The Effect of Inhibitory Substances on Cell Division

1950 ◽  
Vol s3-91 (15) ◽  
pp. 251-277
Author(s):  
A.F. W. HUGHES
Keyword(s):  
Cell Division ◽  
Phase Contrast ◽  
Tissue Cultures ◽  
Cell Divisions ◽  
High Concentrations ◽  
Chick Tissue ◽  
Very High ◽  
The Way

1. The effect of inhibitory substances upon cell divisions in chick tissue cultures has been, studied mainly by phase-contrast cine-micrography. 2. Fluoride and powerful -SH reactants prevent the entry of cells into prophase within I hour. 3. Aminopterin and colchicine are complete spindle inhibitors; they prevent its formation at the end of prophase, and can act on all metaphase cells in the culture. 4. The effect of fluoride, cyanide, and urethane on the spindle is much less drastic. 5. Most substances can retard nuclear reconstruction in telophase to some extent, but only chloracetophenone acts in the way already described for iodoacetamide. 6. Malonate appears to have no effect on mitosis, and fluoracetate only at very high concentrations. 7. Possible interpretations of some of these results are discussed.

Mitosis Is Not Required for Coordinated Changes in Iron Metabolism and High-Level Globin Gene Transcription in Erythropoietin-Responsive CD34+ Cells.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1776-1776
Author(s):  
Ajoy V. Bhupatiraju ◽  
Nicole M. Josleyn ◽  
Y. Terry Lee ◽  
Jeffery L. Miller
Keyword(s):  
Cell Division ◽  
Globin Gene ◽  
Beta Globin ◽  
Globin Mrna ◽  
Gamma Globin ◽  
Cell Divisions ◽  
Adult Human ◽  
Human Cd34 ◽  
Cd34 Cells ◽  
Very High

Abstract Within the erythroid lineage, erythropoietin (EPO) responsiveness is manifested by cell division and growth as well as specific changes in heme and globin production that ultimately result in the production of erythrocytes. However, the nascent relationship between EPO-associated mitosis and globin gene regulation has not been fully defined. In this study, cultured adult human CD34+ cells from peripheral blood were used to investigate early cellular responses to erythropoietin in the context of mitosis. Matched cultures were performed in replicate using human cells from at least two healthy adults. To detect mitosis, one million cells were labeled with 2uM carboxyfluorescein diacetate succinamidyl ester (CFSE). The CFSE-labeled cells were then cultured in the presence [EPO(+)] or absence [EPO(−)] of 4U/mL EPO, and analyzed using flow cytometry. No cell divisions were detected in either condition during the first 24 hours in culture, and multiple cell divisions were noted on subsequent culture days only in the EPO(+) cultures. Remarkably, dual-staining with CFSE and CD71 revealed a small (<1%) population of the undivided cells at 24 hours with very high surface levels of transferrin receptor [CD71(++++)] exclusively in the EPO(+) cultures. Further analysis of those rare EPO-responsive, pre-mitotic cells revealed DNA synthesis and entry into the cell cycle in 62.5±4.7% compared with 1.5±1.8% among the cells with lower CD71 expression. None expressed glycophorin A. Based upon their distinct phenotype, single cells were sorted into 96-well plates, with sorting confirmation by quantitative RT-PCR of GAPDH mRNA (20 copies/cell detection limit). Next, gamma- and beta-globin transcripts were amplified for comparison (4 separate experiments). Among the pre-mitotic, CD71(++++) population, 20 of 182 total sorted cells (11%) lacked detectable levels of gamma- and beta-globin mRNA. Only 1 of 182 cells (0.5%) expressed gamma-globin mRNA and no detectable beta globin mRNA. 138 of 182 cells (76%) expressed only beta-globin mRNA, and 23 of 182 cells (13%) expressed both globin mRNAs. The median and mean levels of gamma-globin mRNA among the 24 gamma(+) cells were 92 and 245±1015 copies/cell respectively. In contrast, the median and mean levels of beta-globin mRNA among the 161 beta(+) cells were 1624 and 3999±5892 copies/cell respectively. By comparison, the CD34+ cells with low levels of surface CD71 in either EPO(+) or EPO(−) cultures demonstrated detectable levels of gamma globin mRNA in only 7 of 145 sorted cells (5%), and beta-globin mRNA in 85 of 145 cells (59%), with total (gamma+beta) globin mRNA under 200 copies/cell in >90% of those cells. These novel results suggest that the frequency and levels of gamma-globin transcripts are quite low at the earliest stages of an EPO response among adult human CD34+ cells. However, it is clearly demonstrated that the cells can increase their capacity to import iron and transcribe beta-globin mRNA at very high levels prior to their first EPO-dependent cell division.

