• Journal of Life Science
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• v.18 no.6
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• pp.782-788
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• 2008

The aim of this work is to show differences in the pattern of glycoconjugate composition in the intestines of four teleostean species (Sebastes schlegeli, Halichoeres poecilopterus, Bryzoichthys lysimus, and Takifugu pardalis). We compared four regions of all species studied. The specimens were processed and stained with nine kinds of biotinylated lectins (DBA, SBA, PNA, BSL- I , RCA- I , sWGA, UEA- I , LCA and Con A). Except for Sebastes schlegeli, no differences between regions were observed. The intestinal epithelium of Halichoeres poecilopterus possessed D-glucose/mannose residues in all regions. ${\beta}$-N-acetyl-D-galactosamine was distinctive along the intestines, although the pattern of diversity was different in Sebastes schlegeli, Bryzoichthys lysimus, and Takifugu pardalis. Additionally, the occurrence of Galactose-${\beta}$-1,3-N-acetyl-D-galactosamine and ${\alpha}$-D-galactose were confirmed in the proximal, middle, and distal intestine of Sebastes schlegeli, while rectal intestine lacked these sugar residues. Along with ${\beta}$-N-acetyl-D-galactosamine, ${\beta}$-N-acetyl-D-glucosamine and D-glucose/mannose were also determined in Bryzoichthys lysimus. Galactose-${\beta}$-1,3-N-acetyl-D-galactosamine, D-galactose, and D-glucose/mannose were also present in Takifugu pardalis.

• The Korean Journal of Zoology
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• v.38 no.4
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• pp.514-522
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• 1995

In order to examine the interaction of albumin with the sperm during capacitation in mouse, proteins of cauda epididymal sperm were extracted under various conditions and analyzed with SDS-PAGE. Sperm surface labeling patterms were also examined using fluorochroin~conjugated wheat germ agglutinin (WGA) and bovine serum albumin (BSA). Albumin was detached from the sperm surface during the incubation and seemed to be constituted the major protein components of the conditioned media in which sperm incubated for 90 mm. Detachment of albumin from the sperm was not affected by the Ca2+ in the medium. WGA-FITC labeling confirmed that Triton X-100 permeabilired plasma membrane overlaying the apical segment of sperm head and detached plasma membrane associated proteins having negatively charged glycoconjugates. BSA-FITC labeling of epididymal sperm occurred on the apical segment of periacrosoinal region and postacrosomal region of the head. BSA-FITC labeling was not observed in periacrosoinal region of the sperm treated with Ca2+-ionophore ~3187 (10 MM)~ whereas the postacrosome region of acrosome-reacted sperm was still labeled after the AR. These results suggest that albumin bound to the surface of epididymal sperm is detached during the capacitation process, and it might be involved In physiological change of sperm plasma membrane accompanying the capacitation.

• Korean Journal of Environmental Agriculture
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• v.30 no.4
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• pp.390-394
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• 2011

BACKGROUND: Plant expression system for mass production of recombinant proteins has several advantages over other existing expression systems with economical and safety issues. Breast cancer is a cancer originating from breast tissue and comprises almost 25% of all cancers in women world widely. Lewis-Y antigen is difucosylated oligosaccharide and is carried by glycoconjugates at cancer cell surface. In this study, the anti-breast cancer mAb BR55, which recognizes the epitope Lewis-Y, was expressed in the plant expression system. METHODS AND RESULTS: We have developed plant system for production of mAb BR55 with or without KDEL (the ER retention signal). This ER retention signal was attached to C-terminus of protein to help retain the recombinant glycoprotein carrying oligomannose glycans and enhance glycoprotein accumulation. PCR analysis was performed and confirmed the presence of recombinant genes. Western blot validated that the recombinant proteins mAb BR55 with or without KDEL were expressed in transgenic plants, moreover, the expression level of the mAb BR55 with KDEL was higher compared to the mAb BR55 without KDEL. CONCLUSION: These results indicate that KDEL fusion is a good way to produce proteins and plant can be an ideal expression system to obtain proteins and enhance accumulation of proteins.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.4
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• pp.270-284
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• 2021

