Rapid, complete and reversible transformation by v-sis precedes irreversible transformation.

v-sis is the oncogene of simian sarcoma virus, but whether tumor growth is maintained by v-sis expression alone or requires additional changes is unknown. To distinguish these possibilities we studied a model of reversible transformation including tumorigenicity using NIH3T3 cells bearing a metallothionein promoter-v-sis construction. Cells subcultured from 10 out of 18 tumors from athymic mice, all less than 0.1 g and less than or equal to 21 days in age, reverted to a normal phenotype but exhibited transformation upon addition of zinc as judged by morphology, growth rate, saturation density and anchorage independence of growth. Thus, activation of v-sis alone is sufficient for initiation and early autocrine-based growth of tumors. However, the cells from the remaining and predominantly larger, 0.5 +/- 0.7 g, tumors did not revert and exhibited zinc-independent transformation as judged by the same criteria. Southern analysis and examination of the regulation of v-sis product expression in cells derived from these tumors showed no change in zinc-dependent and reversible regulation of v-sis sequences. These results suggest that subsequent tumor growth strongly favors acquisition of additional irreversible change(s) in the tumor cell genome at high frequency (44%). Thus an early event of a multistep process stimulated by v-sis-dependent transformation best accounts for the sum of results.

The proto-oncogene c-fos is over-expressed in the majority of human osteosarcomas.

The expression of fos protein was examined in 30 cases of human osteosarcoma as formalin-fixed and paraffin-embedded tissue sections using two monoclonal antibodies and, for five cases, as frozen sections using 3 polyclonal antibodies. Nuclear antibody labeling intensities were determined either by visual scoring (3 pathologists) or by microdensitometry and included over 700 tumor, 350 normal and 150 benign tissue observations. Visual scores were shown to be linear with optical density with a correlation coefficient of 0.97. Analysis of the osteosarcoma cases at one antibody concentration revealed two groups: a minority (39%) with a low average visual score of 2 +/- 0.2 which was very similar to benign and normal tissues, and a majority (61%) with an average score of 3.1 +/- 0.3. The difference is highly significant (t-test), P less than or equal to 0.01. The results were supported by an analysis of partial immunotitration curves using a curve-fitting procedure which yielded estimates of FOSo (maximum relative fos protein concentration) which were ca. 150% increased for the majority tumor group compared to the minority group or benign or normal tissues. In previous studies of v-sis transformed cells, which exhibit ca. 300% over-expression of c-fos protein as observed here, antisense techniques were used to show that over-expression leads to increased growth and loss of contact inhibition. Thus the combined results suggest that steady state c-fos protein is significantly elevated in, and may contribute to, the aggressive growth properties of a majority of human osteosarcomas.

C1qR(P), a myeloid cell receptor in blood, is predominantly expressed on endothelial cells in human tissue.

C1qR(P) is a type I cell surface glycoprotein that has been shown to enhance ingestion of suboptimally opsonized targets by phagocytes in vitro. In this study, we developed and characterized polyclonal antibodies to study the tissue distribution of this receptor targeted to either the N- or C-terminal portion of the molecule. C1qR(P) was detected in vascular endothelial cells and in a subset of pyramidal neurons in the brain, as well as neutrophils, but it was absent in most tissue macrophages. Analysis of in vitro differentiation of blood monocytes to dendritic cells demonstrated a down-regulation of the receptor as monocytes differentiate to dendritic cells, providing a possible explanation for the lack of reactivity of these cells in tissue. The predominant presence of C1qR(P) in endothelial cells, while compatible with a phagocytic role in host defense and/or clearance of cellular material, suggests other possible novel roles for this receptor.

Massive pneumatosis without necrosis: A case report of Clostridium perfringens sepsis in an extremely low birth weight infant.

Pneumatosis intestinalis and free intraperitoneal air on abdominal radiographs are considered pathognomonic signs of necrotizing enterocolitis (NEC). We report a unique case of late-onset fulminant sepsis due to Clostridium perfringens presenting with shock, extensive pneumatosis intestinalis and free intraperitoneal air in an extremely low birth weight infant without histopathological evidence of bowel necrosis or NEC.

Acute ascending necrotizing myelopathy caused by herpes simplex virus type 2.

