• Title/Summary/Keyword: lectin staining

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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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Separation of Lentil Lectin Using Free-Flow Electrophoresis (자유유동 전기이동을 이용한 Lentil Lectin의 분리)

  • 류화원;이동일장호남
    • KSBB Journal
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    • v.9 no.2
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    • pp.115-121
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    • 1994
  • A Purification device with 30-channel free-flow electrophoresis was assembled to treat samples of 240m1 volume for purification of lentil lectin (LcH) from lentil seeds with no impurities in a silverstained PAGIEF gel. HEPES(50mM)-Ttis(50mM), Cycloserine(50mM)-urea(3M), Histidine(50mM)-urea(3M) were used as ampholytea among which Histidine(50mM)-urea(3M) (pI 7.65) was found best in resolution. LcH is known to be present in the form of LcH-A, LcH-B and the complex of the two. The complex, however, disappeared when urea was added in the ampholytes, which suggested that the complete purification of two isolectins is possible using the present multistep purificaton device.

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Comparative lectin binding patterns of Cochlodinium polykrikoides Margalef

  • Rhodes, Lesley L.;Cho, Yong-Chul;Cho, Eun-Seob
    • Journal of the korean society of oceanography
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    • v.35 no.3
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    • pp.153-157
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    • 2000
  • Four different FITC-conjugated lectins were used to visually evaluate lectin binding activity by optical staining quality using confocal laser scanning microscopy (CLSM) of Cochzodinium polykrikoides in nature (wild type) and culture (cultured type). Cells from the field and cultures treated with ConA fluoresced only at the outer cell wall, and the abundance and distribution of the fluorescent signal were similar. Treatment with PWM and HPA did not elicit fluorescence at the cell surface, but the wild type exposed to HPA showed greater binding than did the cultured cells, possibly due to greater concentrations of glucosamine. The wild type cells treated with LBL lectin showed a strong green fluorescence on the cell surface, whereas cultured cells did not. Signal intensity and abundance were greater than for any other lectins tested in this study. These results suggest that wild type and cultured type are significantly different based on surface sugar production. In particular, the wild type cells apear richer in galactosamine-like moieties. Neither glucose nor mannose-like moieties were present in either wild types or cultured cells.

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Isolation and Characterization of Lectin in Soybean(Glycine max L.) (대두(Glycine max L.)의 렉틴 분리 및 특성)

  • 박원목;이용세;박상호;김성환;윤경은
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.34 no.2
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    • pp.120-126
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    • 1989
  • This experiment was carried out to investigate the lectin of soybean (Glycine max L.) seed. Purification was done by 50-80% ammonium sulfate precipitation, CM-cellulose and Sephadex G-100 column. The purity was ascertained by electrophoresis. The molecular weight of purified lectin was estimated as 132,000. It was composed of three subunits which molecular weight was 45,000. The lectin was identified as glycoprotein by Schiff's reagent staining and Dubois method. The lectin agglutinated erythrocytes of rabbit and human. The amounts of the lectin to agglutinate human erythrocytes differed among the blood types: The blood type A required the least amount, the next was B, O, and AB in order. The agglutination was specifically inhibited by 5${\mu}$g/ml of N -acetyl.-D-galactoseamine and 200${\mu}$g/ml of D-galactose. Other tested sugars could not inhibit the agglutination of the erythrocytes by the lectin.

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Characterization and Distribution of Glycoconjugates in Human Pulmonary Tubercles by Lectin Histochemistry (폐결핵 결절에서 복합당질의 분포에 관한 Lectin 조직화학적 연구)

  • Yoon, Sik;Kim, Ji-Hong;Shin, Cheol-Shik;Jeong, Suk;Son, Mal-Hyun;Song, Sun-Dae;Kim, Jin-Jeong
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.3
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    • pp.248-261
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    • 1994
  • Background: Lectins are proteins or glycoproteins of non-immune origin that recognize a specific sequence of sugar residues. The availability of a large number of lectins has provided the capacity to identify selectively glycoconjugates possessing distinctive chemical structure in diverse sites of highly specialized biological activity. The purpose of the present study was to investigate the lectin binding patterns of various components in human pulmonary tubercles. Method: Biopsy specimens of tuberculous lung were obtained from male adult patients who underwent a surgical resection for severe pulmonary tuberculosis. The specimens were processed and stained with 13 kinds of biotinylated lectins according to some modification of Hsu and Raine's methods. Results: 1) In the caseous necrotic lesions, BS $I-B_4$ showed negative reaction and BS I were also negative except some irregularly-shaped cells located in the marginal zone. All other lectins, however, showed a positive reaction with various binding patterns. 2) The epithelioid cells were broadly divided into three groups according to the reaction patterns in the cytoplasms and cell membranes. 3) WGA, ECL, PHA-L, PHA-E and LCA showed strong staining in the lymphocytes. 4) SBA showed a different binding patterns between the endothelial layers located in the region beyond the fibrous layers and those located within the fibrous layers. 5) PNA showed a positive reaction in the outer 1/3 to 1/2 of the fibrous layer, but showed no staining in the inner 1/2 to 2/3 of the fibrous layers. Conclusion: The present lectin histochemical study provided a useful information to assess the characterization and distribution of various glycoconjugates in each constituent of human pulmonary tubercles. The results demonstrate structural differences in the glycoconjugate composition of various components of the tubercles and reveal changes in glycosylation in the components during soft tubercle formation. This study provides a new data useful for the studies on the pathogenesis and pathology of human pulmonary tubercles.

