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1.
Photochem Photobiol Sci ; 23(5): 987-996, 2024 May.
Article in English | MEDLINE | ID: mdl-38662174

ABSTRACT

Pycnoporus sanguineus is a fungus of the phylum Basidiomycota that has many applications in traditional medicine, modern pharmaceuticals, and agricultural industries. Light plays an essential role in the metabolism, growth, and development of fungi. This study evaluated the mycelial growth and antioxidant and anti-inflammatory activities in P. sanguineus fermentation broth (PFB) cultured under different wavelengths of LED irradiation or in the dark. Compared to the dark cultures, the dry weight of mycelia in red- and yellow-light cultures decreased by 37 and 35% and the yields of pigments increased by 30.92 ± 2.18 mg and 31.75 ± 3.06 mg, respectively. Compared with the dark culture, the DPPH free radical scavenging ability, ABTS+ free radical scavenging capacity, and reducing power of yellow-light cultures increased significantly, and their total phenolic content peaked at 180.0 ± 8.34 µg/mL. However, the reducing power in blue-light cultures was significantly reduced, though the total phenol content did not vary with that of dark cultures. In LPS- and IFN-γ-stimulated RAW 264.7 cells, nitrite release was significantly reduced in the red and yellow light-irradiated PFB compared with the dark culture. In the dark, yellow-, and green-light cultures, TNF-α production in the inflamed RAW 264.7 cells was inhibited by 62, 46, and 14%, respectively. With red-, blue-, and white-light irradiation, TNF-α production was significantly enhanced. Based on these results, we propose that by adjusting the wavelength of the light source during culture, one can effectively modulate the growth, development, and metabolism of P. sanguineus.


Subject(s)
Antioxidants , Light , Pycnoporus , Mice , Animals , Antioxidants/pharmacology , Antioxidants/chemistry , Antioxidants/metabolism , RAW 264.7 Cells , Pycnoporus/metabolism , Immunologic Factors/pharmacology , Immunologic Factors/chemistry , Lipopolysaccharides/pharmacology , Lipopolysaccharides/antagonists & inhibitors , Picrates/antagonists & inhibitors , Picrates/chemistry , Immunomodulating Agents/pharmacology , Immunomodulating Agents/chemistry , Biphenyl Compounds/antagonists & inhibitors , Biphenyl Compounds/chemistry , Biphenyl Compounds/pharmacology
2.
Toxicol In Vitro ; 86: 105511, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36336209

ABSTRACT

Chronic exposure of skin to ultraviolet (UV) radiation is responsible for skin ageing, which includes degradation of the epidermal and dermal layers. Filtering UV light is key in the sunscreen industry. We studied the effects of organic UV filters on hyaluronan (HA) metabolism and skin hydration in human HaCaT keratinocytes. The gene expression of HA receptors, HA synthase (HAS), hyaluronidase (HYAL), and water channel aquaporin 3 (AQP3) was evaluated by quantitative RT-PCR. The state of oxidative stress was determined by measuring the intracellular levels of reactive oxygen species (ROS). The results showed that five organic UV filters reduced the extracellular contents of HA, and a phosphatidylinositol 3-kinase (PI3K) inhibitor partially restored the decreased HA levels after octinoxate, octocrylene, and oxybenzone treatment. The expression levels of HA receptors, including cluster of differentiation 44 (CD44), receptor for hyaluronic acid-mediated motility (RHAMM), and toll-like receptors (TLRs), were determined. Avobenzone, octinoxate, oxybenzone, and padimate O exerted inhibitory effects on RHAMM expression. Oxybenzone led to a significant increase in CD44 and AQP3 expression. Both octinoxate and octocrylene increased TLR4 expression but decreased ROS accumulation by activating the PI3K pathway. However, the organic UV filters differentially regulated the mRNA expression of HAS and HYAL. Taken together, these results suggest that certain organic UV filters regulate HA metabolism in human keratinocytes in a PI3K pathway-dependent manner.


Subject(s)
Hyaluronic Acid , Phosphatidylinositol 3-Kinase , Humans , Phosphatidylinositol 3-Kinase/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Reactive Oxygen Species/metabolism , Keratinocytes , Ultraviolet Rays , Hyaluronoglucosaminidase/metabolism
3.
Int J Biol Macromol ; 209(Pt A): 506-512, 2022 Jun 01.
Article in English | MEDLINE | ID: mdl-35398387

