• Title/Summary/Keyword: p53/sup +/-/

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Fed-batch Fermentations of Recombinant Escherichia coli to Produce Bacillus macerans CGTase

  • Park, Yong-Cheol;Kim, Chang-Sup;Kim, Chung-Im;Choi, Kyu-Hwan;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.7 no.5
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    • pp.323-328
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    • 1997
  • The recombinant Escherichia coli BL21(DE3)pLysE : pTCGT1 was grown to overproduce Bacillus macerans cyclodextrin glucanotransferase (CGTase) able to synthesize ${\alpha}$-cyclodextrin (CD) with a selectivity of 67%. A number of batch fermentations were performed to test the possibility of using lactose as an inducer of the E. coli T7 promoter system. A mixture of isopropyl ${\beta}$-D-thiogalactoside (IPTG) and lactose (1 : 1) gave a maximum CGTase activity of 2.4 U/ml, which was higher than the value obtained with induction by IPTG alone. Fed-batch fermentations involving a glucose-controlled growth period followed by a gene-expression phase with mixtures of IPTG and lactose were employed to achieve high cell density and thereby increase total CGTase activity. Optimized fed-batch fermentation using the modified inducer (IPTG : lactose=1 : 3) and 100 g/l yeast extract solution in the gene-expression phase resulted in a maximum CGTase activity of 62.9 U/ml and a final cell mass of 53.5 g/l, corresponding to a 31-fold increase in CGTase activity and a 29-fold increase in cell mass compared with the control batch fermentation.

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Overexpression of Anti-apoptotic Molecules and Sax Translocation to Mitochondria by Pharbitis Nil Extracts in AGS

  • Ko Seong-Gyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.6
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    • pp.1843-1849
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    • 2004
  • Conventional medicines have usually sorted to a number of treatments such asoperation, radiotherapy, and chemotherapy. The existing anti-cancer agents, designed to eradicate cancer cells, have strong toxicities, also with leading to harmful side effects. Recently, a number of researches on natural products have been actively carried out in efforts to develop new treatments that can decrease side effects or increase anti-cancer effects. We performed this study to understand the molecular basis underlying the antitumor effects of Pharbitis nil, and Plantago asiatica, which have been used for herbal medicinal treatments against cancers in East Asia. We analyzed the effects of these medicinal herbs on proliferation and on expression of cell growth/apoptosis related molecules, with using an AGS gastric cancer cell line. The treatment of Pharbitis nil dramatically reduced cell viabilities in a dose and time-dependent manner, but Plantago asiatica didn't. FACS analysis and Annexin V staining assay also showed that Pharbitis nil induce apoptotic cell death of AGS. Expression analyses via RT-PCR and Western blots revealed that Pharbitis nil didn't increase expression of the p53 and its downstream effector p21/sup wafl/, and that the both increased expression of apoptosis related Sax and cleavage of active caspase-3 protein. We also confirmed the translocation of Sax to mitochondria. Collectively, our data demonstrate that Pharbitis nilinduce growth inhibition and apoptosis of human gastric cancer cells, and these effects are correlated with down- and up-regulation of growth-regulating apoptotic and tumor suppressor genes, respectively.

Antiproliferative Effect of RST Associated with the Inhibition of Cyclooxygenase-2 Expression and Prostaglandin E2 Release in Human Lung Carcinoma Cells (산두근 추출물이 인체폐암세포의 COX-2 발현 및 PGE2 생성에 미치는 영향)

