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.

Metabolism of benzo[a]pyrene on the nasal mucosa of Syrian hamsters: comparison to metabolism by other extrahepatic tissues and possible role of nasally produced metabolites in carcinogenesis.

The formation of metabolites of nasally instilled benzo[a]pyrene [(BP) CAS: 50-32-8] was determined. The study was prompted by a report that a high incidence of tumors occurred in tissues of the upper respiratory and alimentary tracts of Syrian hamsters exposed to BP aerosols. The esophagi of anesthetized hamsters were surgically catheterized so that radiolabeled material instilled as BP in the nose could be collected and analyzed for metabolites. About 50% of the instilled BP was metabolized in the nose and, potentially, would have been swallowed in an awake animal. In auxiliary experiments, homogenates of respiratory and alimentary tissues were tested for metabolic activity for BP. The nose, trachea, and lungs had about equally high activities on a per organ basis (5-7 nmol/hr), whereas all other tissues had considerably less activity. The results of the study indicate that nasal metabolism may be important in causing tumors in alimentary as well as upper respiratory tissues.

Distribution of peptide histidine isoleucine in the mammalian respiratory tract and some aspects of its pharmacology.

Peptide histidine isoleucine (PHI), a newly discovered neuropeptide, has been detected by RIA and immunocytochemistry in the upper respiratory tracts of the guinea pig, rat, and cat. HPLC of tracheal extracts showed a single peak of PHI immunoreactivity in each species. The immunoreactive PHI peak found in the guinea pig and rat trachea was eluted earlier than the corresponding peak from the cat, which was coeluted with the porcine PHI standard. Immunocytochemistry showed PHI immunoreactivity to be present within ganglion cells and nerve fibers in the respiratory tracts of all three species. The distribution of PHI was similar to that of vasoactive intestinal polypeptide and ganglion cells were found to contain both PHI and vasoactive intestinal polypeptide immunoreactivities. Pure natural porcine PHI induced a dose-dependent relaxation of isolated guinea pig tracheal muscle which was not blocked by antagonists to catecholamines, histamine, 5-hydroxy-tryptamine, and acetylcholine. PHI may thus be one of the local factors in respiratory control.

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.

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.

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.

Studies on the distribution of PHI in mammals.

We have developed a sensitive and specific radioimmunoassay to PHI and investigated its distribution in four mammalian species (man, cat, guinea-pig and rat). PHI was present in high concentrations, not only in intestine but also in brain, respiratory tract, urogenital tract and other peripheral tissues. Its distribution was similar to that of VIP and in each tissue examined there was always a significant correlation between the concentrations of these two peptides. In a survey of endocrine tumours, PHI was found to be produced only in those tumours that also produced VIP. In addition PHI was only elevated in the plasma of patients that also had high plasma VIP concentrations. This parallel distribution and release was found to be due to the co-synthesis of VIP and PHI in the same pro-hormone peptide. However, the variable ratio of VIP/PHI in different anatomical areas suggest that in these areas there is a different post-translational enzyme processing of the precursor protein.

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.

Inhalation toxicity studies on cigarette smoke III. Tobacco smoke inhalation dosimetry study on rats.

Smoke inhalation dosimetry studies have been carried out on rats, using a new exposure system. A range of cigarettes, tobacco types and smoke concentrations was used. Penetration of smoke into the lungs was clearly demonstrated, and loads of total particulate matter (TPM) of up 1 mg were detected in the lower respiratory system of rats. The mass of TPM deposited was affected by the smoke concentration during exposure. Deposition of TPM in the head of the rat was low in relation to total respiratory system deposition. A pattern of predominantly lung deposition was achieved under the conditions used for this series of experiments. This pattern was not affected by changes in smoke dilution level, cigarette or tobacco type.

Inhalation toxicity studies on cigarette smoke II. Tobacco smoke inhalation dosimetry studies on small laboratory animals.

A newly developed exposure system has been used to carry out smoke dosimetry studies on rats, mice, hamsters and guinea pigs. In all species, smoke total particulate matter (TPM) deposited in significant amounts in the lower respiratory system (LRS) at dose levels ranging from 0.515 to 1.710 mg TPM/g respiratory tissue. Nasal deposition of smoke particulates did occur in all of the species examined. The significance of these dosimetry data in relation to the conduct of long-term comparative inhalation toxicity studies with tobacco smoke is discussed.

Study of biological samples obtained from victims of MGM Grand Hotel fire.