ABSTRACT

Gum Arabic, a mixture of polysaccharide and glycoprotein, is used as an emulsifying stabilizer in the food industry. It might have immunomodulatory effects. We hypothesized that the combination of IFN-γ and Gum Arabic promotes the production of pro-inflammatory factors in RAW 264.7 cells. Treatment of RAW 264.7 cells with the combination of 3% Gum Arabic and 40 ng/mL IFN-γ resulted in a drastic increase (320%) in nitric oxide production compared with that induced by IFN-γ alone. PGE-2 was produced after the cells were treated with 3% Gum Arabic and 40 ng/mL IFN-γ for 6 h. Gum Arabic and IFN-γ increased the production of iNOS and COX-2 proteins, and triggered TNF-α release. Apart from TNF-α, the release of both G-CSF and IL-6 increased by more than 100 times. The release of IL-3, RANTES, and IL-10 increased by more than ten times. Gum Arabic and IFN-γ also increased the secretion of IL-10, IL-1α, IL-1ß, IL-13, KC, IL-5, IL-4, IL-12, Eotaxin, IL-9, MCP-1, and ROS. Cytokines associated with M1 polarization of macrophages such as TNF-α, IL-1ß, IL-12, NO, and ROS were induced by Gum Arabic and IFN-γ. Our findings help to explore the inflammatory reaction caused by Gum Arabic in cosmetics.


Subject(s)
Interleukin-10 , Tumor Necrosis Factor-alpha , Cytokines/metabolism , Gum Arabic/pharmacology , Interferon-gamma/metabolism , Interferon-gamma/pharmacology , Interleukin-10/metabolism , Interleukin-12/metabolism , Macrophages , Nitric Oxide/metabolism , Reactive Oxygen Species/metabolism , Tumor Necrosis Factor-alpha/metabolism
4.
Phytother Res ; 33(4): 1104-1113, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30734961

ABSTRACT

Myrica rubra Sieb. et Zucc. (Myricaceae), known as Chinese bayberry, is traditionally used as folk medicine in Asian countries. The interaction of Propionibacterium acnes signalling with sebocytes is considered important in the pathogenesis of acne. In the present study, extracts and active compounds of Chinese bayberry were used to determine chemical antioxidant activity and anti-inflammatory effects in P. acnes-stimulated human SZ95 sebocytes. A high-performance liquid chromatography with electrochemical detection system was used to analyse the phenolic composition of bayberry extracts. Accordingly, the flavonols, myricitrin and myricetin, were found to be abundant in the unhydrolysed and hydrolysed extracts of Chinese bayberry fruits, respectively. The anthocyanin cyanidin-3-glucoside was also predominantly found in the unhydrolysed extracts. Quantification of human inflammatory cytokines indicated that cell-free extracts of P. acnes stimulated IL-8 and IL-6 production, which was inhibited by myricetin, rather than its glycoside or anthocyanin. Myricetin also exhibited inhibitory effects in P. acnes-stimulated gene expression of Toll-like receptor (TLR) 2 and protein phosphorylation of p70 S6 kinase. In conclusion, myricetin shows a suppressive effect on P. acnes-induced cytokine production through regulation of the TLR and mammalian target of rapamycin pathways. Myricetin goes beyond previous research findings to potentially modulate inflammatory signalling in human sebocytes. These results will be valuable in developing anti-inflammatory agents against skin acne.


Subject(s)
Cytokines/drug effects , Flavonoids/therapeutic use , Myrica/chemistry , Plant Extracts/chemistry , Propionibacterium acnes/drug effects , Drugs, Chinese Herbal , Flavonoids/pharmacology , Humans , Plant Extracts/pharmacology
5.
Int J Biol Macromol ; 114: 181-186, 2018 Jul 15.
Article in English | MEDLINE | ID: mdl-29572144

ABSTRACT

Locust bean gum (LBG) galactomannan has been claimed to have applications in the biopharmaceutical field. However, the effects of LBG galactomannan on immunomodulatory aspects are not yet clear. The purpose of this study was to over-express thermostable ß-d-mannanase from the thermophilic actinomycete Thermobifida fusca BCRC 19214 using a Pichia pastoris expression system. The maximum intracellular ß-d-mannanase activity obtained from the cell-free extract was approximately 40.0U/mL after 72h of cultivating a P. pastoris transformant (pPICZ-man) induced with methanol. Hydrolysis of native LBG galactomannan with 8U/mL ß-d-mannanase for 24h significantly decreased the weight-average molecular weight of LBG galactomannan from 5,580,010 to 3188. Native and hydrolyzed LBG galactomannan in a range of 0-0.2% did not trigger significant cytotoxicity after 24h of treatment compared with the control. The native LBG galactomannan stimulated RAW 264.7 cells to produce cytokine TNF-α dose-dependently, but there was no significant IL-1ß or nitric oxide production. The native LBG galactomannan also stimulated ß-hexosaminidase secretion in RBL-2H3 cells. After the native LBG galactomannan was hydrolyzed with ß-d-mannanase, all of the immunological properties disappeared. These results suggest the possible immunomodulatory effects of native LBG galactomannan.


