Sample records for psoralen photo-crosslinked mrna-puromycin

  1. Photochemical Acceleration of DNA Strand Displacement by Using Ultrafast DNA Photo-crosslinking. (United States)

    Nakamura, Shigetaka; Hashimoto, Hirokazu; Kobayashi, Satoshi; Fujimoto, Kenzo


    DNA strand displacement is an essential reaction in genetic recombination, biological processes, and DNA nanotechnology. In particular, various DNA nanodevices enable complicated calculations. However, it takes time before the output is obtained, so acceleration of DNA strand displacement is required for a rapid-response DNA nanodevice. Herein, DNA strand displacement by using DNA photo-crosslinking to accelerate this displacement is evaluated. The DNA photo-crosslinking of 3-cyanovinylcarbazole ( CNV K) was accelerated at least 20 times, showing a faster DNA strand displacement. The rate of photo-crosslinking is a key factor and the rate of DNA strand displacement is accelerated through ultrafast photo-crosslinking. The rate of DNA strand displacement was regulated by photoirradiation energy. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Hypertrichosis by Topical Psoralen

    Directory of Open Access Journals (Sweden)

    K Pavithran


    Full Text Available Two cases in which topical psoralen induced hypertrichosis in the depigmented patches of vitiligo are being reported. -The possible mechanisms for this hypertrichosis are briefly discussed.

  3. Photo-Crosslinked Biodegradable Hydrogels Prepared From Fumaric Acid Monoethyl Ester-Functionalized Oligomers for Protein Delivery

    NARCIS (Netherlands)

    Jansen, Janine; Mihov, George; Feijen, Jan; Grijpma, Dirk W.

    Photo-crosslinkable, fumaric acid monoethyl ester-functionalized triblock oligomers are synthesized and copolymerized with N-vinyl-2-pyrrolidone to form biodegradable photo-crosslinked hydrogels. Poly(ethylene glycol) is used as the middle hydrophilic segment and the hydrophobic segments are based

  4. Preparation and mechanical properties of photo-crosslinked poly(trimethylene carbonate) and nano-hydroxyapatite composites

    NARCIS (Netherlands)

    Geven, Mike Alexander; Barbieri, D.; Yuan, Huipin; de Bruijn, Joost Dick; Grijpma, Dirk W.


    Composite materials of photo-crosslinked poly(trimethylene carbonate) and nanoscale hydroxyapatite were prepared and their mechanical characteristics for application as orbital floor implants were assessed. The composites were prepared by solvent casting poly(trimethylene carbonate) macromers with

  5. Photo-crosslinkable hydrogel-based 3D microfluidic culture device. (United States)

    Lee, Youlee; Lee, Jong Min; Bae, Pan-Kee; Chung, Il Yup; Chung, Bong Hyun; Chung, Bong Geun


    We developed the photo-crosslinkable hydrogel-based 3D microfluidic device to culture neural stem cells (NSCs) and tumors. The photo-crosslinkable gelatin methacrylate (GelMA) polymer was used as a physical barrier in the microfluidic device and collagen type I gel was employed to culture NSCs in a 3D manner. We demonstrated that the pore size was inversely proportional to concentrations of GelMA hydrogels, showing the pore sizes of 5 and 25 w/v% GelMA hydrogels were 34 and 4 μm, respectively. It also revealed that the morphology of pores in 5 w/v% GelMA hydrogels was elliptical shape, whereas we observed circular-shaped pores in 25 w/v% GelMA hydrogels. To culture NSCs and tumors in the 3D microfluidic device, we investigated the molecular diffusion properties across GelMA hydrogels, indicating that 25 w/v% GelMA hydrogels inhibited the molecular diffusion for 6 days in the 3D microfluidic device. In contrast, the chemicals were diffused in 5 w/v% GelMA hydrogels. Finally, we cultured NSCs and tumors in the hydrogel-based 3D microfluidic device, showing that 53-75% NSCs differentiated into neurons, while tumors were cultured in the collagen gels. Therefore, this photo-crosslinkable hydrogel-based 3D microfluidic culture device could be a potentially powerful tool for regenerative tissue engineering applications. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Programming reversibly self-folding origami with micropatterned photo-crosslinkable polymer trilayers. (United States)

    Na, Jun-Hee; Evans, Arthur A; Bae, Jinhye; Chiappelli, Maria C; Santangelo, Christian D; Lang, Robert J; Hull, Thomas C; Hayward, Ryan C


    Self-folding microscale origami patterns are demonstrated in polymer films with control over mountain/valley assignments and fold angles using trilayers of photo-crosslinkable copolymers with a temperature-sensitive hydrogel as the middle layer. The characteristic size scale of the folds W = 30 μm and figure of merit A/ W (2) ≈ 5000, demonstrated here represent substantial advances in the fabrication of self-folding origami. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Photo-crosslinked HAMA hydrogel with cordycepin encapsulated chitosan microspheres for osteoarthritis treatment. (United States)

    Xia, Chen; Chen, Pengfei; Mei, Sheng; Ning, Lei; Lei, Chenyang; Wang, Jiying; Zhang, Jianfeng; Ma, Jianjun; Fan, Shunwu


    Autophagy is a protective mechanism in normal cartilage. The present study aimed to investigate the synergistic therapeutic effect of promotion of chondrocyte autophagy via exposure to cordycepin encapsulated by chitosan microspheres (CM-cordycepin) and photo-crosslinked hyaluronic acid methacrylate (HAMA) hydrogel, with the goal of evaluating CM-cordycepin as a treatment for patients with osteoarthritis. First, we developed and evaluated the characteristics of HAMA hydrogels and chitosan microspheres. Next, we measured the effect of cordycepin on cartilage matrix degradation induced by IL1-β in chondrocytes and an ex vivo model. Cordycepin protects cartilage from degradation partly by activation of autophagy. Moreover, we surgically induced osteoarthritis in mice, which were injected intra-articularly with CM-cordycepin and HAMA. The combination of CM-cordycepin and HAMA hydrogel retarded the progression of surgically induced OA. Cordycepin ameliorated cartilage matrix degradation at least partially by inducing autophagy in vivo. Our results demonstrate that the combination of cordycepin encapsulated by CMs and photo-crosslinked HAMA hydrogel could be a promising strategy for treating patients with osteoarthritis.

  8. Formation of interfacial network structure via photo-crosslinking in carbon fiber/epoxy composites

    Directory of Open Access Journals (Sweden)

    S. H. Deng


    Full Text Available A series of diblock copolymers (poly(n-butylacrylate-co-poly(2-hydroxyethyl acrylate-b-poly(glycidyl methacrylate ((PnBA-co-PHEA-b-PGMA, containing a random copolymer block PnBA-co-PHEA, were successfully synthesized by atom transfer radical polymerization (ATRP. After being chemically grafted onto carbon fibers, the photosensitive methacrylic groups were introduced into the random copolymer, giving a series of copolymers (poly(n-butylacrylate-co-poly(2-methacryloyloxyethyl acrylate-b-poly(glycidyl methacrylate((PnBA-co-PMEA-b-PGMA. Dynamic mechanical analysis indicated that the random copolymer block after ultraviolet (UV irradiation was a lightly crosslinked polymer and acted as an elastomer, forming a photo-crosslinked network structure at the interface of carbon fiber/epoxy composites. Microbond test showed that such an interfacial network structure greatly improved the cohesive strength and effectively controlled the deformation ability of the flexible interlayer. Furthermore, three kinds of interfacial network structures, i physical crosslinking by H-bonds, ii chemical crosslinking by photopolymerization, and iii interpenetrating crosslinked network by photopolymerization and epoxy curing reaction were received in carbon fiber/epoxy composite, depending on the various preparation processes.

  9. Preparation and mechanical properties of photo-crosslinked poly(trimethylene carbonate) and nano-hydroxyapatite composites. (United States)

    Geven, Mike A; Barbieri, Davide; Yuan, Huipin; de Bruijn, Joost D; Grijpma, Dirk W


    Composite materials of photo-crosslinked poly(trimethylene carbonate) and nanoscale hydroxyapatite were prepared and their mechanical characteristics for application as orbital floor implants were assessed. The composites were prepared by solvent casting poly(trimethylene carbonate) macromers with varying amounts of nano-hydroxyapatite and subsequent photo-crosslinking. The incorporation of the nano-hydroxyapatite into the composites was examined by thermogravimetric analysis, scanning electron microscopy and gel content measurements. The mechanical properties were investigated by tensile testing and trouser tearing experiments. Our results show that nano-hydroxyapatite particles can readily be incorporated into photo-crosslinked poly(trimethylene carbonate) networks. Compared to the networks without nano-hydroxyapatite, incorporation of 36.3 wt.% of the apatite resulted in an increase of the E modulus, yield strength and tensile strength from 2.2 MPa to 51 MPa, 0.5 to 1.4 N/mm2 and from 1.3 to 3.9 N/mm2, respectively. We found that composites containing 12.4 wt.% nano-hydroxyapatite had the highest values of strain at break, toughness and average tear propagation strength (376% , 777 N/mm2 and 3.1 N/mm2, respectively).

  10. Psoralen-ultraviolet A treatment with Psoralen-ultraviolet B therapy in the treatment of psoriasis. (United States)

    Ahmed Asim, Sadaf; Ahmed, Sitwat; Us-Sehar, Najam


    To compare the conventional psoralen-ultraviolet A treatment with psoralen-ultraviolet B therapy in the treatment of psoriasis. We studied 50 patients of plaque type psoriasis who were selected to receive either conventional psoralen-ultraviolet A or psoralen-ultraviolet B treatment. There was no significant difference between the two treatment groups in the number of patients whose skin cleared of psoriasis or the number of exposures required for clearance. Profile of side effects and disease status was also similar after three months of follow up. Psoralen-ultraviolet B treatment is as effective as conventional psoralen-ultraviolet A in the treatment of psoriasis. Further long term studies are needed to assess the safety of psoralen-ultraviolet B.

  11. Photo-Crosslinked Poly(ε-caprolactone fumarate) Networks for Peripheral Nerve Regeneration: Physical Properties and Preliminary Biological Evaluations (United States)

    Wang, Shanfeng; Yaszemski, Michael J.; Knight, Andrew M.; Gruetzmacher, James A.; Windebank, Anthony J.; Lu, Lichun


    In an effort of achieving suitable biomaterials for peripheral nerve regeneration, we present a material design strategy of combining a crystallite-based physical network and a crosslink-based chemical network. Biodegradable polymer disks and conduits have been fabricated by photo-crosslinking three poly(ε-caprolactone fumarate)s (PCLF530, PCLF1250, and PCLF2000), which were synthesized from the precursor poly(ε-caprolactone) (PCL) diols with nominal molecular weights of 530, 1250, and 2000 g.mol−1, respectively. Thermal properties such as glass transition temperature (Tg), melting temperature (Tm), and crystallinity of photo-crosslinked PCLFs were examined and correlated with their rheological and mechanical properties. Furthermore, in vitro degradation of uncrosslinked and crosslinked PCLFs in PBS crosslinked PCLFs in 1 N NaOH aqueous solution at 37 °C was studied. In vitro cytocompatibility, attachment, and proliferation of Schwann cell precursor line SPL201 cells on three PCLF networks were investigated. Crosslinked PCLF2000 with the highest crystallinity and mechanical properties was found to best support cell attachment and proliferation. Using a new photo-crosslinking method, single-lumen crosslinked PCLF nerve conduits without defects were fabricated in a glass mold. Crosslinked PCLF2000 nerve conduits were selected for evaluation in a 1-cm gap rat sciatic nerve model. Histological evaluation demonstrated that the material was biocompatible with sufficient strength to hold sutures in place after 6 and 17 weeks of implantation. Nerve cable with myelinated axons was found in the crosslinked PCLF2000 nerve conduit. PMID:19171506

  12. High-Resolution N6-Methyladenosine (m6A) Map Using Photo-Crosslinking-Assisted m6A Sequencing**


    Chen, Kai; Lu, Zhike; Wang, Xiao; Fu, Ye; Luo, Guan-Zheng; Liu, Nian; Han, Dali; Dominissini, Dan; Dai, Qing; Pan, Tao; He, Chuan


    N6-methyladenosine (m6A) is an abundant internal modification in eukaryotic mRNA and plays regulatory roles in mRNA metabolism. However, methods to precisely locate the m6A modification remain limited. We present here a photo-crosslinking-assisted m6A sequencing strategy (PA-m6A-seq) to more accurately define sites with m6A modification. Using this strategy, we obtained a high-resolution map of m6A in a human transcriptome. The map resembles the general distribution patte...

  13. Preserved bioactivity and tunable release of a SDF1-GPVI bi-specific protein using photo-crosslinked PEGda hydrogels. (United States)

    Schesny, Marianne K; Monaghan, Michael; Bindermann, Andrea H; Freund, Désirée; Seifert, Martina; Eble, Johannes A; Vogel, Sebastian; Gawaz, Meinrad P; Hinderer, Svenja; Schenke-Layland, Katja


    Chemokine-induced stem cell recruitment is a promising strategy for post myocardial infarction treatment. Injection of stromal cell-derived factor 1 (SDF1) has been shown to attract bone marrow-derived progenitor cells (BMPCs) from the blood that have the potential to differentiate into cardiovascular cells, which support angiogenesis, enabling the improvement of myocardial function. SDF1-GPVI bi-specific protein contains a glycoprotein VI (GPVI)-domain that serves as an anchor for collagen type I (Col I) and III, which are exposed in the wall of injured vasculature. In this study, we generated a cytocompatible hydrogel via photo-crosslinking of poly(ethylene glycol) diacrylate that serves as a reservoir for SDF1-GPVI. Controlled and sustained release of SDF1-GPVI was demonstrated over a period of 7 days. Release features were modifiable depending on the degree of the crosslinking density. Functionality of the GPVI-domain was investigated using a GPVI-binding ELISA to Col I. Activity of the SDF1-domain was tested for its CXCR4 binding potential. Preserved functionality of SDF1-GPVI bi-specific protein after photo-crosslinking and controllable release was successfully demonstrated in vitro supporting the implementation of this drug delivery system as a powerful tool for therapeutic protein delivery in the treatment of cardiovascular ischemic disease. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  14. Fabrication of poly(propylene fumarate)-based orthopaedic implants by photo-crosslinking through transparent silicone molds. (United States)

    Timmer, Mark D; Carter, Cory; Ambrose, Catherine G; Mikos, Antonios G


    This work presents a new molding process for photo-crosslinked, degradable polymeric networks of poly(propylene fumarate) (PPF) and the crosslinking agent poly(propylene fumarate)-diacrylate (PPF-DA). Transparent room temperature vulcanizing silicone molds were fabricated for parts ranging from simple test coupons to orthopaedic implants. The PPF/PPF-DA resin blend was injected into the cavity and photo-crosslinked as light was transmitted through the mold wall. The volumetric shrinkage, mechanical properties, and the effects of gamma sterilization were reported for molded PPF/PPF-DA networks prepared with varying compositions of the two polymer components. The shrinkage decreased while the mechanical properties displayed a general increasing trend when more of the crosslinking agent was incorporated into the network. Gamma irradiation resulted in an improvement of the mechanical properties. In addition, PPF/PPF-DA replicates of a 70:30 poly(L/DL-lactide) biodegradable fixation plate and a bone allograft interbody fusion spacer were produced to evaluate the performance of PPF/PPF-DA as an orthopaedic implant and allow for a comparison to be made with materials that have been established for clinical use.

