Glycoproteins isolated from fruit bodies and mycelial cultures of mushrooms exhibit anti-carcinogenic actions in human cancer cells and animal tumor cells by induction of apoptosis. Here, we report that isoflavone-conjugated glycoproteins (designate Gluvone), exhibit strong anti-carcinogenic effects on human breast cancer MCF-7 cells by induction of apoptosis. Gluvone with 9.4 kDa of molecular weight was isolated from submerged-liquid culture of Agaricus blazei mycelia (ABM) in soy flake-containing liquid medium. MCF-7 cells were incubated with various amounts of Gluvone (0~250 μM) for a period of 6 days. Gluvone exhibited anti-proliferative actions in a dose-dependent manner and 62% growth inhibition at 200 μM for 4 days relative to control. Hoechst 33258 staining analysis revealed that Gluvone induced formation of apoptotic bodies. Gluvone was associated with down-regulation of anti-apoptotic Bcl-2 protein expression as well as up-regulation of pro-apoptotic Bax protein expression. Gluvone treatment induced proteolytic activation of caspase-9 and caspase-3 through cytochrome c release from mitochondria to cytosol as well as concomitant degradation of poly (ADP-ribose) polymerase (PARP). In addition, Gluvone induced activation of caspase-8. Taken all together, these results indicate that the anti-proliferative effect of Gluvone is associated with induction of apoptotic cell death through the mitochondrial dysfunction pathway mediated by enhancement of Bax protein expression and suppression of Bcl-2 protein expression.

본 연구에서 당근에서 제조한 CG의 기능성 화장품 소재로서의 응용성을 SCP와 비교 검토하기 위하여 CG와 SCP를 적용한 각각의 기능성 크림을 제조하였다. CG와 SCP를 적용한 각각의 기능성 크림에서의 변색, 변취 및 크리밍 현상이나 응집 등의 관능검사와 pH, 점도 등의 물성 변화는 5℃, 25℃에서 매우 안정하다. 피부에서의 수분손실량 및 수분 함유량의 변화를 측정한 결과에서도 CG의 보습작용이 우수한 것으로 나타났다. 이러한 결과를 토대로 CG를 함유한 크림은 SCP를 함유한 크림과 마찬가지로 피부에 보습 효과가 우수하여 주름개선에도 큰 영향을 줄 것으로 예상된다.

The silkworm-baculovirus expression system has distinct advantages, such as a high yield and safe usage in vertebrates. Here, we report a novel strategy for the large-scale production of a classical swine fever virus (CSFV) envelope glycoprotein E2 in the larvae of a baculovirus-infected silkworm, Bombyx mori. We constructed a recombinant B. mori nucleopolyhedrovirus (BmNPV) that expressed recombinant polyhedra together with the N-terminal 179 amino acids of CSFV E2 (E2ΔC). BmNPV-E2ΔC-infected silkworm larvae expressed native polyhedrin and approximately 44-kDa fusion protein that was detected using both anti-polyhedrin and anti-CSFV E2 antibodies. Electron and confocal microscopy both demonstrated that the recombinant polyhedra contained both the fusion protein and native polyhedrin were morphologically normal and contained CSFV E2ΔC. The CSFV E2ΔC antigen produced in BmNPV-E2ΔC-infected silkworm larvae reached 0.68 mg per ml of hemolymph and 0.53 mg per larva at 6 days post-infection. Six-week-old female BALB/c mice that were immunized with the E2ΔC protein purified from solubilized recombinant polyhedraelicited CSFV E2 antibodies, which indicated that the CSFV E2ΔC protein from recombinant polyhedra was immunogenic. The virus neutralization test showed that the serum from mice that were treated with E2ΔC protein from recombinant polyhedra contained significant levels of virus neutralization activity. These results demonstrate that the present strategy can be used for the large-scale production of CSFV E2 antigen.

Classical swine fever virus (CSFV) envelope glycoprotein E2 is the main target for inducing neutralizing antibodies and protective immunity in swine. Here, we report a novel strategy forthe large-scale production of a CSFV E2 subunit vaccine that demonstrates a high immunogenic capability in the larvae of a baculovirus-infected silkworm, Bombyx mori. We constructed a recombinant B. mori nucleopolyhedrovirus (BmNPV) that expressed recombinant polyhedra together with the N-terminal 179 amino acids of CSFV E2 (CSFV E2ΔC). BmNPV-E2ΔC-infected silkworm larvae expressed an approximately 44-kDa fusion protein that was detected using both anti-polyhedrin and anti-CSFV E2 antibodies. Electron and confocal microscopy both demonstrated that the recombinant polyhedra were morphologically normal and contained CSFV E2ΔC. The CSFV E2ΔC antigen produced in BmNPV-E2ΔC-infected silkworm larvae reached 0.68 mg per ml of hemolymph and 0.53 mg per larva at 6 days post-infection. Mice that were immunized with the granule form of recombinant polyhedra or the soluble form of the fusion protein elicited CSFV E2 antibodies, which indicated that the recombinant polyhedra carrying CSFV E2ΔC were immunogenic. The virus neutralization test showed that the serum from mice that were treated with recombinant polyhedra or the soluble form of the fusion protein contained significant levels of virus neutralization activity. These results demonstrate that the present strategy can be used for the large-scale production of CSFV E2 antigen and that the recombinant polyhedra containing CSFV E2ΔC as a granule antigen can be used as a potential subunit vaccine against CSFV.

