Concentration of enteroviruses from large volumes of turbid estuary water

1977 ◽  
Vol 23 (6) ◽  
pp. 770-778 ◽  
Author(s):  
Mark D. Sobsey ◽  
Charles P. Gerba ◽  
Craig Wallis ◽  
Joseph L. Melnick
Keyword(s):  
Ferric Chloride ◽  
Aluminum Chloride ◽  
Fetal Calf Serum ◽  
Sea Water ◽  
Calf Serum ◽  
Glycine Buffer ◽  
In Series ◽  
Overall Efficiency

A method is described for the efficient concentration of viruses from large volumes of highly turbid estuary water. Virus in acidified sea water in the presence of aluminum chloride is adsorbed to a 10-in. (about 25.4 cm) fibreglass depth cartridge and 2- and 0.65-μm epoxy-fibreglass filters in series. This filter series is capable of efficiently adsorbing enteroviruses from 50 U.S. gallons (about 190 ℓ) of estuary water of varying salinity and turbidity. Adsorbed viruses were eluted from the filters with glycine buffer (pH 11.5) and the eluate reconcentrated by using a precipitate formed by the addition of ferric chloride. Viruses were eluted from this precipitate with fetal calf serum. Using this procedure, four different enteroviruses in 50 gallons (about 190 ℓ) of estuary water were concentrated 9 000- to 12 000-fold with an overall efficiency of 41%.

scholarly journals Cryopreservation of enucleated human neutrophils (PMN cytoplasts)

Blood ◽  
1984 ◽  
Vol 63 (1) ◽  
pp. 234-237 ◽  
Author(s):  
AA Voetman ◽  
AA Bot ◽  
D Roos
Keyword(s):  
Hydrogen Peroxide ◽  
Staphylococcus Aureus ◽  
Alkaline Phosphatase ◽  
Fetal Calf Serum ◽  
Unit Area ◽  
Calf Serum ◽  
Human Neutrophils ◽  
In Series ◽  
And Storage

Abstract Previously, we have shown that enucleated human neutrophils (PMN cytoplasts), when activated by particulate or fluid stimuli, generate superoxide and hydrogen peroxide at rates comparable (per unit area of plasma membrane) to those observed with intact neutrophils. Moreover, PMN cytoplasts also ingest and, to a certain extent, kill bacteria. We now report that PMN cytoplasts can be cryopreserved with maintenance of their functional activity. The PMN cytoplasts were frozen in a medium with 10% (v/v) fetal calf serum and 10% (v/v) dimethyl sulfoxide, and stored at -70 degrees C. After thawing and washing, the recovery was 75%. The content of alkaline phosphatase and lactate dehydrogenase, the consumption of oxygen and generation of hydrogen peroxide, and the rate of phagocytosis of Staphylococcus aureus bacteria was the same for fresh and cryopreserved PMN cytoplasts. Identical values were obtained after preservation in liquid nitrogen. These results open possibilities to store neutrophil material, allowing longitudinal follow-up of patients, comparative studies between different patients, exchange of material between laboratories, and storage of reference material for experiments in series.

scholarly journals Cryopreservation of enucleated human neutrophils (PMN cytoplasts)

Blood ◽  
1984 ◽  
Vol 63 (1) ◽  
pp. 234-237
Author(s):  
AA Voetman ◽  
AA Bot ◽  
D Roos
Keyword(s):  
Hydrogen Peroxide ◽  
Staphylococcus Aureus ◽  
Alkaline Phosphatase ◽  
Fetal Calf Serum ◽  
Unit Area ◽  
Calf Serum ◽  
Human Neutrophils ◽  
In Series ◽  
And Storage

