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Carcinogenicity and mutagenicity of heterocyclic amines in transgenic models

  • Ryu D.Y.
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2000.11a
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    • pp.45-67
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    • 2000
  • 2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) is a mutagenic and carcinogenic heterocyclic amino found in cooked meat. The in vivo mutagenicity and hepatocarcinogenicity of MeIQx were examined in mice harboring the lacZ mutation reporter gene ($Muta^{TM}$ Mice) and bitransgenic mice over-expressing the c-myc oncogene. C57B1/$\lambda$lacZ and bitransgenic c-myc (albumin promoter)/$\lambda$lacZ mice were bred and weaned onto an AIN-76 based diet containing $0.06\%$ (w/w) MeIQx or onto control diet. After 30 weeks on diet, only male bitransgenic mice on MeIQx developed hepatocellular carcinoma ($100\%$ incidence) indicating that there was synergism between c-myc over-expression and MeIQx. By 40 weeks, hepatic tumor incidence was $100\%$ ($17\%$) and $44\%$ ($0\%$) in male c-myc/$\lambda$lacZ and C57B1/$\lambda$lacZ mice given MeIQx (or control) diet, respectively, indicating that either MeIQx or c-myc over-expression alone eventually induced hepatic tumors. At either time point, mutant frequency in the lacZ gene was at least 40-fold higher in MeIQx-treated mice than in control mice of either strain. These findings suggest that MeIQx-induced hepatocarcinogenesis is associated with MeIQx-induced mutations. Elevated mutant frequency in MeIQx-treated mice also occurred concomitant with the formation of MeIQx-guanine adducts as detected by the $^{32}P$-postlabeling assay. Irrespective of strain or diet, sequence analysis of the lacZ mutants from male mouse liver showed that the principal sequence alteration was a single guanine-base substitution. Adenine mutations, however, were detected only in animals on control diet. MeIQx-fed mice harboring the c-myc oncogene showed a l.4-2.6-fold higher mutant frequency in the lacZ gene than mice not carrying the transgene. Although there was a trend toward higher adduct levels in c-myc mice, MeIQx-DNA adduct levels were not significantly different between c-myc/$\lambda$lacZ and C57B1/$\lambda$lacZ mice after 30 weeks on diet. Thus, it appeared that factors in addition to MeIQx-DNA adduct levels, such as the enhance rate of proliferation associated with c-myc over-expression, may have accounted for a higher mutant frequency in c-myc mice. In the control diet groups, the lacZ mutant frequency was significantly higher in c-myc/$\lambda$lacZ mice than in 057B1/$\lambda$1acZ mice. The findings are consistent with the notion that c-myc over-expression is associated with an increase in mutagenesis. The mechanism for the synergistic effects of c-myc over-expression on MeIQx hepatocarcinogenicity appears to involve an enhancement of MeIQx-induced mutations.

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Curcumin and hesperetin attenuate D-galactose-induced brain senescence in vitro and in vivo

