 191 - 204
Antigenic variability of Aspergillus fumigatus strains; Kurup VP et al.; The effect of culture media, temperature of incubation, and continuous shaking of cultures, on the reactivity and yield of antigens of Aspergillus fumigatus were evaluated . It was found that AOAC medium was superior to Czapek medium and shake cultures yielded better results than stationary cultures . However, antigens from stationary cultures in AOAC medium incubated at 30 degrees C for 3 weeks were equally as good as antigens obtained from 2-week-old shake cultures . Antigens from 11 selected strains of Aspergillus fumigatus were used to test antibody activity in 33 sera from patients with various forms of aspergillosis and 35 normal controls by the agar gel double-diffusion method . The results showed that the reactivity of individual antigens varied from 42 to 87%, indicating that antigens from more than one strain of Aspergillus fumigatus may be used . The cross-reactivity between strains were studied by two-dimensional immunoelectrophoresis . The use of polyacrylamide gel electrophoresis and crossed immunoelectrophoresis in the quality assurance of Aspergillus antigens is discussed.

Acta Microbiol Pol, 1977, 26(3), 273 - 9
Synthesis of cellulase by Fusarium sp . in different culture conditions; Targonski Z et al.; The mechanism of the synthesis of cellulases by Fusarium sp . strain was studied . It was found that a significant role in determination of cellulases is played by the adsorption of these enzymes on cellulose . The aeration level of the culture media had a significant effect on the synthesis as well as on the enzymatic complexity of cellulases.

Dev Biol Stand, 1976 Dec 13-15, 37, 185 - 90
Effect of bacterial toxins in serum on the chromosomes of WI-38; Whitaker AM et al.; Two batches of foetal calf serum, free from detectable bacterial, mycotic, mycoplasmal and bovine viral conta mination and possessing good growth-promoting properties induced an abnormally high number of chromatid breaks in WI-38 cells to the extent that the cells were unacceptable as a vaccine substrate . This phenomenon, immediately reversible on changing to a different batch of serum, was unaffected by pasteurisation, suggesting that it was caused by some unidentified toxic factor(s) . It was found that the same effect could be brought about by the addition of subcytotoxic concentrations of some bacterial toxins to the culture media . These findings re-emphasise the importance of improved methods for the collection and the maintenance of sterility throughout the processing of such sera, especially when they are intended to be incorporated in media used to establish vaccine substrates.

J Immunol, 1976 Dec, 117(6), 2220 - 5
Subpopulations of chicken B lymphocytes; Lifter J et al.; Immunoglobulin-synthesizing cells from the spleen and bursa were fractionated by the 1 X G sedimentation velocity technique and characterized by their ability to synthesize immunoglobulin and by staining with fluorescent anti-light chain chain . Four subpopulations of immunoglobulin-synthesizing cells were identified . In the bursa, slowly sedimenting (S 2.3 mm/hr) and rapidly sedimenting (S greater than 3.5 mm/hr) subpopulations with surface immunoglobulin were present; in the spleen, a slowly sedimenting (S 2.3 mm/hr) subpopulation with surface immunoglobulin and plasma cells (S greater than 3.5 mm/hr) with large concentrations of intracellular immunoglobulin existed . The subpopulations differed most markedly in their ability to synthesize immunoglobulin (cpm Ig synthesized/10(6) Ig-positive cells); the rates of immunoglobulin synthesis were in the ratio 1:2:1:900 . The slowly sedimenting B cells from the spleen and both subpopulations of B cells from the bursa released small amounts of immunoglobulin into the culture media, whereas, the plasma cells released immunoglobulin at a rate as much as 3700 times greater . Bursal B cells could be further distinguished from splenic B cells by a greater amount of DNA synthesis.

Clin Chim Acta, 1976 Nov 15, 73(1), 157 - 62
Studies on the use of skin fibroblasts for the measurement of cystathionine synthase activity with respect to homocystinuria; Griffiths R et al.; The levels of cystathionine synthase have been examined in cultured skin fibroblasts obtained from a patient suffering from pyridoxine-non-responsive homocystinuria and compared with the normal and heterozygous states . Levels of synthase activity were found to vary with time in culture and composition of culture media . The physical properties of normal and abnormal synthase activities were markedly different . It has been concluded that caution has to be exercised when using cell culture methods for the purpose of defining normal, heterozygous and homozygous states with reference to homocystinuria.

Scand J Dent Res, 1976 Nov, 84(6), 353 - 6
Differentiation of odontogenic tissues in organ culture; Thesleff I; Molar tooth germs from 17-d-old mouse embryos were cultivated in a Trowell-type culture, and different culture media were tested for their ability to support enamel formation . The medium which allowed secretion of considerable amounts of enamel matrix by ameloblasts consisted of BGJb medium supplemented with 20% horse serum, 10% chick embryo extract and 0.9 mM ascorbic acid . At the onset of culture the teeth were in the early bell stage . After 2 weeks of cultivation both odontoblasts and ameloblasts had differentiated, and considerable amounts of predentin and enamel matrix had been secreted . Similar development was also seen in teeth which had been enzymatically separated into the mesenchymal dental papilla and epithelial enamal organ and subsequently recombined in vitro . This method allows good differentiation of odontogenic tissues, and is considered suitable for further studies of tissue interactions in the tooth rudiment.

Endocrinology, 1976 Nov, 99(5), 1363 - 9
Secretion of a bone resorbing factor by chick thyroid glands in organ culture; Feinblatt JD et al.; The ability of the thyroid gland to secrete a bone resorbing factor in vitro was studied using glands obtained from 20-day-old chick embryos . The glands were incubated in a modified BGJ medium containing 1 mg/ml bovine serum albumin under 5% CO2-40% O2 at 37 C . The culture media were assayed in vitro by measuring the stimulation of the release of previously incorporated 45Ca from cultured 19-day fetal rat bone shafts over a 48 h period . The glands secreted a stimulator of bone resorption which did not appear to be parathyroid hormone (PTH) . The dose-response curve for the thyroid gland factor was not parallel to that obtained using PTH and secretion was not under calcium control . Neither thyroxine (T4) nor triiodothyronine (T3) produced a marked stimulation of bone resorption over a wide range of doses . Bone resorption stimulated by the thyroid gland factor was inhibited by calcitonin (CT) . Concentrations of TH and thyroid gland factor which were minimally effective when tested separately, produced a marked synergistic response when added together . This synergism was not seen when T4, T3, PGE1, or PGE2 were tested with PTH . Media obtained by culturing explants of embryonic chick liver, heart and muscle did not have bone resorbing activity . Secretion of the bone resorbing factor by thyroid glands was blocked by Indomethacin (10(-5)M) but the effects of the factor on bone were not blocked by this agent . These results suggest that the thyroid gland is capable of secreting a stimulator of bone resorption, possibly a prostaglandin, which is capable of synergizing with PTH, and which may represent a tissue factor which under certain circumstances may exert an influence on bone.

