The effects of osmotic pressure changes on the germination of Dictyostelium discoideum spores

1977 ◽  
Vol 23 (9) ◽  
pp. 1170-1177 ◽  
Author(s):  
David A. Cotter
Keyword(s):  
Ethylene Glycol ◽  
Dictyostelium Discoideum ◽  
Room Temperature ◽  
Thermal Inactivation ◽  
Sucrose Concentration ◽  
Lag Phase ◽  
Activation Process ◽  
Sucrose Solutions ◽  
Spore Population ◽  
Pressure Changes

Polyalcohols such as ethylene glycol and glycerol at 3 M penetrate and activate spores of Dictyostelium discoideum incubated at room temperature. Higher concentrations of ethylene glycol result in lysis upon suspension of spores in dilute phosphate buffer. Erythritol and arabitol at 3 M do not penetrate or activate D. discoideum spores.Air-dried spores or those incubated in 2 M sucrose solutions are not activated with the usual heat treatment of 45 °C for 30 min. The plasmolyzed spores are activated at temperatures above 45 °C when heated in the presence of 2 M sucrose for 30 min. The temperature for maximum activation and the temperature for thermal inactivation of spores are raised 7–10 °C in high sucrose concentrations. Long-term incubation of heat-activated spores in 2 M sucrose solutions does not result in a return to dormancy.Moderate sucrose concentrations near 0.2 M do not block the heat-induced activation process but must be removed from the spore population to prevent a return to dormancy within 6 h. Other polyhydric compounds at 0.25 M concentration also cause spore deactivation within 6 h of room temperature incubation. Oxygen uptake of spores undergoing deactivation in 0.18 M sucrose is inhibited as compared to control levels. Moderate concentrations of sucrose do not block the early events of postactivation lag and the spores accumulate at the end of the lag phase. The longer the spores remain unswollen at the end of the postactivation lag phase, the greater the percentage of spores which return to dormancy. The effects of moderate sucrose concentration (lowered water activity) are not duplicated by the same quantity of Ficoll, indicating that the colligative properties of the sucrose solutions are responsible for deactivation.

68 OSMOTIC RESPONSES OF EQUINE EMBRYOS WITH AND WITHOUT CAPSULES TO CRYOPROTECTANTS

2011 ◽  
Vol 23 (1) ◽  
pp. 139 ◽  
Author(s):  
J. P. Barfield ◽  
S. P. Leibo ◽  
P. M. McCue ◽  
G. E. Seidel
Keyword(s):  
Ethylene Glycol ◽  
Room Temperature ◽  
Regression Analyses ◽  
Initial Volume ◽  
Original Volume ◽  
Sucrose Solutions ◽  
Percentage Volume ◽  
Size And Morphology ◽  
Volume Range ◽  
Volume Decrease

Equine embryos become more difficult to cryopreserve as they get larger, which may be due in part to the presence of the capsule. We hypothesized that presence of the capsule will alter osmotic responses of equine embryos. Embryos were collected from mares of light horse breeds. Ovulation was confirmed by daily transrectal ultrasonography, and uterine flushes were performed 7 or 8 days post-ovulation. At collection, embryos were evaluated for size and morphology and graded on a scale of 1 to 5, 1 being excellent and 5 being degenerate/dead. Only embryos of grades 1 or 2 were used. Boyle Van’t Hoff plots were created for embryos <300 (n = 2), 300–600 (n = 5), or >600 μm (n = 2) in diameter from data collected after incubating embryos in a series of increasingly concentrated sucrose solutions of 250, 500, 1000, and 2000 mOsm in Syngro® (275 mOsm) at room temperature (27–30°C). Embryo diameters were measured every minute for at least 60 min. Volumes were calculated, and relative volumes calculated against that were measured immediately after the embryo was flushed from the uterus (100%) while the embryo was in Syngro®. Regression analyses were used to estimate the y-intercept from which the percentage volume that is osmotically active water was calculated [91.9 for embryos < 300 μm (P = 0.287), 85.7 for 300–600 μm (P = 0.0005), and 80.0% for >600 μm (P = 0.003)]. To evaluate osmotic responses of equine embryos to 3 cryoprotectants, Day 7 embryos between 300 and 600 μm with or without a capsule were transferred to 5 mL of 1.5 M methanol (n = 3 with capsule, n = 2 without), 1.5 M glycerol (n = 5 with, n = 2 without), or 0.75 M ethylene glycol + 0.75 M methanol (n = 6 with, n = 3 without) in Syngro®. Diameters were measured every min for the first 30 min and every 5 min thereafter. Upon removal of the capsule and before exposure to cryoprotectants, embryos lost and did not recover on average 22% of their initial volume (range 9 to 30%) after 15 min of incubation in Syngro® at room temperature (27 to 30°C). When incubated in 1.5 M methanol, embryo volume increased ∼10%, and then decreased to ∼95% original volume after 1 h incubation for embryos that had intact capsules; embryos without capsules remained ∼5% larger than initial volume throughout incubation. When incubated in 1.5 M glycerol, embryos with capsules decreased to ∼40% within 2 min and did not recover volume during incubation; embryos without capsules decreased to ∼75% of original volume gradually over 60 min. The volume decrease of embryos with a capsule is similar to previous reports for embryos of similar size in 1.5 M glycerol. Embryos incubated in 0.75 M ethylene glycol + 0.75 M methanol without a capsule initially lost ∼15% of their volume and remained at ∼87% of initial volume for the duration of the experiment; embryos with capsules lost nearly 30% of their original volume but then recovered volume to ∼92% of original volume by the end of 1 h. These experiments suggest that the equine capsule may impede movement of glycerol and, to a lesser extent, ethylene glycol into the embryo.

