Chemical investigation of the ethyl acetate extract of Gibberella moniliformis JS1055 endophytic fungus derived from a halophyte, Vitex rotundifolia, led to the isolation of nine compounds including 7-butyl-6,8-dihydroxy-3(R)-pent-11-enylisochroman-1-one (1), 7-butyl-6,8-dihydroxy-3(R)-pentylisochroman-1-one (2), 7-butyl-6,8-dihydroxy-3(R)-pentylisochroman-1-one (3), 5α,8α-epidioxyergosta-6,9(11),22-trien-3-ol (4), ergosterol peroxide (5), tetradecanoic acid (6), 8-O-methylfusarubin (7), nicotinic acid (8) and adenosine (9). They were identified by extensive spectroscopic data analysis including 1D, 2D (¹H-¹H COSY, HSQC, HMBC) NMR, and ESIMS. All the isolates (1
Adenosine ; Ergosterol ; Fungi ; Gibberella ; Moniliformis ; Myristic Acid ; Niacin ; Salt-Tolerant Plants ; Statistics as Topic ; Vitex

Leucocyte extravasation has been known to play an important role in inflammatory reactions including contact dermatitis. Previous studies suggested that CD99 regulates β1 integrin activity and may be a novel therapeutic target molecule for inflammatory diseases. In this study, the effects of CD99-derived peptide, CD99CRIII3, on inflammatory reactions in contact dermatitis mouse model were investigated. CD99CRIII3 decreased β1-integrin activity in human monocytic U937 cells. CD99CRIII3 inhibited the adhesion of U937 monocytes to human umbilical vein endothelial cells and their extravasation through human umbilical vein endothelial cells. CD99CRIII3 reduced inflammation in the phorbol myristate acetate-induced contact dermatitis mice in a dose-dependent manner. These results indicate that CD99CRIII3 suppresses the extravasation of monocytes and inflammatory reactions in the animal model of the contact dermatitis, suggesting that CD99CRIII3 could be a new drug candidate against inflammatory skin diseases.
Animals ; Dermatitis, Contact* ; Human Umbilical Vein Endothelial Cells ; Humans ; Inflammation ; Mice* ; Models, Animal ; Monocytes* ; Myristic Acid ; Skin Diseases ; U937 Cells

Streptomyces avermitilis produces clinically useful drugs such as avermectins and oligomycins. Its genome contains approximately 33 cytochrome P450 genes and they seem to play important roles in the biosynthesis of many secondary metabolites. The SAV_7130 gene from S. avermitilis encodes CYP158A3. The amino acid sequence of this enzyme has high similarity with that of CYP158A2, a biflaviolin synthase from S. coelicolor A3(2). Recombinant S. avermitilis CYP158A3 was heterologously expressed and purified. It exhibited the typical P450 Soret peak at 447 nm in the reduced CO-bound form. Type I binding spectral changes were observed when CYP158A3 was titrated with myristic acid; however, no oxidative product was formed. An analog of flaviolin, 2-hydroxynaphthoquinone (2-OH NQ) displayed similar type I binding upon titration with purified CYP158A3. It underwent an enzymatic reaction forming dimerized product. A homology model of CYP158A3 was superimposed with the structure of CYP158A2, and the majority of structural elements aligned. These results suggest that CYP158A3 might be an orthologue of biflaviolin synthase, catalyzing C-C coupling reactions during pigment biosynthesis in S. avermitilis.
Amino Acid Sequence ; Cytochrome P-450 Enzyme System ; Genome ; Myristic Acid ; Oligomycins ; Streptomyces*

OBJECTIVE: Symptomatic disc degeneration develops from inflammatory reactions in the annulus fibrosus (AF). Although inflammatory mediators during annular inflammation have been studied, the roles of matrix metalloproteinases (MMPs) and their inhibitors have not been fully elucidated. In this study, we evaluated the production of MMPs and tissue inhibitors of metalloproteinase (TIMPs) during annular inflammation using an in vitro co-culture system. We also examined the effect of notochordal cells on annular inflammation. METHODS: Human AF (hAF) pellet was co-cultured for 48 hours with phorbol myristate acetate-stimulated macrophage-like THP-1 cells. hAF pellet and conditioned media (CM) from co-cultured cells were assayed for MMPs, TIMPs, and insulin-like growth factor (IGF)-1 levels using real-time reverse-transcriptase polymerase chain reaction and enzyem-linked immunosorbent assay. To evaluate whether notochordal cells affected MMPs or TIMPs production on annular inflammation, hAF co-cultured with notochordal cells from adult New Zealand White rabbits, were assayed. RESULTS: MMP-1, -3, -9; and TIMP-1 levels were significantly increased in CM of hAF co-cultured with macrophage-like cells compared with hAF alone, whereas TIMP-2 and IGF-1 levels were significantly decreased (p<0.05). After macrophage exposure, hAF produced significantly more MMP-1 and -3 and less TIMP-1 and -2. Interleukin-1beta stimulation enhanced MMP-1 and -3 levels, and significantly diminished TIMP-2 levels. Co-culturing with rabbit notochordal cells did not significantly influence MMPs and TIMPs production or COL1A2 gene expression. CONCLUSION: Our results indicate that macrophage-like cells evoke annular degeneration through the regulation of major degradative enzymes and their inhibitors, produced by hAF, suggesting that the selective regulation of these enzymes provides future targets for symptomatic disc degeneration therapy.
Adult ; Coculture Techniques ; Culture Media, Conditioned ; Gene Expression ; Humans ; Inflammation* ; Insulin-Like Growth Factor I ; Interleukin-1beta ; Intervertebral Disc Degeneration* ; Macrophages ; Matrix Metalloproteinases ; Myristic Acid ; Notochord ; Polymerase Chain Reaction ; Rabbits ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2