Previously, we have shown that enucleated human neutrophils (PMN cytoplasts), when activated by particulate or fluid stimuli, generate superoxide and hydrogen peroxide at rates comparable (per unit area of plasma membrane) to those observed with intact neutrophils. Moreover, PMN cytoplasts also ingest and, to a certain extent, kill bacteria. We now report that PMN cytoplasts can be cryopreserved with maintenance of their functional activity. The PMN cytoplasts were frozen in a medium with 10% (v/v) fetal calf serum and 10% (v/v) dimethyl sulfoxide, and stored at -70 degrees C. After thawing and washing, the recovery was 75%. The content of alkaline phosphatase and lactate dehydrogenase, the consumption of oxygen and generation of hydrogen peroxide, and the rate of phagocytosis of Staphylococcus aureus bacteria was the same for fresh and cryopreserved PMN cytoplasts. Identical values were obtained after preservation in liquid nitrogen. These results open possibilities to store neutrophil material, allowing longitudinal follow-up of patients, comparative studies between different patients, exchange of material between laboratories, and storage of reference material for experiments in series.

Banking of embryos of mutated, paralytic tremor rabbit by means of vitrification

2005 ◽  
Vol 39 (3) ◽  
pp. 284-289 ◽  
Author(s):  
Krzysztof Papis ◽  
Joanna Sypecka ◽  
Maciej Korwin-Kossakowski ◽  
Elżbieta Wenta-Muchalska ◽  
Barbara Bilska
Keyword(s):  
Germ Plasm ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Blastocyst Stage ◽  
Laboratory Animals ◽  
Real Possibility ◽  
The Central Nervous System ◽  
Recessive Mutant ◽  
Overall Efficiency

Cryopreservation enables banking of embryos for future use in medicine and in animal breeding. It also enables protection of germ plasm of endangered species and unique strains or lanes of laboratory animals. This paper describes an example of employing a vitrification method for banking of embryos of a unique lane of rabbit. The paralytic tremor ( pt) rabbit is an X-linked recessive mutant lane of the Chinchilla breed characterized by hypomyelination of the central nervous system. In order to obtain a sufficient number of embryos, pt females were subjected to superovulation and surgical embryo collection. All suitable embryos were vitrified in 0.25 mL insemination straws in a modified EFS vitrification solution comprised of ethylene glycol (40%), Ficoll 70 (18%) and sucrose (0.3 M) in Hepes buffered TCM medium containing 20% fetal calf serum. In order to assess the efficiency of the vitrification procedure, a representative portion of vitrified embryos was warmed after a period of storage. Warmed embryos were subjected to in vitro culture for 72 h or were transferred to the uterus of synchronized recipients. The majority of the 141 warmed embryos survived vitrification and 100/141 (71%) developed to the blastocyst stage. Moreover, out of an additional 34 warmed embryos transferred to four recipients, eight (23.5%) developed to term and seven live pups were born. Six of the rabbit pups exhibited paralytic tremor symptoms typical for the pt lane. Although the overall efficiency of the vitrification method was lower compared with the effects usually achieved for 'healthy' embryos, results presented confirm the real possibility of the future restoration of the colony of pt rabbit, if sufficient number of embryos are cryopreserved.

Turbomolecular Pumped Electron Microscopy of Rubella Virus (In BHK Celis)

1968 ◽  
Vol 26 ◽  
pp. 204-205
Author(s):  
A. B. Taylor ◽  
G. C. Cole ◽  
M. A. Holcomb ◽  
C. A. Baechler
Keyword(s):  
Electron Microscopy ◽  
Rubella Virus ◽  
Phosphate Buffer ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Basal Medium ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Input Multiplicity ◽  
Baby Hamster Kidney Cells

An aliquot from a continuous fermenter culture of baby hamster kidney cells (BHK-21 Clone PD-4) (Wistar) maintained in Ca free Eagle's Basal Medium containing 2% Kaolin adsorbed fetal calf serum was planted in spinner flasks at 300,000 cells per ml, total volume 600 ml. After equilibration for one day at 35°C to insure that cells were in log phase, the culture was infected with the M-33-AGMK25 BHK-219 strain of rubella at an input multiplicity of about 6 TCID50 per cell. The virus was identified with specific rubella antiserum.Preliminary experiments had shown that such cultures would reach a peak or plateau HA titer of approximately 1:64, 24 hrs after inoculation and would continue to yield virus for 6 to 12 days. One hundred ml aliquot harvests were withdrawn daily and the culture was returned to volume with growth medium and incubation continued. The harvested cells were spun down rapidly at 2500 rpm per 15 mins., fixed in 3.7% gluteraldehyde in Ca free phosphate buffer saline, and post fixed in osmium tetraoxide. After dehydration, the cells were embedded in Epon 812 and cured approximately 20 hrs at 60°C.

