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To explore the pathogenesis in placental dysfunction and abruptio placentae, we analyzed the occurrence of placental cell apoptosis and the role of Fas and Fas ligand (L) in that process in an inflammatory placental dysfunction model of pregnant mice, using lipopolysaccharides (LPS). In the present study, Day 13 pregnant mice were injected i.p. with LPS (50 microg/kg) or saline as a control, and the placentas were isolated at various time points after the injection. Analysis of the isolated DNA in agarose-gel electrophoresis revealed a typical ladder pattern of bands consisting of 180-200 base pairs (bp), which is regarded as a hallmark of apoptosis. The intensity of the bands increased time-dependently, reaching a maximum level at 12 h after LPS injection. In accord with the biochemical data, histochemical analysis using terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) revealed that nuclei positive for double-stranded DNA breaks were found in decidua, diploid trophoblasts in the basal zone, and spongiotrophoblasts. The number of positive nuclei was maximized at 12 h after LPS injection. As a next step, we investigated the possible involvement of Fas and Fas L in the induction of apoptosis of the placental cells after LPS injection. Western blot analysis indicated that LPS increased the expression of Fas and Fas L in the placenta by about 4-fold at 12 h and 18 h, respectively, after injection. The cells expressing Fas and Fas L were identified, using immunohistochemistry and nonradioactive in situ hybridization, as decidua, diploid trophoblasts in the basal zone, and spongiotrophoblasts. Furthermore, when the expression of 4-hydroxy-2-nonenal (HNE)-modified proteins was assessed to evaluate the relation of oxidative stress elicited by LPS to the induction of apoptosis, once again decidua, diploid trophoblasts in the basal zone, and spongiotrophoblasts were positive. Therefore, the placental dysfunction by LPS may be brought about by the Fas-mediated apoptosis of various placental cells in a paracrine/autocrine fashion, possibly under the influence of oxidative stress.

We have previously shown that thioredoxin and thioredoxin reductase were immunohistochemically localized in cytotrophoblasts, decidua and stromal cells in the stem villi of human placenta and that the addition of exogenous thioredoxin and thioredoxin reductase to mitochondrial fractions from human placenta displayed a protective effect on fumarase activity against oxidative stress. In this study, to investigate further the roles of thioredoxin and thioredoxin reductase in protecting pregnancy against oxidative stress, we examined the effect of lipopolysaccharide (LPS), which induces a variety of cytokines and produces radical oxygen species, on the expression of thioredoxin and thioredoxin reductase in mouse placenta. We focused on the placental protective effect in the second trimester, when the onset of placental dysfunction might occasionally lead to a critical state for the fetus. Thus we analysed placentae from mice on day 13 of pregnancy at various time points after they were injected with LPS (50 microg/kg i.p.) or saline as a control. The expressions of thioredoxin and thioredoxin reductase were evaluated by Western blotting and immunohistochemistry. Western blot analysis revealed that LPS approximately quadrupled the expression of both thioredoxin and thioredoxin reductase in the placentae of pregnant mice. When both proteins were localized immunohistochemically, it was found that the decidua and the diploid trophoblasts in the basal zone were intensively stained. Furthermore, the expression of 4-hydroxy-2-nonenal (HNE)-modified proteins, which are markers of oxidative stress, was enhanced in placenta by LPS. Our study suggests that the induced thioredoxin and thioredoxin reductase might protect the placenta from the stress induced by LPS., (Copyright 1999 Harcourt Publishers Ltd.)

The purpose of the present study was to investigate the correlation between tumor size and prognosis in stage IB and II cervical cancer and to elucidate the adequacy of new FIGO staging system for cervical cancer. The subjects included 128 patients with cervical cancer (stage IB = 86, IIA = 18, and IIB = 24) who had undergone radical hysterectomy. The largest tumor size of the pathology specimen was measured in two dimensions, and the correlation between tumor size and prognosis was investigated. In addition, tumor size of the pathology specimen was compared with the largest tumor diameter measured by MRI in stage IB cancers. Patients with a tumor size greater than 3 cm2 had a significantly worse 5-year survival rate (63%) when compared to those with tumor size no greater than 3 cm2 (96%) (P < 0.0001). Multivariate analysis revealed that independent prognostic factors were tumor size (P = 0.003) and lymph node metastasis (P = 0.015). By regression analysis, the largest tumor size of the pathology specimen was relatively well correlated with the largest tumor diameter by MRI in stage IB cancers; 3 cm2 of tumor size in the pathology specimen corresponded to 3.4 cm of tumor diameter by MRI. The adequacy of new FIGO staging system was considered relatively acceptable.

