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.

Histochemical characterization of glycoconjugates in the epithelium of the extrapulmonary airways of several vertebrates.

The glycoconjugates of the extrapulmonary airways of 11 tetrapode vertebrates have been characterized by means of both conventional and lectin histochemistry. Abundant sialosulphomucins were detected in the secretory cells and periciliary layer of turtles, snakes, birds and mammals while only sialomucins were observed in amphibians. Neutral and traces of acidic mucins were detected in the secretory cells of lizards. The secretory cells of the amphibian airways were reactive to Con-A, DBA and WGA. No alpha-L-fucose residues reactive with UEA-I or LTA were detected in amphibians. The goblet cells of the turtles were stained by DBA, SBA and WGA. Secretory cells of snakes and lizards reacted with Con-A and WGA. The mucous goblet cells of the birds were reactive to Con-A, LTA and WGA. In the chicken, they also showed affinity for PNA and SBA. The ciliated cells of the avian species studied were stained by Con-A and WGA. Mammalian goblet cells were reactive to Con-A, UEA-I and WGA. In the rat, affinity for DBA and SBA was also observed. The present results reveal the existence of marked differences in the sugar residues of the glycoconjugates of the extrapulmonary airways of tetrapode vertebrates. Only sialic acid residues appear to be constant constituents of the glycoconjugates of the airways of all species studied.

[Radioimmunoassay of vasoactive intestinal peptide immunoreactivity in guinea pig respiratory tract].

Although vasoactive intestinal peptide (VIP) has been suggested to be the neurotransmitter of non-adrenergic and non-cholinergic (NANC) inhibitory nerves, its physiological functions and movements in the airway are obscure. In this study, VIP immunoreactivity in the respiratory tract from guinea pigs was measured as a preliminary experiment to elucidate its functions. VIP immunoreactivity was measured by radioimmunoassay. The rate of VIP disappearance during extraction was 52.5 +/- 17.4 (mean +/- SD)%. The dose-response curve of tissue extract almost paralleled the standard curve of authentic VIP. VIP immunoreactivity of tracheas was 939.9 +/- 262.1 pg/g wet weight and that of extrapulmonary bronchi was 858.0 +/- 241.1 pg/g wet weight. Although VIP immunoreactivity of lung extracts was not detectable in 14 samples out of 23, the value of 9 samples was 111.7 +/- 61.5 pg/g wet weight. These results suggest that there may be more VIP immunoreactivity present in tracheas and extrapulmonary bronchi than in lungs.

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.

[Using specific radioimmunoassay for guinea pig VIP (GP VIP), immunoreactive GP VIP levels in respiratory tract tissue of normal and airway-allergic guinea pigs].

Vasoactive intestinal polypeptide (VIP) has recently received widespread attention with respect to its control of airway constriction, and is now regarded as the most promising candidate as a neurotransmitter for nonadrenergic noncholinergic inhibitory nerves in the airway. A specific system of radioimmunoassay (RIA) was devised to measure guinea pig VIP (gp VIP) concentration in airway tissue. An antiserum, R 550, was obtained by immunizing rabbits with newly synthesized gp VIP. The detectable VIP concentration was assessed to be 10 fmol/tube with the RIA system. The concentration of gp VIP in the respiratory tract tissue of normal guinea pigs was 0.04-0.22 pmol/g wet weight of tissue. The VIP level was highest in the trachea, in contrast to lower levels in both the major bronchus and lower lung. After being passively sensitized with ovalbumin, the experimental guinea pigs were exposed to the same antigen. After three minutes, as the expiration time was prolonged and the respiratory resistance increased, the immunoreactivity in the trachea and the major bronchus was significantly higher than the pre-exposure levels. After six hours, however, they returned to the original levels except in the lower lung where the level was constant throughout the respiratory change. As described above, the possibility of controlling airway constriction by VIPergic nerve in immediate-type allergic reactions is suggested by our findings.

Non-adrenergic non-cholinergic nervous control of airways.

