• Title/Summary/Keyword: cell alignment

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PCR-SSCP of Serum Lysozyme Gene (Exon-III) in Riverine Buffalo and Its Association with Lysozyme Activity and Somatic Cell Count

  • Sahoo, Nihar Ranjan;Kumar, Pushpendra;Bhushan, Bharat;Bhattacharya, T.K.;Sharma, Arjava;Dayal, Sanker;Pankaj, Prabhat Kumar;Sahoo, Monalisa
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.8
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    • pp.993-999
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    • 2010
  • Serum lysozyme gene is one of the important genes influencing the immune system as its product can cause lysis of bacterial cell wall by cleaving the peptidoglycan layer. The present investigation on the serum lysozyme gene of Indian riverine buffalo was undertaken with the objectives to identify and characterize single nucleotide polymorphic patterns by PCR-SSCP method as well as to study the effect of different genotypes on serum lysozyme activity and somatic cell count. A total of 280 animals comprising four different famous bubaline breeds (Murrah, Mehsana, Surti and Bhadawari), spread over six different farms across the country were used for this study. A 276 bp (partial intron 2, complete exon 3 and partial intron 3) fragment of lysozyme gene was screened for polymorphism using the SSCP technique. Four genotypes namely AA, AB, BC and AC were observed, out of which BC genotype was found to be the most frequent. Among these three alleles, C allele (0.38) was most prevalent in these populations. Various SSCP allelic variants were cloned for sequencing and sequences were submitted to NCBI Genbank. From the alignment of the nucleotide sequences of various allelic variants, it was found that there were differences in 12 positions among the alleles, out of which maximum variation (at 8 places) was found in the intronic region. The allele A was closer to allele-C than allele-B. Allele B was phylogenetically equidistant from both of the other alleles. Mean lysozyme activity determined in serum samples of different animals of Murrah buffalo was $27.35{\pm}2.42\;{\mu}g$ per ml of serum, whereas the mean somatic cell count was $1.25{\pm}0.13{\times}10^5$ cells per ml of milk. The SSCP pattern-wise effects of various genotypes on lysozyme activity and SCC were analyzed. Although the mean values were apparently different in various genotypes, these differences were statistically non-significant. It can be concluded that the riverine buffaloes are sufficiently polymorphic with respect to serum lysozyme gene. The absence of AA genotype in Bhadawari breed of buffalo can be considered as a marker for breed characterization. The difference of four nucleotides in exon-3 indicates high selection pressure on the gene.

Electro-optic characteristics of novel biased vertical alignment device using the polymerized reactive mesogen (광경화성 단분자를 이용한 새로운 수직배향 액정 디바이스의 전기 광학적 특성연구)

  • Kim, Dae-Hyun;Kim, Sung-Min;Cho, In-Young;Kim, Woo-Il;Kwon, Dong-Won;Son, Jong-Ho;Ryu, Jae-Jin;Kim, Kyeong-Hyeon;Lee, Seung-Hee
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2009.06a
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    • pp.269-270
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    • 2009
  • The biased vertical alignment (BVA) liquid crystal (LC) mode shows a has a distinct advantage of lower manufacture cost due to the elimination of a lithographic process step to form either ITO-patterning or protrusions on the color-filter substrates. However, those devices have complex voltage conditions which is the respective induce voltage on common electrode, pixel electrode and bias electrode when positive and negative frame. In order to overcome the complex voltage condition, the pretilt angles is controlled by photo polymerization of the UV-curable reactive mesogen (RM). According to our studies, voltages to the cell are critical to achieve an optimized surface-modified quality BVA (Q-BVA) mode which provides the well defined reorientation of the LCs with respect to an electric field.