Objective: To examine whether or not estrogens induced the expression of protein thiol/disulfide oxidoreductases such as protein disulfide isomerase (PDI), thioredoxin (Trx), Trx reductase, and glutaredoxin (Grx) in vascular endothelial cells., Methods: The regenerative effects of the protein thiol/disulfide oxidoreductases, PDI, Trx and Grx, on oxidatively damaged proteins were assayed using H2O2-inactivated glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a reporter enzyme. The induction of protein thiol/disulfide oxidoreductases and the accumulation of protein adducts generated by lipid peroxidation were examined by Western blotting in estrogen-treated bovine aortic endothelial cells (BAECs)., Results: Reduced PDI, Trx and Grx regenerated the H2O2-inactivated GAPDH in vitro. The levels of these protein disulfide oxidoreductases in BAECs were increased by pretreatment with 0.01-10 micromol/l 17beta-estradiol, the largest increase (about fourfold of the control) being found for PDI. Other sex hormones such as progesterone and testosterone did not affect the contents of these oxidoreductases in BAECs. 4-Hydroxy-2-nonenal (HNE)-protein adducts, which are generated by lipid peroxidation, were accumulated in BAECs exposed to paraquat, whereas the pretreatment of BAECs with 17beta-estradiol suppressed their accumulation., Conclusions: The estrogen-mediated induction of the protein thiol/disulfide oxidoreductases such as PDI, Trx, Trx reductase and Grx suggested a possible involvement of these oxidoreductases in the antioxidant protection of estrogen observed in the vascular system.

Background: Cytologic findings of sarcoma botryoides were still equivocal because sarcoma botryoides of the uterine cervix is an extremely rare neoplasm, and few cases have been reported to date., Case: A 17-year-old female was diagnosed with sarcoma botryoides of the uterine cervix. The entire vaginal canal was occupied with polypoid masses, which arose from the anterior lip of the uterine cervix, and the tumor was classified as group I (Intergroup Rhabdomyosarcoma Study). After wedge resection and six courses of combination chemotherapy, the tumor recurred in the same location of the cervix as the primary lesion. Touch smear of the polypoid mass formed loose clusters and also showed short spindle cells in a necrotic background. The nucleus of the tumor cells had a thin nuclear membrane, fine chromatin pattern and partly clear nucleolus, showing mild nuclear atypia. Immunohistochemically, some of the tumor cells showed positive staining for myoglobin and desmin., Conclusion: The cytologic findings of sarcoma botryoides of the female genital tract are typical features of nonepithelial malignant tumor. Immunohistochemical study is useful for the diagnosis of rhabdomyosarcoma.

Two groups of intermediate-to high-grade non-Hodgkin's lymphoma patients aged 65 years and over were enrolled in a dose-range study of CHOP therapy utilizing 5 doses (1/2, 7/12, 2/3, 5/6, full CHOP): 11 patients 65-79 years of age (group A) and 9 patients 80 years or older (group B). The patients were enrolled consecutively; the study was designed so that if 3 patients completed 3 cycles of CHOP on schedule without major problems, the next highest dose was administered. If 2 patients experienced any major problems during 6 cycles at a given dose, treatment was discontinued and the dose prior to that particular dose was regarded as the optimal dose. The 6 treatment cycles were completed by 3 of 3 (2/3 CHOP), 3 of 4 (5/6 CHOP), and 1 of 4 (full CHOP) group A patients; and by 3 of 3 (1/2 CHOP), 2 of 3 (7/12 CHOP) and 1 of 3 (2/3 CHOP) group B patients. The results indicated that the optimal doses for group A and B were five-sixths and seven-twelfths of the standard CHOP dose, respectively.

Recent studies have indicated that oxidative stress is involved in the pathogenesis of pre-eclampsia. Oxidative stress damages systemic tissues, and placental damage may result in intrauterine growth retardation and fetal distress. Thus, this study attempted to elucidate the placental localization of thioredoxin and thioredoxin reductase, substances that may reduce oxidative stress. Furthermore, it studied the defence mechanism of the thioredoxin-thioredoxin reductase system against oxidative stress in mitochondria of normal human placenta where reactive oxygen species are primarily produced. The examination of thioredoxin reductase activity in subcellular fractions of human placenta indicated that thioredoxin reductase was located not only in cytoplasm, but also in mitochondria. The existence of thioredoxin and thioredoxin reductase in human placenta was confirmed immunologically using antibodies raised against thioredoxin and thioredoxin reductase. Thioredoxin and thioredoxin reductase were localized histochemically in cytotrophoblasts, decidua, and stromal cells in the stem villi. The addition of exogenous thioredoxin and thioredoxin reductase to fumarase in mitochondria of human placenta displayed a protective effect against oxidative stress. In conclusion, this study confirmed the intracellular localization and the tissue distribution of thioredoxin and thioredoxin reductase in human placenta. Moreover, the complete thioredoxin-thioredoxin reductase system in human placenta may protect the placenta from damage caused by oxidative stress.

Objective: To investigate the correlation between the development of low grade cervical intraepithelial neoplasia (LCIN) and human papillomavirus (HPV) infection in cases with long-term follow-up., Study Design: Forty-three cases of LCIN were followed for more than five years with cytology, colposcopy and Vira Pap. Coexistence of HPV infection was sought using a simplified HPV detection kit, the Vira Pap method (Dot Blot hybridization)., Results: Regressive disease was noted in 21 cases, and persistent and progressive disease was noted in 22 cases. HPV DNA was negative in 81% (17 of 21) of regressive disease and positive in 55% (12 of 22) persistent and progressive disease. LCIN had disappeared in 17 (63%) of 27 cases negative for HPV DNA and was persistent or progressive in 12 (75%) of 16 cases positive for HPV DNA., Conclusion: The clinical course of LCIN correlates well with HPV infection.