Traditionally, the tone of bronchial smooth muscle is mediated through the balance of two autonomic nervous systems. The existence of a third non-adrenergic non-cholinergic (NANC) nervous system in the lung has been demonstrated in animals. In contrast to high density parasympathetic innervation of smooth muscle, no evidence of adrenergic nerves was found in human airways. Electrical field stimulation of tracheal strips in guinea-pig produced an initial contraction followed by a relaxation. The contraction was blocked by atropine but the relaxation was not inhibited by a beta-adrenergic blocking agent. Immunohistochemical studies have shown that vasoactive intestinal peptide (VIP)-like immunoreactive nerves are present in the central nervous system and in the peripheral neuronal system of mammals including the human respiratory tract. Autoradiographic and immunocytochemical studies have demonstrated a high density of VIP receptors in airway epithelium, submucosal glands, pulmonary vessels and smooth muscle, especially in large airways. Functional studies in humans have confirmed the regulating role of VIP principally in the large airway.

Immunocytochemical localization of epidermal growth factor in the developing human respiratory system and in acute and chronic lung disease in the neonate.

Cells staining for immunoreactive human epidermal growth factor were sought in the lungs and tracheas of human fetuses from 8 to 24 weeks of gestation. Lungs of liveborn infants from 25 to 40 weeks of gestation (stillborn to 7 months postnatal life), both with and without lung pathology, were also studied. In the early fetal trachea (12 to 15 weeks), many nonciliated cells immunostained for immunoreactive human epidermal growth factor in the lining epithelium. By 16 weeks of gestation this widespread staining was replaced by stained nonciliated single cells or small clusters of cells which were identifiable until 24 weeks. In the few tracheas which were available from liveborn infants who died without evidence of lung disease, stained cells were seldom identified in the lining epithelium after 24 weeks of gestation. In contrast, from 18 weeks until term, tracheal submucosal glands contained scattered cells which immunostained for immunoreactive human epidermal growth factor but which did not appear to be classical mucous cells. Beginning at 20 weeks of gestation, positively staining cells were found occasionally in bronchial lining epithelium, but more often in bronchial submucosal glands. Immunostained cells were never identified in bronchiolar epithelium in normal fetal or newborn lungs. In liveborn infants from 24 weeks onward who developed lung disease, many tracheas were severely damaged. In the presence of extensive denudation of the mucosa or the development of squamous metaplasia, immunostained cells were rarely seen in the lining epithelium. However, even under these conditions stained glandular cells could usually be identified. Stained cells were also present in the necks of those tracheal glands from which new epithelial lining cells appeared to be migrating onto denuded surfaces. Immunostained cells in the bronchial lining epithelium of infants with chronic lung disease were infrequent, just as they were in the fetus, but bronchial submucosal glands contained positively stained cells similar to those in tracheal glands. The appearance and distribution of immunostained cells were similar in the tracheal and bronchial submucosal glands in both normal subjects and those with all stages of lung disease. In contrast to the bronchioles of fetuses and infants without lung pathology, the bronchiolar epithelium of infants with chronic lung disease contained immunostained cells. Immunostained cells were found in areas of migrating dysplastic cells in relining conducting airways.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of pulmonary oxygen injury on airway content of surfactant-associated protein A.

The use of therapeutic hyperoxia has greatly improved the survival of infants born prematurely. However, high concentrations of oxygen cause pulmonary injury, leading to decreased pulmonary compliance and decreased oxygen diffusion. This injury can result in chronic pulmonary insufficiency. It has been hypothesized that the adverse effects of hyperoxia are mediated, in part, through changes in the pulmonary surfactant system. We investigated the effects of hyperoxia on surfactant-associated protein A (SP-A), the abundant surfactant-specific glycoprotein. Adult male rats were exposed to 85% oxygen for 72 h. Total lung volume and pulmonary compliance were measured, and alveolar surfactant material recovered by lavage. Hyperoxia decreased total lung capacity, and altered inflation and deflation hysteresis patterns. Disaturated phosphatidylcholine and SP-A content were significantly increased in alveolar surfactant material isolated from oxygen-treated rats. SP-A content was also significantly increased in lung tissue from oxygen-treated rats. The SP-A in the lavage of oxygen-treated rats appeared to be intact protein as no proteolytic fragments were detected and the SP-A migrated identically to that recovered from room air animals when analyzed by two-dimensional isoelectric focusing. We conclude that the decreased pulmonary compliance associated with pulmonary oxygen injury is not due to quantitative decreases in two major surfactant components, disaturated phosphatidylcholine and SP-A.