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Healing Effects of Astragali Radix Extracts on Experimental Open Wounds in Rats (흰쥐의 외과적 창상에 대한 황기(黃耆) 추출액의 치료효과)

  • Han, Dong-Oh;Kim, Gun-Ho;Choi, Yong-Bok;Shim, In-Sop;Lee, Hye-Jung;Lee, Yong-Gheun;Kim, Jang-Hyun;Chang, Gyu-Tae;Hahm, Dae-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.1
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    • pp.92-97
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    • 2005
  • Inhibition of inflammatory response, acceleration of basal cell growth, and balanced synthesis of extracellular matrix (ECM) are important in healing of cutaneous open wounds. In order to evaluate the healing effects of water extracts of Radix Astragali (the root of Astragalus membranaceus (Fisch.)) on open wound at early stage, the experimental open wounds were generated on the dorsal sides of SD rats under anesthesia. The boiled-water extracts of Radix Astragali $(100{\mu}l)$, soaked into an occlusive film dressing were applied once a day for eleven consecutive days. The healing process was assessed by measuring macroscopic appearance and wound areas of the open wounds. The molecular aspects of healing process by Radix Astragali extracts were also investigated by Hematoxylin-Eosin (H-E) double staining and immunohistological staining of collagen type I in the healed skin area, implying cell density and linear alignment of the granulation tissue, and ECM synthesis and its remodeling, respectively. The Astragali radix extracts were found to significantly accelerate the cutaneous wound healing by suppressing the inflammation and stimulating the basal cell growth in wounded area, as compared to epidermal growth factor (EGF).

Study on Electro-optic Characteristics of the Optically Compensated Bend Liquid Crystal Display Using UV Curable Monomer (광경화성 단분자를 이용한 광학 보상 휨 액정 디스플레이의 전기광학 특성연구)

  • Lim, Young-Jin;Jeon, Eun-Jeong;Kwon, Dong-Won;Kim, Jeong-Hwan;Jeong, Kwang-Un;Lee, Myong-Hoon;Lee, Seung-Hee
    • Polymer(Korea)
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    • v.33 no.5
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    • pp.496-500
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    • 2009
  • Optically compensated bend liquid crystal display (OCB-LCD) has many application fields owing to its fast response time and wide viewing angle. However, in order to operate the OCB-LCD in bend state, this device needs quick transitions from the initial splay state to bend state. Unlike conventional approach using transient high voltage for the transition, the OCB-LCD with high surface tilt angle, which was achieved by polymerization of UV curable reactive mesogen monomer under certain voltage, was manufactured and the cell showed bend state initially. Electro-optic and electrical characteristics of the cell were analyzed. The cell shows a fast response time owing to high surface pretilt angle and very low residual DC less than 0.1 V although another polymer layer is formed above polymer alignment layers.

Evaluation of Consolidation Properties in Soft Soils Using Elastic and Electromagnetic Waves (전단파와 전자기파를 이용한 연약 지반의 실내 압밀 특성 평가)

  • Lee, Chang-Ho;Yoon, Hyung-Koo;Kim, Joon-Han;Lee, Jong-Sub
    • Journal of the Korean Geotechnical Society
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    • v.24 no.8
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    • pp.25-34
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    • 2008
  • A new hybrid oedometer cell is designed and manufactured to investigate a behavior of soft soils by using elastic and electromagnetic waves during consolidation test. Bender elements, which generate and detect shear waves, are placed in the top cap and the bottom plate and mounted on the oedometer wall. Double wedge type electrical resistance probe, which measures local void ratio change, is positioned onto the top cap of the oedometer cell. The bender elements and the electrical resistance probe are anchored into a nylon set screw with epoxy resin. The nylon set screw with epoxy resin minimizes directly transmited elastic waves through the oedometer cell due to impedence mismatch and allows for easy replacement of defected bender elements and electrical resistance probe. Primary consolidation time can be estimated from the slope of electrical resistance versus log time curve and the evolution of shear wave velocity. The shear wave velocity can be used to assess inherent anisotropy when disturbance effects are minimized because particle alignment affects the shear wave velocity. The void ratios evaluated by the electrical resistance probe are similar to those by the settlement during consolidation. This study suggests that the shear wave velocity and the electrical resistance can provide complementary imformations to understand consolidation characteristics such as primary consolidation, anisotropy, and void ratio.

