• Title/Summary/Keyword: affinity pattern

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Characterization of Humanized Antibody Produced by Apoptosis-Resistant CHO Cells under Sodium Butyrate-Induced Condition

  • Kim, No-Soo;Chang, Kern-Hee;Chung, Bo-Sup;Kim, Sung-Hyun;Kim, Jung-Hoe;Lee, Gyun-min
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.926-936
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    • 2003
  • Overexpression of human Bcl-2 protein in recombinant Chinese hamster ovary (rCHO) cells producing humanized antibody (SH2-0.32) considerably suppressed sodium butyrate (NaBu)-induced apoptosis during batch culture by using commercially available serum-free medium, which extended the culture longevity. Due to the extended culture longevity provided by the anti-apoptotic effect of Bcl-2 overexpression, the final antibody concentration of 14C6-bcl-2 culture (Bcl-2 high producer, $23\;\mu\textrm{g}\;ml^{-1}$) was 2 times higher than that of the $SH2-0.32-{\Delta}bcl-2$ culture (cells transfected with bcl-2-deficient plasmid, $10.5\;\mu\textrm{g}\;ml^{-1}$) in the presence of NaBu. To determine the effect of NaBu/Bcl-2 overexpression on the molecular integrity of protein products, antibodies purified from 14C6-bcl-2 and $SH2-0.32-{\Delta}bcl-2$ cultures in the presence of NaBu were characterized by using various molecular assay systems. For comparison, antibody purified from the parental rCHO cell culture (SH2-0.32) in the absence of NaBu was also characterized. No significant changes in molecular weight of antibodies could be observed by SDS-PAGE. From GlycoSep-N column analysis, it was found that the core oligosaccharide structure ($GlcNAc_2Man_3GlcNAc_2$) was not affected by NaBu/Bcl-2 overexpression, while the microheterogeneity of N-linked oligosaccharide structure was slightly affected. Compared with the antibody produced in the absence of NaBu, the proportion of neutral oligosaccharides was increased from 10% (14C6-bcl-2) to 16% ($SH2-0.32-{\Delta}bcl-2$) in the presence of NaBu, which was accompanied by the reduced proportion of acidic oligosaccharides, especially of monosialylated and disialylated forms. The changes in microheterogeneous oligoformal structures of antibody in turn affected the mobility of antibody isoforms in isoelectric focusing (IEF), resulting in the occurrence of some more basic antibody isoforms produced in the presence of NaBu. However, the antigen-antibody binding properties were not changed by alteration of glycosylation pattern. The competitive enzyme-linked immunosorbent assay (ELISA) showed that the antibody produced by NaBu/Bcl-2 overexpression maintained its antigen-antibody binding properties with binding affinity of about $2.5{\times}10^9{\;}M^{-1}$. Taken together, no significant effects of NaBu/Bcl-2 overexpression on the molecular integrity of antibodies, produced by using serum-free medium, could be observed by the molecular assay systems.

Mechanical and Oxygen Permeation Properties of Layered Double Hydroxide/Ethylene Vinyl Acetate Nanocomposite Membranes (Mg-Al Layered Double Hydroxide/Ethylene Vinyl Acetate 나노복합막의 기계적 특성과 기체투과 특성에 관한 연구)

  • Hwang, Ji-Young;Lee, Sang-Hyup;Lee, Jong-Suk;Hong, Se-Ryung;Lee, Hyun-Kyung
    • Membrane Journal
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    • v.23 no.2
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    • pp.151-158
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    • 2013
  • The effect of layered double hydroxides (LDH) on the gas separation properties of ethylene vinyl acetate copolymer was investigated. Mg-Al LDH/EVA nanocomposite membranes were prepared from solution intercalation using organically modified LDH (DS-LDH). Dodecyl sulfate (DS)-LDH was obtained by the intercalation of DS anion in the interlayer. The nanocomposite structure has been elucidated by X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM). XRD pattern clearly shows that the DS-LDH layers are disorderly well dispersed in the EVA matrix. The maximum tensile strength and elongation of the LDH/EVA nanocomposite membrane were found with the LDH content 3 wt%. The thermal properties of nanocompostie membrane were enhanced by the incorporation of LDH in EVA matrix. Gas permeation of LDH/EVA nanocomposite membranes with LDH contents of 1, 3, 5 wt% was studied for $O_2$ and $CO_2$ single gases. The presence of 3 wt% LDH decreased $O_2$ permeability by up to 53% compared to the EVA membrane. In spite of barrier property of nanocomposite membrane, however, the gas permeability for $CO_2$ was increased due to its strong affinity with the residual OH groups on the LDH.

