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*

The epidermal cell-derived thymocyte activating factor (ETAF) produced by keratinocytes and endowed with IL-1 like activities, can potentiate or stimulate a variety of immune reactionsl. In addition to stimulating thymocyte proliferation, it enhanced IL-2 production by mitogen-stimulated lymphocytes. In this study, IL-2 activity stimulated by ETAF was investigated by using peripheral lymphocytes (PBL)and LBRM33 1A5 cell lines. The expression of HLA-DR antigen and the effect of r-IFN-gamma on IL-1 production by keratinocytes were also evaluated : 1. The HLA-DR positive cultured keratinocytes were obser ved in interferon (IFN), interferon-phorbol myristic acid (IFN-PMA) and interferon-lipopolysaccharide (IFN-LPS) groups. There was no HLA-DR expression in the control. PMA. and LPS group. 2. Il-1alpha from supernatants of cultured keratinocytes were produced by all groups including control group. The amounts of IL-1alpha produced by IFN, IFN-PMA and IFN-LPS groups were higher than the other groups. 3. There were no statistical differences between total IL-1alpha from cultured keratinocytes and control or other experimental groups. 4. In experiment for positivie control, the anrounts of IL-2 production by PBL and LBRM33 1A5 cell line were increased proportional to the concentration of added rIL-1. But there was no difference between groups above 1.0 U/ml and 0.5 fmol in LBRM33 1A5 cell line. 5. the production of IL-2 by PBL were increased by phytohemagglutinin (PHA), PHAPMA and PHA-LPS groups. There were no production of IL-2 in control, IFN, LPS, only LPS added group, and IL-1alpha. The IL-2 induced by PHA-PMA group was much higher than the other groups. The production of IL-2 was stimulated by IL-1 of supernatarnts and cell lysates from the cultured keratinocytes, but here was no correlation between IL-2 production and added IL-1. 6. The production of IL-2 by LBRM33 IA5 were increased by PHA, PHA-PMA and PHA-LPS groups. There were no production of IL-2 in the other groups. In summary, the results suggest that supernatants and cell lysates from cultured keratinocytes can stimulate the IL-2 production by human PBL and LBRM33 1A5 cell line. Therefore the ETAF can potentiate or stimulate the immune reactions and enhance IL-2 production by mitogen-stimulated lymphocytes.
Cell Line ; HLA-DR Antigens ; Humans ; Interferons ; Interleukin-1* ; Interleukin-2* ; Keratinocytes* ; Lymphocytes* ; Myristic Acid ; Thymocytes

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

The purposes of this research were to assess dietary fatty acid patterns and to elucidate the relationship between the serum cholesterol levels and dietary fatty acid patterns, plasma fatty acid compositions, BMI (body mass index), and other lipid profile. The subjects were 151 adults aged 23 to 80 years, selected from the Outpatient Clinic and Cardiovascular Department of the Seoul Municipal Hospital. Dietary data were obtained using three day food records. Sixteen dietary fatty acids were analyzed using Korean and US nutrient databases. The subjects were divided into three serum cholesterol levels: desirable (< 200 mg/dl, N = 44), borderline-risk (> or = 200 - < 240 mg/dl, N = 35), and high-risk (> or = 240 mg/dl, N = 72) groups. The high-risk group had higher BMI, waist, and waist to hip ratio (WHR) than the desirable and borderline-risk groups. Serum concentrations of triglyceride, LDL cholesterol and LDL/HDL cholesterol ratio were significantly higher in the high-risk group as compared to those in the other two groups. The serum cholesterol levels were highly correlated with BMI (r = 0.435), triglyceride (r = 0.425) and LDL/HDL cholesterol (r = 0.870) ratio. The highest fatty acid intake was from oleic acid (33 - 34% of total fatty acid intakes), which was followed by linoleic acid (27%), palmitic acid (19%), and stearic acid (7%). There was no correlation between the serum cholesterol levels and the dietary fatty acid intakes, polyunsaturated/monounsaturated/saturated fatty acids (P/M/S) and omega6/omega3 ratios. The correlation between plasma fatty acids such as myristic acid, oleic acid, linoleic acid, and docosahexaenoic acid and serum cholesterol levels was also weak.
Adult* ; Ambulatory Care Facilities ; Cholesterol* ; Cholesterol, LDL ; Fatty Acids* ; Hospitals, Municipal ; Humans ; Linoleic Acid ; Myristic Acid ; Oleic Acid ; Palmitic Acid ; Plasma* ; Seoul ; Triglycerides ; Waist-Hip Ratio

