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Gene Expression Profiling of Genotoxicity Induced by MNNG in TK6 Cell

  • Suh, Soo-Kyung;Kim, Tae-Gyun;Kim, Hyun-Ju;Koo, Ye-Mo;Lee, Woo-Sun;Jung, Ki-Kyung;Jeong, Youn-Kyoung;Kang, Jin-Seok;Kim, Joo-Hwan;Lee, Eun-Mi;Park, Sue-Nie;Kim, Seung-Hee;Jung, Hai-Kwan
    • Molecular & Cellular Toxicology
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    • v.3 no.2
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    • pp.98-106
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    • 2007
  • Genotoxic stress triggers a variety of biological responses including the transcriptional activation of genes regulating DNA repair, cell survival and cell death. In this study, we investigated to examine gene expression profiles and genotoxic response in TK6 cells treated with DNA damaging agents MNNG (N-methyl-N'-nitrosoguanidine) and hydrogen peroxide $(H_2O_2)$. We extracted total RNA in three independent experiments and hybridized cRNA probes with oligo DNA chip (Applied Biosystems Human Genome Survey Microarray). We analyzed raw signal data with R program and AVADIS software and identified a number of deregulated genes with more than 1.5 log-scale fold change and statistical significancy. We indentified 14 genes including G protein alpha 12 showing deregulation by MNNG. The deregulated genes by MNNG represent the biological pathway regarding MAP kinase signaling pathway. Hydrogen peroxide altered 188 genes including sulfiredoxins. These results show that MNNG and $H_2O_2$ have both uniquely regulated genes that provide the potential to serve as biomarkers of exposure to DNA damaging agents.

Fine Structure of Blue-green Algae, Microcystis aeruginosa Kutzing (남조(藍藻) Microcystis aeruginosa Kutzing의 미세구조(微細構造)에 관(關)한 연구(硏究))

  • Choi, Min-Kyu;Kim, Baik-Ho;Mun, Yeun-Ja;Chung, Yeun-Tai;Lee, Jong-Bin;Wui, In-Sun
    • Applied Microscopy
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    • v.26 no.4
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    • pp.389-399
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    • 1996
  • In order to understand the morphological differences between two different organic loadings by its upstream, and to compare with other algal groups with references, the fine structure of blue-green algae, Microcystis aeruginosa Kitzing, taken from two branches, Tongbok and Bosung stream of Lake Chuam, Korea pennisula was examined. It showed extinct differences in most physicochemical factors between both branches, except water temperature and pH values. The concentrations of total phosphorus in Tongbok branch were twice as those of Bosung. M. aeruginosa cells were enumerated totally $1.2X10^4cells/ml$ and these individuals in branch of Tongbok were close to two times as much as Bosung. In light and electron microscopy, natural M. aeruginosa colonies formed irregular shape and non-directional array in amorphous matrix. They were consisted of many kinds of cells, youngs or olds in cell division, solitary, and various size of cells. Each cell ranged from 2.61 to $5.40{\mu}m$ in diameter, and averaged as $3.54{\pm}0.19{\mu}m$. In cytoplasm, they contained a number of inclusions in various size, shape and appearances. Among them, polyhedral bodies or carboxysomes, a structured granules, photosynthetic lamellae or thylakoids, and gas vacuoles were prominent and easy to recognize. Although it was failed to find the definable morphological variations in the ultrastructure of M. aeruginosa in terms of algal habitual environments, some useful characters were founded, outer layer of cell wall, polyhedral bodies and gas vacuoles, in blue-green algal classification and taxonomy.

