Feasibility and efficacy of intraperitoneal docetaxel administration as salvage chemotherapy for malignant gynaecological ascites.

Ovarian and endometrial cancers diagnosed at advanced stages are often associated with malignant ascites. This study aimed to determine the safety, feasibility and efficacy of intraperitoneal (IP) docetaxel (TXT) for the treatment of ascites. A phase I study, including nine patients, was undertaken to determine the maximum tolerable dose. Efficacy was retrospectively assessed in 18 patients treated with 40-70 mg/m(2) IP TXT between 2005 and 2012. In a phase I study, the dose was safely escalated to a maximum of 70 mg/m(2), at which level no patients had grade -3 haematological adverse events. In a retrospective study of 18 patients, seven had an Eastern Cooperative Oncology Group performance status of 3; 16 had prior paclitaxel administration and two, with doses of 40 and 70 mg/m(2), experienced a serological response and a decrease in paracentesis. Thus, palliative treatment of recurrent OC should be further studied with 40 mg/m(2) among more patients, and 70 mg/m(2) could be evaluated for first-line IP chemotherapy.

Mechanism of palmitic acid-induced deterioration of in vitro development of porcine oocytes and granulosa cells.

The concentration of fatty acids in follicular fluid reflect the physical condition of donors, and palmitic acid (PA) is a major component of follicular fluid. The present study examined the effect of PA on in vitro oocyte growth and investigated the molecular backgrounds of the PA induced-low quality oocytes. Oocyte-granulosa cell complexes (OGCs) were collected from early antral follicles of gilts. The OGCs were cultured for 14 days in a medium containing 0.5 mM PA or vehicle (BSA). PA was found to reduce granulosa cell (GCs) proliferation (0.73 fold) and viability (93.9% vs. 85.8%) and increase lipid content in oocytes and GCs. Oocytes developed in the presence of PA had low developmental ability to the blastocyst stage. In addition, PA affected developmental and epigenetic markers of histone modifications in oocytes; levels of H4K12 acetylation and H3K9 demethylation. PA affected cellular proliferation, apoptosis and endoplasmic reticulum stress markers along with reducing the phosphor-AKT/AKT levels and increasing the expression levels of caspase-3 and CHOP in GCs. Incubation of OGCs with PA increased ceramide content in the GC, and addition of ceramide to the culture medium inhibited GC proliferation. In conclusion, it is suggested that high PA content in the medium reduces viability and proliferation through ceramide accumulation, and PA impaires the developmental ability of oocytes grown in vitro. In addition, high-fat conditions induce changes in the histone modifications of oocytes grown in vitro.

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.

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.

Effects of sperm-immobilizing antibodies on sperm-zona pellucida tight binding.

OBJECTIVE: To evaluate the in vitro effects of sperm-immobilizing antibodies on sperm-zona pellucida (ZP) tight binding. DESIGN: The hemizona assay (HZA) was used to study the inhibitory effects of infertile women's sera with and without sperm-immobilizing antibodies on sperm ZP tight binding. These results were compared with those of monoclonal sperm-immobilizing antibodies. SETTING: The patients were collected from a university hospital infertility clinic. PATIENTS: Sera from 40 infertile women (24 with and 16 without sperm-immobilizing antibodies) and 2 postpartum women as control were used. RESULTS: Of 24 patients' sera with sperm-immobilizing antibodies, 23 (96%) showed significant inhibitory effect, whereas none of 16 patient's sera without sperm-immobilizing antibodies exhibited any inhibitory effect. However, there was no correlation between the antibody titers of sperm-immobilizing antibody and the hemizona index. Among four monoclonal sperm-immobilizing antibodies tested, only one showed a significant inhibitory effect on the sperm-zona tight binding. A human monoclonal antibody derived from an infertile woman with sperm-immobilizing antibodies, whose serum showed an inhibitory effect on HZA, did not inhibit the HZA. CONCLUSIONS: There are at least two kinds of sperm-immobilizing antibodies, one with both activities of sperm immobilization and blocking of sperm-zona tight binding and another with the former activity alone. The vast majority of sperm-immobilizing antibodies reduce zona binding even without the presence of complement.

A pilot study of nedaplatin and etoposide for recurrent gynecological malignancies.

Nedaplatin is a new analogue of cisplatin with similar efficacy but less renal toxicity. We assessed the efficacy and toxicity of nedaplatin and etoposide for recurrent gynecological malignancies. Eight patients were treated with 100 mg/m2 of nedaplatin (day 1) and 70 mg/m2 of etoposide (days 1, 3, and 5 ) every 4 weeks. A total of 17 courses was given. Grade 3/4 leucopenia and thrombocytopenia occurred, but was manageable. The response rate was 37.5% (partial response n=3; no change n=4; progressive disease n=1). In conclusion, a phase II study of this regimen for recurrent gynecological malignancies is warranted.

