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Biomedical subjects

C Vaillant

Publications and source records attributed to C Vaillant.

At least 55 records · Page 3Linked to original sources

Monolayer and aggregate cultures of rainbow trout hepatocytes: long-term and stable liver-specific expression in aggregates.

An aggregate culture system for rainbow trout hepatocytes was developed to study liver-specific mRNA expression. Maintenance of differentiated functions and morphology of hepatocytes were examined using both monolayer and aggregate systems. The rainbow trout estrogen receptor and vitellogenin genes were induced by estradiol and their mRNAs used as markers of cell differentiation during cell culture. In monolayer culture, rainbow trout hepatocytes established very few cell-cell contacts in minimal media. The use of more complete media promotes cell-cell contacts and cell islet formation. Hepatocyte response to estradiol stimulation was generally lower than in vivo but a correlation between the degree of cellular organization and the intensity of the hormonal response was observed. However, in this system hepatocytes progressively lost their specific hormonal response between 5 and 10 days. In aggregates with DMEM/F12 and Ultroser SF, cell-cell contacts were maximized and stabilized during at least one month. The levels of rainbow trout estrogen receptor and vitellogenin mRNAs induced by estradiol were stable and maintained at a level comparable to in vivo levels; vitellogenin synthesis and secretion remained fully functional for the duration of the culture.

Actins↗

Sequential study of pancreatic structure and function during development of pancreatic acinar atrophy in a German shepherd dog.

Sequential assessments of pancreatic structure and function were performed on a female German Shepherd Dog bred from parents with exocrine pancreatic insufficiency (EPI), to monitor development of pancreatic acinar atrophy in this breed. Determinations of serum trypsin-like immunoreactivity (TLI), results of N-benzoyl-L-tyrosyl-P-aminobenzoic acid test, fecal soy bean stimulation test (SST), and gross and histologic examinations of the pancreas did not provide evidence of exocrine pancreatic disease up to 13 months of age. However, electron microscopy revealed degenerative abnormalities of acinar cells that were already apparent at 6 weeks and became more extensive with age. Examination of the pancreas at 22 months of age also indicated no gross or histologic abnormalities, but electron microscopy revealed widespread degenerative changes, including dilatation of the rough endoplasmic reticulum and extensive fusion of zymogen granules affecting most of the acinar cells. Serum TLI concentration was markedly reduced at that time, indicative of EPI, but the dog remained healthy and results of the SST were normal. Within 1 month, the dog had developed clinical signs of EPI, and not only serum TLI concentration, but also results of the N-benzoyl-L-tyrosyl-p-aminobenzoic acid test and SST were compatible with severe loss of exocrine pancreatic tissue. This loss was confirmed by gross and histologic examination of the pancreas at 25 months, which revealed typical features of pancreatic acinar atrophy, including scattered and disorganized exocrine cells in the small remnants of pancreatic tissue. These findings indicate that in German Shepherd Dogs, pancreatic acinar atrophy may involve interference with normal intracellular processing of zymogen granules, which precedes progressive and eventual rapid loss of exocrine pancreatic tissue.

Aging↗

Metabolic changes during early starvation in rats fed a low-protein diet in the postweaning growth period.

Metabolic changes during the first 24 hours of starvation were studied in rats previously adapted for 3 weeks during the postweaning growth period to a low-protein diet using lactalbumin as a dietary protein source. Previous adaptation to a high-quality, low-protein diet reduced the effects of early starvation on the loss of body and liver weight. In rats fed a low-protein diet (6% lactalbumin, LP rats), free triiodothyronine (T3) concentration remained higher than in control rats (13% lactalbumin, C rats) throughout the experiment (+38%, 24 hours), and the plasma insulin concentration, which was lower than in C rats during the first 6 hours (-56%), was not different thereafter. Plasma insulin to glucagon molar ratio was lower (-54%) and liver cyclic adenosine monophosphate (cAMP) concentration was higher (+28%) in LP than in C rats in the fed state, but these were not different at 24 hours of starvation. Plasma glucose concentration was slightly lower in LP than in C rats (-15%) in the fed state, but it was not different in both groups during starvation. Whereas they were unchanged in the fed state, plasma lactate concentration was lower (-57%) and free fatty acid and total ketone body concentrations were higher (+38% and +183%, respectively) in LP than in C rats at 24 hours of starvation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

In vivo and in vitro studies on sex steroid binding protein (SBP) regulation in rainbow trout (Oncorhynchus mykiss): influence of sex steroid hormones and of factors linked to growth and metabolism.

