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The present study was undertaken to assess the role of tumor infiltrating lymphocytes and polymorphonuclear leukocyte-derived human leukocyte elastase (HLE) as an inactivator of urokinase-type plasminogen activator (uPA) activity. We have investigated the localization of pro-uPA/uPA and HLE immunohistochemically and quantitated in paraffin-embedded formalin-fixed uterine cervical cancer tissue sections (stage II) in 13 patients with positive lymph-nodes metastasis and in 16 cases with node-negative. The invasively growing and metastasizing tumor cells consistently contained uPA enzyme activity. When investigated immunohistochemically with antibody to uPA, different parts of individual tumors showed a pronounced heterogeneity in staining intensity. Strong staining was found in the areas with invasive growth and degradation of surrounding normal tissue. A statistically significant higher node-positive rate was observed in patients having tumors with strong uPA stainings than in those with weak stainings. On the other hand, inflammatory cells such as polymorphonuclear cells were located in aggregates or diffusely spread within tumor stromal tissue. The inflammatory reaction seemed to be most intense at the border between the surrounding normal stromas and the tumor tissue. These cells produced and secreted HLE. A statistically significant higher node-positive rate was observed in patients having tumors with weak HLE stainings than in those with strong stainings. Thus, uPA and HLE staining intensities in tissue specimens appear to be independent predictors of increasing and decreasing risk for node-positive, respectively, suggesting that HLE may act as a defence against tumor cell invasion and metastasis.

High molecular mass glycoproteins detected by the monoclonal antibody OC125 were found in secreted and solubilized materials from the ovarian cancer cell line, HOC-I. The CA125 antigen has been isolated from HOC-I cell extract and conditioned media by perchloric acid precipitation, gel filtration, and OC125 affinity purification. The higher molecular mass complexes (molecular masses were estimated to be > 1,000KDa) was predominantly found in extracts of cells grown in vitro or in ovarian cancer tissues, whereas the lower molecular mass antigen (molecular masses were estimated to be 110-400KDa) was the major component in conditioned media and sera from patients with ovarian cancer. Double determinant enzyme-linked immunosorbent assays were carried out with several antibodies (OC125, NS19-9, TKH-2, B72.3, CC49, MA54, MA61, PC47H) immobilized on a solid phase to bind different epitopes on antigens in clinical samples. Biotinylated OC125 was used as a probe to detect the CA125 antigen that had been bound via different epitopes. When TKH-2 was used to bind the antigen and OC125 used as a probe, the coexpression of CA125/sialyl Tn was observed in 12(22%) of 55 sera from patients with ovarian cancer. In some cases, sialyl Tn or CA19-9 antigens may be present on the CA125 macromolecular complexes.

Sialyl Tn structure (NeuAc alpha 2-6 GalNAc 1-0-Ser/Thr) recognized by monoclonal antibody TKH-2 is a characteristic component in meconium- and/or amniotic fluid-derived mucin. The purpose of this study was to evaluate whether immunohistochemical staining employing antibody TKH-2 is a sensitive method for histological diagnosis of amniotic fluid embolism (AFE). The intravenous injection of 10% meconium solution (1 ml/kg) into rabbits was carried out. Formalin-fixed, paraffin-embedded lung, liver, and kidney tissue sections were obtained at various time interval after injections. Specimens were stained by the Streptavidin Biotin Complex method. The results of immunostaining were compared to those of hematoxylin and eosin (H & E) stain or a conventional Alcian blue stain. Remarkable positive stainings were easily seen within the pulmonary artery vasculature in experimental rabbits. Meconium-derived mucin can be easily missed on H & E sections. Compared to TKH-2 staining, Alcian blue staining may also be insufficient to demonstrate intravascular mucin in the lung sections. TKH-2 staining is the most sensitive method to use in detecting meconium-derived mucin in the lung sections of rabbits with an AFE experimental model.

