Earticle

Over the last decades, significant efforts have been made to develop detection tools for monitoring hormones because of their biomedical importance. Well established detection methods for hormones are normally based on immunomeric assays. They, however, involve the use of labor-intensive and time consuming protocols. Here, we demonstrated a method to monitor the activity of human parathyroid hormone receptor (hPTHR), which belonging to B class G-protein coupled receptor (GPCR) family, using polypyrrole nanoparticle (PPyNP)-field effect transistor (FET) which allowed the development of a high-performance hormone sensor. In this work, PPyNP-FETs with uniform particle diameters of 20, 50 and 100 nm were functionalized with hPTHR produced from E. coli and detect target hormone peptide hormone, hPTH, with high sensitivity as low as picomolar concentration. In addition, hPTHR-PPyNP-FET showed high selectivity for target hormone detection and successfully recognized deletion form of hPTHs. This work may allow high-performance hormone sensor that can be used for the practical application including disease diagnosis and drug discovery. This method should also provide a powerful tool for basic research on GPCRs.

It has been reported that singe-strand (ss) DNA breaks could be photosensitized by some drugs. However, conventional methods to monitor the genotoxicity of the photosensitizing drugs are limited to the application for a high throughput screening of the photo-toxic drug because they are time consuming for the biological or chemical modification of the tested DNA and instrument-oriented. Therefore, We have investigated a colorimetric method to visualize ss DNA breaks using gold nanoparticles (AuNPs). ss DNA is known to physically adsorb onto AuNPs and prevent them from salt-induced aggregation. However, AuNPs mixed with UV irradiated ss DNA in the presence of photosensitizing drugs were aggregated after the salt addition into the mixture, resulting in red-to-purple color change. The extent of AuNP-aggregation was dependent on the UV-irradiation time and concentration of photosensitizers. In current, we are investigating the principle for the aggregation of AuNPs by only the ss DNA which were irradiated by UV light in the presence of ss DNA break photosensitizing drugs. The detail experimental results and working principle will be presented.

Food safety is a global health goal and the foodborne diseases take a major crisis on health. Therefore, detection of microbial pathogens in food is the solution to the prevention and recognition of problems related to health and safety. Conventional bacterial detection methods such as culture and colony counting methods may take up to a few days to yield an answer. Obviously this is inadequate, and recently many researchers are focusing towards the progress of rapid methods. Here, we demonstrate the rapid method which multiplex pathogen detection method based on capillary electrophoresis-single strand conformation polymorphism(CE-SSCP) coupled with multiplex ligation-dependent probe amplification(MLPA). Using ten foodborne pathogens as a model set, all the four MLPA-CE-SSCP steps were carefully optimized to precisely quantify by MLPA-CE-SSCP. We could obtain the results which illustrate a strong potential in clinical diagnosis, food safety, and biosafety.

In this study, liquid culture of Trichloma matsutake mycelia was carried out to biomass production and its inhibitory effects on melanin biosynthesis were evaluated. This Trichloma matsutake mycelia extract inhibited 38.6% of tyrosinase activity at 100 ppm, which is higher than that of extracelluar medium at same dose. In addition, when 100 ppm of Trichloma matsutake mycelia extract treated to melan-a cells for 3 days, 19% of melanin production was reduced without cell toxicity. These results suggested that cultured Trichloma matsutake mycelia might be useful for skin depigmenting material.

In this study the effects of 6,6’bieckol isolated from Ishige okamurae as a candidate for skin-whitening agent, we evaluated the inhibitory effect on mushroom tyrosinase and αMSH-induced melanin formation inhibitory effect in B16-F10 mouse melanoma cells.6,6’bieckol exhibited more potent tyrosinase inhibitory effect with IC50 value of 1.16μM than arbutin (IC50 = 243.16 μM) and kojic acid (IC50 = 40.28 μM), used as positive controls. Lineweaver-Burk plots suggest that 6,6’bieckol acts as a non-competitive inhibitors. Furthermore, these compounds were evaluated for their inhibitory effect on αMSH -induced melanin formation in B16-F10 mouse melanoma cells. Treatment with 6,6’bieckol (12.5-200 μM) resulted in a significant inhibition of melanin production in melanoma cells. Collectively, these results suggest that 6,6’bieckol might act as a skin-whitening agent via inhibition of tyrosinase activity and melanin formation in B16-F10 mouse melanoma cells.

