Development of a Simple Direct and Hot-Start PCR Using Escherichia coli-Expressing Taq DNA Polymerase.

Taq DNA polymerases have played an important role in molecular biology for several years and are frequently used for polymerase chain reaction (PCR); hence, there is an increasing interest in developing a convenient method for preparing Taq DNA polymerase for routine use in laboratories. We developed a method using Escherichia coli (E. coli) that expresses thermostable Taq DNA polymerase directly in the PCR without purification. The Taq gene was transformed into E. coli and expressed. After overnight incubation and washing, E. coli-expressing Taq DNA polymerase (EcoliTaq) was used as the DNA polymerase without purification. EcoliTaq showed activity comparable to that of commercial DNA polymerase and remained stable for 3 months. With a high-pH buffer containing 2% Tween 20 and 0.4 M trehalose, EcoliTaq facilitated direct PCR amplification from anticoagulated whole blood samples. EcoliTaq exhibited good performance in allele-specific PCR using both purified DNA and whole blood samples. Furthermore, it proved to be useful as a DNA polymerase in hot-start PCR by effectively minimizing non-specific amplification. We developed a simple and cost-effective direct and hot-start PCR method in which EcoliTaq was used directly as a PCR enzyme, thus eliminating the laborious and time-consuming steps of polymerase purification.

Synergistic Effect of Saccharin and Caffeine on Antiproliferative Activity in Human Ovarian Carcinoma Ovcar-3 Cells.

The purpose of this study was to confirm the antiproliferative and apoptotic induction potential of a saccharin and caffeine combination in ovarian cancer cells. The cell line used was Ovcar-3, and the cell viability was measured through a WST-8 assay, while a Chou-Talalay assay was used to confirm the synergistic effect of saccharin and caffeine on the ovarian cancer cells. A clonogenic assay, annexin V-FITC/PI-PE double-staining, and RT-PCR were performed to confirm the expression of genes that induce colony formation, cell viability, and apoptosis in ovarian cancer cells treated with the saccharin-caffeine combination. It was demonstrated that both saccharin and caffeine decreased the viability of Ovcar-3 cells, and the cell viability decreased even more significantly when the cells were treated with the combination of saccharin and caffeine. The clonogenic assay results showed that the number of colonies decreased the most when saccharin and caffeine were combined, and the number of colonies also significantly decreased compared to the single-treatment groups. Based on flow cytometry analysis using annexin V-FITC/PI-PE double-staining, it was confirmed that the decrease in cell viability caused by the combination of saccharin and caffeine was correlated with the induction of apoptosis. The results of the RT-PCR confirmed that the combined treatment of saccharin and caffeine promoted cell apoptosis by regulating the expression of apoptosis-inducing genes. These results demonstrate that the combination of saccharin and caffeine more efficiently inhibits the proliferation of Ovcar-3 cells and induces apoptosis in vitro.

Krill Oil Inhibits Cholesterol Synthesis and Stimulated Cholesterol Excretion in Hypercholesterolemic Rats.

The present study aimed to investigate the antihypercholesterolemic effects of krill oil supplementation in high-cholesterol diet-induced hypercholesterolemic rats, and the mechanisms underlying these effects. Rats were divided into five groups: normal control, control (high-cholesterol diet), krill oil 100 mg/kg b.w. (high-cholesterol diet with Krill oil 100 mg/kg b.w.), and krill oil 200 mg/kg b.w. (high-cholesterol diet with Krill oil 200 mg/kg b.w.). After 12 weeks, the rats were sacrificed to observe the effects of krill oil on cholesterol synthesis and excretion. We found that krill oil supplementation suppressed total triglycerides, total cholesterol, and LDL-cholesterol levels, as well as HMG-CoA reductase activity. It stimulated AMPK phosphorylation, LDL receptor and ACAT2 expression in the liver, and the fecal output of cholesterol. Furthermore, it decreased the levels of P-selectin, sVCAM-1, and NO, as well as aortic wall thickness, demonstrating its role in the prevention of atherosclerosis. Thus, we suggest that krill oil supplementation can reduce LDL-cholesterol levels in the blood during hypercholesterolemia by stimulating the uptake of LDL-cholesterol into tissue and cholesterol excretion, as well as inhibition of cholesterol synthesis.