Vegetation Structure of Lower Stratum and Pinus densiflora Natural Regeneration Features from Micro-topography Classification in Pinus densiflora Forest of Anmyeon-do Island (안면도 소나무림 내 미세지형구분을 통한 하층식생구조와 소나무 천연갱신 양상)

  • Byeon, Seong Yeob;Kim, Hyun Seop;Yun, Chung Weon
    • Journal of Korean Society of Forest Science
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    • v.108 no.2
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    • pp.189-199
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    • 2019
  • The forest management paradigm has recently shifted from focusing on commercial production to focusing on ecosystem management. Accordingly, a natural seedling regeneration method that has a naturally high affinity has attracted much attention in recent years. The aim of this study was to determine the relationship among various environmental factors, lower stratum vegetation, and seedling regeneration in Pinus densiflora forests. The survey site comprised 50 sectors divided using the line transect method, and the survey data were divided into those from wet habitat (19 sites) and dry habitat (31 sites), depending on the soil humidity, and were analyzed separately to show the close relationship between soil humidity and natural seedling regeneration. As a result, the dry habitat exhibited high seedling density (157,419 trees/ha), with the main species being Quercus serrata, Zanthoxylum piperitum, Smilax china, and Pueraria lobata, while wet habitat exhibited low seedling density (57,895 trees/ha), with the main species being Stephanandra incisa, Castanea crenata, Lespedeza maximowiczii, Lysimachia barystachys, Aralia elata, and Styrax japonicus. The height and root-collar diameter under wet conditions exhibited faster growth than those under dry conditions. Height growth by the root-collar diameter in dry habitat increased faster than that in wet habitat. It was also confirmed that seedling regeneration in wet habitat exhibited a rapid growth pattern 5 years after germination. These results suggest that the seedlings begin to grow more rapidly after a period of suppression by competition with surrounding plants. Considering an ecosystem or ecological management approach, specific practices, such as bush control and vine clearing in wet habitats, should be more intensively conducted, especially at the beginning of the management operations.

Zoning Permanent Basic Farmland Based on Artificial Immune System coupling with spatial constraints

  • Hua, Wang;Mengyu, Wang;Yuxin, Zhu;Jiqiang, Niu;Xueye, Chen;Yang, Zhang
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.15 no.5
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    • pp.1666-1689
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    • 2021
  • The red line of Permanent Basic Farmland is the most important part in the "three-line" demarcation of China's national territorial development plan. The scientific and reasonable delineation of the red line is a major strategic measure being taken by China to improve its ability to safeguard the practical interests of farmers and guarantee national food security. The delineation of Permanent Basic Farmland zoning (DPBFZ) is essentially a multi-objective optimization problem. However, the traditional method of demarcation does not take into account the synergistic development goals of conservation of cultivated land utilization, ecological conservation, or urban expansion. Therefore, this research introduces the idea of artificial immune optimization and proposes a multi-objective model of DPBFZ red line delineation based on a clone selection algorithm. This research proposes an objective functional system consisting of these three sub-objectives: optimal quality of cropland, spatially concentrated distribution, and stability of cropland. It also takes into consideration constraints such as the red line of ecological protection, topography, and space for major development projects. The mathematical formal expressions for the objectives and constraints are given in the paper, and a multi-objective optimal decision model with multiple constraints for the DPBFZ problem is constructed based on the clone selection algorithm. An antibody coding scheme was designed according to the spatial pattern of DPBFZ zoning. In addition, the antibody-antigen affinity function, the clone mechanism, and mutation strategy were constructed and improved to solve the DPBFZ problem with a spatial optimization feature. Finally, Tongxu County in Henan province was selected as the study area, and a controlled experiment was set up according to different target preferences. The results show that the model proposed in this paper is operational in the work of delineating DPBFZ. It not only avoids the adverse effects of subjective factors in the delineation process but also provides multiple scenarios DPBFZ layouts for decision makers by adjusting the weighting of the objective function.

