• Title/Summary/Keyword: cell model

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Measuring Intracellular Mycobacterial Killing Using a Human Whole Blood Assay (인체 전혈 모델을 이용한 세포내 결핵균 살균력에 관한 연구)

  • Cheon, Seon-Hee;Song, Ho-Yeon;Lee, Eun-Hee;Oh, Hee-Jung;Kang, In-Sook;Cho, Ji-Yoon;Hong, Young-Sun
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.5
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    • pp.497-509
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    • 2002
  • Background : The mechanisms through which cellular activation results in intracellular mycobacterial killing is only partially understood. However, in vitro studies of human immunity to Mycobacterium tuberculosis have been largely modeled on the work reported by Crowle, which is complicated by several factors. The whole blood culture is simple and allows the simultaneous analysis of the relationship between bacterial killing and the effect of effector cells and humoral factors. In this study, we attempted to determine the extent to which M. tuberculosis is killed in a human whole blood culture and to explore the role of the host and microbial factor in this process. Methods : The PPD positive subject were compared to the umbilical cord blood and patients with tuberculosis, diabetes and lung cancer. The culture is performed using heparinized whole blood diluted with a culture medium and infected with a low number of M. avium or M. tuberculosis $H_{37}Ra$ for 4 days by rotating the culture in a $37^{\circ}C$, 5% $CO_2$ incubator. In some experiments, methlprednisolone- or pentoxifyline were used to inhibit the immune response. To assess the role of the T-cell subsets, CD4+, CD8+ T-cells or both were removed from the blood using magnetic beads. The ${\Delta}$ log killing ratio was defined using a CFU assay as the difference in the log number of viable organisms in the completed culture compared to the inoculum. Results : 1. A trend was noted toward the improved killing of mycobacteria in PPD+ subjects comparing to the umbilical cord blood but there was no specific difference in the patients with tuberculosis, diabetes and lung cancer. 2. Methylprednisolone and pentoxifyline adversely affected the killing in the PPD+ subjects umbilical cord blood and patients with tuberculosis. 3. The deletion of CD4+ or CD8+ T-lymphocytes adversely affected the killing of M. avium and M. tuberculosis $H_{37}Ra$ by PPD+ subjects. Deletion of both cell types had an additive effect, particularly in M. tuberculosis $H_{37}Ra$. 4. A significantly improved mycobacterial killing was noted after chemotherapy in patients with tuberculosis and the ${\Delta}$ logKR continuously decreased in a 3 and 4 days of whole blood culture. Conclusion : The in vitro bactericidal assay by human whole blood culture model was settled using a CFU assay. However, the host immunity to M. tuberculosis was not apparent in the human whole blood culture bactericidal assay, and patients with tuberculosis showed markedly improved bacterial killing after anti-tuberculous chemotherapy compared to before. The simplicity of a whole blood culture facilitates its inclusion in a clinical trial and it may have a potential role as a surrogate marker in a TB vaccine trial.

Development of Korean Version of Heparin-Coated Shunt (헤파린 표면처리된 국산화 혈관우회도관의 개발)