Chitinophaga aurantiaca sp. nov., isolated from a soil sample from a tangerine field.

A Gram-stain-negative, facultatively anaerobic, non-motile and rod-shaped bacterial strain, designated THG-SD5.5T, was isolated from a soil sample collected in a tangerine field, Republic of Korea. According to the 16S rRNA gene sequence comparisons, the isolate was identified as a member of the genus Chitinophaga and to be closely related to Chitinophaga ginsengihumi KACC 17604T (97.9%) and Chitinophaga rupis KACC 14521T (97.5%). The 16S rRNA gene sequence similarities with other species of the genus Chitinophaga were in the range 92.8-95.5%. Catalase test was positive. Oxidase test was negative. The DNA G + C content was determined to be 46.1 mol%. DNA-DNA hybridization values between strain THG-SD5.5T and C. ginsengihumi KACC 17604T and C. rupis KACC 14521T were 45.1% and 15.6%, respectively. Strain THG-SD5.5T was also found to be able to grow at 24-33 °C, at 0-5% NaCl and at pH 5.5-9.0. The major fatty acids were identified as anteiso-C15:0, C16:0, anteiso-C17:0 and C18:0. The dominant respiratory quinone was identified as menaquinone-7 (MK-7). The polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, an unidentified aminolipid, two unidentified phospholipids and three unidentified lipids. Based on these phenotypic, genotypic and phylogenetic characterisations, strain THG-SD5.5T (= KACC 19338T = CGMCC 1.16304T) is concluded to represent a novel species of the genus Chitinophaga, for which the name Chitinophaga aurantiaca sp. nov. is proposed.

Cellulomonas aurantiaca sp. nov., isolated from a soil sample from a tangerine field.

A Gram-stain positive, facultatively aerobic, motile and rod-shaped bacterial strain, designated THG-SMD2.3T, was isolated from a soil sample collected in a tangerine field, Republic of Korea. According to the 16S rRNA gene sequence comparisons, the isolate was identified as a member of the genus Cellulomonas and to be closely related to Cellulomonas fimi ATCC 484T (98.5%), Cellulomonas biazotea DSM 20112T (98.3%), Cellulomonas chitinilytica X.bu-bT (98.0%), Cellulomonas xylanilytica XIL11T (97.2%), Cellulomonas humilata ATCC 25174T (97.1%) and Cellulomonas composti TR7-06T (97.0%). The 16S rRNA gene sequence similarities with other current species of the genus Cellulomonas were in the range 95.4-96.6%. Catalase and oxidase tests were found to be positive. The DNA G+C content was determined to be 73.0 mol%. DNA-DNA hybridization values between strain THG-SMD2.3T and C. fimi ATCC 484T, C. biazotea DSM 20112T, C. chitinilytica X.bu-bT, C. xylanilytica XIL11T, C. humilata ATCC 25174T and C. composti TR7-06T were 58.1 ± 1.6%, 56.7 ± 0.8%, 30.3 ± 1.6%, 22.8 ± 1.6%, 19.9 ± 1.6%, and 13.5 ± 3.0%, respectively. Strain THG-SMD2.3T was also found to be able to grow at 20-42 °C, at 0-3% NaCl and at pH 5.5-10. The major fatty acids were identified as anteiso-C15:0, iso-C15:0, anteiso-C17:0 and iso-C14:0. The predominant menaquinone was identified as tetrahydrogenated menaquinones with nine isoprene units [MK-9(H4)]. The polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unidentified aminolipids and two unidentified phospholipids. Based on these phenotypic, genotypic and phylogenetic characterisations strain THG-SMD2.3T (= KACC 19341T = CGMCC 1.16303T) is concluded to represent a novel species of the genus Cellulomonas, for which the name Cellulomonas aurantiaca sp. nov. is proposed.

A pilot randomized controlled trial of 6-week combined exercise program on fasting insulin and fitness levels in individuals with spinal cord injury.

