Conventional bladder wash cytology performed by four experts versus quantitative image analysis.

Bladder wash cytology provides superior results for the detection of bladder malignancies than does voided urine analysis. Image analysis systems have been developed for quantification of cytologic features. In this study, routine bladder wash cytology is compared with an automated image analysis system (QUANTICYT). We studied a random set of 100 bladder wash samples from a population of 1614 patients in follow-up after bladder cancer. Four experienced pathologists interpreted the same 100 Papanicolaou-stained slides. Cytologic and image analysis results were compared for prediction of a cystoscopic lesion, histologic abnormalities, and tumor recurrence. After application of receiver operating characteristic curves, prediction of a cystoscopic lesion by cytology and image analysis was comparable. Both the image analysis system and the cytologic examination detected all of the high-grade lesions. Image analysis was superior to cytologic analysis for the prediction of tumor recurrence after normal findings at cystoscopic examination.

Quanticyt: karyometric analysis of bladder washing for patients with superficial bladder cancer.

OBJECTIVES: Quantitative cytology by image-analysis techniques enables objective interpretation of nuclear features in light microscopic images. QUANTICYT, a quantitative karyometric cytology system, was used in the follow-up of patients with superficial bladder cancer. METHODS: From 1992 to 1995, 4137 samples from 1412 patients were obtained. At 1-year follow-up after the initial bladder washing, a tumor recurrence rate of 21% was found. In this period, tumor progression to invasive disease occurred in 1.6% of patients. Scoring of tumor by the QUANTICYT system was based on two nuclear features: the 2c deviation index and the mean of a nuclear shape feature: MPASS. RESULTS: The method was found to be reproducible and superior to visual cytologic interpretation. QUANTICYT analysis of the bladder washings resulted in a score of low, intermediate, and high risk. In a multivariate analysis, highest grade of earlier tumor and QUANTICYT risk score were the best predictors of tumor recurrence and progression. For the easy application of QUANTICYT analysis in daily routine, a report form that included patient history and DNA histogram was developed. CONCLUSIONS: QUANTICYT karyometric analysis of bladder-wash material proved a useful, clinically applicable grading tool in the follow-up of patients with superficial bladder cancer, with sufficient power to be used in decision-making in the individual patient.

Antitumoral effects of liarozole in androgen-dependent and independent R3327-Dunning prostate adenocarcinomas.

We examined the in vivo antitumoral effects of liarozole against androgen-dependent and independent Dunning rat prostatic tumors. Liarozole, applied as a dietary admixture, at a dose of 120 mg./100 gm. food, equivalent to 100 mg./kg. per day, inhibited the growth of the slow growing, well-differentiated, androgen-dependent Dunning-H tumor (median tumor volume decrease of 60%). At the same dose it also significantly reduced the growth of the androgen-independent, moderately differentiated PIF-1 (-60%) and androgen-independent, anaplastic AT-6 tumors (-73%). The growth of AT-6 sq tumor showing squamous metaplasia was unaffected by liarozole. When administered by oral gavage, liarozole at 40 (-82%) mg./kg. twice a day was as effective as castration (-92%) in reducing the androgen-dependent, poorly differentiated Dunning R3327-G tumor. Liarozole, administered by gavage, twice a day, also significantly reduced median tumor volume in the androgen-independent, AT-6 sq (-90% at 60 mg./kg., twice a day). This difference between liarozole administration by gavage and food admixture will have to be taken into account in further experimental studies. Inhibition of the growth of several androgen-dependent and, chiefly, androgen-independent Dunning prostate carcinoma sublines that differ widely in their histological degree of differentiation and growth rate suggests that liarozole may be a suitable agent for evaluation in second line treatment of hormone refractory prostate carcinoma in patients who relapse after androgen ablation.

Antitumor effects of bacillus Calmette-Guerin in a syngeneic rat bladder tumor model system, RBT323.

We have studied the antitumor effects of Bacillus Calmette-Guerin (RIVM strain) in the syngeneic rat bladder tumor model RBT323. In an immunohistochemical infiltrate study we compared the antitumor effects of BCG with the immunopathological findings in order to get more insight into the possible effector mechanisms of BCG. The antitumor effects of BCG appeared not to be dose-dependent, in the dose range tested. In rechallenge experiments no difference in growth of control tumors was seen between rats pretreated- or not pretreated with BCG. There was, however, a significant increase in antitumor effect of BCG after pretreatment with BCG. Immunohistological examination of BCG treated tumors revealed infiltrates consisting of macrophages, B-cells and T-cells. These results imply that whereas no specific response against the RBT323 cells is generated by BCG treatment, possibly BCG induced antigens do serve as immunogens in this nonimmunogenic syngeneic rat bladder tumor model system.

