• Title/Summary/Keyword: In vitro survival

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Increased Catalase Activity by All-trans Retinoic Acid and Its Effect on Radiosensitivity in Rat Glioma Cells (백서 교종 세포에서 레티노인산에 의한 카탈라제의 활성 증가가 방사선감수성에 미치는 효과)

  • Jin, Hua;Jeon, Ha-Yeun;Kim, Won-Dong;Ahn, Hee-Yul;Yu, Jae-Ran;Park, Woo-Yoon
    • Radiation Oncology Journal
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    • v.23 no.4
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    • pp.211-216
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    • 2005
  • Purpose: It has been reported that all-trans retinoic acid (ATRA) can inhibit glioma growing in vitro. However, clinical trials with ATRA alone in gliomas revealed modest results. ATRA has been shown to increase radiosensitivity in other tumor types, so combining radiation and ATRA would be one of alternatives to increase therapeutic efficacy in malignant gliomas. Thus, we intended to know the role of catalase, which is induced by ATRA, for radiosensitivity if radiation-reduced reactive oxygen species (ROS) is removed by catalase, the effect of radiation will be reduced. Materials and Methods: A rat glioma cell line (36B10) was used for this study. The change of catalase activity and radiosensitivity by ATRA, with or without 3-amino-1, 2, 4-triazole (ATZ), a chemical inhibitor of catalase were measured. Catalase activity was measured by the decomposition of $H_2O_2$ spectrophotometrically Radiosensitivity was measured with clonogenic assay. Also ROS was measured using a 2, 7-dichlorofluorescein diacetate spectrophotometrically. Results: When 36B10 cells were exposed to 10, 25 and $50{\mu}M$ of ATRA for 48 h, the expression of catalase activity were increased with increasing concentration and incubation time of ATRA. Catalase activity was decreased with increasing the concentration of AT (1, $10{\mu}M$) dose-dependently. ROS was increased with ATRA and it was augmented with the combination of ATRA and radiation. ATZ decreased ROS production and increased cell survival in combination of ATRA and radiation despite the reduction of catalase. Conclusion: The increase of ROS is one of the reasons for the increased radiosensitivity in combination with ATRA. The catalase that is induced by ATRA doesn't decrease ROS production and radiosensitivity.

Anti-Stress Effect of Punica granatum L. Extract against Sleep Deprivation-Induced Impairment (석류 열수 추출물의 수면박탈을 유도한 Rat 모델에서의 항스트레스 효과)

  • Na, Ju-Ryun;Kim, Sunoh;Jo, Ara;Bae, Donghyuck;Oh, Kyo-Nyeo;Kim, Yong Jae;Lee, Yoo-Hyun;Jun, Woojin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.11
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    • pp.1533-1543
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    • 2016
  • The anti-stress effects of Punica granatum L. (family Lythraceae, PG) on $H_2O_2$/corticosterone (CORT)-induced stress in cells and sleep-deprived rats were investigated. The PG extract showed neuroprotective effects in SH-SY5Y cells against $H_2O_2$/CORT-induced stress. Sleep deprivation led to behavioral, hormonal, and biochemical alterations in the animal model. The effects of P. granatum on physiological, behavioral, and biochemical parameters aggravated by sleep deprivation were investigated. Sleep deprivation impaired physiological (survival, body weight, and drowsiness scores) and behavioral (rotarod, passive avoidance, hot hyperalgesia, and Y maze) parameters as well as biochemical factors (cortisol, serotonin, dopamine, testosterone, and growth factor I contents in serum). These parameters were significantly recovered by PG extract in a concentration-dependent manner. The PG extract also enhanced catalase, superoxide dismutase, and non-enzymatic antioxidative activities such as glutathione compared to sleep-deprived rats. On the basis of these results, our findings suggest that Punica granatum prevents impairment of body functions induced by sleep deprivation and related oxidative damage.