PURPOSE: The aim of this randomized controlled trial study was to investigate the effect of combined exercise program on the fasting insulin and fitness levels of people with spinal cord injury (SCI). METHODS: A total of 19 individuals with SCI participated in a combined exercise program consisting of aerobic and resistance exercises for 60 min per day, 3 days per week for 6 weeks. Peak oxygen consumption, body mass index, percent body fat, waist circumference, shoulder abduction and adduction, shoulder flexion and extension, elbow flexion and extension, fasting insulin levels, and homeostasis model assessment of insulin resistance (HOMA-IR) levels were measured at baseline and after the intervention. RESULTS: The 6-week exercise program significantly decreased the average fasting insulin (baseline: 7.5 ± 4.7 µU/ml vs. post-intervention: 4.5 ± 2.2 µU/ml, p < 0.05) and HOMA-IR (baseline: 1.5 ± 1.0 vs. post-intervention: 0.9 ± 0.4, p < 0.05) in the exercise group, whereas there was no change in control group (between group difference, mean fasting insulin: - 3.2 µU/ml, p = 0.003; mean HOMA-IR: - 0.66, p = 0.001). In addition, muscle strength of the shoulder flexors, extensors, abductors, adductors, and elbow flexors was significantly improved in the exercise group compared to the controls. CONCLUSION: A combined exercise program is effective in decreasing fasting insulin and HOMA-IR levels while improving fitness in those with SCI. These slides can be retrieved under Electronic Supplementary Material.

Characterization of ZnS Thin Films Grown Using Chemical Bath Deposition with Three Different Complexing Agents.

Cadmium sulfide buffer layer was replaced with zinc sulfide thin film owing to toxicity. ZnS thin films were fabricated using chemical bath deposition. The inhibition of Zn(OH)2 and high uniformity are important factors for the deposition of ZnS. The characteristics of ZnS thin films were analyzed by adding ethylenediamine tetra-acetate acid (EDTA) and hexamethylene tetramine (HMTA). The morphological characteristics of the ZnS buffer layer are closely related to the use of a complexing agent which controls the concentration of Zn2+ ions and Zn(OH)2 in the deposition process. The addition of the complexing agent EDTA accelerated the cluster-cluster method but it exhibited lower uniformity and greater cracking phenomenon. HMTA can be effectively applied to increase the amount of Zn2+ ions forming ZnS. It can be easily found as Zn2 HMTA at high temperatures. The results of the experiment with the addition of HMTA revealed that the surface of the thin film did not change with the increase in the concentration of HMTA, but the thickness of the thin film increased gradually. HMTA promoted the ion-ion method to grow the thin film uniformly, but the speed was slow. Moreover, an experiment by using mixed EDTA and HMTA as the complexing agent was performed. The best ZnS thin film with a transmittance of 83% and a denser surface was prepared using HMTA complexing agent.

Influence of Post-Heat Treatment of ZnO:Al Transparent Electrode for Copper Indium Gallium Selenide Thin Film Solar Cell.

Copper indium gallium selenide (CIGS) thin film solar cells have been regarded as a candidate for energy conversion devices owing to their high absorption coefficient, high temperature stability, and low cost. ZnO:Al thin film is commonly used in CIGS solar cells as a window layer. In this study, ZnO:Al films were deposited on glass under various post-heat temperature using RF sputtering to observe the characteristics of ZnO:Al films such as Hall mobility, carrier concentration, and resistivity; subsequently, the ZnO:Al films were applied to a CIGS solar cell as a window. CIGS solar cells fabricated with various ZnO:Al films were analyzed in order to investigate their influence. The test results showed that the improvement of ZnO:Al characteristics affects Jsc and Voc in the solar cell through reduced recombination and increase of optical property.

Thymus vulgaris alleviates UVB irradiation induced skin damage via inhibition of MAPK/AP-1 and activation of Nrf2-ARE antioxidant system.

