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Biomedical subjects

J E Brinn

Publications and source records attributed to J E Brinn.

At least 19 recordsLinked to original sources

Endogenous codeine: autocrine regulator of catecholamine release from chromaffin cells.

In addition to the catecholamines (CAs) dopamine (DA), norepinephrine (NE) and epinephrine (E), perifused chromaffin cells of the eel secrete codeine and morphine. In controls, the release of NE and E is strongly correlated, while there is no correlation with DA. Low, physiological concentrations of codeine (500 pg/ml) reduce the release of NE and E, while 8-fold higher concentrations stimulate an instant, transitory release of all three CAs. Much higher concentration of codeine (100 ng/ml), corresponding to the therapeutically effective range in the human, again reduce the release of NE and E. Physiological and very high concentrations of morphine have no clear effect on CA release, while an intermediate concentration (38 ng/ml) increases the secretion of all three CAs. The opiate antagonist naloxone lowers the basal CA secretion and prevents the morphine-induced CA increase. During naloxone perifusion, a normally non-effective concentration (40 ng/ml) of codeine reduces the CA release. It appears that codeine is an autocrine regulator which suppresses CA release via naloxone-insensitive receptors, and stimulates CA release via opiate receptor(s). Co-released morphine may modulate the action of codeine.

Anguilla↗

Effects of combined secretagogues and extracellular calcium on neonatal insulin release.

The insulin response of 3-day old neonatal rat islets was evaluated following a 1 h incubation with glucose alone and in the presence of 30 nM sulfated cholecystokinin octapeptide (CCK) and/or 20 microM carbachol (CCh). Insulin secretion was found to be incrementally increased from the lowest glucose concentration and enhanced several fold in the presence of CCK and/or CCh. In combination, CCK and CCh increased glucose-stimulated insulin secretion by an amount equivalent to the sum of their individual increases. The presence of either CCK alone or CCK plus CCh increased phosphoinositide hydrolysis by the same relative amount that they increased insulin secretion when compared to 8.3 mM glucose. Glucose-stimulated insulin secretion was totally inhibited when calcium was omitted from the incubation buffer; this effect was partially negated by CCK alone and more so by CCK combined with CCh. Insulin secretion in response to 8.3 mM glucose alone was unchanged when calcium in the incubation buffer was increased from 1 to 5 mM; however, the insulin response to 16.7 mM glucose alone and 8.3 mM glucose in the presence of CCK and/or CCh was increased under this condition. Thus, we have shown that, even at 3 days postpartum, insulin secretion from isolated islets is a complex response capable of being molded by several secretagogues at once and ultimately determined by interplay of different signaling systems activated.

Animals↗

Catecholamine release by catecholamines in the eel does not require the presence of brain or anterior spinal cord.

The catecholamine-producing chromaffin cells of the American eel are strongly innervated by fibers, which, by ultrastructural criteria, seem to be cholinergic. However, neither removal of the brain nor removal of the brain combined with extirpation of the anterior spinal cord prevents the release of catecholamines into the circulation by catecholamines. It appears that the chromaffin cells are controlled by both nervous and humoral stimuli, and that at least some of the latter do not require the presence of "preganglionic" innervation.

Anguilla↗

Age-dependent stimulation of neonatal insulin release and inositol phosphate accumulation by CCK-8 and carbachol.

Development of a robust insulin secretory response to glucose occurs during the early neonatal period. To determine if neuroendocrine agents play a role during this time, we studied the effects of selected peptides and neurotransmitters on insulin release and polyphosphoinositide metabolism in islets isolated from 1- and 3-day neonatal rats. Vasoactive intestinal peptide had no effect on glucose-stimulated release in either islet population. In contrast, sulfated cholecystokinin octapeptide (CCK-8) significantly enhanced glucose-induced insulin release in both islet groups. One-day islets were stimulated only by a concentration of 300 nM, whereas 3-day islets were responsive at 3 nM. Similar to CCK-8, there were clear differences in responses to carbachol between 1- and 3-day islets. One-day islets required a concentration of 200 microM for insulin release to be significantly greater than with glucose alone; 3-day islet insulin release was significant at 2 microM carbachol. Both agonists stimulated inositol phosphate accumulation in 3-day islets, but only CCK-8 caused a significant increase over glucose-induced levels in 1-day islets. These results indicate that islet responsiveness to CCK-8 and carbachol develops in parallel during the early neonatal period. This development may be linked to the maturation of a critical step of stimulus-secretion coupling through which these agents act.

