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Biomedical subjects

C Yanaihara

Publications and source records attributed to C Yanaihara.

At least 19 recordsLinked to original sources

[Structure-function studies of galanin].

Galanin is widely distributed in the central and peripheral nervous system and exerts a variety of physiological effects. This review briefly describes the chemical structure, tissue distribution, physiological effects, receptors and structure-function relationships of galanin. It is worth noting that the inhibitory effect of newly synthesized galanin (1-15)-ol on guinea pig ileum contractions was of the same magnitude as that of galanin. This observation gives us an important clue as to the discovery of antagonists of galanin for neural systems.

Amino Acid Sequence

Galanin-induced alteration of electrolyte transport in the rat intestine.

Effects of rat and porcine galanin on rat intestinal ion transport were examined in vitro. In the rat distal colon, a sustained increase in short-circuit current (Isc) was produced by the serosal addition of rat galanin at a concentration as low as 10(-9) M, and a maximal increment was observed at 10(-7) M. Porcine galanin was approximately 100 times less potent than rat galanin. In the rat jejunum, rat galanin produced only a slight and transient decrease in basal Isc. The response to rat galanin was not influenced by atropine, hexamethonium, or amiloride, but was virtually abolished by tetrodotoxin or furosemide. Rat galanin did not significantly influence the increase in Isc elicited by electrical field stimulation in the rat colon and jejunum. Transmural unidirectional 22Na and 36Cl fluxes in the rat colonic mucosa were measured under short-circuited conditions, and rat galanin significantly decreased net sodium and net chloride absorption. These findings suggest that galanin acts as a secretory modulator in the rat colon via noncholinergic neural transmission.

Amiloride

[Drug compliance in the elderly].

In order to clarify the characteristics of elderly patients concerning their attitudes toward taking prescribed medicine, self-reported compliance with prescriptions was compared among different age groups. We performed a survey in 626 outpatients and their attending physicians in 4 of our affiliated hospitals, and analyzed self-reported compliance by the patients to the prescription and their answers to questions related to drug-taking along with the diagnoses and prescriptions reported by the physician. The number of prescribed medicine was 2.3 tablets on the average for patients younger than 40, while 5.1 tablets were prescribed for patients over 70. However, self-reported compliance was best in patients over 70 than in other age groups. Compliance was good in 76% of patients who answered that the amount of medicine was appropriate, while good compliance was lower in those who thought the prescription excessive (67%). Likewise, compliance of patients who had concerns with drug side effects or who were not feeling well under medication was lower than that of patients who felt well under medication. Prescriptions for after lunch were most liable to be forgotten than those for other times of the day. Moreover, a high percentage of elderly patients attended more than 2 departments or medical facilities, and one third of those patients did not inform the physician of the fact suggesting that they were at higher risks of overdosing and unexpected drug interaction. Furthermore, the percentage of patients who did not receive explanation from physicians was higher in the elderly thus demonstrating that elderly patients have quite different characteristics in attitude with regard to medicine from younger age group patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Ultrastructural identification of human secretin cells by the immunogold technique. Their costorage of chromogranin A and serotonin.

We have localized secretin in a morphologically distinctive endocrine cell scattered in the epithelium covering the villi and uppermost crypts of the human duodenum and jejunum. The human secretin cell was characterized by relatively large (mean diameter 299 nm +/- 69 SD), fairly irregular granules, the majority of which showed homogeneous distribution of secretin and chromogranin A immunolabelling in a structurally homogeneous core. Other granules had a targetoid pattern due to an inner, argyrophobe, secretin-immunoreactive body surrounded by an argyrophil, chromogranin A immunoreactive mantle. These targetoid granules represent a distinctive ultrastructural marker of the secretin cell. Secretin cell granules have been shown to react with chromogranin A antibodies and Grimelius' silver, while lacking chromogranin B immunoreactivity. About 1/3 of secretin cells also showed serotonin immunostaining.

APUD Cells

Content and distribution of vasoactive intestinal polypeptide (VIP) in cavernous tissue of human penis.

