• Title/Summary/Keyword: biological study

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Improvement of Proliferation Capacity of Non-adapted CHO Cells Subcultured Using Serum Free Media in Long-term Culture (무혈청 배지에서 계대배양한 비적응 CHO(Chinese Hamster Ovary) 세포의 증식력 개선에 관한 연구)

  • Lee, Seung-Sun;Lee, Jin-Sung;Byun, Soon-Hyu;Park, Hong-Woo;Choe, Tae-Boo
    • KSBB Journal
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    • v.21 no.4
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    • pp.248-254
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    • 2006
  • Animal cell culture industry has a large market and an exponential growth rate among biological industry field. Chines hamster ovary(CHO) cells are the most widely used cell lines for recombinant protein production. They can avoid infection from polio, herpes, hepatitis B, HIV, measles, adenovirus and etc. Moreover it is easy to transfection recombinant genes and possible to suspension culture. Serum free media is one of the most important factor of protein production. Because serum has problems. Serum is not defined the contents until now, it has a number of proteins, lipids, carbohydrates and unknown molecules that cause of risk involve in infection and high cost of product purification. CHO cell line cultured using serum free media were the basis of a very successful method to produce(glyco-)protein in mammalian cells, which are then used as pharmaceutical products. Also, the low protein content of the developed medium facilitates downstream processing and product purification. But non-adapted CHO cells have a limit of proliferation cultured using serum free media and it takes very long time to adapt non-adapted cells to serum free media. There are a number of causes of a limit of proliferation using serum free media. Absence of growth factors and growth stimulating molecules is a major factor of the reasons. It makes growth signals and moves cell cycle. And increase of cellular stress is another reason. It induces increase of intraceullar ROS concentration. The purpose of this study is about improvement of proliferation capacity of non-adapted CHO cells cultured using serum free media without adaptation process.

Trend in Research and Application of Hard Carbon-based Thin Films (탄소계 경질 박막의 연구 및 산업 적용 동향)

  • Lee, Gyeong-Hwang;Park, Jong-Won;Yang, Ji-Hun;Jeong, Jae-In
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2009.05a
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    • pp.111-112
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    • 2009
  • Diamond-like carbon (DLC) is a convenient term to indicate the compositions of the various forms of amorphous carbon (a-C), tetrahedral amorphous carbon (ta-C), hydrogenated amorphous carbon and tetrahedral amorphous carbon (a-C:H and ta-C:H). The a-C film with disordered graphitic ordering, such as soot, chars, glassy carbon, and evaporated a-C, is shown in the lower left hand corner. If the fraction of sp3 bonding reaches a high degree, such an a-C is denoted as tetrahedral amorphous carbon (ta-C), in order to distinguish it from sp2 a-C [2]. Two hydrocarbon polymers, that is, polyethylene (CH2)n and polyacetylene (CH)n, define the limits of the triangle in the right hand corner beyond which interconnecting C-C networks do not form, and only strait-chain molecules are formed. The DLC films, i.e. a-C, ta-C, a-C:H and ta-C:H, have some extreme properties similar to diamond, such as hardness, elastic modulus and chemical inertness. These films are great advantages for many applications. One of the most important applications of the carbon-based films is the coating for magnetic hard disk recording. The second successful application is wear protective and antireflective films for IR windows. The third application is wear protection of bearings and sliding friction parts. The fourth is precision gages for the automotive industry. Recently, exciting ongoing study [1] tries to deposit a carbon-based protective film on engine parts (e.g. engine cylinders and pistons) taking into account not only low friction and wear, but also self lubricating properties. Reduction of the oil consumption is expected. Currently, for an additional application field, the carbon-based films are extensively studied as excellent candidates for biocompatible films on biomedical implants. The carbon-based films consist of carbon, hydrogen and nitrogen, which are biologically harmless as well as the main elements of human body. Some in vitro and limited in vivo studies on the biological effects of carbon-based films have been studied [$2{\sim}5$].The carbon-based films have great potentials in many fields. However, a few technological issues for carbon-based film are still needed to be studied to improve the applicability. Aisenberg and Chabot [3] firstly prepared an amorphous carbon film on substrates remained at room temperature using a beam of carbon ions produced using argon plasma. Spencer et al. [4] had subsequently developed this field. Many deposition techniques for DLC films have been developed to increase the fraction of sp3 bonding in the films. The a-C films have been prepared by a variety of deposition methods such as ion plating, DC or RF sputtering, RF or DC plasma enhanced chemical vapor deposition (PECVD), electron cyclotron resonance chemical vapor deposition (ECR-CVD), ion implantation, ablation, pulsed laser deposition and cathodic arc deposition, from a variety of carbon target or gaseous sources materials [5]. Sputtering is the most common deposition method for a-C film. Deposited films by these plasma methods, such as plasma enhanced chemical vapor deposition (PECVD) [6], are ranged into the interior of the triangle. Application fields of DLC films investigated from papers. Many papers purposed to apply for tribology due to the carbon-based films of low friction and wear resistance. Figure 1 shows the percentage of DLC research interest for application field. The biggest portion is tribology field. It is occupied 57%. Second, biomedical field hold 14%. Nowadays, biomedical field is took notice in many countries and significantly increased the research papers. DLC films actually applied to many industries in 2005 as shown figure 2. The most applied fields are mold and machinery industries. It took over 50%. The automobile industry is more and more increase application parts. In the near future, automobile industry is expected a big market for DLC coating. Figure 1 Research interests of carbon-based filmsFigure 2 Demand ratio of DLC coating for industry in 2005. In this presentation, I will introduce a trend of carbon-based coating research and applications.

