Occurrence and de novo biosynthesis of follicle stimulating hormone (FSH) in benign and malignant conditions of human breast.

We report the occurrence as well as biosynthesis of a pituitary hormone, follicle stimulating hormone (FSH) in human breast. Using immunoperoxidase localization technique, both FSH and beta-FSH were localized in cytoplasm of epithelial cells but not in stromal cells. Immunostaining was more intense in benign and malignant specimens as compared to normal. In vitro radiolabelled precursor experiments with breast tissue explants indicate de novo synthesis of FSH. Human milk had higher concentrations of FSH as compared to serum. In gonads, FSH is involved in the cellular growth, differentiation and function. The presence of higher levels of FSH in benign mammary tumors and breast cancer when compared to normal breast supports the suggestion that FSH might have a role in the process of breast malignant transformation.

Differential distribution of inhibin in the anatomical regions of monkey stomach.

Inhibin, a 10.7 kD FSH (follicle-stimulating hormone) suppressing prostatic peptide has been shown to be synthesized and localized in stomach specimen of monkey. In vitro incorporation of labelled amino acid (3H-leucine) into inhibin followed by specific immunoprecipitation by antiserum to inhibin demonstrated an in vitro de novo biosynthesis of inhibin by monkey stomach. Moreover, the synthesis of inhibin was found to be maximum in fundic zone of gastric mucosa compared to cardiac and antral zone. This was supported by immunohistochemical study of three anatomically different regions, especially wherein fundic zone showed intense positive staining for inhibin. Furthermore, the above data was supplemented by quantitative study of tissue inhibin content by RIA which revealed that the fundic zone of gastric mucosa has a much higher concentration of inhibin than cardiac and antral region. The relationship of zonal concentration of inhibin to gastric anatomy appears to be a noteworthy observation and may serve as an useful tool in our understanding of gastric metabolism and activity.

Immunoperoxidase localization and denovo biosynthesis of a 10.5-kDa inhibin in benign and malignant conditions of human breast.

Using immunocytochemistry, we report the occurrence of a 10.5-kDa inhibin in human breast tissue specimens obtained from normal, fibroadenoma and adenocarcinoma cases. The immunostaining for inhibin was confined to the cytoplasm of the epithelium and myoepithelium cells. Expression of inhibin increased in following order: normal (1+); adenocarcinoma and lobular carcinoma in situ (2+) and fibroadenoma (4+). Breast explants has the ability of denovo biosynthesis of inhibin in vitro. In view of the growth modulating regulatory properties of 10.5 kDa inhibin, our findings are suggestive of the potential role of inhibin in breast pathology.

Suppression of DNA synthesis and induction of apoptosis in rat prostate by human seminal plasma inhibin (HSPI).

In vivo administration of HSPI (10.7 kDa FSH suppressing peptide of prostate) to intact adult male rats was found to suppress the basal levels of incorporation of 3H-thymidine into DNA of ventral and dorsolateral lobes of the prostate, in a dose-dependent manner. Interestingly, microscopic examination of the prostate histology of HSPI treated rats revealed a significantly increased incidence of apoptotic bodies which are indicative of cell death. In another experiment, HSPI was also found to suppress the active DNA synthesis in testosterone-induced regrowth of prostate in castrated rats. Thus HSPI can suppress the basal and stimulated DNA synthesis and also induce apoptotic cell death in rat prostate.

Dual regulatory action of prostatic inhibin (10.7 kDa) on DNA synthesis.

Present studies deal with the role of inhibin in proliferation and growth. The effect of inhibin on incorporation of 3H-thymidine in prostatic DNA in vivo as well as by NRK-49F and Balb/c3T3 cell lines in vitro, was investigated. Also studied the immunocytochemical localization of inhibin in normally proliferating and differentiated tissues of human prostate and endometrium. The in vivo studies revealed a suppression of 3H-thymidine uptake both in ventral (33%) and dorsolateral (26%) lobes of rat prostate. Interestingly, the histology of inhibin treated rat prostate manifested amidst the epithelial lining, an appearance of apoptotic bodies which are considered to be indicative of cell death. Further, the immunocytochemical studies for localization of inhibin showed intense staining in the differentiated human prostate and endometrium as compared to the respective proliferative tissues. Is inhibin kept suppressed in these proliferating tissues, because it is antiproliferative? The present in vitro experiments demonstrated that, at low inhibin concentrations, the incorporation of 3H-thymidine is stimulated while at higher doses it is suppressed. Thus, it is clear that prostatic inhibin seems to have a concentration-dependent dual role in the regulation of DNA synthesis.

