Earticle

Lignin, the second most abundant natural compound after cellulose, is a high-molecular weight polymer of phenolic compounds that occurs naturally in plants. On the other hand, lignin is only used by fuel of pulp and paper production process. Many researchers have made efforts on a variety of applications of lignin. In this work, we conducted experiments on the ultraviolet blocking and antioxidant effect of many kinds of commercially available lignin: lignosulfonic acid (sodium salt), lignosulfonic acid (calcium salt), lignin alkali, lignin alkali (low sulfonate content), and organosolv lignin. In order to improve the lignin properties, lignin was degraded by acidolysis and derivatization followed by reductive cleavage (DFRC) method. We have examined ultraviolet absorption ability using an UV-Visible spectrophotometer. The absorbance of lignin samples was measured at 280 nm, 300 nm, and 340 nm. As a result, organosolv lignin showed the highest absorbance of the tested commercial lignin. In addition, we carried out DPPH (2,2-diphenyl-1-picrylhydrazyl) assay and Folin-Ciocalteu (FC) assay in order to examine antioxidant properties of lignin. The results of two different assays have been compared.

Zebrafish was tested as a animal model for anti-obesity. Zebrafish is very useful for animal model because it's very easy for getting its eggs which is very fast for germination and maturation within 48 hrs. Zebrafish was showed its novelity for controlling signal molecules through RT-PCR. This zebrafish model is very convenient for antiangiogenesis and anti-obesity model including zebrafish egg model. Zebrafish, egg, anti-angiogenesis, anti-obesity. in vivo model, primer, β-catenin, VE-cadherin, PI3K, VEGFR2, signal molecule

In this study, the effects of the FK506-mPEG on immune cell activity, skin grafting rejection, and Freund's complete adjuvant arthritis were investigated. The proliferation of T cells was inhibited with increase with the FK506 and FK506-mPEG concentrations. FK506 and FK506-mPEG at concentrations between 0.01 nM and 1000 nM had very similar effects on the proliferation of the T cells. On the other hand, in the case of the proliferation of T cells by calcium ionophore A23187 (1 μM), when the FK506-mPEG concentration was increased from 0.01 to 1000 mM, the proliferation was decreased from 90.8 to 40.3%. This was 1.8-fold higher than that of paramethoxyamphetamine (PMA). The inhibitory effect of FK506-mPEG on mast cell proliferation was higher than that of FK506. When B cells were cultured for 7 days in basal medium with no pokeweed mitogen (PWM), the IgG production was 156.2 ng/ml. On the other hand, in the case of the same treatment with 0.25% of PWM, it was 876.4 ng/ml. This is about 5.6-fold higher than with no PWM. These results show that FK506-mPEG may be practically applicable as a prodrug for the immunosuppressant FK506.

4 kinds of natural product, Anthrisci Radix, Psoraleae Semen, Siegesbeckiae Herba, Corni Fructus were selected as a anti-obesity subatance through anti-angiogenesis effect. Each natural product inhibited signal pathway such as VEGFR2, or PI3K, or β-catenin or VE-cadherin, and all 4 kinds of selected natural products prohibit Akt signal molecule which is down-located thanVEGFR2, PI3K, β-catenin and VE-cadherin. Therefore 4 kinds of natural product can supress signal to NF-kB for not occuring angiogenesis.

10 kinds of natural products bearing anti-angiogenesis and anti-obesity were selected through tube formation and RT-PCR. For verifying the mechanism of anti-angiogenesis, forward and reverse primers of signal molecules such as VEGFR2, PI3K, β-catenin, VE-cadherin were constructed based on NCBI data base. Agarose gel bands showed antiangiogenic effect through variation of signal molecule with increasing concentration of natural products. Also, SREBP-1c, PPARγ and C/EBPα protein which stimulate lipogenesis and adipogenesis were experimented for controlling effect by selected natural products.