Subject(s)
Actinomyces/enzymology , Fungal Proteins/chemistry , Galactans/chemistry , Interleukin-1beta/metabolism , Mannans/chemistry , Nitric Oxide/metabolism , Plant Gums/chemistry , Tumor Necrosis Factor-alpha/metabolism , beta-Mannosidase/chemistry , Actinomyces/genetics , Animals , Fungal Proteins/genetics , Galactose/analogs & derivatives , Hydrolysis , Mannans/pharmacology , Mice , RAW 264.7 Cells , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , beta-Mannosidase/genetics
6.
J Food Drug Anal ; 25(4): 881-889, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28987365

ABSTRACT

This study analyzed 26 commercially available essential oils and their major chemical components to determine their antioxidant activity levels by measuring their total phenolic content (TPC), reducing power (RP), ß-carotene bleaching (BCB) activity, trolox equivalent antioxidant capacity (TEAC), and 1,1-diphenyl-2-picrylhydrazyl free radical scavenging (DFRS) ability. The clove bud and thyme borneol essential oils had the highest RP, BCB activity levels, and TPC values among the 26 commercial essential oils. Furthermore, of the 26 essential oils, the clove bud and ylang ylang complete essential oils had the highest TEAC values, and the clove bud and jasmine absolute essential oils had the highest DFRS ability. At a concentration of 2.5 mg/mL, the clove bud and thyme borneol essential oils had RP and BCB activity levels of 94.56% ± 0.06% and 24.64% ± 0.03% and 94.58% ± 0.01% and 89.33% ± 0.09%, respectively. At a concentration of 1 mg/mL, the clove bud and thyme borneol essential oils showed TPC values of 220.00 ± 0.01 and 69.05 ± 0.01 mg/g relative to gallic acid equivalents, respectively, and the clove bud and ylang ylang complete essential oils had TEAC values of 809.00 ± 0.01 and 432.33 ± 0.01 µM, respectively. The clove bud and jasmine absolute essential oils showed DFRS abilities of 94.13% ± 0.01% and 78.62% ± 0.01%, respectively. Phenolic compounds of the clove bud, thyme borneol and jasmine absolute essential oils were eugenol (76.08%), thymol (14.36%) and carvacrol (12.33%), and eugenol (0.87%), respectively. The phenolic compounds in essential oils were positively correlated with the RP, BCB activity, TPC, TEAC, and DFRS ability.


Subject(s)
Antioxidants/chemistry , Oils, Volatile/chemistry , Plant Extracts/chemistry , Antioxidants/economics , Chromatography, Gas , Oils, Volatile/economics , Plant Extracts/economics
7.
Int J Biol Macromol ; 82: 1-6, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26476245

ABSTRACT

Native konjac glucomannan was used as the substrate for thermophilic actinomycetes, Thermobifida fusca BCRC19214, to produce ß-mannanase. The ß-mannanase was purified and five internal amino acid sequences were determined by LC-MS/MS. These sequences had high homology with the ß-mannanase from T. fusca YX. The tfm gene which encoded the ß-mannanase was cloned, sequenced and heterologous expressed in Yarrowia lipolytica P01 g expression system. Recombinant heterologous expression resulted in extracellular ß-mannanase production at levels as high as 3.16 U/ml in the culture broth within 48 h cultivation. The recombinant ß-mannanase from Y. lipolytica transformant had superior thermal property. The optimal temperature of the recombinant ß-mannanase from Y. lipolytica transformant (pYLSC1-tfm) was 80°C. When native konjac glucomannan was incubated with the recombinant ß-mannanase from Y. lipolytica transformant (pYLSC1-tfm) at 50°C, there was a fast decrease of viscosity happen during the initial phase of reaction. This viscosity reduction was accompanied by an increase of reducing sugars. The surface of konjac glucomannan film became smooth. After 24h of treatment, the DPw of native konjac glucomannan decreased from 6,435,139 to 3089.


Subject(s)
Mannans/chemistry , Yeasts/enzymology , beta-Mannosidase/chemistry , Enzyme Activation , Enzyme Stability , Fermentation , Hydrolysis , Recombinant Proteins/chemistry , Temperature , Yeasts/genetics , beta-Mannosidase/biosynthesis , beta-Mannosidase/isolation & purification
8.
Life Sci ; 139: 123-31, 2015 Oct 15.
Article in English | MEDLINE | ID: mdl-26341693