scholarly journals Synthesis of Network Polymers by Means of Addition Reactions of Multifunctional-Amine and Poly(ethylene glycol) Diglycidyl Ether or Diacrylate Compounds

Polymers ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 2047
Author(s):  
Naofumi Naga ◽  
Mitsusuke Sato ◽  
Kensuke Mori ◽  
Hassan Nageh ◽  
Tamaki Nakano
Keyword(s):  
Ethylene Glycol ◽  
Room Temperature ◽  
Addition Reaction ◽  
Monomer Concentration ◽  
Diglycidyl Ether ◽  
Porous Polymers ◽  
Addition Reactions ◽  
Poly Ethylene Glycol ◽  
Network Polymers ◽  
Poly Ethylene

Addition reactions of multi-functional amine, polyethylene imine (PEI) or diethylenetriamine (DETA), and poly(ethylene glycol) diglycidyl ether (PEGDE) or poly(ethylene glycol) diacrylate (PEGDA), have been investigated to obtain network polymers in H2O, dimethyl sulfoxide (DMSO), and ethanol (EtOH). Ring opening addition reaction of the multi-functional amine and PEGDE in H2O at room temperature or in DMSO at 90 °C using triphenylphosphine as a catalyst yielded gels. Aza-Michael addition reaction of the multi-functional amine and PEGDA in DMSO or EtOH at room temperature also yielded corresponding gels. Compression test of the gels obtained with PEI showed higher Young’s modulus than those with DETA. The reactions of the multi-functional amine and low molecular weight PEGDA in EtOH under the specific conditions yielded porous polymers induced by phase separation during the network formation. The morphology of the porous polymers could be controlled by the reaction conditions, especially monomer concentration and feed ratio of the multi-functional amine to PEGDA of the reaction system. The porous structure was formed by connected spheres or a co-continuous monolithic structure. The porous polymers were unbreakable by compression, and their Young’s modulus increased with the increase in the monomer concentration of the reaction systems. The porous polymers absorbed various solvents derived from high affinity between the polyethylene glycol units in the network structure and the solvents.

Characterization and catalytic properties of a saponite clay modified by acid activation

Clay Minerals ◽  
1997 ◽  
Vol 32 (4) ◽  
pp. 633-643 ◽  
Author(s):  
F. Kooli ◽  
W. Jones
Keyword(s):  
Infrared Spectroscopy ◽  
Room Temperature ◽  
Textural Properties ◽  
Acid Sites ◽  
Single Type ◽  
Activation Process ◽  
Acid Activation ◽  
Strongly Correlated ◽  
Lewis Site ◽  
Adsorbed Pyridine

