HLA-DR and HLA-DQ gene typing of infertile women possessing sperm-immobilizing antibody.

Thirty-eight infertile women, possessing sperm-immobilizing antibody (SIA), were examined for their HLA-DR and -DQ types using DNA obtained from peripheral blood cells. The typing of HLA-DR and DQ was performed by polymerase chain reaction sequence specific oligonucleotide probes (PCR-SSOP) and PCR-restriction fragment length polymorphism (RFLP), respectively. In comparison to the normal Japanese population, the SIA positive patient population had higher genes frequencies in HLA-DRB1*0901 (26.3 vs. 13.6%, P<0.005), DQB1*0602 (13.2 vs. 6.2%, P<0.05) and -DQB1*0303 (26.3 vs. 14.8%, P<0.01), but not in any HLA-DQA1 gene types by chi2 test. After Bonferroni correction, the high frequency of HLA-DRB1*0901 remained significant (P<0.05) and HLA-DQB1*303 was slightly significant (P<0.07) but no other genes had a gene frequency significantly higher than that of the normal Japanese population. HLA-DRB1*0901 and HLA-DQB1*0303 are very rare among Caucasians but characteristically high among Japanese. The high frequency of HLA-DRB1*0901 and DQB1*0303 genes in the Japanese population may account for higher frequency of sperm-immobilizing antibody in Japanese compared to other ethnic groups.

Prenatal sonographic diagnosis of a fetal renal mesoblastic nephroma occurring after transfer of a cryopreserved embryo.

Here we report the first case of prenatally diagnosed fetal renal mesoblastic nephroma occurring after transfer of a cryopreserved embryo. A 37 year old woman, having immunological infertility, was treated by in-vitro fertilization (IVF) and embryo transfer. Following unsuccessful IVF using fresh embryos, the patient conceived after transfer of cryopreserved-thawed embryos. The chromosomal analysis identified a normal karyotype at 16 weeks' gestation when amniocentesis was performed. The pregnancy course was uneventful until 28 weeks' gestation when polyhydramnios associated with fetal renal tumour was detected using ultrasonography. A male infant weighing 2564 g was born via Caesarean section at 34 weeks' gestation. A left nephrectomy was performed 5 days after delivery and the tumour was identified histologically as a mesoblastic nephroma. The postoperative course was uncomplicated to this point.

Relationship between human in-vitro fertilization and intracytoplasmic sperm injection and the zona-free hamster egg penetration test.

The zona-free hamster egg penetration test (HEPT) is widely used for evaluating the fertilizing ability of human spermatozoa. However, the relationship between the HEPT and microassisted fertilization has yet to be determined. To evaluate the efficiency of HEPT in selecting the most appropriate method of in-vitro fertilization (IVF), including intracytoplasmic sperm injection (ICSI) in couples with male factor infertility, clinical laboratory data was analysed retrospectively. The patients were divided into groups according to the sperm penetration index as determined by the HEPT: group A (sperm penetration index = 0), group B (sperm penetration index < 15) and group C (sperm penetration index > or = 15). A total of 405 oocytes were collected and inseminated by conventional methods in 69 couples with male factor infertility. In all, 31 out of 148 (20.9%) oocytes fertilized in group A; 35 out of 117 (29.9%) in group B; and 73 of 140 (52.1%) in group C. The clinical pregnancy rates per transfer in groups A, B and C were 0% (0/13), 0% (0/14) and 25.9% (7/27) respectively. Both the fertilization rate and pregnancy rate in group C was significantly higher than in groups A and B. ICSI was carried out in a total of 57 couples and 334 oocytes in metaphase II stage were manipulated. The normal fertilization (2 pronuclear) rate per oocyte was 65.6 +/- 26.0% (mean +/- SD). Out of 127 oocytes, 76 (59.8%) fertilized in group A, 57 out of 87 oocytes (65.5%) in group B and 86 out of 120 oocytes (71.7%) in group C. Of the 56 transfers, 17 clinical pregnancies were obtained, giving an average pregnancy rate of 30.4% per transfer. The clinical pregnancy rates per transfer in groups A, B and C were 17.4% (4/23), 40.0% (4/10) and 39.1% (9/23) respectively. No significant differences were observed in the fertilization rates or in the pregnancy rates between the three groups. In addition, there were no differences in the fertilization and pregnancy rates between the ICSI and IVF patients in group C. These findings suggest that the results of the HEPT are well correlated with the fertilizing ability of human spermatozoa in the patients treated by conventional IVF. Couples suffering from male factor infertility with a sperm penetration index of < 15 (as determined by HEPT) should consider treatment with ICSI, while those with a sperm penetration index of > or = 15 should attempt conventional IVF.

