The relaxation induced by stimulation of the inhibitory non-adrenergic, non-cholinergic (iNANC) nerve is mediated by the release of iNANC neurotransmitters such as nitric oxide (NO), vasoactive intestinal peptide (VIP) and adenosine triphosphate (ATP). The mechanisms of NO, VIP or ATP-induced relaxation have been partly determined in previous studies, but the detailed mechanism remains unknown. We tried to identify the nature of iNANC neurotransmitters in the smooth muscle of guinea pig ileum and to determine the mechanism of the inhibitory effect of nitric oxide. We measured the effect of NO-donors VIP and ATP on the intracellular $Ca^{2+}$ concentration$([Ca^{2+}]_i)$, by means of a fluorescence dye(fura 2) and tension simultaneously in the isolated guinea pig ileal smooth muscle. Following are the results obtained. 1. Sodium nitroprusside $(SNP:10^{-5}\;M)$ or S -nitro-N-acetyl-penicillamine $(SNP:10^{-5}\;M)$ decreased resting $[Ca^{2+}]_i$ I and tension of muscle. SNP or SNAP also inhibited rhythmic oscillation of $[Ca^{2+}]_i$ and tension. In 40mM $K^+$ solution or carbachol ($(CCh:10^{-6}\;M)$-induced precontracted muscle, SNP decreased muscle tension. VIP did not change $[Ca^{2+}]_i$ and tension in the resting or precontracted muscle, but ATP increased resting $[Ca^{2+}]_i$ and tension in the resting muscle. 2. 1H-[1,2,4]oxadiazol(4,3-a)quinoxalin-1-one $(ODQ:1\;{\mu}M)$, a specific inhibitor of soluble guanylate cyclase, limited the inhibitory effect of SNP 3. Glibenclamide $(10\;{\mu}M)$, a blocker of $K_{ATP}$ channel, and 4-aminopyridine (4-AP:5 mM), a blocker of delayed rectifier K channel, apamin $(0.1\;{\mu}M)$, a blocker of small conductance $K_{Ca}$ channel had no effect on the inhibitory effect of SNP. Iberiotoxin $(0.1\;{\mu}M)$, a blocker of large conductance $K_{Ca}$ channel, significantly increased the resting $[Ca^{2+}]_i$, and tension, and limited the inhibitory effect of SNP. 4. Nifedipine $(1\;{\mu}M)$ or elimination of external $Ca^{2+}$ decreased not only resting $[Ca^{2+}]_i$ and tension but also oscillation of $[Ca^{2+}]_i$ and tension. Ryanodine $(5\;{\mu}M)$ and cyclopiazonic acid $(10\;{\mu}M)$ decreased oscillation of $[Ca^{2+}]_i$ and tension. 5. SNP decreased $Ca^{2+}$ sensitivity of contractile protein. In conclusion, these results suggest that 1) NO is an inhibitory neurotransmitter in the guinea pig ileum, 2) the inhibitory effect of SNP on the $[Ca^{2+}]_i$ and tension of the muscle is due to a decrease in $[Ca^{2+}]_i$ by activation of the large conductance $K_{Ca}$ channel and a decrease in the sensitivity of contractile elements to $Ca^{2+}$ through activation of G-kinase.
In this study, we investigated whether ambroxol significantly affects secretion, production and gene expression of mucin from cultured airway epithelial cells. Confluent primary rat tracheal surface epithelial (RTSE) cells were pretreated with adenosine triphosphate (ATP) for 5 min and then treated for 30 min with ambroxol to assess the effect on mucin secretion using ELISA. Additionally, confluent NCI-H292 cells were pretreated with ambroxol for 30 min and then stimulated with EGF or PMA for 24 h. The MUC5AC mucin gene expression and mucin protein production were measured by RT-PCR and ELISA. The results were as follows: (1) ambroxol did not significantly affect ATP-induced mucin secretion from cultured RTSE cells; (2) ambroxol inhibited the production of MUC5AC mucin protein induced by EGF and PMA in NCI-H292 cells; (3) ambroxol also inhibited the expression of MUC5AC mucin gene induced by EGF and PMA in NCI-H292 cells. This result suggests that ambroxol can inhibit the production and gene expression of MUC5AC mucin, by directly acting on human airway epithelial cells.
