• Journal of Microbiology and Biotechnology
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• 제34권3호
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• pp.663-672
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• 2024

The use of nanoparticles as a delivery system for a specific antigen could solve many limitations of mucosal vaccine applications, such as low immunogenicity, or antigen protection and stabilization. In this study, we tested the ability of nasally administered chitosan nanoparticles loaded with glycoprotein B of murine cytomegalovirus to induce an immune response in an animal model. The choice of chitosan nanoparticle type was made by in vitro evaluation of sorption efficiency and antigen release. Three types of chitosan nanoparticles were prepared: crosslinked with tripolyphosphate, coated with hyaluronic acid, and in complex with polycaprolactone. The hydrodynamic size of the nanoparticles by dynamic light scattering, zeta potential, Fourier transform infrared spectroscopy, scanning electron microscopy, stability, loading efficiency, and release kinetics with ovalbumin were evaluated. Balb/c mice were immunized intranasally using the three-dose protocol with nanoparticles, gB, and adjuvants Poly(I:C) and CpG ODN. Subsequently, the humoral and cell-mediated antigen-specific immune response was determined. On the basis of the properties of the tested nanoparticles, the cross-linked nanoparticles were considered optimal for further investigation. The results show that nanoparticles with Poly(I:C) and with gB alone raised IgG antibody levels above the negative control. In the case of mucosal IgA, only gB alone weakly induced the production of IgA antibodies compared to saline-immunized mice. The number of activated cells increased slightly in mice immunized with nanoparticles and gB compared to those immunized with gB alone or to negative control. The results demonstrated that chitosan nanoparticles could have potential in the development of mucosal vaccines.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.236-243
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• 2023

혈소판은 1차 및 2차 지혈에서 근본적인 역할을 하는 세포지만 혈소판의 과도한 활성화는 혈전증을 유발할 수 있다. 따라서 혈소판 응집의 적절한 조절은 혈전증 매개 질환을 예방하는 데 중요하다. 최근 곤충소재의 개발이 주목을 받고 있다. 다양한 곤충 자원 중 고영양 기능성 식품원으로는 쌍별귀뚜라미(Gryllus bimaculatus)와 같은 곤충류가 있다. 쌍별귀뚜라미 는 고단백 및 불포화지방산을 함유하고 있으며 2015년 9월 식품의약품안전처로부터 식품원료로 등록되었다. 본 연구에서는 쌍별귀뚜라미 에탄올 추출물(G. bimaculatus extract)이 혈소판 응집, 세포 내 Ca²⁺ 조절, thromboxane A₂ 생산 및 glycoprotein IIb/IIIa (integrin αIIb/β3) 활성화를 억제하는지 여부를 확인하고. 1, 4, 5-triphosphate receptor type I, extracellular signal-regulated kinase, cytosolic phospholipase A₂, mitogen-activated protein kinases p38, vasodilator-stimulated phosphoprotein, phosphatidylinositol-3 kinase, Akt, glycogen synthase kinase-3α/β 및 SYK 같은 신호 분자를 조절할 수 있는지 여부를 조사했다. 우리는 쌍별귀뚜라미 추출물이 혈소판 관련 혈전증 및 심혈관 질환을 예방할 수 있는 잠재적인 치료 약물로 가치가 있음을 규명하였다.

• 한국식품과학회지
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• 제40권6호
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• pp.691-695
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• 2008

