• 생명과학회지
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• 제21권7호
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• pp.961-968
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• 2011

지질과산화로부터 생성된 aldehyde 중 4-hydroxynonenal (HNE)는 산화적 손상과 관련된 다량의 arachidonic acids, linoleic acids 등으로부터 생성될 수 있다. 그러므로 HNE는 산화적 스트레스와 관련된 세포사에서 중요한 매개인자로 작용을 할 수가 있을 것이다. 본 연구는 HNE가 세포사를 유발할 것이라 가정하고, 먼저 흰쥐 전립선 유래 내피세포인 YPEN-1 세포에서 세포독성을 측정하였다. 세포생장 저해능력은 HNE를 $5\sim15\;{\mu}M$ 농도로 처리하여 형태적 변화와 MTT assay를 통하여 결과를 관찰하였다. 그 결과 HNE가 이 세포에서 핵형의 변화와 세포사를 유발시키는 것을 각각의 실험을 통해 확인이 되었다. 또한 이 사실을 단백질의 변화를 통하여 확인을 할 수가 있었다. HNE를 24시간 처리한 세포에서 poly(ADP-ribose) polymerase 단백질 분절이 매개되었고 Bax의 발현량이 증가하였다. 또한 세포내의 활성 산소종들을 발생시켰다. 이에 생성된 활성 산소종과 peroxynitrite가 세포사와 관련이 있는가를 밝히기 위하여 이들의 포식자들인 N-acetylcysteine과 penicillamine을 본 연구에서 사용하였다. 이들 포식자들에 의해 HNE에 의해 유도되는 세포사가 억제가 되었기에 산화적 활성화가 HNE에 의해 유도된 세포사와 관련이 있음을 알 수 있었다. 이러한 결과들은 HNE가 내피세포에서 ROS와 peroxynitrite 생성을 통하여 세포사를 일으킨다는 사실을 뒷받침해 준다.

• Restorative Dentistry and Endodontics
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• 제39권2호
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• pp.89-94
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• 2014

Objectives: The aim of this study was to evaluate the cytotoxicity, setting time and compressive strength of MTA and two novel tricalcium silicate-based endodontic materials, Bioaggregate (BA) and Biodentine (BD). Materials and Methods: Cytotoxicity was evaluated by using a 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide (XTT) assay. Measurements of 9 heavy metals (arsenic, cadmium, chromium, copper, iron, lead, manganese, nickel, and zinc) were performed by inductively coupled plasma-mass spectrometry (ICP-MS) of leachates obtained by soaking the materials in distilled water. Setting time and compressive strength tests were performed following ISO requirements. Results: BA had comparable cell viability to MTA, whereas the cell viability of BD was significantly lower than that of MTA. The ICP-MS analysis revealed that BD released significantly higher amount of 5 heavy metals (arsenic, copper, iron, manganese, and zinc) than MTA and BA. The setting time of BD was significantly shorter than that of MTA and BA, and the compressive strength of BA was significantly lower than that of MTA and BD. Conclusions: BA and BD were biocompatible, and they did not show any cytotoxic effects on human periodontal ligament fibroblasts. BA showed comparable cytotoxicity to MTA but inferior physical properties. BD had somewhat higher cytotoxicity but superior physical properties than MTA.

• 대한한의학방제학회지
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• 제26권3호
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• pp.207-221
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• 2018

Objectives : Present study investigated hepatoprotective effect of Haegan-jeon extract (HE) and tried to elucidate molecular mechanism involved. According to molecular mechanism, present study optimized herbal composition of HE (op-HE) and compared in vitro and in vivo hepatoprotective effects of op-HE to HE. Methods : For in vitro experiments, HepG2 cells were exposed to arachidonic acid (AA, $10{\mu}M$) and iron ($5{\mu}M$) for inducing oxidative stress. Cell viability, GSH contents, $H_2O_2$ production, mitochondrial membrane potential, immunoblot and reporter gene assay were performed to investigate cytoprotective effects and responsible molecular mechanisms. For in vivo experiments, hepatoprotective effect of HE and op-HE were assessed on $CCl_4-induced$ liver injury mice model. Results : HE pretreatment prevented AA+iron-mediated hepatocytes apoptosis. In addition, AA+iron-induced mitochondrial dysfunction, $H_2O_2$ production, glutathione depletion were reduced by HE pretreatment. In addition, nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation, antioxidant response element (ARE)-driven reporter gene activity, and antioxidant genes expression were increased by HE. Based on reporter gene and MTT assays, we found that op-HE consisting three medicinal herbs also significantly increased transactivation of Nrf2 and reduced the AA+iron-mediated cytotoxicity. Moreover, in $CCl_4-induced$ liver injury mice model, HE-op had an ability to ameliorate $CCl_4-mediated$ increases in serum alanine transferase and aspartate aminotransferase activity, hepatic degeneration, inflammatory cell infiltration, and collagen deposition. Hepatoprotective effects of op-HE were comparable to those of HE. Conclusions : Present study suggests that op-HE as well as HE exhibit hepatoprotective effect against oxidative stress-mediated liver injury via Nrf2 activation.

