Effects of acronycine on nucleic acid synthesis and population growth in mammalian tumor cell cultures

Acronycine — an alkaloid with antineoplastic activity against a wide range of experimental tumors — at concentrations of 0.5-12 μg/ml rapidly inhibits RNA synthesis in L5178Y mouse lymphoma and IRC rat monocytic leukemia cultures. Culture growth is arrested only at acronycine concentrations which markedly inhibit RNA synthesis. DNA synthesis is inhibited at rather higher concentrations but this is not a prerequisite of the arrest of growth. It is suggested that the arrest of growth may be a consequence of the inhibition of RNA synthesis. In both cultures arrest of growth coincides with the appearance of many cells with two apparently normal nuclei. Cells are not arrested in mitosis. It is shown these binucleated cells very probably arise from an inhibition of cell cleavage. Studies with synchronized cultures show that at low drug concentrations, more than one cell cycle may elapse before growth is arrested and binucleated cells appear, indicating the effect on cytokinesis is not immediate. The results suggest that the arrest of growth may be a result of a slow depletion of a component essential for cell cleavage. The disturbance at division is a major factor in arresting growth at low drug concentrations. At higher acronycine concentrations, when RNA synthesis may be inhibited by 80–90%, the cytotoxic effects appear earlier and are less specifically directed at cytokinesis; DNA synthesis is then also rapidly and markedly inhibited.     

人参(Panax ginseng)根系分泌物成分对人参致病菌的化感效应

采用室内培养结合生物学测定的试验方法,研究了不同浓度人参(Panax ginseng)根系分泌物成分苯甲酸、邻苯二甲酸二异丁酯、十六酸和2,2-二(4-羟苯基)丙烷对人参立枯丝核菌(Rhizoctonia solani)、黑斑菌(Alternaria panax)、疫病菌(Phytophthora cactorum)、菌核菌(Sclerotinia schinseng)、锈腐菌(Cylindrocarpon destructans)和绿色木霉菌(Trichoderma viride)菌落生长及孢子萌发的化感效应.结果显示,不同浓度人参根系分泌物成分对人参致病菌及绿色木霉菌的化感效应存在显著差异.苯甲酸浓度与人参立枯丝核菌、菌核菌和锈腐菌菌落生长以及人参黑斑菌、锈腐菌孢子萌发呈负相关,与人参黑斑菌、绿色木霉菌菌落生长呈正相关;对人参疫病菌菌落生长的化感效应表现为低浓度和高浓度抑制,中浓度促进.邻苯二甲酸二异丁酯浓度与人参立枯丝核菌、黑斑菌、菌核菌和绿色木霉菌菌落生长以及人参黑斑菌孢子萌发呈负相关;对人参锈腐菌菌落生长和孢子萌发表现为低浓度和高浓度抑制,中浓度促进;对人参疫病菌菌落生长表现为低浓度和中浓度抑制,高浓度促进.2,2-二(4-羟苯基)丙烷浓度与人参立枯丝核菌、黑斑菌、疫病菌、绿色木霉菌菌落生长以及人参黑斑菌、锈腐菌孢子萌发呈负相关;对人参菌核菌、锈腐菌菌落生长表现中浓度促进,高浓度抑制.十六酸浓度与人参锈腐菌、疫病菌和绿色木霉菌菌落生长呈正相关,与人参锈腐菌孢子萌发呈负相关,对黑斑菌孢子萌发表现为中浓度抑制.4种根系分泌物的等量混合物浓度与人参致病菌及拮抗木霉菌菌落生长速率呈负相关.     

Russian Article pp. 27–32

The influence of various concentrations of lysine and threonine on the growth rate (μ), homoserine- deficient and prototroph revertant at their joint growth in liquid fermentation medium has been investigated. It has been established that even relatively small concentrations of threonine (25 mg/l) at the background of lysine concentration exceeding 10 g/l reduce μ of prototrophs, but do not influence μ of auxotrophs. It is supposed that the addition of threonine at the background of comparatively large concentrations of lysine is one of the factors creating selective conditions under which an auxotroph producer has advantage over prototroph revertant.     