Kim, Jae Yeun;Park, Jung Ae;Lee, Mi Hyang;Kim, Sang Ha;Jeong, Sun Young
Journal of Digital Convergence
/
v.19
no.1
/
pp.211-216
/
2021
This study was carried out to evaluate the effects of disinfection using environmental disinfectant after having assessed the extent of contamination through microbial culture testing and the Adenosine Triphosphate Bioluminescence method among the environmental management evaluation methods used for the environment in the hospital ward of patients infected by multidrug-resistant organisms. This study was conducted with the patient wards isolated due to multidrug-resistant organisms as the environmental surface. Specimens were collected from five locations including infusion pumps, IV poles, bedside cabinets, bed railings, keyboards, and blood pressure measurement cuffs. ATP and microbial culture testing were executed prior to, immediately after, and five minutes post-disinfection. According to the result contamination of the infusion pumps was statistically significantly reduced after disinfection. In addition, the bacteria before and after disinfection reduced in IV pole, bed railing, and keyboard. That is, regular environmental surface disinfection can provide safer environments to patients against infection. Therefore, it is necessary to establish guidelines including disinfection methods and intervals for environmental surfaces by evaluating the persistence of disinfectants at various institutions in the future.
Sodium hypochlorite (NaClO) is a disinfectant widely used in hospitals and food industries because of its antimicrobial activity against not only bacteria but also fungi and virus. The antibacterial activity of NaClO lies in the maintenance of a stable hypochlorous acid (HClO) concentration, which is regulated by pH of the solution. HClO can easily penetrate bacterial cell membrane due to its chemical neutrality and the antibacterial activity of NaClO is thought to depend on the concentration of HClO in solution rather than hypochlorite ions (ClO-). In this study, we investigated the antibacterial activity of NaClO according to pH adjustment by means of time kill test and assays of Reactive Oxygen Species (ROS) and adenosine triphosphate (ATP) concentration changes before and after NaClO treatment. We also investigated that the degree of cell wall destruction through field emission scanning electron microscopy (FE-SEM). Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) exposed to 5 ppm NaClO at pH 5 exhibited 99.9% mortality. ROS production at pH 5 was 48% higher than that produced at pH 7. In addition, the ATP concentration in E. coli and S. aureus exposed to pH 5 decreased by 94% and 91%, respectively. As a result of FE-SEM, it was confirmed that the cell wall was destroyed in the bacteria by exposing to pH 5 NaClO. Taken together, our results indicate that the antibacterial activity of 5 ppm NaClO can be improved simply by adjusting the pH.
The influence of (-0-Hydroxycitrate(HCA), shown to be a competitive inhibitor of adenosine 5-triphosphate(ATP) citrate lyase, on food intake and body weight, serum triglyceride and cholosterol level, in vivo rates of fatty acid and cholesterol synthesis, and fat cell number and size was investigated. 3 groups of female, 5 weeks old Sprague Dawley rats, 8 animals each, were ad libitum meal-fed or pair-fed(3 hours from 10 : 00 to 13 : 00) AIN based high glucose diet for a total period of 8 weeks. Providing normolipidemic rats orally with 400mg of HCA formula containing approximately 20mg of HCA 1 hour prior to daily feeding schedule significantly depressed in vivo hepatic rates of fatty acid and cholesterol synthesis in the liver and adipose tissue. Serum triglyceride and cholesterol levels were significantly reduced by HCA. At the end of treatment period, the rats administered with HCA resulted in a significantly reduction in body weight gain. The reduction in weights was attributable to a significant decrease in fat cell size with a smaller extent, but not significant, reduction in fat cell number. Rats receiving HCA demonstrated less food intake than the controls ; however, this decreased caloric intake was not fully responsible for the HCA induced depression of hepatic and adipocytic lipogenesis, since experiment using pair-fed cojntrol rats showed, less magnitude but similar results. Both a anorectic and an antilipogenic properties of HCA seem to be responsible for this weight reduction activity of HCA. The outcome of this study suggest that metabolic regulation may be a feasible approach to the control of obesity and hyperlipidemia.
