Lobular involvement: prognostic indicator for invasive ductal carcinoma of the breast.

One of the striking microscopic appearances of breast carcinoma is malignant transformation of the epithelium of an entire lobule, although the lobular architecture is still preserved. We termed this pathological feature 'lobular involvement'. An investigation was carried out to determine whether lobular involvement would be a useful prognostic indicator for invasive ductal carcinoma. One hundred and fifty cases of invasive ductal carcinoma were included in this study. The lobular involvement, as well as the conventional prognostic factors, was evaluated to find out the correlation with the overall and relapse-free survivals by univariate and multivariate analyses. The lobular involvement was identified in 31 of 150 patients (20.6%). Of the patients with lobular involvement, two had experienced recurrence (6.5%) and one had died (3.2%). The lobular involvement and lymph node metastases were significantly and independently correlated with the overall and relapse-free survivals. The presence of lobular involvement was statistically correlated with the favorable outcome. Lobular involvement can be assumed as a new prognostic indicator for invasive ductal carcinoma.

Enhancement of anti-herpetic activity of antisense phosphorothioate oligonucleotides 5' end modified with geraniol.

We have previously shown that antisense phosphorothioate oligonucleotide (SON) targeted against immediate early (IE) pre-mRNA5 of the herpes simplex virus type I (HSV-I) possessed potent anti-herpetic activities in vitro system. However, anti-herpetic activities of SON were not still efficient enough. Lipophilic compounds have been often conjugated with antisense oligonucleotide to enhance the biological activity. In this study, we selected geraniol as a lipophilic compound and newly synthesized SON bearing 5' terminal geraniol (geranyl-SON) toward IE pre-mRNA 5 of the HSV-1 to enhance the anti-herpetic activity. Geraniol is a olefinic terpene alcohol which is found in many essential oils. It possesses lipophilic characteristic. It is thought to be absorbed in tissue. Geraniol enhanced the anti-herpetic activity of SON with less cytotoxicity in a sequence specific manner. Terminal modification with geraniol did not affect binding affinity with complimentary DNA. Cytoplasm distribution of geranyl-SON was confirmed by confocal microscope. While some of the geranyl-SON was seen in the nucleus, unmodified SON had a punctate distribution in the cytoplasm with little in the nucleus. These results suggested that geranyl modification enhances anti-herpetic activity by changing the subcellular distribution of the oligonucleotides. Consequently geraniol-modifica-tion could provide new means for the efficient delivery of oligo-nucleotides.

Pancreatic arteriovenous malformation involving adjacent duodenum in a patient with gastrointestinal bleeding.

Gastrointestinal bleeding caused by arteriovenous malformation (AVM) of the pancreas is extremely rare. A 56-yr-old man with AVM of the head of the pancreas who was presented with recurrent episode of melena is described herein. The preoperative diagnosis was confirmed by angiographic study, and he was successfully treated with pancreatoduodenectomy. Histological examination of the resected specimen revealed pancreatic AVM involving the adjacent duodenal wall.

Correlation between angiographic features and pathologic changes in breast carcinoma.

The correlation between angiographic tumor blushing and underlying pathologic factors was studied. Twenty female patients with breast cancer were divided into three groups according to the degree of tumor blushing. Primary tumors were divided into three areas: marginal, central and intermediate areas. In each area, the number of blood vessels per 10(4) mu 2 was randomly counted in 10 fields and a mean value of vessels was compared among the three groups. It was found that the following pathologic findings were significantly related to strong angiographic staining: (i) well-circumscribed border; (ii) large tumor nest; (iii) severe inflammatory cell infiltration; (iv) mild stromal fibrosis; and (v) absence of lymph node metastasis. The mean value of blood vessels in the strongly positive group was significantly increased in the overall and the intermediate areas. The density of the vessels in the intermediate area was thus thought to be responsible for the tumor blushing.

Morphometric studies for objective diagnosis of intraductal carcinoma of the breast.

In order to obtain objective histologic criteria for distinguishing intraductal carcinomas of the breast from benign intraductal hyperplasia, a morphometric study was carried out on 15 cases of breast carcinoma (5 cases of comedo carcinoma, 5 cribriform carcinoma and 5 papillotubular carcinoma) and 10 benign control lesions (5 papilloma and 5 fibrocystic disease). Morphometric analysis was performed for each group with regard to 1) mean nuclear area, 2) mean nucleolar size, 3) mitotic index, and 4) nuclear count within predefined area (nuclear density) in histologic specimens. The results obtained were as follows: 1) Mean nuclear area for comedo carcinoma was distinctively large. This, together with its coefficiency variance, may be a good indicator for identifying this type of carcinoma. 2) Mean nucleolar size was significantly larger in each carcinoma group than in their benign counterparts. Nucleolar size was therefore considered to be the most reliable parameter for discriminating malignant cells from benign hyperplasia. 3) Mitotic index was relatively high in comedo and papillotubular carcinomas. 4) No difference was seen in nuclear density between the malignant and benign groups. In addition to the major criteria, combined application of these morphometric indicators might also be helpful for practical histologic assessment.

