• BMB Reports
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• 제40권5호
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• pp.625-635
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• 2007

The enzyme squalene synthase (EC 2.5.1.21) catalyzes a reductive dimerization of two farnesyl diphosphate (FPP) molecules into squalene, a key precursor for the sterol and triterpene biosynthesis. A full-length cDNA encoding squalene synthase (designated as TcSqS) was isolated from Taxus cuspidata, a kind of important medicinal plants producing potent anti-cancer drug, taxol. The full-length cDNA of TcSqS was 1765 bp and contained a 1230 bp open reading frame (ORF) encoding a polypeptide of 409 amino acids. Bioinformatic analysis revealed that the deduced TcSqS protein had high similarity with other plant squalene synthases and a predicted crystal structure similar to other class I isoprenoid biosynthetic enzymes. Southern blot analysis revealed that there was one copy of TcSqS gene in the genome of T. cuspidata. Semi-quantitative RT-PCR analysis and northern blotting analysis showed that TcSqS expressed constitutively in all tested tissues, with the highest expression in roots. The promoter region of TcSqS was also isolated by genomic walking and analysis showed that several cis-acting elements were present in the promoter region. The results of treatment experiments by different signaling components including methyl-jasmonate, salicylic acid and gibberellin revealed that the TcSqS expression level of treated cells had a prominent diversity to that of control, which was consistent with the prediction results of TcSqS promoter region in the PlantCARE database.

• 한국정보처리학회논문지
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• 제6권8호
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• pp.2179-2187
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• 1999

산발적인 지역의 사용자들에 대한 멀티미디어 이동 통신 서비스를 위하여 무선 회선을 이용한 ATM을 적용하는 것이 검토되고 있다. 무선 회선에서 효율적인 ATM 셀 전송으로 TDMA 방식을 적용하는 것이 유효하지만, TDMA와 ATM의 동기/비동기성의 차이로 인한 셀 지연변이를 보상하는 방법과 복수 사용자들에 대한 셀 스트림 다중방법이 요구된다. 본 논문에서는 ATM 셀 지연변이의 보상방식 중에서 많은 이점을 갖는 타임스탬프 방식을 사용하여 마르코프 변조 포이송(MMPP)으로 모델링한 VBR 트래픽 입력인 경우에 대하여 지연변이에 따른 셀 제어시간(Tc)를 최적화한다. 또한, 멀티미디어 이동 통신 서비스를 제공하는 무선 ATM에서의 셀 스트림 재생시 복수 사용자들의 셀 스트림 다중시에 발생하는 셀 지연 변이(CDV)에 적용한 셀 스트림 다중방식을 제안하고, 서비스 전송 특성을 시뮬레이션을 수행하였다. 본 방식의 적용에 따라 CDV 분포폭은 약 1.2Tc 정도로 억제가 가능하여 전체적인 셀 지연변이가 감소되었으며, 셀 스트림을 재생하고, 동시에 복수 셀 스트림을 용이하게 다중화가 가능함을 알 수 있었다.

• 한국정보처리학회논문지
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• 제3권3호
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• pp.512-522
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• 1996

산발적인 지역의 가입자들에 대한 광대역 ISDN(B-ISDN) 서비스를 위하여 위성 회선을 적용하는 것이 검토 되고 있다. 위성 회선의 효율적인 ATM 셀 전송으로 TDMA 방식을 적용하는 것이 유효하지만, TDMA와 ATM의 동기/비동기성의 차이로 인한 셀 지연 변이를 보상하는 방법이 필요하게 된다. 본 논문에서는 지금까지 제안되어 왔던 셀 지연 변이의 보상 방식 중에서 많은 이점을 갖는 셀 계수 방식의 지연 특성을 응용 함으로써 트래픽이 포아송 입력, 마르코프 변조 포아송 과정 입력인 경우에 대하여 셀 제어 시간(Tc)의 적화를 유도하였다. 그리고 셀 클럼핑 현상을 억제하기 위하여 이산 타임 스탬프 방식을 제안 하여 ATM 전송에 따른 CDV 분포의요구 품질의 범위를 해석 하고 검증하였다. 본 방식의 적용에 따라 CDV 분포폭은 약1.2$\times$Tc 정도로 억제가 가능 하여이산 타임 스탬스방식에 따른 전체적인 셀 지연 변이가 감소됨을 알 수 있었다.

