Measurement of circulating parathyroid hormone-related protein in rats with humoral hypercalcemia of malignancy using a two-site immunoradiometric assay.

The Rice H500 rat Leydig cell tumor is a well characterized model of humoral hypercalcemia of malignancy (HHM). Circulating concentrations of PTH-related protein (PTHRP) have not been reported in this or any other animal model of HHM. Taking advantage of the marked N-terminal amino acid homology between rodent and human PTHRPs, we have adapted a sensitive two-site immunoradiometric assay developed for measurement of human PTHRP for use in measuring rat PTHRP. Circulating calcium and PTHRP concentrations were serially measured after sc passage of the Leydig cell tumor in rats. Significant hypercalcemia and elevation of PTHRP occurred on day 9 after tumor inoculation. When grouped by tumor size, both plasma calcium and PTHRP levels were significantly elevated in animals with tumor burdens greater than 10 cc. The PTHRP concentration was strongly correlated with both serum calcium (r = 0.88) and tumor size (r = 0.80). Circulating rat PTHRP averaged 12.8 pM on day 9 and 27.5 pM on day 10 or 11. PTHRP was undetectable in the plasma of 19 control rats. In 3 rats, plasma calcium returned to normal, and PTHRP became undetectable within 24 h after tumor excision. Rat milk displayed a PTHRP concentration of 2000 pM, while acid-urea extract of the rat tumor contained 0.32 pmol/mg protein. Dilutions of rat plasma, milk, and tumor extract displayed response curves that were parallel to the human PTHRP-(1-74) standard in the assay. This two-site immunoradiometric assay is a sensitive and easily performed means of measuring rat PTHRP. It should be useful in studying this animal model of HHM and the function of PTHRP in normal tissues.

[Immunometric tests system for glycoprotein hormones and their free subunits]. / Immunometrische Testsysteme für Glykoproteinhormone und deren freie Untereinheiten.

Highly specific sensitive immunoenzymometric assays were established for human glycoprotein hormones, the common free alpha-subunit of glycoprotein hormones and the free beta-subunit of human chorionic gonadotropin, using a large panel of monoclonal antibodies previously produced in our laboratory. The significance of serum levels of human chorionic gonadotropin and its free subunits under physiological conditions, as well as their prognostic value in human chorionic gonadotropin-producing tumours was determined in vitro and in vivo. These investigations revealed that all three analytes were present in the sera of the majority of normal individuals; moreover, the percentage of positive sera for these tumour markers in patients suffering from seminomatous and non-seminomatous testicular cancers was significantly elevated when two, or even better, all three parameters were determined.

Changes in glycolipid expression in human testicular tumor.

Glycolipids were extracted from testicular tumor tissues of 13 patients, and their pattern of expression compared with that of normal testicular tissue. The most conspicuous and consistent change in the tumor extracts was marked accumulation of CTH (ceramide trihexoside). Structural analysis by enzyme cleavage showed that CTH which accumulated in the tumor tissue was Gb3 (Gal alpha 1-4Gal beta 1-4Glc beta 1-Cer). Immunohistochemistry using anti-Gb3 monoclonal antibody (MAb) (1A4) also indicated massive accumulation of Gb3 in the tumor tissue. Gb3 may be a new marker of testicular tumors, especially seminomas, for which useful markers are so far lacking.

Glial fibrillary acidic protein immunoreactivity of chondrocytes in immature and mature teratomas.

The immunoreactivity of chondrocytes for glial fibrillary acidic protein (GFAP), other intermediate filament proteins and S-100 protein was studied in formalin-fixed paraffin-embedded sections. A total of 95 cartilage specimens were examined from five immature teratomas, 12 mature teratomas, and a teratocarcinoma. GFAP-immunoreactive chondrocytes were abundant in immature cartilages, and as the cartilages maturated, these chondrocytes decreased and became distributed peripherally. Elastic cartilage had more GFAP-immunoreactive chondrocytes than non-elastic cartilage. GFAP-immunoreactive cartilage was often located close to central nervous tissue. Immunostaining for vimentin and S-100 protein revealed extensive distribution of immunoreactive chondrocytes in immature and mature cartilages, but in mature cartilage, chondrocytes at the center had less vimentin immunoreactivity. GFAP-immunoreactive chondrocytes also showed apparent immunostaining for vimentin. There was no difference in immunohistochemical staining for the alpha and beta subunits of S-100 protein. The immunoreactivities of teratoma cartilage specimens were quite similar to those of respiratory tract cartilage.