  • Lee, Jihye;Kim, Yoo Sun;Kim, Eunju;Kim, Yerin;Kim, Yuri
    • Nutrition Research and Practice
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    • v.14 no.5
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    • pp.438-452
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    • 2020
  • BACKGROUND/OBJECTIVES: Brain senescence causes cognitive impairment and neurodegeneration. It has also been demonstrated that curcumin (Cur) and hesperetin (Hes), both antioxidant polyphenolic compounds, mediate anti-aging and neuroprotective effects. Therefore, the objective of this study was to investigate whether Cur, Hes, and/or their combination exert anti-aging effects in D-galactose (Dg)-induced aged neuronal cells and rats. MATERIALS/METHODS: SH-SY5Y cells differentiated in response to retinoic acid were treated with Cur (1 μM), Hes (1 μM), or a combination of both, followed by 300 mM Dg. Neuronal loss was subsequently evaluated by measuring average neurite length and analyzing expression of β-tubulin III, phosphorylated extracellular signal-regulated kinases, and neurofilament heavy polypeptide. Cellular senescence and related proteins, p16 and p21, were also investigated, including their regulation of antioxidant enzymes. In vivo, brain aging was induced by injecting 250 mg/kg body weight (b.w.) Dg. The effects of supplementing this model with 50 mg/kg b.w. Cur, 50 mg/kg b.w. Hes, or a combination of both for 3 months were subsequently evaluated. Brain aging was examined with a step-through passive avoidance test and apoptosis markers were analyzed in brain cortex tissues. RESULTS: Cur, Hes, and their combination improved neuron length and cellular senescence by decreasing the number of β-gal stained cells, down-regulated expression of p16 and p21, and up-regulated expression of antioxidant enzymes, including superoxide dismutase 1, glutathione peroxidase 1, and catalase. Administration of Cur, Hes, or their combination also tended to ameliorate cognitive impairment and suppress apoptosis in the cerebral cortex by down-regulating Bax and poly (ADP-ribose) polymerase expression and increasing Bcl-2 expression. CONCLUSIONS: Cur and Hes appear to attenuate Dg-induced brain aging via regulation of antioxidant enzymes and apoptosis. These results suggest that Cur and Hes may mediate neuroprotective effects in the aging process, and further study of these antioxidant polyphenolic compounds is warranted.

The Effects of Genistein on the Proliferation and Type I pN Collagen Synthesis in Aged Normal Human Fibroblasts (제니스테인에 의한 노화된 피부세포 활성화와 콜라겐 생성 효과)

  • Yang, Eun-Soon;Hong, Ran-Hi;Kang, Sang-Mo
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.316-324
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    • 2007
  • We studied the effects of genistein obtained from glycolysis of genistin, a kind of phytoestrogen present in soybeans, on cell proliferation and type I pN collagen synthesis in normal human dermal fibroblasts(NHDF). Cell proliferation was increased significantly with genistein treatment at 54-year aged NHDF. Genistein increased cell proliferation more strongly in cells form old doner than young doner. The senescence-associated ${\beta}$-galatosidase activity was decreased in NHDF from 77-year old doner with genistein treatment. Type I pN collagen synthesis was increased with genistein treatement in UVA treated and non-treated NHDF. The increasement of collagen synthesis was more effective in aged cells than young cells. Type I pN collagen synthesis was also increased with genistein treatment in collagen matrix culture with NHDF from sun-exposed and non-exposed skin from 54-year old doner. Genistein treatment inhibited MMP-1 synthesis in old NHDF but not in young NHDF. In conclusion, genistein may be a useful agent for preventing intrinsic aging as well as photoaging.

Tyrosinase Inhibitory Xanthones from Cudrania tricuspidata (꾸지뽕나무에서 분리한 xanthones의 tyrosinase 저해제 연구)

  • Oh, Sang-Seok;Seo, Eun-Jin;Kim, Hoi-Young;Ryu, Young-Bae;Lee, Jin-Hwan;Gal, Sang-Wan;Park, Ki-Hun
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.476-481
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    • 2007
  • The methanolic roots bark extract of Cudrania tricuspidata (Carr.) Bureau was chromatographed, which yielded three xanthones 1-3 by tyrosinase inhibitory activity-guided fractionation. The structures were fully characterized by analysis of physical and spectral data. Among them, furano prenylxanthone 3, never reported as tyrosinase inhibitor, showed potent activity with $IC_{50}$ value of $16.5{\mu}M$, and appeared to inhibit the polyphenol oxidase activity of tyrosinase in an uncompetitive inhibitor($K_i=1.6{\mu}m$) when L-tyrosine was used as a substrate. Moreover, potent inhibitor furano prenylxanthone 3 had an extended lag time of 310 sec at $20{\mu}M$, while lag time of kojic acid as positive control was prolonged with 350 sec at the same concentration.