ABSTRACT

AIMS: Propionibacterium acnes has been considered to influence the acne lesions. The present study intended to elucidate the underlying signaling pathways of P. acnes in human sebaceous gland cells relative to the generation of proinflammatory cytokines. MAIN METHODS: Cell-free extracts of P. acnes under stationary growth phase were co-incubated with human immortalized SZ95 sebocytes. Then, cell-free P. acnes extracts-induced cytokine expression was evaluated by measuring mRNA and protein levels using quantitative RT-PCR and ELISA. Changes of phosphorylated cell signaling proteins and transcription factors were measured by Western blots and Milliplex assay. The interactive molecular mechanisms of P. acnes and sebocytes were examined through use of shRNA and the specific inhibitors of signaling pathways. KEY FINDINGS: Cell-free extracts of P. acnes significantly stimulated secretion of interleukin (IL)-8 and IL-6 in SZ95 sebocytes. The degradation of IκB-α and increased phosphorylation of IκB-α, p38 mitogen activated protein kinase (MAPK), CREB, and STAT3 were demonstrated. Quantitative RT-PCR measurements revealed that gene expression of IL-8 and Toll-like receptor 2 (TLR2) was enhanced by cell-free extracts of P. acnes. In addition, the NF-κB inhibitor BMS345541, p38 MAPK inhibitor SB203580, or anti-TLR2 neutralizing antibody prevented cell-free P. acnes extracts-induced secretion of IL-8. Knockdown of TLR2 using shRNA exerted similar inhibitory effects on IL-8 expression. Moreover, inhibition of STAT3 activity by STA-21 enhanced P. acnes-mediated secretion of IL-8. SIGNIFICANCE: Cell-free extracts of P. acnes are capable to activate NF-κB and p38 MAPK pathways and up-regulate secretion of IL-8 through TLR2-dependent signaling in human SZ95 sebocytes.


Subject(s)
Interleukin-8/immunology , NF-kappa B/immunology , Propionibacterium acnes/immunology , Sebaceous Glands/microbiology , Toll-Like Receptor 2/immunology , p38 Mitogen-Activated Protein Kinases/immunology , Acne Vulgaris/immunology , Acne Vulgaris/microbiology , Cell Line , Cytokines/immunology , Humans , Sebaceous Glands/cytology , Sebaceous Glands/immunology , Signal Transduction
9.
Curr Pharm Biotechnol ; 16(12): 1053-62, 2015.
Article in English | MEDLINE | ID: mdl-26306747

ABSTRACT

Elastic fibers are major constituents of the extracellular matrix (ECM) in dynamic tissues in the human body, and regulation of elastin and fibrillin-1 expression mediates the formation of these fibers. Traditional assays for the measurement of elastin and fibrillin-1, such as western blotting, Luna staining and immunostaining, are relatively complex and time-consuming. Thus, a relatively simple assay system that also provides rational results is urgently needed. In the study, we aimed to develop a human cell-based assay system that can be used to analyze functional compounds using the promoters of elastin (ELN) and fibrillin-1 (FBN1) genes integrated with a secreted alkaline phosphatase (SEAP) reporter in normal human fibroblast cells. We used this system to assess anti-aging compounds. We used several regulators of elastinogenesis, including retinol, coenzyme Q10, deoxyArbutin and Elestan(TM) (Manilkara multinervis leaf extract), to verify the efficacy of this assay system. Our results demonstrate that this assay system can be used as a fast and realistic method for identifying anti-aging components for future use in foods, cosmetics and drugs.


Subject(s)
Aging , Alkaline Phosphatase/metabolism , Elastin/metabolism , Fibroblasts/metabolism , Microfilament Proteins/metabolism , Cell Line , Elastin/genetics , Extracellular Matrix/metabolism , Fibrillin-1 , Fibrillins , Humans , Microfilament Proteins/genetics
10.
Enzyme Microb Technol ; 68: 1-9, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25435499

ABSTRACT

There are two established depigmenting agent assays currently in use. However, these methods are unreliable and time-consuming. Therefore, it will be valuable to establish a better assay system for depigmenting agent analysis. In this study, we established a melanogenesis regulation assay system using a fluorescent protein reporter combined with the promoters for the microphthalmia-associated transcription factor (MITF), tyrosinase (Tyr) and dopachrome tautomerase (Dct) genes in MeWo human melanoma cells. We used several melanogenesis regulators, including theophylline, hesperetin, arbutin and rottlerin, to confirm the function of this assay system. The established MeWo/pMITF-EGFP, MeWo/pTyr-EGFP and MeWo/pDct-EGFP stable cells integrated the pMITF-EGFP, pTyr-EGFP and pDct-EGFP plasmids into their genomic DNA. These stably transfected cells were used to examine alterations in the expression of the MITF, Tyr and Dct genes. All of the tested compounds, including theophylline, hesperetin, arbutin and rottlerin, could be analyzed in the stable cells, producing reliable results. Therefore, we believe that this melanogenesis regulation assay system can be used as a rapid and reliable assay system to analyze the regulation of melanogenesis by many known or unknown compounds.


