• Title/Summary/Keyword: tissue residues

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Lectin histochemistry of lung tissues in the Streptozotocin rat fetus (Streptozotocin을 투여한 흰쥐 태자 폐조직의 렉틴 조직화학 염색성)

  • Hong, Hea-Nam;Kim, Dong-Hou
    • Applied Microscopy
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    • v.23 no.2
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    • pp.84-96
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    • 1993
  • This study was to investigate the effects of maternal diabetes on the lung tissue of the fetal rat using lectin histochemistry and electron microscope technique. Maternal diabetes was induced by intraperitoneal injection of streptozotocin (75 mg/kg the body weight) into pregnant Sprague-Dawley rats on the 7th day of gestation. Fetuses of streptozotocin induced diabetic rats exhibited delayed lung maturation and reduced air space. In lectin histochemistry, the binding of Maclura pomifera (MPA) to fetal lungs from diabetic mothers was reduced, but no significant changes in the bindings of Concanavalin A (Con A), Wheat germ agglutinin (WGA), Ricinus communis I (RCA I) and Griffonia simplicifolia (GSI-$B_4$) were noted. Because the MPA has affinity to terminal N-acetyl-D-galactosamine residues constantly linked O-glycosidically to serine or threonine, the present findings may indicates that maternal diabetes interfere with the processes of O-linked glycosylation in fetal rat lung.

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Backbone assignment of human Hoxc9DBD

  • Ja-Shil Hyun;Sung Jean Park
    • Journal of the Korean Magnetic Resonance Society
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    • v.27 no.4
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    • pp.23-27
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    • 2023
  • Hoxc, or the Homeobox C cluster, is a group of genes that play a crucial role in embryonic development, particularly in patterning the body along the anterior-posterior axis. These genes encode transcription factors, which are proteins that bind to DNA and regulate the expression of other genes. Hoxc9 is specifically involved in the development of the skeletal system, nervous system, and adipose tissue. Hoxc9 overexpression has been linked to the development of various cancers such as leukemia and breast cancer. Here, we assigned the chemical shifts Hoxc9 DNA binding domain (DBD) using heteronuclear NMR techniques. The helical regions of Hoxc9 DBD correspond to the residues T200 - F213 (Helix I), T218 - L229 (Helix II), and T232 - K249 (Helix III). Our result would be helpful for studing the molecular interactions of the Hoxc9 DBD and other proteins.

Comparison between the liquid-liquid partition method and modified QuEChERS method for the analysis of pesticide residues in beef fat (소지방 중 잔류농약분석을 위한 액-액분배법과 modified QuEChERS법 비교)

  • Kim, Yoen-Joo;Choi, Yoon-Hwa;Shin, Bang-Woo;Lee, Jung-Hark
    • Korean Journal of Veterinary Service
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    • v.34 no.4
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    • pp.429-439
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    • 2011
  • This article described the comparison of a quick, easy, cheap, effective, rugged and safe (QuEChERS) sample preparation and the classical method established by National Veterinary Research and Quarantine Service (NVRQS) for the determination of pesticide residues in livestock products using GC-tandem mass spectrometry. The classical method by NVRQS used liquid-liquid partioning followed by evaporizing. The modified QuEChERS entailed extraction of 2 g sample with 15 ml acetonitrile containing 1% acetic acid followed by addition of 6 g anhydrous magnesium sulfate and 1.5 g sodium acetate. After centrifugation, 6 ml of the extract underwent a cleanup step (in a technique known as column-based solid phase extraction) using 400 mg each of $C_{18}$ and primary secondary amine sorbents plus 1,200 mg magnesium sulfate. The quantitation of individual pesticides by both methods was based on tissue standard calibration curves with a correlation coefficient in excess of 0.98 for the 24 pesticides. The detection limits by the classical method were ranged 1.3~5.0 ${\mu}g$/kg, with mean recoveries between 76.2% and 114.3% except aldrin (59.3%) and deltamethrin (63.6%). The detection limits by modified QuEChERS were ranged 0.3~6.2 ${\mu}g$/kg, with mean recoveries between 68.0% and 114.3% except dimethipin (152.6%), chlorfenvinphos (138.1%), 4,4-DDT (61.5%), aldrin (60.4%) and chinomethionate (30.3%).

