A tuber lectin from Arisaema jacquemontii Blume belonging to family Araceae was purified by employing a single step affinity chromatography using column of asialofetuin-linked amino activated silica beads and the bound lectin was eluted with 100 mM glycine-HCl buffer pH 2.5. The purified A. jacquemontii lectin (AJL) showed a single protein band with an apparent molecular mass of 13.4 kDa when submitted to SDS-polyacrylamide gel electrophoresis under reducing as well as non-reducing conditions. The native molecular mass of AJL determined by gel filtration on a Biogel P-200 column was 52 kDa and its carbohydrate content was estimated to be 3.40%. Thus AJL is a tetrameric glycoprotein. The purified lectin agglutinated erythrocytes from rabbit but not from human. Its activity was not inhibited by any of the mono- and disaccharides tested except N-acetyl-D-lactosamine having minimal inhibitory sugar concentration (MIC) 25 mM. Among the glycoproteins tested only asialofetuin was found to be inhibitory (MIC $125\;{\mu}g/mL$). A single band was obtained in native PAGE at pH 4.5 while PAGE at pH 8.3 showed two bands. Isoelectric focusing of AJL gave multiple bands in the pI range of 4.6-5.5. When incorporated in artificial diet AJL significantly affected the development of Bactrocera cucurbitae (Coquillett) larvae indicating the possibility of using this lectin in a biotechnological strategy for insect management of cucurbits. Larvae fed on artificial diet containing sub-lethal dose of AJL showed a significant decrease in acid phosphatase and alkaline phosphatase activity while esterase activity markedly increased as compared to larvae fed on diet without lectin. Out of various human cancer cell lines employed in sulphorhodamine B (SRB) assay, this lectin was found to have appreciable inhibitory effect on the in vitro proliferation of HCT-15, HOP-62, SW-620, HT-29, IMR-32, SKOV-3, Colo-205, PC-3, HEP-2 and A-549 cancer cell lines by 82, 77, 73, 70, 41, 41, 37, 29, 21 and 21% respectively.
Kim, Ji Hye;Chung, Eun-Jin;Park, Hyun Kyung;Moon, Soo Ji;Choi, Su-Mi;Oh, Sung Hee
Clinical and Experimental Pediatrics
/
v.52
no.9
/
pp.1053-1058
/
2009
Cytomegalovirus (CMV) is one of the most commonly encountered viral pathogens in newborn infants and is found in 0.3-2.4% of all live births. It has been demonstrated that 40-96% of seropositive mothers shed the virus via their breast milk. Breast milk containing CMV can cause almost one-third of CMV infections occurring in infants. A case of postnatal CMV infection in an extremely premature infant (gestational age $24^{+5}$ weeks, birth weight 750 g) transmitted via breast milk is presented. For neonatal intensive care unit (NICU) management of severe thrombocytopenia, anemia, and sepsis syndrome, the infant received repeated transfusions of platelets; intravenous (IV) immunoglobulins; and gamma- irradiated, filtrated packed red cells and was fed her mother's breast milk since the second week of life. CMV infection was diagnosed with positive CMV immunoglobulin M (IgM) and positive urine CMV culture at the second month of life. Considering the negative CMV IgM and urine CMV culture at birth, postnatally-acquired CMV infection was suspected and confirmed with completely identical nucleotide sequence alignments of the infantile blood isolate and the maternal breast milk isolate. To our knowledge, this is the first case of proven postnatal CMV infection transmitted via breast milk in an extremely premature infant in Korea.
