Parathyroid hormone-related peptide in the human fetal uro-genital tract.

Using a polyclonal antiserum raised against the first 34 amino acids of human parathyroid hormone-related peptide (PTHrP), we have localized PTHrP throughout the uro-genital tract of the human fetus aged between 8 and 40 weeks. Staining was present in the developing mesonephros, metanephros, gonads and in both the adrenal cortex and medulla. In particular, the developing mesonephric and metanephric renal tubules were intensely positive. Using Northern hybridization analysis we have detected a complex pattern of PTHrP mRNA transcripts ranging in size from 1.4 to 4.5 kb in early second trimester human fetal kidney. The presence of PTHrP in the mesonephros and metanephros provides evidence for a role for PTHrP in the regulation of fetal calcium metabolism. However, its presence in the gonad and adrenal gland invites the possibility of a wider role for PTHrP.

GABA-receptors in peripheral tissues.

Gamma-aminobutyric acid (GABA) and its receptors are found in a wide range of peripheral tissues, including parts of the peripheral nervous system, endocrines, and non-neural tissues such as smooth muscle and the female reproductive system. In all these, both GABAA- and GABAB-receptor types are found, with good evidence for a physiological role in the gut, pancreatic islets and the urinary bladder. In some tissues, the pharmacology of GABA-induced actions is quite atypical and should be further explored with the newer ligands and modulators for GABAA- and GABAB-receptors.

[Four putative neuropeptides concentrations in the human urogenital tract. Comparison of the neuropeptides concentration between malignant and benign tissues].

In order to evaluate a possible role of several peptides in the human urogenital tract, peptide concentrations in urogenital tissues collected from surgery were measured using specific radioimmunoassay. The specimens were extracted in boiling 0.5M acetic acid, and these extracts were utilized to measure neuropeptide concentrations, i.e., neuropeptide Y(NPY), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and peptide 7B2. The highest concentrations of NPY were found in seminal vesicle (145 +/- 42pmol/g) and vas deference (104 +/- 26pmol/g). There was no significant difference in NPY concentration between malignant and non-malignant tissues (prostate and urinary bladder). High concentrations of VIP were also observed in several urogenital tissues (seminal vesicle, vas deference and urethra). VIP concentrations in prostatic cancer and carcinoma of urinary bladder seemed to be reduced, though no significant difference could be found in each corresponding tissue. Pituitary peptide 7B2 was found to be present in the human urogenital tract in relatively low concentrations. A significant difference was observed in CGRP concentration between carcinoma of urinary bladder and adjacent normal vesicular tissues (p less than 0.05). These four peptide immunoreactivities were further characterized by gel permeation or high performance liquid chromatography. Each main immunoreactivity in urogenital extracts seemed to correspond to each synthetic standard or pituitary extracts (in case of 7B2). These results demonstrated that pituitary peptide 7B2 was shown to be present in the human urogenital tract and that the distribution patterns of these peptides might correlate to their pathophysiological role in the urogenital tract. Furthermore, the absence of CGRP immunoreactivity in carcinoma of urinary bladder may be useful for additional diagnostic information.

Localization of lactoferrin in the male reproductive tract.

The immunohistochemical localization of lactoferrin in the normal human prostate, seminal vesicle, vas deferens, epididymis and testis was studied using the peroxidase-antiperoxidase method at the light and electron microscopical level. Lactoferrin immunoreactivity was localized in the glandular epithelial cells and granulocytes in the prostate and seminal vesicle. In the prostate, lactoferrin showed an uneven distribution; some of the glands contained exclusively positive cells and others were completely lactoferrin negative, while the rest contained scattered positive cells. The seminal vesicles were divided into three segments, and their lactoferrin content varied significantly although it was always epithelial. The ductus deferens, epididymis and testis contained no lactoferrin. In conclusion, lactoferrin was found in the prostate and seminal vesicles, but not in the testis.

Use of exfoliated cells to study tissue-specific levels of beta-carotene in humans.

This study was designed to explore the feasibility of using exfoliated cells to study beta-carotene incorporation into different epithelial tissues in humans. Exfoliated cells were collected from the oral cavities (by brushing the oral mucosa) and from the urogenital tracts (by centrifuging urine samples) of 36 females and basal levels of beta-carotene (without oral supplementation) were determined. Beta-carotene levels in cells from the two sites differed significantly, although a weak correlation was observed. As a second aspect of the study, 10 of these females were given oral supplementation with beta-carotene (90 mg twice weekly for 4 weeks). Beta-carotene levels increased significantly in both exfoliated urogenital tract (6.8-fold) and oral mucosa (5-fold) cells. However, the supplemented levels remained significantly different for the two types of cells. Beta-carotene levels did not change in individuals receiving a placebo treatment (n = 7). These studies suggest that exfoliated cells collected from different sites may be of value in quantifying tissue levels of beta-carotene during cancer intervention trials.

