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Induction of Peptide-specific CTL Activity and Inhibition of Tumor Growth Following Immunization with Nanoparticles Coated with Tumor Peptide-MHC-I Complexes

  • Sang-Hyun Kim;Ha-Eun Park;Seong-Un Jeong;Jun-Hyeok Moon;Young-Ran Lee;Jeong-Ki Kim;Hyunseok Kong;Chan-Su Park;Chong-Kil Lee
    • IMMUNE NETWORK
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    • v.21 no.6
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    • pp.44.1-44.15
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    • 2021
  • Tumor peptides associated with MHC class I molecules or their synthetic variants have attracted great attention for their potential use as vaccines to induce tumor-specific CTLs. However, the outcome of clinical trials of peptide-based tumor vaccines has been disappointing. There are various reasons for this lack of success, such as difficulties in delivering the peptides specifically to professional Ag-presenting cells, short peptide half-life in vivo, and limited peptide immunogenicity. We report here a novel peptide vaccination strategy that efficiently induces peptide-specific CTLs. Nanoparticles (NPs) were fabricated from a biodegradable polymer, poly(D,L-lactic-co-glycolic acid), attached to H-2Kb molecules, and then the natural peptide epitopes associated with the H-2Kb molecules were exchanged with a model tumor peptide, SIINFEKL (OVA257-268). These NPs were efficiently phagocytosed by immature dendritic cells (DCs), inducing DC maturation and activation. In addition, the DCs that phagocytosed SIINFEKL-pulsed NPs potently activated SIINFEKL-H2Kb complex-specific CD8+ T cells via cross-presentation of SIINFEKL. In vivo studies showed that intravenous administration of SIINFEKL-pulsed NPs effectively generated SIINFEKL-specific CD8+ T cells in both normal and tumor-bearing mice. Furthermore, intravenous administration of SIINFEKL-pulsed NPs into EG7.OVA tumor-bearing mice almost completely inhibited the tumor growth. These results demonstrate that vaccination with polymeric NPs coated with tumor peptide-MHC-I complexes is a novel strategy for efficient induction of tumor-specific CTLs.

An in vitro study of mesenchymal stem cell proliferation on titanium discs coated with rhTGF-β2/PLGA by electrospray (Electrospray법으로 rhTGF-β2/PLGA 복합체를 코팅한 티타늄에서의 간엽줄기세포 증식에 관한 연구)

  • Kim, Joohyung;Kim, Seong-Kyun;Heo, Seong-Joo;Koak, Jai-Young;Lee, Woo-Sung;Lee, Joo-Hee;Park, Ji-Man
    • The Journal of Korean Academy of Prosthodontics
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    • v.54 no.2
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    • pp.120-125
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    • 2016
  • Purpose: The purpose of this study is to identify the effect of mesenchymal stem cell proliferation on recombinant human transforming growth factor-beta (rhTGF-${\beta}2$) / poly (D,L-lactide-co-glycolide) (PLGA) treated titanium discs by electrospray. Materials and methods: Anodized titanium surface coated with PLGA was used for a control group to compare anodized titanium surface coated with 125 ng/ml and 500 ng/ml rhTGF-${\beta}2$ as test groups. Atomic force microscope (AFM) test was utilized to determine the difference in coating surface roughness, and field-emission scanning electron microscopy (FE-SEM) was taken to visualize even distribution of coating particles on titanium discs. The mesenchymal stem cell proliferation was tested by using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl-tetrazolium bromide) assay on 1st, 4th, 7th days. Results: According to AFM results, there was no statistically significant difference in titanium discs treated with PLGA and with rhTGF-${\beta}2$/PLGA (P>.05). MTT assay test results showed that there was statistically significant difference in mesenchymal stem cell proliferation on test groups compared to control groups at 7th day, and cell viability on discs coated with rhTGF-${\beta}2$ was significantly higher than control groups (P<.05). Conclusion: Titanium surface coated with rhTGF-${\beta}2$/PLGA shows statistically significant higher cell proliferation and the titanium surface coated with the higher concentration of rhTGF-${\beta}2$ presents faster cell growth activity.