Nuclear deoxyribonucleic acid content (ploidy) of endodermal sinus (yolk sac) tumor.

Paraffin sections from 30 endodermal sinus (yolk sac) tumors were Feulgen-stained, and nuclear DNA content (ploidy) was assessed through DNA cytophotometry using the microTICAS image analysis system. The series consisted of 20 ovarian, 8 testicular, 1 sacrococcygeal, and 1 mediastinal tumors. Of the 30 endodermal sinus (yolk sac) tumors 29 were shown to be aneuploid, whereas the remaining tumor which exhibited the recently described primitive intestinal or enteric pattern was diploid and may thus be considered a neoplasm showing a somewhat higher degree of differentiation as compared to tumors showing the other histologic patterns of endodermal sinus (yolk sac) tumors.

Keratin 7 is a marker for a subset of trophoblastic cells in human germ cell tumors.

Human testicular germ cell tumors were studied immunohistochemically with the monoclonal antibody to the 54-kd keratin polypeptide (keratin 7) to determine whether this antibody could be used selectively to identify trophoblastic cells. The antibody reacted with the intermediate filaments in the cytoplasm of some cells in choriocarcinoma cell lines, and in trophoblastic cells in mixed germ cell tumors and a seminoma. It did not react with classic seminoma cells, embryonal carcinoma, yolk sac carcinoma, or somatic tissues of mixed germ cell tumors. On the basis of these data we conclude that monoclonal antibody to keratin 7 is a marker for a subset of trophoblastic cells in human germ cell tumors.

[Paratesticular angiosarcoma. Immunohistochemical confirmation. Review of the literature]. / Angiosarcoma paratesticular. Confirmación inmunohistoquímica. Revisión de la literatura.

We report a case of paratesticular angiosarcoma. To our knowledge, no well-documented case involving this site has been reported in the literature. The anatomopathological features of the present case are described, highlighting immunohistochemical analysis. The outstanding clinical features of this aggressive tumor type are discussed.

Isolation and characterization of a multipotent clone of human embryonal carcinoma cells.

Histopathological studies suggest that the stem cells of human teratomas may be classified into two major categories: nullipotent stem cells, and multipotent stem cells, capable both of self-renewal and differentiation into a wide range of somatic and extraembryonic cell types. We have isolated a multipotent stem cell clone from the human teratoma cell line GCT 27, and compared its properties to a nullipotent clone derived from the same strain. The multipotent clone GCT 27 X-1 gave rise to colonies of mixed cell morphology in vitro. Analysis of cell surface, cytostructural and extracellular matrix markers in GCT 27 X-1 cells showed that the stem cells of this line were very similar in phenotype to nullipotent cells. The two cell clones were predominantly hypotriploid, and contained several marker chromosomes in common. GCT 27 X-1 was feeder-cell-dependent for continuous growth in vitro; removal of the feeder layer resulted in differentiation of the stem cells into a variety of cell types, some with characteristics of extraembryonic endoderm, others showing neuronal properties. When transplanted into nude mice, GCT 27 X-1 cells gave rise to teratocarcinomas containing embryonal carcinoma stem cells, and many other cell types: yolk sac carcinoma cells; cells producing alphafetoprotein or human chorionic gonadotrophin; glandular, columnar, cuboidal, and squamous epithelium; primitive mesenchyme and cartilage; neuroectodermal cells. Nullipotent GCT 27 C-1 cells could form colonies in the absence of feeder layers, but multipotent GCT 27 X-1 cells could not. While a range of known growth factors and related substances failed to substitute for feeder layers in supporting the growth of GCT 27 X-1 stem cells, supernatants from yolk sac carcinoma cell line GCT 44 could partially replace the feeder cell requirement. Thus, the results revealed a basic difference in growth control between these multipotent and nullipotent human embryonal carcinoma cells, and suggested a possible paracrine regulatory pathway between multipotent stem cells and yolk sac carcinoma cells.

The pluripotential nature of the mesenchyme-like component of yolk sac tumor.

Germ cell neoplasms were reviewed for the investigation of a mesenchyme-like component of yolk sac tumor (YST) characterized by spindle cells with few mitoses in a myxoid, vascular background. Nineteen YSTs with this pattern were identified. The mesenchyme-like component of these YSTs appeared to derive from the epithelial elements of YST, since cytokeratin as well as vimentin positivity occurred in the spindle cells of the mesenchyme-like areas and foci of epithelial-spindle cell transition were present. In some cases the mesenchyme-like component showed differentiated mesenchymal elements (usually skeletal muscle). Similar features were identified in 13 chemotherapy-treated cases of YST that consisted only of this mesenchyme-like component. The mesenchyme-like component of YST appears to represent a chemoresistant, pluripotential cell population arising from metaplasia of YST epithelium; it may give rise to sarcomas occurring in some patients with treated germ cell tumors.

