• Korean Journal of Weed Science
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• v.3 no.1
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• pp.75-99
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• 1983

The herbicidal properties of perfluidone [1,1,1-trifluoro-N-2-methyl-4-(phenylsulponyl) phenyl methanesulfonamide] were investigated in pots and paddy fields. At the rate of 2.0kg prod./10a, perfluidone did not cause any injury to the 4 leaf stage (LS) rice seedlings. Although the crop injury increased with increasing the application rate, the injury caused by 16kg prod. perfluidone/10a gave rise to only 30% yield reduction. The crop injury was greatest when perfluidone was applied 2 days before transplanting and decreased as the application time delayed. Perfluidone showed greater crop injury to the 3 LS seedlings, at more than 7cm water depth, and at high temperature than to the 4 LS seedlings, at 3-5cm water depth, and at low temperature. Indica and indica ${\times}$ japonica rice varieties were generally more sensitive to perfluidone than japonica rice variety. Perfluidone effectively controlled most of annual weeds and such perennial weeds as Sagittaria pygmaea MIQ., Potamogeton distinctus A. BENN, Cyperus serotinus ROTTB, Scirpus maritimus L., Eleocharis kuroguwai OHWL, and Scirpus hotarui OHWL, whereas Sagittaria trifolia L. and Polygonum hydropiper SPACH. were tolerent to perfluidone. The weeding effect decreased with increasing the leaching amount of water and the overflowing of irrigated water within 24 hours after the herbicide application. When the application time was done later than 8 days after transplanting, the perennial weeds were shown at deeper soil layers, and the standing water was deeper than 7cm, the effect tended to decrease. However, there was no difference in the weeding effect between soil types. Downward movement of perfluidone in flooded soil ranged from 2 to 8cm deep. The movement increased with increasing the leaching amount of water and the application rate and at a sandy loam soil which possessed less adsorptive capacity. Residual effect of perfluidone was found at 35 to 80 days after application, which varied such factors as Soil types. Increase in the leaching amount of water resulted in decrease in the period of the residual effect. The period was shorter at non-sterilized soil than at sterilized soil. The 0.75kg ai perfluidone + 1.5kg ai SL-49 (1,3-dimethyl-6-(2,4-dichlor-benzoyl)-5-phenacyloxy-pyrazole)/ha and 1.5kg ai perfluidone + 1.05kg ai bifenox (2,4-dichlorophenyl-3-methoxy carbonyl-4-nitro phenyl ether)/ha showed less crop injury than 1.5kg ai/ha perfluidone alone. However, the weeding effect of the former was similar to that of the later.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.63 no.1
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• pp.25-34
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• 2018

This study was conducted to investigate the germination and proteome profile characteristics of wheat seeds treated under various concentrations of abscisic acid (ABA). After-ripening, the seeds of three wheat cultivars (Baegjoong, Keumkang, and Uri) showing different levels of dormancy were used. Germination index and germination rate of the cultivars was higher than 0.95% and 98%, respectively, and these were not significantly different under 0, 10, 30, and $50{\mu}M$ ABA at 7 d after germination. However, the growth of the shoot and radicle was significantly inhibited at 10, 30, and $50{\mu}M$ ABA compared to that at $0{\mu}M$ ABA. Mean ABA content of the embryos of seeds germinated at 0 and $50{\mu}M$ ABA for 7 d was 0.8 and $269.0ngmg^{-1}DW$, respectively. Proteins extracted from embryos germinated for 4 d were analyzed by two-dimensional gel electrophoresis, and proteins showing a difference of 1.5-fold or greater in their spot volume relative to that of $0{\mu}M$ ABA were identified. The expression of four protein spots increased at $50{\mu}M$ ABA and two protein spots were detected only at $50{\mu}M$ ABA; these six proteins were all identified as globulin types. Conversely, the expression of three protein spots decreased at $50{\mu}M$ ABA and were identified as cytosolic glutamine sysnthetase, isocitrate dehydrogenase, and S-adenosylmethionine synthetase 2. In conclusion, ABA did not inhibit the germination rate regardless of pre-harvest sprouting characteristics of the cultivars. However, the growth of the shoot and radicle was significantly inhibited by ABA, most likely through the down regulation of glutamine, methyl group donor, and polyamines biosynthesis, among others, while accompanied by globulin accumulation in the embryos.