• 한국식품위생안전성학회지
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• 제32권1호
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• pp.75-81
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• 2017

본 연구에서는 고등어 어육을 고감도로 보다 신속하게 검출하기 위하여 고등어 어육 중 TSSP를 이용해 단클론성 항체를 개발하고 이를 이용하여 간접 효소면역분석법(iELISA)과 western blot의 검출한계를 확인하였다. 먼저 비 열처리와 열처리를 한 고등어 어육 중 존재하는 TSSP를 확인하고, 단백질 추출에 주로 사용되는 버퍼의 종류에 따른 추출법 효율을 확인하기 위하여 전기영동과 단백질 정량을 실시하였다. 그 결과 열처리한 추출물은 비 열처리한 추출물보다 TSSP가 2배 정도 많이 추출되었으며, TSSP로 추측되는 37 kDa 부근에 형성된 밴드의 선명함과 굵기를 육안으로 비교한 결과 carbonate buffer로 추출하였을 때 밴드가 가장 두드러지게 형성되었고, 단백질 정량 결과에서도 carbonate buffer를 이용한 추출물의 단백질 농도가 가장 높은 것을 확인하였다. 이후, 생 고등어 어육을 열처리법으로 추출하여 항원을 준비하고 6주령 BALB/c mouse에 면역한 후 세포융합 및 클로닝을 통해 3A5-1번, 2번, 9번 및 16번 4종의 hybridoma cell을 확보하였다. 이렇게 개발된 항체는 수산물, 축산물, 농산물과의 교차반응성확인을 통하여 고등어 단백질에만 특이성을 가지는 것을 확인하였다. iELISA와 western blot법의 검출한계를 확인한 결과 고등어가 1% 첨가된 수준까지 검출할 수 있으며, 특히 3A5-2와 3A5-9번 항체는 1%에서도 높은 흡광도를 나타내어 민감도가 매우 높은 항체로 확인되었다. 따라서, 개발된 고등어 TSSP 특이항체를 이용한 iELISA법과 western blot법은 가공품에 혼입될 수 있는 식품 알레르겐인 고등어를 보다 신속하고 민감하게 분석할 수 있는 분석 도구로서 활용이 가능할 것으로 판단되며, 개발된 항체는 고감도 바이오센서 개발에 충분히 활용이 가능한 바이오인자로 확인되었다.

• 한국산학기술학회논문지
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• 제16권6호
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• pp.4328-4334
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• 2015

본 연구는 선박평형수처리장치 성능을 평가하기 위한 일환으로 Evan blue, Aniline blue 그리고 CMFDA 염색방법을 활용하여 해양부유생물의 생사판별을 이해하고자 하였다. Evans blue와 Aniline blue의 염색은 죽은 생물을 파란색으로 염색하여 죽은 생물을 평가하는 방법이고, CMFDA의 방법은 살아있는 생물을 녹색으로 염색하여 평가한다. 현장 동 식물플랑크톤 군집을 대상으로 Evans blue와 Aniline blue의 방법을 적용한 결과, 죽은 생물을 90%이상 염색시키는 것으로 나타났다. 하지만, 살아있는 일부 식물플랑크톤의 군집에서도 파란색으로 염색 되어, 실제 선박평형수 처리장장치의 성능을 평가하기에는 일정의 한계성을 나타내었다. 한편, 살아있는 생물을 염색하는CMFDA방법을 적용한 결과, 현장의 부유생물의 70%의 염색효과를 보였다. CMFDA방법은 FDA방법과 유사하게 살아있는 생물을 녹색으로 염색하여 생물 생사판별이 가능하게 함으로, 두 가지 방법을 중복염색(double staining)하는 방법을 고안해 보다 높은 효율의 생사판별방법을 찾고자 노력하였다. 그 결과, 두 가지의 염색시약을 중복으로 염색한 실험군에서 95%이상의 높은 효율을 보였고. 이는 단일 염색한 실험군보다 상대적으로 높은 염색효율을 얻을 수 있었다. 따라서 CMFDA+ FDA를 중복염색한 평가법이 해양생물의 생사를 판별하는데 보다 높은 효율을 보였고, 선박평형수처리장치의 성능을 평가하는 방법으로 활용이 가능할 것으로 판단된다.

