Biomedical subjects

G F Amborski

Publications and source records attributed to G F Amborski.

At least 19 recordsLinked to original sources

PubMed1991

Effects of stage of the bovine oestrous cycle on in-vitro characteristics of uterine and oviductal epithelial cells.

The objective of this experiment was to evaluate the effects of stage of the bovine oestrous cycle on in-vitro morphology, growth and monolayer foundation of uterine and oviductal epithelial cells. Epithelial cells were isolated from the uterus and oviducts collected from cyclic cattle on the day of oestrus (Treatment A), and between days 4 to 6 (Treatment B), days 8 to 10 (Treatment C) and days 14 to 16 (Treatment D) of the oestrous cycle. The morphological development, per cent cell viability and cell attachment were evaluated during primary culture and after the first and third subpassages. The highest per cent cell viability and cell attachment during primary culture, respectively, were noted in Treatment B for both uterine (87.7 and 87.5%) and oviductal (88.4 and 87.2%) cell populations. Uterine epithelial cell populations in Treatments C and D, respectively, had the lowest viability (76.5 and 68.8%) and attachment (10.8 and 10.5%) during primary culture. There were marked improvements in cell viability and cell attachment following the first subpassage (P less than 0.001) compared with primary cultures for both uterine and oviductal cells. These results indicate that the stage of the oestrous cycle has dramatic effects on uterine and oviductal epithelial cell morphology and developmental patterns during primary in-vitro cultures. The stage of the oestrous cycle when cells are collected may be more important than was once realized when culturing early stage embryos in vitro.

Animals↗

PubMed1989

Serological detection of multiple retroviral infections in cattle: bovine leukemia virus, bovine syncytial virus and bovine visna virus.

Individual experimental animals used in our studies on bovine leukemia virus (BLV) are routinely screened for the presence of antibodies to the three bovine lymphotropic retroviruses. We utilized these screening methods to examine frozen sera from eight herds for antibodies to BLV, bovine visna virus (BVV) and bovine syncytial virus (BSV). Serum samples from 235 animals in four dairy and four beef herds were analyzed. Detection methods used included indirect fluorescent antibody tests of virus-infected cell cultures (BLV, BSV, BVV) and agar gel immunodiffusion (BLV). Sera from the BLV-infected animals in the dairy herds showed the highest single (50%, 49/97) and multiple (30%, 29/97) infections compared with 5% (7/138) and less than 1% (1/138), respectively in the beef herds. Single BVV infections were not detected in the dairy herds, but 11% (11/97) of the sera contained antibodies to BVV plus BLV or BSV. Five sera from beef cattle had antibodies only to BVV and four were obtained from one herd. Only one beef serum of the 138 tested demonstrated multiple antibodies (BLV, BVV).

Animals↗

PubMed1989

In-vitro development and implantation of human embryos following culture on fetal bovine uterine fibroblast cells.

Human zygotes resulting from IVF were placed in two different culture systems to evaluate in-vitro development and to establish pregnancies in patients following embryo replacement. Treatment A (control) consisted of culturing zygotes in a modified Earle's Balanced Salt solution while treatment B consisted of culturing zygotes on a monolayer of fetal bovine uterine fibroblasts in this same culture medium. At the time of embryo replacement, embryos within treatments A and B had 3.7 and 4.3 blastomeres present, respectively. After 24 h in culture, the cellular fragmentation rate for treatment A embryos was 0.85 which was greater (P less than 0.05) than the fragmentation rate of 0.40 for embryos within treatment B. The incidence of implantation for patients whose embryos were given treatment A was 17.0% which was lower (P less than 0.05) than 35% for those given treatment B. Implantation rates increased with time in culture (43%) for treatment B embryos. Culture by treatment B of three-pronucleate zygotes resulted in 7/9 and 4/9 reaching the blastocyst and expanded blastocyst stages, respectively, whereas only 1/26 three-pronucleate zygotes cultured using treatment A reached either of these stages.

Blastocyst↗

PubMed1988

Molecular studies of T-lymphocytes from cattle infected with bovine leukemia virus.

Although bovine leukemia virus (BLV) is mainly associated with infections of B-lymphocytes, we have previously reported the statistically significant increase in the T-lymphocytes obtained from BLV-infected asymptomatic aleukemic (AL) cattle. In this report the presence of BLV provirus in the DNA of immunoaffinity purified T-lymphocytes from AL animals was assessed using a highly specific radiolabelled (32P) BLV-DNA provirus probe and solid phase DNA hybridization. The BLV provirus was found in the DNA of the peripheral blood mononuclear cells of all AL animals tested and three of the four purified T-lymphocyte preparations from these animals. The purified T-lymphocyte preparations used in this study contained less than 4% detectable B-lymphocytes. One animal had no detectable B-lymphocytes in the purified T-lymphocyte preparation and the DNA from these cells also gave positive hybridization results. The lymphocyte blastogenesis assay was then used as an indicator of the functional ability of lymphocytes from these BLV-infected AL cattle to respond to mitogenic stimuli. The responsiveness of lymphocytes from these animals to the mitogens concanavalin A (Con A), phytohemagglutinin (PHA), and pokeweek mitogen (PWM) was comparable to that of lymphocytes from BLV-negative animals when changes in 3H-thymidine uptake (c.p.m.) were used as measurement of mitogenic-induced blastogenesis. This indicated that infection of the T-lymphocytes by BLV does not appear to alter the overall response of the lymphocyte populations to mitogenic stimuli. High levels of spontaneous blastogenesis in the absence of mitogenic stimulation were observed for lymphocyte preparations of AL animals. The reason for this proliferation of lymphocytes is unclear; however, sera from these AL animals were found to contain a blastogenesis-augmenting factor(s) when added to lymphocytes from BLV-negative control animals in the presence of Con A, PHA and PWM.