Cadmium induces the expression of the 70 kDa heat shock protein (HSP70) and metallothionein (MT), both of which are considered to be associated with intracellular glutathione (GSH) metabolism in the cellular protection mechanism against cadmium-induced cellular injury. We determined the effects of N-acetyl-L-cysteine (NAC), which increases the intracellular GSH levels, on the induction of HSP70 and MT gene expression in a cultured cell line of human amniotic cells (WISH) exposed to CdCl2. The mRNA level of MT-II, a major isoform of MT genes, was more prominently increased than that of HSP70 when WISH cells were exposed to CdCl2 (5-15 microM, for 6 h). The treatment of WISH cells with 1.5 and 30 mM NAC for 2 h increased the intracellular GSH levels by 1.4- and 3.1-fold, respectively. Pretreatment of cells with 30 mM NAC significantly reduced both HSP70 and MT-II mRNA levels in the cells exposed to 50 microM CdCl2. This concentration of NAC also efficiently suppressed the cadmium-induced lethality. On the contrary, pretreatment with 1.5 mM NAC suppressed only the induction of HSP70 gene expression in the 50 microM CdCl2-treated cells, and did not inhibit the metal toxicity. However, this low concentration of NAC efficiently suppressed lipid peroxidation which was increased by 50 microM CdCl2. Furthermore, this low concentration of NAC also decreased the CdCl2-induced gene expression of HSP32 which represents a general response to oxidative stress. Taken together, NAC seems to have at least two concentration-dependent functions in WISH cells exposed to CdCl2; the low concentration of NAC can suppress the induction of HSP70 gene expression as well as the increase of lipid peroxidation via an antioxidant pathway, while the high concentration of NAC can suppress the induction of MT-II mRNA as well as cadmium-induced cell death. Our present data suggest that changes in intracellular redox status, as reflected by GSH concentration, have more important effects on the induction of HSP70 mRNA rather than that of MT-II mRNA in human amniotic cells exposed to cadmium.

Objective: The purpose of this study was to determine the incidence of positive peritoneal cytology and to elucidate the prognostic value of peritoneal cytology in patients with uterine cervix and endometrial cancer., Materials and Methods: The incidence of positive peritoneal cytology was investigated in 642 patients including 339 uterine cervix and 303 endometrial cancers. Survival was estimated by the Kaplan-Meier method in a subgroup of 116 stage II cervix and 199 stage I endometrial cancers, and multivariate analysis using Cox's proportional hazards model was used to identify an independent prognostic factor., Results: The incidence of positive peritoneal cytology was found to be 9% in uterine cervix cancer and 15% in endometrial cancer. The incidence was higher in patients with some clinicopathologic status such as advanced stage, lymph node metastasis, ovarian metastasis, and deeper myometrial invasion. The 5-year survival rate for patients with positive or negative peritoneal cytology was 44 or 80% in stage II cervix cancers and 80 or 92% in clinical stage I endometrial cancers, respectively. Multivariate analysis revealed that independent prognostic determinants were pelvic and paraaortic lymph node metastasis and peritoneal cytology in stage II cervix cancer and peritoneal cytology in stage I endometrial cancer. Proper treatment protocol should be scheduled for patients with positive peritoneal cytology.

Few investigations have been made concerning hormonal changes and dyspareunia in fertile aged women with alcoholics experiencing sexual dysfunction. Twenty-seven Japanese woman with alcoholics under 40 years of age excluded with liver cirrhosis were studied to describe alcohol drinking related to sexual dysfunction. Among 21 sexually active women, 20(95.2%) had both symptoms of dyspareunia and vaginal dryness, and only one had neither symptom. Most of patients have lower estradiol levels and 92.0% of patients have the moderately elevated prolactin levels. Eleven of them were having the second grade amenorrhea associated with hyperprolactinemia and hypergonadotropic hypogonadism and 14 were having the first grade amenorrhea. In this study alcoholic abuse women may have deeply related to the hyperprolactinemia, dyspareunia, amenorrhoea, vaginal dryness, ovarian dysfunction and fetal alcohol syndrome.

Ten young adults (15-22 years of age) with acute lymphoblastic leukemia were treated with the protocol of internal medicine and pediatrics co-operative study group since 1989. All patients had complete remission within 5 weeks. Four of 6 patients of low risk group (less than 30,000/microliter of WBC count at diagnosis) and 1 of 4 patients of high risk group have continued the complete remission. Three patients relapsed in bone marrow (BM), and 2 patients in both BM and central nervous system (CNS). All patients could be treated without serious complications except for infection. We obtained the good results of treatment for the low risk group, but did not for the high risk group in this study. Little is known about acute lymphoblastic leukemia in young adults because these individuals are at the borderline age between internal medicine and pediatrics. Taken together with the psychologically distinct age of these patients, it is necessary to establish a closer relationship between internal medicine and pediatrics.

