• Title/Summary/Keyword: 막응축

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Morphological Study on the TUNEL Reaction of Mouse Ovary by X-ray Irradiation (방사선 조사선량에 따른 생쥐 난소의 TUNEL 염색반응에 관한 형태학적 연구)

  • Yoon, Chul-Ho;Chang, Byung-Soo
    • Applied Microscopy
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    • v.37 no.2
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    • pp.135-142
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    • 2007
  • This research investigated morphological changes of ovarian follicle according to dose of irradiation when adult mice were exposed to X-rays from 6 MeV LINAC. At day 3 after irradiation of 200 cGy, 400 cGy and 600 cGy X-ray to the hole body of mice, the ovaries collected and stained with TUNEL. The normal follicles and atretic follicles were identified to apoptosis by the staining with TUNEL. In the atretic follicles of the normal ovary, the apoptotic bodies were well appeared and stained brown color. Almost of the follicles following irradiation are stained with TUNEL, but the sensitivity of reaction is weaker than that in irradiation of 400 cGy and 600 cGy X-ray. The granulosa cells of the radiated normal follicle by 400 cGV are shown brown color. In this stage, the nucleus of granulosa cells in the atrectic follicles are condensed and picknotic feature. The size of the radiated follicle by 600 cGy are decreased than the normal follicles. The atropic follicles are filled with apoptotic bodies which change of granulosa cells and theca cells by influence of X-ray. All of cell in the follicles are strongly positive stained with TUNEL by irradiation of 600 cGy.

Structural and Functional Roles of Caspase-8 in Extrinsic Apoptosis (Apoptosis의 외인성 경로에서 caspase-8의 구조적 및 기능적 역할)

  • Ha, Min Seon;Jeong, Mi Suk;Jang, Se Bok
    • Journal of Life Science
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    • v.31 no.10
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    • pp.954-959
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    • 2021
  • Apoptosis is an important mechanism that regulates cellular populations to maintain homeostasis, and the caspases, a family of cysteine proteases, are key mediators of the apoptosis pathway. Caspase-8 is an initiator caspase of the extrinsic apoptotic pathway, which is initiated by extracellular stimuli. Caspase-8 have two conserved domains, N-terminal tandem death effector domains (DED) and C-terminal two catalytic domain, which are important for this extrinsic apoptosis pathway. In extrinsic apoptosis pathway, death receptors which members of TNF superfamily are activated by binding of death receptor specific ligands from cell outside. After the activated death receptors recruit adaptor protein Fas-associated death domain protein (FADD), death domains (DD) of death receptor and FADD bind to each other and FADD combined with death receptor recruits procaspase-8, a precursor form of caspase-8. The DED of FADD and procaspase-8 bind to one another and FADD-bound procaspase-8 is activated by cleavage of the prodomain. This death receptor-FADD-caspase-8 complex called death inducing signaling complex (DISC). Cellular FLICE-inhibitory proteins (c-FLIPs) regulate caspase-8 activation by acting both anti- and pro-apoptotically, and caspase-8 activation initiates the activation of executioner caspases such as caspase-3. Finally activated executioner caspases complete the apoptosis by acting critically DNA degradation, nuclear condensation, plasma membrane blebbing, and the proteolysis of certain caspase substrates.

Signals of MLCK and ROCK Pathways Triggered via Lymphotoxin β Receptor are Involved in Stress Fiber Change of Fibroblastic Reticular Cells (FRC에서 Lymphotoxin β receptor의 자극은 MLCK와 ROCK의 이중 신호전달 경로를 통해 stress fiber 변화에 관여)

