Kim, Eok Nyun;Park, Chang Hoon;Woo, Mi Na;Yoon, Ji Young;Park, Bong Soo;Kim, Yong Ho;Kim, Cheul Hong
Journal of The Korean Dental Society of Anesthesiology
/
v.14
no.2
/
pp.101-106
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2014
Background: Remifentanil, an ultra-short-acting mu-opioid receptor agonist, is unique from other opioids because of its esterase-based metabolism, minimal accumulation, and very rapid onset and offset of clinical action. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. However, the effects of remifentanil on human keratinocyte and autophagy have yet to be fully elucidated during hypoxia-reoxygenation. Here we investigated whether remifentanil confers protective effect against hypoxia-reoxygenation in human keratinocyte and, if so, whether autophagy mediates this effect. Methods: The human keratinocytes were cultured under 1% oxygen tension. The cells were gassed with 94% $N_2$, and 5% $CO_2$ and incubated for 24 h at $37^{\circ}C$. To determine whether the administration of affects human keratinocytes hypoxia-reoxygenation injury, cells were then exposed to various concentrations of remifentanil (0.01, 0.1, 0.5 and 1 ng/ml) for 2 h. After remifentanil treatment, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at $37^{\circ}C$. Control group did not receive remifentanil treatment. Normoxia group did not receive hypoxia and remifentanil treatment for 36 h. 3-MA group was treated 3-methyladenine (3-MA) for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with MTT, showing the mitochondrial activity of living cells. Cells were stained with fluorescence and analyzed with Western blot analysis to find out any relations with activation of autophagy. Results: Prominent accumulation of autophagic specific staining MDC was observed around the nuclei in RPT group HaCaT cells. Similarly, AO staining, red fluorescent spots appeared in RPT group HaCaT cells, while the Normoxia, control and 3-MA groups showed mainly green cytoplasmic fluorescence. We here examined activation of autophagy related protein under H/R-induced cells by Western blotting analysis. Atg5, Beclin-1, LC3-II (microtubule-associated protein 1 light chain 3 form II) and p62 was elevated in RPT group cells. But they were decreased when autophagy was suppressed by 3-MA (Fig. 5). Conclusions: Although the findings of this study are limited to an in vitro interpretation, we suggest that remifentanil may have a beneficial effect in the recovery of wound from hypoxia-reoxygenation injury.
Meat production and quality traits in beef cattle are largely affected by genetic factors. Acetyl-Coenzyme A carboxylase-${\alpha}$ (ACACA) plays a key role in the regulation and metabolism of fatty acid biosynthesis in mammalian animals. The gene encoding ACACA enzyme was chosen as a candidate gene for carcass and meat traits. In this study, we investigated effects of single nucleotide polymorphisms (SNPs) in the ACACA gene on beef carcass and meat traits in Hanwoo (Korean cattle) populations. We have sequenced a fragment of intron I region of the Hanwoo ACACA gene and identified two SNPs. Genotyping of the two SNP markers (g.2344T>C and g.2447C>A) was carried out using PCR-SSCP analysis in 309 Hanwoo steers to evaluate their association with carcass and meat production traits. The g.2344C SNP marker showed a significant increasing effect on LW (p = 0.009) and CW (p = 0.017). Animals with the CC genotype had higher CW and LW compared with TT and TC genotypes (p<0.05). The g.2447A SNP marker was associated with higher MC (p = 0.019). Animals with the AA genotype had higher MC than animals with CC and CA genotypes (p<0.05). Although the degree of linkage disequilibrium (LD) was not strong between g.2344T>C and g.2447C>A in the LD analysis, four major haplotype classes were formed with two SNP information within the ACACA gene. We constructed haplotypes using the HaploView software package program and analyzed association between haplotypes and carcass traits. The haplotype of ACACA gene significantly affected the LW (p = 0.027), CW (p = 0.041) and MC (p = 0.036). The effect of h1 haplotype on LW and CW was larger than that of h3 haplotype. Animals with the h1 haplotype also had greater MC than did animals with h2 haplotype. Consequently, the ACACA gene could be useful as a DNA marker for meat production traits such as carcass yield and meat contents in Hanwoo.