• Journal of Gastric Cancer
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• 제23권3호
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• pp.476-486
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• 2023

Purpose: The optimal tumor mutational burden (TMB) value for predicting treatment response to programmed cell death-1 (PD-1) checkpoint inhibitors in advanced gastric cancer (AGC) remains unclear. We aimed to investigate the optimal TMB cutoff value that could predict the efficacy of PD-1 checkpoint inhibitors in AGC. Materials and Methods: Patients with AGC who received pembrolizumab or nivolumab between October 1, 2020, and July 27, 2021, at Samsung Medical Center in Korea were retrospectively analyzed. The TMB levels were measured using a next-generation sequencing assay. Based on receiver operating characteristic curve analysis, the TMB cutoff value was determined. Results: A total 53 patients were analyzed. The TMB cutoff value for predicting the overall response rate (ORR) to PD-1 checkpoint inhibitors was defined as 13.31 mutations per megabase (mt/Mb) with 56% sensitivity and 95% specificity. Based on this definition, 7 (13.2%) patients were TMB-high (TMB-H). The ORR differed between the TMB-low (TMB-L) and TMB-H (8.7% vs. 71.4%, P=0.001). The progression-free survival and overall survival (OS) for 53 patients were 1.93 (95% confidence interval [CI], 1.600-2.268) and 4.26 months (95% CI, 2.992-5.532). The median OS was longer in the TMB-H (20.8 months; 95% CI, 2.292-39.281) than in the TMB-L (3.31 months; 95% CI, 1.604-5.019; P=0.049). Conclusions: The TMB cutoff value for predicting treatment response in AGC patients who received PD-1 checkpoint inhibitor monotherapy as salvage treatment was 13.31 mt/Mb. When applying the programmed death ligand-1 status to TMB-H, patients who would benefit from PD-1 checkpoint inhibitors can be selected.

• 대한핵의학회지
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• 제29권3호
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• pp.332-342
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• 1995

This study was designed to evaluate the effects of various factors on the therapeutic effect of the I-131 labeled anti-carcinoembryonic antigen monoclonal antibody(anti-CEA antibody). Tetrazolium-based colorimetric assay (MTT) was used to compare in vitro cytotoxicity of 3 Korean colon cancer cell lines (SNU-C2A, SNU-C4, SNU-C5) for selection of proper 2 cell lines in this study. The changes of the size of tumor which was xenografted to nude mice (balb/c nu/nu) were compared in 4 groups (group treated I-131 labeled anti-CEA antibody, group treated with non-radiolabeled anti-CEA antibody, group treated with I-131 labeled anti-human chorionic gonadotropin monoclonal antibody (anti-hCG antibody) as nonspecific antibody, and group injected with normal saline as a control). Immunohistochemical staining and in vivo autoradiography were performed after excision of the xenografted tumor. The results were as below mentioned. The in vitro cytotoxic effect of I-131 labeled anti-CEA antibody is most prominent in SNU-C5 cell line between 3 cancer cell lines. The changes of xenografted tumor size in both SNU-C4 and SNU-5S cell tumors at the thirteenth day after injection of the antibodies were smallest in the group treated with I-131 labeled anti-CEA antibody (SNU-C4/SNU-C5; 324/342%) comparing with other groups, group treated with anti-CEA antibody (622/660%), group treated with I-131 anti-hCG antibody (538/546%), and control group(1030/724%)(P<0.02 in SNU-C4 and P<0.1 in SNU-C5 at the 13th day after injection of antibodies). On the thirteenth day after injection of the antibodies nude mice were sacreficed to count the radiouptake of tumor and to check the changes of tumor size. Correlations between radiouptake and change of tumor size were calculated in each groups and significant negative correlation was only obtained in the group treated with I-131 anti-CEA antibody (p<0.05). There were no correlations between antigenic expression of carcinoembryonic antigen and distribution of anti-CEA antibody in both SNU-C4 and SNU-C5 cell tumors on immunoperoxidase staining. On in vivo autoradiography the distributions of anti-CEA antibody were heterogeneous and the intensities of binding were various in SNU-C4 and SNU-C5 cell tumors. It is concluded that I-131 labeled tumor-specific monoclonal antibody, anti-CEA antibody is effective in suppressing the xenografted tumor growth and the effect is influenced by sensitivity of tumor cell itself to the radiolabeled antibody and other local factors instead of specificity of antibody.

