• Title/Summary/Keyword: gag

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Nuclear Localization Signals in Prototype Foamy Viral Integrase for Successive Infection and Replication in Dividing Cells

  • Hossain, Md. Alamgir;Ali, Md. Khadem;Shin, Cha-Gyun
    • Molecules and Cells
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    • v.37 no.2
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    • pp.140-148
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    • 2014
  • We identified four basic amino acid residues as nuclear localization signals (NLS) in the C-terminal domain of the prototype foamy viral (PFV) integrase (IN) protein that were essential for viral replication. We constructed seven point mutants in the C-terminal domain by changing the lysine and arginine at residues 305, 308, 313, 315, 318, 324, and 329 to threonine or proline, respectively, to identify residues conferring NLS activity. Our results showed that mutation of these residues had no effect on expression assembly, release of viral particles, or in vitro recombinant IN enzymatic activity. However, mutations at residues 305 (R ${\rightarrow}$ T), 313(R ${\rightarrow}$ T), 315(R ${\rightarrow}$ P), and 329(R ${\rightarrow}$ T) lead to the production of defective viral particles with loss of infectivity, whereas non-defective mutations at residues 308(R ${\rightarrow}$ T), 318(K ${\rightarrow}$ T), and 324(K ${\rightarrow}$ T) did not show any adverse effects on subsequent production or release of viral particles. Sub-cellular fractionation and immunostaining for viral protein PFV-IN and PFV-Gag localization revealed predominant cytoplasmic localization of PFV-IN in defective mutants, whereas cytoplasmic and nuclear localization of PFV-IN was observed in wild type and non-defective mutants. However sub-cellular localization of PFV-Gag resulted in predominant nuclear localization and less presence in the cytoplasm of the wild type and non-defective mutants. But defective mutants showed only nuclear localization of Gag. Therefore, we postulate that four basic arginine residues at 305, 313, 315 and 329 confer the karyoplilic properties of PFV-IN and are essential for successful viral integration and replication.

The Comic Expressed Comedy Costume in TV - Focused on - (골계미가 표현된 TV 코미디 의상 연구 - <개그콘서트>를 중심으로 -)

  • Lee, Min-Jung;Kim, Min-Ja
    • Journal of the Korean Society of Costume
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    • v.58 no.1
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    • pp.61-78
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    • 2008
  • Comedy costumes worn on the comedians/gag men express the information of the performances such as character's era, place, social rank, present environment, age, sex, occupation, emotion, relationships between the characters, importance and mood in visual language. The comic is found when these informations are reversed, revealed, exaggerated or distorted. To analyse the TV comedy costumes, 5 subordinate concepts of the comic which are Body, Gender, Age, T.P.O.(Time, Place, Occasion) and Role could be identified, and the results from the analysis focused on (10 shows were selected from each of the first and second half of the years from 2003 to 2007) are as follows: Distorted and ugly body implies the resistance against the ideal body. Reversed or confused sex are usually expressed as men dressed in women, and these mean breaking the dichotomy between male and female, and coexistence of the masculinity and feminity, and satirizing the social custom restricting women by moral rules. It could be recognized that the way of men's dressing in women have been changing keeping pace with the times. The discord between age and costume was often expressed with children's wear and childish props. This implies the liberation from the age role. The comic expressed from the inadequate costume for T.P.O. usually appeared with the costumes 20-30years behind the times. When there were discordance with the acts and acts expected from the outfit(appearance) also made an ironical laugh. The comics acquired by vulgarization and exaggeration of the characteristics of role(figures/occupation) were from the imitation and deformation of the objects in stereotypes, and through this dissolving the custom was under way.

Detection of BLV Proviral DNA in Korean Native Goats Experimentally Infected with Bovine Leukemia Virus by Polymerase Chain Reaction (소백혈병 바이러스 (Bovine Leukemia Virus)에 감염된 한국 재래산양에서 PCR기법을 이용한 BLV 유전자 검출)

