Beta microseminoprotein is not a prostate-specific protein. Its identification in mucous glands and secretions.

Beta microseminoprotein (beta inhibin, PSP94), an unglycosylated protein of 94 amino acids with unknown function, is one of the predominating proteins in the secretion of the human prostate gland. In this work the authors have demonstrated that the expression of beta microseminoprotein is not restricted to the prostate and that the protein has a previously unrecognized widespread occurrence in the human body. According to radioimmunoassay, beta microseminoprotein immunoreactivity is present in many nonprostatic body fluids. The highest concentrations were found in secretions from the respiratory tract; in tracheobronchial fluid sometimes even at concentrations comparable to that in seminal plasma (about 1 g/l). Intermediate concentrations were found in gastric juice and some samples of secretion from the uterine cervix, whereas tears, saliva, pancreatic juice, bile, and mucus from the colon had low concentrations. According to gel chromatography, the molecular size of the beta microseminoprotein immunoreactivity present in tracheal fluid, gastric juice, and secretion from the uterine cervix did not differ from that of beta microseminoprotein in seminal plasma. The beta microseminoprotein immunoreactive component present in gastric juice had the same amino-terminal amino acid sequence as prostatic beta microseminoprotein (14 residues identified in material purified from gastric juice), providing further evidence for chemical identity of a nonprostatic beta microseminoprotein with the prostatic protein. Immunohistochemical staining with affinity-purified antibodies demonstrated the presence of beta microseminoprotein in many tissues, including the goblet cells in the tracheobronchial epithelium, tracheobronchial submucosal glands, certain mucosal cells in the antrum of the stomach, some glands of Brunner in the duodenum, and in parts of the mucosa of the colon. At least in the respiratory tract, the staining was localized to mucus-containing cells. beta microseminoprotein immunoreactivity also was localized to the cilia of the ciliated epithelium in the respiratory tract, the fallopian tubes, and the Gartner ducts of the uterine cervix. The pattern of tissue distribution of beta microseminoprotein found in this work indicates a connection of beta microseminoprotein with mucous secretions.

Distribution of neurotensin-like immunoreactivities in porcine and human gut.

The distribution of neurotensin-like immunoreactivity (NTLI) in porcine and human intestine was studied by extraction of mucosal and muscular layers of esophagus, stomach, duodenum, jejunum, ileum, and colon. NTLI was quantitated and characterized by radioimmunoassays and gel filtration chromatography. Porcine tissue was obtained in anesthetized animals (n = 6) and human tissue during surgery (n = 28). Concentrations of NTLI increased gradually from the distal esophagus to the ileum. Highest concentrations were found in 2.0 M acetic acid extracts of proximal ileal mucosa (150 (131-223) and 525 (500-729) pmol/g wet tissue, respectively (medians and interquartile range]. After acid extraction, concentrations of intact NT and COOH-terminal and NH2-terminal NTLI were similar, but in water concentrations of NH2-terminal NTLI were high and intact NT and COOH-terminal NTLI low. The distribution of NTLI was similar in the two species. Gel chromatography of ileal, jejunal, and duodenal mucosa indicated that in these tissues NTLI consisted primarily of intact NT. In antral mucosa COOH-terminal immunoreactivity different from NT was detected. The chemical identity is unknown, but it may represent precursor forms of NT.

Amidated joining peptide in the human pituitary, gut, adrenal gland and bronchial carcinoids. Immunocytochemical and immunochemical evidence.

The distribution of the proopiomelanocortin-derivated amidated joining peptide (JP-N) was examined in the human pituitary gland, adrenal gland, gut and in three bronchial carcinoids. Double immunostaining showed coexistence of immunoreactive JP-N and other proopiomelanocortin derivatives, e.g., ACTH, beta-endorphin, Pro-tau-MSH, in the pituitary gland and adrenal medulla. The JP-N immunoreactive cells in the adrenal medulla were identified as a subpopulation of adrenaline-producing cells by means of an antiserum against phenylethanolamine N-methyltransferase. In the gut immunoreactive JP-N was costored with somatostatin in endocrine cells. Using radioimmunoassay, JP-N was found in higher concentrations than ACTH and alpha-MSH in the gut but not in the adrenal gland. Gel chromatography of gastric antrum and adrenal gland extracts showed three and two dominating components of immunoreactive JP-N, respectively, but under reduced conditions most of the immunoreactive material appeared as of low molecular weight in both extracts. In conclusion, immunoreactive JP-N is a major product from the processing of proopiomelanocortin in human extrapituitary tissues. The molecular forms of immunoreactive JP-N correspond to previous findings in the human pituitary gland.

