• Title/Summary/Keyword: human-to-human (H2H)

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C-terminal Truncation Mutant of the Human ${\beta}_2$-adrenergic Receptor Expressed in E. coli as a Fusion Protein Retains Ligand Binding Affinity

  • Shin, Jin-Chul;Lee, Sang-Derk;Shin, Chan-Young;Lee, Sang-Bong;Ko, Kwang-Ho
    • Biomolecules & Therapeutics
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    • v.4 no.1
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    • pp.97-102
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    • 1996
  • To investigate whether human $\beta$$_2$-adrenergic receptor devoid of the C-terminal two transmembrane helices retain its ligand binding activity and specificity, 5'780-bp DNA fragment of the receptor gene which encodes amino acid 1-260 of human $\beta$$_2$-adrenergic receptor was subcloned into the bacterial fusion protein expression vector and expressed as a form of glutathione-S-transferase (GST) fusion protein in E. coli DH5$\alpha$. The receptor fusion protein was expressed as a membrane bound form which was verified by SDS-PAGE and Western blot. The fusion protein expressed in this study specifically bound $\beta$-adrenergic receptor ligand [$^3$H] Dihydroalprenolol. In saturation ligand binding assay, the $K_{d}$ value was 7.6 nM which was similar to that of intact $\beta$$_2$-adrenergic receptor in normal animal tissue ( $K_{d}$=1~2 nM) and the $B_{max}$ value was 266 fmol/mg membrane protein. In competition binding assay, the order of binding affinity of various adrenergic receptor agonists to the fusion protein was isoproterenol》epinephrine norepinephrine, which was similar to that of intact receptor in normal animal tissue. These results suggest that N-terminal five transmembrane helices of the $\beta$$_2$-adrenergic receptor be sufficient to determine the ligand binding activity and specificity, irrespective of the presence or absence of the C-terminal two transmembrane helices.s.s.s.

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Studies on the Preparation of $^{99m}TC$ Labelled Human Albumin Microsphere for Lung Scintigraphy (폐 신티그래피용 $^{99m}TC$ 표지 미소구형인 혈청 알부민의 제조에 관한 연구)

  • Park, Kyung-Bae;Awh, Ok-Doo;Kim, Jae-Rok
    • The Korean Journal of Nuclear Medicine
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    • v.23 no.1
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    • pp.63-69
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    • 1989
  • In order to develop technetium-99m-labelled human albumin microsphere (HAM) for lung scintigraphy, various experimemts such as preparation and fractionation of HAM, establishment of optimal labelling conditions, determination of radiochemical purity, stability test and biodistribution of $^{99m}Tc-HAM$ were carried out. HAM was prepared from the suspension of 1ml aqueous human serum albumin (25%) in 130 ml of olive oil at $130\sim135^{\circ}$ with vigorous stirring. The resulting HAM was fractionated with microsieve to get the desired particle size $(15\sim50\mu)$ and autoclaved for sterilizaiton. The HAM particles were treated with stannous chloride and the pH of the suspension was adjusted to $3.0\sim3.5$ with phosphate buffer. After freeze-drying the contents of single reaction vial containing 5 mg of HAM and 0.2 mg of $SnCl_2$ it was reacted with $Na^{99m}TcO_4$. The labelling yield was higher than 99.5% and the stability of $^{99m}Tc-HAM$ was high enough to maintain 99.1% of radiochemical purity up to 24 hours. Lung and liver uptake in mice was found to be 94% and 0.9%, respectively. Excellent rabbit and human lung scans were also obtained.

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Aquaporin-3 Downregulation in Vitiligo Keratinocytes Increases Oxidative Stress of Melanocytes

  • Nan-Hyung Kim;Ha Jung Kim ;Ai-Young Lee
    • Biomolecules & Therapeutics
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    • v.31 no.6
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    • pp.648-654
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    • 2023
  • Oxidative stress-induced melanocyte apoptosis is linked to the immune system and plays a critical role in the pathogenesis of vitiligo. Aquaporin-3 (AQP3), which is downregulated in vitiligo keratinocytes, regulates intracellular H2O2 accumulation. However, the role of AQP3 in oxidative stress is uncertain in vitiligo. This study investigated the effect of downregulated AQP3 on oxidative stress in vitiligo using lesional and non-lesional skin specimen sets from vitiligo patients and primary cultured adult normal human epidermal keratinocytes, with or without downregulation and overexpression of AQP3 in the presence or absence of H2O2 treatment. The levels of nuclear factor E2-related factor 2 (NRF2) and/or its main target, NAD(P)H quinone dehydrogenase 1 (NQO-1), were lower in the lesional keratinocytes and cultured keratinocytes with AQP3 knockdown, but were increased in keratinocytes upon AQP3 overexpression. Ratios of NRF2 nuclear translocation and NQO-1 expression levels were further reduced in AQP3-knockdown keratinocytes following H2O2 treatment. The conditioned media from AQP3-knockdown keratinocytes treated with H2O2 contained higher concentrations of reactive oxygen species (ROS). Moreover, the number of viable melanocytes was reduced when the conditioned media were added to the culture media. Overall, AQP3 downregulation in the keratinocytes of patients with vitiligo can induce oxidative stress in neighboring melanocytes, leading to melanocyte death.

