The system most likely responsible for the accelerated metabolism of alcohol with chronic ingestion or at high blood ethanol levels, is the microsomal ethanol-oxidizing system(M EOS). While the increase in the MEOS with chronic ethanol ingestion is thought to be adaptive, it may also have serious adverse effects on the liver. The rates of the NADPH-dependent oxygen consumption by the liver microsomes from the prolonged ethanol fed rats were 2 times higher than the rates from the non-treated rats. With the alcohol ingestion, the total SH and nonprotein SH contents showed the significant decrease and at the same time, MDA in liver and GOT and GPT levels in blood showed the significant increase, which suggests the occurrence of liver damage due to the oxidative stress caused by chronic alcohol consumption. The mitochondrial aldehyde dehydrogenase(ALDH) activity was decreased by chronic ethanol ingestion, whereas the alcohol dehydrogenase activity and the cytosolic ALDH activity were not altered. These results suggest that the induction of cytochrome P450 by the chronic alcohol ingestion increases the oxidative stress which seems to result in the altered the physiological states of the liver including the ALDH activity, which may in turn to lead to the liver disease.
Rhyu Jun Ki;Yu Bong Seon;Jeong Jae Eun;Bak Jin Yeong;Son In Hwan;Lee Ju Seok;Jeon Byeong Hun;Mun Byung Soon
Journal of Physiology & Pathology in Korean Medicine
/
v.18
no.3
/
pp.900-907
/
2004
The antiproliferative effect of the water extract of the branch and root bark of Ulmi Pumilae Cortex(WEUPC) was investigated on the p53-negative human leukemia cell line (HL-60). A dose- and time-dependent inhibition of cell growth was observed; this effect appears to be due to induction of apoptosis. Involvement of oxidative stress is indicated by a dose-dependent increase in intracellular reactive oxygen species levels. In addition. anti-apoptic effect was observed in the cells simultaneously treated with WEUPC and the anti-oxidant N-acetylcysteine. WEUPC did not affect the anti-apoptotic Bcl-2 and the pro-apoptotic Bax, whereas p21/sup WAF1/CIPl/ was enhanced in a dose- and time-dependent fashion; this effect was partially inhibited by N-acetylcysteine. The increase in p21/sup WAF1/CIPl/ was accompanied by a parallel accumulation of cells in the G1 phase of the cycle. These results suggest that the p53-independent induction of p21/sup WAF1/CIP/ and the induction of apoptosis may mediate the anti proliferative effect of WEUPC at least in this study; on the basis of this observation, WEUPC could be proposed as an useful adjunct to the treatment of p53-deficient tumors, which are often refractory to standard chemotherapy.
We investigated the antioxidative effects of solvent extracts of doenjang fermented using Bacillus subtilis DJI (DJI doenjang) in vitro. The solvents used for extraction were ethanol, n-hexane, and water. The antioxidative activities of DJI doenjang solvent extracts were measured by estimation of peroxide value, the presence of linoleic acid level, and nitrite scavenging activity, the Rancimat test, and 1,1-diphenyl-2-picryl hydrazyl (DPPH) free-radical generation, in comparison with the commercial antioxidant butylated hydroxytoluene (BHT). The peroxide value of an ethanol extract was lower than those obtained using n-hexane and water extracts. Furthermore, the peroxide value of the ethanol extract was similar to that obtained after BHT treatment. The nitrite scavenging activity was 23.36% after addition of 600 ppm DJI doenjang ethanol extract, and the DPPH free-radical scavenging activity was 19.06% under same condition, which shows that DJI doenjang ethanol extract exhibited lower antioxidative capacities than did BHT. In the Rancimat test, the ethanol extract (11.20 min induction time), n-hexane extract (7.58 min induction time), and water extract (8.26 min induction time) after treatment with 600 ppm DJI doenjang extracts demonstrated longer induction periods than did BHT (6.94 min). These results indicate that DJI doenjang has potential anti-oxidative activity.