Subject(s)
Green Fluorescent Proteins/analysis , Melanins/biosynthesis , Melanoma/pathology , Promoter Regions, Genetic/genetics , Skin Lightening Preparations/pharmacology , Acetophenones/pharmacology , Arbutin/pharmacology , Benzopyrans/pharmacology , Cell Line, Tumor , Drug Evaluation, Preclinical/methods , Enzyme Induction/drug effects , Gene Expression Regulation, Neoplastic/drug effects , Genes, Reporter , Genes, Synthetic , Green Fluorescent Proteins/genetics , Hesperidin/pharmacology , Humans , Intramolecular Oxidoreductases/biosynthesis , Intramolecular Oxidoreductases/genetics , Melanins/analysis , Melanoma/genetics , Melanoma, Experimental/pathology , Microphthalmia-Associated Transcription Factor/biosynthesis , Microphthalmia-Associated Transcription Factor/genetics , Microscopy, Fluorescence , Monophenol Monooxygenase/biosynthesis , Monophenol Monooxygenase/genetics , Neoplasm Proteins/biosynthesis , Neoplasm Proteins/genetics , RNA, Messenger/biosynthesis , Recombinant Fusion Proteins/analysis , Theophylline/pharmacology , Transfection
11.
AMB Express ; 3(1): 49, 2013 Aug 28.
Article in English | MEDLINE | ID: mdl-23985268

ABSTRACT

Laccases are diphenol oxidases that have numerous applications to biotechnological processes. In this study, the laccase was produced from the thermophilic actinomycetes, Thermobifida fusca BCRC 19214. After 36 h of fermentation in a 5-liter fermentor, the culture broth accumulated 4.96 U/ml laccase activity. The laccase was purified 4.64-fold as measured by specific activity from crude culture filtrate by ultrafiltration concentration, Q-Sepharose FF and Sephacryl™ S-200 column chromatography. The overall yield of the purified enzyme was 7.49%. The molecular mass of purified enzyme as estimated by SDS-PAGE and by gel filtration on Sephacryl™ S-200 was found to be 73.3 kDa and 24.7 kDa, respectively, indicating that the laccase from T. fusca BCRC 19214 is a trimer. The internal amino acid sequences of the purified laccase, as determined by LC-MS/MS, had high homology with a superoxide dismutase from T. fusca YX. Approximately 95% of the original activity remained after treatment at 50°C for 3 h. and approximately 75% of the original activity remained after treatment at pH 10.0 for 24 h. This laccase could oxidize dye intermediates, especially 2,6-dimethylphenylalanine and p-aminophenol, to produce coloring. This is the first report on laccase properties from thermophilic actinomycetes. These properties suggest that this newly isolated laccase has potential for specific industrial applications.

12.
Chem Biol Interact ; 205(3): 157-64, 2013 Oct 05.
Article in English | MEDLINE | ID: mdl-23872139

ABSTRACT

Fenofibrate and ciglitazone belong to the classes of fibrates and thiazolidinediones, respectively. Their pharmacological actions on peroxisome proliferator-activated receptors (PPARs) present a potential therapy for hyperlipidemia and hyperglycemia. However, the melanogenesis affected by PPAR ligands in melanocytes has not been well investigated. By determining the melanin content of cells treated with PPAR agonists, we showed that fenofibrate significantly reduced melanin synthesis, but its major active metabolite, fenofibric acid, did not. Notably, the suppression of melanogenesis by fenofibrate could not be prevented by the PPARα specific antagonist GW6471. In addition, T0901317, a liver X receptor (LXR) agonist, restored the antimelanogenic activity of fenofibrate. Accordingly, fenofibrate may suppress melanogenesis through a PPARα-independent pathway. Treatment of cells with fenofibrate led to the down-regulated gene expression of melanocortin 1 receptor (MC1R). Fenofibrate also attenuated the dihydroxyphenylalanine (DOPA)-staining activity and expression of tyrosinase as well as the expression of microphthalmia-associated transcription factor (MITF). The phosphorylation of p38 mitogen-activated protein kinase (MAPK) was stimulated by fenofibrate. Furthermore, the p38 MAPK inhibitor SB203580 prevented the repressive effects of fenofibrate on the melanin production. Taken together, the results of the present study suggest that fenofibrate inhibits melanin synthesis via the down-regulation of MC1R, the up-regulation of p38 MAPK, and interference with LXR signaling pathways to decrease the expression of tyrosinase in B16-F10 melanoma cells.