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Effect of Zedoariae rhizoma on Bronchial Inflammation and Allergic Asthma in Mice

  • Ahn, Jong-Chan;Ban, Chang-Gyu;Park, Won-Hwan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.6
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    • pp.1636-1648
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    • 2006
  • There are detailed descriptions of the clinical experiences and prescriptions of asthma in traditional Korean medicine. Zedoariae rhizoma is one of the Korean herbal medicines used to treat bronchial asthma and allergic rhinitis for centuries. However, the therapeutic mechanisms of this medication are still far from clear, In this study, a house-dust-mite (Dermatophagoides pteronyssinus [Der p])-sensitized murine model of asthma was used to evaluate the immunomodulatory effect of Zedoariae rhizoma on the allergen-induced airway inflammation in asthma. Three different protocols were designed to evaluate the treatment and/or long-term prophylacitic effect of Zedoariae rhizoma in Der p-sensitized mice. Cellular infiltration and T-cell subsets in the bronchoalveolar lavage fluid (BALF)of allergen-challenged mice were analyzed. Intrapulmonary lymphocytes were also isolated to evaluate their response to allergen stimulation. When Zedoariae rhizoma was administered to the sensitized mice before AC (groups A and C), it suppressed airway inflammation by decreasing the number of total cells and eosinophil infiltration in the BALF, and downregulated the allergen- or mitogen-induced intrapulmonary lymphocyte response of sensitized mice as compared to those of controls. This immunomodulatory effect of Zedoariae rhizoma may be exerted through the regulation of T-cell subsets by elevation or activation of the CD8+ and double-negative T-cell population in the lung. However, the administration of Zedoariae rhizoma to sensitized mice 24 h after AC (group B) did not have the same inhibitory effect on the airway inflammation as Zedoariae rhizoma given before AC. Thus, the administration of Zedoariae rhizoma before AC has the immunomodulatory effect of reducing bronchial inflammation in the allergen-sensitized mice. On the other hand, to determine the potentiality of prophylactic and/or therapeutic approaches using a traditional herbal medicine, Zedoariae rhizoma, for the control of allergic disease, we examined the effects of oral administration of Zedoariae rhizoma on a murine model of asthma allergic responses. When oral administration of Zedoariae rhizoma was begun at the induction phase immediately after OVA sensitization, eosinophilia and Th2-type cytokine production in the airway were reduced in OVA-sensitized mice following OVA inhalation. These results suggest that the oral administration of Zedoariae rhizoma dichotomously modulates allergic inflammation in murine model for asthma, thus offering a different approach for the treatment of allergic disorders.

In Vivo Development of Mouse IVF/IVC Embryo Treated with Epidermal Growth Factor (EGF) (EGF 처리를 받은 체외생산된 생쥐배의 체내 발달)

  • Kim, E.Y.;Kim, M.K.;Yi, B.K.;Lee, H.S.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.2
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    • pp.261-265
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    • 1997
  • The objective of this study was to examine the effect of EGF to the in vivo development of mouse IVF/IVC embryo. The 2-cell embryos were cultured in medium (5-6 embryos/25 ${\mu}l$/drop) w/wo EGF (10 ng/ml) and day 4 blastocysts recovered from each treatment group were transferred into the uteri of recipients of pseudopregnant day 3. The results obtained in this experiment were summarized as follows: 1. When the effect of EGF to the in vitro development and cell number of blastocysts produced from the culture of 2-cell embryos in w/wo EGF was determined, those results of EGF treatment group showed not significant difference compared with control. 2. However, when the effect of EGF to the in vivo development of blastocysts recovered from each treatment group was examined, production of the normal fetus against transferred embryos in EGF treatment group (51.2%) was very higher than that in control group (31.1%), although total implantation was not significantly different between treatment group (control: 64.4%, EGF: 69.8%). Therefore, this result suggested that EGF can affect to the in vivo development of IVF/IVC embryos through the improvement of embryo quality, although EGF treated embryos showed not significant development rate compared with control.

