Background : As a part of ongoing research to elucidate and characterize anti-inflammatory nutraceuticals, six kinds of plant extracts (aerial part of Nepeta cataria, leaves of Lonicera maackii, leaves of Platycarya strobilacea, flower of Fagopyrum dibotrys, flowers and fruits of Solanum nigrum, stem of Physostegia virginiana) were tested for their ability to suppress inflammation. The anti-inflammatory has been studied in lipopolysaccharide (LPS)-stimulated RAW264.7 cells which cells synthesized nitric oxide (NO) from L-arginine by nitric oxide synthase (NOS). In this study, NO synthesis inhibitory activity of six kinds of plant extracts on LPS-stimulated RAW 264.7 mouse macrophages was evaluated. Methods and Results : Six kinds of plant extracts were parceled out from RDA (Rural Development Administration). RAW 264.7 cells (1.5×105 cells/well) were seeded onto 96-well plates with DMEM media containing 10% FBS and 1% antibiotics. The cells were pretreated with the extracts and LPS-stimulated cells for 24 h. Cellular NO production was stimulated by adding 1 μg/mL of LPS. After incubation, Griess reagent was used to determine NO production. Absorbance was measured at 520 nm by microplate reader. NO synthesis inhibitory activity potential of these extracts was evaluated by assessing NO production by LPS-stimulated RAW 264.7 cells in the presence. As a result, inhibition rate of NO production was about 40% of L. maackii, 33% of F. dibotrys, 23% of P. strobilacea and 17% of P. virginiana. Meanwhile, there was no significant results in aerial part of N. cataria and flowers and fruits of S. nigrum. Conclusion : From the above results, we be able to confirm that leaves of L. maackii and flower of F. dibotrys appeared dose-dependent NO synthesis inhibitory activity and leaves of P. strobilacea appeared NO synthesis inhibitory activity in low-concentration. As screening NO synthesis inhibition of six extracts, they may be a good candidate for delaying the progression of human inflammatory diseases and warrants further studies.

Background : Ginseng has been commonly used as a traditional oriental medicine for its wide spectrum of medicinal properties, including anti-inflammatory, antitumorigenic, adaptogenic and anti-aging properties. 20(S)-Protopanaxadiol (PPD), a intestinal metabolite of ginsenosides, is one of the active ingredients in ginseng. In this study, we have found inflammation-related genes regulated by 20(S)-PPD in mouse bone marrow-derived macrophage (BMDM) to elucidate the role of 20(S)-PPD in inflammatory signaling pathways. Methods and Results : We examined cell viability of BMDM cells after treatment of 20(S)-PPD and found that 20(S)-PPD has no cytotoxicity up to 10 uM. BMDM cells treated with none or 10uM 20(S)-PPD were used for RNA extraction and microarray analysis. It was found that 2 inflammation-related genes are upregulated and 4 genes are downregulated by 20(S)-PPD. Conclusion : These results can give clues to elucidate the role of 20(S)-PPD in inflammatory signaling pathways.

Background : Recently, ginseng (Panax ginseng C.A. Meyer.) berry has been used as a health-promoting supplements. Also, Mulberries (Morus alba L.) fruit have been used in traditional herbal medicine to treat and prevent diabetes. In this study, we measured the cytotoxicity after fermentation of the extracts in Panax Ginseng Berry and Mulberry Fruit. Methods and Results : The extracts were prepared by decoction for 3 hours in distilled water (100 g/L). The dried extract was then dissolved in phosphate-buffered saline (PBS) in preparation for use. Cell viability was examined by an MTT assay. RAW 264.7 cells were seeded at 1 × 104/mL densities in 96-well plates. Each grouping had a non-treated group as the control. The extracts were added to each well and incubated for 24 hours at 37°C and 5% CO2. The MTT solutions (5 mg/mL) were added to each well, and the cells were cultured for another 2 hours. The supernatant was then discarded, and 100 μL of dimethyl sulfoxide was added to each well. The optical density was read at 540 nm. Conclusion : Probiotics and prebiotics modulate the composition of human and domestic animal gut microbiota. The beneficial effects may result from suppression of harmful microorganisms or stimulation of organisms which contribute in a positive way to the nutrition and health of human and domestic animal. Recently, fermentation using microorganisms for the production of more effective compounds has been extensively studied. In particular, the novel pharmacological effects of a new compound generated by fermentation have been reported. Some previous studies have demonstrated that Fermented herbal medicine extract showed better bioactivity than normal herbal Plants extract when used at the same concentration.

