Akhouri A. Sinha

Immunoelectron microscopic localization of prostatic-specific antigen in human prostate by the protein A-gold complex.

Human prostate specimens from 25 patients provided 15 normal, four benign prostatic hypertrophy (BPH), and seven adenocarcinoma samples which were studied using a polyclonal antibody against prostatic specific antigen (PSA) and protein A‐gold complex. Our study showed the presence of gold particles in cytoplasmic vesicles and granules, rough endoplasmic reticulum, and occasional lysosomal dense bodies in columnar or cuboidal cells of acini from normal, BPH and well‐differentiated cancerous specimens, but not in the acinar basal cells. Some moderately differentiated and most poorly differentiated tumors contained undifferentiated neoplastic cells in which PSA localizations often were associated with membranous structures since specific cytoplasmic organelles were not differentiated. In prostatic stroma, invasive cells which were of differentiated type localized gold particles. In addition, some neutrophils and macrophages also localized PSA suggesting their role in phagocytosis of extra‐cellularly released PSA in the stroma of BPH and cancerous samples. Thus far, the authors have not observed neutrophils and macrophages with gold particles in normal prostate. We suggest that neutrophils and macrophages were involved in transport of some extracellularly released PSA from the prostatic stroma or other metastatic sites to sera in BPH and cancer patients.

A critical analysis of tumor morphology and hormone treatments in the untreated and estrogen‐treated responsive and refractory human prostatic carcinoma

This study was done to compare the ultrastructural features of prostatic cancer cells in both untreated and estrogen‐treated responsive and refractory patients. Analysis of previously untreated and estrogen‐treated carcinomas showed that the tumors possessed well‐ and poorly‐differentiated acini, and invasive cells. Malignant acini contained numerous columnar (secretory) cells in untreated, but few in treated individuals. Two distinct types of basal cells were observed in untreated and treated acini: type I (light) and type II (dark) cells. In both untreated and treated tumors, type I cells were characterized by having round nuclei with many small aggregates of euchromatin, large nucleoli, and electron‐lucent nucleoplasm. The type II cells had highly pleomorphic nuclei, folded nuclear envelope—sometimes deficient in localized areas, euchromatin, many small aggregates of heterochromatin, large pleomorphic nucleoli, and relatively electron‐opaque nucleoplasm. In some sections, both types of basal cells penetrated through the acinar basal lamina and became invasive. Prostatic carcinomas which were or subsequently became refractory to estrogens showed more abundant type II basal cells than responsive patients. It is postulated that the type II basal cells as well as some type I basal cells are endocrine unresponsive from the outset. Furthermore the tumor possesses a heterogeneous population of cancer cells. While androgen‐dependent tumor cells such as columnar cells may be destroyed by endocrine therapy, these endocrine unresponsive cells continue to proliferate, metastasize, and kill the patient. Therefore, we suggest that patient with advanced prostatic carcinoma initially may be given endocrine therapy to reduce tumor burden caused by endocrine‐sensitive cells. In addition, early treatment with chemotherapy or radiation may be used to destroy unresponsive endocrine‐insensitive cells, before these cells lines have a chance to proliferate and to develop into refractory carcinoma. Cancer 40:2836‐2850, 1977.

Prediction of Pelvic Lymph Node Metastasis by the Ratio of Cathepsin B to Stefin A in Patients with Prostate Carcinoma

Pathologic grade and/or histologic score, extraprostatic extension indicated by invasion of the prostatic capsule, margin, and/or seminal vesicles by prostate cancer cells, serum total prostate‐specific antigen (PSA), free PSA, complexed PSA levels and/or their ratios, regional pelvic lymph node metastases, and clinical staging have been used to diagnose and monitor the treatment of prostate carcinoma (PC) patients. The Gleason grading system is also used to grade/score a patients stage of disease, with lower to higher scores indicating progression of PC. However, Gleasons system cannot be used to distinguish biologically aggressive PCs within a single Gleason score. Our objective was to identify subpopulations (or clones) of aggressive prostate cancers within an individual Gleason score by utilizing biological molecule(s) that also facilitate cancer cell invasion to prostatic stroma and metastasis to the lymph nodes.

Human prostate tumor angiogenesis in nude mice: metalloprotease and plasminogen activator activities during tumor growth and neovascularization of subcutaneously injected matrigel impregnated with human prostate tumor cells.

A critical aspect for growth of solid tumors is the development of a blood supply. Our objective was to establish a model for the study of angiogenesis of human prostate tumors by examining the growth of microvessels into Matrigel containing human prostate tumor cells implanted subcutaneously in nude mice.