Anti-proliferation, Cell Cycle Arrest, and Apoptosis Induced by Natural Liquiritigenin from Licorice Root in Oral Squamous Cell Carcinoma Cells (구강편평세포암종 세포에서 감초 유래 Liquiritigenin의 항증식, 세포주기 정지 및 세포사멸 유도)

  • Kwak, Ah-Won;Yoon, Goo;Chae, Jung-Il;Shim, Jung-Hyun
    • Journal of Life Science
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    • v.29 no.3
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    • pp.295-302
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    • 2019
  • Liquiritigenin (LG) is a chiral flavonoid isolated from the roots of licorice. It exhibits multiple biological activities including anti-oxidant, anti-cancer, and anti-inflammatory effects. In particular though, the anti-cancer activity of LG in oral squamous cell carcinoma has yet to be elucidated, and LG-induced apoptosis in oral squamous cell carcinoma remains poorly understood. In the present study, we tested the role of LG in inducing apoptosis in oral squamous cell carcinoma cells. LG treatment of HN22 cells resulted in a dose-dependent inhibition of cell viability as detected by a 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay. The induction of apoptosis in terms of Annexin V/7-Aminoactinomycin D staining, sub-G1 population, and multi-caspase activity were assessed with a $Muse^{TM}$ Cell Analyzer. Flow cytometric analysis revealed that LG treatment resulted in G2/M arrest in cell cycle progression and downregulation of cyclin B1 and CDC2 expression in a concentration-dependent manner. It also resulted in significant upregulation of p27. In addition, LG was seen to trigger the generation of reactive oxygen species and induce CCAAT/enhancer-binding protein homologous protein and 78-kDa glucose-regulated protein in concentration-dependent upregulation. The LG treatment of HN22 cells led to a loss of mitochondrial membrane potential (${\Delta}{\Psi}m$); it also reduced the levels of anti-apoptotic protein and increased the expression of apoptotic protease activating factor-1, cleaved poly (ADP-ribose)polymerase and Bax. Overall, our results indicate that the pro-apoptotic effects of LG in HN22 cells depend on the activation of both intrinsic and extrinsic signaling pathways. Thus, our results suggest that LG constitutes a natural compound with a potential role as an anti-tumor agent in oral squamous cell carcinoma.

Effect of Fabric Sound of Vapor Permeable Water Repellent Fabrics for Sportswear on Psychoacoustic Properties (스포츠웨어용 투습발수직물 소리가 심리음향학적 특성에 미치는 영향)

  • Lee, Jee-Hyun;Lee, Kyu-Lin;Jin, Eun-Jung;Yang, Yoon-Jung;Cho, Gil-Soo
    • Science of Emotion and Sensibility
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    • v.15 no.2
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    • pp.201-208
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    • 2012
  • The objectives of this study were to investigate the psychoacoustic properties of PTFE(Poly tetra Fluoroethylene) laminated vapor permeable water repellent fabrics which are frequently used for sportswear, to examine the relationship among fabrics' basic characteristics, mechanical properties and the psychoacoustic properties, and finally to propose the predicting model to minimize the psychoacoustic fabric sound. A total of 8 specimens' frictional sound were recorded and Zwicker's psychoacoustic parameters such as loudness(Z), sharpness(Z), roughness(Z), and fluctuation strength(Z) were calculated using the Sound Quality Program. Mechanical properties of specimens were measured by KES-FB system. Loudness(Z) of specimen D-1 was the highest, which means the rustling sound of the specimen D-1 was the most noisy. Statistically significant difference among film type was observed only in loudness(Z) for fabric sound. Based on ANOVA and post-hoc test, specimens were classified into less loud PTFE film group (groupI) and loud PTFE film group (groupII). Loudness(Z) was higher when staple yarn was used compared when filament yarn was used. According to the correlation between the mechanical properties of fabrics and loudness(Z) in groupI, the shear properties, compression properties and weight showed positive correlation with loudness(Z). According to the regression equation predicting loudness(Z) of groupI, the layer variable was chosen. In groupII, variables explaining the loudness(Z) were yarn types and shear hysteresis(2HG5).