  • Kim, Kang-Tae;Eom, Hyun-Sup;Chi, Gyoo-Yong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.4
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    • pp.907-915
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    • 2007
  • In this study the effect of water extract of Sophora tonkinensis Gapnep (RST) was investigated on the growth of human lung carcinoma A549 cells. Exposure of A549 cells to RST resulted in the growth inhibition in a dose-dependent manner as measured by MTT assay. The antiproliferative effect by RST treatment in A549 cells was associated with morphological changes such as membrane shrinking and cell rounding up. RST treatment did not induce the cell cycle arrest and the levels of tumor suppressor p53 as well as cyclin-dependent kinase inhibitor p21(WAF1/CIP1). It was found that RST treatment decreased the levels of cyclooxygenase (COX) -2 mRNA and protein expression without significant changes in the expression of COX-1 and inducible nitric oxide synthase (iNOS), which was correlated with a decrease in prostaglandin E2 (PGE2) synthesis. RST treatment also slightly inhibited the levels of human telomerase reverse transcriptase (hTERT) mRNA and protein expression, and the activity of telomerase. Taken together, these findings suggested that RST-induced inhibition of human lung carcinoma A549 cell growth was aoosciated with the inhibition of COX-2 expression and PGE2 production. These results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of RST.

In Vivo Image and Biodistribution of $^{123}I$-15-(p-iodophenyl)-3-R, S-methylpentadecanoic acid (BMIPP) in Liposarcoma Bearing Nude Mice (지방육종형성 동물모델에서 $^{123}I$-15-(p-iodophenyl)-3-R, S-methylpentadecanoic acid (BMIPP)의 생체분포와 생체영상)

  • Lee, Tae-Sup;Suh, Yong-Sup;Choi, Chang-Woon;Woo, Kwang-Sun;Chung, Wee-Sup;Lim, Soo-Jung;Lim, Sang-Moo;Awh, Ok-Doo
    • The Korean Journal of Nuclear Medicine
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    • v.35 no.5
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    • pp.324-333
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    • 2001
  • Purpose: $^{123}I$-labeled fatty acids have been used in the evaluation of regional myocardial energy metabolism. This study aimed to evaluate the usefulness of $^{123}I$-BMIPP as a liposarcoma-imaging agent. Materials and Methods: We compared in vitro uptakes between liposarcoma(SW872) and glioma(9L) cell lines, and examined biodistribution and in vivo images of $^{123}I$-BMIPP in liposarcoma-bearing nude mice. Cold-BMIPP was labeled with $^{123}I\;using\;Cu^{2+}$ as catalyst. After purification by Sep-pak, radiochemical purity was determined by TLC. We compared cellular uptake between glioma and liposarcoma after incubation of 5, 10, 15, 30, 60, 120, and 180 mins with culture medium containing $^{123}I$-BMIPP. The difference in biodistribution was determined between non-feeding (water only) group for 18 hr and feeding group in normal mice (n=6/group) at 0.5, 2, and 24 hr. In liposarcoma-hearing nude mice model, liposarcoma, SW872, ceil lines were injected subcutaneously into the felt thigh of nude mice. The biodistribution of $^{123}I$-BMIPP was evaluated at 0.5, 2, and 24 hr (n:5 / group) and in vivo Image of $^{123}I$-BMIPP was obtained with gamma camera at 2 and 24 hr in liposarcoma-hearing nude mice. Results: Radiolabeling yield and radiochemical purity were 95% and above 99%, respectively. SW872 cell line showed more increased uptake than 9L with 1.5 times at 180 mins. The clearance of $^{123}I$-BMIPP in various tissues was more delayed in the non-feeding group than in the feeding group, especially at delayed time (24 hr) in normal mice, and the major excreting organ was the gastrointestinal tract. In liposarcoma-bearing nude mice, tumor/blood ratio of $^{123}I$-BMIPP was 0.94, 0.75, and 1.38 and tumor/muscle ratio was 0.66, 1.53, and 1.11 at 0.5, 2, and 24hr, respectively. $^{123}I$-BMIPP was selectively localized in liposarcoma at 24 hr image. Conclusions: These results suggest that $^{123}I$-BMIPP can be used as a liposarcoma-imaging agent.