De novo biosynthesis and localization of immunoreactive inhibin (10.7 kDa) in normal human stomach.

We have previously reported the occurrence of inhibin-like peptide in gastric juice of normal men. In the present investigation, normal gastric mucosa was shown to synthesize inhibin, in vitro, as measured by 3H-leucine incorporation (Maximum at 18 h). Furthermore, the immunohistochemical localization studies demonstrated its presence in the acid secreting parietal cells and basal region of foveolar epithelium of gastric mucosa. Surprisingly, the protein secreting zymogen cells remained unstained.

Inhibin (10.7 kD prostatic peptide) in normal, hyperplastic, and malignant human endometria: an immunohistochemical study.

The expression of inhibin, a 10.7 kD follicle-stimulating hormone (FSH)-suppressing prostatic peptide of 94 amino acids, was investigated in normal human endometrium, endometrial hyperplasia, and adenocarcinoma, employing the avidin-biotin immunoperoxidase technique. The antiserum used was raised in rabbits against prostatic inhibin isolated from human seminal plasma. The study included 15 well differentiated, 32 moderately differentiated, and 21 poorly differentiated endometrial adenocarcinomas; 26 simple, five complex, and two complex atypical endometrial hyperplasias; and, for comparison, 25 normal proliferative and 30 normal secretory endometria. In malignant and hyperplastic endometrial tissues, inhibin was localized in the epithelial cytoplasm of endometrial glands while the stroma showed weak reactivity. On the other hand, inhibin was undetectable in the early proliferative phase, but was present on the luminal border of the glandular epithelium in the mid- and late proliferative phases. Secretory endometrium displayed strong inhibin reactivity in the cytoplasm of glandular epithelium and in the stroma. The increased inhibin reactivity in secretory endometrium as compared with the proliferative phase is indicative of a functional role for inhibin in the uterus. In addition, its localization in proliferative, hyperplastic, and malignant endometria suggests a possible regulatory role for inhibin in endometrial proliferation and growth.

Antiproliferative action of prostatic inhibin on NRK-49F and BALB/c 3T3 cell lines.

Prostatic inhibin purified from human seminal plasma (10.7 kDa, 94 amino acids) is very well known for its endocrine action on pituitary to suppress synthesis and secretion of FSH. In the present report we have revealed its antiproliferative action on two fibroblast cell lines, NRK-49F (ED50 = 2.5 ng/ml) and Balb/c 3T3 (ED50 = 24.5 ng/ml) which may mark its emergence as a negative growth regulator.

Inhibition of PHA-stimulated proliferation of peripheral blood mononuclear cells by human seminal plasma inhibin (HSPI).

This report deals with the effect of purified HSPI (10.7 kDa single chain prostatic peptide of 94 amino acids) on PHA-stimulated proliferation of peripheral blood mononuclear cells (PBMNC). HSPI was found to inhibit the incorporation of 3H-thymidine by PHA-stimulated PBMNC in a dose-dependent manner, albeit with the requisite of pretreatment of cells with HSPI. A maximum inhibition was observed on pretreatment for 30 min. These studies thus indicate that HSPI may be interfering with the very early events of PHA-induced signal transduction process in PBMNC.

In vivo suppression of pituitary and circulatory follicle stimulating hormone by human seminal plasma inhibin.

Human seminal plasma inhibin (HSPI), of prostatic origin, has been shown to bring about a dose dependent suppression in pituitary and circulatory FSH concentrations in intact rats. No significant changes in LH levels either in pituitary or in circulation were observed at the doses used. This has further been substantiated by an immunocytochemical staining. A marked reduction in staining intensity for FSH was observed in the pituitary of inhibin treated rats as compared to the controls. None of the purified inhibin peptides from other sources have so far been reported to act on pituitary FSH in vivo. This study thus, for the first time demonstrates an in vivo effect of inhibin (HSPI) on pituitary FSH concentration and secretion.