Spiramycin concentrations in the human respiratory tract: a review.

Spiramycin has exceptionally good distribution properties, especially in respiratory tract tissues and fluids. Three hours after a single oral dose of 3 g spiramycin, the serum concentration ranged from 1.6 to 2.8 mg/l and the reported half-life was approximately 8 h. Studies of lung tissue concentrations showed that high pulmonary levels were achieved after a loading dose of 3 g; the levels were higher after multiple doses and reached approximately 30 to 45 mg/kg in lung tissue and 6.5 to 36 mg/kg in bronchial mucosa; in bronchial secretions and in sputum the concentrations of spiramycin ranged from 1.5 to 7.3 mg/l (after 1 g, multiple doses). In upper respiratory tract tissues and fluids, high levels of spiramycin were reached as well: 8 to 13 mg/kg in sinus mucosa; 15 to 29.5 mg/kg in tonsils or adenoids.

Occurrence and distribution of glycoconjugates in human tissues as detected by the Erythrina cristagalli lectin.

We applied a horseradish peroxidase-Erythrina cristagalli agglutinin (HRP-ECA) conjugate for histochemical staining of tissue sections from various formalin-fixed, paraffin-embedded human tissue specimens. The HRP-ECA conjugate showed broad reactivity, but there was a distinct distribution of native (not masked by sialic acid) and sialic acid-masked ECA binding sites in the various organs. Free ECA binding sites could be detected on red blood cells, lymphocytes of thymus, tonsil, lymph node, and in mucous substances of different organs. Independent of blood group type, the vascular endothelium exhibited strong ECA reactivity. Free ECA binding sites occurred in the cytoplasm of Kupffer's cells in liver, in histiocytic cells of thymus, lymph node, tonsil, and in bone marrow. Podocytes of kidney glomerulus, syncytiotrophoblasts of placenta, megakaryocytes in bone marrow, myelin sheath of nerve, medullary thymocytes, and hepatocytes, as well as islet cells of pancreas, contained only sialic acid-capped ECA binding sites. Inhibiting studies with galactose, lactose, and N-acetyl-lactosamine, as well as other sugars, revealed that this lectin is specific for galactosyl residues. In comparison to galactose and lactose, N-acetyl-lactosamine exhibited the highest inhibitory activity on lectin binding, supporting the concept that this lectin is most reactive with N-acetyl-lactosamine-type (type 2 chain) glycoconjugates.

Immunoglobulin G and its function in the human respiratory tract.

Immunoglobulin G--which can be subdivided into four classes, each with different functional characteristics--is an important component of the host defense system of the respiratory tract. An excessive amount can be produced or can accumulate after airway irritation (exposure to cigarette smoke) or from immunologic stimulus of B-lymphocyte-plasma cells in types of hypersensitivity and interstitial lung diseases. Specific antibody activity can be identified in organic dust-induced hypersensitivity pneumonitis and asthma that contributes to disease pathogenesis. The availability of opsonic antimicrobial antibodies is essential for optimal function of phagocytes in uptake and containment of bacteria. With an absolute or functional deficiency of IgG, recurrent and chronic types of sinopulmonary infections occur. These extremes of IgG availability, either high levels (presumably excessive) or deficient, are discussed in this review.

Anatomical distribution of vasoactive intestinal peptide binding sites in peripheral tissues investigated by in vitro autoradiography.

