PubMed Health⌕ Search

Biomedical subjects

T B Pool

Publications and source records attributed to T B Pool.

At least 19 recordsLinked to original sources
PubMed1994

High continuing pregnancy rates after in vitro fertilization-embryo transfer using medium supplemented with a plasma protein fraction containing alpha- and beta-globulins.

OBJECTIVE: To evaluate both the laboratory and clinical outcomes after IVF-ET using culture media supplemented with a plasma protein fraction (PPF, Plasmatein; Alpha Therapeutics, Los Angeles, CA) containing albumin and significant amounts of alpha- and beta-globulins. DESIGN: One-year clinical trial of a PPF with high globulin content as a medium supplement during IVF, embryo growth, and ET. SETTING: Fertility Center of San Antonio, a private, office-based center for assisted reproduction. PATIENTS: Ninety-eight couples, with women ranging in age from 26 to 46 years, undergoing 103 ovum retrievals for IVF-ET as treatment for infertility because of tubal factor, endometriosis, anovulation, uterine or cervical factor, male factor, and unexplained causes. MAIN OUTCOME MEASURES: Fertilization rate, zygote cleavage rate, clinical pregnancy rate (PR), continuing PRs, and implantation rates. RESULTS: Supplementation with PPF in insemination, growth and transfer medium resulted in a clinical PR of 41.5% per transfer with continuing PRs of 35.2% per retrieval, 37.2% per patient, and 38.7% per transfer. CONCLUSIONS: A PPF containing significant amounts of alpha- and beta-globulins can serve as an effective protein supplement to IVF medium, with outcomes manifested as high continuing PRs. These data indicate a potential role for glycoprotein components of serum in supporting healthy embryo growth in vitro, although the mechanism may relate more to the general physicochemical properties of this fraction than to the actions of a specific component.

Adult↗
PubMed1990

Zygote intrafallopian transfer with "donor rescue": a new option for severe male factor infertility.

Clinical pregnancies have been initiated by ZIFT using zygotes produced by reinsemination of oocytes with donor sperm ("donor rescue") after an initial 15- to 20-hour exposure to husband's sperm. A total of 54 oocytes from four couples experiencing failed fertilization by husband's sperm were reinseminated with donor sperm, resulting in 38 zygotes (70.4% fertilization). Four zygotes were transferred during ZIFT in each case and resulted in two (50%) continuing pregnancies. Additional zygotes from donor reinsemination were cryopreserved for each couple. Donor rescue expands the utility of ZIFT as a treatment for male factor infertility.

Embryo Transfer↗
PubMed1990

Zygote intrafallopian transfer as a treatment for nontubal infertility: a 2-year study.

Zygote intrafallopian transfer (ZIFT) was used as a treatment for long-standing nontubal infertility for a 2-year period. The overall clinical pregnancy rate for 114 tubal transfers was 40.4% with a delivery/ongoing rate of 34.2%. Concurrent use of in vitro fertilization and embryo transfer (IVF-ET) for tubal factor infertility gave significantly lower clinical pregnancy and delivery/ongoing rates (21.1% and 15.8%, respectively). The use of gamete intrafallopian transfer (GIFT) for nontubal infertility yielded a 32% clinical pregnancy rate and a 26% delivery rate for 53 transfers. Zygote intrafallopian transfer resulted in an implantation rate per zygote of 17% overall compared with 8.1% per embryo for IVF-ET and 11.2% per oocyte for GIFT. The transfer of three zygotes per patient gave the same clinical pregnancy rate as the transfer of four while reducing the incidence of multiple gestation from 19% to 7.8% per transfer. No significant decline in the clinical pregnancy or delivery rate was seen with ZIFT in women aged 25 through 39.

Adult↗
PubMed1988

Impaired proliferation response after PDGF induction in fibroblasts from Hutchinson-Guilford Progeria syndrome.

Fibroblasts from a Hutchinson-Guilford Progeria Syndrome (HGPS) patient were compared to normal human fibroblasts to determine if differences existed in growth factor mediated cell proliferation. Cultures of progeric fibroblasts were exposed individually to platelet-derived growth factor (PDGF), epidermal growth factor (EGF), platelet poor plasma (PPP) and fetal bovine serum (FBS). Autoradiographic studies using 3H thymidine showed that progeric fibroblasts had similar labeling indices relative to controls after exposure to FBS and EGF. In contrast, progeric cells made competent with PDGF and later treated with 5% PPP had a significantly lower labeling index. This and preliminary observations on fos RNA accumulation suggests the possible existence of a genetic defect in HGPS fibroblasts.

