Fully automatic rapid DNA Ploidy Analyzer for intraoperative rapid diagnosis support.

Frozen section studies are a useful method to rapidly define tumor malignancy and identify the extent of surgical resection. However, diagnosis with a frozen section is qualitative and sometimes difficult. Therefore a quantitative method for grading tumors is desired. We have already reported a technique of intraoperative flow cytometry (iFC) that supports intraoperative histopathological examination of frozen sections. In this study, we report an advanced system named "Fully Automatic Rapid DNA Ploidy Analyzer" with a tissue pretreatment function and a freeze-dried reagent kit for cell staining. To evaluate our system, we analyzed samples from glioma patients who underwent open surgery for brain tumors. We observed obvious difference of the Malignancy Index (MI) between neoplastic and perilesional brain tissue (26.0 ±22.1% and 4.1 ±2.5%, respectively, P<0.001). Cut-off level for identification of the tumor in the biopsy specimen was 6.8% which provided 86% sensitivity and 81% specificity. We also obtained a good correlation between the MI and histological grade (WHO grading). Our new system also enabled finishing the process from sample preparation to the end of analysis in ten minutes or less. These results demonstrate that our fully automatic rapid DNA ploidy analyzer is feasible for rapid determination of glioma presence in a surgical biopsy sample.

Development of a de novo tumorous necrotic lesion in the liver after transcatheter arterial embolization combined with iodized oil infusion: report of a case.

We report herein the case of a 69-year-old woman in whom a hepatic tumorous necrotic lesion was discovered following transcatheter arterial embolization combined with iodized oil infusion (Lp-TAE) for a hepatoma. The lesion, which had not been evident prior to the Lp-TAE, was resected and analyzed pathologically. The portal area distribution in the necrotic lesion was the same as that in the surrounding hepatic tissue, suggesting that the lesion was derived from the nonneoplastic hepatic tissue. Moreover, extensive wall thickening and obstruction were observed in the intrahepatic portal vein and hepatic artery. These findings suggest that the lesion was a focus of hepatic infarction triggered by Lp-TAE.

Analysis of estrogen receptors in human breast cancer by assays using monoclonal antibodies and by the dextran-coated charcoal method.

Estrogen receptors (ER) were analyzed in 63 human breast cancers by immunocytochemical assay, and by enzyme-immunoassay, using monoclonal antibodies against human ER protein, and also by the dextran-coated charcoal method. The specific staining was observed only in the nucleus of cancer cells by the immunocytochemical assay, and the presence of nuclear staining by this assay was closely associated with the estimation of the cytosolic ER content by both the dextran-coated charcoal method and the enzyme-immunoassay. In the latter, the cytosolic ER content correspondingly increased with the nuclear ER content. Both the cytosolic and nuclear ER contents correlated well with those obtained by the dextran-coated charcoal method. Furthermore, the cytosolic ER contents obtained by the enzyme-immunoassay and the dextran-coated charcoal method correlated significantly with the age of the patient, but not with the nuclear ER contents. These above mentioned results suggest that ER is present mainly in the nucleus of breast cancer cells and that the cytosolic ER obtained by these two assays is the unoccupied one released from the nucleus.

Reduction of estrogen and prolactin receptors in 7,12-dimethylbenz(a)anthracene-induced rat mammary tumor by high doses of estrogen.

Daily injections of 100 micrograms 17 beta-estradiol, or 250 micrograms tamoxifen, for 10 days led to a regression of the 7,12-dimethylbenz(a)anthracene (DMBA)-induced rat mammary tumor. Estrogen receptor (ER), progesterone receptor (PgR), and prolactin receptor (PRL-R) in the regressed tumor were significantly reduced in the estrogen-treated rats. ER and PRL-R were low but PgR increased significantly in the tumor of the tamoxifen-treated rats. A single administration of 100 micrograms estradiol induced a transient decrease of ER and PRL-R, and an increase of PgR, in the DMBA-tumor. Similar decreases in ER and PRL-R and the increase of PgR were observed 8 hours after the 5th injection of 100 micrograms estradiol--a time when the tumor had already regressed. These results suggest that high dose-estrogen has a direct inhibitory effect on the concentration of both ER and PRL-R in the DMBA-tumor, and that this effect might be accumulative with repeated administrations. It is unlikely that the inhibition of the estrogenic effect caused by loss of ER is the sole mechanism of the regression of the DMBA-tumor, since the increased synthesis of PgR as a marker of estrogen action was observed even after the ER-reduction and tumor-regression.

Tamoxifen binding sites in human mammary cancers.