  15. The amelioration of cartilage degeneration by photo-crosslinked GelHA hydrogel and crizotinib encapsulated chitosan microspheres. (United States)

    Chen, Pengfei; Mei, Sheng; Xia, Chen; Zhu, Ren; Pang, Yichuan; Wang, Jiying; Zhang, Jianfeng; Shao, Fangchun; Fan, Shunwu


    The present study aimed to investigate the synergistic therapeutic effect of decreaseing cartilage angiogenesis via exposure to crizotinib encapsulated by chitosan microspheres and photo-crosslinked hydrogel, with the goal of evaluating crizotinib as a treatment for osteoarthritis. First, we developed and evaluated the characteristics of hydrogels and chitosan microspheres. Next, we measured the effect of crizotinib on the cartilage degeneration induced by interleukin-1β in chondrocytes. Crizotinib ameliorated the pathological changes induced by interleukin-1β via its anti-angiogenesis function. In addition, we surgically induced osteoarthritis in mice, which were then injected intra-articularly with crizotinib-loaded biomaterials. Cartilage matrix degradation, expression of vascular endothelial growth factor and extracellular signal-regulated kinases 1/2 were evaluated after surgery. Treatment with the combination of crizotinib-loaded biomaterials retarded the progression of surgically induced osteoarthritis. Crizotinib ameliorated cartilage matrix degradation by promoting anti-angiogenesis and impeding extracellular signal-regulated kinases 1/2 signaling pathway. Our results demonstrate that the combination of photo-crosslinked hydrogel and crizotinib-loaded chitosan microspheres might represent a promising strategy for osteoarthritis treatment.

  16. Resilient Amorphous Networks Prepared by Photo-Crosslinking High-Molecular-Weight D,L-Lactide and Trimethylene Carbonate Macromers : Mechanical Properties and Shape-Memory Behavior

    NARCIS (Netherlands)

    Sharifi, Shahriar; Grijpma, Dirk W.


    Tough networks are prepared by photo-crosslinking high-molecular-weight DLLA and TMC macromers. These amorphous networks exhibit tunable thermal and mechanical properties and have excellent shape-memory features. Variation of the monomer ratio allows adjustment of Tg between approximately -13 and

  17. Resilient Amorphous Networks Prepared by Photo-Crosslinking High-Molecular-Weight D,L-Lactide and Trimethylene Carbonate Macromers: Mechanical Properties and Shape-Memory Behavior

    NARCIS (Netherlands)

    Sharifi, Shahriar; Grijpma, Dirk W.


    Tough networks are prepared by photo-crosslinking high-molecular-weight DLLA and TMC macromers. These amorphous networks exhibit tunable thermal and mechanical properties and have excellent shape-memory features. Variation of the monomer ratio allows adjustment of Tg between approximately −13 and

  18. Reaction of the psoralens with deoxyribonucleic acid

    Energy Technology Data Exchange (ETDEWEB)

    Hearst, J.E.; Isaacs, S.T.; Kanne, D.; Rapoport, H.; Straub, K.


    This review has been limited to DNA reactions, and methods for radiolabelling psoralens, which have played an essential role in the isolation and purification of the photoproducts of the reaction between psoralens and nucleic acids, are reviewed herein. High-performance liquid chromatography procedures used for the separation of the photoadducts either as the nucleic adducts or as the free base adducts are discussed. Conclusions drawn from high-resolution proton magnetic resonance and high-resolution mass spectrometric studies of the structural determination of the isolated and purified adducts presented here have led to the assignment of the stereochemistry of the products and leave no doubt that the DNA helix influences both the kinetics and the stereospecificity of the photochemical reaction. The optical properties of the reactants and the photoproducts are catalogued. The kinetics of the photochemistry and its relationship to the fundamental properties of the various psoralen derivatives are discussed. 41 references, 19 figures, 8 tables.

  19. Synthesis of photo-crosslinkable hyaluronan with tailored degree of substitution suitable for production of water resistant nanofibers. (United States)

    Huerta-Angeles, Gloria; Brandejsová, Martina; Knotková, Kateřina; Hermannová, Martina; Moravcová, Martina; Šmejkalová, Daniela; Velebný, Vladimír


    In this work, hyaluronan (HA) was grafted by a novel and an efficient mixed anhydrides methodology with (hetero)-aryl and aliphatic acrylic moieties suitable for cross-linking. A precise control of stoichiometry was achieved. Derivatives with degree of substitution (DS) below 20% did not show self-crosslinking. Due to mild reaction conditions, a negligible degradation of the polysaccharide was obtained. The influence of the feed components on the reaction efficiency and DS were studied up to 200 g/batch. The structure of the modified HA was characterized by Infrared Spectroscopy, Nuclear Magnetic Resonance, SEC-MALS and chromatographic analyses. Enzymatic degradation of derivatives was performed and isolated dimers demonstrated to be non-cytotoxic. The feasibility of the grafted HA for electrospinning with subsequent photo-crosslinking to avoid nanofibers water dissolution was demonstrated. The biocompatibility of the material, its degradation products, and the formation of honeycomb porous structures also proved the potential of the material for future in vivo applications. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Photochemistry of psoralen-DNA adducts, biological effects of psoralen-DNA adducts, applications of psoralen-DNA photochemistry

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Yun-bo


    This thesis consists of three main parts and totally eight chapters. In Part I, The author will present studies on the photochemistry of psoralen-DNA adducts, specifically, the wavelength dependencies for the photoreversals of thymidine-HMT (4'-hydroxymethyl-4, 5', 8-trimenthylpsoralen) monoadducts and diadduct and the same adducts incorporated in DNA helices and the wavelength dependecies for the photocrossslinking of thymidine-HMT monoadducts in double-stranded helices. In Part II, The author will report some biological effects of psoralen-DNA adducts, i.e., the effects on double-stranded DNA stability, DNA structure, and transcription by E. coli and T7 RNA polymerases. Finally, The author will focus on the applications of psoralen-DNA photochemistry to investigation of protein-DNA interaction during transcription, which includes the interaction of E. coli and T7 RNA polymerases with DNA in elongation complexes arrested at specific psoralen-DNA adduct sites as revealed by DNase I footprinting experiments. 123 refs., 52 figs., 12 tabs.

  1. Continuous Preparation of Hollow Polymeric Nanocapsules Using Self-Assembly and a Photo-Crosslinking Process of an Amphiphilic Block Copolymer

    Directory of Open Access Journals (Sweden)

    Xuan Don Nguyen


    Full Text Available This paper presents a fabrication method of hollow polymeric nanocapsules (HPNCs. The HPNCs were examined to reduce light trapping in an organic light emitting diodes (OLED device by increasing the refractive index contrast. They were continuously fabricated by the sequential process of self-assembly and photo-crosslinking of an amphiphilic block copolymer of SBR-b-PEGMA, poly(styrene-r-butadiene-b-poly(poly(ethylene glycol methyl ether methacrylate in a flow-focusing microfluidic device. After the photo-crosslinking process, the produced HPNCs have a higher resistance to water and organic solvents, which is applicable to the fabrication process of optical devices. The morphology and hollow structure of the produced nanocapsules were determined by transmission electron microscopy (TEM, scanning electron microscopy (SEM, and atomic force microscopy (AFM. Also, their size control was examined by varying the ratio of inlet flow rates and the morphological difference was studied by changing the polymer concentration. The size was measured by dynamic light scattering (DLS. The refractive index of the layer with and without the HPNCs was measured, and a lower refractive index was obtained in the HPNCs-dispersed layer. In future work, the light extraction efficiency of the HPNCs-dispersed OLED will be examined.

  2. Psoralens and coumarins for receptor targeting on epidermal cells

    Energy Technology Data Exchange (ETDEWEB)

    Jetter, M.M.


    Specific binding sites have been identified for the psoralens, discrete from DNA, in different epidermal cell lines. These receptors are saturable and are alkylated by the action of psoralens + UVA light. A psoralen receptor has been partially purified and established to be a protein of approximately 20,000 daltons. Inhibition of the binding of epidermal growth factor to its receptor and inhibition of the tyrosine kinase activity of the EGF receptor has been associated with PUVA treatment. These findings conflict with the general assumption that the biological effects of psoralens as photoactive compounds are associated with their ability to covalently bind to and crosslink DNA. In collaboration with Laskin's,laboratory, several classes of psoralen agonists were synthesized. These compounds include coumarins, furocoumarin and benzodipyran-2-one derivatives. The methods of preparation were varied and include variants of the Claisen rearrangement, acid and base-catalyzed condensations. The synthesized compounds were tested for their potential inhibition of {sup 125}I-EGF receptor binding. It was discovered that many of these agents showed potent inhibition activity similar to the psoralens. This data offers the possibility that sites of action, other than DNA, are involved in the mechanism by which photoactivated psoralens modulate epidermal cell lines.

  3. X-Ray Psoralen Activated Cancer Therapy (X-PACT.

    Directory of Open Access Journals (Sweden)

    Mark Oldham

    Full Text Available This work investigates X-PACT (X-ray Psoralen Activated Cancer Therapy: a new approach for the treatment of solid cancer. X-PACT utilizes psoralen, a potent anti-cancer therapeutic with current application to proliferative disease and extracorporeal photopheresis (ECP of cutaneous T Cell Lymphoma. An immunogenic role for light-activated psoralen has been reported, contributing to long-term clinical responses. Psoralen therapies have to-date been limited to superficial or extracorporeal scenarios due to the requirement for psoralen activation by UVA light, which has limited penetration in tissue. X-PACT solves this challenge by activating psoralen with UV light emitted from novel non-tethered phosphors (co-incubated with psoralen that absorb x-rays and re-radiate (phosphoresce at UV wavelengths. The efficacy of X-PACT was evaluated in both in-vitro and in-vivo settings. In-vitro studies utilized breast (4T1, glioma (CT2A and sarcoma (KP-B cell lines. Cells were exposed to X-PACT treatments where the concentrations of drug (psoralen and phosphor and radiation parameters (energy, dose, and dose rate were varied. Efficacy was evaluated primarily using flow cell cytometry in combination with complimentary assays, and the in-vivo mouse study. In an in-vitro study, we show that X-PACT induces significant tumor cell apoptosis and cytotoxicity, unlike psoralen or phosphor alone (p<0.0001. We also show that apoptosis increases as doses of phosphor, psoralen, or radiation increase. Finally, in an in-vivo pilot study of BALBc mice with syngeneic 4T1 tumors, we show that the rate of tumor growth is slower with X-PACT than with saline or AMT + X-ray (p<0.0001. Overall these studies demonstrate a potential therapeutic effect for X-PACT, and provide a foundation and rationale for future studies. In summary, X-PACT represents a novel treatment approach in which well-tolerated low doses of x-ray radiation are delivered to a specific tumor site to generate UVA

  4. Isolation and purification of psoralen and isopsoralen and their ...

    African Journals Online (AJOL)

    Nude rat model of osteosarcoma was established; the rats were randomized into: normal saline group, psoralen low- and high-dose groups, isopsoralen low- and high-dose groups, and cisplatin group. Osteosarcoma volume and weight inhibition rates in nude rats in each group were observed; radioimmunoassay was ...

  5. Isolation and purification of psoralen and isopsoralen and their ...

    African Journals Online (AJOL)

    into: normal saline group, psoralen low- and high-dose groups, isopsoralen low- and high-dose groups, and cisplatin group. Osteosarcoma volume and weight inhibition rates in nude rats in each group were observed; radioimmunoassay was used to determine the serum alkaline phosphatase activity; peripheral blood cell ...

  6. Isolation and purification of psoralen and isopsoralen and their ...

    African Journals Online (AJOL)

    Isolation and purification of psoralen and isopsoralen and their efficacy and safety in the treatment of osteosarcoma in nude rats. Honghui Lu¥ , Lihai Zhang¥, Daohong Liu, Peifu Tang2, Feixiang Song. Orthopaedic Department , People's Liberation Army General Hospital, Beijing 100000, China. ¥Authors Contributed ...

  7. Isolation and characterization of pyrimidine-psoralen-pyrimidine photodiadducts from DNA. [Ultraviolet radiation

    Energy Technology Data Exchange (ETDEWEB)

    Kanne, D.; Straub, K.; Hearst, J.E.; Rapoport, H.


    The isolation and characterization of pyrimidine-psoralen-pyrimidine photodiadducts from DNA are reported for the first time. For each of the four psoralens studied, a single pair of diastereomeric thymidine-psoralen-thymidine photodiadducts, each with cis-syn stereochemistry, was found to account for > 90% of the diadducts formed. Additionally, pulse-chase experiments that establish that these photo cross-links are formed by cycloaddition of a second thymidine residue to the 3,4 double bond (pyrone side) of an initially formed 4',5' (furan-side) psoralen-thymidine photomonoadduct have been carried out.

  8. Cavamax W7 composite psoralen ethosomal gel versus cavamax W7 psoralen solid complex gel for topical delivery: A comparative evaluation (United States)

    Kumari, Smriti; Pathak, Kamla


    Aim: The present research work was aimed to formulate and characterize psoralen-encapsulated cavamax W7 composite ethosomal gel and compare its in vitro and ex vivo behavior against psoralen-cavamax W7-complex reference gel. Materials and Methods: A total of nine formulations of composite ethosomes were prepared by injection method using 32 factorial design and entrapment efficiency was designated as dependent variable. Concomitantly, psoralen was complexed with cavamax W7 (1:1 molar ratio) by kneading method and formation of complex was confirmed by Diffuse reflectance spectroscopy (DRS), scanning electron microscopy (SEM), X-ray diffraction (XRD), and differential scanning calorimetry (DSC). Results: F9 with vesicle size of 183 ± 2.8 nm, and highest % entrapment efficiency of 98.12 ± 1.15 was selected as optimized formulation. Transmission electron microscopy (TEM) revealed uniform and spherical shaped vesicles. The optimized formulation F9 was formulated as carbapol gel and compared against ethosomal gel, psoralen gel, and psoralen cavamax W7 complex gel. The gels were evaluated for permeation characteristics and the rank order was composite ethosomal gel > ethosomal gel > psoralen-cavamax W7 complex gel > psoralen gel. The ethosomal gel (G5) with highest in vitro permeation of 82.48 ± 2.23% was subjected to in vivo Confocal laser scanning microscopy (CLSM) studies using rhodamine B as tracer. The penetration of rhodamine B was uniform, deeper, and two times faster into epidermis than control gel. Conclusion: Conclusively, cavamax W7 composite ethosomes present themselves as efficient carrier for superior topical delivery of psoralen and have potential for clinical applications in minimizing side effects associated with photosensitivity of psoralen. PMID:24350036

  9. Site specificity of psoralen-DNA interstrand cross-linking determined by nuclease Bal31 digestion

    DEFF Research Database (Denmark)

    Zhen, W P; Buchardt, O; Nielsen, Henrik


    was analyzed, and these two psoralens showed identical site specificity. The 5'-TA preference is rationalized on the basis of the local DNA structure in terms of the pi-pi electronic interaction between the thymines and the intercalated psoralens, as well as on the base tilt angles of the DNA....