The respective roles of sex steroids and hormones related to growth and metabolism, on SBP regulation have been studied in rainbow trout. In vivo, oestradiol (E2) supplementation induces a slow but significant increase of plasma SBP concentration. Testosterone or cortisol injections have no effect. In vitro, the steroid binding protein that accumulates in incubation medium of hepatic cell primary cultures has been characterized and found to be similar to blood SBP. Its production is increased by addition of E2 (maximum: +300%). This effect develops slowly over several days of culture and is dose dependent; as little as 1-10 nM E2 is effective. Recombinant rainbow trout GH (rtGH)--0.01 to 1 microgram/ml--also increases SBP accumulation as compared to control cells and seems to maintain SBP production over culture duration. In preliminary experiments, (1) insulin-like growth factor (IGF) and SBP concentrations were found to change inversely after a 4 days stimulation with increasing concentrations of GH; (2) recombinant human IGF1 (250 ng/ml) tended to be inhibitory when SBP production was expressed per mg of total cellular protein, and a micromolar concentration of bovine insulin was clearly inhibitory. Other hormones tested in vitro: triiodothyronine (10-1000 nM), thyroxine (100 nM), 17 alpha, 20 beta-dihydroprogesterone (10-2000 nM), and testosterone (1-1000 nM) did not influence SBP concentration in hepatic cells culture media.

Animals↗

Cellular localization of intrinsic factor in pancreas and stomach of the dog.

A cobalamin (vitamin B12)-binding protein has recently been identified in canine pancreatic juice which is biochemically, immunochemically and functionally similar to canine gastric intrinsic factor. However, the cellular sources of both this pancreatic intrinsic factor and gastric intrinsic factor in the dog are not known. Antisera raised against canine gastric intrinsic factor have been used to examine the distribution of intrinsic factors in the canine pancreas and stomach. Immunoreactivity was demonstrated in duct cells but not acinar or endocrine cells in the pancreas, and in fundic peptic and pyloric gastric pit cells in stomach. All immunostaining was abolished by preabsorption of the antisera with purified canine gastric and pancreatic intrinsic factors. A cellular source of pancreatic intrinsic factor has not been previously described, and the demonstration of intrinsic factor-like immunoreactivity in two cell types in the canine stomach contrasts with its localization in a single cell type in the gastric mucosa of other mammalian species. Furthermore, immunoreactivity in pancreatic duct cells was detected at much higher dilutions of antisera than those required for staining of peptic and gastric pit cells. This suggests a higher concentration of antigen, and supports previous evidence that the pancrease is a major source of intrinsic factor in the dog.

Animals↗

Glucagon-like immunoreactivity in hypothalamic neurons of the rat.

Antisera specific for three different regions of pancreatic proglucagon were used to examine the distribution of such immunoreactivity in rat hypothalamus. Neurons in the supraoptic and paraventricular nuclei were immunoreactive with an antiserum against glucagon, but not with antisera directed towards the aminoterminal region of proglucagon (glicentin) or the glucagon-like peptide I sequence in the carboxyl-terminal region of proglucagon. These findings confirm a previous report of glucagon-like immunoreactivity in the supraoptic and paraventricular nuclei, but indicate that, while this material is immunochemically related to glucagon, it is not derived from a proglucagon-like precursor.

Animals↗

Organometallic derivatives of estradiol as bioligands: targetted binding of the estradiol receptor.

The complexation of estrogens by transitional metal units e.g. (alkyne)Co2(CO)6 and (alkyne)Mo2Cp2(CO)4, at the 17 alpha-position brings about a dramatic change in the chemical behavior of these compounds with respect to that of the free ligands. The 17 beta-OH function becomes particularly labile, even in weakly acidic medium, giving rise to carbenium ion-like species, from which, depending on the metal and the nucleophile, substitution, elimination and rearrangement take place. This situation provides the basis for a new type of active site directed-reagent for estradiol receptor. The hypothesis of vicinal space positioning of an acidic and a nucleophilic group in the estradiol receptor cavity is examined in the light of the amino-acid composition of the steroid binding domain. The requirement of the sulfhydryl group of a cysteine residue is suspected in the first step of the receptor inactivation process.

Animals↗

Identification and estrogen induction of two estrogen receptors (ER) messenger ribonucleic acids in the rainbow trout liver: sequence homology with other ERs.

The estrogen-binding region of the cDNA for chicken ER reveals a mRNA of 3.5 kilobases (kb) in rainbow trout liver. The level of this messenger, which is very low in the liver of naive male animals, can be increased by estrogen stimulation. With this chicken probe, we have isolated a clone from a lambda gt10 trout liver cDNA library. The partial cDNA sequence, which encompasses most of the coding region, shows two domains of striking amino acid homology with human, avian, and Xenopus estrogen receptors (ERs) (DNA binding region: 90%, Hormone binding region: 60%). With this specific probe rainbow trout ER, we detected another messenger (4.5 kb) that is less expressed than the 3.5 kb messenger. The kinetics of stimulation of the two messengers is compared with the kinetics of accumulation of vitellogenin mRNA after E2 administration. This report constitutes the first identification of ER mRNA from a fish.

Amino Acid Sequence↗

Vitellogenin gene expression in primary culture of male rainbow trout hepatocytes.

Using a primary culture of trout hepatocytes we compare the kinetics of accumulation of vitellogenin and its messenger RNA after estrogen administration. We found that the cells were more sensitive to estradiol than to other estrogens. The lowest effective concentration of estradiol was 10(-9) M. At 10(-6) M the androgens have no effect. Comparison of the primary and secondary stimulation with E2 shows that the initial rate of accumulation of vitellogenin is very much higher in the secondary stimulation. Over a time course of primary stimulation we show that after estradiol withdrawal the rate of accumulation of vitellogenin mRNA in the secondary is a function of time of the first stimulation.