• The Korean Journal of Pharmacology
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• v.32 no.3
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• pp.407-416
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• 1996

The reactive intermediates formed during the metabolism of therapeutic agents, toxicants and carcinogens by cytochromes P450 are frequently capable of covalently binding to tissue macromolecules and causing tissue damage. It has been shown that YH439, a congener of malotilate, is effective in suppressing hepatic P450 2E1 expression. The present study was designed to further establish the mechanistic basis of YH439 protection against toxicant by assessing its effects against chemical-mediated potentiated hepatotoxicity. Retinoyl palmitate (Vit-A) pretreatment of rats for 7 days substantially enhanced carbon tetrachloride hepatotoxicity, as supported by an ${\sim}5-fold$ increase in serum alanine aminotransferase (ALT) activity, as compared to $CCl_4$ treatment alone. The elevation of ALT activity due to Vit-A was completely blocked by the treatment of $GdCl_3$ a selective inhibitor of Kupffer cell activity. Concomitant pretreatment of rats with both YH439 and Vit-A resulted in a 94% decrease in Vit-A-potentiated $CCl_4$ hepatotoxicity. YH439 was also effective against propyl sulfide-potentiated $CCl_4-induced$ hepatotoxicity. Whereas propyl sulfide (50 mg/kg, 7d) enhanced $CCl_4-induced$ hepatotoxicity by >5-fold, relative to $CCl_4$ treatment alone, concomitant treatment of animals with both propyl sulfide and YH439 at the doses of 100 and 200 mg/kg prevented propyl sulfide-potentiated $CCl_4$ hepatotoxicity by 35% and 90%, respectively. Allyl sulfide, a suppressant of hepatic P450 2E1 expression, completely blocked the propyl sulfide-enhanced hepatotoxicity, indicating that propyl sulfide potentiation of $CCl_4$ hepatotoxicity was highly associated with the expression of P450 2E1 and that YH439 blocked the propyl sulfide-enhanced hepatotoxicity through modulation of P450 2E1 levels. Propyl sulfide- and $CCl_4-induced$ stimulation of lipid peroxidation was also suppressed by YH439 in a dose-related manner, as supported by decreases in malonedialdehyde production. The role of P450 2E1 induction in the potentiation of $CCl_4$ toxicity and the effects of YH439 were further evaluated using pyridine as a P450 2E1 inducer. Pyridine pretreatment substantially enhanced the $CCl_4$ hepatotoicity by 23-fold, relative to $CCl_4$ alone. YH439, however, failed to reduce the pyridine-potentiated toxicity, suggesting that the other form(s) of cytochroms P450 inducible by pyridine, but not suppressible by YH439 treatment, may play a role in potentiating $CCl_4-induced$ hepatotoxicity. YH439 was capable of blocking cadmium chloride-induced liver toxicity in mice. These results demonstrated that YH439 efficiently blocks Vit-A-enhanced hepatotoxiciy through Kupffer cell inactivation and that the suppression of P450 2E1 expression by YH439 is highly associated with blocking of propyl sulfide-mediated hepatotoxicity.

• Journal of Nutrition and Health
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• v.43 no.6
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• pp.588-596
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• 2010

Most nutrients taken by pregnant women are secreted into their breast milk. Food contains choline together with betaine, and in human body choline is oxidized to betaine which transfer methyl group. The aim of the study was to estimate the concentrations of choline and betaine in breast milk of Korean lactating women and the choline and betaine intakes of their infants. Total choline, free choline and betaine concentrations in breast milk of some lactating women living in Daejon Metropolitan city were analyzed every month by using HPLC-MS and enzymatic method during the first five months. Total choline concentrations of breast milks were 157.64 mg/L (1.52 mmol/L), 157.83 mg/L (1.52 mmol/L), 165.99 mg/L (1.60 mmol/L), 153.67 mg/L (1.48 mmol/L), 145.05 mg/L (1.39 mmol/L) by month after delivery for five months. The concentrations of total choline and free choline in breast milks were not significantly changed for the five months while the betaine concentrations gradually decreased. Daily intake of total choline of the infants appears to be adequate for the infant's requirement according to the US DRI; 124.6 mg/d, 120.9 mg/d, 126.5 mg/d 104.1 mg/d from 2nd to 5th month after birth. Free choline and betaine intakes of the infants were not significantly changed during the four months except showing decrease in betaine intake per kg body weight. Choline intakes of the infants more correlated with choline concentrations of the breast milks (r = 0.982, p = 0.000) than intake amount of the breast milk (r = 0.414, p = 0.028). These results suggest that the choline intake of Korean breast-fed infants appears to be adequate and the intake could be affected by the choline concentration of the breast milk.