A case complicated with SLE during maintenance hemodialysis.

A 40-year-old woman suffered from toxemia of pregnancy in 1977 and was admitted to hospital. Thereafter, she developed nephrotic syndrome, underwent a renal biopsy, and a diagnosis of membranoproliferative glomerulonephritis (MPGN) was made. She received steroid therapy, immunosuppressive drug and anticoagulant therapy, and recovered sufficiently to be discharged from hospital in April, 1979. During subsequent ambulatory treatment at our outpatient department, her renal function deteriorated gradually, and maintenance hemodialysis was started from June, 1990. In July, 1991, she was admitted to our hospital with pleurisy and pericarditis. There was no improvement despite antibiotic treatments. Laboratory data revealed leukopenia and lymphopenia. Under suspicion of systemic lupus erythematosus (SLE), relevant tests were carried out. Immunological abnormalities such as positive LE cells and the presence of various autoantibodies, together with clinical signs of hypersensitivity to sunlight, stomatitis and serositis, satisfied the diagnostic criteria of the ARA and a diagnosis of SLE was made. This case did not exhibit any clinical or serological abnormalities except for the renal disorder for a 10-year period after the histological diagnosis of MPGN, but was eventually diagnosed as SLE as a result of the manifestation of SLE symptoms for the first time after one year of maintenance hemodialysis. Immunological abnormalities and SLE during maintenance hemodialysis are discussed in relation to other reports.

High resolution electron microscopic observation of hydroxyapatite in tooth crystals.

Structure images taken with a high resolution electron microscope with a resolution of 0.18 nm revealed that the atomic arrangement in tooth-enamel hydroxyapatite (HA) corresponds with the calculated image using crystallographic data from mineral HA (Ca10(PO4)6(OH)2). Direct information on the two types of Ca and the OH- groups in the HA crystals was obtained from the observed image through [001] projection. The established correspondence between the observed image contrast and the arrangements of the columnar Ca, the screw-axis Ca, and the OH- groups suggests that HREM observations are useful to identify defects in HA crystals.

[Laparoscopic resection of ovarian cysts: precautions and improvements in the procedure and application of the mini-laparotomy].

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.

[Laparoscopy-guided extraperitoneal resection of ovarian cysts: surgical procedure, indications and limitations].

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.

Sperm immobilizing antibodies interfere with sperm migration from the uterine cavity through the fallopian tubes.

PROBLEM: It is well known that sperm migration in cervical mucus is impaired by sperm immobilizing antibodies secreted in the mucus. However, it is not clear yet whether sperm migration from the uterine cavity through the fallopian tubes to the peritoneal cavity is impaired by sperm immobilizing antibodies. To test the possible impairment of sperm migration in the tubes, laparoscopic examinations were carried out and the presence of motile sperm in the peritoneal fluid after intra-uterine insemination was investigated. METHOD: Peritoneal sperm recovery tests were performed in 28 infertile women with sperm immobilizing antibodies in their sera, and the results were compared with those in 322 infertile women without the antibodies. Both the sperm immobilizing antibody titers (SI50) and complement activities (C'H50) in peritoneal fluid were compared with those in patients' sera. In some experiments, the supernatant of the peritoneal fluid was used as a source of complement for the sperm immobilization tests instead of guinea pig serum. RESULTS: Among couples with normal semen characteristics by the criteria of WHO, sperm recovery in the peritoneal fluid was observed in only 3 (11.1%) of 27 patients with sperm immobilizing antibodies, compared with 72 (34.0%) of 212 patients without the antibodies (P < 0.025). The antibody titers of the patients with the sperm recovery were very low by the quantitative sperm immobilization test. In most patients, a similar amount of sperm immobilizing antibodies was present in the peritoneal fluid and the sera. Though the complement activities in the peritoneal fluid were less than those in sera, the former were still found to be sufficient to immobilize sperm in vivo. CONCLUSIONS: These results suggest that the complement-dependent sperm immobilizing antibodies could interfere with sperm migration in the female genital tract at the level of the fallopian tubes.

Assay of testicular angiotensin-converting enzyme activity in human spermatozoa.

The testicular isozyme of angiotensin-converting enzyme (ACE) is associated with male fertility. Spermatozoa from mice lacking ACE showed defects in transport within the oviducts and in binding to zonae pellucidae although the animals had normal sperm count, morphology and motility. In fact, unexplained infertility is difficult to be predicted by conventional parameters such as sperm count. We measured membrane testicular ACE activity in a sperm suspension in PBS and total testis ACE activity in spermatozoa by solubilization with Triton X-100. Total testis ACE activity and membrane testis ACE activity of the same subject were compared in 12 control subjects. We demonstrated that testicular ACE is stable in spermatozoa and the assay of testicular ACE activity is possible. Total testicular ACE activity was approximately twice the membrane testicular ACE activity in all of the subjects tested. The assay of testicular ACE activity in human spermatozoa could be a new method for the assessment of sperm function.