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Evaluation of the Usefulness of Virtual Reality Equipment for Relieving Patients' Anxiety during Whole-Body Bone Scan (전신 뼈 검사 환자의 불안감 해소를 위한 가상현실 장비의 유용성 평가)

  • Kim, Hae-Rin;Kim, Jung-Yul;Lee, Seung-Jae;Baek, Song-Ee;Kim, Jin-Gu;Kim, Ga-Yoon;Nam-Koong, Hyuk;Kang, Chun-Goo;Kim, Jae-Sam
    • The Korean Journal of Nuclear Medicine Technology
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    • v.26 no.1
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    • pp.27-32
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    • 2022
  • Purpose When performing a whole-body bone scan, many patients are experiencing psychological difficulties due to the close distance to the detector. Recently, in the medical field, there is a report that using virtual reality (VR) equipment can give pain relief to pediatric patients with weak concentration or patients receiving severe treatment through a distraction method. Therefore, in this paper, VR equipment was used to provide psychological stability to patients during nuclear medicine tests, and it is intended to evaluate whether it can be used in clinical practice. Materials and Methods As VR equipment, ALLIP Z6 VR (ALLIP, Korea) was used and the experiment was conducted after connecting to a mobile phone. The subjects were 30 patients who underwent whole-body bone examination from September 1, 2021 to September 30, 2021. After intravenous injection of 99mTc-HDP, 3 to 6 hours later, VR equipment was put on and whole body images were obtained. After the test, a survey was conducted, and a Likert scale of 5 points was used for psychological anxiety and satisfaction with VR equipment. Hypothesis verification and reliability of the survey were analyzed using SPSS Statistics 25 (IBM, Corp., Armonk, NY, USA). Results Anxiety about the existing whole-body bone test was 3.03±1.53, whereas that of anxiety after wearing VR equipment was 2.0±1.21, indicating that anxiety decreased to 34%. When regression analysis of the effect of the patient's concentration on VR equipment on anxiety about the test, the B value was 0.750 (P<0.01) and the t value was 6.181 (P<0.01). decreased and showed an influence of 75%. In addition, overall satisfaction with VR equipment was 3.76±1.28, and the intention to reuse was 66%. The Cronbach α value of the reliability coefficient of the questionnaire was 0.901. Conclusion When using VR equipment, patients' attention was dispersed, anxiety was reduced, and psychological stability was found. In the future, as VR equipment technology develops, it is thought that if the equipment can be miniaturized and the resolution of VR content images is increased, it can be used in various clinical settings if it provides more realistic stability to the patient.

Detection of Contralateral Breast Cancer Using Diffusion-Weighted Magnetic Resonance Imaging in Women with Newly Diagnosed Breast Cancer: Comparison with Combined Mammography and Whole-Breast Ultrasound

  • Su Min Ha;Jung Min Chang;Su Hyun Lee;Eun Sil Kim;Soo-Yeon Kim;Yeon Soo Kim;Nariya Cho;Woo Kyung Moon
    • Korean Journal of Radiology
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    • v.22 no.6
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    • pp.867-879
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    • 2021
  • Objective: To compare the screening performance of diffusion-weighted (DW) MRI and combined mammography and ultrasound (US) in detecting clinically occult contralateral breast cancer in women with newly diagnosed breast cancer. Materials and Methods: Between January 2017 and July 2018, 1148 women (mean age ± standard deviation, 53.2 ± 10.8 years) with unilateral breast cancer and no clinical abnormalities in the contralateral breast underwent 3T MRI, digital mammography, and radiologist-performed whole-breast US. In this retrospective study, three radiologists independently and blindly reviewed all DW MR images (b = 1000 s/mm2 and apparent diffusion coefficient map) of the contralateral breast and assigned a Breast Imaging Reporting and Data System category. For combined mammography and US evaluation, prospectively assessed results were used. Using histopathology or 1-year follow-up as the reference standard, cancer detection rate and the patient percentage with cancers detected among all women recommended for tissue diagnosis (positive predictive value; PPV2) were compared. Results: Of the 30 cases of clinically occult contralateral cancers (13 invasive and 17 ductal carcinoma in situ [DCIS]), DW MRI detected 23 (76.7%) cases (11 invasive and 12 DCIS), whereas combined mammography and US detected 12 (40.0%, five invasive and seven DCIS) cases. All cancers detected by combined mammography and US, except two DCIS cases, were detected by DW MRI. The cancer detection rate of DW MRI (2.0%; 95% confidence interval [CI]: 1.3%, 3.0%) was higher than that of combined mammography and US (1.0%; 95% CI: 0.5%, 1.8%; p = 0.009). DW MRI showed higher PPV2 (42.1%; 95% CI: 26.3%, 59.2%) than combined mammography and US (18.5%; 95% CI: 9.9%, 30.0%; p = 0.001). Conclusion: In women with newly diagnosed breast cancer, DW MRI detected significantly more contralateral breast cancers with fewer biopsy recommendations than combined mammography and US.