A 57-year-old diabetic man died of a rapidly ascending necrotizing myelitis. Autopsy results proved that the etiologic agent was herpes simplex virus type 2 (HSV2). The clinical findings, autopsy immunohistopathology, and electron-microscopy suggest that either primary HSV2 infection or reactivation of HSV2 infection within dorsal root ganglia was followed by spread to the spinal cord. Viral infection of the CNS occurred by direct extension and led to death by involvement of the brainstem. Although there is only one previous report of HSV2 myelitis in the literature, our findings suggest that HSV2 might be a more common etiologic agent of necrotizing myelitis. Because CSF cultures are usually negative, viral inclusions are not usually seen, and morphologically identifiable virions are exquisitely rare, previous cases were probably descriptively diagnosed as acute ascending necrotizing myelitis without etiologic identification.

Trisomy 11 and other nonrandom trisomies in congenital fibrosarcoma.

Chromosome studies in a 7-week-old female infant with an intraabdominal malignant fibrosarcoma showed a hyperdiploid karyotype of 50,XX, +der(6)del(6)(p23)add(6)(q11), +8, +10, +11,add(12)(p13). Trisomy 11 appears to be a nonrandom primary cytogenetic abnormality in the congenital or infantile form of this mesenchymal tumor and is also a nonrandom gain in congenital mesoblastic nephroma. A possible developmental link between these two mesenchymal tumors, mediated by a gene or genes on chromosome 11 is postulated.

Unbalanced translocation resulting in the loss of the chromosome 17 short arm in an osteoblastoma.

We report results of the first cytogenetic study of an osteoblastoma. Cells from the tumor were characterized by a three-way unbalanced translocation involving chromosomes 15, 17, and 20. As a consequence of the translocation, most of one chromosome 17 short arm appears to have been lost. This loss suggests the possibility that loss of 17p DNA sequences may be involved in early changes leading to neoplastic proliferation of osteoblasts.

Presence or absence of trisomy 11 is correlated with histologic subtype in congenital mesoblastic nephroma.

Fluorescence in situ hybridization utilizing a probe for the alpha satellite repeat sequence on chromosome 11 was used to detect variations in the number of chromosomes 11 in 24 formalin-fixed, paraffin-embedded congenital mesoblastic nephromas. Evidence of trisomy 11 was found in nearly half of the tumors. More importantly, the presence of trisomy 11 was associated with the cellular histologic variant of this tumor.

Mammary epithelium-induced motility of MCF-7 cells.

BACKGROUND: Cancer cell invasion may be mediated by motility factors elaborated by the surrounding normal host tissue. This study was performed to determine whether normal breast cells induce motility in breast cancer. MATERIALS AND METHODS: MCF-7 breast carcinoma cells were co-cultured with either normal cultured human mammary epithelial cells (HMEC) or immortalized 184 A1 mammary epithelial cells and observed for evidence of motility. The ability of conditioned medium from mammary epithelial cells to induce motility in MCF-7 was measured with scattering assays, Boyden chamber assays and time-lapse video microscopy. RESULTS: A soluble factor in the conditioned medium of both 184 A1 and HMEC induced motility in MCF-7. The motility factor was trypsin-sensitive, but activity remained after 5 minutes of boiling or 2 hours at pH 2. CONCLUSION: Mammary epithelium secretes a protein capable of inducing motility in breast cancer cells, raising the possibility that this effect contributes to the invasive properties of human mammary carcinoma.

Granular cells in a cellular neurilemmoma.

In this article, we describe a case of a cellular neurilemmoma with focal granular cell elements. The Schwann cells showed S100 and Leu-7 immunoreactivity. The granular cells were periodic acid-Schiff-positive, diastase resistant with strong S100 immunoreactivity. Electron microscopy showed electron dense, heterogeneous granules in the granular cells and interdigitating processes and elongated nuclei of the Schwann cells. Although many granular cell tumors are believed to be derived from Schwann cells, neoplasms with both elements are rarely presented.

Aneurysm of the mitral valve in a patient with hypertrophic cardiomyopathy.