• Applied Microscopy
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• v.40 no.2
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• pp.53-64
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• 2010

Glutamate receptors may play a critical role in the refinement of developing synapses. The lateral superior olivary nucleus (LSO)-medial nucleus of trapezoid body (MNTB) synaptic transmission in the mammalian auditory brain stem mediate many excitatory transmitters such as glutamate, which is a useful model to study excitatory synaptic development. Hearing deficits are often accompanied by changes in the synaptic organization such as excitatory or inhibitory circuits as well as anatomical changes. Owing to this, circling mouse whose cochlea degenerates spontaneously after birth, is an excellent animal model to study deafness pathophysiology. However, little is known about the development regulation of the subunits composing these receptors in circling mouse. Thus, we used immunohistochemical method to compare the N-Methyl-D-aspartate receptor (NMDA receptor) NR1, NR2A, NR2B distribution in the LSO which project glutamergic excitatory input into the auditory brainstem, in circling mouse of postnatal (p) 7 and 16, which have spontaneous mutation in the inner ear, with wild-type mouse. The relative NMDAR1 immunoreactive density of the LSO in circling mouse p7 was $128.67\pm8.87$ in wild-type, $111.06\pm8.04$ in heterozygote, and $108.09\pm5.94$ in homozygote. The density of p16 circling mouse was $43.83\pm10.49$ in wild-type, $40\pm13.88$ in heterozygote, and $55.96\pm17.35$ in homozygote. The relative NMDAR2A immunoreactive density of LSO in circling mouse p7 was $97.97\pm9.71$ in wild-type, $102.87\pm9.30$ in heterozygote, and $106.85\pm5.79$ in homozygote. The density of LSO in p16 circling was $47.4\pm20.6$ in wild-type, $43.9\pm17.5$ in heterozygote, and $49.2\pm20.1$ in homozygote. The relative NMDAR2B immunoreactive density of LSO in circling mouse p7 was $109.04\pm6.77$ in wild-type, $106.43\pm10.24$ in heterozygote, and $105.98\pm4.10$ in homozygote. the density of LSO in p16 circling mouse was $101.47\pm11.5$ in wild-type, $91.47\pm14.81$ in heterozygote, and $93.93\pm15.71$ in homozygote. These results reveal alteration of NMDAR immunoreactivity in LSO of p7 and p16 circling mouse. The results of the present study are likely to be relevant to understand the central change underlying human hereditary deafness.

• Journal of the Korean Applied Science and Technology
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• v.18 no.3
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• pp.197-213
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• 2001

