• Title/Summary/Keyword: wheat germ agglutinin

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Preparation and Characterization of Wheat Germ Agglutinin-conjugated Praecoxin A (Wheat Germ Agglutinin-conjugated Praecoxin A의 제작 및 특성)

  • 김완수;김만석;김범수;이민원;이도익
    • YAKHAK HOEJI
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    • v.45 no.3
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    • pp.302-309
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    • 2001
  • Wheat germ agglutinin (WGA) pectin, which binds to human melanoma cell line, was conjugated with Praecoxin A using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide as a cross-linking agent. Physical mixture (PM) of WGA and Praecoxin A was also prepared by using a non-specific binding property of Praecoxin A to WGA. The WGA:Praecoxin A ratio in the conjugate and PM was approximately 1:18 and 1:20, respectively. The results of hemagglutination assay and enzyme-linked lectin assay indicated that the conjugate and PM maintained the lectin-like properties of the WGA. The binding ratio of conjugate was about 70% during 4-24 hr, but the most of Praecoxin A was released within 24 hr in the case of PM. These results lead to the conclusion that the conjugate is potentially useful for the formulation of injection that requires targeting for melanoma as well as sustained release at the site.

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Experimental Study for Innervation of Scalp and Face with WGA-HRP Method (WGA-HRP법을 이용한 두피와 안면부의 신경지배에 관한 연구)

  • Kang, Jun-Goo
    • The Korean Journal of Pain
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    • v.7 no.2
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    • pp.238-241
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    • 1994
  • Aim of this study was to discover the projection area of the first cervical spinal nerve. Subcutaneous injection of wheat germ agglutinin-horseradish peroxidase(WGA-HRP) was done at five points of young dogs scalp and face. After two days of survival time, animals were sacrificed by perfusion through the left ventricle of the heart. Trigeminal ganglion, first and second cervical dorsal root ganglion, superior cervical ganglion, middle cervical ganglion and stellate ganglion were removed. Projection area of wheat germ agglutinin-horseradish peroxidase in vestigated into above ganglions. Projection into the first cervical dorsal root ganglion and stellate ganglion was not found. This experiment is deemed valuable for the study of neuronal connection on the central nervous system.

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The Effect of Pretreatment with Various Mutagens on Glycoconjugates of Plasma Membrane in HeLa Cells (HeLa 세포에서 원형질 막의 glycoconjugate에 대한 수종 돌연변이원의 전처리 효과)

  • Lee, Jong-Hwa;Oh, Kyu-Seon;Lee, Dong-Wook;Shin, Eun-Joo;Um, Kyung-Il
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.116-122
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    • 1998
  • The present investigation has been performed to elucidate the effect of pretreatment with low dose of ultraviolet radiation (UV), ethyl methansulfonate (EMS), and bleomycin (BLM) on cell survival and lectin-binding glycoconjugates of plasma membrane in HeLa cells treated with mutagen. The percentage of survival of cells pretreated with 1 mM EMS following treatment with 10 mM EMS was higher than that of cells treated with 10 mM EMS alone. Wheat germ agglutinin (WGA) staining intensity of cells pretreated with 1 mM EMS and subsequently treated with 10 mM EMS was stronger than that of cells treated with 10 mM EMS alone. But, succinylated wheat germ agglutinin (sWGA) staining intensity of cells pretreated with 1 mM EMS and subsequently treated with 10 mM EMS was similar to that of cells treated with 10 mM EMS alone. These results suggest that the acquired resistance to EMS is related to the glycoconjugates containing sialic acid of plasma membrane involved in multidrug resistance or adaptive response in HeLa cells.

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Identification of Species-Specific Components between Hanwoo and Holstein Meat (한우 및 홀스타인육의 품종간 특이성분의 검색에 관한 연구)

  • 황보식;이수원;임태진;정구용
    • Food Science of Animal Resources
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    • v.21 no.3
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    • pp.246-255
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    • 2001
  • Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of muscles extracted with distilled water, saline solution, SDS or Trition X-100 showed simular protein patterns between Hanwoo and Holstein meat, indicating that SDS-PAGE technique may not be useful for the identification between Hanwoo and Holstein meat. Lectine blot analysis of muscle extracted with distilled water demonstrated that Hanwoo and Holstein meat had similar affinities for concanavalin A (Con A), ricinus communis agglutinin (RCA-120), ulex europaeus agglutinin (UEA-1) or peanut agglutinin (PNA) lectins. However, approximately 32.1 kDa component of Hanwoo meat showed high affinity for dolichos biflorus agglutinin (DBA) lectin. On the contrary, high molecular weight components of Holstein meat had the specific affinity for wheat germ agglutinin (WGA) lectin. Hanwoo meat-specific components were observed by lectin staining of heat-denatured meat at 100$^{\circ}C$ for 30 sec. Also, the component of heat-denatured meat at 100$^{\circ}C$ for 30 sec, which was slightly smaller than Hanwoo meat-specific component, was concentrated specifically in Holstein meat.