Characterization of Expressed Sequence Tags (ESTs) Generated from the Bombyx mandarina Whole Larvae and Molecular Cloning of Serine Protease Homologue Gene

  • Hwang, Jae Sam;Yun, Eun Young;Goo, Tae Won;Kim, Iksoo;Choi, Kwang Ho;Seong, Su Il;Kim, Keun Young;Lee, Sang Mong;Kang, Seok Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.2
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    • pp.167-171
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    • 2004
  • We constructed an oligo-d(T) primed directional cDNA library from the Bombyx mandarina whole larvae. In an effort to isolate genes expressed in the B. mandarina, 227 expressed sequence tags (ESTs) were generated by single-pass sequencing from the cDNA library. Sequence analysis showed that 107 clones (47.1%) were classified into known genes and 120 clones (52.9%) were novel transcripts, which are unknown for their function. Of the 107 known genes, the most abundant gene was found to be actin and followed by serine protease in the expression profile. Among these clones, a serine protease homolog (BmSP) which is a class of proteolytic enzymes isolated. Full-length sequence of the BmSP cDNA clone was 922 bp in length and has an open reading frame of 276 amino acids. The conserved histidine, aspatic acid and serine residues forming the catalytic center as well as cysteine residues contributing to three disulphide bonds also were found in Bmsp gene. mRNA expression analysis revealed a high and specific expression of the gene only in midgut tissue, suggesting that BmSP gene is closely associated with the expression of digestive enzyme.

Persistence and Distribution of Trunk-Injected Abamectin in Pinus thunbergii and Pinus koraiensis Tissues (수간주입한 아바멕틴의 곰솔과 잣나무 내 분포와 지속성)

  • Lee, Sang-Myeong;Kim, Dong-Soo;Kim, Chul-Su;Cho, Kyu-Seong;Choo, Ho-Yul;Lee, Dong-Woon
    • The Korean Journal of Pesticide Science
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    • v.13 no.3
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    • pp.190-196
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    • 2009
  • The residues of abamectin 1.8% EC, resisted for control of pine wood nematode, Bursaphelenchus xylophilus in pine tree were surveyed in tissue of Pinus thunbergii and P. koraiensis after injection of a liquid formulation. Limits of detection of abamectin in tissue of P. thunbergii were $0.05\;mg\;kg^{-1}$ and mean recoveries at $0.5\;mg\;kg^{-1}$ trunk injection were 90.9% and 93.1% respectively in stem and trunk of P. thunbergii. Abamectin 1.8% EC, trunk injected in 15 m height P. thunbergii were detected in all stem (edible part of carrier insect of pine wood nematode, Monochamus alternatus) from 0.29 to $0.73\;mg\;kg^{-1}$ after 150 days injection. Amount of residue of abamectin 1.8% EC in 12.6 cm mean breast height diameter (DBH) P. thunbergii were variable depending on individual trees in natural forest. Amount of residues in lower and middle part of trunk were reduced with the passage of the injection time. In upper part of trunk were detected $1.84\;mg\;kg^{-1}$ on 30 days after injection however $0.65\;mg\;kg^{-1}$ on 15 days after injection and under detection limit on 100 and 180 days after injection in P. thunbergii. Bottom and middle parts of crown were detected $0.183\;mg\;kg^{-1}$ and $0.173\;mg\;kg^{-1}$ respectively on 180 days after injection in P. thunbergii. Mean residues of abamectin in crown and trunk were $0.80\;mg\;kg^{-1}$ and $0.30\;mg\;kg^{-1}$ on 170 days after trunk injection in 20 cm DBH and 9 m height P. koraiensis. Mean residues of abamectin in crown and trunk were $0.67\;mg\;kg^{-1}$ and $0.36\;mg\;kg^{-1}$ on 170 days after trunk injection in 15 cm DBH and 6 m height P. koraiensis.

Anti-calcification of Bovine Pericardium for Bioprosthetic Heart Valves after Surface Modification with Hyaluronic Acid Derivatives

