Sex preselection through albumin separation of sperm.

OBJECTIVE: To determine if passage of sperm through columns of liquid albumin before their use in artificial insemination could affect the sex ratio at birth. DESIGN: Sperm were isolated by layering over columns of liquid albumin. The isolated fractions were inseminated into the uterus on the presumptive day of ovulation. SETTING: Patients were treated in 65 clinical practices (Sperm Centers) located both in this country (57) and abroad (8). PATIENTS: Individuals were self-selected by their desire to have a child of a specified sex. MAIN OUTCOME MEASURE: The sex of offspring resulting from the insemination of isolated sperm. RESULTS: Insemination with sperm isolated to enhance male sex preselection produced 71% and 76% males depending on the technique. Use of isolated sperm in women who were taking clomiphene citrate was associated with a 69% birth of females. CONCLUSIONS: Separation of sperm on columns of liquid albumin can affect the sex ratio at birth.

PIP: Fertility specialists examined data on 1407 births resulting from artificial inseminations with sperm separated on albumin columns and occurring at 59 sperm centers in the US and at 8 sperm centers in other countries to determine whether albumin separation can effectively affect the sex ratio. The patients had already had at least 2 children of the same sex and wanted a child of the opposite sex and consciously opted to use sex preselection techniques. Male newborns predominated (72% vs. 28%). The modified protocol 3 resulted in a higher percentage of male births than did protocol 3 (76% vs. 71%), but the difference was insignificant. Both protocols significantly influenced the sex ratio (71% vs. 29% for protocol 3, and 76% vs. 24% for modified protocol 3; p .0001). For female sex preselection, protocol 2 included administration of clomiphene citrate to the woman to induce ovulation and less complete sperm separation. It resulted in a significantly higher percentage of female births than male births (69% vs. 31%; p .0003). 5.7% of a representative sample (sperm centers in Dallas, Texas, and Lathrup Village, Michigan) were lost to follow-up, but these 547 couples were apparently the same as the entire sample. These findings indicated that the albumin separation method does influence the sex ratio at birth, but its use is likely limited since it is not 100% effective.

Sex ratio of male sex preselected children born to couples with exclusively female offspring.

The present study was undertaken to determine if couples with exclusively female children had a decreased probability of a male child after using male sex preselection. Selection criteria for subjects necessitated that couples have had only female children previously and had produced a child at one of 14 centers after using protocol 3 (n = 70) and modified 3 (n = 28) male sex preselection. Prior to sex selection, protocol 3 couples produced a combined total of 135 female children for an average of 2.01 (range 1-4) females per couple; modified 3 couples produced a combined total of 62 female children for an average of 2.21 (range 1-4) females per couple. The normal approximation to the binomial distribution was used to determine significant differences between the sex ratios prior to and after male sex preselection. For couples using protocol 3 there were significant differences in the sex ratio prior to sex preselection (0%) and after sex preselection (73.0%) (p less than .00003). There were also significant differences between the sex ratio prior to modified 3 (0%) and after sex preselection (86%) (p less than .00003). Couples using male sex preselection do not have a decreased probability of a male child if they have had exclusively female children.

Couples with exclusively female offspring have an increased probability of a male child after using male sex preselection.

The present study was undertaken to determine whether couples with one or more daughters and no sons had an increased probability of having a male child after using male sex preselection. The sex ratio of children born to couples after using 'protocol 3' (n = 70) or 'modified protocol 3' (n = 28) male sex preselection at one of 14 centres was determined. The normal approximation to the binomial distribution was used to determine significant differences between these sex ratios and the established sex ratio for children born to couples with one previous daughter and no sons. The sex ratios of both protocol 3 sex-preselected children (73.0%; P less than 0.0001) and modified protocol 3 sex-preselected children (86%; P less than 0.0001) were significantly different to the established sex ratio (control) for a current child born to parents with one previous daughter and no sons (50.1%). Couples with one or more daughters and no sons will have an increased probability of a male child after using male sex preselection.

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.

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.

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.

PIP: An albumin column technique for isolating motile sperm from ejaculate was used to artificially inseminate 19 women, members of couples who suffered long-term infertility (4.5 years, range 1-10 years) thought to have a male factor as the primary cause. Semen from infertile males was filtered through liquid albumin, and sperm from the most distal end of the fraction were used for intrauterine insemination. Th isolated specimens contained fewer sperm but an increased percentage of motile sperm compared with the ejaculate. Women were treated for a total of 67 cycles (8 for only 1 or 2 cycles; none for more than 8 months). Though the sperm fraction isolated on albumin columns showed good motility, numbers apparently adequate for conception (as gleaned from previous animal and human artificial insemination studies), more uniform morphology, and absence of seminal debris, no pregnancies occurred during treatment cycles. Possibly even the best sperm from a poor specimen have intrinsic defects that interfere with their ability to fertilize, since previous reports of normal pregnancies following intrauterine insemination have been documented.

Isolation and storage of progressively motile human sperm.

Human sperm progressively motile and free of seminal debris can routinely be isolated, during a 1.5 hour process, from semen of normal and subfertile men. Serum albumin, in a vertical column, acts to filter sperm based on their swimming abilities of forward progression. The isolated sperm constitute a population that can be inseminated as an aid to male subfertility or, to enhance the chances of sperm survival upon deep-freeze preservation. The latter use resulted in twice the survival rate, based on sperm motility, or random populations of control sperm.

[X and Y spermatozoa].

PIP: In humans, the X- or Y-bearing spermatozoa determine the sex of the offspring. The Y-bearing sperm contain 4% less DNA, 7% less surface area, swim faster, and can be distinguished by their fluorescence in certain dyes from the X-bearing sperm. It is possible to enrich semen in Y-bearing sperm by lowering the temperature, or depositing them in a column containing different concentrations of albumin. Some other methods that have been tried without success include pH, electric charge, centrifugation, sedimentation, flotation, gel filtration, immunological methods, and enzyme tests, in human, rabbit, bull, and mouse semen. It is believed that the only workable way to accumulate data in humans is to try sex selection techniques on couples consulting for infertility. These techniques usually increase the proportion of healthy, mobile sperm; it is unknown whether the natural selection of the cervical mucus barrier performs the same function. It is possible now to determine the sex of the fetus accurately from an amniocentesis specimen, and abort the unwanted sex, but this practice is not being done. Factors known to alter the sex ratio include diet, vaginal pH, father's profession, time of the year, annual rainfall, time of the menstrual cycle, and birth rank. Probably if the precise sex, timing, and number of children desired could be obtained, fewer children would be born.^ieng

Fertilization of rabbit ova in vitro by sperm with adsorbed Sendai virus.

Fertilization occurs when rabbit ova are cultured in vitro with epididymal sperm to which Sendai virus is adsorbed. These sperm do not require capacitation in vivo in order to fertilize. Evidence for fertilization is penetration of sperm, the appearance of two polar bodies and pronuclei, and cleavage through the eight-blastomere stage. The viruses attach almost exclusively to the sperm acrosome, with resultant head-to-head agglutination of the sperm.