The objective of this research was to investigate the whitening, anti-wrinkle, and anti-inflammatory effect of Jeju water containing vanadium component. In this study, Jeju water was divided into three groups (S1, S2, S3) according to its vanadium contents, i.e., the concentration of V for S1, S2, S3 were 8.0, 24.0, and 26.0 μg/l, respectively. Mushroom tyrosinase inhibition activity and anti-melanogenesis activity against melanoma cell were examined to explore Jeju water's whitening effects. In this test, Jeju water (S1, S2, S3) exhibited 6~8% inhibition activities on tyrosinase and 5~10% reduction of melanin contents in B16F10 melanoma cells. For the study of Anti-wrinkle effects, only S2 showed 5% inhibition on elastase acitivity. Jeju water (S1, S2, S3) showed 10~20% reduction on the lipopolysaccharide-induced secretion of nitric oxide in RAW264.7 cells, indicating its anti-inflammatory effects. In order to determine whether Jeju water could be safely applied to human skin, its cytotoxic effects were determined by colorimetric MTT assays in human dermal fibroblast. In this test, Jeju water did not show any cytotoxicity. All of these findings demonstrate that Jeju water containing vanadium component has beneficial effects which lead to improve human skin health.

Bioassay-guided investigation of the leaves of Neolitsea aciculata led to the isolation of 7 compounds: methyl linoleate (1), kaempferol 3-O-α-L- (2'',3''-E-di-p-coumaroyl)-rhamnoside (2), daucosterol (3), blumenol A (4), afzelin (5), quercitrin (6), 1,2-dilinolenoylglycerol-3-O- galactopyranosylglycerol (7). Their structures were elucidated on the basis of spectroscopic studies as well as by comparison with available data in the literature. Among these isolates, compond 6 showed significant DPPH free radical scavenging activity and SC50 was 21.7 ug/mL. Compounds 5, 6, and 7 showed significant elastase inhibition activities and IC50 was 56.8, 187.4, and 140.4 ug/mL, respectively.

We tried to analyze effects of Red Ginseng Oil (RGO) and Fractionated RGO (RGOf) on skin barrier function for both in vitro and in vivo. RGO was prepared by using supercritical extraction of the cake produced after extraction of red ginseng with boiling water. RGOf was prepared by hexane fractionation. The effect of RGO on the recovery of tape stripped skin barrier was measured by TEWL by using hairless mouse. Treatment of 0.1% RGO and RGOf showed the fastest recovery of skin barrier function expressed by TEWL compared to those of vehicle and 1% RGO. Alleviation effect of RGO on inflammation of human skin was also evaluated. RGOf significantly alleviated skin inflammation induced by UVB irradiation. In addition, the pro-inflammatory cytokines TNF- α was inhibited more than 60% in HaCaT keratinocytes by RGOf. And RGO and RGOf stimulated gene of differentiation markers, such as TG-1, Keratin 10 and involucrin. Collectively data shown in this study suggest that RGOf have strong skin protective activities by stimulating the epidermal homeostasis and preventing UVB induced skin inflammation.