Infect Immun, 1976 Nov, 14(5), 1221 - 7
Isolation of a temperature-sensitive dengue-2 virus under conditions suitable for vaccine development; Eckels KH et al.; Dengue virus, type 2, in viremic human sera and after passage in cell cultures produces mixtures of small and large plaques when assayed in LLC-MK2 cells . Clones of dengue virus type 2 obtained by plaque selection in primary green monkey kidney cell cultures were tested for temperature sensitivity in vitro and for virulence by intracerebral inoculation of suckling mice . Sublines of a small-plaque clone were found to have lower nonpermissive temperatures than the parent virus by both plaque formation and release of infectious virus into the culture media . Small-plaque sublines were significantly less virulent in suckling mice than was the parent virus . Sublines from a large-plaque clone were not temperature sensitive and closely resembled parent virus mixed-plaque morphology . When small-plaque sublines were serially passaged using undiluted inocula, reversion occurred as evidenced by the appearance of large plaques and return of mouse virulence . Small-plaque virus could be maintained through several serial passages without reversion by using low-input inocula . Desirable passage history as well as temperature-sensitive and attentuation characteristics of the S-1 small-plaque subline make it appear suitable as a vaccine candidate virus.

J Physiol (Paris), 1976 Nov, 72(6), 815 - 32
Insulin secretion by human pancreas cultured for one year; Hollande E et al.; The pancreas of a child with intractable neonatal hypoglycemia was explanted in tissue culture on plasma clot and in monolayer culture after enzymatic dissociation . 1 . Cytological and immunoenzymatic studies of pancreas before explantation showed a very altered structure, suggesting a polyendocrine tumor composed mainly of B cells . Endocrine cells were present in the epithelium of duct-like structures . Large islets were often in continuity with these structures, suggesting islet budding from ducts, and supporting the hypothesis of the persistence of embryonic characteristics . 2 . In vitro, the pancreatic endocrine cells survived and proliferated; they were maintained for 362 days . During this time, they maintained their secretory capacity, as shown by radioimmunoassays of the culture media: the cells released insulin in rapidly decreasing amounts, then continued excreting low levels (5 to 60 muU/flask/day), in alternative periods of secretion and absence of secretion . 3 . Cytological study by light and electron microscopy of the cells in tissue and in monolayer cultures shows that they can undergo morphological changes, and may become epithelioid, fibroblastoid or round, while retaining their secreting activity . 4 . In long-term culture, the cells did not contain typical mature secretion granules . The hormone might be released into the medium by a clasmatosis mechanism . On the other hand, hormone excretion by vesicles originating from the rough endoplasmic reticulum is possible.

Biochim Biophys Acta, 1976 Oct 21, 450(1), 78 - 88
Effect of hyperlipemic serum on cholesterol accumulation in monkey aortic medial cells; Bates SR et al.; The effect of hyperlipemic monkey serum on cholesterol ester formation and accumulation in monkey aortic medial cells grown in tissue culture was studied . The cellular incorporation and esterification of free cholesterol was followed using the specific activity of serum labeled with free {14C}cholesterol while the cellular sterol content was analyzed by gas-liquid chromatographic techniqyes . The effects produced by hyperlipemic monkey serum (HMS) and normal monkey serum (NMS) were evaluated at both comparable percentage levels in the media and at equivalent exogenous cholesterol concentrations . When the two sera were adjusted to equal exogenous free cholesterol levels, the incorporation of free cholesterol by the aortic medial cells was related to the free cholesterol concentration of the culture media whether supplied by normal or hyperlipemic serum cholesterol . Under these conditions the total cholesterol content of the HMS-grown cells was 35% greater than that of NMS-grown cells, due to an elevation in free cholesterol of approximately 3 mug/mg cell protein and a 2- to 4-fold increase in esterified cholesterol . At similar percentage levels, the hyperlipemic serum stimulated a greater incorporation of free cholesterol into the monkey medial cells, accompanied by a 2-fold increase in the cellular esterification of this free cholesterol.

J Immunol, 1976 Oct, 117(4), 1331 - 5
Reversal of cell surface abnormalities of T lymphocytes in hodgkin's disease after in vitro incubation in fetal sera; Fuks Z et al.; The capacity of periphal blood lymphocytes from patients with untreated Hodgkin's disease to form E rosettes with sheep erythrocytes and to respond in vitro to PHA stimulation were found to be profoundly impaired . In 49% of the patients, the percentage of E rosette-forming cells (E-RFC) was more than two standard deviations below the mean for normal donors . Overnight incubation of the peripheral blood lymhocytes from these patients in culture media containing 20% fetal calf serum was followed by restoration of the percentage of E-RFC up to normal levels . Similar results have been observed after incubation in fetal human serum, but not in adult human AB serum or adult bovine serum . Incubation of peripheral blood lymphocytes from untreated patients in20% fetal calf serum also resulted in a remarkable restoration of their capacity to respond normally to PHA . Possible mechanisms involved in these reversible cell surface and in vitro lymphocyte function abnormalities in Hodgkin's disease are discussed.

J Embryol Exp Morphol, 1976 Oct, 36(2), 273 - 81
Biosynthesis of DNA, RNA and proteins by mouse embryos cultured in the presence of a teratogenic dose of chlorambucil; Sadler TW et al.; The effect of chlorambucil on the rates of DNA, RNA, and protein synthesis in mouse embryos was investigated using a system of whole embryo culture . Embryos were isolated on the 11th day of gestation (33 +/- 3 somites) and grown in culture media for periods of 4-8 h . Reichert's membrane and most of the placental tissue was removed leaving only the amnion and visceral yolk-sac surrounding the embryo . In the presence of teratogenic doses of chlorabucil (15 mug/ml) the rate of DNA synthesis was significantly decreased at 4 and 8 h . RNA and protein synthesis were not inhibited at either of these times . A trend toward decreasing rates of protein synthesis at some time beyond 8 h was noted, but not tested.

Can J Comp Med, 1976 Oct, 40(4), 408 - 13
In vitro effects of 2,4-dichlorophenoxy acetic acid (2,4-D) on bovine cells; Basrur SV et al.; Bovine fetal muscle cells were exposed to culture media containing 2 mg and 20 mg per liter of 2,4-dichlorophenoxy acetic acid (2,4-D) for varying intervals to determine the in vitro response of mammalian cells to this compound . The concentrations of 2,4-D used were comparable to those used in spray programmes although the residues normally found in pasture are much lower since 2,4-D is rapidly degraded under field conditions . Untreated and treated cultures were analyzed for total cell count, mitotic index and the percentages of differentiating and degenerating cells . The response of cultures to treatment was similar irrespective of the concentrations of 2,4-D used although in higher concentrations there was an initial drop in mitotic index . Other changes noted in treated cultures included an increase in differentiating and degenerating cells compared to those in control . The mitotic cells in treated cultures exhibited unipolar and tripolar spindles and a variety of other abnormalities including malorientation of the mitotic apparatus in relation to the axis of the cell . Myoblasts in initial stages of myogenesis were noted to be in mitosis in treated cultures suggesting that 2,4-D may have a stimulatory effect on myoblasts which in normal myogenesis are in post mitotic stage.

