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Query: EC:3.4.24.3 (
collagenase
)
18,340
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
The biosynthetic activity of the B-cells of obese hyperglycemic mice (obob) was measured by the incorporation of [3H]leucine into proteins in
collagenase
-isolated pancreatic islets. To quantitate the incorporation into proinsulin and
insulin
, an immune binding method was used. For this purpose, anti-
insulin
serum was coupled to cyanogen bromide-activated SepharoseR 4B. This turned out to be a specific and versatile technique for the measurement of newly synthesized proinsulin and
insulin
in the B-cells. The B-cells of obob mice appear to be well adapted to a high rate of hormone biosynthesis, since at 16.7 mM glucose 44% of [3H]leucine incorporated into TCA-precipitable proteins was bound to the
insulin
antibodies coupled to Sepharose 4B. The
insulin
biosynthetic rate was stimulated 9 times at 16.7 mM glucose, during a 3-h incubation, compared with the basal
insulin
biosynthesis rate.
...
PMID:Anti-insulin serum coupled to Sepharose 4B as a tool for the investigation of insulin biosynthesis in the B-cells of obese hyperglycemic mice. 110 96
Pancreatic islets of mice were isolated by the
collagenase
method. After a preincubation period of 20 min they were incubated in Krebs-Ringer-albumin buffer in the presence of glucose. Concanavalin A (2 mg/ml) inhibits the glucose-induced release of
insulin
; this same property is shown by Concanavalin A bound to nonphagocytosable beads of Sephrose at lower concentrations (0.4 mg/ml). In pancreatic islets stimulated by tolbutamide, glibenclamide or arginine the secretion of
insulin
is not inhibited in the presence of Concanavalin A. The incorporation of tritiated leucine is not influenced by Concanavalin A. Since desialized pancreatic islets react in the same way the glucose-induced secretion of
insulin
appears to be a membrane-dependent process.
...
PMID:Interactions of concanavalin A with isolated pancreatic islets. 110 13
The function of the pancreatic B-cell was studied in relation to the development of the diabetic syndrome in a new variety of the diabetic mutant mouse, which was produced at The Jackson Laboratory, Bar Harbor, Maine, U.S.A. by outcrossing of a c57bl/ksJ-db stock with C57BL/6J mice. The expression of the db-gene in the resulting strain was evaluated by measurements of the body weights and the concentrations of serum glucose and serum
insulin
at different ages of the animals. In the diabetic mice the body weights increased rapidly between 5 and 25 weeks of age to a weight twice that of the lean controls. During the same time hyperglycaemia and hyperinsulinaemia occurred, the maximal serum glucose and
insulin
values being observed between 17 and 25 weeks of age. Later on the serum glucose and serum
insulin
concentrations gradually decreased. Islets were isolated with
collagenase
from animals 5, 10 or 20 weeks old, and studied with respect to
insulin
content, glucose oxidation and the secretion and synthesis of
insulin
. The results were compared with data from control experiments with islets isolated from non-diabetic littermates. No major differences were found between islets from diabetic and control mice with regard to the glucose oxidation rate, whereas an exaggerated
insulin
response to glucose was observed in islets from 5 weeks old diabetic mice. In the 20 weeks old diabetic animals there was a significantly decreased islet
insulin
content and a considerably lowered
insulin
biosynthesis.
...
PMID:Function of the pancreatic B-cell during the development of hyperglycaemia in mice homozygous for the mutations "diabetes" (db) and "misty" (m). 110 67
The surfaces of isolated pancreatic islet cells were studied with the scanning and transmission electron microscopes. Islets were isolated from the pancreas of Wistar rats by
collagenase
treatment and were incubated either in glucose-free medium or in 300 mg% glucose for one hour. Immunoreactive
insulin
(IRI) in the media of both control and experimental preparations was assayed. Islets were then transferred to 4% glutaraldehyde, buffered with cacodylate, pH 7.4, and prepared for scanning and transmission electron microscopy. Cell masses average 200 mu in diameter. Alpha cells appear pyramidal in shape, are about 8 mu in diameter and appear in groups. Beta cells are round or oval in shape and have an average diameter of 10 mu. Glucose stimulation raised the IRI value tenfold and increased the number of blebs and other surface irregularities per unit area of beta cell surface. Comparison with transmission electron micrographs suggests that the blebs are related to the process of emiocytosis.
...
PMID:The surface structure of isolated pancreatic islet cells. 110 68
Using an in vitro rabbit pancreas system, we studied the effect of monoamine oxidase (MAO) inhibitors on flucose-stimulated
insulin
secretion. We evaluated the effect of both brief (15 min) and prolonged (60 min) exposure of pancreas segments to non-hydrazine (harmine, alpha-methyltryptamine, tranylcypromine and pargyline) and hydrazine (phenelzine, nialamide, iproniazid) type MAO inhibitors. All of the hydrazine type MAO inhibitors potentiated glucose-stimulated
insulin
secretion. Of the non-hydrazine inhibitors, only harmine and alpha-methyltryptamine potentiated glucose-stimulated
insulin
secretion. Hydrazine, although not itself an MAO inhibitor, also potentiated
insulin
secretion. Sixty minutes of exposure to tranylcypromine or alpha-methyltryptamine caused a decrease in
insulin
secretion. These MAO inhibitors are primary amines and primary amines can inhibit
insulin
secretion. The dopamine (DA) or serotonin (5-HT) content of the B-cells was increased by incubating rabbit pancreas with L-3, 4-dihydroxyphenylalanine (L-Dopa) or 5-hydroxytryptophan (5-HTP) for forty-five minutes prior to stimulation with glucose. Non-hydrazine MAO inhibitors increased dopamine inhibition of
insulin
secretion and either did not alter, or decreased serotonin inhibition of
insulin
secretion. Rabbit pancreatic islets were isolated using the
collagenase
digestion technique. The MAO activity of islet homogenates was determined using 5-HT and DA as substrates. Rabbit islet MAO has only one-tenth the specific activity against 5-HT (35 +/- 8.7 mumumoles/mg/min, M +/- SEM) that it has against DA (357 +/- 62.3 mumumoles/mg/min). This suggests that one reason that MAT inhibitors do not increase serotonin inhibition of
insulin
secretion is because MAO is not the major pathway for 5-HT inactivation in rabbit pancreatic islets. These studies suggest that MAO inhibitors alter
insulin
secretion, by both decreasing B-cell monoamine degradation and by mechanisms that do not involve MAO inhibition.