Here we report a novel non-antibody protein scaffold developed based on the fold of kringle domain (KD) derived from plasminogen. Based on the sequence analysis of 39 unique KD in 31 human proteins, we designed KD libraries with a rationale to conserve the critical residues for the KD fold, but to introduce mutations which are exposed to the surface in the flexible loops, using synthetic shuffling technique. For target proteins as a proof-ofconcept, we used death receptor 4 (DR4), DR5 and TNFa. Isolated KD mutants against the individual target proteins, using yeast surface display techniques combined with FACS, were solubly expressed well and specifically bind to the targeted proteins with affinity of 10-6-10-8 M, without significant cross-reactivity, assessed by ELISA and SPR. Some KD mutants selected against DR4 or DR5 induced apoptosis in several carcinoma cell lines as a single agent. Also, some KD mutants isolated against TNFa efficiently protected TNFa-induced cell death by neutralizing the cytotoxicity of TNFa. Our results suggest that a novel non-antibody scaffold based on KD fold can be developed as a specific binder to given targets to modulate the biological activity.

To improve the efficiency of photodynamic theragnosis, folate receptortargeted, water-soluble pheophorbide a loaded nanostructured lipid cargo (NLC) was designed and prepared using cacao butter and oleic acid matrix as the core, and pullulan-folate, as the shell material and surfactant. This kind of multifunctional nanoparticles stay stable with no luminescence through blood circulation and accumulate at tumor sites by virtue of their enhanced permeability and retention(EPR)effects, selectively enter cancer cells via folate-mediated endocytosis while sparing normal cells. As the break down of the PF-NLC, membrane fusion of the endosome,and nuclear transport via lipid vehicle will provide a high concentration of pheophorbide a in cancer cell nucleus. After irradiation, a near-infrared fluorescent imaging and nucleus targetted photodynamic destruction will happen simultaneity.

NAD+-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) catalyzes NAD+-dependent oxidation of prostaglandins as well as other nonprostanoid compounds.Growing evidence demonstrates that the COX-derived prostanoids play a critical role in mediating multiple cellular processes such as cell proliferation, differentiation, and apoptosis and in controlling body homeostasis. Especially, the COX-derived prostaglandins have important roles in the pathophysiology of the kidney and ulcer. Inhibitors of this enzyme may be of value in determining the utility of these compounds in hypertension or ulcer. Previously, a series of benzylidene thiazolidinediones with varied ring structure and methylene bridge to phenyl ring through ether linkage were synthesized and assayed for inhibitory activity. It was found that compound CT-8 (5-[4-(cyclohexylethoxy)benzylidene]-2,4-thiazolidinedione) was the most potent inhibitor effective at nM range. Kinetic studies revealed that inhibition by this compound was non-competitive with respect to NAD+ and uncompetitive with respect to PGE2 indicating that the inhibitor interacts with the enzyme at a site distinct from the substrate binding site. This regulatory site appears to overlap with the activator site occupied by imipramine since activation of the enzyme by this activator is competitively inhibited by compound CT-8.

The epidermal growth factor receptor (EGFR) is a member of the ErbB family of transmembrane tyrosine kinase receptors, which includes ErbB1 (or HER-1, or EGFR), ErbB2 (or HER-2/neu), ErbB3 (or HER-3), and ErbB4 (or HER-4) [1, 2]. The expression of EGFR is common in a number of normal epithelial tissues, but in several solid tumors EGFR was overexpressed. For example, in NSCLC (non-small-cell lung cancer), overexpression of EGFR has been reported to be present in over 50% of cases in several series. Several EGFR inhibitors have been developed in recent years, which can be mainly categorized into two classes: small molecules that are inhibitors of the intracellular tyrosine kinase domain by interfering with autophosphorylation by adenosine triphosphate (ATP), and monoclonal antibodies to the extracellular domain of the EGFR [1, 2]. A mouse mAb (A431-96) was raised against EGFR-ECD (extracellular domain). The mAb efficiently inhibited proliferation of A431 cells, which overexpress EGFR on the cell surface and this mAb (A431-96) binds EGFR with affinity of 5.7 x 108 M-1. Here we describe epitope mapping of the mAb in domain level. In competition methods, we used Erbitux and EGF as a competitor which already known the binding sites. Furthermore, using yeast surface display of EGFR fragments [3], we identified the domain-level epitope of mAb which is in the domain III of EGFR-ECD.

