[Solitary tumor of the spermatic cord]. / Solitärer Tumor des Samenstrangs.

A 4.5-cm elastic, non-dolent mass in the left spermatic cord was diagnosed in a 57-year-old man and totally excised by a scrotal access. Histologically a cellular angiofibroma was diagnosed. These benign tumours are very rare. In women the most common sites are the vulvovaginal, in men the inguinoscrotal region.

Transmyocardial laser revascularization. Histological features in human nonresponder myocardium.

BACKGROUND: The creation of transmyocardial channels from the epicardium to the left ventricular cavity with the use of a CO2 laser is a modern approach in the treatment of patients with chronic ischemic cardiac disease. The histological features of human myocardium at different times after transmyocardial laser therapy have not been previously described. We had the opportunity to examine hearts from patients who died without clinical evidence of a persistent therapeutic effect at 3, 16, and 150 days after transmyocardial laser revascularization (TMR) respectively. METHODS AND RESULTS: We grossly localized the laser-created channels in unfixed and formalin-fixed tissue. Three ventricular levels were defined for cutting the hearts into four segments. Then, transmural blocks were excised and cut crosswise and lengthwise for histological investigation through the use of established staining methods. On day 3, laser-induced channels were filled with abundant granulocytes and thrombocytes, fibrinous network, and detritus and were surrounded by severe myocardial necrosis. Furthermore, the epicardial and endocardial portions were obstructed by fibrinous network and microclots. Granulocytes were mostly absent on day 16; in addition, the channels were filled with erythrocytes or fibrinous network. On day 150, we observed a string of cicatricial tissue admixed with a polymorphous blood-filled capillary network and small veins, which very rarely had a continuous wrinkled link to the left ventricular cavity. CONCLUSIONS: We found different stages of wound healing in human nonresponder myocardium after TMR, resulting in scarred tissue that displayed capillary network and dilated venules without evidence of patent and endothelialized laser-created channels. Experimental studies are necessary to analyze the morphological basis for TMR-mediated effects in human responder myocardium.

Phenotyping of human tumor-infiltrating lymphocytes before and after exposure to different in vitro stimulation conditions.

Tumor-infiltrating lymphocytes (TiL) and autologous peripheral blood lymphocytes (PBL), mainly from breast and kidney tumor patients were cultivated with high- and low-dose rIL-2 under addition of autologous tumor cells or nonmalignant epithelial cells. Tumor cells added to TIL-cultures induced an additional proliferative response. Before cultivation, CD8+ and CD25+ lymphocytes were more frequent in TIL when compared to PBL, while CD16+, CD19+ and CD56+ cells were rare. After culture, lymphocytes of both origins showed an increase of CD2+, CD25+, CD56+ and HLA-DR+ cells and a relative decrease of CD3+ cells. The CD4+/CD8+ ratios and a number of CD56+ cells were rIL-2 dose dependent.

Ki-67 immunostaining in human breast tumors and its relationship to prognosis.

Ki-67 labeling was quantified in 37 nonmalignant breast tissues and in 63 breast carcinomas by counting ten random high-power fields each in three section planes (RC) or by evaluation of the area with the highest labeling density (HDC). Both procedures proved to be highly correlated (rs = 0.94). Ki-67-positive fractions of the nonmalignant tissues (mean, 2.1% for RC and 4.1% for HDC) were significantly lower as compared with the carcinomas (mean, 14.5% for RC and 17.5% for HDC). In carcinomas the Ki-67 labeling was significantly associated with pT stage, axillary lymph node status, and tumor grading and inversely related to progesterone receptor status. Using the medians of both counting methods (12% for RC and 17% for HDC) as cutoff points, significantly different curves for overall and disease-free survival (median follow-up, 37 months) were obtained. However, Cox multivariate analysis failed to demonstrate an independent effect of Ki-67 labeling. In contrast, Ki-67 reactivity seems to be of independent prognostic value if a higher cutoff level was selected.

Study of the relationship between immunohistologically demonstrated lymphocytes infiltrating human breast carcinomas and patients' survival.

