• Bulletin of the Korean Chemical Society
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• 제32권5호
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• pp.1569-1574
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• 2011

The presence of replication-competent adenovirus (RCA) subpopulations in adenoviral vector products raises a variety of safety issues for development of therapies based on gene therapy. To analyze the differing effects of adenoviral vector and RCA in vivo, we examined alterations in amino acids (AAs) using rat plasma following injection of ${\beta}$-galactosidase expressing recombinant adenovirus (designated rAdLacZ) or RCA. Plasma AAs were examined by gas chromatography-mass spectrometry. A total of 16 AAs were positively measured. In the rAdLacZ group compared to the control group, the level of aspartic acid was significantly increased (Student's t-test), while the level of glutamic acid was significantly reduced. Additionally, in the RCA group compared to the control group, the level of four AAs, valine, leucine, and isoleucine as branched-chain amino acids, and proline were significantly increased, whereas the levels of three AAs, glycine, threonine, and glutamic acid were significantly reduced. Altered plasma free AA metabolic patterns in rAdLacZ and RCA groups, compared with the control group, may explain the disturbance of AA metabolism related to viral infection.

• The Korean Journal of Physiology and Pharmacology
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• 제24권1호
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• pp.69-79
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• 2020

Aging is one of the risk factors for the development of cardiovascular diseases. During the progression of cellular senescence, cells enter a state of irreversible growth arrest and display resistance to apoptosis. As a flavonoid, quercetin induces apoptosis in various cells. Accordingly, we investigated the relationship between quercetin-induced apoptosis and the inhibition of cellular senescence, and determined the mechanism of oxidative stress-induced vascular smooth muscle cell (VSMC) senescence. In cultured VSMCs, hydrogen peroxide (H₂O₂) dose-dependently induced senescence, which was associated with increased numbers of senescence-associated β-galactosidase-positive cells, decreased expression of SMP30, and activation of p53-p21 and p16 pathways. Along with senescence, expression of the anti-apoptotic protein Bcl-2 was observed to increase and the levels of proteins related to the apoptosis pathway were observed to decrease. Quercetin induced apoptosis through the activation of AMP-activated protein kinase. This action led to the alleviation of oxidative stress-induced VSMC senescence. Furthermore, the inhibition of AMPK activation with compound C and siRNA inhibited apoptosis and aggravated VSMC senescence by reversing p53-p21 and p16 pathways. These results suggest that senescent VSMCs are resistant to apoptosis and quercetin-induced apoptosis attenuated the oxidative stress-induced senescence through activation of AMPK. Therefore, induction of apoptosis by polyphenols such as quercetin may be worthy of attention for its anti-aging effects.

• 대한의생명과학회지
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• 제23권2호
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• pp.49-56
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• 2017

Fibroblast growth factor (FGF)-2 is one of the most effective growth factors to increase the growth rate of mesenchymal stem cells (MSCs). Previously, we reported that low dose of FGF-2 (1 ng/ml) induced proliferation of bone marrow-derived mesenchymal stem cells (BMSCs) through AKT and ERK activation resulting in reduction of autophagy and senescence, but not at a high dose. In this study, we investigated the effects of high dose FGF-2 (10 ng/ml) on proliferation, autophagy and senescence of BMSCs for long term cultures (i.e., 2 months). FGF-2 increased the growth rate of BMSCs in a dose dependent manner for a short term (3 days), while during long term cultures (2 months), population doubling time was increased and accumulated cell number was lower than control in BMSCs when cultured with 10 ng/ml of FGF-2. 10 ng/ml of FGF-2 induced immediate de-phosphorylation of ERK1/2, expression of LC3-II, and increase of senescence associated ${\beta}$-galactosidase (SA-${\beta}$-Gal, senescence marker) expression. In conclusion, we showed that 10 ng/ml of FGF-2 was inadequate for ex vivo expansion of BMSCs because 10 ng/ml of FGF-2 induced growth retardation via ERK1/2 de-phosphorylation and induction of autophagy and senescence in BMSCs.

