• Title/Summary/Keyword: f/2 media

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Expression of Cell Cycle Related Genes in HL60 Cells Undergoing Apoptosis by X-irradiation (HL60 세포주에서 방사선 조사에 의한 Apoptosis와 세포 주기 관련 유전자의 발현 변화)

  • Kim, Jin-Hee;Park, In-Kyu
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.377-388
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    • 1998
  • Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.

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In Vitro Development of Mouse Parthenogenetic Embryos: Effect of Temperature before Oocyte Activation

  • Roh Sangho;Won Cheolhee;Min Byung-Moo
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.117-120
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    • 2005
  • This study was conducted to establish the optimal temperature condition before oocyte activation in B6m F1 mouse. In experiment 1, two embryo culture media (CZB vs KSOM) were evaluated for the development of activated mouse oocytes. Parthenogenetic embryos cultured in KSOM showed better blastocyst development than ones cultured in CZB $(56.2\%\;vs\;81.0\%\;p<0.01)$. Two-hour of pre-incubation before activation significantly reduced the number of hatched blastocysts in KSOM $(22.0\%\;versus\;8.8\%\;p<0.05)$. In experiment 2, recovered oocytes were pre-incubated at different temperature conditions before activation. The experimental groups were divided by 5 as follows. Group A: pre-incubation for 120 min at $37^{\circ}C$, Group B: pre-incubation at $37^{\circ}C$ for 90 min then at $25^{\circ}C$ for 30 min, Group C: pre-incubation at $37^{\circ}C$ for 60 min then at $25^{\circ}C$for 60 min, Group D: pre-incubation at $37^{\circ}C$ for 30 min then at $25^{\circ}C$ for 90 min, and Group E: pre-incubation at $25^{\circ}C$ for 120 min before activation. Group A $(67.6\%)$ and B $(66.7\%)$ showed better development to the blastocyst stage than other groups $(Group\;C:\;50.0\%\;Group \;D:\;49.2\%\;Group\;E:\;33.3\%,\;p<0.05)$. The present study indicates that the temperature before activation affects the development of B6D2 F1 mouse parthenogenetic oocytes and exposure to room temperature should be limited to 30-min when the oocytes are left in HEPES-buffered medium for micromanipulation.

Induction of Plants through Anther Culture of Radish (무 약배양에 의한 식물체 유기)

  • Yun, Yeo-Jung;An, Chun-Hui;Yun, Hwa-Mo
    • The Journal of Natural Sciences
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    • v.6 no.1
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    • pp.71-78
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    • 1993
  • Five $F_1$ hybrids of radish(Raphanus sativus L.) were used in the study for induction of microspore derived embryos. Anthers from the mid-uninucleate to early bicellular stage were inoculated on the modified B5 medium and modified Nitch-Nitch medium supplemented with several growth regulators. The efficiency of anther culture was dspendent on the genotype of donor plants and we obtained various culture efficiency from different genotypes. Induction of embryos from microspore was best result on Nitsch-Nitsch media supplemented with 0.1mg/l NAA and 0.05mg/l BAP. Heat treatments of anthers at $35^{\circ}C$-2days and combined with pretreatment of $4^{\circ}C$ for 2, 8, 12 and 16days . Among the treatments, $35^{\circ}C$-2 days treatment combined with $4^{\circ}C$-2days pretreatment treatment were the most effective in developing embryos from microspores.

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Identification of Antagonistic Streptomyces Species on Phytophthora nicotianae var. parasitica and Fusarium oxysporum sporum f. sg. vasinfectum Causing Sesame Wilt and Blight (참깨 역병(Phytophthora nicotianae var. parasitica) 및 시들음병(Fusarium oxysporum f. sg. vasinfectum)에 길항적인 Streptomyces spp.의 분류 동정)