Effect of troglitazone on blood insulin levels after pancreas transplantation with systemic venous drainage in rats.

BACKGROUND: Troglitazone is a new oral antidiabetic agent and has been reported to reduce insulin resistance and improve peripheral hyperinsulinemia in patients with noninsulin-dependent diabetes mellitus. To examine the effect of troglitazone on insulin regulation after pancreas transplantation with systemic venous drainage, we measured peripheral glucose and insulin levels and performed an intravenous glucose tolerance test. METHODS: We divided the rats into four groups: diabetic rats with a pancreas graft and administration of troglitazone at 40 mg/day orally (group P+T, n=4), rats with a pancreas graft only (group P, n=4), age-matched normal rats (group N, n=5), and diabetic rats (group DM, n=4). RESULTS: Fasting insulin levels in group P were relatively higher than those in group N, whereas the values in group P+T were normalized. In the intravenous glucose tolerance test, troglitazone clearly regulates sigma immunoreactive insulin levels of pancreas transplanted rats (P vs. P+T: 244+/-23 vs. 145+/-14 microU/ml, P<0.05). CONCLUSION: Hyperinsulinemia induced by systemic venous drainage, which may progress atherosclerosis, can be controlled with troglitazone treatment.

A case of a patient diagnosed with malignant mixed Müllerian tumor of the ovary who conceived after conservative surgery and adjuvant chemotherapy.

A case of successful pregnancy after treatment of stage Ia malignant mixed Müllerian tumor of the ovary is described. This tumor is very rare and usually occurs in postmenopausal women. cis-platinum-based chemotherapy following the primary operation in the early stage is the most effective treatment, although most of the tumors have been found in the advanced stage. In this patient, treatment with unilateral salpingo-oophorectomy to preserve fertility was performed and followed by cis-platinum-based chemotherapy for 5 years. Two years after completion of the chemotherapy, she spontaneously conceived and the course of the pregnancy has been uneventful to this point.

Evaluation of sperm fertilizing ability using the Sperm Quality Analyzer.

The Sperm Quality Analyzer is an inexpensive device which provides a quantitative estimation of sperm motility. To evaluate the fertilizing ability of human spermatozoa using a Sperm Quality Analyzer, correlations amongst the sperm motility index, the sperm penetration index (as assessed using the sperm penetration assay; SPA), and the fertilization rate in the treatment of standard IVF-ET were analysed retrospectively. The sperm motility index demonstrated a significant correlation with sperm concentration (p < 0.001), sperm motility (p < 0.001) and the motile sperm concentration (p < 0.001) in a total of 104 fresh semen samples from 81 men donating samples for IVF-ET. The sperm motility index also showed a significant correlation (p < 0.001) with the sperm penetration index in 60 patients, assessed using the SPA, before they were treated by standard IVF-ET. The correlation between the sperm motility index and the IVF-ET fertilization rate was higher than that between the sperm penetration index and the fertilization rate. The sperm motility index was classified into three categories: 'poor' (sperm motility index < 80), 'medium' (sperm motility index 81-160) and 'good' (sperm motility index > 160). The relationships between the IVF-ET fertilization rate and each category of the sperm motility index values were also evaluated. For the three categories in the sperm motility index, the fertilization rates (76.0%) of 60 samples judged as 'good' were significantly higher than those (44.2%) of 15 samples judged as 'medium' (p < 0.001) and those (34.7%) of 13 samples judged as 'poor' (p < 0.001). These results indicate that the Sperm Quality Analyzer provides a reliable estimation of the fertilizing ability of human spermatozoa.

Characterization of anti-sperm antibodies and their coding cDNA sequences by Epstein-Barr virus transformed B cell lines from lymphocytes of infertile women possessing anti-sperm antibodies.