Lobular involvement in human breast carcinoma.

One hundred twenty-nine cases of breast carcinoma were examined in order to clarify the occurrence of lobular involvement with regard to stromal invasion. Lobular involvement was clearly recognized in 56 cases (43%) having discernible portions of intraductal carcinoma components. The process of lobular involvement was first recognizable as an extension of a few carcinoma cells from intralobular ducts into the acinar lumina. Progressive accumulation of the carcinoma cells resulted in marked swelling and distortion of the involved lobules. During the process, perilobular myoepithelial cells (actin-positive cells) were stretched and disappeared. Subsequently, breaks in the basement membranes were also observed, resulting in stromal invasion of carcinoma cells. The lobular involvement was classified into common and round varieties, possibly reflecting different growth activities of the carcinoma cells. The stromal invasion was initiated by peripheral budding or focal necrosis of the involved lobules. The latter pattern was often observed in comedo carcinoma. It was thus revealed that in breast carcinomas the terminal ductal-lobular units are quite often involved and can become sites of extraductal invasion.

A simple and sensitive radioimmunoassay for adenosine.

We developed a simple and sensitive radioimmunoassay (RIA) for adenosine. The RIA is based on the double antibody method with adenosine 2', 3'-0-disuccinyl-3-[125I]-iodotyrosine methyl ester (diSc-adenosine-[125I]-TME) as a tracer. Anti-adenosine antiserum for the RIA was raised in rabbits immunized with diSc-adenosine conjugated to human serum albumin (diSc-adenosine-HSA). All samples and standards were succinylated prior to assay. The present immunoassay allows detection of 6.25-400 pmol/ml of adenosine in sample. Values obtained by the RIA and by a HPLC analysis showed a high correlation with correlation coefficient of 0.997. In order to determine adenosine in plasmas, blood cells must be separated in the presence of 6 mM EDTA, 0.006% dipyridamole (Dip) and 23 microM 2'-deoxycoformycin (dCF) at 2 degrees C. Adenosine in plasma could be accurately determined by the proposed method even without any pretreatments by deproteinizing. The adenosine levels with or without EDTA-treated normal human plasmas determined were 26.2 +/- 7.26 and 100 +/- 3.62 pmol/ml (mean +/- SEM), respectively.

[Clinical studies using a highly sensitive radioimmunoassay for mid-region and carboxy terminus of parathyroid hormone in normal, hypo- and hypercalcemic states].

Parathyroid hormone radioimmunoassay (RIA), specific for mid-region of the PTH molecule, has been proven to be extremely useful for the differential diagnosis of abnormal calcium metabolism. Recently, we developed a highly sensitive RIA for PTH, consisting of PTH antiserum (CH9), 125I labelled Tyr42 hPTH (43-68) and synthetic hPTH (1-84) as standard. This RIA cross-reacted with mid-region and carboxyl terminals of PTH. The within-assay and between-assay coefficients of variation were less than 4.6% and less than 8.6%, respectively. The limit of detection was 50pg/ml. The levels of serum calcium, serum phosphate, serum creatinine, Tmpo4/GFR and creatinine clearance (Ccr) in normal healthy volunteers aged 20 to 50 years remained almost constant and showed 9.24 +/- 0.34mg/dl (mean +/- SD, n = 242), 3.34 +/- 0.38mg/dl (n = 242), 0.870 +/- 0.121mg/dl (n = 242), 3.20 +/- 0.54mg/dl GF (n = 189) and 103 +/- 17ml/min (n = 137), respectively. All healthy volunteers (n = 326) had measurements of PTH in the blood. From 20 to 50 years, normal PTH mean was 374 +/- 97pg/ml (+/- SD, n = 237) and ranged from 180-568pg/ml, and from 60 to 80 years it was 471 +/- 133pg/ml (n = 34) and ranged from 205-737pg/ml. Since we found that PTH was markedly elevated above normal when Ccr was below 40ml/min, and PTH was very significantly correlated with the reciprocal of Ccr (r = 0.8996, P less than 0.001) using a multivariate analysis, all of the patients whose Ccr was higher than 40ml/min were selected and examined in the following studies. Serum PTH values completely separated patients with surgically proven primary hyperparathyroidism (1 degree HPT) from malignant associated hypercalcemia (MAH), and patients with idiopathic hypoparathyroidism (IHP) from pseudohypoparathyroidism (PHP), both of which were diagnosed by Ellsworth-Howard test. PTH values in all of the patients with 1 degree HPT (n = 23) were above normal, but those with MAH (n = 6) were below the normal or lower normal range. PTH values in patients with PHP (n = 7) showed above normal, while those with IHP (n = 5) were below the normal range. PTH was normalized in post operative status in all patients after parathyroidectomy (n = 6). These results indicate that this PTH RIA is extremely useful for the differential diagnosis in diseases with calcium abnormalities.