  • Hahn Sei Kwang;Ohri Rachit;Giachelli Cecilia M.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.218-224
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    • 2005
  • Surface modification of glutaraldehyde fixed bovine pericardium (GFBP) was success­fully carried out with hyaluronic acid (HA) derivatives. At first, HA was chemically modified with adipic dihydrazide (ADH) to introduce hydrazide functional group into the carboxyl group of HA backbone. Then, GFBP was surface modified by grafting HA-ADH to the free aldehyde groups on the tissue and the subsequent HA-ADH hydrogel coating. HA-ADH hydrogels could be prepared through selective crosslinking at low pH between hydrazide groups of HA-ADH and crosslinkers containing succinimmidyl moieties with minimized protein denaturation. When HA­ADH hydrogels were prepared at low pH of 4.8 in the presence of erythropoietin (EPO) as a model protein, EPO release was continued up to $85\%$ of total amount of loaded EPO for 4 days. To the contrary, only $30\%$ of EPO was released from HA-ADH hydrogels prepared at pH=7.4, which might be due to the denaturation of EPO during the crosslinking reaction. Because the carboxyl groups on the glucuronic acid residues are recognition sites for HA degradation by hyaluronidase, the HA-ADH hydrogels degraded more slowly than HA hydrogels prepared by the crosslinking reaction of divinyl sulfone with hydroxyl groups of HA. Following a two-week subcutaneous implantation in osteopontin-null mice, clinically significant levels of calcification were observed for the positive controls without any surface modification. However, the calcification of surface modified GFBP with HA-ADH and HA-ADH hydrogels was drastically reduced by more than $85\%$ of the positive controls. The anti-calcification effect of HA surface modification was also confirmed by microscopic analysis of explanted tissue after staining with Alizarin Red S for calcium, which followed the trend as observed with calcium quantification.

Isolation and characterization of Brcpi1 gene encoding phytocystatin from chinese cabbage (Brassica rapa L.) seedlings (배추 유래 phytocystatin 유전자, Brcpi1의 분리 및 발현특성 분석)

  • Jung, Yu-Jin;Cho, Yong-Gu;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.407-414
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    • 2009
  • A cDNA clone encoding phytocystatin was isolated from Brassica rapa seedlings, through rapid amplification of cDNA ends (RACE). This gene (name as Brcpi1; GenBank accession no.: EF079953) had a total length of 881 bp with an open reading frame of 609 bp, and encoded predicted polypeptide of 203 amino acid (aa) residues including a putative N-terminal signal peptide. Other relevant regions found its sequence included the G and PW conserved aa motifs, and the consensus LARFAV sequence for phytocystatins and the reactive site QVVAG. The BrCPI1 protein shared 95, 94, 81, 80 and 78% identity with other CPI proterins isolated from Brassica oleracea (BoCPI-1), Arabidopsis thaliana (AtCY SB), Glycine max (GmCPI), Oryza sativa (OsCYS-2) and Zea may (ZmCPI) at amino acid level, respectively. Southern blot analysis showed that Brcpi1 was a low copy gene. Expression pattern analysis revealed that Brcpi1 was a tissue-specific expressing gene during reproductive growth and strongly expressed at mature seedling stages. Furthermore, overexpression of Brcpi1 in transgenic Arabidopsis was enhanced tolerance to salt and cold stresses. Meanwhile the juvenile seedling of Brcpi1 transgenic plants was not affected by various concentrations ABA in MS medium. Taken together, the results showed that Brcpi1 functioned as a cysteine protease inhibitor and it exhibited a protective agent against diverse types of abiotic stress, which induced this gene in a tissue- and stress-specific manner.

Mutation of Angiogenesis Inhibitor TK1-2 to Avoid Antigenicity In Vivo

  • Lee Sang-Bae;Kim Hyun-Kyung;Oh Ho-Kyun;Hong Yong-Kil;Joe Young-Ae
    • Biomolecules & Therapeutics
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    • v.14 no.1
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    • pp.30-35
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    • 2006
  • Tissue-type plasminogen activator (t-PA) is a multidomain serine protease containing two kringle domains, TK1-2. Previously, Pichia-derived recombinant human TK1-2 has been reported as an angiogenesis inhibitor although t-PA plays an important role in endothelial and tumor cell invasion. In this work, in order to improve in vivo efficacy of TK1-2 through elimination of immune reactivity, we mutated wild type TK1-2 into non-glycosylated form (NE-TK1-2) and examined whether it retains anti-angiogenic activity. The plasmid expressing NE-TK1-2 was constructed by replacing $Asn^{l17}\;and\;Asn^{184}$ with glutamic acid residues. After expression in Pichia pastoris, the secreted protein was purified from the culture broth using S-sepharose and UNO S1-FPLC column. The mass spectrum of NE-TK1-2 showed closely neighboring two peaks, 19631.87 and 19,835.44 Da, and it migrated as one band in SDS-PAGE. The patterns of CD-spectra of these two proteins were almost identical. Functionally, purified NE-TK1-2 was shown to inhibit endothelial cell migration in response to bFGF stimulation at the almost same level as wild type TK1-2. Therefore, the results suggest that non-glycosylated NETK1-2 can be developed as an effective anti-angiogenic and anti-tumor agent devoid of immune reactivity.