Solar ultraviolet (UV) radiation-induced reactive oxidative species is mainly responsible for the development of photoageing. Rosmarinic acid was one of the main bioactive components detected in Thymus vulgaris (TV) we extracted. In this study, UVB-induced skin damages have been shown to be ameliorated by treatment with TV in hairless mice (HR-1) skin, demonstrated by decreased matrix metalloproteinases (MMPs) and increased collagen production. However, the underlying molecular mechanism on which TV acted was unclear. We examined the photoprotective effects of TV against UVB and elucidated the molecular mechanism in normal human dermal fibroblasts. Thymus vulgaris remarkably prevented the UVB-induced reactive oxygen species and lactate dehydrogenase. Dose-dependent increase in glutathione, NAD(P)H: quinone oxidoreductase1 and heme oxygenase-1, by TV was confirmed by increased nuclear accumulation of Nrf2. Furthermore, 5-Methoxyindole-2-carboxylic acid was introduced as a specific inhibitor of dihydrolipoyl dehydrogenase (DLD). We demonstrated that Nrf2 expression was regulated by DLD, which was a tricarboxylic acid cycle-associated protein that decreased after UVB exposure. Besides, TV significantly diminished UVB induced phosphorylation of mitogen activated protein kinases pathway, containing extracellular signal-regulated kinase, Jun N-terminal kinase and p38, which consequently reduced phosphorylated c-fos and c-jun. Our results suggest that TV is a potential botanical agent for use against UV radiation-induced oxidative stress mediated skin damages.

Methanol Extract of Bitter Melon Alleviates UVB-Induced MMPs Expression via MAP Kinase and AP-1 Signaling in Human Dermal Fibroblasts in vitro.

Ultraviolet (UV) irradiation leads to photo-damage of the skin, which in turn induces expression of matrix metalloproteinases (MMPs) and reduces type I procollagen. Bitter melon (Momordica charantia L.) has been widely used as a traditional medicine. In this study, we tested the photo-protective effects of methanol extracts of bitter melon pulp (BM) and the mechanism of these effects in normal human dermal fibroblasts (NHDFs). The effects of BM were investigated by measuring the levels of MMP-1, -3 and -9, and type I procollagen following UVB irradiation. We found that BM alleviates UVB-induced MMP-1, -3 and -9 expression at 100 µg/mL (down to 52.0%, 73.5%, and 55.6%, respectively). However, cells treated with 100 µg/mL BM had weakly stimulated type I procollagen expression (up to 130.0%). Moreover, treatment with BM significantly reduced UVB-induced extracellular signal-regulated kinase (ERK), Jun N-terminal kinase (JNK), and p38 phosphorylation, which resulted in decreasing UVB-induced phosphorylation of c-Fos and c-Jun. Therefore, our results suggest that BM is a potential agent for regulating skin photoaging. Copyright © 2016 John Wiley & Sons, Ltd.

The Androgenic Alopecia Protective Effects of Forsythiaside-A and the Molecular Regulation in a Mouse Model.

This study examined the inhibitory effect of forsythiaside-A, a natural substance derived from Forsythia suspensa (F. suspensa), on entry into catagen induced by dihydrotestosterone (DHT) in an androgenic alopecia mouse model. In vitro experiment comparing finasteride with forsythiaside-A showed that forsythiaside-A treatment resulted in a 30% greater inhibition of DHT-induced apoptosis in human hair dermal papilla cell (HHDPCs) and human keratinocytes (HaCaTs). In vivo experiment showed that mouse hair density and thickness were increased by 50% and 30%, respectively, in the forsythiaside-A-treated group when compared to a DHT group. Tissue histological results revealed that the forsythiaside-A-treated group had an increase in size and shape of the hair follicles and a 1.5 times increase in the follicle anagen/telogen ratio when compared to the finasteride group. Western blot examination of TGF-ß2 expression related to apoptosis signaling in mouse skin verified that forsythiaside-A reduced the expression of TGF-ß2 by 75% and suppressed apoptosis by reducing the expression of caspase-9 by 40%, and caspase-3 by 53%, which play an roles up-regulator in the apoptosis signal. The forsythiaside-A group also showed a 60% increase in the Bcl-2/Bax ratio, which is a factor related to mitochondrial apoptosis. Our results indicated that forsythiaside-A prevents apoptosis by similar mechanism with finasteride, but forsythiaside-A is more effective than finasteride. In summary, forsythiaside-A controlled the apoptosis of hair cells and retarded the entry into the catagen phase and therefore represents a natural product with much potential for use as a treatment for androgenic alopecia.

Arthrobacter bambusae sp. nov., isolated from soil of a bamboo grove.