Tamoxifen binding sites (TBS) were measured using 3H-tamoxifen, the objective being to evaluate the relationships among TBS and hormone receptors and/or clinical and pathological characteristics in malignant tissues from 60 patients with mammary cancer. TBS were detected in most (96.7 per cent) cancers in the breast tissues, and the mean content and affinity were 569 fmol/mg X protein with Kd: 1.98 nM. There was no significant correlation between TBS and the estrogen receptor and/or progesterone receptor, with respect to positivity or content. However, there was a significant correlation between TBS and histological grading, thereby indicating the differentiation and the proliferative activity in this tissue. The content of TBS was significantly higher in the group with a high grade of malignancy. The TBS content significantly increased in parallel with the degree of malignancy, as related to tubule formation, nuclear pleomorphism and mitotic activity. On the other hand, there was no significant correlation between TBS and age, tumor size, lymph node status or clinical stage. These results suggest the possibility that TBS may be associated with differentiation and cell-proliferation in breast cancer tissues.

Isolation of a new procollagen V chain from chick embryo tendon.

Whole tendons of chick embryos synthesize procollagens V which consist of three pro-alpha chains: pro-alpha 1'(V), pro-alpha 1(V) and pro-alpha 2(V). This report shows that while the pro-alpha 1'(V) chain is similar to the pro-alpha 1(V) chain in many respects, such as similar but not identical peptide maps, it also distinctly differs from it in size and in other ways. The new chain is denoted as pro-alpha 1' to indicate the relationship. We have failed to see conversion of one chain into the other and they are regarded as variants, although we do not know whether they are different transcripts of one gene or products of two closely related genes. The pro-alpha(V) chains are assembled into the disulfide-linked homotrimer (pro-alpha 1'(V))3 and the heterotrimer [(pro-alpha 1'(V)S-S-pro-alpha 2(V))pro-alpha 1(V)] and a smaller amount of [(pro-alpha 1(V)2pro-alpha 2(V)]. The pro-alpha 1'(V) chains are processed similarly to the pro-alpha 1(V) by the initial removal of the presumed carboxyl propeptide yielding p-alpha 1'(V) and then by reduction in the size of the noncollagenous, presumed amino propeptide to yield alpha 1'(V). A size difference between the alpha 1'(V) and alpha 1(V) series of molecules is demonstrated by velocity sedimentation and by electrophoretic mobility of the reduced molecules. This difference is ascribed to a difference in the size of the propeptides because after pepsin digestion the products of both series of molecules are the same size and electrophorese like alpha 1(V)(pepsin). The carboxyl propeptides of pro-alpha 1(V) and pro-alpha 1'(V) are the same size, but the amino propeptide of pro-alpha 1'(V) is smaller than that of pro-alpha 1(V). The amino propeptide of pro-alpha 1'(V) and p-alpha 1'(V) also lacks asparagine-linked complex carbohydrate, which is linked to propeptides of the p-alpha 1(V) and p-alpha 2(V) chains. Differences between the alpha 1(V) and alpha 1'(V) series of molecules remain in material synthesized in the presence of tunicamycin. Primary cultures of tendon cells synthesize procollagen V consisting of the above three chains, but the procollagen V made by cultured tendon sheath synovial cells predominantly contains [(pro-alpha 1(V))2pro-alpha 2(V)].

Estrogen-dependent plasminogen activator in 7,12-dimethylbenz[a]anthracene-induced rat mammary tumors in vivo and in vitro.

Hormonal regulation of plasminogen activator in rat mammary tumor induced by 7,12-dimethylbenz[a]anthracene (DMBA) was studied both in vivo and in vitro. Plasminogen activator activity in DMBA-induced tumor (DMBA-tumor) was markedly decreased by ovariectomy, and recovered in a dose-dependent fashion upon estradiol administration, reaching a maximal level at 12 hr. This estrogen-stimulated production of the enzyme was prevented by actinomycin D, cycloheximide, and tamoxifen, indicating that in DMBA-tumor, estrogen might regulate de novo synthesis of plasminogen activator at a transcriptional level via an estrogen receptor system. Furthermore, DMBA-tumor cells in primary culture displayed similar estrogen-dependency toward the production of the enzyme without any cell proliferation. This indicates that the action of estrogen is mediated neither by cell division nor by prolactin, another hormone pastulated to be responsible for the development and growth of DMBA-tumor. Taken together, the present results have led to support the view that the primary function of estrogen is to induce plasminogen activator, which is probably essential to maintain the malignant state of DMBA-tumor.

[Biological and clinical analysis of breast cancer in relation to hormone receptors].

We examined whether or not the presence of hormone receptors (ERC, PRC, and ERN) were related to morphological and functional features in breast cancers. Well-differentiated tumors were more frequently receptor positive while poorly differentiated tumors were generally receptor negative. And labelling index and receptor contents were negatively correlated. There was a significant inverse correlation between the receptor contents and numbers of intracellular organellas in ultrastructural features of breast cancer cells. In LMI tests by 3-M KCl cancer extracts, migration of leucocyte from patients with poorly differentiated tumors were more frequently inhibited than those well differentiated tumors. Patients with receptor positive tumors were more endocrine sensitive and had better prognosis.