The Saccharomyces0 cerevisiae KNU5377 strain, which was isolated from spoilage in nature, has the ability to convert biomass to alcohol at high temperatures and it can resist against various stresses [18, 19]. In order to understand the defense mechanisms of the KNU5377 strain under menadione (MD) as oxidative stress, we used several techniques for study: peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) followed by two-dimensional (2D) gel electrophoresis, liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), and surface-enhanced laser desorption ionization-time of flight (SELDI-TOF) technology. Among the 35 proteins identified by MALDI-TOF MS, 19 proteins including Sod1p, Sod2p, Tsa1p, and Ahp1p were induced under stress condition, while 16 proteins were augmented under normal condition. In particular, five proteins, Sod1p, Sod2p, Ahp1p, Rib3p, Yaf9p, and Mnt1p, were induced in only stressed cells. By LC-ESI-MS/MS analysis, 37 proteins were identified in normal cells and 49 proteins were confirmed in the stressed cells. Among the identified proteins, 32 proteins were found in both cells. Five proteins including Yel047cp and Met6p were only upregulated in the normal cells, whereas 17 proteins including Abp1P and Sam1p were elevated in the stressed cells. It was interesting that highly hypothetical proteins such as Ynl281wp, Ygr279cp, Ypl273wp, Ykl133cp, and Ykr074wp were only expressed in the stressed cells. SELDI-TOF analysis using the SAX2 and WCX2 chips showed that highly multiple-specific protein patterns were reproducibly detected in ranges from 2.9 to 27.0 kDa both under normal and stress conditions. Therefore, induction of antioxidant proteins, hypothetical proteins, and low molecular weight proteins were revealed by different proteomic techniques. These results suggest that comparative analyses using proteomics might contribute to elucidate the defense mechanisms of KNU5377 under MD stress.
In order to elucidate the role of platelet activating factor (PAF) in the acute lung injury induced by endotoxin (ETX), activities of phospholipase A2, lyso PAF acetyltransferase and oxidative stress by neutrophilic respiratory burst were probed in the present study. To induce acute lung injury, $100\;{\mu}g$ of E.coli ETX (type 0127; B8) was instilled directly into the tracheae of Sprague-Dawley rats. Five hours after the ETX instillation, induction of acute lung injury was confirmed by lung leak index and protein contents in the bronchoalveolar lavage (BAL) fluid. At the same time, lung phospholipase A2 (PLA2) activity and expression of group I and II secretory type PLA2 were examined. In these acutely injured rats, ketotifen fumarate, known as lyso PAF acetyltransferase inhibitor and mepacrine were administered to examine the role of PAF in the pathogenesis of the acute lung injury. To know the effect of the ETX in the synthesis of the PAF in the lungs, lyso PAF acetyltransferase activity and PAF content in the lungs were measured after treatments of ETX, ketotifen fumarate and mepacrine. In addition, the role of neutrophils causing the oxidative stress after ETX was examined by measuring lung myeloperoxidase (MPO) and enumerating neutrophils in the BAL fluid. To confirm the oxidative stress in the lungs, pulmonary contents of malondialdehyde (MDA) were measured. After instillation of the ETX in the lungs, lung leak index increased dramatically (p<0.001), whereas mepacrine and ketotifen decreased the lung leak index significantly (p<0.001). Lung PLA2 activity also increased (p<0.001) after ETX treatment compared with control, which was reversed by mepacrine and ketotifen (p<0.001). In the examination of expression of group I and II secretory PLA2, mRNA synthesis of the group II PLA2 was enhanced by ETX treatment, whereas ketotifen and WEB 2086, the PAF receptor antagonist, decreased the expression. The activity of the lysoPAF acetyltransferase increased (p<0.001) after treatment of ETX, which implies the increased synthesis of PAF by the remodelling of lysoPAF in the lungs. Consequently, the contents of the PAF in the lungs were increased by ETX compared with control (p<0.001), while mepacrine (p<0.001) and ketotifen (p<0.01) decreased the synthesis of the PAF in the lungs of ETX treated rats. The infiltration of the neutrophils was confirmed by measuring and enumerating lung MPO and the neutrophils in the BAL fluid respectively. Compared with control, ETX increased lung MPO and number of neutrophils in BAL significantly (p<0.001) whereas mepacrine and ketotifen decrerased number of neutrophils (p<0.001) and MPO (p<0.05, p<0.001, respectively). The lung MDA contents were also increased (p<0.001) by ETX treatment, but treatment with mepacrine (p<0.001) and ketotifen (p<0.01) decreased the lung MDA contents. Collectively, we conclude that ETX increases PLA2 activity, and that the subsequently increased production of PAF was ensued by the remodelling of the lyso PAF resulting in tissue injury by means of oxidative stress in the lungs.
