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Radiation-induced Apoptosis, Necrosis and G2 Arrest in Fadu and Hep2 Cells

  • Lee Sam-Sun;Kang Beom-Hyun;Choi Hang-Moon;Jeon In-Seong;Heo Min-Suk;Choi Soon-Chul
    • Imaging Science in Dentistry
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    • v.30 no.4
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    • pp.275-279
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    • 2000
  • Purpose: Radiation damage is produced and viable cell number is reduced. We need to know the type of cell death by the ionizing radiation and the amount and duration of cell cycle arrest. In this study, we want to identified the main cause of the cellular damage in the oral cancer cells and normal keratinocytes with clinically useful radiation dosage. Materials and Methods: Human gingival tissue specimens obtained from healthy volunteers were used for primary culture of the normal human oral keratinocytes (NHOK). Primary NHOK were prepared from separated epithelial tissue and maintained in keratinocyte growth medium containing 0.15 mM calcium and a supplementary growth factor bullet kit. Fadu and Hep-2 cell lines were obtained from KCLB. Cells were irradiated in a /sup 137/Cs γ-irradiator at the dose of 10 Gy. The dose rate was 5.38 Gy/min. The necrotic cell death was examined with Lactate Dehydrogenase (LDH) activity in the culture medium. Every 4 day after irradiation, LDH activities were read and compared control group. Cell cycle phase distribution and preG1-incidence after radiation were analyzed by flow cytometry using Propidium Iodine staining. Cell cycle analysis were carried out with a FAC Star plus flowcytometry (FACS, Becton Dickinson, USA) and DNA histograms were processed with CELLFIT software (Becton Dickinson, USA). Results: LDH activity increased in all of the experimental cells by the times. This pattern could be seen in the non-irradiated cells, and there was no difference between the non-irradiated cells and irradiated cells. We detected an induction of apoptosis after irradiation with a single dose of 10 Gy. The maximal rate of apoptosis ranged from 4.0% to 8.0% 4 days after irradiation. In all experimental cells, we detected G2/M arrest after irradiation with a single dose of 10 Gy. Yet there were differences in the number of G2/M arrested cells. The maximal rate of the G2/M ranges from 60.0% to 80.0% 24h after irradiation. There is no significant changes on the rate of the G0/G1 phase. Conclusion: Radiation sensitivity was not related with necrosis but cell cycle arrest and apoptosis. These data suggested that more arrested cell is correlated with more apoptosis.

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Honokiol Inhibits Nitric Oxide-Induced Apoptosis in Rabbit Articular Chondrocytes via PI-3K/AKT Pathway (Honokiol에 의한 토끼의 무릎 연골세포에서 PI-3K/AKT pathway를 통하여 nitric oxide에 의해 유도되는 세포사멸의 억제)

  • Lee, Won-Kil;Kim, Song-Ja
    • Journal of Life Science
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    • v.20 no.10
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    • pp.1443-1450
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    • 2010
  • Honokiol is a small molecular weight ligand originally isolated from the Chinese medicinal herb Magnolia officinalis, a plant used in traditional Chinese and Japanese medicine [9]. In a previous study, the effects of honokiol were shown to have anti-angiogenic, anti-invasive and anti-proliferative activities in a variety of cancers [1,3,4,11,13,17,24,29,30]. We showed previously that direct production of nitric oxide (NO) by treatment of NO donor, sodium nitroprusside (SNP), led to apoptosis in rabbit articular chondrocytes [15,16]. This study confirmed that NO-induced apoptosis was suppressed by honokiol treatment in a dose-dependent manner as determined by cell phenotype, MTT assay, Western blot analysis and FACS analysis in articular chondrocytes. Treatment of honokiol inhibited SNP-induced expression of p53 as well as DNA fragmentation in articular chondrocytes, but increased expressionof pro-caspase-3. Inhibition of SNP-induced apoptosis by honokiol treatment was rescued by LY294002, the specific inhibitors of phosphoinositide 3-kinase (PI-3K) in articular chondrocytes. Our results indicate that honokiol inhibits NO-induced apoptosis via PI-3K/AKT pathway in rabbit articular chondrocytes.