Autoradiography↗
PubMed1986

Embryo cryopreservation in cynomolgus monkeys.

An improved knowledge of cryopreservation of primate embryos will have important research and clinical application. Fifty-six 4- to 8-cell in vitro fertilized embryos were frozen in HEPES-buffered Tyrode's solution containing 1.5 M dimethylsulfoxide (DMSO) and cooled at the rate of 0.3 degrees C/minute to -39 degrees C before being transferred into liquid nitrogen. Embryos were rapidly thawed at room temperature for 2 minutes. DMSO was diluted with medium in three steps at 5-minute intervals. Of the 56 embryos, 39 (70%) were classified as viable on the basis of surviving the freezing process with greater than 50% of their blastomeres intact. Twelve of the 39 embryos were cultured overnight, and 11 cleaved at least once. Twenty-five embryos were transferred to nine synchronized, unstimulated recipient monkeys 24 to 48 hours after ovulation. Three pregnancies (33.3%) resulted from the nine transfers.

Animals↗
PubMed1985

Vasoactive intestinal peptide increases prolactin messenger ribonucleic acid content in GH3 cells.

The influence of vasoactive intestinal peptide (VIP), TRH, and dexamethasone (Dex) on PRL mRNA was investigated in PRL-producing GH3 cells using cytoplasmic dot hybridization and RNA blot analysis. Total cytoplasmic RNA was transferred to nitrocellulose filter paper and quantitated by hybridization to PRL recombinant DNA probes labeled with 32P. Incubation of GH3 cells with VIP for 25 h increased the content of cytoplasmic PRL mRNA. This increase was dose dependent, being significant at 2 X 10(-8) M and reaching a maximum at 2 X 10(-7) M. VIP at 2 X 10(-9) M had no effect on cytoplasmic PRL mRNA content. TRH (2 X 10(-7) M) also increased whereas Dex (2 X 10(-7) M) decreased the content of PRL mRNA. The inhibitory effect of Dex (2 X 10(-7) M) on cytoplasmic PRL mRNA was reversed by VIP (2 X 10(-7) M). Changes in medium PRL levels after these various hormone treatments paralleled those changes observed in PRL mRNA content. Examination of total poly(A)+ RNA demonstrated that incubation with VIP (2 X 10(-7) M) for 6 h increased the content of the mature PRL mRNA and its processing intermediates. Dex (2 X 10(-7) M) decreased the content of all species of PRL mRNA. These data suggest that VIP-stimulated PRL release is the result of an increase in the content of PRL mRNA and its precursors.

Animals↗
PubMed1984

Successful in vitro fertilization and embryo transfer in cynomolgus monkeys.

We have started an in vitro fertilization program in cynomolgus monkeys in an effort to develop an appropriate animal model to improve our knowledge of early embryonic development. In 16 of 25 animals treated with menopausal gonadotropins, preovulatory follicles developed. Follicular aspiration was performed at laparotomy after human chorionic gonadotropin injection. A total of 299 follicles were aspirated, and 251 oocytes were recovered. Oocytes were cultured in 1 ml of growth medium or 100 microliter droplets of medium under mineral oil. Semen samples were obtained by electroejaculation, and the oocytes were inseminated 4 to 24 hours after aspiration. Culture under mineral oil significantly increased the fertilization and cleavage rates. Of 68 embryos produced, 24 have been used in 10 embryo transfers, resulting in two pregnancies.

Animals↗
PubMed1983

Effects of amiloride on tumor growth and intracellular element content of tumor cells in vivo.

The effects of amiloride, a reported inhibitor of serum-stimulated sodium influx, were tested on tumor growth, tumor cell proliferation, and intracellular element content of cancer cells in vivo. We have shown previously that cancer cells have high intranuclear levels of sodium compared to those of their normal counterpart cells and have postulated that such a high level of sodium may be involved in the cancer state. We now report that amiloride, when given in a series of injections, inhibited both H6 hepatoma and DMA/J mammary adenocarcinoma growth in vivo in a dose-dependent fashion and that 3 injections of amiloride at a dose of 1.0 microgram/g body weight into mice bearing H6 hepatomas resulted in a significant decrease in the intranuclear content of sodium but not the content of magnesium, phosphorus, sulfur, chlorine, or potassium as measured by electron probe X-ray microanalysis in the H6 hepatoma cells. Amiloride at dosages as low as 1.0 microgram/g body weight per injection also inhibited tumor cell proliferation as measured by the tritated thymidine autoradiography labeling index. Amiloride caused no changes in the mean profile diameters of metaphase or interphase H6 hepatoma or DMA/J mammary adenocarcinoma cells, suggesting that the action of amiloride on tumor growth was not due to cell volume changes. These data show that amiloride both inhibited tumor growth and decreased the proliferation of the tumor cells in the H6 hepatomas which was correlated with a decreased intranuclear sodium content.