• 농업과학연구
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• 제41권4호
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• pp.265-274
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• 2014

Physiological characteristics of two Korean golden kiwifruit cultivars, 'Halla Gold' and 'Haehyang', were compared to determine the storage potential of fruit. The soluble solid levels of the fruit were 8.9 and 6.9 oBrix in 'Halla Gold' and 'Haehyang' at harvest, respectively but increased up to 15.4 in 'Halla Gold' and 17.5 oBrix in 'Haehyang' after 2 months of storage. Major sugars were fructose and glucose, and sucrose content was relatively low regardless of cultivar. The edible quality of 'Haehyang' was better than 'Halla Gold' because of higher amount of sugars. Firmness of the fruits gradually decreased as the increase of storage period in 'Halla Gold' in both flesh and core tissue. Th firmness loss of 'Haehyang' fruit was faster in the first 2 months and then became slow. After 75 days of storage, the firmness of 'Haehyang' fruit was only 5.2% at harvest. Core tissue was soften enough to eat at ripe stage. Wall modifying enzyme activities including xylanase, ${\alpha}$-L-arabinofuranosi-dase and ${\beta}$-galactosidase were consistently higher in 'Haehyang' and the activity of pectate lyase was more increased than 'Halla Gold' after 2 months of storage. Respiration rate of 'Haehyang' was higher than 'Halla Gold' and further increased after 2 months of storage. Weight loss was much higher in 'Haehyang' which showed higher rate of the firmness loss. The storage potential of golden kiwifruit was estimated to be about 2 months for 'Haehyang' and 3 months for 'Halla Gold' when determined on the basis of the fruit firmness.

• 대한본초학회지
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• 제20권4호
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• pp.1-10
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• 2005

Objectives : We investigated the anti-aging effect on skin with the extract of the needles of red pine, Pinus densiflora. Methods : We measured various effects related to skin such as the anti-oxidant effect, the protection against ultraviolet (UV) irradiation, the inhibition of reactive oxygen species (ROS) generation, the induction of heat shock protein 70 (HSP70), the reduction of matrix metalloproteinase-2 (MMP-2) synthesis and senescent cell. Results : The results were as follows : The extract of the needles of red pine (RP) had the potent anti-oxidant effect and the ROS scavenging effect. Also RP preserved the systemic anti-oxidant enzyme system (superoxide dismutase and catalase) from UVB irradiation. RP protected the cell membrane from the damages induced by UVB irradiation. RP induced HSP70, a mediator of resistance to UVB irradiation. RP reduced the synthesis of MMP-2 induced by UVB irradiation. And RP inhibited the amount of senescent-associated (SA) ${\beta}-galactosidase$ staining, as a marker of replicative senescence. Conclusions : The results of our study indicate that the extract of the needles of red pine, Pinus densiflora, has anti-aging effects on skin.

• KSBB Journal
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• 제11권3호
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• pp.317-322
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• 1996

식품 및 화장품의 첨가제로 사용되는 기능성 올리고당인 갈락토올리고당을 생룰로스 가수분해 효소를 이용하여 생산하였다. 여섯가지의 효소 시료 가운데, P Penicillium funiculosum에서 유래한 셀룰로스 가수 분해 효소가 가장 높은 올리고당 생산 수율을 보였다. 올리고당 생산을 위한 최적온도와 pH는 각각 $50^{\circ}C$,와 5.0이었으며 유당농도와 효소농도사at의 최 적비는 lO(w/w) 이었다. 시간에 따른 올리고당 생산 반응의 경향성은 유당분해 효소 반응과 유사하였 으며 초기 유당농도가 증가함에 따라 생성되는 올리 고당의 농도가 증가하였다. 최척반응조건에서 20% (w/v)의 유당으로 부터 최고 23%(w/w)의 올리고 땅을 얻을 수 있었다. 생성된 올라고당의 성분은 고 속액체 크로마토그래프와 박층 크로마토그래프 분석 을 통하여 삼당과 사당으로 이루어져 있음을 확인하였다.

• 생명과학회지
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• 제12권6호
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• pp.803-811
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• 2002

초석잠 헥산 추출물이 식중독균에 대한 천연 항균 활성 가능성을 검토하였다. 초석잠 헥산 추출물의 항균 작용을 알아보기 위하여 공시 균주인 Bacillus cereus, Staphylococcus aureus, E, coli 및 Listeria monocytogenes등에 대하여 disc법으로 실험한 결과 그람음성 및 그람 양성균에 대하여 15~20 mm의 clear zone이 나타났다. 초석잠 헥산 추출물의 MIC는 Bacillus cereus에 대해 250${\mu}g$/$m\ell$ Listeria monocytogenes에 대해 250~500/$m\ell$그리고 Staphylococcus aureua와 Pseudomonoas aeruginosa에 대해 각각 500${\mu}g$/$m\ell$ 나타내었다. 초석잠 추출물을 처리한 전자현미경(TEM)상에서는 처리균주들의 세포막이 파괴되어 세포내용물이 용출된 것을 볼 수 있었다. 초석잠 추출물을 처리한 균주의 세포막 손상의 정도를 알아보기 위하여 균체내 효소인 $\beta$-galactosidase 활성을 측정한 결과 클로로포름보다 세포막을 더 손상시키는 것이 확인되었다. 그리고 초석잠 추출물의 pH 및 열 안정성을 실험한 결과 매우 안정하였다.