Discrimination between spontaneous and posed smile: Humans versus computers (자발적 웃음과 인위적 웃음 간의 구분: 사람 대 컴퓨터)

  • Eom, Jin-Sup;Oh, Hyeong-Seock;Park, Mi-Sook;Sohn, Jin-Hun
    • Science of Emotion and Sensibility
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    • v.16 no.1
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    • pp.95-106
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    • 2013
  • The study compares accuracies between humans and computer algorithms in the discrimination of spontaneous smiles from posed smiles. For this purpose, subjects performed two tasks, one was judgment with single pictures and the other was judgment with pair comparison. At the task of judgment with single pictures, in which pictures of smiling facial expression were presented one by one, subjects were required to judge whether smiles in the pictures were spontaneous or posed. In the task for judgment with pair comparison, in which two kinds of smiles from one person were presented simultaneously, subjects were to select spontaneous smile. To calculate the discrimination algorithm accuracy, 8 kinds of facial features were used. To calculate the discriminant function, stepwise linear discriminant analysis (SLDA) was performed by using approximately 50 % of pictures, and the rest of pictures were classified by using the calculated discriminant function. In the task of single pictures, the accuracy rate of SLDA was higher than that of humans. In the analysis of accuracy on pair comparison, the accuracy rate of SLDA was also higher than that of humans. Among the 20 subjects, none of them showed the above accuracy rate of SLDA. The facial feature contributed to SLDA effectively was angle of inner eye corner, which was the degree of the openness of the eyes. According to Ekman's FACS system, this feature corresponds to AU 6. The reason why the humans had low accuracy while classifying two kinds of smiles, it appears that they didn't use the information coming from the eyes enough.

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Multivariate Analysis of Prognostic Factors in Male Breast Cancer in Serbia

  • Sipetic-Grujicic, Sandra Branko;Murtezani, Zafir Hajdar;Neskovic-Konstatinovic, Zora Borivoje;Marinkovic, Jelena Milutin;Kovcin, Vladimir Nikola;Andric, Zoran Gojko;Kostic, Sanja Vladeta;Ratkov, Isidora Stojan;Maksimovic, Jadranka Milutin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.3233-3238
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    • 2014
  • Background: The aim of this study was to analyze the demographic and clinical characteristics of male breast cancer patients in Serbia, and furthermore to determine overall survival and predictive factors for prognosis. Materials and Methods: In the period of 1996-2006 histopathological diagnosis of breast cancer was made in 84 males at the Institute for Oncology and Radiology of Serbia. For statistical analyses the Kaplan-Meier method, long-rank test and Cox proportional hazards regression model were used. Results: The mean age at diagnosis with breast cancer was $64.3{\pm}10.5$ years with a range from 35-84 years. Nearly 80% of the tumors showed ductal histology. About 44% had early tumor stages (I and II) whereas 46.4% and 9.5% of the male exhibitied stages III and IV, respectively. Only 7.1% of male patients were grade one. One-fifth of all patients had tumors measuring ${\leq}2cm$, and 14.3% larger than 5 cm. Lymph node metastasis was recorded in 40.4% patients and 47% relapse. Estrogen and progesterone receptor expression was positive in 66.7% and 58.3%, respectively. Among 14.3% of individuals tumor was HER2 positive. About two-thirds of all male patients had radical mastectomy (66.7%). Adjuvant hormonal (tamoxifene), systematic chemotherapy (CMF or FAC) and adjuvant radiotherapy were given to 59.5%, 35.7% and 29.8% patients respectively. Overall survival rates at five and ten years for male breast cancer were 55.0% and 43.9%, respectively. According to the multivariate Cox regression predictive model, a lower initial disease stage, a lower tumor grade, application of adjuvant hormone therapy and no relapse occurrence were significant independent predictors for good overall survival. Conclusions: Results of the treatment would be better if disease is discovered earlier and therefore health education and screening are an imperative in solving this problem.

