Disturbance of mouse pancreatic beta-cell function following echo 4 virus infection.
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Biomedical subjects
Publications and source records attributed to K W Taylor.
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The effects of a diabetogenic strain of encephalomyocarditis (EMC) virus on total protein and insulin biosynthesis in mouse islets of Langerhans have been studied in tissue culture. In dispersed mouse islets, the rates of protein biosynthesis were assessed by measuring the incorporation of [3H]leucine into proteins. In infected dispersed islets incubated in 20 mM-glucose, both insulin and total protein biosynthesis were decreased at 6 h; only insulin biosynthesis was significantly decreased at 3 h. In whole islets, EMC virus brought about a decrease in glucose-stimulated protein and insulin biosynthesis as early as 2 h after infection without concomitant effects on insulin release. This inhibition of protein biosynthesis was still apparent at 20 h post-infection, at which time insulin release was found to be markedly elevated, and the islet insulin content was moderately decreased. At 44 h post-infection, glucose-induced insulin biosynthesis was preferentially inhibited. Infected islets at this later time point also displayed elevated levels of insulin release, and a marked loss of islet insulin content. When insulin mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA levels were assessed by dot-blot hybridization using appropriate cDNA probes, levels of insulin mRNA were shown to decrease steadily during the first 20 h of infection, in contrast with the levels of GAPDH mRNA. At 44 h post-infection, both types of mRNA were markedly decreased. It is suggested that there is an initial early 'shut-off' of protein synthesis without other detectable changes in islet function. This is followed by a phase where both insulin mRNA levels and insulin synthesis are dramatically decreased.
Infections with Coxsackie viruses (especially Coxsackie B4) are thought to be involved in the pathogenesis of diabetes. Many interdependent variables determine the outcome of an infection with a Coxsackie virus, one of them being the tropism of the virus for a specific tissue. The extent to which Beta cell tropic variants of Coxsackie B4 virus occur naturally was assessed. Human isolates of this virus were tested in an in vitro system in which elevated insulin release from infected islets incubated at a non-stimulatory (2 mmol/l) glucose concentration appears to be related to viral attack. Using this technique, 8/24 isolates tested, impaired secretory function in mouse islets. Some strains of Coxsackie B4 virus, therefore, will directly infect mouse islets in vitro leading to changes in islet cell function. In conclusion, these findings confirm that variants of Coxsackie B4 virus with the potential to damage Beta cells occur quite frequently in the natural population. In certain circumstances the damage they inflict on Beta cells may cause destruction of these cells, or precipitate overt diabetes.
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A LiF thermoluminescent dosimetry (TLD) system, calibrated in the tissue of interest with the beam used for experimentation, was employed to investigate dosages (muGy) to the thyroid region of an anthropomorphic phantom resultant from two dental complete-mouth radiographic procedures. Both techniques were compared in terms of dosages associated with combinations of lead apron and thyroid collar shielding while using a 70-kVp or 90-kVp x-ray beam for a 20-film complete-mouth series. Lead shielding significantly decreased the dose to the thyroid using both techniques (p less than 0.05). The use of the 90-kVp beam resulted in a significant reduction in the thyroid absorbed dose when using the bisecting angle technique (p less than 0.05) but caused a significant increase in the thyroid absorbed dose when the paralleling technique was used (p less than 0.05). The implementation of higher kilovoltage techniques in dental offices must therefore be dependent on the radiographic technique employed.
When rat pancreatic islets were incubated with 10(-8) M arginine vasopressin in the presence of 15 mM glucose there was a pronounced inhibition of insulin release in comparison with controls. This inhibitory effect appeared to be specific for vasopressin since it was antagonised by vasopressin antibody. Moreover, pertussis toxin (100 ng/ml) reversed the inhibition of insulin release due to vasopressin, indicating the possible involvement of a guanyl-nucleotide regulatory protein in the inhibitory effect. Nevertheless, 10(-8) M vasopressin increased islet concentrations of cyclic AMP even under conditions where insulin release was decreased.
Mouse pancreatic islets cultured in vitro were infected with a tissue culture-adapted or a mouse pancreas-adapted strain of Coxsackie B4 (CB4) virus. The effects of the viruses on the islets were assessed by examination of their biochemical functions. It was found that the mouse pancreas-adapted strain of CB4 induced a 'leakage' of insulin from islets incubated at a basal (2 mmol l-1) glucose concentration, both at two and four days following infection. However, at a stimulatory concentration of glucose (20 mmol l-1) the rate of insulin secretion appeared to be normal in these islets. At two days the rate of total protein synthesis in islets infected with mouse pancreas-adapted CB4, incubated at high glucose concentration, was reduced; at four days the degree of inhibition was more severe, the rate at basal glucose concentration falling to half that of the control islets and at the stimulatory glucose concentration to a quarter of the control islets. (Pro)insulin biosynthesis was also inhibited, the rate being reduced to less than half the mean control value in islets infected with mouse pancreas-adapted CB4 virus at 20 mmol l-1 glucose at two days; at four days the rate was greatly reduced at both 2 and 20 mmol l-1 glucose. It is concluded from this study that only certain strains of CB4 virus can infect mouse pancreatic islets in vitro and that infection with strains of virus tropic for the islets leads to an impairment of metabolic functions of the B-cells, and is not necessarily lytic.
