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.
Lee Suk;Lee Sang Hoon;Shin Dongho;Yang Dae Sik;Choi Myung Sun;Kim Chul Yong
Radiation Oncology Journal
/
v.22
no.4
/
pp.316-324
/
2004
Purpose : In radiotherapy of tumors in liver, enough planning target volume (PTV) margins are necessary to compensate breathing-related movement of tumor volumes. To overcome the problems, this study aims to obtain patients' body movements by using a moving phantom and an ultrasonic sensor, and to develop respiration sating techniques that can adjust patients' beds by using reversed values of the data obtained. Materials and Methods : The phantom made to measure patients' body movements is composed of a microprocessor (BS II, 20 MHz, 8K Byte), a sensor (Ultra-Sonic, range $3\~3$ m), host computer (RS232C) and stepping motor (torque 2.3 Kg) etc., and the program to control and operate it was developed. The program allows the phantom to move within the maximum range of 2 cm, its movements and corrections to take place In order, and x, y and z to move successively. After the moving phantom was adjusted by entering random movement data (three dimensional data form with distance of 2 cm), and the phantom movements were acquired using the ultra sonic sensor, the two data were compared and analyzed. And then, after the movements by respiration were acquired by using guinea pigs, the real-time respiration gating techniques were drawn by operating the phantom with the reversed values of the data. Results : The result of analyzing the acquisition-correction delay time the three types of data values and about each value separately shows that the data values coincided with one another within $1\%$ and that the acquisition-correction delay time was obtained real-time $(2.34{\times}10^{-4}sec)$. Conclusion : This study successfully confirms the clinic application possibility of respiration gating techniques by using a moving phantom and an ultrasonic sensor. With ongoing development of additional analysis system, which can be used in real-time set-up reproducibility analysis, it may be beneficially used in radiotherapy of moving tumors.
Natural orifice transluminal endoscopic surgery is a newly emerging technique recently, with its many potential advantages in clinical practice. Cholecystectomy by Hybrid NOTES in this work, performed with single working channel endoscope in conjunction with a laparoscopic grasping forceps in dogs, is a "bridge" between laparoscopic procedure and pure NOTES. Three different approaches for cholecystectomy were carried out; transgastric, transcolonic and transvaginal. In all three approaches, abdominal opening was made by a 5 mm trocar, followed by making pneumoperitoneum of 4 mmHg with $CO_2$ insufflator. Transgastric cholecystectomy, single working channel endoscope was advanced to the peritoneal cavity through gastric incision in antral region made by endoscopic needle knife. Endoscope was retroflexed to visualize the gall bladder. Transcolonic access, incision for endoscopic entry was done at right ventral wall of descending colon, 15 cm inside from the anus. Incision in transvaginal access was made at right-ventral region, just caudal to the caudal tubercle. With the simple traction by the laparoscopic grasping forceps, good visualization of surgical field was obtained in all three groups. Cystic duct and artery were ligated with endoclips; for complete gall bladder dissection from liver, L-knife was used. Closure of incision sites were done in transgastric and transcolonic cholecystectomy by endoclips, not in transvaginal approach.
Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.
A series of experiments on the effects of ${\gamma}-irradiation$ was performed to reveal the pathogenicity of ${\gamma}-irradiated$ oocysts of E tenella from Cobalt-60 and its progeny. The SPF chickens were inoculated with differnt doses of radiation and inoculum. The level of 100 Gy ${\gamma}-irradiation$ from $^{60}Co$ and the level of inoculum with $1{\times}10^4$ oocysts were recognized more pathogenic than those of the other groups by comparison of body weight gains, blood in feces and lesion scores. The signs of blood in feces, lesion score and the number of excreted oocysts in the feces were revealed as the lowest in the group of the ${\gamma}-irradiated$ oocysts, the average in the group of the 1st and the 3rd progeny, and the highest in the group of non-irradiated oocysts of E $tenell\grave{a}$. The body weight gain of the group immunized with ${\gamma}-irradiated$ oocysts of E tenella was higher than those of the non-irradiated, the 1st and 3rd progeny groups. The body weight gain of the groups immunized with the 1st and the 3rd progeny of E tenella were higher than that of the non-irradiated group. The feed conversion ration of the group immunized with ${\gamma}-irradiated$ oocysts of E tenella was lower than those of the non-irradiated, the 1st and the 3rd progeny groups. The feed conversion ratios of the group immunized with the 1st and 3rd progeny of ${\gamma}-irradiated$ oocysts were lower than that of the group infected with non-irradiated E tenella. The anticoccidial index(ACI 190.6) in the chickens immunized with the ${\gamma}-irradiated$ oocysts of E tenella and those(ACI 142.8 and 107.4) of the 1st and the 3rd progeny groups were higher than that (ACT 87.4) of the group infected with non-irradiated E tenella. It was thought that the pathogenicity of ${\gamma}-irradiated$ E tenella would be recovered according to increase the number of generation passaged in chicken.
This study was conducted to evaluate the effect of a growth retardant, Paclobutrazol on the growth, yield and its components. and physiological traits in peanut cv. Saeddle (early variety) and Nampung (medium variety). The results are summarized as follows : Main stem and branch length of both varieties were remarkably retarded at early growth stage application, but the retardation effect was reduced at late growth stage application. Number of branches was increased remarkably by treating 60ppm at 20days after seedling (DAS) in early variety. Lodging ill medium variety was not observed through growth period when 120ppm of Paclobutrazol was applied while lodging was not appeared between 40 DAS and 80 DAS in early variety. The numbers of riped pods in early variety was increased when application of 120ppm at 40 DAS was made while it was increased as application of 120ppm at 90 DAS in the medium variety. Cercospora leaf spot was reduced by spraying at 40 or 50 DAS regardless of concentration and varieties. Chlorophyll content and photosynthetic activity increased when paclobutrazol was applied during 40-60 DAS in early variety while these were observed at 90 DAS in medium variety. Seed weight tended to increase as paclobutrazol application was delayed. The ratio of pods to seeds was higher at 60ppm-40 DAS in early variety while it was higher at 120ppm-90 DAS in medium variety.