Eighty-five breast carcinomas were immunostained for CD3-, CD4-, CD8-, CD16-, CD22-, CD38- and CD57-positive lymphocyte subpopulations. The results were related to follow-up data (median follow-up 46 months) of 74 patients regarding overall survival and 73 patients in respect to disease-free survival. Whereas the number of axillary lymph node metastases (P less than 0.01) and the hormone receptor status (P less than 0.01) resulted in significantly different survival curves for overall survival, not one of the lymphocyte subset infiltrats correlated significantly which overall survival. For disease-free survival, pT stage (P less than 0.01) and nodal (P less than 0.01) and hormone receptor status (P less than 0.05) proved to be prognostically important. However, disease-free survival was not influenced by the infiltration of any lymphocyte subset.

In situ distribution of transforming growth factor alpha in normal human tissues and in malignant tumours of the ovary.

The distribution of transforming growth factor alpha (TGF-alpha) in human normal tissues from the uterus, Fallopian tube, ovary, small and large intestine, lung, spleen, kidney, and skin was studied by immunohistochemistry. TGF-alpha was found in epidermis, bronchial epithelium, intestinal mucosa, renal tubules, endo- as well as in exocervical and endometrial epithelium, and in the serous epithelium of the Fallopian tube. No TGF-alpha was detected in the stromal components of any of the tissues nor in any of the pre- and post-menopausal ovaries studied. Twenty-nine ovarian tumours including 23 ovarian carcinomas, one malignant mixed Mullerian tumour, two ovarian metastases of gastrointestinal carcinomas, one dysgerminoma, one sarcoma, and one fibroma were studied for TGF-alpha by the same immunohistochemical method. In 25 cases, specific cytoplasmic staining for TGF-alpha of epithelial tumour cells could be demonstrated. The pattern and intensity of the TGF-alpha immunostain varied among the TGF-alpha-positive tumours. No TGF-alpha was found by immunohistochemistry in the remaining four cases nor in the stromal tumour components of any of the lesions studied. Northern blot analysis for TGF-alpha mRNA was performed on 12 of the tumours. While the immunohistochemistry and blotting results correlated well in ten cases, discordant results were obtained in two lesions.

Lacking prognostic significance of beta 2-microglobulin, MHC class I and class II antigen expression in breast carcinomas.

To evaluate the impact of MHC antigen expression on the survival of patients with cancer, 77 human breast carcinomas were investigated for the expression of beta 2-microglobulin (beta 2m), HLA-A,B,C and HLA-DR. Thirty-one benign breast tumours were stained for comparison. The results for the carcinomas were related to the survival data of the cancer patients. The expression of beta 2m, HLA-A,B,C and HLA-DR was significantly lower in malignant tumours compared to the benign lesions. Whereas all benign tumours were positive for beta 2m and HLA-A,B,C and 28/31 positive for HLA-DR the following positivity rates were found in carcinomas: 74/77 for beta 2m, 57/77 for HLA-A,B,C and 10/77 for HLA-DR. The follow-up (median 45 months) of 66 cancer patients for overall survival and of 65 patients for disease-free survival revealed no influence of beta 2m, HLA-A,B,C or HLA-DR expression on the prognosis of this cancer. In conclusion, experimental data indicating the importance of MHC antigens in anti-tumour responses are not confirmed by the analysis of cancer patient survival data.

The expression of EGF receptors, EGF-like factors and c-myc in ovarian and cervical carcinomas and their potential clinical significance.