AbstractA natural saponite was acid activated at room temperature or 90°C with different acid/clay ratios and the products were characterized by powder X-ray diffraction, infrared spectroscopy and thermogravimetry. The leaching of Mg from the octahedral sheets is enhanced by an increase in the acid/clay ratio and by an increase in temperature of activation. Textural properties are reported, and it appears that they are strongly correlated to the presence of a noncrystalline silica phase which is formed during the acid activation process. The desorption of cyclohexylamine indicates that for samples activated at 90°C the number of acid sites in the acidactivated saponites decreases following severe acid treatment. Infrared spectroscopy of adsorbed pyridine on samples after calcination at 500°C suggests that acid activation at 90°C produces a single type of Bronsted site but two types of Lewis sites whereas activation at room temperature results in only one type of Lewis site in addition to a Brønsted site. The two Lewis sites are suggested to originate from residual Al in the clay structure and to AI exsolved from the layers during activation. The dehydration of pentan-1-ol has been used as a further probe to measure acidity by monitoring the degree of conversion and selectivity for the different samples.

Characterisation of an epithelium-like layer of cells in the multicellular Dictyostelium discoideum slug

1993 ◽  
Vol 105 (1) ◽  
pp. 243-253
Author(s):  
M. Fuchs ◽  
M.K. Jones ◽  
K.L. Williams
Keyword(s):  
Dictyostelium Discoideum ◽  
Intercellular Space ◽  
Room Temperature ◽  
Cell Layer ◽  
Freeze Substitution ◽  
Specific Cell ◽  
Peripheral Layer ◽  
Apical Membranes ◽  
Multicellular Organisms ◽  
Peripheral Cells

Ultrarapid freezing (RF) followed by freeze-substitution (FS) provide superior preservation of the Dictyostelium discoideum multicellular slug tissue over conventional methods of chemical fixation at room temperature. The peripheral cells of slugs prepared by RF and FS form a tight layer of flattened cells. This cell layer resembles epithelia of other multicellular organisms in that it has close junctional contact between cells associated with the extracellular matrix (ECM, slime sheath). This is the first report that clearly demonstrates the existence of such peripheral cellular specialisation in this otherwise well-studied model system. Junctional contacts between adjacent cells mean that there is no intercellular space evident between apical membranes of apposing cells, and basally the intermembraneous space between peripheral cells is less than 10 nm. By contrast, the intercellular space between internal cells is approximately 10–25 nm. The shape of the peripheral cells varies with their location around the slug. In the posterior prespore zone, the peripheral cells are squamous and exhibit polarity along their antero-posterior axis. In the anterior prestalk zone, peripheral cells are less flattened, project irregular filipodia between internal cells, and are polarised along their apical-basal axis. Colloidal gold immunocytochemistry with the markers MUD1, MUD50 and MUD62 demonstrated that the peripheral layer is formed of prestalk cells in the anterior region and ventrum, and mostly prespore cells along the dorsum. Thus, the peripheral layer, while having specific cell classes in different regions, is not differentiation-specific. Rather, it appears that the structure of these epithelium-like cells is influenced by interaction with molecules of the ECM (sheath).

Kinetics of endosomal pH evolution in Dictyostelium discoideum amoebae. Study by fluorescence spectroscopy

1993 ◽  
Vol 105 (3) ◽  
pp. 861-866 ◽  
Author(s):  
L. Aubry ◽  
G. Klein ◽  
J.L. Martiel ◽  
M. Satre
Keyword(s):  
Dictyostelium Discoideum ◽  
Fluid Phase ◽  
Lag Phase ◽  
Nutritive Medium ◽  
Chase Period ◽  
Endosomal Acidification ◽  
Fluid Phase Endocytosis ◽  
Endosomal Ph ◽  
Efflux Kinetics ◽  
Secondary Lysosomes

The evolution of endo-lysosomal pH in Dictyostelium discoideum amoebae was examined during fluid-phase endocytosis. Pulse-chase experiments were conducted in nutritive medium or in non-nutritive medium using fluorescein labelled dextran (FITC-dextran) as fluid-phase marker and pH probe. In both conditions, efflux kinetics were characterized by an extended lag phase lasting for 45–60 min and corresponding to intracellular transit of FITC-dextran cohort. During the chase period, endosomal pH decreased during approximately 20 min from extracellular pH down to pH 4.6-5.0, then, it increased within the next 20–40 min to reach pH 6.0-6.2. It was only at this stage that FITC-dextran was released back into the medium with pseudo first-order kinetics. A vacuolar H(+)-ATPase is involved in endosomal acidification as the acidification process was markedly reduced in mutant strain HGR8, partially defective in vacuolar H(+)-ATPase and in parent type strain AX2 by bafilomycin A1, a selective inhibitor of this enzyme. Our data suggest that endocytic cargo is channeled from endosomes to secondary lysosomes that are actively linked to the plasma membrane via recycling vesicles.

