• Molecules and Cells
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• v.39 no.10
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• pp.768-775
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• 2016

The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.

• Toxicological Research
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• v.8 no.2
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• pp.265-272
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• 1992

Effects of repeated physical exercise on the carbon tetrachloride ($CCl_4$) hepatotoxicity were examined in adult female rats. Rats were introduced into a cylindrical rotating cage and forced to exercise for 1 hr each day, 6days/week, for 5 consecutive weeks at a speed starting from 10m/min, increased by 1m/min per day until the speed reached 27m/min. Significantly less body weight gain was observed in the exercise group suggesting that physical fitness had been induced in these animals. Eighteen hours following termination of the last exercise bout rats were treated with $CCl_4$(2 mmol/kg.ip). The $CCl_4$-induced heptotoxicity was significantly potentiated in the repeated exercise group compared to the resting sedentary animals as determined by changes in serum sorbitol dehydrogenase (SDH), glutamic oxaloacetic transaminase(GOT), glutamic pyruvic transaminase (GPT), and glucose-6-phosphatase(G-6-Pase) activities when measured 24hrs following the $CCl_4$ treatment. Hepatic drug metabolizing activity was determined in order to elucidate the underlying mechanism of potentiating action of the $CCl_4$ hepatotoxicity induced by repeated physical exercise. Repeated exercise increased the hepatic microsomal cytochrome P-450 contents and aminopyrine N-demethylase activity. The results suggest that the potentiation of $CCl_4$ hepatotoxicity by repeated exercise is associated with induction of the mixed function oxidase (MFO) enzyme system mediating the metabolism of $CCl_4$ to its active metabolite(s).

• Journal of Animal Science and Technology
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• v.54 no.3
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• pp.157-163
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• 2012

The male reproduction is precisely controlled by a number of intrinsic and extrinsic factors. These factors usually involve in expressional regulation of various molecules influencing on sperm production in the testis. A number of ways are employed to control the transcription of specific genes, including epigenetic modifications of DNA and histone molecules. DNA methylation of CpG dinucleotides is a commonly used regulatory mechanism for testicular genes associated with the fertility. Previous studies have demonstrated the infertility induced by improper DNA methylation of these genes. In the present research, we attempted to determine transcriptional expression of some of these genes in the rat testis at different postnatal ages using real-time PCR analysis. These genes include neurotrophin 3 (Ntf3), insulin-like growth factor II (Igf2), JmjC-domain-containing histone demethylase 2A 1 (Jhm2da), paired box 8 transcription factor (Pax8), small nuclear ribonucleoprotein polypeptide N (Snrpn), and 5,10-methylenetetrahydrofolate reductase (Mthfr). The expression levels of Ntf3, Igf2, and Snrpn genes were the highest at the neonatal age, followed by transient decreases at the prepubertal age. Expression of Jhm2da and Mthfr genes were continuously increased from the neonate to 1 year of age. The levels of Pax8 mRNA at the early ages were higher than those at the later ages of postnatal development. These findings suggest that expression of some fertility-associated testicular genes in the rat during postnatal period could be differentially regulated by the control of the degree of DNA methylation.

• Korean Journal of Environmental Biology
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• v.26 no.2
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• pp.109-114
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• 2008

To evaluate the extent of Sea-Nine 211 (4,5-dichloro-2-n-octyl-3(2H) isothiazolone), one of the alternating organic booster compound used in antifouling paint to replace TBT, on marine benthic bivalve, we injected Sea-Nine 211 to clam, Pseudocardium sachalinensis, and then determined some xenobiotics metabolizing enzyme activities, especially EROD (ethoxyresorufin deethylase) and MROD (methoxyresorufin demethylase), in digestive gland during 4 day-exposure period. Moreover, the results were compared with those of TBT exposed clam. CYP1A1 dependant EROD activity in both the Sea-Nine 211 and the TBTC exposure groups showed no significant differences compared to those of the solvent control group. CYP1A2 dependant MROD activity in Sea-Nine 211 exposure group was significantly induced, but no significant difference was obtained in the TBTC exposure group. These results indicate that Sea-Nine 211 demonstrated a tendency to induce MROD activity, while TBTC inhibits the activities of this enzyme.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.43 no.5
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• pp.428-436
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• 2010