...
PMID:Monoamine oxidase inhibitors: nature of their interaction with rabbit pancreatic islets to alter insluin secretion. 110 23
Procedures were developed for preparing partially purified beta cell monolayer cultures. Neonatal rat pancreases were dissociated with a trypsin-
collagenase
solution. Beta cells were separated from denser acinar cells by centrifuging the initial suspension in a two layer discontinous Ficoll gradient (20, 25%). Resultant cultures, highly enriched in beta cells, were further purified by incubation with cystine-free medium. This caused necrosis of the majority of fibroblastoid cells within two days, while beta cells were considerably less affected. The resultant cultures contained an average of 72% beta cells compared to 10% in untreated control cultures.
Insulin
release by purified monolayers remained responsive to changes in the glucose concentration of the culture medium.
...
PMID:Pancreatic beta cell culture: preparation of purified monolayers. 111 78
Parenchymal rat liver cells were isolated by a modification of the
collagenase
method of Quistorff, Bondesen and Grunnet. The cells secreted albumin into the medium and incorporated 14C-leucine both into cell proteins and proteins secreted into the medium. Albumin production measured from the immunologically determined increment in the incubation medium was 1.7 +/- 0.2 mug albumin/min per g liver wet wt. This is about 30% of the rate of production in the perfused liver. Addition of
insulin
(10(-6)-10(-10) M) enhanced albumin production (50-17%), and incorporation of 14C-leucine both into albumin (50-8%), secreted proteins (40-9%) and cell proteins (20-8%).
Insulin
does not increase the production of albumin by depleting the cells. The effect of
insulin
on albumin production is compatible with an effect on the rate of synthesis as the specific activity of albumin is unaffected by addition of
insulin
.
...
PMID:Effect of insulin on albumin production and incorporation of 14C-leucine into proteins in isolated parenchymal liver cells from normal rats. 115 79
Hepatocytes were isolated by
collagenase
in vitro perfusion technique. Net glucose production in isolated hepatocytes obtained from fed, fasted and alloxan diabetic rats was studied. Net glucose production from alanine, pyruvate and fructose was increased by 2-5 fold in isolated hepatocytes obtained from fasted and alloxan diabetic rats. Similar increases in the incorporation of 14C-bicarbonate into glucose was also observed. Net glucose production in isolated hepatocytes was also compared to other in vitro preparations. Net glucose production was much higher (2-5 fold) in isolated hepatocytes than that reported previously for liver slices or perfused liver. Studies on glycogen and protein synthesis show a 2 fold stimulation in the incorporation of 1-14C-glucoase into glycogen and U-14C-leucine into protein by the addition of 100 muU of
insulin
to isolated hepatocytes.
...
PMID:Studies on gluconeogenesis and stimulation of glycogen and protein synthesis in isolated hepatocytes in alloxan diabetic, normal fed and fasted animals. 122 3
Amino acid transport was studied in primary cultures of parenchymal cells isolated from adult rat liver by a
collagenase
perfusion technique and maintained as a monolayer in a serum-free culture medium. Amino acid transport was assayed by measuring the uptake of the nonmetabolizable amino acid, alpha-aminoisobutyric acid. Rat liver parenchymal cells transported alpha-aminoisobutyric acid by an energy-dependent Na+-requiring system which displayed Michaelis-Menten kinetics. Addition of
insulin
to cultured rat liver parenchymal cells resulted in an increased influx of alpha-aminoisobutyric acid which was reflected in a higher initial rate of alpha-aminoisobutyric acid transport as well as an increased accumulation of alpha-aminoisobutyric acid at later time points. Cycloheximide effectively blocked the increase while results with actinomycin D were equivocal.
Insulin
at concentrations as low as 50 pM was effective in stimulating alpha-aminoisobutyric acid transport while the maximal response was observed at 80 nM.
...
PMID:Induction of amino acid transport in primary cultures of adult rat liver parenchymal cells by insulin. 127 Apr 36
Human hair follicles were isolated from the scalp by dispase and
collagenase
treatment and dispersed into a cell suspension by trypsin. These cells proliferated well and could be subcultured 7 to 8 times. The medium used was MCDB 153 HAA medium further supplemented with some amino acids, hydrocortisone,
insulin
, EGF, and bovine brain extract. The concentration of Ca++ was adjusted to 0.1 mM. Immunohistochemically, these cells were proved to possess keratins specific to hair forming cells.
...
PMID:In vitro keratin expression of hair cells. 128 73
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