An assessment was made of beta-site amyloid precursor protein (APP) cleaving enzyme (BACE1) inhibitory, feeding, climbing activities and lifespan of the diarylalkyls curcumin (CCN), demethoxycurcumin (DCCN) and bisdemethoxycurcumin (BDCCN) identified in the rhizomes of Curcuma longa. Based on IC50 values, BDCCN (0.024 mM) was the most inhibitory constituent, followed by DCCN (0.31 mM) and CCN (0.59 mM). Overall the three curcuminoids were significantly less inhibitory than BACE1 inhibitor IV isophthalamide (8.5 × 10-5 mM). The expression of human APP and BACE1 in compound eye of Drosophila melangaster presented rough abnormal ommatidial lattice. Co-expression of APP and BACE1 within the developing nervous system of drosophila showed climbing defects. These transgenic flies kept on media containing 1 mM of CCN and BDCCN were observed to ameliorate eye degeneration, significantly suppress locomotive dysfunctions, and increase media life time, as well as isophthalamide. CCN and BDCCN as human BACE1 inhibitory constituents may be used as potential therapeutics or lead molecules to develop Alzheimer's disease treatment drugs.

비록 천연유래물질 중의 하나인 통초가 항산화작용, 항염증작용 및 해열작용과 같은 효능이 있다고 알려져있지 만, 종양에서의 암예방효능에 대한 연구는 보고된 바가 없 다. 본 연구에서는 사람 자궁경부암세포주인 HEP-2세포에서 통초메탄올추출물의 증식억제 효능 및 관련 분자표적을 확인하고자 하였다. 통초메탄올추출물은 세포증식을 유의성있게 억제하고, 세포사멸을 유도하는 것으로 나타났다. 또한, Bid의 발현에 영향을 주어 truncation을 유도하고, 이로 인해 cytochrome c가 마이토콘드리아에서 세포질로 이 동하도록 유도하였지만, Bid 이외의 다른 Bcl-2 family 에는 영향을 주지 못했다. 따라서, 이러한 결과를 종합해 볼 때, 통초메탄올추출물은 자궁경부암에서 암예방효능을 가진 잠재성있는 천연추출물이 될 수 있을 것으로 사료된다.

Cordycepin, a specific polyadenylation inhibitor, is the main functional component in Cordyceps militaris that is one of the top three renowned traditional Chinese medicines. In this study, we performed in vitro experiments to investigate the anti-invasive and anti-metastatic activities of cordycepin using human prostate carcinoma LNCaP cells. Cordycepin were administered and their effects on LPS-induced cell migration and invasion by wound healing migration assay, measurement of TER and In vitro invasiveness assay. Within the concentrations which were not cytotoxic effects, cordycepin caused a concentration-dependent suppression of LPS-induced cell migration and invasion. The anti-invasive activity of cordycepin was also found to be associated with increased tightness of the TJ, which was confirmed by an increase in TER. The activity of MMP-2 in LNCaP cells was dose-dependently inhibited by treatment with cordycepin, and this was also correlated with a decrease in expression of its mRNA and proteins, and up-regulation of TIMPs expression. Additionally, cordycepin repressed the LPS-induced NF-kB activation and phosphorylation of PI3K/AKT. Taken together, these findings suggest that cordycepin inhibited LPS-induced migration and invasion of LNCaP cells by down-regulating the expression and activity of MMP-2, and the possible targets may be NF-kB and PI3K/AKT.

The aim of this study was to investigate the antitumor activity of solvent fractions from Auricularia auricula-judae 70% ethanol extract and confirmed the active components of dichloromethane fraction showing a potent antitumor activity than other fractions in the broncheoalveolar and gastric cancer cells. The solvent fractions of Auricularia auricula-judae extract, inhibited the growth proliferation of tumor cells in dose-dependent manner. The principle components of dichloromethane fraction were 5,11,17,23-tetrakis (1,1-dimethyl)-28-methoxypentacyclo [19.3.1.1 (3,7).1 (… (65.85%) and diazane (6.17%). The antitumor active components, diazane and gibberellic acid (GA3) were identified in this fraction by GC-MS analysis and lower antitumor activities than dichloromethane fraction. The unknown components of dichloromethane fraction were responsible for its cytotoxic effects on tumor cells. Based on IC50 value, gibberellic acid was little cytotoxic itself. According to PCR amplification, the apoptosis of tumor cells were induced by the down-regulation of Bcl-2 and over-expression of P53 on the presence of solvent fractions, diazane and gibberellic acid. Thus, these findings suggest that the dichloromethane might be used as functional feed additive that suppress the tumor growth in the body than other solvent fractions of Auricularia auricula-judae extracts.