Characterisation by mass spectrometry and 500-MHz proton nuclear magnetic resonance spectroscopy of penta- and hexasaccharide chains of human foetal gastrointestinal mucins (meconium glycoproteins).

Structural studies using liquid secondary ion mass spectrometry, gas liquid chromatography/mass spectrometry and 500-MHz 1H NMR are described of the major penta- and hexasaccharides of a fraction of human foetal gastrointestinal mucins. Glycoproteins from a blood group H active meconium pool were studied after depletion of Ii antigenic activities by immunoaffinity chromatography and treatment with mild acid hydrolysis to reduce the chain heterogeneity. Oligosaccharides were released by mild alkali/borohydride degradation and purified by Bio-Gel P4 chromatography and HPLC. Eleven penta- and hexasaccharides have been fully characterised as a result of this study and one previous report [Hounsell et al. (1988) Biochem. J. 256, 397-401] and information obtained on additional oligosaccharides present in small amounts. These oligosaccharides show the following features: (table; see text) Sequences in these oligosaccharides not commonly found in mucins so far studied are chain-terminating GlcNAc alpha 1-4Gal, repeating-type-I (Gal beta 1-3GlcNAc) backbones, the backbone branch GlcNAc beta 1-6(GlcNAc beta 1-3)Gal and the backbone sequence GlcNAc beta 1-6Gal beta 1- in the absence of a substituent at C3 of galactose.

The distribution, purification, and pharmacological action of an amphibian neuromedin U.

The distribution, primary structure, and relative biological activity of neuromedin U has been determined from the frog Rana temporaria. Following sequential column chromatography of a gastrointestinal extract, the peptide was sufficiently pure to enable characterization by micro-sequence analysis. The entire sequence was found to be an icosapentapeptide which displays marked sequence similarity to both porcine and rat neuromedin U. The sequence of the biologically active, COOH-terminal region is almost completely conserved across all species. Synthetic, COOH-terminally amidated amphibian neuromedin U, like the porcine and rat peptides, stimulates rat uterine contraction in vitro thereby fulfilling the criterion upon which the nomenclature of this peptide family is based. In addition, the peptide demonstrates parallel pressor effects when infused systemically into rats. The high degree of amino acid sequence conservation is indicative of strong evolutionary pressure acting to retain the presence of this possibly physiologically important peptide across the vertebrate subphylum.

[Cellular localization, half-life, and secretion of peptide YY]. / Localización celular, vida media y secreción del péptido YY.

Tissue and plasma concentration of peptide YY (PYY) were measured by means of a radioimmunoassay (RIA) developed in our laboratory, using a specific PYY antiserum generated in New Zealand white rabbits against synthetic PYY, and dextran-coated charcoal to terminate the assay. Cellular localization of PYY was studied immunohistochemically using the peroxidase-antiperoxidase (PAP) technique. The highest tissue concentration of PYY was found in the mucosa of the terminal ileum and colon. PYY-containing secretory granules were primarily found in the basal pole of open-type endocrine cells. Basal plasma concentration of PYY was 70 +/- 9 pg/ml and rose to 357 +/- 30 pg/ml during the IV administration of PYY at 400 pmol/kg-h. A significant correlation was found (r = 0.94, p less than 0.05) between dose of PYY (12.5, 25, 50, 100, 200, 400 pmol/kg-h, IV) and plasma concentration of PYY. The calculated half-life of PYY in plasma was 8.3 +/- 1.9 minutes. Plasma concentration of PYY during the intraduodenal administration of sodium oleate (150 +/- 20 pg/ml) or long-chain triglyceride (187 +/- 37 pg/ml) was similar to plasma concentration of PYY obtained during the IV administration of PYY at 100 pmol/kg-h. Plasma concentration of PYY raised (126 +/- 10 pg/ml) after the administration of bombesin (400 pmol/kg-h, IV). Bile enhanced release of PYY. The present study suggests a hormonal role for PYY.

Histological and histochemical studies on the digestive gland of Bulinus truncatus infected with Schistosoma haematobium.

The structure of the uninfected and infected digestive gland of the fresh-water snail, Bulinus truncatus, was studied histologically and histochemically. The digestive gland consists of two types of cells: digestive and secretory cells clearly differentiated from each other. The pathological effects of infection of the snail with cercariae of Schistosoma haematobium have resulted in considerable structural and histochemical alterations in the digestive gland. Generally, the main effects were reflected by increased cellular vacuolation and marked diminution of carbohydrates, proteins and lipids of the gland cells.

Quantitation and characterization of peptides from the C-terminal flanking region of rat and bovine preprotachykinins.