1. In the course of investigation on the metabolism of cysteine by microorganisms, the authors have found that among 70 strains tested Cysteinedesulfhydrase occured most remarkably in the cells of the strain I-3-2 isolated from the soil when it was grown on a medium containing L-Cysteine. The morphological, cultural and physiological properties of this strain were investigated. From the results, the bacterium was identified as a variety of Aerobactor aerogenes. 2. The cultural conditions for the formation of Cysteinedesulfhydrase by the strain I-3-2 were also investigated and the results were as follows: 1). The optium pH of the culture medium was 7.0-7.5. 2). As a carbon source, glycerol was most effective when it was added to the basal medium at 0.1 % concentration. 3). By addition of calcium chloride at 0.2% concentration, the formation of the enzyme remarkably increased. 4). Maximal formation of the enzyme was observed at the end of logarithmic phase of cell growth, thereafter the enzyme activity was diminished rapidly.
This study was designed to observe the change of body composition and nutrient metabolites and the excretion of minerals during complete fasting for 10 days in thirty women. Average loss of body weight was 7.98kg and body fat was gradually reduced after 9 days fasting, but the loss of lean body mass was reduced after 7 days fasting. Serum glucose level was sustained at constant level, but serum levels of blood urea nitrogen, free acid and $\beta$-hydroxybutyrate were significantly increased during fasting but decreased after re-feeding. Urinary excretions of 3-methylhistidine, total creatinine, and urea were high in the beginning of fasting but gradually decreased. Serum level of parathyroid hormone was significantly reduced by fasting but regained after re-feeding. Serum level of minerals was at the constant level throughout the experimental period. The urinary excretion of minerals(Ca, K, Mg, P) was significantly increased in the early stage of fasting and then reduced from 7 days, but the excretion of Zn was continuously increased until the late stage of fasting. These results showed that amino acid fatty acid released from the breakdown of muscle protein and body fat, respectively, were utilized for energy during fasting. Body weight and BMI were reduced because of the increased muscle protein breakdown and body water excretion during early stage of fasting, but the significant body fat loss was after 9 days fasting. Therefore, it could be suggested to fast for longer than 10 days if the reduction of body fat was planned by fasting, and recommed to exercise and ingest more vitamins and minerals to replace the excreted minerals during fasting. (Korean J Nutrition 34(5) : 547~553, 2001)
The effect of selenium and vitamin E on the incidence of retained fetal membranes(RFM) was evaluated in dairy cows raised in Kyonggi-do province from July through September 1985. Sodium selenite(0.1mg/kg of body weight) and vitamin E (1,000 IU) were simultaneously injected 21 days prior to the expected calving day to Holstein cows and the incidence in the treated group was compared with that in the non-treated control group. Serum levels of selenium, vitamin E, calcium, inorganic phosphorus and magnesium were also determined in the treated and the non-treated groups to compare the status of vitamin E and other minerals during periparturient period in the cows with RFM and the normal ones. The incidence of RFM was 34.5% in cows of the non-treated group (29 heads), whereas it was significantly reduced to 9.7% in cows of the treated group (31 heads) (p<0.05). Data for serum mineral concentrations showed that the prepartal inorganic phosphorus concentration was significantly lower in the RFM than in the not-retained group(p<0.01). As a result, the prepartal Ca/P ratio was significantly higher in the RFM group(p<0.01). It appears that a single injection of 0.1mg of sodium selenite per kg body weight and 1,000 IU of vitamin E 21 days prior to the expected calving day is an effective prophylactic for prevention of RFM, and that RFM may be related to imbalances in calcium and phosphorus metabolism.
