Influence of laser power on atom probe tomographic analysis of boron distribution in silicon.

The relationship between the laser power and the three-dimensional distribution of boron (B) in silicon (Si) measured by laser-assisted atom probe tomography (APT) is investigated. The ultraviolet laser employed in this study has a fixed wavelength of 355nm. The measured distributions are almost uniform and homogeneous when using low laser power, while clear B accumulation at the low-index pole of single-crystalline Si and segregation along the grain boundaries in polycrystalline Si are observed when using high laser power (100pJ). These effects are thought to be caused by the surface migration of atoms, which is promoted by high laser power. Therefore, for ensuring a high-fidelity APT measurement of the B distribution in Si, high laser power is not recommended.

Enhancement of tumor growth and metastases by medroxyprogesterone acetate in transplanted uterine adenocarcinoma cells of the rat.

PIP: This study describes the effects of medroxyprogesterone acetate on tumor growth and metastases in transplanted uterine adenocarcinoma cells of the rat. High-and-low-tumorigenic cloned cells of Sprague-Dawley rat uterine adenocarcinoma originally induced by 7,12-dimethylbenz (alpha) anthracene in vivo were used. Both were derived from the same parent culture. They were cultured for more than 2 years and both retained almost the same transplantability. Survival rate of cell colonies in vitro was reduced in both lines after progesterone treatment of more than 8 mcg per ml. This reduction was dose dependent. About 1 million cells suspended in .2 ml culture medium were injected sc into the interscapular region of isologous newborn rats. At 5 weeks these rats were given .5 mg medroxyprogesterone acetate twice a week for 2 weeks. At 7 weeks they were killed. High-tumorigenic cells produced growing tumors in all newborn rats. About a third of these rats died of metastases during the 7-week observation period. Tumors produced by low-tumorigenic cells grew slowly and occasionally regressed without metastases to the lung. Tumors in female rats were larger than those in males. Enhancement of tumor growth and metastases by this progesterone compound was observed in rats inoculated with low-tumorigenic cells as compared to controls. The enhancement was not significant in tumors produced by high-tumorigenic cells. The progesterone may act immunosuppressively in vivo, or make alterations in environmental conditions of the tumors.^ieng

In vivo and in vitro tests of inhibitory effect of progesterone on cell-mediated immunity in rats bearing a syngeneic uterine adenocarcinoma.

Growth inhibition or stimulation of target adenocarcinoma cells in rats sensitized with spleen cells from syngeneic tumor-bearing rats was significantly suppressed in colony inhibition assays when the spleen cells were treated in vitro with 0.8 mug progesterone or more/ml medium. In addition, when tumor-bearing rats were treated with 1 mg medroxyprogesterone acetate weekly in vivo, the inhibiting action of the spleen cells from rats with regressed tumors was also suppressed. Progesterone thus suppressed immune spleen cells in vivo and in vitro.

Spontaneous occurrence of atypical hyperplasia and adenocarcinoma of the uterus in androgen-sterilized SD rats.

In SD female rats sterilized by a single injection of testosterone propionate at 2 days after birth, the spontaneous occurrence of atypical hyperplasia and adenocarcinoma of the uterus was observed for a fairly long period (greater than 2 yr). Two atypical hyperplasias and 2 adenocarcinomas were detected in 25 androgen-sterilized rats (ASR) after 500 days of age; in contrast, in 111 normal control rats no abnormal uterine proliferation was detected during a 750-day observation period. These results indicate that a persistence of both hormone imbalances and dysfunctional uteri in ASR induces abnormal uterine proliferation at a late age.

Cellular differentiation of rat uterine adenocarcinoma cells by progesterone in vitro.

Transplantable cloned HTP/Cl culture was a stable line derived from a rat uterine adenocarcinoma that was induced by 7,12-dimethylbenz(a)anthracene in vivo and did not display density-dependent inhibition of growth. This HTP/Cl culture easily adapted to grow in a culture medium containing progesterone, 8 mug/ml. As compared with HTP/Cl culture, HTP/Cl/P8 culture grown in the presence of progesterone was contact inhibited, and a cellular differentiation was observed in the tumor tissues that developed after inoculation of cells. These actions of progesterone on uterine adenocarcinoma cells were completely reversible on removal of the hormone in vitro. These results appeared to indicate that progesterone was involved in the regulation of both cellular proliferation and differentiation. The possible mechanisms of the regulation of rat uterine adenocarcinoma cells by progesterone are discussed in relation to cellular levels of cyclic adenosine 3':5'-monophosphate in vitro.

