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Effect of Extraction Conditions on in vitro Antioxidant Activities of Root Bark Extract from Ulmus pumila L. (추출조건에 따른 유근피 추출물의 항산화 활성)

  • Kim, Jae-Min;Cho, Myoung-Lae;Seo, Kyu-Eun;Kim, Ye-Seul;Jung, Tae-Dong;Kim, Young-Hyun;Kim, Dan-Bi;Shin, Gi-Hae;Oh, Ji-Won;Lee, Jong Seok;Lee, Jin-Ha;Kim, Jong-Yae;Lee, Dae-Won;Lee, Ok-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.8
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    • pp.1172-1179
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    • 2015
  • This study investigated optimal extraction conditions for application of Ulmus pumila L. as a natural antioxidant. U. pumila L. was extracted using ethanol (EtOH) at various concentrations (0, 40, and 80%) and extraction times (1, 2, and 3 h) at $70^{\circ}C$ and then evaluated for extraction yield, total phenolic contents, total flavonoid contents, as well as antioxidant activities [2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, reducing power, and oxygen radical absorbing capacity (ORAC)]. Antioxidant activities were correlated with total phenolic and flavonoid contents. Of the solvent conditions, 80% EtOH extracts for 3 h at $70^{\circ}C$ showed the highest total phenolic and flavonoid contents with strong antioxidant activities, although there were no significant time effects on DPPH and ABTS radical scavenging activities and reducing power. However, ORAC values of all EtOH extracts remarkably increased in a time-dependent manner. In addition, 80% EtOH extract for 3 h exhibited strong antioxidant effects on HDF and 3T3-L1 cells. Therefore, the antioxidant capacity of U. pumila L., may due to phenolic and flavonoid contents, and extraction conditions were 80% EtOH for 3 h at $70^{\circ}C$. This extract could be a good source for natural antioxidants.

Synergistic Inhibition of Aronia melanocarpa and Moringa oleifera Seed Extract on Experimental Atopic Dermatitis (아로니아 및 모링가 종자 복합물의 항아토피 상승효과)

  • Ki, Hyeon-Hui;Lee, Ji-Hyun;Moon, Kwang-Hyun;Lee, Jeong-Ho;Kim, Dae-Geun;Jeong, Kyung-Ok;Im, So-Yeon;Lee, Young-Mi;Kim, Dae-Ki
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.3
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    • pp.298-305
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    • 2017
  • Atopic dermatitis is a chronic, relapsing inflammatory skin disease. This study aimed to investigate the therapeutic benefits of Aronia melanocarpa (AM) and Moringa oleifera seed extract (MO) on experimental atopic dermatitis. We examined the effects of AM or MO and their combination on 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis in BALB/c mice as well as tumor necrosis factor $(TNF)-{\alpha}$ and interferon $(IFN)-{\gamma}-stimulated$ HaCaT keratinocytes. Mice were orally treated with extract during repeated application of DNCB to shaved dorsal skin. Our results show that treatment with AM and MO in combination reduced histological manifestations such as epidermal hyperplasia and inflammatory cell infiltration. Furthermore, it significantly decreased skin thickness and serum immunoglobulin E (IgE) level compared to the AM or MO alone treated group. Combined extract of AM and MO suppressed expression of $TNF-{\alpha}/IFN-{\gamma}-induced$ T helper 2 (Th2) chemokines such as thymus and activation-regulated chemokine and macrophage-derived chemokine. To sum up, combination of AM and MO suppressed the inflammatory response and serum IgE as an indicator of several allergic diseases in DNCB-induced experimental atopic dermatitis and Th2 chemokine expression in HaCaT cells. This result suggests that combination of AM and MO could be a valuable strategy to improve atopic dermatitis.

β-Galactosidase-catalyzed Synthesis of 1, 2-Hexanediol Galactoside and its Purification using Ethyl Acetate Extraction followed by Silica Gel Chromatography (대장균 β-Galactosidse를 이용한 1, 2-Hexanediol galactoside의 합성과 Ethyl Acetate 추출 및 Silica Gel Chromatography를이용한 정제)