This study was designed to find out the nutritional defect of general Korean diet and the effective way of nutritional supplementation. Seventy weanling Sprague-Dawley male rats weighing 51.8$\pm$0.9g were blocked into ten groups and fed ten different diets ad libitum for eight weeks: Standard groups(st gp) was given 72% sugar-20% casein diet: Cereal-vegetable group(c-v gp) was fed cereal-vegetable diet(c-v diet) composed of rice, barley, soybean, spinach and cabbage: the other eight groups were fed c-v diets supplemented with casein, vitamin $B_{2}$, calcium, vitamin A, vitamin B2 and A, vitamin A and calcium, vitamin $B_{2}$ and calcium, vitamin A and $B_{2}$and calcium, respectively, on the basis of each nutrients content of standard diet. The results were as follows: 1. Food intakes and body weight gains in all the experimental groups were significantly lower than st gp. Among experimental groups, casein gp and vit B2 gp tended to gain more body weights than c-v gp. 2. Through all the experimental period, F.E.R., pp.E.R., and NDPcal% did nod show significant differences among all the experimental groups and st gp. 3. The weights of liver, kidney, and gastrocnemius were significantly lower in all the experimental groups as compared with st gp. But brain and sex organ weights did not show differences among all the groups. 4. Nitrogen contents of total carcass, liver, and gastrocnemius in all the experimental groups tended to be increased as compared with st gp, and among experimental groups, they tended to be increased by casein supplementation and decreased by ca supplementation. 5. Apparent nitrogen digestibility, urinary and fecal nitrogen excretion, the amount of nitrogen retained, and N.P.U. did not show any significant differences among all the groups. 6. Serum total protein concentrations did not show any significant differences among all the groups.
Cellular redox state is known to be perturbed during ischemia and that $Ca^{2+}$ and $K^2$ channels have been shown to have functional thiol groups. In this study, the properties of thiol redox modulation of the ATP-sensitive $K^2$ ($K_{ATP}$) channel were examined in rabbit ventricular myocytes. Rabbit ventricular myocytes were isolated using a Langendorff column for coronary perfusion and collagenase. Single-channel currents were measured in excised membrane patch configuration of patch-clamp technique. The thiol oxidizing agent 5,5'-dithio-bis-(2-nitro-benzoic acid) (DTNB) inhibited the channel activity, and the inhibitory effect of DTNB was reversed by dithiothreitol (disulfide reducing agent; DTT). DTT itself did not have any effect on the channel activity. However, in the patches excised from the metabolically compromised cells, DTT increased the channel activity. DTT had no effect on the inhibitory action by ATP, showing that thiol oxidation was not involved in the blocking mechanism of ATP. There were no statistical difference in the single channel conductance for the oxidized and reduced states of the channel. Analysis of the open and closed time distributions showed that DTNB had no effect on open and closed time distributions shorter than 4 ms. On the other hand, DTNB decreased the life time of bursts and increased the interburst interval. N-ethylmaleimide (NEM), a substance that reacts with thiol groups of cystein residues in proteins, induced irreversible closure of the channel. The thiol oxidizing agents (DTNB, NEM) inhibited of the $K_{ATP}$ channel only, when added to the cytoplasmic side. The results suggested that metabolism-induced changes in the thiol redox can also modulate $K_{ATP}$ channel activity and that a modulatory site of thiol redox may be located on the cytoplasmic side of the $K_{ATP}$ channel in rabbit ventricular myocytes.