The fibrin adhesion technique is being recognized as a way to prevent amniotic fluid leakage in the case of preterm premature rupture of the fetal membrane (PROM). However, the effect of fibrin adhesion is still not clear. The aim of this study is to investigate the implications of fibronectin in repairing the fetal membrane. We divided 9 normal fetal membranes into 4 groups. In all the groups, an incision was made in the membrane with a sterile knife. Group A: The incisions in the membranes were sealed with fibrinogen and thrombin (N = 9). Group B: The incisions in the membranes were sealed with fibrinogen and thrombin with fibronectin (N = 9). Group C: Fibronectin was added to the culture medium containing fetal membranes (N = 9). Group D: The fetal membranes were cultured in the medium alone (N = 9). All membranes were cultured for 6 days in 10% fcs added to MEM and analyzed histologically. In group A, fibrin clots did not attach to the membranes at all. In group B, fibrin clots attached closely to the membranes. In group C, fibroblasts increased remarkably at the incision sites on the fetal membranes. In group D, fibronectin accumulated at the incision sites on the fetal membranes. These results suggest that fibronectin is important in repairing fetal membrane.

Five monoclonal antibodies (moABs TKH-2, MA54, MA61, B72.3, and CC49), directed toward the O-linked mucin-type glycoprotein, showed signs of specific reactivity with human meconium. The reactivity of these moABs with meconium extract was examined by solid-phase ELISA with different native and sialidase-treated glycoproteins. All moABs react with meconium extract, whereas the reactivities of TKH-2, MA54, and MA61 are sialidase sensitive and the reactivity of TKH-2 with meconium extract was only inhibited by ovine submaxillary mucin (OSM), indicating that TKH-2 is the most sensitive and specific antibody clearly directed to the sialyl Tn antigen in meconium. The possible application of TKH-2 to diagnose amniotic fluid embolism (AFE) has been prelimiarily investigated. We demonstrated that the concentration of sialyl Tn antigen in the serum of patients with AFE was significantly increased, indicating that meconium was released into the maternal circulation. Our method for detecting sialyl Tn antigen in the serum of AFE patients is a direct way to demonstrate the release of meconium into the maternal circulation, and is a simple, rapid, non-invasive and sensitive method for the diagnosis of AFE.

We have investigated the expression of epidermal growth factor receptor (EGF-R) in invasive cervical cancer tissue of the uterus to determine whether there is a relationship between the expression of EGF-R and lymph node metastasis. Frozen sections of surgical specimens from twenty patients with invasive cervical cancer were immunohistochemically stained by the alkalinephosphatase anti-alkalinephosphatase method. The monoclonal antibody 528 we employed reacts with the receptor binding epitope within the EGF molecule, indicating that monoclonal antibody 528 is competitive with ligands such as EGF. Immunohistochemical staining was carried out before and after dissociation of ligands from the cellular surface of the tumor by acid treatment. However, the staining was resulted in no difference between before and after acid treatment. Consequently, the expression of EGF-R was detected in all cancer tissues as well in some normal tissue such as basal cell layers of epidermis. The expression of EGF-R was related inversely with lymph node metastasis by Wilcoxon rank sum test (p less than 0.05). These findings suggest that the expression of EGF-R in cervical cancer does not always lead to tumor growth or tumor invasion, although it does in some types of cancer.

We purified CA125 antigen from the conditioned media (CM) of eutopic and heterotopic endometrial epithelial cells (EC) as well as ovarian cancer cell lines, SHIN-3 and HOC-I, to determine what molecular weight forms of CA125 antigen were identifiable. Treatment of the high-molecular-weight CA125 antigen with 6M urea yielded a much lower molecular mass peak. After purification by OC125 affinity column chromatography, samples were applied to 3 to 7% polyacrylamide gradient gel and analyzed by Western blot. A single band with a molecular weight (MW) of 200KDa was identified in eutopic EC materials. The CA125 polypeptide of the 110KDa molecule could be detected in all of the CM obtained from heterotopic EC, irrespective of the length of time in the cell culture. A MW of approximately 200 KDa was also observed in some heterotopic EC samples. On the other hand, although the multiple bands with a MW equal to orless than 200KDa were observed in the CM of two ovarian cancer cells, the CA125 polypeptide of 110KDa molecules could not be detected. This preliminary finding offers promise that the 110KDa molecule detection method may be a useful adjunct in the differential diagnosis of heterotopic EC and ovarian cancer.