Ultrastructure of human amnion and amniotic plaques of normal pregnancy

SummaryThe fine structure of amniotic and amniotic-plaque epithelia has been studied from normal term pregnancies. The columnar/cuboidal amniotic epithelial cells usually have apical or central nuclei, some free ribosomes, patches of granular endoplasmic reticulum, juxtanuclear Golgi complexes, rod-shaped mitochondria, lipid droplets and some glycogen granules. They have short, blunt microvilli which frequently branch and bathe in the amniotic fluid. The lateral plasma membranes enclose tortuous intercellular spaces which are always interrupted by variously folded processes and desmosomes. The epithelial cells rest on a basal lamina and exhibit highly folded basal processes. The amniotic epithelial cells are neither distinctly Golgi and fibrillar types nor “light” and “dark” in appearance.Amnion from near the umbilical cord contains many microscopic and several large plaques. Similar structures are not found on the reflected amnion. The microscopic plaques are whitish and translucent, whereas the large ones are opaque. The large plaques vary between 1–3 mm in diameter, and are over 15 cell layers thick. Each large plaque has a main central region and edges continuous with either the microscopic plaque or the simple amniotic epithelium. The main region shows four zones, namely, stratum basale, stratum spinosum, stratum granulosum and stratum corneum. Such zones are not distinct at the edges. The fine structure of basal cells compares with the amniotic epithelial cells, but the cells of spinosum and granulosum layers possess variable amounts of tonofibrils, keratohyalin granules, free ribosomes and other cytoplasmic organelles and inclusions. The corneum cells are keratinized and are frequently separated by intercellular spaces. They slough into the amniotic cavity singly or as a sheet, and contribute towards the composition of the amniotic fluid. The plaques are of amniotic origin, and are not formed by adhesion of either squamous cells or fetal skin cells (masses of keratinized squames). The present observations suggest that the occurrence of amniotic plaques is normal. The presence of plaques may not be necessarily associated with fetal abnormality. However, increase in numbers of plaques may be caused by conditions of fluid imbalance. The homology and significance of plaques in eutherian mammals have been discussed.

Analysis of Fixation Effects on Immunohistochemical Localization of Prostatic Specific Antigen in Human Prostate

We studied human prostatic specific antigen (PSA) localization in human prostate to investigate the possibility that the previously reported variations in the intensity of antigen staining were due to fixation and embedding methods. We have evaluated the effects of physical and several chemical fixatives on prostate samples obtained immediately after prostatectomies and radical cystectomies. Our analysis of fixation effects and immunohistochemical staining of polyclonal antibody to PSA indicates that the formalin fixation and paraffin embedding methods used previously did provide optimum localization of the antigen and the variations in the intensity of PSA staining could not be attributed to the methodology. Although PSA staining was relatively uniform in the lower grade neoplastic tumors, the higher grade, moderately to poorly differentiated tumors showed intense-through-weak PSA localization or no PSA staining suggesting that PSA staining intensity was not uniformly related to tumor differentiation.

The in vitro localization of H3 estradiol in human prostatic carcinoma. An electron microscopic autoradiographic study

The in vitro localization of H3 estradiol in human prostatic carcinoma was studied in five patients who had not been treated with hormones previously. The isotope was incorporated into the basal and invasive cells but rarely in the differentiated ones of carcinomatous acini. In basal and invasive cells, the grains were localized on the nuclear membranes and associated condensed heterochromatin, and on some mitochondria. The binding of estradiol in prostatic tissues appeared covalent and nonreversible in our preparative techniques for autoradiography. It is tentatively suggested that cytoplasmic binding of estradiol in mitochondria may be associated with the nucleic‐acid‐protein complex found with the organelle membranes. The nuclear binding of the hormone appears associated with the protein portion of the heterochromatin material. Furthermore, the in vitro study has shown that estrogen directly affects some of the prostatic cancer cells within two hours, but the extent of changes in the tumor cells remains undetermined.

Effect of vitamin E deficiency on the growth and secretory function of the rat prostatic complex.