Adenocarcinoma↗
PubMed1982

Quantitative morphological analysis of proliferating and nonproliferating subpopulations of IMR-90 fibroblasts during aging in vitro.

Early-, mid- and late-passage cultures (population doubling levels 12, 35, and 51, respectively) of IMR-90 fibroblasts were exposed to 3H-thymidine for 48 h prior to fixation in situ for morphometric analysis in order to determine quantitatively what ultrastructural changes accompany the loss of proliferative capacity during aging in vitro. Analysis of autoradiographs, both at the light and electron microscopic levels, with an image analyzer followed by ANOVA statistical scrutiny demonstrated that a significant increase in relative cell area, an indicator of cell size, was characteristic of cells unable to incorporate 3H-TdR at both mid- and late-passage, but not at early-passage levels. Nuclear size also increased significantly with progressive passage level but was not related to proliferative capacity. No significant difference in the area fraction of nucleoli per unit area of nucleus or of mitochondria, Golgi, or lysosomes was seen in either subpopulation at any passage level. Dilated cisternae of rough endoplasmic reticulum in early-passage cells were seen if cells were harvested with trypsin and fixed either before or after centrifugation, but were not seen in labeled or unlabeled cells from any passage level when cultures were fixed in situ. We conclude that a significant increase in cell size is the only significant morphological change associated with the loss of proliferative capacity of IRM-90 fibroblasts. Furthermore, our data indicate that there is no accumulation of secondary lysosomes in human diploid fibroblasts during aging in vitro; we therefore cannot support any hypothesis of aging or proliferative decline that is based mechanistically upon this phenomenon.

Cell Division↗
PubMed1981

Changes in nuclear shape and mitochondrial structure do not accompany the loss of division potential in human fibroblasts in vitro.

Previous studies on ultrastructural changes that occur in cultured human fibroblasts during their in vitro life-span indicate that "senescent" cells characteristically possess structurally altered mitochondria, highly lobed nuclei, and an abundance of secondary lysosomes when compared to early passage cells. In the present study, we demonstrate that improper preparative methods can induce altered mitochondrial morphology in preparations of both IMR-90 and HF730A fibroblasts, regardless of passage level. We also show that nuclei of both living and fixed IMR-90 fibroblasts are ovoid in shape, not lobulate, in well-spread cells, regardless of either the passage level or the proliferative capacity of the cell. Fibroblasts contain lobulated nuclei only when they have not spread completely on the culture substrate. Lobulations can be induced at any passage level by collagenase/trypsin or trypsin/EDTA treatment prior to fixation, but not by cytochalasin B treatment or by cold temperatures. We conclude that any treatment that affects cytoskeleton-membrane-culture substrate interactions will induce this aberrant nuclear morphology, but that this is not indicative of "senescence" and does not relate to proliferative decline.

Cell Cycle↗
PubMed1981

Elemental content in the nucleus and in yolk and yolk-free cytoplasm of developing Rana pipiens oocytes: an x-ray microanalysis study.

Energy dispersive x-ray microanalysis measured the Na, K, Cl, P, Mg, S, and Ca contents (mM/kg dry weight) in the nucleus, yolk-free cytoplasm, and yolk platelets of Rana pipiens oocytes quick frozen in the ovary. The data revealed that significant content changes occur in frog oocytes during intraovarian growth. All elements but Ca changed in content in the nucleus and cytoplasm, while in the yolk platelet only Na content did not change. A nucleus to cytoplasm K gradient develops and increases in magnitude during oocyte growth. The data from this and previous reports lead to the hypothesis that intra-oocytic water and elements undergo changes in state during oocyte growth and that three subcellular Na compartments exist.

Animals↗
PubMed1980

Intracellular concentration of potassium and other elements in vaginal epithelial cells stimulated by estradiol administration.