Conjugate trienoic acids (CTA) occurred in triacylglycerols (TGs) of the seed oils of Trichosanthes kirilowii, Momordica charantia and Aleurites fordii, and they were easily converted to their methyl esters in a mixture of sodium methoxide-methanol without any structural destruction. The main fatty acids in triacylglycerol (TG) fraction of the seed oils of Trichosanthes kirilowii are $C_{18:2{\omega}6}$ (32.2 mol %), $C_{18:3{\;}9c.11t,13c}$ (38.0 mol %) and $C_{18:1{\omega}9}$ (11.8 mol %), followed with $C_{16:0}$ (4.8 mol %) and $C_{18:0}$ (3.1 mol %). The TG fraction was resolved into 20 TG molecular species according to the partition number (PN) by reversed-phase (RP)-HPLC. The main TG species were $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$, of which amounts reached 63 mol % of total TG molecular species. The TG sample was fractionated into 11 fractions according to the number of double bond in the molecule by $Ag^{+}-HPLC$ and the species of $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$ were also eluted as main components. The TG species containing CTA showed unusual behaviours in the order of elution by HPLC ; first, TG moleular species of $DT_{c2}$ (D; dienoic acid, $T_{c}$; punicic acid, $T_{ci}$; ${\alpha}-eleostearic$ acid, M ; monoenoic acid, $S_{t}$; stearic acid) was eluted earlier than $Mt_{c2}$, although they have the same PN number of 40, and, secondly, the species of $DT_{ci2}$ with eight double bonds was eluted earlier than that of $D_2T_{ci}$ with seven double bonds. Intact TG of the seed oils of Momordica charantia contained mainly fatty acids such as $C_{18:3{\omega}9c,11t,13t}$ (57.7 mol %), $C_{18:1{\omega}9}$ (17.4 mol %), $C_{18:0}$ (12.3 mol %) and $C_{18:2{\omega}6}$ (10.6 mol %), and was classified into 13 fractions by RP-HPLC. The main TG species were as follows ; $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.1 mol %] and $S_{t}T_{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_2$, 33.9 mol %] comprising about 73 mol % of total TG species, accompanied by $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 7.3 mol %], $D_{2}T_{ci}$ [$ (C_{18:2{\omega}6})_{2}(C_{18:3\;9c,11t,13t})$, 3.6 mol %] and $MDT_{ci}$ [$(C_{18:1{\omega}9})(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})$, 3.5 mol %]. Simple TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_3]$ was present in a small amount of 1.4 mol %, but other simple TG species were not detected. The TG was also resolved into 11 fractions according to the number of double bond by $Ag^{+}-HPLC$, and the species were mainly occupied by $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.4 mol %] and $S_tT-{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_{2}$, 35.4 mol %] $DT_{ci2}$ species with eight double bonds was also developed faster than $D_2T_{ci}$ one with seven double bonds as indicated in the analysis of TG of the seed oils of T. kirilowii, and $MT_{ci2}$ species with cis, trans, trans-configurated double bond was eluted earlier than $MT_{c2}$ species with cis, trans, cis-configurated double bond. The main components of fatty acid in total TG fraction isolated from the seed oils of of Aleurites fordii were in the following order ; $C_{18:3\;9c,11t,13t}$ (81.2 mol %)> $C_{18:2{\omega}6}$ (8.5 mol %)> $C_{18:1{\omega}9}$ (5.4 mol %)$. With resolution of the TG by RP-HPLC, eight fractions such as $T_{ci3}$, $Dt_{ci2}$, $D_{2}T_{ci}$, $MT_{ci2}$, $PT_{ci2}$ (P; palmitic acid), $PMT_{ci}$, $PDT_{ci}$ and $S_{t}T_{ci2}$ ($S_{t}$; stearic acid) were isolated, respectively. TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_{3}$, 54.2 mol %], $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 15.0 mol %] and $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3 9c,11t,13t})_{2}$, 14.8 mol %] were present as main species.

• Proceedings of the Ginseng society Conference
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• 1980.09a
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• pp.87-113
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• 1980