The vomeronasal organ (VNO) is critical for reproduction and social behavior in ruminants, including cattle. The present study examined the structure of the VNO and its epithelial cells in neonatal and adult Korean native cattle (Hanwoo), Bos taurus coreanae, using immunohistochemistry and lectin histochemistry. Histologically, the VNO comprised two types of epithelia: medial vomeronasal sensory (VSE) and lateral vomeronasal non-sensory epithelia (VNSE). Numerous blood vessels and nerve bundles were observed within the vomeronasal cartilage encased lamina propria. Immunohistochemistry revealed high expression level of protein gene product9.5 and moderate expression level of olfactory marker protein in the neuroreceptor cells of the VSE and occasionally in some ciliated cells of the VNSE in both neonates and adults. The properties of the glycoconjugates in the VNO were investigated using 21 lectins, most of which were expressed at varied intensities in the VSE and VNSE, as well as in the lamina propria. Several lectins exhibited variations in their intensities and localization between neonatal and adult VNOs. This study is the first descriptive lectin histochemical examination of the VNO of Korean native cattle with a focus on lectin histochemistry, confirming that the VNO of Korean native cattle is differentiated during postnatal development.

• Journal of Life Science
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• v.16 no.6
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• pp.1014-1028
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• 2006

This study attempted to investigate the developmental alterations of the cerebral cortex. The effects of EGEE on the developmental cerebral cortex in the prenatal, postnatal and adults were examined by lectin histochemical methods. To investigate the change of sugar residues of glycoconjugates, biotinylated lectins(DBA, SBA, PNA, BSL-1, RCA-1, sWGA, UEA-1, Con A and LCA) were detected with by IHC using ABC kit. In the case of injection of EGEE, the reactions to Con A and LCA were shown in binding phase in the cerebral cortex commonly, and the reactions to PNA, RCA-1 and LCA were shown partially, the number of lectins to be shown reaction were decreased, and there were no reactions to DBA, SBA, BSL-1, RCA-1 and UEA-1. The reaction to Con A was similar to control group during developmental phases. The reaction to LCA was increased in the fetal, suckling, and weanning phases compared with control group. But there were no reactions to SBA and sWGA, the reaction to PNA was decreased in the frontal and occipital cortex and no reaction to sWGA in the fetal phase. There were no reactions to sWGA and PNA in the suckling phase and, no reaction to PNA and sWGA. The reaction to Con A was decreased in the frontal, parietal and occipital cortexes and, the reaction to LCA was decreased in the frontal and occipital cortexes in adult phase. As the results, the effects of EGEE, environmental hormone on the each part of cerebral cortex have shown differences. But, It had deep effect on the differentiation and growth in the cerebral cortex pathologically. In particular, the effect was severe in suckling phase. $Galactosyl-({\beta}-1,3)-N-acetyl-D-galactosamine$,${\beta}-N-acetyl-D-galactosamine$ and ${\beta}-N-acetyl-D-glucosamine$ were decreased while ${\alpha}-D-mannose$ and ${\alpha}-D-glucose$ were increased. It affected the sugar metabloism, and it was severe in fetal and suckling phases.

• Development and Reproduction
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• v.2 no.2
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• pp.149-155
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• 1998