Adenomatoid tumor of the epididymis with special reference to immunohistochemical study of 3 cases.

Three cases of epididymal adenomatoid tumor are presented. The adenoid compositions of the tumors lined by epithelial cells showed a canalicular pattern with large vascular spaces, tubular pattern with glandlike regions or plexiform pattern with connective tissue strands. Immunohistochemistry demonstrated positive cytoplasmic staining for keratin, but negative for carcinoembryonic antigen and factor VIII-related antigen in each neoplastic tissue. These findings support the mesothelial origin of the epididymal adenomatoid tumors.

Adenocarcinoma of the rete testis with a spindle cell component. A possible metaplastic carcinoma.

A case of adenocarcinoma of the rete testis was encountered in a 36-year-old white man. The tumor fulfilled established criteria for determining origin in the rete and showed an unusual biphasic morphology with papillary adenocarcinoma mixed with a prominent component of cytologically malignant spindle cells. Immunohistochemical study demonstrated a positive reaction in the epithelium for cytokeratin and epithelial membrane antigen, and the cytoplasm of a few of the spindle cells also reacted with these antibodies. Electron microscopic study confirmed the biphasic pattern, showing epithelial gland formation and mesenchymal cells. The results indicate that this tumor is a metaplastic carcinoma of the rete testis. Recognition of this pattern of rete carcinoma may further enhance our knowledge of primary tumors at this unusual site.

Testicular plasmacytoma.

The case of a sixty-five-year-old man with multiple myeloma and a testicular plasmacytoma is described. This represents the thirty fourth reported case of testicular plasmacytoma and the first in which immunoperoxidase histochemistry has been used to demonstrate that the testicular plasma cells contain immunoglobulin of the same isotype as the patient's paraprotein. The clinical and morphologic features of previously reported testicular plasmacytoma are reviewed.

Disseminated primitive neuroectodermal tumor: diagnosis using immunocytochemistry, electron microscopy, and molecular probes.

A patient with a disseminated small cell tumor presented with hyperuricemia, gingival hypertrophy, lymphadenopathy, and bone marrow replacement with tumor cells. Initial histologic examination and clinical presentation were consistent with presumed marker silent lymphoma/leukemia. Despite initial treatment with and response to lymphoma/leukemia therapy the patient relapsed in the testis, bone marrow, pancreas, and skin whereupon subsequent and retrospective immunocytochemical, ultrastructural, cytogenetic, and molecular analysis led to the diagnosis of primitive neuroectodermal tumor (PNET). Despite extensive investigation and autopsy no primary site of tumor could be found demonstrating that PNET should be considered in the differential diagnosis of disseminated small cell tumors without an apparent primary.

[Clinical study of testicular tumor: tumor tissue content of hCG in seminoma].

Positive cases of serum human chorionic gonadotropin (hCG) have been increased in patients with pure seminoma due to the progress of methods of measuring serum hCG. But the positive ratio and the value of serum hCG in pure seminoma cases are lower than those of non-seminoma cases. The relationship between the pathological type of testicular tumor and the hCG levels in both serum and tumor tissue was investigated, and clinical usefulness of the serum hCG levels as a tumor marker of pure seminoma was discussed. The materials were 19 pure seminomas, 14 non-seminomas and 11 normal testes as controls. HCG and hCG-beta in tumor tissue extracts and in the serum of blood from the spermatic and peripheral veins were measured by radioimmunoassay (RIA). The tissue content of hCG and hCG-beta were 276.7 +/- 136.1 mIU/g.tissue and 16.5 +/- 3.20 ng/g.tissue, respectively, in pure seminoma; 224,376 +/- 91,619 mIU/g.tissue and 4,608 +/- 1,817 ng/g.tissue, respectively, in non-seminoma including choriocarcinoma component; 1,807 +/- 1,428 mIU/g.tissue, and 37.0 +/- 11.2 ng/g.tissue, respectively, in non-seminoma including seminoma component but not including choriocarcinoma component; 20.4 +/- 3.1 mIU/g.tissue and 1.27 +/- 0.38 ng/g.tissue, respectively, in normal testes, (each M +/- SE). The tissue content of hCG-beta in pure seminoma is significantly higher than that in normal testis (p less than 0.01) and significantly lower than that in non-seminoma including choriocarcinoma component.(ABSTRACT TRUNCATED AT 250 WORDS)

[Testicular germinal tumors. Current therapeutic principles]. / Tumeurs germinales testiculaires. Principes thérapeutiques actuels.

