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Substantial Protective Immunity Conferred by a Combination of Brucella abortus Recombinant Proteins against Brucella abortus 544 Infection in BALB/c Mice

  • Arayan, Lauren Togonon;Huy, Tran Xuan Ngoc;Reyes, Alisha Wehdnesday Bernardo;Hop, Huynh Tan;Son, Vu Hai;Min, WonGi;Lee, Hu Jang;Kim, Suk
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.330-338
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    • 2019
  • Chronic infection with intracellular Brucella abortus (B. abortus) in livestock remains as a major problem worldwide. Thus, the search for an ideal vaccine is still ongoing. In this study, we evaluated the protective efficacy of a combination of B. abortus recombinant proteins; superoxide dismutase (rSodC), riboflavin synthase subunit beta (rRibH), nucleoside diphosphate kinase (rNdk), 50S ribosomal protein (rL7/L12) and malate dehydrogenase (rMDH), cloned and expressed into a pMal vector system and $DH5{\alpha}$, respectively, and further purified and applied intraperitoneally into BALB/c mice. After first immunization and two boosters, mice were infected intraperitoneally (IP) with $5{\times}10^4CFU$ of virulent B. abortus 544. Spleens were harvested and bacterial loads were evaluated at two weeks post-infection. Results revealed that this combination showed significant reduction in bacterial colonization in the spleen with a log protection unit of 1.31, which is comparable to the average protection conferred by the widely used live attenuated vaccine RB51. Cytokine analysis exhibited enhancement of cell-mediated immune response as IFN-${\gamma}$ is significantly elevated while IL-10, which is considered beneficial to the pathogen's survival, was reduced compared to control group. Furthermore, both titers of IgG1 and IgG2a were significantly elevated at three and four-week time points from first immunization. In summary, our in vivo data revealed that vaccination with a combination of five different proteins conferred a heightened host response to Brucella infection through cell-mediated immunity which is desirable in the control of intracellular pathogens. Thus, this combination might be considered for further improvement as a potential candidate vaccine against Brucella infection.

Anticarcinogenic Effects of Sargassum fulvellum Fractions on Several Human Cancer Cell Lines in vitro (모자반 분획물의 in vitro에서의 항발암효과)

  • 배송자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.3
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    • pp.480-486
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    • 2004
  • Despite many therapeutic advances in the understanding of the processes in carcinogenesis, overall mortality statistics are unlikely to change until there is reorientation of the concepts for the use of natural products as new anticarcinogenic agents. In this study, we investigated the anticarcinogenic activity, antioxidant and DPPH scavenging activity of Sargassum fulvellum (SF). SF was extracted with methanol, which was further fractionated into five different types: hexane (SFMH), ethylether (SFMEE), ethyl acetate (SFMEA), butanol (SFMB) and aqueous (SFMA) partition layers. We determined the cytotoxic effect of these layers on human cancer cells by MTT assay. Among various partition layers of SF, at starting concentration of 100 $\mu\textrm{g}$/mL, SFMEE showed very high cytotoxicity which were 92, 90 and 84% and kept high throughout 5 concentration levels sparsed by 100 $\mu\textrm{g}$/mL against all three human cancer cell lines: HepG2, HT-29 and HeLa. SFMEA showed a low cytotoxicity at the beginning concentration level, but as the concentration became denser, growth inhibition effect of cancer cell lines started to increase and at 500 $\mu\textrm{g}$/mL, it hit the highest, which were 91, 96 and 98% against the same three cell lines as above. We observed QR induced effect in all fraction layers of SF. SFMEE showed similar tendensy of QR induced effect as did against cytotoxicity. The QR induced effect of SFMEE on HepG2 cells at 25 $\mu\textrm{g}$/mL concentration indicated 3 times higher than the control value of 1.0 and SFMH tended to be concentration-dependent on HepG2 cells. At 100 $\mu\textrm{g}$/mL, the QR induced effects resulted a ratio, which was 2.5 times higher than the control value. In search for antioxidation effects of SF extract and partition layer, the reducing activity on the 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging potential was sequentially screened. The SFM has similar antioxidant activity as to BHT and vitamin C groups.