Pseudorabies virus (PRV), a member of the Alphaherpesviridae, is the causative agent of Aujeszky’s disease in pigs. Glycoprotein B (gB) of PRV, a major constituent of the viral envelope, consists of 916 amino acids. We continuously combined three gB epitopes, E1 (aa 62-129), E2 (aa 217-282), and E3 (aa 543-737). The DNA fragment containing the PRV gB epitopes was fused with polyhedrin gene in order to generate recombinant baculovirus that expresses the recombinant polyhedra with PRV gB epitopes under the control of the Bombyx mori nucleopolyhedrovirus polyhedrin promoter. Recombinant baculoviruses were injected into fifth-instar B. mori larvae. SDS-PAGE and Western blot analyses revealed that recombinant polyhedra constitute polyhedrin and PRV gB epitopes, and that the recombinant PRV gB epitopes showed cross-reactivity against antiserum of PRV gB produced from pig. To examine the immunogenicity of recombinant PRV gB epitopes, we injected into mice as model animals. ELISA results indicated that antibody production is increased in a similar manner in the injection of recombinant polyhedra with PRV gB epitopes, either injected recombinant polyhedra as a granule form antigen without adjuvant or injected recombinant polyhedrin as a soluble form antigen with adjuvant. Taken together, these data show that PRV gB epitopes were produced as a granule form antigen by fusing recombinant polyhedra in baculovirus-infected silkworm larvae and displayed the immunogenicity in mice, indicating the efficacy of the granule form antigen as a PRV gB vaccine.

The Classical Swine Fever Virus (CSFV) is a member of the Pestivirus genus of the Flaviviridae. The genome of CSFV is a positive single-stranded RNA molecule 12.3 kb and contains a single large open reading frame (ORF). The polyprotein composed of eight nonstructural and four structural proteins (nucleocapsid protein C and three envelope glycoprotein E0, E1 and E2). E2, the most immunogenic of the CSFV glycoproteins, induces a protective immune response in swine. To determine the characteristics of the CSFV, LOM strain, we investigated the nucleotide sequence of the glycoprotein E0, E1 and E2. Comparison of the LOM with the other strains revealed nucleotide sequence identity ranging from 97 to 98%. Expression of the glycoprotein E2 was identified by SDS-PAGE and Western blot analysis using anti-CSFV E2 monoclonal antibodies in Sf21 cells. The expression levels of glycoprotein E2 were observed from day 3 and 5 days maximum. In addition, its expression efficiency by media and cell line was investigated. The result showed that High-Five cells and Grace’s insect media for Sf21 were the best conditions for the expression of the glycoprotein E2.

This study is on the feasibility of use of glycoprotein in various areas such as cosmetics and food etc. by extracting, isolating and refining glycoprotein from carrots, red ginseng extract residue, sesame and pine needles using protease(pepsin) and by analyzing general characteristics and measuring various bioactivities. The results of analysis of nutritional composition showed protein contents of glycoprotein. In the analysis of constitutive amino acids, the ratio of contents of hydroxy proline and glycine, the characteristics of glycoproteins appeared similar and the contents of glutamic acid and aspartic acid appeared higher. As a result of measurement contents of total polyphenol and flavonoid, it showed that glycoprotein had more contents generally, and the effect of bioactivity of glycoprotein appeared higher although different kinds of glycoprotein showed a little DPPH radical and nitrite scavenging ability, total antioxidant capacity by ABTS, ACE inhibitory.

As a herbal supplement, Dioscorea batatas Decne (DBD) presents potent antioxidant activity and diverse health benefits. In the present study, functions of a 30 kDa glycoprotein isolated from DBD (hereafter, DBD glycoprotein) in the regulation of feed efficiency and fecal malodor in mice were explored. DBD glycoprotein produced protective effect against cytotoxicity induced by the ecotoxicological endocrine-disrupting substance bisphenol A in gastrointestinal epithelial HT-29 cells. To investigate its potential roles in the regulation of feed efficiency and fecal malodor, mice were administered an oral injection of DBD glycoprotein for 2 weeks. Compared with the control values, the weight of internal organs (liver, heart, kidney, and spleen) and levels of glutamate pyruvate transaminase, glutamate oxaloacetate transaminase, and lactic dehydrogenase were not significantly changed during DBD glycoprotein administration for 2 weeks. Interestingly, DBD glycoprotein improved feed efficiency and reduced hydrogen sulfide concentration without altering the ammonia level in mouse feces. Collectively, these results indicate that DBD glycoprotein is a functional agent that exerts gastrointestinal protective effects against ecotoxicological substances, improves feed efficiency, and reduces fecal malodor.