The rat bladder tumor model system RBT resembles phenotypically and cytogenetically human superficial transitional cell carcinoma.

A cohort of 300 ACI rats was kept under standard laboratory conditions. After 30 months or upon natural death, complete autopsy was performed. In the genitourinary tract four kidney and five bladder tumors were found. Two of these bladder tumors, RBT323 and RBT157, are serially transplantable. In the fifth transplant generation the RBT323 tumor becomes metastatic to the lungs in more than 90% of animals. The metastatic ability of the RBT157 tumor changes from low to intermediate (50% of the rats have lung metastases) in the fourth passage. Histologically, the initial passages of the RBT323 and 157 tumors are grade II transitional cell carcinoma (TCC). The histological pattern of the RBT157 tumor remains essentially unchanged, whereas the RBT323 tumor progresses to a grade III tumor in the third passage. Electron microscopical studies reveal oblong elliptical and round vesicles lined by an asymmetrical unit membrane in the tumor cells, which stresses the urothelial origin of the tumors. Immunohistochemically both tumors show expression of cytokeratin 5, 7, 8 and 18. The progression of the tumors to a metastatic phenotype, however, is not associated with a specific change in the morphological characteristics. Cytogenetic analysis shows that both tumors are peridiploid with few marker chromosomes. Interestingly, both of these independently arising tumors exhibit a loss of chromosome 5. Rat chromosome 5 is syntenic to the major portion of human chromosome 9 (p23-qter). Loss of chromosome 9 is a cytogenetic trait of human superficial TCC, hence the RBT model is also in cytogenetic respect similar to human TCC.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of the amount of dietary fat on the development of mammary tumors in BALB/c-MTV mice.

The relationship between dietary fat consumption and the incidence of breast cancer, if any, needs to be quantified so that dietary guidelines can be issued for the prevention of breast cancer. Frequently, only two widely different dietary fat levels, often differing in essential fatty acid content, have been compared in animal models. Moreover, the latent period in common animal models for breast cancer is very short and does not reflect the relatively long latent periods in human breast cancer. We describe a study with BALB/c-MTV mice, a strain with a high average tumor incidence and a latent period of over 60 weeks on average. The mice were fed diets with fat levels ranging from 10% to 40% of energy, in which fat was isocalorically substituted for carbohydrates. The level of linoleic acid in these diets was kept constant at 6.5% of energy. Both the mean tumor incidence and latent periods of the groups fed diets with 10-16% of energy as fat were not significantly different from each other. There were also no differences between these parameters in the groups fed 22-40% of energy as fat. However, the mean incidence and latent period of the groups fed 22% or more of energy as fat was significantly higher than that of the groups fed less fat. We conclude that above about 22% of energy, fat does not influence the incidence and latent period of mammary tumors in BALB/c-MTV mice.

Synergistic antitumor effects of rat gamma-interferon and human tumor necrosis factor alpha against androgen-dependent and -independent rat prostatic tumors.

We have examined the antitumor effects of rat gamma-interferon (IFN-gamma) and human tumor necrosis factor alpha (TNF) against androgen-dependent and -independent Dunning rat prostatic tumors. In vitro studies, using the double layer soft agar assay, showed a very limited antiproliferative activity of the drugs in the dose range tested (1-1000 units IFN-gamma and/or 1-1000 ng TNF/dish). For in vivo studies IFN-gamma and TNF were administered s.c., peritumorally. IFN-gamma was given 3 times/week, 8,000 or 80,000 units/rat, and TNF 5 times/week, 10 or 100 micrograms/rat. IFN-gamma and TNF monotherapy were not significantly effective in inhibiting tumor growth, except for IFN-gamma against the androgen-independent MatLyLu tumor. Combinations of IFN-gamma and TNF had synergistic antiproliferative effects against all four tumor lines tested; however, complete growth inhibitions could not be achieved. Survival studies showed significant increase in survival of tumor-bearing rats.

Differential antiproliferative activities of alpha- and gamma-interferon and tumor necrosis factor alone or in combinations against two prostate cancer xenografts transplanted in nude mice.

We have investigated the antiproliferative effects of recombinant human alpha- and gamma-Interferon (IFN) and recombinant human Tumor Necrosis Factor alpha (TNF) against the hormone-independently growing PC3 and DU145 prostatic tumor lines. Subcutaneous, peritumoral administration of the drugs was started 24 hours after subcutaneous implantation of 1-2 mm3 tumor pieces. IFN was given three times per week and TNF five times per week. IFN-alpha (dose-range 0.5-5 ng/gram bodyweight) had significant growth-inhibiting effects against the PC3 tumor, but showed no significant antitumor effects against the DU145 tumor. IFN-gamma monotherapy (dose-range 8-80 ng/gram bodyweight) was less effective than IFN-alpha. 500 ng/gram TNF produced growth inhibition of both tumors, whereas the lower dose (50 ng/g) was only effective against the PC3 tumor. IFN-alpha and -gamma combination treatment had significant antiproliferative effects against the PC3 tumor, but not against the DU145 tumor. Combinations of IFN-alpha and TNF were very effective against both xenografts; some combinations resulted in complete growth inhibition. IFN-gamma and TNF combinations also showed significant antitumor effects against both tumor lines. We therefore conclude that cytokine combination treatment may provide a new approach in the treatment of hormone-escaped prostatic tumors.