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IL-1 AND TNF-α RELEASE IN HUMAN POLYMORPHONUCLEAR LEUKOCYTES AFTER EXPOSURE TO CALCIUM HYDROXIDE TREATED Porphyromonas endodontalis LIPOPOLYSACCHARIDE (수산화칼슘 처리된 Porphyromonas endodontalis Lipopolysaccharide가 다형핵백혈구의 IL-1과 TNF-α 생성에 미치는 영향에 관한 연구)

  • Park, Chan-Je;Park, Dong-Sung;Yoo, Hyeon-Mee;Oh, Tae-Seok;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.27 no.5
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    • pp.463-472
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    • 2002
  • Bacterial lipopolysaccharide (LPS) plays a major role in stimulating the synthesis and release of the principal osteoclast-activating cytokines, namely, interleukin 1 and tumor necrosis factor-$\alpha$ from immune cells. Although rnonocytes/macrophages are the main producers of these cytokines, recent evidence has indicated that polymorphonuclear leukocytes (PMN) have the ability to release IL-1 and TNF-$\alpha$. Calcium hydroxide has been shown to be an effective medicament in root canal infections, reducing the microbial titre within the canal. It has been proposed that the therapeutic effect of Ca(OH)$_2$ may also be the result of direct inactivation of LPS. The purpose of this study was to investigate whether treatment of Porphyromonas endodontalis LPS with calcium hydroxide alters its biological action as measured by human PMN secretion of IL-1 and TNF-$\alpha$, and it was compared with Escherichia coli LPS. P. endodontalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted using the hot-phenol water extraction method and purified. Purchased E. coli LPS was also purified. 100 $\mu\textrm{g}$/ml of each LPS in pyrogen free water were incubated with 25mg/ml Ca(OH)$_2$ at 37$^{\circ}C$ for 7 days. The supernatants were subjected to ultrafiltration, and the isolates were lyophilized and weighed. PMNs were obtained from peripheral blood by centrifugation layered over Lymphoprep. The cells were resuspended (4$\times$10$^6$ cells/ml) in RPMI 1640 followed by treatment with various concentrations of LPS (0, 0.1, 1, 10$\mu\textrm{g}$/ml) for 24 hours at 37$^{\circ}C$ in 5% $CO_2$ incubator. The supernatants of cells were collected and the levels of IL-1$\alpha$, IL=1$\beta$ and TNF-$\alpha$ were measured by enzyme-linked immunosorbent assay. The results were as follows ; 1. The levels of IL-1$\alpha$, IL-1$\beta$, TNF-$\alpha$ from PMN treated with each LPS were significantly higher than those released from unstimulated PMN of the control group (p<0.05). 2. The levels of all three cytokines released from PMN stimulated with each calcium hydroxide treated LPS were significantly lower than those released from PMN stimulated with each untreated LPS (p<0.05), while they were not significantly different from those released from unstimulated PMN of the control group (p>0.05) 3. The levels of secretion for all three cytokines were affected in a dose-dependent manner in PMN stimulated with each LPS (p<0.05), but not in PMN stimulated with each calcium hydroxide treated LPS (p>0.05). 4. The levels of all three cytokines released from PMN stimulated with p. endodontalis LPS were significantly lower than those released from PMN stimulated with E coli LPS (p<0.05).