Detection of Chlamydia trachomatis in culture and urogenital smears by in situ DNA hybridization using a biotinylated DNA probe.

The detection of Chlamydia trachomatis by in situ DNA hybridization in urogenital smears was investigated using a commercially available biotinylated DNA probe. Intracellular staining of inclusion bodies was used as the criterion for positivity. Of 35 patients with a culture proven chlamydial infection 19 had smears in which C. trachomatis was detected by in situ DNA hybridization, indicating a sensitivity of 54%. Of 57 patients with a negative culture, two had positive smears by in situ DNA hybridization. To compare whether the criterion for positivity had influenced the sensitivity obtained with in situ DNA hybridization, 14 duplicate smears from culture positive patients were analysed with in situ DNA hybridization and immunofluorescence. Both methods detected intracellular inclusion bodies in seven of these smears, suggesting that the presence of infected cells mainly determines the sensitivity. The DNA probe did not cross-react with micro-organisms commonly found in the urogenital tract.

Tamm-Horsfall protein outside the kidney.

Tamm-Horsfall protein is the main protein in normal urine and readily precipitates. Previously there has been little study of the protein outside the kidney. An antiserum to the protein was used in an immunohistological investigation of human tissues. No Tamm-Horsfall protein was detected in normal organs other than the kidney. The protein was detected outside the kidney in abnormal organs in three general sites: (i) in ulcers of epithelial surfaces exposed to urine, (ii) deep in the wall of a bladder that had had previous surgery and (iii) in lymph nodes at the hilum of kidneys that had been obstructed or contained extra-tubular deposits of the protein. There was little evidence of a cellular response to the deposits of Tamm-Horsfall protein. The antiserum to Tamm-Horsfall protein is a good immunohistological marker of extravasated urine and is useful in the study of such conditions as fistulas of the urinary tract, operations on the urinary tract and reflux of urine into lymphatics and lymph nodes.

Neuromedin U--a study of its distribution in the rat.

The distribution of neuromedin U, a novel peptide originally isolated from porcine spinal cord, was investigated in the rat using a recently developed radioimmunoassay. High concentrations of neuromedin U-like immunoreactivity were found in the pituitary gland and gastrointestinal tract. Significant concentrations of immunoreactivity were also found in several regions of the rat brain, spinal cord and both male and female genitourinary tracts. In the small intestine, neuromedin U-like immunoreactivity was restricted to the submucosal muscular layers, suggesting localization in neurones rather than in epithelial cells. Chromatographic analysis of pituitary, spinal cord and gut revealed a single peak of immunoreactivity which did not co-elute with either synthetic porcine neuromedin U-25 nor neuromedin U-8, indicating inter-species molecular heterogeneity.

Characterization of a novel prepro VIP derived peptide.

A newly identified, large molecular weight form of peptide histidine methionine (PHM), has been found not only where it was first revealed, in the stomach, but also in high concentrations in the nasal mucosa and urogenital system, though not in the central nervous system, intestine and lung. An antibody to the spacer peptide sequence prepro-VIP 111-122, lying between PHM and VIP, also reacts directly with the large molecular form of PHM. It is suggested that the post-translational processing of prepro-VIP differs between tissues and in some, cleavage may not occur at the C-terminal end of PHM. The biological significance of this is currently unclear.

Immunohistochemical localization of pancreatic secretory trypsin inhibitor in fetal and adult pancreatic and extrapancreatic tissues.

Pancreatic secretory trypsin inhibitor (PSTI) has been thought to be only a secretory trypsin inhibitor of human pancreas, but the serum content of immunoreactive PSTI is elevated without pancreatic disease. Using the peroxidase-antiperoxidase method, immunoreactive cells for PSTI were found in human pancreas, stomach, duodenum, appendix, colon and urinary tract of both fetus and adult, adult gall bladder, and fetal lung. PSTI-immunoreactive cells were identified in fetal pancreas at the tenth gestational week, and in extrapancreatic tissues at the sixteenth (gastrointestinal and urinary tract) and twentieth weeks (lung). PSTI-immunoreactive cells of fetal lung were present in neuroepithelial bodies. Strongly positive cells in fetal duodenum were argyrophilic and resembled endocrine cells. Immunohistochemical study was also performed on tissues associated with inflammatory diseases of gastrointestinal tract. The distribution pattern of immunoreactive cells in the stomach varied in accordance with chronic gastritis. Immunoreactive cells were also found in endocrine micro-nests and in a carcinoid tumor associated with fundic gastritis. These results suggest that PSTI may play some physiological role other than secretory trypsin inhibition of the pancreas.

[Histamine in the urogenital tract]. / Histamin im Urogenitaltrakt.