Background : The objective of this study was to Amomum tsao-ko Crevost et Lemarié, (Zingiberaceae) is widely distributed among several countries in South Asia and Southeast Asia. It is a well known spice in Asia, produces a nice refreshing effect in the mouth. Additionally, it's dried fruit is used in Traditional Chinese Medicine (TCM) for cardiac diseases, edema, eye trouble, skin, itch and impotence. The objective of this study was evaluated the inhibitory activity on adipogenesis in 3T3-L1 cells from A. tsao-ko. Methods and Results : The fruits of A. tsao-ko were extracted with 80% EtOH two times at room temperature. The EtOH extract was suspended in distilled water and partitioned with solvent to give CH2Cl2, EtOAc and n-BuOH. The CH2Cl2 was suspended in n-hexane and partitioned with solvent to give 50%, 70% and 90% MeOH. The purification of each fraction by column chromatography separation and HPLC analysis. Consequently, several constituents were isolated five known compounds. The identification and structural elucidation of compounds were established by using NMR (1D and 2D) and mass spectrometry. Conclusion : These compounds were identified as fluorenone (1), phenanthrene (2), anthracene (3), methyl linolenate (4), 1,2-benzenediol (5). All isolates were tested for their inhibitory activities on adipogenesis in 3T3-L1 cells.

Background : Ganoderma lucidum is a non-toxic, medicinal mushroom, which is known to possess anti-inflammatory and immunomodulating activities. However, the effects and mechanism of action of Ganoderma lucidum on lipopolysaccharide (LPS) and interferon-gamma (IFN-γ)-induced nitric oxide (NO) production and its-related cytokine expression are not yet fully understood. This study aimed to evaluate the effect of Ganoderma lucidum on NO production and NO-mediated pro-inflammatory cytokine expression in LPS/IFN-γ-induced RAW 264.7 macrophage-like cells. Methods and Results : The results showed that Ganoderma lucidum inhibited inducible NO synthase (iNOS) expression of RAW 264.7 macrophage-like cells at non-cytotoxic concentrations probably through the reduction of reactive oxygen species (ROS) production. After pre-treatment of cells with non-toxic doses of Ganoderma lucidum; NO production was significantly decreased. Moreover, Ganoderma lucidum treatment suppressed LPS/IFN-γ -stimulated pro-inflammatory cytokine secretion, including interleukin-1β and interleukin-6, in a dose-dependent manner. Conclusion : Taken together, these results indicate that the anti-inflammatory activation of Ganoderma lucidum in LPS/IFN-γ-stimulated macrophages might be due to abrogation of NO-dependent cytokine release by impairment of iNOS expression via ROS generation.

Background : The roots of Codonopsis lanceolata have been used as a tonic crude drug and an edible plant in Korea. The plant mainly contains triterpenoid saponins, including codonolaside, codonolasideⅠ-Ⅴ, lancemaside A-G. Their saponins have shown anti-inflammatory effects such as bronchitis and cough, insomnia and hypomnesia. C. lanceolata is well known to affect various pharmacological effects for human health, and its consumption is increasing. Recently, plant and plant-derived products were treated a part of the healthcare system by applying the bioactive phytochemicals. Antioxidant and immune activity substances in food play an important role as a health-protecting factor. This study was designed to investigate the in vitro immune cell growth and xanthine oxidase Inhibitory activity of different storage period and storage temperature of C. lanceolata. Methods and Results : The plant materials were used the roots of C. lanceolata cultivated in Jeju area, Korea. Immune enhancing effect was conducted using T cell and B cell of human immune cells. Each cell incubated for 8 days with the sample extracts compared to the control group, and the immune activation was measured according to the growth of immune cells. The xanthine oxidase Inhibitory activity was measured by modifying the method of Noro(1983). In different storage period and storage temperature conditions, the immune cell growth of C. lanceolata extract promoted a concentration-dependent manner in both human T cell and B cell, and did not show a significant difference. The xanthine oxidase Inhibitory activity of C. lanceolata extract tended to decrease more, depending on the longer the storage period or the higher the storage temperature. Conclusion : These results of this study suggested that the root of C. lanceolata may assist in the potential biological activities, and can be used as a source of human health products.