Zanthoxylum is a genus of about 250 species of deciduous and evergreen trees and shrubs in the citrus or rue family, Rutaceae, native to warm temperature and subtropical areas worldwide. The genus Zanthoxylum has been credieted with a long list of ethnompharmacological properties. In the present study, in vitro anti-oxidant and anti-tyrosinase properties of extracts of Z. piperitum and Z. schinifolium leaves extracts were evaluated using various assays. Z. piperitum and Z. schinifolium leaves harvested at the mature green stage were collected at Baek-un Mt. Chollanam-do, S. Korea in August 2009. The extracts of Z. piperitum and Z. schinifolium leaves using methanol, n-hexane, chloroform, ethy acetate or butanol as solvents were evaluated for their tyrosinase inhibition, anti-oxidant and anti-microbial properties. The anti-tyrosinase and anti-oxidant potentials were determined by in vitro mushroom tyrosinase assay and the free radical scaening activity methods. Both of these results showed the strong inhibition abilities at a dosage of 100.0 ug/ml. Particularly, higher activity was exhibited by Z. piperitum with 92.8% and 80.1% inhibition of butanol and ethyl acetate extracts, while 78.2% and 57.8% inhibitions by Z. schinifolium respectively. However, both butanol extracts exhibited higher DPPH radical scavening activity than the corresponding methanol, n-hexane, chloroform, and ethyl acetate extracts. In addition, extracts of Z. piperitum leaves showed more potent anti-tyrosinase activity than Z. schinifolium leaves. Anti-microbial activities against Gram(+) and Gram(-) bacteria demonstrated good inhibition at 34.8 –43.6 ug/ml. These results obtained from biological assays showed that extracts of Z. piperitum and Z. schinifolium leaves possessed multiple bioactivities, including anti-tyrosinase, anti-oxidants, anti-microorganism and cell proliferation. The data exhibited the high potential of applying extracts of Z. piperitum and Z. schinifolium leaves in cosmoceutical lines.

Indole-3-acetic acid (IAA) is known as a plant growth hormone. Studies were made to evaluate the anti-wrinkle effects of IAA by comparing the changes of proteins related to collagen synthesis in human dermal fi broblast cells by using two-dimension electrophoresis. Cellular cytotoxicity was measured according to a rapid colorimetric MTT assay and the antioxidant activity was also observed by using DPPH methodology. Results showed that IAA was not toxic to fibroblast at the concentration range tested. Proteome profiling analysis showed that IAA promoted the expression of proteins related to collagen production. IAA could be considered as an attractive, anti-wrinkle agent for cosmetic applications.

(-)-α-bisabolol is a sesquiterpene alcohol found in the oils of chamomile and other plants. (-)-α-bisabolol has been widely used in dermatological and cosmetic formulations as an anti-inflammatory agent. But so far there were no studies that show the inhibitory mechanism of (-)-α-bisabolol in inflammation-associated gene expression. Therefore, this study was designed to investigate the anti-inflammatory effect of (-)-α-bisabolol and its mechanisms of action. Among many pro-inflammatory mediators, we found that (-)-α-bisabolol inhibited LPS-induced production of nitric oxide (NO) and prostaglandin E2 (PGE2) in RAW264.7 cells. In addition, in iNOS and COX-2 promoter luciferase assays, (-)-α-bisabolol reduced LPS-induced activation of iNOS and COX-2 promoters. Protein levels of iNOS and COX-2 were also reduced by (-)-α-bisabolol in a concentration-dependent manner. These results suggest that (-)-α-bisabolol exerts anti-inflammatory effects by downregulating expression of iNOS and COX-2.In order to elucidate inhibitory mechanisms of (-)-α-bisabolol on expression of iNOS and COX-2, we investigated effects of (-)-α-bisabolol on LPS-induced activation of AP-1 and NF-kB elements which exist in the promoter of iNOS and COX-2 genes. LPS-induced activation of AP-1 and NF-kB promoters was significantly reduced by (-)-α-bisabolol, suggesting that AP-1 and NF-kB promoters are involved in (-)-α-bisabolol effects. To further confirm this, ELISA for phospho-IkBa and Western blot for phospho-p42/44mapk, phospho-p38mapk, and phospho-JNK were performed. (-)-α-bisabolol reduced LPS-induced phosphorylation of IkBa. In addition, while LPS-induced phosphorylation of p42/44mapk and p38mapk was attenuated by (-)-α-bisabolol, significant change in the level of phosphorylated JNK was not observed. Collectively, our results indicate that (-)-α-bisabolol exerts anti-inflammatory effects by downregulating expression of iNOS and COX-2 genes through the inhibition of NF-kB and AP-1 (p42/44mapk and p38mapk) signaling pathway and suggest that (-)-α-bisabolol may be introduced for the treatment of inflammatory diseases.