  10. Effects of nuclear isolation on psoralen affinity for chromatin

    Energy Technology Data Exchange (ETDEWEB)

    Inadomi, T.; Ross, P.M. (Rockefeller Univ., New York, NY (USA))


    We have tested the effects of nuclear isolation on intercalation of TMP (a psoralen) at specific sequences and in total DNA of cultured human cells. DNA in nuclei photobound about 20% more TMP than in cells and about 10% as much as purified DNA. In contrast, a transcribed ras gene and a randomly selected polymorphic sequence each bound about 20% more TMP than total DNA in cells. However, in nuclei, as in purified DNA, both sequences were just as sensitive as total DNA. Apparently, chromatin in cells exists within diverse TMP-binding environments and some of this diversity was lost upon nuclear isolation.

  11. Psoralen-containing sunscreen is tumorigenic in hairless mice

    Energy Technology Data Exchange (ETDEWEB)

    Cartwright, L.E.; Walter, J.F.


    Sunscreens containing 5-methoxypsoralen (5-MOP) are currently being marketed to promote tanning by inducing psoralen-mediated ultraviolet (UV) A (320-400 nm) melanogenesis. The rationale is that this may prevent UVB (290-320 nm) radiation-induced skin damage. However, mouse studies have shown that 5-MOP has the same cutaneous photocarcinogenic potential as 8-methoxypsoralen. In addition, the 5-MOP--containing sunscreen Sun System III (SS III), when combined with UVA, induces epidermal ornithine decarboxylase activity, an enzyme associated with tumor promotion. Therefore, we investigated whether SS III had sufficient psoralen concentration to be tumorigenic in hairless mice exposed to chronic, intermittent UVA radiation. SS III was applied to hairless mice 5 days per week for 20 weeks. After each application the mice were exposed to 2.5 to 10 joules/cm2 UVA radiation. All test groups developed atypical squamous papillomas in direct proportion to the dosage of UVA radiation received. A shorter latency period for tumor development was seen with larger UVA doses. Test animals followed up to 1 year developed invasive squamous cell tumors. Control groups (SS III without UVA and UVA without SS III) remained free of tumors. Animals receiving SS III plus UVA developed persistent skin thickening and increased dermal cyst formation similar to that reported with chronic exposure to UVB, a known carcinogenic wavelength.

  12. Evaluation of psoralen ethosomes for topical delivery in rats by using in vivo microdialysis. (United States)

    Zhang, Yong-Tai; Shen, Li-Na; Zhao, Ji-Hui; Feng, Nian-Ping


    This study aimed to improve skin permeation and deposition of psoralen by using ethosomes and to investigate real-time drug release in the deep skin in rats. We used a uniform design method to evaluate the effects of different ethosome formulations on entrapment efficiency and drug skin deposition. Using in vitro and in vivo methods, we investigated skin penetration and release from psoralen-loaded ethosomes in comparison with an ethanol tincture. In in vitro studies, the use of ethosomes was associated with a 6.56-fold greater skin deposition of psoralen than that achieved with the use of the tincture. In vivo skin microdialysis showed that the peak concentration and area under the curve of psoralen from ethosomes were approximately 3.37 and 2.34 times higher, respectively, than those of psoralen from the tincture. Moreover, it revealed that the percutaneous permeability of ethosomes was greater when applied to the abdomen than when applied to the chest or scapulas. Enhanced permeation and skin deposition of psoralen delivered by ethosomes may help reduce toxicity and improve the efficacy of long-term psoralen treatment.

  13. In vitro acetylcholinesterase inhibition by psoralen using molecular docking and enzymatic studies

    Directory of Open Access Journals (Sweden)

    Gauresh Somani


    Full Text Available Introduction: Alzheimer′s disease (AD has increased at an alarming rate and is now a worldwide health problem. Inhibitors of acetylcholinesterase (AChE leading to inhibition of acetylcholine breakdown constitute the main therapeutic strategy for AD. Psoralen was investigated as inhibitor of AChE enzyme in an attempt to explore its potential for the management of AD. Materials and Methods: Psoralen was isolated from powdered Psoralea corylifolia fruits. AChE enzyme inhibitory activity of different concentrations of psoralen was investigated by use of in vitro enzymatic and molecular docking studies. Further, the enzyme kinetics were studied using Lineweaver-Burk plot. Results: Psoralen was found to inhibit AChE enzyme activity in a concentration-dependent manner. Kinetic studies showed psoralen inhibits AChE in a competitive manner. Molecular docking study revealed that psoralen binds well within the binding site of the enzyme showing interactions such as π-π stacking and hydrogen bonding with residues present therein. Conclusion: The result of AChE enzyme inhibitory activity of the psoralen in this study is promising. It could be further explored as a potential candidate for further development of new drugs against AD.

  14. Psoralen loaded liposomal nanocarriers for improved skin penetration and efficacy of topical PUVA in psoriasis. (United States)

    Doppalapudi, Sindhu; Jain, Anjali; Chopra, Dhiraj Kumar; Khan, Wahid


    Psoralen in combination with ultraviolet A radiation (PUVA) is an FDA recommended therapy for clinical application in the management of severe recalcitrant psoriasis. Psoralen acts by intercalation of DNA and upon exposure to UV-A, it forms monoadducts which in turn induce apoptosis. Poor skin deposition, weak percutaneous permeability of psoralen and adverse effects of severe burning, blisters, pigmentation associated with conventional topical psoralen vehicles hinders the therapeutic efficacy and safety of topical PUVA. The aim of the present study is to formulate psoralen loaded liposomal nanocarriers for enhanced skin penetration, safety and efficacy of topical PUVA in psoriasis. Two different liposomal compositions i.e., cationic liposomes composed of DC-Chol, cholesterol and anionic liposomes composed of egg lecithin, cholesterol, tetramyristoyl cardiolipin were prepared for the topical delivery of psoralen. Liposomal carriers were characterized with respect to size, zeta potential, entrapment efficiency, stability, in vitro drug release and in vivo studies. Both liposomes were prepared with particle size of nearly 100nm. Zeta potential and entrapment efficiency of cationic liposomes were +25.8mV, 75.12% and anionic liposomes were -28.5mV, 60.08% respectively. Liposomal dermal distribution demonstrated higher penetration of both liposomal carriers over solution. Similarly, skin permeation study indicated 5 fold increase in permeation of psoralen with liposomal carriers. Topical application of psoralen liposomal gels on imiquimod induced psoriatic plaque model reduced the symptoms of psoriasis and levels of key psoriatic cytokines such as tumor necrosis factor-α, IL-17 and IL-22. In conclusion, the developed liposomal carriers of psoralen were found to be promising and can find application for optimal safety and efficacy of topical PUVA in psoriasis. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Induction of mixed-function oxidases in mouse liver by psoralens. (United States)

    Mandula, B B; Pathak, M A; Nakayama, Y; Davidson, S J


    8-methoxypsoralen, the most widely used psoralen in photochemotherapy, was shown to induce mixed-function oxidases. Induction of mixed-function oxidases in mouse liver was examined after mice were given a single dose of one of three psoralens. 8-methoxypsoralen clearly induced p-nitroanisole O-demethylase and slightly induced aryl hydrocarbon hydroxylase. Psoralen and 4,5',8-trimethylpsoralen failed to induce either enzyme. These results represent the first enzyme induction studies on these clinically useful compounds. The studies may have relevance in understanding the clinical differences in skin photosensitizing activity and photochemotherapeutic effectiveness of the compounds in such diseases as psoriasis and vitiligo.

  16. Sensitivity of Escherichia coli acrA Mutants to Psoralen plus Near-Ultraviolet Radiation

    DEFF Research Database (Denmark)

    Hansen, M. Trier


    The sensitivity to psoralen plus near-ultraviolet radiation (PUVA) was compared in a pair of E. coli strains differing at the acrA locus. Survival was determined for both bacteria and phage λ. AcrA mutant cells were 40 times more sensitive than wild type to the lethal effect of PUVA. Free λ phage...... of radiolabelled psoralens in the mutant. Binding was increased specifically to DNA rather than to nucleic acids in general. The difference in psoralen-binding ability determined by the acrA gene persisted after permeabilizing treatment of the cells. The results suggest that the acrA mutation causes an alteration...

  17. A method for fast determination of psoralens in oral solutions of phytomedicines using liquid chromatography. (United States)

    Pires, Adriana Elias; Honda, Neli Kiko; Cardoso, Cláudia Andréa Lima


    A method for sample preparation and analysis by high performance liquid chromatography with UV detection (HPLC-UV) has been developed for routine analysis of psoralen and bergapten, photosensitizing compounds, in oral solutions of phytomedicines employed in Brazil for some illnesses. The linearity, accuracy, the inter- and intra-day precision of the procedure were evaluated. Calibration curves for psoralen and bergapten were linear in the range of 1.0-600.0 microg ml(-1) and 1.0-400.0 microg ml(-1) respectively. The recoveries of the psoralens in the oral solutions analysed were 94.43-99.97%. The percentage coefficient of variation (CV) of the quantitative analysis of the psoralens in the products analysis was within 5%. In inter-equipment study was employed gas chromatography-flame ionization (CG-FID) detection.

  18. Isolation and characterization of pyrimidine-psoralen photoadducts from DNA

    Energy Technology Data Exchange (ETDEWEB)

    Straub, K.; Kanne, D.; Hearst, J.E.; Rapoport, H.


    We have examined the photoadducts of 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT) and native DNA. Five nucleoside-HMT monoaddition products have been isolated and characterized, corresponding to three deoxythymidine-HMT and two deoxyuridine (derived from deoxycytidine)-HMT adducts. Structural assignments are based on high resolution mass spectrometry and /sup 1/H NMR studies, including homonuclear spin decoupling and nuclear Overhauser effect (NOE) experiments. The results of this study indicate that (1) a limited number of nucleoside-psoralen adducts are formed with native, double-stranded DNA, and (2) the sterochemistry of the adducts is apparently determined by the geometry of the noncovalent intercalative complex formed by HMT and DNA prior to irradiation. 8 figures, 8 tables.

  19. Possible mechanism of psoralen phototoxicity not involving direct interaction with DNA

    Energy Technology Data Exchange (ETDEWEB)

    Laskin, J.D.; Lee, E.; Yurkow, E.J.; Laskin, D.L.; Gallo, M.A.


    Psoralens in combination with ultraviolet light (UVA; 320-400 nm) are used in the photochemical treatment of a variety of skin diseases including vitiligo, a skin depigmentational disorder, and psoriasis, a disease of accelerated epidermal cell proliferation. Although it is generally assumed that the major site of action of the psoralens is DNA, the authors have obtained evidence that another site may be the primary target for these compounds. They have identified specific, saturable, high-affinity binding sites for 8-methoxypsoralen on HeLa cells and have detected specific binding of 8-methoxypsoralen to four other human cell lines and five mouse cell lines. In HeLa cells, specific binding is reversible and independent of the ability of the compound to intercalate into DNA. In addition, binding sites become covalently modified by the psoralen after UVA exposure. Specific binding of 8-(methyoxy-/sup 3/H)methoxypsoralen constitutes 79% of the label bound to the cells. Scatchard analysis indicated two classes of psoralen binding sites. Based on these findings, the authors hypothesize that specific binding sites for psoralens on mammalian cells mediate, at least in part, psoralen-induced phototoxicity.

  20. Evaluation of skin viability effect on ethosome and liposome-mediated psoralen delivery via cell uptake. (United States)

    Zhang, Yong-Tai; Shen, Li-Na; Wu, Zhong-Hua; Zhao, Ji-Hui; Feng, Nian-Ping


    This study investigated the effect of skin viability on its permeability to psoralen delivered by ethosomes, as compared with liposomes. With decreasing skin viability, the amount of liposome-delivered psoralen that penetrated through the skin increased, whereas skin deposition of psoralen from both ethosomes and liposomes reduced. Psoralen delivery to human-immortalized epidermal cells was more effective using liposomes, whereas delivery to human embryonic skin fibroblast cells was more effective when ethosomes were used. These findings agreed with those of in vivo studies showing that skin psoralen deposition from ethosomes and liposomes first increased and then plateaued overtime, which may indicate gradual saturation of intracellular drug delivery. It also suggested that the reduced deposition of ethosome- or liposome-delivered psoralen in skin with reduced viability may relate to reduced cellular uptake. This work indicated that the effects of skin viability should be taken into account when evaluating nanocarrier-mediated drug skin permeation. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

  1. Psoralen-deoxyribonucleic acid photoreaction. Characterization of the monoaddition products from 8-methoxypsoralen and 4,5',8-trimethylpsoralen

    Energy Technology Data Exchange (ETDEWEB)

    Kanne, D.; Straub, K.; Rapoport, H.; Hearst, J.E.


    The isolation and structural characterization are described of the major monoaddition products formed in the photoreaction of two naturally occurring psoralens, 8-methoxypsoralen and 4,5',8-trimethylpsoralen, with high molecular weight, double-stranded DNA. Hydrolysis of the psoralen-modified DNA and subsequent chromatography resulted in the isolation of four modified nucleosides from each psoralen. Structural characterization was accomplished by mass spectrometry and /sup 1/H NMR analysis. The major products, accounting for 44 to 52% of the covalently bound psoralen, are two diastereomeric thymidine adducts formed by cycloaddition between the 5,6 double bond of the pyrimidine and the 4',5' (furan) double bond of the psoralen. All of the isolated adducts have cis-syn stereochemistry. The stereochemistry and product distribution of the adducts are determined in part by the constraints imposed by the DNA helix on the geometry of the noncovalent intercalation complex formed by psoralen and DNA prior to irradiation.

  2. The application of psoralens to the study of DNA structure, function and dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Spielmann, Peter Hans [Univ. of California, Berkeley, CA (United States). Dept. of Chemistry


    A series of six nitroxide spin-labeled psoralens were designed, synthesized and tested as probes for DNA dynamics. The synthesis of these spin-labeled psoralen derivatives and their photoreactivity with double-stranded DNA fragments is described. The spin labels (nitroxides) were demonstrated to survive the uv irradiation required to bind the probe to the target DNA. EPR spectra of the photobound spin-labels indicate that they do not wobble with respect to the DNA on the time-scales investigated. The author has used psoralen modified DNA as a model for the study of DNA repair enzyme systems in human cell free extracts. He has shown that damage-induced DNA synthesis is associated with removal of psoralen adducts and therefore is "repair synthesis" and not an aberrant DNA synthesis reaction potentiated by deformation of the DNA by adducts. He has found that all DNA synthesis induced by psoralen monoadducts is the consequence of removal of these adducts. By the same approach he has obtained evidence that this in vitro system is capable of removing psoralen cross-links as well. Reported here are synthetic methods that make use of high intensity lasers coupled with HPLC purification to make homogeneous and very pure micromole quantities of furan-side monoadducted, cross-linked, and pyrone-side monoadducted DNA oligonucleotide. These molecules are currently being studied by NMR and X-ray crystallography. The application of the site-specifically psoralen modified oligonucleotide synthesized by these methods to the construction of substrates for the investigation of DNA repair is also discussed.