Background : Lutein, a xanthophyll, consists of chains with 8 conjugated double bounds containing closed rings on each end of the chain. This carotenoid is found in fruits and vegetables, especially dark green leafy vegetables such as green tea. In this study, we investigated the anticancer effects of purified lutein from green tea on human cancer cell lines containing prostate carcinoma cancer cells (LNCaP). Methods and Results : Prostate carcinoma cancer cells (LNCaP) were cultured and evaluated the inhibitory effect of lutein isolated from green tea compared other carotenoids (β-carotene and lycopene) on cell proliferation. Cyclin D1 and PCNA were evaluated as cell differentiation. In results, PCNA/cyclin regulates the initiation of cell proliferation by mediating DNA polymerase. Under cultural conditions, lycopene remarkably suppressed the PCNA expression prostate cancer cell line LNCaP in higher doses (20 μM - 100 μM) statistically. However, β-carotene and lutein presented the less inhibitory effects on PCNA expression. Determination of PCNA expression in control and treated cells demonstrates that lycopene did affect proliferation in LNCaP cancer cells in dose-dependent manner. However, β-carotene and lutein suppressed the cyclin D1 expression in dose-dependent manner but no in lycopene group. These results indicate that differ carotenoids presented the various suppressive ability of PCNA and cyclin D1 expression in cell proliferation. Conclusion : In conclusion, lutein suppressed the carcinogenesis of induced prostate cancer cell line by acting as a suppressor for inhibiting the expression of cyclin D.

Background : The objective of this study was to investigate antioxidant activities, inhibitory activities against heme induced colonic epithelial cell proliferations, anti-inflammatory activities and anthocyanin profiles in the anthocyanin rich fraction (ARFAM) from fruits of Aronia melanocarpa, where these are considered functional substances and available food coloring agents in Korea. Methods and Results : Anthocyanins were identified by reversed-phase C18 column chromatography and HPLC-DAD-ESI/MS analysis. To compare the antioxidative and anti-inflammatory capacity of Aronia melanocarpa berries, recognized for their high content of anthocyanins, isolation method was developed to obtain high-purity anthocyanins in the extract. Anthocyanin-rich fractions (ARFAM) enriched in anthocyanins were found to be potent strong inhibitory activity towards heme induced colonic epithelial cell proliferations are associated with an increased risk of colon cancer than acidic ethanol extract (AME). The immunomodulation properties were assessed in growth of both human B and T cells, its cytokines secretion such as IL-6 (interleukin-6) and TNF-α (tumor necrosis factor-alpha). AME enhanced interleukin-6 and reduced tumor necrosis factor-a production, whereas ARFAM only had a effect in increasing of IL-6 expression. Conclusion : These results demonstrated that there was no major relationship between the antioxidative and immunomodulation capacities of AME and ARFAM.

The walls of guard cells have many specialized features. Guard cells are present in the leaves of bryophytes, ferns, and almost all vascular plants. However, they exhibit considerable morphological diversities. There are two types of guard cells: the first type is found in a few monocots, such as palms and corn, and the other is found in most dicots, many monocots, mosses, ferns, and gymnosperms. In corns, guard cells have a characteristic dumbbell shape with bulbous ends. Most dicot and monocot species have kidney-shaped guard cells that have an elliptical contour with a pore at its center. Although subsidiary cells are common in species with kidney-shaped stomata, they are almost always absent in most of the other plants. In this study, there were many different stomatal features that were associated with kidney-shaped guard cells, but not dumbbell shaped guard cells, which are present in most grasses, such as cereals. Each plant investigated exhibited different characteristic features and most of these plants had kidney-shaped guard cells. However, the guard cells of Chamaesyce supina Mold, were often more rectangular than kidney-shaped. In contrast, Sedum sarmentosum guard cells were of the sink ensiform type and in Trifolium repens, the guard cells exhibited a more rhombic shape. Therefore, kidney-shaped guard cells could be divided into a number of subtypes that need to be investigated further.