Adherence of Candida albicans(C. albicans) to the surface of a denture is believed to be an initial and essential step in the formation of denture-induced stematitis. Previous studies have provided enormous infomation on the relationship between composition of palatine gland/parotid saliva and upper denture stomatitis. Relatively little information is available on the correlation between lower denture stomatitis and sublingual-submandibular ( SLSM ) saliva. The plaque samples were collected from the two sites($100mm^2$) on the inner surface of lower partial denture corresponding to the stematitis and healthy region of the lower partial dentures of 12 denture stomatitis patients and 6 nor-mal persons who wore lower partial dentures. The samples were plated to isolate C. albicans on a selective Saboraud's dextrose agar plate and the isolates were identified by germ tube test and gram staining. The subjects were divided into group I (stomatitis with C. albican), group II (lesion without C. albicans), group III (no lesion but C. albicans), and group IV (normal and healthy denture wearer). Individual SLSM saliva($20{\mu}g$ of protein) was analyzed by SDS-PAGE (SDS -poly-acrylamide gel electrophoresis) with Coomassie brilliant blue and PAS(Periodic Acid Schinff) stain-ing. The salivary proteins separated in the polyacryamide gels were subjected to immunoblot anaysis using anti-lactoferrin, anti-sIgA, and anti-secretory component of sIgA. In this study using custom made acrylic denture resin beads(5mm in diameter) coated with stimulated individual SLSM saliva, the binding ability of individual C. albicans strains to the beads was observed. Levels of C, albicans adhered to the acrylic resin beads were determined by measuring the optical density of the bound C. albicans to the beads at 580nm. The results showed that a higher number of C. albicans was observed in the lesion site than healthy site. The saliva of group I contained more high molecular weight glycoprotein(mucin, MGI) as compared to group II, III and IV. And lactoferrin and sIgA affected to the binding ability of C. albicans to acylic resin beads. Binding ability of individual C. albicans to the acrylic resin coated with respective individual saliva was found to be greater in group I than the other 3 groups. And when bound cells of C. albicans isolated from individual subject #2 to the saliva coated beads were used binding ability of subject #2 saliva coated beads was founed to be greater than the other sutjects. These results suggested that denture induced stomatitis is related to individual patient's salivary protein composition, especially MG-1. Future studies will be directed toward saliva exam-ination of patients who have general disease and analysis of pellicles formed on prosthesis with respect to oral disease.
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
/
pp.344-344
/
2017
The glycoprotein known as glomalin-related soil protein (GRSP) is abundantly produced on the hyphae and spores of arbuscular mycorrhizal fungi (AMF) in soil and roots. GRSP play a decisive role in the soil aggregation, but GRSP was also sensitive to agricultural managements. Thus, our objectives were to assess the effect of different cover crop incorporation on the GRSP content in soil and growth of subsequent soybean and maize. Pot experiments with the incorporation of four cover crops were set up. The same amount (666g) of aboveground plant parts of wheat (AMF host), hairy vetch (AMF host), mustard (non-host) and rapeseed (non-host) was separately incorporated into soils. The aboveground plant parts and roots of soybean and maize were grown in each incorporated pots and sampled at 6 and 9 weeks after sowing. Our results showed that the different cover crops incorporation affected soil biological and chemical properties such as EC, $NO_3-N$ content, ${\beta}-glucosidase$ activity, alkaline phosphatase (ALP) activity and GRSP content. The soil EC and $NO_3-N$ content in the hairy vetch, mustard and rapeseed was higher compared to the wheat. The ${\beta}-glucosidase$ activity in the wheat and hairy vetch was significantly higher than that in the mustard and rapeseed, and the ALP activity in the wheat was significantly higher than that in the hairy vetch, mustard, and rapeseed. The GRSP content in the mustard and rapeseed was significantly lower than that of the hairy vetch and wheat. Moreover, The top dry weight and leaf area of soybean and maize in the hairy vetch at 6 weeks were significantly higher compared to the other treatments. Our results indicated that the incorporation of mustard and rapeseed may cause indirectly the decrease of GRSP content and soil enzyme activity in soil. One possible explanation for the decrease of GRSP in non-AMF host crop treatments may be the decrease of AMF density in the soil. AMF are not able to form a symbiotic relationship with Brassicaceae roots due to the release of anti-fungal compounds. This means the AMF may not be able to produce GRSP in the soil. However, the differences in the benefit of cover crop incorporation were shown only by a pot experiment. Comparative investigations of crop residue managements would be applied to both pot experiment and field study to clarify a better selection of cover crops in rotation to encourage GRSP production.