A Gram-stain-positive, aerobic, motile by gliding, rod-shaped bacterial strain, THG-GM18(T), was isolated from soil of a bamboo grove. Strain THG-GM18(T) was able to grow in the presence of up to 6.0 % (w/v) NaCl, at 4-37 °C and at pH 7.0-10.0 in R2A medium. Based on 16S rRNA gene sequence similarity, strain THG-GM18(T) was closely related to species of the genus Arthrobacter. The most closely related strains to strain THG-GM18(T) are Arthrobacter ramosus CCM 1646(T) (98.5 % similarity), Arthrobacter nitroguajacolicus G2-1(T) (98.4 %), Arthrobacter nicotinovorans DSM 420(T) (98.2 %), Arthrobacter aurescens DSM 20116(T) (98.1 %) and Arthrobacter chlorophenolicus A6(T) (98.0 %). Strain THG-GM18(T) possessed chemotaxonomic properties consistent with those of members of the genus Arthrobacter, such as peptidoglycan type A3α (l-Lys-l-Ala-l-Thr-l-Ala), MK-9 as major menaquinone and anteiso- and iso-branched compounds (anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0) as major cellular fatty acids. The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, an unidentified phosphoglycolipid, unidentified phospholipids, unidentified aminolipids, an unidentified glycolipid and unidentified lipids. The G+C content of the genomic DNA was 61.0 mol%. The DNA-DNA relatedness values between strain THG-GM18(T) and its closest phylogenetic neighbours were below 26.0 %. The results of physiological and biochemical tests allowed the differentiation of strain THG-GM18(T) from species of the genus Arthrobacter with validly published names. Arthrobacter bambusae sp. nov. is the proposed name, and the type strain is THG-GM18(T) ( = KACC 17531(T) = JCM 19335(T)).

Pedobacter seoulensis sp. nov., isolated from soil of a bamboo field.

A Gram-stain negative, strictly aerobic, motile by gliding, rod-shaped and yellow pigmented strain THG-G12T was isolated from soil of a bamboo field in Seoul, Republic of Korea. Strain THG-G12T was observed to grow well at 20­28 °C and pH 7.0­7.5 in the absence of NaCl on nutrient agar. Based on 16S rRNA gene sequence comparisons, strain THG-G12T was found to be most closely related to Pedobacter ginsengisoli Gsoil 104T (97.5 % sequence similarity), Pedobacter steynii WB2.3-45T (97.4 %), Pedobacter metabolipauper WB2.3-71T (97.2 %), Pedobacter nyackensis NWG-II14T (97.2 %), Pedobacter caeni LMG 22862T (97.1 %) and Pedobacter duraquae WB2.1-25T (97.0 %), but DNA relatedness between strain THG-G12T and its phylogenetically closest neighbours was below 9.5 %. The G+C content of the genomic DNA was determined to be 39.9 mol%. The only isoprenoid quinone detected in strain THG-G12T was menaquinone-7 (MK-7). The major component in the polyamine pattern was sym-homospermidine. The major polar lipids were found to be phosphatidylethanolamine, unidentified phosphoglycolipids, unidentified aminophosphoglycolipids, unidentified aminolipids and unidentified lipids. Strain THG-G12T showed the presence of two ceramide phosphorylethanolamines (CerPE-2' and CerPE-2″), dihydrosphingosines and an unidentified ceramide as the major ceramide. The major fatty acids were identified as summed feature 3 (as defined by the MIDI system; C16:1 ω7c and/or C16:1 ω6c) and iso-C15:0. These data support the affiliation of strain THG-G12T to the genus Pedobacter. The results of physiological and biochemical tests enabled strain THG-G12T to be differentiated genotypically and phenotypically from the recognized species of the genus Pedobacter. Therefore, the novel isolate represents a novel species, for which the name Pedobacter seoulensis sp. nov. is proposed, with THG-G12T as the type strain (=KACC 17529T =JCM 19363T).

Sphingomonas kyeonggiense sp. nov., isolated from soil of a ginseng field.