We investigated the antioxidative and protective effects of corn silk (Zea mays L.) ethanol extracts on human HaCaT cells and erythrocytes. The NICS-2 fraction, extracted from corn silk, exhibited favorable 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activities with IC50valuesof13.3± 0.3 μg/mL and 14.2 ± 0.1 μg/mL when compared with those of α-tocopherol, a positive control, with IC50=10.4± 02.2 and 22.2 ± 3.6 μg/mL, respectively. In addition, we investigated skin protection effects of NICS extracts of corn silk in HaCaT keratinocytes. To investigate the pharmacological potential of NICS-1 and NICS-2 extracts of corn silk on UV-B-induced damage in HaCaT cells, we measured the activity of interleukin (IL) 1a. Our results showed that all the corn silk extracts inhibited the UV-B-induced activity of IL-1a. In particular, NICS-1 extracts of corn silk significantly suppressed IL-1a activity in a dose-dependent manner without inducing cytotoxicity. These results indicate that the ethanol extracts of corn silk (Zea mays L.) could function as natural cytoprotective agents and antioxidants in biological systems, particularly the skin exposed to UV radiation, by protecting cellular membrane against reactive oxygen species (ROS).

This study was conducted to compare the antioxidant, anticytotoxic, and anti-inflammatory properties of Euphorbia maculata ethanol extract with those of E. supina ethanol extract. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical and superoxide scavenging activities of E. maculata at 50 μg/mL were 38.3 ± 3.7 and 21.5 ± 1.2%, respectively, whereas those of E. supina at the same concentration were 109.4 ± 0.9 and 59.5 ± 4.8%, respectively. Oxygen radical absorbance capacities of E. maculata and E. supina at 10 μg/mL were 14.70 ± 0.63 and 26.17 ± 1.36 nmol/mL Trolox, respectively. Cupric reducing antioxidant capacities of E. maculata and E. supina at 10 μg/mL were 10.22 ± 0.97 and 62.99 ± 5.28 nmol/mL Trolox, respectively. Total phenolic contents of E. maculata and E. supina at 50 μg/mL were 29.03 ± 0.14 and 87.89 ± 0.20 nmol/mL gallic acid, respectively. E. maculata and E. supina were reported to prevent supercoiled DNA breakage induced by peroxyl and hydroxyl radicals in a concentration-dependent manner, where protection against the supercoiled DNA breakage provided by E. supina was greater than that provided by E. maculata. E. maculata and E. supina at 100 μg/mL inhibited tert-butyl hydroperoxide-induced cytotoxicity in HepG2 cells by 49.4 ± 4.3 and 87.3 ± 4.5%, respectively. E. maculata and E. supina at 500 μg/mL inhibited lipopolysaccharide-induced nitric oxide production in RAW 264.7 cells by 63.1 ± 7.0 and 85.2 ± 1.6%, respectively. The antioxidant capacities including DPPH radical scavenging, superoxide scavenging, oxygen radical absorbance, and cupric reducing antioxidant activity were found to be highly correlated with total phenolic content (0.896 < r < 0.983, p < 0.01) and anticytotoxic activities (0.915 < r < 0.960, p < 0.01). However, the superoxide scavenging activity was not significantly correlated (r = 0.604, p > 0.05) with the anti-inflammatory activity. Thus, these findings demonstrated that the radical scavenging, anticytotoxic, and anti-inflammatory capacities of E. supina were more potent than those of E. maculata. Further studies are needed to elucidate the properties of polyphenolic constituents in E. supina responsible for these effects and the underlying mechanisms.

본 연구에서는 드럼스틱 잎 추출물과 분획들의 항산화 활성 및 HaCaT 세포와 적혈구 세포에서의 세포보 호효과를 측정하였다. 모든 실험은 드럼스틱 잎의 50% 에탄올 추출물, 에틸아세테이트 분획 및 아글리콘 분획 을 이용하였다. 1,1-Diphenyl-2–picrylhydrazyl radical을 이용한 자유 라디칼 소거 활성(FSC50)은 50% 에 탄올 추출물(77.10 μ g/mL) < 에틸아세테이트 분획(20.63 μ g/mL) < 아글리콘 분획(17.00 μ g/mL) 순으로 증가하였다. 루미놀을 이용한 Fe3+-EDTA/H2O2계에서의 활성산소 소거 활성(총항산화능, OSC50)은 아글리콘 분획의 OSC50 값이 0.63 μ g/mL로 추출물 중 가장 큰 항산화능이 나타났으며, 이는 L-ascorbic acid (1.50 μ g/mL)의 항산화 활성보다 컸다. 1O2로 유도된 적혈구 세포 손상에 있어서 50% 에탄올 추출물 및 아글리콘 분획의 세포 보호 효과(τ50)는 10 μ g/mL에서 각각 46.9 및 122.1 min을 나타냈다. 이는 지용성 항산화제로 알려진 (+)-α-tocopherol (37.7 min)보다도 훨씬 큰 세포 보호 활성을 보여주었다. 400 mJ/cm2의 UVB를 HaCaT 세포에 조사하여 세포손상을 유도한 후 에틸아세테이트 분획 및 아글리콘 분획은 0.20 ∼ 1.56 μ g/mL 농도에서 농도 의존적으로 세포보호효과를 나타내었다. 이상의 결과들은 자외선에 노출된 피부에서 드럼스틱 잎 추출물과 분획들이 ROS 소거를 통하여 세포를 보호함으로서 화장품에서 천연 항산화제로서 사용 가능함을 시사하였다.