We report a case of mitral valve aneurysm and severe mitral regurgitation complicating infective endocarditis in a patient with hypertrophic cardiomyopathy. 2-dimensional echocardiography revealed a saccular structure in the anterior mitral leaflet that bulged into the left atrium throughout the cardiac cycle. Pathology of the excised valve showed inflammation, early repair and perforation of the aneurysm. Concurrent mitral insufficiency or trauma resulting from septal-anterior mitral leaflet contact may predispose to valvular infection. The repair process of this damaged focus and the loss of the elastic properties of the valve may contribute to aneurysm formation. The aneurysm in this case formed in less than 5 weeks.

Reversible transformation by v-sis: a model cell line for analysis of transformation by antisense methods.

Cellular homologs of v-sis are implicated in numerous human tumor types (Eva et al., 1982; Betsholtz et al., 1984; Johnson et al., 1985; Bronzert et al., 1987; Igarashi et al., 1987; Nister et al., 1988a,b; Versnel et al., 1988; Matsui et al., 1989), but whether tumor growth is maintained by sis expression alone or requires additional changes is unknown. To distinguish these possibilities, we studied reversible transformation of NIH-3T3 cells bearing an inducible v-sis construction. Cells subcultured from 10 of 18 tumors, all less than 0.1 gram and less than or equal to 21 days in age, reverted to a normal phenotype by four criteria, but again exhibited transformation upon induction. Thus, activation of the v-sis autocrine loop alone is sufficient for initiation of tumors. Cells from the remaining and larger, 0.5 +/- 0.7 gram, tumors did not revert by any criteria. This suggests that subsequent tumor growth is maintained by acquisition of irreversible change(s) that occurs at high frequency in vivo.

Transplacental hemorrhage associated with placental neoplasms.

We report a case of choriocarcinoma in situ arising from a term placenta in an otherwise normal pregnancy that resulted in fetal hydrops and intrauterine fetal death from chronic fetal-maternal hemorrhage (FMH). The clinical and pathologic features are described and compared with the few similar cases reported and with an additional placental choriocarcinoma found in our files. We also describe the clinical and pathologic observations of two chorangiomas that caused massive FMH and led to fetal death.

Lymphocyte and monocyte-induced motility of MCF-7 cells by tumor necrosis factor-alpha.

A potentially important tumor-host interaction is increased tumor-cell invasiveness in response to motility factors derived from stromal and lymphoid cells. Conditioned medium of IL-2-stimulated lymphocytes and fractions enriched in either T cells, natural killer (NK) cells, or monocytes induced motility in MCF-7 breast carcinoma cells. ELISA and antibody neutralization studies demonstrated that this effect was due to tumor necrosis factor-alpha (TNF-alpha) secretion by the lymphoid cells or the enriched fractions. Unstimulated leukocytes in direct contact with MCF-7 cells also induced motility that was inhibited by anti-TNF-alpha antiserum. Time-lapse video microscopy of cells exposed to 10 ng/ml TNF-alpha showed that motility was independent of its toxic effects. Immunoperoxidase showed that MCF-7 cells expressed both the 55-kDa and the 75-kDa TNF-alpha receptors (TNFR). Antiserum against the 55-kDa TNFR, like TNF-alpha, induced motility in MCF-7 cells. This was most likely due to cross-linking of the 55-kDa TNFR monomers, since the monomeric F(ab) did not produce this effect. Our results raise the possibility that TNF-alpha-induced motility is one mechanism by which tumor cells overcome the potential anti-tumor immune function of lymphocytes and macrophages in peri-tumoral infiltrates.

Congenital mesoblastic nephroma: cytogenetic comparison to leiomyoma.

We report the karyotype analysis of a congenital mesoblastic nephroma (CMN), a usually benign renal tumor occurring most commonly during early infancy. The tumor was composed of interlacing bundles of spindle-shaped cells and it displayed both the classic and cellular histologic patterns. Immunoperoxidase studies showed reactivity for vimentin and actin. The tumor cell karyotype included additional chromosomes 11 and an altered chromosome 12. Trisomy or tetrasomy of chromosome 11 is present in five of six reported cases and may represent a frequent alteration in the karyotype of the CMN. Furthermore, breakpoints in the q13-15 region of chromosome 12 are commonly seen in leiomyomas, which are histologically similar to the CMN. Thus, the karyotype described here illustrates another similarity between the two tumor types.

A report of two cases of recurrent Paget's disease of the vulva in a split-thickness graft and its possible pathogenesis-labeled "retrodissemination".