The cure rate for germinal tumors of the testes are now very high, greater than 80%, all stages and histological types taken as a whole. This calls for a revision of therapeutic indications, in light of these results and modern staging methods. In localized stages, or tumors associated with a good prognosis, therapeutic de-escalation is in progress, in order to limit the side effects. In invasive forms or forms associated with a poor prognosis, the use of more aggressive methods, such as high dose chemotherapy, should further improve prognosis.

Immunohistological determination of oestrogen receptor, progesterone receptor, and intermediate filaments in Leydig cell tumours, Leydig cell hyperplasia, and normal Leydig cells of the human testis.

Testicular Leydig cell tumours are able to produce oestrogens and can be induced by exogeneous oestrogen administration. Oestrogen and progesterone receptors, cytokeratin, vimentin, and proliferative activity were determined immunohistologically in human testes in six Leydig cell tumours, 14 cases of Leydig cell hyperplasia, and 13 cases with normal Leydig cells. While both steroid receptors were detected in about 70 per cent of the tumour cells in cryostat sections, no reaction was observed in normal Leydig cells. This supports the hypothesis of an enhanced receptor state in a Leydig cell subpopulation as a basic pathophysiological factor in the development of Leydig cell tumours. On cryostat sections, all tumours co-express cytokeratin and vimentin. Neither the receptors nor the intermediate filaments could be detected reliably in paraffin sections. The low proliferative activity of Leydig cell tumours corresponds to their benign clinical course.

[Pure testicular choriocarcinoma]. / Coriocarcinoma testicular puro.

We present a case of pure testicle choriocarcinoma in a 13-year-old male. Its first clinical manifestation consisted in a pattern of acute dyspnea caused by multiple pulmonary metastases. Alphafetoprotein investigation in serum and tissue proved negative. We carry out an immunohistochemical study of the case and discuss its histogenesis current classification and prognostic significance.

Identification and characterization of arginine vasopressin receptors in the clonal murine Leydig-derived TM3 cell line.

Specific arginine vasopressin (AVP) binding sites were identified and characterized using Leydig cell membranes prepared from a clonal murine Leydig-derived cell line, TM3. 3H-AVP binding data analyses demonstrated that the radioligand binds to a high affinity, low capacity, homogeneous class of sites with a dissociation constant of 0.5 nM. Characterization of these AVP binding sites included competition studies. Displacement of 3H-AVP binding with high affinity by unlabelled AVP, LVP and the V1 antagonist, d(CH2)5Tyr(Me)AVP, indicated that the Leydig cell AVP receptor is of the V1 type. Furthermore, AVP did not increase adenylate cyclase activity in TM3 membranes, a finding consistent with the V1 type of AVP receptor. No competition with 3H-AVP was found with the V2 agonist, dVDAVP, or the selective oxytocin agonist, [Thr4,Gly7]oxytocin. No specific binding for oxytocin was found in Leydig cell membranes. No specific binding for either 3H-AVP or 3H-oxytocin was observed in membranes prepared from the Sertoli cell line or peritubular cell line. These findings indicate that murine Leydig cells have specific AVP binding sites of the V1 type. These AVP sites are not coupled to the adenylate cyclase system.

Lectin histochemistry of human testicular germ cell tumors.

The lectin binding pattern (WGA, UEA-I, PNA, PSA, Con A, RCA and LCA) of 28 human testicular germ cell tumors (pure or combination tumors) was investigated. Lectin binding sites could be demonstrated in all germ cell tumor types. In classic and spermatocytic seminomas as well as seminomas with high mitotic index an equal distribution of lectin binding sites was observed. In embryonal carcinomas the lectin binding of UEA-I, PNA, WGA, LCA and RCA correlated to histological differentiation. A polarized staining of WGA, PNA, RCA and UEA-I, typical for embryonal carcinomas, yolk sac tumors and teratomas, was never seen in seminomatous tumors and may be of importance in differential diagnosis. By means of Con A decoration it was possible to distinguish cytotrophoblastic and syncytiotrophoblastic differentiation. Aspects of lectin histochemistry in tumor biology in general and in differential diagnosis of germ cell tumors in particular are discussed.