scholarly journals Rapid accumulation of mutations in growing mycelia of a hypervariable fungus Schizophyllum commune

2019 ◽  
Author(s):  
Aleksandra V. Bezmenova ◽  
Elena A. Zvyagina ◽  
Anna V. Fedotova ◽  
Artem S. Kasianov ◽  
Tatiana V. Neretina ◽  
...  
Keyword(s):  
Cell Division ◽  
Mutation Rate ◽  
De Novo ◽  
Schizophyllum Commune ◽  
Natural Populations ◽  
Germline Cell ◽  
Cell Divisions ◽  
Mutation Burden ◽  
Armillaria Gallica ◽  
Very High

AbstractThe number of mutations that occur per nucleotide per generation varies between species by several orders of magnitude. In multicellular eukaryotes, the per generation mutation rate depends both on the per cell division mutation rate and on the number of germline cell divisions per generation. In a range of species, from fungi to humans, the number of germline cell divisions is lower than that of somatic cells, reducing the mutation burden on the offspring. The basidiomycete Schizophyllum commune has the highest level of genetic polymorphism known among eukaryotes. In a previous study, it was also found to have a high per generation mutation rate, probably contributing to its high polymorphism. However, this rate has been measured only in a breeding experiment on Petri dishes, and it is unclear how this result translates to natural populations. Here, we used an experimental design that measures the rate of accumulation of de novo mutations in a linearly growing mycelium. We show that S. commune accumulates mutations at a uniform rate of 1.4·10−7 substitutions per nucleotide per meter of growth, which is 3 orders of magnitude higher than the corresponding rates in the oak Quercus robur and the fungus Armillaria gallica. This figure is consistent with the estimate obtained before, and suggests the lack of a dedicated germline in this system. If so, even a low per cell division mutation rate can translate into a very high per generation mutation rate when the number of cell divisions between consecutive meioses is large.

Stimulated Virus Production in Vinblastine and Cytochalasin-Induced Multinucleate Cells

1970 ◽  
Vol 28 ◽  
pp. 186-187
Author(s):  
Krishan Awtar
Keyword(s):  
Cell Division ◽  
Normal Cell ◽  
Virus Production ◽  
Multinucleate Cells ◽  
Daughter Cells ◽  
Cell Divisions ◽  
Nuclear Membranes ◽  
Division 2 ◽  
Vinblastine Sulfate

Exposure of cells to low sublethal but mitosis-arresting doses of vinblastine sulfate (Velban) results in the initial arrest of cells in mitosis followed by their subsequent return to an “interphase“-like stage. A large number of these cells reform their nuclear membranes and form large multimicronucleated cells, some containing as many as 25 or more micronuclei (1). Formation of large multinucleate cells is also caused by cytochalasin, by causing the fusion of daughter cells at the end of an otherwise .normal cell division (2). By the repetition of this process through subsequent cell divisions, large cells with 6 or more nuclei are formed.

The metachor as a characteristic of the association of electrolytes in aqueous solutions

1984 ◽  
Vol 49 (5) ◽  
pp. 1061-1078 ◽  
Author(s):  
Jiří Čeleda ◽  
Stanislav Škramovský
Keyword(s):  
Aqueous Solutions ◽  
Apparent Volume ◽  
Solutions Of Electrolytes ◽  
Molal Volumes ◽  
The Difference ◽  
High Concentrations ◽  
Experimental Values ◽  
Suitable Characteristic ◽  
Association Of Ions ◽  
Very High

Based on the earlier paper introducing a concept of the apparent parachor of a solute in the solution, we have eliminated in the present work algebraically the effect which is introduced into this quantity by the additivity of the apparent molal volumes. The difference remaining from the apparent parachor after substracting the contribution corresponding to the apparent volume ( for which the present authors suggest the name metachor) was evaluated from the experimental values of the surface tension of aqueous solutions for a set of 1,1-, 1,2- and 2,1-valent electrolytes. This difference showed to be independent of concentration up to the very high values of the order of units mol dm-3 but it was directly proportional to the number of the free charges (with a proportionality factor 5 ± 1 cm3 mol-1 identical for all studied electrolytes). The metachor can be, for this reason, a suitable characteristic for detection of the association of ions and formation of complexes in the solutions of electrolytes, up to high concentrations where other methods are failing.