Two cases of recurrent noninvasive Paget's disease of the vulva in a split-thickness graft without an underlying adenocarcinoma are presented. This is the third report of recurrence of extramammary Paget's disease in a split-thickness graft, and the second of such an occurrence without an underlying dermal adnexa adenocarcinoma. A hypothesis for the possible pathogenetic mechanism of this unusual biological behavior is suggested.

Characterization of N-ethyl-N-nitrosourea-induced malignant and benign breast tumors in rats by using three MR contrast agents.

A carcinogen (N-ethyl-N-nitrosourea)-induced animal tumor model was established to grow malignant and benign breast tumors. In each tumor the pharmacokinetic characteristics were measured by using three contrast agents, gadolinium-diethylene-triamine-pentaacetic acid (Gd-DTPA; <1 kD), Gadomer-17 (35 kD), and albumin-Gd-DTPA (70-90 kD). Infiltrating ductal carcinomas (IDC) with low, medium, and high Scarf-Bloom-Richardson grades and fibroadenomas (FA) were analyzed. We found that Gd-DTPA could differentiate between FA and malignant tumors, but not between malignant tumors of low and high grades. In contrast, the intermediate size agent Gadomer-17 could differentiate between high-grade and low-grade IDC, but not between low-grade IDC and FA due to their similar enhancement patterns (despite their different origins). The largest agent, albumin-Gd-DTPA, was capable of differentiating both, but the low contrast-to-noise ratio was its major technical concern. The results in this breast tumor model suggest that macromolecular agents provide useful information for differential diagnosis among IDCs of various grades, but they do not provide superior information than Gd-DTPA for differential diagnosis between IDC and FA.

Radiation-induced CA 125 production by mesothelial cells.

CA 125 levels are often falsely elevated in disease-free endometrial cancer patients who have undergone abdominal radiation therapy. Because peritoneal irritation or mediators of inflammation can induce CA 125 production in mesothelium, the possibility that irradiated cultured mesothelial cells secrete CA 125 was investigated. Seven mesothelial cell isolates, an ovarian cell line which does not secrete CA 125, normal mammary epithelium, and normal fibroblasts were exposed to 500 cGy of 6-MV photon irradiation. Irradiated mesothelial cells showed little or no growth, while untreated cells increased in number. Twenty-four-hour CA 125 production was measured in the tissue culture medium on Day 4, and daily for one mesothelial cell isolate. Radiation stimulated CA 125 secretion in mesothelial cells up to 32 times over nonirradiated controls. The time course study showed that CA 125 levels increased rapidly in irradiated cells by Day 3 and remained elevated for the next 3 days. Increased immunoreactivity for p53 in irradiated mesothelial cells confirmed that a protein known to be radiation-inducible could be produced by the same conditions. Normal fibroblasts, mammary epithelium, and the ovarian cell line did not produce CA 125 in either the presence or absence of radiation. Thus, irradiated mesothelial cells are a potential source of serum CA 125 in patients who have received abdominal irradiation.

UCI-VULV-1, a vulvar squamous carcinoma cell line.

Squamous carcinoma of the vulva (SCV) is an uncommon neoplasm of uncertain etiology. There is evidence that there are two subgroups of SCV, one associated with human papilloma virus (HPV) and a second HPV-negative group. The UCI-VULV-1 cell line, obtained from a lymph node metastasis of an SCV, grows with a population doubling time of approximately 60 hr. The saturation density is 10(5) cells/cm2. The cell line does not exhibit anchorage independence and is weakly tumorigenic. The cells range in appearance from an abundant spindle cell to a less common larger, flat cell. All of the cells are immunoreactive for high-molecular-weight keratin, but only the flat cells, which form squamous pearls in vivo, are immunoreactive for low-molecular-weight keratin. The cell line expresses epidermal growth factor (EGF), transforming growth factor-alpha, the EGF receptor, and p53 protein. Polymerase chain reaction revealed no HPV DNA within the cells. Early passage cells exhibited karyotypic heterogeneity with few similarities to previous described SCV karyotypes. The cells display sensitivity to cis-platinum in concentrations toxic to many ovarian and cervical carcinoma lines. UCI-VULV-1 may be helpful for studying the properties of the HPV-negative form of SCV.