Nigella glandulifera (Ranunculaceae) has been used as a folk remedy for treatment of colds, cough and insomnia. The seeds are believed to have diuretic, analgesic, spasmolytic, galactagogue, bronchodilator properties, and to cure edema, urinary calculus, and bronchial asthma. To develop a novel skin depigmenting agent from natural sources, the inhibition of melanogenesis by N. glandulifrea, was evaluated. The methanol extracts of this plant showed significant down-regulated melanin synthesis in a dose dependent manner at a non-toxic concentration in cultured B16F10 mouse melanoma cells. This extract was further fractionated by using solvent-solvent partition and silica open column chromatography to identify the active components. From GC-MS data, oleic acid methyl ester was found as one of the depigmenting agents. In conclusion, we suggest that this fraction may be a safe and effective depigmentation agent.

Torreya nucifera S. et Z. (Torreya) is a slow-growing coniferous tree native to South Korea's Jeju Island and to southern Japan. Its leaves are evergreen, needle-like, 2～3 cm long and 3 mm broad, with a sharply spined tip and two whitish stomatal bands on the underside. The torreya oil was used for edible, hair, lighting, and medical purposes. Recently, the usage of this oil is drawing attention in the cosmetics and fragrance industry. Thus this study focused on extraction of Jeju torreya using supercritical carbon dioxide (SCO2). The leaves of Jeju torreya were sampled (3 kg), air-dried, and chopped. Then, the essential oil was extracted using SCO2, maintaining a pressure and a temperature as 350 bar and 60 ℃, respectively. Analysis of the chemical composition of essential oil from Jeju torreya leaves was carried out using gas chromatography mass (GC/MS). As a result, the optimum yield of SCO2 extraction was found as 1.5 %. And more than 80% of the total oil components were identified, with Cyclotetrasiloxane, Dehydroabietic acid and Pyrido[2,3-b]indole as the main compounds in essential oil from Jeju torreya leaves. These results indicate that SCO2 extraction can be developed to be an eco-friendly process for obtaining the valuable essential oils such as Jeju torreya oil.

Today Mongolia is one of the countries which officially leads its system of herbal medicine. According to their own culture and beliefs, Mongolian has developed their system of medicine, and become famous. As Chrysanthemum zawadskii, Mentha arvensis, and Lophantus chinensis are used for typical medicinal plant, this study was aimed to exploit extracts of those plants for developing functional cosmetic applications. Each sample was prepared by 0.0005%, 0.001%, and 0.002% of ethanol extraction respectively. The effect of extracts on expression of skin differentiation m arkers such as filaggrin and involucrin was measured by RT-PCR with HaCaT keratinocytes. This study showed that those differentiation markers were expressed in a dose dependent manner comparing with Ca2+ as a positive control. In addition, to determine the effect of mongolian medicinal plants on the inflammation, the pro-inflammatory cytokine, TNF-α expression in HaCaT keratinocyte will be examined by ELISA. Therefore, Mongolian medicinal plant; Chrysanthemum zawadskii, Mentha arvensis, and Lophantus chinensis extracts have affirmative effects on skin differentiation.

Sep-Pak C18 cartridges were investigated to evaluate their ability to purify anti-photoaging compound extracted from Ficus deltoidea (FD) leave. This cartridge is enable efficient purification of peptides and small proteins before analytical techniques. Several studies have been reported that Ficus species contain polypeptide compounds that known to have strong antioxidant properties. In our previous data FD water extract shows high anti-photoaging and anti-melanogenic activity. From this observation it might be a useful idea to purify the peptide compound from FD extract that responsible to protect UVB-induced skin cells from photoaging. Therefore, in the present study, we investigate the effect of cleanup FD extract on the expression of TNF-α and MMP-1, and collagen synthesis in UVB irradiated cultured human fibroblast and keratinocytes. FD extract were diluted with 1% trifluoroacetic acid (TFA), 0.1 M Amm. Bicarb (ABC) and distilled water separately, followed by filtration and dried using freeze dryer, prior to the Sep-Pak C18 cleanup. These buffers represent the different properties of peptide eluted from the cartridges. The treatment of all samples inhibited UVB-induced TNF-α expression. Distilled water is the preferred diluents that reduce TNF-α expression induced by UVB irradiation more than 70% at 0.005% compare to dexamethason. The use of sample diluents such as distilled water and ABC were reduced UVB-induced MMP-1 expression in a dose-dependent manner. The treatment of fibroblast with both samples were also increased the reduction of UVB-induced collagen synthesis. Therefore, these results point to the potential use of Sep-Pak C18 to cleanup FD extract for purifying the anti-photoaging compound.