Peliosis hepatis. An unusual case involving the spleen and lymph nodes.

A case of peliosis hepatis in a 52-year-old woman with unusual involvement of the spleen and lymph nodes is described. The initial sign was intraperitoneal hemorrhage from the splenic lesion, recessitating splenectomy. The hepatic lesion, which was first noticed during the operation, rapidly progressed and eventually resulted in hepatic rupture with fatal intraperitoneal hemorrhage. Serial sections treated by silver impregnation revealed degeneration and dissolution of the fine reticulin framework in the involved organs, suggesting the possible morphogenesis of the peliotic lesion. The patient had no history of any underlying disorders or of medication with steroids.

Hepatocellular carcinoma with rhabdomyoblastic differentiation.

An extremely unique case of a liver tumor occurring in a 70-year-old man is documented in this article. The primary tumor was well encapsulated by a thick, connective capsule and was histologically composed of two distinct elements, i.e., a common hepatocellular carcinoma (HCC) and a rhabdomyosarcoma. Metastasis of HCC was only seen in the left adrenal gland, whereas intrahepatic metastatic foci as well as tumor thrombi occluding the portal vein branches were composed exclusively of rhabdomyosarcoma. The possibility that the rhabdomyoblastic component might have come from the preexisting HCC by way of metaplastic proliferation is discussed.

The capacity for differentiation and production of human chorionic gonadotropin and alpha-fetoprotein in human embryonal carcinoma cell lines.

The capacity for differentiation of five human embryonal carcinoma (EC) cell lines was studied by examination of the morphology of xenograft tumors in nude mice. In addition to morphology, differentiation into extraembyronic elements was studied by examining the evidence of human chorionic gonadotropin (hCG) and alpha-fetoprotein (AFP) production. The EC cell lines can be divided into two groups: pluripotent (NEC 14 and Tera 2) and nullipotent (NEC 8, NEC 15, and ITO-II) cell lines. In nude mice, the pluripotent NEC 14 and Tera 2 cells produced EC mixed with various somatic elements such as glandular, neuronal, and cartilaginous tissue. However, typical extraembryonic elements, such as choriocarcinoma and yolk sac elements, were not usually observed. Production of hCG and/or AFP was identified in the tumors by both radioimmunoassay and the immunoperoxidase reaction. These results indicate that some human EC cell lines retain the capacity for differentiation into somatic and extraembryonic elements and that hCG and/or AFP appears to be produced by transitional types of cells that still retain the morphology of EC cells.

In vitro and in vivo studies of the production of placental proteins (HCG, SP1, PPs) in human choriocarcinoma cell lines.

The production of human chorionic gonadotropin (hCG), pregnancy-specific beta-1 glycoprotein (SP1), and the various placental tissue proteins (PP4, PP5, PP10, PP11, PP12, MP1) was examined in human choriocarcinoma cell lines using radioimmunoassay (RIA) and an immunoperoxidase (PAP) staining method. In 6 gestational and 2 nongestational choriocarcinoma cell lines, hCG was produced in vitro. A significant heterogeneity of hCG production was noted among the cell lines. Production of hCG in the xenograft tumors was consistent with the hCG production of the same cell lines in vitro. SP1 was demonstrated both in the culture supernatants and the xenograft tumors of 3 cell lines, and the production was found to be inversely correlated with the hCG production of the cell lines. Positive staining for PP4 and MP1, and weakly positive staining for PP5, PP10, and PP12 was found in the xenograft tumors by immunoperoxidase (PAP) stain, but the staining for PP11 was negative. These placental proteins showed positive staining in portions of choriocarcinoma cells. These results indicate that placental proteins are not equally produced in heterogeneous choriocarcinoma cells.