  • Sun, Kyung;Park, Ki-Dong;Baik, Kwang-Je;Lee, Hye-Won;Choi, Jong-Won;Kim, Seung-Chol;Kim, Taik-Jin;Lee, Seung-Yeol;Kim, Kwang-Taek;Kim, Hyoung-Mook;Lee, In-Sung
    • Journal of Chest Surgery
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    • v.32 no.2
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    • pp.97-107
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    • 1999
  • Background: This study was designed to develop a Korean version of the heparin-coated vascular bypass shunt by using a physical dispersing technique. The safety and effectiveness of the thrombo-resistant shunt were tested in experimental animals. Material and Method: A bypass shunt model was constructed on the descending thoracic aorta of 21 adult mongrel dogs(17.5-25 kg). The animals were divided into groups of no-treatment(CONTROL group; n=3), no-treatment with systemic heparinization(HEPARIN group; n=6), Gott heparin shunt (GOTT group; n=6), or Korean heparin shunt(KIST group; n=6). Parameters observed were complete blood cell counts, coagulation profiles, kidney and liver function(BUN/Cr and AST/ ALT), and surface scanning electron microscope(SSEM) findings. Blood was sampled from the aortic blood distal to the shunt and was compared before the bypass and at 2 hours after the bypass. Result: There were no differences between the groups before the bypass. At bypass 2 hours, platelet level increased in the HEPARIN and GOTT groups(p<0.05), but there were no differences between the groups. Changes in other blood cell counts were insignificant between the groups. Activated clotting time, activated partial thromboplastin time, and thrombin time were prolonged in the HEPARIN group(p<0.05) and differences between the groups were significant(p<0.005). Prothrombin time increased in the GOTT group(p<0.05) without having any differences between the groups. Changes in fibrinogen level were insignificant between the groups. Antithrombin III levels were increased in the HEPARIN and KIST groups(p<0.05), and the inter-group differences were also significant(p<0.05). Protein C level decreased in the HEPARIN group(p<0.05) without having any differences between the groups. BUN levels increased in all groups, especially in the HEPARIN and KIST groups(p<0.05), but there were no differences between the groups. Changes of Cr, AST, and ALT levels were insignificant between the groups. SSEM findings revealed severe aggregation of platelets and other cellular elements in the CONTROL group, and the HEPARIN group showed more adherence of the cellular elements than the GOTT or KIST group. Conclusion: Above results show that the heparin-coated bypass shunts(either GOTT or KIST) can suppress thrombus formation on the surface without inducing bleeding tendencies, while systemic heparinization(HEPARIN) may not be able to block activation of the coagulation system on the surface in contact with foreign materials but increases the bleeding tendencies. We also conclude that the thrombo-resistant effects of the Korean version of heparin shunt(KIST) are similar to those of the commercialized heparin shunt(GOTT).

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Effect of Boswellia serrata Extracts on Degenerative Osteoarthritis in vitro and in vivo Models (보스웰리아 추출물의 골관절염 억제 효과 연구)

  • Nam, Da-Eun;Kim, Ok Kyung;Shim, Tae Jin;Kim, Ji Hoon;Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.5
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    • pp.631-640
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    • 2014
  • The inhibitory effects of Boswellia serrata (BW) extracts on degenerative osteoarthritis were investigated in primary-cultured rat cartilage cells and a monosodium-iodoacetate (MIA)-induced osteoarthritis rat model. To identify the protective effects of BW extract against $H_2O_2$ ($800{\mu}M$, 2 hr) in vitro, cell survival was measured by MTT assay. Cell survival after $H_2O_2$ treatment was elevated by BW extract at a concentration of $20{\mu}g/mL$. In addition, BW extract treatment significantly reduced and normalized the productions of pro-inflammatory factors, nuclear transcription factor ${\kappa}B$, cyclooxygenase-2, tumor necrosis factor-${\alpha}$, and interleukin-6 at a concentration of $20{\mu}g/mL$. Treatment of chondrocytes with BW extract significantly reduced 5-lipoxygenase activity and production of prostaglandin E2, especially at a concentration of $10{\sim}20{\mu}g/mL$. For the in vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats. Consumption of a diet containing BW extract (100 and 200 mg/kg) for 35 days significantly inhibited the development and severity of osteoarthritis in rats. To determine the genetic expression of arthritic factors in articular cartilage, real-time PCR was applied to measure matrix metalloproteinases (MMP-3, MMP-9, and MMP-13), collagen type I, collagen type II, and aggrecan, and BW extract had protective effects at a concentration of 200 mg/kg. In conclusion, BW extract was able to inhibit articular cartilage degeneration by preventing extracellular matrix degradation and chondrocyte injury. One can consider that BW extract may be a potential therapeutic treatment for degenerative osteoarthritis.

Edge to Edge Model and Delay Performance Evaluation for Autonomous Driving (자율 주행을 위한 Edge to Edge 모델 및 지연 성능 평가)

