Growth of a spontaneous canine prostatic adenocarcinoma in vivo and in vitro: isolation and characterization of a neoplastic prostatic epithelial cell line, CPA 1.

Neoplastic epithelium derived from a spontaneous canine prostatic adenocarcinoma has been maintained and grown in cell culture and as xenografts in athymic mice. An epithelial cell line (CPA 1) has been isolated from primary cultures and has been partially characterized in vitro. The growth of this cell line was not modified by either androgens or estrogens, and high-affinity receptors for these steroids could not be demonstrated in these cells. Xenografts were serially transplantable, with growth being similar in both sexes. Receptors for androgens and estrogens could not be detected in homogenates of xenografts or primary tumor. The histological appearances of serially transplanted tumors, and of xenografts generated by inoculation of the cell line (CPA 1) and several cloned substrains, were very similar to that of the primary tumor and were judged to be well differentiated. The characteristics of this neoplastic cell type have been compared with those of normal prostatic epithelium.

Steroid hormone concentrations in relation to patient prognosis and prostate tumour grade.

Survival of patients who received endocrine therapy as first-line treatment for their prostatic cancer was statistically analysed in relation to several parameters, primary tumour stage, metastatic status, age, pretreatment plasma hormone concentrations and Gleason grade. Prognostic indices were derived for both M0 and M1 patients in which Gleason grade and plasma testosterone concentrations were significant prognostic factors. In the M1 patients growth hormone values were also significant and to a lesser degree age. The relationship of Gleason grade to testosterone, growth hormone, prolactin, the gonadotrophins and age was also analysed. No significant differences in any of these hormones was noted with increasing Gleason grade but the age of patients with Grade 5 tumours was significantly lower.

Cortisol concentrations in the umbilical artery and vein of breech-presenting infants at term in relation to the method of delivery.

Umbilical cord artery and vein cortisol levels and cord artery pH were measured in 32 breech deliveries. There was no detectable increase in fetal cortisol output in relation to fetal acidosis. It is considered that elevation of fetal cortisol levels is caused by maternal transfer of this hormone transplacentally during stressful delivery rather than by enhanced fetal adrenal activity.

A prognostic index for the clinical management of patients with advanced prostatic cancer: a British Prostate Study Group investigation.

Patients with histologically proven carcinoma of the prostate (n = 186) were initially assessed and followed up according to the standardized protocol of the British Prostate Study Group, urologists from which contributed patients to this investigation. These patients were given either endocrine therapy or orchidectomy as first line treatment; the ratio of the number of patients receiving these two treatments was similar in each group of subjects compared for survival. Prognostic indices were derived for all patients and for those classified according to the presence (M1) or absence (M0) of metastases. The prognostic indices were derived from clinical and hormone data obtained at initial presentation. Whereas the degree of tumor differentiation and plasma testosterone concentrations were significant prognostic factors in both M0 and M1 disease, growth hormone was only significant in M1 patients, where age was also of borderline significance; elevated growth hormone, higher Gleason grade, younger age, and lower testosterone indicated a poorer prognosis in M1 patients. These findings indicated the feasibility of selecting a poor prognostic group of patients that may derive benefit from a more aggressive therapy.

A study of the androgenic function of the epididymis.

Rats were subjected to bilateral orchidectomy or orchido - epididymidectomy and maintained on either 500 micrograms testosterone or testosterone propionate daily. The ventral lobes of the prostate were subsequently excised and examined for androgen receptors in terms of the total present in the cytosol and the nucleus, the proportion unoccupied by endogenous androgen and the relative populations that were nuclease excisable or nuclease resistant in the two groups of animals. A further group of animals was subjected to unilateral deferential venotomy and the same parameters examined in the ipsi- and contra-lateral lobes of the ventral prostate and the seminal vesicles. In the absence of the epididymides there was a reduction in the number of receptors per prostatic cell and an increase in the proportion that were unoccupied. The nuclei from these glands contained fewer receptors than did those from the animals in which the epididymides had not been excised. The effect of unilateral deferential venotomy was to bring about a relative increase in the number of cytoplasmic receptors in the ipsilateral lobes of the ventral prostate with a much greater proportion unoccupied compared with the lobes contra-lateral to the ligation. There was again an increase in the proportion of nuclease-sensitive receptors in the nuclei ipsilaterally. The conclusions are that the absence of the epididymides in androgen-maintained rats or deferential venotomy induces a relative androgen- deficiency of the prostate and seminal vesicles as reflected in the androgen receptor populations of these organs.

Carcinoma of the prostate: relationship of pretreatment hormone levels to survival.

