Pituitary response to LHRH in hypothyroid women.

This study was designed to characterize pituitary function in premenopausal women during hypothyroid and euthyroid periods. Six subjects with basal thyroid-stimulating hormone (TSH) levels above 10 microU/ml were studied. Estradiol (E2), prolactin (PRL), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) levels were measured by radioimmunoassay at 30, 15, and 0 minutes before infusion. Pituitary function was evaluated by rapid infusion of 10 micrograms of luteinizing hormone-releasing hormone (LHRH) every 2 hours for a total of 6 hours. Samples were then obtained for FSH and LH every 30 minutes for the duration of the 6-hour study. A significant elevation in basal gonadotropins was observed during the hypothyroid period regardless of basal E2 levels in all 6 subjects (P < .01). Basal PRL levels were not significahtly different during the 2 periods (P > .05). In all 6 patients pituitary sensitivity and reserve correlated significantly with basal E2 levels (P < .05), but were not altered by the hypothyroid condition. It is concluded that pituitary responsiveness to LHRH is not altered in hypothyroid women, but that chronic elevation of basal gonadotropins may in part explain the anovulation that so often accompanies this disorder.

Gonadotropin secretion and hypothyroidism.

Hypothyroidism is generally associated with hypogonadotropism. Occasionally, however, increased gonadotropin concentrations are encountered. The mechanisms, presumably hypothalamic, which determine the gonadotropin shift are unclear. A case report of hyperprolactinemic hypothyroidism with associated hypergonadotropism is presented. The previous literature is reviewed. It appears that hypothyroidism is generally associated with a decrease in the gonadogropin secretion. However, hypothyroidism with exxagerated hTSH secretin and hyperprolactinemia can be associated with increased gonadotropin secretion.

Improved glucose control in nonhospitalized pregnant diabetic patients.

Methods for management of diabetic pregnancy in the outpatient setting require strict glucose control. To assess the effect of diet and injection of short and intermediate acting insulin on glucose, diabetic patients tested their urine daily for glucose and had biweekly serum glucose tests. A brief metabolic ward study in 9 diabetic patients during the third trimester yielded hourly glucose determinations. These results defined the range of serum glucose over a 24-hour period. Glucose data on 6 normal third trimester women also came from hourly glucose values. Glucose results of normal and diabetic subjects were similar. A 16th subject with diabetic eye, renal, and foot complications is included as a case report to illustrate management technics. Infants of the diabetic women had no perinatal mortality, morbidity, or macrosomia and thus differ from an earlier study where glucose was not strictly controlled. The data suggest hospitalization can be short and low perinatal morbidity and mortality are possible with this outpatient method of management of the pregnant diabetic patient.

Sperm hyperactivation as quality control for sperm penetration assay.

The sperm penetration assay (SPA) is subject to considerable variation, and controls are needed to verify the accuracy of the results. It is proposed that sperm hyperactivation (HA) can serve as a quality control check for the SPA. The objective was to determine if there was an association between the SPA outcome and sperm HA measured at various times during the SPA procedure. The data showed a significant correlation between percent sperm HA and percent zona-free oocyte penetration by sperm preincubated for three hours prior to sperm-oocyte interaction (short preincubation). Some sperm hyperactivity was observed in liquefied raw semen samples, but this was insignificantly related to SPA results. Low correlation was observed between SPA results and sperm HA determined immediately after centrifuge washing of sperm. The results suggest that it is possible to utilize sperm HA measured immediately after the sperm-oocyte interaction period as a quality control check of SPA results.

Spontaneous pregnancy during the infertility evaluation.

Of 299 patients evaluated for infertility between July 1, 1974, and December 31, 1976, 21 (7.2%) became pregnant during the initial evaluation prior to any recognized therapy. A comparative analysis was made between these 21 couples and the 223 completely evaluated infertility couples. No significant difference in sexual, psychiatric, or menstrual history was found. There was a significantly increased incidence of low coital frequency in the pregnancy group (P less than 0.01). The mean duration of infertility was also significantly less (1.6 years versus 2.9 years) in the pregnancy group. It is concluded that the majority of patients who become pregnant during an evaluation for infertility do so because of prior low coital exposure. There was no evidence in this study to support the concept that psychophysiologic factors were altered in these 21 couples during the initial clinical visits.

Sperm transport from the external cervical os to the fallopian tubes in women: a time and quantitation study.