본 연구는 콜레스테롤 억제효과가 밝혀진 뽕잎에서 당단백질을 추출하여 과산화지질 라디칼 억제능력 및 생쥐의 혈장 콜레스테롤 수준과 간 해독효소 활성의 개선효과를 평가하였다. In vivo에서 생쥐에게 뽕잎 당단백질을 20 mg/kg의 농도로 섭취시킨 후 콜레스테롤의 수치를 알아보고, 한편, Triton WR-1339를 투여한 생쥐 그룹에서 혈액 및 간 조직을 적출하여 혈장 콜레스테롤의 수준 변화 및 해독효소의 활성을 측정한 결과, 20 mg/kg의 농도로 뽕잎 당단백질을 섭취시킨 생쥐그룹에서 총 콜레스테롤과 LDL 콜레스테롤의 수준이 급격히 감소하였고, HDL의 경우 증가하는 것을 확인할 수 있었다. 또한 Triton WR-1339를 투여한 고지혈증을 유도한 생쥐 그룹에서도 뽕잎 당단백질의 농도에 따른 총 콜레스테롤과 LDL 콜레스테롤의 유의적인 저하를 확인할 수 있었다. 간의 해독효소 중 항산화 기능을 가지고 있는 SOD, CAT 그리고 GPx의 활성은 뽕잎 당단백질의 섭취에 의해 모두 증가하였으나 특히 SOD, CAT의 활성이 크게 나타났고, 그에 비하여 GPx는 그 유의적 차를 느끼기 어려울 정도의 변화 뿐이었다. 일반적으로, 천연물로부터 추출한 당단백질은 항산화 능력을 가지고 있고, 또한 이런 항산화제는 동맥 혈관조직이나 백혈구를 손상시키는 체내에서 발생된 ROS를 제거하고 체내 혈장지단백질 수준에 있어서 반비례적인 상관관계가 있으며, 결과적으로 혈장 콜레스테롤 수준을 억제할 수 있다고 추론할 수 있다(4,25,26). 따라서 이러한 결과에 미루어볼 때, 뽕잎 당단백질 역시 간 내해독효소의 활성을 증가시킴으로써 체내의 ROS 수준을 감소시키고, 콜레스테롤의 수준을 낮추었으므로 뽕잎 당단백질의 역할이 다른 천연물 유래의 당단백질과 마찬가지로 지단백질과 반비례적인 상관관계가 있는 항산화제 역할을 수행할 수 있다고 사료된다. 앞으로 HMG-CoA reductase에 의한 콜레스테롤 생성과 그게 관련된 유전자 발현 및 그 기전을 분자생화학적인 수준에서 보충적인 연구가 더 수행되어야 할 것이다.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1990년도 Proceedings of International Symposium on Korean Ginseng, 1990, Seoul, Korea
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• pp.79-85
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• 1990

The aqueous extract of fresh ginseng (Panax ginseng C.A. Meyer) contains a macromolecular fraction that showed mitogenic and comitogenic activities in human peripheral blood Iymphocytes. Purification of the crude extract by size (ultrafiltration, Sephadex G-200) and charge (DEAE-cellulose. DEAE-Sepharose) yielded a semi-purified fraction (DS-3). This fraction contains at least three subgroups of anionic macromolecules with apparent molecular weight greater than 600 kilodaltons. It is a glycoprotein with a large amount of glucuronic acid. It acts as a mitogen in both T and B cells of human peripheral blood lymphocytes. It could also potentiate the mitogenic action of Concanavalin A in Iymphocyte T cells. Such potentiation is not due to increased binding of Concanavalin A to the cell surface. Its mitogenic and co-mitogenic effects do depend on the presence of extracellular Ca2+.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.193-199
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• 2010

We report here the production of transgenic chickens that can regulate human erythropoietin (hEPO) gene expression. The glycoprotein hormone hEPO is an essential for viability and growth of the erythrocytic progenitors. Retrovirus vector system used in this study has two features including tetracycline-controllable promoter and woodchuck hepatitis virus posttranscriptional regulator element (WPRE). The former is for to reduce the possibility of physiological disturbance due to constitutional and unregulated expression of hEPO gene in the transgenic chicken. The latter is for maximum expression of the foreign gene when we turn-on the gene expression. A replication-defective Moloney murine leukemia virus (MoMLV)-based vectors packaged with vesicular stomatitis virus G glycoprotein (VSV-G) was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 325 injected eggs, 28 chicks hatched after 21 days of incubation and 16 hatched chicks were found to express the hEPO gene delivered by the vector. The biological activity of the recombinant hEPO in transgenic chicken serum was comparable to its commercially available counterpart. The recombinant hEPO in transgenic chicken serum had N- and O-linked carbohydrate simillar to that produced from in vitro cultured cells transformed with hEPO gene.