Several strains of spore-forming lacticacid bacteria were isolated from natural sources such as soils, cereals, and foods. The general morphological and physiological characteristics of the strain 6-4 were investigated nad compared with some other industrial strains. The effects of fructose-1,6-diphoshpate (FDP), adenosine triphosphate (ATP), and pH on the lactate dehydrogenase(LDH) activity of the strain were studied, and the changes in LDH activity and spore formation under various cultural conditions were researched. The results were as follows. 1. This strain was identified to Bacillus coagulans Hammer and distributed widely in natural sources. 2. The strain strongly converted various fermentation substrates in to L(+)-lacticacid in anaerobic conditioins, and many spores that were of great advantages to the industrial application were formed easily in the aerobic condition. 3. The LDH activity of this strain was activated by FDP and inhibited by ATP. The optimal pH for the enzyme activity was 6.0-6.5. 4. In the anaerobic culture condifion, the large amount of glucose added in the medium increased the LDH activity, but the cells were not committed to sporulate. 5. When none or a very small amount of glucose (less than 0.5%) was added to culture medium in the aerobic condition, the LDH activity was decreased and many spore were produced with final pH higher than 8.5. 6. The additioin of large amount of glucose (more than 2.0%) in aerobic culture increased the LDH activity and inhibited strongly the spore formation with final pH lower than 6.0.
Objectives: Tanshinone I is a bioactive constituent in Salvia miltiorrhiza. At present, the anti-obesity effect and mechanism of tanshinone I are not fully understood. Here we investigated the effect of tanshinone I on lipid accumulation in 3T3-L1 preadipocytes and zebrafish. Methods: Lipid accumulation and triglyceride (TG) content in 3T3-L1 cells were determined by Oil Red O staining and AdipoRed assay, respectively. The expression and phosphorylation levels of adipogenic/lipogenic proteins in 3T3-L1 cells were evaluated by Western blotting. The messenger RNA (mRNA) expression levels of adipogenic/lipogenic markers and leptin in 3T3-L1 cells were measured by reverse transcription polymerase chain reaction (RT-PCR). Lipid accumulation in zebrafish was assessed by LipidGreen2 staining. Results: Tanshinone I at 5 μM largely blocked lipid accumulation and reduced TG content in differentiating 3T3-L1 cells. Furthermore, tanshinone I decreased the expression of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), and perilipin A but also the phosphorylation of signal transducer and activator of transcription-3 (STAT-3) in differentiating 3T3-L1 cells. In addition, tanshinone I increased the phosphorylation of adenosine 3',5'-cyclic monophosphate (cAMP)-activated protein kinase (AMPK) while decreased the intracellular adenosine triphosphate (ATP) content with no change in the phosphorylation and expression of liver kinase-B1 in differentiating 3T3-L1 cells. Importantly, tanshinone I also reduced the extent of lipid deposit formation in developing zebrafish. Conclusions: These findings demonstrate that tanshinone I has strong anti-adipogenic effects on 3T3-L1 cells and reduces adiposity in zebrafish, and these anti-adipogenic effect in 3T3-L1 cells are mediated through control of C/EBP-α, PPAR-γ, STAT-3, FAS, ACC, perilipin A, and AMPK.