Androgens↗

Innervation of facial skin but not masticatory muscles or the tongue by trigeminal primary afferents containing somatostatin in the rat.

Using retrograde transport of a fluorescent dye, True blue, the peripheral tissues innervated by trigeminal ganglion neurones containing somatostatin have been investigated. Of ganglion neurones retrogradely labelled from injections of dye into the facial skin, 3.45% were found to be immunoreactive for somatostatin. In contrast, none of the neurones labelled from injections of dye into the tongue or masseter muscle were found to contain this peptide. This demonstration of a restricted distribution of somatostatin-containing primary afferents raises the possibility that somatostatin may be involved in functions which are specific to skin and not to the other tissues examined.

Animals↗

A social casework contribution to understanding alcoholism.

This article describes a 40-year longitudinal study of alcoholism including the etiology of, progression of, and recovery from the disease. The findings are paralleled with similar studies conducted by social work professionals. The capacity of a longitudinal study to elucidate alcoholism and the recovery process is underscored. The etiological hypothesis that views alcoholism primarily as a symptom of psychological instability is questioned.

Alcoholism↗

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.

Animals↗

Vasoactive intestinal polypeptide stimulation of protein secretion from rat lacrimal gland acini.

The effect of vasoactive intestinal polypeptide (VIP) on protein secretion from lacrimal gland was investigated by using acini prepared by collagenase digestion of rat exorbital lacrimal glands. Protein secretion was determined by incubating the acini for 0-40 min and analyzing the supernatant for peroxidase, a protein secreted by the rat exorbital lacrimal gland. VIP (10(-10) to 10(-7) M) stimulated secretion in a concentration-dependent manner. A maximum concentration of VIP (10(-8) M) stimulated secretion to the same extent as a maximum concentration of carbachol (10(-5) M). The cholinergic antagonist atropine at a concentration (10(-5) M) that completely abolished carbachol-induced secretion did not alter VIP-stimulated secretion. The secretory effects of maximal concentrations of VIP and carbachol were additive, but decreasing the carbachol concentration potentiated secretion. Unlike carbachol, which had no effect on the acinar cAMP level, VIP increased cAMP content sixfold. Immunohistochemical staining demonstrated VIP-like immunoreactivity in nerve fibers throughout the gland, distributed primarily around acini. We conclude that VIP-like immunoreactive nerves are present in the lacrimal gland and that VIP can stimulate protein secretion but utilizes a pathway separate from, but convergent with, that used by cholinergic agonists.

Animals↗

Immunohistochemical studies on the gastrointestinal tract using antisera to Met-enkephalin and Met-enkephalin Arg6Phe7.

Antisera specific for the C-terminus of Met-enkephalin and two of its variants isolated from adrenal medulla and brain, namely Met-enk Arg6 and Met-enk Arg6Phe7, have been used in immunohistochemical studies of the gastrointestinal tract in rat, mouse and guinea pig. Met-enk and Met-enk Arg6Phe7-like immunoreactivities were found with similar distribution in nerve cell bodies of the myenteric plexus, and in fibers that were particularly dense in the myenteric plexus and circular smooth muscle. The Met-enk Arg6 antiserum showed weak staining of nerve cells and fibers. In rat and mouse, the antiserum to Met-enk Arg6Phe7, but not those to Met-enk or Met-enk Arg6, also stained numerous endocrine-like cells of the antral mucosa. These were identified as gastrin cells by elution and re-staining experiments with C-terminal gastrin antisera. The rat and mouse gastrin cells might conceivably express the enkephalin gene, but fail to process it to yield Met-enkephalin; alternatively, the Met-enk Arg6Phe7-like immunoreactivity in gastrin cells could be due to a cross-reacting peptide that does not belong to the opioid series.

Animals↗

VIP-, substance P-, gastrin/CCK-, bombesin-, somatostatin- and glucagon-like immunoreactivities in the gut of the rainbow trout, Salmo gairdneri.

The presence of peptides in the gastrointestinal tract of the rainbow trout, Salmo gairdneri, was investigated immunocytochemically. VIP-like immunoreactivity was demonstrated in nerves in all layers of the stomach and the intestine, whereas substance P-like immunoreactivity was localized to endocrine cells, predominantly in the mucosa of the stomach, and to nerves mainly concentrated in the myenteric plexus throughout the gut. Endocrine cells reactive to gastrin/CCK antiserum were demonstrated in the intestinal mucosa, while no immunoreactivity was found in the stomach. Bombesin-immunoreactive and somatostatin-immunoreactive cells were localized in the stomach mucosa, and cells reactive to glucagon antiserum in the intestinal mucosa. Radioimmunoassay of stomach mucosa and muscle confirmed the presence of VIP-like and substance P-like immunoreactivity in these tissues, while gastrin/CCK-like immunoreactivity was low and bombesin-like immunoreactivity was insignificant. In conclusion, molecules resembling the mammalian brain-gut peptides may be involved in the neuronal and hormonal control of gut function in fish.

Animals↗