Biochemical Assessment of Deer Velvet Antler Extract and its Cytotoxic Effect including Acute Oral Toxicity using an ICR Mice Model (ICR 마우스 모델을 이용한 녹용 추출물의 생화학적 평가 및 급성 경구 독성을 포함한 세포 독성 효과)

  • Ramakrishna Chilakala;Hyeon Jeong Moon;Hwan Lee;Dong-Sung Lee;Sun Hee Cheong
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.430-441
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    • 2023
  • Velvet antler is widely used as a traditional medicine, and numerous studies have demonstrated its tremendous nutritional and medicinal values including immunity-enhancing effects. This study aimed to investigate different deer velvet extracts (Sample 1: raw extract, Sample 2: dried extract, and Sample 3: freeze-dried extract) for proximate composition, uronic acid, sulfated glycosaminoglycan, sialic acid, collagen levels, and chemical components using ultra-performance liquid chromatography-quadrupole-time-of-light mass spectrometry. In addition, we evaluated the cytotoxic effect of the deer velvet extracts on BV2 microglia, HT22 hippocampal cells, HaCaT keratinocytes, and RAW264.7 macrophages using the cell viability MTT assay. Furthermore, we evaluated acute toxicity of the deer velvet extracts at different doses (0, 500, 1000, and 2000 mg/kg) administered orally to both male and female ICR mice for 14 d (five mice per group). After treatment, we evaluated general toxicity, survival rate, body weight changes, mortality, clinical signs, and necropsy findings in the experimental mice based on OECD guidelines. The results suggested that in vitro treatment with the evaluated extracts had no cytotoxic effect in HaCaT keratinocytes cells, whereas Sample-2 had a cytotoxic effect at 500 and 1000 ㎍/mL on HT22 hippocampal cells and RAW264.7 macrophages. Sample 3 was also cytotoxic at concentrations of 500 and 1000 ㎍/mL to RAW264.7 and BV2 microglial cells. However, the mice treated in vivo with the velvet extracts at doses of 500-2000 mg/kg BW showed no clinical signs, mortality, or necropsy findings, indicating that the LD50 is higher than this dosage. These findings indicate that there were no toxicological abnormalities connected with the deer velvet extract treatment in mice. However, further human and animal studies are needed before sufficient safety information is available to justify its use in humans.

Evaluation of Sprouted Barley as a Nutritive Feed Additive for Protaetia brevitarsis and Its Antibacterial Action against Serratia marcescens (흰점박이꽃무지 사료첨가제로서 새싹보리의 곤충병원성 세균에 대한 항균 효과에 관한 연구)

  • Song, Myung Ha;Kim, Nang-Hee;Park, Kwan-Ho;Kim, Eunsun;Kim, Yongsoon
    • Journal of Life Science
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    • v.31 no.5
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    • pp.475-480
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    • 2021
  • Interest in edible insects such as Protaetia brevitarsis has increased rapidly, and several insect producers use these insects in industrialized mass production. However, mass rearing of insects can cause insect diseases. Sprouted barley is a valuable source of nutrients and has antioxidant, antimicrobial, anti-inflammatory, and anti-cancer effects. This study was conducted to investigate the effect of sprouted barley as a feed additive for producing healthy P. brevitarsis larvae. P. brevitarsis larvae were fed feeds with or without sprouted barley, and their body weight and larval period wewe checked weekly. To confirm the antibacterial effects of sprouted barley, in vitro bioassays were performed by counting Serratia marcescens colonies, and in vivo bioassays were performed by determining the survival rate and body weights of the S. marcescens-infected larvae. Larvae fed different feeds were analyzed for their nutrient compositions (i.e., such as proximate composition, minerals, amino acids, and heavy metals). Larvae fed 5% and 10% sprouted barley had maximum weight increases of 19.2% and 23.1%, respectively. Both treatment groups had significantly shorter larval periods than those of the control group. Sprouted barley markedly inhibited the growth of entomopathogenic S. marcescens. Furthermore, larvae fed sprouted barley exhibited higher Cu, Zn, and K levels. Seventeen amino acids were present in larvae fed sprouted barley, of which, tyrosine and glutamic acid were predominant. No heavy metals were detected in any of the investigated groups. Therefore, sprouted barley may be a suitable feed additive for producing high-quality P. brevitarsis larvae.