We describe 3 patients with acute myeloblastic leukemia (AML), who received rhG-CSF for infections such as pneumonia or for prophylaxis of infection, and who achieved complete remission. They had not received any antileukemic therapy before or during the administration of rhG-CSF. These findings suggest the possibility that complete remission can be brought about by G-CSF itself in some patients with AML.

Objective: Endometrial cytology in nine cases of ectopic pregnancy was examined in order to elucidate the cellular characteristics of the Arias-Stella reaction and decidual change., Study Design: The cellular findings of epithelial and stromal cells were compared with histologic findings in each case. Furthermore, the immunocytochemical reactivity of each type of cell cluster-epithelial cells without atypia, epithelial cells with atypia, deep stromal cells and sheetlike stromal cells-to epithelial membrane antigen (EMA) or vimentin was investigated and compared with cytologic findings in smears stained with Papanicolaou stain., Results: The distribution of each type of cell cluster correlated fairly well with the histologic findings on the endometrium. Immunocytochemical examination revealed that EMA expression coincided with cell origin identified by Papanicolaou stain. Epithelial cells with atypia, probably corresponding to the Arias-Stella reaction, frequently showed positive reactivity to antivimentin antibody as well as anti-EMA antibody. Endometrial stromal cells usually indicated negative reactivity to anti-EMA antibody. Deep stromal cells expressed vimentin, but sheetlike stromal cells, thought to be decidual cells, infrequently expressed vimentin., Conclusion: The Arias-Stella reaction may be the result of the regenerating and proliferating activity of endometrial glands. It is still controversial whether sheet-like stromal cells are identical to decidual cells.

Objective: To investigate the cytologic and pathologic features of endocervical lesions in cases of gastric adenocarcinoma metastatic to the uterine cervix., Study Design: From 1986 to 1994, four patients with gastric adenocarcinoma metastatic to the uterine cervix were treated at our department. The cervical cytologic samples were obtained by swabbing and were stained by the Papanicolaou method. Presence of tumor diathesis, number of atypical cells, cell arrangement, cytoplastic vacuoles, cellular and nuclear diameter, chromatin distribution and size of the nucleolus were investigated., Results: The smear backgrounds were dirty (tumor diathesis) in two cases and clean in two. No significant difference in the number of atypical cells or in cell or nuclear diameter between primary and metastatic adenocarcinoma was shown. Cell arrangement was the different cytologic finding between primary and metastatic adenocarcinoma. Sheetlike or isolated arrangement was seen frequently in metastatic cervical adenocarcinoma., Conclusion: Because different cytologic features have been found in past and the present series, cytologic diagnosis of metastatic cervical adenocarcinoma should be made carefully.

Pregnancy in a woman with active acromegaly is very rare, because amenorrhea, due to hyperprolactinemia and disturbed pituitary gonadotropin secretion may cause infertility. We report a 28-year-old pregnant woman with untreated acromegaly, who was followed up from early pregnancy to delivery. Her pregnancy was uneventful, and she went into spontaneous labor at 38 weeks and delivered a normal infant. Her serum GH levels were further increased in late pregnancy, followed by decreased in postpartum periods, which may be associated with enlargement of pituitary adenoma during pregnancy. In contrast with serum GH, her serum insulin-like growth factor-1 (IGF-1) levels were dissociated with her serum GH levels during late pregnant and postpartum period. Her serum GH and IGF-1 levels in late pregnancy were different from the levels in pregnant women with acromegaly reported previously.

Adriamycin (ADR) has been considered to target mainly DNA metabolism in the nucleus. Recently, we observed the nuclear translocation of heat shock protein 70 (HSP70) after ADR treatment. We examined which intranuclear changes might be related to this alteration of HSP70 localization. We found considerable alternations in the nucleolar morphology and function in ADR-treated tumor cells, i.e., a ring-shaped segregation of granular components of almost all nucleoli and a dramatic reduction of nucleolar 45S ribosomal precursor RNA biosynthesis in HeLa cells exposed to 100 microM ADR for 2 h. Concomitantly with these changes, HSP70 was concentrated into the nucleoli, as in the case of heat shock treatment. These results indicate a novel anticancer effect of ADR via the suppression of cellular protein biosynthesis, in addition to its effect on DNA.

Objective: To investigate the accuracy rates of cytology, colposcopy and punch biopsy in early cervical neoplasia confirmed by conization., Study Design: During the 10 years from 1984 to 1993, cold knife conization was performed on 151 patients with early cervical neoplasia proven by punch biopsy at our department. The accuracy rates of cytology, colposcopy and punch biopsy were investigated., Results: The accuracy rates of cytology, colposcopy and punch biopsy were 52% (78 of 151), 66% (100 of 151) and 66% (100 of 151), respectively., Conclusion: These results suggest that a composite diagnosis with cytology, colposcopy and punch biopsy is necessary for a correct evaluation. Early cervical neoplasms are frequently seen in young women, and conservative procedures, such as conization, cryosurgery and laser vaporization, are the treatments of choice in order to preserve reproductive function. We recommend conization as the best conservative procedure, with preservation of reproductive function.