Cervical maturation, dilatation and uterine contraction in imminent premature delivery are closely related to chemical mediators from activated granulocytes which infiltrate into the cervix. It is known that urinastatin (urinary trypsin inhibitor, UTI) inhibits many kinds of chemical mediators from granulocytes and macrophages such as granulocyte elastase (elastase) and interleukin 1. We examined the effect of a UTI suppository on uterine contraction and the elastase level in cervical mucus in cases of imminent premature delivery. We treated 43 cases of imminent premature delivery with tocolysis index 3 or 4 with 4 kinds of therapy: Group A (N = 12): ritodorine drop infusion therapy; Group B (N = 9): daily UTI suppository (1,000U) therapy; Group C (N = 14): daily UTI suppository + ritodorine drop infusion therapy; Group D: daily UTI suppository + ritodorine drop infusion + antibiotics (oral cepharosporine) therapy. The elastase level of cervical mucus before treatment was 0.76 +/- 0.40 micrograms/ml in group A, 0.93 +/- 0.43 micrograms/ml in group B, 0.85 +/- 0.40 micrograms/ml in group C and 0.90 +/- 0.41 micrograms/ml in group D. There was no significant difference between these groups. The elastase level in cervical mucus was 0.75 +/- 0.47 micrograms/ml in group A, 0.27 +/- 0.35 micrograms/ml in group B, 0.27 +/- 0.33 micrograms/ml in group C and 0.30 +/- 0.19 micrograms/ml in group D, respectively. The elastase level was decreased significantly in groups B, C and D. The time taken to depress uterine contraction was 65 +/- 66 min in group A, 375 +/- 336 min in group B, 70 +/- 64 min in group C and 58 +/- 53 min in group D, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

We have established an in vitro system in which epithelial cells (EC) and stromal cells (SC) from the human endometrium are placed in culture to examine the production of CA125. There was a much greater amount of CA125 in heterotopic EC after culture cells reached confluence than during the logarithmic growth phase. Heterotopic EC in culture secreted nine times as much CA125 constitutively after reaching confluence as eutopic EC. Flow cytometry (FCM) analysis indicated that the changes in CA125 expression correlated with the cell cycle. The CA125 expression was mainly observed in the G0/G1-phase in the cell cycle. There was no amplification of the CA125 expression in heterotopic EC membranes. The results of SDS-PAGE and Western blot indicated that the 110 KDa molecule of CA125 might be specific for adenomyosis. The biochemical and physical nature of CA125 was examined to characterize the antigenic determinant of this antigen. These results strongly suggested that the CA125 antigenic determinant from EC was composed of conformationally dependent peptides. We conclude that significantly increased secretion of CA125 from heterotopic EC could be attributed to the increase in serum CA125 in patients with adenomyosis.

Six tumor-associated antigens, cancer antigen 125 (CA125), tissue polypeptide antigen (TPA), ferritin (Fr), carcinoembryonic antigen (CEA), alpha fetoprotein (AFP), and sialyl Lex-i (SLX) were measured simultaneously for the early detection of ovarian cancer. To decrease the number of both false positive and false negative cases in the combination assay, statistical discrimination analysis employing the serum values for appropriate tumor markers has been studied with respect to ovarian cancer by the method of Mahalanobis' generalized distance. The new "ovarian cancer screening test" designed by us has been used in Shizuoka Prefecture since 1988, and 23,307 serum samples have been analyzed. One hundred twenty-seven of 165 ovarian cancer patients were suspected as having cancer by such clinical procedures as pelvic examination and/or ultrasonography, while in 150 patients cancer was detected by the statistical discrimination method. Thirty-one of 38 patients with ovarian cancer overlooked by the clinical procedures could be found by the statistical method. We conclude that clinical procedures and the statistical method can be complementary in detecting patients with this malignancy.