Molecular properties of F9 embryoglycan recognized by a unique antibody in sera from patients with germ cell tumors.

Human antibody against an embryoglycan present on a mouse teratocarcinoma cell line F9 was found in sera from 16 of 29 patients with embryonal carcinoma, yolk sac tumor, immature teratoma, and choriocarcinoma of gonadal and extragonadal origins by Farr assay. In contrast, none of the sera from patients (77 cases) with dysgerminoma, seminoma, germinoma, and mature teratoma or from patients (118 cases) with nongerm cell types of ovarian tumors contained this antibody. The antigenic embryoglycan was of high molecular weight (Mr greater than 70,000) on Sephacryl S300 column chromatography and carried binding sites for Grifonia simplicifolia agglutinin-1. The antigenic embryoglycan was also found in F9 cell-cultured medium. Absorption of patients' sera with synthetic Blood Group B trisaccharides failed to remove the antibody against F9 embryoglycan. None of these patients' sera showed higher hemagglutination titer to rabbit erythrocytes than the normal range. In contrast, alpha-galactosyl carbohydrates obtained from Ehrlich ascites tumor cells effectively inhibited the binding of patients' sera with F9 embryoglycan. These results indicate that the human antibody against F9 embryoglycan recognizes alpha-galactosyl structures that are distinct from B blood group antigen, but are cross-reactive with alpha-galactosyl structures on Ehrlich ascites cells.

Neural rosette formation within in vitro spheroids of a clonal human teratocarcinoma cell line, PA-1/NR: role of extracellular matrix components in the morphogenesis.

From the human teratocarcinoma-derived cell line PA-1, we established a clonal line, PA-1/NR, that stably produced a distinct cellular arrangement of neural rosettes when cultured as in vitro multicellular spheroids for 3 weeks. On immunofluorescence staining and fluorescence-activated cell sorter analyses, PA-1/NR cells in monolayer expressed the neuroectoderm-associated antigens HNK-1, NC-1, and A2B5 and the neuroblastoma-associated antigens KP-NAC8 and KP-NAC10 but lacked human embryonal carcinoma antigens, SSEA-3 or K21 antigen. Here, we investigated the developmental process of rosette formation with respect to morphological features, distribution of mitotic cells, and expression of multiple lineage-related markers and extracellular matrix (ECM) components. Ultrastructural examination of these rosettes disclosed a well-defined cavity radially surrounded by wedge-shaped or pseudostratified cells, apical microvilli and junctional complexes, and basal laminae and collagen fibrils at their basal surface. In these rosettes, many proliferating cells were detected by the immunohistochemical staining of cells incorporating bromodeoxyuridine. PA-1/NR spheroids consistently displayed neuron-specific enolase, S-100 protein, and vimentin but not glial fibrillary acidic protein, neurofilament proteins, or myelin basic protein. The rosette formation accompanied a strikingly polarized and overlapped deposition of ECM components including tenascin-carrying HNK-1 epitopes, laminin, type IV collagen, heparan, and chondroitin sulfate proteoglycans. Immunoblotting analyses showed that laminin B1 and B2 chains were constitutively expressed, whereas a fully assembled form of laminin and type IV collagen appeared only after spheroid development, suggesting that these ECM components play a morphogenetically important role in rosette formation. Close similarities between these rosettes and the neural tube of humans and experimental animals in the morphogenetic process and ECM formation lead us to propose that the PA-1/NR spheroids provide an in vitro model for the study of the earliest stage of human neurogenesis.

In vivo and in vitro studies of experimental ovarian adenocarcinoma in rats.

When 7,12-dimethylbenz[a]anthracene-impregnated sutures were directly applied to the ovarian parenchyma of 8-week-old Sprague-Dawley rats (the clipping method), adenocarcinomas developed in 29 (39%) of the 75 rats during the 50-week observation period. When 20-methylcholanthrene was used, adenocarcinomas developed only in 1 (3%) of the 31 rats. Thus, the clipping method using 7,12-dimethylbenz[a]anthracene is satisfactory as an animal model of ovarian adenocarcinoma which comprises 85 to 90% of human malignant ovarian tumors. On the other hand, attempts were made to isolate cloned cell lines from these experimental ovarian adenocarcinomas in vitro, and two cloned cell lines were obtained. They were epithelioid and produced undifferentiated adenocarcinomas by back-transplantation into isologous newborn rats.