Epstein-Barr virus (EBV)-transformed B cell lines that produce human antisperm antibodies were established using peripheral blood lymphocytes (PBLs) from infertile women with sperm immobilizing antibodies in their sera. We obtained three stable cell populations (designated B1, B2, D5) of transformed PBLs originating from three different patients. They produced IgM sperm-reacting antibodies directed against the tail of live, methanol-fixed and NaIO4-treated human spermatozoa. The established antisperm antibodies recognized noncarbohydrate sperm membrane antigens with different specificity and distribution in the male reproductive system. Antisperm antibody-B2 corresponding antigen appears to be specific for the male reproductive system. This antigen is excreted from the epithelial cells of the ductus epididymidis and bound to the spermatozoa in the lumen of the ductus. Antisperm antibodies B1 and D5 corresponding antigens were expressed on the spermatozoa in the seminiferous tubules and were common to the secretions of the ductus epididymidis, prostate and some other somatic organs. The cDNA of the immunoglobulin heavy chain genes were analyzed by reverse transcription polymerase chain reaction (RT-PCR) using RNA extracted from these clones. The immunoglobulin heavy chain cDNA sequences of these antisperm antibodies showed extremely high homology to previously reported immunoglobulin germline DNA sequences, implying that these antisperm antibodies might be natural autoantibodies rather than antibodies stimulated by external antigen.

A successfully treated case of multiple liver abscesses accompanied by portal venous gas.

A case with multiple liver abscess accompanied by massive portal venous gas is reported. A 61-yr-old male was admitted because of left lower abdominal pain, fever, and diarrhea. Abdominal x-ray examination demonstrated multiple branching lucencies in the liver. Computed tomography revealed multiple liver abscesses and massive gas in the portal system as well as a thickened wall of the sigmoid colon. Enema study using contrast medium revealed a perforation of the sigmoid colon with diverticulitis. The outcome was favorable after sigmoid colectomy in addition to intensive treatment with antibiotics. Bacteroides fragilis, which produces gas (H2 and NH3) by fermentation, was isolated not only from the resected specimen but also from blood samples. Although the presence of portal venous gas is a sign of poor prognosis in patients with intestinal infectious diseases, the sensitive detection of hepatic portal venous gas by computed tomography and the appropriate treatment may improve the patient's prognosis.

Effects of sperm immobilizing antibodies on pregnancy outcome in infertile women treated with IVF-ET.

PROBLEM: Since it was found that anti-sperm antibodies could impair in vitro development of fertilized eggs in the presence of complement in rats, the effects of sperm immobilizing antibodies on human pregnancy were examined in infertile women treated with IVF-ET. METHODS: The pregnancy outcomes of 143 ET cycles in 58 infertile women with sperm immobilizing antibodies and 363 ET cycles in patients with tubal infertility as control were compared. Diagnosis of chemical pregnancy was done when the urinary hCG level had risen over 50 IU/L but a gestational sac could not be demonstrated later. Antibody titers of sperm immobilizing antibodies (SI50 units) were measured by a quantitative sperm immobilization test. RESULTS: 33 (23.1%) of 143 cycles in the patients with sperm immobilizing antibodies and 56 (15.4%) of 363 cycles in the control patients were diagnosed as pregnancy. The pregnancy rates were significantly higher in the former than in the latter (P < 0.05). In the patients with sperm immobilizing antibodies, 12 (36.4%) were chemical pregnancies, 5 (15.2%) were clinical abortions, and 16 (48.5%) had deliveries. In the control group, 18 (32.1%) were chemical pregnancies, 10 (17.9%) were clinical abortions including ectopic pregnancies and 28 (50.0%) had deliveries. There was no significant differences in each category. When the SI50 titers at the time of conception were considered, chemical pregnancy rates were 22.2% (4/18) in patients with SI50 titers below 10 units, but those in patients with SI50 titers above 10 were 50.0% (5/10) and above 100 were 60.0% (3/5), respectively, (P > 0.05). In four of five patients who had both chemical and clinical pregnancies, the SI50 titers at the time of conception were higher in the chemical pregnancy cycles than in the clinical pregnancy cycles. CONCLUSIONS: Though the pregnancy rates were significantly higher in the patients with sperm immobilizing antibodies as compared to those with tubal infertility, chemical pregnancy rates were also higher in the patients with higher sperm immobilizing antibody titers. These results suggest that sperm immobilizing antibodies may cause the damage of early development of human embryos in vivo in the small number of patients with a high titer of the antibodies.