Variable glycosylation of antibodies occurs at Asn297 in the Fc region, specifically terminal galactosylation and sialylation. These variations may affect both pharmacokinetic behavior and effecter function, such as complement-dependant cytotoxicity (CDC).In this study, glucose, as a carbon source in culture medium, was substituted for the other sugars in an attempt to induce greater in vivo efficacy of recombinant antibodies and increase their potential as therapeutic drugs by altering glycosylation of antibody. Suspension cultures of recombinant CHO cells were grown in protein-free media, and purification and analysis were performed using liquid chromatographic methods. The results demonstrated that glucose and mannose promote cell growth and lactate production, while galactose and fructose were found to be good energy sources for glycosylation. So, it was proven that the more lactate was produced, the less glycosylation of recombinant antibody was progressed. However, there was a synergistic effect on glycosylation when mannose was used with other sugars.In summary, carbon sources in culture medium can affect glycosylation of recombinant antibodies. These results provide a reference point for the development of therapeutic recombinant antibodies in the industry.

Though the activation of c-Jun NH2-terminal kinase (JNK) has been reported to be essential forautophagic cell death in response to various stressors, the molecular links between JNKactivation and autophagic cell death signaling remain elusive. Here we report that, in the JNKdependent autophagic cell death of HCT116 cells induced by an agonistic antibody against human death receptor 5, JNK activation upregulated Beclin-1 expression and induced Bcl-2 and p53 phosphorylation, suggesting that JNK activation and its consequent fluxes into the proautophagic signaling pathways contribute to the autophagic cell death in cancer cells.

Atherosclerosis is a chronic disease of the arterial wall where both innate and adaptive immunoinflammatory mechanism are involved. Bee venom (BV) and melittin (Mel) have frequently been used as a remedy for inflammatory diseases. We studied whether BV and Mel have antiatherosclerotic effects. Atherosclerotic lesions were induced by lipopolysaccharide (LPS, conc. 2 mg/kg) intraperitoneally injection mice. Mice were fed with an atherogenic diet containing 15% fat, 1.25% cholesterol and 0.5% cholic acid. BV and Mel treatment significantly reduced biochemical damage (total cholesterol, triglyceride levels) and significantly increased high-density lipoprotein (HDL) cholesterol levels in atherosclerosis mice. Furthermore, BV and Mel decreased the expression of tumor necrosis factor (TNF)-α and interleukin (IL)-1β, vascular cell adhesion molecule (VCAM)-1, intercellular adhesion molecule (ICAM)-1, fibronectin and transforming growth factor (TGF)-β1 in atherosclerosis mice. These findings suggest that the anti-atherosclerotic effect of BV and Mel in atherosclerosis mice and BV and Mel may have a protective effect against chronic inflammatory diseases.

Encapsulation of functional proteins inside hydrophilic core space of vesicular liposomes is an important issue in such applications of targeted delivery of therapeutic proteins, immobilized reactions of industrial enzymes [1], and bionano sensing of diagnostic proteins. The encapsulation yield is generally quite low and needs to be improved for the applications to be industrially viable. Several parameters are known to affect the yield [2]; liposomal lipids types, liposome size, buffer type, pH and ionic strength, protein size (or, molecular weight). Enzymes were selected as an encapsulant because it can replace protein drug due to physical and chemical similarity and enable to measure bioactivity. In this study, by fixing the liposomal lipid as DPPC (dipalmitoylphosphatidylcholine) and using various sized proteins as model proteins (i.e., trypsin, 15 kDa; horseradish peroxidase, 44 kDa; hyaluronidase, 80 kDa; enterokinase, 250 kDa), we evaluated the effects of these parameters on encapsulation yield. After encapsulation process, size exclusion chromatography (Sepharose 4B) was applied to separate liposomes and free proteins [3]. Encapsulation yield was calculated in two units; volumetric encapsulation (mole of protein per unit volume) and surface encapsulation (mole of protein per unit surface area). Effects of buffer pH and salt concentration on proteinlipid interaction were measured by surface plasmon resonance. 5 mM of DPPC liposome dissolved 6 ml of 1mg/ml of enzyme solution represented over 50% of encapsulation yield. These encapsulation properties demonstrated optimal condition to achieve high encapsulation yield with more than 80% of initial bioactivity.