• KSBB Journal
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• 제13권5호
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• pp.561-568
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• 1998

Two cometabolic trichloroethylene (TC) degraders, Pseudomonas putida F1 and Burkholderia (Pseudomonas) cepacia G4, were found to catabolize phenol, benzene, toluene, and ethylbenzene as carbon and energy sources. Resting cells of P. putida F1 and B. cepacia G4 grown in the presence of toluene and phenol, respectively, were able to degrade not only benzene, toluene and ethylenzene but also TCE and p-xylene. However, these two strains grown in the absence of toluene or phenol did not degrade TCE and p-xylene. Therefore, it was tentatively concluded that cometabolic degradation of TC and p-xylene was mediated by toluene dioxygenase (P. putida F1) or toluene-2-monooxygenase (B. cepacia G4). Maximal degradation rates of BTEX and TCE by toluene- and phenol-induced resting cells of P. putida F1 and B. cepacia G4 were appeared to be 4-530 nmol/(min$.$mg cell protein) when a single compound was solely served as a target substrate. In case of double substrates, the benzene degradation rate by P. putida F1 in the presence of toluene was decreased up to one seventh of that for the single substrate. TCE degradation rate was also linearly decreased as toluene concentration increased. On the other hand, toluene degradation rate was enhanced by benzene and TCE. For B. cepacia G4, degradation rates of TCE and toluene increased 4 times in the presence of 50 ${\mu}$M phenol. From these results, it was concluded that a degradation rate of a compound in the presence of another cosubstrate(s) could not be predicted by simply generalizing antagonistic or synergistic interactions between substrates.

• Journal of Periodontal and Implant Science
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• 제32권1호
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• pp.13-23
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• 2002

The ultimate goal of periodontal therapy is directed to arresting the progression of the disease, and regenerating the fibrous attachment. In order to achieve such treatment aim, the plaque and calculus must be eliminated and the physiological conditions of the root surface must be changed to facilitate the attachment and migration of the new fibroblasts, The method of changing the proper root surface conditions to promote the healing of periodontal tissue involves mechanical procedures, such as scaling and root planing, and chemical procedures such as tetracycline-HCl. However, the formation of a long junctional epithelium was most frequently observed type of healing. Thus, the aim of this study was to examine in vitro the influence of surface conditioning of dentin by TC-HCl on human gingival epithelial cell attachment. Human gingival epithelial cells were obtained from healthy retromolar pad area(under the age 23 years). Seventy two teeth extracted from severe periodontitis were used as study material. To evaluate the epithelial cell attachment to dentin, the prepared specimen was divided to four groups. For the control group, only scaling and root planing were carried out, and for the test group, 1 to 3, the concentration of the TC-HCl was 50, 125 and 250mg/ml respectively. After cell cultivation time of 1-, 3-. 24 hour, for the indirect quantitative assessment of gingival epithelial cell attached to dentin sample, the absorbance of epithelial cell unattached to dentin was measured. The results were as follows; 1. There was no statistically significant difference between scaling and root planing group and TC-HCl 50mg/ml 125mg/ml and 250mg/ml group about absorbance of unattached epithelial cell to dentin sample(p>0.5). 2. As time passes, the absorbance of unattached gingival epithelial cell to dentin sample was decreased statistically significant(p<0.05). 3. There was no statistically significant difference among the TC-HCl group(p>0.05) We concluded that there was similar effect on gingival epithelial cell attachment between TC-HCl conditioning on root surface and only scaling and root planing treatment

• 대한핵의학회지
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• 제29권1호
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• pp.125-132
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• 1995