  3. Comparison of ethosomes and liposomes for skin delivery of psoralen for psoriasis therapy. (United States)

    Zhang, Yong-Tai; Shen, Li-Na; Wu, Zhong-Hua; Zhao, Ji-Hui; Feng, Nian-Ping


    Recent reports have indicated that psoriasis may be caused by malfunctioning dermal immune cells, and psoralen ultraviolet A (PUVA) is an effective treatment for this chronic disease. However, conventional topical formulations achieve poor drug delivery across patches of psoriasis to their target sites. The present study describes the development of a novel psoralen transdermal delivery system employing ethosomes, flexible vesicles that can penetrate the stratum corneum and target deep skin layers. An in vitro skin permeation study showed that the permeability of psoralen-loaded ethosomes was superior to that of liposomes. Using ethosomes, psoralen transdermal flux and skin deposition were 38.89±0.32 μg/cm(2)/h and 3.87±1.74 μg/cm(2), respectively, 3.50 and 2.15 times those achieved using liposomes, respectively. The ethosomes and liposomes were found to be safe following daily application to rat skin in vivo, for 7 days. The ethosomes showed better biocompatibility with human embryonic skin fibroblasts than did an equivalent ethanol solution, indicating that the phosphatidylcholine present in ethosome vesicles improved their biocompatibility. These findings indicated that ethosomes could potentially improve the dermal and transdermal delivery of psoralen and possibly of other drugs requiring deep skin delivery. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Probing the binding mode of psoralen to calf thymus DNA. (United States)

    Zhou, Xiaoyue; Zhang, Guowen; Wang, Langhong


    The binding properties between psoralen (PSO) and calf thymus DNA (ctDNA) were predicted by molecular docking, and then determined with the use of UV-vis absorption, fluorescence, circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy, coupled with DNA melting and viscosity measurements. The data matrix obtained from UV-vis spectra was resolved by multivariate curve resolution-alternating least squares (MCR-ALS) approach. The pure spectra and the equilibrium concentration profiles for PSO, ctDNA and PSO-ctDNA complex extracted from the highly overlapping composite response were obtained simultaneously to evaluate the PSO-ctDNA interaction. The intercalation mode of PSO binding to ctDNA was supported by the results from the melting studies, viscosity measurements, iodide quenching and fluorescence polarization experiments, competitive binding investigations and CD analysis. The molecular docking prediction showed that the specific binding most likely occurred between PSO and adenine bases of ctDNA. FT-IR spectra studies further confirmed that PSO preferentially bound to adenine bases, and this binding decreased right-handed helicity of ctDNA and enhanced the degree of base stacking with the preservation of native B-conformation. The calculated thermodynamic parameters indicated that hydrogen bonds and van der Waals forces played a major role in the binding process. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Psoralen Inactivation of Viruses: A Process for the Safe Manipulation of Viral Antigen and Nucleic Acid

    Directory of Open Access Journals (Sweden)

    Katherine Schneider


    Full Text Available High consequence human pathogenic viruses must be handled at biosafety level 2, 3 or 4 and must be rendered non-infectious before they can be utilized for molecular or immunological applications at lower biosafety levels. Here we evaluate psoralen-inactivated Arena-, Bunya-, Corona-, Filo-, Flavi- and Orthomyxoviruses for their suitability as antigen in immunological processes and as template for reverse transcription PCR and sequencing. The method of virus inactivation using a psoralen molecule appears to have broad applicability to RNA viruses and to leave both the particle and RNA of the treated virus intact, while rendering the virus non-infectious.

  6. Targeted gene correction using psoralen, chlorambucil and camptothecin conjugates of triplex forming peptide nucleic acid (PNA)

    DEFF Research Database (Denmark)

    Birkedal, Henrik; Nielsen, Peter E


    Gene correction activation effects of a small series of triplex forming peptide nucleic acid (PNA) covalently conjugated to the DNA interacting ligands psoralen, chlorambucil and camptothecin targeted proximal to a stop codon mutation in an EGFP reporter gene were studied. A 15-mer homopyrimidine...

  7. Isolation and purification of psoralen and isopsoralen and their efficacy and safety in the treatment of osteosarcoma in nude rats. (United States)

    Lu, Honghui; Zhang, Lihai; Liu, Daohong; Tang, Peifu; Song, Feixiang


    Modern studies have shown that psoralen has a significant inhibitory effect on tumor growth in a variety of animals and humans. To obtain coumarin compounds - psoralen and isopsoralen - from traditional Chinese medicine Psoralea corylifolia L. using chromatographic techniques and isolation and purification methods, and to observe the transplanted tumor growth inhibitory effects and adverse reactions of psoralen and isopsoralen in nude rats with osteosarcoma. Dried ripe fruits of Psoralea corylifolia L. were taken as the raw material to prepare crude extract of Psoralea corylifolia L. by ethanol reflux method. Column chromatography was used to isolate the crude extract; compounds were structurally identified based on (1)H-NMR, (13)C-NMR spectra, the two compounds were identified as psoralen andisopsoralen, and their contents were 99.7% and 99.6, respectively. Nude rat model of osteosarcoma was established; the rats were randomized into: normal saline group, psoralen low- and high-dose groups, isopsoralen low- and high-dose groups, and cisplatin group. Osteosarcoma volume and weight inhibition rates in nude rats in each group were observed; radioimmunoassay was used to determine the serum alkaline phosphatase activity; peripheral blood cell and bone marrow nucleated cell counts were determined; light microscopy was used to observe heart, liver, spleen, lung, kidney, and tumor histopathology; and electron microscopy was used to observe the fine structure of tumor cells. Tumor volume inhibition rates were 43.75% and 40.18%, respectively, in the psoralen and isopsoralen low-dose groups, and tumor weight inhibition rates were 38.83% and 37.77%. Tumor volume inhibition rates were 67.86% and 66.96%, respectively, in the psoralen and isopsoralen high-dose groups, and tumor weight inhibition rates were 49.47% and 47.87%. Psoralen and ispsoralen markedly lowered serum AKP level. Psoralen and isopsoralen induced apoptosis or necrosis of osteosarcoma. After administration of

  8. Mechanisms Underlying the Antiproliferative and Prodifferentiative Effects of Psoralen on Adult Neural Stem Cells via DNA Microarray

    Directory of Open Access Journals (Sweden)

    You Ning


    Full Text Available Adult neural stem cells (NSCs persist throughout life to replace mature cells that are lost during turnover, disease, or injury. The investigation of NSC creates novel treatments for central nervous system (CNS injuries and neurodegenerative disorders. The plasticity and reparative potential of NSC are regulated by different factors, which are critical for neurological regenerative medicine research. We investigated the effects of Psoralen, which is the mature fruit of Psoralea corylifolia L., on NSC behaviors and the underlying mechanisms. The self-renewal and proliferation of NSC were examined. We detected neuron- and/or astrocyte-specific markers using immunofluorescence and Western blotting, which could evaluate NSC differentiation. Psoralen treatment significantly inhibited neurosphere formation in a dose-dependent manner. Psoralen treatment increased the expression of the astrocyte-specific marker but decreased neuron-specific marker expression. These results suggested that Psoralen was a differentiation inducer in astrocyte. Differential gene expression following Psoralen treatment was screened using DNA microarray and confirmed by quantitative real-time PCR. Our microarray study demonstrated that Psoralen could effectively regulate the specific gene expression profile of NSC. The genes involved in the classification of cellular differentiation, proliferation, and metabolism, the transcription factors belonging to Ets family, and the hedgehog pathway may be closely related to the regulation.

  9. Development and in vitro assessment of psoralen and resveratrol co-loaded ultradeformable liposomes for the treatment of vitiligo. (United States)

    Doppalapudi, Sindhu; Mahira, Shaheen; Khan, Wahid


    Vitiligo is a de-pigmenting skin disorder characterized by white patches on skin due to partial or complete loss of melanocytes. Psoralen in combination with ultraviolet-A (PUVA) acts by stimulation of melanin content and tyrosinase activity in melanocytes. Resveratrol, a sirtuin activator and a potential anti-oxidant reduce oxidative stress which is one of the triggering factors for initiation of vitiligo. Despite their therapeutic activity, weak percutaneous permeability of psoralen and poor solubility of resveratrol hinders their effective topical administration. The aim of present study is to formulate ultradeformable liposomes (UDL) co-loaded with psoralen and resveratrol for evaluation of PUVA and anti-oxidant combination in vitiligo treatment. For this purpose, UDL composed of DC-Chol, cholesterol and sodium deoxy cholate were prepared for their co-delivery. Liposomal carriers were characterized and evaluated for their efficacy using B16F10 cell line. Free radical scavenging potential was also determined for these carriers by in vitro anti-oxidant assays. Optimal co-loaded UDL with particle size ranging from 120 to 130nm, zeta potential of +46.2mV, entrapment efficiency of 74.09% (psoralen) and 76.91% (resveratrol) were obtained. Compared to control, co-loaded UDL showed significant stimulation of melanin and tyrosinase activity with major contribution of psoralen. Further, co-loaded UDL also exhibited potential free radical scavenging activity where resveratrol played a key role. Hence, psoralen and resveratrol co-loaded UDL acts in vitiligo through dual mechanisms of action viz., stimulation of melanin and tyrosinase activity as well as by anti-oxidant activity. These findings indicate that psoralen and resveratrol co-loaded UDL has the promising therapeutic potential for the treatment of vitiligo. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Phototoxic effects of Hypericum extract in cultures of human keratinocytes compared with those of psoralen. (United States)

    Bernd, A; Simon, S; Ramirez Bosca, A; Kippenberger, S; Diaz Alperi, J; Miquel, J; Villalba Garcia, J F; Pamies Mira, D; Kaufmann, R


    Extracts of Hypericum perforatum (St. John's wort) are used in the treatment of depression. They contain the plant pigment hypericin and hypericin derivates. These compounds have light-dependent activities. In order to estimate the potential risk of phototoxic skin damage during antidepressive therapy, we investigated the phototoxic activity of hypericin extract using cultures of human keratinocytes and compared it with the effect of the well-known phototoxic agent psoralen. The absorbance spectrum of our Hypericum extract revealed maxima in the whole UV range and in parts of the visible range. We cultivated human keratinocytes in the presence of different Hypericum concentrations and irradiated the cells with 150 mJ/cm2 UVB, 1 J/cm2 UVA or 3 h with a white light of photon flux density 2.6 mumol m-2 s-1. The determination of the bromodeoxyuridine incorporation rate showed a concentration- and light-dependent decrease in DNA synthesis with high hypericin concentrations (> or = 50 micrograms/mL) combined with UVA or visible light radiation. In the case of UVB irradiation a clear phototoxic cell reaction was not detected. We found phototoxic effects even with 10 ng/mL psoralen using UVA with the same study design as in the case of the Hypericum extract. These results confirm the phototoxic activity of Hypericum extract on human keratinocytes. However, the blood levels that are to be expected during antidepressive therapy are presumably too low to induce phototoxic skin reactions.

  11. Orally administered Polypodium leucotomos extract decreases psoralen-UVA-induced phototoxicity, pigmentation, and damage of human skin

    NARCIS (Netherlands)

    Middelkamp-Hup, Maritza A.; Pathak, Madhu A.; Parrado, Concepcion; Garcia-Caballero, Tomas; Rius-Díaz, Francisca; Fitzpatrick, Thomas B.; González, Salvador


    BACKGROUND: The use of psoralen-UVA (PUVA) in patients of skin phototype I to II is limited by side effects of acute phototoxicity and possible long-term carcinogenesis. OBJECTIVE: We sought to assess oral Polypodium leucotomos (PL) extract in decreasing PUVA-induced phototoxicity of human skin on a

  12. Using Limes and Synthetic Psoralens to Enhance Solar Disinfection of Water (SODIS): A Laboratory Evaluation with Norovirus, Escherichia coli, and MS2


    Harding, Alexander S.; Schwab, Kellogg J.


    We investigated the use of psoralens and limes to enhance solar disinfection of water (SODIS) using an UV lamp and natural sunlight experiments. SODIS conditions were replicated using sunlight, 2 L polyethylene terephthalate (PET) bottles, and tap water with Escherichia coli, MS2 bacteriophage, and murine norovirus (MNV). Psoralens and lime acidity both interact synergistically with UV radiation to accelerate inactivation of microbes. Escherichia coli was ablated > 6.1 logs by SODIS + Lime Sl...

  13. Rescue of Mitomycin C- or Psoralen-Inactivated Micrococcus Radiodurans by Additional Exposure to Radiation or Alkylating Agents

    DEFF Research Database (Denmark)

    Hansen, M. Trier


    The processing of damaged DNA was altered in a mitomycin C-sensitive mutant (mtcA) of Micrococcus radiodurans. Even though the mutant retained resistance to 254-nm UV radiation, it did not, in contrast to the wild-type strain, show any excessive DNA degradation or cell death when incubated...... in the mtcA background. In this strain, additional damage infficted upon the cellular DNA effected a massive rescue of cells previously inactivated by mitomycin C. Rescue was provoked by ionizing radiation, by UV light, or by simple alkylating agents. Cells treated with psoralen plus near-UV radiation could...... interstrand cross-links produced by mitomycin C or psoralen plus near-UV light, but induced only by the more abundant number of damages produced by radiation or simple alkylating agents....

  14. Psoralen plus ultraviolet A (PUVA soaks and UVB TL01 treatment for chronic hand dermatoses

    Directory of Open Access Journals (Sweden)

    Lisbeth Jensen


    Full Text Available Chronic eczematous hand dermatoses with and without contact allergies are complex diseases, which makes it a challenge to select the best treatment and obtain an optimal patient experience and a satisfactory treatment result. The aim of this study was to evaluate retrospectively the clinical effect and patient experience of local treatment with psoralen plus ultraviolet A (PUVA soaks and TL01 phototherapy for severe chronic hand dermatoses, and also to evaluate the quality of life for the subgroup of patients with allergic contact dermatitis including Compositae allergy. A retrospective evaluation of results for 94 consecutive patients having received a total of 121 treatment courses with local PUVA soaks or TL01 phototherapy for one of the following diagnoses (n=number of treatment courses: psoriasis (n=19, hyperkeratotic hand eczema (n=27, Pustulosis Palmoplantaris (PPP (n=22, vesicular eczema (n=16, Compositae dermatitis (n=24, and allergic contact dermatitis (n=13. Moreover, semi-structured interviews with 6 selected patients having multiple contact allergies including Compositae allergy were used to evaluate quality of life. As a result, we found that PUVA soaks has good effect in patients with psoriasis and hyperkeratotic hand eczema and local phototherapy for chronic hand dermatoses is a useful treatment option in selected cases.