Age Factors↗

Mongolian gerbil tympanic membrane. Normal and with induced otitis media.

Otoscopic observation of the tympanic membrane is the most effective noninvasive means of diagnosing and following the course of experimental otitis media in animal models of the disease in both short-term and in longitudinal studies. Because of the difficulty of viewing the entire tympanic membrane of the gerbil model of otitis media without manipulation, we present methods of visualizing and color illustrations of the normal tympanic membrane of the Mongolian gerbil, Meriones unguiculatus. These are compared with illustrations of animals with acute experimental otitis media. An artist's rendition of the entire normal gerbil tympanic membrane is presented. These methods and illustrations will enable investigators to more rapidly and easily adopt the Mongolian gerbil as an animal model for studies of otitis media.

Acute Disease↗

Vasoactive intestinal polypeptide enhances hormone content and insulin release in cultured fetal rat islets.

The effect of porcine vasoactive intestinal polypeptide (VIP) on development of the biphasic insulin release response in cultured fetal rat islets was investigated. Fetal islets, 21.5 days gestational age, were cultured for 7 days in RPMI 1640 culture medium containing either 2.8 or 11.1 mM glucose adn subsequently challenged with 16.7 mM glucose in a perfusion system. Islets were exposed to VIP at a final concentration of 13.2 nM by adding the peptide to the perifusion buffer (acute exposure) or by adding it to the culture medium throughout the culture period (chronic exposure). Islet hormone and DNA contents were also quantitated at the end of the culture period. Acute exposure to VIP resulted in no alterations of the insulin release pattern after culture in the presence of either glucose concentration. However, chronic treatment of islets with 13.2 nM VIP in the presence of 2.8 mM glucose resulted in significant increases in the maximum rate of insulin release during the first phase and the total amount of insulin release during both phases. Similarly, islets cultured in the presence of 11.1 mM glucose and 13.2 nM VIP demonstrated enhanced biphasic insulin release patterns with increased maximum rate and total amount of release during both phases. The presence of VIP and 2.8 mM glucose increased islet glucagon and somatostatin contents, but islet DNA and insulin contents remained unchanged. These findings indicate that VIP plays a significant role in the in vitro development of the biphasic insulin release pattern and may be a factor controlling the maturation of the fetal islet in vivo.

Animals↗

Longitudinal studies of experimental otitis media with Haemophilus influenzae in the gerbil.

Haemophilus influenzae non-typable strain 119 was found to cause severe otitis media with sequellae when inoculated into the middle ear cavities of the Mongolian gerbil, Meriones unguiculatus. Acute inflammation was followed by the development of highly vascular granulation tissue and formation of new bone within the middle ear bulla. These changes persisted throughout the 14-week study in the untreated animals. The gerbil was variably susceptible to otitis media caused by inoculating 30-3000 bacterial cells and 100% susceptible to greater than 3000 cells. The susceptibility of the gerbil to common etiological agents of otitis media allows its use as an appropriate model of the disease.

Acute Disease↗

Experimental pneumococcal otitis media: longitudinal studies in the gerbil model.

In order to ascertain that the Mongolian gerbil, Meriones unguiculatus, is an acceptable model for studying the development of chronic from acute otitis media, we used previously published methods for experimental otitis media in a longitudinal study of the acute disease and sequellae. The gerbil was found to be susceptible to as few as 30 viable Streptococcus pneumoniae type 3 cells, indicating that only a few viable pathogens are able to cause the disease. Untreated experimental infections with S. pneumoniae type 23 resulted in a mild, self-limiting disease with little permanent sequellae, while S. pneumoniae type 3 produced severe disease characterized by an acute phase of from 2 to 3 weeks, followed by the development of new bone formation and a vascularized granulation tissue which persisted throughout the 13-week study. Viable pneumococci could be recovered from the middle ears for only two weeks. We conclude that the gerbil is a useful model for otitis media.

Animals↗

An immunocytochemical study of the pancreatic islet system of the channel catfish.