To evaluate the effect of spermatozoa culture on glycosidase activity of frozen-thawed spermatozoa in human, the spermatozoa were treated experimentally and assayed for activities of α-L-fucosidase, α-D-mannosidase, β-D-galactosidase and N-acetyl-β-D-glucosaminidase (β-GlcNAc’ase). The β-GlcNAc’ase activity was at least two-folds higher than other glycosidases regardless of spermatozoa incubation (p<0.05). The spermatozoa motility was decreased with incubation periods, but no effects by different glycosidases on the changes of spermatozoa motility during the various periods of incubation. In all glycosidases, the spermatozoa-zona binding rates in spermatozoa without incubation were higher than in spermatozoa incubated for 2 h (p<0.05). β-GlcNAc’ase is present mainly in the plasma membrane of spermatozoa frozen-thawed in human. It was also shown that the glycosidase activity was increased in all glycosidases in spite of lower sperm-zona binding by spermatozoa incubation.

Erythropoietin (EPO) is a glycoprotein hormone secreted from primarily cells of the peritubular capillary endothelium of the kidney, and is responsible for the regulation of red blood cell production. We constructed and expressed dimeric cDNAs in Chinease hamster ovary (CHO) cells encoding a fusion protein consisting of 2 complete human EPO domains linked by a 2-amino acid linker (Ile-Asp). We described the activity of dimeric hyperglycosylated EPO (dHGEPO) mutants containing additional oligosaccharide chains and characterized the function of glycosylation. No dimeric proteins with mutation at the 105th amino acid were found in the cell medium. Growth and differentiation of the human EPO-dependent leukemiae cell line (F36E) were used to measure cytokine dependency and in vitro bioactivity of dHGEPO proteins. MTT assay at 24 h increased due to the survival of F36E cells. The dHGEPO protein migrated as a broad band with an average molecular mass of 75 kDa. The mutant, dHGEPO, was slightly higher than the wild-type (WT) dimeri-EPO band. Enzymatic N-deglycosylation resulted in the formation of a narrow band with a molecular mass twice of that of of monomeric EPO digested with an N-glycosylation enzyme. Hematocrit values were remarkably increased in all treatment groups. Pharmacokinetic analysis was also affected when 2.5 IU of dHGEPO were intravenously injected into the tails of the mice. The biological activity and half-life of dHGEPO mutants were enhanced as compared to the corresponding items associated the WT dimeric EPO. These results suggest that recombinant dHGEPO may be attractive biological and therapeutic targets.

The present paper deals with an analysis of relationship among technical human resources management as a precursor, technology innovation activity and achievements of technical innovation on the basis of preceding empirical studies on technology innovation activities of manufacturing corporations. The analysis shows that First, the technical human resources management is found to have influence upon technology innovation activity in various ways, implying that the role of technical human resources management as a key to technical innovation is most important of all to enable manufacturing companies to gain edge in competition by means of technology innovation activity; and Second, technology innovation activity exercises impacts on the achievements of exploitative technology innovation as well as on the achievements of exploratory technology innovation on the part of manufacturing industry. The above findings prove that the level of technology innovation activity may be a source for superior competitiveness of manufacturing business as a result of technology innovation performance. Manufacturing corporations, thus, need to place more weight on stepping up their executive level of technology innovation activity factors than on increasing simply the level of technical investment.