PIP: Sperm transport was characterized on optimal, controlled, assessable conditions in this study. Subjects were women who had requested surgical sterilization. All subjects were restricted from coitus for 11-14 days preoperatively. The study was performed within 36 hours of the estradiol preovulatory peak for each subject; so surgical excision of both fallopian tubes was accomplished at predetermined times after insemination with fresh, whole, high-quality ejaculates; the cervical mucus column was also removed. Using this technique, sperms were identified in the oviduct within 5 minutes from deposition in the proximal vagina. Numbers of sperm in the oviduct were directly related to numbers inseminated. Sperm distribution was throughout the oviduct and was highest in fimbria (tubes had been divided in thirds upon excision). For 15-45 minutes after insemination, a constant level of sperm existed in the oviduct. 1 subject was studied similarly to determine the presence of sperm in endometrium, and no sperm occurred in that cavity until 80 minutes after insemination. This study also reports the total numbers of sperm present in cervical mucus and the correlation of insemination and time after insemination.^ieng

Continued clinical experience with an increasing dosage regimen of clomiphene citrate administration.

Between August 1, 1975, and July 31, 1977, 60 patients with infertility and ovulatory dysfunction were evaluated at the Naval Regional Medical Center, Oakland. Of these, 15 had incomplete work-ups or were lost to follow-up. Of the remaining 45 patients, 2 (4.5%) failed to ovulate at a maximal dose of 250 mg of clomiphene for 5 days with 10,000 units of human chorionic gonadotropin. Of the 43 (95.5%) who did ovulate, 33 conceived. Of these 33 patients, 9 (27%) required greater than 100 mg of clomiphene and/or longer than three ovulatory cycles to conceive. Of the 10 patients who did not become pregnant, additional infertility factors were present in 5. When these 5 were eliminated, the corrected pregnancy rate was 87% (33 of 38). The spontaneous abortion rate was 12.2% and there were no multiple gestations. These results would further emphasize that clomiphene can be used safely and effectively at doses greater than 100 mg or for longer than three ovulatory cycles.

Correlation of postcoital evaluation with in vitro sperm cervical mucus determinations and ureaplasma cultures.

Fifteen trials were completed in 14 couples during an infertility evaluation. The postcoital test (PCT) was accomplished in a standardized manner. Also, standardized determinations of the sperm-cervical mucus penetration test (SCMPT) with the addition of cross-testing (X-test) utilizing bovine cervical mucus (BCM) and normal donor semen (NDS) were done. Male and female samples were also cultured for Ureaplasma urealyticum using U9-B indicator broth and A-7 agar. The PCT and SCMPT agreed in 87% (13/15) of the cases. Utilizing BCM and NDS, where possible, the causative factor was the cervical mucus in 54% (7/13); semen factor in 15% (2/13); both factors in 8% (1/13); and undetermined in 23% (3/13). U. urealyticum cultures were positive in 40% (6/15) of the cases. Analysis of the results revealed: (1) significant correlation between PCT and SCMPT (P less than 0.01); (2) no significant difference among the PCT, SCMPT, and X-test, indicating that the cervical mucus was the causative factor; and (3) no U. urealyticum correlation with the PCT or the SCMPT. Thus, laboratory SCMPT and X-test correlated with the PCT, providing additional information concerning the causative factor in infertility. The U. urealyticum status in cervical mucus and semen cannot be determined from the PCT nor the SCMPT.

Assessment of sperm for cryopreservation using the hypoosmotic viability test.

In summary, the hypoosmotic viability parameter was significantly correlated with the outcome of the thawed sperm motility. The prefreeze supravital staining for sperm viability and the hypoosmotic sperm swelling test were not predictive of the thawed sperm total motility. The hypoosmotic viability parameter was not correlated to the postwarmed sperm motility after refrigeration. The results indicated that the integrity of the sperm membranes at the head were more important than the tail membrane.

Human papillomavirus gene sequences in washed human sperm deoxyribonucleic acid.

The present study demonstrated the presence of HPV gene sequences in Percoll-washed sperm cells using polymerase chain reaction primers targeting smaller gene regions. Up to 64% of the sperm specimens were shown to contain gene sequences indicative of the presence of HPV. Human papillomavirus type 16 was detected about twice as often as HPV type 18. The results suggest the possible role of sperm as a vector for HPV.

A new concept for anastomosis of the fallopian tube: tissue reconstruction with nonpenetrating, arcuate, legged clips in the rat model.