Combined effects of tumor necrosis factor alpha and radiation in the treatment of renal cell carcinoma grown as radia spheroids.

We have investigated the antiproliferative effects of Tumor Necrosis Factor Alpha (TNF) and radiation on a recently described rat renal cell tumor line grown as multicellular tumor spheroids (MTS). Treatment commenced when the spheroids had reached a diameter of 250 microns. TNF was diluted in the tissue culture medium in different concentrations, ranging from 250-1000 ng/ml. TNF monotherapy had a dose-dependent inhibiting effect on spheroid growth. Single-dose irradiation with 2, 4 or 6 Gy also retarded spheroids significantly in their growth. In the combination treatment the highest dose of TNF (1000 ng/ml) was added 4 hours prior to radiation. TNF could not induce a potentiation of the radiation injury at 2 Gy. The combination with 4 Gy, however, had additive and the combination with 6 Gy synergistic antiproliferative effects; in these treatment regimens respectively 2 and 5 out of 24 spheroids were controlled, i.e. cured. These experiments suggest that TNF in combination with radiotherapy may be beneficial for the treatment of renal cell carcinoma or cancer in general.

Effects of meal size reduction on postprandial variables in male volunteers.

To investigate the effects of meal size reduction on postprandial variables, adult male volunteers consumed at noon hot lunches composed of cooked white rice, fried chicken fillet, raisins, bigarreaus and curry sauce with carbohydrate/fat ratios of either 0.77 or 2.04 (37-57 en% carbohydrate, respectively). Prior to the actual experiment, the individual meal sizes were determined by allowing the volunteers to eat a self-preferred amount of the high carbohydrate meal. The test meals were adjusted to either 70 or 100% of the energy content of this individually chosen meal size. A higher degree of satiety was detected up to 4 h after the 100%-size carbohydrate-rich meal as compared to that after the fat-rich meals and the 70%-size carbohydrate-rich meal. The reduction in meal size had only small effects on the postprandial curves of glucose, insulin, free fatty acid and total glycerol in the blood relative to those induced by the variation in carbohydrate/fat ratio in the meals. No significant effects of meal size were detected on the postprandial curve of free glycerol. Thus limited restriction in the size of a meal relative to the ad libitum intake of an individual may be expected to lead to only minor effects on the various postprandial curves in meal-eating studies.

Minor difference in postprandial responses of men between starch and sugar when replacing fat in a normal meal.

Healthy male volunteers consumed hot lunches consisting of cooked white rice, fried chicken fillet, raisins, bigarreaus (sweet cherries) and curry sauce with carbohydrate: fat ratios of either 0.77 or 2.04, and polysaccharide: (mono + di)saccharide ratios (P:MD) varying between 5.3 and 0.95. Before and at various time intervals after the start of the meal, blood samples were analysed for glucose, insulin, triacylglycerol (TG), free glycerol (FG), free fatty acids (FFA) and cholesterol. Elevated postprandial glucose and insulin peaks were induced by the meals containing a larger amount of carbohydrate. The type of carbohydrate in the meal appeared to have little or no effect on the peak maximum. A small second glucose peak was seen at 2 h after the meals with P: MD greater than 3.1. The TG concentration in the blood showed a similar and rapid rise. After the meal containing the largest amount of simple sugars, the TG curve levelled off 1 h after the start of the meal and then remained at a nearly constant level (1.5 mM). In contrast, a larger amount of complex carbohydrates induced a TG concentration which rapidly rose to a maximum of 1.75 mM, and subsequently decreased slowly to somewhat below that of the simple sugar-rich meal at 4 h. The postprandial curves after the fat-rich meals showed a continuous rise of TG up to a maximum of about 2 mM at 2 h, and a subsequent slow decrease. FG and FFA all showed a rapid drop immediately after the start of the consumption of the meals, followed by a rebound at 1 h. The postprandial curves of total cholesterol, as well as the areas below the curve of both total cholesterol and free, unesterified cholesterol were lower after the carbohydrate-rich meals than after the fat-rich meals. This is attributed to the larger amount of cholesterol in the fat-rich meals.

Effects of varying the carbohydrate:fat ratio in a hot lunch on postprandial variables in male volunteers.