The purpose of this study was to establish mononuclear cell cultures such as lymphocytes or buffy coat for the biological dosimetry of in vitro Irradiation of the radionuclide Tc-99m in order to exclude the effect of residual doses seen in the cultures of whole blood. Biological do simetry of Tc-99m on cultured mononuclear cells at doses ranging from 0.05 to 6.00 Gy, by scoring unstable chromosomal aberrations(Ydr) observed in cultured lymphocytes, were performed using peripheral venous blood of healthy normal person. The results showed that; (1) In vitro irradiation of radioisotope in separated lymphocyte or buffy coat showed trace amount of residual doses of isotope after washing. Residual doses of isotopes are increased in proportion to exposed time and irradiated dose without difference between I-131 and Tc-99m. (2) We obtained these linear-quadratic dose response equations in lymphocyte and buffy coat culture after in vitro irradiation of Tc-99m, respectively (Ydr = 0.001949 $D^2$ +0.006279D + 0.000185; Ydr= 0.002531 $D^2$-0.003274 D+0.003488). In conclusion, the linear quadratic dose-response equation from in vitro irradiation of Tc-99m with lymphocyte and buffy coat culture was thought to be useful for assessing Tc-99m induced biological effects. And mono-nuclear cell cultures seem to be the most appropriate experimental model for the assessment of biological dosimetry of internal irradiation of radionuclides.

• 대한의생명과학회지
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• 제15권1호
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• pp.55-60
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• 2009

Insect cells such as Spodoptera frugiperda Clone 9 (Sf9) cells are widely chosen as the host for heterologous expression of a mammalian sugar transport protein using the baculovirus expression system. Characterization of the expressed protein is expected to include assay of its function, including its ability to transport sugars and to bind inhibitory ligands such as cytochalasin B. It is therefore very important first to establish the transport characteristics and other properties of the endogenous sugar transport proteins of the host insect cells. However, very little is known of the transport characteristics of Sf9 cells, although their ability to grow on TC-100 medium strongly suggested the presence of endogenous glucose transport system. In order to investigate the substrate and inhibitor recognition properties of the Sf9 cell transporter, the ability of pentoses to inhibit 2-deoxy-D-glucose (2dGlc) transport was investigated by measuring inhibition constants $(K_i)$. To determine the time period over which of sugar into the Sf cells was linear, the uptake of 2dGlc 0.1mM extracellular concentration was measured over periods ranging from 30 seconds to 30 minutes. The uptake was linear for at least 2 minutes at the concentration, implying that uptake made over a 1 minute time course would reflect initial rates of the sugar uptake. The data have also revealed the existence of a saturable transport system for pentose uptake by the insect cells. The transport was inhibited by D-xylose and D-ribose, although not as effective as hexoses. However, L-xylose had a little effect on 2dGlc transport in the Sf9 cells, indicating that the transport is stereoselective. Unlike the human erythrocyte-type glucose transport system, D-ribose had a somewhat greater apparent affinity for the Sf9 cell transporter than D-xylose. It is therefore concluded that Sf9 cells contain an endogenous sugar transport activity that in some aspects resembled the human erythrocyte-type counterpart, although the Sf9 and human transport systems do differ in their affinity for cytochalasin B.

• 대한핵의학회지
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• 제23권2호
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• pp.219-223
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• 1989