OBJECTIVE: To evaluate the titanium Kirsch-Zhu microclip microsurgical reanastomoses of the fallopian tubes. DESIGN: Compare the reanastomoses of the rat uterine horn between Kirsch-Zhu clips (Cushman Engineering, Albuquerque, NM) (group A) and conventional suture microsurgical techniques (group B). SETTING: Microsurgery Research Laboratory, Loma Linda University Medical Center, Loma Linda, California. PATIENTS, PARTICIPANTS: Fifteen SD rats Harlan (Harlan Sprague-Dawley Corp., Indianapolis, IN) were done in groups A and B and six in control group (C). MAIN OUTCOME MEASURE: Pregnancy rate, litter size, tissue of procedure, and histologic results. RESULTS: The clip technique was shorter in procedure time and resulted in equal fertility rate and litter size. Histologically, there were less granulomatous formation and histiocytic inflammation, but muscularis thinning and fibrosis were noted with the clip. CONCLUSIONS: The Kirsch-Zhu clip has potential for application to human fallopian tube reanastomoses.

White blood cells in semen affect hyperactivation but not sperm membrane integrity in the head and tail regions.

The presence of high numbers of peroxidase-positive PML in ejaculated semen significantly reduced sperm HA, an important step leading to sperm capacitation. Sperm membranes at both the head and tail regions, as assessed by the hypo-osmotic viability parameter and the hypo-osmotic sperm swelling test, respectively, were not affected by peroxidase-containing leukocytes. Sperm motility was not affected, but sperm curvilinear and straight line velocity parameters were reduced in the presence of high concentrations of leukocytes in the ejaculate. The results suggested that the effect of leukocytes on sperm was through a reduction in sperm hyperactive motility but not through alterations in the sperm head and tail membranes.

Sperm as a noninvasive gene delivery system for preimplantation embryos.

OBJECTIVE: To determine if sperm could be manipulated to be a noninvasive transport carrier for the delivery of gene fragments to the blastocyst. DESIGN: Sperm cells carrying foreign DNA fragments from human papillomavirus (HPV) types 16, 18, 31, and 33 were allowed to migrate from one end of an artificial reproductive tube and to come in contact with hatching mouse blastocysts at the other end of the tube. The blastocysts were then washed and analyzed for the presence of the foreign DNA fragments. SETTING: Clinical and academic research environment. MAIN OUTCOME MEASURE: Detection of amplified products from transferred foreign DNA using the polymerase chain reaction and primers targeted at the E6-E7 region for different HPV types. RESULTS: Polymerase chain reaction analyses showed transference of DNA HPV type 18 to the blastocysts. Not all types of DNA fragments were transferred equally. CONCLUSION: The results suggested the possibility of using sperm as a noninvasive gene delivery system for passing on gene fragments to preimplantation embryos. It was demonstrated that certain DNA fragments were easier to deliver than others, indicating the necessity for exploring all the factors involved in the mechanism of the transference process. The study also serves to highlight the possibility of unintentional transmission of viral or bacterial DNA to the developing embryo via the sperm.

Effect of a once weekly oral contraceptive upon gonadotrophin and gonadal steroid levels.

PIP: A moderately effective oral progestogen contraceptive, R-2323 (13beta-ethyl-17alpha-ethynyl-17-hydroxy-gona-4,9,11-trien,3-one), was administered weekly in doses of 2.5 mg and 5 mg to 5 subjects each for 8 weeks. In 2 of the 10 treatment cycles of 2.5 mg and 4 of the 10 treatment cycles of 5 mg, progesterone levels indicative of anovulation were obtained. In each of these cycles, an absence of the midcycle luteinizing hormone (LH) peak was noted. In an additional 5 cycles with absence of an LH peak, ovulatory steroid patterns were present. The remaining 9 cycles showed gonadotropin and steroid patterns consistent with ovulation. It is concluded that the main contraceptive mechanism is probably other than inhibition of ovulation. Increased viscosity of the cervical mucus is considered a possible means of contraception.^ieng

Inhibition of implantation in vitro of frozen-thawed mouse embryos by the signal molecule diadenosine tetraphosphate.

The dinucleotide polyphosphate, diadenosine 5', 5'''-P(1), P(4)-tetraphosphate (Ap4A), has been identified in mammalian and non-mammalian cells as a signal molecule that initiates the process of DNA replication and cell division. The objective of this study was to determine the function of this messenger molecule in preimplantation mouse embryonic cells. Frozenthawed two-cell mouse embryos were incubated in the presence of 0, 0.1 and 1.0 mM Ap4A at 37 degrees C in moist 5% CO(2) in air mixture for 5 d. The developmental stages of the embryos in terms of hatching and implantation were evaluated. The data showed dose-dependent inhibition of blastocyst implantation; however, there were no differences observed in the number of embryos developing to the blastocyst stage. The results suggest that Ap4A neither promotes nor inhibits the development of early stage embryos except at the implantation stage, where it exerts inhibitory control.