Qualitative behavior of a variable-yield simple food chain with an inhibiting nutrient

A simple food chain which consists of nutrient, prey and predator in which nutrient is growth limiting at low concentrations but growth inhibiting at high concentrations is investigated in this study. It is assumed that the nutrient concentration is separated into internal and external nutrient concentration and only the internal nutrient level is capable of catalyzing cell growth. It is shown that the dynamics of the system depend on thresholds R(0) and R(1). With inhibition, there exist initial conditions for which the predator becomes extinct but not the prey when R(0)<1. If R(0),R(1)1, the system is uniformly persistent even in the inhibited environment.     

The role of erbB-2 and its ligands in growth control of malignant breast epithelium.

The erbB-2 (HER-2, neu) protooncogene is overexpressed on the surface of about 25% of human breast cancers. It is homologous to epidermal growth factor receptor and a putative growth factor receptor. Overexpression in breast, ovarian and gastric cancers is associated with a worse prognosis. We have recently discovered two ligands for this receptor. They both induce receptor phosphorylation. At low concentrations both induce clonogenic growth of overexpressing cells; at higher concentrations both are growth inhibitory. Both can overcome the inhibitory effects of both monoclonal antibodies directed against the ligand binding site and soluble extracellular domain. These ligands may form an attractive basis for antitumor therapy.     

Season variability of iron effects on periodic culture of microalgae Dunaliella viridis Teod.

Effects of different iron concentrations (final concentrations of iron in Artari's medium: 3.7, 37.0, 74.0, and 185.0mmol·L-1) on growth rate and contents of protein, triacylglycerides, and β-carotene in Dunaliella viridis cells at cultivation in different months were investigated. It was shown that the dose-dependent effects of iron were notable for season variability. In the 1stexperimental series (October, 2007), iron at researched concentrations did not affect growth rate of culture and protein, triacylglyceride, and [3-carotene contents in cells. In experimental series conducted respectively in Novem-ber 2007, December 2007, and February 2008, the dose-dependent stimulation of microalgae growth was observed. For each of these experimental series, there were particular dose dependences of protein, triacylglyceride, and β-carotene contents in microalgae cells at cultivation on media with iron at different concentrations. Meanwhile, for all of the four experimental series conducted in different months, variability of growth rate and analyzed parameters of microalgae Dunaliella viridis as control (cultivation without iron) was shown. It is suggested that these functional differences of control cultures of microalgae in different months caused variability in the dose-dependent effects of iron in a Dunaliella viridis culture. The possibility of iron usage for increasing microalgae biomass and for enriching it by β-carotene in Dunaliella viridis culture with initial low productivity and low β-carotene content is considered.     

Effect of argillaceous minerals on the growth of phosphate-mobilizing bacteria Bacillus subtilis

It was shown that the argillaceous minerals montmorillonite and palygorskite at concentrations within 0.2-1.0% considerably accelerate the growth of phosphate-mobilizing bacteria Bacillus subtilis grown in media with hardly soluble Ca3(PO4)2 as the sole source of phosphorus. The most notable effect of these minerals was recorded at concentrations within 0.5-1.0%. The effect of argillaceous minerals in the colloidal form on bacterial growth was more pronounced than that of the powdered ones. An increase in montmorillonite or palygorskite concentrations to 2% is accompanied by the inhibition of the growth of the phosphate-mobilizing strain. At such concentrations the minerals adsorb ca. 22% of the glucose and 11.3% of the phosphate added to the nutrition medium.     