Sequence analysis of cDNAs has shown that the biosynthetic precursors of substance P (alpha-, beta-, and gamma-preprotachykinins) contain a common amino acid sequence in the C-terminal flanking region that has not been conserved between species. Antisera have been raised against the synthetic peptide Tyr-Glu-Arg-Ser-Ala-Met-Gln-Asn-Tyr-Glu, which represents rat beta-preprotachykinin-(117-126)-peptide, and used in radioimmunoassays. Antiserum R50 reacted strongly with C-flanking peptides in extracts of rat and bovine tissues whereas antiserum GP-4 reacted only with the rat peptides. The primary structure of the predominant molecular form of preprotachykinin C-flanking peptide in an extract of bovine corpus striatum was established as: Ala-Leu-Asn-Ser-Val5-Ala-Tyr-Glu-Arg-Ser10-Val-Met-Gln-Asp-Tyr1 5-Glu. This sequence represents beta-preprotachykinin-(111-126)-peptide which is equivalent to gamma-preprotachykinin-(96-111)-peptide. A C-flanking peptide with similar chromatographic properties was identified in extracts of rat brain and gut together with a second immunoreactive component that may represent a fragment or a posttranslationally modified variant. A peptide corresponding to the 37-amino-acid residue C-flanking peptide derived from alpha-preprotachykinin was not detected in the extracts as expected from the known low abundance of alpha-preprotachykinin mRNA in rat brain and gut.

Amyloidosis in pigtailed macaques (Macaca nemestrina): pathologic aspects.

The pathologic aspects of 248 cases of amyloidosis in pigtailed macaques (Macaca nemestrina) at the Washington Regional Primate Research Center from 1971 through 1985 were studied. Amyloid was present in the spleen, liver and gastrointestinal (GI) tract, either alone or in combination, in nearly 75% of the monkeys. Its occurrence declined with age in the spleen and the GI tract, but increased with age in the liver. Both intestinal inflammation and retroperitoneal fibromatosis were strongly associated with amyloid deposition in the GI tract. Monkeys with histopathologic findings of enteritis or enterocolitis and glomerulonephritis were at increased risk of developing amyloidosis. Forty cases of amyloidosis with a history of chronic diarrhea had type AA amyloid by histochemical tests.

Immunohistochemical localization of chromogranin A in normal tissues from laboratory animals.

To analyze the distribution of Chromogranin A in endocrine cells of various species of laboratory animals (dog, gerbil, guinea pig, hamster, monkey, mouse, and fetal, neonatal, and adult rats), normal tissues were stained immunohistochemically with polyclonal anti-bovine Chromogranin A antiserum (SP-1). Selected tissues (pituitary, adrenal, thyroid, parathyroid, pancreas, brain, peripheral nerve, stomach, small and large intestine, bone marrow, spleen, thymus, lymph node, and liver) from these species and from the rabbit were stained with two monoclonal anti-human Chromogranin A antibodies (LK2H10 and PHE5) to compare the immunoreactivities of the monoclonal antibodies and polyclonal antiserum. Staining with the polyclonal antiserum (SP-1) resulted in a broader spectrum of immunoreactivity but had more nonspecific background staining than either monoclonal antibody. Immunoreactivity and staining intensity with SP-1 varied between species, but most endocrine tissues (pituitary cells in the anterior and intermediate lobes, thyroid "C" cells, adrenal medulla, parathyroid, pancreatic islets, and enterochromaffin cells) from most species stained positively. In some species, pancreatic alpha cells stained more intensely, and two populations of adrenal medullary cells with different staining intensities were observed. Sciatic nerve (axonal area) was immunoreactive with monoclonal antibodies and/or the polyclonal antiserum in several species. The spectrum of immunoreactive tissues from fetal and neonatal rats increased with age. There was good cross-reactivity between species with SP-1, but not with either LK2H10 or PHE5. These results indicate that many endocrine cells with secretory granules in laboratory animals express Chromogranin A and that a polyclonal antiserum, such as SP-1, is more sensitive in detecting this protein in various species than monoclonal antibodies such as LK2H10 or PHE5.

[Detection of gut hormone mRNAs by synthetic DNA probes in gut hormone producing tissues].

Synthetic DNA probes were tested as hybridization probes for detecting gut hormone mRNAs. When 4 gut hormones, including gastrin, somatostatin, gastrin-releasing peptide and calcitonin were examined, these probes were shown to be useful for mRNA detection in the tissues producing respective hormones. It was also revealed that there was a good correlation between the concentration of peptides determined by radioimmunoassay and the amounts of mRNAs. This methodology was applied for multiple gut hormone producing tumor with the aim to elucidate the mechanisms responsible for this phenomenon, and demonstrated that the tumor expressed a large amount of mature mRNAs encoding respective hormones. These results indicate that increase of mRNA production is one of the mechanism responsible for multiple gut hormones production by tumor.