Developmental Capacity of Bovine Follicular Oocytes after Ultra-Rapid Freezing by electron Microscope Grid II.Cryopreservation of In Vitro Matured Bovine Oocytes (Electron Microscopic Grid를 이용한 초급속 동결이 소 난포란의 발달능에 미치는 영향. II. 체외 성숙된 소 미수정란의 동결에 관한 연구)

  • 김은영;김남형;이봉경;윤산현;박세필;정길생;임진호
    • Korean Journal of Animal Reproduction
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    • v.22 no.1
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    • pp.1-9
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    • 1998
  • This study was carried out to confirm whether the developmental capacity of bovine mature oocytes frozen ultra-rapidly using electron microscopic(EM) grids and EFS30 can be obtained, and whether the cryoprotectants and the freezing method used in this study effect detrimentally to the bovine oocytes by indirect immunocytochemistry. As freezing solution, we used EFS30 which consisted of 30% ethylene glycol, 0.5 M sucrose, 18% ficoll and 10% FBS added in D-PBS. The results obtained in this experiment were summarized as follows: When the effects of cryoprotectant and freezing procedure on the microtuble, micrfilament and chromatin morphology of oocytes were evaluated using indirect immunocytochemistry, the results of freezing as well as exposure group were not different with that of the control oocytes. When the fertilization abnormality after ultrarapid freezing of bovine mature oocytes was examined by Hoechst staining, the rates of total penetration(96.7, 9.0%), normal two pronuclei formation(74.6, 68.9%) and mean number of sperm / oocyte(1.50, 1.44) were not different between control and freezing group. In addition, when the developmental capacity of frozen-thawed of oocytes(85.5%) was survived, 74.5% of them were cleaved and 31.4% of cleaved embryos were developed to blastocyst. These data were similar to those of the control(76.0%, 34.6%) and exposure(74.5%, 33.0%) except survival rates. Also, when the total cell number of blastocysts produced from the each treatment at day after IVF was examined by hoechst staining, there were not different among groups. There results demonstrate that developmental capacity of frozen-thawed bovine mature oocytes can be successfully obtained by ultra-rapid freezing method using EM grid and EFS30 solution.

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Physicochemical properties and microencapsulation process of rice fermented with Bacillus subtilis CBD2 (Bacillus Subtilis CBD2로 배양된 백미 발효물의 미세캡슐 제조 및 물리화학적 특성)

  • Lee, Dae-Hoon;Park, Hye-Mi;Hong, Joo-Heon
    • Food Science and Preservation
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    • v.22 no.2
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    • pp.225-231
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    • 2015
  • This study was conducted to examine the physicochemical properties and micro-encapsulation process of rice fermented with Bacillus subtilis CBD2. The viable bacterial cell, pH, and amylase activity of the rice liquid culture were 7.61 log CFU/mL, pH 5.08 and 159.43 units/mL, respectively. The micro-encapsulated rice liquid culture was manufactured via spray drying with different forming agents: i.e., alginic acid 1.0% and chitosan 0.3%, 0.5%, and 1.0%. The moisture contents of the spray-dried powders were approximately 2.90~3.68%. The color of the L and a value decreased whereas that of the b and ${\Delta}E$ value increased. The particle size and outer topology of the spray-dried rice liquid culture were $48.13{\sim}68.48{\mu}m$ and globular, respectively. The water absorption index of the spray-dried powder (2.40~2.65) was lower than that of the freeze-dried powder (2.66). The water solubility index of the spray-dried powder (9.17~10.89%) was higher than that of the freeze-dried powder (7.12%). The in vitro dissolution was measured for five hours in pH 1.2 simulated gastric fluid, and pH 6.8 and pH 7.4 simulated intestinal fluids, using a dissolution tester at $37^{\circ}C$ with 50 rpm agitation. The amylase survival in the fermented rice was 85.93% through the spray-drying and it was very effectively controlled.