Solid-phase Refolding of Poly-lysine Tagged Fusion Protein of hEGF and Angiogenin

  • Park Sang Joong;Ryu Kang;Suh Chang Woo;Chai Young Gyu;Kwon Oh Byung;Park Seung Kook;Lee Eun Kyu
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.1
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    • pp.1-5
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    • 2002
  • A fusion protein, consisting of a human epidermal growth factor (hEGF) as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as inclusion body in recombinant E. coli, and when the conventional, solution-phase refolding process was used the refolding yield was very low due to severe aggregation. It was probably because of the opposite electric charge at a neutral pH resulting from the vastly different pI values of each domain. The solid-phase refolding process that exploited the ionic interactions between ionic exchanger surface and the fusion protein was tried, but the adsorption yield was also very low, below $ 30\%$, regardless of the resins and pH conditions used. Therefore, to provide a higher ionic affinity toward the solid matrix, six lysine residues were tagged to the N-terminus of the hEGF domain. When heparin-Sepharose was used as the matrix, the adsorption capacity increased 2.5-3 times to about $88\%$. Besides the intrinsic affinity of angiogenin to heparin, the poly-lysine tag provided additional ionic affinity. And the subsequent refolding yield increased nearly 13-fold, from ca. $4.8\%$ in the conventional refolding of the untagged fusion protein to $63.6\%$. The process was highly reproducible. The refolded protein in the column eluate retained RNase bioactivity of angiogenin.

Extraction or gaze point on display based on EOG for general paralysis patient (전신마비 환자를 위한 EOG 기반 디스플레이 상의 응시 좌표 산출)

  • Lee, D.H.;Yu, J.H.;Kim, D.H.
    • Journal of rehabilitation welfare engineering & assistive technology
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    • v.5 no.1
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    • pp.87-93
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    • 2011
  • This paper proposes a method for extraction of the gaze point on display using EOG(Electrooculography) signal. Based on the linear property of EOG signal, the proposed method corrects scaling difference, rotation difference and origin difference between coordinate of using EOG signal and coordinate on display, without adjustment using the head movement. The performance of the proposed method was evaluated by measuring the difference between extracted gaze point and displayed circle point on the monitor with 1680*1050 resolution. Experimental results show that the average distance errors at the gaze points are 3%(56pixel) on x-axis, 4%(47pixel) on y-axis, respectively. This method can be used to human computer interface of pointing device for general paralysis patients or HCI for VR game application.

Characterization of Plasmid Encoding a Heat-labile Enterotoxin Originated from an Human Escherichia coli Strain O15: H11. (사람에서 유래한 장독성 대장균의 이열성장독소와 관련된 Plasmid 특성)

  • Rhyu, Mun-Gan;Kim, Gum-Ryong;Chang, Woo-Hyun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.47-52
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    • 1986
  • A heat-labile enterotoxin and no heat-stable enterotoxin producing($LT^+ST^-$) plasmid (110 kilobases in size) was isolated from an enterotoxigenic Escherichia coli of human strain O15:H11 and used for analysis of the $LT^+$ deoxyrionucleic acid region using recombinant DNA technology. A DNA segment containing the $LT^+$ DNA region which was one restriction endonuclease BamHl fragment(6.2 kb in size) was joind to a small multicopy plasmid, pUC9. E. coli K-12 strain, JM103 harboring the chimeric plasmid produced greater amounts of LT than did the enterotoxigenic E. coli O15:H11 strain. The BamHl fragment was further digested with various restriction endonucleases and contained no HindIll restriction site which is an essential in $LT^+ST^+$ plasmid. The detailed DNA sequencing of this $LT^+ST^-$ plasmid is required.

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New Cytotoxic Benzopyrans from the Leaves of Mallotus apelta

  • Kiem Phan Van;Dang Nguyen Hai;Bao Ha Viet;Huong Hoang Thanh;Minh Chau Van;Huong Le Mai;Lee Jung Joon;Kim Young Ho
    • Archives of Pharmacal Research
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    • v.28 no.10
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    • pp.1131-1134
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    • 2005
  • Two new benzopyrans 6-[1'-oxo-3'(R)-hydroxy-butyl]-5,7 -dimethoxy-2,2-dimethyl-2H-1-ben-zopyran (1) and 6-[1'-oxo-3'(R)-methoxy-butyl]-5,7 -dimethoxy-2,2-dimethyl-2H-1-benzopyran (2) were isolated from the leaves of Mallotus apelta Muell.-Arg., (Euphorbiaceae). Their chemical structures were elucidated by spectroscopic analyses, especially by 1D-, 2D-NMR and MS spectra. Compound 1 was found to have strong cytotoxic effect against two human cancer cell lines as human hepatocellular carcinoma (Hep-2, $IC_{50}\;:\;0.49\;{\mu}g/mL$) and rhabdosarcoma (RD, $IC_{50}\;:\;0.54\;{\mu}g/mL$), while compound 2 showed moderate activity against Hep-2 cell line ($IC_{50}\;:\;4.22\;{\mu}g/mL$) by in vitro assay.