Ghost Vaccine Prepared from Strong Virulent Salmonella typhimurium Does not Improve Immune Responses of BALB/c Mice (독력이 강한 S. typhimurium으로부터 유도된 고스트 백신으로 면역응답 개선에 관한 연구)

  • Ha, Yeon Jo;Kim, Tae Wan;Kim, Seung Tae;Gal, Sang Wan;Kim, Sam Woong
    • Journal of Life Science
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    • v.24 no.1
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    • pp.39-45
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    • 2014
  • Salmonella typhimurium MMP13 and S. typhimurium ${\chi}8554$ were derived from weak JOL401 and strong ${\chi}3339$ virulent strains. Heat-labile subunit B (LT-B) was used as an adjuvant to increase the effectiveness of the vaccine. Plasmid pMMP184 carrying a ghost cassette was transformed into MMP13 and ${\chi}8554$ to produce the ghost, and the prepared ghost cells were administered into the muscles of BALB/c mice. In the absence of the adjuvant, the total IgG content showed a tendency to increase contrary to the original virulent strength. In contrast, in the presence of the adjuvant, the strain that originated from the strong virulent showed a tendency to promote the immune more than that of weak virulent strain. However, the final concentration of total IgG was similar between the compared groups, indicating that the originated virulent strength does not affect a specific immune. Other elements of the immunoglobulins IgG1, IgG2a, and sIgAs did not show a specific trend. The results of Salmonella challenge showed a similar tendency to regardless of the originated virulence. Taken together, the results suggest that the Salmonella ghost cells promoted the immune system of BALB/c mice, irrespective of the virulence applied to create the ghost cells.

Synthesis and Characterization of Poly(L-lactide)(L-PLA), Poly(D-lactide)(D-PLA) and Stereocomplex-poly(lactide)(PLA) (L-폴리락타이드, D-폴리락타이드의 활성과 입체복합체 폴리락타이드의 제조 및 특성연구)

  • Kim, Ji-Hyun;JeGal, Jong-Geon;Song, Bong-Keun;Shin, Chae-Ho
    • Polymer(Korea)
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    • v.35 no.1
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    • pp.52-59
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    • 2011
  • L-PLA or D-PLA was synthesized in bulk at $140^{\circ}C$ by ring opening polymerization(ROP) of L-lactide or D-lactide as a monomer using tin(II) octoate and lauryl alcohol as a catalyst and an initiator with changing the amounts of catalyst(0.25~1.0 wt%) and initiator(0.l~0.5 wt%). And stereocomplex-PLA was prepared by L-PLA/D-PLA having a wide range of molecular weight(30000~90000 g/mol) and L-PLA/D-PLA blends having different mixing ratio ($X_D$). The melting temperature. thermal degradation temperature and thermal stability of stereocomplex-PLA were higher than those of homopolymers(L-PLA, D-PLA). We supposed that these improvements arose from a strong interaction between L-PLA and D-PLA. The improved mechanical properties and changes in morphology of LPLA/D-PLA blends were compared to those of homopolymers(L-PLA, D-PLA).

Efficient Secretory Expression of Recombinant Endoxylanase from Bacillus sp. HY-20 in Saccharomyces cerevisiae (출아효모(Saccharomyces cerevisiae)에서 Bacillus sp. HY-20균주의 재조합 endoxylanase의 효율적 분비 발현)