Pretreatment hormone levels were determined in 222 patients with prostatic cancer and their prognostic value assessed. The patients were grouped into yearly survival categories and only those whose cause of death was due to the disease were included in the study. Low concentrations of testosterone in plasma at the time of diagnosis related to a poor prognosis. Patients who died within 1 yr of diagnosis had the lowest mean plasma levels of this steroid. The pretreatment mean plasma testosterone concentrations were found to be higher as the survival period of the various groups lengthened. This relationship was observed both when the total data were analysed and also when the patients were subgrouped depending on clinical evidence of spread of the tumour beyond the prostatic capsule (T3) or on the presence of metastases (M1). High pretreatment plasma concentrations of luteinizing hormone were also associated with poor survival. Follicle-stimulating hormone, prolactin and growth hormone concentrations did not correlate with survival time. The indications from this study are that poor testicular function is associated with early death from prostatic carcinoma and that the measurement of blood levels of testosterone at diagnosis could provide a prognosis of subsequent life-span.

Growth and histology of four canine mammary tumour lines established in nude mice.

A colony of nude (BALB/c/Nu/Nu) mice has been established and used to produce and study transplantable tumour lines derived from spontaneously occurring canine mammary neoplasms. Four primary tumours of differing morphological types were studied. Two were adenocarcinomas (MS306 and PD6014) with varying degrees of differentiation, the third (F5010) was a complex adenocarcinoma composed of both stromal and epithelial elements and the fourth (V5500) was a fibrosarcoma. Tumour fragments were implanted into the subcutaneous tissue of 4- to 6-week-old mice and resulted in tumour growth in each case. There was a latent period of 1 month for tumours PD6014, F5010 and V5500 and of 3 months for tumour MS306 before tumour growth ensured. After serial transplantation this period decreased to 1-2 weeks for the first 3 tumours and to 6 weeks for the last. After the initial lag period, tumour volume increased logarithmically in all cases with doubling times of 6-20 days. Each tumour line has been passaged through 3 serial transplantations with 3 of the tumours retaining their original histological appearance whilst the fourth became slightly more dedifferentiated.

Initial characterisation of oestrogen receptors in canine mammary tumour lines maintained in nude mice.

Proteins binding oestradiol with high affinity and low capacity have been characterised and quantitated in 4 newly established canine mammary tumour lines, MS306, V5500, F5010 and PD6014, maintained in nude mice. These receptor proteins, specific for oestrogens, were found in all the tumour lines, both in initial implants and all subsequent passages. Receptors had equilibrium dissociation constants for oestradiol in the range of 33-210 pM, and sedimentation coefficients of 4 S and 8 S.

Cytoplasmic steroid effects on nuclear RNA polymerase activity in canine mammary carcinomas.

DNA-dependent RNA polymerase activity has been studied in isolated nuclei from canine mammary tumours. Initial experiments showed high levels of RNase activity in this tissue; accordingly, routine assays were terminated before loss of acid-precipitable radioactivity was evident. RNA polymerase A and B activity in isolated nuclei were shown to be increased by addition of receptor-containing cytosol previously incubated with oestradiol-17 beta, dihydrotestosterone or R5020. Where no receptor was present, as measured by saturation binding assays and sucrose density gradient analysis, there was no corresponding increase in polymerase activity. The steroid antagonists tamoxifen and cyproterone did not elicit any response even when their corresponding steroids produced a 1- to 2-fold stimulation of polymerase activity. Steroid-induced effects proved to be dose-dependent, with half maximal responses for oestradiol-17 beta 8 X 10(-8)M, R5020 2 X 10(-6)M and dihydrotestosterone 9 X 10(-6)M.

Prolactin and breast skin temperature rhythms in postmenopausal women with primary breast cancer.

Plasma prolactin and cortisol were determined and breast skin temperatures measured in 8 postmenopausal diurnally active-nocturnally resting women, 7 with primary breast cancer and 1 with benign breast disease. In all subjects, prolactin peaked during the early morning hours. Cortisol and oral temperature served as internal physiological markers of anticipated rhythmicity in adrenocortical function and metabolism. In all patients studied, the tumour site had an increased mesor (midline-estimating statistic of rhythm) and reduced amplitude of the circadian breast skin temperature rhythm when compared to a similar site on the contralateral breast. Thermal asymmetry was also observed between left and right breasts. A cancer, if in the left breast, may reinforce lower amplitude and perhaps a higher mesor (associated with the healthy left breast in these particular patients), whereas if it is located in the right breast, it tends to nullify inter-mammary differences in mesor and amplitude. Screening for breast cancer appears to require subjects being monitored on at least two occasions before thermal abnormalities due to tumour or preneoplasia become discernible.

An examination of blood steroid and gonadotropin concentrations in relation to fertility status and testicular function in men.