Solvent fractions of apple flower leaf was investigated for the screening of functional food and cosmetic materials. The content of phenolic compound and flavonoid in ethylacetate fraction was measured as 809.90 ㎎/g and 140 ㎎/g. The DPPH radical scavenging activities was over 80% at a concentration of 200 ㎍�/㎕and SOD like activity was over 90% at 50 ㎍�/㎖concentration in ethylacetate fraction. Tyrosinase inhibition activity related to skin-whitening was over 60% by ethylacetate fraction at 100 ㎍�/㎖�. As an anti-aging effect, the elastase inhibitory activity was about 45% in ethylacetate fraction. The highly active compounds were isolated and identified as kaempferol by GC/MSD analysis. In conclusion, it was indicated that apple flower leaf extract could be used for dietary and cosmetical purpose.

Vitiligo is an acquired disorder of pigmentation in which depigmentation of skin and hairs occur due to the loss of functional melanocytes from the epidermis. It is known that the existence of inactive melanoblasts in the hair root follicles provides a melanocyte source for repigmentation in vitiligo. Migration of these melanocyte precursors from the outer root sheath of hair follicles into clinically depigmented epidermis is crucial for repigmentation in vitiligo therapy. Although the current natural compounds may induce varying degrees of repigmentation in vitiligo lesions, treatment challenges persist, as either not all patients respond to available therapies, and the treatment duration is too long or sometimes a plateau effect is seen with the treatment becoming less effective after an initial response. To evaluate the potential of herbal candidates in treatment of vitiligo, the effect of methanol extracts of Longanae Arillus (MELA) on differentiation, proliferation and migration in melanoblast cell was done. The results showed that, MELA could induce the differentiation and migration in melanoblast cell. From proliferation assay, MELA did not exhibit the cell toxicity to the melanoblast cell. The findings of the present study may be important in developing safer strategies for vitiligo treatment.

Extracts of Mongolian herbal plants such as serratula centauroides, comarum zalesovianum, sedum hybridum, ziziphora pamiralaica, lonicera mycrophylla, chrysanthemum zawadskii, mentha arvensis, lophantus chinensis, orastachys fimbrinata and dracocephalum grandiflora have been widely used as traditional medical applications in Mongolia. Since some of these plant extracts have been shown to have strong anti-oxidant activities, anti-melanogenesis effect of these extracts were analyzed. In order to perform melanin analysis at non-toxic concentrations, cytotoxic study of each extracts were first measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Out of these ten plant extracts, orostachys fimbrinata extract showed the strongest anti-melanogenesis activity. In this study, the ethanolic extract from orostachys fimbrinata was assessed with regard to its effects on melanogenesis in B16F1 melanoma cells. The orostachys fimbrinata extract was shown to inhibit melanin synthesis and tyrosinase activity in a dose-dependent manner without any significant effects on cells proliferation. In the range of non-cytotoxic concentrations at 0.0125~0.01% w/v, formation of melanin from cultured B16F1 melanoma induced by α-melanocyte stimulating hormone (α-MSH) significantly inhibited by treatment of orostachys fimbrinata extract. By using spectrophotometer, the results obtained clearly shows that 0.01% w/v orostachys fimbrinata extract, inhibit α-MSH induced melanogenesis almost 3-folds, which determine orostachys fimbrinata extract has potential to be used as a novel depigmenting agent for cosmetics.