Immunocytochemical localization of follicle stimulating hormone in normal human stomach.

Immunoreactive follicle-stimulating hormone (IR-FSH) is detected in sections of formalin-fixed and paraffin-embedded gastric mucosal tissue of normal men, using the immunoperoxidase staining technique and specific antisera to hFSH (NIDDK, NIH). Positive staining for IR-FSH was detected in the parietal cells lining the gastric glands of the intermediate zone. The staining was intracytoplasmic and distributed throughout the cytoplasm. IR-FSH was also found to be present in the basal part of the foveolar epithelium. Stromal tissue and nuclei were devoid of the stain. The zymogen cells in the deeper region of the mucosa did not show any detectable staining for IR-FSH. The presence of IR-FSH in gastric mucosa was also detected by radioimmunoassay. Gel chromatography of the gastric tissue extract showed a single peak of FSH immunoreactivity that coeluted with the 125I-labeled highly purified FSH preparation (NIDDK, NIH). Furthermore, the FSH in the pituitary tissue extract had a chromatographic profile similar to that of IR-FSH from gastric tissue, and 125I-FSH labeled highly purified FSH, indicating a close resemblance in their molecular sizes. These results demonstrate that IR-FSH is present in the normal human gastric mucosa. The role of this regulatory petpide in gastric tissue, if any, needs to be investigated.

Testicular dysfunction after cyclophosphamide-vincristine-procarbazine-prednisolone chemotherapy for advanced Hodgkin's disease. A long-term follow-up study.

Gonadal functions were evaluated in 92 male patients after treatment for advanced Hodgkin's disease. The patients received six to ten cycles of cyclophosphamide, vincristine, procarbazine, and prednisolone (COPP) chemotherapy. All patients were in remission and were followed for 1 to 17 years (median, 6). Testicular atrophy was noticed in 89 (96.7%) patients. All patients remained azoospermic during the period of follow-up. The testosterone levels did not differ before and after treatment. The follicle stimulating hormone levels rose from pretreatment values (mean +/- standard deviation) of 179.27 +/- 21.99 ng/ml to 578.79 +/- 102.36 ng/ml after the treatment; the rise was significant (P less than 0.001). The luteinizing hormone levels rose from pretreatment values of 106.96 +/- 20.37 ng/ml to 127.37 +/- 32.19 ng/ml after treatment; the rise was significant (P less than 0.05). Testicular biopsy specimens in 19 patients showed germinal aplasia in all cases. It is concluded that six or more cycles of COPP chemotherapy for advanced Hodgkin's disease in men leads to permanent sterility.

Immunocytochemical localisation of follicle stimulating hormone (FSH) in normal, benign and malignant human prostates.

Immunocytochemical localisation of follicle stimulating hormone (FSH) was carried out in normal, benign and malignant human prostates by indirect immunoperoxidase technique. Positive staining was observed in the epithelial cells of all the three categories, while the stromal cells showed a weakly positive reaction in a few specimens. The brown reaction product was dispersed in the cytoplasm of the epithelial cells. These observations demonstrate the presence of immunoreactive FSH-like peptide in human prostate. The significance of FSH in the aetiopathology of prostatic disorders is discussed.

Immunoreactive-FSH in human normal gastric mucosa.

Using indirect immuno-peroxidase staining technique, localization of immunoreactive follicle-stimulating hormone (IR-FSH) is demonstrated in the cytoplasm of the epithelial cells of normal human stomach. In view of their triangular shape and central nucleus and their predominance in the intermediate glands of the gastric mucosa, these cells are identified as parietal cells. The stromal tissue is devoid of staining reaction.

Conversion to estriol in "normal", benign and malignant human breast tissues.