  • Kim, Dae Sik;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.29 no.2
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    • pp.256-264
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    • 2019
  • Lymphotoxin ${\beta}$ receptor ($LT{\beta}R$), a member of the tumor necrosis factor receptor family, plays an important role in lymphoid tissue's architecture and organogenesis. In contrast, MLCK and ROCK play critical roles in the regulation of stress fiber (SF) formation in cells. To determine whether $LT{\beta}R$ stimulation in fibroblastic reticular cells (FRCs) is involved in these signaling pathways, myosin light chain kinase inhibitor-7 (ML-7) was used to inhibit them. ML7-treated FRCs completely blocked SFs and showed retraction and shrinkage processes comparable to those observed in agonistic anti-$LT{\beta}R$ antibody-treated cells. The inhibition of ROCK activity with Y27632-induced changes in actin cytoskeleton organization and cell morphology in FRCs. Actin bundles rearranged into SFs, and phospho-myosin light chain (p-MLC) co-localized in FRCs. We checked the level of Rho-guanosine diphosphate (RhoGDP)/guanosine triphosphate (GTP) exchange activity using FRC lysate. When $LT{\beta}R$ was stimulated with agonistic anti-$LT{\beta}R$ antibodies, Rho-GDP/GTP exchange activity was markedly reduced. Regarding $LT{\beta}R$ signaling with a focus on MLCK inhibition, we showed that the phosphorylation of MLCs was reduced by $LT{\beta}R$ stimulation in FRCs. Cytoskeleton components, such as tubulin, b-actin, and phospho-ezrin proteins acting as membrane-cytoskeleton linkers, were produced in de-phosphorylation, and they reduced expression in agonistic anti-$LT{\beta}R$ antibody-treated FRCs. Collectively, the results suggested that MLCK and ROCK were simultaneously responsible for SF regulation triggered by $LT{\beta}R$ signaling in FRCs.

A study on frost prediction model using machine learning (머신러닝을 사용한 서리 예측 연구)

  • Kim, Hyojeoung;Kim, Sahm
    • The Korean Journal of Applied Statistics
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    • v.35 no.4
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    • pp.543-552
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    • 2022
  • When frost occurs, crops are directly damaged. When crops come into contact with low temperatures, tissues freeze, which hardens and destroys the cell membranes or chloroplasts, or dry cells to death. In July 2020, a sudden sub-zero weather and frost hit the Minas Gerais state of Brazil, the world's largest coffee producer, damaging about 30% of local coffee trees. As a result, coffee prices have risen significantly due to the damage, and farmers with severe damage can produce coffee only after three years for crops to recover, which is expected to cause long-term damage. In this paper, we tried to predict frost using frost generation data and weather observation data provided by the Korea Meteorological Administration to prevent severe frost. A model was constructed by reflecting weather factors such as wind speed, temperature, humidity, precipitation, and cloudiness. Using XGB(eXtreme Gradient Boosting), SVM(Support Vector Machine), Random Forest, and MLP(Multi Layer perceptron) models, various hyper parameters were applied as training data to select the best model for each model. Finally, the results were evaluated as accuracy(acc) and CSI(Critical Success Index) in test data. XGB was the best model compared to other models with 90.4% ac and 64.4% CSI, followed by SVM with 89.7% ac and 61.2% CSI. Random Forest and MLP showed similar performance with about 89% ac and about 60% CSI.

Anti-cancer activity of the ethylacetate fraction from Orostachys japonicus in A549 human lung cancer cells by induction of apoptosis and cell cycle arrest (인체 폐암 세포에 대한 와송 유래 에틸아세테이트 분획 생리 활성 물질의 세포사멸 유도 및 세포주기 억제 항암활성)

  • Kwon, Ji-Hye;Lee, Dong-Seok;Jung, Eun-Cheol;Kim, Hyeon-Mi;Kim, Su-Bin;Ryu, Deok-Seon
    • Asia-pacific Journal of Multimedia Services Convergent with Art, Humanities, and Sociology
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    • v.7 no.1
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    • pp.395-405
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    • 2017
  • To confirm potential anti-cancer activities of ethylacetate (EtOAc) fraction from Orostachys japonicus on the A549 human lung cancer cells, this study examined. As a result of conducting MTS assay for measuring cell viability, the EtOAc fraction inhibited the proliferation of A549 cells in a dose-dependent manner. To investigate whether the inhibiting A549 cell viability was caused by apoptosis, this study analyzed chromatin condensation in A549 cells using DAPI staining. The morphological changes such as the formation of nuclear condensation were formed in a dose-dependent manner. Also, this study performed Annexin V-FITC staining for detecting phosphatidylinositol (PS). As a result of Annexin V-FITC staining to investigate level of early and late apoptosis, the apoptosis level treated with EtOAc fraction was higher than that of control. RT-PCR was performed to study the correlation between G2/M cell cycle arrest and cell cycle control genes. The anti-cancer activity of EtOAc fraction was accompanied by inhibition of CDK1, 4, cyclin B1 and D1 mRNA. This study also examined the expression of various marker proteins: p53, Bax, Bcl-2 and pro-caspase 3. Western blotting revealed that p53 and Bax proteins were up-regulated, and Bcl-2 and pro-caspase 3 proteins down-regulated in a time and dose-dependent manner.