  • Kim, Min-Ji;Kim, Bo-Hyun;Nam, Soo-Wan;Choi, Eui-Sung;Shin, Dong-Ha;Cho, Han-Young;Son, Kwang-Hee;Park, Ho-Yong;Kim, Yeon-Hee
    • Journal of Life Science
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    • v.23 no.7
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    • pp.863-868
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    • 2013
  • The XylP gene, which encodes endoxylanase in Bacillus sp. HY-20, was subcloned, and two expression plasmids, pG-xylP and pGMF-xylP were constructed. These plasmids, which contain different signal sequences, XylP s.s and $MF{\alpha}_{opt}$ s.s, respectively, for the secretory expression of endoxylanase, were transformed into Saccharomyces cerevisiae SEY2102 and FY833, respectively. The recombinant endoxylanases were successfully expressed, with a total activity range of 23.7-70.1 unit/ml according to the expression system and host strain. The endoxylanase activity in SEY2102/pGMF-xylP reached a maximum of 88.1 unit/ml in baffled flask culture. Most of the recombinant endoxylanase was efficiently secreted in the extracellular fraction, and the $MF{\alpha}_{opt}$ s.s was more efficient for secreting endoxylanase in yeast than the XylP s.s. Therefore, the expression system developed in this study produces large extracellular amounts of endoxylanase using S. cerevisiae as the host strain, and it could be used in bioethanol production and industrial applications.

Central Neural Pathways Projecting to the Acupoints B62 and K6 Using the Neural Tracer (신경추적자를 이용한 신맥과 조해에서 투사되는 신경원의 표지부위에 대한 연구)

  • Kim, Su-Hyun;Lee, Chang-Hyun;Yuk, Sang-Won;Lee, Kwang-Gyu;Lee, Tae-Young;Lee, Sang-Ryoung
    • Journal of Acupuncture Research
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    • v.18 no.2
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    • pp.51-66
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    • 2001
  • The purpose of this morphological studies was to investigate the central neural pathway projecting to the acupoints $B_{62}$ and $K_6$ using the neuroanatomical method following injection of transsynaptic neurotropic virus, pseudorabies virus(PRV-Ba and PRV-Ga) into the $B_{62}$ and $K_6$. After survival times of 96 hours following injection into the twenty rats with PRV-Ba(Bartha strain) and PRV-Ga(Bartha strain, ${\beta}$-galacidodase insertion). They were perfused, and their spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by X-gal histochemical and PRV immunohistochemical staining method, and observed with light microscope. The results were as follows : 1. In spinal cord, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in thoracic, lumbar and sacral spinal segments. In thoracic spinal segments, Densely labeled areas were founded in lamina IV, V, VII(intermediolateral nucleus) and X areas. In lumbar segemnts, labeled areas were founded in lamina II, IV, V and X areas. In sacral spinal segments, labeled areas were founded in lamina IV, V and VI areas. 2. In brain, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in the $A_1$ noradrenalin cells/$C_1$ adrenalin cells/caudoventrolateral reticular nucleus, rostroventrolateral reticular nuclens, nucleus tractus solitarius, area postrema, raphe obscurus nucleus, raphe paltidus nucleus, raphe magnus nucleus, lateral paragigantoceltular nucleus, lateral rcticular nucleus, gigantocellular nucleus, locus coeruleus, subcoeruleus nucleus, motor trigeminal nucleus, Kolliker-Fuse nucleus, $A_5$ cell group, central gray matter, oculomotor nerve, paraventricular hypothalamic nucleus, median eminence, amygdaloid nucleus, frontal cortex, forelimb area, hindlimb area, 1, 2 areas of parietal cortex and granular and agranular cortex. This results were suggest that overlaped PRV-Ba and PRV-Ga labeled areas projecting to the $B_{62}$ and $K_6$ may be related to the emotional relay pathway in the central autonomic center.