The role of pancreatic islets in experimental pancreatic carcinogenicity.

Our previous studies have suggested that the presence of intact islets is essential for the induction of pancreatic exocrine tumors in the Syrian hamster model. To validate this, we investigated the effect of the carcinogen, N-nitrosobis(2-oxo-propyl)amine (BOP) in hamsters, in which homologous isolated intact islets were transplanted into the submandibular gland (SMG). Freshly isolated pure islets from hamster donors were transplanted into the left SMG of 20 female host hamsters. Ten of these hamsters (group 1) received BOP (40 mg/kg) weekly for 3 weeks. Another 10 hamsters (group 2) were kept untreated. In groups 3 and 4 (10 hamsters each) the salt solution or isolated pancreatic ductal cells, respectively, was injected into the gland. In other groups (10 hamsters each) islets were transplanted into the peri-SMG connective tissue (group 5) or into the renal subcapsular space (group 6). Hamsters of group 1 (40 mg/kg, weekly for 3 weeks) as were group 7 hamsters, which served as BOP-treated controls. All BOP-treated hamsters developed pancreatic lesions. Similar hyperplastic and atypical ductal/ductular proliferation and in situ carcinoma were found in the SMG of many group 1 hamsters. No such lesions were found in the SMG, peri-SMG, or renal subcapsular space of the other groups. Islets appear to be involved in carcinogenicity of BOP. The mechanism is obscure.

[Maternal serum alpha-fetoprotein as a screening test for fetal Down syndrome].

An association between a low maternal serum alpha-fetoprotein (MSAFP) level and an increased incidence of Down syndrome is well documented. This study was undertaken to elucidate the diagnostic efficacy of MSAFP as a screening test for Down syndrome in 711 Japanese women who underwent genetic amniocentesis between 14 and 20 gestational weeks of age in Hyogo Medical College Hospital. The mean value for the multiple of the median (MoM) of the MSAFP level in 11 pregnancies associated with Down syndrome was 0.78 +/- 0.47 and 1.12 +/- 0.67, respectively. A cut-off value of < 0.45, < 0.50, < 0.55 and < 0.60 MoM yielded negative predictive values of 98.6%, 98.5%, 99.0% and 99.2%, positive predictive values of 6.3%, 4.0%, 9.1% and 8.6%, a sensitivity of 9.1%, 9.1%, 36.4% and 54.5%, and false positive rates of 2.1%, 3.4%, 5.7% and 9.1%, respectively. The combining of MSAFP and maternal age improves the expected results of a strategy for Down syndrome.

Insulin release from a bioartificial pancreas using a mesh reinforced polyvinyl alcohol hydrogel tube. An in vitro study.

Islet transplantation with a bioartificial pancreas is a potential alternative to whole pancreas transplantation. The authors constructed a bioartificial pancreas using mesh reinforced polyvinyl alcohol hydrogel tubes (MRPT), in an attempt to clarify the in vitro responsiveness to glucose of islets seeded in the MRPT. When the MRPT were perfused in a small chamber with buffer containing 3.3 mmol or 16.7 mmol glucose, insulin release from the MRPT began to increase at 9 +/- 3 min, reaching a plateau at approximately 40 min after the glucose concentration in the perfusate increased from 3.3 to 16.7 mmol. When MRPT seeded with islets were subjected to static incubation in buffer containing 3.3 mmol or 16.7 mmol glucose, insulin release from the MRPT remained elevated for 3 hr of high glucose stimulation, the amount of secreted insulin depending upon the number of islets seeded. Although pre incubation of semipermeable membranes in culture medium containing fetal bovine serum prior to seeding with islets has recently been reported to improve insulin release, the authors found that such pre treatment of the MRPT did not have a beneficial effect. Their in vitro findings in this study suggest that the bioartificial pancreas using MRPT could be a promising therapeutic approach to human diabetes mellitus.