Apoptosis-Induced Effects of Extract from Artemisia annua Linné by Modulating Akt/mTOR/GSK-3β Signal Pathway in AGS Human Gastric Carcinoma Cells (AGS 인체 위암 세포에서 Akt/mTOR/GSK-3β 신호경로 조절을 통한 개똥쑥 추출물의 Apoptosis 유도 효과)

  • Kim, Eun Ji;Kim, Guen Tae;Kim, Bo Min;Lim, Eun Gyeong;Kim, Sang-Yong;Kim, Young Min
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.9
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    • pp.1257-1264
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    • 2016
  • Extracts from Artemisia annua $Linn\acute{e}$ (AAE) have various functions (anti-malaria, anti-virus, and anti-oxidant). However, the mechanism of the effects of AAE is not well known. Thus, we determined the apoptotic effects of AAE in AGS human gastric carcinoma cells. In this study, we suggested that AAE may exert cancer cell apoptosis through the Akt/mammalian target of rapamycin (mTOR)/glycogen synthase kinase (GSK)-$3{\beta}$ signal pathway and mitochondria-mediated apoptotic proteins. Activation by Akt phosphorylation resulted in cell proliferation through phosphorylation of tuberous sclerosis complex 2 (TSC2), mTOR, and GSK-$3{\beta}$. Thus, de-phosphorylation of Akt inhibited cell proliferation and induced apoptosis through inhibition of Akt, mTOR, phosphorylation of GSK-$3{\beta}$ at serine9, and control of Bcl-2 family members. Inhibition of GSK-$3{\beta}$ attenuated loss of mitochondrial membrane potential and release of cytochrome C. Bax and pro-apoptotic proteins were activated by their translocation into mitochondria from the cytosol. Translocation of Bax induced outer membrane transmission and generated apoptosis through cytochrome C release and caspase activity. We also measured 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, lactate dehydrogenase assay, Hoechst 33342 staining, Annexin V-PI staining, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide staining, and Western blotting. Accordingly, our study showed that AAE treatment to AGS cells resulted in inhibition of Akt, TSC2, GSK-$3{\beta}$-phosphorylated, Bim, Bcl-2, and pro-caspase 3 as well as activation of Bax and Bak expression. These results indicate that AAE induced apoptosis via a mitochondrial event through regulation of the Akt/mTOR/GSK-$3{\beta}$ signaling pathways.

Lymphotoxin β Receptor Stimulation Is Linked to MLCK Activity and Suppresses Stress Fiber Formation in Agonistic Anti-LTβR Antibody-stimulated Fibroblastic Reticular Cells (FRC에서 agonistic anti-LTβR antibody의 LTβR 자극은 MLCK 연관성 및 stress fiber 형성에 대한 강력한 억제 작용)

  • Kim, Min Hwan;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1199-1206
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    • 2017
  • The lymphotoxin ${\beta}$ receptor ($LT{\beta}R$), a member of the tumor necrosis factor receptor family, plays an important role in lymphoid tissue's architecture and organogenesis. We found that $LT{\beta}R$ stimulation induced changes in stress fibers (SFs) in fibroblastic reticular cells (FRCs). MLCK and ROCK play critical roles in the regulation of SF formation in cells. The present study was performed to investigate the antifibrotic effects on SF regulation of $LT{\beta}R$ signaling, with a focus on MLCK inhibition. The effect of $LT{\beta}R$ on the SF change was analyzed using immunoblot and fluorescence assays and agonistic $anti-LT{\beta}R$ antibody-treated FRCs. In addition, we checked the level of Rho-guanosine diphosphate (GDP)/guanosine triphosphate (GTP) exchange activity with FRC lysate. Phospho-ezrin proteins acting as membrane-cytoskeleton linkers completely de-phosphorylated in agonistic $anti-LT{\beta}R$ antibody-treated FRCs. The actin bundles rearranged into SFs, where phospho-myosin light chain (p-MLC) co-localized in FRCs. ML7-treated FRCs completely blocked SFs and showed retraction and shrinkage processes comparable to those observed in agonistic $anti-LT{\beta}R$ antibody-treated cells. Inhibition of ROCK activity induced changes in the actin cytoskeleton organization; however, some SFs remained in the cells, while they were completely disrupted by MLCK inhibition with ML7. We showed that the phosphorylation of MLC was completely abolished with $LT{\beta}R$ stimulation in FRCs. When $LT{\beta}R$ was stimulated with the agonistic $anti-LT{\beta}R$ antibody, the Rho-GDP/GTP exchange activity was reduced, however, the activity was not completely abolished. Collectively, the results illustrated that MLCK was potently responsible for the SF regulation triggered via $LT{\beta}R$ signaling in FRCs.