Industrial-Scale Production of High-Purity Antihemophilic Factor IX from Human Plasma (사람 혈장으로부터 고순도 혈액응고 제9인자의 산업적 생산)

  • Kang, Yong;Choi, Yong-Woon;Sung, Hark-Mo;Sohn, Ki-Whan;Shin, Jeong-Sup;Kim, In-Seop
    • KSBB Journal
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    • v.23 no.1
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    • pp.37-43
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    • 2008
  • The use of antihemophilic factor IX complex has been associated with a variety of thrombotic complications, the major cause of which was the contamination of thrombogenic proteins such as vitamin K-dependent clotting factors II, VII, and X. In order to produce a commercial factor IX (GreenNine VF) free from thrombogenic potential, industrial-scale production process for high-purity factor IX from human plasma has been developed. The purification process contains cryo-precipitation, DEAE-sephadex A-50 anion-exchange chromatography, DEAE-toyopearl 650M anion-exchange column chromatography, heparin-sepharose 6FF affinity column chromatography, and CM-sepharose FF cation-exchange column chromatography. Also the process includes two viral inactivation and removal procedures, solvent/detergent treatment and nanofiltration using Viresolve NFP filter. The purification yield was 35.4%. The specific activity in the purified concentrate was 190.8 IU/mg which exceeded that in the factor IX complex (FacNine) by a factor of 48. The activities of factor II, VII, and X were not detected in GreenNine VF. SDS-PAGE analysis showed that GreenNine VF had the highest purity in comparison with commercially available high purity factor IX concentrates, Mononine, Octanyne, Berinin HS, and Immunine STIM plus 600. One batch size of the production was 2,400 vials of 250 IU product or 1,200 vials of 500 IU product from 1,600 L cryo-poor plasma.

Development of Facial Expression Recognition System based on Bayesian Network using FACS and AAM (FACS와 AAM을 이용한 Bayesian Network 기반 얼굴 표정 인식 시스템 개발)

  • Ko, Kwang-Eun;Sim, Kwee-Bo
    • Journal of the Korean Institute of Intelligent Systems
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    • v.19 no.4
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    • pp.562-567
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    • 2009
  • As a key mechanism of the human emotion interaction, Facial Expression is a powerful tools in HRI(Human Robot Interface) such as Human Computer Interface. By using a facial expression, we can bring out various reaction correspond to emotional state of user in HCI(Human Computer Interaction). Also it can infer that suitable services to supply user from service agents such as intelligent robot. In this article, We addresses the issue of expressive face modeling using an advanced active appearance model for facial emotion recognition. We consider the six universal emotional categories that are defined by Ekman. In human face, emotions are most widely represented with eyes and mouth expression. If we want to recognize the human's emotion from this facial image, we need to extract feature points such as Action Unit(AU) of Ekman. Active Appearance Model (AAM) is one of the commonly used methods for facial feature extraction and it can be applied to construct AU. Regarding the traditional AAM depends on the setting of the initial parameters of the model and this paper introduces a facial emotion recognizing method based on which is combined Advanced AAM with Bayesian Network. Firstly, we obtain the reconstructive parameters of the new gray-scale image by sample-based learning and use them to reconstruct the shape and texture of the new image and calculate the initial parameters of the AAM by the reconstructed facial model. Then reduce the distance error between the model and the target contour by adjusting the parameters of the model. Finally get the model which is matched with the facial feature outline after several iterations and use them to recognize the facial emotion by using Bayesian Network.