Prediction of ovarian reserve based on day-3 serum follicle stimulating hormone concentrations during the pituitary suppression cycle using a gonadotropin releasing hormone agonist in patients undergoing in vitro fertilization-embryo transfer.

Satisfactory results following in vitro fertilization-embryo transfer (IVF-ET) treatments depend on retrieving an appropriate number of mature oocytes without causing the development of ovarian hyperstimulation syndrome (OHSS). The present study was carried out to investigate whether the ovarian reserve is predictable based on the day-3 serum concentration of follicle stimulating hormone (FSH) during the pituitary suppression cycle using a gonadotropin releasing hormone (GnRH) agonist (defined as day-3 FSH) in patients undergoing IVF-ET treatment. Day-3 FSH before the administration of gonadotropin was assessed in 72 IVF-ET cycles from 59 infertile women. The mean+/-SD of day-3 FSH, the total amount of FSH plus human menopausal gonadotropin (hMG) administered, and the total number of oocytes retrieved was 5.5+/-2.6 mIU/ml, 2834.2+/-1236.5 IU and 7.7+/-5.8, respectively. There were significant correlations between day-3 FSH and the total amount of FSH-hMG administered (p < 0.001), and day-3 FSH and total number of oocytes retrieved (p < 0.001). There was a significant difference of day-3 FSH between patients who subsequently conceived (4.4+/-1.3 mIU/ml) and those who did not conceive (6.1+/-2.9 mIU/ml) (p = 0.001). There was also a significant difference of day-3 FSH between patients who developed moderate or severe OHSS (4.5+/-1.2 mIU/ml) and those who did not (5.9+/-2.8 mIU/ml) (p = 0.003). Receiver-operator characteristic curve analysis showed that the significant cut-off point for day-3 FSH for predicting ovarian reserve was 5.25 mIU/ml. These findings indicate that day 3-FSH is usefulfor predicting ovarian reserve during the pituitary suppression cycle using a GnRH agonist in patients undergoing IVF-ET.

Inhibition of sperm-zona pellucida tight binding by sperm immobilizing antibodies as assessed by the hemizona assay (HZA).

PIP: Researchers collected serum samples from 23 infertile patients with sperm immobilizing antibodies (SI-Ab) and 1 pregnant patient from the Department of Obstetrics and Gynecology at the Hyogo Medical College in Japan to screen sera to determine whether they contained factors to inhibit sperm-zona pellucida tight binding. They used the recently developed hemizona assay (HZA) to test for this binding. The HZA assay showed that all 23 serum samples inhibited sperm-zona pellucida tight binding. The hemizona index (HZI) ranged from 3-53 with a mean of 18.1 (standard deviation of = or - 12) compared to a normal (HZI) of 100. Serum samples with titers 10 of SI50 inhibited sperm-zona binding as well as those with titers -or= 10 of SI50 (HZIs=17.3 vs. 18.9; p.1). All 23 serum samples bound to the surface of sperm plasma membrane after 1 hour coincubation as evidenced by the fact that they all demonstrated 50% IgG beads bound. Further the results of the indirect immunobead test (I-IBT) showed that positive sera (+or= 20% IgG beads) significantly inhibited binding more than negative sera (20% IgG beads bound) (HZIs=12.4 vs. 24.4; p.05). Yet serum with positive I-IBT for IgM did not affect sperm-zona binding (HZIs=17.1 vs. 19.4; p.1). No association existed between HZI and site of IB binding. The researchers interpreted theses results to mean that sera with both SI-Ab and antibodies recognized I-IBT for IgG and IgA may play a significant role to inhibit the sperm-zona pellucida tight binding. In conclusion, physicians should expect patients with low HZI to have more problems conceiving than those with normal HZI. In vitro fertilization using heat inactivated human cord serum or donor serum may help them to conceive.^ieng

High-resolution electron microscopy of high-temperature superconductors.

We review recent results obtained at Northwestern using high-resolution electron microscopy to study high-temperature superconductors. While in general these materials form large, very perfect single crystal grains which display very few imperfections, there is also evidence of slip defects, amorphous regions, and order-disorder transformations. We also report that the gadolinium-based superconductors and in one case yttrium-based superconductors show evidence for some copper solid solubility in the form of copper-rich planar defects. The structure of a metastable trigonal polytype is also reported, as are the effects of electron beam and water vapor damage to the materials.

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.

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.