Animals↗

PubMed1988

Enumeration of T and B lymphocytes in bovine leukemia virus-infected cattle, using monoclonal antibodies.

Monoclonal antibodies and microfluorimetry were used to determine the absolute number of B and T lymphocytes in the blood of bovine leukemia virus (BLV)-infected cows. The blood lymphocyte populations from BLV-infected cows were significantly higher than those from BLV-negative cows. The increase in the lymphocyte population in 3 BLV-infected nonlymphocytotic cows was attributed to a significant increase in the number of T lymphocytes; in 3 BLV-infected persistently lymphocytotic cows, the increase was attributed to a significant increase in the number of B and T lymphocytes. One persistently lymphocytotic cow had a high lymphocyte count, and lymphocytes from this cow contained cells that appeared to stain with markers specific for bovine B and T lymphocytes. We concluded that infection of cattle with the B-cell lymphotropic retrovirus, BLV, not only affected B cells, but also T cells.

Animals↗

PubMed1987

Isolation of a retrovirus from the American bison and its relation to bovine retroviruses.

Tissue samples were removed at necropsy from five American bison (Bison bison) with clinical signs of a disease resembling malignant catarrhal fever (MCF). Using cell-associated virus techniques, attempts were made to isolate viruses from these tissues by culturing them directly or by co-culture with bovine fetal cells. Among the viruses isolated was one which was syncytiogenic and multiplied in bovine fetal spleen cells and remained highly cell-associated. The presence of reverse transcriptase activity indicated that it was a retrovirus. Also, it had antigenic cross activity with bovine syncytial virus, but not with bovine leukemia or bovine maedi-like retroviruses. We do not attribute a direct causative role of this retrovirus to MCF, but indirect relationships are possible.

Animals↗

PubMed1986

Enrichment of T lymphocytes from bovine peripheral blood mononuclear cells using an immuno-affinity depletion technique ("panning").

A simple and efficient method to enrich bovine T lymphocytes from peripheral blood mononuclear cells (PBMC) by immuno-affinity depletion ("panning") has been developed. The PBMC were initially separated by density gradient centrifugation on Histopaque of density 1.077 g/ml. The T lymphocyte subset was then separated from PBMC by depletion of membrane immunoglobulin (Ig) bearing cells which had an affinity for anti-Ig antibodies bound to polystyrene tissue culture flasks. An average of 95% of the nonadherent "panned" cells were identified as T lymphocytes using a label of peanut agglutinin conjugated with fluorescein isothiocyanate (PNA-FITC). Two percent of the PNA negative cells were Ig bearing cells. The average yield was 50% of the original T lymphocytes found in the PBMC population, and the cell viability as assessed by trypan blue exclusion was greater than 95%. The separation took approximately 2 hours, and the total number of T lymphocytes recovered from 40 ml of blood was in the range of 20-40 X 10(6).

Animals↗

PubMed1984

In vitro prostaglandin E2 stimulation of 45Ca mobilization from chick bone.

The ability of prostaglandin E2 (PGE2) to mobilize 45Ca from chick embryo long bones was assayed in an in vitro bone culture system. Concentrations of PGE2 tested ranged from 10(-9) to 10(-5) M. The PGE2 was effective in stimulating release of 45Ca from prelabelled bones at all concentrations tested except at 10(-9) M. In addition, stimulation of 45Ca release could be produced daily for 4 consecutive days of PGE2 culture-pulsing at what appeared to be the optimal PGE2 concentration, 10(-7) M. We conclude, as in mammals, PGE2 is a potent stimulator of calcium mobilization from avian bone. The potential involvement of prostaglandins in eggshell formation is discussed.

Animals↗

PubMed1984

Prevalence of bovine herpesvirus-1, bovine viral diarrhea, parainfluenza-3, goat respiratory syncytial, bovine leukemia, and bluetongue viral antibodies in sheep.

Sera from healthy sheep were collected in January and March 1982 from flocks of sheep located in southwestern and southeastern Louisiana. These sera were tested for bovine herpesvirus-1 (BHV-1), bovine viral diarrhea virus (BVDV), parainfluenza-3 (PI-3) virus, and goat respiratory syncytial virus (GRSV) antibodies by microtitration virus-neutralization test. The sera were tested also for bovine leukemia virus (BLV) and bluetongue virus (BTV) antibodies by immunodiffusion tests. The number of flocks with seropositive sheep for each virus were: 2/8 (25%) for BVDV; 8/8 (100%) for PI-3 virus; 7/8 (87.5%) for GRSV; and 6/8 (75%) for BTV. Seropositive rates for each virus for the individual sheep tested were: 4/158 (2.5%) for BVDV; 117/158 (74.1%) for PI-3 virus; 77/158 (48.7%) for GRSV; and 21/158 (13.3%) for BTV. All sheep were seronegative for BHV-1 and BLV.

Animals↗

PubMed1979

Equine infectious anemia virus: development of a simple reproducible method for titrating infectivity of the cell-adapted strain.

An equine dermal cell line between the 14th and 30th subpassages was used to develop a reproducible method of titrating the infectivity of the cell-adapted strain of equine infectious anemia virus (EIAV). Cells inoculated with EIAV were subcultured or fed once each week and were monitored for the production of P26 antigen of EIAV in supernatant fluids. Ultrathin sections were prepared once each week and were examined for the detection of budding virus-like particles (VLP). The VLP could not be detected in the infected cells subcultured once each week. When the medium was changed once week and when the cells not transferred, budding VLP were detected routinely after 3 to 4 weeks, Feeding of the infected cells once each week was used to establish the infectivity assay. The reproducibility of the assay on a frozen viral stock preparation was within an endpoint of one tenfold dilution.

Animals↗

PubMed1974