The endocrine pancreas of the channel catfish is segregated into a large primary islet and numerous smaller secondary islets. In view of cell distribution differences in mammalian islets of ventral and dorsal primordia, we have determined the percentage volumes of insulin-, glucagon-, and somatostatin-containing cells in primary and secondary catfish islets to ascertain if these islets correlated with those derived from ventral and dorsal primordia in mammals. Islets were immunocytochemically stained using antisera to anglerfish insulin, porcine glucagon, and synthetic somatostatin and volume densities were quantified on light micrographs by point-counting procedures. In both primary and secondary islets the insulin-, glucagon-, and somatostatin-containing cells comprised approximately 32%, 23%, and 38% of the endocrine cell volumes, respectively. Therefore, the cell populations did not reflect any embryological differences between the two groups of islets. In this study, the volume densities of insulin-reactive cells in the primary islet were less than previously reported, and the overall insulin staining was about one-half of that seen in mammals. The volume density of somatostatin-reactive cells in primary islets was greater than previously reported. Based on these data, primary and secondary islets of the catfish do not appear to have a similar development to the ventral and dorsal islets of the mammalian system.

Animals↗

Glucose affects in vitro maturation of fetal rat islets.

Fetal pancreatic islets (21.5 days old) were cultured in RPMI 1640 containing either 2.8 or 11.1 mM glucose for 7 days. After the 7-day culture period, islets cultured in 2.8 mM glucose demonstrated a minimal first phase of insulin secretion in response to acute glucose stimulation, whereas islets cultured in 11.1 mM glucose demonstrated a biphasic insulin secretory pattern. Islets cultured in 11.1 mM glucose initiated insulin secretion at 4.4 +/- 0.1 mM glucose and plateaued at 11.6 mM glucose when exposed to a linear gradient. In addition, culture in 11.1 mM glucose increased DNA content (P less than 0.01) and [3H]thymidine incorporation (P less than 0.05) in fetal islets. However, ultrastructural morphometric analysis indicated that the actual number of beta-cells within islets cultured in either 2.8 or 11.1 mM glucose did not increase. The insulin contents of islets cultured in 2.8 and 11.1 mM glucose were 0.46 +/- 0.06 and 1.14 +/- 0.10 mU/islet, respectively. During subsequent glucose stimulation, islets cultured in 2.8 and 11.1 mM glucose released 3% and 5.6% of their total insulin content, respectively. Ultrastructural morphometric analysis indicated that 11.1 mM glucose stimulated an increase in the volume of individual beta-cells, i.e. hypertrophy. The hypertrophy of beta-cells within islets cultured in 11.1 mM glucose resulted in a concomitant increase in islet volume. Finally, the hypertrophy of beta-cells within islets cultured in 11.1 mM glucose was a result of increased volumes of mitochondria, secretory granules, and, to the greatest extent, endoplasmic reticulum. These findings indicate that glucose is a potent factor in the maturation of cultured fetal rat islets.

Animals↗

Effects of growth hormone on the in vitro maturation of fetal islets.

To study the effects of growth hormone (GH) on the in vitro maturation of fetal islets, the fetal islets were cultured for 7 days in RPMI 1640 containing 10% fetal bovine serum and 11.1 mM glucose with or without GH. Culture with 1 microgram/ml of bovine GH increased the DNA content of the islets and [3H]thymidine incorporation into DNA confirming results of other investigators. In addition, however, the insulin secretory dynamics and ultrastructural morphometrics were investigated. It was found that GH-treated islets demonstrated increased insulin release during acute glucose stimulation when expressed as microunits per islet per minute. However, when insulin release during acute glucose stimulation was expressed as microunits per microgram of DNA per minute to compensate for the increased DNA content of GH-treated islets, no change in insulin release was observed compared to control islets. When GH-treated islets were perifused with a linear glucose gradient, the insulin secretory response was suppressed as indicated by changes in the threshold level, plateau level, and half-maximal response. Ultrastructural morphometric data showed that the average beta-cell volume in control and GH-treated islets was the same, eliminating the possibility that beta-cell hypertrophy occurred. Similarly, the nuclear volumes of the beta cells in control and GH-treated islets remained unchanged. This finding coupled with the observed increased DNA content and [3H]thymidine incorporation suggests that GH functions by increasing cell multiplication within the islets and not by inducing polyploidy. Finally, the volumes of cytoplasmic organelles in control and GH-treated islets were the same indicating that cytodifferentiation did not occur.

Animals↗

Experimental otitis media in gerbils and chinchillas with Streptococcus pneumoniae, Haemophilus influenzae, and other aerobic and anaerobic bacteria.