Antioxidant Activity of Solvent Extracts from Sargassum thunbergii (지충이 용매 추출물의 항산화 활성)

  • Choi, Sun-Young;Kim, Soon-Young;Hur, Jong-Moon;Choi, Han-Gil;Sung, Nak-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.2
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    • pp.139-144
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    • 2006
  • In this study, we investigated the antioxidant activity of solvent extracts from Sargassum thunbergii by measuring electron-donating ability (EDA), nitrite- scavenging ability, superoxide dismutase (SOD)-like activity, reducing power and antioxidant activity in $\beta-carotene-linoleate$ model system. Total phenolic compound contents of chloroform and methanol extracts from Sargassum thunbergii were $22.5{\pm}0.8$ mg/100 mL and $20.1{\pm}0.6$ mg/100 mL, respectively. The EDA were proportionally increased with concentration of the extracts except hexane extract. The EDAs were $70.4\%\;and\;63.2\%$ in methanol and water at concentration of $1000{\mu}g/mL$, respectively. The nitrite-scavenging ability of methanol extract (pH 2.5, $1000{\mu}g/mL$) was $89.4{\pm}0.9\%$ that was higher than those of other extracts. SOD-like ability of chloroform extract was stronger than other extracts. Reducing power of chloroform extracts at $1000{\mu}g/mL$ were 0.75 as O.D. value of 700 nm and antioxidant activities measured with $\beta-carotene-linoleate$ model system were in order of methanol $(78.4\%)>BHT\;(72.1\%)>chloroform\;(62.2\%)>water\;(51.4\%)>hexane\;(43.2\%)$. These results indicated that methanol extract from Sargassum thunbergii showed the highest activities on the EDA, nitrate- scavenging ability and antioxidant activities while the chloroform extract had the highest effect on the SOD-like activity and reducing power.

Effect of X-Irradiation on the Levels of some Sulfhydryl Groups, Protein and Cell Volume of Ehrlich Ascites Tumour Cells (X-선(線) 조사(照射)가 Ehrlich 암세포(癌細胞)의 용적(容積), 단백양(蛋白量) 및 수종(數種) Sulfhydryl 기(基)에 미치는 영향(影響)에 관(關)하여)