• Journal of Nutrition and Health
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• 제7권3호
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• pp.1-13
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• 1974

The relative state of human iron storage may be ascertained more reliably through determination of the serum iron, iron binding capacity, transferrin saturation and absorption of radioactive iron in conjunction with studies of red cell morphology than from the study of red cell morphology alone. Recent investigations have shown that there is an increase in red cell protoporphyrin concentration in iron deficiency anemia. The significance of the red cell protoporphyrin has been discussed greatly during the years since its discovery. Two of the main factors which appear to influence the amaunt of protoporphyrin are increased erythropoiesis and factors interfering with the utilization of iron in the synthesis of hemoglobin, and iron deficiency. Recently Heller et al. have described a simplified method for blood protoporphyrin assay and this technique could be used assess nutritional iron status, wherein even minor insufficiencies are detectable as increased protoporphyrin concentrations. Based on the evaluation of the relationship between nutritional iron status and red cell protoporphyrin as an index suitable for the detection of the iron deficiency is described in this paper. RESULTS 1. Hemoglobin Concentrations and Anthropometric Measurements. The mean and standard deviations of the various anthropometric measurements of different age and sex groups are shown in table 1. There measurements have been compared with the Korean Standard. In the absence of local standards for arm circumference and skin-fold thickness over triceps, they have been compared with the standard from Jelliffe. Table 2,3, and 4 give anthropometric measurements and frequency (%) of anemia in children surveyed. The mean height of the children studid was 10 to 20 percent; below the Korean Standard. The distribution of height below 80 percent of the Standard was 21.2 percent, however, among anemic group this percentage was 27.7 percent. In general, the mean weight of the children was 10 to 15 percent below the Korean Standard. The percentage of children with weight less than 80 percent of the Standard was about 35 percent. But in the anemic group of the children, this percentage was 44 percent. The mean arm circumference was about 15 percent lower than the Jelliffe's standard. 61.2 percent of the children had values of arm circumference below 80 percent of the standard. Children with low hemoglobin levels, this percentage was 80 percent. The mean skinfold thickness over the triceps of the children studied was about 25 Percent lower than the Jelliffe's standard and 61.2 percent of the children had the value less than 80 percent of the standard. Among anemic children, this percentage was 70.8%. As may be seen from table 5, the mean hemoglobin concentration of the total group was 11.3g/100ml. Hemoglobin concentration was less than 11.0g/100ml. in 65(36.5%) of the 178 children. The degree of anemia in most of these children was mild with a hemoglobin level of less than 8.0g/100ml. found in only one child. In general, the prevalence of anemia was high in female children than male and decreased its frequency with increasing age. Relatively close relationship was observed between hemoglobin level and anthrophometric measurements especially high between arm circumference and skinfold thickness and hemoglobin but very low in height and low in weight and hemoglobin level, estimated by chi-square value. II. Serum iron, Transferrin saturation (1) Serum iron, and transferrin saturation Serum iron, transferrin saturation and red cell protoporphyrin concentrations were estimated in sub-sample of 84 children from 1 to 6 years and 24 older children between 7 and 13 years of age. The findings are presented in table 6. The mean serum iron concentration of the total group was 59ug/100ml. However, the level incrased with age from 36.6ug/100ml. (1-3years) to 80.8ug/100ml. (7-13 years). 60 percent of these children had a serum iron level less than 50ug/10ml. in the 1-3 years age group and 31.4 percent for 4-6 years group. These contrast with the finding of 12.5 percent anemic children in the 7-13 years age group. The mean transferrin saturation for the total group was 18.1 percent and frequency of anemia by transferrin saturation was observed same pattern as serum iron concentration. (2) Red cell protoporphyrin concentrations. (a) Red cell protoporphrin levels of children: Red cell protoporphyrin and other biochemical data are shown in table 4. The mean concentration in red cell of all children was fround 46.3ug/100ml. RBC. and differences with age groups were observed; in the age group 1-3 years, the mean concentration was $59.5{\pm}32.14$ ug/100ml. RBC; 4-6 years $44.1{\pm}22.57$ ug/100ml. RBC. and 7-13 years, $39.0{\pm}13.56$ ug/100ml. RBC. (b) Normal protoporphyrin values in adults: It was observed that in 10 normal adult males studied here the level of protoporphyrin in red cell ranged from 18 to 54 ug/100ml. RBC. and the mean concentration was $47.5{\sim}14.47$ ug/100ml. RBC. Other biochemical determination made on the same subjects are presented in table 8. (c) Red tell protoporphyrin concentration of occupational blood donors: The results of analyses for red cell protoporphyrin as well as serum iron, transferrin saturation and hemoglobin in the 76 blood donors are presented in table 7 and 8. In this experiment, donors were selected at random, however, most of them bled repeatedly because of poor economic situation, I doubt. Table 9 shows the distribution of red cell protoporphyrin concentration and hemoglobin concentration of occupational donors. The mean hemoglobin value for the total was 11.9 g/100 ml. When iron deficiency anemia is defined as a transferrin saturation below 15%, prevalence of anemia was 47.4 percent and the mean serum iron was 27.1ug/100ml. and red cell protoporphyrin, 168.3ug/100ml. RBC. However, mean serum iron and protoporphyrin concentration of above 15% transferrin saturation were 11.6 ug/100 ml. and 58.8 ug/100 ml. RBC. respectively. The mean Protoporphyrin concentration of non-anemic (above 15% transferrin saturation) donors was slightly higher than the results of normal adult males.