We describe a patient who has a chronic polyclonal B lymphocyte proliferation with a hairy-cell appearance. A 48-year-old Japanese woman with marked splenomegaly, systemic lymphadenopathy, and leukocytosis was referred to out hospital. Laboratory examination revealed marked polyclonal IgG hypergammaglobulinemia. Morphologic examination of the patient's peripheral blood, including May-Giemsa staining and scanning electron microscopy, showed a monotonous proliferation of hairy-appearing mature lymphocytes. An immunophenotypic study revealed an expansion of cells with mature B cell antigens positive for CD11c; however, light-chain restriction was not seen. The lack of both immuno-globulin heavy-chain and T cell receptor gene rearrangements by Southern blot analysis indicated the polyclonal nature of the proliferating B cells. This was confirmed further by a clonal analysis of the patient's lymphocytes using an X-chromosome-linked restriction fragment polymorphism within the X-linked phosphoglycerate kinase (PGK) gene. Since chronic B cell lymphoproliferation with a hairy cell appearance has not been described previously, this case might be extremely rare, and has important implication for the pathogenesis of mature B cell lymphoproliferative diseases, including hairy cell leukemia.

The remarkable contraction of small arteries occurred in the perimetrium of the uteri with leiomyomas by a presurgical medication P6th the luteinizing hormone-releasing hormone (LHRH) agonist (buserelin). The contracted vessels showed a significant increase in number of the Weibel-Palade (WP) bodies which occasionally underwent the degranulation and the discharge in a manner of exocytosis. By immunoelectron microscopy, localizations of endothelin (ET)-1 immunoreactive gold particles on the WP bodies as well as on cisterns of the rough endoplasmic reticulum (rER) and Golgi apparatus were pronounced in the buserelin-medicated group when compared to the non-medicated one. These findings indicate that the increase of the WP bodies storing ET-1, of which synthesis in the rER-Golgi system enhances, predominantly occurred in the small arteries in the perimetrium by the buserelin-medication prior to the hysterectomy. ET-1, released into the subendothelial layer, seems to play an important role in the contraction of the vascular media in a manner of paracrine.

A 62-year-old man with aleukemic Bence Jones type multiple myeloma who developed neurologic abnormalities is reported. After admission, consciousness disturbance appeared and a lumbar puncture obtained M-protein. Though brain CT showed no abnormal findings except a punched out lesion of the temporal bone, MRI disclosed remarkable enhancement of the dura mater. Meningeal involvement by myeloma cells without leukemic blood picture is very rare though it is common in other lymphoproliferative disorders such as acute lymphocyte leukemia and malignant lymphoma. We report a case of BJ type multiple myeloma with meningeal involvement due to diffuse infiltration to the dura mater and discuss a possible mechanism of meningeal involvement in this patient.

Oxygen radicals have been implicated as the causative agent in deleterious processes such as gene mutation, cellular transformation and cell death. Cellular response to DNA damage induced by hydrogen peroxide was examined in human amniotic cultured cells (WISH). The content of 8-hydroxydeoxyguanosine in nuclear DNA increased during the early period of exposure to hydrogen peroxide, that is, it peaked at 1 h in the cells treated with 1 mM hydrogen peroxide and then declined, but not to the control levels within 4 h. Interestingly, immunofluorescence staining of treated cells revealed that cytoplasmic hsp70 protein translocated transiently into the nucleus, which was most remarkable around 30 min after the addition of 1 mM hydrogen peroxide into the culture medium. On the other hand, the induction of hsp70 gene mRNA was not significant during the early period of treatment by 0.5 mM hydrogen peroxide and was occurred after 5 h. DNA damage induced by Adriamycin further supported the possible correlation between hydroxyl radical-mediated cleavage of DNA and nuclear translocation of hsp70 protein. These results suggest that preexisting hsp70 protein translocated into the nucleus to either protect chromatin DNA from further damage or to facilitate the repair of DNA damage in some unknown ways.

The induction mechanism of heat shock 70 (Hsp70) gene by cadmium was investigated. In human amniotic WISH cells, Hsp 70 was induced by cadmium in a dose- and time-dependent manner. Cadmium-induced Hsp70 mRNA levels were enhanced 3- to 4-fold after depletion of intracellular glutathione (GSH) by either diethylmaleate or buthionine sulfoximine. Under these conditions, hydrogen peroxide might increase in the absence of substrate for glutathione peroxidase. We found that exogenous hydrogen peroxide alone induced Hsp70 which was further enhanced significantly after GSH-depletion by diethylmaleate. On the other hand, treatment of cells by diethyldithiocarbamate, an inhibitor of superoxide dismutase, induced Hsp70 2-fold over the level of control. This induction was further stimulated by cadmium even in the presence of GSH. Furthermore, a 4-fold increase of intracellular GSH by the treatment of cells with glutathione isopropyl ester did not diminish the cadmium-induced Hsp70. Gel mobility shift assays of nuclear extracts, from these differently treated cells, with oligonucleotide containing a promoter region of Hsp70 gene revealed that the levels of Hsp70 mRNA observed in the present study corresponded to the changes of transcription. These results imply that the induction of Hsp70 mRNA by cadmium is mediated at least partly via reactive oxygen species and attenuated by cellular GSH and that some part of cadmium-induced Hsp70 can not be eliminated by GSH, suggesting that multiple signals are functioning for this induction.