Periodontal disease is a major oral disorder and comprises a group of infections that lead to inflammation of the gingiva and the destruction of periodontal tissues. PPARy plays an important role in the regulation of several metabolic pathways and has recently been implicated in inflammatory response pathways. However, its effects on periodontal inflammation have yet to be clarified. In our current study, we evaluated the anti-inflammatory effects of PPARy on periodontal disease. Human gingival fibroblasts (HGFs) treated with lipopolysaccharide (LPS) showed high levels of intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), matrix metalloproteinase-2 (MMP-2), and -9 (MMP-9). Moreover, these cells also showed upregulated activities for extracellular signal regulated kinase (ERK1/2), inducible nitric oxide synthase (iNOS) and cyclooxygnase-2. However, cells treated with Ad/PPARy and rosiglitazone in same culture system showed reduced ICAM-1, VCAM-1, MMP-2, -9 and COX-2. Finally, the anti-inflammatory effects of PPARy appear to be mediated via the suppression of the ERK1/2 pathway and consequent inhibition of NF-kB translocation. Our present findings thus suggest that PPARy indeed has a pivotal role in gingival inflammation and may be a putative molecular target for future therapeutic strategies to control chronic periodontal disease.

This study examined the effects of red light generated from a light emitting diode (LED) upon proliferation and mitochondrial stress in human gingival fibroblasts (hGFs). Cells were exposed to LED-generated red light at a clinically relevant intensity and distance with a 610-630 nm wavelength for various times (0-48 min). At different exposure times, cells were processed for the analysis of succinate dehydrogenase (SDH) activity, proliferation, mitochondrial membrane potential (MMP) and cytotoxicity. Cell cycle progression was also investigated by flow cytometry after staining with propidium iodide. Red light exposure was found to inhibit SDH activity and DNA synthesis in hGFs in a time-dependent manner. Light exposure also reduced the MMP levels in these cells and this was closely associated with a G0/G1 arrest. In contrast, exposure of hGFs to red light for 48 min led to a dramatic loss of MMP with an attendant increase in cytotoxicity. These findings demonstrate that LED-generated red light may cause mitochondrial stress and growth inhibition in hGFs during tooth whitening therapy, depending on the length of the exposure.

목적 : 연구의 목적은 우리나라 노인들의 문화와 정서에 적합한 활동척도를 개발하여 남녀 노인의 활동 수준과 의미 있는 활동을 비교하고, 노인의 삶의 질에 영향을 미치는 요인들의 관련성을 규명하였다. 연구방법 : 연구의 대상은 60세 이상 남자 노인 199명, 여자 노인 258명으로 총 457명이 참여하였으며, 노인생활만족척도를 사용하여 노인의 삶의 질을 평가하였다. 또한 노인의 활동수준은 평가를 위해 한국판 활동척도를 개발하여 연구에 이용하였다. 결과 : 노인의 수행 활동을 성별로 비교하여 볼 때 여자 노인은 수단적 일상생활을 남자 노인은 고강도 신체활동을 더 많이 하였으며, 노인의 활동과 삶의 질과의 상관관계를 통하여 남, 여, 전체 노인에서 수단적 일상생활, 저강도 신체활동, 고강도 신체활동, 사회활동에 많이 참여할수록 삶의 질은 높았다. 최종적으로 위계적 다중선형회귀분석을 통하여 노인의 삶의 질에 영향을 주는 요인으로는 전체 노인의 경우 연령, 건강상태, 생활수준, 노후설계 준비기간, 사회활동이, 남자 노인의 경우 건강상태, 배우자 유무, 생활수준, 노후설계 준비기간이 삶의 질을 예측할 수 있는 요인으로 작용하였으며, 여자 노인의 경우 연령, 건강상태, 생활수준, 노후 설계 준비기간, 사회활동이 삶의 질을 예측할 수 있는 요인으로 작용하였다. 결론 : 연구 결과를 토대로 남녀 노인에 적합한 성별에 따른 차별화된 활동을 개발하고 1차, 2차, 3차 예방 사업에 사회활동 요소를 포함하여야 할 것이다. 향후 전국 조사를 통하여 우리나라 노인들을 대상으로 활동 수준과 삶의 질과의 관련성을 규명할 수 있는 다양한 연구들이 수행되어야 할 것이며, 작업치료적 관점에서 장애 노인을 대상으로한 활동의 변화와 삶의 질에 대한 연구가 진행되어야 할 것을 제언한다.