The Expression Analysis of Complement Component C3 during Early Developmental Stages in Olive Flounder (Paralichthys olivaceus)

  • Lee, Jang-Wook;Lee, Young Mee;Lee, Jeong-Ho;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Hwang, In Joon;Kim, Sung Yeon
    • Development and Reproduction
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    • v.17 no.4
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    • pp.311-319
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    • 2013
  • Fish larvae are immediately exposed to microbes from hatching to maturation of their lymphoid organs, therefore effective innate mechanisms is very important for survival in such an environment. The key component of innate immune system, C3 is central protein of all activation pathways of the complement system, leading to inflammatory reactions, such as opsonisation, chemotaxis, and cell lysis of pathogens. Although, innate mechanisms is essential for survival in the early stage of development, little is known about defence mechanisms. In this study, the alignment analysis showed that amino acid sequence of C3 from olive flounder liver EST homologous to other known C3 sequences with 73-99% identity. Also, we examined the tissue distribution of olive flounder C3 and analyzed expression pattern from the fertilized egg until 28 days post hatching. As a result, olive flounder C3 mRNA was expressed only in the liver and the mRNA level more increased as developmental proceed during the early stage. These results may suggest that olive flounder C3 plays an important function in the early immune response of olive flounder larvae.

Stage and Tissue Specific Expression of Four TCR Subunits in Olive Flounder (Paralichthys olivaceus)

  • Lee, Young Mee;Lee, Jeong-Ho;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Hwang, In Joon;Kim, Sung Yeon
    • Development and Reproduction
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    • v.17 no.4
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    • pp.329-335
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    • 2013
  • TCR subunits are members of membrane-bound receptors which allow the fast and efficient elimination of the specific fish pathogens have regulated function in adaptive immunity. Sequence structure of TCR subunits have been reported for various teleosts, but the information of each TCR subunit functional characterization through expression analysis in fish was unknown. In this study, we examined the gene expression of TCR subunits in the early developmental stages and observed transcript levels in various tissues from healthy adult olive flounder by RT-PCR. The mRNA expression of alpha subunit was already detected in the previous hatching step. But the transcripts of another TCR subunit were not observed during embryo development and increased after hatching and maintained until metamorphosis at the same level. It was found that all TCR subunits mRNAs are commonly expressed in the immune-related organ such as spleen, kidney and gill, also weak expressed in fin and eye. TCR alpha and beta subunit were expressed in brain, whereas gamma and delta were not expressed same tissue. The sequence alignment analysis shows that there are more than 80% sequence homology between TCR subunits. Because it has a high similarity of amino acid sequence to expect similar in function, but expression analysis show that will have may functional diversity due to different time and place of expression.

Fabrication of Micro Patterned Fibronectin for Studying Adhesion and Alignment Behavior of Human Dermal Fibroblasts

  • Lee, Seung-Jae;Son, Young-Sook;Kim, Chun-Ho;Choi, Man-Soo
    • Macromolecular Research
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    • v.15 no.4
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    • pp.348-356
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    • 2007
  • The aim of this study was to fabricate a submicro-and micro-patterned fibronectin coated wafer for a cell culture, which allows the positions and dimensions of the attached cells to be controlled. A replica molding was made into silicon via a photomask in quartz, using E-beam lithography, and then fabricated a polydimethylsiloxane stamp using the designed silicon mold. Hexadecanethiol $[HS(CH_2){_{15}}CH_3]$, adsorbed on the raised plateau of the surface of polydimethylsiloxane stamp, was contact-printed to form self-assembled monolayers (SAMs) of hexadecanethiolate on the surface of an Au-coated glass wafer. In order to form another SAM for control of the surface wafer properties, a hydrophilic hexa (ethylene glycol) terminated alkanethiol $[HS(CH_2){_{11}}(OCH_2CH_2){_6}OH]$ was also synthesized. The structural changes were confirmed using UV and $^1H-NMR$ spectroscopies. A SAM terminated in the hexa(ethylene glycol) groups was subsequently formed on the bare gold remaining on the surface of the Aucoated glass wafer. In order to aid the attachment of cells, fibronectin was adsorbed onto the resulting wafer, with the pattern formed on the gold-coated wafer confirmed using immunofluorescence staining against fibronectin. Fibronectin was adsorbed only onto the SAMs terminated in the methyl groups of the substrate. The hexa (ethylene glycol)-terminated regions resisted the adsorption of protein. Human dermal fibroblasts (P=4), obtained from newborn foreskin, only attached to the fibronectin-coated, methyl-terminated hydrophobic regions of the patterned SAMs. N-HDFs were more actively adhered, and spread in a pattern spacing below $14{\mu}m$, rather than above $17{\mu}m$, could easily migrate on the substrate containing spacing of $10{\mu}m$ or less between the strip lines.