scholarly journals Nitrification Process in a Nuclear Wastewater with High Load of Nitrogen, Uranium and Organic Matter under ORP Controlled

Water ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1607
Author(s):  
Mariano Venturini ◽  
Ariana Rossen ◽  
Patricia Silva Paulo
Keyword(s):  
Ammonia Oxidation ◽  
High Load ◽  
Nuclear Fuels ◽  
Monod Model ◽  
Nernst Equation ◽  
Nitrification Process ◽  
Real Wastewater ◽  
High Concentrations ◽  
Wastewater Samples ◽  
Very High

To produce nuclear fuels, it is necessary to convert uranium′s ore into UO2-ceramic grade, using several quantities of kerosene, methanol, nitric acid, ammonia, and, in low level, tributyl phosphate (TBP). Thus, the effluent generated by nuclear industries is one of the most toxic since it contains high concentrations of dangerous compounds. This paper explores biological parameters on real nuclear wastewater by the Monod model in an ORP controlled predicting the specific ammonia oxidation. Thermodynamic parameters were established using the Nernst equation to monitor Oxiders/Reductors relationship to obtain a correlation of these parameters to controlling and monitoring; that would allow technical operators to have better control of the nitrification process. The real nuclear effluent is formed by a mixture of two different lines of discharges, one composed of a high load of nitrogen, around 11,000 mg/L (N-NH4+-N-NO3−) and 600 mg/L Uranium, a second one, proceeds from uranium purification, containing TBP and COD that have to be removed. Bioprocesses were operated on real wastewater samples over 120 days under controlled ORP, as described by Nernst equations, which proved to be a robust tool to operate nitrification for larger periods with a very high load of nitrogen, uranium, and COD.

scholarly journals Halogenated Volatile Organic Compounds in Water Samples and Inorganic Elements Levels in Ores for Characterizing a High Anthropogenic Polluted Area in the Northern Latium Region (Italy)

2021 ◽  
Vol 18 (4) ◽  
pp. 1628
Author(s):  
Mario Vincenzo Russo ◽  
Ivan Notardonato ◽  
Alberto Rosada ◽  
Giuseppe Ianiri ◽  
Pasquale Avino
Keyword(s):  
Organic Compounds ◽  
Mean Value ◽  
Greenhouse Crops ◽  
River Waters ◽  
Liquid Liquid Extraction ◽  
Volatile Organic ◽  
High Concentrations ◽  
Very High ◽  
Inorganic Fraction

This paper shows a characterization of the organic and inorganic fraction of river waters (Tiber and Marta) and ores/soil samples collected in the Northern Latium region of Italy for evaluating the anthropogenic/natural source contribution to the environmental pollution of this area. For organic compounds, organochloride volatile compounds in Tiber and Marta rivers were analyzed by two different clean-up methods (i.e., liquid–liquid extraction and static headspace) followed by gas chromatography–electron capture detector (GC-ECD) analysis. The results show very high concentrations of bromoform (up to 1.82 and 3.2 µg L−1 in Tiber and Marta rivers, respectively), due to the presence of greenhouse crops, and of chloroform and tetrachloroethene, due to the presence of handicrafts installations. For the qualitative and quantitative assessment of the inorganic fraction, it is highlighted the use of a nuclear analytical method, instrumental neutron activation analysis, which allows having more information as possible from the sample without performing any chemical-physical pretreatment. The results have evidenced high levels of mercury (mean value 88.6 µg g−1), antimony (77.7 µg g−1), strontium (12,039 µg g−1) and zinc (103 µg g−1), whereas rare earth elements show levels similar to the literature data. Particular consideration is drawn for arsenic (414 µg g−1): the levels found in this paper (ranging between 1 and 5100 µg g−1) explain the high content of such element (as arsenates) in the aquifer, a big issue in this area.

THE SELECTIVE PROTEOLYTIC ENZYME INHIBITORY ACTION OF BENZETHONIUM CHLORIDE

1960 ◽  
Vol 38 (1) ◽  
pp. 25-32 ◽  
Author(s):  
Ivan T. Beck ◽  
E. Pinter ◽  
R. D. McKenna ◽  
H. Griff
Keyword(s):  
Subcutaneous Injection ◽  
Proteolytic Enzyme ◽  
Inhibitory Action ◽  
Hemorrhagic Pancreatitis ◽  
Benzethonium Chloride ◽  
Tryptic Activity ◽  
High Concentrations ◽  
Acute Hemorrhagic Pancreatitis ◽  
Very High

