• Journal of Applied Biological Chemistry
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• 제64권3호
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• pp.309-316
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• 2021

Trifolin (kaempferol 3-O-galactoside)는 플라보놀 그룹에 속하는 물질로 아토피, 항균, 폐암에 효과가 있는 것으로 알려져 있다. Trifolin은 다양한 식물에서 추출하여 사용하고 있지만 추출 과정이 복잡하고, 수율이 낮으며, 추출을 위한 바이오매스를 얻는데 계절적 어려움이 있다. 생물전환은 저렴한 화합물에서 고부가가치 화학물질을 생산할 수 있는 대체 수단으로 이용된다. 본 연구에서는 naringenin으로부터 trifolin을 생합성하기 위해 3개의 유전자(PeFLS 및 OsUGE-PhUGT)를 각각의 대장균에 도입한 BL-FLS균주와 BL-UGTE균주를 이용하여 공조배양시스템을 개발하였다. Naringenin으로부터 trifolin을 생합성하기 위해 세포의 밀도, 생물전환 온도, 재조합 단백질 유도의 적정 IPTG농도 및 시간, 기질 공급 농도 등의 최적화를 실시하였다. 최적화된 공동 배양 발효 시스템을 통해 67.3 mg/L의 trifolin을 성공적으로 생합성 하였다.

• 대한방사성의약품학회지
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• 제5권1호
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• pp.54-60
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• 2019

Sentinel lymph node (SLN) imaging plays an important role in surgery of patients with breast cancer and melanoma. In this study, avidin (Av), a tetrameric protein glycosylated with mannose and N-acetylglucosamine molecules, was labeled with $^{64}Cu$ and then evaluated for LN imaging. $^{64}Cu$-Labeled $NeutrAvidin^{TM}$ (NAv), a non-glycosylated form of Av, was used for comparison. 1,4,7,10-Tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA)-conjugated Av and NAv were prepared from the corresponding proteins and DOTA-NHS ester, which were then labeled with copper-64 and purified using PD-10 columns. The numbers of DOTA molecules conjugated to Av and NAv were 4.9 and 3.3, respectively. [$^{64}Cu$]Cu-DOTA-conjugated Av and NAv were prepared in 93% and 73% radiochemical yields, respectively. In vitro serum stability study showed that copper-64 remained stable on all radiotracers for 24 h (>97%). MicroPET/CT images showed that high radioactivity was accumulated in LNs within 15 min after footpad-injection of radiotracers. Tissue distribution data of mice demonstrated significantly higher uptake in the popliteal (PO) LN than lumbar (LU) LN for $^{64}Cu$-labeled Av (relative % ID/g excluding the injection sites: 66.2% and 26.0%, respectively) compared with those of $^{64}Cu$-labeled NAv (43.0% and 49.2%, respectively). The results of this study suggest that mannose molecules on Av enabled the radiotracer to retain in the first LN after mouse footpad-injection.

• Toxicological Research
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• 제35권2호
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• pp.131-136
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• 2019

Ethylhexyl dimethyl para-aminobenzoic acid (PABA) is an oily yellow liquid derivative of water-soluble PABA commonly used in sunscreen. Ethylhexyl dimethyl PABA is widely used as an ingredient in many cosmetics at an average concentration of 1.25% (0.5-2.0%) in Korea. Previous studies, including those involving animals, have demonstrated that ethylhexyl dimethyl PABA is toxic to the following four organs: testis, epididymis, spleen, and liver. In addition, experiments using human keratinocytes found that ethylhexyl dimethyl PABA inhibits cell growth and DNA synthesis at low concentrations, and halted the cell cycle of MM96L cells (human melanoma cell line) at the G1 phase. Despite limited clinical data in humans, many studies have confirmed increased mutagenicity of ethylhexyl dimethyl PABA following exposure to sunlight, which suggests that this molecule is likely to contribute to onset of sun-induced cancer despite protecting the skin through absorption of UVB. For risk assessment, the no observed adverse effect level (NOAEL) chosen was 100 mg/kg bw/day in a 4 weeks oral toxicity study. Systemic exposure dosage (SED) was 0.588 mg/kg bw/day for maximum use of ethylhexyl dimethyl PABA in cosmetics. Based on the risk assessment and exposure scenarios conducted in this study, the margin of safety (MOS) was calculated to be 180.18 for a sunscreen containing 8% ethylhexyl dimethyl PABA, which is the maximum level allowed by the relevant domestic authorities.