  • Jun, Moo-Hyung;Chang, Kyung-Soo;Cho, Young-Sung;Park, Jong-Hyeon;An, Soo-Hwan
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.217-225
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    • 1997
  • PCR amplication using the primers for gag, pol and env genes in BLV (bovine leukemia virus) proviral DNA and syncytium assay were carried out for the Korean native goats experimentally infected with bovine leukemia virus to investigate pathogenesis of BLV in the goats, and to establish a model animal for BLV infection. The oligonucleotide primers used in PCR revealed very high specificity. The minimal amount of FLK-BLV cellular chromosomal DNA to detect the integrated BLV proviral DNA was 10 ng. The peripheral blood lymphocytes from the goat infected with BLV were examined at regular intervals by PCR amplification and syncytium assay. Pol or gag genes were detected in none of three infected goats at the 1st week post-infection (p.i.). At the 4th week p.i., one of three goats showed the amplified gag gene. Thereafter detection rates for the genes were increased, indicating that the BLV proviral genes were integrated in all of the lymphocytes from three goats, at the 16th weeks p.i., when it was evident in syncytium assay that the lymphocytes from all of three goats were infested with infective BLV. Investigating the tissues from the necropsied goats at the 8th month p.i., the amplified BLV proviral genes and infective BLV were detected in all of the peripheral lymphocytes from three infected-goats. Among various tissues examined, the amplified BLV proviral genes were observed in spleen and superficial cervical, mandibular and retropharyngeal lymph nodes, and the infective BLV, in superficial cervical and mandibular lymph nodes. It was assumed that the Korean native goat was quite susceptible to BLV infection, indicating that the goat could be a good model animal for BLV.

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Effect of Cinnamomum Cassia on Cartilage Protection in Rabbit and Human Articular Cartilage

  • Baek, Yong-Hyeon;Huh, Jeong-Eun;Lee, Jae-Dong;Choi, Do-Young;Park, Dong-Suk
    • The Journal of Korean Medicine
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    • v.28 no.4
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    • pp.148-157
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    • 2007
  • Background & Objective: Articular cartilage is a potential target for drugs designed to inhibit the activity of matrix metalloproteinases (MMPs) to stop or slow the destruction of the proteoglycanand collagen in the cartilage extracellular matrix. The purpose of this study was to investigate the effects of Cinnamomum cassia in inhibiting the release of glycosaminoglycan (GAG), the degradation of collagen, and MMP activity in rabbit and human articular cartilage explants. Methods: The cartilage-protective effects of Cinnamomum cassia were evaluated by using glycosaminoglycan degradation assay, collagen degradation assay, colorimetric analysis of MMP activity, measurement of lactate dehydrogenase activity and histological analysis in rabbit cartilage explants culture. Results: Interleukin-1a (IL-1a) rapidly induced GAG, but collagen was much less readily released from cartilage explants. Cinnamomum cassia significantly inhibited GAG and collagen release in a concentration-dependent manner. Cinnamomum cassia dose-dependently inhibited MMP-1, MMP-3 and MMP-13 activities from IL-1a-treated cartilage explants culture when tested at concentrations ranging from 0.02 to 1 mg/ml. Conclusion : These results indicate that Cinnamomum cassia inhibits the degradation of proteoglycan and collagen through the down regulation of MMP-1, MMP-3 and MMP-13 activities of IL-1a-stimulated rabbit and human articular cartilage explants.

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Effect of Angelica gigas extract powder on progress of osteoarthritis induced by monosodium iodoacetate in rats (참당귀 추출분말이 Monosodium Iodoacetate로 유발된 흰쥐의 골관절염에 대한 효과)

  • Kwon, Jin-Hwan;Han, Min-Seok;Lee, Bu-Min;Lee, Yong-Moon
    • Analytical Science and Technology
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    • v.28 no.1
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    • pp.72-77
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    • 2015
  • To study the efficacy of extract powder of Angelica gigas in preventing and treating degeneration of the articular cartilage in rats with monosodium iodoacetate (MIA)-induced osteoarthritis, A total of 30 six-week-old Sprague-Dawley rats were randomly divided into normal control group, untreated group and Angelica gigas treated group, with 10 rats in each group. During the treatment period, body weight were measured in each four days interval from starting date. The rat were sacrificed at the end of 3rd week after daily administration of Angelica gigas and then rat tibia articular cartilage was removed. In articular cartilages, glycosaminoglycan (GAG) amount increased by MIA treatment were reduced while proteoglycan (PG) amount decreased by MIA treatment were fairly recovered by Angelica gigas treatment, respectively. The content of TNF-a was also slightly reduced sections of the cartilage were stained with safranin-0 were also partially recovered by Angelica gigas treatment. By HPLC analysis, the content of main compounds decursin and decursinol angelate was analyzed as $10.5{\pm}0.2%$ of total extracts.