MITF, an E-Box (CATGTG) binding transcriptional factor, regulates expression of tyrosinase, which involves in melanin synthesis in the Melanocytes. To screen for MITF and E-box binding inhibitor, EMSA (Electrophoretic Mobility Shift Assay) was performed. Based on the computer-simulated structure, 27 chemicals were selected and were investigated as MITF and E-box binding inhibitors. After that, cell tests were performed by using the mean-a cell line. Among them, compound #18 was shoed the potent inhibitory activity against MITF and E-box binding. Pursuant to EMSA, intensities of MITF and E-box bands were reduced by compound #18 in a dose-dependent manner. In addition, unlabelled Tyr-p, mutated Tyr-p and random oligomer were used for EMSA to investigate the competitive binding inhibitory effect. As a result, excess unlabelled Tyr-p was an effective binding competitor. To confirm the above findings, the expression of tyrosinase was evaluated by western blot, in response to compound #18. It was found to be reduces level tyrosinase while MITF and TRP-1 remain unaffected. Therefore, compound #18 could be used as a specific MITF and E-box binding inhibitor, to reduce the melanin content in Melanocytes.

Studies were made to develop a new skin whitening agent with Cirsium setosum (Wild.) MB. Various solvents were used to extract natural products from Cirsium setosum (Wild.) MB. The free radical scavenging activities of extracts were tested by DPPH assays. Enzymatic inhibitory melanogenesis was tested by mushroom tyrosinase inhibition effects of extracts. Melanin content in the B16 cell was also measured to observe direct skin whitening effects of extracts. In addition to inhibitory melanogenesis effects, the low cytotoxicity by MTT assay showed that Cirsium setosum (Wild.) MB had a high potential to develop a new natural whitening agent for cosmetics.

In the present study, effect of 7′-phloroeckol isolated from Ecklonia cava on the growth of human epidermal keratinocyte growth was investigated by measuring cell proliferative and anti-apoptotic activities in HaCaT cells. The cell proliferative and anti-apoptotic activities were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and Western blotting. Treatment with the 7′- phloroeckol increased the proliferation of HaCaT cells by 259.2% at a concentration of 0.5 μM compare with control. The protein expression levels of phosphorylated-extracellular signal-regulated kinases (p-Erk) and p-Akt were significant increased by 7′-phloroeckol. Apoptosis related Bcl-2/Bax ratio was also increased in a dose dependent manner. Consequently, 7′-phloroeckol has the potential to promote the human epidermal keratinocyte through stimulating proliferation and preventing apoptosis.

Citrus fruits contain 35~55% juice and rest as solid biomass, only 5% of this biomass utilizes as silage and rest are treated as waste material. The management of the waste is very expensive and non productive. Thus we have focused to produce GABA and antimicrobials by fermentation using lactic acid bacteria. The present aim is to develop an effective and simple process to convert the citrus waste into value added commodity. The citrus silage of one to three years maturity level was collected and evaluated the bio-organic properties. For screening of production of antimicrobials, we have used seven strains from KCCM and 37 strains isolated from porcine feces. The culture supernatant was used to evaluate the antimicrobial activity against E. coli O-157, S. aureus, E. coli K88, and Salmonella spp. S. gallinarum, S. enteritidis, and S. typhimurium. Out of all screened bacteria 23 strains were found to be antagonistic for S. enteritidis and S. typhimurium. The three strains namely L. reuteri LA17, B. subtilis 3-3, L. brevis IFO 12005 were secreted antimicrobials for S. enteritidis and S. typhimurium. In addition to that Factorial Design was used to optimize the GABA production from same silage, at different nitrogen source.

Fucoidans are sulphated polysaccharides having L-fucose as major constituent. For the past decade fucoidans isolated from various species have been extensively studied due to their numerous biological activities, including antitumor and immunomodulatory, antivirus, anticoagulant and antithrombotic, anti-inflammatory, antioxidant and and many more. However, the effect of different sizes of fucoidan on melanin synthesis has not been elucidated yet. Therefore, the aim of the present study is to identify the skin whitening effect of the fucoidans in respect to molecular weights. To confirm the depigmenting effect of the native and the acid hydrolyzed fuicoidan fractions, from Fucus vesiculosus were subjected to B16F10 melanoma and melan-a melanocyte cell line. The Cell viability (MTT) and melanin production were evaluated to elucidate the effect of fucoidan on depigmentation. The results showed that the crude fucoidan isolated from Fucus vesiculosus exhibited the melanin inhibition effect without cell toxicity up to the concentration 100 ug/ml in both cells. The purification of the hydrolyzed fractions required prior to establish the size effect of fucoidan on depigmentation. We anticipate the use of the fucoidan in different Cosmeceutical as depigmenting agent.