Vasoactive intestinal polypeptide has a widespread distribution in the body, occurring in both the central and peripheral nervous systems and considerable information is available on its distribution, physiology, and pharmacological actions. Receptors for VIP have been demonstrated previously in peripheral tissues by conventional binding techniques using isolated membrane preparations. However, information on their precise localization is limited. We therefore localized binding sites in a variety of guinea pig and rat tissues by in vitro autoradiography and made a parallel study of the distribution of VIP nerves in these tissues using immunocytochemistry. [125I]VIP was prepared by the chloramine T method and shown to be pharmacologically active. After a preincubation procedure to remove endogenously bound VIP, unfixed cryostat sections were incubated with 1 nM [125I]VIP. To determine specific binding, sections were incubated in the presence or absence of 1 microM unlabeled VIP. Autoradiograms were generated by exposing the sections to LKB-Ultrofilm or emulsion-coated coverslips. Dense binding occurred in discrete locations within the gastrointestinal, respiratory, and genital tracts, correlating with known actions of VIP and, to various extents, with the distribution of VIP nerves. For example, there was precise localization to respiratory epithelium, smooth muscle of airways and blood vessels, and alveolar walls, in keeping with the effects of VIP on vascular and airway smooth muscle and mucus secretion.

Structure and mass of mammalian respiratory ciliary outer arm 19S dynein.

Mammalian respiratory ciliary outer arm dyneins isolated as the major ATPase peak migrating at 19S on sucrose density gradients were examined by transmission electron microscopy of negatively stained samples and scanning transmission electron microscopy of unstained samples. The predominant discrete particle structure observed was composed of two globular heads apparently connected by amorphous or indistinct material. The heads were either circular or slightly elliptical of mean 13 +/- 1 X 10 +/- 2 nm dimensions. The mass of this structure averaged 1.22 +/- 0.34 million daltons with the individual globular heads averaging 310 +/- 77 kilodaltons (kD). Negative staining revealed that one or both of the globular heads often contained a central accumulation of stain measuring 2.5 +/- 1 nm across. A second type of structure, appearing with lesser frequency in the 19S fraction than in the unfractionated dynein preparation loaded onto the sucrose gradient, was a single globular head of 13 +/- 1 X 10 +/- 2 nm often with 2 +/- 1 nm centrally accumulated stain and with or without an appendage. This one-headed particle thus resembled one-half of the two-headed particle. Mass measurements were lower, however, for isolated, single globular heads, averaging 220 +/- 111 kD. A third type of particle observed was a ring-like structure with 4 +/- 1 nm centrally accumulated stain and without appendages. The ring structure was slightly larger in diameter, 14 +/- 1 nm, and had a greater peripheral accumulation of negative stain than either of the one- or two-headed particles, suggesting that it was not derived therefrom.(ABSTRACT TRUNCATED AT 250 WORDS)

Regulatory peptides in the respiratory system.

Many regulatory peptides have been described in the respiratory tract of animals and humans. Some peptides (bombesin, calcitonin, calcitonin gene-related peptide) are localised to neuroendocrine cells and may have a trophic or transmitter role. Others are localised to motor nerves. Vasoactive intestinal peptide and peptide histidine isoleucine are candidates for neurotransmitters of non-adrenergic inhibitory fibres and may be cotransmitters in cholinergic nerves. These peptides may regulate airway smooth muscle tone, bronchial blood flow and airway secretions. Sensory neuropeptides (substance P, neurokinin A and B, calcitonin gene-related peptide) may contract airway smooth muscle, stimulate mucus secretion and regulate bronchial blood flow and microvascular permeability. If released by an axon reflex mechanism these peptides may be involved in the pathogenesis of asthma. Other peptides, such as galanin and neuropeptide Y, are also present but their function is not yet known.

Regulatory peptides in the respiratory tract of Macaca fascicularis.