The inhibitory effects of pyrocarbonic acid diethyl ester on lactobacillus casei and its phage Jl – A novel strategy for phage control in technical fermentation processes

The effects of pyrccarbonic acid diethyl ester (PADE) on Lactobacillus casei Sl and its phage Jl was investigated in relation to the control of phages in the dairy industry and other technica fermentation processes. PADE exhibited a bacteriostatic effect at 0.5 to 8 mM and a bactericidal effect at 10 mM or higher. It inhibited the growth of the phage at its bacteriostatic and bactericidal concentrations. The growth inhibition of the phage was reversible at the bacteriostatic concentrations but complete and irreversible at the bactericidal concentrations. PADE inactivated the free phage within several minutes; 10 and 30 mM of PADE inactivated 90 and 100%, respectively, of the phage. It completely decomposed into ineffective components in several minutes. The bacteria grew almost normally when they were inoculated after the complete decomposition of PADE. These four characteristics of PADE – its bactericidal effect, its inhibitory effect on phage growth, its phage-inactivating effect and its decomposition – suggest a novel strategy for phage control in technical fermentation processes, including the dairy industry.     

Phosphate-limited continuous culture of Rhodotorula rubra: kinetics of transport, leakage, and growth.

The phosphate-limited growth kinetics of Rhodotorula rubra, a small yeast of marine origin, were examined by analysis of 32P distributions in continuous cultures. Isotope relaxation procedures were used to identify unidirectional flows of Pi and organic phosphate among compartments modeled during growth. The concentrations of phosphates in these compartments at various growth rates were used, together with attendant flows, to produce a mathematical model of growth. Both Pi and phosphate-containing metabolic intermediates leaked from cells during growth. Total leakage ranged from 4 to 10% of influx and was comprised mostly of Pi. Transport capacity was at least 10 times that required for growth at saturating Pi concentrations, so that influx was linear with concentration during growth. This led to the realization that the curvature of Monod plots (Kmu = 12 nM mumax = 0.18/h, and the threshold At = 2.5 nM) is due to change in yield with growth rate. Growth rate related to Pi by the affinity, aA (= 0.43 liter/mg of cells.h) of cells for Pi and the growth rate-dependent yield. It was also specified by a series of kinetic constants that specified flow among the various compartments and equilibrium compartment concentrations as they were set by extracellular Pi. The importance of leakage by healthy cells to the organic chemistry of aquatic systems is noted.     

Growth of mixed cultures on mixtures of substitutable substrates: the operating diagram for a structured model

The growth of mixed microbial cultures on mixtures of substrates is a problem of fundamental biological interest. In the last two decades, several unstructured models of mixed-substrate growth have been studied. It is well known, however, that the growth patterns in mixed-substrate environments are dictated by the enzymes that catalyse the transport of substrates into the cell. We have shown previously that a model taking due account of transport enzymes captures and explains all the observed patterns of growth of a single species on two substitutable substrates (J. Theor. Biol. 190 (1998) 241). Here, we extend the model to study the steady states of growth of two species on two substitutable substrates. The model is analysed to determine the conditions for existence and stability of the various steady states. Simulations are performed to determine the flow rates and feed concentrations at which both species coexist. We show that if the interaction between the two species is purely competitive, then at any given flow rate, coexistence is possible only if the ratio of the two feed concentrations lies within a certain interval; excessive supply of either one of the two substrates leads to annihilation of one of the species. This result simplifies the construction of the operating diagram for purely competing species. This is because the two-dimensional surface that bounds the flow rates and feed concentrations at which both species coexist has a particularly simple geometry: It is completely determined by only two coordinates, the flow rate and the ratio of the two feed concentrations. We also study commensalistic interactions between the two species by assuming that one of the species excretes a product that can support the growth of the other species. We show that such interactions enhance the coexistence region.     

A Switch Role of Src in the Biphasic EGF Signaling of ER-Negative Breast Cancer Cells