Bioassay-guided investigation of the stem of Cleyera japonica Thunb. led to the isolation of five compounds such as 3,5,7-trihydroxylchromone 3-O-α-L-rhamnopyranoside (1), aviculin (2), 3,3'-di-O-methylellagic acid (3), 3,3'-di-O-methylellagic acid 4'-O-β-D-xylopyranoside (4) and betulinic acid (5). Their structures were elucidated on the basis of spectral studies as well as by comparison of their data with literature values. In order to study the skin related bioactivities for the isolated compounds, screenings on anti-oxidation, anti-tyrosinase and anti-elastase were conducted. For the anti-oxidation tests, compound 1, 2 and 3 showed strong DPPH radical scavenging activities with SC50 of 59.3, 58.1 and 123.3 μg/mL respectively, whose activities were comparable to a positive control BHT (SC50 89.0 μg/mL). On the tyrosinase inhibition studies, compound 4 (IC50 36.3 μg/mL) showed more potent activity than arbutin (IC50 67.2 μg/mL). On the elastase inhibition studies, compound 5 (IC50 17.9 μg/mL) showed higher activity than oleanolic acid (IC50 29.6 μg/mL), a positive control. Based on these results, C. japonica stem extracts could be potentially applicable as an ingredient in skin care formulations.

Megestrol acetate (MGA) is an appetite stimulant indicated for cachexia in patients with AIDS or cancer [1]. MGA is practically insoluble in water. The commercial formulation is available in microcrystal dispersion which shows extremely low oral absorption and large variation by food intake [2]. We designed new nano-formulation of megestrol acetate to enhance an oral bioavailability of megestrol acetate and studied the pharmacokinetic properties of our formulation under fed and fasting conditions.In this study, we designed nanoemulsions for the enhanced oral bioavailability of MGA. Oil, surfactant and cosurfactant were selected base on the solubility study. The mean particle size of nanoemulsions was estimated on zeta-sizer (ZS90, Malvern, England). Dissolution of MGA was estimated on dissolution tester (U.S. Pharmacopeia, Dissolution test, Apparatus 2). Relative oral bioavailability of our nano-formulation of MGA was compared with the commercial product (Megace® oral suspension, Bristol-Myers Squibb Co.) against dogs on fed and tasted state following cross-over design. The MGA nanoemulsions have 25nm of droplet size and over 80 % of dissolution rate. Relative oral bioavailability of MGA nanoemulsions compared to commercial oral suspension was 450 %, 150% in fasted and fed state, respectively.New nanoemulsions formulation of MGA was successfully designed and showed a good feasibility for the further clinical study. The enhanced oral bioavailability of MGA could assure the more reliable cancer chemotherapy in future.

In this study, eight kinds of phenolic acid conjugated COSs were synthesized from benzoic acid and cinnamic acid. Eight kinds of chitooligosaccharides (COS) with different substitution groups, including p-hydroxyl (HBA-COS, PCA-COS), 3,4-dihydroxyl (PTA-COS, CFA-COS), 3-methoxyl-4-hydroxyl (VNA-COS, FRA-COS) and 3,5-dimethoxyl- 4-hydroxy (SRA-COS, SNA-COS) groups, were evaluated for their inhibitory activities against β-Secretase (BACE). Cinnamic acid derivatives showed the strongest BACE inhibitory activity more than benzoic acid derivatives. In addition, catechol structure substantially increased the BACE inhibitory activity of the COS derivatives. CFA-COS was further analyzed for its inhibitory pattern on BACE via Lineweaver–urk plots. CFA-COS showed non-competitive inhibition of BACE. In this study, we suggested that phenolic acid conjugated COS derivatives have potential as BACE inhibitors for preventing Alzheimer’ disease.