• Journal of Microbiology and Biotechnology
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• 제31권7호
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• pp.990-998
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• 2021

Melanin is a natural skin pigment produced by specialized cells called melanocytes via a multistage biochemical pathway known as melanogenesis, involving the oxidation and polymerization of tyrosine. Melanogenesis is initiated upon exposure to ultraviolet (UV) radiation, causing the skin to darken, which protects skin cells from UVB radiation damage. However, the abnormal accumulation of melanin may lead to the development of certain skin diseases, including skin cancer. In this study, the antioxidant and antimelanogenic activities of the cell-free supernatant (CFS) of twenty strains were evaluated. Based on the results of 60% 2,2-diphenyl-1-picrylhydrazyl scavenging activity, 21% 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) scavenging capacity, and a 50% ascorbic acid equivalent ferric reducing antioxidant power value, Limosilactobacillus fermentum JNU532 was selected as the strain with the highest antioxidant potential. No cytotoxicity was observed in cells treated with the CFS of L. fermentum JNU532. Tyrosinase activity was reduced by 16.7% in CFS-treated B16F10 cells (but not in the cell-free system), with >23.2% reduction in melanin content upon treatment with the L. fermentum JNU532-derived CFS. The inhibitory effect of the L. fermentum JNU532-derived CFS on B16F10 cell melanogenesis pathways was investigated using quantitative reverse transcription polymerase chain reaction and western blotting. The inhibitory effects of the L. fermentum JNU532-derived CFS were mediated by inhibiting the transcription of TYR, TRP-1, TRP-2, and MITF and the protein expression of TYR, TRP-1, TRP-2, and MITF. Therefore, L. fermentum JNU532 may be considered a potentially useful, natural depigmentation agent.

• Biomolecules & Therapeutics
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• 제30권2호
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• pp.162-169
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• 2022

We utilized Fas21, a resveratrol analog, to modulate the function of hepatic stellate cells (HSCs) and liver sinusoidal endothelial cells (LSECs) during the angiogenic phase of murine liver metastasis by B16 melanoma and 51b colorectal carcinoma. Preangiogenic micrometastases were treated with Fas21 (1 mg/kg/day) or vehicle during the development of intra-angiogenic tracts. Mice treated with Fas21 showed reduced liver tumor foci in both liver metastasis models. Micrometastases were classified immunohistochemically, as well as according to their position coordinates and connection to local microvasculature. The volume of liver occupied by sinusoidal-type foci, containing infiltrating angiogenic capillaries, decreased by ~50% in Fas21-treated mice compared to vehicle-treated ones in both tumor metastasis models. The volume of portal foci, containing peripheral neoangiogenesis within a discontinuous layer of myofibroblasts, was similar in all experimental groups in both tumor metastasis models, but displayed enhanced necrotic central areas devoid of angiogenesis following Fas21 treatment. As a result, sinusoidal tumors from mice treated with Fas21 showed a 50% reduction in desmin(+)/asma(+) HSCs and CD31(+) vessel density, and a 45% reduction in intrametastatic VEGF mRNA compared with sinusoidal tumors from vehicle-treated mice. Necrotic portal metastases increased 2-4-fold in treated mice. In vitro, Fas21 reduced VEGF secretion by HSCs and 51b cells dose-dependently. Additionally, HSCs migration in response to tumor soluble factors was dose-dependently diminished by Fas21, as was LSEC migration in response to HSCs and tumor soluble factors. Resveratrol analog Fas21 inhibits the proangiogenic response of HSCs and LSECs during the development of murine liver metastasis.

• IMMUNE NETWORK
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• 제3권1호
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• pp.53-60
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• 2003