It is well established that epidermal growth factor (EGF) is a potent mitogen in cells expressing EGF receptor (EGFR). However, a body of evidence indicated that the effects of mitogenic EGF signaling exhibit a non-monotonic, or biphasic dose response curve; EGF at low concentrations elicits a mitogenic signaling pathway to stimulate cell proliferation while at high concentrations, EGF inhibits cell growth. However, the molecular mechanism underlying this paradoxical effect of EGF on cell proliferation remains largely unknown. Here, we investigated the molecular mechanisms underlying the biphasic EGF signaling in ER-negative breast cancer MDA-MB-231 and MDA-MB-436 cells, both of which express endogenous EGFR. We found that EGF at low concentrations induced the phosphorylation of the Src-Y416 residue, an event to activate Src, while at high concentrations allowed Src-Y527 phosphorylation that inactivates Src. EGF at 10 ng/ml also induced phosphorylation of the MAPK/ERK and activated cyclin D1 promoter activity through the Src/EGFR/STAT5 pathways but not at a higher concentration (500 ng/ml). Our results thus demonstrated that Src functions as a switch of EGF signaling depending on concentrations of EGF.     

Effect of cycloheximide on IAA-induced proton excretion and IAA-induced growth in abraded Avena coleoptiles

IAA-induced proton excretion in peeled or abraded oat ( Avena saliva L. cv. Victory) coleoptiles is closely associated with IAA-induced growth. It was attempted to separate these two processes by using cycloheximide to inhibit them differentially. Growth of abraded coleoptile segments was measured by a shadow graphic method, and their IAA-induced acidification of the external solution was monitored with a pH meter. IAA stimulated proton excretion in abraded Avena coleoptile segments after a 13 min lag. IAA-induced proton excretion was inhibited within 5 min by cycloheximide at concentrations of 1.8 × 10⁻⁶, 3.6 × 10 or 3.6 × 10⁻⁵ M. Cycloheximide at these concentrations, added within 4 min of IAA, prevented IAA-induced acidification of the medium for at least 60 min. However, it did not prevent IAA-induced growth during this time. It is concluded that some of the initial IAA-induced growth seen in Avena coleoptiles is independent of detectable IAA-induced proton excretion.     

Assessment of nutrient availability and limitation using macroalgae

Discs of the macroalga,Ulva lactuca L., were transplanted around an ocean outfall and at a reference site in Køge Bay, Denmark, to assess the influence of the outfall on the nutrient availability. At 2-wk intervals, samples were collected and analyzed for growth, nitrogen, and phosphorus content.The tissue concentrations of nitrogen and phosphorus decreased with distance to the outfall, showing that the tissue concentrations are suitable for monitoring nutrient availability in coastal areas and provide a time-integrated measure of the nutrient availability. The lowest tissue concentrations of nitrogen were recorded at the reference station, where the internal concentrations generally were below the critical concentration level, showing that nitrogen limited the growth. At the station located close to the outfall, the flux of nitrogen was sufficient to maintain the maximum growth rate. The tissue concentrations of phosphorus were only below the critical concentration level on one occasion, and the result showed a net uptake throughout the study period.It was concluded that in the Køge Bay, nitrogen was the main limiting factor for macroalgae growth during the summer. The applicability of tissue concentrations for assessment of nutrient availability is discussed and it is considered that the method, when evaluated against established critical concentrations, provides a valuable tool for assessing ecosystem health with regard to eutrophication.     

Purification of a fibroblast growth factor from bovine pituitary.

The purification from bovine pituitary gland of a growth factor responsible for the control of animal cell division in tissue culture is reported. This growth factor is a polypeptide of 13,300 molecular weight and is homogeneous when analyzed by polyacrylamide gel electrophoresis, carboxymethyl-Sephadex gradient elution chromatography, and Sephadex G-50 chromatography. The yield of growth factor is 5 mg per kg of pituitary. It is active in stimulating DNA synthesis in 3T3 cells at concentrations as low as 2 times 10-13 M with saturation at 1 times 10-10 M.     

The effect of serum albumin and the effect of cell concentration on the in vitro growth of mouse and rat lymphocytes.

