• Title/Summary/Keyword: Human immunodeficiency virus-1 (HIV-1)

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Construction of a New Gene-Fusion Expression Vector, pMONSTER

  • Baek, Chang-Ho;Wee, Sec-Han
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.663-669
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    • 2000
  • The fur (ferric uptake regulation) expression vector pMON2064 was modified to produce a Fur-fusion expression vector. A kinker site, factor Xa cleavage site, and several restriction endonuclease sites were introduced to facilitate easy cloning and isolating of the fusion protein. The resulting fusion expression vector, pMONSTER, was then used to make fusion expression vector, pMONSTER, was then used to make fusion proteins with $\beta$-galactosidase and the protease of the human immunodeficiency virus type 1 (HIV-1 PR). Strain SW4020 harboring the Fur $\beta$-galactosidase fusion vector produced blue colonies on a 5-bromo-4-chloro-3-indolyl-$\beta$-D-galactoside plate and the resulting 133 kDa fusion protein reacted with an anti-Fur antibody. The strain harboring the Fur-HIV-1 PR fusion vector produced a 29 kDa fusion protein, which also reacted with an anti-Fur antibody. The Fur-HIV-1 PR fusion protein was purified by a single column application that was designed to isolate the Fur protein. The purified Fur-HIV-1 PR fusion protein digested with factor Xa cleaved a recombinant Gag protein to release smaller fragments, including a p24 capsid protein. The Fur-HIV-1 PR fusion protein itself did not exhibit any proteolytic activity.

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Stability of Retroviral Vectors Against Ultracentrifugation Is Determined by the Viral Internal Core and Envelope Proteins Used for Pseudotyping

  • Kim, Soo-hyun;Lim, Kwang-il
    • Molecules and Cells
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    • v.40 no.5
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    • pp.339-345
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    • 2017
  • Retroviral and lentiviral vectors are mostly pseudotyped and often purified and concentrated via ultracentrifugation. In this study, we quantified and compared the stabilities of retroviral [murine leukemia virus (MLV)-based] and lentiviral [human immunodeficiency virus (HIV)-1-based] vectors pseudotyped with relatively mechanically stable envelope proteins, vesicular stomatitis virus glycoproteins (VSVGs), and the influenza virus WSN strain envelope proteins against ultracentrifugation. Lentiviral genomic and functional particles were more stable than the corresponding retroviral particles against ultracentrifugation when pseudotyped with VSVGs. However, both retroviral and lentiviral particles were unstable when pseudotyped with the influenza virus WSN strain envelope proteins. Therefore, the stabilities of pseudotyped retroviral and lentiviral vectors against ultracentrifugation process are a function of not only the type of envelope proteins, but also the type of viral internal core (MLV or HIV-1 core). In addition, the fraction of functional viral particles among genomic viral particles greatly varied at times during packaging, depending on the type of envelope proteins used for pseudotyping and the viral internal core.

Interaction of HIV-1 Core p24 Antigen with Human Monocytic Cell Line THP1 Results in TNF-${\alpha}$ Dependent Secretion of Matrix Metalloproteinase-9

  • Sung, Ji-Hye;Yoo, Seung-Hee;Park, Hae-Kyung;Chong, Young-Hae
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.9-18
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    • 2000
  • Immunological mechanisms involving the release of inflammatory factors by HIV-1 infected microglia in the brain have been implicated in the pathogenesis of HIV dementia (HIVD). Since the regulation of matrix metalloproteinases (MMPs) activity can be influenced by variety of inflammatory mediators, this study was undertaken to look for a correlation between the MMP-9 release and the production of TNF-${\alpha}$ in response to HIV-1 p24 in the human monocyte cell line THP-1 as a model for microglia. First, it was shown that HIV-l core p24 antigen induced THP-1 to secrete MMP-9 in a dose response manner while it elicited a little effect on MMP-2 release in human astroglial cell line T98G. Next, it was found that p24 induced THP-1 to secrete TNF-${\alpha}$ without prior differentiation into macrophages by phorbol myristate acetate (PMA) treatment. Furthermore, anti-TNF-${\alpha}$ neutralizing antibodies significantly blocked p24-induced MMP-9 release in a dose dependent manner. Our data indicate that p24 antigen induces monocytic MMP-9 release by triggering up-regulation of TNF-${\alpha}$ secretion.