  • Yu, Choon-Shik;Choo, Young-Eun
    • The Korean Journal of Physiology
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    • v.3 no.2
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    • pp.9-16
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    • 1969
  • It is well known that a number of -SH and -SS containing substances afford a certain measure of protection against radiation effects in many biological systems, and it is conceivable that inherent -SH levels in Ehrlich ascites tumour (ELD)cells may be of decisive improtance with respect to the development of cellular radiation injury. So far, little effort has been directed to elucidate the changes in levels of different -SH and -SS groups in ELD cells when the tumour-bearing whole animal was subjected to the sublethal dose of X-irradiation. The present study was designed to bring some lights in the possible changes of and relationship between various sulfhydryl levels, such as P-SH, NP-SH and NP-SS, as well as the content of protein and cell volume of ELD cells, after subjecting the ELD mice to 1,200 r of X-irradiation. The animals used in this experiment were all mixed bred mice of $20{\sim}25\;gm$ in body weight (approximately 2 months old) irrespective of sex. 12 mice in one experiment were inoculated intraperitoneally with 0.2 ml of ascites tumour cells $(2{\times}10^6\;cells)$, and on the 7th day of the tumour growth, they were X-irradiated with 1,200 r, using the conventional X-ray machine under the following conditions: 200 Kv at 15 mA, 0.5 mm Cu filter, target-skin distance: 50 cm. Radiation dose was measured with the the Philip integrating dosimeter. At 24, 36, 48 and 60 hours after the X-irradiation, the mice were killed by cervical dislocation, and the tumours were taken out. Freshly withdrawn ascites tumours were placed in ice, and immediately the cell concentration was measured with the Coulter Cell Counter (Model B), and the hematocrit of the tumour cells were also determined. Cell volume was thus calculated by the cell concentration and hematocrit value. P-SH content of ELD cells was measured potentiometrically according to the method of Calcutt & Doxey, and NP-SH and NP-SS contents were measured spectrophotometrically by the method described by Ellman. Protein content of ELD cells was determined with the Folin phenol reagent by Lowry et al. Altogether, 48 experimental mice were used, and 12 mice with the only exception of X-irradiation were used as the control. Results obtained indicate that the contents of all the cellular sulfhydryl groups as well as cell volume and protein content of the ELD cells increase significantly as time progresses after the sub-lethal X-ray dose of 1,200 r was given and that all the increase is in a lineal fashion. The regression lines of the relative values, (i. e., taking each control value as 1) of all the values obtained, and the regression lines of cell volume, protein and NP-SH are identical, whereas those of NP-SS and P-SH appear to be widely seperated. However, the difference of those two lines (NP-SS & P-SH) were found to be not significant statistically (p>0.05). Therefore, it can be concluded from the above results that all the values examined increase in a lineal fashion with no statistically significant difference among them. Also, with the radiation dose of 1,200 r, the ELD cell becomes enlarged and swollen progressively up to 60 hours post-irradiation and it becomes more than two times of the original normal size at 60 hours after the irradiation, and up to this stage, it seems apparent that the cell division has been slow due to the X-irradiation applied in this experiment. It is well understandable that the contents of NP-SH, NP-SS, P-SH and protein of the ELD cells increase in parallel with the increase of the cell volume by the X-ray does used, but it also seems interesting to note that all the cellular substances tested show no appreciable difference in the pattern of increase.

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Effects of Sea Tangle (Laminaria japonica) Extract and Fucoidan Drinks on Oxygen Radicals and Their Scavenger Enzymes in Stressed Mouse (스트레스 부하 마우스의 활성산소 및 제거효소에 미치는 다시마(Laminaria japonica)와 후코이단 음료의 영향)

  • CHOI Jin-Ho;KIM Dae-Ik;PARK Soo-Hyun;KIM Dong-Woo;KIM Chang Mok;KOO Jae Geun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.6
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    • pp.764-769
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    • 1999
  • This study was designed to investigate the effects of sea tangle (Laminaria japonica) extract (Dasi-Ex group: dry base $4.0\%$) and fucoidan-added (Fuco-I, II, III group: fucoidan of $1,0\%,\;2.0\%,\;3.0\%$ added to Dasi-Ex) drinks on the formation of oxygen radicals and scavenger enzyme activities of stressed mice. ICR male mice (20 $\pm$2 g) were fed experimental diets and these drinks instead of water for 18 days including 4 days of sociopsychological stress. Dasi-Ex and Fuco-I, II and III groups resulted in a marked decreases $20\~40\%$ in basal oxygen radical (BOR) formation, and $15\~25\%$ in induced oxygen radical (IOR) formation compared with control group. Hydroxyl radical formations were significantly inhibited about $10\%$ in Dasi-Ex group, while remarkably inhibited $30\~40\%$ in Fuco-I, II and III groups. lipid peroxide (ISO) levels in Dasi-Ex group were not significantly different from those of control group, tut Fuco-I, II and III groups resulted in a significant decreases about $10\%$ in LPO levels compared with control group, Dasi-Ex, Fuco-I, II and III groups resulted in a marked decreases ($31\%,\;36\%,\;39\%$ and $42\%$, respectively) in oxidized protein levels through production of carbonyl group. Significant differences in nitric oxide (NO) levels in Dasi-Ex group were not obtained, but NO levels were slightly inhibited about $7\%$ in Fuco-I and II groups and $20\%$ in Fuco-III group compared with control group. Significant differences in superoxide dismutase (SOD) and catalase (CAT) activities in Dasi-Ex and Fuco-I groups were not obtained, but Fuco-II and III groups resulted in a significant increases $25\~40\%$ in SOD activities, and about $10\%$ in CAT activities compared with control group. These results suggest that the sociopsychological stress and aging process could be effectively inhibited by biological activity of sea tangle and fucoidan components.