Ulmus davidiana Nakai (UDN) has been traditionally used as a herbal medicine in Korea. In the present study, we investigated the anti-ecotoxic potential of a 116 kDa glycoprotein isolated from UDN (UDN glycoprot ein) in regulating fecal malodor and feed efficiency in mice. We found that UDN glycoprotein (200 μg/ml) has an inhibitory effect on the cell death induced by an ecotoxicological endocrine disrupting chemical, bisphenol A, in colon epithelial HT-29 cells. UDN glycoprotein did not show significant differences regarding the weight of ecotoxicity-related organs (liver, heart, kidneys, and spleen) and the levels of serum glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, and lactate dehydrogenase in mice for 2 weeks, compared to the control. Additionally, UDN glycoprotein reduced the levels of hydrogen sulfide and ammonia as markers of fecal malodor in mice. Interestingly, UDN glycoprotein can improve the mouse feed efficiency. In conclusion, our data indicate that anti-ecotoxicological UDN glycoprotein has the ability to increase the feed efficiency and reduce the fecal malodor by maintaining the viability of colonic epithelial cells in mice.

관상동맥 스텐트 혈전증 치료에 있어서 혈소판 당단백 IIb/IIIa 수용체 차단제와 카테터를 이용한 혈전 흡입술을 동시에 시행한 경우의 효과에 대하여 알아보고자 하였다. 2008년 07월부터 2017년 07월까지의 일대학교병원 심혈관센터에서 경피적 관상동맥 스텐트 삽입술 후 관상동맥 조영술에서 스텐트 혈전증이 발생한 환자 267명(64.6±12.1년, 187남자) 그룹 I(혈전용해술과 혈전흡입술 동시 시행한 경우, n=32명), 그룹 II (혈전용해술이나 혈전흡입술 한 가지만 시행한 경우 혹은 둘 다 시행하지 않은 경우, n=235명)로 분류하여 주요심장사건, 사망 발생률, 표적병변 재개통술 그리고 스텐트 혈전 등을 1 년동안 추적 관찰하였다. 두 군 간에 임상 특성에서 연령(그룹I: 60.8±12.9 vs. 그룹II: 65.1±11.9, p=0.603), 남성(그룹I: 75.0% vs. 그룹II: 69. 4%, p=0.681), 좌심실구혈율(그룹I: 58.1±9.0 vs. 그룹II: 59.5±11.9, p=0.127)등 양군 간에 차이는 없었다. 주요 심장사건은 두 군간에 차이를 보이지 않았으나(그룹I: 12.5% vs. 그룹II: 23.8%, p=0.180), 세부적으로 살펴보면 사망발생률(그룹I: 0% vs. 그룹II: 13.2%, p=0.034), 표적병변 재개통술(그룹I: 9.4% vs. 그룹II: 6.4%, p=0.4 61) 그리고 스텐트 혈전증(그룹I: 3.1% vs. 그룹II: 4.7%, p=1000)으로, 사망발생률은 혈소판 당단백 IIb/IIIa 수용체 차단제와 혈전흡입술을 동시에 시행한 군에서 유의하게 낮았다. 결론적으로, 관상동맥 스텐트 혈전증 치료에 있어 혈전용해술과 혈전흡입술을 동시 시행하는 경우에 비교군보다 사망 발생율을 감소시켰다.

Ulmus davidiana Nakai (UDN) has been traditionally used as a herbal medicine to treat inflammatory diseases in Korea. In the present study, we investigated the anti-ecotoxic potential of a 116 kDa glycoprotein isolated from UDN (UDN glycoprotein) in human intestinal epithelial INT-407 cells. We demonstrated that UDN glycoprotein (20 μg/mL) could inhibit the production of lactate dehydrogenase (LDH) induced by toluene, an ecotoxic substance. Additionally, we found that the toluene-induced intestinal cytotoxicity was mediated by the phosphorylation of p38 Mitogen-Activated Protein Kinase (MAPK) via the production of intracellular Reactive Oxygen Species (ROS). The UDN glycoprotein significantly decreased the levels of ROS production and p38 MAPK activation in toluene-stimulated INT-407 cells. Moreover, the UDN glycoprotein inhibits the phosphorylation of nuclear factor-kappa B (NF-κB), which is responsible for the production of LDH, in toluene-stimulated INT-407 cells. Collectively, our data indicate that UDN glycoprotein is a natural antioxidant and a modulator of ecotoxicity signaling pathways in human intestinal epithelial cells.