Search

We previously devised and reported a laparoscopic resection of benign ovarian cysts. In the present paper, we report precautions and improvements in the procedure and the application of the mini-laparotomy. 1. The subjects consisted of a total of 41 cases with preoperatively diagnosed cysts. These were 17 cases with simple cysts, 15 cases with dermoid cysts, and nine cases with chocolate cysts. The present surgical procedure was performed on 34 (83%) cases. 2. One case, preoperatively diagnosed as a chocolate cyst, was diagnosed as an endometrioid adenocarcinoma. 3. Laparotomy was required in 7 cases; in four as a result of hard adhesions, in one because it was impossible to withdraw fluid, in one due to a damaged urinary bladder and in one due to endometrioid adenocarcinoma. 4. It became essential to classify the ovarian cysts into three groups according to size. 5. Mini-laparotomy was performed for two giant ovarian cysts and five ovarian cysts in pregnant women. In this procedure the goal is minimal, simple, safe and reliable surgery with results equal to those of standard laparotomy.

We designed a new surgical method for the treatment of benign ovarian cysts. We have previously reported that, with our laparoscopy-guided method, the cyst wall is pulled outside the abdominal cavity after evacuation of the cyst contents, and resection is performed by the same method as during laparotomy. The preoperative assessment and surgical findings were investigated in 16 subjects, 5 with simple cysts, 2 with para-ovarian cysts, 2 with chocolate cysts, 5 with dermoid cysts and 2 with mucinous cysts. This surgical method was applicable in 11 of the 14 patients. Laparotomy was required in the other 3, because 2 of the patients had strong adhesions and in 1 evacuation was impossible. The patients were classified into 3 grades of difficulty before surgery: Grade I; operable, Grade II; operable with special care, Grade III; laparotomy required. During surgery, the feasibility of continuing the procedure was also assessed. In this surgical procedure, through a small incision in the abdominal wall, the entire resection of an ovarian cyst is done extra-peritoneally. However, the eligibility of patients for this method needs to be very carefully examined preoperatively and reviewed throughout the procedure.

The effect of sperm passage through cervical mucus (CM) on the fertilizing capacity of human spermatozoa was examined in the in vitro fertilization system of zona-free hamster eggs. Each drop of ejaculated semen and BWW culture medium was connected by a small capillary tube filled with preovulatory CM, egg white or BWW medium under liquid paraffin oil in a plastic petri dish. After 2 hours, zona-free hamster eggs were added to the drop of BWW culture medium containing spermatozoa which had passed through the capillary tube and the mixture was incubated for various lengths of time at 37 degrees C under 5% CO2 in air. Human spermatozoa, which were washed and preincubated for 2 hours in BWW medium, were capable of fertilizing zona-free hamster eggs but needed a longer incubation time than spermatozoa which had passed through CM. Fertilization rates of spermatozoa which had passed through CM and egg white were very similar, but no fertilization occurred in the drop containing spermatozoa which had passed through BWW medium, presumably because of the contamination with seminal plasma. These results indicate that the most important role of CM may be to separate motile spermatozoa from seminal plasma components hostile to fertilization.

The sperm penetration test (SPT) in cervical mucus (CM) is an important test in screening for sperm-CM incompatibility. For a routine SPT, a sufficient amount of preovulatory CM is necessary, but the amount of human CM is often inadequate. We examined estrous bovine CM as a substitute for human CM in the SPT. Preovulatory human CM and estrous bovine CM were collected and stored frozen at -20 degrees C until use. After being thawed at room temperature, the two CM samples were compared as to their pH, spinnbarkeit and ferning patterns, and it was found that they are quite similar. In Kremer's method of SPT, sperm penetration distance, density and motility were similar in human and bovine CM for 12 hours, but thereafter sperm motility became much lower in bovine CM than in human CM. The results of SPT with spermatozoa from patients with oligozoospermia were quite similar in both CMs. When spermatozoa pretreated with antisperm antibody were used for SPT, sperm penetration was completely blocked in human CM, but no inhibition was seen in bovine CM. From these results, we conclude that bovine CM can be substituted for human CM for some, but not all, purposes.