The quantitative distribution and localisation of seven regulatory peptides (vasoactive intestinal peptide (VIP), peptide histidine methionine (PHM), calcitonin gene related peptide (CGRP), galanin, substance P, neuropeptide tyrosine (Y), and bombesin like peptides) were determined by radioimmunoassay and immunocytochemistry in six different regions of the respiratory tract of the cynomolgus monkey, Macaca fascicularis. In general, peptide concentrations were higher in the airways than in lung tissue itself. VIP and PHM were found in greatest abundance and in equimolar concentrations. Concentrations of substance P, neuropeptide Y, and bombesin were substantially lower. Immunocytochemistry localised all the peptides to nerve fibres, whose density generally paralleled the tissue concentrations by radioimmunoassay except in the case of bombesin, which was not detected. VIP, PHM, and galanin were mostly associated with glands of trachea and bronchus and with blood vessels and smooth muscle; CGRP and substance P were found principally beneath airway epithelium and around smooth muscle fibres and blood vessels; neuropeptide Y was found around blood vessels and seromucous glands only. The pattern of peptide distribution in the Macaca fascicularis respiratory tract is similar to that previously reported in human postmortem material, suggesting that the cynomolgus monkey may be a useful model for examining the pathophysiological role of peptides in human respiratory disease.

Comparison of dust samplers: statistical analysis techniques.

Statistical analyses comparing pairs of dust samplers were carried out using five models, a straight line through the origin, linear with intercepts, logarithmic, a logarithmic (weight + constant), and a fifth forced through the origin. The fourth and fifth models combined the arithmetic portion of the error, associated with weighing and handling, with the geometric portion, associated with sampling and the dust cloud variations, into a unified geometric form. It was shown that some of the assumptions on which the least squares method of regression analysis are based were not met and that corrections had to be made. In most cases no significant difference was found between the corrected regression lines and the expected physical relation, a straight line through the origin. In the exceptions physical reasons were found to support the nonlinearity found in statistical analysis. It is suggested that the best estimate of the relation between two dust samplers can be obtained by a least squares determination of the straight line through the origin using transformed variables. A method is given for determining values of the constants. The application of this statistical method to comparisons between dust samplers not measuring the same parameter of the dust cloud is discussed with an example. The considerations in assessing historical dust measurements are outlined.

The structure and innervation of the saccopleural membrane of the domestic fowl, Gallus gallus: an ultrastructural and immunohistochemical study.

Microscopic studies have shown the saccopleural membrane in the respiratory system of the domestic fowl to consist of a sheet of three dense layers of collagen fibres covered dorsally and ventrally by mainly simple squamous epithelium. On the ventral surface, which faces into the caudal thoracic air sac, there are occasional ridges of pseudostratified ciliated epithelium. Many nerve bundles are present throughout the membrane, the larger bundles of myelinated and unmyelinated axons being confined to the lamina propria under the dorsal epithelium (parietal pleura). In addition to axonal profiles with the ultrastructural appearance of cholinergic or adrenergic axons, peptidergic-type axons were identified. Immunofluorescence studies demonstrated VIP-, substance P-, somatostatin- and enkephalin-immunoreactive fibres in the membrane. Although it has been suggested that receptors may be present in this region of the respiratory system, none of the axons have features suggestive of sensory terminals, although many axonal profiles are closely associated with the epithelia where no obvious effector cells are present.

Isolation of an olfactory cDNA: similarity to retinol-binding protein suggests a role in olfaction.

Molecular cloning techniques were used to isolate and characterize a protein possibly involved in the signal transducing system in olfactory tissue of the frog Rana pipiens. A complementary DNA library was constructed with messenger RNA obtained from frog olfactory neuroepithelium. A 700-base pair complementary DNA clone encoding a protein with a molecular weight of 20,300 was identified by differential hybridization analysis with polyadenylated RNA from olfactory epithelium and nonsensory respiratory epithelium. The messenger RNA corresponding to this clone was abundant in the cells of Bowman's glands in olfactory tissue but not in respiratory epithelium nor in several other tissues. The predicted sequence of this protein is homologous to members of a family of proteins that bind and transport small molecules in serum, suggesting that this protein may also bind and transport odorants in the mucus secreted by Bowman's glands.