Facial Region Extraction in an Infrared Image (적외선 영상에서의 얼굴 영역 자동 추적)

  • Shin, S.W.;Kim, K.S.;Yoon, T.H.;Han, M.H.;Kim, I.Y.
    • Proceedings of the KIEE Conference
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    • 2005.05a
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    • pp.57-59
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    • 2005
  • In our study, the automatic tracking algorithm of a human face is proposed by utilizing the thermal properties and 2nd momented geometrical feature of an infrared image. First, the facial candidates are estimated by restricting the certain range of thermal values, and the spurious blobs cleaning algorithm is applied to track the refined facial region in an infrared image.

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Epidemiologic and clinical features in children with acute lower respiratory tract infection caused by human metapneumovirus in 2006-2007 (2006-2007년 소아 급성 하기도 감염증에서 유행한 메타뉴모바이러스의 유행 및 임상 양상)

  • Park, Gwi Ok;Kim, Ji Hyun;Lee, Jae Hee;Lee, Jung Ju;Yun, Sin Weon;Lim, In Seok;Lee, Dong Keun;Choi, Eung Sang;Yoo, Byoung Hoon;Lee, Mi Kyung;Chae, Soo Ahn
    • Clinical and Experimental Pediatrics
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    • v.52 no.3
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    • pp.330-338
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    • 2009
  • Purpose : The causes of acute lower respiratory tract infection (ALRTI) are mostly attributable to viral infection, including respiratory syncytial virus (RSV), parainfluenza virus (PIV), influenza virus A/B (IFV A/B), or adenovirus (ADV). Several Korean studies reported human metapneumovirus (hMPV) as a common pathogen of ALRTI. However, studies on seasonal distribution and clinical differences relative to other viruses are insufficient, prompting us to perform this study. Methods : From November 2006 to October 2007, we tested nasopharyngeal aspiration specimens in children hospitalized with ALRTI with the multiplex reverse transcriptase-polymerase chain reaction to identify 6 kinds of common pathogen (hMPV, RSV, PIV, IFV A/B, and ADV). We analyzed positive rates and clinical features by respiratory chart review. Results : We detected 38 (8.4%) hMPV-positive cases out of 193 (41.8%) virus-positive specimens among 462 patients. HMPV infection prevailed from March to June with incidence peaking in April. HMPV-positive patients were aged 15 years (76.3%), and the ratio of boys to girls was 1.2:1. The median age was 27 months. HMPV primarily caused pneumonia (76.3 %) (P=0.018). Average hospitalization of HMPV-associated ALRTI patients was 5.8 days. In addition, they showed parahilar peribronchial infiltration (100%) on chest X-ray, normal white blood cell count (73.7%), and negative C-reactive protein (86.8 %) (P>0.05). All hMPV-positive patients recovered without complication. Conclusion : HMPV is a common pathogen of ALRTI in Korean children, especially in 1-5 year olds, from March to May. Immunocompetent children diagnosed with hMPV-associated ALRTI may have a good prognosis.

Parametric study on the impact of traffic-induced vibrations on residential structures in Istanbul, Turkey

  • A. Yesilyurt;M.R. Akram;A. Can Zulfikar;H. Alcik
    • Structural Monitoring and Maintenance
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    • v.11 no.2
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    • pp.87-100
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    • 2024
  • Traffic-induced vibrations (TIVs) possess the potential to induce structural damage in both historical and critical edifices. Recent investigations have underscored the adverse impact of TIVs within buildings, manifesting as a deleterious influence on the quality of life and operational efficiency of occupants. Consequently, these studies have dichotomized TIVs into two primary limit categories: the threshold for vibrations capable of causing structural damage and the limit values associated with human comfort. In this current research endeavor, an exhaustive analysis of peak ground acceleration (PGA), peak ground velocity (PGV), peak ground displacement (PGD), and the frequency spectrum of ground motions originating from diverse traffic sources has been conducted. Furthermore, the detrimental repercussions of these vibrations on structures, gauged through the assessment of the peak particle velocity (PPV) parameter, have been systematically evaluated. The findings of this study elucidate that TIVs within the examined structures do not attain magnitudes conducive to structural compromise; however, the levels surpassing human comfort limits are evident, attributable to specific sources and distances. Moreover, this investigation sheds light on the absence of comprehensive criteria and guidelines pertaining to the assessment of TIVs in structures within the Turkish Building Seismic Design Code 2018. It seeks to raise awareness among building constructors about the critical importance of addressing this issue, emphasizing the imperative for guidelines in mitigating the impact of TIVs on both structural integrity and human well-being.