CA54/61 as a marker for epithelial ovarian cancer.

Using a new one-step, double-determinant enzyme immunoassay, we performed quantitative measurements of a mucin-type glycoprotein antigen (CA54/61) that we recently detected in sera of ovarian carcinoma patients. When the cutoff value was set at 12 units/ml, at which a high diagnostic efficiency was demonstrated [or at 20 units/ml (mean + 3 SD of healthy females)], the positive rates of ovarian serous, mucinous, clear cell, and endometrioid carcinomas were 76% (or 63%), 63% (or 55%), 57% (or 52%), and 50% (or 38%), respectively. Even in mucinous cystadenocarcinoma, more than one-half of the cases were positive, indicating the potential utility of the assay in the diagnosis of mucinous tumors. In sera from patients with benign ovarian tumors, only 9% (or 4%) of the cases were positive, indicating the quite high specificity of this test for ovarian carcinomas. To make a comparison between CA54/61 and CA125, we set the cutoff level of CA125 at 110 units/ml, at which value a high diagnostic efficiency was demonstrated [or at 35 units/ml (mean + 3 SD of healthy females)]. When both CA54/61 and CA125 were assessed in sera from 36 patients with mucinous cystadenocarcinoma, the positive rates of CA54/61 and CA125 were 64% (or 56%) and 36% (or 56%), respectively, suggesting that CA54/61 is of clinical value as a new tumor marker for ovarian cancers, including mucinous tumors.

Poly-N-acetyllactosamines synthesized by cultured Ehrlich carcinoma cells: application of endo-beta-galactosidase C for analysis of the terminal structure.

Poly-N-acetyllactosamines were prepared from Ehrlich carcinoma cells cultured in the presence of [14C]galactose. Methylation analysis indicated that 31% of the galactose was in the non-reducing end. Of it, 77% was cleaved by alpha-galactosidase, and 56% was released as a disaccharide by endo-beta-galactosidase C. Methylation analysis confirmed that the released disaccharide was mostly Gal alpha 1----3Gal. Therefore, Gal alpha 1----3Gal structure, not Gal alpha 1----3(Gal alpha 1----6)Gal structure, was the major alpha-galactosyl structure in the poly-N-acetyllactosamines synthesized. Furthermore, alpha-galactosidase digestion did not change the content of disubstituted galactosyl residues. Thus, Gal alpha 1----3(Gal alpha 1----6)Gal structure, which was suggested to be the sole non-reducing terminal structure of poly-N-acetyllactosamines of Ehrlich carcinoma cells, was not detected in significant amounts under the present experimental conditions.

Serum placental alkaline phosphatase (PLAP) in gynecologic malignancies--with special reference to the combination of PLAP and CA54/61 assay.

Serum placental alkaline phosphatase (PLAP) levels in patients with gynecological tumors were measured by two kinds of enzyme-antigen immunoassay kits provided by Innogenetics (PLAP-I) and Sangtec Medical (PLAP-S), and the combination assays for PLAP with other tumor markers were studied. None of the healthy women studied were PLAP positive, and the positive rate in patients with benign ovarian tumors was less than 6%. The positive rate in patients with ovarian cancers was about 35%, which was higher than the rates for other cancers. It was significantly higher in patients with ovarian serous cystadenocarcinoma (60%). Remarkably high PLAP-I values were observed in patients with dysgerminoma. By the combination assay for PLAP with CA54/61 antigen in ovarian cancers, the diagnostic efficiency increased compared with that for PLAP and CA125. We conclude that PLAP is useful in the diagnosis of ovarian serous cystadenocarcinoma and dysgerminoma and that CA54/61 is the better partner for the combination assay.

Human cell clones, RSa and UVr-1, differing in their capability for UV-induced virus reactivation and phenotypic mutation.