The cumulative survival rate of dental implants with micro-threads: a long-term retrospective study

  • Dong-Hui Nam;Pil-Jong Kim;Ki-Tae Koo;Yang-Jo Seol;Yong-Moo Lee;Young Ku;In-Chul Rhyu;Sungtae Kim;Young-Dan Cho
    • Journal of Periodontal and Implant Science
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    • v.54 no.1
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    • pp.53-62
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    • 2024
  • Purpose: This study aimed to evaluate the long-term cumulative survival rate (CSR) of dental implants with micro-threads in the neck over a 10-year follow-up period and to examine the factors influencing the survival rate of dental implants. Methods: This retrospective study was based on radiographic and dental records. In total, 151 patients received 490 Oneplant® dental implants with an implant neck micro-thread design during 2006-2010 in the Department of Periodontology of Seoul National University Dental Hospital. Implant survival was evaluated using Kaplan-Meier analysis. Cox proportional hazard regression analysis was used to identify the factors influencing implant failure. Results: Ten out of 490 implants (2.04%) failed due to fixture fracture. The CSR of the implants was 97.9%, and no significant difference was observed in the CSR between external-and internal-implant types (98.2% and 97.6%, respectively, P=0.670). In Cox regression analysis, 2-stage surgery significantly increased the risk of implant failure (hazard ratio: 4.769, P=0.039). There were no significant differences in influencing factors, including sex, age, implant diameter, length, fixture type, location, surgical procedure, bone grafting, and restoration type. Conclusions: Within the limitations of this retrospective study, the micro-thread design of the implant neck was found to be favorable for implant survival, with stable clinical outcomes.

No benefit of hypomethylating agents compared to supportive care for higher risk myelodysplastic syndrome

  • Sohn, Sang Kyun;Moon, Joon Ho;Lee, In Hee;Ahn, Jae Sook;Kim, Hyeoung Joon;Chung, Joo Seop;Shin, Ho Jin;Park, Sung Woo;Lee, Won Sik;Lee, Sang Min;Kim, Hawk;Lee, Ho Sup;Kim, Yang Soo;Cho, Yoon Young;Bae, Sung Hwa;Lee, Ji Hyun;Kim, Sung Hyun;Song, Ik Chan;Kwon, Ji Hyun;Lee, Yoo Jin
    • The Korean journal of internal medicine
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    • v.33 no.6
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    • pp.1194-1202
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    • 2018
  • Background/Aims: This study evaluated the role of hypomethylating agents (HMA) compared to best supportive care (BSC) for patients with high or very-high (H/VH) risk myelodysplastic syndrome (MDS) according to the Revised International Prognostic Scoring System. Methods: A total of 279 H/VH risk MDS patients registered in the Korean MDS Working Party database were retrospectively analyzed. Results: HMA therapy was administered to 205 patients (73.5%), including 31 patients (11.1%) who then received allogeneic hematopoietic cell transplantation (allo-HCT), while 74 patients (26.5%) received BSC or allo-HCT without HMA. The 3-year overall survival (OS) rates were $53.1%{\pm}10.7%$ for allo-HCT with HMA, $75%{\pm}21.7%$ for allo-HCT without HMA, $17.3%{\pm}3.6%$ for HMA, and $20.8%{\pm}6.9%$ for BSC groups (p < 0.001). In the multivariate analysis, only allo-HCT was related with favorable OS (hazard ratio [HR], 0.356; p = 0.002), while very poor cytogenetic risk (HR, 5.696; p = 0.042), age ${\geq}65years$ (HR, 1.578; p = 0.022), Eastern Cooperative Oncology Group performance status (ECOG PS) 2 to 4 (HR, 2.837; p < 0.001), and transformation to acute myeloid leukemia (AML) (HR, 1.901; p = 0.001) all had an adverse effect on OS. Conclusions: For the H/VH risk group, very poor cytogenetic risk, age ${\geq}65years$, ECOG PS 2 to 4, and AML transformation were poor prognostic factors. HMA showed no benefit in terms of OS when compared to BSC. Allo-HCT was the only factor predicting a favorable long-term outcome. The use of HMA therapy did not seem to have an adverse effect on the transplantation outcomes. However, the conclusion of this study should be carefully interpreted and proven by large scale research in the future.