Molecular cloning and expression analysis of annexin A2 gene in sika deer antler tip

  • Xia, Yanling;Qu, Haomiao;Lu, Binshan;Zhang, Qiang;Li, Heping
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.4
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    • pp.467-472
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    • 2018
  • Objective: Molecular cloning and bioinformatics analysis of annexin A2 (ANXA2) gene in sika deer antler tip were conducted. The role of ANXA2 gene in the growth and development of the antler were analyzed initially. Methods: The reverse transcriptase polymerase chain reaction (RT-PCR) was used to clone the cDNA sequence of the ANXA2 gene from antler tip of sika deer (Cervus Nippon hortulorum) and the bioinformatics methods were applied to analyze the amino acid sequence of Anxa2 protein. The mRNA expression levels of the ANXA2 gene in different growth stages were examined by real time reverse transcriptase polymerase chain reaction (real time RT-PCR). Results: The nucleotide sequence analysis revealed an open reading frame of 1,020 bp encoding 339 amino acids long protein of calculated molecular weight 38.6 kDa and isoelectric point 6.09. Homologous sequence alignment and phylogenetic analysis indicated that the Anxa2 mature protein of sika deer had the closest genetic distance with Cervus elaphus and Bos mutus. Real time RT-PCR results showed that the gene had differential expression levels in different growth stages, and the expression level of the ANXA2 gene was the highest at metaphase (rapid growing period). Conclusion: ANXA2 gene may promote the cell proliferation, and the finding suggested Anxa2 as an important candidate for regulating the growth and development of deer antler.

Cloning and Characterization of the HSP70 Gene, and Its Expression in Response to Diapauses and Thermal Stress in the Onion Maggot, Delia antiqua

  • Chen, Bin;Kayukawa, Takumi;Monteiro, Antonia;Ishikawa, Yukio
    • BMB Reports
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    • v.39 no.6
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    • pp.749-758
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    • 2006
  • The cytosolic members of the HSP70 family of proteins play key roles in the molecular chaperone machinery of the cell. In the study we cloned and sequenced the full-length cDNA of Delia antiqua HSP70 gene, which is 2461 bp long and encodes 643 a.a. with a calculated molecular mass of 70,787 Da. We investigated gene copies of cytosolic HSP70 members of 4 insect species with complete genome available, and found that they are quite variable with species. In order to characterize this protein we carried out an alignment and a phylogenetic analysis with 41 complete protein sequences from insects. The analysis divided the cytosolic members of the family into two classes, HSP70 and HSC70, distinguishable on the basis of 15 residues. HSP70 class members were slightly shorter in length and smaller in molecular mass relative to the HSC70 class members, and the conservative and functional regions in these sequences were documented. Mainly, we investigated the expression of Delia antiqua HSP70 gene, in response to diapauses and thermal stresses. Both summer and winter diapauses elevated HSP70 transcript levels. Cold-stress led to increased HSP70 expression levels in summer- and winter-diapausing pupae, but heat-stress elevated the levels only in the winter-diapausing pupae. In all cases, the expression levels, after being elevated, gradually decreased with time. HSP70 expression was low in non-diapausing pupae but was up-regulated following cold- and heat-stresses. Heat-stress gradually increased the mRNA level with time whereas cold-stress gradually decreased levels after an initial increase.