Circulating serum sialyl Tn (STN) antigen levels were measured in 89 patients with epithelial ovarian cancer, 157 benign disease, and in 126 healthy controls. Serum antigen levels were increased in 48.3% of patients with ovarian cancer. The false positive rate is significantly low (4.0% in healthy controls and 9.6% in benign disease). The levels of STN antigen were significantly higher in sera of patients with cancer than in those in benign and healthy controls (p less than 0.05). The rise in serum STN antigen levels correlated to the size of the primary tumors. Of the histological type, it is interesting to note the high sensitivity in mucinous-type ovarian cancer. Survival at 1, 2, 3, 4 and 5 years for patients with STN-negative (serum STN levels less than 50 U/ml) versus STN-positive (serum STN levels greater than or equal to 50 U/mol) was 96.2, 92.3, 86.5, 82.7, and 76.9% versus 59.5, 29.7, 18.9, 10.8, and 10.8%, respectively (p less than 0.05). The overall survival probability was worse in patients with STN-positive sera. Percent progression-free survival at 1, 2, 3, 4 and 5 years for patients with STN-negative versus STN-positive was 90.4, 86.5, 76.9, 59.6, and 51.9% versus 35.1, 16.2, 8.1, 8.1, and 5.4%, respectively (p less than 0.05). The overall progression-free period of survival was shorter in patients with STN-positive sera. Multivariate regression analysis revealed that positive STN, stage, PS and histologic grade were the four variables of most importance in predicting survival. These results indicate that a positive STN antigen level in sera is an independent predictor of poor prognosis in ovarian cancer.

Granulocyte elastase activity in cervical mucus (elastase activity) was measured in pregnant women (10-20 gestational weeks) with cervical polyp and controls (gravida with no cervical polyp). Elastase activity in the cervical polyp group was 86 +/- 44 U/l, and that in the control group was 22 +/- 13 (p less than 0.01). Elastase activity after polypectomy was 44 +/- 24. It decreased significantly compared with prepolypectomy (p less than 0.05). A high level of elastase activity continued in cases without polypectomy. Immunohistochemical staining of elastase for cervical polyps was also performed. In the low elastase staining group there was low elastase activity (46 +/- 26 U/l). Elastase activity in the moderate staining group was 89 +/- 31, and that in the intense staining group was 114 +/- 31. As for the prognosis of patients with cervical polyps, the occurrence rate of chorioamnionitis is 9% in the control group, 14% in the polyp with polypectomy group and 40% in the polyp without polypectomy group. There was a significant difference between the polyp without polypectomy group and other groups (p less than 0.01). These data suggest that cervical polyp is a focus of inflammation and that it may cause chorioamnionitis. We conclude that cervical polyp during pregnancy should be removed. If polypectomy could not be performed, local anti-inflammatory and anti-infectious therapy would be needed.

We studied the localization of adhesive proteins at the site of implantation in mouse pregnancy (5 days' gestation), and human pregnancy (5-8 weeks' gestation) and in human endometrial culture cells. Immunohistochemical staining of adhesive protein such as fibronectin (FN), laminin, type IV collagen (CIV), subunit A of factor XIII (XIIIA), and subunit S of factor XIII (XIIIs) were performed by indirect enzyme immunoassay to investigate their localization. (1) In mouse pregnancy: FN and XIIIs were stained in trophoblastic giant cells, and CIV was stained in the endometrial epithelium. (2) In human pregnancy: (fetal side) CIV was stained diffusely on the surface of syncytiotrophoblast. FN, CIV, and XIIIA were stained in the stroma of the villi. (maternal side) FN, CIV, and XIIIA were stained in the endometrial stroma. (3) In endometrial culture cells: FN and XIIIA were stained strongly in the stromal cells. These findings demonstrated that FN and XIIIA are produced in the endometrial stromal cells, and suggest that FN, CIV, and XIIIA are concerned with the attachment of the embryo to the uterus. Therefore, these adhesive proteins are important factors in maintaining pregnancy.