Apolipoprotein A-1 (apoA-I) forms discoidal membrane structure with phospholipids during the formation of high density lipoprotein (HDL) and plays a role in reverse cholesterol transport pathway. The discoidal membrane nanostructure, so called Nanodisc, can be easily assembled in vitro using apoA-I and phospholipids. As well as this easy assembly process, the small size, Sushi-like structure, particle size uniformity, and robustness in solution of the Nanodisc gives us a sense of various ways of biotechnological applications, which might be drug delivery system, membrane protein host or therapeutics for cardiovascular diseases. Coenzyme Q10 (CoQ10) is a fat-soluble, vitamin-like substance found in the cells of many organisms. It is used as a nutritional supplement and is beneficial in the treatment of human conditions. [1; 2]. The use of CoQ10 is also gaining popularity in the cosmetic industry owing to its antioxidant properties.In spite of the versatility of CoQ10, the molecule's long side chain of 10 isoprenoid units make CoQ10 poorly water-soluble, limiting its bioavailability and more various applications. In the present study, a novel CoQ10 nanoparticle was assembled by encircling isoprenyl chains with human apolipoprotein A-I protein (apoA-I). This nanostructure of CoQ10 is quite reminiscent of nascent high density lipoprotein (HDL) particle composed of phospholipids and cholesterol.

There is currently increased interest in the identification of antioxidant compounds that are pharmacologically potent and have low or no side effects. Plants produce significant amounts of antioxidants to prevent the oxidative stress caused by photons and oxygen, therefore they represent a potential source of new compounds with antioxidant activity. FARFARE FLOS (FF) has been frequently used on the respiratory system including bronchitis, phthisis. In previous study, the antioxidant activity of extract from FF was studied in vitro methods by measuring the antioxidant activity by TEAC, measuring the scavenging effects on ROS and RNS as well as measuring the inhibitory effect on Cu2+-induced human LDL oxidation. The FF extracts were found to have a potent scavenging activity, as well as an inhibitory effect on LDL oxidation. And this study was designed to evaluate whether FFEA (ethyl acetate fraction of FARFARE FLOS) may ameliorate oxidative stress and inflammatory status through the antioxidative mechanism in LPS-stimulated RAW 264.7 murine macrophage cell line. Treatment of RAW 264.7 cells with FFEA significantly reduced LPS-stimulated ROS generation, NO production, iNOS mRNA level, expression of proinflammatory cytokine such as IL-1β, IL-6, TNF-α and HO-1 in a dose-dependent manner. In conclusion, the FF extracts have anti-oxidative and anti-inflammatory effects in vitro system, which can be used for developing pharmaceutical drug against oxidative stress and atherosclerosis.[This work was supported by the Dongguk University Research Fund and the MRC program of MOST/KOSEF (grant #: R13-2005-01001-0).]