1. Healthy male volunteers consumed at noon, hot test meals with four different carbohydrate:fat ratios varying between 2.64 and 0.50, and composed of fried beefsteak, mashed potatoes, French beans, and a dessert of custard with mashed peaches. The energy content of the meals was 40% of the daily intake of the volunteers, estimated from their individual dietary histories. 2. Before, and at different times after the start of the meal, blood samples were taken and a number of indices of carbohydrate and lipid metabolism were determined in the samples, i.e. glucose, insulin, free fatty acid, free and total glycerol, free and total cholesterol, high-density-lipoprotein (HDL)-cholesterol and low-density-lipoprotein (LDL)-cholesterol. 3. Increasing the carbohydrate:fat ratio resulted in higher postprandial peaks of glucose and insulin. In addition, the peak area under the postprandial glucose curve showed a significant increase. The peak area under the postprandial insulin curve had also increased, indicating that a larger amount of insulin was secreted by the pancreas on increasing the carbohydrate content in the meal. There was no significant correlation between the height of the postprandial peak of blood glucose and the size of the meal. 4. All four meals caused elevated postprandial blood triacylglycerol levels. However, the decline of this elevated level took a much longer time after the meals with the lower carbohydrate:fat ratios, i.e. containing larger amounts of triacylglycerols. There was a significant decreasing linear relation between the carbohydrate content of the meals and the peak area under the postprandial triacylglycerol curve. Free glycerol and free fatty acids showed lower postprandial levels in the blood after the meals with the higher carbohydrate:fat ratios, and the peak areas of the postprandial curves of both variables displayed a significant decrease. Little or no effect of the meal carbohydrate:fat ratio was observed on the postprandial concentrations of total cholesterol, unesterified cholesterol, HDL-cholesterol or LDL-cholesterol.

The effect of dietary carbohydrate:fat ratio on energy intake by adult women.

The effect of the dietary carbohydrate:fat (C:F) ratio on the spontaneous energy intake by healthy adults was investigated by comparing a high-carbohydrate diet (fat 24%, carbohydrate 58%, protein 18% of energy) and a high-fat diet (fat 47%, carbohydrate 35%, protein 18% of energy) in a 2 X 2 week cross-over design. Subjects were 22 healthy nuns in a Trappist convent with very regular activities. The diets consisted of combinations of liquid formula (75%) and standardized snacks (25%). The difference in C:F ratio was concealed: energy density, taste and appearance were similar. Energy consumption was recorded continuously. The mean daily energy intakes remained constant: 8276 kJ (1978 kcal). The difference in mean daily energy intake between diets was 73 kJ +/- 180 (SEM). Small changes in body weight were observed, but these are argued not to indicate definitive effects. It is concluded that changing the C:F ratio within commonly occurring ranges does not influence the spontaneous energy intake of healthy adults. The composition of the dietary fat was kept constant. Under practical conditions a change in the C:F ratio will also induce a change in the fatty acid composition of the diet, which might affect the energy intake regulation. Other experiments are required to see whether the C:F ratio can affect body composition or other physiological parameters in the long run.

[Physiologic effect of a dietary regimen rich in soy proteins in man]. / Effets physiologiques d'un régime alimentaire riche en protéines de soja chez l'homme.

The increasing availability of palatable soya bean protein materials for use in human foods raises the question of possible effects on the health and well-being of the consumer. On the basis of available evidence, no unfavourable effects are to be expected, but apart from nitrogen balance investigations, systematic human experiments with these novel foods are scarce. Therefore, we carried out a large-scale experiment covering many physiological and health aspects. Two diets were compared in 4 + 4 weeks cross-over design with 92 healthy volunteers. One diet contained a wide variety of soya protein foods (test diet), about 25% of the protein intake being from soya, the other (control) diet contained similar products made from conventional protein sources. The diets were given in two identical menu cycles. Blood, urine and feces were sampled at the end of each period. Health status and subjective reactions were monitored throughout. About 90 out of the more than 100 parameters investigated did not show any difference. Statistically significant reactions to the diet composition were found in the following areas: of the blood serum enzymes, alkaline phosphatase was higher after consumption of the test diet, while serum inorganic phosphate showed a decrease. The higher magnesium content of the soya protein materials was reflected by an increase in fecal excretion, and in serum levels in the fed state. Fasting serum levels, however, were lower on the soya diet. Measurement of the intestinal noise indicated that more intestinal gas was produced on the test diet; this was confirmed by reports from the volunteers. Immunological tests showed that the females had increased IgA and soya specific IgE levels on the test diet, but no indications for allergenicity were found. All the mentioned effects were well within normal physiological ranges and do not indicate unfavourable trends. We conclude, that these results confirm the prevailing view that soya bean protein materials are acceptable ingredients for our daily food.