To ascertain the usefulness of Mongolian gerbils as an inbred model for otitis media, 52 Mongolian gerbils (Meriones unguiculatus, strain MONT/Tum) were compared with 26 chinchillas (Chinchilla laniger) for susceptibility to Streptococcus pneumoniae type 3. Haemophilus influenzae type b, and a polymicrobic culture including anaerobes (Streptococcus intermedius, Propionibacterium acnes, Staphylococcus epidermidis, and Corynebacterium sp.). Organisms were inoculated percutaneously into the superior chamber of the middle ear bulla. The gerbils and chinchillas shared similar susceptibilities and responses to the inoculated organisms as determined by X-ray, otoscopic, histopathological, and microbiological determinations at 5 to 7 days. Koch's postulate studies proved the role of S. pneumoniae and H. influenzae in the pathology found in both animal models. The animals were also susceptible to the polymicrobic culture, although the relative virulence of the individual members of this mixture was low, suggesting that these species potentiated as a polymicrobic mixture. The Corynebacterium sp. appeared to elicit the greatest histopathological response in chronic (8-week) studies in gerbils. The gerbils were found to be useful as an alternative animal model for the study of otitis media of bacterial etiology.

Animals↗

Innervation of the cultured fetal rat pancreas.

The fetal rat pancreas, explanted at 18 days of gestation and cultured up to ten days, contains numerous acetylcholinesterase-positive neurons. These nerves usually appear in small ganglia although single nerve cells are encountered. The axons of these intrapancreatic nerves appear to terminate only in the islet tissue and not on any exocrine components of the explant. It is concluded that the fetal rat pancreas contains an islet-specific group of cholinergic neurons.

Animals↗

New types of islet cells in a cyclostome, Petromyzon marinus L.

Four types of acidophilic granular cells, in addition to B-cells, are identified in the islet organ of anadromous specimens of two subspecies of Petromyzon marinus by light and electron microscopy. Three of these acidophils (PI, PII and PIV-cells) occur in both the cranial and hepatic islets while a fourth type (PIII-cell) has only been found in the hepatic islet of some animals. The granules of the PI-cells stain with ponceau de xylidine, give a distinct tryptophan reaction and in ultrastructural examination show large, dense granules. The PII-cells contain unusual crystals and appear to be a non-secretory stage of the PI. The PIII-cells stain deep-red and acid fuchsin. They contain very large, dense granules and some lysosomes. PIV-cells stain selectively with phosphotungstic acid-hematoxylin and ultrastructurally, contain small, more or less dense granules. It appears that PI- and PIV-cells develop directly from B-cells, while the PIII-cells derive from PI-cells. despite their direct or indirect origin from B-cells, the PI-, PIII- and PIV-cells show characteristic features of functionally independent endocrine cells. Petromyzon marinus may be an ideal model for the understanding of phylogenetic and pathological interrelationships between islet and gastrointestinal hormones. It is clear that the interpretation of the islet organ of the cyclostomes, which has been generally considered a source of insulin only, requires a revaluation.

Animals↗

Pancreatic islet cytology of Ictaluridac (Teleostei).

The endocrine pancreas of the bullhead catfish, Ictalurus nebulosus, and the channel catfish, I. punctatas was studied by light and electron microscopy. In addition to the usual A, B and D cells, a fourth endocrine cell type was consistently observed in the electron microscope. All endocrine cell types were innervated. The vesicles of most of the nerve endings were ultrastructurally different from typical adrenergic and cholinergic vesicles, strongly suggesting the possibility of a third autonomic neurotransmitter serving as a regulator of catfish islet secretion.

Animals↗

Stains for A, B, and D cells in fetal rat islets.

Ten techniques often used for identification of A, B, and D cells in adult islets of Langerhans were applied to fetal rat pancreas. Modifications were tried with many of these techniques. Two indole methods (xanthydrol and postocoupled benxylidene reactions) and a cryostat technique using o-phthaladehyde failed to stain fetal islets. Phosphotungstic acid hematoxylin and lead hematoxylin lightly stained fetal A cell granules in Helly's fixed tissue. The Grimelius silver nitrate technique stains adult rat A cells but failed to stain fetal cells. A modification of this technique stained fetal A cells and a possible 4th cell type. The specificity of this method was confirmed by restaining stained cells with a fluorescent antibody technique and with pseudoisocyanin. B cells, as previously reported, were readily stained by the aldehyde fuchsin technique. Fetal D cells were not stained by the Hellerstrom-Hellman alcoholic silver nitrate method, nor did they display pseudoisocyanin metachromasia after acid hydrolysis; they did fluoresce brightly with this technique when viewed with UV light. It was thus possible to distinguish the three usual cell types, plus a possible fourth type, in the fetal rat pancreas.

Animals↗