Tissue histamine determination in the urogenital tract showed that there are significant differences concerning tissue histamine content between intraoperative and postmortem tissue samples. The difference appeared to be most significant in the case of renal pelvis samples, where the decrease was about 90% in the autopsie material compared to intraoperative tissues. In comparing tissue histamine content in special organs of different animal species we discovered considerable differences; e.g. in the ureter of the dog we measured 30 micrograms/g tissue - where as against the values in man amounted to only 6 micrograms/g tissue. The physiological meaning of the different tissue histamine levels is unknown.

Regulatory peptides--the distribution of two newly discovered peptides: PHI and NPY.

PHI (peptide with histidine and isoleucine) and NPY (neuropeptide with tyrosine) are recently discovered regulatory peptides. There are close sequence similarities between PHI and both growth hormone releasing factor (GRF) and vasoactive intestinal polypeptide (VIP) and between NPY and pancreatic polypeptide. Using immunocytochemistry and radioimmunoassay we have revealed the broad distributions of PHI and NPY in neurons of the central nervous system and the majority of peripheral tissues. Tissues which are particularly well provided with these peptides include gut, pancreas, respiratory tract, skin and the genitourinary and cardiovascular systems. In most peripheral tissues, PHI-containing ganglion cells occur locally. NPY-containing fibres originate in part from cell bodies outside the tissues, in the sympathetic nervous system. Comparative studies indicate that PHI and VIP are co-stored in the same neurone and are identically distributed, thus suggesting the existence of a common precursor and subsequent gene duplication. The possible co-existence of catecholamines and NPY, suggested by the consistent finding of very similar distributions of the two substances, was investigated using antibodies to converting enzymes involved in catecholamine synthesis (tyrosine-hydroxylase and dopamine-beta-hydroxylase). The two enzymes and NPY were found together in at least part of the same neuronal system.

Extragastric H2-receptors.

The discovery of H2-receptors by Black and coworkers opened a new era in the history of histaminology. Their importance became apparent when, soon after their discovery, the physiological role of histamine in the regulation of gastric secretion was clarified. In the last decade the explosion of research in the field of histamine and its receptors has demonstrated that H2-receptors have a much wider distribution than previously suspected. H2-receptors are found in the brain, the endocrine and exocrine glands, the pulmonary system, the cardiovascular system of different species, the gastrointestinal muscle, the genitourinary system, the immunological system and in the skin. In some instances stimulation of the various H2-receptors evokes responses that are opposite to those elicited by stimulation of H1-receptors. In other cases they are quite similar. Usually, activation of H2-receptors leads to an increased activity of the adenylate cyclase system with a consequent increase in intracellular cyclic AMP. Most H2-receptors are located postsynaptically on muscle or gland surfaces. However, there is recent evidence concerning the possibility of a presynaptic localization with a modulatory function on the release of different mediators. Finally, controversy exists over the possibility that H2-receptors do not represent a homogeneous population. In fact, several observations suggest that "anomalous" H2-receptors are characterized by different sensitivity to the H2-antagonists and/or antagonists in various tissues. If this is true, exact characterization of H2-receptors will be decidedly more difficult and will require "super selective" H2-agonists and antagonists which are not currently available.

Vasoactive intestinal polypeptide (VIP) in the male genitourinary tract: concentration and motor effect.

Vasoactive intestinal polypeptide (VIP) has previously been shown in nerves of the male and female genitourinary tract, appearing to innervate vascular and nonvascular smooth and epithelial cells. In the present study the concentration of VIP in tissue extracts of different parts of the male genitourinary tract from cat and man was determined by radioimmunoassay. In addition, the effect of VIP on the contractility of the smooth muscle from the cat genitourinary tract was investigated in vitro. The tissue concentrations of VIP were generally higher in cat than in man. In both species high concentrations were found in the vas deferens, bladder, urethra and prostate, In concentration from 3 x 10(-8) to 6 x 10(-7) mol x 1-1, VIP inhibited in a concentration-dependent manner the muscle contractions in specimens from all regions examined, i.e., the vas deferens, ureter, corpus of the bladder, and urethra. The data indicate that VIP might play a physiologic role in the local nervous control of the smooth muscle activity in the male genitourinary tract.

Concentration of substance P-like immunoreactivity (SPLI) in tissues of dog, rat and mouse.

The distribution of SPLI in various tissues from dog, rat and mouse was determined by radioimmunoassay. The gastrointestinal, urogenital and tracheobronchial tracts were found to contain SPLI with marked differences in levels in different parts of these tracts. Glandular tissues, such as salivary glands and pancreas, was well as eye, tongue and skin also contain SPLI. Species differences were encountered both in absolute amounts and the distribution of SPLI. The highest concentrations of SPLI were generally found in mouse followed by dog and rat. The present results are in agreement with those of previous studies in which SP was demonstrated by bioassay technique, but due to the greater sensitivity of the radioimmunoassay it was possible to show a much wider distribution of SP. Virtually all organs in which an effect of SP has been demonstrated also contain SPLI.