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Effect of Brown Algae (Undasia pinnatisda)-Noodle on Inhibitory Action of Obesity in SD Rats (미역 (Undasia pinnatisda) 국수가 SD계 흰쥐의 비만 억제작용에 미치는 영향)

  • CHOI Jin-Ho;KIM Dong-Woo;KIM Jung-Hwa;KIM Dae-Ik;LEE Jong-Soo;BAEK Yeong-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.1
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    • pp.46-49
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    • 1999
  • Our previous studies have demonstrated the biological effects of alginic acid as a brown algae (Undasia pinnatisda) component on inhibitory action of obesity using animal model. This study was designed to investigate the effect of brown algae (BA)-noodles ($10\%,\;20\%$ and $40\%$) on inhibitory action of obesity in Sprague-Dawley (SD) male rats for 4 weeks. Body weight gain in SD rats for 4 weeks resulted in a marked inhibition ($7\%,\;17\%\;38\%$, respectively) by administration of $10\%,\;20\%$ and $40\%$ BA-noodles compared with control group. Total energy intake and feed efficiency in SD rats for 4 weeks resulted in a marked inhibition ($6\%,\;16\%\;37\%$ and $10\%\;20\%\;40\%$, respectively) by administration of $10\%\;20\%$ and $40\%$ BA-noodles compared with control group, but gross efficiency was not observed in these BA-noodle groups. Rohrer index and TM index as an obesity markers in SD rats for 4 weeks significantly decreased ($10\%\;15\%$ and $22\%$, respectively) by administration of $10\%,\;20\%$ and $40\%$ BA-noodles compared with control group. These results suggest that inhibitory effect of obesity in SD rats for 4 weeks may be due to decreases of body fat content resulted in a decrease of feed efficiency and energy intake by these BA-noodles.

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Clinical effects of combination anorganic bovine-derived hydroxyapatite matrix(ABM)/cell binding peptide (P-15) in periodontal intrabony defects (치주 골내낭에서 anorganic bovine-derived hydroxyapatite matrix(ABM)/cell binding peptide(P-15)의 임상적효과)

  • Won, Mi-Sook;Paik, Jeong-Won;Kim, Chang-Sung;Cho, Kyoo-Sung;Chai, Jung-Kiu;Kim, Chong-Kwan;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.32 no.3
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    • pp.565-576
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    • 2002
  • The ultimate goal of periodontal therapy is the regeneration of periodontal tissue which has been lost due to destructive periodontal disease. To achieve periodontal regeneration, various kinds of methods have been investigated and developed, including guided tissue regeneration and bone graft. Bone graft can be catagorized into autografts, allografts, xenografts, bone substitutes. And materials of all types have different biological activity and the capacity for periodontal regeneration, but ideal graft material has not been developed that fits all the requirement of ideal bone graft material. Intensive research is underway to identity, purify, synthesize a variety biologic modulators that may enhance wound healing and regeneration of lost tissues in periodontal therapy. The present study evaluates the effects of ABM/P-15 on the periodontal regeneration in intrabony defects of human. We used thirty four 2-wall or 3-wall osseous defects in premolars and molars of chronic peridontitis patient that have more than 5mm pockets and more than 3mm in intrabony defect. 12 negative control group underwent flap procedure only, 11 positive control group received DFDBA graft with flap procedure, and 11 experimental group received ABM/P-15 graft with flap procedure. The changes of probing pocket depth, loss of attachment and bone probing depth following 6months after treatment revealed the following results: 1. The changes of probing pocket depth showed a statistically significant decrease between after scaling and 6months after treatment in negative control(2.0${\pm}$0.9mm), positive control(3.0${\pm}$0.9mm), and experimental group (3.4${\pm}$1.5mm) (P<0.01). Significantly more reduction was seen in experimental group compared to negative control group (P<0.05). 2. The changes of loss of attachment showed a statistically significant decrease between after scaling and 6months after treatment in positive control(2.0${\pm}$0.6mm), and experimental group (2.2${\pm}$l.0mm) except negative control group(0.1${\pm}$0.7mm) (P<0.01). Significantly more reduction was seen in both experimental and positive control group compared to negative control group(P<0.05). 3. The changes of bone probing depth showed a statistically significant decrease between after scaling and 6months after treatment in positive control(2.7${\pm}$l.0mm), and experimental group (3.4${\pm}$1.3mm) except negative control(0.l${\pm}$0.9mm) (9<0.01). Significantly more reduction was seen in both experimental and positive control group compared to negative control group (P<0.05). The results suggest that the use of ABM/P-15 in the treatment of periodontal intrabony defects can reduce loss of attachment and bone probing depth more than flap operation only. It suggests that ABM/P-15 may be an effective bone graft material for the regeneration of periodontal tissue in intrabony defects.