Penile erection is controlled by a valvular structure in the helicine artery in humans. The opening and closing of this valve are believed to be regulated by the autonomic nervous system, especially through the release of vasoactive intestinal polypeptide (VIP). We determined the content of VIP in cavernous tissue in 18 impotent patients and in 5 normal controls by radioimmunoassay, and we examined the distribution of VIP-ergic nerve fibers in cavernous tissue by an immunohistochemical method. As a result, it was found that the lower penile VIP content was more frequent among patients with organic impotence than among the controls. Furthermore, VIP-ergic nerve fibers were seen to be diffusely and loosely distributed in a large number of organic impotence patients. These findings suggest that organic impotence in some patients may be due to decreases in the VIP content and in VIP-ergic nerve fibers.

Adolescent

Epidermal growth factor and its receptors in human pancreatic carcinoma.

The role of epidermal growth factor (EGF) in oncogenesis and progression of malignant tumors is a subject of vast interest. In this study, radioimmunoassay and radioreceptor assay of EGF were established. EGF contents in malignant and benign pancreatic tumors, in normal pancreas tissue, and in culture media of a human pancreatic carcinoma cell line were determined. EGF receptor binding studies were performed. It was shown that EGF contents in pancreatic carcinomas were significantly higher than those in normal pancreas or benign pancreatic tumors. EGF was also detected in the culture medium of a pancreatic carcinoma cell line. The binding of 125I-EGF to the pancreatic carcinoma cells was time and temperature dependent, reversible, competitive, and specific. Scatchard analysis showed that the dissociation constant of EGF receptor was 2.1 X 10(-9) M, number of binding sites was 1.3 X 10(5) cell. These results indicate that there is an over-expression of EGF/EGF receptors in pancreatic carcinomas, and that an autocrine regulatory mechanism may exist in the growth-promoting effect of EGF on tumor cells.

Epidermal Growth Factor

Gonadotropin-releasing hormone-associated peptide immunoreactivity in bovine colostrum.

Gonadotropin-releasing hormone(GnRH)-associated peptide (GAP) is a 56-amino acid peptide found on the C-terminal of the GnRH (also called luteinizing hormone-releasing hormone) precursor and is assumed to be co-produced with GnRH. The purpose of this report is to demonstrate the presence of GAP immunoreactivity in bovine colostrum. Radioimmunoassay of acidified methanolic extracts demonstrated a concentration of GAP immunoreactivity of approximately 1.5 +/- 0.1 pmol/g dry skim bovine colostrum. Gel filtration (Sephadex G-10) and high-performance liquid chromatography of extracts containing GAP immunoreactivity showed it to be of low molecular weight and a high hydrophobic character. The presence of GAP immunoreactivity in bovine colostrum suggests that the GnRH precursor is synthesized and processed in mammary tissue itself.

Animals

Immunocytochemical analysis of [Met5]enkephalin-Arg6-Gly7-Leu8 immunoreactive structures in the rat superior cervical ganglion.

Indirect immunofluorescence and immunoelectron microscopy were employed to analyze the enkephalinergic systems in the rat superior cervical ganglion (SCG). These systems were identified using specific antiserum against [Met5]Enkephalin-Arg6-Gly7-Leu8 (ENK-8), a peptide which is derived only from proenkephalin A. Abundant ENK-8 like immunoreactive (ENK-8-LI) neurons and fibers were observed in the SCG, but their distribution patterns were heterogenous; ENK-8-LI neurons were localized preferentially in the caudal two-thirds of the SCG, while immunoreactive fibers were found to be distributed more densely in the rostral one-third than in the remaining part of the SCG. Most of the ENK-8-LI neurons were large and had ultrastructural features resembling those of principal cells, some were identified electron microscopically as small intensely fluorescent (SIF) cells. ENK-8-LI fibers were varicose in appearance and surrounded the perikarya of neurons. Since most of these fibers were not detected after experimental decentralization of the SCG and since ENK-8-LI terminals were seen to contain small lucent vesicles, most of the former were thought to be preganglionic fibers. Immunoreactive fibers mainly formed synaptic contacts with the dendrites of non-immunoreactive principal cells, but a small proportion of ENK-8-LI principal cells also received synaptic input from them. Occasionally, immunoreactive fibers formed synapses with the processes or the soma of both ENK-8-LI and non-immunoreactive SIF cells. On the basis of these findings, we conclude that: (1) preganglionic ENK-8-LI fibers terminate mainly on the principal cells, which are devoid of ENK-8-LI structures; (2) the majority of ENK-8-LI neurons are principal cells, while the remainder are SIF cells; (3) inputs to these cells mainly involve structures lacking ENK-8 immunoreactivity; and (4) there are, however, a small number of ENK-8-LI preganglionic fibers which terminate on ENK-8-LI principal cells and SIF cells.