  • Chung, Bong-Koo;Ser, Sang-Oh
    • The Korean Journal of Mycology
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    • v.20 no.1
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    • pp.65-71
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    • 1992
  • The two isolates of Streptomyces antagonistic to Phytophthora nicotianae var. parasitica and Fusarium oxysporum f. sp. vasinfectum were identified as based on the morphological, cultural and physiological characteristics on various culture media. Spore chains of St-11 isolate was rectus-flexibilis(RF), whereas the other isolate, St-20, was shown rectinaculum-apertum(RA). Spore surface of St-11 isolate was smooth, while St-20 was spiny. Aerial mycelia of the two isolates were all gray color and growing conditions on media were good as a whole. Any soluble pigment was not shown in cultivation of the two isolates. Stoll isolate showed negative response on starch hydrolysis and gelation liquefaction, whereas St-20 isolate was positive on starch hydrolysis and a negative on gelatin reaction. Stoll isolate was identified as Streptomyces bikiniensis and St-20 Streptomyces echinoruber, respectively.

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The Effects of Vero Cell Co-culture on Mouse Embryo Development (Vero Cell과의 공동배양이 체외에서 생쥐 배아발생에 미치는 영향)

  • Lee, Yoon;Park, June-Hong;Kang, He-Na;Kim, Yong-Bong;Lee, Eung-Soo;Park, Sung-Kwan
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.2
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    • pp.233-239
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    • 1997
  • Embryos of most mammalian species grown in vitro would undergo developmental arrest at the approximate time of genomic activation. Stage-specific cell block and the resulting rapid loss of embryo viability in conventional culture media have limited the duration for which embryos may be cultured prior to transfer. As a result, embryos are usually transferred to the uterus at the 4-to 8-cell stage to avoid the loss of viability associated with long-term in vitro culture. Early transfer has led to asynchrony of the endometrium-trophectoderm interaction at the time of implantation and a resultant reduction in the rate of implantation. To overcome these problems, a variety of co-culture systems has been devised in which embryos can develop for a longer period prior to embryo transfer. Vero cells, derived from African green monkey kidney, share a common embryologic origin with cells from the genital tract. In addition, they are potentially safe to use, since they are highly controlled for viruses and other contaminants. Therefore, co-culture using Vero cells has been widely utilized to enhance embryo viability and development, although not without controversies. We thus designed a series of experiments to demonstrate whether Vero cells do indeed enhance mouse embryo development as well as to compare the efficacy of co-culturing mouse 1-cell embryos on Vero cell monolayer in both Ham's F-10 and human tubal fluid (HTF) culture media. 1-cell stage ICR mouse embryos were cultured either in the presence of Vero cells (Group A) or in conventional culture medium alone (Group B). In Ham's F-10 significantly more 3-to-8cell embryos developed in group A than group B (59.8 versus 10.0%; p<0.01). In contrast, there was no significant difference in embryonic development both group A and group B in HTF. However, significant differences were noted only in later embryonic stage (13 and 0%; p<0.05 of group A and B respectively, hatching or hatched). In Ham's F-10, we also could observe the beneficial effect of Vero cell on hatching process (70.7 and 42.1%; p<0.05 of group A and group B respectively).

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PFC Ultrasonic Decontamination Efficiency on the Various Types of Metal Specimens (금속 시편 형태에 따른 PEC 초음파 제염 성능)

  • Won Hui-Jun;Kim Gye-Nam;Jung Chung-Hun;Park Jin-Ho;Oh Won-Zin
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.3 no.4
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    • pp.293-300
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    • 2005
  • Ultrasonic decontamination of the type 304 stainless steel specimen loosely contaminated with $Eu_2O_3$ powders was investigated. Decontamination factors (DFs) by the three kinds of ultrasonic media such as water, pure PFC (Pefluorocarbon, $C_7F_{16}$) and a mixed solution of $99.9\;vol\%\;PFC\;and\;0.1\;vol\%$ anionic surfactant were determined. The determined DF values were 20, 50 and 200, respectively. This significant difference in the decontamination factors for the different decontamination solution was well explained by the surface tension of the media as well as the interaction between the positively charged surface of $Eu_2O_3$ powders and the anionic surfactant. Ultrasonic decontamination behavior of the loosely contaminated metal specimens such as plate, pipe, welding specimen and crevice specimen in the mixed solution of PFC and anionic surfactant was also investigated. The contaminants were completely removed for the tested specimens except for the longest specimen. For 6-cm long pipe specimen, however, $98.5\%$ of the contaminants were removed.