Antibody fragments such as F(ab)’2 are advantageous over a whole antibody molecule in antigen-antibody binding studies because of its reduced non-specific binding, lower immunogenicity, and eliminated Fcassociated effector functions. Pepsin and/or papain are generally used to obtain Fab and F(ab)’2 [1]. But these enzymes have such disadvanages as long reaction time, acidic pH condition, and complicated purification. FabRICATOR, an IdeS, is a cysteine proteinase that can specifically cleave the hinge region of IgG at a neutral pH to generate well-defined Fc and F(ab)’2 fragments within 30 min [2]. It was purchased from Genovis, Inc., Sweden. In this study, we evaluated the FabRICATOR’s cleavage performance and devised the successive recovery process to obtain Fc and F(ab)’2. Rabbit IgG (gamma globulin) was used as a model IgG. For 50 μg of rabbit IgG, 100 unit of FabRICATOR was used at 37℃, pH 6.6 for 30 min. The isolation process consisted of three steps: (1) an immobilized metal affinity column to capture FabRICATOR by affinity interaction between the poly-histidine tag of the enzyme and the nickel domain, (2) Protein A column to recover Fc, and (3) SEC to obtain purified F(ab)’2. SDS-PAGE and SEC-HPLC (Biosep SEC-2000) results indicated the cleavage and separation were effective and successful. We plan to use F(ab)’2 for immunodiagnostics application replacing whole antibody and Fc for chemical fusion with other therapeutic protein for improved stability.

The pro-apoptotic tumor necrosis factor-related apoptosis inducing ligand (TRAIL) receptors, death receptor 4 (DR4) and DR5, are attractive targets to develop the receptor-specific agonistic monoclonal antibodies (mAbs) as anticancer agents because of their tumor-selective cell deathinducing activity(1-3). Here we report a novel agonistic mAb, AY4, raised against human DR4 in mice. ELISA analysis revealed that AY4 specifically bound to DR4 without competition with TRAIL for the binding. Despite the distinct binding regions of AY4 on DR4 from those of TRAIL, AY4 as a single agent induced caspase-dependent apoptotic cell death of several tumor types through the extrinsic and/or intrinsic pathways, without substantial cytotoxicity to normal human hepatocytes. Noticeably AY4 efficiently induced cell death of Jurkat cells, which were previously resistant to anti-DR4 agonistic mAbs, most likely due to the unique epitope property of AY4. AY4 also exhibited strong anti-tumor activity in xenograft model of human non-small cell lung carcinoma. Conclusively, our results provide further insight into the DR4-mediated cell death signaling and potential use of AY4 mAb as an anti-cancer therapeutics, particularly for DR4-responsive tumor types.

The aim of this work was to prepare liposome containing vancomycin and propolis for topical delivery.The vancomycin and propolis were successfully encapsulated in phosphatidyl choline-based liposomes and the possible improvement of their antioxidant and antimicrobial activities was tested against selected microbial. Particles with regular morphology, mean diameter ranging 100nm to 300nm, and good entrapment efficiency for propolis were obtained. The in vitro antimicrobial activity of nanoparticles was evaluated against microorganism of MRSA (Methicillin-resistant Staphylococcus aureus) and MRSE (Methicillin-resistant Staphylococcus epidermidis). The percentage of the encapsulated propolis and vancomycin was determined by uv-vis spectrophotometer. In order to investigate any possible synergistic or antagonistic effect between propolis and vancomycin, the antimicrobial activities of the mixtures, were also determined before and after encapsulation in liposomes. The antibiotics loaded liposomes can be used as a potential carrier for topical delivery.

Eicosapentaenoic acid (EPA) has been restricted to the use in food ingredients due to its unpleasant smells and rapid oxidation. In this study, EPA-contained nanoemulsions were encapsulated with maltodextrin to suppress the fishy odor and to inhibit a lipid oxidation. Their physicochemical properties were also investigated with deodorization rate. EPA-contained nanoemulsions were encapsulated by three methods such as, a freeze drying process (FDP), a spray- drying process (SDP), and a coacervation process (CP) with maltodextrin shell. The sizes of particles encapsulated from the EPA-contained nanoemulsions were ranged in 50-1,000 nm. The loading efficiencies of EPA in the nanoparticles prepared by FDP, SDP and CP were 15%, 42%, and 32%, respectively. They deodorized the fishy odor up to 90%. Peroxide values were also improved considerably compared to the raw EPA. The fishy odor of EPA was removed significantly by the encapsulation of EPA-contained nanoemulsions with maltodextrin. The encapsulation induced an enhancement of the antioxidant properties of EPA.