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The Study on the Amount of Trace Elements in Some Fermented Fich Products(jeot-gal) from Some Areas of the West Coast in Korea (서해안 일부지역에서 생산된 젓갈의 무기질 함량조사)

  • 김애정;김순경
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.6
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    • pp.1063-1067
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    • 1997
  • This study was performed to assess the levels of the trace elements(Fe, Cu, Zn, Cr, Co, Mn, Pb, and Cd) in salt-fermented fish products from some areas of the west coast in Korea. Seven samples were Shrimp(Seawoo-jeot), Clam(Jogai-jeot), Oyster(Orikul-jeot), big eyed horring(Bendeng-ie jeot), Mysis(Gonjeng-ie jeot), Hwangandali(Hwangsegi-jeot), and Squid, Han Chi(Han chi-jeot). They were ashed with ternary solution. After ashing the samples, the amount of trace elements in the samples were measured by ICP. The moisture content of the 7 samples before freezing dry were 68.36, 71.52, 81.19, 62.27, 71.30, 64.27, and 66.74%, respectively. Jogai-jeot and Gonjeng-ie jeot contained the most amount of moisture among the samples. Fe contents were 66.46, 309.10, 27.03, 23.01, 132.45, 35.75, and 9.72ppm, respectively. Jogai-jeot contained the most amount of Fe among the samples. Cu contents were 4.60, 4.36, 3.75, 2.21, 10.36, 2.71, and 58.15ppm, respectively. Hanchi-jeot contained the most amount of Cu among the samples. Zn contents were 16.02, 75.06, 37.43, 28.43, 132.45, 35.75, and 9.72ppm, respectively. Gonjeng-ie jeot contained the most amount of Zn among the samples. Cr contents were 0.80, 1.61, 0.84, 0.96, 1.12, 0.96, and 0.59ppm, respectively. Jogai-jeot contained the most amount of Cr among the samples. Co contents were 0.13, 0.54, 0.31, 0.46, 0.50, 0.63, and 0.35ppm, respectively. Hwangsegi-jeot contained the most amount of Co among the samples. Mn contents were 7.30, 10.69, 14.87, 4.12, 8.03, 2.94 and 1.54ppm, respectively. Origkul-jeot contained the most amount of Mn among the samples. Pb contents were 1.80, 4.30, 2.53, 4.61, 3.08, 5.04, and 2.74ppm, respectively. Hwangsegi-jeot contained the most amount of Pb among the samples. Cd contents were 0.005, 0.03, 0.06, 0.005, 0.01, 0.00, and 0.10ppm, respectively. Hanchi-jeot contained the most amount of Cd among the samples. This study is limited within 7 samples caught and producted from the some areas of the west coast in Korea. Therefore, I hope there will be broader experiments concerned with this study to make clear not only nutritional aspect(the contents of Fe, Cu, Zn, Cr, Co, and Mn) but also toxicological aspect(the contents of Pb and Cd).

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Antioxidant Properties of Peptides Extracted from Tenebrio molitor Larvae (갈색거저리 유충에서 추출한 펩타이드의 항산화 특성)

  • Sam Woong Kim;Sang Wan Gal;Won-Jae Chi;Woo Young Bang;So Jeong Park;Tae Wan Kim;Kyu Ho Bang
    • Journal of Life Science
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    • v.33 no.5
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    • pp.383-390
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    • 2023
  • The goal of this study was to identify new bioactive peptides in extracts derived from Tenebrio molitor (T. molitor) larvae for the development of functional foods. After extraction from freeze-dried T. molitor larvae with various solvents on time course, the extracts showed the highest 2,2,1-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity at 5 and 10 hr per total protein and solid contents, respectively. When the water extract was fractionated, a high methanol concentration led to a reduced level of high-molecular-weight proteins in the centrifugal supernatant, whereas increased DPPH activity in the supernatants suggests low-molecular-weight peptides may mediate antioxidant activity in the supernatant. Most of the organic solvent partitions, excluding butanol, showed similar activities in the water phases, and the organic solvent partition fraction exhibited a 28~44% decrease in activity following heat treatment, implying that some components in the fraction become unstable in the presence of heat. The addition of proteinase K to the water extract increased DPPH activity by 10~20%, suggesting that peptides, when released from total proteins, partially increase antioxidant activity. Therefore, we suggest that the antioxidants in T. molitor larval extracts make them a potential source of functional animal food.