Showing papers in "Investigative urology in 1979"

Establishment and characterization of a human prostatic carcinoma cell line (PC-3).

Abstract: The establishment, characterization, and tumorigenicity of a new epithelial cell line (PC-3) from a human prostatic adenocarcinoma metastatic to bone is reported The cultured cells show anchorage-independent growth in both monolayers and in soft agar suspension and produce subcutaneous tumors in nude mice Culture of the transplanted tumor yielded a human cell line with characteristics identical to those used initially to produce the tumor PC-3 has a greatly reduced dependence upon serum for growth when compared to normal prostatic epithelial cells and does not respond to androgens, glucocorticoids, or epidermal or fibroblast gowth factors Karyotypic analysis by quinacrine banding revealed the cells to be completely aneuploid with a modal chromosome number in the hypotriploid range At least 10 distinctive marker chromosomes were identified The overall karyotype as well as the marker chromosomes are distinct from those of the HeLa cell Electron microscopic studies revealed many features common to neoplastic cells of epithelial origin including numerous microvilli, junctional complexes, abnormal nuclei and nucleoli, abnormal mitochondria, annulate lamellae, and lipoidal bodies Overall, the functional and morphologic characteristics of PC-3 are those of a poorly-differentiated adenocarcinoma These cells should be useful in investigating the biochemical changes in advanced prostatic cancer cells and in assessing their response to chemotherapeutic agents

Effects of neonatal steroids on male sex tissues.

Abstract: The effects of neonatally administered sex hormones on the growth and hormone responsiveness of the sex accessory tissues of the adult male rat were investigated. If intact neonatal male rats are treated with estradiol diproprionate, the size of the ventral prostate in the adult animal at day 70 of age is less than 10 per cent of the untreated control value. If androgens are administered to intact neonates, the size of the ventral prostate at day 70 is one-third to one-half of the untreated control values. The androgenic response of these abnormally small prostates was tested by treating these adult animals with testosterone propionate daily from day 70 to day 83. Toward ventral prostates of the animals treated with estrogens in the neonatal period show a very limited response to exogenous androgens at adulthood. In contrast, the ventral prostates of the animals receiving testosterone propionate or dihydrotestosterone propionate in the neonatal period were fully responsive to exogenous androgens. In addition, the ability of the adult prostate to respond fully to androgens requires the presence of the testes in the neonatal period. Comparing neonatal castrates with intact neonates, it was shown that the administration of androgen in the neonatal period was only partially effective in restoring the androgenic responsiveness of the adult ventral prostate. This suggests that other testicular factors, or the presence of testosterone at other time periods before the onset of puberty, may be necessary for the normal androgenic induced response of the adult sex accessory tissues.

Prostatic osteoblastic factor.

Abstract: Cell cultures of fetal rat osteoblasts and organ cultures of fetal rat calvaria were used to study the effects of extracts of prostatic tissue on bone cell growth. Extracts of bening prostatic hyperplasia, but not an undifferentiated prostatic carcinoma, stimulated the incorporation of 3H-thymidine and 14C-proline into fetal rat osteoblasts, calvaria, and skin fibroblasts. Extracts of well-differentiated prostatic cancer and normal postpubertal prostate also stimulated 3H-thymidine incorporation by fibroblasts. These findings support the hypothesis that a potentially unique factor found in prostatic tissue induces the osteoblastic response of bone to metastatic prostatic cancer.

Attenuation by monosodium urate of the inhibitory effect of glycosaminoglycans on calcium oxalate nucleation.

Abstract: The interaction between mucopolysaccharide (glycosaminoglycans) and monosodium urate, with respect to their effects on spontaneous precipitation of calcium oxalate, was explored in vitro. Heparin and, to a lesser extent, chondroitin sulfate were found to inhibit calcium oxalate precipitation. The formation product ratio (FPR, limit of metastability or the minimum supersaturation required for spontaneous nucleation) of calcium oxalate was increased 29 per cent by 0.05 mg of heparin per liter, and by 70 per cent with 1 mg of heparin per liter. The increment produced by heparin was much less pronounced when the solution containing the mucopolysaccharide was preincubated with crystalline monosodium urate. Pretreatment of the solution containing heparin (0.05 mg per liter) by monosodium urate (0.5 mg per ml) virtually eliminated the inhibitory action of heparin. Studies with radioactive heparin indicated that monosodium urate binds heparin. In contrast, the inhibitory effect of chondroitin sulfate was unaltered by prior exposure to monosodium urate. The results indicated that monosodium urate may attenuate the inhibitor activity of certain mucopolysaccharides such as heparin and facilitate the calcium oxalate precipitation. The findings provide further support for the important etiologic role of hyperuricosuria in the formation of calcium oxalate stones.