Autoimmune disorders such as SLE and ITP occur more commonly in young women and are the most common complications in pregnancy. There is considerable controversy concerning the risk to the mother and fetus, and the optimal prepartum management for minimizing that risk. 1. SLE is an autoimmune disorder in which IgG antibodies such as anti dsDNA-IgG, anticardiolipin IgG, and anti SS-A/Ro IgG are produced. Lupus nephropathy accompanied by diminished serum complement (CH50) and a rise in antibodies against dsDNA is a frequent clinical problem during pregnancy, which represents the adverse effect of hypertension or superimposed toxemia and causes fetal death or intrauterine fetal growth retardation. Habitual abortion or fetal death is common in a case with high anticardiolipin IgG titre. Anti SS-A antibodies are often found in the infants of antibody-positive mothers, and the deposition of antibodies in the perinodal region cause congenital heart block. IgG or immune complexes crossing the placenta directly injures the cardiac conduction system. In these cases which have high titre crossing the placenta directly injuries the cardiac conduction system. In these cases which have high titre of autoimmune antibodies, corticosteroid therapy should be started. 2. Management of ITP in pregnancy involves the consideration of three issues: 1) treatment of maternal thrombocytopenia, 2) prediction of fetal thrombocytopenia, 3) obstetrical management. ITP increases the risk for postpartum bleeding of sufficient severity to require blood transfusion. In most of these cases, maternal platelet counts are found to be less than 30,000/mm3. Women who have symptomatic severe steroid-unresponsive ITP may benefit from intravenous IgG(IvIgG) given as elective treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Porphyrins are found in fetal urine, neonatal urine, amniotic fluid, and meconium. Zn-coproporphyrin (Zn-CP), which we identified in meconium, has two fluorescent peaks at 580 nm and 630 nm, and Coproporphyrin (CP), found in fetal urine, has one fluorescent peak at 620 nm, when both porphyrins are excited at 405 nm. In this study, the fluorescent peak at 580 nm was used to detect Zn-CP in neonatal urine, using the new index, that is the Urinary Fluorescence Meconium Index (UFMI); UFMI = F580-(F580 + F600)/2 [F: fluorescence intensity]. We obtained urine samples from neonates in these three groups: Group-I: Meconium Aspiration Syndrome (MAS; n = 3), Group-II: non-MAS with meconium stained amniotic fluid at delivery (n = 18), and Group-III: non-MAS with clear amniotic fluid at delivery (n = 25). The UMI (Urinary Meconium Index) was also measured by conventional optical absorbance. The results show that many false positive cases were observed in UMI, and that no false positive cases were observed in UFMI. These results lead one to speculate that Zn-CP, which is specific in meconium, is used in UFMI measurement, while UMI measurement by absorbance cannot exclude contamination by CP in neonatal urine, so Zn-CP and CP are measured all together. UFMI values reflected the clinical course of neonates. It is concluded that UFMI is a more sensitive method than UMI for the diagnosis of MAS.

The remission rate of choriocarcinoma has greatly improved since the introduction of effective multiagent chemotherapy combined with aggressive surgical therapy and radiotherapy. In addition to these, the registration and follow-up of gestational trophoblastic disease (GTD) also has been playing an important role in the early detection and treatment of choriocarcinoma following hydatidiform mole. The system for the registration and follow-up of GTD was started in 1977 in Shizuoka Prefecture. In the present series, the results obtained with this registration and follow-up system from 1977 to 1988 in Shizuoka Prefecture were reviewed and analysed. 1) One thousand, nine hundred and twenty-five cases of hydatidiform mole, 68 cases of invasive mole, 70 cases of persistent trophoblastic disease and 48 cases of choriocarcinoma were registered in 12 years. The overall registration rate was 97.4%. 2) The number of cases of hydatidiform mole registered has decreased from about 180 cases to about 140 cases per year, probably due to the decreasing birth rate. 3) The number of cases of choriocarcinoma registered has recently been decreasing significantly and the number of cases of registered invasive mole and persistent trophoblastic disease has decreased slightly. 4) Antecedent pregnancy with choriocarcinoma including clinical choriocarcinoma has been changing from "post-molar" to "post-term" in the past 12 years. The prognosis of the patient with choriocarcinoma following hydatidiform mole has improved by the early detection and treatment since the introduction of the registration and follow-up system. More attention should be paid to choriocarcinoma following term gestation not yet included in the registration and follow-up system to facilitate early detection and treatment.

We measured five tumor markers simultaneously for serodiagnostic testing as a method for the early detection of ovarian cancer. To decrease both false negativity and false positivity in the results of combination assay, statistical analysis including factor analysis and stepwise discriminating function was applied in this study. At least one of these tumor markers was detected as positive in 75.2% (76 of 101 patients) of sera from patients with ovarian cancer before treatment. Six hundred and ninety-three of 7,097 normal sera (9.8%) gave spuriously positive combination assay results. Falsely positive combination assay results were observed in 1,107 of 3,139 patients with benign disease, which could largely be attributed to the high CA125 values in patients with endometriosis. On the basis of factor analysis in order to decrease false positivity, CA125, TPA, and CA125/TPA were selected as the best parameters for distinguishing between ovarian cancer and benign conditions including those pelvic endometriosis showing positive results in combination assays. Subsequently, the function derived by factor analysis made possible the correct classification of 58 of 71 patients with ovarian cancer detected by combination assay, and 84.8% of pelvic endometriosis and benign ovarian tumor subjects were correctly classified into the non-cancer group. Next, in order to decrease false negativity, statistical analysis was also applied. Malignancy could be correctly diagnosed by this procedure in 14 of 25 patients whose tumor was undetectable by combination assay, whereas of subjects without cancer 14.5% were errorenously classified into the ovarian cancer group. In the search for the best method for accurately detecting ovarian cancer, we used image diagnosis (ultrasonography) in combination with serological diagnosis.(ABSTRACT TRUNCATED AT 250 WORDS)