Effect of Air Circulation Velocity on the Rate of Lumber Drying in a Small Compartment Wood Drying Kiln (소형 목재인공건조실에 있어서 공기순환속도가 목재건조율에 미치는 영향)

  • Chung, Byung-Jae
    • Journal of the Korean Wood Science and Technology
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    • v.2 no.2
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    • pp.5-7
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    • 1974
  • 1. This study indicates that above the fiber saturation point the drying rate can be increased with increasing the velocity of the air circutation, i.e., the drying rate of sample boards is proportional to the air velocity, but below the fiber saturation point, the effect of the velocity of air circulation is very low as shown in Figs. 1 and 2. 2. Under the controlled temperature and humidity in the kiln, the more the sample boards have moisture, the higher drying rate of it can be obtained. In other words, this means that even though in the case of drying various moisture content of wood, at the final drying stage, approximately the same percentage of moisture content of wood can be secured by employing the higher velocity of air circulation. 3. This study shows that the rate of drying in kiln changes distinctly at the fiber saturation point, i, e., above the fiber saturation point, the drying curve shows concave aginst the X axsis, but below the fiber saturation point, in the range from 30 percent of moisture content to 20 percent of moisture content, the curve shows convex as shown in Fig. 3. As the drying progresses, however, the drying curve shows concave again below 20 percent of moisture content. This means that inflection point of drying curve may be located clearly at the fiber saturation point, i.e., 30 percent of moisture content. As mentioned above, the 30 percent of moisture content of wood at which the inflectional point appears can be recognized as a critical point, i. e., the fiber saturation point at which all free water was removed from wood. The existence of inflectional point indicates that the evaporation of hygroscopic water in a cell wall is more difficult than the evaporation of free water in a cell cavity and the minor space of cell wall. The convex curve in the range of moisture content from 30 percent to 20 percent means that the evaporation of capillary condensed water has a tendency of the same rates of drying approximately, but as approaching to the 20 percent of moisture, the transfusion of moisture from wood becomes difficult because of having less moisture in cell wall. Below 20 percent of moisture content, the drying curve shows concave again, which means that it is difficult to remove the moisture located nearer to the surface of cellulose molecules and the surface bound water. These relations were revealed in Fig. 4. In comparison AC curve which does not have the two inflection points with BD curve which has two inflection points, i.e., Band D, they are mentioned already, by existence of the inflection points, the curve BD shows that the change of drying rate in the interval from 20 percent of moisture content to 30 percent of moisture content is not greater than in the case of the curve AC in the same interval. At the inflection point of 30 percent of moisture content, it can be noticed that the changing of the drying rate is very conspicuous. This phenomenon also can be recognized, as it is noticed by the Fig. 3, the drying rate from green to 30 percent of moisture content is very great. But the inclination of the curve is very slow from 30 percent of moisture content to 20 percent of moisture content, i.e., the inclination of the curve becomes almost horizontal lines. Acknowledgments Gratitude is expressed to Fred E. Dickinson, Professor of 'Wood Technology, School of Natural Resources, University of Michigan, USA for his suggestion to carry out this study.