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Development of PCR Diagnosis System for Plant Quarantine Seed-borne Wheat Streak Mosaic Virus (식물검역 종자전염 Wheat Streak Mosaic Virus의 PCR 검사시스템 개발)

  • Lee, Siwon;Kang, Eun-Ha;Chu, Yeon-Mee;Shin, Yong-Gil;Ahn, Tae-Young
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.112-117
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    • 2013
  • Wheat streak mosaic virus (WSMV), a member of the genus Tritimovirus in Potyviridae, severely impacts wheat and corn seed worldwide, but has yet to be detected in Korea, and hence, every effort should be made to prevent its introduction. To prevent WSMV from entering the country, it is necessary to prepare a specific, sensitive, simple, and fast detection method for routine application to plant quarantine procedures. For this reason, a two-step diagnosis system consisting of RT-PCR and nested PCR is being used for WSMV detection. In addition, a novel positive control was developed for use with the system. WSMV has been detected in seed sweet corn from Japan and seed wheat from USA by a two-step diagnosis system, the details of which are described in this study. After sequence analysis, similarities of 80.6 and 100.0% with other isolates were determined by BLAST. They showed the same topology, which was classified as 4 genotypes by various phylogenetic trees, using a poly protein encoding sequence amplification. In this analysis, WSMV-JSweet-corn2868 (JX845574) is classified as clade B, while WSMV-Uwheat1944-1 (KC754959) and WSMV-Uwheat1944-2 (KC754960) belong to clade D.

Patterns of Protein Leaching to Dispersion Medium during W/O/W Double Emulsion-Based Microencapsulation Processes (이중유제법에 근거한 미립자 제조 공정 중 단백질의 분산매로의 전이 양상)

  • Cho, Mi-Hyun;Choi, Soo-Kyoung;Sah, Hong-Kee
    • Journal of Pharmaceutical Investigation
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    • v.34 no.5
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    • pp.369-377
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    • 2004
  • The objective of this study was to investigate the patterns of protein leaching to an external phase during an ethyl acetate-based, double emulsion microencapsulation process. An aqueous protein solution (lactoglobulin, lysozyme, or ribonuclease; $W_1$) was emulsified in ethyl acetate containing poly-d,l-lactide-co-glycolide 75:25. The $W_1/O$ emulsion was transferred to a 0.5% polyvinyl alcohol solution saturated with ethyl acetate $(W_2)$. After the double emulsion was stirred for 5, 15, 30, or 45 min, additional 0.5% polyvinyl alcohol $(W_3)$ was quickly added into the emulsion. This so-called quenching step helped convert emulsion microdroplets into microspheres. After 2-hr stirring, microspheres were collected and dried. The degree of protein leaching to $W_2$ and/or $W_3$ phase was monitored during the microencapsulation process. In a separate, comparative experiment, the profile of protein leaching to an external phase was investigated during the conventional methylene chloride-based microencapsulation process. When ethyl acetate was used as a dispersed solvent, proteins continued diffusing to the $W_2$ phase, as stirring went on. Therefore, the timing of ethyl acetate quenching played an important role in determining the degree of protein microencapsulation efficiency. For example, when quenching was peformed after 5-min stirring of the primary $W_1/O$ emulsion, the encapsulation efficiencies of lactoglobulin and ribonuclease were $55.1{\pm}4.2\;and\;45.3{\pm}7.6%$, respectively. In contrast, when quenching was carried out in 45 min, their respective encapsulation efficiencies were $39.6{\pm}3.2\;and\;29.9{\pm}11.2%$. By sharp contrast, different results were attained with the methylene-chloride based process: up to 2 hr-stirring of the primary and double emulsions, less than 5% of a protein appeared in $W_2$. Afterwards, it started to partition from $W_1\;to\;W_2/W_3$, and such a tendency was affected by the amount of PLGA75:25 used to make microspheres. Different solvent properties (e.g., water miscibility) and their effect on microsphere hardening were to be held answerable for such marked differences observed with the two microencapsulation processes.