  • Kim, Yi-Ok;Jung, Kyung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.3
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    • pp.498-506
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    • 2016
  • 1, 2-Hexanediol galactoside (HD-gal) has been previously synthesized from 1, 2-hexanediol (HD), in which recombinant ${\beta}$-galactosidase (${\beta}$-gal) of Escherichia coli (E. coli) was used for transgalactosylation reaction. In this study, a method for HD-gal purification from the reaction mixture was particularly investigated. Using ${\beta}$-gal-containing E. coli, HD-gal was synthesized from 75 mM HD for 48 hr under 300 g/l lactose concentration. Then, HD-gal synthesis from HD was confirmed by TLC analysis, and the existence of E. coli ${\beta}$-gal during 48 hr-reaction was also confirmed by Western blotting, in which the conversion yield of HD to HD-gal reached about 94% during 48 hr. To establish an efficient method for HD-gal purification, we carried out the solvent extraction of the reaction mixture, followed by silica gel chromatography, particularly in order to remove the residual HD. Two water-immiscible solvents, such as methylene chloride and ethyl acetate, were investigated comparatively to find out appropriate solvent. Then, it was found that residual HD was almost removed when ethyl acetate extraction of water phase of reaction mixture was carried out four times. Subsequently, silica gel chromatography was carried out, and purified HD-gal could be finally obtained. The production yield for HD-gal from 75 mM HD was $8.9{\pm}0.6%$ (n=3) (mole basis) or $21.1{\pm}1.4%$ (n=3) (weight basis). For further study, using purified HD-gal, we will investigate the minimum inhibitory concentrations (MICs) of HD-gal against bacteria. In addition, cytotoxicity to human skin cells of HD-gal will be examined.

NMR Spectroscopy and Mass Spectrometry of 1, 2-Hexanediol Galactoside synthesized using Escherichia coli β-Galactosidase (대장균 베타-갈락토시데이즈를 이용하여 합성된 1, 2-Hexanediol Galactoside의 NMR Spectroscopy 및 Mass spectrometry)

  • Kim, Yi-Ok;Lee, Hyang-Yeol;Jung, Kyung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.2
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    • pp.286-292
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    • 2016
  • 1, 2-Hexanediol galactoside (HD-gal) has been synthesized from 1, 2-hexanediol (HD), a cosmetic preservative, using recombinant Escherichia coli ${\beta}$-galactosidase (${\beta}$-gal) at the high lactose concentration (300 g/l). To confirm the molecular structure of synthesized HD-gal, NMR ($^1H$- and $^{13}C$-) spectroscopy and mass spectrometry of HD-gal were conducted. $^1H$ NMR spectrum of HD-gal showed multiple peaks corresponding to the galactocyl group, which is an evidence of galactocylation on HD. Downfield proton peaks at ${\delta}_H$ 4.44 ppm and multiple peaks from ${\delta}_H$3.96~3.58 ppm were indicative of galactocylation on HD. Up field proton peaks at ${\delta}_H$ 1.60~1.35 ppm and 0.92 ppm showed the presence of $CH_2$ and $CH_3$ protons of HD. $^{13}C$ NMR spectrum revealed the presence of 21 carbons suggestive of ${\alpha}$- and ${\beta}$-anomers of HD-gal. Among 12 carbon peaks from each anomers, the 3 peaks at dC 68.6, 60.9 and 13.2 ppm were assigned to be overlapped showing only 21 peaks out of total 24 peaks. The mass value (protonated HD-gal, m/z = 281.1601) from mass spectrometry analysis of HD-gal, and $^1H$ and $^{13}C$ NMR spectral data were in well agreement with the expecting structure of HD-gal. For further study, the minimum inhibitory concentrations (MICs) of HD-gal against bacteria will be investigated, and, in addition, cytotoxicity to human skin cells of HD-gal will be examined. It is expected that it will eventually be able to develop a new cosmetic preservative, which have low cytotoxicity against human skin cell and maintains antimicrobial effect.

Effect of Resistant Starch on the Large Bowel Environment and Plasma Lipid in Rats with Loperamide-Induced Constipation (저항전분 투여가 Loperamide 유도 변비 쥐의 대장환경과 혈청지질에 미치는 효과)

  • Sin, Hyun-Ju;Kim, Kwang-Ok;Kim, Sung-Hong;Kim, Young-Ah;Lee, Hye-Sung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.5
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    • pp.684-691
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    • 2010
  • The present study was conducted to evaluate the effect of resistant starch (RS) on the large bowel function and plasma lipids in rats with constipation induced by Loperamide. Animals were divided into six groups: normal control-5% cellulose, constipation-5% cellulose, constipation-5% pectin, constipation-5% RS-type 2 (RS2), constipation-8% RS2 and constipation-5% RS type 3 (RS3) groups, and fed experimental diets for five weeks. The results from RS groups were compared with those from other dietary fiber groups. The groups supplemented with RS3 or high level of RS2 showed significantly increased counts of bifidobacteria in the cecum than the other groups. The production of total short chain fatty acids in the cecal contents was significantly high in pectin, RS3 and high RS2 groups. The pH in the cecal contents of the RS supplemented groups was significantly decreased compared with the cellulose supplemented groups. The production of prostaglandin E2 in the colon mucus of the RS groups was higher than the normal group; however, it was significantly decreased compared to the cellulose or pectin supplemented constipated groups. The thickness of the mucus layer and the production of mucus from epithelial cells were significantly increased in RS3 group compared to the constipated cellulose group. Supplementation of resistant starch significantly elevated the ratio of HDL-cholesterol to total cholesterol and significantly lowered plasma atherogenic index compared with cellulose or pectin supplementation in constipated rats. The results of the present study demonstrated that resistant starch supplementation may help in improving the large bowel environment by stimulation of bifidobacterial proliferation, reduction of pH and inflammation factor and by increased production of mucus. It has also been found that an additional health benefit is improvement in lipid levels of serum.