Acute hemorrhagic pancreatitis in humans is thought to be perpetuated by the autolytic processes catalyzed by trypsin and lipase. This study is an integral part of our search for trypsin and lipase inhibitors to be used in the treatment of this disease.Benzethonium chloride was found to inhibit tryptic activity in vitro. The proteolytic activity of rabbit's serum was inhibited, and the inhibition was most pronounced 6 to 12 hours after the subcutaneous injection of the compound. Fibrinolysin was also inhibited in vitro but benzethonium chloride had no inhibitory action on chymotrypsin, pepsin, or lipase.Serum proteins in vitro were precipitated only with very high concentrations of the compound. No significant protein changes were observed in sera of rabbits after the subcutaneous injection of the compound.

scholarly journals Arginine Inhibits Serpins

2007 ◽  
Vol 13 (2) ◽  
pp. 213-218
Author(s):  
Thomas W. Stief
Keyword(s):  
Esterase Activity ◽  
Serine Proteases ◽  
Biological Fluid ◽  
Microtiter Plate ◽  
Human Albumin ◽  
Amidolytic Activity ◽  
Peptide Substrates ◽  
Plate Reader ◽  
High Concentrations ◽  
Very High

Serine protease inactivators (serpins) are important regulators in biochemistry. Often it is necessary to block the serpin action, that is, to stabilize the sample. The guanidine group of arginine is the ligand for the active center pocket of many serine proteases. Arginine or guanidine inhibits serine proteases, and arginine belongs to the reactive P1-P1' center of many serpins. The plasmatic antithrombin, antiplasmin, or anti-C1-esterase activity was determined: A total of 20 µL of pooled normal plasma or 7% human albumin was added to 100 µL of 0—2.67 M arginine, pH 8.6, 10 µL of 26 mIU/mL thrombin in 7% human albumin, and 30 µL of 1.7 mM CHG-Ala-Arg-pNA (37°C). ΔA at 405 nm was determined, by using a microtiter plate reader. Thrombin was substituted by plasmin or C1-esterase, and the chromogenic peptide substrates <Glu-Phe-Lys-pNA or MeOC-Lys(eCBO)-Gly-Arg-pNA, respectively, were used. The IC50 of arginine against plasmatic antithrombin activity is 580 mM; the IC 25 is 440 mM. The IC25 of arginine against plasmatic α 2-antiplasmin or C1-inactivator is 1650 mM. The amidolytic activity of thrombin, plasmin, and C1-esterase is inhibited similarly by arginine: the IC50 for arginine against the amidolytic activity of these proteases is about 400 mM. Arginine at very high concentrations inhibits serpins. This is important, if stabilization of a biological fluid is a prerequisite for valid activities of serine proteases. In addition, these high concentrations of arginine might be a new gentle principle to inhibit pathogens that need serpins for their pathophysiology.

scholarly journals THE SUBMICROSCOPIC ORGANIZATION OF AXON MATERIAL ISOLATED FROM MYELIN NERVE FIBERS

1953 ◽  
Vol 98 (3) ◽  
pp. 269-276 ◽  
Author(s):  
E. De Robertis ◽  
C. M. Franchi
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Phase Contrast ◽  
Nerve Fibers ◽  
Myelinated Nerve ◽  
Myelinated Nerve Fibers ◽  
Dense Granules ◽  
Small Clusters ◽  
Membranous Material ◽  
The Way

A technique has been developed for the extrusion of axon material from myelinated nerve fibers. This material is then compressed and prepared for observation with the electron microscope. All the stages of preparation and purification of the axon material can be checked microscopically and in the present paper they are illustrated with phase contrast photomicrographs. Observation with the electron microscope of the compressed axons showed the presence of the following components: granules, fibrils, and a membranous material. Only the larger granules could be seen with the ordinary microscope. A considerable number of dense granules were observed. Of these the largest resemble typical mitochondria of 250 mµ by 900 mµ. In addition rows or small clusters of dense granules ranging in diameter from 250 to 90 mµ were present. In several specimens fragments of a membrane 120 to 140 A thick and intimately connected with the axon were found. The entire axon appeared to be constituted of a large bundle of parallel tightly packed fibrils among which the granules are interspersed. The fibrils are of indefinite length and generally smooth. They are rather labile structures, less resistant in the rat than in the toad nerve. They varied between 100 and 400 A in diameter and in some cases disintegrated into very fine filaments (less than 100 A thick). The significance is discussed of the submicroscopic structures revealed by electron microscopy of the material prepared in the way described.

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