Identification of Differentially Expressed Genes Between Somatic Cell Nuclear Transfer and Normal Placenta in Cattle (소의 체세포핵이식태반과 정상태반간의 차등 발현 유전자 분석)

  • Yu, Seong-Lan;Jeong, Hang-Jin;Sang, Byung-Chan;Ryoo, Seung-Heui;Jung, Kie-Chul;Yoon, Jong-Taek;Seong, Hwan-Hoo;Jin, Dong-Il;Lee, Jun-Heon
    • Journal of Animal Science and Technology
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    • v.50 no.5
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    • pp.641-648
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    • 2008
  • There has been great success for making transgenic animals using somatic cell nuclear transfer(SCNT) up to this time. However, the success rates of the production of live transgenic animals are still very low. The current research has been carried out for delineation of differentially expressed genes between SCNT and normal placenta in cattle. In the present observations, high expression has been observed for CTSZ, LOC509426 and ELF1 genes in normal placenta. On the other hand, TIMP2, PAG1B, PAG-21, LOC782894, SERPINB6 and mKIAA2025 protein were highly expressed in SCNT placenta. Five genes, which were highly expressed in SCNT placenta, have been further investigated using semi-quantitative real-time PCR. The results were similar to that we observed using ACP. In the future, all genes affecting the SCNT and normal placenta have to be discovered and their networks will be fully investigated. The genes were identified in this study would be great help for identifying differential gene expressions in SCNT placenta.

A Comparative Study of Korean Home Economic Curriculum and American Practical Problem Focused Family & Consumer Sciences Curricula (우리나라 가정과 교육과정과 미국의 실천적 문제 중심 교육과정과의 비교고찰)

  • Kim, Hyun-Sook;Yoo, Tae-Myung
    • Journal of Korean Home Economics Education Association
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    • v.19 no.4
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    • pp.91-117
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    • 2007
  • This study was to compare the contents and practical problems addressed, the process of teaching-learning method, and evaluation method of Korean Home Economics curriculum and of the Oregon and Ohio's Practical Problem Focused Family & Consumer Sciences Curricula. The results are as follows. First, contents of Korean curriculum are organized by major sub-concepts of Home Economics academic discipline whereas curricular of both Oregon and Ohio states are organized by practical problems. Oregon uses the practical problems which integrate multi-subjects and Ohio uses ones which are good for the contents of the module by integrating concerns or interests which are lower or detailed level (related interests). Since it differentiates interest and module and used them based on the basic concept of Family and Consumer Science, Ohio's approach could be easier for Korean teachers and students to adopt. Second, the teaching-learning process in Korean home economics classroom is mostly teacher-centered which hinders students to develop higher order thinking skills. It is recommended to use student-centered learning activities. State of Oregon and Ohio's teaching-learning process brings up the ability of problem-solving by letting students clearly analyze practical problems proposed, solve problems by themselves through group discussions and various activities, and apply what they learn to other problems. Third, Korean evaluation system is heavily rely on summative evaluation such as written tests. It is highly recommended to facilitate various performance assessment tools. Since state of Oregon and Ohio both use practical problems, they evaluate students mainly based on their activity rather than written tests. The tools for evaluation include project documents, reports of learning activity, self-evaluation, evaluation of discussion activity, peer evaluation in a group for each students for their performance, assessment about module, and written tests as well.

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The comparative of Naringin and Chitosan using Natural preservation agents by LM and TEM (천연보존제 나린진과 키토산의 비교....LM & TEM적 소견)

  • Kim, In-Suk;Yoo, Geun-Chang;Chae, Soo-Chul;Lee, Chong-Bin;Jeon, Chang-Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.10 no.4
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    • pp.283-292
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    • 2005
  • This study investigated the effects of naringin, and chitosan in rabbits' corneas. Naringin, a glycone of naringenin, is a widely distributed bioflavonoid in the grapefruit and citrus peel, and it has already been reported as an antioxidant, antimicrobial, and anticancer agent. It has been used as a food preservatives and cosmetics. One of the natural preservatives, chitosan has also used in food preservatives, health drinks, and teas. Chitosan is distributed in the epithelium of crustacea, insects, and fungi. Naringin and chitosan have no harmful effects of cytotoxicity in the human body and they are recognized as an antibacterial for various forms of bacteria. The purpose of this study is to search for the ideal percentage of natural products to substitute the chemical preservatives occuring within the cornea and conjunctiva cytotoxicity and inflammations as wearing on soft contact lens. The present study compared the morphology of corneal epithelium and endothelium observed by light microscopy (LM) and transmission electron microscopy (TEM). In vivo methods, We investigated the effects of natural preservatives on soft contact lens. We inserted 3-4 drops of the naringin and chitosan, directly on rabbits' corneas 4 times per a day during one week. After enucleation of cornea, morphorgical damages of the epithelium and endothelium were observed by LM and TEM. In view of ultrastructure, chitosan caused siginficant damage on the epithelium and endothelium of cornea. The damage of cells was higher in chitosan treated cornea than 0.01, 0.1, and 1% of naringin. The 1% of naringin also expressed cell damage seriously. The results suggest that the most important thing is to use the reasonable percentage of preservatives for contact lens solutions.