Since there are controversial reports in the literature on 16 alpha-hydroxylase activity in human breast tumors, we have studied this enzyme activity in "normal", benign and malignant breast tissues by measuring the conversion of estradiol to estriol. Whereas 8 out of 14 (58%) malignant tumors showed positive activity, none of the other tissues had any detectable 16 alpha-hydroxylase activity. These preliminary results, when combined with reports in the literature on animal and human tissues, appear to indicate that more extensive studies on this enzyme could be useful as it could be an additional metabolic marker in human breast cancer.

Prostatic inhibin-like peptide quantified in urine of prostatic cancer patients by enzyme-linked immunosorbent assay.

This is a highly specific enzyme-linked immunosorbent assay (ELISA) for measuring prostatic inhibin-like peptide (PIP) in urine, in which we use penicillinase (EC 3.5.2.6) conjugated with PIP and, as solid phase, a polystyrene microtiter plate. We used this ELISA to measure PIP in 24-h urine specimens from men with prostatic cancer (PCa) and from age-matched controls. For prostatic cancer patients the mean +/- SEM urinary PIP of 36.1 +/- 5 micrograms/24 h was significantly (P less than 0.001) lower than the mean of 127.1 +/- 9 micrograms/24 h for the age-matched controls. PIP values for 30 samples measured by both ELISA and RIA correlated well (r = 0.985). We could detect as little as 1.56 ng of PIP in a sample. Analytical recovery of added PIP ranged from 91% to 104%. Mean CVs were 8.9% within-assay and 12.7% between-assay. We believe that this ELISA will be useful in assessing the status of PIP in men with normal and diseased prostates and in examining the function of the hypothalamus-pituitary-prostate axis.

Serum and urinary prostatic inhibin-like peptide in benign prostatic hyperplasia and carcinoma of prostate.

The levels of immunoreactive prostatic inhibin-like peptide (PIP), having follicle-stimulating hormone suppressing properties, were estimated in the sera and urine samples of patients with benign prostatic hyperplasia (BPH) and prostatic carcinoma (PC) as compared to age-matched controls. Significantly elevated serum PIP levels in BPH (107.8 +/- 19 ng/ml) and PC (88.7 +/- 9 ng/ml) patients were observed as compared to those in control men (10.2 +/- 1 ng/ml). Unlike serum, in urine high levels of PIP in BPH (294 +/- 49 micrograms/24 h) and extremely low levels in PC (23.6 +/- 5 micrograms/24 h) patients were seen as compared to control values (137.6 +/- 10 micrograms/24 h). Furthermore, striking differences were observed between the urinary PIP levels of BPH and PC patients. The results of the present investigation thus indicate the possible use of urinary PIP as a biological marker for prostate cancer.

Modulation of rat prostatic inhibinlike peptide (PIP) by steroids and protein hormones.

Prostatic inhibinlike peptide (PIP) was detected in the ventral but not in the dorsal lobe of rat prostate. On orchiectomy, PIP concentration in the ventral prostate increased significantly, whereas it decreased on testosterone administration and attained value comparable with that in intact control. Estradiol-17 beta treated animals exhibited high levels of PIP in absence of significant alterations in the protein content. The effect of prolactin and human chorionic gonadotropin on PIP concentration was not so prominent at the dose levels studied. The present study thus demonstrates negative regulation of PIP by testosterone and stimulatory effect of estradiol-17 beta on PIP in rat ventral prostrate.

Inhibin-like material--an immunohistologic marker for prostatic origin of metastases.

Immunocytochemical localization of inhibin-like material (ILM) using a specific antiserum generated against ILM of prostatic origin was carried out in metastatic lymph nodes of known primary prostatic tumours and in rectal biopsies showing direct invasion with known prostatic carcinoma. Nine of the 11 metastatic lymph nodes gave positive reaction, which was readily apparent in differentiated tumours showing micro-acinar formation. In poorly differentiated tumours it was often focal and indicated intracytoplasmic staining within randomly scattered cells. Rectal biopsies showing invasion of prostatic carcinoma (4 cases) also showed positive reaction for ILM. Using this experimental protocol, the reaction for metastatic lesions from patients with non-prostatic carcinomas was completely negative indicating the specificity of the test for prostatic origin of metastasis. In conclusion, the present investigation indicates the potential application of ILM in confirming or excluding the prostatic origin of invasive tumour in metastatic lesions.