Effects of Scintillation Crystal Surface Treatments on Small Gamma Camera Imaging (섬광체 옆 표면처리가 소형 감마카메라 영상에 미치는 효과)

  • Kim, J. H.;Choi, Y.;Kim, J. Y.;Oh, C. H.;Kim, S. E.;Choe, Y. S.;Lee, K. H.;Joo, K. S.;Kim, B. T.
    • Journal of Biomedical Engineering Research
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    • v.20 no.6
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    • pp.515-521
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    • 1999
  • Scintillator crystal is an important part and detcrmines performance characteristics of the gamma camera. We investigated the offects of scintillation crystal surface treatment on gamma camera imaging. Nal(TI) and Csl(Tl) scintillators. 20 mm diameter and 10 mm thickness, applied with two different surface treatments, white and black reflcetors, were applied to Nal(Tl) and Csl(Ti). The optical properties of generated scintillation light were evaluated by Monte Carlo simulation method and by actual measurement using a position sensitive photomultiplier tube (PSPMT). We measured sensitivity, energy resolution and spatial resolution of gamma camera with the various scintillators coupled to a PSPMT. In the simulation. Nal(Tl)-white prosented the best sensitivity. In the measurements, the sensitivities and the intrinsic spatial resolutions of Nal(Tl)-white, Nal(Tl)-black. CsI(Tl)-white, CsI(Tl)-black were 2920, 2322, 1754, 1401 cps/$\mu$ci and 5.2, 4.5, 7.0, 6.3 mm FWHM. respectively. Their intrinsic energy resolutions were mesured 12.5, 23.5, 20.5, 33.3% FWHM at 140 keV Tc-99m. In this study, we investigated the offects of a side surface treatment of the scintillator on the gamma camera imaging. Simulation and measurement prescnted similat trends. Based on the results, we concluded that the surface of th NaI(Tl)seintillator must be treated by absorptive materials in order to develop the gamma camera having good spatial resolution.

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Development of Adaptive Spatial Filter to Improve Noise Characteristics of PET Images (PET 영상의 잡음개선을 위한 적응적 공간 필터 개발)

  • Woo, S. K.;Choi, Y.;Im, K. C.;Song, T. Y.;Jung, J. H.;Lee, K. H.;Kim, S. E.;Choe, Y. S.;Park, C. C.;Kim, B. T.
    • Journal of Biomedical Engineering Research
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    • v.23 no.3
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    • pp.253-261
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    • 2002
  • A spatially adaptive falter was formulated to imrove PET image qualify and the Performance of the filter was evaluated using simulation and phantom and human PET studies. In the proposed filter. if a pixel was identified as the edge Pixel, the Pixel value was Preserved. Otherwise a Pixel was replaced by the mean of the pixel values weighted by 2:7: 2. A Pixel was identified as the edge Pixel. if it satisfies the following conditions : the number of ADs (absolute difference between center and neighborhood pixels) which is smaller than THl (($pix_max{\times}0.1/log_2(NPM)$, NPM : mean of 6 neighborhood pixels excluding minimum and maximum) is 8-k and the number of ADs which is lager than TH2 ($NPM{\times}0.1$) is k. where k : 2, 3, …, 6. The results of this study demonstrate the superior performance of the Proposed titter compared to Gaussian fitter, weight median filter and subset averaged median filter. The proposed tittering method is simple but effective in increasing uniformity and contrast with minimal degradation of spatial resolution of PET images and thus. is expected to Provide improved diagnositc quality PET images .