Ichthyotoxic Cochlodinium polykrikoides red tides offshore in the South Sea, Korea in 2014: I. Temporal variations in three-dimensional distributions of red-tide organisms and environmental factors

  • Jeong, Hae Jin;Lim, An Suk;Lee, Kitack;Lee, Moo Joon;Seong, Kyeong Ah;Kang, Nam Seon;Jang, Se Hyeon;Lee, Kyung Ha;Lee, Sung Yeon;Kim, Mi Ok;Kim, Ji Hye;Kwon, Ji Eun;Kang, Hee Chang;Kim, Jae Seong;Yih, Wonho;Shin, Kyoungsoon;Jang, Poong Kook;Ryu, Joo-Hyung;Kim, Sung Young;Park, Jae Yeon;Kim, Kwang Young
    • ALGAE
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    • v.32 no.2
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    • pp.101-130
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    • 2017
  • The ichthyotoxic Cochlodinium polykrikoides red tides have caused great economic losses in the aquaculture industry in the waters of Korea and other countries. Predicting outbreak of C. polykrikoides red tides 1-2 weeks in advance is a critical step in minimizing losses. In the South Sea of Korea, large C. polykrikoides red tide patches have often been recorded offshore and transported to nearshore waters. To explore the processes of offshore C. polykrikoides red tides, temporal variations in 3-dimensional (3-D) distributions of red tide organisms and environmental parameters were investigated by analyzing 4,432 water samples collected from 2-5 depths of 60 stations in the South Sea, Korea 16 times from May to Nov, 2014. In the study area, the vegetative cells of C. polykrikoides were found as early as May 7, but C. polykrikoides red tide patches were observed from Aug 21 until Oct 9. Cochlodinium red tides occurred in both inner and outer stations. Prior to the occurrence of large C. polykrikoides red tides, the phototrophic dinoflagellates Prorocentrum donghaiense (Jun 12 to Jul 11), Ceratium furca (Jul 11 to Aug 21), and Alexandrium fraterculus (Aug 21) formed red tides in sequence, and diatom red tides formed 2-3 times without a certain distinct pattern. The temperature for the optimal growth of these four red tide dinoflagellates is known to be similar. Thus, the sequence of the maximum growth rates of P. donghaiense > C. furca > A. fraterculus > C. polykrikoides may be partially responsible for this sequence of red tides in the inner stations following high nutrients input in the surface waters because of heavy rains. Furthermore, Cochlodinium red tides formed and persisted at the outer stations when $NO_3$ concentrations of the surface waters were < $2{\mu}M$ and thermocline depths were >20 m with the retreat of deep cold waters, and the abundance of the competing red-tide species was relatively low. The sequence of the maximum swimming speeds and thus potential reachable depths of C. polykrikoides > A. fraterculus > C. furca > P. donghaiense may be responsible for the large C. polykrikoides red tides after the small blooms of the other dinoflagellates. Thus, C. polykrikoides is likely to outgrow over the competitors at the outer stations by descending to depths >20 m and taking nutrients up from deep cold waters. Thus, to predict the process of Cochlodinium red tides in the study area, temporal variations in 3-D distributions of red tide organisms and environmental parameters showing major nutrient sources, formation and depth of thermoclines, intrusion and retreat of deep cold waters, and the abundance of competing red tide species should be well understood.