A 26-year-old man was admitted to our hospital with cervical tumor and facial edema on July 8, 1991. Examination of chest X-ray and chest CT showed a bulky tumor in the mediastinum and pleural effusion. A pathological diagnosis of non-Hodgkin's lymphoma (diffuse large cell, immunoblastic type) was made by biopsy of the cervical lymph node. MACOP-B chemotherapy or other combination chemotherapy did not achieve complete remission. The man was given a preparative regimen consisting of busulfan at 16 mg/kg orally and 60 mg/kg of etoposide (Bu-Et); 30 mg/kg of etoposide was administered by continuous intravenous infusion for 12 hours on day-5 and day-4, before he received autologous bone marrow on February 20. He was then given 300 micrograms of G-CSF was given to him to accelerate recovery of hematopoiesis from one day after BMT. The neutrophil count to 500/microliters recovered on day 28, and residual tumors disappeared. Although moderate-grade stomatitis and nasal bleeding developed, these toxicities were controllable and no veno-occlusive disease resulted. Regimen-related toxicities of Bu-Et preparatory regimen have been generally considered to be severe, but continuous and separate administration of etoposide as reported in this case may be useful to reduce side effects of this preparatory regimen.

In order to develop new-generation macrolide antibiotics active against erythromycin (EM)-resistant strains, a series of 6-O-methyl EM 9-O-substituted oxime derivatives was synthesized and evaluated for antibacterial activity against EM-resistant (S. aureus J-109) and susceptible (S. aureus 209P) strains. To understand how substituents affect the biological activity, the quantitative structure-activity relationships (QSAR) was analyzed using the Hansch-Fujita method. With the EM-resistant strain, the positive coefficient for log P may indicate that higher hydrophobicity of molecules is favorable for antibacterial activity. The negative coefficients of the Sterimol parameters L, B1, and B5 may indicate that long, bulky substituents are unfavorable. With the EM-susceptible strain, the negative coefficient for log P may indicate that hydrophilicity is important for antibacterial activity. A short substituent is also required to improve the activity. Based on the QSAR model, a derivative (87) having an anthracenylmethyl moiety was synthesized to reinforce and confirm the correlation. The activity of 87 against the EM-resistant strain was significant. In QSARs of 6-O-methyl EM-A 9-O-substituted oxime derivatives, the difference of the contribution of log P to the antibacterial activity between EM-resistant and susceptible strains was clearly recognized.

A 56-year-old woman was admitted with pyrexia, cough, and dyspnea on August 21, 1991. Physical examination revealed anemia in the palpebral conjunctivas and moist rales at the right lower lung field. Neither the Liver nor spleen was enlarged. Examination of the peripheral blood showed a hemoglobin level of 8.1 g/dl, a platelet count of 14.8 x 10(4)/microliters, and a white blood cell count of 2,800/microliters, with 7% blasts and 8% megakaryocytes. Tear drop-like erythrocytes, agranular neutrophils, and erythroblasts were also seen in the peripheral blood. Examination of the bone marrow showed 15% peroxidase positive blasts, and many micromegakaryocytes. Cytogenetic studies for bone marrow cells revealed the existence of the Philadelphia (Ph1) chromosome. Bone marrow biopsy showed normal cellularity with increase of megakaryocytes and advanced myelofibrosis. Breakpoint cluster region (bcr) rearrangement analysis using the peripheral blood mononuclear cells revealed M-bcr rearrangement. According to the Hannover classification for myeloproliferative disease, she was diagnosed as having CML with advanced myelofibrosis followed by CML with megakaryocytic increase. Since she had neutrocytopenia and severe infectious disease, she received a subcutaneous injection of 125 micrograms of G-CSF. Not only increase of the white blood cell count, but also disappearance of blasts, improvement of anemia, increase of the platelet count, and improvement of myelofibrosis were observed.

Normal human diploid fibroblasts (HDF) have a finite life span in vitro and have been used as a model system for the study of in vivo aging. Little is known about how changes in gene expression may affect the immortalization of human fibroblasts. We looked for cDNA clones whose mRNAs were differentially expressed between mortal senescent SV40-transformed human fibroblasts (B-32) and the immortal counterparts (B-32F) derived from B-32 cells. We identified three cDNA isolates by subtractive differential hybridization with 32P-labeled cDNA probes from B-32 cells and B-32F cells. Nucleotide sequence analysis of these cDNA clones revealed that they were homologous to the human vimentin, a human mitochondrial gene and a human gene of unknown nature. Slot blot and Northern blot analyses demonstrated that the former two were preferentially expressed in senescent B-32 cells and the last one was less expressed in B-32F immortal cells.