Establishment of optimal conditions for micropropagation by node culture and multiple shoots formation from sucker explants of thornless Blackberry (Rubus fruticosus L. cv. BB21) (가시없는 블랙베리(Rubus fruticosus L. cv. BB21)의 근맹아를 이용한 다경유도와 절간배양을 통한 식물체 증식조건의 확립)

  • Lee, Kang Seop;Kim, Hyo Jin;Park, Dae Hyun;Oh, Seung Cheol;Cho, Han Jig;Kim, Ee Youb
    • Journal of Plant Biotechnology
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    • v.45 no.2
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    • pp.110-116
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    • 2018
  • This study was conducted to develop a simple, rapid, and reliable method for in vitro propagation of disease-free and true-to-type clones from sucker explants of thornless blackberry (Rubus fruticosus L. ${\times}$ R. parvifolius L.). To induce multiple shoots, the sucker explants were sterilized in 1% NaOCl solution, and then were aseptically cultured on the full and 1/2 MS solid medium supplemented with BAP (0.1, 0.5, 1.0, 2.0 mg/L). After six weeks of culture, the highest frequency (85.4%) of shoot formation from sucker explants was obtained on the full-strength MS medium with 1.0 mg/L BAP. Node explants obtained from multiple shoots were cultured on the various media of full- or half-strength of AD, B5, MS, SH, QL, WPM media, respectively. After 30 days of culture, plant growth was good on the half-AD, half-QL medium. After 90 days of culture, plant growth was good on the full MS and full SH medium. The survival rate of the plantlets after transfer to plastic pots containing soil mixture (sand: soil: vermiculite was 1:1:1, vol.) in the greenhouse was 98%. The results indicate that a multiple-shoot procedure can be applied for an efficient mass propagation of Rubus fruticosus L. ${\times}$ R. parvifolius L.

Influence of Temperature on the Egg Production and Hatching of Microcotyle sebastis (Monogenea : Microcotylidae), Parasitic on Rockfish, Sebastes schlegeli (수온이 조피볼락에 기생하는 아가미흡충(Microcotyle sebastis)의 산란과 부화에 미치는 영향)

  • Kim, Ki-Hong;Choi, Eun-Seok;Cho, Jae-Bum;Hwang, Yoon-Jung;Park, Soo-Il
    • Journal of fish pathology
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    • v.11 no.2
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    • pp.113-117
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    • 1998
  • The influence of temperature on the rate of egg production and embryonic development of Microcotyle sebastis was investigated to determine the precise time of a second treatment. The survival time of the adults of M. sebastis was inversely proportional to temperature. The number of laid eggs per each replicate during the first 24 h was $39.3{\pm}4.0$ at $10^{\circ}C$, $62.7{\pm}14.2$ at $15^{\circ}C$, $101.0{\pm}5.6$ at $20^{\circ}C$ and $89.0{\pm}11.0$ at $25^{\circ}C$. The time required for egg hatching of M. sebastis was $31.30{\pm}4.88$, $17.52{\pm}3.24$, $11.59{\pm}3.02$ and $10.76{\pm}3.10$ days at 10, 15, 20 and $25^{\circ}C$, respectively. The regression models of the time required for the beginning and 50% point of hatching according to the different temperatures were as follows; Beginning of hatch: D=58.2000-$4.2067{\times}Temp+0.0867{\times}(Temp)^2$ ($P\leq0.01$), 50% of hatch: D=91.3833-$7.5767{\times}Temp+0.1767{\times}(Temp)^2$ ($P\leq0.01$).