Extraction of Glycosaminoglycans from Styela clava Tunic (미더덕 껍질로부터 Glycosaminoglycans의 추출)

  • 안삼환;정성훈;강석중;정태성;최병대
    • KSBB Journal
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    • v.18 no.3
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    • pp.180-185
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    • 2003
  • Glycosaminoglycans (GAGS was extracted from sea squirt, Styela clava with sodium phosphate at 105$^{\circ}C$ for 2 hr and deproteinized with trichloroacetic acid or hydrochloride. The GAGs obtained from tunic consist 41.7% crude carbohydrates, 31.8% crude protein, and 31.2% sulfate. It was mainly constituted of galactose, glucosamine, glucose, mannose, and glacrosamine. The prominent amino acid were phenylalanine, threonine, glutamic acid, and aspartic acid. Mineral contents was mainly constituted 3.0 mg% sodium, 1.6 mg% potassium, and 1.2 mg% phosphorus. Trichloroacetic acid, hydrochloride and 5-sulfosalicylic acid were used for deprotein of the GAGs. Effective volume for deprotein of crude GAGs were 5.0% trichloroacetic acid (w/v) and 10.0% HCI (v/v) treatment. The deproteinized GAGs contained 35.1%, 35.4% of protein and 22.0%, 18.5% of sulfate, respectively.

Molecular Characterization of the HERV-W Env Gene in Humans and Primates: Expression, FISH, Phylogeny, and Evolution

  • Kim, Heui-Soo;Kim, Dae-Soo;Huh, Jae-Won;Ahn, Kung;Yi, Joo-Mi;Lee, Ja-Rang;Hirai, Hirohisa
    • Molecules and Cells
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    • v.26 no.1
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    • pp.53-60
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    • 2008
  • We characterized the human endogenous retrovirus (HERV-W) family in humans and primates. In silico expression data indicated that 22 complete HERV-W families from human chromosomes 1-3, 5-8, 10-12, 15, 19, and X are randomly expressed in various tissues. Quantitative real-time RT-PCR analysis of the HERV-W env gene derived from human chromosome 7q21.2 indicated predominant expression in the human placenta. Several copies of repeat sequences (SINE, LINE, LTR, simple repeat) were detected within the complete or processed pseudo HERV-W of the human, chimpanzee, and rhesus monkey. Compared to other regions (5'LTR, Gag, Gag-Pol, Env, 3'LTR), the repeat family has been mainly integrated into the region spanning the 5'LTRs of Gag (1398 bp) and Pol (3242 bp). FISH detected the HERV-W probe (fosWE1) derived from a gorilla fosmid library in the metaphase chromosomes of all primates (five hominoids, three Old World monkeys, two New World monkeys, and one prosimian), but not in Tupaia. This finding was supported by molecular clock and phylogeny data using the divergence values of the complete HERV-W LTR elements. The data suggested that the HERV-W family was integrated into the primate genome approximately 63 million years (Myr) ago, and evolved independently during the course of primate radiation.

Glycosaminoglycan Degradation-Inhibitory Lactic Acid Bacteria Ameliorate 2,4,6-Trinitrobenzenesulfonic Acid-Induced Colitis in Mice

  • Lee, Bo-Mi;Lee, Jung-Hee;Lee, Hye-Sung;Bae, Eun-Ah;Huh, Chul-Sung;Ahn, Young-Tae;Kim, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.6
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    • pp.616-621
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    • 2009
  • To evaluate the effects of lactic acid bacteria (LAB) in inflammatory bowel diseases (IBD), we measured the inhibitory effect of several LAB isolated from intestinal microflora and commercial probiotics against the glycosaminoglycan (GAG) degradation by intestinal bacteria. Bifidobacterium longum HY8004 and Lactobacillus plantarum AK8-4 exhibited the most potent inhibition. These LAB inhibited colon shortening and myeloperoxidase production in 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced experimental colitic mice. These LAB also blocked the expression of the proinflammatory cytokines, IL-$1{\beta}$ and TNF-$\alpha$, as well as of COX-2, in the colon. LAB also blocked activation of the transcription factor, NF-${\kappa}B$, and expression of TLR-4 induced by TNBS. In addition, LAB reduced the TNBS-induced bacterial degradation activities of chondroitin sulfate and hyaluronic acid. These findings suggest that GAG degradation-inhibitory LAB may improve colitis by inhibiting inflammatory cytokine expression via TLR-4-linked NF-${\kappa}B$ activation and by inhibiting intestinal bacterial GAG degradation.