Topical natural antioxidants are a useful strategy for the prevention of oxidative stress mediated cardiovascular disease including atherosclerosis. From the viewpoint of this underlying principle, the screening of natural plant extracts with scavenging activity for pro-oxidant reactive species is a primary requirement for the development of new topical antioxidant formulations. PSORALEAE SEMEN (PS) is pharmaceutical name and it’s botanical name is Psoralea corylifolia Linne. This plant has been used as a traditional herbal medicine for the treatment of coronary artery disease, reducing blood flow and leucopenia. It has been reported that the extract of PS has pharmacological activities such as being anti-aging, anti-cancer and hematogenous effect. In the present study, extracts from PS were evaluated for their putative in vitro scavenging effects on reactive oxygen species (ROS) [superoxide anion, hydroxyl radical] and on reactive nitrogen species (RNS) [nitric oxide and peroxynitrite]. Moreover, we investigated the antioxidant activity by TEAC and DPPH assay and content of total phenolics as well as inhibitory effect on Cu2+-induced human LDL. The PS extracts presented a high potency to scavenge the tested ROS and RNS, as well as an inhibitory effect on LDL oxidation. In conclusion, the PS extracts have anti-oxidative and anti-atherosclerotic effects in vitro system, which can be used for developing pharmaceutical drug against oxidative stress and atherosclerosis. [This work was supported by the Dongguk University Research Fund and the MRC program of MOST/KOSEF (grant #: R13-2005-01001-0).]

The objective of the present work was to develop a new drug nanocarrier consisting of nanoparticles made of glycol chitosan(Gc) and eicosapentaenoic acid (EPA). We describe EPA conjugated Gc self-assembled nanoparticles as a promising system for inhibit inflammation in keratinocytes. Gc-EPA formed self-assembled nanoparticles spherical shape and diameter of approximately 100 nm by probe sonication in aqueous medium. EPA was loaded in to the Gc-EPA nanoparticles as a model drug. The chracteristics of EPA loaded Gc-EPA nanoparticles was analyzed using DLS, TEM and DSC. Their size increased from 250-450nm. The in vivo release behavior of EPA from Gc-EPA nanoparticles was studied by a dialysis method in phosphate buffered saline. EPA loaded Gc-EPA nanoparticles are valuable to enhance the absorption of a poorly water soluble water.

In present study, we isolated phloroglucinol derivative, 1-(3',5'-dihydroxyphenoxy)-7-(2'',4'',6-trihydroxyphenoxy)-2,4,9-trihydroxydibenzo-1,4-dioxin (1), from Ecklonia Cava and its potential inhibitory effect on adipocyte differentiation in 3T3-L1 cells. Lipid accumulation and expression of several genes associated with adipogenesis and lipolysis were examined at the end of differentiation period. Lipid accumulation was examined by measuring triglyceride contents and Oil-Red O staining. The expression levels of several genes and proteins were examined using reverse transcription-polymerase chain reaction (RT-PCR), real-time PCR and western blot analysis. Presence of compound 1 induced significant reduction of lipid accumulation and down-regulation of peroxisome proliferatoractivated receptor-g (PPARγ), sterol regulatory element-binding protein 1c (SREBP1c) and CCAAT/enhancer-binding proteins (C/EBPα) in a dose-dependent manner. Moreover, presence of compound 1 induced down-regulation of adipogenic target genes such as adipocyte fatty acid binding protein (aP2), fatty acid transport protein (FATP), fatty acid synthase (FAS), acyl-CoA synthetase 1 (ACS1), lipoprotein lipase (LPL) and leptin genes. According to the lipolytic response, compound 1 down-regulated perilipin and hormonesensitive lipase (HSL) and up-regulated tumor necrosis factor a (TNFa). Therefore, these results suggest that compound 1 might decrease lipid accumulation during adipocyte differentiation by modulation of adipogenesis and lipogenesis and compound 1 could be developed as a good functional agent to improve antiobesity.