• 한국축산식품학회지
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• 제33권5호
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• pp.565-571
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• 2013

An enzyme ${\beta}$-galactosidase or ${\beta}$-galactohydrolase [EC3.2.1.23], commonly called lactase, mediates galacto-oligosaccharide (GOS) synthesis under conditions of high substrate concentrations. Also, lactase hydrolyzes ${\beta}$($1{\rightarrow}4$) lactose into glucose and galactose, the latter is successively transferred to free lactose to make various oligosaccharides via transgalactosylation. GOS is non-digestible to human digestive enzymes and has been used as a functional prebiotics. Among the 24 lactic acid bacteria (LAB) strains used, Lactobacillus paracasei YSM0308 was selected based on its exhibition of the highest ${\beta}$-galactoside hydrolysis activity, and the crude lactase was prepared for examination of reaction conditions to affect the GOS synthesis. Lactase activity was measured with a spectrophotometer using ONPG (o-nitropheyl ${\beta}$-D-galactopyranoside) method. Lactase activity was not detected in the culture supernatant and was mostly present in the cell pellet after centrifugation. Activity of the crude lactase preparation ranges from102 to 1,053 units/mL, with the highest activity determined for L. paracasei YSM0308. Optimal conditions for GOS synthesis are as follows: concentration of whey powder, pH, temperature, and time were 30%, pH 6.5-7.0, $30^{\circ}C$, and 4 h, respectively. The final GOS concentration was 19.41% (w/v) by the crude YSM0308 lactase, which was obtained from strain YSM0308 grown in the 10% (w/v) reconstituted whey-based medium.

• 미생물학회지
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• 제47권1호
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• pp.1-6
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• 2011

암의 유전자치료에서 암세포로의 선택적 유전자전달 매체의 부족은 치료효과의 감소를 야기하는 문제이다. 본 연구에서는 난소암 유전자치료의 효율을 높이기 위한 목적으로 난소암세포로 선택적인 유전자전달을 하도록 개량된 아데노바이러스 벡터를 제조하고, 그 효율성을 난소암세포주를 이용하여 조사하였다. 난소암세포에 과다발현하는 분자인 ErbB receptor를 표적하도록 아데노바이러스 외피단백질 fiber에 ErbB receptor에 대한 ligand인 heregulin으로부터 유래한 펩티드를 부착하였다. 53개의 아미노산으로 구성된 외부 펩티드를 fiber에 부착하였을 때 바이러스 감염에 중요한 기능을 하는 fiber 삼량체 구조 형성에 영향을 미치지 않았다. Fiber를 조작한 개량 아데노바이러스는 야생형 fiber를 가진 1세대 아데노바이러스 벡터에 비해 선택적으로 난소암으로 유전자를 전달하는 비율이 증가하였다. 특히 항암제에 저항성을 가진 난소암세포주 OVCAR3에서 유전자전달 효율이 약 5배 증가되었다. 따라서 난소암의 유전자치료에서 개량된 아데노바이러스로 치료 유전자를 전달하면 치료의 효율성을 향상시킬 수 있을 것이다.

• 한국잠사곤충학회지
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• 제40권1호
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• pp.52-59
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• 1998

For the construction of plasmid and bmNPV sarrying the FRT recognition site for the FLP recombinases, we synthesized the wild type FRT dligonucleotides. The target FRT sequences consist of three 13bp repeated DNA sequences; two repeats in a direct orientation and one inverted relative to the other two. In addition, there is an 8bp spacer region between the repeats which determune the orientation of the FRT recombination site. In order to place the FRT site both in target BmNPV genome and the transfer vector, we constructed a plasmid, FRT site both in the target BmNPv genome and the transfer vector, we constructed a plasmid, pFRT$\beta$-gal, carrying the FRT sites within the cloning sites of pSV vector and a recombinant BmNPV, vFRTPH, carrying the FRT sites at a downstream of polyhedrin promotor, respectively. In order to test the functionality of the FLP/FRT site-specific recombination system, vFRTPH, pFRT$\beta$-gal and pHsFLP DNA were co-transfected into BmN-4 cells. The resulting recombinant virus was designated a vFRT$\beta$2-gal. From construction analysis of the vFRT$\beta$2-gal with PCR technique it was concluded that the entire pFRT$\beta$-gal plasmid with $\beta$-galactosidase gene and origines of replication flanked by two functional hybrid FRT sequences. The efficiency of recombination was 8.7%, which was higher than that(2.2%) of recombination between a conventional transfer vector and the wild type BmNPV.