Cytologic findings on various vulvar squamous lesions are described in order to elucidate the usefulness of vulvar cytology. Lichen sclerosus, hyperplastic dystrophy and dysplasia with a few exfoliated anucleate squamous cells could not be differentiated cytologically. Numerous parakeratotic cells and dyskaryotic cells were identified in squamous cell carcinoma in situ. Three cytologic patterns were presented in cases of frankly invasive squamous cell carcinoma: negative cytology with parakeratotic cells, suspicious cytology with dyskaryotic cells and positive cytology with malignant cells. Verrucous carcinoma yielded only anucleate squamous cells. Parakeratotic cells without nuclear atypia seemed to be neoplastic cells on vulvar cytology.

The immunocytochemical localization of hCG in the syncytiotrophoblast of hydatidiform moles was observed by means of the immunoglobulin gold technique with rabbit anti-human hCG serum. Immunoreactive gold particles of hCG are preferentially localized in the distal portion of the Golgi complexes, Golgi-derived secretory granules 200-300 nm in diameter, and large electron dense bodies 500-1,000 nm in diameter. The secretory granules exist throughout the cytoplasm above the nuclear region and are occasionally released by exocytosis from the syncytiotrophoblast. On the other hand, the large electron dense granules seem to be cytolysosomes in nature and are involved in uptake and storage of hCG.

Although erythromycin A contains five hydroxyl groups, regioselective methylation at the C-6 hydroxyl group was achieved to the extent of 90% when a 9-O-substituted erythromycin A 9-oxime was employed as substrate. The methylation and its selectivity are dependent on an O-protecting group at the 9-oxime, solvent, base, and methylating reagent. In particular, the use of a polar aprotic solvent is indispensable for the methylation. Among the 9-oxime derivatives, 2'-O,3'-N-bis(benzyloxycarbonyl)-N-demethylerythromycin A 9-[O-(2-chlorobenzyl)oxime] was the most important intermediate for the synthesis of clarithromycin (6-O-methylerythromycin A).

There are several reasons why Papanicolaou classification should not be used for the interpretation of cytologic findings. First, many different classification criteria are sometimes used for a single pathologic condition. Second, inflammatory change or hormonal evaluation are not included in the system. Third, the classification is not compatible for the evaluation of endometrial lesions or chorionic diseases. Fourth, mismanagement of the patient is sometimes encountered when this classification is only used. Instead of Papanicolaou classification, descriptive diagnosis should be used for cytologic interpretation.

Magnetic Resonance Imaging (MRI) has been widely used for the diagnosis of gynecologic disease. We studied the clinical usefulness of MRI in 34 patients with endometrial cancer from July, 1988 to August, 1991. Pre-operative pelvic MRI was carried out to evaluate the myometrial invasion in these patients. MRI was accurate in predicting the presence or absence of myometrial invasion in 23 of 34 cases (68%), while echo or CT (computed tomography) was accurate in 26% and 8%. In conclusion, MRI is considered to be the most useful examination for the evaluation of myometrial invasion in endometrial cancer.

We describe siblings with the non-salt-losing form of 21-hydroxylase deficiency who had hypersecretion of aldosterone and plasma renin activity (PRA). Blood pressure and serum electrolytes in both cases were normal despite the aldosterone hypersecretion. Aldosterone secretion was elevated markedly with ACTH administration and with sodium deprivation and/or volume depletion during ACTH suppression by dexamethasone. With suppression by dexamethasone, aldosterone hypersecretion was decreased with lowering of the steroids proximal to the block in the biosynthetic pathway. However, urinary sodium excretion was decreased. These results suggest that the biosynthetic pathway for aldosterone production was preserved. Furthermore, aldosterone hypersecretion and high PRA may serve to compensate for the sodium loss which results in turn from the overproduction of the sodium-losing steroids, such as progesterone and 17 alpha-hydroxyprogesterone which are aldosterone antagonists.

The usefulness of MR imaging (MRI) in uterine cervical cancer after radiotherapy was evaluated in 18 patients. MRI was performed before and within one month after radiotherapy. In eight patients the uterus assumed a nearly normal appearance with resolution of the tumor on MRI. In another eight patients the tumors resolved, but an area of high signal intensity remained at the cervix. In these patients the tumors were well controlled at clinical examinations and during the mean follow-up period of 15-18 months. Therefore a persistent area of high signal intensity alone at the cervix on MRI is not itself indicative of residual tumor. An area of high signal intensity and a residual cervical mass persisted in two cases, and they were histologically proven to be due to residual tumor. Both patients died of recurrent disease at seven and 11 months after their radiotherapy. We conclude that MRI is useful for evaluating patients with cervical cancer at the completion of radiotherapy, and that MRI complements conventional clinical examinations.

Combination chemotherapy with Cyclophosphamide, Adriamycin and Cisplatin (CAP) is one of the most effective chemotherapies for ovarian cancer. Severe vomiting induced by this treatment may cause a disorder of the serum electrolytes. Severe hyponatremia was observed in one patient who complained of convulsions and unconsciousness. Accordingly we studied the disorder of serum electrolytes in 158 courses of CAP treatment (44 patients) during the five years from 1984 to 1988. The serum electrolytes were influenced by the amount of vomiting in this study. Consequently frequent examination of serum electrolytes is necessary during this treatment.