The polysialic acid (polySia) glycotope covalently modifies cell surface glycoconjugates on cells as evolutionarily diverse as microbes and human. The recent chemical identification of polysialylated glycoproteins in the jelly coat and on the cell surface of the sea urchin egg raises important questions about their biosynthesis and possible function. Using CMP-[$^{14}$ C]Neu5Ac as substrate and cell free preparations from eggs and embryos of the sea urchin Stronglylcentrotus nudus, we have identified a membrane associated CMP-Neu5Ac:poly-$\alpha$2, 8 sialosyl sialyltransferase (polyST) that transfers Neu5Ac to an endogenous acceptor. Optimal conditions for the polyST activity were found to be 2$0^{\circ}C$ in 20 mM MOPS buffer (pH 7.0). The polyST activity was increased 2.7 times by the addition of 10 mM $Mg^2$$^{+}$. The membrane-associated polyST also catalyzed the polysialylation of mammalian ganglioside GD3. Given that no structurally similar natural polysialylated gangliosides have been described, nor were observed in the present study, we conclude that a single polyST activity catalyzes sialylation of the endogenous acceptor and the gangliosides. Using an excess of GD3 as an exogenous acceptor, it was established that the expression of the polyST in S. nudus embryos increased rapidly at the mesenchyme blastula stage and reached at maximum at the gastrula stage. The finding that this polyST in the sea urchin embryo is developmentally regulated raises the possibility that it may play a role in the changing cell and tissue interactions that occur during gastrulation and the early stages of spicule formation.n.

• Journal of Life Science
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• v.14 no.6 s.67
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• pp.1009-1017
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• 2004

Sialyltransferases cloned so far show the remarkable tissue-specific expression, which is correlated with the existence of cell type-specific sialylated sugar structure in glycoconjugates. In the previous studies, we found various mRNA isoforms of human sialyltransferases generated by alternative splicing and alternative promoter utilization. To understand the regulatory mechanisms for specific expression of human sialyltransferase genes and for production of their mRNA isoforms, in this study, we have isolated and characterized five kinds of human sialyltransferase genes: hST3Gal II, hST8Sia II, hST8Sia III, hST8Sia IV, and hST8Sia V. The hST3Gal II gene is composed of six exons, which span over 17kb, with exons ranging in size from 46 to over 1017 bp. The hST8Sia III gene comprises over 10 kb, and consists of only four exons, which is much smaller and simpler than other human sialyltransferase genes. In contrast, three genes (hST8Sia II, hST8Sia IV and hST8Sia V) span more than 70 kb, and comprise five or more exons. All exon-intron boundaries follow the GT-AG rule. In particular, the sialylmotif L, which is a highly conserved region in all cloned sialyltransferases, was found in one exon of hST8Sia III, whereas this motif is encoded by discrete exons in the other human sialyltransferases. Exon structures of these sialyltransferase genes show the structural diversity, as found in other human sialyltransferase genes reported so far. We determined the transcription start site of hST3Gal II gene by the 5'-RACE and cap site hunting experiments.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.3
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• pp.179-186
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• 2002

Objective: Lectins are cell-agglutinating and sugar specific proteins or glycoproteins of non-immune origin that precipitate glycoconjugates having saccharides of appropriate complementarity. Because of these properties, plant lectins have been used to help characterize the carbohydrate moieties of glycoproteins in the zona pellucida (ZP) of several mammalian species including pigs. Treatment of oocytes with various lectins blocks sperm binding to the ZP in various mammalian species. This study was undertaken to examine the distribution of sugar residues in the ZP of pig oocytes matured in vitro and the ability of spermatozoa to bind to ZP and in vitro penetration in oocytes treated with fluorescein isothiocyanate (FITC)-labelled lectins. Materials and Methods: The lectins of Banderiaea simplicifolia (BS-II, bind to $\beta$-D-N-acetylglucosamine), Canavalin ensiformis (Con A, bind to $\alpha$-D-Mannose), Lens culinaris (LCA, bind to a-D-Mannose), Ricinus communis (RCA-I, bind to $\beta$-D-Galactose) and Ulex europaeus (UEA-I, bind to $\alpha$-L-Fucose) were examined for spermatozoa penetration, binding capacity to ZP and distribution of lectins. Results: The penetration rates were significantry (p<0.05) higher in control oocytes (63%) than those treated with all lectins, but penetration rates ($40{\sim}49%$) were simililar in group treated with lectins. The incidence of monospermy was similar in oocytes untreated and UEA-I, but it was higher in oocytes treated with BS-II, Con A, RCA-I and LCA. The porcine oocytes cultured for 48 h in TC-199 medium were freed from cumulus cells and treated for 30 min with fluorescein isothiocyanate-labelled lectins. When examined under fluorescein illumination, higher (p<0.001) proportions of oocytes showed fluorescein of zona pellucida after treatment with Con A (93%), LCA (93%) and RCA-I (100%) than BS-II (37%) and UEA-I (50%). All of the oocytes treated with RCA-I exhibited strong fluorescein in the outer region of the zona pellucida while those treated with LCA exhibited strong fluorescein throughout the zona pellucida. BS-II bounded mainly to the outer region and UEA-I bounded mainly to the inner region of the zona pellucida, with either strong or weak fluorescein. At 120 min after insemination in vitro, fewer spermatozoa were bound to the zona pellucida of the oocytes treated with BS-II, Con-A and RCA-I. Of the lectins, Con A most inhibited sperm binding. Conclusions: These results suggest that $\beta$-D-Galactose residues in the porcine zona pellucida may act as primary sperm receptors and inducers of the sperm acrosome reaction and these sugar residues may be involved in the block to polyspermy.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.23 no.2
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• pp.118-151
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• 1997