Effects of Feeding Extruded Soybean, Ground Canola Seed and Whole Cottonseed on Ruminal Fermentation, Performance and Milk Fatty Acid Profile in Early Lactation Dairy Cows

  • Chen, P.;Ji, P.;Li, Shengli
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.2
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    • pp.204-213
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    • 2008
  • Four ruminally cannulated Holstein cows averaging 43 days in milk (DIM) were used in a $4{\times}4$ Latin square to determine the effect of feeding extruded soybean, ground canola seed and whole cottonseed on ruminal fermentation and milk fatty acid profile. One hundred and twenty lactating Holstein cows, 58 (${\pm}31$) DIM, were assigned to four treatments in a completely randomized block design to study the effects of the three types of oilseeds on production parameters and milk fatty acid profile. The four diets were a control diet (CON) and three diets in which 10% extruded soybean (ESB), 5% ground canola seed (GCS) and 10% whole cottonseed (WCS) were included, respectively. Diets consisted of concentrate mix, corn silage and Chinese wild rye and were balanced to similar concentrations of CP, NDF and ADF. Ruminal fermentation results showed that ruminal fermentation parameters, dry matter intake and milk yield were not significantly affected by treatments. However, compared with the control, feeding cows with the three oilseed diets reduced C14:0 and C16:0 and elevated C18:0 and C18:1 concentrations in milk, and feeding ESB increased C18:2 and cis9, trans11 conjugated linoleic acid (CLA). Production results showed that feeding ESB tended to increase actual milk yield (30.85 kg/d vs. 29.29 kg/d) and significantly decreased milk fat percentage (3.53% vs. 4.06%) compared with CON. Milk protein (3.41%) and solid non-fat (13.27%) from cows fed WCS were significantly higher than from cows fed CON (3.24% and 12.63%, respectively). Milk urea N concentrations from cows fed the ESB (164.12 mg/L) and GCS (169.91 mg/L) were higher than cows fed CON (132.31 mg/L). However, intake of DM, 4% fat corrected milk, energy corrected milk, milk fat and protein yields, milk lactose percentage and yield, somatic cell count and body condition score were not affected by different treatments. The proportion of medium-chain fatty acid with 14 to 16 C units in milk was greatly decreased in cows fed ESB, GCS and WCS. Feeding ESB increased the concentration in milk of C18:1, C18:2, C18:3 and cis9, trans11-CLA content by 16.67%, 37.36%, 95.24%, 72.22%, respectively, feeding GCS improved C18:0 and C18:1 by 17.41% and 33.28%, respectively, and feeding WCS increased C18:0 by 31.01% compared with feeding CON. Both ruminal fermentation and production trial results indicated that supplementation of extruded soybean, ground canola seed and whole cottonseed could elevate the desirable poly- and monounsaturated fatty acid and decrease the medium chain fatty acid and saturated fatty acid content of milk fat without negative effects on ruminal fermentation and lactation performance.

Effects of Incompatibility on Protoplast Fusion between intra-and inter Species in Basidiomycete, Pleurotus spp. (느타리버섯의 불화합성(不和合性)이 종내(種內) 및 종간(種間) 원형질체(原形質體) 융합(融合)에 미치는 영향(影響))

  • Go, Seung-Joo;You, Chang-Hyun;Shin, Gwan-Chull
    • The Korean Journal of Mycology
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    • v.17 no.3
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    • pp.137-144
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    • 1989
  • Effects of incompatibility existing between intra-and interspecies in Pleurotus spp. on protoplast fusion, clamp formation of their fusants and fruitbody production were investigated. Protoplast fusion between intra-and interspecies of the fungus was achieved by Poly ethylene glycol treatment. The fusion frequency between intraspecies was a little higher than that of interspecies. Fusion frequency between interspecies was not correlated with their similarities based on isozyme patterns. In case of protoplast fusion between intra-and interspecies, the fusants from the compatible isolates produced normal fruit bodies, while those from the incompatible isolates did not produce clamp connections and fruit bodies except those of a few isolates presumed mutants.