Fish kidney cell line in response to heat shock

1992 ◽  
Vol 50 (1) ◽  
pp. 704-705
Author(s):  
Li-Chu Tung ◽  
Yung-Reui Chen ◽  
Shiu-Nan Chen ◽  
Guang-Hsiung Kuo
Keyword(s):  
Heat Shock ◽  
Cell Line ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Uranyl Acetate ◽  
Ultrastructural Changes ◽  
Heat Shock Treatment ◽  
Acanthopagrus Schlegeli ◽  
Cacodylate Buffer ◽  
Fish Kidney

In the present study, the ultrastructural changes of BPK cells, a fibroblast-like cell line, derived from the kidney of juvenile black porgy Acanthopagrus schlegeli, under heat shock treatment are described.The BPK cells were maintained in L-15 medium supplemented with 10% fetal calf serum and 0.15 M NaCl at 28|C2. The heating was carried out in precalibrated water baths. Monolayers of cells, grown on coverslips in parafilm-sealed petri dishes were submerged under water for 30 min at 40|C treatments. Cells were fixed in 2.5% glutaraldehyde in 0.1 M cacodylate buffer supplemented with 6.6% sucrose, postfixed in 1% OsO4 and flat embedded in Spurr’s resin. Silver section were cut parallel to the substratum, stained with uranyl acetate and Reynold’s lead citrate, and examined in a Hitachi H-600 electron microscope at 75 KV.

scholarly journals Fetal calf serum contains activities that induce fetal hemoglobin in adult erythroid cell cultures

Blood ◽  
1990 ◽  
Vol 75 (9) ◽  
pp. 1862-1869 ◽  
Author(s):  
P Constantoulakis ◽  
B Nakamoto ◽  
T Papayannopoulou ◽  
G Stamatoyannopoulos
Keyword(s):  
Cell Cultures ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Fetal Hemoglobin ◽  
Erythroid Cell ◽  
Erythroid Progenitors ◽  
Globin Mrna ◽  
Erythroid Progenitor ◽  
Fetal Sheep ◽  
Gamma Globin

Abstract Cultures of peripheral blood or bone marrow erythroid progenitors display stimulated production of fetal hemoglobin. We investigated whether this stimulation is due to factors contained in the sera of the culture medium. Comparisons of gamma/gamma + beta biosynthetic ratios in erythroid colonies grown in fetal calf serum (FCS) or in charcoal treated FCS (C-FCS) showed that FCS-grown cells had significantly higher gamma/gamma + beta ratios. This increase in globin chain biosynthesis was reflected by an increase in relative amounts of steady- state gamma-globin mRNA. In contrast to its effect on adult cells, FCS failed to influence gamma-chain synthesis in fetal burst forming units- erythroid (BFU-E) colonies. There was a high correlation of gamma- globin expression in paired cultures done with C-FCS or fetal sheep serum. Dose-response experiments showed that the induction of gamma- globin expression is dependent on the concentration of FCS. These results indicate that FCS contains an activity that induces gamma- globin expression in adult erythroid progenitor cell cultures.

scholarly journals Fetal calf serum enhances in vitro production of Bos taurus indicus embryos

2011 ◽  
Vol 75 (3) ◽  
pp. 429-433 ◽  
Author(s):  
F.G. Leivas ◽  
D.S. Brum ◽  
S.S. Fialho ◽  
W.P. Saliba ◽  
M.T.T. Alvim ◽  
...  
Keyword(s):  
Fetal Calf Serum ◽  
Bos Taurus ◽  
Calf Serum ◽  
In Vitro Production ◽  
Bos Taurus Indicus ◽  
Vitro Production
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