BACKGROUND: Malassezia species (spp.) are cutaneous opportunistic pathogens and associated with various dermatological diseases including seborrheic dermatitis, dandruff and atopic dermatitis. Almost all Malassezia spp. are obligatorily lipid-dependent, which might be caused by lack of the myristic acid synthesis. Recent genome analysis of M. restricta and M. globosa suggested that the absence of a gene encoding fatty acid synthesis might be compensated by abundant genes encoding hydrolases, which produce fatty acids, and that lipases and phospholipases may play a role in virulence of the fungus. OBJECTIVE: The current study aimed to investigate the contribution of lipases and phospholipases in virulence of the M. restricta as being the most frequently isolated Malassezia spp. from the human skin. METHODS: Swap samples of two different body sites of at least 18 patients with seborrheic dermatitis were obtained and in vivo expression of lipases and phospholipases of M. restricta was analyzed by the gene specific two-step nested RT-PCR. RESULTS: The results of the current study suggest that majority of the patients display expression of lipase RES_0242. CONCLUSION: These data imply a possible role of lipase in the host environment to produce free fatty acids for the fungus.
Dermatitis, Atopic ; Dermatitis, Seborrheic ; Fatty Acids ; Fatty Acids, Nonesterified ; Fungi ; Genes, vif ; Genome ; Humans ; Hydrolases ; Lipase ; Malassezia ; Myristic Acid ; Phospholipases

BACKGROUND: Malassezia species (spp.) are cutaneous opportunistic pathogens and associated with various dermatological diseases including seborrheic dermatitis, dandruff and atopic dermatitis. Almost all Malassezia spp. are obligatorily lipid-dependent, which might be caused by lack of the myristic acid synthesis. Recent genome analysis of M. restricta and M. globosa suggested that the absence of a gene encoding fatty acid synthesis might be compensated by abundant genes encoding hydrolases, which produce fatty acids, and that lipases and phospholipases may play a role in virulence of the fungus. OBJECTIVE: The current study aimed to investigate the contribution of lipases and phospholipases in virulence of the M. restricta as being the most frequently isolated Malassezia spp. from the human skin. METHODS: Swap samples of two different body sites of at least 18 patients with seborrheic dermatitis were obtained and in vivo expression of lipases and phospholipases of M. restricta was analyzed by the gene specific two-step nested RT-PCR. RESULTS: The results of the current study suggest that majority of the patients display expression of lipase RES_0242. CONCLUSION: These data imply a possible role of lipase in the host environment to produce free fatty acids for the fungus.
Dermatitis, Atopic ; Dermatitis, Seborrheic ; Fatty Acids ; Fatty Acids, Nonesterified ; Fungi ; Genes, vif ; Genome ; Humans ; Hydrolases ; Lipase ; Malassezia ; Myristic Acid ; Phospholipases

Since 1994, several different inactivated rabies vaccines have been used to immunize domestic animals such as dogs, cats, and cattle in South Korea. The Korean Veterinary Authority has conducted safety and efficacy testes of inactivated vaccines using laboratory animals. In this study, we applied a molecular method to investigate the genetic characterization of the rabies virus (RABV) genes in six commercial inactivated rabies vaccines, and determined the efficiency of two extraction reagents (i.e., sodium citrate or isopropyl myristate) to separate the vaccine antigens from the antigen/adjuvant complexes. Six partial nucleocapsid (N: 181 bp) and five partial glycoprotein (G: 306 bp) genes were successfully amplified with specific primer sets, which demonstrated that sodium citrate is more efficient than isopropyl myristate in extracting viral RNA from inactivated gel vaccines. In addition, we identified the viral strain of the vaccine by analyzing the nucleotide sequences of the N and the G genes. The nucleotide similarity of the partial N and G genes ranged from 97.1 to 99.4% and from 91.8 to 100% among rabies vaccine strains, respectively, indicating that each manufacturer used different rabies virus strains to produce their vaccines. The molecular method used in this study could also be used to identify viral strains in other inactivated vaccines.
Animals ; Animals, Domestic ; Animals, Laboratory ; Base Sequence ; Cats ; Cattle ; Citrates ; Citric Acid ; Dogs ; Glycoproteins ; Indicators and Reagents ; Myristates ; Myristic Acid ; Nucleocapsid ; Rabies ; Rabies Vaccines ; Rabies virus ; Republic of Korea ; RNA, Viral ; Sodium ; Sprains and Strains ; Testis ; Vaccines ; Vaccines, Inactivated