A Gram-staining negative bacterium, THG-DT81(T), which was isolated from soil of a ginseng field, was investigated using a polyphasic taxonomic approach. Cells were oxidase- and catalase-positive, aerobic, rod-shaped and motile with one polar flagellum. Strain THG-DT81(T) grew optimally at pH 7.0 and in the absence of NaCl on trypticase soy agar. Its optimum growth temperature was 25-28 °C. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain THG-DT81(T) belongs to the family Sphingomonadaceae and was related to Sphingomonas pituitosa EDIV(T) (98.0 % similarity), S. leidyi ATCC 15260(T) (97.8 %), S. trueperi LMG 2142(T) (97.1 %), S. azotifigens NBRC 15497(T) (97.1 %), S. koreensis JSS26 (T) (97.1 %) and S. dokdonensis DS-4(T) (97.0 %). Strain THG-DT81(T) contained Q-10 as the predominant ubiquinone and C18:1 ω7c and C16:0 as the major fatty acids. The G+C content of the genomic DNA was determined to be 66.8 mol %. The major component in the polyamine pattern was identified as sym-homospermidine. The major polar lipids detected in strain THG-DT81(T) were sphingoglycolipid, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and phosphatidylcholine. The DNA-DNA relatedness values of the strain THG-DT81(T) and its closest phylogenetically neighbors were below 21 %. The phenotypic characteristics and genotypic data demonstrated the affiliation of strain THG-DT81(T) to the genus Sphingomonas. On the basis of the polyphasic taxonomic data presented, strain THG-DT81(T) is described as a novel species of genus Sphingomonas, for which the name Sphingomonas kyeonggiense sp. nov. is proposed. The type strain is THG-DT81(T) (= KACC 17173(T) = JCM 18825(T)).

Ginsenoside F2 reduces hair loss by controlling apoptosis through the sterol regulatory element-binding protein cleavage activating protein and transforming growth factor-ß pathways in a dihydrotestosterone-induced mouse model.

This study was conducted to test whether ginsenoside F2 can reduce hair loss by influencing sterol regulatory element-binding protein (SREBP) cleavage-activating protein (SCAP) and the transforming growth factor beta (TGF-ß) pathway of apoptosis in dihydrotestosterone (DHT)-treated hair cells and in a DHT-induced hair loss model in mice. Results for ginsenoside F2 were compared with finasteride. DHT inhibits proliferation of hair cells and induces androgenetic alopecia and was shown to activate an apoptosis signal pathway both in vitro and in vivo. The cell-based 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that the proliferation rates of DHT-treated human hair dermal papilla cells (HHDPCs) and HaCaTs increased by 48% in the ginsenoside F2-treated group and by 12% in the finasteride-treated group. Western blot analysis showed that ginsenoside F2 decreased expression of TGF-ß2 related factors involved in hair loss. The present study suggested a hair loss related pathway by changing SCAP related apoptosis pathway, which has been known to control cholesterol metabolism. SCAP, sterol regulatory element-binding protein (SREBP) and caspase-12 expression in the ginsenoside F2-treated group were decreased compared to the DHT and finasteride-treated group. C57BL/6 mice were also prepared by injection with DHT and then treated with ginsenoside F2 or finasteride. Hair growth rate, density, thickness measurements and tissue histotological analysis in these groups suggested that ginsenoside F2 suppressed hair cell apoptosis and premature entry to catagen more effectively than finasteride. Our results indicated that ginsenoside F2 decreased the expression of TGF-ß2 and SCAP proteins, which have been suggested to be involved in apoptosis and entry into catagen. This study provides evidence those factors in the SCAP pathway could be targets for hair loss prevention drugs.

Gallic acid regulates skin photoaging in UVB-exposed fibroblast and hairless mice.

Ultraviolet (UV) radiation is the primary factor in skin photoaging, which is characterized by wrinkle formation, dryness, and thickening. The mechanisms underlying skin photoaging are closely associated with degradation of collagen via upregulation of matrix metalloproteinase (MMP) activity, which is induced by reactive oxygen species (ROS) production. Gallic acid (GA), a phenolic compound, possesses a variety of biological activities including antioxidant and antiinflammatory activities. We investigated the protective effects of GA against photoaging caused by UVB irradiation using normal human dermal fibroblasts (NHDFs) in vitro and hairless mice in vivo. The production levels of ROS, interlukin-6, and MMP-1 were significantly suppressed, and type I procollagen expression was stimulated in UVB-irradiated and GA-treated NHDFs. GA treatment inhibited the activity of transcription factor activation protein 1. The effects of GA following topical application and dietary administration were examined by measuring wrinkle formation, histological modification, protein expression, and physiological changes such as stratum corneum hydration, transepidermal water loss, and erythema index. We found that GA decreased dryness, skin thickness, and wrinkle formation via negative modulation of MMP-1 secretion and positive regulation of elastin, type I procollagen, and transforming growth factor-ß1. Our data indicate that GA is a potential candidate for the prevention of UVB-induced premature skin aging.