Phloroglucinol derivatives, dioxinodehydroeckol (1) and 1-(3',5'-dihydroxyphenoxy)-7-(2'',4'',6-trihydroxyphenoxy)-2,4,9-trihydroxydibenzo-1,4-dioxin (2), were isolated from Ecklonia Cava. Their abilities to inhibit proliferation of MCF-7 and MDA-MB-231 human breast cancer cells were evaluated by measuring cell death via induction of apoptosis. The compound 1 exerted a higher anti-proliferative activity in the human breast cancer cells compared with compound 2. Compound 1 induced significant proliferative inhibition and apoptosis in a dosedependent manner in MCF-7 human cancer cells. Treatment with compound 1 induced the increase in caspase (-3 and -9) activities, DNA repair enzyme poly-(ADP-ribose) polymerase (PARP) cleavage, and pro-apoptotic gene and the decrease in anti-apoptotic gene. Besides, NF-κB family and -dependent activated genes were downregulated by compound 1. These results indicated that the potential inhibitory effect of compound 1 against growth of MCF-7 human breast cancer cells might be associated with induction of apoptosis through NF-κB or -dependent pathway. Although further studies are needed, the present results suggest that compound 1 has a promising potential as valuable chemopreventive agents.

Abilities of glucosamine and its derivatives (glucosamine-6-sulfate: SGlc-6, glucosamine-2-sulfate: SGlc-2 and glucosamine-6-phsphate: PGlc-6) to inhibit proliferation of MCF-7 human breast cancer cells were evaluated by measuring cell death via induction of apoptosis. Among them, glucosamine-2-sulfate (SGlc-2) exerted the highest antiproliferative activity in the human breast cancer. SGlc-2 induced significant proliferative inhibition and apoptosis in a dose- and timedependent manner in MCF-7 human cancer cells. Treatment with SGlc-2 induced the increase in caspase-3, -8 and -9 activities, DNA repair enzyme poly-(ADP-ribose) polymerase (PARP) cleavage and pro-apoptotic gene and the decrease in anti-apoptotic genes. Besides, NF-κB family and -dependent activated genes were down-regulated by SGlc-2. These results indicated that SGlc-2 had a potential against inhibition of growth of MCF-7 human breast cancer cells, which might be associated with induction of apoptosis through NF-κB or - dependent pathway.

Current tuberculosis (TB) treatment usually requires a long period to entirely remove bacteria within the body. Furthermore, active TB is generally treated with combinations of several antibiotics to reduce the risk of multi-drug resistance. It has been reported that essential oils are a good alternative to conventional TB antibiotics due to their excellent anti-TB activity and low toxicity within the body. Pulmonary delivery permits better exposure of drugs to the respiratory tissues and drugs can readily diffuse into the systemic circulation due to the presence of a large absorptive alveolar surface with a thin epithelial cell. In view of aerodynamics, it is regarded that the optimum size of particles should be 1~5 μm for efficient pulmonary delivery. In the present study, essential oils were encapsulated into poly(ethylene glycol) (PEG) particles using a supercritical particles from gas saturated solutions (PGSS) process. The effects of various operating conditions such as temperature, pressure, the content of essential oils and nozzle geometry on the morphology and encapsulation efficiency of essential oil loaded PEG particles were investigated in detail. Keywords : supercritical CO2 , PGSS , PEG , essential

Dioxinodehydroeckol was isolated from E. Cava and its inhibitory effect on lipid accumulation in cultured 3T3-L1 adipocytes were investigated by measuring TG contents and Oil-Red O staining and changes in genes and proteins associated with adipogenesis and lipolysis. Treatment with dioxinodehydroeckol significantly reduced lipid accumulation during adipocyte differentiation and induced down-regulation of SREBP1, PPARγ and C/EBPα in a dose-dependent manner. Moreover, dioxinodehydroeckol suppressed regulation of the adipocyte specific gene promoters such as FABP4, FATP1, FAS, LPL, ACS1 and leptin. As the lipolytic response, dioxinodehydroeckol down-regulated expression levels of perilipin and HSL genes and up-regulated the expression levels of TNFa mRNA compared to fully differentiated adipose tissue. Specific mechanism of dioxinodehydroeckol was examined through transcriptional downregulations of extracellular signal-regulated kinase (ERK). Therefore, these results suggest that dioxinodehydroeckol induced antiadipogenic effect on adipocytes via ERK-dependent signaling pathway.