The expression of the epidermal growth factor receptor (EGF-R) and the c-myc oncogene was investigated in different specimens of ovarian and cervical carcinomas. The EGF-Rs were analyzed by EGF binding assay, immunohistochemistry and Northern blotting. For analysis of c-myc expression, we used Northern blotting and immunohistochemistry. Furthermore, tissue concentrations of EGF-like factors (EGF-F) were measured in the same tumor and non-malignant specimens. The biochemical determination of EGF-R demonstrated that EGF specific binding sites were detected in 36% of ovarian (n = 140) and 81% of cervical carcinomas (n = 42). High amounts of EGF-R (greater than 10 fmol/mg specific binding) were found in 8% of the ovarian and 41% of the cervical carcinomas. Increased expression of EGF-R specific mRNA was detectable in 7/21 ovarian and in 5/7 cervical carcinomas. A positive correlation between the amounts of EGF-R mRNA, the EGF-R binding data and the staining index of EGF-R immunohistochemistry was found. The EGF-R immunohistochemistry demonstrates that only the tumor cells produce increased amounts of EGF-R, while the stromal cells are EGF-R negative. Low amounts of EGF-R specific mRNA were also detected in biochemically EGF-R negative tumors. The c-myc specific mRNA signal was found in all cases investigated. It is shown that the c-myc expression was increased in 10/21 ovarian and 5/7 cervical carcinomas. There was no positive correlation between the amounts of EGF-R and c-myc mRNAs. The product of myc, as detected by immunohistochemistry, is found in tumor as well as in stromal cells. The levels of EGF-F were measured in extracts of 63 ovarian and 12 cervical carcinomas and in 21 non-malignant tissues. About 30% of the tumor extracts contained higher EGF-F levels (4-15 ng/mg) than those found in the non-malignant specimens. Tumors with high EGF-F levels expressed high amounts of c-myc RNA. The EGF-R status (n = 111) and the EGF-F levels (n = 63) were related to the prognosis of survival for patients with ovarian carcinomas. EGF-R positive (EGF-R(+)) ovarian carcinomas had a significantly higher response rate to chemotherapy. The survival time of the EGF-R(+) group is reduced compared to the EGF-R negative (EGF-R(-)) group if only patients in remission are used to construct survival curves. Furthermore, a poor prognosis for survival was noticed for ovarian carcinoma patients with high EGF-F levels.

Localization of human epidermal growth factor receptor in cervical intraepithelial neoplasias.

Immunohistochemical staining of epidermal growth factor receptor (EGF-R) with a monoclonal antibody was performed in 43 biopsies of cervical tissue. The distribution of the receptors in normal cervical tissue differs from that of cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma of the cervix. Whereas the staining reaction in normal squamous epithelium was confined to the basal and deep parabasal cell layer, in all cervical intraepithelial neoplasias, with or without human papilloma virus association, a homogeneous EGF-R staining reaction could be observed throughout the entire lesion. This means that the dysplasia cells of a CIN I-III, like the tumor cells of a squamous cell carcinoma, have a raised EGF-R content, which in the normal squamous epithelium is usually only found in the basal and deep parabasal cells that are capable of dividing. No EGF-R staining reaction could be detected in the higher, differentiated cell layers of the normal squamous epithelium of the cervix.

Immunohistological demonstration of lymphocyte surface antigens in postmortem lymphoid tissues.

The postmortem stability of cell antigens has hardly been studied. Using monoclonal antibodies (mabs) we examined the postmortem detectability of lymphocyte surface antigens in different lymphoid organs by comparing two sensitive, immunohistological staining procedures. To quantify the probable degree of autolysis of the tissues a score system was applied by taking into consideration the postmortem age as well as the core temperature of the corpses. The antigens examined generally proved to be very resistant to autolytic influences. Differences were found when comparing different mabs and with regard to the type of lymphoid tissue. The loss of immunohistological reactions was most extensive in the spleen whereas tonsils showed almost no qualitative alterations in staining patterns. Reactivity of mabs with postmortem tissues decreased in the following order: Dako CD22 and anti-Leu 4, anti-Leu 3a, anti-Leu 7, Dako T8. The mabs anti-Leu 7 and Dako-T8 frequently failed to demonstrate their respective antigens but no correlation between the loss of staining and the degree of autolytic decomposition (our score) could be detected. In general, postmortem tissues as well as tissues shock frozen after delay are suitable for qualitative immunohistology of those cells characterized by the mabs applied. The APAAP-method proved unequivocally to be the superior staining technique.