J Parasitol, 1976 Oct, 62(5), 676 - 9
Partial purification of amastigotes from cutaneous lesions of American leishmaniasis; Childs GE et al.; Amastigotes of Leishmania mexicana, L . mexicana amazonensis, L . brasiliensis, and L . enriettii were isolated from lesions in infected animals . Numbers of amastigotes recovered ranged from 1 X 10(7) to 7 X 10(8), depending on the strain of leishmania . Trypsinization disassociated the lesions and released the parasites . After 18 to 24 hr incubation at 37 C in tissue culture media with antibiotics, many of the intact host cells attached to the flask . Amastigotes were collected from the media in relatively pure preparations . Electron microscopy revealed no morphological alterations of the amastigotes and minimal contamination by membranes and cell fragments.

J Clin Pathol, 1976 Oct, 29(10), 934 - 7
Isolation of L-forms by blood culture; Brogan O; Culture media for the isolation of bacterial L-forms from the blood were studied . The most successful media had an osmolality of more than 1100 mosm/kg and this appeared to be a critical factor in determining success.

J Clin Invest, 1976 Oct, 58(4), 933 - 41
Rheumatoid factor-producing cells detected by direct hemolytic plaque assay; Vaughan JH et al.; Lymphocytes secreting anti-IgC antibodies, rheumatoid factors (RF), can be detected in the peripheral bloods, synovial fluids, and bone marrows of patients with seropositive rheumatoid arthritis by using a direct plaque-forming cell (PFC) assay with sheep erythrocytes sensitized with reduced and alkylated rabbit IgG hemolysin . The autospecific nature of the RF produced by RF-PFC was indicated by inhibition studies in which the order of patency was human IgG greater than rabbit IgG greater than bovine IgG . In metabolic studies puromycin, cycloheximide, and venblastine suppressed RF-PFC . Cyclic AMP and cyclic GMP were without effect . A need was recognized for using full tissue culture media during the cell separation and plaquing procedures to optimize detection of the RF-PFC . RF-PFC may appear in the blood of patients intermittently despite their continuing presence in the bone marrow . They have been found in the peripheral blood, especially during acutely exacerbating polyarticular synovitis, generalized vasculities, or generally active, aggressive disease . RF-PFC were found in synovial effusions of new or recrduescent acute synovitis . RF-PFC were observed to disappear from the peripheral circulation and the bone marrow during therapy with cytotoxic drugs . The data are consistent with the hypothesis that the appearance of RF-PFC in the peripheral blood represents an anamnestic response to transiently appearing antigen . The nature of the antigen is not specified . The bone marrow may be a site of origin of RF-PFC.

Proc Natl Acad Sci U S A, 1976 Oct, 73(10), 3680 - 4
Cellular cholesterol ester accumulation induced by free cholesterol-rich lipid dispersions; Arbogast LY et al.; The influence of lipoprotein composition on free cholesterol (RC) esterification and accumulation of esterified cholesterol (EC) was studied in cells of Fu5AH rat hepatoma exposed to culture media supplemented with (FC/P) ranging from 0.6 to 2.8 . In the presence of normal human serum, FC-lecithin dispersions witb a FC/P greater than one stimulated both the esterification of FC and the accumulation of cellular EC . Conversely, dispersions with FC/P values of approximately one had no effect on either FC esterification or the accumulation of EC when compared to cells grown in the absence of lipid dispersions . No stimulation of cellular response was obtained when FC-rich dispersions were added to cells in the absence of serum; however, stimulation was observed when cells were exposed to isolated human serum lipoproteins (very low density, low density, and high density) . With each lipoprotein, FC-lecithin dispersions with FC/P values less than one inhibited cholesteryl ester accumulation, FC-rich dispersions stimulated, and dispersions with FC/P values of approximately one had an effect equivalent to that of isolated lipproteins alone . These studies extablish that cellular FC esterification and EC content are not influenced solely by the FC concentration of medium, but rather respond to the ratio of FC to phospholipidl These studies also suggest that serum lipoproteins participate in the transfer of FC carried by FC-rich lecithin dispersions to cells.

In Vitro, 1976 Sep, 12(9), 635 - 8
Cytotoxicity of cysteine in culture media; Nishiuch Y et al.; When added to Eagle's Minimum Essential Medium supplemented with 10% bovine serum (MEM-10BS), 1mM cysteine was highly toxic to cultured cells . This toxicity was eliminated by (a) preincubation of the medium at 37 degrees C for 24 hr before use, or (b) presence of 5mM pyruvate . Similar results were obtained with freshly prepared CMRL 1066 supplemented with 10% bovine serum (CMRL-10BS), which contains 1.5 mM cysteine as an original ingredient . Medium L 15 supplemented with 10% bovine serum (L-10BS), which contains both 1 mM cysteine and 5 mM pyruvate, supported cell growth . On incubation of MEM-10BS supplemented with 1 mM cysteine (MEM-10BS-1CySH) or CMRL-10BS without cells for one day, the cysteine concentrations decreased to about one-tenth or less of the original concentrations . The cysteine concentration in L-10BS did not decrease so much on similar incubation . Pyruvate reduced the rate of disappearance of the cysteine in MEM-10BS-1CySH or CMRL-10BS as assayed with p-chloromercuribenzoate, although less than that in L-10BS . This effect of pyruvate was concentration dependent . These paradoxical effects of pyruvate on cysteine, i.e . the reduction of its cytotoxicity and the stabilization as an SH compound, are probably due to the formation of a dissociable complex between these two compounds, which is not cytotoxic and resistant to oxidation.

Can J Microbiol, 1976 Sep, 22(9), 1345 - 56
Degradation of {14C}photodieldrin by Trichoderma viride as affected by other insecticides; Tabet JC et al.; Various soil fungi were tested for their capacity to degrade the insecticide {14C}photodieldrin . Of nine species investigated, Trichoderma viride was the only one which degraded the insecticide to an appreciable extent into water-soluble, non-insecticidal compounds within 4-5 weeks . These products amounted to 32-41% of the radiocarbon applied to the culture media . The degradation was a function of live mycelia, which metabolized the insecticide and excreted water-soluble compounds into the culture media . Since soils usually contain a mixture of pesticide residues, the effects of several chlorinated hydrocarbon insecticides on the capacity of the fungus to degrade {14C}photodieldrin were studied . Thus, in fungal cultures treated with compounds structurally similar to photodieldrin, such as aldrin and dieldrin, only 4-17% of the applied radiocarbon was water-soluble and more photodieldrin remained . In controls, however, 35% of the applied radiocarbon was in the form of water-soluble products and less photodieldrin remained . The degradation of {14C}photodieldrin by T . viride, with time, was associated with a continuous decline of hexane-soluble radiocarbon and a steady increase of water-soluble metabolites, which appeared in the fungal media . The amount of hexane-soluble radiocarbon in mycelia was directly related to the fungal mass.

Cancer Res, 1976 Sep, 36(9 pt.1), 3269 - 73
Growth stimulation of cultured endothelial cells by tumor cell homogenates; Fenselau A et al.; Short-term cultures of endothelial cells from fetal bovine heart and aorta consistently display increased growth rates when crude tumor cell homogenates from the Walker 256 adenocarcinoma (ascites and solid forms as well as tumor cells from a suspension culture) are added to the culture media . The tumor-derived material produces growth stimulation in both sparse and confluient endothelial cell cultures . Homogenates of embryonic tissues and cultured cells show similar growth-promoting effects; corresponding material from various adult tissues is ineffective . The nonendothelial cells that were tested generally show no growth response or only a slight positive growth response to the tumor cell homogenates . The results indicate the feasibility of using this in vitro system as a paradigm of the tumor-induced vascularization process.