Kim, Jeong-Min;Byambaragchaa, Munkhzaya;Kang, Myung-Hwa;Min, Kwan-Sik
Development and Reproduction
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v.22
no.2
/
pp.143-153
/
2018
The large extracellular domain of glycoprotein hormone receptors is a unique feature within the G protein-coupled receptors (GPCRs) family. After interaction with the hormone, the receptor becomes coupled to Gs, which, in turn stimulates adenylyl cyclase and the production of cAMP. Potential phosphorylation sites exist in the C-terminal region of GPCRs. The experiments described herein represent attempts to determine the functions of the eel follicle-stimulating hormone receptor (eelFSHR). We constructed a mutant of eelFSHR, in which the C-terminal cytoplasmic tail was truncated at residue 614 (eelFSHR-t614). The eelFSHR-t614 lacked all potential phosphorylation sites present in the C-terminal region of eelFSHR. In order to obtain the eelFSHR ligand, we produced recombinant follicle-stimulating hormone ($rec-eelFSH{\beta}/{\alpha}$) in the CHO-suspension cells. The expression level was 2-3 times higher than that of the transient expression of eelFSH in attached CHO-K1 cells. The molecular weight of the $rec-eelFSH{\beta}/{\alpha}$ protein was identified to be approximately 34 kDa. The cells expressing eelFSHR-t614 showed an increase in agonist-induced cAMP responsiveness. The maximal cAMP responses of cells expressing eelFSHR-t614 were lower than those of cells expressing eelFSHR-wild type (eelFSHR-WT). The $EC_{50}$ following C-terminal deletion in CHO-K1 cells was approximately 60.4% of that of eelFSHR-WT. The maximal response in eelFSHR-t614 cells was also drastically lower than that of eelFSHR-WT. We also found similar results in PathHunter Parental cells expressing ${\beta}$-arrestin. Thus, these data provide evidence that the truncation of the C-terminal cytoplasmic tail phosphorylation sites in the eelFSHR greatly decreased cAMP responsiveness and maximal response in both CHO-K1 cells and Path-Hunter Parental cells expressing ${\beta}$-arrestin.
Park, Sung-Soo;Shin, Dong-Ho;Kim, Tae-Wha;Lee, Dong-Hoo;Lee, Jung-Hee;Lee, Jung-Dal
Tuberculosis and Respiratory Diseases
/
v.40
no.2
/
pp.185-191
/
1993
Background: Intercellular adhesion molecule-1(ICAM-1) is a 90 kD surface glycoprotein, associated with ${\alpha}_L{\beta}_2$ and ${\alpha}_M{\beta}_2$ subunit of integrins, that serve as cell-cell and cell-substratum adhesion molecules and help regulate cellular morphology, differentiation, and proliferation. The adhesion molecules likely play important roles in maintaining the normal structure and function of the lung. ICAM-1 system among many cell adhesion molecules is importantly issuing in the pathogenesis of idiopathic pulmonary fibrosis. Methods : By using $IgG_1$ monoclonal antibody for ICAM-1, we investigated immunohistochemically the expression of ICAM-1 in the formalin-fixed, paraffin-embedded tissue sections of the 3 normal cases and 6 pieces of tissues taken 3 cases with idiopathic pulmonary fibrosis. Results : In the 3 normal cases, the expressions of ICAM-1 were not discernible. Up-regulation of the ICAM-1 expression was showed in the interstitial fibroblast cells of alveolar septa in 5 pieces and proliferated alveolar pneumocytes in 1 piece among 6 pieces of tissues taken 3 cases with idiopathic pulmonary fibrosis. Conclusion : It was concluded from these findings that up-regulation of the ICAM-1 expression may be related to pathogenesis of idiopathic pulmonary fibrosis.