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A Design of Fractional Motion Estimation Engine with 4×4 Block Unit of Interpolator & SAD Tree for 8K UHD H.264/AVC Encoder (8K UHD(7680×4320) H.264/AVC 부호화기를 위한 4×4블럭단위 보간 필터 및 SAD트리 기반 부화소 움직임 추정 엔진 설계)

  • Lee, Kyung-Ho;Kong, Jin-Hyeung
    • Journal of the Institute of Electronics and Information Engineers
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    • v.50 no.6
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    • pp.145-155
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    • 2013
  • In this paper, we proposed a $4{\times}4$ block parallel architecture of interpolation for high-performance H.264/AVC Fractional Motion Estimation in 8K UHD($7680{\times}4320$) video real time processing. To improve throughput, we design $4{\times}4$ block parallel interpolation. For supplying the $10{\times}10$ reference data for interpolation, we design 2D cache buffer which consists of the $10{\times}10$ memory arrays. We minimize redundant storage of the reference pixel by applying the Search Area Stripe Reuse scheme(SASR), and implement high-speed plane interpolator with 3-stage pipeline(Horizontal Vertical 1/2 interpolation, Diagonal 1/2 interpolation, 1/4 interpolation). The proposed architecture was simulated in 0.13um standard cell library. The gate count is 436.5Kgates. The proposed H.264/AVC Fractional Motion Estimation can support 8K UHD at 30 frames per second by running at 187MHz.

Anti-tumor effect of new compound, 127, through the induction of apoptosis (새로운 화합물 c-127의 세포고사 유도에 의한 항암효과)

  • Baek, Ki Hwan;Han, A Lum;Shin, Sae Ron;Jin, Chun Mae;Yoon, Young Wook;Yu, Seung Taek;Kim, Jong Duk;Choi, Du Young
    • Clinical and Experimental Pediatrics
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    • v.52 no.6
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    • pp.696-700
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    • 2009
  • Purpose : We screened more than 350 compounds with an endoperoxide ring structure in search of an anti-leukemic drug and found that compound 127 (c-127) could induce significant cytotoxicity in HL-60 cells. In this study, we investigated the molecular mechanisms of compound 127-induced antitumor activity on HL-60 cells. Methods : HL-60 cells were cultured in Rosewell Park Memorial Institute 1640 and cell viability was measured by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide], a tetrazole assay. Apoptosis was assessed by a DNA fragmentation test. Apoptotic machineries were determined by Western blot analysis. Results : C-127 could induce a cytotoxic effect at 24 h and apoptosis at 6 h, which was demonstrated with MTT assay and DNA fragmentation test, respectively. The apoptotic effect of this drug was caused by the activation of the intracellular caspase-8,3 activation, the cleavage of pro-apoptotic Bid, and the increase of c-Jun expression accompanied with JNK (Jun N-terminal kinases) phosphorylation. On the contrary, it increased the expression of anti-apoptotic Bcl-2 levels, leading to the induction of the induction of anti-apoptotic effect. Taken together, the present study demonstrated that c-127 was a potent inducer of cytotoxicity on HL-60 cells through apoptotic mechanisms, which included the activation of caspase family, the regulation of Bcl-2 family, and the activation of JNK signaling pathway. Conclusion : Our results suggest that c-127 has a strong antitumor activity through the regulation of various apoptotic machineries on HL-60 cells. The compound may be utilized as an effective and potentially therapeutic drug in leukemia.

Beyond Clot Dissolution; Role of Tissue Plasminogen Activator in Central Nervous System