Fertil Steril, 1976 Sep, 27(9), 1067 - 77
Embryotoxicity of leukocyte extracts and its relationship to intrauterine contraception in humans; Parr EL et al.; Twelve human uteri containing intrauterine contraceptive devices (IUDs) and ten uteri without IUDs were obtained at hysterectomy . Samples of fluid were collected from the uterine lumina by absorbing the fluid in small pieces of lens paper . In the samples of luminal fluid we measured the concentration of beta-galactosidase, an enzyme which is present in human neutrophilic leukocytes and whose concentration in luminal fluid should correlate with the local inflammatory response to the intrauterine foreign body . In the samples of fluid from IUD-bearing uteri, the concentration of beta-galactosidase was significantly (P less than 0.0005) greater than that in luminal fluid from control uteri, the averages of the two groups differing by 3.8 units . To determine whether a foreign-body response of this magnitude could have any effect on preimplantation embryos, we cultured mouse embryos from day 4 to day 7 of development in culture media to which extracts of human leukocytes were added . All mouse embryos were killed when the culture media contained enough leukocyte extract to give beta-galactosidase concentrations of 0.5 unit or higher . Thus mouse embryos were killed by leukocyte extracts whose beta-galactosidase concentrations were actually less than the concentration of this marker enzyme measured in IUD uterine fluid . This comparison indicates that the luminal fluid in IUD-bearing uteri contains leukocyte break-down products in sufficient concentration to be lethal for preimplantation embryos.

Experientia, 1976 Aug 15, 32(8), 1045 - 7
Effects of prolactin and growth hormone on DNA synthesis of rat mammary carcinomas in vitro; de Iturri GC et al.; Explants derived from mammary carcinomas of DMBA-treated female Sprague-Dawley rats were cultured for 5 days in Medium 199 containing insulin and corticosterone . The addition of ovine prolactin to the culture media resulted in a consistent significant increase in H3-thymidine incorporation into DNA . DNA synthesis of explants treated with either ovine or human growth hormone was intermediary to prolactin-treated cultures and control cultures . A combination of prolactin and human growth hormone often increased DNA synthesis above either hormone alone, suggesting a possible growth synergism between these peptides.

Differentiation, 1976 Aug 3, 6(2), 105 - 11
Foetal rat pancreas in organ culture: effects of media supplementation with various steroid hormones on the acinar and islet components; McEvoy RC et al.; Foetal rat pancreas (20-day postcoitum) was grown in organ culture using a natural media (serum and chick embryo extract) . The media was supplemented with several adrenal and gonadal steroids; precursors, glucocorticoids, mineralocorticoids, estrogens, androgens and progesterone . The effects of the steroids on the pancreatic explants were quantitated biochemically by analysis of amylase and insulin in the incubated culture media and in the explanted pancreatic tissue, and morphologically by quantitative morphometric analysis . corticosterone, hydrocortisone, aldosterone and dexamethasone fully preserved the acinar cell component . The effect was concentration-dependent . Deoxycorticosterone and cortexolone had intermediate effects even at the highest concentration . Gonadal steroids had no effect on the acinar or islet component in this culture system . Some of the steroids inhibited the selective islet growth seen in the control explants as well as inhibiting insulin secretion . The relationship between these data and other work in this area is summarised . In addition, the possible implications of these data relating to normal in vivo pancreatic development are discussed.

J Natl Cancer Inst, 1976 Aug, 57(2), 409 - 14
Suppressor factor secreted by T-lymphocytes from tumor-bearing mice; Treves AJ et al.; Spleen cells from C57BL mice bearing the syngeneic carcinoma 3LL enhanced tumor growth . Tumor growth was also enhanced by a soluble factor found in the media of cultured spleen cells from tumor-bearing animals . This factor suppressed a protective immune response of the host and was found to be a product of T-lymphocytes . Removal of B-lymphocytes and macrophages did not prevent its appearance in the culture media, whereas removal of T-lymphocytes inhibited its appearance . Similar suppressor factors were obtained from C3H mice bearing the 3LL tumor and from mice with other tumors . The suppressing factor produced after the growth of 3LL tumor also enhanced the growth of other tumors . It could act on strains incompatible with the donor of the factor-producing cells . Hence tumor growth was possibly facilitated by soluble products of T-lymphocytes that were found in spleens of tumor-bearing mice and that nonspecifically suppressed immune defense mechanisms.

J Cell Sci, 1976 Aug, 21(3), 609 - 15
Amino acid metabolism of myeloma cells in culture; Roberts RS et al.; The growth of myeloma cells in Leibovitz medium supplemented with 20% serum was limited by the depletion of glutamine . A simple modification of the Leibovitz medium by increasing the concentrations of glutamine, lysine, isoleucine, leucine, sodium pyruvate, galactose, and vitamins resulted in over 100% increase in cell growth yield . The total myeloma protein produced by the cells was increased by approximately 90% in modified Leibovitz media . Analysis of spent culture media for 19 amino acids showed that the concentrations of 8 amino acids were reduced; those of 5 amino acids were increased and the other 6 did not change significantly.

J Immunol, 1976 Aug, 117(2), 548 - 54
Lymphokine-like factors produced by human lymphoid cell lines with B or T cell surface markers; Yoshida T et al.; In the present series of experiments we examined the ability of eight continuous cell lines derived from human lymphocytes to release lymphokines into their culture media . We have demonstrated that all could produce a macrophage migration inhibitory factor and a neutrophil chemotactic factor . This ability was independent of the B or T cell origin of the cell lines . As a further extension of previous reports, we report that the migration inhibitory effect does not seem to be due to cytotoxic effects as judged by trypan blue exclusion studies, and more important by the demonstration that the inhibition is reversible . The demonstration that neutrophil chemotactic activity is removable by treatment with a previously described and characterized anti-lymphokine antiserum suggests that this factor may be chemically similar to some of the lymphokines (migration inhibitory factor or chemotactic factor for macrophages) produced by antigen-activated guinea pig lymphocyte cultures . The failure to effect similar removal or inactivation of the macrophage migration inhibitory factor may simply reflect greater interspecies heterogeneity in this factor, or more interestingly, that this does not correspond to "classic" MIF . These results extend previous observations by defining further the capacity of lymphoid cell lines in continuous culture to produce mediators and by characterizing their relationship to conventional lymphokines produced by antigen or mitogen-activated lymphocytes.