This study was aimed at investigation of the stimulatory innervations on the rat urinary bladder. Detrusor muscle strips of 15 mm long were suspended in isolated muscle chambers containing 1 ml of PSS maintained at $37^{\circ}C$ and aerated with 95% $O_2/5%CO_2$. Isometric myography was perfomed, and the results were as followings : Muscle strips showed "on-contraction" by electric field stimulation (EFS) frequency-dependently. The EFS-induced contraction was not affected by hexamethonium, a ganglion blocker, but abolished, by tetrodotoxin, a nerve conduction blocker. Physostigmine, a cholinesterase inhibitor enhanced the EFS-induced contraction which was inhibited by hemicholinium, an inhibitor of choline uptake at the cholinergic nerve ending. Such an EFS-induced contraction was antagonized by atropine only partially, and the atropine-resistant portion was completely abolished by the desensitization of purinergic receptors by prolonged incubatin of the strips in the presence of high concentratin of ATP. Bethanechol, a cholinergic agonist, elicited concentration-dependent contraction. Adenosine triphosphate (ATP), a purinergic agonist, induced a weak but concentration-dependent contraction of short duration. Bethanechol-induced contraction was not affected by ATP-desensitization, and ATP-induced contraction was not affected by tetrodotoxin. These results suggest that there are at least two main stimulatory components of innervations in the detrusor muscle, cholinergic muscarinic and purinergic ; and those receptors are independent each other.
Background: The replicative senescence of human dermal fibroblasts (HDFs) is accompanied by growth arrest. In our previous study, the treatment of senescent HDFs with Rg3(S) lowered the intrinsic reactive oxygen species (ROS) levels and reversed cellular senescence by inducing peroxiredoxin-3, an antioxidant enzyme. However, the signaling pathways involved in Rg3(S)-induced senescence reversal in HDFs and the relatedness of the stereoisomer Rg3(R) in corresponding signaling pathways are not known yet. Methods: We performed senescence-associated β-galactosidase and cell cycle assays in Rg3(S)-treated senescent HDFs. The levels of ROS, adenosine triphosphate (ATP), and cyclic adenosine monophosphate (cAMP) as well as the mitochondrial DNA copy number, nicotinamide adenine dinucleotide (NAD)+/1,4-dihydronicotinamide adenine dinucleotide (NADH) ratio, and NAD-dependent sirtuins expression were measured and compared among young, old, and Rg3(S)-pretreated old HDFs. Major signaling pathways of phosphatidylinositol 3-kinase/Akt, 5' adenosine monophosphate-activated protein kinase (AMPK), and sirtuin 1/3, including cell cycle regulatory proteins, were examined by immunoblot analysis. Results: Ginsenoside Rg3(S) reversed the replicative senescence of HDFs by restoring the ATP level and NAD+/NADH ratio in downregulated senescent HDFs. Rg3(S) recovered directly the cellular levels of ROS and the NAD+/NADH ratio in young HDFs inactivated by rotenone. Rg3(S) mainly downregulated phosphatidylinositol 3-kinase/Akt through the inhibition of mTOR by cell cycle regulators like p53/p21 in senescent HDFs, whereas Rg3(R) did not alter the corresponding signaling pathways. Rg3(S)-activated sirtuin 3/PGC1α to stimulate mitochondrial biogenesis. Conclusion: Cellular molecular analysis suggests that Rg3(S) specifically reverses the replicative senescence of HDFs by modulating Akt-mTOR-sirtuin signaling to promote the biogenesis of mitochondria.
Objectives: This study aimed to assess the improvement of teachers' sanitation performance and food distribution environment at Centers for children's food service management after a sanitation education program. Methods: The subjects were 119 teachers working at child care centers registered in the Daejeon Center for Children's Foodservice Management in Dongu and Jung-gu, Daejeon. The sanitation education was provided three times from March to August in 2014, and the survey questionnaires were distributed before and after the education. The sanitation status of food service environment of the centers was examined by ATP(adenosine-5'-triphosphate) bioluminescence. Results: After the sanitation education, the teachers showed higher levels of sanitation knowledge. Also, sanitation performance and recognition level of the importance of it significantly improved after the education program. The mean variation scores of importance and performance after the education were 0.14 and 0.23. According to the Importance-Performance Analysis (IPA) results of 26 sanitation attributes about the food service environment, the selection attributes with relatively low performance and importance were mostly distributed in the children's and distributer's personal hygiene management area. However, all attributes except using a personal water bottle and cup were moved to high performance and importance level. Also, the ATP examination results showed that the sanitation status of the food service environment was improved after the education program. Conclusions: The sanitation education program was effective in improving the recognition of the importance of sanitation and its performance with regard to food service management.
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