Objective: p66Shc, a 66 kDa protein isoform encoded by the proto-oncogene SHC, is an essential intracellular redox homeostasis regulatory enzyme that is involved in the regulation of cellular oxidative stress, apoptosis induction and the occurrence of multiple age-related diseases. This study investigated the expression profile and functional characteristics of p66Shc during preimplantation embryo development in sheep. Methods: The expression pattern of p66Shc during preimplantation embryo development in sheep at the mRNA and protein levels were studied by quantitative real-time polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The effect of p66Shc knockdown on the developmental potential were evaluated by cleavage rate, morula rate and blastocyst rate. The effect of p66Shc deficiency on reactive oxygen species (ROS) production, DNA oxidative damage and the expression of antioxidant enzymes (e.g., catalase and manganese superoxide dismutase [MnSOD]) were also investigated by immunofluorescence staining. Results: Our results showed that p66Shc mRNA and protein were expressed in all stages of sheep early embryos and that p66Shc mRNA was significantly downregulated in the 4-to 8-cell stage (p<0.05) and significantly upregulated in the morula and blastocyst stages after embryonic genome activation (EGA) (p<0.05). Immunofluorescence staining showed that the p66Shc protein was mainly located in the peripheral region of the blastomere cytoplasm at different stages of preimplantation embryonic development. Notably, serine (Ser36)-phosphorylated p66Shc localized only in the cytoplasm during the 2- to 8-cell stage prior to EGA, while phosphorylated (Ser36) p66Shc localized not only in the cytoplasm but also predominantly in the nucleus after EGA. RNAi-mediated silencing of p66Shc via microinjection of p66Shc siRNA into sheep zygotes resulted in significant decreases in p66Shc mRNA and protein levels (p<0.05). Knockdown of p66Shc resulted in significant declines in the levels of intracellular ROS (p<0.05) and the DNA damage marker 8-hydroxy2'-deoxyguanosine (p<0.05), markedly increased MnSOD levels (p<0.05) and resulted in a tendency to develop to the morula stage. Conclusion: These results indicate that p66Shc is involved in the metabolic regulation of ROS production and DNA oxidative damage during sheep early embryonic development.
Seasoned laver Pyropia spp. is one of the most well-known Korean traditional seafoods, and is becoming more popular worldwide. Various mixed oils are used in the preparation of seasoned laver; however, there is no information available regarding the effects of the blending ratio of oils on the quality of seasoned laver. In this study, the effects of the blending ratio of corn, sesame, and perilla oils on the oxidation and sensory quality of seasoned laver were monitored and optimized using a response surface methodology. An increase in the proportion of corn and sesame oils resulted in an excellent oxidation induction time, whereas a high ratio of perilla oil reduced the thermal oxidative stability of the mixed oil. In the sensory test, the seasoned laver with the highest proportion of sesame oil was preferred. The optimal blending ratio (v/v) of corn, sesame, and perilla oils for both oxidation induction time ($Y_1$) and sensory score ($Y_2$) was 92.3, 6.0, and 1.7%. Under optimal conditions, the experimental values of $Y_1$ and $Y_2$ were $4.41{\pm}0.3h$ and $5.58{\pm}0.8$points, and were similar to the predicted values (4.34 h and 5.13 points). Our results for the monitoring and optimization of the blending ratio provide useful information for seasoned laver processing companies.