Laser is used to prevent the early dental caries in dental f ield and to apply for treatment of stomatitis and hyper sens it ivity , and laser mass Recently it is reported that laser i1'r adiation affect on soft tissue treatment and bone 1'emodelling after dental implantation. The purpose of this study was to examine laser irradia ti on effect on activity of normal human osteoblast on titanium plate in vitro by various laser wave length, and to observe morphologic change of NHost on LiLa nium plate and to a nalysis concentration of Ca"++ , IP and ALP NHost were cultured in DMEM containing 10% FBS, and observed by in verted microscope 1"or attatchment to the surface of titanium plate. Ca ++, I.P. , and a lkaline phosphat ase(ALP) concent ration in medium was calculated during 4 weeks, which was treated with Wilcoxon rank, Anova test and linear regression. The obtained results were as follows Morphologic changes showed rapid growth rate of NHost ++ at 3 days of laser lrradiatlOn ln spite of laser wave type, Ca" and P concentration was decreased at 2 weeks and was the hig hest at 3 weeks, but decreased at 4 weeks In spite of laser wave type, ALP concentration was decreased at 2 weeks but was increased at 3-4 weeks, From the aboving results, in spite of laser wave type, there were rapid growth rat e of NHost a nd no significant of Ca"++ , IP and P concen tr ation but these concentration showed predominant change than that of control

The present study was conducted to investigate gelatinolytic activities in HAM and to determine whether there are any changes in gelatinolytic activity profiles when the cells are cultured in hepatogenic medium. Placenta was obtained during caesarean section of the volunteers, with informed consent. HAM were isolated from amniotic membrane using collagenase type A HAM were cultured in hepatogenic medium for 3 weeks and the conditioned media were obtained at day 7, 14 and 21. The zymographic pattern of gelatinolytic activity of the HAM did not undergo a change during passages. When the HAM were cultured in a fibronectin-coated dishes in a hepatogenic medium, there was no significant difference of the gelatinase pattern between before and after culture. However, when bFGF was added to the culture, a dramatic increase of 62kDa and 59kDa gelatinases was observed. Interestingly, when ITS instead of FN was present, HAM-conditioned medium also showed a similar increase of both gelatinases. Immunoblotting analysis demonstrated that both 62kDa and 59kDa gelatinases were the active form of MMP-2 resulting from the turnover of MMP-2 proform. Futher study will be necessary to determine the relationship between bFGF and active MMP-2 during hepatogenesis of HAM.

The hematopoietic growth factor erythropoietin (EPO) is required for the maintenance, proliferation, and differentiation of the stem cells that produce erythrocytes. To analyse the biological activity of the recombinant human EPO (rec-hEPO), we have cloned the EPO cDNA and genomic DNA and produced rec-hEPO in the CHO cell lines. The growth and differentiation of EPO-dependent human leukemic cell line (F36E) were used to measure cytokine dependency and in vitro bioactivity of rec-hEPO. MIT assay values were increased by survival of F36E cells at 24h or 72h. The hematocrit and RBC values were increased by subcutaneous injection of 20 IU (in mice) and 100IU(in rats) rec-hEPO. Hematocrit values remarkably increased at 13.2% (in mice) and 12.2% (in rats). The pharmacokinetic behavior with injection of 6 IU of rec-hEPO remained detectable after 24 h in all mice tested. The highest peat appeared at 2h after injection. The long half-life of rec-hEPO is likely to confer clinical advantages by allowing less frequent dosing in patients treated for anemia. These data demonstratethat ree-hEPO produced in this study has a potent activity in vivo and in vitro. The results also suggest that biological activity of ree-hEPO could be remarkably enhanced by genetic engineering that affects the potential activity, including mutants with added oligosaccharide chain and designed to produce EPO-EPO fusion protein.

The flavin-containing monooxygenases (FMOs) (EC 1.14.13.8) are NADPH-dependent flavoenzymes that catalyze oxidation of soft nucleophilic heteroatom centers in a range of structurally diverse compounds including foods, drugs, pesticides, and other xenobiotics. In humans, FMO1 appears to be the predominant form expressed in human fetal liver. cDNA-expressed human FMO and human liver microsomal FMO have been observed to N- and S-oxygenate nucleophilic nitrogen- and sulfur-containing drugs and chemicals, respectively. In the present study, FMOl can be expressed in the baculovirus expression vector system at level of 2.68 nmol FMO1/㎎ of membrane protein. This isoform was examined for its capacity to metabolize methimazole to its S-oxide using thiocholine assay. Kinetic studies of its S-oxide by recombinant human FMO1 result in K_m of 7.66 μM and V_(max) of 17.79 nmol/min/㎎ protein.