• 한국어병학회지
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• 제16권1호
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• pp.1-12
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• 2003

RT-PCR method was applied to detect and clone the nucleocapsid protein (N) gene and glycoprotein (G) gene for sequencing 5 Korean VHSV isolates from cultured olive flounder, Paralichthys olivaceus. Phylogenetic analysis was performed to investigate their relationship with the VHSV strains described previously and isolated from different geographical area. Generally, VHSV strains were separated phylogenetically according to the major geographical area of isolation: Genogroup I (American type), Genogroup Il (British Isles) and Genogroup ill (European type). This study revealed that all 5 Korean VHSV isolates were belonged to Genogroup I and closely related to Japanese Obama25 type.

• The Korean Journal of Physiology and Pharmacology
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• 제2권1호
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• pp.119-131
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• 1998

It has been reported that the glycoprotein extracted from Aloe has strong anti-inflammatory response. However, there has been no research report yet about the effect of Aloe on allergic hypersensitivity reactivity. By using guinea pig lung mast cells, this study aimed to examine the effects of Aloe glycoprotein (NY945) on the mediator releases caused by mast cell activation, and also aimed to assess the effects of NY945 on the mechanism of mediator releases in the mast cell activation. We partially purified mast cell from guinea pig lung tissues by using the enzyme digestion, the rough and the discontinuous density percoll gradient method. Mast cells were sensitized with IgG1 (anti-OA) and challenged with ovalbumin. Histamine was assayed by fluorometric analyzer, leukotrienes by radioimmunoassay. The phospholipase D activity was assessed by the production of labeled phosphatidylalcohol. The amount of mass 1, 2-diacylglycerol (DAG) was measured by the $[^3H]DAG$ produced when prelabeled with $[^3H]myristic$ acid. The phospholipid methylation was assessed by measuring the incorporation of the $[^3H]methyl$ moiety into phospholipids of cellular membranes. Pretreatment of NY945 (10 ${\mu}g$) significantly decreased histamine and leukotrienes releases during mast cell activation. The decrease of histamine release was stronger than that of leukotriene during mast cell activation. The phospholipase D activity increased by the mast cell activation was decreased by the dose-dependent manner in the pretreatment of NY945. The amount of DAG produced by PLC activity was decreased by NY945 pretreatment. The amount of mass 1, 2-diacylglycerol produced by activation of mast cells was decreased in the pretreatment of NY945. NY945 pretreatment strongly inhibited the incorporation of the $[^3H]methyl$ moiety into phospholipids. The data suggest that NY945 purified from Aloe inhibits in part an increase of 1, 2-diacylglycerol which is produced by activating mast cells with antigen-antibody reactions, which is mediated via phosphatidylcholine-phospholipase D and phosphatidylinositol-phospholipase C systems, and then followed by the inhibition of histamine release. Furthermore, NY945 reduces the production of phosphatidylcholine by inhibiting the methyltransferase I and II, which decreases the conversion of phosphatidylcholine into arachidonic acid and inhibits the production of leukotrienes.