UVr-1 is a human cell clone established as a variant with increased resistance to cell killing by ultraviolet light (UV, principally 254 nm wavelength) from a UV-sensitive cell clone, RSa. Both cells have been characterized to have much the same capacity of UV-induced DNA repair synthesis in whole cells, and the parent RSa cells were recently found to be hypermutable. In the present study UVr-1 cells were characterized in comparison RSa cells with respect to UV-induced virus reactivation and phenotypic mutation. Survival levels of UV-irradiated vaccinia virus and herpes simplex virus type 1 (HSV-1) were much the same in logarithmically proliferating UVr-1 and RSa cells. Correlated with these host cell reactivation levels, the same extent of UV-induced DNA repair replication synthesis was observed in isolated nuclei of the two cell clones. Enhancement of survival levels of UV-irradiated HSV-1 was detected when proliferating RSa cells were irradiated with UV prior to the virus infection. In contrast, this enhanced virus reactivation (EVR) was not detected in similarly irradiated and infected UVr-1 cells. As for phenotypic mutation frequencies assessed by the cloning efficiency of cells with increased resistance to ouabain cell killing (OuaR), OuaR mutants were not obtained from UVr-1 cells either with or without UV irradiation. When the proliferation of cells was synchronized, both EVR and OuaR mutations were detected in RSa cells irradiated with UV at any cell cycle phase, being greatest in the later half of the G1 phase. However, there was no detectable EVR or mutation in any phase of synchronous UVr-1 cells. The hypomutability of UVr-1 cells and hypermutability of RSa cells in a G1 cell cycle phase was also found even if 4-nitroquinoline 1-oxide was used as a mutagen or mutant cells with increased resistance to 6-thioguanine cell killing were estimated.

Glycopeptides from murine teratocarcinoma cells: structure of the determinants recognised by Griffonia simplicifolia agglutinin I and by sera from patients with ovarian germ cell tumors.

High-molecular-weight glycopeptides synthesised by teratocarcinoma OTT6050 bear the binding site for Griffonia simplicifolia agglutinin I and are recognised by antibodies in the sera of patients with ovarian germ cell tumors. Digestion of the glycopeptides with endo-beta-D-galactosidase C abolished the lectin binding activity and the antigenic activity. Since the product of the enzymic digestion is alpha-D-Gal-(1----3)-D-Gal, it is concluded that the disaccharide structure is involved in the lectin binding site and the antigenic site.

More confined indications for use of combined passive and active immunization for preventing perinatal development of hepatitis B virus carrier-state--based on the natural history of hepatitis B virus-vertical transmission.

To find more confined criteria for use of passive and/or active immunization for preventing perinatal development of hepatitis B virus (HBV) carrier-state than maternal HBe antigenemia, maternal HBsAg-titers (R-PHA) around delivery and infantile HBeAg-titers-(EIA) are discussed. No children whose maternal HBsAg-titers around delivery were lower than 3(6) developed carrier-state in spite of maternal HBe antigenemia. In addition, at age 2 months serum HBeAg-titers of 6 children who had acquired persistent HBsAb were lower than 25, while those of 5 children who had developed carrier-state were higher than 70. These findings may contribute to the establishment of more confined indications for the administration of HBIG and/or HB vaccine to the children born to HBeAg-positive carrier women, saving not only HBIG and HB vaccine but all accompanied efforts of both patients and medical staff as well.

Cytocidal effects of hematoporphyrin derivative and argon dye laser on human gynecologic tumor cells in vitro.

Antitumor effects of hematoporphyrin derivative (HpD) plus argon dye laser were examined using various human gynecologic tumor cells in vitro. Irradiation alone with argon dye laser showed no effect on the DNA synthesis of cells. Treatment of the cells with HpD inhibited the DNA synthesis depending on the concentration and the exposure time. Photoradiation by argon dye laser following treatment of the cells with HpD 40 micrograms/ml for 2 h killed more than 80% of cells when the cells were irradiated for more than 3 min. Prominent degenerative changes of the cytoplasm and the nucleus appeared within 1 h after photoradiation. These changes were confirmed by morphology and DNA histogram. There were no differences of sensitivity to photoradiation among the three histologically different kinds of cells lines used.

Reproductive status in GTD treated with etoposide.

OBJECTIVE: To investigate the mechanism and degree of ovarian dysfunction in gestational trophoblastic disease (GTD) patients treated with etoposide alone. STUDY DESIGN: Forty-seven patients with low-risk GTD were treated with etoposide alone, and pituitary-ovarian function was evaluated by measuring basal serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol and progesterone and by recording basal body temperature. Moreover, the responses of LH and FSH to the administration of LH-releasing hormone (LHRH) and the responses of prolactin to thyrotropin-releasing hormone (TRH) were analyzed after the completion of etoposide treatment. RESULTS: Increased basal LH and FSH levels were found in approximately 50% of patients, especially those over 40 years old. Although the LH and FSH responses to LHRH were exaggerated in patients with high basal FSH levels, the prolactin responses to TRH were normal. Ovulation resumed within 121 days after the cessation of chemotherapy in women under 39 years. However, five of nine patients over 40 years remained anovulatory during the follow-up period. CONCLUSION: Ovarian function was impaired in approximately 50% of patients treated with etoposide at the time of LHRH study, though pituitary function was preserved. This complication is age related but not related to the amount of etoposide exposure. Therefore, we must consider the possibility of permanent anovulation when we treat patients 40 years old and older.