  • Cho, Moon Ki;Bae, Kyoung Yul
    • Journal of Intelligence and Information Systems
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    • v.27 no.1
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    • pp.191-207
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    • 2021
  • Up to this day, mobile communications have evolved rapidly over the decades, mainly focusing on speed-up to meet the growing data demands of 2G to 5G. And with the start of the 5G era, efforts are being made to provide such various services to customers, as IoT, V2X, robots, artificial intelligence, augmented virtual reality, and smart cities, which are expected to change the environment of our lives and industries as a whole. In a bid to provide those services, on top of high speed data, reduced latency and reliability are critical for real-time services. Thus, 5G has paved the way for service delivery through maximum speed of 20Gbps, a delay of 1ms, and a connecting device of 106/㎢ In particular, in intelligent traffic control systems and services using various vehicle-based Vehicle to X (V2X), such as traffic control, in addition to high-speed data speed, reduction of delay and reliability for real-time services are very important. 5G communication uses high frequencies of 3.5Ghz and 28Ghz. These high-frequency waves can go with high-speed thanks to their straightness while their short wavelength and small diffraction angle limit their reach to distance and prevent them from penetrating walls, causing restrictions on their use indoors. Therefore, under existing networks it's difficult to overcome these constraints. The underlying centralized SDN also has a limited capability in offering delay-sensitive services because communication with many nodes creates overload in its processing. Basically, SDN, which means a structure that separates signals from the control plane from packets in the data plane, requires control of the delay-related tree structure available in the event of an emergency during autonomous driving. In these scenarios, the network architecture that handles in-vehicle information is a major variable of delay. Since SDNs in general centralized structures are difficult to meet the desired delay level, studies on the optimal size of SDNs for information processing should be conducted. Thus, SDNs need to be separated on a certain scale and construct a new type of network, which can efficiently respond to dynamically changing traffic and provide high-quality, flexible services. Moreover, the structure of these networks is closely related to ultra-low latency, high confidence, and hyper-connectivity and should be based on a new form of split SDN rather than an existing centralized SDN structure, even in the case of the worst condition. And in these SDN structural networks, where automobiles pass through small 5G cells very quickly, the information change cycle, round trip delay (RTD), and the data processing time of SDN are highly correlated with the delay. Of these, RDT is not a significant factor because it has sufficient speed and less than 1 ms of delay, but the information change cycle and data processing time of SDN are factors that greatly affect the delay. Especially, in an emergency of self-driving environment linked to an ITS(Intelligent Traffic System) that requires low latency and high reliability, information should be transmitted and processed very quickly. That is a case in point where delay plays a very sensitive role. In this paper, we study the SDN architecture in emergencies during autonomous driving and conduct analysis through simulation of the correlation with the cell layer in which the vehicle should request relevant information according to the information flow. For simulation: As the Data Rate of 5G is high enough, we can assume the information for neighbor vehicle support to the car without errors. Furthermore, we assumed 5G small cells within 50 ~ 250 m in cell radius, and the maximum speed of the vehicle was considered as a 30km ~ 200 km/hour in order to examine the network architecture to minimize the delay.

Anti-inflammatory Effects of Pentoxifylline and Neutrophil Elastase Inhibitor on Lipopolysaccharide-Induced Acute Lung Injury In Vitro (In Vitro 내독소 유도성 급성 폐손상에서 Pentoxifylline과 Neutrophil Elastase Inhibitor의 항염효과)