Lactobacillus yonginensis sp. nov., a lactic acid bacterium with ginsenoside converting activity isolated from Kimchi.

A Gram-reaction-positive, non-motile, non-spore-forming, catalase-negative, facultatively anaerobic, rod-shaped, ß-glucosidase-producing lactic acid bacterium, designated strain THK-V8(T), was isolated from the Korean fermented food, Kimchi, and its taxonomic position was investigated by using a polyphasic approach. Strain THK-V8(T) was able to grow at 4-40 °C (optimum, 30 °C) and pH 4.0-7.0 (optimum, pH 6.0). Strain THK-V8(T) had the ability to transform ginsenoside Rb1 to Rd. On the basis of 16S rRNA gene sequence similarity data, strain THK-V8(T) was shown to belong to the genus Lactobacillus. Strain THK-V8(T) was related to Lactobacillus koreensis DCY50(T) (98.8% sequence similarity), Lactobacillus parabrevis LMG 11984(T) (97.7%), Lactobacillus senmaizukei L13(T) (97.5%), Lactobacillus hammesii TMW1.1236(T) (97.3%) and Lactobacillus brevis ATCC 14687(T) (97.2%). Subsequently, sequence analysis of the RNA polymerase alpha subunit gene (rpoA) confirmed that strain THK-V8(T) showed a maximum rpoA gene sequence similarity value of 93% with Lactobacillus brevis LMG 6906(T). The G+C content of the genomic DNA was 47.8 mol%. The DNA-DNA hybridization values between strain THK-V8(T) and Lactobacillus parabrevis DCY50(T) and Lactobacillus parabrevis LMG 11984(T) were 46.1 ± 4.9% and 10.6 ± 2.9%, respectively. The major fatty acids were summed feature 7 (comprised of C(19:0) cyclo ω10c/19ω6), C(14:0), C(16:0) and C(18:1)ω9c. The cell wall peptidoglycan was of the A4α L-Lys-D-Asp type. The phenotypic and molecular properties indicated that strain THK-V8(T) represents a novel species within the genus Lactobacillus, for which the name Lactobacillus yonginensis sp. nov. is proposed. The type strain is THK-V8(T) ( =KACC 16236(T) =JCM 18023(T)).

Pedobacter ginsenosidimutans sp. nov., with ginsenoside-converting activity.

A Gram-reaction-negative, strictly aerobic, non-motile, non-spore-forming and rod-shaped bacterial strain, designated THG-45(T), was isolated from soil of a ginseng field of Pocheon province in the Republic of Korea and its taxonomic position was investigated by a polyphasic approach. Growth occurred at 4-30 °C, at pH 5.5-9.0 and with 0-2 % (w/v) NaCl on nutrient agar. On the basis of 16S rRNA gene sequence similarity, strain THG-45(T) was shown to belong to the genus Pedobacter and was related to Pedobacter borealis G-1(T) (98.8 %), P. alluvionis NWER-II11(T) (97.9 %), P. agri PB92(T) (97.9 %), P. terrae DS-57(T) (97.5 %), P. suwonensis 15-52(T) (97.4 %), P. sandarakinus DS-27(T) (97.0 %) and P. soli 15-51(T) (97.0 %), but DNA relatedness between strain THG-45(T) and these strains was below 36 %. The G+C content of the genomic DNA was 39 mol%. The only isoprenoid quinone detected in strain THG-45(T) was menaquinone-7 (MK-7). The predominant fatty acids were iso-C15 : 0, summed feature 3 (comprising C16 : 1ω6c and/or C16 : 1ω7c) and iso-C17 : 0 3-OH, and the major polar lipids were phosphatidylethanolamine and an unidentified aminophosphoglycolipid. Phenotypic data and phylogenetic inference supported the affiliation of strain THG-45(T) to the genus Pedobacter, and a number of biochemical tests differentiated strain THG-45(T) from the recognized species of the genus Pedobacter. Therefore, the novel isolate represents a novel species, for which the name Pedobacter ginsenosidimutans sp. nov. is proposed, with THG-45(T) as the type strain ( = KACC 14530(T) = JCM 16721(T)).