Sea-urchins (Anthocidaris crassispina) are widely distributed in the East Sea of Korea. The aim of this study was to evaluate the hepatoprotective effects of sea-urchin roe on bromobenzene (BB)-induced liver damage in rats. The antioxidative and detoxifying properties of sea-urchin roe in BB-poisoned rat liver was examined by chemical analysis of serum aminotransferase (AST, ALT), glutathione S-transferase (GST), $\gamma$-glutamylcystein synthetase, glutathione reductase, epoxide hydrolase, amino-N-demethylase (AD), aniline hydrolase (AH) enzyme activity, as well as lipid peroxide and glutathione contents. Sea-urchin roe inhibited the increase of serum AST, ALT enzyme activity. Increasing lipid peroxide contents and AD and AH activities were significantly decreased in ethanol extract of sea-urchin roe. GST, $\gamma$-glutamylcystein synthetase, glutathione reductase and epoxide hydrolase enzyme activities increased in sea-urchin roe-fed group, compared with the BB-treated group. These results suggest that sea-urchin roe facilitates recovery from liver damage by enhancing antioxidative defense mechanisms and hepatic detoxication metabolism.

• Fisheries and Aquatic Sciences
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• v.20 no.9
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• pp.22.1-22.6
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• 2017

The antihyperlipidemic and antioxidant activities of dietary supplementation of sea tangle from Goseong and the alginate-free residue of sea tangle were investigated in Sprague Dawley rats treated with a high-fat diet, streptozotocin, poloxamer 407, and bromobenzene. The alginate-free residue of Goseong sea tangle induced a significant reduction in triglycerides and total cholesterol levels, as well as a significant increase in high-density lipoprotein cholesterol levels. Alginate-free Goseong sea tangle residue reduced the activities of the phase I enzymes aminopyrine N-demethylase and aniline hydroxylase, which had been increased by intraperitoneal injection of bromobenzene. Pretreatment with Goseong sea tangle residue prevented a bromobenzene-induced decrease in epoxide hydrolase activity. Bromobenzene reduced hepatic glutathione content and increased hepatic lipid peroxide levels. Pretreatment with alginate-free Goseong sea tangle residue prevented lipid peroxidation induced by bromobenzene, but pretreatment with Goseong sea tangle did not. These results suggest that Goseong sea tangle residue exerted antihyperlipidemic and antioxidant activities that were higher than those induced by alginate-containing sea tangle. Therefore, the alginate-free residue may contain physiologically unknown active components, other than alginic acid, which may potentially be used to prevent hyperlipidemic atherosclerosis.

• The Journal of Korean Medicine
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• v.16 no.2 s.30
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• pp.320-336
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• 1995

SANGNYANGHYOLTANG(SYT) is one of the most important prescription that has been used in oriental medicine for diabetes mellitus. The sudy was done in order to elucidate the anti-diabetic effect of SYT. After pretreatment of SYT(1,000mg／kg) for 6 weeks, the effect of of SYT was prevented on serum liver function test and hepatic lipid peroxide content in rats i.v. injected with streptozotocin(STZ, 50mg／kg, tail vein) 5 weeks after pretreatment of SYT. The hepatic microsomal cytochrome P-450 and aniline hydroxylase were significantly decreased, and aminopyrine N-demethylase activity was significantly increased in SYT-STZ group as compared with control group. Changes in aldehyde oxidase, xanthine oxidase, superoxide dismutase, catalase, epoxide hydrolase, UDP-glucuronyltransferase and sulfotransferase activities were not significantly different in any of the group. The cytosolic glutathione S-transferase activity was significantly decreased in SYT-STZ group as compared with control group. The selenium-independent glutathione peroxidase was significantly increased in SYT-STZ group as compared with control group, but there was no significant difference in selenium-dependent glutathione peroxidase in any of the groups. The hepatic glutathione concentration was significantly increased in SYT-STZ group as compared with control group, and ${\gamma}-glutamylcystein$ synthetase and glutathione reductase activities were not significantly different in any of the groups. The hepatic lipid peroxide content, serum aminotransferase and sorbitol dehydrogenase activities were slightly decreased in significantly in SYT-STZ groups.