J Clin Endocrinol Metab, 1976 Aug, 43(2), 374 - 86
Influence of purified plasma proteins on testosterone uptake and metabolism by normal and hyperplastic human prostate in "constant-flow organ culture"; Mercier-Bodard C et al.; Surgical samples of human prostate were explanted and submitted to constant-flow organ culture . The medium contained 3H-testosterone 50 nM, and except for controls, increasing concentrations of human serum albumin (HSA) or human sex-steroid-binding plasma protein (SBP) . At steady state, the explants were washed and homogenized, and the total radioactivity, radioactive testosterone, androstanolone (17 beta-hydroxyandrostan-3-one), androstane-3 alpha, 17 beta-diol, and androstane-3 beta, 17 beta-diol were determined after the addition of the corresponding internal 14C standards . From these data, testosterone uptake and metabolism were quantitated . The concentration of unbound testosterone in protein-supplemented culture media was measured separately by equilibrium dialysis . In control superfusions without protein, the tissue concentration of total radioactive steroids was equivalent to 182 +/- 18 (mean +/- SEM) pmoles/g of prostate . Androstanolone represented about 2/3, testosterone 1/10, and the two androstanediols together 1/10 of the total radioactivity . No difference was found between "normal" and hyperplastic prostate explants . In experiments with HSA (15-176 muM), is was observed that the uptake of radioactive testosterone in the prostate explants was decreased in direct proportion to the unbound testosterone fraction of the superfusion medium, but the proportions of testosterone metabolities in the superfused explants remained the same . In experiments with SBP (6-135 nM), the concentrations of unbound testosterone in the superfusion medium were reduced to the same levels as in the experiments with HSA . The reduction of tissue radioactivity was somewhat larger than that expected from the reduction of unbound testosterone in the superfusion medium for the concentrations of SBP less than 50 nM, and then remained approximately constant . In addition, SBP altered the metabolism of testosterone: the androstanolone/testosterone ratio in the prostate explants was critically dependent upon the SBP concentration in the superfusion medium . It is therefore suggested that, independent of its effect on the binding of testosterone, SBP has a direct effect on testosterone uptake and metabolism by the human prostate . The underlying mechanism is unknown.

Infect Immun, 1976 Aug, 14(2), 383 - 8
Factors influencing heat-labile Escherichia coli enterotoxin activity; Mundell DH et al.; In this study, conditions for production, detection, and storage of heat-labile Escherichia coli enterotoxin (LT) in culture filtrates from E . coli H-10407 were defined by using the adrenal tumor cell assay system . An enriched medium containing 0.6% yeast extract, 2% Casamino Acids, and 0.25% glucose buffered at pH 8.5 produced the highest LT activity of the various test media . In E . coli strain H-10407, LT activity was markedly decreased if the initial pH of the culture media was reduced to pH 7.5 or less . In contrast to E . coli P-263, if strain H-10407 was grown in the presence of mitomycin C there was no increase in LT production . Crude-culture filtrates containing LT can be stored at 4 degrees C for several days without an appreciable loss of activity; however, for long-term storage lyophilization or freezing at -70 degrees C is recommended.

Z Parasitenkd, 1976 Jul 27, 50(1), 57 - 66
{Immune mechanism of rats on Nippostrongylus brasiliensis in vitro (author's transl)}; Gerber HC et al.; The influence of humoral and secretory antibodies as well as cell supplements on Nippostrongylus brasiliensis was tested in vitro . Adult Sprague-Dawley-rats approximately 12 weeks of age were used in these experiments . For the in vitro tests the following culture media were used: 25% chicken-embryo-extract, sodium casein, pig liver extract and rat serum for the larval stages . The medium for the cultures of adult N . brasiliensis consisted of 5% yeast extract, 15% casein, 30% Krebs-Ringer-solution and 50% rat serum . Secretory antibodies were isolated from the rinsing fluid of the rat intestines by high pressure filtration (10 to 15 micron), then cleaning of the fluid through a Sephadex G 15 column and finally narrowing down through collodene capsules . Mast cells were isolated from the peritoneal cavity by Ficoll-gradient-centrifugation . Various test series were conducted with the addition of serum or secretory antibodies of repeatedly infected and immune rats to the medium . In these tests there was never a difference in the influence on growth nor a higher mortality rate of larval or adult N . brasiliensis in contrast to cultures where serum and secretory antibodies of non-infected animals were used . A 100%, degranulation of mast cells from infected rats occurred already within 14 to 22 hours in the cultures of adult N . brasiliensis . Variations were not noticed in the influence on the viability of N . brasiliensis kept in media for 10 days without or with cell supplements as well as sera of infected or not infected rats . The results from our experiments demonstrated that there was no variation in the influence on the development and a higher mortality rate of the larval stages and adult Nippostrongylus in media containing either sera and secretory antibodies of infected or not infected rats.

Proc Natl Acad Sci U S A, 1976 Jul, 73(7), 2429 - 33
Paradoxical effects of cycloheximide and cytochalasin B on hamster cell hexose uptake; Christopher CW et al.; Cellular regulation of hexose uptake was studied in cultures of NIL hamster cells . Enhancements of galactose uptake were elicited most strikingly by maintaining confluent NIL cultures in culture media devoid of glucose . These glucose-starved cultures showed up to 8- or 9-fold enhancements in the galactose uptake test . When these cultures were treated for extended periods with cycloheximide, the enhanced uptake was left unimpaired, whereas the uptake by glucose-fed cells, similarly treated with cycloheximide, was inhibited greater than 90% . Addition of glucose to these starved cultures resulted in a gradual decline of uptake rates to the unenhanced level (t1/2 approximately 3 hr) . In surprising contrast, when both glucose and cycloheximide were added simultaneously, the decline was arrested for at least 12 hr . If cytochalasin B (the specific inhibitor of hexose transport) was present, the uptake of galactose by both starved and fed cells was close to completely inhibited . By several criteria, cells maintained for 24 hr in medium containing both glucose and cytochalasin B were glucose-fed . Yet, when the cytochalasin B was removed, the cells were found to have enhanced rates of galactose uptake . The regulation of the hexose uptake system may therefore not be guided by the levels of glucose catabolites . Alternative mechanisms that may control hexose uptake are considered.

Prostaglandins, 1976 Jul, 12(1), 37 - 49
Characterization of prostaglandin production in tissue culture of rat renal medullary cells; Dunn MJ et al.; Renal medullary cells from the rat were used to establish a cell culture line . The morphologic characteristics of these cells were similar to renal medullary interstitial cells . They produced substantial amounts of PGE when provided with arachidonic acid or fetal calf serum . PGE production was inhibited 80-90% by indomethacin or meclofenamate . PGE release by the cells was sensitive to and stimulated by changing the culture media . Stable levels of PGE in the media could be achieved if media changes were avoided during the experimental period.