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

PURPOSE: Na+-K+ ATPase Activity has beem estimated by the degree of inhibition of cation transport by cardiac glycosides (ouabain) using Rb-86 as a substrate. The biological characterist-Isc of T1-201 is known to be simiIar to those of potassium as a transport substrate in the presence of glucose, insulin or phobol myristate acetate (PMA). The purpose of this study was to measure ouabain sensitive Na+-K+ ATPase activity using T1-201 and compare with that using Rb-86. MATERIALS AND METHODS: Smooth muscle cells isolated from rat aorta or human placental umbilical artery were cultured, and used to measure cellular Na+-K+ ATPase activity. Na+-K+ ATPase activity was measured as a percentage decrease in cellular uptake of T1-201 or Rb-86 by ouabain under the presence of glucose, insulin or PMA in media. RESULTS: Na+-K+ ATPase ase activity measured with T1-201, as a transport substrate, was not different from those measured with Rb-86 in rat or human smooth muscle cell preparation. Incubation with high concentration glucose resulted in about 30% decrease in enzyme activity. In contrast, insulin or PMA resulted in 50-70% or 28% increases from baseline activity, respectively. CONCLUSION: These results suggests that T1-201 could replace Rb-86 in measurement of ouabain sensititive Na+-K+ ATPase activity in vitro. High level of glucose concentration decreased cellular Na+-K+ ATPase activity, but insulin or PMA increased it.
Adenosine Triphosphatases* ; Animals ; Aorta ; Cardiac Glycosides ; Glucose ; Humans ; Insulin ; Membranes* ; Myocytes, Smooth Muscle ; Myristic Acid ; Ouabain ; Potassium ; Rats ; Umbilical Arteries

BACKGROUND: Kerationcytes make up the vast majority of cells within the epidermis. Recent attention has focused on the role keratinocytes may play in the induction of T cell mediated inflammatory responses in skin, particularily because keratinocytes, when activated by immunologic stimuli, express MHC class llAg and secrete cytokines. But in experimentally induced lichenoid tissue reaction by interferon-nu, MHC class ll Ag was not essential for the enhanced T cell trafficking. There is growing evidence that keratinocyte intercellular adhesion molecule-1 (ICAM-1) expression is involved in the epidermal trafficking of T lymphocytes. OBJECT: To investigate the kinetics of expression of the HLA-DR and ICAM-1 on cultured human keratinocytes by recombinant-interferon-nu(IFN) and phorbol myristate acetate(PMA), and the influence of the HLA-DR and ICAM-1 and the stimulation of peripheral blood mononuclear leukocytes(PBML). RESULTS: 1. 1 U/ml of IFN can induce HLA-DR and ICAM-1 on keratinocytes. Expression of both antigens were increased in a dose and exporsure time dependent fashion. But expression of HLA-DR was less sensitive to IFN than ICAM-1 2. ICAM-1 induction was more rapid than HLA-DR. Keratinocytes expressed HLA-DR 6hours after IFN treatment amd increased rapidly after 12 hours. 3. HLA-DR positive keratinocytes were decreased more rapidly that ICAM-1 positive kerationcytes. 4. Proliferations of PBML were slightly inhibited when cultured with keratinocytes which were treated or not treated with IFN. But IFN treated keratinocytes stimulated the PBML more than untreated keratinocytes. Proliferaton of PBML by IFN treated keratinocytes were inhibited by anti-ICAM monoclonal antibody 5. PMA treated keratinocytes stimulated the PBML more than untreated keratinocytes. Proliferation of PBML by PMA treated keratinocytes was inhibited by anti-ICAM monoclonal antibodies and anti-LFA-1 monoclonal antibodies. CONCLUSION: These results suggest that keratinocytes can express not only HLA-DR but also ICAMP1 may play a important role in initiating immunologic response. Complete clarification of the function of HLA-DR and ICAM-1 positive keratinocytes requires furthe5r study
Antibodies, Monoclonal ; Cytokines ; Epidermis ; HLA-DR Antigens* ; Humans* ; Intercellular Adhesion Molecule-1* ; Keratinocytes* ; Kinetics* ; Myristic Acid ; Skin ; T-Lymphocytes

Paclitaxel (Taxol) is known as effective drug for inhibition of cell cycle encouraging in human cancer cells. This drug named an antimicrotubule agent which simulate the mitotic arrest towards an apoptosis. The influence of phorbol 12 myristate 13 acetate (PMA) activated protein kinase C (PKC) and nitric oxide (NO) on taxol-induced apoptosis, is poorly understood. To investigate the effects of PMA and NO on the signal transduction in taxol-induced apoptosis in C6-glial cells, the viability and caspase-3 activity of C6-glial cells were analyzed. Pretreatement with PKC activatior (PMA) protected taxol-induced cell death in C6-glial cells, by inhibited caspases-3 activity. On the other hand, the taxol-induced apoptosis was highly enhanced by sodium nitroprusside (SNP) and lipopolysaccharide (LPS), as NO activator. These results suggest that PMA strongly blocks the apoptotic effect of taxol, while nitric oxide has no protective effects in the process of toxol-induced apoptosis in C6-glial cells.
Apoptosis ; Caspase 3 ; Cell Cycle ; Cell Death ; Hand ; Humans ; Myristic Acid ; Nitric Oxide* ; Nitroprusside ; Paclitaxel ; Protein Kinase C ; Signal Transduction

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