Characterization and Expression Pattern of the Partial Myostatin cDNA in Shrimp, Fenneropenaeus chinensis

  • Lee, Sang Beum;Kim, Yong Soo;Yoon, Moongeun;Kim, Su-Kyoung;Jang, In Kwon;Lim, Hyun Jeong;Jin, Hyung-Joo
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.4
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    • pp.224-229
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    • 2007
  • Muscle tissue expresses many muscle-specific genes, including myostatin (also known as GDF8) that is a member of the transforming growth factor-beta superfamily. Myostatin (MSTN) negatively regulates mammalian skeletal muscle growth and development by inhibiting myoblast proliferation. Mice and cattle possessing mutant MSTN alleles display a 'double muscling' phenotype characterized by extreme skeletal muscle hypertrophy and/or hyperplasia. In this study, we first have characterized partial cDNA of a MSTN gene from the muscle tissue in the F. chinensis and examined its expression pattern in various tissues. The partial MSTN gene (GenBank accession number EU 131093) in the F. chinensis was 1134 bp, encoding for 377 amino acids that showed 63-93% amino acid similarity to other vertebrate MSTNs, containing a conserved proteolytic cleavage site (RXRR) and conserved cysteine residues in the C-terminus. Based on a RT-PCR, the MSTN gene was expressed in the all tissues of F. chinensis used in this study.

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Study on the changes of sulfamethnzine residues in serum and practical organs of rats orally administrated with sulfamethnzine sodium (Rat에 sulfamethazine sodium 경구투여 후 혈청 및 실질장기내 sulfamethazine의 잔류량 추이에 관한 연구)

  • 도재철;이영미;조민희;신상희;박희주;송희종;정종식
    • Korean Journal of Veterinary Service
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    • v.23 no.4
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    • pp.321-333
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    • 2000
  • In order to know the depletive changes of sulfamethazine residues in senlm and practical organs of rats orally administered with sulfamethazine sodium(SMS), the concentration of sulfamethazine was measured in serum and tissue(kidney, liver, spleen, testis, and skeletal muscle) of rats with using high performance liquid chromatography(HPLC). SMS was orally administrated to sprague-dawley male rats(body weight, 200~300g) with using sonde at the rate of 20mg/100g body weight(recommended therapeutic dose) on once a day for 3 days. There were investigated the depletive changes of the sulfamethazine in serum, kidney, liver, spleen, testis and skeletal muscle of rat at the time 8 hours, 1st, 2nd, 3rd, 4th, 5th and 6th day after administration SMS, respectively. The results obtained were summarized as follows; 1. After oral administration of the SMS, the mean concentrations of sulfamethazine in serum according to the time lapsed were showed 215.53$\pm$42.99ppm at the 8 hours after withdrawal of medicated sulfamethazine. And gradually according to the time lapsed, the concentrations of sulfamethazine residues in serum were significantly (p<.05) decreased 25.87$\pm$5.18ppm at 1st day, 2.30$\pm$0.61ppm at 3rd day and 0.11$\pm$0.02ppm at 6th day respectively. 2. The mean concentrations of sulfamethazine in kidney, liver, spleen, muscle and testis according to the time lapsed after administration SMS were showed 83.82$\pm$12.16, 81.77$\pm$12.88, 36.96$\pm$5.35, 35.96$\pm$TEX>$\pm$1.39 and 27.89$\pm$1.92 ppm at the 8 hours, respectively. And gradually according to the time lapsed, the concentrations of sulfamethazine residues in the each of samples were significantly(p<.05) decreased such as 7.15$\pm$0.26, 5.62$\pm$0.72, 2.43$\pm$0.29, 1.99$\pm$0.14 and 3.11$\pm$0.48 ppm at 1st day, 0.52$\pm$0.04, 1.32$\pm$0.22, 0.13$\pm$0.03, 0.15$\pm$0.06 and 0.26$\pm$0.11ppm at 3rd day, and 0.03$\pm$0.01, 0.11$\pm$0.03, 0.02$\pm$0.01, 0.009$\pm$0.001 and 0.02$\pm$0.01 ppm at 6th day, respectively. 3. After oral administration of the SMS to rats, the residual concentrations of sulfamethazine in skeletal muscle were significantly (p<.05) decreased 35.96$\pm$1.39 to 0.009$\pm$$\pm$0.001 ppm between 8 hours and 6th day, respectively From the 4th day, the residual concentrations of sulfamethazine were showed 0.10$\pm$0.04 ppm below 0.1 ppm at the permitted limit concentration of muscle in Korea. In conclusion, this study could be suggested the relationship between administrated period, doses of sulfonamides and residual aspects of serum and practical organs, and the importance of observing ceasing period of antibiotic drugs before forwarding livestocks to slaughter.

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