Development of test method for the evaluation of pesticide acute toxicity using earthworm(Lumbricus rubellus) (붉은지렁이 (Lumbricus rubellus)를 이용한 농약의 급성독성 시험법 개발)

  • Park, Yeon-Ki;Park, Kyeong-Hoon;Kim, Byung-Seok;Kyung, Kee-Sung;Shin, Jin-Sup;Oh, Byung-Youl
    • The Korean Journal of Pesticide Science
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    • v.4 no.4
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    • pp.56-60
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    • 2000
  • A study was performed to determine the maintenance conditions of the earthworm, Lumbricus rubellus, for the acute toxicity test. To fine out climatic and soil conditions, the earthworms were maintained in artificial soil consisting of sand, clay mineral and peat at different levels of conditions for 14 days. Lumbricus rubellus led to an increase of biomass at temperature $22{\pm}2^{\circ}C$, soil pH $7.0{\pm}1$ and moisture 40%. And four fungicides were tested for acute toxicities to Lumbricus rubellus, according to the optimum condition. The test earthworms were exposed to each pesticide with various concentration gradients. After 14 days, the number of surviving earthworms and their weight alteration during the test period was determined. The 14-day $LC_{50}$ values for the Lumbricus rubellus, of carbendazim, benomyl, thiophanate-methyl and thiabendazole were determined to be 59, 53, 64 and 36 mg/ kg soil dry weight, respectively.

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Isolation of Anticarcinogenic Isoflavone-conjugated Glycoproteins from a Submerged Liquid Culture of Agaricus blazei Mycelia by the Autolysis Process (신령버섯균사체 액체배양물의 자가분해에 의한 항암성 isoflavone-conjugated glycoprotein 분리)

  • Kim, So Young;Kim, Young Suk;Jang, Joung Soon;Kim, Boh Hyun;Rakib, Abdur Md.;Kim, Gon Sup;Kim, Jeong Ok;Ha, Yeong Lae
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1316-1324
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    • 2014
  • Most beta-glucans obtained from various fruit bodies of mushrooms and mushroom mycelial cultures have high-molecular weight glycoproteins, conjugated with beta-glucans. We report that isoflavone-conjugated glycolproteins (designated as gluvone) were isolated and exhibited stronger anticarcinogenic activities. Agaricus blazei mycelia (ABM) was cultured in a liquid medium containing soybean flakes for 14 days. The liquid culture was autolyzed by incubating at $53^{\circ}C$ (pH 5.5) for 3 h. A crude glycoprotein (CGP) fraction with a cytotoxic effect on a mouse ascite cancer cell line (S-180) and a human breast cancer cell line (MCF-7) was isolated from the autolyzed ABM cultures by 80% ethanol treatment. Gluvone was isolated from the CGP with Sephadex G-75 column chromatography. It exhibited a stronger anticancer effect than CGP against the S-180 cell-induced female ICR mouse ascites carcinogenesis. Gluvone with 9,400 daltons was identified as a glycoprotein conjugated with isoflavone. According to HPLC and GC analysis, in conjunction with $^1H$-NMR spectral analysis, it contained 60% carbohydrates (glucose, fructose, and ribose), 31% protein, and 2% isoflavone (daidzein and genistein), which is a novel material. These results indicate that a strong anticarcinogenic gluvone was isolated from the autolyzed product of a submerged liquid culture of ABM, suggesting that autolysis could be a useful tool to produce antitumor agents.