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Correlation Between the Parameters of Radiosensitivity in Human Cancer Cell Lines (인체 암세포주에서 방사선감수성의 지표간의 상호관계)

  • Park, Woo-Yoon;Kim, Won-Dong;Min, Kyung-Soo
    • Radiation Oncology Journal
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    • v.16 no.2
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    • pp.99-106
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    • 1998
  • Purpose : We conducted clonogenic assay using human cancer cell lines (MKN-45, PC-14, Y-79, HeLa) to investigate a correlation between the parameters of radiosensitivity. Materials and Methods : Human cancer cell lines were irradiated with single doses of 1, 2, 3, 5, 7 and 10Gy for the study of radiosensitivity and subrethal damage repair capacity was assessed with two fractions of 5Gy separated with a time interval of 0, 1, 2, 3, 4, 6 and 24 hours. Surviving fraction was assessed with clonogenic assay using $Sperman-H\"{a}rbor$ method and mathematical analysis of survival curves was done with linear-quadratic (LQ) , multitarget-single hit(MS) model and mean inactivation dose$(\v{D})$. Results : Surviving fractions at 2Gy(SF2) were variable among the cell lines, ranged from 0.174 to 0.85 The SF2 of Y-79 was lowest and that of PC-14 was highest(p<0.05, t-test). LQ model analysis showed that the values of $\alpha$ for Y-79, MKN-45, HeLa and PC-14 were 0.603, 0.356, 0.275 and 0.102 respectively, and those of $\beta$ were 0.005, 0.016, 0.025 and 0.027 respectively. Fitting to MS model showed that the values of Do for Y-79. MKN-45, HeLa and PC-14 were 1.59. 1.84. 1.88 and 2.52 respectively, and those of n were 0.97, 1.46, 1.52 and 1 69 respectively. The $\v{D}s$ calculated by Gauss-Laguerre method were 1.62, 2.37, 2,01 and 3.95 respectively So the SF2 was significantly correlated with $\alpha$, Do and $\v{D}$. Their Pearson correlation coefficiencics were -0.953 and 0,993. 0.999 respectively(p<0.05). Sublethal damage repair was saturated around 4 hours and recovery ratios (RR) at plateau phase ranged from 2 to 3.79. But RR was not correlated with SF2, ${\alpha}$, ${\beta}$, Do, $\v{D}$. Conclusion : The intrinsic radiosensitivity was very different among the tested human cell lines. Y-79 was the most sensitive and PC-l4 was the least sensitive. SF2 was well correlated with ${\alpha}$, Do, and $\v{D}$. RR was high for MKN-45 and HeLa but had nothing to do with radiosensitivity parameters. These basic parameters can be used as baseline data for various in vitro radiobiological experiments.

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Experimental Studies on the Antitumor Effects of Jinryungtang Gagambang Extract (진령탕가감방의 항종양효과(抗腫瘍效果)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Jeong, Jun-Tak;Moon, Goo;Moon, Suk-Jae
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.4 no.1
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    • pp.37-53
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    • 1998
  • The sprig of Jinryungtang Gagambang has been used for curing as a traditional medicine without any experimental evidence to support the rational basis for their clinical use. This experiment was carried out to evaluate the possible therapeutic or antitumoral effects of Jinryungtang Gagambang extract against cancer, and to study some mechanisms responsible for its effect. The cytotoxic and antitumor effects were evaluated on human cell liens (A549, hep3B, Caki-1, Sarcoma 180) after exposure to Jinryungtang Gagambang extract using in ILS, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. As a result of exposure to Jinryungtang Gagambang extract, the proliferation of A549, hep3B, Caki-1, good correlations were shown from the results of SRB assay and those of clogenetic assay. 2. The oral administration of Jinryungtang Gagambang extract showed significant effects of increase of MST(mean survival time) and ILS(increased life span) depending on the increasing concentration. 3. Against squamous cell carcinoma induced by MCA, Jinryungtang Gagambang decreased not only the frequency of tumor production but also the number and weight of tumors per tumor bearing mice(TBM). Jinryungtang Gagambang also significantly suppressed the development of 3LL cell-implanted tumors by frequency and their size, and some developed tumors were regressed by the continuous treatment of Jinryungtang Gagambang extract into TBM. 4. Jinryungtang Gagambang extract also increased NK cell activities. According to the above results, it could be suggested that Jinryungtang Gagambang extract has prominent antiutmor effect.

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