A Study of Genetic Polymonhisms of HLA-class I and II Genes Using Polymerase Chain Reaction (중합효소연쇄반응을 이용한 HLA-class I, II 유전자군의 유전적 다형성에 관한 연구)

  • Kyung-Ok Lee
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.11-25
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    • 1998
  • The HLA genes located in the short arm of chromosome 6 specify heterodimeric glycoproteins involved in the regulation of the immune response. Recently, in the elucidation of HLA polymorphism, serological and cellular typing methods have been replaced by DNA typing using polymerase chain reaction (PCR). The purpose of this study was to establish the HLA DNA typing methods and determine gene frequencies of HLA molecules in Koreans. PCR-SSP (sequence specific primers) and PCR-RFLP (restriction fragment length polymorphism) techniques were used for the analysis of HLA-A, -B, -C, DRBl genes and HLA-DQAl, DQBl, DPBl genes, respectively. The results of B-lymphoblastoid cells used for control experiment were consistent with the previous data identified in the 11th International Histocompatibility Workshop. Seventeen, 23, 16, 8, 16, 13 and 37 types of HLA-A, B, C, DQAl, DQBl, DPBl and DRBl alleles were found, respectively, in a total of unrelated 120 Korean individuals. The most frequent HLA alleles were $A^*$02 (27.0%), B$^*$40 (17.6%), Cw$^*$01 (19.2%), DQAl$^*$0301 (32.1%), DQBl$^*$0303 (12.9%), DPBl$^*$0501 (31.3%) and DRBl$^*$1501 (9.2%) among Koreans. This study shows that DNA typing method using PCR technique is a relatively simple, fast and practical tool for the determination of the HLA-class I and II genes. Moreover, the data of HLA gene frequencies could be useful for the Korean database before clinical applications, including organ and unrelated bone marrow transplantation, anthropological study, disease association and individual identification.

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Ionic currents elicited by the hypotonic solution in hamster eggs (저장성 용액에 노출된 햄스터 난자에 관찰되는 이온전류의 변화)

  • Choi, Won-yeong;Kim, Yang-mi;Haan, Jae-hee;Huh, Il-oh;Park, Choon-ok;Hong, Seong-geun;Pyu, Pan-dong;Kim, Jong-shu
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.305-312
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    • 1996
  • Cell volume regulatory mechanisms are usually disclosed by exposure of cell to anisotonic media. If a cell is suddenly exposed to hypotonic media, it swells initially like an osmometer but within minutes regains its original cell volume. This behavior has been labelled as regulatory cell volume decrease(RVD). RVD is believed to result from the loss of permeable ions through the membrane. In this study, we examined hypotonically induced changes in the membrance currents involved in RVD by using whole cell voltage clamp technique in the unfertilized hamster egg. At -40mV of the holding potential, the stationary current was maintained in the hamster egg exposed to isotonic solution composed of, mainly, 115mM NaCl and 40mM mannitol. Hypotonic solution was prepared by removing mannitol. Therefore, the concentrations of $Na^+$ and $Cl^-$ in this hypotonic media were the same as those in the isotonic solution. Following 30 to 60 sec after applying the hypotonic media to the egg, the inward current was evoked. This inward current was eliminated by $100{\mu}M$ 4-acetamido-4'-isothiocyanostil-bene-2,2'-disulfonic acid(SITS), an anion channel blocker, leaving the small outward current component. Further addition of 2mM $Ba^{2+}$, a broad $K^+$ channel blocker, completely abolished the small outward current left even in the presence of SITS during hypotonic stress. These results suggest that $K^+$ and $Cl^-$ move out of cells, resulting in RVD. To test the involvement of $Na^+$ in RVD, 20mM Na-isethionate was substituted for mannitol in isotonic media(135mM $Na^+$) and Na-isethionate (20mM) was freed the hypotonic solution. Only $Cl^-$ concentration in both isotonic and hypotonic media was kept constant at 115mM, whereas concentration of $Na^+$ was lowered in hypotonic solution to 115mM from 135mM in isotonic solution. Hypotonic medium induced the outward current in the egg equilibrated isotonically. This current was reduced by $100{\mu}M$ SITS but was augmented by 2 mM $Ba^{2+}$. In terms of RVD, these results imply that $Cl^-$ efflux is coupled with $K^+$, maybe for electroneutrality during hypotonic stress and/or with $Na^+$ via unknown transport mechanism(s). From the overall results, the hypotonic stress facilitates the movement of $Cl^-$ and $K^+$ out of the hamster egg to regain cellular volume with electroneutrality. If there exist a difference in $[Na^+]_0$ between isotonic and hypotonic solution, another transport mechanism concerned with $Na^+$ may, at least partly, participate in regulatory volume decrease.