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Sun Ginseng Protects Endothelial Progenitor Cells From Senescence Associated Apoptosis

  • Im, Woo-Seok;Chung, Jin-Young;Bhan, Jae-Jun;Lim, Ji-Yeon;Lee, Soon-Tae;Chu, Kon;Kim, Man-Ho
    • Journal of Ginseng Research
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    • v.36 no.1
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    • pp.78-85
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    • 2012
  • Endothelial progenitor cells (EPC) are a population of cells that circulate in the blood stream. They play a role in angiogenesis and, therefore, can be prognostic markers of vascular repair. Ginsenoside $Rg_3$ prevents endothelial cell apoptosis through the inhibition of the mitochondrial caspase pathway. It also affects estrogen activity, which reduces EPC senescence. Sun ginseng (SG), which is heat-processed ginseng, has a high content of ginsenosides. The purpose of this study was to investigate the protective effects of SG on senescence-associated apoptosis in EPCs. In order to isolate EPCs, mononuclear cells of human blood buffy coats were cultured and characterized by their uptake of acetylated low-density lipoprotein (acLDL) and their binding of Ulex europaeus agglutinin I (ulex-lectin). Flow cytometry with annexin-V staining was performed in order to assess early and late apoptosis. Senescence was determined by ${\beta}$-galactosidase (${\beta}$-gal) staining. Staining with 4'-6-Diamidino-2-phenylindole verified that most adherent cells (93${\pm}$2.7%) were acLDL-positive and ulex-lectin-positive. The percentage of ${\beta}$-gal-positive EPCs was decreased from 93.8${\pm}$2.0% to 62.5${\pm}$3.6% by SG treatment. A fluorescence-activated cell sorter (FACS) analysis showed that 4.9% of EPCs were late apoptotic in controls. Sun ginseng decreased the apoptotic cell population by 39% in the late stage of apoptosis from control baseline levels. In conclusion, these results show antisenescent and antiapoptotic effects of SG in human-derived EPCs, indicating that SG can enhance EPC-mediated repair mechanisms.

A Comparative Study of Korean mistletoe lectin and bee venom on mechanism in inducing apoptosis of Hep G2, a liver cancer cell

  • Lim, Seong-Woo
    • The Journal of Korean Medicine
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    • v.39 no.4
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    • pp.158-170
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    • 2018
  • Objectives: The objective of this study is Korean mistletoe lectin (Viscum album coloratum agglutinin, VCA) and bee venom (BV) to experimental prove comparative study of VCA and BV on the anti-cancer effect and mechanisms of action. Methods: In this study, it was examined in a human hepatocellular carcinoma cell line, Hep G2 cells. Cytotoxic effects of VCA and BV on Hep G2 cells were determined by 3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide (MTT) assay in vitro. VCA and BV killed Hep G2 cells in a time- and dose-dependent manner. Results: The apoptotic cell death was then confirmed by propidium iodide staining and DNA fragmentation analysis. The mechanisms of action was examined by the expression of anti-apoptotic proteins and activation of mitogen-activated protein kinases. Treatment of Hep G2 cells with VCA activated poly (ADP-ribose) polymerase-1 (PARP-1) known as a marker of apoptosis, and mitogen-activated protein kinases signaling pathways including SAPK/JNK, MAPK and p38. BV also activated PARP-1, MAPK, p38 but not JNK. The expression level of anti-apoptotic molecule, Bcl-X, was decreased by VCA treatment but not BV. Finally, the phosphorylation level of ERM proteins involved in the cytoskeleton homeostasis was decreased by both stimuli. Conclusion: We examined the involvement of kinase in VCA or BV - induced apoptosis by using kinase inhibitors. VCA-induced apoptosis was partially inhibited by in the presence.

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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Histochemical Study on the Uterine Development in Fetues and Neonates of Korean Native Goats (한국재래산양의 태아 및 신생아 자궁의 조직 발달에 관한 조직화학적 연구)

  • 정순희;김종섭;정헌식;이규승;서길웅
    • Korean Journal of Animal Reproduction
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    • v.22 no.1
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    • pp.19-27
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    • 1998
  • The present study was performed to investigate the patterns of a, pp.arance of lectin in uterus of fetuses of 90 and 120 days old and neonates of Korean native goat. The carbohydrate markers were used in histochemistry for the determination of the lectin by staining of avidin-biotin-per-oxidase complex(ABC), rincinus communis agglutinin(RCA-I), ulexeuropalus communis agglutinin(UEA) and wheat germ agglutinin(WGA). 1. The effects of this study were as follows; 1. The binding reactions for Con-A were weak on the mucosal epithelium of endometrium in 90 and 120 days old fetuses, and neonates and moderate at the free surface of mucosal epithelia. 2. The binding reactions for DBA was partially moderate on the mucosal epithelium of endometrium and partially strong at the free surface of mucosal epithelia in 120 days old fetuses. In neonates, the reactions were strong on the mucosal epithelium and gland primordium of endometrium, and the secretions at the free surface showed strong reactions for DBA. But, in 90 days old fetuses, the reaction was negative. 3. The binding reactions for RCA-I were moderate on the mucosal epithelium of endometrium and at the free surface of mucosal epithelia in 90 days old fetuses. In 120 days old fetuses, the reactions were weak on the mucosal epithelium of endometrium and moderate at the free surface of mucosal epithelia. In neonates, the reactions were moderate on the mucosal epithelium of endometrium and strong at the free surface of mucosal epithelia and also strong in the uterine gland. 4. The binding reactions for UEA were negative in 90 and 120 days old fetuses and neonates. 5. In 90 days old fetuses, the binding reactions for WGA were generally weak on the mucosal epithelium of endometrium, but several epithelial cells showed moderate reaction for WGA. In 120 days old fetuses and neonates, the reactions were moderate on the mucosal epithelium and blood vessels of endometrium and strong at the free surface of mucosal epithelia.

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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.