Effectiveness of stimulated menstrual cycles and Percoll sperm preparation in intrauterine insemination.

To examine the usefulness of intrauterine insemination in women with various fertility factors, we retrospectively analyzed data from women treated during 1986 and 1987. Ninety-three patients underwent 1-11 cycles of single or double procedures, for a total of 423 inseminations in 263 treatment cycles. Twenty-six patients (28%) conceived, for a 10% total pregnancy rate per cycle, with 58% of the pregnancies resulting from double inseminations per cycle. An average of two treatment cycles was required to achieve pregnancy. Eight pregnancies (31%) occurred in spontaneous cycles, while 18 (69%) occurred in stimulated cycles. While clomiphene citrate therapy was useful in anovulatory patients, it was of no benefit in ovulatory patients being treated with intrauterine insemination. Human menopausal gonadotropin therapy was of benefit in both ovulatory and anovulatory patients when combined with intrauterine insemination. The live birth rate was higher (75%) in spontaneous cycles than in stimulated cycles (44%). Semen preparation was accomplished by sperm washing in 61% of the pregnancies and by Percoll preparation in 39%. The effectiveness of the discontinuous Percoll gradient for semen preparation for insemination was suggested by a pregnancy rate of 9% per cycle. While the mean sperm count in the pregnant group was 44 million, successful pregnancy was accomplished with a double insemination of 880,000 and 1.16 million rapidly progressive sperm in the first and second inseminate, respectively. The data confirm the important role of intrauterine insemination for the treatment of infertility.

Evaluation of the Pipelle curette for endometrial biopsy.

The Pipelle endometrial suction curette was evaluated, and its application and effectiveness were compared to those of the Novak curette for endometrial sampling during the midluteal phase. Fifty women underwent an endometrial biopsy with the Pipelle and Novak curettes. Ninety percent of the women preferred the biopsy with the Pipelle. Histologically, tissue obtained with the Pipelle was satisfactory and similar to that with the Novak. The Pipelle curette appears to be an excellent device for midluteal endometrial biopsy during an infertility evaluation.

Effect of intramuscular triamcinolone acetonide on the human ovulatory cycle.

The effect of intramuscular triamcinolone acetonide (TCA-A) on pituitary gonadotropins and ovarian hormones was studied in a normally menstruating woman. Serum levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH), as well estradiol 17-beta (E2) and progesterone (P), were determined daily in a normal "ovulatory" pretreatment cycle. A total of 160 mg of TCA-A was then administered in four injections over two and a half months. Daily serum levels of LH, FHS, E2, and P were again measured during a period beginning thirty days after the last injection of TCA-A. Cyclicity of all these hormones was absent after treatment. Both LH and FSH were suppressed in the first half of the post-treatment period when compared with the pretreatment ovulatory cycle. A potent corticosteroid such as TCA-A is apparently capable of producing anovulatory cycles in humans by disruption of cyclic pituitary gonadotropin secretion.

PIP: The effect of triamcinolone acetonide (TCA-A) im on pituitary gonadotropins and ovarian hormones was studied in a normally menstruating woman. Serum levels of luteinizing hormone (LH) and follicle stimulating hormone (FSH), as well as estradiol-17beta (E2) and progesterone (P), were determined daily in a normal "ovulatory" pretreatment cycle. A total of 160 mg of TCA-A was then administered in 4 injections over 2 months. Daily serum levels of LH, FSH, E2, and P were again measured during a period beginning 30 days after the last injection of TCA-A. Cyclicity of all these hormones was absent after treatment. Both LH and FSH were suppressed in the 1st half of the posttreatment period when compared with the pretreatment ovulatory cycle. A potent corticosteroid such as TCA-A is apparently capable of producing anovulatory cycles in humans by disruption of cyclic pituitary gonadotropin secretion.

The interpretation and significance of the fractional postcoital test.

The investigation was designed to objectively determine the clinical usefulness of the fractional postcoital test. Forty-three normal subjects had midcycle cervical mucus collection at various times after insemination. There was a significant correlation between the number of motile sperm at the internal os level and the total sperm count within the cervical mucus. The median internal os count was 15 motile sperm per high-power field with a lower 95% confidence limit of five motile sperm per high-power field. Therefore, the fractional postcoital test is useful clinically as it is a physiologic indication of sperm transport in cervical mucus.