• Journal of Ginseng Research
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• v.13 no.1
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• pp.37-41
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• 1989

Reactive metabolites generated by benzo(a)pyrene(BP) monooxygenase(AHH) interact with nucleophiles in DNA and cause mutation and carcinogenesis. We studied the effect of Panax ginseng C.A. Meyer, which induce epoxide hydratase(EH) activity without concomitant induction of AHH activity, on the binding of BP metabolites to DNA in uitro in Sprague Dawley rats. DNA-BP metabolite adducts can be resolved into at least five distinct peaks by elution of a Sephadex LH-20 column with a water methanol gradieNt. These peaks are arbitrarily designated A(most polar) through I(least polar). Of the 5 peaks tentatively assigned to 7,8 biol-9,10-oxide(A),7,8·oxide(B),4,5-oxide(C), and further metabolites of 9-OH-BP(D & E), peaks A, C, D, and I were reduced to 70, 85, 80, and 30% of controls, respectively, and there was no significant change in peak B. In connection with this DNA binding study, BP metabolizing enzymes including AHH, EH, demethylase(DM) activity and cyt. P-450 contents were also investigated in order to compare the BP treated control with ginseng and BP treated test groups. The results showed that the EH activity was increased by 139% over the BP control, the Cyt. P-450 content was increased by 180% over the control value, and DM and AHH activities were also increased to some degree for the BP test group, but there was no significant effect of the ginseng treatment.

• YAKHAK HOEJI
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• v.46 no.5
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• pp.358-363
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• 2002

In Oriental medicine, the prescriptions composed of several herb medicines have been used. It is still unclear how the sum of several extracts of anti-thirst drugs represents the anti-lipid peroxidative action. Three anti-thirst herb medicines, Kalopanax pictus (K), Pueraria thunbergiana (P) and Rhus verniciflua (R), were extracted with MeOH and $H_2O$, respectively, and the former one was fractionated into the resultant EtOAc extract. Each extract was reconstituted to give KPR311, KPR131 and KPR113 where, for example, KPR311 represents the complex of K-P-R {3:1:1 (w/w/w)} of the three extracts. The order of the inhibitory effect in bromobenzene-induced lipid peroxidation in rats was as follows: EtOAc extract＞$H_2O$ extract＞MeOH extract. Extract complexes were found to be more potent than the extracts of individual crude drugs. The KPR131 of EtOAc extract was found to be the most potent among the tested samples. These anti-lipid peroxidative effects were also supported by the decrease of aniline hydroxylase activity and aminopyrine N-demethylase activity, on the other hand by the increase of epoxide hydrolase activity. All the tested samples were assayed in vitro antioxidative effects such as DPPH assay, ADP/NADPH/Fe$^{3+}$ assay and ascorbic acid/Fe$^{2+}$ assay. The EtOAc extracts also showed the most significant antioxidative effects. These results suggest that the sum of anti-thirst drugs could reflect the effects of respective crude drugs.s.s.

• Preventive Nutrition and Food Science
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• v.5 no.1
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• pp.48-53
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• 2000

Inhibitory effects of the methanol extract, hexane extract, methanol soluble fraction (MSF) and juice from 3 weeks fermented Kimchi on the tumor formation in sarcoma-180 cell transplanted mice were studied. Effects of the solvent extracts and juice of the Kimchi on the levels of lipid peroxide, glutathione, and the enzyme activities of the liver were also investigated in normal and sarcoma-180 cell transplanted mice. At 32 days following trans-plantation, MSF reduced the tumor formation by 54% compared with the control group, resulting in the smallest tumor weight. Lipid peroxided content in liver increased by the transplantation of sarcoma-180 cells. However, it decreased when MSF of Kimchi was treated to the mice. MSF also suppressed xanthine oxidase activity in cytosol of the liver cells in mice transplanted by sarcoma-180 cells. Kimchi extracts had no inhibitory effect on hepatic aminopyrine-N-demethylase activity in sarcoma-180 cell transplanted or normal mice. Methanol extract and hexane extract of Kimchi slightly increased hepatic glutathione contents in sarcoma-180 treated mice. The injection of MSF from Kimchi markedly increased glutathione levels in the liver of sarcoma-180 treated mice. The injection of MSF from Kimchi markedly increased glutathione levels in the liver of sarcoma-180 treated mice compared to the controls. The MSF recovered the activities of hepatic glutathione reductase and glutathione S-transferase that decreased by the injection of sarcoma-180 cells. These results showed that MSF of Kimchi could suppress the growth of tumors, inhibiting lipid peroxide production and xanthine oxidase activity, in mice. We also suggested that Kimchi extract might play an important role in the prevention of cancer by enhancement of the glutathione level itself as well as via glutathione reductase and glutathione S-transferase.