J Endocrinol, 1976 Jul, 70(1), 1 - 9
Incorporation of rat prolactin into rat milk in vivo and in vitro; Grosvenor CE et al.; Rat prolactin (11 i.u./mg) was continuously infused into the circulation of urethane-anaesthetized lactating rats for 35 min at doses of either 200 or 472 ng/min . The immunoreactive prolactin in both milk and plasma rose quickly during the first 20-25 min of infusion, then stabilized at similar levels over baseline (68 and 98 ng/ml for milk and plasma, respectively, with the 200 ng/min dose and 250 and 230 ng/ml, respectively, with the 472 ng/min dose) . The concentration of prolactin in plasma fell after the infusion was stopped, whereas that in the milk either did not fall at all or fell slightly to a new stabilized level . There was a rapid and extensive loss in the immunoreactivity of prolactin added to milk when rat milk was incubated in vitro (37 degrees C for 1-120 min) with 600 ng/ml of extracted pituitary prolactin (NIAMDD RP-1) or unit equivalent amounts of prolactin obtained from pituitary culture media (secreted prolactin, supplied by C.S . Nicoll) . Significantly greater amounts of added RP-1 prolactin were lost when it was incubated with milk obtained after 4 h than after 18 h of non-suckling . There was, however, no drop in endogenous immunoreactive milk prolactin levels (350-400 ng/ml) when rat milk was incubated with saline for 30 min . This suggests that milk prolactin obtained as a result of plasma transfer is different chemically from the milk prolactin resulting from the addition of either RP-1 or secreted prolactin to milk in vitro . Approximately 90% of 131I-labelled rat prolactin appeared in the trichloroacetic acid precipitable fraction after incubation (37 degrees C for 120 min) with milk obtained after 4 h of non-suckling in either the presence or absence of thiouracil (added to prevent binding of 131I or 131I-labelled fragments to milk protein) . The recovery was slightly less when 131I-labelled prolactin was incubated with milk obtained after 18 h of non-suckling . These data suggest that prolactin is quickly transferred from plasma into milk in direct relation to the plasma concentration . Once there, much of it appears to be retained by the milk perhaps chemically or physically bound; there is little, if any, degradation of the hormone . We conclude that the lactating the mammary gland may function normally as an excretory organ for prolactin.

Infect Immun, 1976 Jul, 14(1), 135 - 43
Thermal inactivation of rabies and other rhabdoviruses: stabilization by the chelating agent ethylenediaminetetraacetic acid at physiological temperatures; Michalski F et al.; Thermal inactivation of rabies and several other rhabdoviruses was studied using virus suspended in several different diluents . Rabies serogroup viruses were more stable than Kern Canyon or vesicular stomatitis viruses . Limited studies of two fish rhabdoviruses requiring low temperatures (less than 33 C) for replication indicated that they were not markedly more thermolabile than rabies virus . Bovine serum protein components in complex cell culture media stabilized virus at 56 C, but at temperatures of less than or equal to 37 C, sodium tris (hydroxymethyl)-aminomethane (NT) buffer containing ethylenediaminetetraacetic acid (EDTA) (NTE) was a much more efficient stabilizer of virus infectivity . Chelating agents EDTA and ethyleneglycol-bis-(beta-aminoethyl ether)tetraacetic acid were equally efficient in protection of rabies virus infectivity; the effect of each was lost when excess Ca2+ was added . Bovine serum in NT or NTE buffers produced a thermostabilizing effect at 37 C not provided by the same serum concentration in complex cell culture media . Bovine serum was more efficient than EDTA in stabilizing virus infectivity during repeated cycles of freezing and thawing.

Biochim Biophys Acta, 1976 Jun 23, 437(1), 94 - 105
N6,O2'-dibutyryl adenosine 3', 5'-monophosphate-stimulated release of proteoglycans from cultured immature rabbit ear cartilage; Shinmei M et al.; The stimulatory effects of N6, O2'-dibutyrl adenosine 3',5'-monphosphate on proteoglycan released from immature rabbit ear cartilage were studied in viltro . Cartilage incubayed in medium containing dibutyryl cyclic AMP resulted in a significant increase of proteoglycans released in concentrations above 0.5 mM . Theophylline (1 mM) which did not significantly stimulate proteoglycans released alone, was found to potentiate the action of this nucleotide . ATP, 5'-AMP and butyric acid in the presence of theophylline, did not stimulate proteolgycans released . The addition of protein or RNA synthesis inhibitors depressed proteoglycans released by dibutyryl cyclic AMP and theophylline . Gel chromatogrphic and chemical investigation of the proteoglycans released into the culture media in the presence of dibutylic cyclic AMP indicated a reduction in the proportion of protein associated with these complexes . This result, together with enzyme inhbitor studies, leads us to speculate that the observed action of dibutyryl cyclic AMP on rabbit ear cartilages may be mediated by the neutral proteases.

Proc Natl Acad Sci U S A, 1976 Jun, 73(6), 2023 - 7
Selenium is an essential trace nutrient for growth of WI-38 diploid human fibroblasts; McKeehan WL et al.; The trace element selenium is essential for clonal growth of diploid fibroblasts from human fetal lung (WI-38) in media containing small amounts of serum protein . Maximum growth stimulation is obtained when 30 nM neutralized selenious acid is added to a synthetic medium containing 1.5 mg/ml of dialyzed fetal bovine serum protein (equivalent to a 3% serum concentration) . Serum appears to be a source of selenium in most culture media, since higher concentrations of serum protein or whole serum mask the selenium requirement of WI-38 cells . Selenium is also required by a Chinese hamster cell line that can be grown in a protein-free synthetic culture medium.

In Vitro, 1976 Jun, 12(6), 436 - 41
Occurrence of minute ring-shaped nucleoprotein particles in culture media conditioned by mammalian cells; Ashkenazi-Ezra R et al.; Conditioned media of 21 mammalian cell lines have been examined by electron microscopic and biochemical techniques for the presence of a minute ring-shaped nucleoprotein particle (RSP) . Electron microscopy showed the presence of RSP in 20 of the 21 cell lines tested . Comparative analyses of sex cell cultures for radioactive RSP, following 3H-thymidine incorporation, suggested that quantitative differences exist in the amount of RSP detectable in conditioned media of normal and pathologic cells . The cell lines tested included cells of different morphologies, origin and function

Am J Anat, 1976 Jun, 146(2), 133 - 49
Fetal and neonatal rat pancreas in organ culture: age-related effects of corticosterone on the acinar cell component; McEvoy RC et al.; Fetal rat pancreas (ages 16 to 22 days postcoitum) and neonatal pancreas (4 days postnatal) were grown in organ culture for four days . The medium consisted of chick embryo extract and rooster serum either with or without the addition of corticosterone (3 X 10(-5) M) . Acinar cell differentiation was assessed using quantitative light microscopic linear scanning of tissue sections and enzymatic analysis of amylase in the culture media and in the explants . In the younger fetal tissue of 16 and 18 days postcoitum exocrine differentiation continued in vitro . The effect of corticosterone was an enhancement of the degree of in vitro differentiation . Even with corticosterone, however, the degree of differentiation in vitro was less than that observed during a similar period in vivo . In differentiated pancreas (20- and 22-day fetal neonatal) the acinar pancreas degenerated under control conditions and a selective growth of the endocrine pancreas was observed . The addition of corticosterone to the media resulted in a maintenance of the differentiated state of the acini except in 22-day fetal tissue in which the acini were not preserved . The differences between these results and the work of other investigators and the possible in vivo role of adrenocorticosteroids in exocrine pancreatic differentiation is discussed.