  • Kim, Young-Kyoon;Kim, Seung-Joon;Park, Yong-Keun;Kim, Seok-Chan;Kim, Kwan-Hyoung;Moon, Hwa-Sik;Song, Jeong-Sup;Park, Sung-Hak;Kim, Sang-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.6
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    • pp.691-702
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    • 2000
  • Background : Acute lung injury (ALI) is a commonly encountered respiratory disease and its prognosis is poor when the treatment is not provided promptly and properly. However no specific pharmacologic treatment is currently available for ALI, although recently several supportive drugs have been under scrutiny. We studied anti-inflammatory effects of pentoxifylline (PF), a methylated xanthine, and ONO-5046, a synthetic neutrophil elastase inhibitor on lipopolysaccharide (LPS)-induced ALI in vitro. Methods : To establish an in vitro model of LPS-induced ALI, primary rat alveolar macrophages and peripheral neutrophils in various ratios (1:0, 5:1, 1:1, 1:5, 0:1) were co-cultured with transformed rat alveolar epithelial cells (L2 cell line) or vascular endothelial cells (IP2-E4 cell line) under LPS stimulation. Each experiment was divided into five groups-control, LPS, LPS+PF, LPS+ONO, and LPS+PF+ONO. We compared LPS-induced superoxide anion productions from primary rat alveolar macrophages and peripheral neutrophils in various ratios, and the resultant cytotoxicity on L2 cells or IP2-E4 cells between groups. In addition we also compared the productions of tumor necrosis factor (TNF)-$\alpha$ interleukin (IL)-$1{\beta}$, monocyte chemotactic protein(MCP)-1, IL-6, and IL-10 as well as mRNA expressions of TNF-$\alpha$ inducible nitric oxide synthetase(iNOS), and MCP-1 from LPS-stimulated primary rat alveolar macrophages between groups. Results : (1) PF and ONO-5046 in each or both showed a trend to suppress LPS-induced superoxide anion productions from primary rat alveolar macrophages and peripheral neutrophils regardless of their ratio, except for the LPS+PF+ONO group with the 1:5 ratio, although statistical significance was limited to a few selected experimental conditions. (2) PF and ONO-5046 in each or both showed a trend to prevent IP2-E4 cells from LPS-induced cytotoxicity by primary rat alveolar macrophages and peripheral neutrophils regardless their ratio, although statistical significance was limited to a few selected experimental conditions. the effects of PF and/or ONO-5046 on LPS-induced L2 cell cytotoxicity varied according to experimental conditions. (3) PF showed a trend to inhibit LPS-induced productions of INF-$\alpha$ MCP-1, and IL-10 from primary rat alveolar macrophages. ONO-5046 alone didnot affect the LPS-induced productions of proinflammatory cytokines from primary rat alveolar macrophages but the combination of PF and ONO-5046 showed a trend to suppress LPS-induced productions of INF-$\alpha$ and IL-10 PF and ONO-5046 in each or both showed a trend to increase LPS-induced IL-$\beta$ and IL-6 productions from primary rat alveolar macrophages. (4) PF and ONO-5046 in each or both showed a trend to attenuate LPS-induced mRNA expressions of TNF-$\alpha$ and MCP-1 from primary rat alveolar macrophages but at the same time showed a trend increase iNOS mRNA expression. Conclusion : These results suggest that PF and ONO-5046 may play a role in attenuating inflammation in LPS-induced ALI and that further study is needed to use these drugs as a new supportive therapeutic strategy for ALI.

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Exposure Assessments of Environmental Contaminants in Ansim Briquette Fuel Complex, Daegu(I) - Effect zone of environmental pneumoconiosis and fugitive dust - (대구 안심연료단지 환경오염물질 노출 평가(I) - 환경성 진폐증 및 비산먼지 영향권역 -)

  • Jung, Jong-Hyeon;Oh, In-Bo;Phee, Young-Gyu;Nam, Mi-Ran;Hwang, Mi-Kyoung;Bang, Jin-Hee;Jeon, Soo-Bin;Lee, Sang-sup;Yu, Seung-do;KimS, Byung-Seok;Yoo, Seok-Ju;Lee, Kwan;Lim, Hyun-Sul
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.25 no.3
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    • pp.366-379
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    • 2015
  • Objectives: The objective of this study is to assess airborne particulate matter(PM) pollution and its effect on health of residents living near Ansim Briquette Fuel Complex in Daegu metropolitan region. Methods: The California Puff(CALPUFF) dispersion model, version 5.8, which can estimate the dispersion direction and range of airborn $PM_{10}$ was used to determine the possible areas affected by $PM_{10}$ pollutants emitted from Ansim briquette fuel complex. The CALPUFF modeling with 200 m grid-cell resolution was performed based on $PM_{10}$ emissions estimated from the amount of coal consumption in the fuel complex for four months in 2012. The Weather Research and Forecasting(WRF) fields were processed using CALMET to produce CALPUFF-ready meteorological inputs. Also, the distance from Ansim Briquette Fuel Complex to the residence of each environmental pneumoconiosis patient was analyzed. In addition, the affecting region of the pollutants emitted from briquette factories in Ansim Briquette Fuel Complex was determined. Results: CALPUFF modeling results showed that the highest concentrations of $PM_{10}$ were found near around the fuel complex. The modeled $PM_{10}$ distributions were characterized by significant decreases in concentration with distance from the complex. Seasonally, the highest concentration of $45{\mu}g/m^3$ was calculated in October which was mostly due to the distinct variation of amount of emission. Additional modeling with the maximum $PM_{10}$ emission of about 88 tons per year in 1986 showed that the highest concentration in October was nearly increased by 8 times than the concentration modeled with emission of 2010. As a result of medical examination and interviews for the residents in Ansim Briquette Fuel Complex and its surroundings, 8 environmental pneumoconiosis patients were found. These patients do not have occupational exposure and history. These patients have lived 0.3~1.1 km area in Ansim Briquette Fuel Complex and its surroundings. Conclusions: Airborne particles emitted from Ansim Briquette Fuel Complex can contribute to significant increase in $PM_{10}$ concentration in residential areas near around the complex. Especially, the residents near fuel complex may exposed to the pollutants emitted from the factories in Ansim Briquette Fuel Complex.