Jin, Bo-Ram;Choi, In Young;Hwang, Do Young;Ham, Seong-Ho;An, Hyo-Jin
The Korea Journal of Herbology
/
v.34
no.5
/
pp.21-28
/
2019
Objectives : In present study, we investigated a therapeutic effect and optimum dose of Socheongryong-Tang (SCT) on LPS-induced lung inflammation rats model. Methods : Male Sprague-Dawley rats ($260{\pm}10g$) were divided into 12 groups : Group 1 included the normal rats, and Group 2-12 were administrated LPS by intranasal injection to induce experimental lung inflammation. After 1 day of LPS administration, Group 3-9 were treated with SCT ${\times}1/4$, ${\times}1/2$, ${\times}1$, ${\times}3$, ${\times}6$, ${\times}12$ or ${\times}18$, respectively. Group 10-12 (positive control) were treated with dexamethasone 1 mg/kg or acetylcystein 1.5 mg/kg or diclofenac sodium 0.4 mg/kg, respectively. After sacrifice, bronchoalveolar lavage fluid (BALF) was isolated. The levels of IL-$1{\beta}$, TNF-${\alpha}$, mucin glycoprotein 5AC (MUG5AC) were measured in BALF using enzyme-linked immunosorbent assay (ELISA). Results : LPS injected rats exhibited outstanding lung inflammation manifestations, including increased amount of total cells and neutrophil, and upregulated inflammatory cytokines level in BALF. However, the administration of SCT ${\times}1/4$, ${\times}1/2$ and ${\times}1$ decreased total cells and neutrophil, and suppressed the production of inflammatory cytokines, including $IL-1{\beta}$ and TNF-${\alpha}$, and MUG5AC in BALF. Notably, inhibitory effect of SCT ${\times}1/2$ and ${\times}1$ on the level of TNF-${\alpha}$ was markedly better than that of positive controls, dexamethasone and acetylcystein. Conclusions : Taken together, these results suggest that SCT ${\times}1/2$ and ${\times}1$ has therapeutic effects on LPS-induced lung inflammation rats model.
This study aimed to investigate the signal transduction of phosphorylation sites at the carboxyl (C)-terminal region of equine luteinizing hormone/chorionic gonadotropin receptor (eLH/CGR). The eLH/CGR has a large extracellular domain of glycoprotein hormone receptors within the G protein-coupled receptors. We constructed a mutant (eLH/CGR-t656) of eLH/CGR, in which the C-terminal cytoplasmic tail was truncated at the Phe656 residue, through polymerase chain reaction. The eLH/CGR-t656 removed 14 potential phosphorylation sites in the intracellular C-terminal region. The plasmids were transfected into Chinese hamster ovary (CHO)-K1 and PathHunter Parental cells expressing β-arrestin, and agonist-induced cAMP responsiveness was analyzed. In CHO-K1 cells, those expressing eLH/CGR-t656 were lower than those expressing eLH/CGR wild-type (eLH/CGR-wt). The EC50 of the eLH/CGR-t656 mutant was approximately 72.2% of the expression observed in eLH/CGR-wt. The maximal response in eLH/CGR-t656 also decreased to approximately 43% of that observed in eLH/CGR-wt. However, in PathHunter Parental cells, cAMP activity and maximal response of the eLH/CGR-t656 mutant were approximately 173.5% and 100.8%, respectively, of that of eLH/CGR-wt. These results provide evidence that the signal transduction of C-terminal phosphorylation in eLH/CGR plays a pivotal role in CHO-K1 cells. The cAMP level was recovered in PathHunter Parental cells expressing β-arrestin. We suggest that the signal transduction of the C-terminal region phosphorylation sites is remarkably different depending on the cells expressing β-arrestin in CHO-K1 cells.