Sacral root stimulation in controlled micturition. Peripheral somatic neurotomy and stimulated voiding.

Abstract: Stimulation of the sacral ventral roots with the largest detrusor representation was found to be moderately successful for bladder emptying in acute canine experiments. However, removal of spinal reflexes arising from sacral root stimulation did not eliminate sphincteric resistance. For that reason ventral roots were stimulated after peripheral somatic neurotomy. Unilateral pudendal nerve sectioning was compared with selective sectioning of the somatic fibers of S2, the root with the greatest detrusor representation in canines. Both were equally effective in significantly reducing outlet resistance. Selective nerve section has one advantage: it preserves more of the nerve pathways involved in reflexogenic erection.

Urinary bladder and sphincter responses to stimulation of dorsal and ventral sacral roots.

Abstract: The sacral spinal roots have a long intraspinal yet extradural pathway that makes them ideal for electrode implantation for the control of micturition. In the hope of identifying a specific motor pathway to the bladder within these roots, a study of 10 mongrel female dogs was undertaken: bladder and sphincter responses to stimulation of individual sacral roots, and to stimulation of the dorsal and ventral components of each root, were recorded urodynamically. The root producing greatest detrusor response was then divided into its motor and sensory components, and the central and peripheral ends of the latter were stimulated. We found that sphincter resistance to bladder emptying was diminished by removing the afferent dorsal roots from the stimulus field. The physiologic implications of this finding are discussed.

Histologic study of the early events in oxalate induced intranephronic calculosis

Abstract: The early phases of calcium oxalate crystal formation in rat kidneys after intraperitoneal challenge with sodium oxalate were examined. From the results we conclude that the crystals are formed intraluminally in the proximal tubules of the renal cortex. The size, number, and distribution of calcium oxalate particles depend on the amount of sodium oxalate injected and the time interval after its injection. Tubular necrosis and dilation were evident after administration of higher doses of sodium oxalate. The papillary tip and corticomedullary junction were the preferential sites for crystal localization after longer periods of injections of higher doses of sodium oxalate.

Crystallization studies in a urothelial-lined living test tube (the catheterized female rat bladder). I. Calcium oxalate crystal adhesion to the chemically injured rat bladder.

Abstract: We developed an in vivo technique to study crystallization in a urothelial-lined "living test tube" by employing the catheterized female rat bladder. Calcium oxalate crystals adherent to the bladder urothelium were distinguished from free or intraluminal crystals. Chemical injuries of the urothelium induced by either 0.1 N HCL or 5 per cent Triton X 100 in saline resulted in marked calcium oxalate crystal adhesion to the injured urothelium. Control bladders without urothelial injury remained free of adherent crystals but formed large numbers of small free crystals relatively uniform in size. Histologic and fresh microscopic evaluations of urothelial injury and crystal adhesion are presented.

Attachment of bacteria to exfoliated cells from the urogenital tract.

Abstract: To establish urogenital infections, organisms must adhere to the mucosal lining. A differential adherence capacity among various bacterial species was observed when exfoliated urethral and urothelial cells were tested in an in vitro system. No difference in the adherence capacity of a particular species was observed when tested with exfoliated cells obtained from voided urine from different healthy individuals of the same sex. Escherichia coli harvested directly from urine specimens of patients with significant bacteriuria showed a significantly higher capacity to adhere than when obtained from the primary isolation plate. Staphylococcus saprophyticus adhered significantly better to urothelial cells than did Staphylococcus epidermidis. Adherence did not differ when the tests were performed in ultrafiltrated, infected and noninfected urine. Variations of the osmolality did not influence the adherence rate of E. coli. Gonococci showed an increased capacity to adhere when tested in urine of increasing acidity. Gonococci producing T1 colonies adhered by significantly higher numbers per cell than such bacteria producing T4 colonies.

Rapid screening for urease inhibitors.

Abstract: Two methods are described that are suitable for the rapid screening of compounds as urease inhibitors. The first utilizes an electrode sensitive to NH4+ ions; the second is dependent on pH rise. A feature of both is a direct readout of reaction rate.

Observations on the gubernaculum during descent of the testis.