The Role of Hydroxyl Radical in the Pathogenetic Mechanism of Endotoxin-Induced Acute Lung Injury in Rats (내독소에 의한 백서의 급성 폐손상에서 Hydroxyl Radical의 병인론적 역할에 관한 연구)

  • Shim, Young-Soo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.2
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    • pp.120-130
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    • 1992
  • Background: Although there have been many studies on the pathogenetic mechanism of acute lung injury, it is still elusive. Recently interests have been focused on the role of oxygen free radicals. But the effect of hydroxyl radical on the neutrophil mobilization and the alveolar-capillary permeability is not clear especially in the endotoxin-induced acute lung injury model of rats. This investigation was performed to evaluate the pathogenetic role of hydroxyl radical on the neutrophil accumulation into the lung and the increased alveolar-capillary permeability in the endotoxin-induced acute lung injury in rats. Method: Fifty rats were divided into four groups: vehicle control group (n=5, 6hrs; n=5, 24hrs), endotoxin-treated group (n=10, 6hrs; n=10, 24hrs), Dimethylthiourea (DMTU)-pretreated group (n=10, 6hrs), and deferoxamine (DFX)-pretreated group (n=10, 6hrs). Thirty minutes before sacrifice, $^{125}I$-tagged bovine serum albumin was injected. Six and twenty four hours after endotoxin injection, the rats were sacrificed, and the radioactivity of lung tissue and peripheral blood was counted. Permeability index was defined as the ratio of radioactivity between lung tissue and peripheral blood. Another set of rats (n=52) were divided into the same four groups as before [vehicle control group (n=5, 6hrs; n=5, 24hrs), endotoxin·treated group (n=7, 6hrs; n=8, 24hrs), DMTU-pretreated group (n=6, 6hrs; n=9, 24hrs), and DFX-pretreated group (n=5, 6hrs; n=7, 24hrs)], and were sacrificed 6 and 24 hours after endotoxin injection. In these rats, cell profile of peripheral blood and bronchoalveolar lavage fluid was evaluated, and the pathologic examination of lung tissue was performed. Results: 1) Increased alveolar-capillary permeability was observed 6 hours after endotoxin injection, which was normalized after 24 hours, and this increase was attenuated by pretreatment with DMTU and DFX. 2) Neutrophil sequestration into the lung was observed 24 hours after endotoxin administration, but this was not influenced by DMTU and DFX pretreatment. Conclusion: These results suggest that hydroxyl radical would not be involved in the sequestration of neutrophils into the lung, but plays an important role in the increase of alveolar-capillary permeability in the endotoxin-induced acute lung injury in rats.

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Analysis of Interactions in Multiple Genes using IFSA(Independent Feature Subspace Analysis) (IFSA 알고리즘을 이용한 유전자 상호 관계 분석)

  • Kim, Hye-Jin;Choi, Seung-Jin;Bang, Sung-Yang
    • Journal of KIISE:Computer Systems and Theory
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    • v.33 no.3
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    • pp.157-165
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    • 2006
  • The change of external/internal factors of the cell rquires specific biological functions to maintain life. Such functions encourage particular genes to jnteract/regulate each other in multiple ways. Accordingly, we applied a linear decomposition model IFSA, which derives hidden variables, called the 'expression mode' that corresponds to the functions. To interpret gene interaction/regulation, we used a cross-correlation method given an expression mode. Linear decomposition models such as principal component analysis (PCA) and independent component analysis (ICA) were shown to be useful in analyzing high dimensional DNA microarray data, compared to clustering methods. These methods assume that gene expression is controlled by a linear combination of uncorrelated/indepdendent latent variables. However these methods have some difficulty in grouping similar patterns which are slightly time-delayed or asymmetric since only exactly matched Patterns are considered. In order to overcome this, we employ the (IFSA) method of [1] to locate phase- and shut-invariant features. Membership scoring functions play an important role to classify genes since linear decomposition models basically aim at data reduction not but at grouping data. We address a new function essential to the IFSA method. In this paper we stress that IFSA is useful in grouping functionally-related genes in the presence of time-shift and expression phase variance. Ultimately, we propose a new approach to investigate the multiple interaction information of genes.