Evaluation of Ecological Niche for Major Tree Species in the Natural Deciduous Forest of Mt. Chumbong (점봉산(點鳳山) 일대(一帶) 천연활엽수림(天然闊葉樹林)의 주요(主要) 구성(構成) 수종(樹種)에 대한 생태지위(生態地位) 평가(評價))

  • Kim, Guang Ze;Kim, Ji Hong
    • Journal of Korean Society of Forest Science
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    • v.90 no.3
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    • pp.380-387
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    • 2001
  • The characteristics of ecological niche, breadth and overlap, for seventeen major tree species were evaluated in the natural deciduous forest in Mt. Chumbong area. Employed by the plot sampling method, the environmental gradient for vertical niche was based on the intensity of light within the forest, and that for horizontal niche was based on multi-dimensional resources in distribution pattern. The result showed that Fraxinus rhynchophylla had the highest value of vertical niche breadth and Maackia amurensis had the lowest, and Acer pseudo-sieboldianum had the highest value of horizontal niche breadth and Betula costata had the lowest. There was no significant correlation between both measures of niche breadth. However, the tolerance index for each species was positively correlated to the values of niche breadth. Spearman's rank correlation coefficients were applied to test the correlationship between the species ranks of tolerance index and those of two ecological niche breadths. The coefficient of $r_s=0.432$ ($P{\leq}0.1$) was not enough to support significant correlationship between the tolerance index and vertical niche breadth at the 95% probability. If Carpinus cordata, rarely reach canopy of the forest due to its own growth form, are excluded from the analysis, coefficient was calculated as $r_s=0.650$ ($P{\leq}0.01$), resulting in highly significant correlationship. The Spearman's rank correlation coefficient was $r_s=0.797$ ($P{\leq}0.01$) for tolerance indices and the values of horizontal niche breadth, indicating highly significant. Four distinctive species groups, produced by cluster analysis on the basis of ecological niche overlap for each pair of species, were in considerable accord with the positively associated species constellation pattern created by the inter-species association analysis.

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The Selection of Appropriate Sampler for the Assessment of Macrobenthos Community in Saemangeum, the West Coast of Korea (새만금 외해역에서 대형 저서동물 군집 조사를 위한 적정 채집기의 선택)

  • 유재원;김창수;박미라;이형곤;이재학;홍재상
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.8 no.3
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    • pp.285-294
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    • 2003
  • To select an appropriate sampler for the environmental monitoring survey in coastal waters of Saemangeum, Jeollabuk-do, a macrobenthic sampling was conducted in April 2002. Employed samplers were dredge (type Charcot), a semi-quantitative sampler and Smith-McIntyre (SM) and van Veen grab (VV) as quantitative ones. One haul was tried for dredge and 3 replicates (0.1 ㎡${\times}$3) for SM and W at each of 11 stations. Comparisons of sediment volume in sampler bucket and of precision of biological parameters (i.e., density, biomass, species number and diversity index, H') were made between SM and VV. Sediment volume was significantly different (SM > VV) at p-value of 0.0050 (paired t-test) and, in average, 3 replicate samples of SM and VV satisfied a precision level of 0.2 by applying 4th root transformation. Patterns of observed and expected species numbers and H' were compared. Dredge-VV samples showed higher affinity than any other pair. Several dominant species in the area were underestimated in dredge samples (e.g., polychaete Heteromastus filiformis. Aricidea assimilis etc.). Quantifying the agreement pattern of multi-species responses was accomplished by estimating correlations between similarity matrices. Correlation between dredge and VV was slightly higher, but near-per-fect matches were found in general. Different ranks and composition among principal species lists were presumably linked to the effect of penetration depth that differs among samplers. Lower level of some species' abundance in VV samples (ca. 50% compared with those of SM) was explained in this context. It seem appropriate to regard the effect as a probable cause of relatively higher correlations in dredge-VV, Overall bio-logica1 features indicated that a better choice could be SM in situations of requiring high data quality. The others work well, however, on observing and defining faunal characteristics and their capability cannot be questionted if we do not expect a first-order quality.

Nutritional and Tissue Specificity of IGF-I and IGFBP-2 Gene Expression in Growing Chickens - A Review -