The Effects of Hantaan Virus on the Expression of Platelet Activating Factor Receptor and on the Activity of Platelet Activating Factor Acetylhydrolase (한탄바이러스가 혈소판활성인자 수용체 발현 및 혈소판활성인자 분해효소 활성에 미치는 영향)

  • Hwang, Ji-Young;Park, Jong-Won;Hong, Sae-Yong;Park, Ho-Sun
    • Journal of Yeungnam Medical Science
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    • v.25 no.1
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    • pp.41-49
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    • 2008
  • Background : The central physiological derangement of hemorrhagic fever with renal syndrome (HFRS) caused by hantaan virus (HTNV) is a vascular dysfunction, manifested by hemorrhage, impaired vascular tone and increased vascular permeability. Platelet activating factor (PAF), whose actions are mediated through a specific receptor, is a potent bioactive lipid. PAF has diverse biological functions in the vascular system, such as increasing vascular permeability, adhesion of leukocytes to the endothelium and reduction of cardiac output, which result in hypotension and shock. The goal of the present study was to investigate whether PAF is involved in the pathogenesis of HFRS. For this purpose, we evaluated the effect of HTNV on the expression of PAF receptor (PAF-R) and on the activity of PAF-acetylhydrolase (PAF-AH) instead of PAF because PAF is rapidly degraded by PAF-AH in vivo. Materials and methods : To evaluate the expression of PAF-R, we performed reverse-transcription PCR, western blot and FACS analyses using HTNV-infected human umbilical vein endothelial cells (HUVECs) and non-infected (control) HUVECs. In addition, we measured the activity of plasma PAF-AH in HFRS patients and normal healthy persons. Results : The mRNA and protein expression of PAF-R was increased in HTNV-infected HUVECs compared with control HUVECs at 2 and 3 days post-infection (d.p.i.). FACS analysis showed that HTNV induced the surface expression of PAF-R in HUVECs from 2 d.p.i. The activity of plasma PAF-AH was 2.5-fold lower in HFRS patients than in normal healthy persons. Conclusion : Increased PAF-R expression by HTNV might increase the responsiveness to PAF in endothelial cells. Reduced PAF-AH activity in the blood of HFRS patients might delay PAF degradation. These results suggest that changes in PAF-R and PAF-AH by HTNV might influence to PAF activity and might be involved in the vascular dysfunction of HFRS.

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Anti-cancer effect of farrerol induced apoptosis through activating p38 MAPK in Human breast cancer MCF-7 cells (인간 유방암 세포주 MCF-7에 대한 farrerol의 p38 MAPK 활성화와 세포사멸 유도를 통한 항암 효과)

  • Chae, Jongbeom;Lee, Seul Gi;Nam, Ju-Ock
    • Journal of Applied Biological Chemistry
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    • v.63 no.2
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    • pp.147-152
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    • 2020
  • Farrerol is a flavanone isolated from the traditional Chinese herb 'Man-shan-hong' (Rhododendron dauricum L.). Farrerol has been reported to have various bioactivities including anti-oxidant, anti-inflammation, and anti-fungal. However, anti-cancer effect of farrerol has not yet been reported in MCF-7 breast cancer cells. In the present study, we investigated the anti-cancer effect of farrerol on MCF-7 cells. Farrerol decreased viability and induced apoptosis of MCF-7 cells in a dose dependent manner. Ferrerol exhibited a significant anti-proliferation effect with a half-maximal inhibitory concentration (IC50) values of 145.04±1.4 μM in MTT assay, when MCF-7 cells were treated with ferrerol for 48 h. Also, ferrerol induced apoptotic bodies of MCF-7 cells as evaluated by TUNEL assay and Annexin V/PI staining using FACS. By mechanism of action, ferrerol regulated the activation of p38 mitogen-activated protein kinase and altered the expression level of BAX, Bcl-2, and Poly ADP Ribose Polymerase in MCF-7 cells. In summary, our finding demonstrated that ferrerol has anti-cancer effect through regulating the activation and expression of apoptosis-related proteins in MCF-7 cells.