• IMMUNE NETWORK
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• 제2권4호
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• pp.227-232
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• 2002

Background: Systemic lupus erythematosus (SLE) is a complex autoimmune disease characterized by diverse clinical manifestations and autoantibody production, which is known to be strongly influenced by genetic factors. Previous studies have revealed the associations of SLE with HLA class II alleles and antiphospholipid antibody system (anticardiolipin antibody (aCL) and anti-${\beta}_2$ glycoprotein I antibody (anti-${\beta}_2$ GPI)). Therefore, we studied the associations of HLA class II alleles with SLE and antiphospholipid antibody system. Methods: The genotyping for HLA-DRB1 and DQB1 alleles were performed in 61 SLE patients and 100 controls by the polymerase chain reaction (PCR)-sequence specific oligonucleotide probe method. ELISA tests for aCL and anti-${\beta}_2$ GPI were performed in 39 of the 61 SLE patients. The results were evaluated statistically by Chi-square test. Results: The frequencies of the HLA-$DRB1^*15$ and $DQB1^*06$ in SLE patients were significantly higher than those in controls. HLA-$DRB1^*12$ was significantly lower in SLE patients than controls. Nine of 39 patients were positive for aCL (IgG) and three were positive for aCL (IgM). One of 39 patients were positive for anti-${\beta}_2$ GPI (IgG) and none of them positive for anti-${\beta}_2$ GPI (IgM). Association of aCL with HLA class II alleles was not observed in our study. Conclusion: According to our results, it was found that HLA-$DRB1^*15$ and $DQB1^*06$ were associated with genetic susceptiblility and $DRB1^*12$ was associated with resistance to SLE in Korean population. No Association of aCL with HLA class II alleles was observed and the positive rate for anti-${\beta}_2$ GPI was very low.

• Journal of Applied Biological Chemistry
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• 제57권2호
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• pp.179-182
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• 2014

${\alpha}$-glucosidase 활성을 저해하는 초두구 메탄올 추출물의 항바이러스 활성을 연구하였다. Newcastle disease virus (NDV) 감염된 baby hamster kidney (BHK) 세포에서 Syncytium (합포체) 형성은 세포막 표면으로의 수송된 바이러스 당단백질 hemagglutinin-neuramidase (HN)에 의해 일어난다. 초두구 추출물$IC_{50}$ $25{\mu}g/mL$) 처리된 세포에서 바이러스 당단백질의 수송과 함께 합포체 형성이 저해되었다. 또한 $IC_{50}$ 농도에서는 세포 내에서 당단백질 생합성은 저해되지 않으며 당단백질의 수송을 저해하는 것으로 추론된다.

• Journal of Pharmaceutical Investigation
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• 제37권5호
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• pp.297-303
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• 2007

The purposes of this study were to clarify the involvement of P-glycoprotein (P-gp) in the efflux of levosulpiride in knockout mice that lack the mdr1a1b gene and to evaluate the relationship between the genetic polymorphisms in MDR1 gene (exon 21) and levosulpiride disposition in healthy Korean subjects. After oral administration ($10\;{\mu}g/g$) of levosulpiride to mdr1a/1b(-/-) and wild-type mice, plasma and brain samples were obtained at 45 min. We also investigated the genotype for MDR1 (exon 21) gene in humans using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. A single oral dose of 25 mg levosulpiride was administered to 58 healthy subjects, who were based on the MDR1 genotype for the G2677T SNP. Blood samples were taken up to 36 hr after dosing. The concentrations of levosulpiride in mouse plasma and brain were statistically significant difference between the two animal groups (P<0.05). In addition, the average brain-to-plasma concentration ratio (Kp) of levosulpiride was 3.4-fold (P<0.01) higher in the mdr1a/1b(-/-) mice compared with the wild-type mice. We also found that the values of $AUC_{0-{\infty}$, partial AUC ($AUC_{0-4h}$) and $C_{max}$ were significantly different between homozygous 2677TT subjects and the subjects with at least one wild-type allele (GG and GT subjects, P=0.012 for $AUC_{0-{\infty}$; P=0.008 for $AUC_{0-4h}$; P=0.038 for $C_{max}$). The results confirm that levosulpiride is a P-gp substrate in vivo, and clearly demonstrate the effect of SNP 2677G>T in exon 21 of the MDR1 gene on levosulpiride disposition.