To elucidate enzymatic properties of $\alpha$-galactosidases (EC3, 2, 1, 22) from germinated soybean and Aspergillus niger changes in the enzyme activities and oligosaccharide contents during germination of soybean were determined and $\alpha$-galactosidases from germinated soybean and wheat bran culture of Aspergillus niger were purified by ammonium sulfate fractionation, ion exchange chromatography and gel filtration. Their chemical and enzymatic properties were investigated and the results obtained were summarized as follows : 1. $\alpha$-Galactosidase activity of soybean was maximized when it was germinated at $25^{\circ}C$ for 120 hours. And raffinose and stachyose in soybean were decomposed completely after 96 hours and 120 hours of germination, respectively. 2. The highest level of $\alpha$-Galactosidase activity was obtained when Aspergillus niger was grown on wheat bran medium at $30^{\circ}C$ for 96 hours. 3. Soybean $\alpha$-galactosidase was purified by 6.6 fold by ammonium slufate fractionation, ion exchange chromatography on DEAE-Cellulose and Sephadex A-50., and gel filtration on Sephadex G-150. Its specific activity was 825 units/mg protein and the yield was 2.5% of the total activity of crude extracts. 4. Aspergillus niger $\alpha$-galactosidase was purified by 23.7 fold. Its specific activity was 1,229 units/mg protein and the yield was 14% of the total activity of wheat bran culture. 5. The purified $\alpha$-galactosidases of soybean and Aspergillus niger were found to be homogeneous by polyacrylamide gel electrophoresis and by HPLC. 6. Chemical properties of the purified $\alpha$-galactosidases were : 1) The soybean $\alpha$-galactosidase was monomeric and its molecular weight was estimated to be 30,000 by SDS-PAGE whereas the Aspergillus niger $\alpha$-galactosidase was a tetrameric glycoprotein which consisted of identical subunits with molecular weight of 28,000 each.
Park, Eun-Jung;Baek, Ji-Young;Shin, Jae-Il;Lee, Jae-Seung;Kim, Hyon-Suk
Childhood Kidney Diseases
/
v.13
no.2
/
pp.146-152
/
2009
Purpose : We performed this study to evaluate the incidence and clinical significance of antiphospholipid antibodies (aPL Ab) in Korean children with Henoch-$Sch{\ddot{o}}nlein$ purpura (HSP). Methods : The medical records of 62 patients (31 boys and 31 girls) aged $46.0{\pm}3.1$ (1-16) years with a clinical diagnosis of HSP based on the EULAR/PReS criteria were reviewed retrospectively. From the years 2007 to 2009, the sera from children with acute HSP were tested for aPL Ab such as LA, anti-cardiolipin antibody and anti-${\beta}_2$ glycoprotein I antibody. Results : LA was positive in 18 (29%) of the 62 patients with HSP and We divided the patients into the two groups LA positive group (N=18) and LA negative group (N=44). There were no significant differences between the two groups with regard to abdominal pain, arthralgia and renal involvement, but LA positive group had significantly higher C-reactive protein ($4.3{\pm}7.2$ mg/dL vs. $1.3{\pm}1.8$ mg/dL, P=0.035), erythrocyte sedimentation rate ($37.5{\pm}26.2$ mm/hr vs. $25.1{\pm}22.6$ mm/hr, P= 0.039), IgM ($148.1{\pm}48.4$ mg/dL vs. $114.9{\pm}41.5$ mg/dL, P=0.024), C3 ($143.1{\pm}21.9$ mg/dL vs. $129.7{\pm}24.5$ mg/dL, P=0.048) and C4 levels ($30.9{\pm}6.3$ mg/dL vs. $24.9{\pm}7.8$ mg/dL, P=0.002) compared with LA negative group. Conclusion : We found that the incidence of positive aPL Ab tests was relatively higher in Korean children with HSP and the presence of aPL Ab was associated with acute inflammatory process of HSP. These results suggest that the aPL Ab are involved in the pathogenesis of HSP in children.
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