Animals

Proenkephalin opioid peptide product in the sensory ganglia of the rat: a developmental immunohistochemical study.

The distribution and development of Met-enkephalin-Arg6-Gly7-Leu8 (Enk-8)-containing neurons in the sensory ganglia of the rat were investigated by means of immunocytochemistry using specific antiserum to this octapeptide. Enk-8-like immunoreactivity first appeared in neurons of the trigeminal ganglia of the 18-day embryo, then in the dorsal root ganglia of the 21-day embryo, thus exhibiting a rostrocaudal gradient in terms of appearance and abundance. The number of immunoreactive neurons in these sensory ganglia peaked on the 5th-7th postnatal days, with several small ones observed in each section (1.0-1.4% of total cell number). About 30-40% of these Enk-8-like immunoreactive neurons were also immunoreactive to substance P. Subsequently, Enk-8-like immunoreactivity in the sensory ganglia was decreased and was rarely detected in adult animals. However, colchicine treatment revealed the presence of several Enk-8-containing neurons per section prepared from mature rat. All these neurons were small (12.5-25 microns; mean +/- S.E.M., 19.86 +/- 3.26 microns). Some of these were also immunoreactive to substance P. These results strongly suggest that the preproenkephalin A system exists in subpopulations of both developing and matured sensory cells in the rat. Functional significance of this is discussed.

Aging

Effects of growth hormone releasing factor on pancreatic secretion in vivo and in vitro.

Growth hormone releasing factor (GRF), a 44-residue peptide originally isolated from human pancreatic tumors, shows structural similarities to the members of the secretin-vasoactive intestinal peptide (VIP) peptides. This study was designed to determine the effects of human GRF (hGRF-(1-44] on pancreatic secretion in vivo in conscious dogs and in vitro in dispersed rat pancreatic acini. GRF given i.v. in graded doses in dogs caused a small but significant stimulation of pancreatic HCO3- and protein outputs and potentiated secretin- and cholecystokinin (CCK)-induced pancreatic HCO3- but not protein secretion. When given together with somatostatin, GRF failed to reverse the inhibitory action of this peptide on HCO3- and protein responses to secretin plus CCK in dogs. Studies in vitro dispersed rat pancreatic acini showed that GRF added to the incubation medium of these acini caused an increase in basal amylase release and shifted to the left the amylase dose-response curve to caerulein and urecholine but failed to affect the amylase response to VIP. This study indicates that GRF in vivo stimulates basal and augments secretin- or CCK-induced pancreatic HCO3- secretion and that this is probably due to direct stimulatory action of the peptide on pancreatic secretory cells.

Amylases

IGF-II-like immunoreactivity in human tissues, neuroendocrine tumors, and PC12 cells.