Eighty blood samples and 17 respiratory-tract tissue samples containing fluid taken from victims of the MGM Grand Hotel fire were studied to assist in the determination of the cause of death. The blood and tissue-fluid samples were analyzed for carboxyhemoglobin, oxyhemoglobin, methemoglobin, and total hemoglobin. Outgassing studies were done on the tissue samples using gas chromatography/mass spectroscopy, and heavy metal analysis on inhaled soot was done by x-ray fluorescence. The carboxyhemoglobin values obtained on the samples were significantly higher than those reported by Clark County. However, the percentage of the victims with a carboxyhemoglobin saturation level of 50% or less is higher than that found in the Maryland fire fatality study, suggesting that other toxic factors may have contributed to the lethal nature of the fire.

Tissue distribution of amyloid deposits in Abyssinian cats with familial amyloidosis.

The tissue distribution of amyloid deposits was studied in 15 related Abyssinian cats with familial amyloidosis. There was interstitial medullary amyloidosis in the kidneys of all 15 cats but only 11 had detectable glomerular involvement. The thyroid glands, stomach and colon were affected in all cats examined. Most of the cats also had amyloid deposits in the small intestine, spleen, heart, adrenals, pancreas, liver, lymph nodes and bladder. In 50 per cent or fewer of the cats examined, there was involvement of the parathyroids, lung and gonads. The central nervous system was not involved in any of the 3 cats evaluated. In 8 of the cats, no concurrent inflammatory disease could be detected. The tissue distribution of amyloid deposits resembled that found in other breeds of domestic cats with systemic amyloidosis. Despite the wide tissue distribution of amyloid deposits, clinical signs were related to renal amyloidosis. Familial amyloidosis in the Abyssinian cat may represent a valuable spontaneous animal model for the study of Familial Mediterranean Fever in man and the pathogenesis of reactive amyloidosis in general.

The U.S. Transuranium Registry report of the 241Am content of a whole body. Part IV: Preparation and analysis of the tissues and bones.

Los Alamos National Laboratory has analyzed autopsy tissue for the USTR, as a part of its study of the uptake, distribution and retention of Pu and other transuranic elements in occupationally exposed workers since 1978. In April 1979, Los Alamos received the internal organs and bone samples from the first whole-body donation to the USTR. The donor was known to have an internal deposition of 241Am. All soft tissue, the bones from the right half of the skeleton, and the odd-numbered vertebrae were received at Los Alamos in February 1980. The bones were subdivided along anatomical areas of interest. All soft tissues and bone specimens were analyzed for their 241Am content. A total deposition of 147.4 nCi 241Am was measured. Approximately 18% of the 241Am remaining in the body (disregarding that in the left hand), was found in the soft tissues, and 82% was in the bones and teeth. The soft tissues and organs containing the largest amounts of 241Am were the combined soft tissue (striated muscle, connective tissue and skin) 8.8%; liver, 6.4% and respiratory tract, 1.5%. The remaining organs accounted for 0.9% of the systemic burden.

[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.

[Investigations of the pathogenesis of lung cancer observed among chromate factory workers].

In order to clarify the relationship between the chromate compound and occurrence of lung cancer, the author studied the characteristics of patients with lung cancer among workers of chromate factory and measured the chromium contents of each tissues obtained from 14 patients at surgery and autopsy. The incidence of the chromate lung cancer was 413 per 100,000 population, which was 16 times that of the general population. All were male. The age ranged from 26 to 74 year old (average 53). The histological type was mostly squamous and small cell carcinoma. Location of carcinoma occurrence was mainly limited to the large bronchi. The average total labor period of patients with lung cancer was 258 months and the latent period was 305 months. The working history of the patients in hexavalent-chromium producing process was longer than that of the control group. Patients with small cell carcinoma was mainly engaged in the monochromate producing process. The labor and latent period of patients with squamous cell carcinoma was longer than those of small cell carcinoma. Measurement of chromium contents in the respiratory system of chromate workers revealed much higher chromium content than in control group. The chromium content of tissues in the non-respiratory system was a little higher than that of the control group. High chromium content itself did not have any relation with the occurrence of lung cancer because the primary location of chromate lung cancer was limited to the large bronchi, not to the peripheral lungs which contained the highest chromium content. The longer was the exposure period, the higher was chromium content in the lung. Chromium content in the upper lobe was higher than that in the lower lobe. From these studies, the author concluded that hexavalent-chromate compound might be the compound responsible for lung cancer occurrence.