Aluminum speciation studies in biological fluids. Part 2. Quantitative investigation of aluminum-citrate complexes and appraisal of their potential significance in vivo.

Because of the recent implications of aluminum in the pathogenesis of various disease states, its in vivo chemistry has been receiving growing attention from bioinorganic chemists over the last few years. In this context, the elucidation of the main factors that govern aluminum bioavailability constitutes an urgent objective. Clearly, prevention measures require that mechanisms of aluminum absorption be definitely characterized, whereas specific sequestering agents are needed to detoxify patients with high-aluminum-body burdens. In particular, speciation studies are necessary to discriminate among the chemical forms under which aluminum predominates in vivo. Low molecular weight (LMW) species, which are the most active in terms of bioavailability, cannot be assessed by analytical techniques, and so computer simulations must be used. In recent clinical studies as well as in preliminary simulations dealing with aluminum distribution in blood plasma, citrate has been recognized as the most important LMW ligand of aluminum. The present paper thus reports a quantitative investigation of aluminum-citrate equilibria, carried out at 37 degrees C in NaCl 0.15 mol dm-3 in accordance with the experimental protocol defined in our previous study on aluminum hydrolysis. The ML, MLH, ML2, M3L3H-4, M2L2H-2, ML2H-1, and ML2H-2 species have been characterized over the whole physiological pH range using as large reactant concentration ratios as possible. Corresponding formation constants have then been used to investigate the role of citrate towards aluminum bioavailability. Blood plasma simulations reveal that citrate can promote aluminum urinary excretion, which substantiates recent clinical observations made on mice. However, the higher plasma aluminum concentrations are, the less effective citrate is to be expected. Gastrointestinal simulations confirm that the electrically neutral ML complex does represent an important risk of aluminum absorption in the upper region of the gastrointestinal tract at usual therapeutic doses. At moderate- and low-aluminum concentrations, citrate is also capable of dissolving the aluminum trihydroxide precipitate, which may combine with the capacity of other ligands to complex Al3+ into absorbable complexes at less acidic pH.

Hepatic and renal extraction of circulating type III procollagen amino-terminal propeptide and hyaluronan in pig.

Hepatic and renal clearance of the amino-terminal propeptide of type III procollagen (PIIINP) and of the glycosaminoglycan, hyaluronan (HA) were investigated in a catheterization study of seven healthy anesthetized pigs. Two assays were used, in order to distinguish between the metabolism of different PIIINP-related antigens. One was the PIIINP RIA Kit, which measures the intact propeptide. The other was the PIIINP Fab assay, in which the antibody has an equal affinity to the intact propeptide and to smaller fragments, of which the latter constitutes most of the antigenic activity in serum. Hepatic and gastrointestinal extraction were evaluated from measurements of serum concentrations in the artery, the portal vein and the hepatic vein. We found a significant hepatic extraction of the intact propeptide (extraction ratio 0.14) and of HA (extraction ratio 0.23), but not of smaller PIIINP fragments. No gastrointestinal extraction of any of the tested substances could be demonstrated. Only smaller PIIINP fragments (such as the col 1 fragment) were extracted by the kidneys (the extraction ratio in the PIIINP Fab assay was 0.19). The renal extraction ratio of HA was 0.14. The amounts of PIIINP fragments and of HA extracted by the kidneys were 50- and 3-times the amounts found in urine, respectively, indicating that the col 1 fragment and HA are degraded in the kidneys in addition to urinary excretion. Our results suggest a dynamic turnover of connective tissue-related components with a fast catabolism of circulating components in liver and kidneys.

Distribution of acid stable trypsin inhibitor immunoreactivity in normal and malignant human tissues.

The distribution and localization of acid stable trypsin inhibitor (ASTI) in normal and malignant human tissues from various organs were examined using immunohistochemical techniques that used goat antibody raised against highly purified ASTI from human urine. Tissues were assessed as positive only when they were stained by both the biotin-avidin-peroxidase complex system and biotin-streptavidin-beta-galactosidase complex system, and the staining was abolished by absorption with purified ASTI. Under normal conditions, ASTI immunoreactivity was observed in only a few organs. Positive tissues for ASTI immunoreactivity included the kidney proximal tubules, glial cells of the cerebrum, fibrillar structures of the lamina propria of the stomach and colon, and bronchial epithelial cells. No ASTI immunoreactivity was observed in the cardiovascular system, reproductive system, or other tissues examined. As is not the case for normal tissues, ASTI immunoreactivity was found to be widely distributed in malignant tumors. Staining was observed in the extracellular space, i.e., in the stroma of the tumor and in connective tissues around the tumor invasion, whereas no ASTI immunoreactivity was detected in the malignant cells. Considering the identity of the first 36 NH2-terminal residues of ASTI purified from plasma or urine with a recently reported endothelial cell growth factor, the present findings suggest that ASTI could play an important role, not limited to its function as a protease inhibitor, in the invasive growth of malignant neoplasms.