Effects of human interferons on human choriocarcinoma cells in vitro and in vivo.

Three kinds of human choriocarcinoma cell lines (BeWo, HCCM-5, and NUC-1) were used for examining the antiviral and antiproliferative activities of human interferons (IFNs) in vitro and in vivo. All of the cell lines showed only low sensitivity to the antiviral action of every IFN-alpha, IFN-beta, and IFN-gamma against vesicular stomatitis virus infection. However, 2-5A synthetase was normally induced by IFN-alpha in all of the cell lines. The [3H]thymidine incorporation of both BeWo and HCCM-5 cells was suppressed in dose-dependent manner at 48 hr after treatment with 1 to 1,000 units (U)/ml of IFN-alpha or IFN-beta and the growth of them was also slightly inhibited when treated continuously with 1,000 U/ml for 6 days in vitro. Another cell line NUC-1 was the least sensitive to these IFNs among the three cell lines. IFN-gamma did not show any antiproliferative effect on these cell lines. The intraperitoneal administration of 5000 or 10,000 U of IFN-beta suppressed the growth of xenografts developed in hamster cheek pouches and subcutis of nude mice when its administration was initiated on the first day of cell inoculation. These results indicate that although some heterogeneities exist among the cell lines choriocarcinoma cells are weakly sensitive to the antiproliferative activity of human IFNs.

[A new in vitro chemosensitivity test. Individualized chemotherapy against ovarian cancer and its clinical effect].

A new in vitro chemosensitivity test was developed from comparative studies on the cytotoxicity of anticancer drugs against human tumor tissues xenografted into nude mice and their cultivated cells in vitro. Half a gram of the material was sufficient to examine the sensitivity of the tissues to 10 kinds of potential anticancer drugs and the results were obtained within 24 hours. The test was applied to all of 20 patients with advanced ovarian cancer. The predictive accuracy was 58% in 12 evaluable patients. This response rate was higher than those of conventional combination chemotherapy with or without cisplatin and adriamycin. Individual ovarian cancers showed different sensitivities to the drugs. These results indicate that heterogeneity of sensitivity to anticancer drugs exists among individual ovarian cancers and that our new type of in vitro chemosensitivity test is useful for selecting the most effective drugs for each individual type of ovarian cancer.

Coexistence of adenocarcinoma and choriocarcinoma in the sigmoid colon.

A rare case of a colon carcinoma that developed in a 50-year-old woman is documented. Histologically, the removed tumor was composed of two distinctive elements, i.e., well-differentiated papillary adenocarcinoma and choriocarcinoma. It was strongly suggested that the latter malignancy derived from the pre-existing colonic carcinoma through metaplastic proliferation. A very high human chorionic gonadotropin level was noted in the patient's serum obtained at the preoperative stage. Autopsy materials revealed that metastatic foci in the liver, lungs, and lymph nodes were exclusively composed of choriocarcinoma.

Establishment and properties of a human choriocarcinoma cell line of ovarian origin.

A human nongestational choriocarcinoma cell line of ovarian origin (IMa) was established in vitro. This cell line had been subcultured serially more than 22 times over 18 months. Small polygonal cells with a prominent nucleus were dominant and a sparsity of cytoplasmic organelles was an ultrastructural characteristic of the IMa cells. The production and secretion of human chorionic gonadotropin and its subunits were identified by radioimmunoassay. The IMa cells were transplantable in the hamster cheek pouch and the histological diagnosis was choriocarcinoma. A newly established ovarian choriocarcinoma cell line can be considered useful for clarifying the biological differences between nongestational and gestational choriocarcinoma cells.

A newly established human gestational choriocarcinoma cell line and its characterization.

The cell biological characteristics of a newly established human gestational choriocarcinoma cell line, HM, were examined and compared with four other gestational and two nongestational choriocarcinoma cell lines. Striking heterogeneity in the production and secretion of hCG and its subunits was observed among the cell lines. The HM cell line consisted mainly of cytotrophoblast-like cells, and the hCG and its subunit values in both the cells and culture fluid were extremely low. The syncytiotrophoblast-like multinucleated giant cells which were occasionally seen in other cell lines were scarce in the HM cell line in vitro. On the other hand, deviation of the LDH isozyme pattern to the heart-type (LDH1) was characteristic of the HM cells in contrast to the other cell lines in which LDH3 was dominant. The HM cell line represents useful material for investigating the cell biological heterogeneity of human choriocarcinomas.