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Effect Naetakchungumsankamibang on Skin tumor induced by 3-MCA and Immunological Response (內托千金散加味方이 3-MCA로 誘發된 皮膚癌 및 免疫調節作用에 미치는 影響)

  • Kim, Hee-taek;Roh, Seok-seon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.12 no.2
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    • pp.20-52
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    • 1999
  • In order to investigate the effect of Naetakchungumsankamibang(NTCGS) water extract on the skin tumor induced by 3-MCA and immunological responses in mice, the cytotoxicity against SK-MEL-2 cells and total number of tumors induced by 3-MCA were measured. The numbers of WBC, platelets and RBC, plaque forming cells, hemagglutinin titer, hemolysis titer, carbon clearance, proliferation of splenocyte by thymidine uptake assay, splenic leukocyte by FACS analysis and $TNF-{\alpha}$ were also measured for the evaluation of the immunological responses. The results were obtained as follows: 1. In cytotoxicity against SK-MEL-2 cells, concentration inhibiting cell growth up to below $20\%$ of control was recognized at 1mg/ml of NTCGS. 2. In Inhibitory effect on the skin tumor induced by 3-MCA, the results showed a strong inhibitory effect of NTCGS. 3. In hematological changes in the tumor bearing mice, the numbers of WBC decreased significantly in NTCGS treated group as compared with control. 4. In hematological changes in the tumor bearing mice, the numbers of platelets increased significantly in NTCGS treated group as compared with control. 5. In hematological changes in the tumor bearing mice, the numbers of RBC increased with no significance in NTCGS treated group as compared with control. 6. Effects of the plaque forming cells in the tumor bearing mice, NTCGS treated group exhibited a significant effect compared with control. 7. In terms of the effects on hemagglutinin titer, NTCGS treated group showed higher level than control, without significance. 8. In terms of the effects on hemolysis titer, NTCGS treated group showed higher level than control, without significance. 9. In terms of the effects on phagocytic index K in Balb/C mice, NTCGS treated group showed significant difference from control. 10. In terms of the effects on proliferation of splenocyte by thymidine uptake assay, NTCGS showed significant effect at the concentration of 0.5mg/ml. 11. In terms of the effects on splenic leukocyte of Balb/C mice by FACS analysis, NTCGS treated group showed significantly higher level of helper T cell, B cell and macrophage than in control. 12. In terms of the effects on the secretion of $TNF-{\alpha}$, the treated group showed significant effect at the concentration of 1mg/ml of NTCGS. Based on the results summarized above, NTCGS is considered to have antitumor activity and immunological responses against skin tumor, and to be usable fur the treatment.

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Mixture Bombyx mori L. and Liriopis seu Ophiopogonis Tuber effects on T cells in Thymus, Lymph Nodes (누에, 맥문동복합물의 흉선 림프절내 T세포 활성 유도 효능 연구)

  • Kim, Il Gyu;Park, Hae-Jin;Kim, Kyeong Jo;Kim, Soo Hyun;Kim, Min Ju;Lee, Jin A;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.33 no.5
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    • pp.47-52
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    • 2018
  • Objectives : The purpose of this study is to investigate effects on the immune system of Bombyx mori L. and Liriopis seu Ophiopogonis Tuber mixture (BL) in Thymus, Lymph Nodes. Methods : Eight-week-old male Balb/c mice were divided into five groups : Group one included the normal mice (Nor). Positive control group two administrated with red ginseng (RG) 100 mg/kg. Group three administrated with Bombyx mori L. (BX) 300 mg/kg. Group four administrated with Liriopis seu Ophiopogonis Tuber (LP) 300 mg/kg. Group five administrated with the mixture of Bombyx mori L. and Liriopis seu Ophiopogonis Tuber (BL) 300 mg/kg. After 2 weeks administration, mice were sacrified and antigen receptor in Thymus, Lymph Nodes was analyzed by using Fluorescence Activated Cellorter Sorting (FACS). we counted the total of Thymus and Lymph Nodes cells. GOT (glutamlc oxaloacetic transaminase), GPT (glutamlc pyruvic transamlnase) in serum were analyzed after experiment. Results : In Effects of Nor, RG, BX, LP, BL on the ratio of CD4+CD8+, CD4+CD69+ and CD4+CD25+ T cell in Thymus and Lymphnode, BL is higher than other groups except Nor in CD4+, CD4+CD69+, CD4+CD25+ T cell. The number of Thymus and Lymph Nodes increased in BL. In the level of GOT and GPT, BL decreased comparing to others group except Nor. Conclusions : BL may have effect on T cells in Thymus, Lymph Nodes. In addition, Bombyx mori could be immune functional material with others herb materials.