Subject(s)
Fenofibrate/pharmacology , Melanins/biosynthesis , Melanoma, Experimental/drug therapy , Melanoma, Experimental/metabolism , Blotting, Western , Cell Line, Tumor , Cell Survival/drug effects , Humans , Imidazoles/pharmacology , MAP Kinase Signaling System , Melanoma, Experimental/enzymology , Microphthalmia-Associated Transcription Factor/genetics , Microphthalmia-Associated Transcription Factor/metabolism , Oxazoles/pharmacology , PPAR alpha/antagonists & inhibitors , PPAR alpha/metabolism , Pyridines/pharmacology , RNA/chemistry , RNA/genetics , Real-Time Polymerase Chain Reaction , Tyrosine/analogs & derivatives , Tyrosine/pharmacology
13.
Biosci Biotechnol Biochem ; 77(5): 1127-30, 2013.
Article in English | MEDLINE | ID: mdl-23649246

ABSTRACT

Arbutin and deoxy arbutin may release hydroquinone under some conditions. We therefore investigated the photostability of arbutin and deoxy arbutin in an aqueous solution. The results revealed arbutin and deoxy arbutin to be photolabile in an aqueous solution. Deoxy arbutin was less stable than arbutin when exposed to UV radiation. The hydroquinone concentration was also increased during the radiation period in both solutions. Benzophenone-4 could clearly improve the photostability of arbutin during the period of UV radiation, but only slightly enhance the photostability of deoxy arbutin.


Subject(s)
Arbutin/analogs & derivatives , Benzophenones/chemistry , Photochemical Processes , Sunscreening Agents/chemistry , Ultraviolet Rays , Water/chemistry , Arbutin/chemistry , Drug Stability , Solubility
14.
Appl Microbiol Biotechnol ; 97(20): 8977-86, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23377789

ABSTRACT

Thermobifida fusca is a moderately thermophilic soil bacterium belonging to Actinobacteria. It has been known for its capability to degrade plant cell wall polymers except lignin and pectin. To know whether it can produce enzymes to facilitate lignin degradation, the extracellular proteins bound to sugarcane bagasse were harvested and identified by liquid chromatography tandem mass spectrometry. Among the identified proteins, a putative copper-containing polyphenol oxidase of 241 amino acids, encoded by the locus Tfu_1114, was thought to presumably play a role in lignin degradation. This protein (Tfu1114) was thus expressed in E. coli and characterized. Similarly to common laccases, Tfu1114 is able to catalyze the oxidation reaction of phenolic and nonphenolic lignin related compounds such as 2,6-dimethoxyphenol and veratryl alcohol. More interestingly, it can significantly enhance the enzymatic hydrolysis of bagasse by xylanase and cellulase. Tfu1114 is stable against heat, with a half-life of 4.7 h at 90 °C, and organic solvents. It is sensitive to ethylenediaminetetraacetic acid and reducing agents but resistant to sodium azide, a potent inhibitor of laccases. Atomic absorption spectroscopy indicated that the ratio of copper to the protein monomer is 1, instead of 4, a feature of classical laccases. All these data suggest that Tfu1114 is a novel oxidase with laccase-like activity, potentially useful in biotechnology application.


Subject(s)
Actinomycetales/enzymology , Bacterial Proteins/metabolism , Catechol Oxidase/metabolism , Cellulase/chemistry , Cellulose/chemistry , Endo-1,4-beta Xylanases/chemistry , Saccharum/chemistry , Actinomycetales/chemistry , Actinomycetales/genetics , Amino Acid Sequence , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Biocatalysis , Catechol Oxidase/chemistry , Catechol Oxidase/genetics , Enzyme Stability , Hydrolysis , Industrial Waste/analysis , Kinetics , Lignin/metabolism , Molecular Sequence Data , Molecular Weight , Saccharum/microbiology , Sequence Alignment
15.
Biomed Res Int ; 2013: 732307, 2013.
Article in English | MEDLINE | ID: mdl-24416725

ABSTRACT

The periodic expression and destruction of several cyclins are the most important steps for the exact regulation of cell cycle. Cyclins are degraded by the ubiquitin-proteasome system during cell cycle. Besides, a short sequence near the N-terminal of cyclin B called the destruction box (D-box; CDB) is also required. Fluorescent-protein-based reporter gene system is insensitive to analysis because of the overly stable fluorescent proteins. Therefore, in this study, we use human CDB fused with both enhanced green fluorescent protein (EGFP) at C-terminus and red fluorescent protein (RFP, DsRed) at N-terminus in the transfected human melanoma cells to examine the effects of CDB on different fluorescent proteins. Our results indicated that CDB-fused fluorescent protein can be used to examine the slight gene regulations in the reporter gene system and have the potential to be the system for screening of functional compounds in the future.