The Effect of Elastic Theraband Exercise Based of PNF L/E Pattern on the Gait of the Chronic Hemiplegic Patients (고유수용성 신경근 촉진법 하지 패턴에 기초한 탄력밴드 훈련이 만성 편마비 환자의 보행에 미치는 영향)

  • Kim, Jwa-Jun;Kim, Gwang-Il;Kim, Do-Whan;Sung, Yong-In;Shin, Seung-Je
    • PNF and Movement
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    • v.5 no.2
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    • pp.47-54
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    • 2007
  • Purpose : The purpose of this study were to determine the effect of a Elastic Theraband Exercise Based of PNF L/E pattern on the gait of the chronic Hemiplegic Patients. Methods : We selected the 20 chronic Hemiplegic Patients not given treatment now and divided them into two groups of both 10 Elastic Theraband group and 10 Self Exercise. The first group went through a Elastic Theraband Exercise Based of PNF L/E pattern 30 minutes a day, 5 times a week, for 6 weeks. Exercise used to blue elastic band which 2 patterns of PNF by 1) hip extension - abduction - internal rotation with knee extension. 2) hip flexion - adduction - external rotation with knee flexion. The latter group experienced Self Exercise, 30 minutes a day, 5 times a week, for 6 weeks. Firstly, we measured the absolute improvement of gait velocity(m/s), cadence(steps/min) among walking characters. Secondly, we measured the functional walking ability such as Functional Ambulatory Category(FAC, score out of 5), Modified Motor Assesment Scale(MMAS, score out of 6). Data analysis was performed with using SPSS 12.0 win program. The descriptive analysis was used to obtain average and standard deviation. The independent t-test and the paired t-test were used to compare both the groups about pre and post training test. Treatment effects were established by pre and post assessment. Subjects tolerated the training well without side-effects. Therefore, the results of this study were as follows; Results : 1. There was a more significant improvement of Gait velocity(0.12m/s) Elastic Theraband group(p<.05). 2. There was a more significant improvement of cadence(9.40steps/min) Elastic Theraband group(p<.05). Conclusion : As we can see from above, the findings suggest that Elastic Theraband may be more effective than the Self Exercise for improving some gait parameters such as Gait velocity and Cadency. This conclusion also suggest that Elstic Theraband is more effective for the improvement of gait of chronic Hemiplegic Patients.

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Camphor Inhibits Adipocyte Differentiation via Its Impact on SMO-dependent Regulation of Hedgehog Signaling (Camphor의 Hedgehog 신호 SMO 조절을 통한 지방구세포 분화 억제효과)

  • Choi, Jae Young;Lim, Jong Seok;Lee, Ja Bok;Yang, Yung Hun
    • Journal of Life Science
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    • v.30 no.11
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    • pp.973-982
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    • 2020
  • In this study, we examined inhibition of adipocyte differentiation associated with the administration of camphor, a substance identified in extracts of the flowering plant Chrysanthemum indicum L. (CI). No camphor-mediated cytotoxicity was observed over a period of 1-10 days in studies targeting cells of the 3T3-L1 adipocyte-like line. Experiments that featured siRNA-mediated suppression of the transmembrane proteins Patched (PTCH) and Smoothened (SMO) resulted in inhibition and activation of differentiation, respectively. Interestingly, inhibition of PTCH typically activates SMO protein targeting and serves to activate hedgehog (HH)-mediated signaling. The results of our study suggest that activation of HH-mediated signaling can inhibit adipocyte differentiation. Furthermore, expression of glioma-associated oncogene homologue 1 (Gli1) was detected by flow cytometry in 62.7±1.5% of cells in response to administration of Lactobacillus rhamnosus (KCTC 3237) and in 60.4±2.2% of cells in response to camphor; these levels are higher than those detected in undifferentiated controls (24.9±3.1%). No change in the state of fermented camphor was identified by gas chromatography-mass spectrometry (GC-MS), but a 15.41% quantitative increase was confirmed in KCTC 3237. Overall, we conclude that administration of camphor resulted in overexpression of SMO and modulated the differential expression of Gli1. Animal studies focused on the impact of camphor as an agent to counteract obesity might be considered in the future. Indeed, camphor and similar physiologically active compounds from fermented CI might be developed as new and effective treatments for obesity.