• 생명과학회지
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• 제27권6호
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• pp.673-679
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• 2017

미세소관을 따라 이동하는 모터단백질들은 세포내 물질수송에 필수적인 역할을 한다. Kinesin 1은 세포내에서 미세소관을 따라 움직이는 모터단백질로서 다양한 소포, mRNA, 그리고 단백질의 세포내 수송에 관여한다. Kinesin 1은 2개의 장쇄단위체(KHCs, 또는 KIF5s)와 2개의 경쇄단위체(KLCs)로 구성되어 있다. KIF5s는 N-말단에 모터도메인을 가지고 있고 C-말단의 운반체 결합도메인을 통해 다양한 운반체와 결합한다. 본 연구에서 KIF5B와 결합하는 단백질을 분리하기 위하여 효모 two-hybrid 탐색을 수행한 결과 미세소관의 plus end 결합단백질인 cytoplasmic linker protein 170 (CLIP-170)을 분리하였다. CLIP-170의 coiled-coil 도메인은 KIF5B의 운반체 결합도메인과 결합하였다. 또한 CLIP-170은 KIF5A와 KIF5C와도 결합하였다. 그리고 glutathione S-transferase (GST) pull-down을 통해 KIF5s와 CLIP-170이 단백질수준에서 결합함을 확인하였다. 생쥐 뇌파쇄액을 KIF5B 항체로 면역침강한 결과 CLIP-170이 같이 침강함을 확인하였다. 이러한 결과들은 kinesin 1이 세포내에서 CLIP-170을 운반함을 시사한다.

• Molecular & Cellular Toxicology
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• 제3권2호
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• pp.90-97
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• 2007

8-Hydroxyquinoline is used as antibacterial agent and antioxidant based on its function inducing the chelation of ferrous ion present in host resulting in production of chelated complex. This complex being transported to cell membrane of bacteria and fungi exerts antibacterial and antifungal action. In this study, we have carried out in vitro genetic toxicity tests and microarray analysis to understand the underlying mechanisms and the mode of action of toxicity of 8-hydroxyquinoline. TA1535 and TA98 cells were treated with 8-hydroxyquinoline to test its toxicity by basic genetic toxicity test, Ames and two new in vitro micronucleus and COMET assays were applied using CHO cells and L5178Y cells, respectively. In addition, microarray analysis of differentially expressed genes in L5178Y cells in response to 8-hydroxyquinoline were analyzed using Affymatrix genechip. The result of Ames test was that 8-hydroxyquinoline treatment increased the mutations in base substitution strain TA1535 and likewise, 8-hydroxyquinoline also increased mutations in frame shift TA98. 8-Hydroxyquinoline increased micronuclei in CHO cells and DNA damage in L5178Y. 8-Hdroxyquinoline resulted in positive response in all three tests showing its ability to induce not only mutation but also DNA damage. 783 Genes were initially selected as differentially expressed genes in response to 8-hydroxyquinoline by microarray analysis and 34 genes among them were over 4 times of log fold changed. These 34 genes could be candidate biomarkers of genetic toxic action of 8-hydroxyquinoline related to induction of mutation and/or induction of micronuclei and DNA damage. Further confirmation of these candidate markers related to their biological function will be useful to understand the detailed mode of action of 8-hydroxyquinoline.