The value of cell proliferation and angiogenesis in the prognostic assessment of ovarian granulosa cell tumors.

OBJECTIVES: Most cases of granulosa cell tumors (GCT) of the ovary are characterized by a relatively good outcome. However, some tumors behave aggressively and some tend to recur many years after the initial diagnosis. Tumor growth depends on cell proliferation and angiogenesis. Thus, proliferative indices and microvessel density were studied to determine possible valuable methods to assess the GCT patient's outcome. METHODS AND STUDY DESIGN: Paraffin-embedded tissue blocks were available for 60 patients with primary GCT and were investigated by immunostaining with monoclonal antibodies against PCNA, Ki-67 and factor VIII-related antigen. The follow-up was available for 51 patients and ranged from 25 to 206 months. A clinical follow-up distribution of patients was made: 8 patients with recurrence (group I); 6 patients who lived with no evidence of recurrence for 100 months or more (group II), and 37 patients alive with no evidence of recurrence in the follow-up period of less than 100 months (group III). RESULTS: There was a statistical correlation between PCNA and Ki-67 proliferative indices. A significant increase (P <0.05) of mean PCNA and Ki-67 proliferative indices and mean tumor size was seen in patients of Group I compared to those of Group II. The mean PCNA proliferative index positively correlated with the mean Ki-67 proliferative index for Groups I and II. Mean microvessel density showed a positive correlation with mean PCNA and Ki-67 proliferative indices and with mean tumor size for Group I, whereas it was negatively correlated with PCNA proliferative index and tumor size for Group II. A positive correlation was found between mean mitotic count and both proliferative indices only for Group II. The following features were indicative of a relatively poor prognosis: GCT measuring >9 cm in diameter, PCNA >4.0%, Ki-67 >1.2%, and diffuse, insular and sarcomatoid histologic patterns. CONCLUSIONS: The findings support the importance of proliferative factors, tumor size and histologic patterns as possible prognostic indicators for estimating the biologic behavior of patients with GCT. Unfortunately, angiogenesis did not seem to be a useful determinant parameter of a possible aggressive behavior. However, a longer follow-up period with larger series may be required to assess the value of the parameters in prediction of patient survival.

The life-shortening effect of reduced physical activity is abolished by a fat rich diet.

In female mice on a control diet (3.6% fat) reduced physical activity leads to a reduction of the average life span. So the average age at death of an inactive group is 500 +/- 166 compared to 565 +/- 175 days in an active control group. If the animals are kept on a fat rich diet (12.4% fat) this effect of physical activity restriction is no longer observable and the average age at death is 570 +/- 142 days, within the range of the control animals. The increased fat intake seems to reduce the stress or to increase the resistance to stress in the activity restricted animals. So stress is a crucial determinant of life span.

A glucose-rich diet shortens longevity of mice.

High plasma levels of glucose and insulin over long-time periods play an important role in the genesis of diabetic complications. There is evidence that the long term consumption of glucose-rich diet by rats is detrimental to insulin sensitivity. We investigated the effect of a glucose-rich diet on longevity of 70 female mice which were compared to 70 mice on a control diet. The average age of death of the control group was 568 +/- 139 days compared to 511 +/- 170 for the glucose group and the seven oldest mice of the control group died at age 890 +/- 52 days, while the seven oldest mice of the glucose group died at 833 +/- 49 days. These differences are statistically significant (P < or = 0.05). Our work shows that a life-long intake of a diet with 20% of total energy derived from glucose leads to a significant reduction of the average and maximal life-span in female mice and thus, supports previous observations of detrimental effects of high glucose intake over long periods.

Systemic application of cysteine-2:1-crotonal adduct for pharmacokinetic studies of tumor models in rats and mice.

A selective cytostatic effect is demonstrated for systemic application of the Michael adduct of crotonal (CAS 4170-30-3) with cysteine (CAS 52-90-4). As in earlier studies, our initial model was the Ehrlich ascites tumor (EAT) in mouse. Tablets containing 150 mg of the substance were implanted subcutaneously. Substance concentrations were found to be higher in the ascites fluid than in the blood. A better model, the Walker-256 carcinosarcoma (Wa256), was used subsequently. Due to its rapid growth, results are available promptly. The EAT results were confirmed and it was also found that there are higher substance concentrations in the cytoplasma of Wa-256 cells than in cells in neighboring liver tissue. During the period when substance was being liberated from the tablet, substance concentration in the blood was constant, and considerably lower than in ascites fluid or cytoplasma. This confirms that cysteine-2:1-crotonal adduct can be applied systemically from a subcutaneous depot to a model tumor in a cytostatic concentration.

Inhibition of melanoma B16-F10 growth by lipid peroxidation product 4-hydroxynonenal.

Since a gradual benign-to-malignant progression of murine melanoma B16 after exposure in vitro to hypoxia was described recently, the aim of this study was to test if exposing melanoma B16-F10 cells to aldehyde 4-hydroxynonenal (HNE), which is considered not only as one of the major "second toxic messengers" of oxygen free radicals (or oxidative stress), but as a normal constituent of many cells and tissues, might have opposite effects. Treatment of the tumor cells with 50 microM HNE in vitro or in vivo did not prevent development of the tumors, but inhibited their growth. Tumor growth inhibition was equal for in vitro and in vivo treatment, but appeared after a delay of almost one week, since there was no difference of the tumor volume to the control observed during the initial period of the tumor growth. Similarly, both HNE treatment of the tumor cells before transplantation and HNE treatment of the melanoma bearing mice resulted in equally prolonged survival time. Thus, the results obtained suggest that while hypoxia could increase the malignancy of the murine melanoma cells, exposing these cells to one of the major "second toxic messengers" of oxygen free radicals, HNE, has almost opposite effects and further indicate the possible use of the aldehyde in vivo.