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Design of a Wide-Frequency-Range, Low-Power Transceiver with Automatic Impedance-Matching Calibration for TV-White-Space Application

  • Lee, DongSoo;Lee, Juri;Park, Hyung-Gu;Choi, JinWook;Park, SangHyeon;Kim, InSeong;Pu, YoungGun;Kim, JaeYoung;Hwang, Keum Cheol;Yang, Youngoo;Seo, Munkyo;Lee, Kang-Yoon
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.16 no.1
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    • pp.126-142
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    • 2016
  • This paper presents a wide-frequency-range, low-power transceiver with an automatic impedance-matching calibration for TV-white-space (TVWS) application. The wide-range automatic impedance matching calibration (AIMC) is proposed for the Drive Amplifier (DA) and LNA. The optimal $S_{22}$ and $S_{11}$ matching capacitances are selected in the DA and LNA, respectively. Also, the Single Pole Double Throw (SPDT) switch is integrated to share the antenna and matching network between the transmitter and receiver, thereby minimizing the systemic cost. An N-path filter is proposed to reject the large interferers in the TVWS frequency band. The current-driven mixer with a 25% duty LO generator is designed to achieve the high-gain and low-noise figures; also, the frequency synthesizer is designed to generate the wide-range LO signals, and it is used to implement the FSK modulation with a programmable loop bandwidth for multi-rate communication. The TVWS transceiver is implemented in $0.13{\mu}m$, 1-poly, 6-metal CMOS technology. The die area of the transceiver is $4mm{\times}3mm$. The power consumption levels of the transmitter and receiver are 64.35 mW and 39.8 mW, respectively, when the output-power level of the transmitter is +10 dBm at a supply voltage of 3.3 V. The phase noise of the PLL output at Band 2 is -128.3 dBc/Hz with a 1 MHz offset.

Acute toxicity, Dermal and Ocular Irritation Studies of Taglisodog-eum ointment (탁리소독음 피부외용제형의 급성경피독성시험, 피부자극시험 및 안점막자극시험)

  • Lee, Jung Bok;Choi, Jae Hwan;Kim, Hee Taek;Kim, Yun Kyung;Yu, Young Beob
    • Herbal Formula Science
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    • v.24 no.4
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    • pp.289-300
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    • 2016
  • Objectives : Taglisodog-eum(TSE), a poly herbal formula, has been widely used to improve carbuncles by removing inflammation of the lymphatic channels in Traditional Korean Medicine. We previousely reported the action mechanism of TSE on experimental atopic dermatitis and the establishment of formulation for TSE ointment. In this study, we examined the toxicity test on skin and eye irritation by TSE ointment to prove the safety of Taglisodog-eum ointment in clinical use. Methods : Acute skin toxicity of the TSE ointment was evaluated in Sprague-Dawley(SD) rats. After dermal administration of TSE ointment(2,000mg/kg), body weight, mortality, and clinical signs of the rats were observed for 14days. Primary skin irritation and ocular irritation tests for TSE ointment were performed in male New Zealand White Rabbits. In dermal and ocular irritation test, body weight, mortality, clinical signs, Primary Irritation Index(P.I.I.), and The Index of Acute Ocular Irritaion(I.A.O.I.) of rabbit were observed after applying at abraded skin and eye balls with Taglisodog-eum ointment. Results : In the results of acute skin toxicity, no significant differences were found in body weight, the clinical sign and the mortality between control and TSE ointment treated group. In primary dermal irritation test, body weight, the clinical sign and the mortality were not significantly changed and Primary Irritation Index(P.I.I.) was 0.8, indicating TSE ointment as weak irritant material. In ocular irritation test, The Index of Acute Ocular Irritaion was 0.0, indicating TSE ointment as non-irritating to the eye of the rabbits. To evaluate toxicity of the TSE ointment in animal test, body weight, the clinical signs, the skin and eye irritation check were conducted; TSE ointment was considered to be weak dermal irritant in test animals. The no response of eye irritation test was observed in this experimental condition. Conclusions : According to the above toxicity test, We consider that this results is helpful for saying about the safety of TSE ointment in clinical use.