Abstract: The mechanisms that influence the descent of the testis are not clearly understood. The gubernaculum is a structure worthy of scrutiny inasmuch as it is conspicuous during descent, but virtually disappears after descent is complete. Early in gestation, the rat gubernacular bulb consists of loose mesenchymal cells that develop into fibrillar cells. These later thicken into rhabdomyoblasts that, near the end of gestation, differentiate into spiral striated muscle bundles, and eventually migrate outward into the abdominal/scrotal wall. The rhabdomyoblasts of the female gubernaculum do not differentiate further but rather undergo fatty degeneration. It is possible that spiral contractions of the attached gubernaculum produce tension on the testis and induce descent. The gubernaculum as the receptor organ for testicular descent may be responsive to local testicular hormones. Likely candidates are testosterone, dihydrotestosterone, or Mullerian Inhibiting Substance. A thorough knowledge of the sequential differentiation of the gubernaculum during embryonic development sets the stage for the study of its response to hormonal manipulation both in vivo and in vitro.

An electron microscopic study of the interaction between vesical epitherlium and E. Coli.

Abstract: Interaction between rat bladder epithelium and Escherichia coli was observed using the transmission electron microscope. As a first step, the bacterial cell coat attaches to the cell coat of the luminal membrane. Then, the luminal membrane enfolds to envelope the bacterium and take it within the cytoplasm. The unit membranes covering the bacteria are destroyed and some of these bacteria are found to be deformed and probably dead in the cytoplasm. Finally, the superficial epithelial cells invaded by many bacteria are collapsed and desquamated into urine. However, these epithelia show a rather prompt recovery and form the normal three layers within 24 hr after inoculation. This experiment suggests one of the defense mechanisms of the vesical epithelium againts E. coli. It is likely that the onset of clinical cystitis may be prevented by the same mechanism during the occasional bacterial invasion of the bladder.

Ureteral plication. A new concept in ureteral tailoring for megaureter.

Abstract: Currently, the most favored surgical procedure used to restore normal peristalsis to massively dilated adynamic ureters is ureteral tailoring This procedure, however, seriously jeopardizes the ureter's blood supply and frequently results in ischemia with ureteral stenosis and further renal deterioration To preserve the distal ureter's vasculature a new technique of reducing ureteral caliber and restoring normal peristalsis with minimal disruption of the blood supply was developed and performed in 14 dogs with excellent results

Citrate in daily and fasting urine: results of controls, patients with recurrent idiopathic calcium urolithiasis, and primary hyperparathyroidism.

Abstract: In three groups--patients with recurrent calcium urolithiasis (RCU), patients with primary hyperparathyroidism (pHPT), and healthy controls--citrate was measured enzymatically in 24 and in 2-hr urine after an overnight fast. Citrate excretion per 24 hr was significantly lower in RCU than in age and sex matched controls, whereas there was no significant difference in citrate excretion in urines from the 2-hr morning collection. In pHPT citrate was also lower than in controls and fell within the range of RCU of comparable age. Both categories of urines (24 and 2hr) have in common the characteristic that the actual citrate concentration is lower by 50 per cent in RCU and pHPT than in controls, mainly as a result of the higher urine volume. Correction of citrate for creatinine does not disclose further differences among the populations studied but conversely hampers exact interpretation of urinary citrate in the absence of strict separation of individuals according to sex and age. From these data we conclude that (i) a low excretion and concentration of urinary citrate is detectable in calcium lithiasis and may contribute to a deficiency in inhibitory activity against nucleating processes in stone-forming urine; and (ii) the differences in urinary citrate elicited in samples of 24 and 2-hr morning urine are of unknown origin and merit further investigations.

Quantitative analysis of alpha and beta adrenergic receptor densities in the lower urinary tract of the dog and the rabbit.

Abstract: Using specific radioligand receptor binding assays, we have, for the first time, quantitated both alpha- and beta-adrenergic receptors in the musculature of the lower ureter, bladder body, posterior bladder base, and proximal urethra of the dog, and in the bladder base and body of the rabbit. For the beta receptor assay 3H-dihydroalprenolol was used as the ligand, and, for the alpha receptor assay, 3H-dihydroergocryptine was used. In the dog, there was a significantly greater density of beta receptors in the bladder body than in either the ureter or the posterior bladder base. The urethra had an intermediate density. The bladder base and the proximal urethra had a significantly greater density of alpha receptors than either the ureter or the bladder dome. The dissociation constant of beta receptors for DHA was approximately 9 nM for all tissues studied, and that of alpha receptors for DHE approximately 7 nM. A similar distribution of alpha-and beta adrenergic receptors was seen in the rabbit bladder.

Human prostatic acid phosphatases: purification of a minor enzyme and comparisons of the enzymes.