The Effect of the Timing and Dose of Human Chorionic Gonadotropin on Oocyte Recovery, in Vitro Fertilization, and Preimplantation Development in Superovulation of Mouse (생쥐에서 과배란 유도시 인간융묘 성선자극 홀몬 투여 방법이 체외수정 및 배자의 체외성장에 미치는 영향에 관한 연구)

  • Yang, S.H.;Kim, H.M.;Oh, S.H.;Son, Y.S.;Yoo, H.K.;Woo, B.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.2
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    • pp.165-176
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    • 1994
  • This study was carried out to investigate the effect of the timing and dose of human chorionic gonadotropin(hCG) on oocyte recovery, in vitro fertilization, and preimplantation development in superovulation of mouse. F1 hybrid($C57BL{\times}CBA$) mice were obtained and superovulation was induced in female mice by sequential intraperitoneal injection of PMSG and hCG. In the first series of experiments, mice received 5 IU of PMSG given intraperitoneally, and 48 hours later were injected 1 IU, 5 IU, or 10 IU of hCG respectively. In the second series of experiments, mice received 5 IU of PMSG given intraperitoneally and were injected 5IU of hCG 36, 48, or 60 hours later respectively. 1. When the mice received 5 IU of PMSG given intraperitoneally and 48 hours later were injected 1 ItT, 5 IU, or 10 IU of hCG respectively, there were no differences in the total number of the oocytes obtained from the three experimental groups. When the cultures were examined 48 hrs after the termination of insemination the proportion of unfragmented oocytes which had developed over two-cell stage was observer to be lowest in 10 IU hCG group. When the cultrues were examined 120 hour after termination of insemination the proportion of embryos which had developed to the blastocyst stage was observed to be significantly higher in 10IU hCG group than 5IU hCG group(p<0.05), but there was no difference between 10 IU hCG group and 1IU hCG group. 2. When the mice received 5 IU of PMSG and were injected 5 IU of hCG 36, 48, or 60 hours later respectively, there were no differences in the total number of oocytes obtained from the three experimental groups. When cultures were examined 48 hour after the termination of insemination the proportion of unfragmented oocytes which had developed over two-cell stage was observed to be significantly lower in 36 hour interval group than 48 hour interval and 60 hour interval group(p<0.05). When the cultures were examined 120 hour after termination of insemination the proportion of embryos which had developed to the blastocyst stage was found to be higher in 60 hour interval group than 36 interval or 48 hour interval group (P<0.05), and the proportion of hatching blastocyst was found to be higher in 60 hour interval group as well. In this study, it was concluded that the administration of adequate dose of hCG, and long (60 hour) PMSG-hCG interval were necessary in superovulation of mice($C57BL{\times}CBA$) in order to get a large number of oocytes which had an early oocytes which had an early embryonic developmental capability when fertilized in vitro, and especially it had better have been avoided to administer a large dose of hCG.

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A Tetraploid Induction in Hypericum patulum Thunberg by Colchicine Soaking Treatment (콜히친 침지처리에 의한 '망종화'의 4배체 식물유도)