Malignant change of dermoid cysts of the ovary. Report on an adenosquamous cell carcinoma and a clear cell carcinoma.

Five cases of malignant change of ovarian dermoid cyst are presented. Two of these cases were of special interest due to the unusual nature of the malignancy. One case (an adenosquamous cell carcinoma) was apparently a malignant change of the columnar epithelium and its squamous metaplasia. The other case (a clear cell carcinoma) appeared to represent coincidental development of two tumors.

Clinical study of recombinant hepatitis B vaccine.

The efficacy and safety of a recombinant yeast-derived hepatitis B vaccine were evaluated in 209 subjects after three administrations at 0, 4 and 20 weeks. Subjects were divided into four groups given 5 micrograms vaccine subcutaneously, 10 micrograms subcutaneously, 10 micrograms intramuscularly and 20 micrograms subcutaneously to define the effective dose and to compare the effect of administration. Seroconversion of the antibody to hepatitis B surface antigen after the third vaccination reached 96.6% in the group given 5 micrograms vaccine subcutaneously and 100% in the other groups. The final geometric mean antibody titres were 700 IU/l in subjects given 5 micrograms subcutaneously, 2004 IU/l in those given 10 micrograms subcutaneously, 4674 IU/l in those given 10 micrograms intramuscularly and 3342 IU/l in those given 20 micrograms subcutaneously. In the groups given 10 micrograms, the early seroconversion rate of the antibody to hepatitis B surface antigen and the geometric mean antibody titres after the third vaccination were significantly higher in subjects administered intramuscularly than subcutaneously (P less than 0.05). No major adverse effects were observed and minor reactions were the same as, or less than, those reported for the plasma-derived vaccine. Before and after administration, no significant fluctuation in the yeast antibody titre was observed. These results demonstrate the efficacy and safety of the yeast-derived vaccine, and show that 10 micrograms was the effective dose.

[Pharmacokinetic and clinical studies of cefuzonam in obstetric and gynecologic infections].

Cefuzonam (CZON, L-105), a new semisynthetic cephem antibiotic, was studied in the field of obstetrics and gynecology, pharmacokinetically and clinically. Following is a summary of the results. 1. Concentrations of CZON in serum and genital organs following intravenous drip infusion of 1 g of the drug over 60 minutes were measured. Samples were taken during 40 to 190 minutes after the end of the infusion. Ratios of concentrations of the drug transferred into genital organs to the concentration in the cubital venous serum were 0.5 to 1.5 in many cases. Levels of the drug in the genital organs examined well exceeded MICs for common causative organisms in obstetric and gynecologic infections. 2. Therapeutic effects of 1 g or 2 g of CZON by drip infusion twice daily were evaluated in 8 patients. Overall clinical efficacy was 62.5% (5/8). No side effects or abnormal laboratory findings due to the drug were noted. The results suggest that CZON is a useful antibiotic for obstetric and gynecologic infections.

[How to determine the prophylactic effect on postoperative infections].

With the cooperation of 13 medical institutions in the Tokyo area, the methods to determine the prophylactic effect on postoperative infections in gynecological surgery were evaluated. Two hundred and ninety-nine patients were enrolled for the study of postoperative infections, febrile morbidity and fever index following abdominal (275) and vaginal (24) hysterectomies. Prophylactic cefotiam (CTM) of 1 gram was intravenously administered twice a day postoperatively for 3 to 5 days. The rates of postoperative infections were 5.1% (14/275) in abdominal hysterectomy and 4.2% (1/24) in vaginal hysterectomy. The febrile morbidity (57.1% = 8/14) and fever index (52.3 +/- 41.1 degree hours) in the infection group were approximately about 4 times higher than those (12.3% = 32/261, and 15.6 +/- 13.7 degree hours, respectively) in the non-infection group. No significant differences were observed in age, body weight, height of patients, period of operation and blood loss between these 2 groups. These data suggested that febrile morbidity and fever index were able to indicate the prophylactic effect of antibiotics on patients undergoing abdominal and vaginal hysterectomies.