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The Effects of Microcurrent Stimulation on the Astrocytes Proliferation at Injured Brain of Rabbit (극저전류자극이 손상된 토끼 뇌의 별아교세포 증식에 미치는 효과)

  • Kim, Ji-Sung;Min, Kyoung-Ok
    • Journal of Korean Physical Therapy Science
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    • v.9 no.3
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    • pp.107-119
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    • 2002
  • Astrocyte, which shares the greatest part of the brain (about 25%), is a land of glial cell that composes the central nervous system along with microglia, ependymal cell and oligodendroglia. It has 7-9nm of fibers in its cytoplasma, which are composed of glial fibrillary acidic protein (GFAP) and vimentin. As for the functions of the astrocyte, it has, so far, been supposed that the astrocyte will play a cytoskeletal role in maintaining the structure of the cerebrum, play a role as a blood-brain barrier so that it can induce migration of the neuron in its development and substances in the blood cannot go into the nervous tissue, and a role of immunology and phagocytosis. However, it was revealed today that it will be a role in preventing expansion of injury by attaching itself to the connective tissue such as the vessel and the pia mater when the nervous tissue or the arachnoid is injured. Microcurrent stimulation can control current, on the basis of A unit. That is, with such devices using it, it is possible to sense, from the outside, the injured current(wound current) of the lesion and to change it into the normal current, thereby promoting the restoration of the cells. In order to examine the effects of microcurrent stimulation on the injured astrocytes in the rabbits, this study was conducted with 24 New Zealand White Rabbit as its subjects, which were divided into 8 animals of the experiment group and 16 animals of the control group. After the animals in the experiment group were fixed to the stereotaxic apparatus, their hair was removed and their premotor area(association area) perforated by the micro-drill for skull-perforation with the depth of 8mm from the scalp. In one week after the injury, 4 animals in the control group and 8 animals in the experiment group were sacrificed and examined with immunohistochemical method. And in three weeks, the remaining 4 animals in the control group and 8 animals in the experiment group were also sacrificed and examined with the same way. The conclusion has been drawn as follows : In the control group sacrificed in one week after the injury, the astrocytes somewhat increased, compared with the normal animals, and in the group sacrificed in three weeks after the injury, they increased more (p < 0.05). The experiment group A in one week showed a little increase, but there was no significant differences, but the experiment group in three weeks showed more increase, compared with the experiment group in one week (p < 0.05). The experiment group B in one week showed more increase than the control group or the experiment group A, and the experiment group in three weeks showed more increase than the experiment group in one week (p < 0.05). Among the astrocytes, fibrous astrocytes were mostly observed, increasing as they are close to the lesion, and decreasing as they are remote from it. The findings show that microcurrent can cause the astrocytes to proliferate and that it will be more effective to stimulate the cervical part somewhat remote from the lesion rather than to directly stimulate the part of the lesion. Thus, microcurrent stimulation can be one of the methods that can activate the reaction of astrocytes, which is one of the mechanism for treating cerebral injury with hemorrhage. Therefore, this study will be used as basic research data for promoting restoration of functions in the patient with injury in the central nervous system.

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Effect of Small Dose of Radiation on Induction of Apoptosis in Murine Tumors (마우스 종양에서 저선량 방사선이 Apoptosis의 유도에 미치는 영향)

  • Seong, Jin-Sil;Pyo, Hong-Ryull;Chung, Eun-Ji;Kim, Sung-Hee;Suh, Chang-Ok
    • Radiation Oncology Journal
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    • v.17 no.4
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    • pp.307-313
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    • 1999
  • Purpose : To investigate the Presence of adaptive response by low dose radiation in murine tumors in relation to radiation induced apoptosis as well as related mechanism. Materials and Methods : Syngeneic murine tumors, OCa-I and HCa-l, were given 0.05 Gy pretreatment followed by therapeutic dose of 25 Gy radiation. Induction of apoptosis was analyzed for each treatment group. Regulating molecules of apoptosis, p53, Bcl-2, Bax, Bcl-X, were also analyzed by Western blotting. Results : In 0.05 Gy pretreatment group of OCa-I, 25 Gy-induced apoptosis per 1000 cells was 229, which was estimated at $30\%$ lower level than the expected (p<0.05). In contrast, this reduction in radiation induced apoptosis was not seen in HCa-l. In the expression of apoptosis regulating molecules, p53 increased in both tumors in response to radiation. Bcl-2 and Bax did not show significant change in both tumors however, the expression of Bcl-2 surpassed that of Bax in 0.05 Gy pretreatment group of OCa-l. Bcl-X was not expressed in OCa-l. In HCa-l, Bcl-X showed increased expression even with 0.05 Gy. Conclusion : Adaptive response by low dose radiation Is shown in one murine tumor, OCa-l, in relation to radiation induced apoptosis. Apoptosis regulating molecules including Bcl-2/Bax and Bcl-X, appear to related. This study shows an evidence that adaptive response is present, but not a generalized phenomenon in vivo.

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