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Knowledge and Attitude of Physical Therapist to Patient with Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome (물리치료사의 HIV/AIDS에 대한 지식과 태도)

  • Ahn So-youn;Kim Byung-jo;Kim Soo-min;Kim Jae-ho;Kim Jong-soon
    • The Journal of Korean Physical Therapy
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    • v.15 no.4
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    • pp.235-254
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    • 2003
  • The purpose of this descriptive study was to investigate the general knowledge and attitude of physical therapist to patient with HIV/AIDS. Many physical therapists are afraid of AIDS patients because they have a little information of AIDS and fear of HIV infection from AIDS patients at work site. This study was carried out from June to October in 2003 for collecting data. The 271 physical therapists volunteered for a this study and completed questionnaires. The findings of this study can be summarized as follows. 1)The knowledge of prevention of HIV/AIDS influences the attitude of physical therapists to patients with HIV/AIDS. 2)The knowledge of symptoms of HIV/AIDS influences the attitude of physical therapists to patients with HIV/AIDS. 3)The knowledge of transmission routes of HIV/AIDS influences the attitude of physical therapists to patients with HIV/AIDS. 4)84.9 percentage of participant in survey suggested college should have curriculum about HIV/AIDS. In conclusion, physical therapists with more knowledge of HIV/ADIS could positively treat HIV/AIDS patients.

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The Extracts of Kalopanax pictus Nakai. for Inhibitory Effects on HIV-1 and Its Essential Enzymes (Human Immunodeficiency Virus Type Ⅰ에 대한 음나무 추출물의 억제활성)

  • Yu Young Beob;Shim Bum Sang;Ahn Kyoo Seok;Choi Seung Hoon;Park Jong Cheol;Miyashiro H.;Hattori M.
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.4
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    • pp.1129-1133
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    • 2004
  • For the purpose of developing new anti-HIV agents from natural sources, the extracts of Kalopanax pictus were tested for their inhibitory effects on HIV-1 replication and its essential enzymes as the reverse transcriptase (RT). protease and α-glucosidase. In the assay of HIV-1-infected human T-cell line, water extracts of stem and leafstalk inhibited the HIV-1-induced cytopathic effects with Ie (inhibitory concentration) of 25 and 50㎍/㎖, respectively. Moreover water extracts (100㎍/㎖) of stem and leafstalk showed strong activity of 80% and 90% on anti-HIV-1 RT using Enzyme Linked Oligonucleotide Sorbent Assay (ELOSA) method. In the HIV-1 protease inhibition assay, aqueous stem extract inhibited the activity of the enzyme to cleave an oligopeptide, resembling one of the cleavage sites in the viral polyprotein which can only be processed by HIV-1 protease with 58%, but no glucosidase inhibitory activities. We found out this result, for these samples it is possible that the inhibition of the viral replication in vitro is due to the inhibition at least one of RT and protease. It would be of great interest to identify the compounds which are responsible for this inhibition, since all therapeutically useful agent up to date are RT, PR and α-glucosidase inhibitors.

Comparison of Analysis Methods for Detection of Replication Competent Virus and Functional Titers of HIV-l Based Lentivirus Vector (HIV-l 유래 렌티바이러스 벡터의 복제가능 바이러스 검출과 역가측정 분석방법 비교)