  15. Photo-crosslinked hyaluronic acid coated upconverting nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Mrazek, Jiri, E-mail:; Kettou, Sofiane; Matuska, Vit; Svozil, Vit; Huerta-Angeles, Gloria; Pospisilova, Martina; Nesporova, Kristina; Velebny, Vladimir [Contipro a. s. (Czech Republic)


    Hyaluronic acid (HA)-coated inorganic nanoparticles display enhanced interaction with the CD44 receptors which are overexpressed in many types of cancer cells. Here, we describe a modification of core-shell β-NaY{sub 0.80}Yb{sub 0.18}Er{sub 0.02}F{sub 4}@NaYF{sub 4} nanoparticles (UCNP) by HA derivative bearing photo-reactive groups. UCNP capped with oleic acid were firstly transferred to aqueous phase by an improved protocol using hydrochloric acid or lactic acid treatment. Subsequently, HA bearing furanacryloyl moieties (HA-FU) was adsorbed on the nanoparticle surface and crosslinked by UV irradiation. The crosslinking resulted in stable HA coating, and no polymer desorption was observed. As-prepared UCNP@HA-FU show a hydrodynamic diameter of about 180 nm and are colloidally stable in water and cell culture media. The cellular uptake by normal human fibroblasts and MDA MB-231 cancer cell line was investigated by upconversion luminescence imaging.

  16. Photo-crosslinked hyaluronic acid coated upconverting nanoparticles (United States)

    Mrazek, Jiri; Kettou, Sofiane; Matuska, Vit; Svozil, Vit; Huerta-Angeles, Gloria; Pospisilova, Martina; Nesporova, Kristina; Velebny, Vladimir


    Hyaluronic acid (HA)-coated inorganic nanoparticles display enhanced interaction with the CD44 receptors which are overexpressed in many types of cancer cells. Here, we describe a modification of core-shell β-NaY0.80Yb0.18Er0.02F4@NaYF4 nanoparticles (UCNP) by HA derivative bearing photo-reactive groups. UCNP capped with oleic acid were firstly transferred to aqueous phase by an improved protocol using hydrochloric acid or lactic acid treatment. Subsequently, HA bearing furanacryloyl moieties (HA-FU) was adsorbed on the nanoparticle surface and crosslinked by UV irradiation. The crosslinking resulted in stable HA coating, and no polymer desorption was observed. As-prepared UCNP@HA-FU show a hydrodynamic diameter of about 180 nm and are colloidally stable in water and cell culture media. The cellular uptake by normal human fibroblasts and MDA MB-231 cancer cell line was investigated by upconversion luminescence imaging.

  17. The direct observation of a psoralen-thymine UVA induced solid-state cycloaddition reaction product by single-crystal x-ray diffractometry. (United States)

    Pfluger, C E; Ostrander, R L


    Single-crystal x-ray diffraction methods have been used to directly observe and simultaneously determine the molecular structure of the UVA induced cis-syn photocycloaddition product in a partially photolyzed single crystal of a psoralen(pyrone ring side)-DNA(thymine) interaction model compound, 1'-(8-oxypsoralen)-8'(thym-1"yl)3',6'-dioxaoctane.

  18. Psoralen-mediated virus photoinactivation in platelet concentrates: enhanced specificity of virus kill in the absence of shorter UVA wavelengths

    Energy Technology Data Exchange (ETDEWEB)

    Margolis-Nunno, Henrietta; Robinson, Richard; Horowitz, Bernard; Ben-Hur, Ehud [New York Blood Center, NY (United States); Geacintov, N.E. [New York Univ., NY (United States). Dept. of Chemistry


    Treatments with psoralens and long-wavelength ultraviolet radiation (UVA, 320-400 nm; PUVA) have shown efficacy for virus sterilization of platelet concentrates (PC). We have employed the psoralen derivative 4`-aminomethyl-4,5`,8-trimethylpsoralen (AMT), and have found that platelet integrity is best preserved when rutin, a flavonoid that quenches multiple reactive oxygen species, is present during AMT/UVA treatment of PC. In this report, we examine the effects of different UVA spectra under our standard PC treatment conditions (i.e. 50 {mu}g/mL AMT, 0.35 mM rutin and 38 J/cm{sup 2} UVA). Added vesicular stomatitis virus (VSV; {>=} 5.5 log{sub 10}) was completely inactivated with the simultaneous maintenance of the platelet aggregation response (> 90% of control) when a UVA light source with transmission mainly between 360 and 370 nm (narrow UVA1) was used. In contrast, with a broad-band UVA (320-400 nm; broad UVA) light source, the aggregation response was greatly compromised (< 50% of control) with only a minor increase in the rate of VSV kill. With this lamp, platelet function could be improved to about 75% of the control by adding a long-pass filter, which reduced the transmission of shorter ({<=} 345 nm) UVA wavelengths (340-400 nm; UVA1). At equivalent levels of virus kill, aggregation function was always best preserved when narrow UVA1 was used for PUVA treatment. Even in the absence of AMT, and with or without rutin present, narrow UVA1 irradiation was better tolerated by platelets than was broad UVA. (author).

  19. Using limes and synthetic psoralens to enhance solar disinfection of water (SODIS): a laboratory evaluation with norovirus, Escherichia coli, and MS2. (United States)

    Harding, Alexander S; Schwab, Kellogg J


    We investigated the use of psoralens and limes to enhance solar disinfection of water (SODIS) using an UV lamp and natural sunlight experiments. SODIS conditions were replicated using sunlight, 2 L polyethylene terephthalate (PET) bottles, and tap water with Escherichia coli, MS2 bacteriophage, and murine norovirus (MNV). Psoralens and lime acidity both interact synergistically with UV radiation to accelerate inactivation of microbes. Escherichia coli was ablated > 6.1 logs by SODIS + Lime Slurry and 5.6 logs by SODIS + Lime Juice in 30-minute solar exposures, compared with a 1.5 log reduction with SODIS alone (N = 3; P 3.9 logs by SODIS + Lime Slurry, 1.9 logs by SODIS + Lime Juice, and 1.4 logs by SODIS in 2.5-hour solar exposures (N = 3; P SODIS, with SODIS against human norovirus should be investigated further.

  20. Allelism of Saccharomyces cerevisiae gene PSO10, involved in error-prone repair of psoralen-induced DNA damage, with SUMO ligase-encoding MMS21. (United States)

    Hoch, Nícolas C; Santos, Rafael S; Rosa, Renato M; Machado, Roseane M; Saffi, Jenifer; Brendel, Martin; Henriques, João A P


    In order to extend the understanding of the genetical and biochemical basis of photo-activated psoralen-induced DNA repair in the yeast Saccharomyces cerevisiae we have identified and cloned 10 pso mutants. Here, we describe the phenotypic characterization and molecular cloning of the pso10-1 mutant which is highly sensitive to photoactivated psoralens, UV(254) (nm) radiation and the alkylating agent methylmethane sulphonate. The pso10-1 mutant allele also confers a block in the mutagenic response to photoactivated psoralens and UV(254) (nm) radiation, and homoallelic diploids do not sporulate. Molecular cloning using a yeast genomic library, sequence analysis and genetic complementation experiments proved pso10-1 to be a mutant allele of gene MMS21 that encodes a SUMO ligase involved in the sumoylation of several DNA repair proteins. The ORF of pso10-1 contains a single nucleotide C-->T transition at position 758, which leads to a change in amino acid sequence from serine to phenylalanine [S253F]. Pso10-1p defines a leaky mutant phenotype of the essential MMS21 gene, and as member of the Smc5-Smc6 complex, still has some essential functions that allow survival of the mutant. DNA repair via translesion synthesis is severely impaired as the pso10-1 mutant allele confers severely blocked induced forward and reverse mutagenesis and shows epistatic interaction with a rev3Delta mutant allele. By identifying the allelism of PSO10 and MMS21 we demonstrate the need of a fully functional Smc5-Smc6 complex for a WT-like adequate repair of photoactivated psoralen-induced DNA damage in yeast.

  1. SU-E-T-93: Activation of Psoralen at Depth Using Kilovoltage X-Rays: Physics Considerations in Implementing a New Teletherapy Paradigm

    Energy Technology Data Exchange (ETDEWEB)

    Adamson, J; Yoon, P; Liu, L; Alcorta, D; Spector, N; Oldham, M [Duke University Medical Center, Durham, NC (United States); Nolan, M; Gieger, T [North Carolina State University, Raleigh, NC (United States); Walder, H; Fathi, Z; Beyer, W [Immunolight LLC, Detroit, MI (United States)


    Purpose: Psoralen is a UV-light activated anti-cancer biotherapeutic used for treating skin lesions (PUVA) and advanced cutaneous T-cell lymphoma (ECP). To date psoralen has not been used to treat deep seated tumors due to difficulty in generating UV-light at depth. We recently demonstrated psoralen activation at depth by introducing energy converting particles that absorb kV x-ray radiation and re-emit UV-light. Our in-vitro work found that 0.2–1Gy using 40–100kVp x-rays combined with psoralen and particles can induce a substantial apoptotic response beyond that expected from the sum of individual components. In preparation for a phase I clinical trial of canine companion animals, we address the physics and dosimetry considerations for applying this new teletherapy paradigm to an in-vivo setting. Methods: The kV on-board imaging (OBI) system mounted on a medical linear accelerator (Varian) was commissioned to deliver the prescribed dose (0.6Gy) using 80 and 100kVp. Dosimetric measurements included kVp, HVL, depth dose, backscatter factors, collimator and phantom scatter factors, field size factors, and blade leakage. Absolute dosimetry was performed following AAPM TG61 recommendations and verified with an independent kV dose meter. We also investigated collimated rotational delivery to minimize skin dose using simple dose calculations on homogeneous cylindrical phantoms. Results: Single beam delivery is feasible for shallow targets (<5cm) without exceeding skin tolerance, while a rotational delivery may be utilized for deeper targets; skin dose is ∼75% of target dose for 80kVp collimated rotational delivery to a 3cm target within a 20cm phantom. Heat loading was tolerable; 0.6Gy to 5cm can be delivered before the anode reaches 75% capacity. Conclusion: KV teletherapy for Psoralen activation in deep seated tissue was successfully commissioned for a Varian OBI machine for use in a phase I clinical trial in canines. Future work will use Monte Carlo dosimetry to

  2. The RecQ helicase RECQL5 participates in psoralen-induced interstrand cross-link repair (United States)

    Bohr, Vilhelm A.


    Interstrand cross-links (ICLs) are very severe lesions as they are absolute blocks of replication and transcription. This property of interstrand cross-linking agents has been exploited clinically for the treatment of cancers and other diseases. ICLs are repaired in human cells by specialized DNA repair pathways including components of the nucleotide excision repair pathway, double-strand break repair pathway and the Fanconi anemia pathway. In this report, we identify the role of RECQL5, a member of the RecQ family of helicases, in the repair of ICLs. Using laser-directed confocal microscopy, we demonstrate that RECQL5 is recruited to ICLs formed by trioxalen (a psoralen-derived compound) and ultraviolet irradiation A. Using single-cell gel electrophoresis and proliferation assays, we identify the role of RECQL5 in the repair of ICL lesions. The domain of RECQL5 that recruits to the site of ICL was mapped to the KIX region between amino acids 500 and 650. Inhibition of transcription and of topoisomerases did not affect recruitment, which was inhibited by DNA-intercalating agents, suggesting that the DNA structure itself may be responsible for the recruitment of RECQL5 to the sites of ICLs. PMID:23715498

  3. Increased prevalence of human papillomavirus in hairs plucked from patients with psoriasis treated with psoralen-UV-A. (United States)

    Wolf, Peter; Seidl, Hannes; Bäck, Barbara; Binder, Barbara; Höfler, Gerald; Quehenberger, Franz; Hoffmann, Christine; Kerl, Helmut; Stark, Sabine; Pfister, Herbert J; Fuchs, Pawel G


    Patients with psoriasis treated with psoralen-UV-A (PUVA) are at increased risk of skin cancer; however, the exact causes of this increased incidence are not well understood. It has been suggested that PUVA may increase expression of the tumorigenic agent human papillomavirus (HPV) in skin by directly stimulating virus replication, immune suppression, or both, thereby leading to skin cancer formation. To determine whether HPV DNA prevalence in the skin is increased after long-term PUVA treatment. Screening for the presence of HPV sequences in DNA isolated from plucked body hairs of patients with psoriasis with a history of PUVA exposure and a history of skin cancer (group A), PUVA exposure and no history of skin cancer (group B), and no PUVA exposure and no history of skin cancer (group C). University hospital. Hair samples were obtained from 81 patients with psoriasis (56 men and 25 women; mean age, 52 years), including 16 in group A (mean number of PUVA exposures, 702), 35 in group B (mean number of PUVA exposures, 282), and 30 in group C. DNA was isolated from the hair samples and analyzed by polymerase chain reaction with the use of 2 nested primer systems specific for epidermodysplasia verruciformis-associated or related and genital or mucosal virus types, respectively. The rate of HPV DNA positivity was significantly higher in groups A (73% [11/15]) and B (69% [24/35]) than in group C (36% [10/28]) (A + B vs C, P =.009; chi(2) test; age adjusted). Conclusion The prevalence of HPV in the skin (hair follicles) is increased in patients with psoriasis who have a history of PUVA exposure.

  4. Mutant pso8-1 of Saccharomyces cerevisiae, sensitive to photoactivated psoralens, UV radiation, and chemical mutagens, contains a rad6 missense mutant allele. (United States)

    Rolla, H; Grey, M; Schmidt, C L; Niegemann, E; Brendel, M; Henriques, J A P


    A novel mutant isolate of Saccharomyces cerevisiae, sensitive to photoactivated mono- and bi-functional psoralens, to UV at 254 nm (UVC), and to nitrosoguanidine, was found to complement the photoactivated psoralen-sensitivity phenotype conferred by the pso1- pso7 mutations and was therefore named pso8-1. A constructed pso8-1 rad4-4 double mutant was super-sensitive to UVC, thus indicating a synergistic interaction of the two mutant alleles. Molecular cloning via complementation of the pso8 mutant's sensitivity phenotype and genetic studies revealed that pso8 is allelic to RAD6. While a pso8-1 mutant had low mutagen-induced mutability, homoallelic diploids showed nearly wild-type sporulation. Sequence analysis of the mutant allele showed pso8-1 to contain a novel, hitherto undescribed T-->C transition in nucleotide position 191, leading to a substitution by leucine of a highly conserved proline at position 64, Rad6-[P64L], which may have severe consequences for the tertiary structure (and hence binding to Rad18p) of the mutant protein.