Effects of Aralia cordata Thunb. on Proteoglycan Release, Type II Collagen Degradation and Matrix Metalloproteinase Activity in Rabbit Articular Cartilage Explants

  • Baek, Yong-Hyeon;Seo, Byung-Kwan;Lee, Jae-Dong;Huh, Jeong-Eun;Yang, Ha-Ru;Cho, Eun-Mi;Choi, Do-Young;Kim, Deog-Yoon;Cho, Yoon-Je;Kim, Kang-Il;Park, Dong-Suk
    • Journal of Acupuncture Research
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    • v.22 no.2
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    • pp.191-201
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    • 2005
  • Background & Objective: Articular cartilage is a potential target for drugs designed to inhibit the activity of matrix metalloproteinases (MMPs) to stop or slow the destruction of the proteoglycan and collagen in the cartilage extracellular matrix. The purpose of this study was to investigate the effects of Aralia cordata Thunb. in inhibiting the release of glycosaminoglycan (GAG), the degradation of collagen, and MMP activity in rabbit articular cartilage explants. Methods : The cartilage-protective effects of Aralia cordata Thunb. were evaluated by using glycosaminoglycan degradation assay, collagen degradation assay, colorimetric analysis of MMP activity, measurement of lactate dehydrogenase activity and histological analysis in rabbit cartilage explants culture. Results : Interleukin-la (IL-1a) rapidly induced GAG, but collagen was much less readily released from cartilage explants. Aralia cordata Thunb. significantly inhibited GAG and collagen release in a concentration-dependent manner. Aralia cordata Thunb. dose-dependently inhibited MMP-3 and MMP-13 expression and activities from IL-1a-treated cartilage explants cultures when tested at concentrations ranging from 0.02 to 0.2 mg/ml. Aralia cordata Thunb. had no harmful effect on chondrocytes viability or cartilage morphology in cartilage explants. Histological analysis indicated that Aralia cordata Thunb. reduced the degradation of the cartilage matrix compared with that of IL -1a-treated cartilage explants.

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Comparative Analysis of Platelet-rich Plasma Effect on Tenocytes from Normal Human Rotator Cuff Tendon and Human Rotator Cuff Tendon with Degenerative Tears

  • Yoon, Jeong Yong;Lee, Seung Yeon;Shin, Sue;Yoon, Kang Sup;Jo, Chris Hyunchul
    • Clinics in Shoulder and Elbow
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    • v.21 no.1
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    • pp.3-14
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    • 2018
  • Background: Platelet-rich plasma (PRP) stimulates cell proliferation and enhances matrix gene expression and synthesis. However, there have been no comparative study of the PRP effect on the normal and degenerative tenocytes. The purpose of this study was to compare the effect of PRP on tenocytes from normal and degenerative tendon. Methods: Tendon tissues were obtained from patients undergoing arthroscopic repair (n=9) and from healthy donors (n=3). Tenocytes were cultured with 10% (vol/vol) platelet-poor plasma, PRP activated with calcium, and PRP activated with calcium and thrombin. The total cell number was assessed at days 7 and 14. The expressions of type I and III collagen, decorin, tenascin-C, and scleraxis were evaluated by quantitative real-time reverse transcriptase polymerase chain reaction. The total collagen and glycosaminoglycan (GAG) synthesis was evaluated at days 7 and 14. Results: No differences were observed between the groups at day 7, but cell proliferation was remarkably increased in tenocytes from the degenerative tendon at day 14. In both tenocyte groups, the gene expressions of type I and III collagen were up-regulated. GAG synthesis was greater in the normal tendon, whereas the expressions of decorin and tenascin-C were increased in tenocytes from the degenerative tendon. Tenocytes from the degenerative tendon had higher fold-change of GAG synthesis and a lower collagen III/I ratio than normal tenocytes. Conclusions: PRP promoted the cell proliferation and enhanced the synthesis of tendon matrix in both groups. PRP has a greater positive effect on cell proliferation, matrix gene expression and synthesis in tenocytes from degenerative tendon.