Allergic contact dermatitis is a common skin disease resulting from specific immumologic sensitization to topically applied various allergen. Purpose of this study was to investigate skin morphologic chganges by light microscopic, changes of protein band by SDS-PAGE in the skin treated on the surfactant. Furthermore, lectin histochemistry is used to know the chagnges of the terminal sugar of the glycoconjugate in the skin treated on the surfactant. The results were as follows : The 1 day of treated group was indicated the enlargement of capillary, the 3 days of treated group was showed that the cytoplasm was eosinophlic by the pyknotic of nucleus. The 6 days of treated group was observed hyperkeratinoid, increased of inflammation cells to epithelium. The 9 days of treated group was appeares that serum crust was sheded and formed a acanthosis. But dermis layer was cytoplasmic vaculation and enlargement of intercellular space. The 12 days treated group made up new epidermis layer of seven layer and of observed an decreased in quantity of inflammation cells. The mast cell of degranulated type was increased in treated surfactnat. It was secreating granules hold histamin, serotonin and heparin. Accordingly, protein band of electrophortic phase was observed a sudden changes since 3 days treated group. At observeation of the cell-surface glycoconjugates, LCA, PNA, SBA and WGA lectin positive cells but inducated lectin negative cells in Con-A. Furthermore, PNA and SBA showed stroger positive reaction as treated surfactant group.

• Journal of Life Science
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• v.23 no.12
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• pp.1541-1552
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• 2013

The purpose of this study was to investigate the structure and secretory function of the von Ebner's gland in parasympathetic or sympathetic nerve innervation. Sprague Dawley rats were sacrificed 3, 7, 10, 14, and 21 days after bilateral glossopharyngeal or hypoglossal nerve axotomy, respectively. The circumvallate papilla portion of the tongue was dissected and we observed morphological changes in the von Ebner's gland. The properties of glycoconjugate in the von Ebner's gland were investigated using nine biotinylated lectins (PSA, UEA I, GSL I $B_4$, ECL, DBA, SBA, HPA, SJA, or sWGA). Compared with the control group, cytoplasmic vacuoles appeared in the serous acini of the von Ebner's gland in the 3-day group, and the serous acini were significantly vacuolized and degenerated in the 10-day group after glossopharyngeal nerve axotomy. However, the structure of the von Ebner's gland did not change after hypoglossal nerve axotomy. In the control group, the von Ebner's glands secreted glycoconjugates containing ${\alpha}$-D-galactose, N-acetyl-D-galactosamine, and N-acetyl-D-glucosamine oligomer, and the amount of the secretion decreased significantly in the 10-day group after glossopharyngeal nerve axotomy. However, the amount of the glycoconjugate secretion did not change after hypoglossal nerve axotomy. Therefore, the results of this study suggest that the glossopharyngeal nerve containing parasympathetic nerve fibers is important for maintaining the structure of and secretory function in the von Ebner's gland in rats.