Increased intake of vitamin E has been suggested to be protective against prostate cancer in men, but the effects of vitamin E on prostate growth and function remain poorly defined. The purpose of this study was to determine the effects of vitamin E deficiency on pubertal growth and maturation of the prostate in the rat. Animals were placed on a vitamin E deficient diet at 28 days of age and were followed for 15 and 26 weeks. Vitamin E deficient rats had a circulating vitamin E level of less than 1% of control animals and experienced a decrease in body and testis weight. The deficiency did not alter the weights of the ventral and dorsal lobes of the prostate. However, there was an increase in weight, DNA, and protein contents of the lateral lobe in control and vitamin E deficient rats from 15 to 26 weeks of treatment, but these increases were significantly lower in vitamin E deficient 26-week treated rats. The volume of secretion per milligram tissue was greater in the ventral than lateral or dorsal lobes. The volume of secretion and activity of the secretory 26 kDa protease in the ventral prostate was lower in vitamin E deficient rats at 15 weeks, but not at 26 weeks of treatment. In contrast, the relative protein content of lateral lobe secretion increased in both control and vitamin E deficient rats from 15 to 26 weeks of treatment. The lateral, but not ventral or dorsal, lobes of both control and vitamin E deficient rats were affected by chronic prostatitis as evidenced by infiltration of inflammatory cells. The lateral lobes also showed markedly elevated activities of the matrix metalloproteinases gelatinase A (MMP-2) and gelatinase B (MMP-9). These data indicate that vitamin E deficiency does not alter the growth of the prostatic lobes, nor the onset and extent of lateral lobe specific prostatitis, but it may delay some differentiated functions such as secretion of specific proteins in the ventral lobe. Thus, the effects of vitamin E in the prostate of the rat appear to be selective.

Ultrastructural changes in granulosa lutein cells and progesterone levels during preimplantation, implantation, and early placentation in the western spotted skunk

SummaryThe ultrastructure of corpora lutea obtained during the preimplantation, implantation and early postimplantation periods has been studied in 20 western spotted skunks. Fine structure of granulosa lutein cells was correlated with progesterone levels. The corpus luteum of the prolonged (7 month) preimplantation period contained undifferentiated small granulosa cells and differentiated large granulosa lutein cells. The former ranged in size between 12 and 20 μ and the latter between 20 and 45 μ. The ratio of small and large cells was about equal in an animal 2 days prior to nidation whereas only few small cells and numerous large cells were observed in an animal estimated to be 8 to 12 hours from nidation. Occasionally small cells were observed amidst large ones during the 24 hour nidation period, i.e. adhesion of trophoblast with the luminal uterine epithelium, but small cells were absent in animals after this period. Small cells had some smooth and rough endoplasmic reticulum, rod-shaped mitochondria with platelike cristae, small Golgi complex, and relatively smooth plasma membranes. Large lutein cells had abundant smooth endoplasmic reticulum, membranous whorls of smooth endoplasmic reticulum, usually round mitochondria with tubular and lamellar cristae, a well developed Golgi complex, variable amounts of lipid droplets, and highly plicated and ruffled plasma membranes. Peripheral plasma progesterone levels during the prolonged preimplantation period ranged between 1.1 and 7.9 ng/ml, but during implantation it was between 8 and 16.6 ng/ml. It is suggested that plasma progesterone levels fluctuate during the time of implantation and should not be regarded as a basis to predict actual nidation in the western spotted skunk.

Prostate specific origin of dipeptidylpeptidase IV (CD-26) in human seminal plasma.

PURPOSE A number of peptidases which can metabolize certain bioactive peptides and growth factors have been identified in seminal plasma. Our goal in this study was to determine molecular properties and the tissue source(s) for one of these peptidases, dipeptidylpeptidase IV (DPP IV), in human seminal plasma. MATERIALS AND METHODS We measured the activities of DPP IV with the dipeptide glycylprolyl-p-nitroanalide and its molecular forms using immunoblotting of seminal plasmas of men who were vasectomized or with different sperm concentrations, and in prostatic and seminal vesicle secretions of men undergoing prostatic surgery. RESULTS DPP IV in seminal plasma of vasectomized men was a membrane associated dimer comprised of subunits of approximately 110 kDa. Its activity did not differ in seminal plasmas of vasectomized, azoospermic, oligozoospermic and normozoospermic men indicating no correlation with the concentration of sperm originally present in the semen. The DPP IV antigen (CD -26) and enzymic activity were present in prostatic secretion, but absent from that of the seminal vesicles. These data indicate that the prostate gland is the primary source of DPP IV activity in seminal plasma. There was little variation in its activities in repeat seminal plasma samples from the same individual, and there was no change in its activity with age to 50 years. CONCLUSIONS DPP IV in seminal plasma was derived from the prostate gland and it may be useful as a bioindicator of prostate function and/or disease with age in men.