The growth and spontaneous dissemination of melanoma B16 and Lewis lung tumour in two sub-strains of C57BL/6J mice treated with Corynebacterium parvum and/or levamisole.

The effect of C. parvum and/or levamisole on tumour growth and spontaneous dissemination was tested in two tumour-host systems, melanoma B16 and LLT in C57BL/6J mice. C. parvum inhibited the growth of LLT and its dissemination to the lungs and the growth of B16 in females in one of the two sub-strains of mice used. Levamisole stimulated the growth of B16 in females in one sub-strain and in both sexes in the other sub-strain of mice. The growth of LLT was not influenced by levamisole but its dissemination to lymph nodes was facilitated. The growth-stimulating effect of levamisole was abrogated by C. parvum in the mice treated with both agents in combination. It is concluded that the effects of non-specific immunomodulators on tumour growth and spread can vary between sub-strains of the same inbred strain of mice bred at different laboratories.

ras p21 expression in relation to DNA ploidy, S-phase fraction and prognosis in colorectal adenocarcinoma.

ras p21 expression, as indicated by the monoclonal antibody ras 11, was estimated using immunohistochemistry on 69 primary colorectal adenocarcinomas. Also, DNA ploidy and S-phase fraction (SPF) were analysed with flow cytometry. Positive staining for ras 11 tended to be more common in DNA non-diploid tumours (P = 0.11), but was significantly correlated with high SPF (P = 0.038). Positive ras 11 staining, Dukes' stage, DNA ploidy and SPF were related to the recurrence-free interval of patients with Dukes' A-C tumours (P = 0.0014, P = 0.023, P = 0.035 and P = 0.040, respectively). ras 11 staining was a prognostic factor independent of both Dukes' stage and DNA ploidy (P = 0.011). The results indicate that pan ras p21 expression is associated with proliferative activity and has an independent prognostic value in colorectal adenocarcinoma.

Correlations of Ki-67 and PCNA to DNA ploidy, S-phase fraction and survival in uveal melanoma.

In 79 patients with uveal melanoma, the tumours were investigated by DNA flow cytometry and immunohistochemical staining of PCNA and Ki-67. S-phase as a continuous variable was significantly correlated with Ki-67 (P = 0.033), but not with PCNA. DNA ploidy was not correlated with either of the two antigens. Ki-67 was significantly correlated with histopathological type (P < 0.001) and tumour size (P < 0.001). Large tumours and epithelioid cell type were associated with a high frequency of Ki-67 positive cells. A high level of Ki-67 positivity (> or = 6.5%) was also associated with a shorter survival (P = 0.0037), and when adjusted for DNA ploidy, histopathological type and tumour size, Ki-67 in the multivariate analysis remained an important prognostic factor (P = 0.017).

Mucinous tumors of the vermiform appendix and ovary, and pseudomyxoma peritonei: histogenetic implications of cytokeratin 7 expression.

Cytokeratin 7 (CK-7) has been shown to be uncommonly expressed in colonic epithelial tumors, as opposed to ovarian epithelial tumors, which are always CK-7 positive. The authors investigated the expression of CK-7 in 17 appendiceal cystadenomas and carcinomas, 20 mucinous borderline tumors of the ovary, 10 cases of simultaneous mucinous tumors of the appendix and ovary, three so-called high-stage mucinous borderline tumors of the ovary, and three cases of pseudomyxoma peritonei (PP) of unknown origin. Nine appendiceal cystadenomas were CK-7 negative; two of these were associated with PP, and the peritoneal lesions were negative as well. Three cystadenomas were CK-7 positive. Three appendiceal carcinomas were CK-7 negative, and in one case the metastases were also negative. Two carcinomas were CK-7 positive. All 20 ovarian borderline tumors were CK-7 positive. Six cases of simultaneous mucinous tumors of the ovary and appendix were CK-7 negative, as were their peritoneal mucinous deposits. Four cases showed a positive reaction in both appendiceal and ovarian sites. Two of three so-called high-stage ovarian borderline tumors were CK-7 negative. All three cases of PP of unknown origin were CK-7 negative. In conclusion, appendiceal cystadenomas are often CK-7 negative, whereas ovarian mucinous borderline tumors are always CK-7 positive. The concordant staining pattern for CK-7 of simultaneous mucinous tumors involving the appendix and ovary (60% of which were CK-7 negative) supports an appendiceal origin for these tumors. Our results also support an appendiceal (or colonic) source for any CK-7-negative mucinous tumor involving the ovary or the peritoneum. Furthermore, our findings are in agreement with the assumption that mucinous borderline-like tumors in the ovary associated with PP are not ovarian in origin but are often, if not always, metastatic from an appendiceal (or other) mucinous tumor.

Interrelations of clinicopathologic variables and their prognostic value in colorectal adenocarcinoma.