  • Chang Seok Kee;Oh Il Ung;Jeong Jayoung;Ahn Kwang Soo;Sohn Yeowon
    • YAKHAK HOEJI
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    • v.49 no.3
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    • pp.217-224
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    • 2005
  • Human Immunodeficiency Virus type 1 (HIV-l) based lentivirus vector has demonstrated great potential as gene therapy vectors mediating efficient gene delivery and long-term transgene expression in both dividing and nondividing cells. However, for clinical studies it must be confirmed that vector preparations are safe and not contaminated by replication competent lentivirus (RCL) related to the parental pathogenic virus, HIV-l. In this study, we would like to establish the method for titration and RCL detection of lentivirus vector. The titration was determined by vector expression containing the green fluorescent protein, GFP in transduced cells. The titer was $1{\times}10^7$ Transducing Unit/ml in the GFP expression assay and $8.9{\times}10^7$ molecules/ml in the real-time PCR. Also, for the detection of RCL, we have used a combination method of PCR and p24 antigen detection. First, PBS/psi and VSV-G region in the genomic DNA of transduced cells was detected by PCR assay. Second, transfer and expression of the HIV-1 gag gene was detected by p24 ELISA. In an attempt to amplify any RCL, the transduced cells were cultured for 3 weeks (amplification phase) and the supernatant of amplified transduced cell was used for the second transduction to determine whether a true RCL was present (indicator phase). Analysis of cells and supernatant at day 6 in indicator phase were negative for PBS/psi, VSV-G, and p24 antigen. These results suggest that they are not mobilized and therefore there are no RCL in amplification phase. Thus, real-time PCR is a reliable and sensitive method for titration and RCL detection of lentivirus vector.

A Study of Assessing Educational Needs on Program Planning for AIDS(Acquired Immunodeficiency Syndrome) Inservice Education for Clinical Nurses (임상간호사를 위한AIDS(후천성 면역 결핍증) 교육 프로그램 개발을 위한 기초 연구)

  • 박인혜;이숙자;이정희
    • Journal of Korean Academy of Nursing
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    • v.29 no.2
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    • pp.371-382
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    • 1999
  • Nurses are at the forefront of providing hands-on care to patients infected with HIV(Human Immunodeficiency Virus). Therefore, appropriate and sensible infection control precaution should be taken at all times. The purpose of this study is to find out nurse's loaming needs about HIV/AIDS ; and nurse's willingness to perform nursing care to HIV/AIDS patients ; and the degree of fellowing universal precautions in nursing care ; and the degree of knowledge of HIV/AIDS to provide information on the guide for the further continuing education. The respondents of this study were 280 nurses in a university hospital in Kwang-Ju city. Data were collected using a self-administered questionnaire and analyzed by using percentages, means, Pearson's correlation, t-test, ANOVA, Duncan's grouping. The findings were as follows ; 1. The mean age of the nurses was 28.9 years, and the mean duration of their clinical experience was 6.8 years. Majority of the nurses(83.9%) had no experience in taking care of the AIDS patients. 2. The content s nurse's need to learn were showed in order such as clinical manifestations, infection control, patient care, and the mode of transmission of HIV/AIDS. 3. Even though the nurses had quite low knowledge about HIV/AIDS(mean score was 12.2 from the total of 26), they are willing to take care of the AIDS patients (mean score was 39.5 from the total of 60), and the degree of following universal precaution was also not so bad (mean score was 40.8 from the total of 50). 4. Nurses who are over 35 years old and had informal education regarding HIV/AIDS better following universal precautions. Nurses who had over 12 years of clinical experiences have more willingness to perform nursing care to the AIDS patients. Nurses who had experiences in taking care of the AIDS patients were more knowledgeable about HIV/AIDS. These findings suggest that some strategies should be develope to increase the willingness to perform the nursing care to the AIDS patients. And also continuing education program should be develope and run for the clinical nurses so that they can provide effective and appropriate nursing care to the AIDS patients.