Electron probe x-ray microanalysis was used to measure cytoplasmic elemental content (in mmoles/kg dry weight) of the basal layer of cells of the vaginal epithelium of ovariectomized rats. Measurements were made both before estradiol injection and at 2 hr, 17 hr, and 24 hr after estradiol administration. Mitotic figures first appeared in the basal cell layer at 24 hr. During the course of the study significant time-dependent differences were seen in the content of all elements mesured. A pattern of change in cytoplasmic content was seen for Na, P, S, and Cl; all of which decrease significantly by 17 hr and then return to approximately the nonstimulated concentration by 24 hr. On the other hand K, and to a lesser extent Mg, show an early and continued increase in cytoplasmic content after estadiol injection. Thus, the marked increase in the intracytoplasmic content of K in the estradiol treated cells suggests that K, or the ratio of Na to K, may be directly or indirectly involved in growth stimulation.

Animals↗
PubMed1980

Changes in proliferation and surface morphology in the rat ileum in response to total parenteral nutrition.

To find out how the ileum adapts to total parenteral feeding, two experiments were performed. In the first experiment rats were given total intravenous feeding for 10 days. The animals were injected with tritiated thymidine (1 muCi/g body weight) 1 hour before being killed. Portions of the ileum were used for (1) radioautography, (2) analysis of the tissue DNA content, (3) specific activity of the DNA, and (4) scanning electron microscopy. The DNA content of ileum was decreased 72% while the specific activity of DNA was increased 289% in the i.v. fed rats. In the second experiment rats were given total intravenous feeding for 10 days. The animal were injected with colchicine (1 mg/kg body weight) 3 hours before being killed. The number of labelled cell nuclei per ileal crypt section was significantly decreased by parenteral feeding as was the number of colchicine collected metaphase figures. Light microscopy revealed the crypt and the villus height to be shorter and the number of goblet cells per unit surface area to be increased in parenterally fed rats as compared to those fed solid food orally. Enterocytes of the exfoliative zone from the ileal villi of rats fed solid food showed three distinct types of surface architecture whereas those from i.v. fed rats all possessed abundant microvilli. No bacteria were seen in ilea of i.v. fed animals but many were seen embedded in enterocytes from orally fed rats. Because the amount of DNA per cell is known to be constant, we concluded that the overall number of cells in the lieum decreased about 72% in the i.v. fed rats and that cell proliferation in the crypts, although significantly deceased, was still supporting an epithelial renewal process.

Animals↗
PubMed1980

Evaluation of a preparative method for x-ray microanalysis of soft tissues.

We have adapted the cryopreparative methods designed for the radioautography of diffusible substances (Stumpf and Roth, 1966, 1967) to produce freeze-fried sections of soft tissues for electron probe microanalysis. This report concerns the evaluation of these methods for use with X-ray microanalysis with particular reference to: (1) the preservation of cellular morphology, (2) the introduction of structural artefact by ice crystal formation, (3) the preservation of natural elemental distributions under experimental conditions where known quantities of diffusible elements are present, (4) the effects of ice crystal formation upon possible artefactual elemental redistributions, (5) the effect of section thickness on elemental quantitation, and (6) the effect of tissue excision on element translocation. Freeze-dried sections were prepared from mouse pancreas and from 20% (w/v) solutions of bovine serum albumin (BSA) and gelatin containing known amounts of inorganic salts and were analysed in a scanning electron microscope fitted with energy dispersive X-ray detecting equipment. With the preparative methods used, the morphology of pancreatic acinar cells was well preserved. Acinar cell boundaries, nuclear boundaries, chromatin, nucleoli, ergastoplasm and zymogen granules were readily discernible. Ice crystals were present within sections of BSA, but 70% had a cut surface area < 1 micrometer2 and over 90% had a cut surface area of < 2 micrometer2. Characteristic peak-to-continuum ratios of elements in the BSA sections remained constant over an order of magnitude change in magnification. Elemental redistributions were not detected until the magnification was such that the analysed area fell totally within the confines of a single ice crystal. No differences in elemental peak-to-continuum values were obtained between 2 and 4 micrometer sections of the gelatin-salt solution. Tissue excision did not cause element translocations when compared to tissues frozen in situ. We conclude that this method is valid for preparing tissues for microanalysis under our conditions (analysis of nuclear, cytoplasmic, and secretory compartments) and is limiting only when analyses are conducted at very high magnifications.

Animals↗