BACKGROUND: Based on the assertion that apocynin diminishes acute lung injury (ALI) by inhibition of NADPH oxidase, the effect of apocynin was tested in interleukin-1alpha (IL-1)-induced ALI in rats. METHODS: IL-1 was insufflated into the trachea of Sprague-Dawley rats to induce ALI, and apocynin (8 mg/kg) was given intravenously for inhibition of NADPH oxidase. In addition, we determined whether apocynin inhibited generation of superoxide anions from isolated human neutrophils. Five hours after IL-1 instillation, lung injury parameters, expression of cytosolic phospholipase A2 (cPLA2) by cells from bronchoalveolar lavage (BAL), an index of oxidative stress in lung tissues (gamma-glutamyltranspeptidase, activity), and ultrastructure of alveolar type II (AT II) cells were evaluated. RESULTS: Apocynin decreased the generation of free radicals from phorbol myristate (PMA)-activated neutrophils in vitro, but did not ameliorate ALI. IL-1 induced enhancement of the expression of cPLA2 on neutrophils was not altered by apocynin. CONCLUSION: Apocynin induced suppression of the generation of superoxide anions from neutrophils by inhibition of NADPH oxidase does not attenuate IL-1-induced ALI in rats.
Acetophenones ; Acute Lung Injury ; Animals ; Bronchoalveolar Lavage ; Cytosol ; Free Radicals ; Humans ; Interleukin-1 ; Interleukin-1alpha ; Lung ; Lung Injury ; Myristic Acid ; NADPH Oxidase ; Neutrophils ; Oxidative Stress ; Phorbols ; Phospholipases A2 ; Rats ; Rats, Sprague-Dawley ; Superoxides ; Trachea

Fatty acid binding and oxidation kinetics for wild type P450(BM3) (CYP102A1) from Bacillus megaterium have been found to display chain length-dependent homotropic behavior. Laurate and 13-methyl-myristate display Michaelis-Menten behavior while there are slight deviations with myristate at low ionic strengths. Palmitate shows Michaelis-Menten kinetics and hyperbolic binding behavior in 100 mmol/L phosphate, pH 7.4, but sigmoidal kinetics (with an apparent intercept) in low ionic strength buffers and at physiological phosphate concentrations. In low ionic strength buffers both the heme domain and the full-length enzyme show complex palmitate binding behavior that indicates a minimum of four fatty acid binding sites, with high cooperativity for the binding of the fourth palmitate molecule, and the full-length enzyme showing tighter palmitate binding than the heme domain. The first flavin-to-heme electron transfer is faster for laurate, myristate and palmitate in 100 mmol/L phosphate than in 50 mmol/L Tris (pH 7.4), yet each substrate induces similar high-spin heme content. For palmitate in low phosphate buffer concentrations, the rate constant of the first electron transfer is much larger than k (cat). The results suggest that phosphate has a specific effect in promoting the first electron transfer step, and that P450(BM3) could modulate Bacillus membrane morphology and fluidity via palmitate oxidation in response to the external phosphate concentration.
Bacterial Proteins ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Fatty Acids ; chemistry ; metabolism ; Lauric Acids ; chemistry ; metabolism ; Myristic Acid ; chemistry ; metabolism ; NADPH-Ferrihemoprotein Reductase ; metabolism ; Osmolar Concentration ; Oxidation-Reduction ; Palmitic Acid ; chemistry ; metabolism ; Structure-Activity Relationship

Silmazine(R) cream is an antibiotic agent widely used in burn therapy. It consists of Propylene glycol, Stearyl alcohol, Isopropyl Myristate, Sorbitan mono-oleate, Methyl-p-hydroxybenzoate, Polyoxyl 40 stearate and varseline. A 24-year- old female presented with well-demarcated erythematous papules and vesicles with an itching sensation on the dorsal area of her right hand. She had applied Silmazine(R) cream on the dorsal area of her right handfor 4 days and the skin lesion became aggravated. A patch test with Silmazine(R) cream 'as is' showed a positive reaction and propylene glycol and stearyl alcohol, ingredients in Silmazine(R) cream, revealed a positive reaction. These two agents are known as weak sensitizers that can produce allergic contact dermatitis. There are some reports of allergic contact dermatitis from propylene glycol and stearyl alcohol used topically. As far as we know, there are no reports of allergic contact dermatitis from propylene glycol and stearyl alcohol in the Silmazine(R) cream (Silver sulfadiazine) that is commonly used as topical antibiotic medication for burns. We report this rare case of allergic contact dermatitis from propylene glycol and stearyl alcohol in Silmazine(R) cream (Silver sulfadiazine).
2-Propanol ; Alkenes ; Burns ; Dermatitis, Allergic Contact ; Fatty Alcohols ; Female ; Hand ; Humans ; Myristates ; Myristic Acid ; Patch Tests ; Propylene Glycol ; Pruritus ; Sensation ; Skin