We studied whether the administration of vitamin K to mothers could increase the concentration of vitamin K in breast milk and prevent idiopathic vitamin K deficient bleeding in breast-feeding infants. Sixty puerperal women were divided into three groups, the control group, Menaquinone-4 (MK-4) administered group and vitamin K1 administered group. We measured the concentrations of vitamin K1, MK-4 and MK-7 in maternal plasma and breast milk on the fourth day after delivery. In the MK-4 group, the concentrations of MK-4(2.13 ng/ml in plasma, 49.3 ng/ml in milk) were significantly higher than in the control group (0.28 ng/ml, 1.51 ng/ml). In the vitamin K1 group, the concentrations of vitamin K1 (49.0 ng/ml in plasma, 71.6 ng/ml in milk) were significantly higher than in the control group (1.17 ng/ml, 2.41 ng/ml). The concentration rates (milk/plasma ratio) of vitamin K1, MK-4 and MK-7 were 2.52, 5.43 and 0.52 in the control group, 1.60, 40.2 and 0.67 in the MK-4 group and 1.65, 10.8 and 0.71 in the vitamin K1 group, respectively. The concentration rate of MK-4 was higher than that of vitamin K1 and was increased by MK-4 administration. After delivery, the daily concentration of MK-4 in milk was increased from 1.69 ng/ml on the first day to 49.3 ng/ml on the fourth day in the MK-4 group. These results indicate that MK-4 is accumulated and concentrated into breast milk, and continuous MK-4 administration can increase the concentration of vitamin K in milk, preventing idiopathic vitamin K deficient bleeding in infants.

The clomiphene or hMG to stimulate ovarian follicle maturation which we call "controlled ovarian stimulation (COS)", in vitro capacitation (IVC) of spermatozoa and artificial insemination of husband's sperm (AIH) were simultaneously given to refractory infertile couples. The following couples were studied in 5 groups, 1 male subfertility 27 cases, 2 functional infertility 25 cases, 3 ovulatory dysfunction due to central mechanism 8 cases, 4 cervical mucous insufficiency 3 cases, 5 endometriosis 6 cases. The pregnancy rate was 21.7% in total (15 of 69 cases): 25.9% in group 1, 16.0% in group 2, 25.0% in group 3, 33.3% in group 4 and 16.7% in group 5. The follicle size and serum estradiol were measured every day during the periovulatory phase to evaluate follicle maturation. The Percoll centrifugation method was compared with the washing-centrifugation method with respect to the recovery rate and the concentration rate of motile spermatozoa. The recovery rate was 67.9% with the Percoll centrifugation method, and 40.0% with the washing-centrifugation method. The concentration rate was 1.81 times in the Percoll centrifugation method, and 1.19 times in the washing-centrifugation method. The present studies indicated that the combination of COS and IVC methods for AIH in refractory infertile couples was very effective.

The usefulness of tumor markers in serodiagnosis of cancer designed to detect ovarian cancer at an early stage was evaluated from the point of view of their diagnostic value. Namely, eight tumor markers, CA125, SLX, CA72-4, TPA, Fr, CEA, CA19-9, and SCC, were determined and studied to find the combination that would yield the optimal diagnostic value. For this purpose, the diagnostic value was calculated from sensitivity x specificity. As a single tumor marker CA125 proved optimal with a diagnostic value of 0.50. The higher value, 0.53, was obtained as the diagnostic value from the combination of two tumor markers, CA125 and CA72-4. When three tumor markers were combined, CA125, CA72-4 and SLX gave an optimal diagnostic value of 0.65. In the combination of four tumor markers, CA125, CA72-4, SLX and CA19-9 gave a diagnostic value of 0.63. In the five marker combination CA125, CA72-4, SLX, CA19-9 and TPA worked well and had a diagnostic value of 0.59. When the markers were increased to six types, CA125, CA72-4, SLX, CA19-9, TPA and Fr provided a combination with 0.53 as the diagnostic value. In the seven marker combination CA125, CA72-4, SLX, CA19-9, TPA, Fr and CEA gave a diagnostic value of 0.51. The efficiency declined to 0.51 when eight tumor markers were combined. When cost performance in the measurement of tumor markers for early detection of ovarian cancer is taken into account, a dilemma arises in that the increase in the number of tumor markers used is accompanied by higher sensitivity and lower specificity.(ABSTRACT TRUNCATED AT 250 WORDS)

