Increased levels of fibroblast growth factor-like activity in urine from patients with bladder or kidney cancer.

Growth factor activity was partially purified from human renal tumors and a human bladder cancer cell line by heparin-Sepharose chromatography. This activity stimulated bovine capillary endothelial cell proliferation and DNA synthesis in BALB/c 3T3 cells. Partially purified growth factor preparations from these tumors contained a protein with an approximate molecular weight of 17,000 which was recognized by a polyclonal antiserum raised against a peptide fragment of basic fibroblast growth factor (FGF). This growth factor activity appears to be related to basic fibroblast growth factor. Measurement of FGF-like activity in 50 urine samples from 32 adult males showed that 55% (6 of 11) of the urine samples from patients with bladder cancer and 100% (7 of 7) of the urine samples from patients with kidney cancer contained activity equivalent to more than 20 ng of basic FGF/h of urine production. In contrast, only 6% (2 of 32) of the urine samples from controls, patients with a benign disease, or patients with a history of bladder or kidney cancer contained this level of growth factor activity. These results suggest that patients with bladder or kidney cancer release an FGF-like factor into urine which may be used as a marker for these tumors.

Glucocorticoid receptors in depression: relationship to the dexamethasone suppression test.

Cytoplasmic glucocorticoid receptor content wa quantitated in lymphocytes from unmedicated depressed patients and control subjects before and after a standardized dexamethasone suppression test. Depressed patients (N = 11) had significantly lower (32%) basal cytoplasmic glucocorticoid receptor content than the control group (N = 14). Suppression of serum cortisol (5.0 micrograms/dl or less) in both control and depressed subjects (N = 16) following dexamethasone (1 mg) was associated with a decrease in lymphocyte cytoplasmic glucocorticoid receptor number, whereas no such change occurred in cortisol nonsuppressors (N = 9). Changes in receptor concentration were positively correlated with postdexamethasone serum cortisol levels and with the inhibitory effect of dexamethasone on mitogen-induced lymphocyte proliferation.

Age, sex and gonadal influences on oestrogen receptor content in the rat liver.

Liver cytosol oestrogen receptor (OR) content of intact and gonadectomized male and female rats of defined age (1-18 months) was measured by the controlled pore glass bead assay. The OR content of animals of both sexes aged 3 weeks was low to insignificant (0.12 +/- 0.04 (S.E.M.) pmol/g wet wt liver). In the intact animal, liver OR reached maximum levels at an earlier age in the male (1.35 +/- 0.12 pmol/g at 28-32 weeks) than in the female (4.89 +/- 0.17 pmol/g at 44-52 weeks). In the mature female the OR is thus maintained at a level threefold greater than in males of comparable age. Levels of OR in the gonadectomized male tended to be higher, at all ages, than those in the intact male but this difference was not statistically significant. Ovariectomy was associated with a decreased OR content in rats aged less than 6 months and an increase in the older rats, but these effects (of ovariectomy) were not statistically significant. In the females, regression analysis of pooled data, combining all ages, indicated a significant positive correlation between OR and liver wet weight, independent of gonadal status; no such relationship was found with the males. The results indicate that factors influencing liver cytosol OR content include maturity, age and sex whereas the gonads have little obvious effect. These findings lend further support to the concept that sex differences exist in the liver.

A controlled pore glass bead assay for the measurement of cytoplasmic and nuclear glucocorticoid receptors.

An assay for the quantitation of cytoplasmic and nuclear glucocorticoid receptors in lymphoid tissue has been developed using controlled pore glass (CPG) beads. Soluble receptor--3H-steroid complex (cytosol or nuclear extract) is adsorbed quantitatively within the crevasses of porous glass beads. Excess labeled steroid as well as most non-specifically bound steroid is easily washed away, leaving the hormone-receptor complex retained by the beads. Bound 3H-steroid is eluted with ethanol and measured for radioactivity. This procedure which is simple, rapid, and highly reproducible is carried out using frozen samples (stable for many months) containing as few as 1 X 10(7) cells. A comparison of the CPG assay to dextran coated charcoal and a whole cell assay demonstrates that CPG and dextran coated charcoal give equivalent measurements of cytosolic receptor concentration, while the CPG and whole cell assays provide equivalent values for total receptor content.

Sulfhydryl sensitivity and [125I]-16 alpha-IODO-17 beta-estradiol binding of estrogen receptor in ovarian epithelial carcinomas.

We find that estrophilin from either the cytosolic or nuclear fractions of ovarian epithelial carcinomas (OVCA) binds irreversibly to controlled-pore glass beads (CPG). The CPG-adsorbed estrophilin releases [3H]-estradiol at 4 degrees C in the presence of 1.25 mM AgNO3; estradiol binding capacity from the nuclear fraction is restored by 10 mM dithiothreitol. The number of available estradiol binding sites in cytosol that are sensitive to AgNO3 correlate with (a) the number of estradiol-inhibitable binding sites found in the 8S region in low-salt sucrose gradients (r = 0.9) but not with (b) estradiol-inhibitable binding in the 4S region (r = 0.3) and thus, as expected, only poorly with (c) estimates of available cytoplasmic estrogen binding sites using a standard dextran-coated charcoal analysis (r = 0.7). Total estrogen binding extracted from the nuclear fraction with 0.5 M KCl and adsorbed to hydroxylapatite agreed with the sulfhydryl blocking reagent sensitive moiety (r = 0.9). Estrophilin from OVCA cytosol or nuclear fractions bound [125I]-16 alpha-iodo-estradiol indistinguishably from [3H]-estradiol. The two forms of radiolabelled-estradiol yielded equivalent data that demonstrated a shift in the estrogen binding moiety from the 8S region to the 4-5S region when 0.5 M KCl was added to gradient analyses of OVCA cytosols. From these observations we conclude that the estrophilin found in OVCA is similar to that found in normal and cancerous tissues of other female reproductive organs and that this estrophilin can bind a biologically active radiohalogenated estrogen potentially useful for imaging or treating these tumors.

Isolation of lung alveolar walls.

Disintegration of peripheral lung tissue followed by filtration through an 80 mesh stainless steel screen yields a filtrate containing tissue particles with properties consistent with their identification as fragments of alveolar walls. Application of this procedure readily yields large quantities of alveolar wall tissue.