Background: Malignant arrhythmias require drug therapy. However, most of the currently available antiarrhythmic drugs have significant side effects. Ginsenoside Rg2 exhibits excellent cardioprotective effects and appears to be a promising candidate for cardiovascular drug development. So far, the oral toxicity and antiarrhythmic effects of Rg2 have not been evaluated. Methods: Acute oral toxicity of Rg2 was assessed by the Limit Test method in mice. Subchronic oral toxicity was determined by repeated dose 28-day toxicity study in rats. Antiarrhythmic activities of Rg2 were evaluated in calcium chloride-induced arrhythmic rats. Antiarrhythmic mechanism of Rg2 was investigated in arrhythmic rats and H9c2 cardiomyocytes. Results: The results of toxicity studies indicated that Rg2 exhibited no single-dose (10 g/kg) acute oral toxicity. And 28-day repeated dose treatment with Rg2 (1.75, 3.5 and 5 g/kg/d) demonstrated minimal, if any, subchronic toxicity. Serum biochemical examination showed that total cholesterol in the high-dose cohort was dramatically decreased, whereas prothrombin time was increased at Day 28, suggesting that Rg2 might regulate lipid metabolism and have a potential anticoagulant effect. Moreover, pretreatment with Rg2 showed antiarrhythmic effects on the rat model of calcium chloride induced arrhythmia, in terms of the reduced duration time, mortality, and incidence of malignant arrhythmias. The antiarrhythmic mechanism of Rg2 might be the inhibition of calcium influx through L-type calcium channels by suppressing the phosphorylation of Ca2+/calmodulin-dependent protein kinase II. Conclusion: Our findings support the development of Rg2 as a promising antiarrhythmic drug with fewer side effects for clinical use.
It was previously reported that the deterioration rate of contractile force of the isolated heart from ginseng extract treated rat was slower than that from control. Present study was carried out to investigate the mechanism of the action of ginseng on the contractile force of the papillary muscle in terms of calcium metabolism. Rats weighing 200-300g were administered orally with ginseng ethanol extract (100mg/kg/day) for more than 10 days. The isolated papillary muscles from rat hearts were suspended in bath containing Krebs-Henseleit solution. When equilibration of contractile force of papillary muscle was reached, the rates of deterioration of contractile forces of papillary muscle from ginseng component treated rats were determined by washing with Ca-free Krebs-Henseleit solution and compared with that of normal hearts. At the beginning of washing, the rate of deterioration of contractile force of the papillary muscle was slower significantly in ginseng treated rats than in control rats, suggesting that calcium may be somehow involved in sustaining the contractility in ginseng treated hearts. Anoxia of papillary muscle with nitrogen gas to muscles inhibited the contractility, but differences between control and ginseng treated groups in the rate of deterioration were not observed. Influence of diltiazem, calcium blocker, on the contractility of papillary muscle from ginseng treated and control hearts was studied. Contractility of papillary muscle from control and ginseng treated hearts was inhibited by diltiazem in dose dependent manner but the inhibition of the ginseng treated muscles was much weak. The effect was significantly different. From the results, it seemed that slowing in deterioration rate of papillary muscle from ginseng treated hearts might be related to calcium which was mobilized from plasma membrane of internal organelle by ginseng.
This study was performed to estimate the effect of alphacalcidol supplementation or nutrition education on the nutrient intakes, bone mineral density and bone markers in continuous ambulatory peritoneal dialysis (CAPD) patients. The 90 CAPD subjects were randomly assigned to 3 groups (alphacalcidol group: AG, nutrition education group: NG, and control group: CG). Alphacalcidol supplementation($0.5{\mu}g/day$) was carried out for 8 months. Nutrition counseling was performed according to the patient s individual question for the first 6 months and scheduled nutrition education with individual counseling was carried out for the last 2 months. In baseline data. there were no significant differences in age, sex, family number, education years and monthly income except the NG showed significantly less duration of CAPD (p< 0.05) compared to other two groups. After intervention all three groups showed tendency of lower intakes. NG revealed less decrease in protein, especially in animal protein calcium from Ca-P binder, dietary calcium, dietary iron and niacin. NG showed significantly more increase in dry weight (p<0.05) and AG in waist circumference (p<0.001) after intervention. The groups did not show significant differences in the changes of biochemical indices related to bone metabolism. NG revealed more increase in trochanter BMD(p < 0.05) compared to other two groups. It seems that nutrition education is more effective in preventing deterioration or improving the bone and general nutrition status.
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