Expression of the rat amylin (IAPP/DAP) gene.

We have used the polymerase chain reaction with mixed sequence primers to generate a probe for rat amylin and have used this to detect expression in various rat tissues. Amylin mRNA is found in greatest concentrations in the pancreas where a single mRNA species can be detected giving a hybridisation signal intensity approximately 10% that of insulin mRNA. When the beta cell population was depleted with streptozotocin, both amylin and insulin mRNAs were reduced to a similar extent. Consistent with its supposed role in the control of carbohydrate metabolism, amylin mRNA was also found in the stomach. Unlike the related peptide, CGRP, amylin mRNA is not present in the thyroid and is not widely distributed in the central nervous system. The only nervous tissue in which it could be detected was the dorsal root ganglion. Surprisingly, amylin mRNA was also found in the lung though only at very low levels.

Autoradiographic localization of sigma opioid receptors in the gastrointestinal tract of the guinea pig.

The distribution of sigma and phencyclindine binding sites in the gastrointestinal tract of the guinea pig was studied by autoradiography after in vitro incubation of tissue slices with (+)3H-SKF 10,047 and 3H-1-1-[(2-thienyl)cyclohexyl] piperidine to locate sigma and phencyclidine receptors, respectively. A dense distribution of sigma binding sites was observed in the mucosa and in the submucosal plexus, particularly at the level of the fundus and duodenum. Muscular layers were only marginally labeled. No phencyclidine binding site could be demonstrated in any area. This selective distribution suggests a functional role of sigma receptors mainly in the control of endocrine or exocrine secretions, or both.

Time course of expression of neuropeptide Y, calcitonin gene-related peptide, and NADPH diaphorase activity in neurons of the developing murine bowel and the appearance of 5-hydroxytryptamine in mucosal enterochromaffin cells.

Serotoninergic and cholinergic neurons are known to appear earlier in the ontogeny (day E12) of the murine gut than those containing substance P or vasoactive intestinal peptide (day E14). It has also been demonstrated that proliferating neural precursors coexist with mature neurons in developing enteric ganglia. These observations have led to the hypotheses that peptidergic neurons develop later than those that utilize small molecule neurotransmitters and that the activity of early developing neurons may affect the phenotypic expression of coexisting neuroblasts. As a partial test of these hypotheses we studied the phenotypic expression of neurons recognized by antisera to neuropeptide Y (NPY) and calcitonin gene-related peptide (CGRP), and of those visualized by the histochemical demonstration of reduced nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase activity. NADPH diaphorase activity, which is coexpressed with NPY immunoreactivity in all submucosal and many myenteric neurons, was first found on day E11 in clusters of cells in the dorsal mesogastrium. These cells also expressed neurofilament reactivity and thus were developing along a neuronal lineage. Enteric neurons that expressed NADPH diaphorase activity were visualized in the stomach one day later, on day E12. At this time, NADPH diaphorase-containing cells could no longer be demonstrated in the dorsal mesogastrium. NPY immunoreactivity first appeared in the wall of the bowel on day E12, when it was seen in cells in the presumptive stomach. By day E13, the entire length of the bowel contained NPY-immunoreactive neurons. Cells that displayed NADPH diaphorase activity were found at this time at both ends of the alimentary tract, but did not appear in the ileum until day E18. In contrast, CGRP immunoreactivity could not be detected anywhere in the gut until day E17, but by day E18 all regions of the bowel contained CGRP-immunoreactive neurons. Endogenous 5-HT was first detected at day E16 in mucosal epithelial cells in all segments of the gut except the stomach, where it appeared at day E18. The NPY/NADPH diaphorase set of neurons thus develop before the acquisition of a detectable level of endogenous 5-HT or enteric neural 5-HT receptors (which arise in the foregut at day E14). These observations demonstrate that enteric neurons that express small molecule neurotransmitters do not necessarily develop earlier than peptidergic neurons as a class; however, various types of enteric neurons do appear in a sequential order.(ABSTRACT TRUNCATED AT 400 WORDS)