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Biomedical subjects

R H Glass

Publications and source records attributed to R H Glass.

15 recordsLinked to original sources

Maturation of immature oocytes by coculture with granulosa cells.

To increase the number of embryos available for transfer, immature human oocytes were cocultured with granulosa cells from preovulatory follicles. Greater numbers of immature oocytes incubated with granulosa cells had dispersion of the cumulus and corona cells compared with immature oocytes cultured in media alone. Fifty-four percent of immature oocytes were fertilized after coculture with granulosa cells compared with 20% fertilization of immature oocytes cultured without granulosa cells. There were no cases in which only embryos developed from immature oocytes were transferred, and thus we could not determine if the immature oocytes could contribute to a pregnancy.

Cell Communication

The resazurin reduction test provides an assessment of sperm activity.

OBJECTIVE: The objective of the study was to determine if reduction of the dye resazurin by semen could be correlated with the concentration of motile sperm. DESIGN: After assessment of sperm count and motility, specimens were incubated for 1 hour with resazurin (25 micrograms/mL of semen) and visual color changes indicative of dye reduction noted. SETTING: Specimens were obtained from men seeking care for infertility at one of four sites: (1) University of California, San Francisco (UCSF) In Vitro Fertilization Program; (2) UCSF Andrology Laboratory; (3) a gynecological practice in Maine; and (4) a private andrology laboratory in Southern California. PATIENTS: Individuals were self-selected by their desire to have a semen analysis in conjunction with the diagnosis or treatment of infertility. INTERVENTIONS: None. MAIN OUTCOME MEASURE: The reduction of the dye resazurin and its correlation with motile sperm density. RESULTS: When the motile sperm concentration was greater than or equal to 20 X 10(6)/mL, 86% of specimens produced a positive color change. Conversely, 86% of specimens with a motile sperm concentration of less than 20 X 10(6)/mL either did not change color or changed only over a narrow range. CONCLUSION: Reduction of resazurin offers an assessment of the active sperm in a specimen without the need to do a sperm count or evaluation of motility.

Humans

Spontaneous cure of male infertility.

Four of sixteen couples whose infertility was thought to be due to a male factor achieved a pregnancy without treatment. Pregnancy without therapy, despite compromised sperm counts or motilities, has been documented by a number of other authors. Despite this information many studies concerning the treatment of male infertility neglect to include control groups, and pregnancies which occur are credited solely to the therapy. Such claims of therapeutic success should be viewed with caution.

Adult

Habitual abortion.

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Abortion, Habitual

Intrauterine insemination of isolated motile sperm.

Semen from infertile males was filtered through liquid albumin, and sperm retrieved from the most distal fraction were used for intrauterine insemination. Compared with the ejaculate, the isolated specimens contained fewer sperm but an increased percentage of motile sperm. The fractions were also free of the debris (white cells, agglutinated sperm, immature forms, and particulate matter) commonly seen in the ejaculates. Despite the improvement in motility none of the 19 women who were treated for a total of 67 cycles became pregnant. It is possible that even the best sperm from a poor specimen may have intrinsic defects that interfere with their ability to fertilize.

Female

Sex preselection.

There have been attempts to select the sex of a child prior to conception in both animals and humans. Centrifugation, microelectrophoresis and density gradient sedimentation have been used to separate X and Y sperm in animals. Only the latter technic has produced a change in sex ratio. In the human, timing of coitus or artificial insemination in relation to ovulation has been the popular method for influencing the sex of a child. This review suggests caution in accepting the claims of success for coital timing. Sperm bearing the Y chromosome can now be identified by quinacrine staining. Using this marker, Ericsson showed that when sperm are allowed to swin into columns of liquid albumin a high percentage of Y-bearing sperm are found in the most distal portion of the column. At this time it has not been shown whether the fraction enriched with Y sperm can produce a preponderance of males.

Animals

Localization of prostaglandin on the plasmalemma of rabbit sperm.

Prostaglandin and prostaglandin receptors were localized on unwashed ejaculate rabbit sperm utilizing ferritin conjugated antibody to prostaglandin. Reaction was obtained over the acrosomal portion of the plasma membrane, with a greater aggregation over the apex of the head. The remainder of the head plasmalemma and flagellum showed no deposits of of reaction product. The reaction did not occur if the sperm were washed prior to incubation with the labelled antisera. However enhanced binding of ferritin was demonstrated if washed sperm were pre-incubated in PGE1. No reaction was seen in similar studies with human sperm.

Animals

Interaction of male and female germ cells of two mammalian species in culture.

When viewed by scanning electron microscopy, the heads of mouse spermatozoa are smaller than those of the hamster. The vitelline microvilli of hamster eggs are longer than those of the mouse egg. Both these factors may contribute to the enhanced interaction of mouse spermatozoa and hamster eggs. Treating unfertilized mouse eggs with Newcastle disease virus causes the viteline microvilli to elongate, thus improving the interaction between mouse eggs and hamster spermatozoa.

Animals

Development of the mouse blastocyst after actinomycin D treatment.

Mouse blastocysts were incubated in vitro with actinomycin D in concentrations of 10-1 to 10-5 mug/ml. After transfer to recipient animals, there was a moderate but significant decrease in implantations at 10-minus 1 to 10-minus 3 and 10-minus 5. There was a severe inhibition of subsequent development at concentrations of 10-minus 1 and 10-minus 2. One of 49 fetuses which developed after actinomycin D treatment had a major malformation; there was one major and one minor abnormality among 95 control fetuses. It is suggested that most of the RNA or protein required to effect implantation is synthesized or coded for prior to the blastocyst state. The moderate but significant decrease in implantations found after actinomycin D treatment at this stage may reflect the ability of the drug to reduce cell adhesiveness. This study confirms the impression that the preimplantation embryo is relatively impervious to teratogenic agents, even one as potent as actinomycin D.

Abnormalities, Drug-Induced