Neuroprotective effects of resveratrol via anti-apoptosis on hypoxic-ischemic brain injury in neonatal rats (신생 백서의 저 산소 허혈 뇌손상에서 항세포사멸사를 통한 resveratrol의 신경보호 효과)

  • Shin, Jin Young;Seo, Min Ae;Choi, Eun Jin;Kim, Jin Kyung;Seo, Eok Su;Lee, Jun Hwa;Chung, Hai Lee;Kim, Woo Taek
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1102-1111
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    • 2008
  • Purpose : Resveratrol, extracted from red wine and grapes, has an anti-cancer effect, an antiinflammatory effect, and an antioxidative effect mainly in heart disease and also has neuroprotective effects in the adult animal model. No studies for neuroprotective effects during the neonatal periods have been reported. Therefore, we studied the neuroprotective effect of resveratrol on hypoxic-ischemic brain damage in neonatal rats via anti-apoptosis. Methods : Embryonic cortical neuronal cell culture of rat brain was performed using pregnant Sprague-Dawley (SD) rats at 18 days of gestation (E18) for the in vitro approach. We injured the cells with hypoxia and administered resveratrol (1, 10, and $30{\mu}g/mL$) to the cells at 30 minutes before hypoxic insults. In addition, unilateral carotid artery ligation with hypoxia was induced in 7-day-old neonatal rats for the in vivo approach. We injected resveratrol (30 mg/kg) intraperitoneally into animal models. Real-time PCR and Western blotting were performed to identify the neuroprotective effects of resveratrol through anti-apoptosis. Results : In the in vitro approach of hypoxia, the expression of Bax, caspase-3, and the ratio of Bax/Bcl-2, indicators of the level of apoptosis, were significantly increased in the hypoxia group compared to the normoxia group. In the case of the resveratrol-treated group, expression was significantly decreased compared to the hypoxia group. And the results in the in vivo approach were the same as in the in vitro approach. Conclusion : The present study demonstrates that resveratrol plays neuroprotective role in hypoxic-ischemic brain damage during neonatal periods through the mechanism of anti-apoptosis.

Ultrastructural analysis and quantification of autophagic vacuoles in wild-type and atg5 knockout mouse embryonic fibroblast cells (정상 및 atg5 유전자 제거 섬유아세포에서 자가포식체의 미세구조 및 이들의 정량적 분석)

  • Choi, Suin;Jeon, Pureum;Huh, Yang Hoon;Lee, Jin-A
    • Analytical Science and Technology
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    • v.31 no.5
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    • pp.208-218
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    • 2018
  • Autophagy is a cellular process whereby cytosolic materials or organelles are taken up in a double-membrane vesicle structure known as an autophagosome and transported into a lysosome for degradation. Although autophagy has been studied at the genetic, cellular, or biochemical level, systematic ultrastructural quantitative analysis of autophagosomes during the autophagy process by using transmission electron microscopy (TEM) has not yet been reported. In this study, we performed ultrastructural analysis of autophagosomes in wild-type (WT) mouse embryonic fibroblasts (MEFs) and autophagy essential gene (atg5) knockout (KO) MEFs. First, we performed ultrastructural analysis of autophagosomes in WT MEFs compared to atg5 KO MEFs in basal autophagy or starvation-induced autophagy. Although we observed phagopore, early, late autophagosomes, or autolysosomes in WT MEFs, atg5 KO MEFs had immature autophagosomes that showed incomplete closure. Upon starvation, late autophagosomes accumulated in WT MEFs while the number of immature autophagosomes significantly increased in atg5 KO MEF indicating that atg5 plays an important role in the maturation of autophagosomes. Next, we examined autophagosomes in the cell model expressing polyQ-expanded N-terminal fragment of huntingtin. Our TEM analysis indicates that the number of late autophagosomes was significantly increased in the cells expressing the mutant huntingtin, indicating that improving the fusion of autophagosome with lysosome may be effective to enhance autophagy for the treatment of Huntington's disease. Taken together, the results of our study indicate that ultrastructural and quantitative analysis of autophagosomes using TEM can be applied to various human cellular disease models, and that they will provide an important insight for cellular pathogenesis of human diseases associated with autophagy.