Rat spleen cells immunized to human azoospermic semen were fused with mouse myeloma cells (P3 X 63Ag8Ul) to obtain hybridomas which produce monoclonal sperm immobilizing antibodies. Of 114 hybrid clones obtained, 10 were found to produce antibodies with sperm immobilizing activity with complement. One clone (1C4) producing the highest antibody titer was chosen for further characterization. Chromosome analysis showed that 1C4 hybridoma cells contained two types of chromosomes which were derived from rat and mouse. Rat Ig was detected in both the cytoplasma and the culture medium of the cells by direct immunofluorescent test and immunodiffusion test, respectively. Gammaglobulin fractions from the culture medium had a strong sperm immobilizing activity and showed a common single precipitin line against human seminal plasma and human milk proteins. It was proved that the monoclonal antibody was produced to No. 7 antigen and not to No. 3 antigen (lactoferrin) in human seminal plasma, both of which have a common antigenicity to human milk proteins and compose the sperm coating antigens.

Thirteen infertile patients (18 cycles) with sperm immobilizing antibodies were subjected to in vitro fertilization and embryo replacement (IVF-ER) therapy during 12 months from January to December 1985. Four patients became pregnant, two of them delivered healthy babies at term and 2 had abortions at 5 weeks and 12 weeks of gestation, respectively. In the same duration, 24 patients (28 cycles) with tubal factor and 4 patients (5 cycles) with male factor for infertility were subjected to IVF-ER, and two patients with tubal factor became pregnant and delivered healthy babies at term. In the patients with immunological factor, fertilization and cleavage rates per mature oocyte were 85.9% (55 fertilized/64 oocytes) and 81.3% (52 cleaved/64 oocytes) respectively, while the fertilization rates in patients with tubal factor and male factor were 70.5% (62 fertilized/88 oocytes) and 50.0% (6 fertilized/12 oocytes) respectively and all fertilized eggs in these patients developed to the cleavage stage. Thus fertilization and cleavage rates for mature oocytes from the patients with the immunological factor were slightly better than those with the tubal factor and much better than those with the male factor. The antisperm antibody titers (SI50) in sera determined by the quantitative sperm immobilization test ranged from 20 to 243 units while those in follicular fluids ranged from 21 to 160 units in the patients with the immunological factor. The follicular sperm immobilizing antibodies could be detected in any patient who had the antibodies in the serum. Immunoglobulin (IgG, IgA, IgM) concentrations in follicular fluids were not significantly different from each other in the patients with immunological, tubal and male factor for infertility.

In order to apply the early pregnancy factor (EPF) to early diagnosis of fertilization, the establishment of optimal conditions for assay of EPF was attempted, and then EPF in the sera of contracepted and conceived women 4 to 6 days after ovulation were measured. For assay of EPF, 0.25 ml of 1:2 step diluted anti-human lymphocyte serum (ALS), 0.05ml of guinea pig serum as complement and 0.1ml of lymphocytes suspension (1 X 10(7)/ml) pretreated with test serum were mixed and then incubated at 37 degrees C for 90 min. To this mixture 0.1ml of sheep red blood cell suspension (2 X 10(9)/ml) was added and the rosette formation was counted after centrifugation. The rosette inhibition titer (RIT) was expressed as reciprocal of ALS dilutions which resulted in less than 75% of rosette formation as compared with the control. RITs of the conceived women who were assayed on the 5 th day after ovulation were in the range from 16 to 32 X 10(3), while that of the control contraceptive women who were assayed on the same day was in the range from 2 to 4 X 10(3). The sterile women who received AIH but failed to conceive all showed less than 4 X 10(3) as RIT. These results suggest that the assay of EPF is valuable in detecting the early stage of fertilization and possibly may help to differentiate the impairment of embryo implantation from non-fertilization of the ovum as a cause of sterility.