Background: The role of macrophages in tumor angiogenesis is known to be the production of angiogenic cytokines and growth factors including TNF-${\alpha}$. Recently, macrophage also can produce the INF-${\gamma}$ that is being studied to be involved in angiogenic inhibition. Thus, the importance of macrophages in tumor angiogenesis is might being an angiogenic switch. Thus, the hypothesis tested here is that TNF-${\alpha}$ can modulate the INF-${\gamma}$ production in the macrophages from tumor environment as a part of tumor angiogenic switch. Methods: Macrophages in tumor environment were obtained from the peritoneal cavity of C57BL/6 mice injected with B16F10 melanoma cell line for 6 or 11 days. $Mac1^+$-macrophages were purified using magnetic bead ($MACs^{TM}$; Milteny Biotech, Germany) and cultured with various concentrations of TNF-${\alpha}$ for various time points at $37^{\circ}C$. The supernatants were analyzed for IFN-${\gamma}$ or VEGF by ELISA kit (Endogen, Woburn, MA). Results: Residential macrophages from the peritoneal cavity did not respond to LPS or TNF-${\alpha}$ to produce INF-${\gamma}$. However, the cells from tumor environment produced IFN-${\gamma}$ as well as VEGF and upregulated by the addition of LPS or TNF-${\alpha}$. RT-PCR analysis revealed the external TNF-${\alpha}$-induced IFN-${\gamma}$ gene expression in the macrophages from tumor environment. Conclusion: The overall data suggest that the macrophages in tumor environment might have an important role not only in angiogenic signal but also in anti-angiogenic signal by producing related cytokines. And TNF-${\alpha}$ might be a key cytokine in tumor angiogenic switch.

• 원예과학기술지
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• 제34권2호
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• pp.331-341
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• 2016

본 연구는 기능성 지표물질 확인을 위한 동양란 심비디움(Cymbidium) 향기 성분 비교 분석을 목적으로 하며, GC/MS 방법을 사용하여 한국춘란(Cymbidium goerigii L.), 중국춘란(C. forrestii R.)과 일경구화(C. faberi R.)의 향기성분을 비교 분석하였다. 분석은 한국춘란으로 '민춘란', '주금화', 중국춘란으로 '취개', '송매', '용자', 일경구화로 '최매', '남양매', '화자'등을 사용하였다. GC/MS 방법을 통해 검출된 성분들을 peak area(%)값 분석을 사용하여 3%이상의 주요 향기성분으로 분류하였다. 향기 성분 분석 결과 한국춘란에서는 유방암, 자궁경부암, 교모세포종암에 세포독성 기능성을 가진 ${\alpha}$-bergamotene과, 인체 간암 세포(HepG2)의 사멸과 성장 억제, 항진균제 및 바베스열원충, 충치균에 대한 억제 기능성을 가진 nerolidol이 가장 많았다. 중국 춘란에서는 nerolidol과 악성 흑색종 세포(B16-F10), 인체 간암 세포와 백혈병 세포(HL-60, K562)의 사멸과 억제 등의 기능성을 가진 ${\beta}$-bisabolene이 가장 많았다. 일경구화에서는 교모세포종(SF-767)의 억제와 간암세포주(BEL-7402)에 소염작용 억제 효과를 가진 ${\alpha}$-pinene, 위장 보호 효과 기능을 가진 1,8-cineole과 페로몬의 기능을 가진 1,3,7-octatriene이 가장 많았다. 이상에서 동양란의 주요 향기 성분으로 밝혀진 물질들은 인간에게 유익한 다양한 기능성을 갖고 있는 것으로 확인되었다.

• 대한한의학방제학회지
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• 제25권2호
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• pp.223-251
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• 2017

Objectives : A lot of experiment results of Yugmijihwangtang(YM) are reported in various kinds of journals. Many of them report on the new effects that are not recorded in the traditional medical texts. So it is necessary to take it into consideration that newly reported effects could be of help to clinical practice, because this process of comparison of Donguibogam and scientific experiment results will have basis to lead into the evidence based medicine. Methods : We compared the effects of in Donguibogam and the experiment results of YM. Results : The effects of YM in Donguibogam are to replenish essence and marrow, and to treat red wen, fatigue, treat hypouresis, urinary sediment, urinary urgency, hematuria, hydrocephalus, speech and movement retardation, yin-deficiency, diabetes mellitus, nonalcoholic fatty liver, melanoma, disability to see near and far sight, tinnitus, hearing loss, alopecia, angiogenesis, cough, cough at night, trachyphonia, and, infantile convulsion. The experiment results of YM since 2000 in both Korea and China are to inhibit atopic dermatitis, renal interstitial fibrosis, anti-oxidant, emphysema, stress, glomerulosclerosis, diabetic nephropathy, chronic glomerulonephritis, hemorrhage, plantar sweating, dermal aging, kidney aging, bone loss, breast cancer, pathological myocardial cell, primary liver cancer, thrombosis, osteoporosis, intrauterine growth retardation, chronic renal failure, IgA nepropathy, slow cerebral development, and hippocampal tissue lesions on the one hand, and to help bone formation, renin-angiotensin- aldosterone system, cerebral recovery, cognitive function and expression, osteoblast proliferation and differentiation, learning and memory, cold-tolerance and oxygen deficit-tolerance and anti-fatigue, endometrial formation, humoral and cell-mediated immunity, immune regulation effect, Hypothalamus-Pituitary-Ovary Axis, and spermatogenesis, on the other hand. Conclusion : When we compared the effects of YM with the experiment results of YM, there existed a considerable gap between them. So, from now on, it is expected that a great effort and consideration are needed to solve these gaps from an academic and clinical point of view.

