scholarly journals Maternal serum supplementation in culture medium benefits maturation of immature human oocytes

2001 ◽  
Vol 3 (2) ◽  
pp. 112-116 ◽  
Author(s):  
AL Mikkelsen ◽  
E Høst ◽  
J Blaabjerg ◽  
S Lindenberg
Author(s):  
Awtar Krishan ◽  
Dora Hsu

Cells exposed to antitumor plant alkaloids, vinblastine and vincristine sulfate have large proteinacious crystals and complexes of ribosomes, helical polyribosomes and electron-dense granular material (ribosomal complexes) in their cytoplasm, Binding of H3-colchicine by the in vivo crystals shows that they contain microtubular proteins. Association of ribosomal complexes with the crystals suggests that these structures may be interrelated.In the present study cultured human leukemic lymphoblasts (CCRF-CEM), were incubated with protein and RNA-synthesis inhibitors, p. fluorophenylalanine, puromycin, cycloheximide or actinomycin-D before the addition of crystal-inducing doses of vinblastine to the culture medium. None of these compounds could completely prevent the formation of the ribosomal complexes or the crystals. However, in cells pre-incubated with puromycin, cycloheximide, or actinomycin-D, a reduction in the number and size of the ribosomal complexes was seen. Large helical polyribosomes were absent in the ribosomal complexes of cells treated with puromycin, while in cells exposed to cycloheximide, there was an apparent reduction in the number of ribosomes associated with the ribosomal complexes (Fig. 2).


Author(s):  
C. Wiencke ◽  
A. Lauchli

Osmoregulatory mechanisms in algae were investigated mainly from a physiological point of view (KAUSS 1977, HELLEBUST 1976). In Porphyra two osmotic agents, i. e. floridoside/isofloridoside (KAUSS 1968) and certain ions, such as K+ and Na+(EPPLEY et al. 1960) are considered for osmotic balance. Accumulations of ions (particularly Na+) in the cytoplasm during osmotic adaptation is improbable, because the activity of enzymes is generally inhibited by high ionic concentrations (FLOWERS et al. 1977).The cellular organization of Porphyra was studied with special emphasis on the development of the vacuolar system under different hyperosmotic conditions. Porphyra was cultivated at various strengths of the culture medium ASP 12 (PROVASOLI 1961) ranging from normal to 6 times concentrated (6x) culture medium. Por electron microscopy freeze fracturing was used (specimens fixed in 2% glutaraldehyde and incubated in 30% glycerol, preparation in a BALZERS BA 360 M apparatus), because chemical fixation gave poor results.


Author(s):  
Etienne de Harven ◽  
Nina Lampen

Samples of heparinized blood, or bone marrow aspirates, or cell suspensions prepared from biopsied tissues (nodes, spleen, etc. ) are routinely prepared, after Ficoll-Hypaque concentration of the mononuclear leucocytes, for scanning electron microscopy. One drop of the cell suspension is placed in a moist chamber on a poly-l-lysine pretreated plastic coverslip (Mazia et al., J. Cell Biol. 66:198-199, 1975) and fifteen minutes allowed for cell attachment. Fixation, started in 2. 5% glutaraldehyde in culture medium at room temperature for 30 minutes, is continued in the same fixative at 4°C overnight or longer. Ethanol dehydration is immediately followed by drying at the critical point of CO2 or of Freon 13. An efficient alternative method for ethanol dehydrated cells is to dry the cells at low temperature (-75°C) under vacuum (10-2 Torr) for 30 minutes in an Edwards-Pearse freeze-dryer (de Harven et al., SEM/IITRI/1977, 519-524). This is preceded by fast quenching in supercooled ethanol (between -90 and -100°C).


Author(s):  
W. A. Chiou ◽  
N. Kohyama ◽  
B. Little ◽  
P. Wagner ◽  
M. Meshii

The corrosion of copper and copper alloys in a marine environment is of great concern because of their widespread use in heat exchangers and steam condensers in which natural seawater is the coolant. It has become increasingly evident that microorganisms play an important role in the corrosion of a number of metals and alloys under a variety of environments. For the past 15 years the use of SEM has proven to be useful in studying biofilms and spatial relationships between bacteria and localized corrosion of metals. Little information, however, has been obtained using TEM capitalizing on its higher spacial resolution and the transmission observation of interfaces. The research presented herein is the first step of this new approach in studying the corrosion with biological influence in pure copper.Commercially produced copper (Cu, 99%) foils of approximately 120 μm thick exposed to a copper-tolerant marine bacterium, Oceanospirillum, and an abiotic culture medium were subsampled (1 cm × 1 cm) for this study along with unexposed control samples.


Author(s):  
Jaang J. Wang ◽  
Cheng C. Chen ◽  
Men F. Shaio ◽  
Chia T. Liu ◽  
Chung S. Lee ◽  
...  

The involvement of nucleus in the maturation processes of Dengue-2 virus in a mosquito cell line, C6/36 cells, has been identified by the electron microscopy and immunocytochemistry. The C6/36 cells were obtained from ATCC and maintained in MEM culture medium containing 10% fetal bovine serum at 28°C. The cell suspensions or cells grown on teflon-coated coverslips were infected with Dengue-2 virus (107/ml) for various time periods of 2 hours, 3, 6, 8, and 10 days. The cells were then fixed in buffered 1.5% glutaraldehyde, and washed in acetone before immunolabeled with monoclonal antibody. An indirect immunocytochemical labeling method of avidin-biotin complex (ABC) conjugated with peroxidase or gold particles (20 nm in diameter) and a flat embedding technique were used to localize the virus particles.At early stages of infections (before 3 days), there were no virion particles detected. After 6 days and on of infections, cytopathic effect (CPE) was observed and showed positive immuno-peroxidase reactions under the light and electron microscopies.


Author(s):  
Douglas R. Keene ◽  
B. Kerry Maddox ◽  
Marie B. Spurgin ◽  
Lynn Y. Sakai ◽  
Robert W. Glanville

A mouse monoclonal antibody was used to identify beaded aggregates found in guanidine extracts of human amnion as assemblies of fibrillin molecules. These aggregates were also shown to be a major component of extracellular matrix microfibrils. We further demonstrated that the periodicity of these aggregates can be increased when subjected to mechanical stress.Human amnion was extracted with guanidine and the extracted material purified using ion exchange and molecular sieve chromatography. A high molecular weight fraction was precipitated by dialyzing against dilute acetic acid. Part of the precipitate was suspended in 0.2 M ammonium bicarbonate buffer and rotary shadowed. A second portion was resuspended in culture medium containing antibody which recognizes matrix microfibrils, diluted 1:5 in ammonium bicarbonate and reacted for 120 minutes at room temperature. Antibody labeled precipitate was washed by repeated pelleting and resuspension in buffer and then incubated in Janssen GAM 5 nm gold conjugate for 60 minutes at room temperature.


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