J Med Genet, 1976 Jun, 13(3), 223 - 8
Supravalvular aortic stenosis-infantile hypercalcaemia syndrome: in vitro hypersensitivity to vitamin D2 and calcium; Becroft DM et al.; The incidence of cytoplasmic metachromasia has been studied in cultures of skin fibroblasts derived from 6 cases of the syndrome of supravalvular aortic stenosis, characteristic facies, and mental retardation which in many instances represents the late normocalcaemic stage of the severe form of infantile hypercalcaemia . The percentage of metachromatic cells (mean positivity 7.3%) was significantly higher than in control cultures . The addition of vitamin D2 and calcium to culture media caused a highly significant increase in metachromatic cells (mean positivity in supplemented media 16.1%) compared with a lesser increase in controls . These findings strengthen previous suggestions that there is a genetically determined hypersensitivity to vitamin D in some cases of the syndrome . A multifactorial aetiology is proposed, dependent on a variable genetic susceptibility of fetal connective tissues to a non-physiological effect of D vitamins and a variable level of maternal vitamin D nutrition.

Ann Immunol (Paris), 1976 Jun-Jul, 127(3-4), 519 - 30
Evidence for multipotentiality of antibody synthesizing cells; Liacopoulos P et al.; Immunization of mice with two unrelated antigens or antigenic determinants regularly results in the appearance of hemolytic plaque forming cells (PFC) of each specificity and of some PFC (1-4%) reacting to both determinants . Micromanipulation of individual double PFC appearing after immunization with trinitrophenyl conjugated sheep erythrocytes (TNP-SRBC) into media containing indicator erythrocytes (native SRBC and TNP-horse RBC) and a soluble specific inhibitor (TNP-BSA or soluble SRBC antigen), showed that the specific inhibitor suppressed the lysis of the corresponding indicator but did not interfered with the lysis of the unrelated indicator . Persistance of one specific activity in spite of complete inhibition of the other indicated that these double PFC synthesize two different antibody molecules . The destiny of double cells was studied in individual cell cultures by micromanipulating them into wells containing heavily irradiated normal mouse spleen cells and both determinants (TNP-SRBC) . Those double cells which divided they generated in 24 hours monospecific daughter PFC (either anti-TNP or anti-native SRBC) . Individually cultured monospecific PFC from mice immunized with one antigen (native SRBC) generated daughter PFC of the same specificity . By contrast, monospecific PFC from mice immunized with TNP-SRBC generated PFC of either the same specificity of (more frequently) of the one the other specificity . Thus, immunization with substituted erythrocytes resulted in the development of three clonotypes: two clones each composed only of cells of either specificity (amphispecific clonotype) . For the maintenance of this amphispecific clone, continuous presence in the culture media of both antigenic determinants was necessary . Removal of the one for 48 hours (but not for 24 hours) abolished the generation of daughter cells of the same specificity.

Med Klin, 1976 May 14, 71(20), 865 - 8
{Mycoplasms in tumors of patients with carcinomata of the collum and its preliminary stages (author's transl)}; Rutke J et al.; Within one year smears were taken from 190 patients who had not been pretreated in our oncological department . The culture media established were examined with regard to Mycoplasms . A Mycoplasma population was stated in 27 women . In the cases of carcinomata of the collum and in its preliminary stages, Mycoplasma infestation was about twice as frequent as in all other cases of genital carcinomata . Discussion is made if Mycoplasms are possible or promoting factors during the development of carcinomata of the collum.

Biochim Biophys Acta, 1976 May 3, 432(2), 223 - 36
Control of the formation of extracellular ribonuclease in Neurospora crassa; Hasunuma K et al.; A finding was made that a species of ribonuclease is released into mycelial culture media when a wild-type strain of Neurospora crassa was grown on limiting amounts of phosphate . The ribonuclease activity in the fully derepressed state extends to about 60 to 100 fold of that in the repressed state . The synthesis of the ribonuclease was inhibited by the addition of rifampicin, cycloheximide or orthophosphate . Three molecular species of the ribonuclease were found . Two enzyme fractions showing larger molecular weights were suspected to be aggregates containing the enzyme showing the smallest molecular weight (molecular weight of 10 300) . All three fractions showed pH optima of around 7, preferential hydrolysis of polyguanylic acid and poor hydrolysis of guanosine 2',3',-cyclic monophosphate . These characteristics were the same as those of ribonuclease N1, and it was suggested that ribonuclease N1 is a repressible extracellular enzyme . Mutations in the genes nuc-1 and nuc-2 caused loss of ability to derepress this enzyme, but heterokaryon between them partially restored the ability . The nuc-1 mutation was epistatic to the nuc-2 alleles which are partly constitutive in the ribonuclease production.

J Immunol, 1976 May, 116(5), 1324 - 31
Naturally soluble tumor antigens from guinea pig hepatomas: isolation and partial characterization; Detrick-Hooks B et al.; Naturally soluble tumor antigens were detected in the ascites fluid of guinea pigs bearing an ascites tumor and from exhausted tissue culture media of cultured tumor cells . Two antigenically distinct cell lines of diethylnitrosamine-induced strain-2 guinea pig hepatomas (line-10 and line-1) served as the source of tumor antigens . Tumor antigen activity was detected by four different techniques: immunodiffusion, inhibition of complement-mediated cytotoxicity, inhibition of membrane immunofluorescence, and delayed cutaneous hypersensitivity . With syngeneic tumor-specific antiserum, line-10 guinea pig tumor antigens were detected by immunofluorescence in the concentrated ascites and tissue culture fluids . With a xenogenic antiserum, demonstrated to be tumor specific, line-10 tumor antigens were detected not only in the concentrated ascites and tissue culture fluids but also in two of the partially purified fractions of these fluids . When the line-10 concentrated ascites and its fraction I were subjected to ultracentrifugation at 300,000 x G for 1 hr, the antigen activity was retained in the supernatant and thus by this criterion the tumor antigens detected in these samples are soluble . Immunodiffusion data indicate that more than one antigen is present in the line-10 system since three lines of precipitation were detected when line-10 concentrated ascites was reacted with the line-10 tumor-specific antiserum . In contrast to this, the line-10-concentrated tissue culture fluid displayed only one line of precipitation . Although tumor antigens could not be demonstrated in the other antigenically distinct tumor cell line, line-1, by immunodiffusion or inhibition of membrane immunofluorescence, inhibition of complement-mediated cytotoxicity was able to detect tumor antigens in the line-1 concentrated ascites and tissue culture fluids.

In Vitro, 1976 May, 12(5), 363 - 72
Studies on normal human bone marrow cultures in controlled environment thin-film culture systems; Gailani S et al.; Two thin film culture systems, the controlled environment steady state system (SS) and the rocker tube configuration of that system (RT), were used to identify some of the conditions that appear to maintain morphologic and functional characteristics of cells of human bone marrow explants in vitro . The systems configuration assured continual gassing, control and easy monitoring of the cultures . Cytocentrifuge preparations of media of specimens cultured in RT disclosed, though in decreasing numbers, various hematopoietic cells for periods exceeding one month . Hematopoietic cells shed from specimens cultured in the SS system were retained in the culture tubes; cells of the myelocytic series predominated for the first two weeks while an increasing number of monocytes and macrophages appeared in the media of of older cultures . Histologic examination of cultured explants disclosed preservation of the marrow architecture and the persistence of hematopoietic cells . Specimens cultured in RT tubes tended to be less cellular than similar cultures placed in dialysis bags or as cultured in the SS system . Immunoglobulins (Ig) were released into the culture media at a constant rate throughout the period of culture . Specimens that were cultured at a controlled pH of 7.4 released 2 to more than 4 times as much Ig as similar specimens maintained at a pH level of 7.1 . There were no definitive differences in Ig levels in the cultures maintained at comparable pH levels and overlaid with various CO2 concentrations, i.e . 2%, 5%, 10% similarly, no differences in Ig levels were found in specimens cultured in media containing fetal bovine sera as opposed to horse sera.