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Expression of the Promoter for the Maltogenic Amylase Gene in Bacillus subtilis 168

  • Kim Do-Yeon;Cha Choon-Hwan;Oh Wan-Seok;Yoon Young-Jun;Kim Jung-Wan
    • Journal of Microbiology
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    • v.42 no.4
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    • pp.319-327
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    • 2004
  • An additional amylase, besides the typical $\alpha-amylase,$ was detected for the first time in the cytoplasm of B. subtilis SUH4-2, an isolate from Korean soil. The corresponding gene (bbmA) encoded a malto­genic amylase (MAase) and its sequence was almost identical to the yvdF gene of B. subtilis 168, whose function was unknown. Southern blot analysis using bbmA as the probe indicated that this gene was ubiquitous among various B. subtilis strains. In an effort to understand the physiological function of the bbmA gene in B. subtilis, the expression pattern of the gene was monitored by measuring the $\beta-galactosidase$ activity produced from the bbmA promoter fused to the amino terminus of the lacZ struc­tural gene, which was then integrated into the amyE locus on the B. subtilis 168 chromosome. The pro­moter was induced during the mid-log phase and fully expressed at the early stationary phase in defined media containing $\beta--cyclodextrin\;(\beta-CD),$ maltose, or starch. On the other hand, it was kept repressed in the presence of glucose, fructose, sucrose, or glycerol, suggesting that catabolite repression might be involved in the expression of the gene. Production of the $\beta-CD$ hydrolyzing activity was impaired by the spo0A mutation in B. subtilis 168, indicating the involvement of an additional regu­latory system exerting control on the promoter. Inactivation of yvdF resulted in a significant decrease of the $\beta-CD$ hydrolyzing activity, if not all. This result implied the presence of an additional enzyme(s) that is capable of hydrolyzing $\beta-CD$ in B. subtilis 168. Based on the results, MAase encoded by bbmA is likely to be involved in maltose and $\beta-CD$ utilization when other sugars, which are readily usable as an energy source, are not available during the stationary phase.

The physiological characteristics of Fomers fomentarius. (말굽버섯의 생리적 특성)

  • 채정기;황태익;서승현;김현석;장경수;문형률;주명옥;박병인
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.60-60
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    • 2002
  • This study was excuted to decide the physiological characteristics of Fomers fomentarius. F. fomentarius was tested to select pertinent substract, temperature and pH range for the growth. Mycelial growth of F. fomentarius was mostly supported on MYPA among other tested synthetic or semi-synthetic media. The temperature range for pertinent mycelial growth was about 22~32$^{\circ}C$ and mostly stimulated at $25^{\circ}C$. And the pertinent pH range of MYPA was 5.0~6.0. The required carbon and nitrogen source for mycelial growth of F. fomentarius was tested. The mycelial growth was mostly stimulated by soluble starch at content. The carbon sources for pertinent mycelial growth was glucose or starch. And the nitrogen source for pertinent mycelial growth was yeast

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Degradation Characteristics of Wood Cellulose by Ruminal Cellulolytic Anaerobic Bacterium Ruminococcus albus F-40 (혐기성 세균 Ruminococcus albus F-40에 의한 목재 cellulose의 분해특성)