This experiment was carried out to evaluate the effects of Korean ginseng extract on carcinogenesis induced by various chemical carcinogens. Red ginseng extract was used for this study and was administered orally to the experimental animals. Carcinogens that were injected in subscapsular region of ICR newborn mice within 24 hours after birth were 9,10-dimethyl-1,2-benzan-thracene (DMBA), urethane, N-2-fluorenylacetamide(AAF), aflatoxin $B_1$ and tobacco smoke condensate. N -methyl-N -nitroso-N'-nitroguani-dine(MNNG) was injected subcutaneously at the back of wistar rats. Experimental animals were autopsied in immediately after being sacrificed. All major organs were examined grossly and weighted. After fixation histopathological preparations were made for microscopical study. Following results were obtained. In DMBA group sacrificed at the 26th week after the treatment with DMBA, the incidence of lung adenoma was $77\%$ and the average number of the tumor was 17. However, in DMBA combined with red ginseng group, the incidence was $78\%$ and the average number of lung adenoma was 14.1. This indicates that ginseng extract had no effect on the incidence of lung adenoma but decreased the average number of lung adenoma by $17\%.$ In DMBA group sacrificed at the 48th week after the injection of DMBA, the lung adenoma incidence was $88\%.$ The average diameter of the largest lung adenoma was 3.5 cm, the incidence of diffuse pulmonary infiltration was $18\%$ and the average lung weight of male experimental mice was $528.2{\pm}469.1\;gm.$ On the other hand, in DMBA combined with red ginseng group sacrificed at the 48th week, the incidence of lung adenoma was $96\%.$ The average diameter of the largest adenoma was 2.7 cm, the incidence of diffuse pulmonary infiltration was $7\%$ and the average lung weight of male mice was $418.0{\pm}520\;gm.$ These observations show that ginseng extract did not have any inhibitory effect on the incidence of lung adenoma but decreased the average diameter of the largest lung adenoma by $23\%,$ the incidence of duffuse pulmonary infiltration by $63\%$ and the average lung weight of male experimental mice by $21\%.$ From these results we have found that the prolonged administration with ginseng extract showed no inhibitory effect on the incidence of adenoma but it had the inhibitory effect on the proliferation of lung adenomas induced by DMBA. In urethane group sacrificed at the 28th week after the injection of urethane, the incidence of lung adenoma was $94\%$ and the average number of lung adenoma was 8.6. In urethane combined with red ginseng group, the. incidence of lung adenoma was $73\%$ and the average number of adenoma was 6.0. These results indicate that there were $22\%$ decrease of the lung adenoma incidence and $31\%$ decrease of the average number of adenoma in urethane combined with red ginseng group. And in urethane group sacrificed at the 50th week, the incidence of lung adenoma was $98\%$ and the incidence of diffuse pulmonary infiltration was $14\%$. In urethane combined with ginseng group the incidence of lung adenoma was $85\%$ and the incidence of diffuse pulmonary infiltration was $12\%$. Therefore the ginseng administration resulted in $15\%$ decrease of the lung adenoma incidence and $14\%$ decrease of the diffuse pulmonary infiltration incidence. From these results we knew that the prolonged administration with ginseng extract inhibited the incidence and also the proliferation of the lung adenoma induced by urethane. Lung adenoma and hepatoma were induced in the experimental mice sacrificed at the 68th week but not in the experimental mice sacrificed at the 28th week after the injection of AAF. In AAF group sacrificed at the 68th week after the injection of AAF the incidence of lung adenoma was $18\%$ and the incidence of hepatoma was $27\%$. And in AAF combined with ginseng group the lung adenoma incidence was $12\%$ and the hepatoma incidence was $37\%$. So the ginseng seemed to decrease the lung adenoma incidence by AAF, but we were unable to conclude the significant inhibitory effect of the ginseng extract on the incidence of lung adenoma by AAF because the above incidence of lung adenoma were similar to that of control group which was $11\%$. And these experimental data revealed that ginseng extract didn't have any inhibitory effect on the incidence of hepatoma induced by AAF. In aflatoxin $B_1$ group sacrificed at the 56th week, the incidence of lung adenoma was $24\%$ and hepatoma was $11\%$. However in aflatoxin $B_1$ combined with ginseng group the incidence of lung adenoma was $17\%$ and hepatoma was $3\%$ These results indicate that there were $29\%$ decrease of the lung adenoma incidence and $75\%$ decrease of the hepatoma incidence in aflatoxin $B_1$ combined with ginseng group. In tobacco smoke condensate experimental group sacrificed at 67th week, no tumors were induced except just a few lung adenoma. The lung adenoma incidence both in tobacco smoke condensate group and in tobacco smoke condensate combined with ginseng group was $8\%$. And this incidence rate was similar to that of control group. These results indicate that the injection of 320 ug tobacco smoke condensate per ICR newborn mouse was unable to induce lung adenoma in our experiments. In MNNG group sacrificed at the 27th week the tumor incidence was $38.5\%$ and in MNNG combined with ginseng extract group was $37\%$. In MNNG group for investigation of the life span of tumor bearing rats the tumor incidence was $93\%$ and the average life span of tumor bearing rats was 318 days. And in MNNG combined with ginseng extract group the tumor incidence was $96\%$ and the average life span was 337 days. Tumor induced by MNNG was almost sarcoma. This indicates that there was no inhibitory effect of ginseng extract on the tumor incidence, but the extract prolonged the average life span of tumor bearing rats by approximately 19 days.