Hum Genet, 1976 Apr 15, 32(1), 49 - 64
Comparative studies on the specificity of anticlastogenic action in human lymphocytes in culture; Gebhart E et al.; Comparative studies on human lymphocyte cultures yielded a certain specificity of the anticlastogenic action of the SH compounds l-cysteine, cysteamine, and beta-aminoethylisothiouronium (AET) as well as of the amide l-asparagine and the amino acid l-methionine . This specific anticlastogenic activity manifested itself in specific changes of the spectrum of aberration types induced by the clastogens and of the pattern of intercellular distribution of the induced aberrations . It was clearly dependent on the concentration of the anticlastogens but was also influenced by the used clastogen . The use of different culture media yielded some quantitative influences on the anticlastogenic activity, but fundamental changes in the spectrum of anticlastogenic action have not been observed except with cysteamine . The patterns of activity ascertained for the different anticlastogens specifically differed from those changes in the spectrum and pattern of distribution of aberrations induced by a mere reduction of the concentration for instance of Trenimon . Therefore a direct reaction between the protectors and the clastogen Trenimon as the cause of the observed anticlastogenic action was again excluded . The presented data are also discussed under the aspects of the hypotheses of aberration induction as well as of their importance for further antimutagen research.

Cell Tissue Res, 1976 Apr 9, 167(4), 493 - 514
The effects of testosterone, 5alpha-dihydrotestosterone, 3alpha-androstanediol, and 3beta-androstanediol on epithelial fine structure of the rabbit epididymis in organ culture; Hoffman LH et al.; The fine structure of the corpus epididymidis of the rabbit has been studied following organ culture . Various modifications of tissue preparation and culture conditions were examined to obtain good maintenance of cellular integrity as well as to preserve sperm fertilizing ability . After 5 to 7 days in culture in the absence of hormonal support, the epididymal epithelium showed signs indicative of cellular regression . Such changes included shrinkage of the cells, loss of the border of stereocilia, decrease in smooth endoplasmic reticulum, and an increase in autophagic vacuoles . The presence of androgens in culture media prevented cellular regression to varying degrees, depending on the hormone utilized . With regard to maintenance of cellular integrity, potency of the androgens tested was as follows: 5alpha-dihydrotestosterone greater than or equal to 3alpha-androstanediol greater than testosterone greater than 3beta-androstanediol . Addition of insulin to dihydrotestosterone-containing cultures resulted in no improvement in maintenance . Phagocytosis of spermatozoa by epithelial cells was observed in cultured tubules and the degree of spermiophagy was inversely proportional to successful maintenance of fine structural characteristics of epithelial cells . The morphological findings reported here correlate well with the fertilizing ability of spermatozoa from cultured epididymis as reported in an accompanying communication.

Mutat Res, 1976 Apr, 40(2), 131 - 8
Cytogenetic study of DDT on human lymphocytes in vitro; Lessa JM et al.; The cytogenetic effect of DDT on human blood cultures, in vitro, was investigated . Two types of experiment were carried out: one, in which the DDT concentrations found in the culture media were similar to those found in the plasma of individuals of the Brazilian population (0.06--0.20 mug/ml); in the second experiments, doses ranging from 1 to 15 mug/ml were used . No correlation was found between DDT doses and cells with chromosomal aberrations . The Poisson test of comparison between means showed that at certain DDT concentrations (0.20, 4.05 and 8.72 mug/ml) the proportion of cells with structural aberrations was significantly greater than in the controls.

Cancer Res, 1976 Apr, 36(4), 1299 - 304
Controlled environment culture of bone marrow explants from human myeloma; Gailani S et al.; Bone marrow biopsy specimens from patients with myeloma were cultured in either 1 of 2 thin-film culture systems, a controlled environment steady state system or a rocker tube configuration of the system, for periods up to 42 days . Both functional and morphological characteristics of the myeloma cells were well-maintained in these systems . Cytocentrifuge preparations of the culture media disclosed hematopoietic cells that included from 5% to almost 100% plasma cells . Histological examination of the cultured specimens disclosed infiltration of the marrow with myeloma cells . Myeloma proteins were released at a steady rate throughout the period of culture after the 1st 4 days . Bone-resorbing activity was demonstrated in the culture media in 7 of 9 myeloma culture media and was well maintained, particularly during the 1st week of culture . This activity was associated with severe osteolytic lesions in the donor patient and marked infiltration of the cultured specimen by myeloma cells . The potential use of these organ culture systems for the further definitive identification of the factor responsible for bone destruction in myeloma is discussed.

Sabouraudia, 1976 Mar, 14(1), 1 - 4
The isolation of Botryodiplodia theobromae from a nail lesion; Restrepo A et al.; Botryodiplodia theobromae not known to produce onychomycosis was repeatedly recovered from a healthy woman with evident lesions in a toe nail . Mycelial fragments were observed in the scales and the fungus was isolated in cycloheximide-free culture media . The report indicates that many fungi, hitherto considered non-pathogens, may still be able to colonize a vaiety of human tissues.

Arch Neurol, 1976 Mar, 33(3), 180 - 2
Desmosterol in human and experimental brain tumors in tissue culture; Weiss JF et al.; Desmosterol, a possible chemical indicator of brain tumors, was detected in cells of neurogenic, nitrosourea-induced rat tumors (neurinomas and gliomas, C6 cell line) and in human astrocytomas grown in lipid-poor media . A further increase in the amount of cell desmosterol was obtained when triparanol was added to media containing delipidized serum . Cholesterol was replaced almost completely by desmosterol in tumor cells grown in media containing nontoxic levels of 20,25-diazacholesterol . Desmosterol did not accumulate when these inhibitors of desmosterol-reductase were added to culture media containing cholesterol and other lipids (whole fetal calf serum) . The results demonstrate that tumors of the nervous system grown in tissue culture are capable of sterol synthesis, and indicate that a central mechanism of cholesterol synthesis is operative in these cells, which may be related to the availability of exogenous cholesterol . It is concluded that these findings are relevant to clinical studies on the use of cholesterol inhibitors as tools for the detection of brain tumor activity.

Somatic Cell Genet, 1976 Mar, 2(2), 141 - 53
beta2-microglobulin locus on human chromosome 15; Faber HE et al.; We have developed an autoradiographic/electrophoretic assay capable of distinguishing mouse and human beta2-microglobulin (beta2m) in spent culture media . The method is applicable to mouse and human lines and to hybrid cell lines made from them . With this technique, mouse/human hybrid cell lines were tested for the presence of human beta2m . Isozyme and karyotype analyses were also carried out with the hybrids . The combined results of these studies show that the structural gene for human beta2m is on the long arm of chromosome 15.

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