Subject(s)
Amino Acid Sequence/genetics , Cell Cycle/genetics , Cyclin B1/chemistry , Genes, Reporter/genetics , Cell Line, Tumor , Cyclin B1/genetics , Cyclin B1/metabolism , Gene Expression Regulation , Green Fluorescent Proteins/chemistry , Green Fluorescent Proteins/genetics , Humans , Luminescent Proteins/chemistry , Luminescent Proteins/genetics , Protein Stability , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Sequence Deletion , Red Fluorescent Protein
16.
Chin J Physiol ; 55(1): 22-30, 2012 Feb 29.
Article in English | MEDLINE | ID: mdl-22242951

ABSTRACT

The purpose of this study was to determine the effects of an extract from Moringa oleifera (MO) on the development of monocrotaline (MCT)-induced pulmonary hypertension (PH) in Wistar rats. An ethanol extraction was performed on dried MO leaves, and HPLC analysis identified niaziridin and niazirin in the extract. PH was induced with a single subcutaneous injection of MCT (60 mg/kg) which resulted in increases in pulmonary arterial blood pressure (Ppa) and in thickening of the pulmonary arterial medial layer in the rats. Three weeks after induction, acute administration of the MO extract to the rats decreased Ppa in a dose-dependent manner that reached statistical significance at a dose of 4.5 mg of freeze-dried extract per kg body weight. The reduction in Ppa suggested that the extract directly relaxed the pulmonary arteries. To assay the effects of chronic administration of the MO extract on PH, control, MCT and MCT+MO groups were designated. Rats in the control group received a saline injection; the MCT and MCT+MO groups received MCT to induce PH. During the third week after MCT treatment, the MCT+MO group received daily i.p. injections of the MO extract (4.5 mg of freeze-dried extract/kg of body weight). Compared to the control group, the MCT group had higher Ppa and thicker medial layers in the pulmonary arteries. Chronic treatments with the MO extract reversed the MCT-induced changes. Additionally, the MCT group had a significant elevation in superoxide dismutase activity when normalized by the MO extract treatments. In conclusion, the MO extract successfully attenuated the development of PH via direct vasodilatation and a potential increase in antioxidant activity.


Subject(s)
Hypertension, Pulmonary/drug therapy , Moringa oleifera/chemistry , Phytotherapy , Plant Extracts/therapeutic use , Superoxide Dismutase/metabolism , Animals , Drug Evaluation, Preclinical , Hypertension, Pulmonary/chemically induced , Lung/enzymology , Male , Monocrotaline , Rats , Rats, Wistar
17.
Int J Mol Sci ; 12(9): 5946-54, 2011.
Article in English | MEDLINE | ID: mdl-22016637

ABSTRACT

The skin-whitening agent, deoxyArbutin, is a potent tyrosinase inhibitor that is safer than hydroquinone and arbutin. However, it is thermolabile in aqueous solutions, where it decomposes to hydroquinone. Pharmaceutical and cosmetic emulsions are normally oil-in-water (o/w) or water-in-oil (w/o) systems; however, emulsions can be formulated with no aqueous phase to produce an anhydrous emulsion system. An anhydrous emulsion system could offer a stable vehicle for compounds that are sensitive to hydrolysis or oxidation. Therefore, to enhance the stability of deoxyArbutin in formulations, we chose the polyol-in-silicone, anhydrous emulsion system as the basic formulation for investigation. The quantity of deoxyArbutin and the accumulation of hydroquinone in both hydrous and anhydrous emulsions at various temperatures were analyzed through an established high performance liquid chromatographic (HPLC) method. The results indicated that water increased the decomposition of deoxyArbutin in the formulations and that the polyol-in-silicone, oil-based, anhydrous emulsion system provided a relatively stable surrounding for the deoxyArbutin that delayed its degradation at 25 °C and 45 °C. Moreover, the composition of the inner hydrophilic phase, containing different amounts of glycerin and propylene glycol, affected the stability of deoxyArbutin. Thus, these results will be beneficial when using deoxyArbutin in cosmetics and medicines in the future.