  • Kita, K.;Nagao, K.;Okumura, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.747-754
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    • 2005
  • Nutritional regulation of gene expression associated with growth and feeding behavior in avian species can become an important technique to improve poultry production according to the supply of nutrients in the diet. Insulin-like growth factor-I (IGF-I) found in chickens has been characterized to be a 70 amino acid polypeptide and plays an important role in growth and metabolism. Although it is been well known that IGF-I is highly associated with embryonic development and post-hatching growth, changes in the distribution of IGF-I gene expression throughout early- to late-embryogenesis have not been studied so far. We revealed that the developmental pattern of IGF-I gene expression during embryogenesis differed among various tissues. No bands of IGF-I mRNA were detected in embryonic liver at 7 days of incubation, and thereafter the amount of hepatic IGF-I mRNA was increased from 14 to 20 days of incubation. In eyes, a peak in IGF-I mRNA levels occurred at mid-embryogenesis, but by contrast, IGF-I mRNA was barely detectable in the heart throughout all incubation periods. In the muscle, no significant difference in IGF-I gene expression was observed during different stages of embryogenesis. After hatching, hepatic IGF-I gene expression as well as plasma IGF-I concentration increases rapidly with age, reaches a peak before sexual maturity, and then declines. The IGF-I gene expression is very sensitive to changes in nutritional conditions. Food-restriction and fasting decreased hepatic IGF-I gene expression and refeeding restored IGF-I gene expression to the level of fed chickens. Dietary protein is also a very strong factor in changing hepatic IGF-I gene expression. Refeeding with dietary protein alone successfully restored hepatic IGF-I gene expression of fasted chickens to the level of fed controls. In most circumstances, IGF-I makes a complex with specific high-affinity IGF-binding proteins (IGFBPs). So far, four different IGFBPs have been identified in avian species and the major IGFBP in chicken plasma has been reported to be IGFBP-2. We studied the relationship between nutritional status and IGFBP-2 gene expression in various tissues of young chickens. In the liver of fed chickens, almost no IGFBP-2 mRNA was detected. However, fasting markedly increased hepatic IGFBP-2 gene expression, and the level was reduced after refeeding. In the gizzard of well-fed young chickens, IGFBP-2 gene expression was detected and fasting significantly elevated gizzard IGFBP-2 mRNA levels to about double that of fed controls. After refeeding, gizzard IGFBP-2 gene expression decreased similar to hepatic IGFBP-2 gene expression. In the brain, IGFBP-2 mRNA was observed in fed chickens and had significantly decreased by fasting. In the kidney, IGFBP-2 gene expression was observed but not influenced by fasting and refeeding. Recently, we have demonstrated in vivo that gizzard and hepatic IGFBP-2 gene expression in fasted chickens was rapidly reduced by intravenous administration of insulin, as indicated that in young chickens the reduction in gizzard and hepatic IGFBP-2 gene expression in vivo stimulated by malnutrition may be, in part, regulated by means of the increase in plasma insulin concentration via an insulin-response element. The influence of dietary protein source (isolated soybean protein vs. casein) and the supplementation of essential amino acids on gizzard IGFBP-2 gene expression was examined. In both soybean protein and casein diet groups, the deficiency of essential amino acids stimulated chickens to increase gizzard IGFBP-2 gene expression. Although amino acid supplementation of a soybean protein diet significantly decreased gizzard IGFBP-2 mRNA levels, a similar reduction was not observed in chickens fed a casein diet supplemented with amino acids. This overview of nutritional regulation of IGF-I and IGFBP-2 gene expression in young chickens would serve for the establishment of the supply of nutrients to diets to improve poultry production.

Vertical Distribution of Vascular Plant Species along an Elevational Gradients in the Gyebangsan Area of Odaesan National Park (오대산국립공원 계방산지구 관속식물의 고도별 수직분포)

  • An, Ji-Hong;Park, Hwan-Jun;Nam, Gi-Heum;Lee, Byoung-Yoon;Park, Chan-Ho;Kim, Jung-Hyun
    • Korean Journal of Ecology and Environment
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    • v.50 no.4
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    • pp.381-402
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    • 2017
  • In order to investigate distribution of vascular plants along elevational gradient in the Nodong valley of Gyebangsan, vascular plants of eight sections with 100-meter-high were surveyed from the Auto-camping site (800 m) to the top of a mountain (1,577 m). There were a total of 382 taxa: 89 families, 234 genera, 339 species, 7 subspecies, 34 varieties, and 2 forms. As a result of analyzing the pattern of species richness, it showed a reversed hump-shaped with minimum richness at mid-high elevation. As a result of analyzing habitat affinity types, the proportion of forest species increased with increasing elevation. But, the ruderal species decreased with increasing elevation, and then increased at the top of a mountain. As for the proportion of life forms, the annual herbs gradually decreased with increasing elevation, but it did not appear between 1,300 m and 1,500 m and then increased at the top of a mountain. The trees gradually increased with elevation and decreased from 1,300~1,400 m. The vascular plants divided into four groups by using DCA. The arrangement of each stands was arranged in order from right to left on the I axis according to the elevation. The distribution of vascular plants is determined by their own optimal ranges of vegetation. Also, rise in temperature due to climate change affects the distribution of vascular plants, composition, and diversity. Therefore, continuous monitoring is necessary to confirm ecological and environmental characteristics of vegetation, distribution ranges, changes of habitat. Furthermore, plans for conservation and management based on these data should be prepared according to climate change.