  • Kim, Yoon-Soo;Wi, Seung-Gon;Myung, Kyu-Ho
    • Journal of the Korean Wood Science and Technology
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    • v.25 no.3
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    • pp.83-95
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    • 1997
  • The degradation mode of lignocellulose by anaerobic ruminal cellulolytic bacterium Ruminococcus albus F-40 was investigated. Birchwood holocellulose and filter paper were incubated as the sole carbohydrate sources with using the Hungate techniques. After 2 or 4 days of incubation, samples were employed for chemical and electron microscopic evaluations. The degradation rate of cellulosic substrates and the adhesion rate of bacteria to the substrates increased proportionally with the decrease of relative crystallinity of cellulose, indicating the preferential breakdown of amorphous cellulose, by this bacterium. X-ray diffraction analyses and polarized light microscopy showed, however, that crystalline cellulose was also degraded by R. albus. FT-IR spectra indicated that not only cellulose but hemicellulose was also degraded by this bacterium. Electron microscopic investigations showed the protuberant structures on the surface of R. albus. These structures were much more significant when bacterial cells were grown in the media containing insoluble substrates, such as cellulose, indicating clearly that bacterial protuberant structures were induced by the substrates. Protuberant structures extended from the bacterial cells adhered tightly to the substrates and numerous vesicles covered the surface of cellulosic substrates affected. Cellulosome-like structures were distributed on the cellulose matrix. Electron microscopic works showed that diverse surface organells of R. albus were involved in the degradation of cellulosic materials. SEM examinations showed the breakdown of cellulose by R. albus was proceeded by severeal routes : short fiber formation, defibrillation and destrafication of cellulose microfibril.

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The Effect of PET/CT Images on SUV with the Correction of CT Image by Using Contrast Media (PET/CT 영상에서 조영제를 이용한 CT 영상의 보정(Correction)에 따른 표준화섭취계수(SUV)의 영향)

  • Ahn, Sha-Ron;Park, Hoon-Hee;Park, Min-Soo;Lee, Seung-Jae;Oh, Shin-Hyun;Lim, Han-Sang;Kim, Jae-Sam;Lee, Chang-Ho
    • The Korean Journal of Nuclear Medicine Technology
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    • v.13 no.1
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    • pp.77-81
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    • 2009
  • Purpose: The PET of the PET/CT (Positron Emission Tomography/Computed Tomography) quantitatively shows the biological and chemical information of the body, but has limitation of presenting the clear anatomic structure. Thus combining the PET with CT, it is not only possible to offer the higher resolution but also effectively shorten the scanning time and reduce the noises by using CT data in attenuation correction. And because, at the CT scanning, the contrast media makes it easy to determine a exact range of the lesion and distinguish the normal organs, there is a certain increase in the use of it. However, in the case of using the contrast media, it affects semi-quantitative measures of the PET/CT images. In this study, therefore, we will be to establish the reliability of the SUV (Standardized Uptake Value) with CT data correction so that it can help more accurate diagnosis. Materials and Methods: In this experiment, a total of 30 people are targeted - age range: from 27 to 72, average age : 49.6 - and DSTe (General Electric Healthcare, Milwaukee, MI, USA) is used for equipment. $^{18}F$- FDG 370~555 MBq is injected into the subjects depending on their weight and, after about 60 minutes of their stable position, a whole-body scan is taken. The CT scan is set to 140 kV and 210 mA, and the injected amount of the contrast media is 2 cc per 1 kg of the patients' weight. With the raw data from the scan, we obtain a image showing the effect of the contrast media through the attenuation correction by both of the corrected and uncorrected CT data. Then we mark out ROI (Region of Interest) in each area to measure SUV and analyze the difference. Results: According to the analysis, the SUV is decreased in the liver and heart which have more bloodstream than the others, because of the contrast media correction. On the other hand, there is no difference in the lungs. Conclusions: Whereas the CT scan images with the contrast media from the PET/CT increase the contrast of the targeted region for the test so that it can improve efficiency of diagnosis, there occurred an increase of SUV, a semi-quantitative analytical method. In this research, we measure the variation of SUV through the correction of the influence of contrast media and compare the differences. As we revise the SUV which is increasing in the image with attenuation correction by using contrast media, we can expect anatomical images of high-resolution. Furthermore, it is considered that through this trusted semi-quantitative method, it will definitely enhance the diagnostic value.

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