• Journal of Life Science
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• v.17 no.6 s.86
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• pp.791-797
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• 2007

We studied the inhibitory effect of solvent fractions from doenjang on mutagenicity using Salmonella typhimurium TA100 in Ames test. We also investigated the effect of solvent fractions from doenjang on the growth of tumor cells and the production of interleukin-2 (IL-2). The treatment of dichlorormethane and ethylacetate fractions (2.5 mg/assay) from doenjang to Ames test system inhibited aflatoxin B$_1$ (AFB$_1$) induced mutagenicity by 96% and 97%, respectively, and showed a higher antimutagenic effect than other solvent fractions. In case of N-methyl-N'-nitro-N-nitrosoguamidine (MNNG) induced mutagenicity, the ethylacetate fraction showed the highest inhibitory effect (by 75%) among the other sol-vent fractions, although the inhibitory effect was not stronger compared to AFB$_1$ induced mutagenicity. The treatment of dichloromethane and ethylacetate fractions markedly inhibited the growth of Yac-1 (by 80% and 94%, respectively) and sacroma-180 cancer cells (by 60% and 96%, respectively) after 4 days of incubation at 37${\circ}$C. To elucidate the immunological mechanism of antitumor activity of doenjang, spleen cells of Balb/c mouse were exposed to the dichloromethane and ethyl-acetate fractions for 24 hours at 37${\circ}$C . The culture supernatants following the treatment of djchloromethane and ethylacetate factions to spleen cells increased the production of IL-2. These results indicated that the anticarcinogenic effect of doenjang was mediated by the production of IL-2.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.783-794
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• 1996

It has been suggested that ion transport systems are intimately involved in mediating the effects of growth regulatory factors on the growth of a number of different types of animal cells in vivo. The functional importance of the apical membrane $Na^+/H^+$ antiporter in the renal proximal tubule is evidenced by estimates that this transporter mediates the reabsorption of approximately one third of the filtered load of sodium and the bulk of the secretion of hydrogen ions. This study was designed to investigate the pathway utilized by IGF-I in regulating sodium transport in primary cultured renal proximal tubule cells. Results were as follows : 1. $Na^+$ was observed to accumulate in the primary cells as a function of time. Raising the concentration of extracellular NaCl induced an decrease in $Na^+$ uptake compared with control cells in a dose dependent manner. The rate of $Na^+$ uptake into the primary cells was about two times higher in the absence of NaCl($40.11{\pm}1.76pmole\;Na^+/mg\;protein/min$) than in the presence of 140mM NaCl($17.82{\pm}0.94pmole\;Na^+/mg\;protein/min$) at the 30 minute uptake. 2. $Na^+$ uptake was inhibited by IAA($1{\times}10^{-4}M$) or valinomycin($5{\times}10^{-6}M$) treatment($50.51{\pm}4.04$ and $57.65{\pm}2.27$ of that of control, respectively). $Na^+$ uptake by the primary proximal tubule cells was significantly increased by ouabain($5{\times}10^{-5}M$) treatment($140.23{\pm}3.37%$ of that of control). When actinomycin D($1{\times}10^{-7}M$) or cycloheximide($4{\times}10^{-5}M$) was applied, $Na^+$ uptake was decreased to $90.21{\pm}2.39%$ or $89.64{\pm}3.69%$ of control in IGF-I($1{\times}10^{-5}M$) treated cells, respectively. 3. Extracellular cAMP decreased $Na^+$ uptake in a dose-dependent manner($10^{-8}-10^{-4}M$). IBMX($5{\times}10^{-5}M$) also inhibited $Na^+$ uptake. Treatment of cells with pertussis toxin(50pg/ml) or cholera toxin($1{\mu}g/ml$) inhibited $Na^+$ uptake. Extracellular PMA decreased $Na^+$ uptake in a dose-dependent manner(1-100ng/ml). 100 ng/ml PMA concentration significantly inhibited $Na^+$ uptake in IGF-I treated cells. However, staurosporine($1{\times}10^{-7}M$) had no effect on $Na^+$ uptake. When PMA and staurosporine were added together, the inhibition of $Na^+$ uptake was not observed. In conclusion, sodium uptake in primary cultured rabbit renal proximal tubule cells was dependent on membrane potentials and intracellular energy levels. IGF-I stimulates sodium uptake through mechanisms that involve some degree of de novo protein and/or RNA synthesis, and cAMP and/or PKC pathway mediating the action mechanisms of IGF-I.

• Korean journal of applied entomology
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• v.13 no.3 s.20
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• pp.105-133
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• 1974

The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.