Anticancer chemotherapy with cisplatin (CDD) as the main drug (combined with adriamycin (ADM) and cyclophosphamide (CPM), PAC therapy) was performed on patients with ovarian cancer. Urinastatin (US) was concurrently administered to assess its effectiveness in preventing CDDP-induced nephrotoxicity. Twenty-two patients with gynecological malignant tumor were treated with PAC therapy, and of these, twelve concurrently received US. The ten who did not receive US served as the control. As a rule, one course of PAC therapy consisted of 50mg/m2 CDDP, 50mg/m2 ADM and 500mg/m2 CPM. Before the administration of CDDP, US 100,000 units was administered by I.V. drip infusion and after the administration, US 400,000 units was again administered by I.V. drip infusion at a speed of 100,000 to 200,000 units/hour. A total of approximately 3,500ml of fluids was administered I.V.. Each course of PAC therapy took 7 to 14 hours to complete. The control group underwent PAC therapy in a regimen not including US. As indexes of nephrotoxicity, serum levels of BUN, creatinine (Cr), and creatinine clearance (Ccr), and N-acetyl-beta-glucosaminidase (NAG), gamma-glutamyl transpeptidase (gamma-GTP), and arylamidase (AA) activity in the urine was determined before treatment and at days 1, 2, 3, 7, 14, and 21 after the initiation of PAC therapy. Changes in serum BUN, Cr, and Ccr levels after CDDP administration in the group with and the group without concurrent US were similar. Urinary gamma-GTP, AA, and NAG activity remained unchanged after CDDP administration in the group with concurrent US. In contrast, in the group without US, this urinary enzyme activity was transiently increased after CDDP administration.(ABSTRACT TRUNCATED AT 250 WORDS)

We measured the six tumor markers, CA125, TPA, Fr, CEA, AFP, and PPA simultaneously in serodiagnostic testing as a method for the early detection of ovarian cancer. To decrease the false negativity in the combination assay, statistical discriminating analysis was applied to serum values for tumor markers. "The ovarian cancer screening test" designed by us has been enforced in Shizuoka Prefecture since September, 1985. Twenty-one thousand nine hundred serum samples had been analysed by December, 1988. Eighty-four cases out of the 125 patients with ovarian cancer were suspected of being malignant pretherapeutically in a clinical diagnostic procedure which included echography. On the other hand, malignancy was justified before treatment in 109 patients in the combination assay and statistical analysis. One hundred patients were diagnosed as malignant in the combination assay and 9 cases by statistical analysis. However, we overlooked 4 patients (3 patients were found to be in stage I and one in stage III, respectively) even when both methods were used. Altogether 37 more patients with ovarian cancer could be found by the serodiagnostic screening method with discriminating analysis using tumor markers than by clinical findings such as image diagnosis and others. To conclude, the discriminating analysis was, therefore, verified as a useful technique for the diagnosis of ovarian cancer.

The plasminogen activator exists in the human placenta (PPA) as a reversible complex with its inhibitor (UKI). Plasminogen activating activity appeared resulting from the separation of UKI from PPA-UKI complex through UK-Sepharose or UK-Affi Gel 10 affinity chromatography. This crude PPA was purified through gel filtration on Sephadex G-150 and DEAE-Sephacel column chromatography. The purified PPA was obtained at a rate of 25 micrograms from one placenta, the specific PA activity of which was 21, 071 IU/mg-protein. The molecular weight of PPA was estimated at about 65,000 (unreduced) by SDS-polyacrylamide gel electrophoresis. PPA did not react to the anti-UK serum and UK did not react to the anti-PPA serum. It is revealed that PPA has a smaller influence on fibrinogenolysis than UK does. PPA is a new plasminogen activator in the human placenta, which has different properties from UK biologically and immunologically. It is speculated that in the near future PPA will be one of the tissue activators in thrombolytic therapy for thrombotic diseases including toxemia. We note that the measurement of the plasma PPA level is useful as one of the marker proteins of placental function and malignant tumors.