Flavobacterium kyungheensis sp. nov., isolated from soil of a ginseng field.

A Gram-staining negative, strictly aerobic, motile by gliding, non-spore-forming, pale yellow pigmented and rod-shaped bacterium designated strain THG-107(T) was isolated from soil of a ginseng field on Ganghwa Island in the Republic of Korea and its taxonomic position was investigated by using a polyphasic study. Growth of strain THG-107(T) was found to occur at 4-37 °C (optimum, 20-30 °C), at pH 5.5-10 (optimum, pH 7.0) and in the presence of 0-1 % (w/v) NaCl (optimum, absence) on R2A agar. On the basis of 16S rRNA gene sequence similarity, strain THG-107(T) was shown to belong to the family Flavobacteriaceae and was related to Flavobacterium denitrificans ED5(T) (99.1 % similarity). The G+C content of the genomic DNA was determined to be 34.2 mol%. These results are consistent with characteristics of members of the genus Flavobacterium. The only isoprenoid quinone detected in strain THG-107(T) was menaquinone-6 (MK-6) and the major polyamine was identified as homospermidine (82.9 %). The major polar lipid detected was phosphatidylethanolamine and the major cellular fatty acids were identified as iso-C15:0 (26.3 %), iso-C17:0 3OH (12.6 %) and summed feature 3 (comprising C16:1 ω7c and/or C16:1 ω6c; 11.6 %). Flexirubin-type pigments were found to be present. Strain THG-107(T) has ß-glucosidase activity to convert ginsenosides Rb1 and Rd into Gyp17 and F2. DNA-DNA hybridization with F. denitrificans ED5(T) was 52 %. Strain THG-107(T) could be distinguished from F. denitrificans ED5(T) and the other species of the genus Flavobacterium by its phylogenetic and genetic distinctiveness and by several phenotypic properties. Therefore, strain THG-107(T) is considered to represent a novel species in the genus Flavobacterium, for which the name Flavobacterium kyungheensis sp. nov. is proposed (type strain THG-107(T) = KACC 16219(T) = LMG 26575(T)).

Hair growth activity of Crataegus pinnatifida on C57BL/6 mouse model.

Crataegus pinnatifida has a long history of use in traditional oriental herbal medicine to stimulating digestion and improving blood circulation. Based on nutrition of hair, the present study was conducted to assess the effect of C. pinnatifida extract on hair growth using mouse model and its mechanisms of action. The C. pinnatifida extract containing the contents of total polyphenol of 5.88□0.82 g gallic acid/100 g extract and proanthocyanidin of 9.15□1.58 mg cyaniding chloride/100 g extract was orally administered daily at a dosage of 50 mg/kg weight to the 7-week-old C57BL/6 mice in telogen. The C. pinnatifida extract promoted hair growth by inducing anagen phase in mice in telogen, reflected by color of skin, thickness of hair shaft, and density of hair. The ratio of anagento telogen was determined by shape of hair follicles in vertically sectioned slide and increased by oral administration of C. pinnatifida extract. The number and the size of hair follicles were also enlarged, indicating anagen phase induction. The proliferation of human dermal papilla cells (hDPC) was accelerated by addition of C. pinnatifida extract, which activated the signaling of mitogen-activated protein kinases (Erk, p-38, and JNK) and Akt. Moreover, the ratio of Bcl-2/Bax as the determinant of cell fate was also raised in skin. These results suggest that the C. pinnatifida extract promotes hair growth by inducing anagen phase, which might be mediated by the activation of cellular signalings that enhance the survival of cultured hDPC and the increase of the ratio of Bcl-2 to Bax that protects cells against cell death.