  5. Narrow-band ultraviolet B phototherapy versus broad-band ultraviolet B or psoralen-ultraviolet A photochemotherapy for psoriasis. (United States)

    Chen, Xiaomei; Yang, Ming; Cheng, Yan; Liu, Guan J; Zhang, Min


    The most commonly used types of phototherapy for treating psoriasis are narrow-band ultraviolet B (NB-UVB); broad-band ultraviolet B (BB-UVB), which includes selective (delivering radiation with a wavelength range of 305 to 325 nm) and conventional BB-UVB (280 to 320 nm); and psoralen ultraviolet A photochemotherapy (oral or bath PUVA). There is substantial controversy regarding their efficacy when compared with each other. To assess the effects of narrow-band ultraviolet B phototherapy versus broad-band ultraviolet B or psoralen ultraviolet A photochemotherapy for psoriasis. We searched the following databases up to August 2013: the Cochrane Skin Group Specialised Register, CENTRAL in The Cochrane Library (2013, Issue 7), MEDLINE (from 1946), and EMBASE (from 1974). We searched the following databases up to November 2012: CNKI (from 1974) and CBM (from 1978). We also searched trials registers and the OpenGrey database. We included all randomised controlled trials (RCTs) that compared NB-UVB phototherapy with BB-UVB or PUVA for treating psoriasis, which included chronic plaque psoriasis (CPP), guttate psoriasis (GP), and palmoplantar psoriasis (PPP). Two review authors independently conducted the study selection, 'Risk of bias' assessment, and data extraction. We included 13 RCTs, with a total of 662 participants. We report the results of intention-to-treat analyses (ITT) here. Our primary outcomes of interest were as follows: Participant-rated global improvement, Percentage of participants reaching Psoriasis Area and Severity Index (PASI) 75 (which meant equal to or more than 75% reduction in PASI score), Withdrawal due to side-effects, and Clearance rate.In one RCT of NB-UVB compared with oral PUVA in participants with CPP, the difference in PASI 75 was not statistically significant (risk ratio (RR) 0.91, 95% confidence interval (CI) 0.63 to 1.32; N = 51; low quality). In three other RCTs of CPP, the clearance rates were inconsistent because in one, there was no

  6. Psoriasis, Psoralen and Sunlight

    Directory of Open Access Journals (Sweden)

    R.P.C Naik


    Full Text Available Topical or oral administration of photoactive furocumarins followed by exposure to ultra violet light from artificial sources has been shown previously to clear psoriatic lesions. Sunlight has been chosen as the source of UVL m two separate paired comparison studies using topical and, oral 4,5, 8 trimethylpsoralen. Two out of 21 on topical therapy and none out of 6 patients on oral paired-comparison study showed faster clearance of the drug treated lesions compared to control sites.

  7. Molecular analysis by electron microscopy of the removal of psoralen-photoinduced DNA cross-links in normal and Fanconi's anemia fibroblasts

    Energy Technology Data Exchange (ETDEWEB)

    Rousset, S.; Nocentini, S.; Revet, B.; Moustacchi, E. (Institut Curie, Biologie, Paris (France))


    The induction and fate of psoralen-photoinduced DNA interstrand cross-links in the genome of Fanconi's anemia (FA) fibroblasts of complementation groups A and B, and of normal human fibroblasts, were investigated by quantitative analysis of totally denatured DNA fragments visualized by electron microscopy. 8-Methoxypsoralen (5 x 10(-5) M) interstrand cross-links were induced as a function of the near ultraviolet light dose. With time of postexposure incubation, a fraction of interstrand cross-links disappeared in all cell lines. However, 24 h after treatment, this removal was significantly lower in the two FA group A cell lines examined (34-39%) than in the FA group B and normal cell lines (43-53 and 47-57%, respectively). These data indicate that FA cells are at least able to recognize and incise interstrand cross-links, as normal cells do, although group A cells seem somewhat hampered in this process. This is in accord with data obtained on the same cell lines using another biochemical assay. Since the fate of cross-links in FA constituted a controversial matter, it is important to stress that two different methodologies applied to genetically well defined cell lines led to the same conclusions.

  8. Inactivation of viruses in platelet suspensions that retain their in vitro characteristics: Comparison of psoralen-ultraviolet A and merocyanine 540-visible light methods

    Energy Technology Data Exchange (ETDEWEB)

    Dodd, R.Y.; Moroff, G.; Wagner, S.; Dabay, M.H.; Dorfman, E.; George, V.; Ribeiro, A.; Shumaker, J.; Benade, L.E. (Jerome H. Holland Laboratory, American Red Cross, Rockville, MD (USA))


    The ability of two fundamentally different photochemical procedures to inactivate model viruses in platelet suspensions was compared. Merocyanine 540 (MC 540) with visible light was used as an example of an oxygen-dependent chemical-directed at the viral membrane, and aminomethyl trimethyl psoralen (AMT) with ultraviolet A light (UVA) was used as an example of a nucleic acid-directed system. Antiviral conditions in petri dishes were identified and the effects of these procedures on platelet suspensions in plastic storage containers were studied. Concentrations of photochemicals in the 10 to 150 mumol range with 30 to 60 minutes of visible light (MC 540) or 1 to 2 minutes of UVA (AMT) readily inactivated 5 to 6 log10 of vesicular stomatitis virus (VSV) and other model viruses in platelet suspensions, provided the plasma concentration was reduced to about 15 percent by the use of a synthetic platelet storage medium. Extracellular pH, morphology scores, and aggregation response dropped markedly when platelets were treated with MC 540 and visible light. However, treatment with 136 mumol per L of AMT and 1 to 3 minutes of UVA could inactivate 5 log10 of VSV in platelet suspensions with retention of platelet characteristics for 4 days, particularly if oxygen levels were reduced during treatment. These studies demonstrate that AMT-UVA treatment meets the initial requirements for virus inactivation in platelet suspensions.

  9. Injectable photo crosslinked enhanced double-network hydrogels from modified sodium alginate and gelatin. (United States)

    Yuan, Liu; Wu, Yu; Gu, Qi-Sheng; El-Hamshary, Hany; El-Newehy, Mohamed; Mo, Xiumei


    Recently, photocrosslinked hydrogels have attracted more and more attention in biomedical applications. In this study, a serials of injectable hydrogels were fabricated from aldehyde methacrylate sodium alginate and amino gelatin (AMSA/AG) using a two-step process. Here, sodium alginate, a kind of natural polysaccharide, was modified by oxidizer to form aldehyde sodium alginate (ASA), and methacrylate groups were further grafted on the main chain of ASA. Gelatin, the denatured form of collagen, was modified with ethylenediamine (ED) to graft more amino groups. When AMSA and AG aqueous solutions were mixed, the Schiff base reaction occurred quickly to form the primary network between aldehyde groups in AMSA and amino groups in AG, and then a 365nm ultraviolet (UV) light was used to initiate the radical reaction of methacrylate groups in AMSA to produce the secondary network. The structures and properties of AMSA/AG hydrogels were evaluated by Fourier Transforms Infrared spectroscopy (FTIR) and 1HNMR analysis. The swelling ratio confirmed the density of crosslinked networks, and the mechanical performance demonstrated that the UV initiated the double crosslinking network hydrogels have an improved mechanical properties compared to the single Schiff base networks hydrogels. The results showed that the photocrosslinked double network hydrogels have enhanced mechanical properties, good biocompatibility and controllable degradation rate. So, this hydrogels may have great potential utilized in regenerative medicine as therapeutic materials. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Photo-crosslinking, bio-inspired terpolymer adhesives intended for medical applications (United States)

    Harper, Tristan

    A bio-inspired, modular terpolymer adhesive has been synthesized containing three different functionalities: a photocrosslinking segment, wet adhesion segment, and a water soluble segment. Wet adhesion is brought on by an amino acid from mussel byssal plaques called 3,4-dihydroxyphenyl--L-alanine, which has been known to generate strong bonding under wet conditions. The photocrosslinking segment consists of an anthracene based monomer used for mechanical fortification of polymer chains. The water soluble segment consists of poly(acrylic acid), which has been known to increase water solubility of polymers and increase adhesion strength of adhesives. The terpolymer was designed to easily applicable using biologically friendly solvents including water and ethanol. Structural design was confirmed by NMR and UV-Vis spectroscopy. Reversible cycloaddition reactions were executed using a handheld UV lamp along with a photoreactor. Molecular weight increases were seen from 4.120 x 104 Da to 7.429 x 104 Da. Lap shear strength testing showed effects of UV exposure through increases in adhesion energy above 450%. Multiple application variables were tested to determine optimal conditions, such as solvent, concentration, and substrate. Currently, optimal conditions show a 1:1 weight ratio of polymer:solvent in water for all surfaces.

  11. Influence of polyvinyl alcohol amount on producing in situ photo-crosslinked thioamide functionalized nanofiber membranes

    Directory of Open Access Journals (Sweden)

    Zeytuncu Bihter


    Full Text Available Poly(vinyl alcohol/maleic anhydride/acryloyl thioamide monomer (PVA/MA/ATM photo-cured nanofiber membranes and pure PVA nanofiber membranes were produced by electrospinning technique. In situ UV radiation was applied during the electrospinning in order to provide polymerization during the jet flight and promote crosslinking of ATM and MA with PVA. The cross-linking was examined by Fourier-transform infrared spectroscopy (FTIR. The morphology and thermal behavior of electrospun nanofiber were characterized by scanning electron microscope (SEM and thermogravimetric analysis (TGA, respectively. The surface area of nanofiber membranes was measured by Brunauer-Emmert-Teller (BET analysis. Furthermore, water durability test was examined. Water durability test demonstrated that in situ photo-cured PVA/MA/ATM nanofiber membrane had the least average mass loss. The surface areas of PVA/MA/ATM nanofiber membranes were 160-280 m2/g. The surface area and diameter of PVA/MA/ATM nanofibers decreased as the PVA content increased. The diameter of nanofibers was obtained less than 100 nm. The results showed that the water-insoluble nanofiber membranes with better chemical and thermal resistance were obtained. These nanofiber membranes may be a promising candidate for the usage of water treatment.

  12. Synthesis and properties of photo-crosslinked mixed-macromer networks

    NARCIS (Netherlands)

    Zant, E.


    Regenerative medicine aims at replacing, engineering or regenerating human cells and tissues. There is a need for novel biomaterials to function as a temporal replacement of damaged tissues or organs. Synthetic biodegradable polymer networks can be such materials, since they offer benefits such as

  13. Fabricating porous, photo-crosslinked poly(trimethylene carbonate) membranes using temperature-induced phase separation

    NARCIS (Netherlands)

    Pasman, Thijs; Grijpma, Dirk W.; Stamatialis, Dimitrios; Poot, Andreas A.


    The recent development of lungs-on-chips is a major advancement in lung disease research. However, the materials used for the membranes in these chips, e.g. poly(dimethyl siloxane) and silicon, are not ideal. This study uses the more biocompatible and mechanically favorable polymer poly(trimethylene

  14. Bioinspired Heparin Nanosponge Prepared by Photo-crosslinking for Controlled Release of Growth Factors

    DEFF Research Database (Denmark)

    Choi, Won Il; Sahu, Abhishek; Vilos, Cristian


    Growth factors have great therapeutic potential for various disease therapy and tissue engineering applications. However, their clinical efficacy is hampered by low bioavailability, rapid degradation in vivo and non-specific biodistribution. Nanoparticle based delivery systems are being evaluated...... factor binding ability. Four different growth factors, bFGF, VEGF, BMP-2, and HGF were successfully encapsulated into Hep-NS. In vitro studies showed sustained release of all the growth factors for almost 60 days and the rate of release was directly dependent on the amount of heparin in Hep......-NS. The released growth factors retained their bioactivity as assessed by a cell proliferation assay. This heparin nanosponge is therefore a promising nanocarrier for the loading and controlled release of growth factors....

  15. Preparation and Mechanical Properties of Photo-Crosslinked Fish Gelatin/Imogolite Nanofiber Composite Hydrogel

    Directory of Open Access Journals (Sweden)

    Mitsuhiro Shibata


    Full Text Available Fish gelatin (FG extracted from sea bream scales was reacted with glycidyl methacrylate (GMA, and the product (FG-GMA was used for photopolymerization using a radical photoinitiator in the presence or absence of imogolite nanofibers in the aqueous solution. The synthesis of FG-GMA was confirmed by 1H NMR spectroscopy, and photopolymerization of FG-GMA was achieved successfully by irradiation with ultraviolet (UV light for 3 min to yield translucent composite hydrogels. The concentration of FG-GMA varied from 10% to 30% without imogolite, and that of imogolite varied from 0% to 2.0%. A microtomed gel sample was observed with a transmission electron microscope (TEM, and imogolite nanofibers were found to be dispersed finely in the gelatin matrix. Scanning electron microscope (SEM observation of the lyophilized gel revealed that it had a porous morphology. Mechanical properties of hydrogels were measured by compression tests using a mechanical tester, and viscoelastic properties were measured using a rheometer. The mechanical strength and storage modulus of the hydrogel increased with an increase of imogolite.

  16. Preparation and Mechanical Properties of Photo-Crosslinked Fish Gelatin/Imogolite Nanofiber Composite Hydrogel (United States)

    Teramoto, Naozumi; Hayashi, Akihiko; Yamanaka, Kaori; Sakiyama, Asako; Nakano, Asuka; Shibata, Mitsuhiro


    Fish gelatin (FG) extracted from sea bream scales was reacted with glycidyl methacrylate (GMA), and the product (FG-GMA) was used for photopolymerization using a radical photoinitiator in the presence or absence of imogolite nanofibers in the aqueous solution. The synthesis of FG-GMA was confirmed by 1H NMR spectroscopy, and photopolymerization of FG-GMA was achieved successfully by irradiation with ultraviolet (UV) light for 3 min to yield translucent composite hydrogels. The concentration of FG-GMA varied from 10% to 30% without imogolite, and that of imogolite varied from 0% to 2.0%. A microtomed gel sample was observed with a transmission electron microscope (TEM), and imogolite nanofibers were found to be dispersed finely in the gelatin matrix. Scanning electron microscope (SEM) observation of the lyophilized gel revealed that it had a porous morphology. Mechanical properties of hydrogels were measured by compression tests using a mechanical tester, and viscoelastic properties were measured using a rheometer. The mechanical strength and storage modulus of the hydrogel increased with an increase of imogolite.