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The Differentiation of the Cholinergic Nerve Cells at the Meynert Basal Nucleus of the Basal Forebrains in the Growth Period of Rat (흰쥐 전뇌 기저부 Meynert기저핵에서 출생 후 발육기간에 따른 콜린성 신경세포의 분화)

  • Hahm, Young-Ok;Kim, Soo-Jin
    • Applied Microscopy
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    • v.31 no.4
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    • pp.355-365
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    • 2001
  • This study was performed to investigate the distribution and differentiation on the immunoreacted cells of the ChAT (choline acetyltransferase) at the Meynert basal nucleus of the forebrains in the growth periods of rat, using the immunohistochemistric method. According to the cell shape and the ratio of long axis vs short axis of cell soma, the ChAT antibody reacted nerve cells in the Meynert basal nucleus of the rats were classified into six types. In the adult rats, the FD (frequency distributions) of round, oval and elongated cells were maximum. The FD of these types were shown to be progressively decreased during the postnatal development. In addition, the FD of elongated nerve cells in were observed in the adult rats respectively. This was thought to be the same phenomenon as those in the round and oval cells . The total mean volume of ChAT antibody reacted cell somata was lowest in the PND (postnatal days) 7 rats and was highest in the PND 21 rats. But, those were decreased to the adult. These results suggest that ChAT antibody reacted nerve cells grow up to PND 21 and then, differentiate into the various types by neurites outgrowing. On the electron micrography, the adult rat forebrain cells were obtained to be well developed ribosomes, polysomes , rough endoplasmic reticula and mitochondria. The immunreactivities were observed in ribosomes, polysomes, rough endoplasmic reticula and outer membrane of mitochondria. Golgi complexes were poorly developed and not showed jmmunreactivity. The ribosomes , polysomes and endoplasmic reticula are considered to be closely related to the inter cellular localization and biosynthesis of the ChAT but not Golgi complex. According to the results in the present study, it is considered that the ChAT-immunoreacted nerve cells in the Meynert basal nucleus of the rat forebrains are differentiated throughout the postnatal development with following processes of changes; 1) the cholinergic nerve cells develop postnatally 2) cell soma volumes gradually increase during the early postnatal days 3) and then, cells differentiate into the various types by projecting the neurites to the appropriate area after PND 21.

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Effects of Deer Antler on the Regeneration of Peripheral Nerves; About Sprout Formation of Experimentally Transected Sciatic Nerves in Rat (말초신경의 재생에 대한 녹용의 효과; 랫드에서 실험적 절단 좌골신경의 Sprout 형성에 관해)

  • Chang, Byung-Joon;Cho, Ik-Hyun;Choi, Hye-Young;Won, Hui-Young;Park, Chang-Hyun;Bae, Chun-Sik;Choe, Nong-Hoon
    • Applied Microscopy
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    • v.32 no.1
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    • pp.67-80
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    • 2002
  • This study was carried out to investigate the effects of deer antler extract on the regeneration of peripheral nerves. Sprague-Dawley male rats weighing about 300 gm were fed deer antler extract for 1, 2, and 3 weeks per oral (1.5 ml/100 gm B.W.), respectively, once a day and transected both sides of sciatic nerve of each leg. After keeping for 6 hours, sciatic nerves taken from proximal part of transected region were treated with conventional transmission electron microscopical method and then observed with electron microscope. The results obtained were summarized as follows; 1. Sciatic nerves of normal control group were not showing any sprouts and electron dense axolemmal projections were frequently observed. 2. Sciatic nerves of saline treated groups were showing axonal sprouts at the nodes of Ranvier. The length of them was usually short, and numerous vesicles, vacuoles and organelles including neurofilament were contained. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 29 (29%) in 1 week treated group, 32 (32%) in 2 weeks treated group, and 30 (30%) in 3 weeks treated group, respectively. 3. Sciatic nerves of deer antler treated groups were showing axonal sprouts at the node of Ranvier as well. Although most of the sprouts were short, some sprouts of 2 weeks and 3 weeks treated groups were quite long. Sprouts usually contained numerous vesicles, vacuoles and cell organelles such as neurofilaments and mitochondria. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 38 (38%) in 1 week treated group, 46 (46%) in 2 weeks treated group, and 48 (48%) in 3 weeks treated group respectively. The results described above explain pretreatment of deer antler extract improves the sprout formation of transected sciatic nerves, and then it suggests deer antler may be effective for the regeneration of peripheral nerves.