Abstract: The minor enzyme of human prostatic acid phosphatases (pI 5.5) with high specific activity (orthophosphoric monoester phosphohydrolase, acid optimum, EC 3.1.3.2) has been purified for the first time as a pure enzyme protein. The enzyme was a single protein when examined by polyacrylamide gel electrophoresis and isotachophoresis. The specific activity was 1080 micromole per (min X mg) for hydrolysis of 5.5 mmole per liter of p-nitrophenylphosphate at pH 4.8 and 37 C. The purification coefficient was 540 and the recovery of enzyme activity was 2 per cent. The molecular weight of the enzyme subunit when measured by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate was 54,000. The Km of the purified enzyme was 3 X 10(-4) mole per liter for p-nitrophenylphosphate. An antiserum to this enzyme was prepared. The enzyme was cross-reactive with the main enzyme (pI 4.9) of human prostatic acid phosphatases in immunoelectrophoresis. No precipitin arc with the acid phosphatase in the serum of a prostatic carcinoma patient could be shown. Antiserum to the main enzyme caused a precipitin line with the same serum sample.

The synthesis and release of prostaglandins by human urinary bladder muscle in vitro.

Abstract: Using the technique of bioassay the presence, production, and importance of prostaglandins (PG) in human bladder muscle have been investigated in vitro. Low basal tissue levels of PG were shown. E series PGs were released by isolated bladder muscle strips in organ bath experiments. The spontaneous activity and PG release of these strips could be markedly reduced by the addition of indomethacin. Incubation of bladder muscle with the PG precursor, arachidonic acid, resulted in the synthesis of substantial quantities of PGs.

On the possible role of glycosaminoglycans as natural inhibitors of calcium oxalate stones.

Abstract: An extract of glycosaminoglycans was prepared from normal urine and components fractionated by electrophoresis on Vinylite support medium. Several fractions displayed an ability to inhibit formation of calcium oxalate crystals in vitro. Inasmuch as well recognized urinary glycosaminoglycans such as chondroitin sulfates and hyaluronic acid must contribute to the urine inhibitory power, an unknown material was isolated with potent inhibitory properties. The material contained little detectable uronic acid of hexosamine and had a molecular weight greater than 10,000.

A study of the phase transformation of calcium oxalate trihydrate-monohydrate.

Abstract: The precipitation of calcium oxalates may involve transformation from a metastable hydrate such as calcium oxalate trihydrate, COT, to the thermodynamically stable monohydrate, COM. A highly reproducible seeded crystal growth method has been used to study this transformation reaction over a range of temperatures and in ionic media such as the chlorides of sodium, potassium, and magnesium. After the initial dissolution of COT, the calcium concentration remains constant for prolonged periods while appreciable nucleation and growth of COM takes place concomitantly with COT dissolution. The transformation is markedly dependent on temperature, composition, and concentration of ionic media. In urines, the hydrate conversion is appreciably slower and the rate depends upon the method of pretreatment. In these cases, evidence is advanced for the formation of other phases such as the calcium phosphates.

Release of prostaglandins into the pelvic venous blood of dogs in response to vesical distension and pelvic nerve stimulation.

Abstract: Prostaglandin release into the pelvic venous blood in response to vesical distension and pelvic nerve stimulation was studied in anesthetized female nonpregnant dogs. The change in the prostaglandin content of the plasma at sequential physiologic points in the voiding cycle and with pelvic nerve stimulation was measured using a radioimmunoassay technique. A significant liberation of PGE and PGF2 alpha was observed during maximal bladder distension and after micturition. The increased levels returned to near control levels when measured 5 min after bladder evacuation. After pelvic nerve stimulation, PGF2 alpha was also significantly released. The source, mechanism, and functional significance of prostaglandin release during bladder activity are discussed.

Effects of estrogen upon the fine structure of epithelium and stroma in the rat ventral prostate gland.

Abstract: We studied the effects of estradiol upon the fine structure of the rat ventral prostate, with special emphasis on the interstitial tissue. Intact and castrated rats were injected with estradiol or testosterone in combination, or with peanut oil (control). Estrogen exerted an antiandrogenic effect on the epithelium of intact animals resulting in lower epithelial height and reduction in the number of cell organelles and secretory bodies; however, the prostate gland did not atrophy to the same degree as that in castrated controls. In castrates, estradiol had no discernible effect upon the glandular epithelium. Although testosterone alone was able to restore glandular morphology to normal in these animals, estrogen in combination prevented regeneration of rough endoplasmic reticulum and elicited the formation of large lipid-like inclusions and the accumulation of secretory bodies in the apical zone of many cells. Intact animals treated with these hormones in combination exhibited many of the same effects except that a more normal configuration of rough endoplasmic reticulum was present. In castrates, the prostatic stroma became thickened, with a large increase in fibrous material between and surrounding each acinus, although smooth muscle cells retained their normal cytology. With estradiol treatment, singly or in combination with testosterone, smooth muscle cells increased in size and number and organelles decreased in number, cytoplasm became more electron dense, and nuclei became more heterochromatic. Surface vesicles were profuse in smooth muscle cells in animals treated with estradiol alone; larger phagocytic-like vacuoles were characteristic of the glands of animals treated with estrogen and testosterone in combination. In these latter treatment groups, testosterone prevented the overall hypertrophy and hyperplasia of the interstitial tissue; to a degree, endogenous testosterone levels in intact animals treated with estradiol alone prevented these effects as well.