  17. Inactivated human platelet lysate with psoralen: a new perspective for mesenchymal stromal cell production in Good Manufacturing Practice conditions. (United States)

    Castiglia, Sara; Mareschi, Katia; Labanca, Luciana; Lucania, Graziella; Leone, Marco; Sanavio, Fiorella; Castello, Laura; Rustichelli, Deborah; Signorino, Elena; Gunetti, Monica; Bergallo, Massimiliano; Bordiga, Anna Maria; Ferrero, Ivana; Fagioli, Franca


    Mesenchymal stromal cells (MSC) are ideal candidates for regenerative and immunomodulatory therapies. The use of xenogeneic protein-free Good Manufacturing Practice-compliant growth media is a prerequisite for clinical MSC isolation and expansion. Human platelet lysate (HPL) has been efficiently implemented into MSC clinical manufacturing as a substitute for fetal bovine serum (FBS). Because the use of human-derived blood materials alleviates immunologic risks but not the transmission of blood-borne viruses, the aim of our study was to test an even safer alternative than HPL to FBS: HPL subjected to pathogen inactivation by psoralen (iHPL). Bone marrow samples were plated and expanded in α-minimum essential medium with 10% of three culture supplements: HPL, iHPL and FBS, at the same time. MSC morphology, growth and immunophenotype were analyzed at each passage. Karyotype, tumorigenicity and sterility were analyzed at the third passage. Statistical analyses were performed. The MSCs cultivated in the three different culture conditions showed no significant differences in terms of fibroblast colony-forming unit number, immunophenotype or in their multipotent capacity. Conversely, the HPL/iHPL-MSCs were smaller, more numerous, had a higher proliferative potential and showed a higher Oct-3/4 and NANOG protein expression than did FBS-MSCs. Although HPL/iHPL-MSCs exhibit characteristics that may be attributable to a higher primitive stemness than FBS-MSCs, no tumorigenic mutations or karyotype modifications were observed. We demonstrated that iHPL is safer than HPL and represents a good, Good Manufacturing Practice-compliant alternative to FBS for MSC clinical production that is even more advantageous in terms of cellular growth and stemness. Copyright © 2014 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  18. SU-C-204-06: Monte Carlo Dose Calculation for Kilovoltage X-Ray-Psoralen Activated Cancer Therapy (X-PACT): Preliminary Results

    Energy Technology Data Exchange (ETDEWEB)

    Mein, S [Duke University Medical Physics Graduate Program (United States); Gunasingha, R [Department of Radiation Safety, Duke University Medical Center (United States); Nolan, M [Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University (United States); Oldham, M; Adamson, J [Department of Radiation Oncology, Duke University Medical Center (United States)


    promise for accurate quantification of dose for this novel psoralen X-ray therapy. Funding Support, Disclosures, & Conflict of Interest: The Monte Carlo simulation work was not funded; Drs. Adamson & Oldham have received funding from Immunolight LLC for X-PACT research.

  19. Topical or oral administration with an extract of Polypodium leucotomos prevents acute sunburn and psoralen-induced phototoxic reactions as well as depletion of Langerhans cells in human skin

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez, S.; Pathak, M.A.; Fitzpatrick, T.B. [Massachusetts General Hospital, Harvard Medical School, Dept. of Dermatology, Boston, MA (United States); Cuevas, J. [Hospital Universitario de Guadalajara, Dept. of Pathology, Guadalajara (Spain); Villarrubia, V.G. [I.F. Cantabria SA, Medical Dept., Immunology Sect., Madrid (Spain)


    Sunburn, immune suppression, photo-aging, and skin cancers result from uncontrolled overexposure of human skin to solar ultraviolet radiation (UVR). Preventive measures, including photo-protection, are helpful and can be achieved by topical sun-screening agents. Polypodium leucotomos (PL) has been used for the treatment of inflammatory diseases and has shown some in vitro and in vivo immunomodulating properties. Its beneficial photo-protective effects in the treatment of vitiligo and its antioxidant properties encouraged us to evaluate in vivo the potentially useful photo-protective property of natural extract of PL after topical application or oral ingestion. Twenty-one healthy volunteers [either untreated or treated with oral psoralens (8-MOP or 5-MOP)] were enrolled in this study and exposed to solar radiation for evaluation of the following clinical parameters: immediate pigment darkening (IPD), minimal erythema dose (MED), minimal melanogenic dose (MMD), and minimal phototoxic dose (MPD) before and after topical or oral administration of PL. Immunohistochemical assessment of CD1a-expressing epidermal cells were also performed. PL was found to be photo-protective after topical application as well as oral administration. PL increased UV dose required for IPD (P<0.01), MED (P<0.001) and MPD (P<0.001). After oral administration of PL, MED increased 2.,8{+-}0.59 times and MPD increased 2.75{+-}0.5 and 6.8{+-}1.3 times depending upon the type of psoralen used. Immunohistochemical study revealed photo-protection of Langherhans cells by oral as well as topical PL. The observed photo-protective activities of oral or topical PL reveal a new avenue in examining the potentially useful field of systemic photo-protection and suggests that PL can be used as adjunct treatment and can make photochemotherapy and phototherapy possibly safe and effective when the control of cutaneous phototoxicity to PUVA or UVB is a limiting factor in such photo-therapies. (au). 50 refs.

  20. Fluorescence spectroscopy for non-invasive measurement of mechanical stiffness after photo-crosslinking of rabbit cornea (United States)

    Williams, Maura; Lewis, William; Ortega-Martinez, Antonio; Franco, Walfre


    Background and Objectives: Keratoconus is a disease characterized by progressive steepening and thinning of the cornea, altering visual acuity and sometimes potentiating the need for corneal transplant if the disease progresses.1-3 Corneal crosslinking, a procedure that uses topical riboflavin and UV light to increase the stiffness of the cornea through the creation of collagen crosslinks was recently approved by the FDA for use in the U.S. The objective of the present study was to investigate whether endogenous collagen fluorescence changes following treatment can be correlated to alterations in the stiffness of the cornea, thereby guiding treatment parameters. Study Design and Results: 78 ex-vivo rabbit eyes divided into three groups: riboflavin solution plus UV irradiation, dextran solution plus UV irradiation, and riboflavin solution only. An additional group of eyes received no treatment. The epithelium was removed from each sample and topical riboflavin was applied. Eyes were irradiated with a 365 nm black ray UV lamp for various treatment times, ranging from half the clinical treatment time to three times the length. Mechanical testing was performed to determine the force/displacement relationship for the various treatment times. Fluorescence spectral changes following treatment corresponded with changes in stiffness. In particular, a decrease in the value of fluorescence intensity at 290/340 nm excitation/emission wavelengths corresponded to an increase in corneal stiffness following treatment. It may be possible to use fluorescence spectral changes of endogenous corneal crosslinks to evaluate mechanical stiffness changes non-invasively.

  1. Photo-Crosslinking of Pendent Uracil Units Provides Supramolecular Hole Injection/Transport Conducting Polymers for Highly Efficient Light-Emitting Diodes

    Directory of Open Access Journals (Sweden)

    Hsi-Kang Shih


    Full Text Available A new process for modifying a polymeric material for use as a hole injection transport layer in organic light-emitting diodes has been studied, which is through 2π + 2π photodimerization of a DNA-mimetic π-conjugated poly(triphenylamine-carbazole presenting pendent uracil groups (PTC-U under 1 h of UV irradiation. Multilayer florescence OLED (Organic light-emitting diodes device with the PTC-U-1hr as a hole injection/transport layer (ITO (Indium tin oxide/HITL (hole-injection/transport layer (15 nm/N,N'-di(1-naphthyl- N,N'-diphenyl-(1,1'-biphenyl-4,4'-diamine (NPB (15 nm/Tris-(8-hydroxyquinoline aluminum (Alq3 (60 nm/LiF (1 nm/Al (100 nm is fabricated, a remarkable improvement in performance (Qmax (external quantum efficiency = 2.65%, Bmax (maximum brightness = 56,704 cd/m2, and LE (luminance efficiencymax = 8.9 cd/A relative to the control PTC-U (Qmax = 2.40%, Bmax = 40,490 cd/m2, and LEmax = 8.0 cd/A. Multilayer phosphorescence OLED device with the PTC-U-1hr as a hole injection/transport layer (ITO/HITL (15 nm/Ir(ppy3:PVK (40 nm/BCP (10nm/Alq3 (40 nm/LiF (1 nm/Al (100 nm is fabricated by successive spin-coating processes, a remarkable improvement in performance (Qmax = 9.68%, Bmax = 41,466 cd/m2, and LEmax = 36.6 cd/A relative to the control PTC-U (Qmax = 8.35%, Bmax = 34,978 cd/m2, and LEmax = 30.8 cd/A and the commercial product (poly(3,4-ethylenedioxythiophene:polystyrenesulfonate PEDOT:PSS (Qmax = 4.29%, Bmax = 15,678 cd/m2, and LEmax = 16.2 cd/A has been achieved.

  2. Micro-porous composite scaffolds of photo-crosslinked poly(trimethylene carbonate) and nano-hydroxyapatite prepared by low-temperature extrusion-based additive manufacturing

    NARCIS (Netherlands)

    Geven, Mike A.; Sprecher, Christoph; Guillaume, Olivier; Eglin, David; Grijpma, Dirk W.


    Complex bony defects such as those of the orbital floor are challenging to repair. Additive manufacturing techniques open up possibilities for the fabrication of implants with a designed macro-porosity for the reconstruction of such defects. Apart from a designed macro-porosity for tissue ingrowth,

  3. Unnatural amino acid photo-crosslinking of the IKs channel complex demonstrates a KCNE1:KCNQ1 stoichiometry of up to 4:4. (United States)

    Murray, Christopher I; Westhoff, Maartje; Eldstrom, Jodene; Thompson, Emely; Emes, Robert; Fedida, David


    Cardiac repolarization is determined in part by the slow delayed rectifier current (IKs), through the tetrameric voltage-gated ion channel, KCNQ1, and its β-subunit, KCNE1. The stoichiometry between α and β-subunits has been controversial with studies reporting either a strict 2 KCNE1:4 KCNQ1 or a variable ratio up to 4:4. We used IKs fusion proteins linking KCNE1 to one (EQ), two (EQQ) or four (EQQQQ) KCNQ1 subunits, to reproduce compulsory 4:4, 2:4 or 1:4 stoichiometries. Whole cell and single-channel recordings showed EQQ and EQQQQ to have increasingly hyperpolarized activation, reduced conductance, and shorter first latency of opening compared to EQ - all abolished by the addition of KCNE1. As well, using a UV-crosslinking unnatural amino acid in KCNE1, we found EQQQQ and EQQ crosslinking rates to be progressively slowed compared to KCNQ1, which demonstrates that no intrinsic mechanism limits the association of up to four β-subunits within the IKs complex.

  4. Comparative profiling of biomarker psoralen in antioxidant active ...

    African Journals Online (AJOL)

    Materials and Methods: HPTLC studies were carried out using CAMAG HPTLC system on Glass-backed silica gel 60F254 HPTLC pre-coated plates using selected mobile phase toluene: methanol (9:1). The antioxidant activity was carried out, using DPPH free radical method. Results: Among all the five species of genus ...

  5. Treatment of necrobiosis lipoidica with topical psoralen plus ultraviolet A

    NARCIS (Netherlands)

    de Rie, M. A.; Sommer, A.; Hoekzema, R.; Neumann, H. A. M.


    Background Necrobiosis lipoidica (NL) is a rare skin disease, mostly seen on the legs and often occurring in patients with diabetes mellitus. The disease belongs to the idiopathic cutaneous palisading granulomatous dermatitides associated with a degeneration of collagen, thus leading to skin

  6. Spectral effects in activation of the human immunodeficiency virus promoter by psoralens plus ultraviolet A treatment. (United States)

    Miller, S A; Beer, J Z; Strickland, A G; Zmudzka, B Z

    The effects of PUVA treatment on HIV promoter activation and cell killing in HIV cat/HeLa cells were studied using two UV sources, a UVASUN sunlamp and a UVAR Photoactivation Chamber. A 4 to 5 times higher dose of ultraviolet radiation was required from the UVASUN lamp than from the UVAR lamps: 1) to activate the HIV promoter in the presence of 0.1 or 1.0 microgram/ml 8-MOP and 2) to reduce cell survival to a level of 10%, in the presence of 0.1 or 1.0 microgram/ml 8-MOP. In addition, exposures performed with a fixed dose of 20 kJ/m2 at varying concentrations of 8-MOP, required a 4.7 times higher combined PUVA dose from the UVASUN lamp than from the UVAR lamps. Two possible sources of these differences were analyzed: (1) the presence of UVB + UVA2 (280-340 nm) in the radiation emitted by the UVAR, but not the UVASUN lamp, and its potential biological activity independent of 8-MOP, and (2) the difference in the overlap of the emission spectra of the two lamps with the absorption spectrum of 8-MOP. The area of overlap was higher for the UVAR lamp than for the UVASUN lamp by a factor of 4.6, which is close to the difference between these two lamps in induction of the HIV promoter and killing HeLa cells. This indicates that the effectiveness of a particular UVA source used in combination with 8-MOP can be predicted by its congruence to the absorption spectrum of the photosensitizing drug.

  7. UVB-activated psoralen reduces luminal narrowing after balloon dilation because of inhibition of constrictive remodeling

    NARCIS (Netherlands)

    Perrée, Jop; van Leeuwen, Ton G.; Velema, Evelyn; Smeets, Mirjam; de Kleijn, Dominique; Borst, Cornelius


    In this study we have explored the potential of PUVB (8-MOP + UVB) therapy for the reduction of luminal narrowing after arterial injury. In 15 rabbits, balloon dilation of iliac arteries was performed. In 20 arteries, dilation was combined with the delivery of pulsed ultraviolet light B (UVB)

  8. Determination of the minimal phototoxic dose and colorimetry in psoralen plus ultraviolet A radiation therapy. (United States)

    Kraemer, Cristine Kloeckner; Menegon, Dóris Baratz; Cestari, Tania Ferreira


    The use of an adequate initial dose of ultraviolet A (UVA) radiation for photochemotherapy is important to prevent burns secondary to overdosage, meanwhile avoiding a reduced clinical improvement and long-term risks secondary to underdosage. The ideal initial dose of UVA can be achieved based on the phototype and the minimal phototoxic dose (MPD). The objective measurement of constitutive skin color (colorimetry) is another method commonly used to quantify the erythematous skin reaction to ultraviolet radiation exposure. The aim of this study was to determine variations in MPD and constitutional skin color (coordinate L(*)) within different phototypes in order to establish the best initial dose of UVA radiation for photochemotherapy patients. Thirty-six patients with dermatological conditions and 13 healthy volunteers were divided into five groups according to phototype. Constitutional skin color of the infra-axillary area was assessed by colorimetry. The infra-axillary area was subsequently divided into twelve 1.5 cm(2) regions to determine the MPD; readings were performed 72 h after oral administration of 8-methoxypsoralen (MOP) followed by exposure of the demarcated regions to increasing doses of UVA. The majority of the participants were women (73.5%) and their mean age was 38.8 years. The MPD ranged from 4 to 12 J/cm(2) in phototypes II and III; from 10 to 18 J/cm(2) in type IV; from 12 to 24 J/cm(2) in type V and from 18 to 32 J/cm(2) in type VI. The analysis of colorimetric values (L(*) coordinate) and MPD values allowed the definition of three distinctive groups of individuals composed by phototypes II and III (group 1), types IV and V (group 2), and phototype VI (group 3). MPD and L(*) coordinate showed variation within the same phototype and superposition between adjacent phototypes. The values of the L(*) coordinate and the MPD lead to the definition of three distinct sensitivity groups: phototypes II and III, IV and V and type VI. Also, the MPD values bear a strong correlation to coordinate L(*) values. Mean MPD values described for each of the three major sensitivity groups were higher than the values commonly used in clinical settings for the different phototypes. Therefore, phototype alone is not a good parameter to define the initial UVA dose. MPD and colorimetry could be used in pre-treatment evaluation of patients who are to be submitted to photochemotherapy, in a non-invasive and more accurate way when compared with the classical phenotype clinical criteria.