In this study, the ability of phenolic acid conjugated of chitooligosaccharides (COSs) were evaluated for their inhibitory effect on proliferation of B16-F10 mouse melanoma cells. Phenolic acid conjugated COSs-induced cell death was characterized by cell viability assay. According to our results, all protocatechuic acid conjugated-COS (PTA-COS) significantly induced cell death in B16-F10 cells. Apoptosis was determined by cell morphology and electrophoresis of DNA fragmentations. Furthermore, treatment with PTA-COS also induced the increase in caspase activity, PARP cleavage, pro-apoptotic proteins (Apaf-1,Bax, Bad, Fas, and p53) and the decrease in anti-apoptotic proteins (Bcl-2, Bcl-xL and XIAP). In addition, phospho-CREB and phospho-Akt were down-regulated by PTA-COS. These results indicated that the potential inhibitory effect of PTA-COS against growth of B16-F10 cells which might be associated with induction of apoptosis through Akt dependent p53 and CREB pathways.

HER2 is over-expressed on the surface of breast cancer cells and involved in the development of cancer tumors by uncontrollable phosphorylation signalling at the early stage of MAPK pathway. Herceptin treatment inhibits such excessive kinase activation upon binding to the three regions of extracellular domain of HER2, thereby disrupting the signal production. However, the cost, the potential side effects and the increased herceptin resistance act as drawbacks of herceptin treatment. Therefore, an alternative therapy for breast cancer has been required. Inspired by the understanding of herceptin interaction with HER2 epitope, we screened DNA aptamers cognitive of a peptide constituting one of the three HER2 epitopes with the use of a monolith-based IMAC SELEX method. This method enables multiple equilibrium stages, high resolution and real-time monitoring during the screening process, thereby greatly increasing the efficiency of hitting HER2 affinity aptamers. Potential applications of HER2 specific aptamers will include hyperthermia, targeted drug delivery, magnetic resonance imaging, detection of blood-level biomarker and so on.

Many marine bacteria have been found to produce a large amount of alginate lyases and many genes encoding alginate lyases from marine bacteria have been cloned and sequenced. Alginate lyase mutants with high activity against alginate are highly desired for use in the food, pharmaceutical, and medical industries. Hence, we have amplified the alyVI gene from the marine bacterium Vibrio sp. QY101 using plasmid pGEX-4T-1 for mutagenesis. We have also performed a computational study of alginate binding with wild-type and mutants of alyVI. Our goal is to increase the catalytic efficiency of alyVI toward alginate by site-directed mutagenesis based on computational technology. The combined molecular docking, molecular dynamics simulations, binding free energy calculations, and binding energy decompositions provide valuable insights into the detailed binding of Aliginate Lyase AlyVI with its substrate. Subsequently the computational simulations followed by site directed mutagenesis and AlyVI activity assays, have led to a detailed understanding of Aliginate Lyase AlyVI interacting with its substrate. Results obtained from the binding energy decomposition reveal the contribution of each residue at the protein-ligand interaction interface to the binding affinity. The data from wet experimental tests are consistent with the computational predictions.

Abstract The purpose of this study was to enhance cellular accumulation of drugs and therapeutic effects in cells in the local nano-delivery system, leading to effective killing cells. To synthesize biodegradable-cationic polymer, hydroxyl groups of the polymer were activated by carbonyldiimidazole, and reacted with low molecular cationic polymer. The cationic polymer synthesis was verified by 1H-NMR. In 1HNMR spectra ofbiodegradable - cationic polymers, proton signals of the cationic polymer were confirmed. Therapeutic agent loaded biodegradable-cationic nanoparticles ware prepared using a modified sonication method. The mean diameter of cationic nanoparticles was analyzed by using a Zetasizer. The biodegradable-cationic nanoparticels have mono-disperse size distribution in the range from 100 to 500 nm. The cytotoxicity was measured by an MTT assay. An ability to kill cancer cells, treated with drug-loaded bio-degradable cationic nanoparticles, was significantly higher than that with the free drug. Conclusions The biodegradable cationic nanoparticles were synthesized by carbonyldiimidazole method. Therapeutic agent loaded cationic nanoparticle showed enhanced cytotoxicity than a similar amount of unloaded nanoparticle and free drug. These findings suggest that cationic nanoparticle may be a viable alternative for development of chemotherapeutic carreiers.