We analyzed the interrelations of sex, age, tumor site, Dukes' stage, growth pattern and differentiation, and their prognostic value in 293 patients with primary colorectal adenocarcinoma. Simultaneously, growth pattern, differentiation, DNA and S-phase fraction (SPF) in paired primary tumors and lymph node metastases from 97 colorectal cancer patients were compared. The results revealed that poorly differentiated and mucinous tumors, as against well/moderately differentiated tumors, were frequently located in the proximal colon, and their frequencies were increased as Dukes' stage advanced (p=0.03). Tumor differentiation was usually identical in primaries and corresponding metastases (p=0.002), but this was not true of tumor growth pattern, DNA ploidy or SPF. In multivariate survival analyses, Dukes' stage provided strongly prognostic information (p<0.001) and mucinous tumors tended to predict worse survival (p=0.08).

Diagnosis and localization of prostate carcinoma by fine-needle aspiration cytology and correlation with histologic whole-organ sections after radical prostatectomy.

Twenty-nine patients with clinical stage T1-2, NO, MO prostate carcinoma were treated by retropubic radical prostatectomy. Diagnosis was made by fine-needle aspiration biopsy from six to eight separate sectors of the prostate. At the time of biopsy, diagrams of the palpated organ were drawn, depicting the location of the lesion and the site of each biopsy. Without the examiners' knowledge of cytologic data, extirpated prostate glands were examined with whole organ histologic sections, and carcinomas were scored according to the method of Gleason. The location and extent of all typical and malignant foci were mapped on a standard diagram. The results of preoperative cytologic examination were compared with postoperative histopathologic findings, showing a tendency toward underestimation of both the extent and degree of differentiation of the carcinomas during cytologic examination. In no case were these parameters overestimated during cytologic examination. The Gleason score correlated well with the presence of capsular and seminal vesicle invasion.

A comparison of three different methods for the determination of estrogen receptors in human breast cancer.

Forty-seven cases of breast cancer were analyzed for estrogen receptors (ERs) by three different techniques: (1) a biochemical assay using isoelectric focusing in polyacrylamide gel; (2) a monoclonal ER immunocytochemical assay (ERICA, Abbott Laboratories Diagnostics Division), and (3) a polyclonal antiestradiol immunohistochemical technique using formalin-fixed tissue. A high level of agreement (85%) was noted between the biochemical assay and ERICA. The ERICA-processed slides showed minimal background staining and were easy to interpret. Compared with the biochemical assay, ERICA achieved a positive predictive value of 86% and a negative predictive value of 82%. In contrast, results from the polyclonal antiestradiol histologic technique did not correlate with biochemical results and showed positive and negative predictive values of 61% and 25%, respectively. The authors conclude that the ERICA technique should prove to be a valuable adjunct to traditional biochemical ER assays.

DNA ploidy and S-phase fraction in primary melanomas and their regional metastases.

Flow cytometric analysis of DNA ploidy and S-phase fraction was performed on the primary melanomas and the first metastases from 55 melanoma patients with regional lymph node metastases or in transit metastases. The frequency of aneuploidy was significantly higher in metastases than in the primary tumour (p = 0.009), suggesting a higher growth potential in melanoma metastases than in the primary tumours. In 18 patients with reliable S-phase determinations from both primary tumour and metastasis there was no significant difference in mean S-phase fraction between primary melanomas and metastases. Skin metastases localized in dermis and subcutis had a significantly (p = 0.012) higher mean S-phase fraction than lymph node metastases.

Elective lymph node dissection in stage I cutaneous malignant melanoma of the head and neck. A report from the Swedish Melanoma Study Group.

The effect of elective lymph node dissection in patients with cutaneous malignant melanoma of the head and neck was investigated in a retrospective study. Of 517 patients in clinical stage I, 84 underwent elective dissection of the ipsilateral neck lymph nodes. In six of these patients, lymph node metastases were demonstrated at histopathological examination. There was a slight reduction in the incidence of recurrent disease in the regional lymph nodes in the group of patients who had undergone elective lymph node dissection, but this difference was not statistically significant. No significant differences were seen between the two groups regarding overall survival of disease-related survival.

Influence of aspirin and/or anabolic steroid (nandrolone decanoate) on experimental metastasis formation.

The influence on transpulmonary passage by an inhibitor of platelet aggregation, aspirin, and/or anabolic steroid, (Nandrolone decanoate), was investigated in syngeneic CBA mice. An increase of extrapulmonary metastases was used as a measure of increased redistribution of tumor cells from lungs to other organs. Aspirin alone did not reduce metastasis formation in lungs and did not significantly increase transpulmonary passage of tumor cells. The anabolic steroid increased the metastasis crop only in the lungs. However, treatment with the two compounds together increased the metastases in lungs and extrapulmonary organs. The results disclosed that in this system inhibition of platelet aggregation did not reduce metastasis formation but in combination with another drug, an anabolic steroid, it increased the number of experimental metastases.

Nuclear morphometry: a strong prognostic factor for survival after secondary surgery in advanced ovarian cancer.