Inflammation scan using radiolabelled leukocytes has high sensitivity and specificity. Several methods for labelling leukocytes have been evaluated using P-32 diisopropyl fluorophosphate (DFP-32), H-3 thymidine, Cr-51 chromate, Ga-67 citrate and Tc-99m-sulfur colloid. In-111-oxine has proved so far to be the most reliable agent for labelling leukocytes. In-111-oxine is, however, expensive, not easily available when needed, and its radiation dose to leukocytes is relatively high. Moreover, resolution of the resultant image is relatively poor. Tc-99m is still the agent of choice because of, as compared with the indium, its favorable physical characteristics, lower cost and availability. Now the technique for labelling the leukocytes with technetium is successfully obtained using the lipophilic HAPAO with higher efficiency for granulocytes than for other cells. With this technique it is possible to label leukocytes in plasma to improve the viability of the leukocytes. Inflammation scan using Tc-99m-HMPAO has been evaluated in several laboratories, and difference in methods for separation and labelling accounts for difference in efficiency, viability and biodistribution of the labelled leukocytes. We performed inflammation scan using leukocytes labelled with Tc-99m-HMPAO in three dogs 24 hours after inoculation of live E. Coli and A. Aureus in their right abdominal wall. We separated mixed leukocytes by simple sedimentation using 6% hetastarch (HES) and labelled the leukocytes with Tc-99m-HMPAO in 20% cell free plama diluted with phosphate buffer solution(Fig. 1). Uptake was high in the liver and spleen but is was minimal in the lungs on whole body scan. Kidneys and intestine showed minimal activity although it was high in the urinary bladder(Fig. 2). Uptake of labelled leukocytes in the inflammation site was do(mite on 2 hour-postinjection scan and abscess was clearly delineated on 24 hour-delayed scan with high target-to-nontarget ratio(Fig. 3, 4). Inflammation scan using mixed leukocytes labelled with Tc-99m-HMPAO is very sensitive and specific in early detection of inflammation.

• 핵의학기술
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• 제14권1호
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• pp.35-39
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• 2010

감시림프절 검사에는 $^{99m}Tc$-ASC, $^{99m}Tc$-Tin colloid, $^{99m}Tc$-HSA 등이 사용되었으나 공급 제한으로 간 스캔에 사용되고 있는 $^{99m}Tc$-phytate를 대체 방사성의약품으로 사용하고 있다. 이에 저자들은 phytate의 입자 크기 분포를 알아보고 200 nm 필터 사용유무에 따른 영상을 획득 분석하여 감시림프절 검사 시 필터 사용의 적절성을 판단하고자 하였다. Photal사의 ELS-8000 장비를 이용하여 phytate kit의 입자크기 분포를 측정 하였다. 동물 실험은 생후 12~15주령, 체중 250~300 g 암컷 백서를 사용하였으며, 전신 마취 후 앙와 위로 고정시켜 영상을 획득하였다. 비교군으로 여과시키지 않은 $^{99m}Tc$-phytate를 실험군으로는 200 nm 필터를 이용하여 여과시킨 $^{99m}Tc$-phytate를 백서의 발가락 사이에 피하주사 하였다. 6마리의 백서를 3~4일 간격으로 비교군과 대조군으로 그룹을 나누어 2회씩 실시 하였으며, 주사 후 1시간 지연영상을 얻은 후 서혜부에 관심영역을 설정하였으며 비정상적인 집적이 없는 부위를 선별하여 픽셀당 카운트를 얻었다. 세포 실험은 RAW 264.7 세포를 이용하여 비교군과 실험군에 $^{99m}Tc$-phytate를 넣어주고 30분 동안 배양시켜 감마카운터를 이용하여 각각의 분 당 카운트 값을 측정하였다. 이 값들은 SPSS 12.0 프로그램을 이용하여 통계 검정하였다. Phytate를 생리식염수에 용해시킨 후 측정 결과 평균 직경은 130~650 nm가 $85{\pm}5%$ 정도였으나 200 nm 필터를 이용하여 여과시킨 후의 입자 크기는 용매 내 phytate 입자의 농도가 너무 낮아 측정할 수 없었다. 획득한 영상을 분석한 결과 비교군에서는 $60.2{\pm}4.88$ count/pixel, 실험군에서는 $58.3{\pm}5.97$ count/pixel 값을 보였다. 세포 실험에서 실험군의 카운트 값은 $17,152{\pm}1,165$ cpm이고 비교군은 $16,908{\pm}1,075$ cpm을 보였다. 동물 실험과 마찬가지로 세포 실험에서도 통계적 차이는 보이지 않았다. 이번 실험을 통하여 phytate의 사이즈가 매우 분산적이며 균일하지 못 하다는 사실을 확인하였으며, 동물 실험과 세포실험을 통한 필터 사용의 적절성을 판단한 결과 phytate를 이용한 감시림프절 검사에서 필터를 사용하는 방법은 결과에 큰 영향을 미치지 않는다는 것을 확인하였다.