The direct effects of alpha- and beta-interferons on isolated mouse pancreatic islets were investigated in vitro and found to be similar. After 7 h incubation with interferon concentrations above 350 units/ml, glucose-stimulated (pro)insulin biosynthesis was significantly inhibited, with only a slight inhibition of total protein biosynthesis. Inhibition could be abolished in the additional presence of an anti-interferon antibody. Interferon did not affect insulin release, total insulin content, or glucose oxidation of the islets. The stimulation of (pro)insulin biosynthesis by adenosine, D-glyceraldehyde, mannose, N-acetylglucosamine and leucine was also inhibited by interferon, with no effect on insulin release. At concentrations of dsRNA (double-stranded RNA) said to induce interferon (1-100 micrograms/ml), glucose-stimulated (pro)insulin biosynthesis was inhibited without significantly affecting insulin release. The dsRNA may itself inhibit stimulated (pro)insulin biosynthesis or may function indirectly by the induction of interferon.
Glucose-stimulated (pro)insulin biosynthesis is markedly inhibited is mouse pancreatic islets incubated with micromolar concentrations of 2-5A 'core', (A2'p5'A2'p5'A). Total protein synthesis was also reduced, but to a lesser extent, while insulin release and total insulin content of the islets was untouched by 2-5A 'core'. Evidence is given for the presence of a 2-5A-dependent endoribonuclease which mediates these effects.
Isolated mouse pancreatic islets were infected in vitro with two strains of Coxsackie B4 virus--a tissue culture-adapted strain and a mouse pancreas-adapted strain. Within 48 h of infection changes had occurred in the biochemical activities of islets infected with the mouse pancreas-adapted strain of virus. Basal insulin release was increased two-fold in these islets, while glucose-induced insulin secretion remained unchanged. Insulin biosynthesis was greatly reduced at a stimulatory concentration of glucose (20 mM), thus leading to a reduced insulin content in these islets. These effects are of importance because they demonstrate that certain strains of Coxsackie B4 virus, like encephalomyocarditis virus, may selectively alter beta-cell function in vitro.
To examine the influence of stimulus significance on more sustained as well as transient aspects of electrodermal, cardiac (HR), and eyeblink response, a 21-sec tone was sounded in one ear or the other. A click occurred during many tones, and a light followed offset by 9 sec. Four groups were studied: one pressed a pedal immediately on hearing any click; another only on click during tone in a specified ear; a third also responded only to the specified ear, but withheld press until the light; a fourth listened without any response. Results confirmed the important role of stimulus significance in each system whether between- or within-subject comparisons were made. Sustained responses were seen only when a significant signal was sought, involving in each case sustained HR deceleration, slowed blink rate, and heightened electrodermal level. Transient response to click and light also appeared only when there were significant signals. Response to tone-onset gave more ambiguous results. ANOVAs of response magnitude suggested that onset of nonsignificant tones might have elicited ORs, while binomial tests indicated these were not elicited with better than random frequency anywhere but on those trials occurring more frequently at the experiment's onset. Interpretations consistent with both the significance hypothesis and with a distinction between automatic and voluntary ORs can be made only here. Motor response had no effect on electrodermal or eyeblink response, and on HR was associated only with increased acceleration 1-2 sec after pedal-press. Studies using small motor responses to establish stimulus significance are therefore not likely to be substantially biased by the response itself.
Beta-Cell function in human islets derived from a number of kidney donors was investigated by using various types of islet preparations. With fresh islets, both insulin release and biosynthesis were increased by raising glucose concentrations, although the response was a variable one. In fresh islets, the effects of 5 mmol of glucose/l on release were potentiated by 10 mmol of D-3-hydroxybutyrate/l. Insulin release at 20 mmol of glucose/l was inhibited by adrenaline (0.1 mmol/l), and potentiated by theophylline (10 mmol/l) in the presence of 5 mmol of glucose/l, in islets cultured for 4 days. After culture for 8 days, islets still showed an increase in insulin release and biosynthesis in response to glucose. Pancreas slices derived from fresh human tissue also responded to increasing concentrations of glucose with a sigmoidal curve for insulin release.
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X-ray exposures to patients were measured in 30 rooms in Toronto hospitals. For a given procedure, the total exposure for a satisfactory examination differed from one room to another by as much as a factor of 30, skin exposures reaching 90 R and more for barium enemas and barium meals. The factors primarily responsible for these large differences in exposure were fluoroscopic exposure rate (0.65-12 R/min) and time 1.5-12 min), kVp and filtration (8:1), choice of screen-film combination (6:1) and attenuation of table tops and phototimers (4:1). The increase by a factor of 5 in exposure of pelvic organs, when using beams directed AP instead of PA, requires that remote-controlled GI machines be used cautiously, particularly on women of childbearing age. It is also important that these machines have tables with low attenuation. The use in some rooms of minimal fluoroscopic exposure rates and 70 mm fluorography at total exposures of less than 5 R shows that radiographic examinations can be made virtually "safe" with modern, properly adjusted equipment.
1. Rabbit islets of Langerhans were disrupted by ultrasonic methods and the sonicated preparations were used to study proinsulin biosynthesis. 2. When [3h]leucine is incubated in such preparations, incorporation takes place into proinsulin, as evidenced by characterization on polyacrylamide gels, and by the conversion of this labelled material into insulin, by using trypsin. 3. The labelled proinsulin may also be purified by antiinsulin antibody bound to Sepharose. 4. With the broken-cell preparation it was shown that incorporation of leucine is accelerated by increasing the glucose content of the medium from 2mM to 16mM. However, 16mM-galactose or -sucrose did not stimulate incorporation significantly from basal values. This effect of glucose was abolished by cycloheximide. 5. The significance of these findings in relation to the mechanism of glucose stimulation of proinsulin biosynthesis is discussed.