Nuclear morphometry was performed on the diagnostic biopsy in 65 cases of non-mucinous ovarian carcinoma (FIGO stage IIB-IV) and its prognostic value regarding patient survival after the second-look operation was compared to that of morphology and clinical observations. In a univariate Cox survival analysis four morphometric factors were found to be significant predictors of survival (the standard deviations (SD) of the nuclear area, perimeter, largest perpendicular axis, and largest axis). Age, the size of residual tumor after the primary operation, and a combined variable describing the status at the second-look operation and also the result of tumor reduction were significant clinical variables. None of the morphologic variables proved to be significant. In the multivariate Cox analysis the SD of the largest perpendicular nuclear axis gave independent prognostic information together with either the size of residual tumor after the primary laparotomy (P = 0.00004) or the second-look variable (P < 0.00001). When the SD of the largest perpendicular nuclear axis and the second-look variables were included in the model the size of residual tumor after the primary operation added no further prognostic information. We conclude that nuclear morphometry is a simple, easily implemented and cheap quantitative method which gives objective and valuable prognostic information regarding survival in advanced ovarian cancer.

Experience with distant pathology demonstrations for clinicians in hospitals without local pathologists through the Swedish telepathology work station.

Conferences between pathologists and clinicians are routinely important at each hospital which has its own department of pathology. With the aid of telepathology, such conferences can be facilitated to provide results similar to those in hospitals with their own pathological service. Our Department serves the hospital in Motala, which is located 50 km west of Linköping. We now have telepathology conferences every second week with the surgeons and every second week with colleagues in internal medicine. The equipment we use is LabEye, which is produced by Innovativ Vision AB in Linköping. This equipment includes mouse pointers at both stations which are always visible in the images at both ends of the system. The quality of images received in Motala is very good, allowing discussion of details regarding the specimens, especially the cytological ones. Our experiences with these conferences have been very good.

The fat composition of a mouse diet modifies the effects of levamisol on growth and spread of a murine tumor.

The fat composition of the diet fed to C57BL/6J mice influenced the effect of Levamisol on the growth and spread of LLT. The timing of feeding of different diets as well as of the administration of Levamisol in relation to tumor transplantation was of importance. The tumor-suppressive effect of Levamisol given on day 9 was counteracted by a diet containing 2% soy oil instead of animal fat and given from day 1. Levamisol given on day 1 counteracted the facilitation of tumor growth and spread by the soy oil diet given from day 9. The soy oil diet was shown to suppress the immune response to SRBC. We conclude that standardization of the laboratory diets is necessary.

Tumour metastasis in mice with reduced immune reactivity. III. Studies with three weakly antigenic tumours in thymectomized and/or sub-lethally irradiated mice.

A previous observation that increased spontaneous metastasis formation from the highly antigenic MCA-induced sarcoma MCG101 occurred in immunosuppressed C57BL/6J mice prompted the present study to establish whether or not tumours capable of inducing only weak transplantation resistance would behave similarly. One MCA-induced sarcoma, MCG12, one epidermoid carcinoma, EpCa1 in CBA mice and a spontaneous mammary carcinoma, MaCa1 in C3H mice, were transplanted into adult thymectomized and/or sub-lethally irradiated mice at different intervals after irradiation. The effects of the treatments on the immunoreactivity were monitored in separate groups by determining the primary and secondary responses to SRBC and the survival of first set allogeneic skin grafts. The treatment resulted in a significantly increased incidence of mainly the lymph-node metastases with MCG12 and of only the lung metastases with EpCa1. No increase in metastases was noted with MaCa1. Thymectomy potentiated the increase in metastases from MCG12 in irradiated mice but unexpectedly so only for a limited time after irradiation. Local growth of MCG12 and MaCa1 at the transplantation site was more rapid in females than in males. The changes in tumour growth and spread induced by the treatments varied with tumour-host system, sex and time of transplantation after irradiation and did not conform well with any of the tested parameters of immunoreactivity. There was no close parallel between the effects of the treatments on growth and spread of the tumours.

Tumour metastasis in mice with reduced immune reactivity. IV. Restoration of relative resistance to metastasis formation in thymectomized sub-lethally irradiated mice by spleen cells.

Adult thymectomy and/or sub-lethal irradiation increased spontaneous metastasis formation in several syngeneic murine systems (MCA-induced sarcomas MCG101 in C57BL/6J mice, MCG12 in CBA mice and MCA-induced epidermoid carcinoma EpCa1 in CBA mice). In order to differentiate between immunological and non-immunological effects of these treatments on tumour spread the effects of presumed selective restoration of immune competence by transfer of syngeneic spleen cells from normal donors shortly after irradiation (1.5 months before tumour transplantation) was perfomed. This procedure brought the resistance of thymectomized irradiated mice to tumour spread back to the level seen in untreated mice, providing further evidence for the conclusion that the indrease of tumour spread in this experimental model to a major part was immune-mediated. In thymectomized irradiated mice adoptive transfer of concomitant immunity by spleen cells from tumour-bearing mice soon after tumour transplantation completely inhibited the spread of the highly antigenic MCG101 and reduced that of the weakly antigenic MCG12 to control level. In contrast, adoptive transfer after tumour resection had no significant effect of the spread from either to the tumours.