  • Kim, Ji-Woon;Lee, Soon-Young;Joo, So-Hyun;Song, Mi-Ryoung;Shin, Chan-Young
    • Biomolecules & Therapeutics
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    • v.15 no.1
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    • pp.16-26
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    • 2007
  • Tissue plasminogen activator (tPA) is a serine protease catalyzing the proteolytic conversion of plasminogen into plasmin, which is involved in thrombolysis. During last two decades, the role of tPA in brain physiology and pathology has been extensively investigated. tPA is expressed in brain regions such as cortex, hippocampus, amygdala and cerebellum, and major neural cell types such as neuron, astrocyte, microglia and endothelial cells express tPA in basal status. After strong neural stimulation such as seizure, tPA behaves as an immediate early gene increasing the expression level within an hour. Neural activity and/or postsynaptic stimulation increased the release of tPA from axonal terminal and presumably from dendritic compartment. Neuronal tPA regulates plastic changes in neuronal function and structure mediating key neurologic processes such as visual cortex plasticity, seizure spreading, cerebellar motor learning, long term potentiation and addictive or withdrawal behavior after morphine discontinuance. In addition to these physiological roles, tPA mediates excitotoxicity leading to the neurodegeneration in several pathological conditions including ischemic stroke. Increasing amount of evidence also suggest the role of tPA in neurodegenerative diseases such as Alzheimer's disease and multiple sclerosis even though beneficial effects was also reported in case of Alzheimer's disease based on the observation of tPA-induced degradation of $A{\beta}$ aggregates. Target proteins of tPA action include extracellular matrix protein laminin, proteoglycans and NMDA receptor. In addition, several receptors (or binding partners) for tPA has been reported such as low-density lipoprotein receptor-related protein (LRP) and annexin II, even though intracellular signaling mechanism underlying tPA action is not clear yet. Interestingly, the action of tPA comprises both proteolytic and non-proteolytic mechanism. In case of microglial activation, tPA showed non-proteolytic cytokine-like function. The search for exact target proteins and receptor molecules for tPA along with the identification of the mechanism regulating tPA expression and release in the nervous system will enable us to better understand several key neurological processes like teaming and memory as well as to obtain therapeutic tools against neurodegenerative diseases.

A Simple Dosing Scheme for Intravenous Busulfan Based on Retrospective Population Pharmacokinetic Analysis in Korean Patients

  • Choe, Sang-Min;Kim, Gayeong;Lim, Hyeong-Seok;Cho, Sang-Heon;Ghim, Jong-Lyul;Jung, Jin-Ah;Kim, Un-Jib;Noh, Gyu-Jeong;Bae, Kyun-Seop;Lee, Dong-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.4
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    • pp.273-279
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    • 2012
  • Busulfan is an antineoplastic agent with a narrow therapeutic window. A post-hoc population pharmacokinetic analysis of a prospective randomized trial for comparison of four-times daily versus once-daily intravenous busulfan was carried out to search for predictive factors of intravenous busulfan (iBu) pharmacokinetics (PK). In this study the population PK of iBu was characterized to provide suitable dosing recommendations. Patients were randomized to receive iBu, either as 0.8 mg/kg every 6 h or 3.2 mg/kg daily over 4 days prior to hematopoietic stem cell transplantation. In total, 295 busulfan concentrations were analyzed with NONMEM. Actual body weight and sex were significant covariates affecting the PK of iBu. Sixty patients were included in the study (all Korean; 23 women, 37 men; mean [SD] age, 36.5 [10.9] years; weight, 66.5 [11.3] kg). Population estimates for a typical patient weighing 65 kg were: clearance (CL) 7.6 l/h and volume of distribution (Vd) 32.2 l for men and 29.1 L for women. Inter-individual random variabilities of CL and $V_d$ were 16% and 9%. Based on a CL estimate from the final PK model, a simple dosage scheme to achieve the target $AUC_{0-inf}$ (defined as median AUC0-inf with a once-daily dosage) of 26.18 $mg/l{\cdot}hr$, was proposed: $24.79{\cdot}ABW^{0.5}mg$ q24h, where ABW represents the actual body weight in kilograms. The dosing scheme reduced the unexplained interindividual variabilities of CL and Vd of iBu with ABW being a significant covariate affecting clearance of iBU. We propose a new simple dosing scheme for iBu based only on ABW.

Modeling of Extrusion for Pectin Extraction from Apple Pomace (사과박의 펙틴 추출을 위한 압출 공정 모형화)

  • Cho, Yong-Jin;Kim, Chong-Tai;Kim, Chul-Jin;Hwang, Jae-Kwan
    • Korean Journal of Food Science and Technology
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    • v.31 no.4
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    • pp.1011-1016
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    • 1999
  • This study was performed to search a physical method having high yield and quality and minimum environmental pollution for extraction of pectin from apple pomace. Based on the physical solubilization of plant cell wall under the condition of high temperature, pressure and shearing stress, apple pomace was treated by a corotating intermeshing type twin-screw extruder with the diameter-to-length ratio of 1/20. The specific mechanical energy of extruder was introduced as system parameter for extrusion process modeling and the shaft speed, feed rate and moisture content as process variables. The yield, average molecular weight and galacturonic acid content of water-soluble polysaccharides obtained by extrusion were, respectively, modeled with the linear functions of the system parameter which was of the form as a linear function of process variables. The specific mechanical energy increased with increase of shaft speed and with decrease of feed rate and moisture content. Out of process variables, moisture content had the greatest effect on specific mechanical energy. The yield increased with increase of specific mechanical energy while the average molecular weight and galacturonic acid content increased with its decrease. In aspects of yield and quality of pectin, the results from this study showed the possibility to replace a traditional acidic method with the extrusion treatment of this study.

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