To elucidate the accuracy of cytology in diagnosing cervical intraepithelial neoplasia (CIN) during pregnancy, cytologic screenings for uterine cervical cancer in pregnant women were reviewed for a five-year period. Of the 967 pregnant women screened, abnormal cytologic findings were recorded for 15 (1.6%). Only nine of these were subsequently examined by colposcopy and punch biopsy, which demonstrated CIN in all cases, for an incidence of documented CIN during pregnancy of 0.93%. Including two referral cases also examined by colposcopy and biopsy, cytology and histology agreed on the degree of CIN in four cases and disagreed by one degree in four cases, by two degrees in two cases and by three degrees (mild dysplasia versus carcinoma in situ) in one case. Review of the specimens from these cases did not readily explain the poor concordance between cytology and punch biopsy; some findings suggest that overestimation of the punch biopsy sample may be the explanation.

In the management of a chorionic disease, human chorionic gonadotropin (hCG) is the most reliable tumor marker. However, hCG has a low titer, so that it cannot be strictly distinguished from the human luteinizing hormone (hLH) by the traditional method that uses a hemagglutinin reaction (HAR) or by a radioimmunoassay (RIA). Recently, however, the detection of the beta-COOH-terminal peptide of hCG (hCG-CTP) by an enzyme immunoassay has made it possible to clearly distinguish hCG from hLH, and from April, 1987 to December, 1989, 13 trophoblastic diseases have been managed using this new technique. Results have shown that human chorionic gonadotropin-CTP, when compared to other methods of measurement, is the most sensitive tumor marker and as it is the most accurate, it should be used in careful follow-up observations of a chorionic disease.

A rare case of placentae site trophoblastic tumor (PSTT) studied by immunohistochemistry and nuclear DNA analysis is reported. The patient, a 24-year-old Japanese female, complained of amenorrhea. Dilatation and curettage revealed a small specimen that contained trophoblastic cells and caused intractable bleeding. Pelvic sonography revealed a 5-cm mass in the posterior uterine wall with multiple cystic lesions of several sizes. The cystic lesions were shown to be dilated vessels by magnetic resonance imaging (MRI) and digital subtraction angiography (DSA). Serum beta-hCG (beta subunit of human chorionic gonadotropin) was 3.7 ng/ml. Total abdominal hysterectomy revealed a well-circumscribed, yellow, soft mass in the posterior uterine wall. Microscopic findings were consistent with PSTT and the mitotic count was extremely low. Immunohistochemically, most of the tumor cells were intensely stained with human placental lactogen, whereas few were stained with human chorionic gonadotropin. The nuclear DNA content of the trophoblastic cells showed a sharp peak at the triploid range coexistent with a few cells of higher ploidy. This is the first report of sonographic findings and nuclear DNA analysis by spot cytometry in a case of PSTT.

Seventeen multiple primary cancers including 16 double cancers and one triple cancer were found in 316 patients with gynecologic malignancies who were treated in our department from 1984 to 1988. All pathologic slides but one were reviewed, and cases with possible metastasis or recurrence were not included in this study. The incidence of multiple primary cancers in gynecologic malignancies was 5.4%. Multiple primary cancers were encountered in 4.4% of 205 cervical cancers (including carcinoma in situ), 15.2% of 33 endometrial cancers, and 8.6% of 58 ovarian cancers (including low potential malignancy), respectively. The most frequent sites of other cancers were seen in the large intestine and rectum (5/17), breast (4/17), and gynecologic organs (3/17). Higher incidences were seen in our study that in those in domestic literature. This is probably because detailed anamnesis and gastrointestinal series were obtained in most gynecologic malignancies (especially in endometrial or ovarian cancer).

An extremely rare case of in situ adenocarcinoma of the uterine cervix that developed into invasive cancer is reported. The patient was a 54-year-old Japanese female who complained of vaginal spotting. Pelvic examination revealed no abnormalities and routine Papanicolaou smear was misinterpreted as negative. Multiple wedge biopsies of the uterine cervix were done because Papanicolaou smears taken after 7 months were reported as suspicious. The wedge biopsy revealed dysplastic changes of the endocervical glands. It was diagnosed as being in situ adenocarcinoma by a later review. The patient was examined by cytology and colposcopy every 3 to 12 months. An advanced adenocarcinoma was found in the uterine cervix after 5 years. Total abdominal hysterectomy, resection of the vagina and cardinal ligament, and postoperative irradiation were performed. The patient is alive and well at this writing. This is the second case of in situ adenocarcinoma of the uterine cervix that developed into invasive cancer.

Estrogen receptor (ER) content in myoma and myometrium was determined by radio-receptor assay (RRA), and ER staining was performed by immunocytochemical staining, i.e. the peroxidase-antiperoxidase (PAP) method, in uterine samples from eight patients with uterine leiomyoma. ER content in myoma tended to be hither than that in myometrium, but the difference was not significant. The distribution pattern of ER staining was thought to be an important criterion. When the data were compared on the basis of the number of cells in the site where the largest number of positive cells aggregated, the number for myoma was significantly higher than that for myometrium (P less than 0.05). These results suggest that myoma contains more ER positive cells to obtain estrogen effects than myometrium.