We looked for cells that reacted with monoclonal antibodies against rat insulin-like growth factor II (IGF-II). We found them in human fetal kidneys in the 14th week of gestation and in adult adrenal medullas but not in human liver, pancreas, pituitary gland, or thymus. Every pheochromocytoma in a group of 20 had IGF-II-like immunoreactive cells, as did three out of four ganglioneuroblastomas (one of which was heterotransplantable in athymic nude mice) and one carotid body tumor. Using the electron microscope we localized the immunoreactivity in pheochromocytoma cells to their neurosecretory granules of the non-catecholamine type. We failed to find any IGF-II-reactive cells among the pancreatic islet cell tumors, neuroblastomas, medullary carcinomas of the thyroid, retinoblastomas, and Wilms' tumor we studied. One line of human neuroblastoma cells did show some immunoreactivity after treatment with dbcAMP. Additionally, the rat pheochromocytoma cell line PC12 and its subline PC12h occasionally produced a few immunostained cells. These results strongly indicate that human IGF-II is primarily produced in paraganglionic tissues and tumors.

Adrenal Gland Neoplasms

Change in solubility of insulin in therapeutic mixtures of insulin preparations: HPLC analysis.

We systematically and quantitatively observed the changes in solubility when short- and intermediate-acting insulin preparations were mixed together, using high-performance liquid chromatography (HPLC) and centrifugation. We tested 30 combinations of 16 kinds of commercial insulin preparations, including six short-acting and eight intermediate-acting preparations. The soluble insulin component in the short-acting preparations examined was in all cases insolubilized to various extents after mixing. Particularly remarkable was the complete precipitation of the insulin from Actrapid Human when it was mixed with Monotard Human (1:1). This sort of drastic change in insulin solubility after mixture was reflected in the changes in blood glucose level after the subcutaneous injection of the mixture to rabbits. The information provided by pharmaceutical companies on absorption kinetics and action profiles of short-acting insulin preparations may no longer be valid when mixtures with longer-acting preparations are used.

Animals

Demonstration of two distinct insulin-binding components in solubilized human placental membranes by radioimmunoassay using synthetic peptide and anti-synthetic peptide antibody.

A peptide corresponding to the 957-980 sequence of human placental insulin receptor precursor (HIRP) was synthesized and antisera were produced against the synthetic peptide. Anti-synthetic HIRP(957-980) serum HIR-27 was proved to cross-react with HIRP-related proteins in solubilized human placental membranes. A radioimmunoassay developed with the antiserum and synthetic peptide HIRP(957-980) enabled us to separate, in combination with gel filtration, two insulin-binding components in solubilized human placental membranes which conceivably correspond to the alpha 2 beta 2 and alpha beta structures of the placental insulin receptor. The two components were shown to be distinct in insulin-binding behavior depending on conditions of pH and ionic strength in the binding assay.

Antibodies

Comparison of CCK-8 receptors in the pancreas and brain of rats using CCK-8 analogues.

The characteristics of cholecystokinin (CCK) receptors in rat pancreatic acini and in various regions of the brain were examined using synthetic CCK-8 or CCK-7 analogues. 3H-propionylated CCK-8 [( 3H]CCK-8) was used as a ligand. 1) The pancreatic CCK receptor had a single high affinity binding component with a dissociation constant, Kd, of 0.76 nM and a maximum number of specific binding sites, Bmax, of 271.91 fmol/mg protein. On the other hand, the CCK receptor in the cerebral cortex had a Kd of 1.66 nM and a Bmax of 30.15 fmol/mg protein. 2) The order of the potencies of CCK-7 and CCK-8 analogues with a substitution at position 3 or 4 to displace [3H]CCK-8 specific binding to the pancreatic acini was as follows: CCK-8 greater than CCK-7 = SucCCK-7 greater than Suc[Sar3]CCK-7 greater than Suc[D-Trp3]CCK-7 greater than Suc[D-Ala3]CCK-7 greater than [D-Trp4]CCK-8 = [D-Ala4] CCK-8. This order of potencies of CCK analogues was greatly different from that in the cerebral cortex. 3) The carboxy-terminal tetra-peptide (CCK-4) and penta-peptide (CCK-5) had very weak potencies in displacing [3H]CCK-8 binding in the pancreatic acini, which were 20 to 30-fold less than their potencies in the cerebral cortex. These results suggest that the recognition sites for CCK analogues in the pancreatic and brain CCK receptors are different.

Animals

Establishment of the specific radioimmunoassay for serum thymic factor (STF).