Algal blooming in 4 major rivers introduces substantial impacts to water front activity. Concentrations of algae are increasing at major points along the Geum River. Ecosystem food webs can be affected by algal blooming because blue-green algae release toxic materials. Even though there have been many studies on blue-green algae, its causality to environmental factors has not been completely determined yet. This study analyzed the exclusive correlation between various hydrometeorological, water quality, and hydrologic variables and the cell number of cyanobacteria to understand causality of blue-green algae in the Geum River. A prewhitening process was introduced to remove the autocorrelation structure and periodicity, which is useful to evaluate the effective relationship between two time series.

DJ#3과 DJ#13은 항염증질환 소재로서 가능성을 확인하 고자 국내 전통식품 풀질인증 된장에서 장기간 숙성된장으 로 선발된 시료이다. DJ#3과 DJ#13 추출물의 세포독성을 살펴보기 위하여 혈관내피세포를 이용하여 세포의 생존율 을 살펴본 결과 DJ#3과 DJ#13 추출물 모두 100 μg/mL의 농도까지 전혀 독성을 나타내지 않았다. 또한 DJ#3과 DJ#13 추출물의 항염증 효과를 TNF-α에 의해 활성화된 혈관내피세포에서의 NO 생성, 염증관련 단백질 발현과 mRNA 유전자 발현의 변화를 통하여 확인하였다. 혈관내 피세포에 TNF-α를 처리한 결과 NO의 함량이 유의적으로 감소하였다가 DJ#3과 DJ#13 추출물(20, 50, 100 μg/mL)을 처리하였을 때 유의성은 없으나 증가하였다. 세포배양액내 VCAM-1, ICAM-1 발현을 확인한 결과 혈관내피세포에 TNF-α를 처리한 군에서 증가된 VCAM-1 발현이 DJ#3 추출 물 20, 50 μg/mL에서 유의성 있는 감소를 보였다. 또한 DJ#3 추출물은 NO 생성과 연관 있는 eNOS mRNA의 발현을 농도 의존적으로 증가하였으며, iNOS mRNA의 발현은 농 도 의존적으로 감소하였으며 이는 NO 생성 증가가 iNOS의 발현억제를 경유한 것으로 사료된다. 또한 다수의 항염증 약물들의 작용기전이 되는 COX-2의 생성억제를 살펴본 결과 DJ#3 추출물은 TNF-α에 의해 발현되는 COX-2 단백질 의 발현을 억제하였음을 확인할 수 있었다. 또한, DJ#3 추출 물은 CAMs 단백질 및 mRNA 발현율의 감소됨을 보였다, 이상의 결과로 보아, DJ#3 추출물은 혈관내피세포에서 TNF-α로 유도된 혈관염증을 감소하는 효과를 가지고 있으 며, 항염증물질의 연구에 기초 자료로 활용이 가능할 것으 로 기대된다. 또한 염증과 관련된 cytokine 및 단백질 발현 메커니즘에 대한 추가적인 연구가 필요할 것으로 판단된다.