• Journal of Animal Science and Technology
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• 제44권4호
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• pp.427-442
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• 2002

이상의 연구 결과들을 요약해보면 CLA는 생체내외에서 다양한 생리활성 효과를 가지는 것을 알 수 있다. CLA의 주요 효과는 유방암, 췌장암, 피부암 및 대장암을 억제하는 효과, 동물체내에서의 면역력증강 효과, 동맥경화증에 대한 효과, 지방축적 억제효과, 체내 지방분해 촉진효과 및 식육에서의 항산화 효과와 육색안정 효과 등으로 요약된다. 그러므로 CLA는 건강보조식품으로 혹은 축산식품을 비롯한 다양한 식품의 기능성 첨가제로써의 충분한 가능성을 가지고 있는 것으로 사료된다. 그러나 이러한 CLA의 효과는 실험에 따라 다소간의 차이를 나타내고 또한 CLA의 생리활성 기작에 대한 정확한 이해가 부족한 실정이다. 따라서 향후 CLA와 관련하여 다음과 같은 연구가 필요할 것으로 사료된다. 1) 새로운 CLA 이성체의 발견 및 CLA 합성방법에 대한 연구 2) CLA가 가지는 각각의 생리활성 효과에 대한 정확한 기작을 밝히는 연구 3) CLA 이성체간의 생리활성 효과 차이에 대한 연구 4) CLA의 생리활성 효과를 극대화 할 수 있는 이성체간의 비율에 대한 연구 5) 기능성 식품첨가제로써의 CLA를 효과적으로 이용하는 방법 연구 6) CLA를 식용가축 및 축산물에 효과적으로 축적시키는 방법 연구 7) CLA를 이용한 육제품 및 유제품 개발 연구 8) 사람에 있어 CLA 효과에 대한 임상 연구.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제27권6호
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• pp.483-490
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• 2001

There were many controversies in the cause and progress of tumorigenesis. Recently, studies on the mutation of genes related to the tumor have extensively been performed due to development of molecular biology. Structural and morphological changes of chromosomes, which are related to the abnormal activation of oncogenes or inactivation of tumor suppression genes, transform the normal cells into the tumor cells. p53 and Rb are well known tumor suppressor genes, while oncogenes include c-myc, bcl-2 and ras, etc. When exposed to cell damaging agents, p53 inhibits cell growth by inducing transcription of p21. Especially p73, which is homo-logy of p53, frequently deleted in melanoma, neuroblastoma, colon cancer, and breast cancer, when over produced, p73 activates the transcription of p21, bax-1 and inhibits cell growth by inducing apoptosis. For study on mRNA expression of p21 and p73, normal oral keratinocytes, and cell lines of primary and metastatic oral squamous cell carcinoma were cultured and then electrophoresis and RT-PCR(reverse transcription-polymerase chain reaction) were performed. 1. The mRNA of p21 and p73 in normal oral keratinocyte expressed lower than that of primary squamous cell carcinoma. 2. The mRNA of p21 in metastatic oral squamous carcinoma cell lines was expressed as various patterns compared with that of normal oral keratinocyte. 3. In the metastatic oral squamous cell lines, the mRNA of HN8 expressed higher than that of HN12 or HN19. 4. The mRNA of p73 in primary oral squamous cell lines expressed 4-5 times higher than that of normal keratinocyte. 5. In metastatic oral squamous cell lines, there was no significant expression of p73 mRNA compared with that of normal oral keratinocyte. From the results obtained in this study, mRNA expression of p73 in primary oral squamous cell lines was remarkable, while mRNA expression of p21 and p73 in metastatic oral squamous cell lines were statistically insignificant.