Plasma concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin (PRL), testosterone (T), and 17 beta-estradiol (E2) have been measured in men complaining of infertility in comparison with men of proven fertility. Subgrouping of patients was achieved on the basis of the presence or absence of sperm in the ejaculate and further by the concentration of sperm or by testicular score. The levels of plasma LH, FSH, PRL, and T were found to be significantly different in the fertile men, compared with both infertile men with sperm in their ejaculates and azoospermic men. There were no significant differences between the groups for E2. There appeared to be an inverse relationship between LH concentrations and sperm count in both fertile and infertile men. FSH levels did not vary significantly in the fertile men in relation to sperm count grouping but were significantly less than those found for the infertile men with sperm. Azoospermic patients with high testicular scores had FSH levels indistinguishable from those of the fertile men. The results are discussed in terms of testicular abnormalities and on the interrelationship between the hormones examined.

PIP: Plasma concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin (PRL), testosterone (T), and 17beta estradiol (E2) have been measured in men complaining of infertility in comparison with men with proven fertility. Subgrouping of patients was achieved on the basis of the presence or absence of sperm in the ejaculate and further by the concentration of sperm of by testicular score. The levels of plasma LH, FSH, PRL, and T were found to be significantly different in the fertile men, compared with both infertile men with sperm in their ejaculates and azoospermic men. There were no significant differences between groups for E2. There appeared to be an inverse relationship between LH concentrations and sperm count in both fertile and infertile men. FSH levels did not vary significantly in the fertile men in relation to sperm grouping count but were significantly less than those found for infertile men with sperm. Azoospermic men with high testicular scores had FSH levels indistinguishable which were from those of fertile men. The results are discussed in terms of testicular abnormalities and the interrelationship between the hormones examined.

Epithelial and fibroblastoid cell lines derived from the normal canine prostate. I. Separation and characterization of epithelial and stromal components.

Cell cultures displaying exclusively epithelial or fibroblastoid morphology have been isolated by spillage and collagenase digest techniques, respectively. Primary cultures of both cell types have been readily subcultured. The use of a type I collagen substrate has been shown to be essential to the growth of normal prostatic epithelium in monolayer cultures. The ability to generate replicate subcultures of both cell types has allowed the quantitative characterization of the mitogenicity of fetal bovine serum and insulin in early subcultures. The control of culture conditions has permitted uniform cell population growth in early subculture with regular population doubling times in log phase of growth. Epithelial cultures have been shown to display many ultrastructural characteristics common to the normal epithelium of the canine prostate.

The effect of the removal of the various accessory sex glands on the fertility of male rats.

Accessory sex glands were removed from rats which were then allowed to mate with females. Ablation of the ventral prostatic lobes appeared to have no affect on the animals' fertility whereas removal of the dorsolateral lobes or of the seminal vesicles induced complete infertility. Excision of the coagulating glands caused infertility in 12/16 males.

Characterisation of breast skin temperature rhythms of women in relation to menstrual status.

Circadian breast skin temperature rhythms were characterised throughout the menstrual cycle, for various locations on the left breast of ambulatory women. All subjects exhibited highly significant circadian rhythms (P less than 0.001). Changes in rhythm parameters, such as the mesor, amplitude and acrophase, were observed during the menstrual cycle. No consistent trend in these rhythm parameters was observed between subjects in relation to menstrual cycle stage. Experimental and statistical techniques used to characterise circadian rhythms in pre-menopausal women were applied to a post-menopausal woman with primary breast cancer. Comparison of rhythm parameters associated with the tumour area and corresponding site on the contralateral breast showed abnormal thermal characteristics such as elevated mesor values, decreased amplitude as well as changes in the timing of the acrophase. These properties may be exploited for the early detection of breast cancer. The project also involved the design and testing of an ambulatory device, known as the 'chronobra', for the measurement of breast skin temperature. The performance of the chronobra was in close agreement with reliable, conventional equipment. The chronobra now allows studies of breast skin temperature rhythms associated with breast disease to be extended.

Androgen steroid-receptor binding in the canine epididymis.

The epididymis of the dog has been studied with regard to its ability to bind androgens with a high affinity and low capacity. A proteinaceous molecule was demonstrated in the high-speed supernatant fraction of disrupted cells that bound DHT and sedimented in a sucrose density gradient with a sedimentation coefficient of 8S with respect to bovine serum albumin. This binding protein was able to transport DHT to the nucleus at elevated temperatures where it sedimented as a 4S in a 5-20% sucrose density gradient. Even though this protein appears to be similar to other androgen receptors in other accessory sex organs in a variety of mammalian tissue, it is different from androgen-binding protein (ABP). The apparent specificity of the receptor protein for androgens was demonstrated using competition studies. The molecular configuration of the steroids was found to be instrumented in this selectivity. A reduced affinity of the androgen receptor was found when the steroid did not possess a hydroxyl group at the 17 beta-position and a 3-oxo-group irrespective of the degree of saturation of the A-ring. Studies concerning the physical properties of the androgen binder revealed a protein with a molecular weight of 220,000, a Stokes' radius of 54A, and a frictional ratio of 1.35.