방사선 치료시 최근 많이 쓰이고 있는 IMRT, Tomotherapy, mArc(modulation arc therapy)의 치료법을 선량 적인 측면에서 상호 비교하고자 한다. 비소세포성 폐암의 환자를 대상으로 하여 치료계획용적에 58.0 Gy를 처방선량을 기준으로 설정하였으며 주변 정상장기는 척수, 식도, 간을 설정하였다. PTVmean는 mArc의 경 우 57.60 Gy, Tomotherapy가 61.04 Gy, IMRT는 58.95 Gy이었다. 식도(Esophagus)의 평균선량은 mArc가 2.84 Gy였고, Tomotherapy가 5.14 Gy, IMRT가 1.84 Gy로 나타났다. 간(Liver)은 mArc는 19.44 Gy, Tomotherpy가 12.22 Gy, IMRT는 21.97 Gy이었고 척수(Spinal Cord)은 mArc 5.72 Gy, Tomotherapy가 7.08 Gy, IMRT는 6.15 Gy로 측정되었다. 또한, 선량포함도와 선량체적곡선 등의 자료를 관찰해 본 결과 mArc와 Tomotherapy 그리 고 IMRT의 결과와 현저한 차이를 보이지는 않았다. 하지만, 본 연구는 폐암이라는 질환으로 한정하였었고 실험군의 수가 적은 단점을 가지고 있으므로 좀 더 많은 질환과 환자를 대상으로 연구를 폭넓게 진행 한다 면 환자 맞춤형 치료기법을 개발하여 적용될 것으로 사료된다.

Background: Sedum takesimense Nakai has been used as folk medicine in Korea. The present study aimed to determine the biological activity of S. takesimense by investigating the anti-inflammatory effects of S. takesimense water extract (SKLC) on the lipopolysaccharide-induced inflammatory response in RAW 264.7 cells. Methods and Results: Cytotoxicity of SKLC on RAW 264.7 cells was determinded by performing MTS assay was found to have no cytotoxic effect on RAW 264.7 cells at a concentration range of 62 - 500 ㎍/㎖. Further, pretreatment of SKLC inhibited lipopolysaccharide-induced nitric oxide (NO) production in a dose-dependent manner. To determined the inhibitory mechanisms of SKLC on inflammatory mediators, we assessed the inducible nitric oxide synthase (iNOS) and cyclooxygnease-2 (COX-2) pathways. The activities of these pathways were decreased in a dose-dependent manner by SKLC. The production of tumor necrosis factor- α (TNF-α), interleukin (IL)-1β‚ and IL-6 were also reduced. Conclusions: These results suggest that the down regulation of iNOS, COX-2, TNF-α, IL-1β‚ and IL-6 expression by SKLC are mediated by the down regulation of nuclear factor-κB (NF-κB) activity, a transcription factor necessary for pro-inflammatory mediators. This might be the mechanism underlying the anti-inflammatory effects of SKLC.

피부 기저막(basement membrane, BM)이란 표피와 진피 사이에 존재하는 특별한 구조물로 표피와 진피 를 단단히 고정시켜 피부 구조를 유지하는 데에 중요한 역할을 수행한다. 노화 및 자외선 노출에 의한 피부 기저막 의 구조적 변화와 파괴는 피부 주름 형성과 탄력 저하를 포함하는 피부노화 현상의 요인으로 여겨지고 있다. Laminin-332 (LN-332)는 피부 기저막을 구성하는 주성분으로 피부에서 표피와 진피를 단단히 고정시키는데 중요한 역할을 한다. 본 연구에서는 왕불유행 헥산 분획물(Melandrium firmum hexane fraction, MFHF)이 각 질형성세포에서 LN-332 발현에 미치는 효과를 확인하였다. 정량적 real-time PCR (RT-PCR)과 단백질 발현 분석을 통해서 MFHF가 LN-332의 mRNA 발현 및 단백질 발현을 촉진시키는 것을 확인하였다. 또한 MFHF가 어떤 신호전달 경로를 통해 LN-332 발현을 조절하는지 확인하기 위하여 p38 MAPK 억제제인 SB202190과 ERK1/2 억제제인 U0126을 처리한 결과, p38 MAPK 억제제에 의해서 LN-332 발현이 완벽히 억제됨을 확인 하였다. 또한, 피부 기저막을 구성하고 있는 콜라겐 타입 VII과 integrin α6의 mRNA 발현 역시 MFHF에 의해 증가하는 것을 확인하였다. 우리는 본 연구를 통해 MFHF가 각질형성세포에 작용하여 피부 기저막을 구성하는 성분들의 생성을 촉진할 수 있는 소재로 작용할 수 있다는 것을 확인하였다. 이러한 결과는 기저막의 구조적, 기능 적 이상에 의해 나타나는 피부노화 현상의 개선을 위해 활용할 수 있을 것이라 제안한다.