  • Kwon, Soo Jeong;Cho, Kab Yeon;Kim, Hag Hyun
    • Korean Journal of Plant Resources
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    • v.26 no.2
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    • pp.284-288
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    • 2013
  • This study aimed to get the basic data on the breeding of good varieties in Hypericum patulum Thunberg. The optimum materials, concentration and soaking time were examined to identify the effective approach to induce tetraploid plant by colchicine treatment to cultivate the varieties. For the seed germination rate of seed by colchicine treatment, the higher colchicine concentration was and the longer soaking time was, the more the germination rate decreased. While individuals were germinated in 16 test groups except control group (no treatment group), all the plants were diploid and no tetraploid was induced. For the plant regeneration rate by colchicine treatment on the explant of Hypericum patulum Thunberg that was under in vitro culture, the higher the colchicine concentration increased, the ress the regeneration rate. While total 147 individuals were regenerated in all treatment, when the explant was soaking treatment in more than 0.05% for over 6 hours, tetraploid could be obtained. In the soaking treatment of 0.05% for over 6 hours, tetraploid could be obtained. In particular, for the soaking treatment in 0.05% for 12 hours, 8 tetraploids were induced, which was about 47.1% of the number of plant regenerated. In accordance with the observation on doubling of DNA contents in leaf in order to identify polyploidy, the peak DNA content of G1 phase was 94.5 for diploid and 192.5 for tetraploid. It confirmed doubling of DNA content. Furthermore, the number of chloroplasts per guard cell depending on polyploid was around 10 in diploid and 17 to 19 in tetraploid, which were around 1.7 to 1.9 times as much as diploid.

Maximization of The Number of Follicular Oocytes Recovered from The Bovine Ovaries (소 난소로부터 회수난포란수의 극대화 방법)

  • 유형진;최승철;이상호
    • Korean Journal of Animal Reproduction
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    • v.17 no.2
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    • pp.149-157
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    • 1993
  • A new technique was established to maximize the numbers of follicular oocytes recovered from the ovaries obtained at the slaughter house. And their further developmental capacity was demonstrated. There recovery techniques were used; aspiration (ASP, control), slicing (SLC) and slicing combining aspiration (ASP+SLC). Recovered oocytes were cultured in TCM 199+15% FCS+gonadotrophins in an atmosphere of 5% CO$_2$ in air at 39$^{\circ}C$ for 24 h. The nuclear maturation was detemined with chromo-some configuration by rapid staining. And cytoplasmic maturation was examined by the formation of female pronuclei with parthenogenetic activation of the matured oocyte after 18 h of co-culture with granulosa cell monolayer. Total 1,641 bovine follicular oocytes recovered from 245 ovaries. The number of oocytcs per ovary was 1.87 in ASP, 11.05 in SLC and 7.88 in ASP+SLC, respectively. SLC would yield 5.9 folds increase, compared with ASP. The rate of maturation were 92.9% in ASP, 79.1% in SLC and 71.7% in ASP+SLC, respectively. Although the maturation rate in ASP was the highest, metaphase II oocytes per ovary in SLC was 5 times higher than that of ASP. The rates of pronuclei formation upon ethanol activation were 75% in ASP, 67% in SLC and 62.5% in ASP+SLC, respectively. The results demonstrate that it should be possible to maximize the number of the follicular oocyte from the ovary for mass production of bovine embryos. Thus the established technique may provide efficient supply of bovine embryos for biochemical and molecular study of early bovine embryos.

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Hardware optimized high quality image signal processor for single-chip CMOS Image Sensor (Single-chip CMOS Image Sensor를 위한 하드웨어 최적화된 고화질 Image Signal Processor 설계)

  • Lee, Won-Jae;Jung, Yun-Ho;Lee, Seong-Joo;Kim, Jae-Seok
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.44 no.5
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    • pp.103-111
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    • 2007
  • In this paper, we propose a VLSI architecture of hardware optimized high quality image signal processor for a Single-chip CMOS Image Sensor(CIS). The Single-chip CIS is usually used for mobile applications, so it has to be implemented as small as possible while maintaining the image quality. Several image processing algorithms are used in ISP to improve captured image quality. Among the several image processing blocks, demosaicing and image filter are the core blocks in ISP. These blocks need line memories, but the number of line memories is limited in a low cost Single-chip CIS. In our design, high quality edge-adaptive and cross channel correlation considered demosaicing algorithm is adopted. To minimize the number of required line memories for image filter, we share the line memories using the characteristics of demosaicing algorithm which consider the cross correlation. Based on the proposed method, we can achieve both high quality and low hardware complexity with a small number of line memories. The proposed method was implemented and verified successfully using verilog HDL and FPGA. It was synthesized to gate-level circuits using 0.25um CMOS standard cell library. The total logic gate count is 37K, and seven and half line memories are used.