scholarly journals Ti-V-C-Based Alloy with a FCC Lattice Structure for Hydrogen Storage

Molecules ◽  
2019 ◽  
Vol 24 (3) ◽  
pp. 552
Author(s):  
Bo Li ◽  
Liqing He ◽  
Jianding Li ◽  
Hai-Wen Li ◽  
Zhouguang Lu ◽  
...  
Keyword(s):  
Hydrogen Storage ◽  
Room Temperature ◽  
Lattice Structure ◽  
Activation Process ◽  
Face Centered Cubic ◽  
Hydrogen Storage Materials ◽  
Fcc Lattice ◽  
Bcc Structure ◽  
Face Centered ◽  
Fcc Structure

Here we report a Ti50V50-10 wt.% C alloy with a unique lattice and microstructure for hydrogen storage development. Different from a traditionally synthesized Ti50V50 alloy prepared by a melting method and having a body-centered cubic (BCC) structure, this Ti50V50-C alloy synthesized by a mechanical alloying method is with a face-centered cubic (FCC) structure (space group: Fm-3m No. 225). The crystalline size is 60 nm. This alloy may directly absorb hydrogen near room temperature without any activation process. Mechanisms of the good kinetics from lattice and microstructure aspects were discussed. Findings reported here may indicate a new possibility in the development of future hydrogen storage materials.

scholarly journals Teaching a biology laboratory course using Dictyostelium

2019 ◽  
Vol 63 (8-9-10) ◽  
pp. 551-561
Author(s):  
David A. Knecht ◽  
Kate M. Cooper ◽  
Jonathan E. Moore
Keyword(s):  
Dictyostelium Discoideum ◽  
Cell Biology ◽  
Room Temperature ◽  
Model System ◽  
Educational Settings ◽  
Biology Teaching ◽  
Laboratory Course ◽  
Cellular Processes ◽  
Laboratory Courses ◽  
Biology Laboratory

The Dictyostelium discoideum model system is a powerful tool for undergraduate cell biology teaching laboratories. The cells are biologically safe, grow at room temperature and it is easy to experimentally induce, observe, and perturb a breadth of cellular processes making the system amenable to many teaching lab situations and goals. Here we outline the advantages of Dictyostelium, discuss laboratory courses we teach in three very different educational settings, and provide tips for both the novice and experienced Dictyostelium researcher. With this article and the extensive sets of protocols and tools referenced here, implementing these labs, or parts of them, will be relatively straightforward for any instructor.

Thermal Inactivation of Borrelia burgdorferi, the Cause of Lyme Disease

1990 ◽  
Vol 53 (4) ◽  
pp. 296-299 ◽  
Author(s):  
SI K. LEE ◽  
AHMED E. YOUSEF ◽  
ELMER H. MARTH
Keyword(s):  
Borrelia Burgdorferi ◽  
Thermal Inactivation ◽  
Treatment Time ◽  
Treatment Temperature ◽  
Most Probable Number ◽  
Lag Phase ◽  
Probable Number ◽  
Heat Treated ◽  
Thermal Death ◽  
D Values

Borrelia burgdorferi strain EBNI was cultivated in BSK-II medium at 34°C, then cultures at different physiological states were heat-treated at temperatures in the range of 50 to 70°C. Numbers of survivors were estimated by the Most Probable Number technique. Log MPN was plotted against treatment time, and resulting survivor curves were linear. Estimated D-values for cultures incubated at 34°C for 7 d before heat-treatment were 5.5, 4.3, 2.7, .47, and .14 min at 50, 55, 60, 65, and 70°C, respectively. Spirochetes in the lag phase had greater resistance to heat than those in the stationary phase, with the latter being more resistant to heat than spirochetes in the same phase of growth but refrigerated at 4°C for 3 d. D-values for B. burgdorferi are generally less at 50°C, and greater at 70°C than those reported for other nonsporeforming pathogens. When log10 MPN was plotted against treatment temperature, two linear segments for each thermal death curve were obtained. Our data show the spirochete had higher z-values than most nonsporeforming pathogens. The pH of the medium, in the range of 5.0 to 7.6, did not affect resistance of B. burgdorferi to heat.

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