Antioxidative and matrix metalloproteinase-2 and -9 (MMP-2 and -9) inhibitory effects of carboxymethyl-chitosan and -chitin (CM-chitosan and -chitin) were investigated in HT1080 cells. CM-chitosan and CM-chitin suppressed intracellular ROS formation by DCFH-DA and radical simulated oxidations of membrane protein and lipid in a concentrationdependent manner. In addition, a protective effect against oxidative damage of purified genomic DNA was observed in presence of CMchitosan and CM-chitin. Moreover, CM-chitosan and CM-chitin reduced the expression levels of MMP-2 and -9 in gelatin zymography, RT-PCR and western blot analysis without any cytotoxic influence. CM-chitin exhibited a higher MMP inhibitory effect than CM-chitosan through transcriptional down-regulations of AP-1 and NF-κB. Findings from the present study should underscore the nutraceutical value of CM-chitosan and CM-chitin as a potent anti-oxidant and MMP inhibitor via alleviations of radical-induced oxidative damage.

Recombinant protein is often produce by E.coli in the form of inclusion bodies (IBs). IBs are densed particle of proteins and do not have activity as protein. To convert IBs into water-soluble active protein, the refolding process is required. Ionic liquids (ILs) are organic salts composed of anion and cation, and the salts that do not crystallize at room temperature are called room temperature ionic liquids (RTILs) in particular. Tunable hydrophobicity and polarity of RTILs leads the expansion of its application in chemical and biological processes. In this study, we investigated how the hydrophobicity of RTILs affects protein refolding efficiency. Especially the effect of N-alkyl-N’-methylimidazolium methylsulfate having sulfur in anion part of ILs on target protein having the disulfide bond during refolding process. As the alkyl chain length increased the refolding efficiency generally decreased. In addition, we found that N-alkyl-N’- methylimidazolium methylsulfate is only effective as refolding additive when the target protein contain disulfide bond.

Ribose has existed D- and L- form isomers in nature. The use of Lcarbohydrates and their corresponding nucleotide in medicinal application has greatly increased. For example, L-ribose has been used as the starting material of curing hepatitis B in medicinal industry. However L-ribose, which is the enantiomer relationship with D-ribose, doesn't exist in nature. Therefore L-ribose has increased in demand as high-value products. Lribose was reported to be produced by enzyme, organic and epimerization reaction.L-ribose that is C-2 epimer relationship with L-arabinose can be obtained by epimerization of L-arabinose. In this study, the effects of the temperature, epimerization reaction and catalysts such as molybdenum(VI) oxide and molybdic acid were tested. Product concentrations of L-ribose was analyzed by Luna NH2 HPLC column. A comparative study on the yield of L-ribose was performed with change of reaction concentrations and catalysts.

Human BMP-7 has a process significant osteoinductive potential. BMP-7 mature form(amino acids 293-431)was inserted into the pET28a vector to produce the plasmid pBMP57. The E. coli strain BL21(DE3) was used as a host for rhBMP-7 production. The inclusion bodies are denatured and dialyzed into a refolding buffer. Several factors affecting the refolding yield, including refolding buffers have been investigated. The refolded BMP-7 is purified by FPLC on HiLoad 16/60 superdex column. SDS-PAGE assay and ELISA have been done to examine the concentration of refolded BMP-7. The aim of this study is to explore the in vitro refolding and purification protocol for BMP7 expressed in E.coli