This survey of weed population by different soil moisture with paddy-upland rotation was conducted to investigate information on weed flora and its ecology between two years. Weed species were assessed in April : 2009, 2010. Soil samples were taken from each study plot to assess the impact soil moisture on the occurrence and abundance of dominance weed species. Soil water of poorly drained field ranged from 10.2~18.2% more than 11.8~14.3% of somewhat poorly drained field. Weeds of fields composed of 19 species belonging to 12 families. Among 12 families, 6 weed species belonged to Compositae (31.6%) were the biggest family, Cruciferae were 2 species (10.5%), and Rubiaceae were 2 species (10.5%), respectively. Youngia japonica had the highest summed dominance ratio(SDR) (97.0%) and followed by Conyza canadensis (92.1%), Galium spurium (35.3%) and Hemistepta lyrata (28.4%) of somewhat poorly drainage in 2009. Artemisia princeps the highest SDR (100%) and followed by Stellaria alsine(55.2%), Y. japonica (38.3%) and Nasturtium officinale (28.5%) of poorly drainage in 2009. And, Stellaria alsine had the highest SDR (86.8%) and followed by Alopecurus aequalis (77.7%), Astragalus sinicus (68.7%) and Y. japonica (46.3%) of somewhat poorly drainage in 2010. S. alsine the highest SDR (93.7%) and followed by A. aequalis (78.6%), Nasturtium officinale (31.3%) and Y. japonica (30.4%) of poorly drainage in 2010. Simpson's index was calculated to 0.12~0.23, which showed that weed occurrence with different soil moisture in paddy-upland rotation and between years was various. Similarity coefficient between years was 43.0% (2009) and 74.2% (2010), which indicate a low diversity because of the moisture in the agro-ecosystem.
Ettringite $(Ca_6[Al(OH)_6]_2(SO_4)_3{\cdot}26H_2O)$ is a sulfate mineral that shows a complicate property in concrete. It is often called as "a cancer of concrete" because secondary ettringite formation in hardened concrete often cause expansion and cracking of concrete due to its expansive crystal structure. In the present study, we tested the possibility for crystal growth inhibition of secondary ettringite by crystallization inhibitors that are commercially used for scaling inhibitors in Korea. For the test, we developed a method of ettringite solution synthesis. Three types of crystallization inhibitors were selected and examined the effects On ettringite growth inhibition. The experimental results of ettringite solution synthesis indicated that ettringite was successfully synthesized under condition that the mass balance between calcium hydroxide saturated solution and aluminum sulfate solution was attained. Monosulfate and semisulfate were synthesized when the ratio of $Ca^{2+}$ ions to ${SO_4}^{2+}$ ions was increased. The induction time of ettringite crystallization was less than 2 min. and crystallization was almost completed within an hour. The experimental results of ettringite crystallization inhibition showed that organic PBCT (2-Phosphonobutane-1,2,4-Tricarboxylic Acid) and inorganic SHMP (Sodium Hexametaphosphate) were relatively less effective on ettringite crystallization inhibition under experimental conditions. However, organic HEDP (1-Hydoxyethylidene-1,1-Diphosphonic Acid) effectively prevented ettringite growth with producing amorphous gel phase materials up to inhibitor concentration 0.1 vol.% of aluminum sulfate solution.
Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.
Cheongpung Buwongun Kim Wu-myeong's Funeral Bier, an important folklore cultural property No.120, possessed by Chuncheon National Museum was donated in 2002 (by Kim Seonggu). It consists of a bier, yoyeo(腰輿), myeongjeongdae(銘旌臺), and manjangdae(輓章臺). It has a high value as the oldest royal bier. The bier which had a resting time in the storage for special exhibition of "The great cultural treasure of Gangwon province" was inspected in September 2012 and colored pigment layer of the wooden part had the risk of peeling off and surface damage of the textile was serious. Therefore, conservation treatment was conducted. In addition, knots and susiks(垂飾) were severely damaged and their exhibition was impossible. Therefore, a reproduction to replace them through a close investigation was made. All parts of the funeral bier were in separation except for the basic furniture. Conservation was made by dividing the parts into wooden parts and textile parts. Yoyeo was reinforced after disassembling bujae from it and then was reassembled. Paraloid B-72 2 wt% (in ethyle acetate), acrylic resin, was applied to the wooden part of the bier in order to reinforce the colored pigment layer with the addition of sodium alginate 2 wt%(in stilled water) and glue 4 wt%(in stilled water). The pollutants on the surface of the textile part were removed (vacuuming) and its creases were smoothed out (steaming). Fat-soluble pollutants were removed using an nonionic surfactant(Saponin, concentration at 0.25 to 0.5 g/𝑙, in de-ionized water). After the disassembly of the yoyeo from the broken wooden, it was bonded with glue (3 wt% for the first gluing, 35 wt% for gluing), and pine wood was used to restore missing parts. In the process of connecting Wongak(雲角), the original metal hinge and nails were reused to complete the assembly.
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