피부의 가장자리에 위치한 표피는 개체의 생존에 절대적으로 중요하며, 기저막에 위치한 표피줄기세포에 의해 평생 끊임없이 재생되고 있다. 표피줄기세포는 한정된 세포분열을 진행하고 각질세포로 분화하는 transit amplifying (TA)세포를 만들어 냄으로써 오랜 기간 재생에 필요한 많은 각질세포를 만들어 낼 수 있다. 본 연구 에서는 표피줄기세포 세포분열 활성화 화합물로 목단으로부터 추출한 페오놀(paeonol)을 350여 개의 화합물들 로부터 검색해 냈다. 이러한 세포분열 활성화 효능은 표피줄기세포 특이적으로 나타나며, 표피줄기세포의 지표 로 알려진 p63 단백질의 발현 경향은 페오놀을 처리한 표피줄기세포에서 변화하지 않음을 유세포분석법으로 확인하였다. 콜로니형성 분석에서는 페오놀을 처리한 표피줄기세포가 1.3배 이상 더 나은 콜로니 형성능을 보여 주었다. 그리고 PCR array 분석을 통해서 페오놀의 효능은 여러 신호전달에 의해 나타남을 알 수 있었다. 이러 한 결과들로부터 페오놀이 줄기세포성에 영향을 주지 않고 표피줄기세포의 재생능력을 향상시키므로 표피줄기 세포 활성화 물질로서 화장품 소재로 적용될 수 있을 것으로 판단된다.

Background : The purpose of the present investigation is to enhance extracellular acidic protease production by subjecting a protease producing strain Cordyceps pruinosa DK-01 to random mutagenesis by UV irradiation after ethidium bromide treatment. Methods and Results : Mutants were screened as protease producers on the basis of zone of clearance and relative proteolytic activity (RPA) on skimmed milk agar plates. In addition, mutants showed strong pink-red color intensity and different RAPD profiling compared with wild type control. Four mutants were randomly selected and their extracellular enzyme activities were investigated. In liquid culture without casein, 2.2-, 2.9-, 5.2- and 4.4-fold higher acid protease activity was achieved from mutants DK-m9, -m11 and -m12, respectively, than that of wild type strain (11.13 ± 1.60 U/ml). In liquid culture with casein, 1.1-, 1.3-, 1.3 and 1.3-fold higher acid protease activity was achieved with those mutants were found to produce, respectively, than that of wild type strain (93.95 ± 12.84 U/ml). Maximum acid protease activity was noticed from a mutant DK-m11 in liquid culture with casein (121.18 U/ml) and without casein (57.65 U/ml). The extracellular acid protease produced from DJ-m11 that was active in the pH range 4.5-6.5 and optimum temperature for the activity was 37°C. Furthermore, we found a deformed, shorten structure of setae on the elytron surface of dynastid beetles treated with culture supernatant of the DK-m11. Conclusion : These findings have more impact on enzyme economy for biotechnological and insecticidal applications of fungal proteases.

Background : Lectins were individually isolated from natural Korean mistletoe (nML) and its in vitro cultured calli (cML). Both of the lectins showed the difference in bioactivities such as cytotoxicity and cytokine induction. Methods and Results : Target cells (1 x 104 cells/well) were seeded independently into each well of a 96-well culture plate and incubated with different concentrations of each lectin. Survivability of target cells was determined by CCK-8 kit (Sigma, USA) according to manufacturer’s directions. The nML showed 46, 34 and 5.5 times stronger than cML in cytotoxicity (IC50) to human melanoma cell line (SK-MEL-28), human carcinoma cell line (NCI-H1650) and murine macrophage (RAW 264.7), respectively. In addition, respective lectins directly stimulated macrophage RAW 264.7 but they showed the difference in enhanced productivity of some inflammatory cytokines. Compared with cML, the nML induced both TNF-α and IL-1β at its low concentration. Administration of two kinds of lectins (10-1000 μ g/kg body weight) to ICR mouse did not show any significant changes on the level of alanine transaminase (ALT), glutamate-oxaloacetic transaminase (GOT) and blood urea nitrogen (BUN) in sera. Conclusion : From the above results, we may suggest that nML and cML showed differences in cytotoxic effects and cytokine production due to the difference in amounts from sources.