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Dry-Heat Treatment Process for Enhancing Viral Safety of an Antihemophilic Factor VIII Concentrate Prepared from Human Plasma

  • Kim, In-Seop;Choi, Yong-Woon;Kang, Yong;Sung, Hark-Mo;Shin, Jeong-Sup
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.997-1003
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    • 2008
  • Viral safety is a prerequisite for manufacturing clinical antihemophilic factor VIII concentrates from human plasma. With particular regard to the hepatitis A virus (HAV), a terminal dry-heat treatment ($100^{\circ}C$ for 30 min) process, following lyophilization, was developed to improve the virus safety of a solvent/detergent-treated antihemophilic factor VIII concentrate. The loss of factor VIII activity during dry-heat treatment was of about 5%. No substantial changes were observed in the physical and biochemical characteristics of the dry-heat-treated factor VIII compared with those of the factor VIII before dry-heat treatment. The dry-heat-treated factor VIII was stable for up to 24 months at $4^{\circ}C$. The dry-heat treatment after lyophilization was an effective process for inactivating viruses. The HAV, murine encephalomyocarditis virus (EMCV), and human immunodeficiency virus (HIV) were completely inactivated to below detectable levels within 10 min of the dry-heat treatment. Bovine herpes virus (BHV) and bovine viral diarrhea virus (BVDV) were potentially sensitive to the treatment. However porcine parvovirus (PPV) was slightly resistant to the treatment. The log reduction factors achieved during lyophilization and dry-heat treatment were ${\geq}5.55$ for HAV, ${\geq}5.87$ for EMCV, ${\geq}5.15$ for HIV, 6.13 for BHV, 4.46 for BVDV, and 1.90 for PPV. These results indicate that dry-heat treatment improves the virus safety of factor VIII concentrates, without destroying the activity. Moreover, the treatment represents an effective measure for the inactivation of non-lipid-enveloped viruses, in particular HAV, which is resistant to solvent/detergent treatment.

Hindsiipropane B alleviates HIV-1 Tat-induced inflammatory responses by suppressing HDAC6-NADPH oxidase-ROS axis in astrocytes

  • Jo, Hyundong;Jang, Ha Young;Youn, Gi Soo;Kim, Donggyu;Lee, Chae Yeon;Jang, Jae Hee;Choi, Soo Young;Jun, Jong-Gab;Park, Jinseu
    • BMB Reports
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    • v.51 no.8
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    • pp.394-399
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    • 2018
  • Human immunodeficiency virus-1 (HIV-1) transactivator of transcription (Tat) is an important viral factor in neuro-inflammation. Hindsiipropane B, present in Celastrus hindsii, possesses various biological mechanisms including anti-inflammatory activity. In this report, we explored the regulatory activity of hindsiipropane B on HIV-1 Tat-mediated chemokine production and its mode of action in astrocytes. Hindsiipropane B significantly alleviated HIV-1 Tat-mediated production of inflammatory chemokines, CCL2, CXCL8, and CXCL10. Hindsiipropane B inhibited expression of HDAC6, which is important regulator in HIV-1 Tat-mediated chemokine production. Hindsiipropane B diminished HIV-1 Tat-mediated reactive oxygen species (ROS) generation and NADPH oxidase activation/expression. Furthermore, hindsiipropane B inhibited HIV-1 Tat-mediated signaling cascades including MAPK, $NF-{\kappa}B$, and AP-1. These data suggest that hindsiipropane B exerts its inhibitory effects on HIV-1 Tat-mediated chemokine production via down-regulating the HDAC6-NADPH oxidaseMAPK-$NF-{\kappa}B$/AP-1 signaling axis, and could serve as a therapeutic lead compound against HIV-1 Tat-associated neuro-inflammation.

Modified SOD for Cosmeceuticals

  • Kang, Nae-Gyu;Lim, Jun-Man;Chang, Min-Youl;Park, Sun-Gyoo;Cho, Wan-Goo;Kang, She-Hoon;Park, Soo-Young
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.630-644
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    • 2003
  • A human Cu, Zn-superoxide dismutase (Cu, Zn-SOD) was fused with a Tat PTD of HIV-1 to produce a novel anti-aging ingredient, Tat-SOD for cosmeceuticals. Test of stability and evaluation of transduction efficacy and enzymatic activity suggest Tat-SOD is an effective active ingredient for anti-aging treatment.

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