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Analysis of the Formation of Protoplasts and Regeneration of Cells in Phycomyces blakesleeanus

  • Joe, Fukui;Choi, Kwan-Sam;Atsushi Miyazaki;Tamotsu Ootaki;Taneaki Oikawa
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.72-78
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    • 2001
  • It is possible ot prepare protoplasts of the zygomycete fungus, Phycomyces blakesleeanus, by digesting the cell wall of spore germlings with commercially available chitinase and chitosanase. However, the cells without any cell walls immediately form large aggregates, and thus, it is difficult to isolate the individually separated protoplasts. Inherent problem with the formation of aggregates in preparing protoplasts could be solved by the use of bovine serum albumin (BSA). As a result, we were able to prepare a large number of single protoplsts quickly and easily. We took time-lapse photomicrographs of the formation of protoplasts, and found that there were certain regions of the cell wall of spore germlings that were sensitive to chitinase and chitosanase, although the cell wall of the original spores is known to be insensitive to these enzymes. There are two kinds of cell walls on a spore germling; one with a bound wheat germ agglutinin (WGA), and the other a bound concanavalin A (ConA). Furthermore, only cells with walls which had bound WGA were able to regenerate, while those with walls with bound ConA were not able to regenerate.

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Lectine-binding patterns of spermatogenic cells in the Jindo dog (진도견 정자형성계 세포들의 Lectin-binding patterns)

  • Park, Young-seok;Lee, Seong-ho
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.531-539
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    • 1996
  • The lectin-binding patterns in the testis of the sexually matured Jindo dog were investigated to study the distribution of glycoconjugates in the seminiferous tubule under light and transmission electron microscopy. Positive reactions to Wheat germ agglutinin(WGA) and Dolichos biflorus agglutinin (DBA) were observed in the Sertoli cell and in the residual body of spermatid with a stronger reaction in the Sertoli cell to the lectins than in the residual body. Strong reactions to Soybean agglutinin(SBA) and Peanut agglutinin(PNA) were observed in the acrosome vesicles of the Golgi- and cap-phase spermatid, while a moderate reaction was observed in the acrosome-phase, maturation-phase spermatid and the residual body. The acrosome area of the spermatid reacted intensively to Griffonia simplicifolia agglutinin( GS-I) when the cell was in the acrosome-phase and maturation-phase, and the same reaction to the GS-I was observed in the residual body. However, the seminiferous tubule did not react to Ulex europeus agglutinin I(UEA-I). The gold-labelling of the Sertoli cells with DBA resulted in positive reactions of the Sertoli cell column and processes when observed under the electron microscopy, while the Golgi-, cap- and acrosome-phase spermatids reacted positively to SBA in the peripheral low-dense area of the acrosome vesicle of spermatid. Based on these results, we concluded that differences in the lectin-binding pattern of the seminiferous tubules were recognized in the Jindo dog compared to other animals.

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Regional Difference in Distribution of Glycoconjugates in Mouse Epididymis (생쥐 부정소 부위별 당쇄 분포의 차이)

  • 계명찬
    • Development and Reproduction
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    • v.5 no.2
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    • pp.167-173
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    • 2001
  • To characterize the difference in glycoconjugates of mouse epididymis, lectin labeling of the tissue section was conducted using Ulex europaeus agglutinin I(UEA I), succinylated wheat germ agglutinin(sWGA), and Griffonia simplicifolia lectin-I(GSL-I). UEA I which binds to outer $\alpha$-L-fucose residue that is a terminal sugar of the side chain branched from oligosaccharide chain gave the labeling in the proximal caput epithelia exclusively. Lumen was commonly labeled in all of the organ. It suggested that the glycoconjugates bearing outer $\alpha$ -L-fucose residue were largely expressed in the initial segments ot epididymis and subjected to secretion. GSL-I which binds to terminal $\alpha$ -D-galactosyl residue of glycoconjugates gave the labeling in the cytoplasm of clear cells and basal cells, and cilia in corpus and cauda regions but not in the caput region. There was no vast difference in labeling pattern by sWGA which binds to N-acetyl-glucosamine residue among the epididymal regions. Clear cells in corpus and cauda epithelia showed more intense labeling by sWGA compared to principal cells, suggesting the functional specialization of this type of cells. The labeling intensities of luminal content by UEA I and sWGA decreased in cauda region compared to corpus region suggesting the presence of enzymatic activities responsible for processing the $\alpha$-L-fucose and N-acetyl-glucosamine residues from secreted glycoconjugates. In summary, the difference in glycoconjugates bearing the $\alpha$-L-fucose, $\alpha$-D-galactose, and N-acetyl-glucosamine residues according to the type of epithelial cells and epididymal segments suggests functional specialization and different roles of each segment in the processing of sperm surface antigens during the epididymal transit.