Subject(s)
Arbutin/analogs & derivatives , Emulsions/chemistry , Polymers/chemistry , Silicon/chemistry , Arbutin/chemistry , Chromatography, High Pressure Liquid , Drug Stability , Hydrolysis , Hydroquinones/chemistry , Oils/chemistry , Temperature , Water/chemistry
18.
Biosci Biotechnol Biochem ; 75(10): 1977-83, 2011.
Article in English | MEDLINE | ID: mdl-21979069

ABSTRACT

Eucalyptus bridgesiana, Cymbopogon martinii, Thymus vulgaris, Lindernia anagallis, and Pelargonium fragrans are five species of herbs used in Asia. Their essential oils were analyzed by GC-MS, and a total of 36 components were detected. The results of our study indicated that, except for the essential oil of P. fragrans, all of the essential oils demonstrated obvious antimicrobial activity against a broad range of microorganisms. The C. martinii essential oil, which is rich in geraniol, was the most effective antimicrobial additive. All of the essential oils demonstrated antioxidant activities on 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay, ß-carotene/linoleic acid assay, and nitric oxide radical scavenging assay. Furthermore, the T. vulgaris essential oil, which possesses plentiful thymol, exhibited the highest antioxidant activity. For P. acnes-induced secretion of pro-inflammatory cytokines, the essential oils of P. aeruginosa, C. martinii, and T. vulgaris reduced the TNF-α, IL-1ß, and IL-8 secretion levels of THP-1 cells.


Subject(s)
Anti-Infective Agents/pharmacology , Anti-Inflammatory Agents/pharmacology , Free Radical Scavengers/pharmacology , Lipoxygenase Inhibitors/pharmacology , Magnoliopsida/chemistry , Oils, Volatile/pharmacology , Anti-Infective Agents/analysis , Anti-Inflammatory Agents/analysis , Arachidonate 5-Lipoxygenase/metabolism , Bacteria/drug effects , Cell Line, Tumor , Cytokines/metabolism , Free Radical Scavengers/analysis , Fungi/drug effects , Humans , Inflammation Mediators/metabolism , Inhibitory Concentration 50 , Lipoxygenase Inhibitors/analysis , Microbial Sensitivity Tests , Oils, Volatile/analysis
19.
Phytomedicine ; 18(14): 1244-9, 2011 Nov 15.
Article in English | MEDLINE | ID: mdl-21802267

ABSTRACT

Citrus fruits are the major source of flavonoids for humans, and flavanones are the main flavonoids in the Citrus species. Among the Citrus flavanones, the glycoside derivatives of naringenin, naringin and narirutin, are the most abundant in grapefruit. The present study aimed to investigate the molecular events of melanogenesis induced by naringenin in murine B16-F10 melanoma cells. Melanin content, tyrosinase activity and Western blot analysis were performed to elucidate the possible underlying mechanisms. Exposure of melanoma cells to naringenin resulted in morphological changes accompanied by the induction of melanocyte differentiation-related markers, such as melanin synthesis, tyrosinase activity, and the expression of tyrosinase and microphthalmia-associated transcription factor (MITF). We also observed an increase in the intracellular accumulation of ß-catenin as well as the phosphorylation of glycogen synthase kinase-3ß (GSK3ß) protein after treatment with naringenin. Moreover, the activity of phosphatidylinositol 3-kinase (PI3K) was up-regulated by naringenin since the phosphorylated level of downstream Akt protein was enhanced. Based on these results, we concluded that naringenin induced melanogenesis through the Wnt-ß-catenin-signalling pathway.


Subject(s)
Citrus/chemistry , Flavanones/pharmacology , Pigmentation/drug effects , Wnt Signaling Pathway , Animals , Blotting, Western , Cell Differentiation , Cell Line, Tumor , Cell Survival , Chromones/pharmacology , Glycogen Synthase Kinase 3/metabolism , Glycogen Synthase Kinase 3 beta , Melanins/metabolism , Melanocytes/drug effects , Melanoma, Experimental/enzymology , Mice , Microphthalmia-Associated Transcription Factor/metabolism , Monophenol Monooxygenase/metabolism , Morpholines/pharmacology , Phosphatidylinositol 3-Kinases/metabolism , Phosphorylation , Up-Regulation , beta Catenin/metabolism
20.
J AOAC Int ; 94(3): 985-90, 2011.
Article in English | MEDLINE | ID: mdl-21797028

ABSTRACT

An online derivatization followed by a disposable electrochemical sensor was used for the determination of arbutin (AR) in cosmetic products. The AR was chemically oxidized by MnO2 and subsequently reduced at inexpensive screen-printed carbon electrodes using a low detection potential which improved the selectivity of the method. The effects of various parameters, such as solution pH, detection potential, and flow rate of the mobile phase, were studied in detail. Under optimal conditions [pH 1.6 (0.1 M H3PO4), detection potential 0.0 V (versus Ag/AgCl), flow rate 0.6 mL/min], the linear range for AR was 0.1-1500 ppm (r2 = 0.999) with LOD of 30.06 ppb (S/N = 3). The practical application of the proposed method was demonstrated by the determination of arbutin concentration in commercial cosmetic products.


Subject(s)
Arbutin/chemistry , Cosmetics/chemistry , Electrochemistry/instrumentation , Electrochemistry/methods , Manganese Compounds/chemistry , Oxidation-Reduction , Oxides/chemistry
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