Kim, Jae-Wook;Hong, Ki-Ju;Chung, Byoung-Sang;Hur, Jong-Wha
Korean Journal of Food Science and Technology
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v.29
no.2
/
pp.261-265
/
1997
To know the availability of double fractionated palm olein (DFPL) for vegetable oil in commercial mayonnaise preparation, oxidative stability and stability to cold test of DFPL, soybean oil and blended oils (blended soybean oil with DFPL) were tested. Mayonnaises with these oils were prepared and then emulsion stability at low temperature $(-5^{\circ}C)$ were compared. The oxidative stability of vegetable oil by Rancimat test showed that induction time of DFPL (26.9 hr) was longer when compared with soybean oil (13.4 hr), and became longer with increase of DFPL ratio in the blended oil. Emulsion stability of mayonnaises at low temperature $(-5^{\circ}C)$ was decreased with the increase of DFPL ratio in the blended oil. But, mayonnaise with blended oil of below 20% DFPL was comparable to that with soybean oil only. Among quality characteristics of mayonnaises with soybean oil and blended oil (soybean oil 85% plus DFPL 15%) the latter showed stronger oxidative stability and less flavor reversion during high temperature treatment. This result suggested that the possibility of DFPL to substitute for vegetable oil in the preparation of commercial mayonnaise.
When NIH3T3 cells were exposed to mild heat and recovered at $37^{\circ}C$ for various time intervals, they were thermotolerant and resistant to subsequent stresses including heat, oxidative stresses, and antitumor drug methotrexate which are apoptotic inducers. The induction kinetics of apoptosis by stresses were determined by DNA fragmentation and protein synthesis using $[35^S]$methionine pulse labeling. We investigated the hypothesis that thermotolerant cells were resistant to apoptotic cell death compared to control cells when both cells were exposed to various stresses inducing apoptosis. The cellular changes in thermotolerant cells were examined to determine which components are involved in this resistance. At first, the degree of resistance correlates with the extent of heat shock protein synthesis which were varied depending on the heating times at $45^{\circ}C$ and recovery times at $37^{\circ}C$after heat shock. Secondly, membrane permeability change was observed in thermotolerant cells. When cells prelabeled with $[^{3}H]$thymidine were exposed to various amounts of heat and recovered at $37^{\circ}C$ for 1/2 to 24 h, the permeability of cytosolic $[^{3}H]$thymidine in thermotolerant cells was 4 fold higher than that in control cells. Thirdly, the protein synthesis rates in thermotolerant and control cells were measured after exposing the cells to the same extent of stress. It turned out that thermotolerant cells were less damaged to same amount of stress than control cells, although the recovery rates are very similar to each other. These results demonstrate that an increase of heat shock proteins and membrane changes in thermotolerant cells may protect the cells from the stresses and increase the resistance to apoptotic cell death, even though the exact mechanism should be further studied.
The chemical characteristics of seed oils of Asian ginseng (Panax ginseng C.A. Meyer) at different ages grown in Korea (3, 4 and 5-year old) and China (5-year old), and American ginseng (Panax quinquefoliu L., 5-year old) grown in China were compared. Total fatty acid composition showed a significantly higher oleic acid content in American (87.50%) than in Korean (68.02~69.14%) and Chinese ginseng seed oils (61.19%). At the sn-2 position, the highest oleic acid (81.09%) and lowest linoleic acid (15.77%) were found in American ginseng seed oil. The main triacylglycerol species in ginseng seed oils were triolein (OOO) and 1,2-dioleoyl-3-linoleoyl-glycerol (LOO)/1,3-dioleoyl-2-linoleoyl-glycerol (OLO). In addition, the seed oils possessed an ideal oxidative stability showing 16.55~23.12 hr of induction time by Rancimat test. The results revealed that ginseng seed oil could be developed as a new healthy edible oil, and that the oil's chemical characteristics were strongly associated with the ginseng species and habitats.
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