The growth of mouse and rat T and B lymphocytes, activated by concanavalin A or lipopolysaccharide, is increased over growth in protein-free medium 5–20-fold by human or bovine serum albumin. The growth is dependent on the concentration of albumin. At a concentration of 2–4 mg/ml the growth rate is the same as in the presence of 10% fetal bovine serum. Of the other serum proteins (Cohn fractions) only human fraction VI supports growth somewhat while human fractions II–IV and bovine fraction VI do not support growth. The growth of mouse and rat lymphocytes is greatly suppressed if lymphocytes are cultured at high cell concentrations, and the growth-promoting ability of serum albumin cannot be detected under such conditions. The growth rate can be improved by daily adjustment of the pH, by daily refeeding, and by daily change of medium. The growth inhibitory activity can be removed largely by dialysis. It is concluded that the suppression of growth at high cell concentrations is caused by a combination of effects, i.e., a shift of pH, lack of nutrients, and accumulation of cellular metabolites.     

Inhibition and promotion by abscisic acid of growth in Spirodela

Summary The effect of different concentrations of abscisic acid on the growth of Spirodela oligorrhiza has been studied. Abscisic acid effectively permanently arrests growth at concentrations down to 10^-1 mg/l (one part per 10 million). Normal growth tends to be resumed at concentrations of 10^-2 and 10^-3 mg/l between 9 and 12 days after treatment. A concentration of 10^-8 mg/l, however, results in a significant increase in dry weight at both 9 and 12 days after introduction to the culture medium. It is suggested that the resumption of growth 12 days after treatment at those concentrations which inhibit growth up to 9 days, is due to a possible progressive inactivation of abscisic acid resulting in a lowering of its concentration to a level that is promotive. Sterile plants must be used since abscisic acid has shown no effect whatsoever in controlling growth of Saprolegnia, a water-mould of the Phycomycetes to which Spirodela apparently is the host.The authors gratefully acknowledge R. J. Reynolds Tobacco Company, North Carolina, U.S.A., for donating the abscisic acid.This research was supported by a grant to the senior author from the Council for Industrial and Scientific Research, Pretoria, S.A.     

The Role of Thiamine as a Factor for the Growth of Brochothrix thermosphacta

Thiamine enhances growth of the meat spoilage organism, Brochothrix thermosphacta. It fulfils some, but not all, of the yeast-extract requirement of this organism. Oxythiamine acts as a growth inhibitor at high concentrations and the inhibition is readily reversed by thiamine. The possible roles of thiamine in the metabolism of Br. thermosphacta are discussed.     

Glycerol fermentation of 1,3-propanediol by Clostridium butyricum. Measurement of product inhibition by use of a pH-auxostat

Summary The fermentation of glycerol to 1,3-propanediol, acetate, and butyrate by Clostridium butyricum was studied with respect to growth inhibition by the accumulating products. The clostridia were grown in a pH-auxostat culture at low cell density and product concentration and near maximum growth rate. The products were then added individually to the medium in increasing concentrations and the resulting depression of growth rate was used as a quantitative estimate of product inhibition. Under these conditions growth was totally inhibited at concentrations of 60 g/l for 1,3-propanediol, 27 g/l for acetic acid and 19 g/l for butyric acid at pH 6.5. Appreciable inhibition by glycerol was found only above a concentration of 80 g/l. In a pH-auxostat without added products but with high cell density as well as in batch cultures the product proportions were different. The 1,3-propanediol concentration may approach the value of complete inhibition while the concentrations of acetic and butyric acids remained below these values by at least one order of magnitude. It was therefore concluded that 1,3-propanediol is the first range inhibitor in this fermentation.     

Differences in growth factor sensitivity between individual 3T3 cells arise at high frequency: Possible relevance to cell senescence

At low serum concentrations (3% or less), individual Swiss 3T3 cells display marked heterogeneity in proliferative capacity. Here we show that this heterogeneity arises at extremely high frequency within a clone, often with sister cells showing considerable differences in capacity for further proliferation. The heterogeneity is unlikely to be due to genetic instability or mutation. Instead, it appears to reflect physiological differences between cells in their requirement for serum growth factors. It is suggested that these differences arise because cells are unable to sustain production, at low growth factor concentrations, of some rare component which is itself required for growth factor action. We believe that the generation of heterogeneity in 3T3 cells has much in common with the phenomenon of senescence in diploid cells.