Serum thymic factor is a humoral factor involved in the differentiation of T cells. In the present study, a radioimmunoassay system for STF was established using a specific antiserum and an iodinated synthetic STF as a tracer. Serum levels of STF-like immunoreactivities were around 30 pg/ml in human, but in contrast to previous reports serum STF levels did not show age-dependent decreases. STF-like immunoreactivities of the thymic gland were lower than those of the liver and kidney in rats. The livers of human and pig contained high STF-like immunoreactivities. The rat thymic gland extract showed three peaks of STF-like immunoreactivity by gel filtration of Sephadex G-25 which corresponded to the eluted position of authentic STF, a larger molecular size, and a smaller molecular size of STF, respectively, while the rat liver extract showed two peaks corresponding to the void volume fraction and to the position of authentic STF, respectively. When this void volume fraction was digested by trypsin, three peaks which corresponded to authentic STF, a larger molecular size, and a smaller molecular size of STF were observed. Present studies suggest that serum thymic factor is produced as a larger STF molecule in the liver and kidney.

Adult

Recent advances in brain-gut hormones.

Although recent advances in gene technology have made it possible to produce peptides and proteins by the recombinant technique, chemical synthesis either by the solution or by the solid-phase technique is still only a method of choice for the syntheses of a brain-gut hormone and its superagonist and/or antagonist. In addition, synthetic replicates of brain-gut hormones, their analogus and fragments or their precursor-related peptides are important haptenic immunogens to produce region-specific antisera to the respective peptides. In the present study, antisera against proglucagon and pro-LH-RH related peptides were proved to be useful not only for identifying hormone-producing cells, but also for demonstrating the post-translational biosynthetic processing in the cells. For galanin, a novel brain-gut hormone, have structural similarity in its C-terminal region to tachykinins, a monoclonal antibody against synthetic porcine galanin (1-15) was prepared for immunohistochemical study. This monoclonal antibody was found to recognize specifically galanin-producing cells and not to cross-react with any of tachykinins examined. These observations indicate that monoclonal antibodies are highly useful for identifying cells possessing a brain-gut hormone containing structures common to those of different known peptides.

Animals

Presence of epidermal growth factor in human tears.

Epidermal growth factor (EGF) is a polypeptide that stimulates the growth of various tissues, including the cornea. The presence of EGF in tears from normal volunteers and in aqueous humor from cataract patients was investigated via human EGF (hEGF)-specific radioimmunoassay. Immunoreactive hEGF was found to be present at similar concentrations in both reflex (ranging from 0.7 to 8.1 ng/ml) and non-reflex tears (ranging from 1.9 to 9.7 ng/ml), but was undetectable in aqueous humor. Immunoreactive EGF in human tears was indistinguishable immunologically, biologically and biochemically from urine EGF and standard hEGF.

Adult

Co-localization of substance P and Met-enkephalin-Arg6-Gly7-Leu8 in the intraspinal neurons of the rat, with special reference to the neurons in the substantia gelatinosa.

A double-labeling immunofluorescence technique was employed to investigate the co-localization of the functionally antagonistic neuropeptides, substance P and enkephalins, within intraspinal neurons of the rat. Anti-Met-enkephalin-Arg6-Gly7-Leu8 (Enk-8) antiserum was used as a marker of the preproenkephalin A neuron system. The observations were focused on the lumbar spinal cord. Co-localization was most prominent within neurons in the substantia gelatinosa, in which more than 95% of substance P-like immunoreactivity neurons showed Enk-8-like immunoreactivity. These double-labeled cells corresponded to 45% of Enk-8-like immunoreactive neurons in the same area. This suggests that SP/Enk-8 interaction occurs at the axon terminals of the substantia gelatinosa neurons. In deeper layers of the dorsal horn (laminae III, IV), only 14% and 6% of SP-like immunoreactive and Enk-8-like immunoreactive neurons were double labeled, respectively. Co-localization was also observed in neurons located in the laminae I, V, VII and X, suggesting concomitant involvement of these peptides in a variety of spinal cord functions.

Animals