We succeeded for the first time in extracting and purifying a coagulation fibrinolysis factor in the placenta, that is, a placental plasminogen activator (PPA). Using the enzyme-labeled antibody technique (indirect method), we investigated the localization of PPA, fibrinogen (FBG) and fibronectin (FN) in the placenta. And we tried to elucidate the physiological significance of the above three. The F(ab')2 fragments of their antibodies were produced by pepsin digestion, because there were various Fc receptors in the placenta. The results are summarized as follows: PPA was produced in the trophoblast, and was secreted out of the microvilli. FBG was located in the plasma. But, in the placenta, it was not in the cells and the connective tissue of which chorionic villi was composed. FN was located in the ground substances of connective tissue in the chorionic villi, and was associated with the cell structure. Because PPA, FBG and FN showed the same distribution on the fibrinoid material, it is expected that coagulation fibrinolysis activity proceeds in that fibrinoid material.

The efficacy of two methods of chemotherapy in the treatment of patients with advanced ovarian cancer was compared on the basis of survival curves; one consisted of remission induction therapy alone with a combination of cisplatin (CDDP), adriamycin (ADM) and cyclophosphamide (CPM) (induction PAC therapy), and the other consisted of induction PAC therapy and additional maintenance therapy with cyclic PAC (cyclic PAC therapy). The subjects of the study were patients with advanced ovarian cancer in stages III and IV. Sixty-eight patients received induction PAC therapy alone and seventeen patients received both induction and cyclic PAC therapy. Demographic factors such as age at initial presentation, the stage of cancer (III or IV), surgical procedure, histological classification, number of courses of induction PAC, response rate, site of residual tumor after surgery and induction PAC therapy, and reduction rate of CA125 were compared in the two groups. When analyzed by the chi 2 test, none of these factors was significantly different in the two groups. Patients in the induction PAC therapy group received a median total dosage of CDDP 360 mg, ADM 235mg, and CPM 2.246mg. Patients in the cyclic PAC therapy group received CDDP 592mg, ADM 490mg, and CPM 4.642mg. Thus, the dosage of anticancer agents administered to the latter group was about twice as great as that administered to the former group. According to the Kaplan-Meier method, survival rates for the induction PAC therapy group were 88.2% for one year, 50.0% for two years, 28.0% for three years, and 8.8% for five years. The median survival period was 23 months.(ABSTRACT TRUNCATED AT 250 WORDS)

The authors recently studied the histocompatibility between patients with choriocarcinoma and their husbands and did an in vitro experiment (in an autosystem) to establish whether choriocarcinoma can be treated with lymphocytes from the husband. The results were as follows. Seventeen pairs of trophoblastic diseases were analyzed for HLA antigens. The histocompatibility rate between the patient and husband was 60% in the choriocarcinoma group, which was larger than the rates for both the hydatidiform mole group (29%) and the invasive mole group (0%). We succeeded in establishing a cultured cell strain of choriocarcinoma. We named this strain SMT-cc 1 and used it in the subsequent autologous experiments. Peripheral T-lymphocytes of the patient with SMT-cc 1 were examined for their cytotoxicity against SMT-cc 1 using the 51Cr release assay. T-lymphocytes of the patient showed a cytotoxicity of 7.6% after independent culture (control experiment), while they showed a cytotoxicity of 19.7% after a 6-day mixed culture with the husband's lymphocytes. The latter figure was almost equal to the rate (22.2%) observed for T-lymphocytes after a similar stimulation procedure using SMT-cc 1. When anti-HLA-A, B antibody was added during the above mentioned cytotoxicity test, cytotoxicity was markedly reinforced. The results of this study indicate the effectiveness of specific immunotherapy which, making use of the fact that choriocarcinoma expresses the husband's genome, intravenously injects lymphocytes into the patient, after stimulation with antigens (husband's lymphocytes or choriocarcinoma cells).