The prostatic carcinoma in laboratory animals: a bibliographic survey from 1900 to 1977.

Abstract: This survey was performed in relation to a study of the role of endocrine glands in rodents that develop prostatic cancer. It is our belief that the data synthesized here can save much tedious work and time, and provide accurate information for investigators in the field of experimental and clinical prostatic cancer.

FSH and LH elevation after radiation for treatment of cancer of the prostate.

Abstract: Regional radiation treatment of prostatic carcinoma produced significant increases in serum follicle-stimulating hormones at 3 and 12 months posttreatment from 6.5 +/- 0.5 to 15.9 +/- 0.9 mIU per ml and 19.5 +/- 1.2 mIU per ml (X +/- SE) respectively. Serum luteinizing hormone also increased at 3 months from 4.5 +/- 0.4 to 6.1 +/- 0.5 mIU per ml (X +/- SE). There were no changes in sex steroid-binding globulin, testosterone, or prolactin. The data indicate damage to the testes as a result of scatter from radiation treatment to the prostate.

Tunica vaginalis autografting for Peyronie's disease: an experimental study.

Abstract: Groups of dogs were studied at varied intervals after autografting of tunica vaginalis from the testes to cover excised patches of tunica albuginea of the penis. Postoperative cavernosography revealed neither ballooning nor chordee. Histologically, the grafts were well accepted and uniformly merged with the neighboring collagen tissues at 6 months. We propose that tunica vaginalis would be ideally suitable for autografting to the penis after excision of Peyronie's plaques.

Micturition and the male genitourinary response to sacral root stimulation.

Abstract: The sacral roots, because of their anatomic and functional organization, are well suited to chronic stimulation. Data from our previous experiments on female dogs suggested that effective voiding responses to stimulation could be obtained by using the ventral root of S2, the sacral nerve in canines with the greatest detrusor representation. Elimination of dorsal root afferents from the stimulus field was essential to the minimization of spinal reflex activity. A selective somatic neurotomy was also necessary to attenuate maximally sphincter activity. In the male, the presence of the prostate gland, the greater urethral length, and the erectile response to stimulation might result in compression of the urethra, compromising micturition. Sacral root stimulation was thus studied using both spinalized and nonspinalized male dogs; the results are discussed with a view toward the applicability of sacral root stimulation to effect micturition in male paraplegics, who comprise the largest percentage of this group of patients.

Effects of clonidine (Catapresan) on urethral pressure.

Abstract: Effects of clonidine (Catapresan) on urethral closure pressure profile were investigated in five patients with minor urologic complaints. Plasma norepinephrine was measured as an index of sympathetic nervous activity. Intravenous injection of clonidine resulted in a pronounced fall in maximum urethral pressure. Plasma norepinephrine concentration decreased as expected. There was a statistically significant correlation between changes in plasma norepinephrine and urethral pressure after clonidine. These findings are discussed in relation to the known effects of alpha-adrenoceptor blocking drugs on urethral pressure. Clonidine may be useful in the treatment of some patients with functional infravesical obstruction.

Pharmacokinetic studies of oxalate in man.

Abstract: The observation of the plasma concentrations after rapid intravenous administration of inulin and 14C-oxalate to six normal and one hyperoxaluric subjects allowed estimations of the half-life of elimination of oxalate (mean 91.7 min), volume of distribution of oxalate (mean = 32.5 liters), calculation of the miscible oxalate pool (mean 3.7 mg), of the plasma oxalate concentration (mean = 11.1 microgram per 100 ml), of the oxalate clearance (252 ml per min), and the determination of the oxalate turnover rate (0.027 mg per min). The 14C-oxalate/inulin clearance ratio was 2.44 implying an additional excretion mechanism apart from glomerular filtration. Inasmuch as 97.2 per cent of the administered tracer were recovered unchanged in urine, nonrenal loss of oxalate is not an appreciable factor. The data of one hyperoxaluric patient are compared with those of the healthy subjects.