• Radiation Oncology Journal
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• 제25권3호
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• pp.185-191
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• 2007

목적 : 가시오갈피 뿌리 추출물(Extract of Eleuthrococcus senticosus Maxim root, ESMR)의 면역세포의 활성에 미치는 효과와 마우스 종양세포에서 세포독성 및 방사선 병용효과를 알기위해서 본 연구를 시행하였다. 대상 및 방법: 가시오갈피 뿌리(250 g)를 80% 메탄올로 추출한 다음, 감압농축 및 동결과정을 통해 분획을 얻었다. 세포 주는 섬유육종세포를 이용하였으며, 방사선조사는 저 에너지(250 KV)용 방사선 기기를 사용 하였다. 가시오갈피 추출물의 면역세포에 미치는 활성효과를 알아보기 위해서 마우스 비장 및 흉선세포의 생존율 측정과 비장 및 흉선림프구 아집단을 측정하였다. 세포독성 정도를 알아보기 위하여 clonogenic assay법을 이용하였다. 방사선조사 단독군의 경우 2, 4 및 6 Gy를 마우스 섬유육종세포(FSa II)에 조사하였으며, 가시오갈피와 방사선 병용군의 경우 0.2 mg/ml 농도의 가시오갈피를 방사선조사 1시간 전에 마우스 섬유육종세포에 접촉시켰다. 결 과: 가시오갈피 /뿌리 추출물 각각 10 ${\mu}g/ml$과 $100{\mu}g/ml$을 첨가한 마우스 비장 및 흉선세포 생존율은 다음과 같았다. 마우스 비장세포는 각각 $8.8{\pm}1.7%$와 $37.7{\pm}2.1%$로 증가하였으며, 흉선세포에서는 100 ${\mu}g/ml$에서 $16.7{\pm}1.6%$로 증가하였다. ESMR을 경구 투여한 마우스 비장 및 흉선림프구는 다음과 같았다. 마우스 비장세포의 B cell은 41.5 $({\pm}1.3)$에서 46.0 $({\pm}1.5)%$로, T cell은 16.9 $({\pm}0.9)$에서 22.3 $({\pm}1.2)%$로 증가하였으며, 특히 Helper T (Th) cell은 10.4 10.4 $({\pm}0.7)$에서 14.2 $({\pm}0.8)%$로, Cytotoxic T (Tc) cell은 3.6 $({\pm}0.4)$에서 5.6 $({\pm}0.3)%$로 증가하였다. 그러나 마우스 흉선세포에서 T림프구의 증가율은 거의 보이지 않았다. 마우스 섬유육종 세포에서 ESMR의 세포독성을 나타내는 생존분율은 0.2, 2.0 mg/m에서 각각 $0.36{\pm}0.02$, $0.05{\pm}0.02$으로 나타났다. 마우스 섬유육종 세포를 접종한 마우스에 ESMR 0.2 mg/ml 농도를 1시간 동안 접촉시킨 다음, 방사선조사량 2, 4 및 6 Gy를 조사한 후 얻은 생존분물은 0.39, 0.22 및 0.06이었으며, 방사선조사 단독군의 경우는 0.76, 0.47 및 0.37로 나타났다. 그리고 두 군간의 차이는 통계학적 의미를 나타냈다(p<0.05). 결 론: 가시오갈피 뿌리 추출물은 in vitro 실험에서 마우스 비장세포의 생존율을 증가시켰다. In vivo 실험에서는 마우스 비장 내 B세포 및 T세포를 증가시켰으나, 마우스 흉선세포에 대해서는 증가를 보이지 않았다. ESMR은 마우스 섬유육종 세포에 대해 강한 세포독성 효과를 나타냈다. FSa II를 접종한 마우스에서 방사선 단독 보다 ESMR을 병용한 경우 방사선에 의한 세포손상이 약 50% 이상 증가되었으며, 두 군간의 차이는 통계학적 의미를 나타냈다(p<0.05).