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Histochemical study on the tracheal development in fetuses and neonates of Korean native goats (한국재래산양의 태아 및 신생아 기관의 조직발달에 관한 조직화학적 연구)

  • Kim, Chong-sup
    • Korean Journal of Veterinary Research
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    • v.37 no.2
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    • pp.235-244
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    • 1997
  • The present study was performed to investigate the patterns of appearance of lectin in trachea of fetuses of 60, 90 and 120 days old and neonates of Korean native goat. Carbohydrate markers were used in histochemistry for the determination of the lectin by staining of avidin-biotin-peroxidase complex(ABC), and the markers consisted of biotin-labeled concanavalin A(Con A), dolichos biflorus agglutinin(DBA), rincinus communis agglutinin(RCA-I), ulexeuropalus communis agglutinin(UEA) and wheat germ agglutinin(WGA). 1. The Con A-binding reactions appeared moderately on the apical surface of the tracheal epithelia in 60 days old fetuses, and the reactions were similar on the tracheal epithelia and glands in 90 and 120 days old fetuses and neonates. 2. Reaction of the DBA appeared as the strongest meanwhile the DBA-binding reactions were determined strongly on the apical surface of the tracheal epithelia in the 60 days old fetuses. Reaction for the DBA on the tracheal epithelia and glands of 90 and 120 days old fetuses and neonates were in same manner. 3. The RCA-I-binding reactions appeared very strongly on the apical surface of the tracheal epithelia in 60 and 90 days old fetuses. Reaction to the RCA-1 appeared moderately on certain apical surface of tracheal epithelia and glands in 120 days old fetuses and neonates. 4. No reactions provoked for the UEA in trachea of 60 days old fetuses and neonates, but the UEA-binding reactions appeared moderately in the tracheal epithelia of 90 days old fetuses and weakly in 120 days old fetuses. 5. The WGA-binding reactions appeared very strongly on the apical surface of the tracheal epithelia in 60 and 90 days old fetuses, and moreover, the reactions were determined on the luminal surface of the tracheal gland in 90 days old fetuses. On the other hand, goblet cells of the tracheal epithelia and glands in neonates reacted moderately to the WGA.

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A morphological study of vomeronasal organ of Korean black goat (Capra aegagrus hircus) (한국흑염소 보습코기관의 형태학적 관찰)

  • Park, Changnam;Yang, Wonjun;Bae, Yeonji;Lee, Yongduk;Kang, Wanchoul;Ahn, Meejung;Shin, Taekyun
    • Korean Journal of Veterinary Research
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    • v.53 no.1
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    • pp.55-60
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    • 2013
  • The vomeronasal organ (VNO) plays an important role in reproduction and social activities in ruminants including goats. A morphological study on the structure of VNO and its epithelial cells was carried out in Korean black goats. Grossly, the VNO of Korean goats opens into mouth through incisive ducts. Microscopically, the epithelium of VNO consisted of medial sensory epithelium and lateral non-sensory epithelium. Several blood vessels and nerve bundles were observed in the lamina propria encased by vomeronasal cartilage. Immunohistochemical staining showed that protein gene product (PGP) 9.5 was immunostained in the receptor cells of the sensory epithelium and in some cells of the non-sensory epithelium. Galectin-3 was mainly observed in the supporting cells of sensory and non-sensory epithelium. Lectins including wheat germ agglutinin, Ulex europaeus agglutinin, Bandeiraea simplicifolia lectin Isolectin B4, Dolichos biflorus agglutinin and soybean agglutinin used in this study were bound in VNO sensory, non-sensory epithelia as well as in the lamina propria with varying intensity. Collectively, this is a first descriptive morphological study of VNO of Korean black goat with special reference to lectin histochemistry.

Histochemical study of lectin-binding patterns in the rat vomeronasal organ during postnatal development

  • Lee, Wonho;Ahn, Meejung;Park, Changnam;Taniguchi, Kazumi;Moon, Changjong;Shin, Taekyun
    • Korean Journal of Veterinary Research
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    • v.52 no.1
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    • pp.1-8
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    • 2012
  • Histochemical patterns of lectin binding during development of the rat vomeronasal organ (VNO) were studied to determine whether glycoconjugates are differently expressed after birth. Three types of lectins, Dolichos biflorus agglutinin (DBA), wheat germ agglutinin (WGA), and Ulex europaeus agglutinin I (UEA-I), were studied histochemically in the rat VNO at various stages post-birth: postnatal days 1 and 7, the preweaning period (4 weeks after birth), and at sexual maturity (8 weeks after birth). The free border of the vomeronasal sensory epithelium was positive for both WGA and UEA-I in rats of all ages; whereas, VNO receptor cells and supporting cells were positive only for both WGA and UEA-I from 4 weeks after birth. DBA reactivity was detected in the free border but less so in receptor cells and supporting cells. WGA and UEA-I, but not DBA, showed similar patterns in various ages. In the Jacobson's gland, WGA, UEA-I and DBA were detected in some acini from 4 weeks after birth but not at postnatal days 1 or 7. Collectively, reactivity for three lectins, WGA, UEA-I and DBA, increased in receptor cells and gland acini during postnatal development, possibly contributing to the enhanced chemoreception in rats.