  9. Interplay between human high mobility group protein 1 and replication protein A on psoralen-cross-linked DNA

    DEFF Research Database (Denmark)

    Reddy, Madhava C; Christensen, Jesper; Vasquez, Karen M


    Human high mobility group box (HMGB) 1 and -2 proteins are highly conserved and abundant chromosomal proteins that regulate chromatin structure and DNA metabolism. HMGB proteins bind preferentially to DNA that is bent or underwound and to DNA damaged by agents such as cisplatin, UVC radiation...

  10. Human XPC-hHR23B interacts with XPA-RPA in the recognition of triplex-directed psoralen DNA interstrand crosslinks

    DEFF Research Database (Denmark)

    Thoma, Brian S; Wakasugi, Mitsuo; Christensen, Jesper


    DNA interstrand crosslinks (ICLs) represent a severe form of damage that blocks DNA metabolic processes and can lead to cell death or carcinogenesis. The repair of DNA ICLs in mammals is not well characterized. We have reported previously that a key protein complex of nucleotide excision repair (...

  11. Photo-inducible crosslinked nanoassemblies for pH-controlled drug release. (United States)

    Dickerson, Matthew; Winquist, Nickolas; Bae, Younsoo


    To control drug release from block copolymer nanoassemblies by variation in the degree of photo-crosslinking and inclusion of acid sensitive linkers. Poly(ethylene glycol)-poly(aspartate-hydrazide-cinnamate) (PEG-CNM) block copolymers were prepared and conjugated with a model drug, doxorubicin (DOX), through acid sensitive hydrazone linkers. The block copolymers formed photo-inducible, self-assembled nanoassemblies (piSNAs), which were used to produce photo-inducible crosslinked nanoassemblies (piCNAs) through UV crosslinking. The nanoassemblies were characterized to determine particle size, surface charge, pH- and crosslinking-dependent DOX release, in vitro cytotoxicity, and intracellular uptake as a function of photo-crosslinking degree. Nanoassemblies with varying photo-crosslinking degrees were successfully prepared while retaining particle size and surface charge. Photo-crosslinking caused no noticeable change in DOX release from the nanoassemblies at pH 7.4, but the DOX-loaded nanoassemblies modulated drug release as a function of crosslinking at pH 6.0. The nanoassemblies showed similar cytotoxicity regardless of crosslinking degrees, presumably due to the low cellular uptake and cell nucleus drug accumulation. Photo-crosslinking is useful to control drug release from pH-sensitive block copolymer nanoassemblies as a function of crosslinking without altering the particle properties, and thus providing unique tools to investigate the pharmaceutical effects of drug release on cellular response.

  12. Simultaneous determination of scopoletin, psoralen, bergapten, xanthotoxin, columbianetin acetate, imperatorin, osthole and isoimperatorin in rat plasma by LC-MS/MS for pharmacokinetic studies following oral administration of Radix Angelicae Pubescentis extract. (United States)

    Chang, Yan-xu; Zhang, Qiu-Hong; Li, Jin; Zhang, Ling; Guo, Xin-rong; He, Jun; Zhang, Peng; Ma, Lin; Deng, Yan-ru; Zhang, Bo-li; Gao, Xiu-mei


    A rapid and sensitive bioassay based on liquid chromatography tandem mass spectrometry (LC-MS/MS) has been developed and validated for the simultaneous determination of eight coumarins in rat plasma. The liquid-liquid extraction method with ethyl acetate was used to prepare the plasma samples after addition of warfarin as an internal standard (IS). Chromatographic separation was performed on an Eclipse plus C18 column (100mm×4.6mm, 1.8μm) using gradient elution when 1mM ammonium acetate aqueous solution - acetonitrile was used as the mobile phase. The lower limit of quantitation (LLOQ) of each coumarin was lower than 2.16ngmL(-1). Intra-day and inter-day precisions were less than 15%. The accuracies were in the range of 88.9-117%. The mean recoveries of coumarins and IS were higher than 84%. The method was successfully applied to a pharmacokinetic study of eight coumarins in rats after oral administration of radix angelicae pubescentis. Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.

  13. A poly(D,L-lactide) resin for the preparation of tissue engineering scaffolds by stereolithography

    NARCIS (Netherlands)

    Melchels, Ferry P. W.; Feijen, Jan; Grijpma, Dirk W.

    Porous polylactide constructs were prepared by stereolithography, for the first time without the use of reactive diluents. Star-shaped poly(D,L-lactide) oligomers with 2, 3 and 6 arms were synthesised, end-functionalised with methacryloyl chloride and photo-crosslinked in the presence of ethyl

  14. Biocomposites of cellulose reinforced starch: improvement of properties by photo-induced crosslinking. (United States)

    Kumar, Annamalai Pratheep; Singh, Raj Pal


    In the present study, the composite films have been prepared from the aqueous dispersions of starch with microcrystalline cellulose using glycerol as plasticizer and irradiated under ultraviolet (UV) light using sodium benzoate as photo-sensitizer. Photo-crosslinking was characterized by measuring the water absorption under 100% relative humidity, swelling degree and gel fraction in dimethylsulphoxide (DMSO), upon irradiation time. Both, the incorporation of cellulose and photo-irradiation were found to decrease the water absorption, swelling in DMSO and increase the gel fraction. Thermal transitions indicated the anti-plasticization of amylopectin chains at the fiber/matrix interface. With increasing content of cellulose and photo-irradiation time, the tensile modulus and strength were found to improve. It is summarized that the combination of cellulose reinforcement and photo-crosslinking of matrix has improved the physical and mechanical properties.

  15. Phototoxicity activity of Psoralea drupacea L. using Atremia salina bioassay system

    Directory of Open Access Journals (Sweden)

    Mohammad Ramezani


    Conclusion: The result showed that P. drupacea methanolic extract and chloroform fraction have phototoxicity in A. salina bioassay system and their toxic effect is related to phototoxic constituents such as psoralen.

  16. Disease: H01663 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available o be first-line therapies for those with GPP. Adalimumab, etanercept, and psoralen plus ultraviolet A are second-line moda...rms alter which agents are first-line modalities as concerns such as teratogenici

  17. Concurrent Stabilization and Imaging of a Novel Polymer for Second Harmonic Generation via In Situ Photopolymerization (United States)


    neither photochemistry is compatible with the desirable chromophores that are based on stilbene or azobenzene moieties. Our goal is to such as the curing of diepoxy- diamine systems,5 -9 ) diisocyanate-diol (or triol), 10-13) or diisocyanate-diaminel 4 ) systems, or of...the photo- crosslinking step often causes a direct or sensitized trans to cis isomerization of stilbene or azobenzene based NLO chromophores

  18. Human MLH1 protein participates in genomic damage checkpoint signaling in response to DNA interstrand crosslinks, while MSH2 functions in DNA repair.

    Directory of Open Access Journals (Sweden)

    Qi Wu


    Full Text Available DNA interstrand crosslinks (ICLs are among the most toxic types of damage to a cell. For this reason, many ICL-inducing agents are effective therapeutic agents. For example, cisplatin and nitrogen mustards are used for treating cancer and psoralen plus UVA (PUVA is useful for treating psoriasis. However, repair mechanisms for ICLs in the human genome are not clearly defined. Previously, we have shown that MSH2, the common subunit of the human MutSalpha and MutSbeta mismatch recognition complexes, plays a role in the error-free repair of psoralen ICLs. We hypothesized that MLH1, the common subunit of human MutL complexes, is also involved in the cellular response to psoralen ICLs. Surprisingly, we instead found that MLH1-deficient human cells are more resistant to psoralen ICLs, in contrast to the sensitivity to these lesions displayed by MSH2-deficient cells. Apoptosis was not as efficiently induced by psoralen ICLs in MLH1-deficient cells as in MLH1-proficient cells as determined by caspase-3/7 activity and binding of annexin V. Strikingly, CHK2 phosphorylation was undetectable in MLH1-deficient cells, and phosphorylation of CHK1 was reduced after PUVA treatment, indicating that MLH1 is involved in signaling psoralen ICL-induced checkpoint activation. Psoralen ICLs can result in mutations near the crosslinked sites; however, MLH1 function was not required for the mutagenic repair of these lesions, and so its signaling function appears to have a role in maintaining genomic stability following exposure to ICL-induced DNA damage. Distinguishing the genetic status of MMR-deficient tumors as MSH2-deficient or MLH1-deficient is thus potentially important in predicting the efficacy of treatment with psoralen and perhaps with other ICL-inducing agents.

  19. Biotransformation of furanocoumarins by Cunninghamella elegans


    Attia, Ghada Ismail El-shahat Ali; Kamillia A. Abou-El-seoud; Ibrahim, Abdel-Rahim Sayed


    Biotransformation of Furanocoumarins; psoralen (1), bergapten (2), xanthotoxin (3) and imperatorin (4) was explored by Cunninghamella elegans NRRL 1392, revealing the metabolism of psoralen (1) and bergapten (2) into bergaptol (5), while xanthotoxin (3) and imperatorin (4) were converted into xanthotoxol (6). On the other hand unexpected conversion of xanthotoxin (3) into 3,4 dihydroxanthotoxin (7) occurred. The structure of the isolated pure metabolites was established using physical and spe...

  20. Alginate- and gelatin-based bioactive photocross-linkable hybrid materials for bone tissue engineering. (United States)

    Lewandowska-Łańcucka, Joanna; Mystek, Katarzyna; Mignon, Arn; Van Vlierberghe, Sandra; Łatkiewicz, Anna; Nowakowska, Maria


    The paper presents the synthesis, the physico-chemical and the biological properties of novel hybrid materials prepared from photo-crosslinked gelatin/alginate-based hydrogels and silica particles exhibiting potential for the regeneration of bone tissue. Both alginate and gelatin were functionalized with methacrylate and methacrylamide moieties, respectively to render them photo-crosslinkable. Submicron silica particles of two sizes were dispersed within three types of polymeric sols including alginate, gelatin, and gelatin/alginate blends, which were subsequently photo-crosslinked. The swelling ratio, the gel fraction and the mechanical properties of the hybrid materials developed were examined and compared to these determined for reference hydrogel matrices. The in vitro cell culture studies have shown that the prepared materials exhibited biocompatibility as they supported both MEFs and MG-63 mitochondrial activity. Finally, the in vitro experiments performed under simulated body fluid conditions have revealed that due to inclusion of silica particles into the biopolymeric hydrogel matrices the mineralization was successfully induced. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Phototherapy and photopharmacology

    Energy Technology Data Exchange (ETDEWEB)

    Gasparro, F.P.; Chan, G.; Edelson, R.L.


    The activation of 8-methoxypsoralen (8-MOP) by long-wavelength ultraviolet A light (UVA, 320-400 nm) induces the formation of interstrand cross-links in DNA. Psoralen plus UVA (PUVA) is widely used in the treatment of psoriasis, a hyperproliferative disease of the skin. A new psoralen plus UVA therapy has been developed in which the 8-MOP-containing blood of cutaneous T cell lymphoma (CTCL) patients is irradiated with UVA light extracorporeally (i.e., extracorporeal photopheresis). The first group of patients had the leukemic variant of CTCL. A regimen of two treatments on successive days at monthly intervals produced a clinical response in eight of 11 patients. In this review the properties of several psoralens (both naturally occurring and synthetic derivatives) are compared, using several assays (DNA cross-linking, inhibition of lymphocyte response to mitogen stimulation, and cell viability). The development of a panel of monoclonal antibodies that recognize 8-MOP-modified DNA is also described. These antibodies have been used to quantitate 8-MOP photoadduct levels in human DNA samples. In addition to the psoralens, the light activation of two other compounds, gilvocarcin and an insulin-psoralen conjugate, is described.

  2. Extensive burns caused by the abusive use of photosensitizing agents. (United States)

    Braye, F; Latarjet, J; Foyatier, J L; Comparin, J P; Tranchand, P; Boucaud, C


    Psoralens are photosensitizing agents used in dermatology as reinforcements in psoralen ultraviolet A-range therapy. We report observations of 14 young women hospitalized for severe burns caused by abusive use of psoralens. The burns were of superficial and deep second-degree depth and covered more than 76% of the body surface on average. All patients needed fluid resuscitation. Hospital stay was 11 days on average. Healing was obtained without skin grafting in all cases. Among the six patients who responded to the mailed questionnaire, negative effects are now present in all patients as inflammatory peaks. Two patients have esthetic sequelae such as dyschromia and scars. The misuse of photosensitizing agents poses many problems. These accidents are very expensive. The largeness of the burned surface can involve a fatal prognosis. And finally, one can suspect that a much larger portion of the population regularly uses these products without any serious accident. In this case carcinogenesis can be expected.

  3. Silicon-Containing Polymers and Composites (United States)


    coated PEGDA + POSS films Pure PEGDA Can hydrogen bond with water Photo-crosslinkable Fluorodecyl POSS molecules Si Si Si O O O Si OO ORf R Rf Rf...and intermolecular hydrogen bonding between silanols. • Mr=,monoclinic, space group P2(1)/c , a=11.84(10) Å, b=57.11(6) Å, c=19.06(2) Å...release; distribution unlimited. Rf Rf R = CH2CH2(CF2)7CF3 -17.9 ppm-17.1 ppm-45.5 ppm Initial Copolymerizations MMA (MW = 100 g/mol) MMA -F-POSS (MW

  4. Cell-induced flow-focusing instability in gelatin methacrylate microdroplet generation


    Jung, Jinmu; Oh, Jonghyun


    Photo-crosslinkable gelatin methacrylate (GelMa) microspheres are applicable to deliver cells or drugs in biological or biomedical applications. To fabricate GelMa microdroplets, a flow focusing technique with advantages of size control and rapid production was used in a T-junction microfluidic device. Instability played an important role in promoting microdroplet uniformity. 5 wt. % GelMa prepolymer solution mixed with cells affected cell-induced instability. At low flow rate ratio of GelMa ...

  5. Electron microscopy methods for studying in vivo DNA replication intermediates. (United States)

    Lopes, Massimo


    The detailed understanding of the DNA replication process requires structural insight. The combination of psoralen crosslinking and electron microscopy has been extensively exploited to reveal the fine architecture of in vivo DNA replication intermediates. This approach proved instrumental to uncover the basic mechanisms of DNA duplication, as well as the perturbation of this process by genotoxic treatments. The replication structures need to the stabilized in vivo (by psoralen crosslinking) prior to extraction and enrichment procedures, finally leading to the visualization at the transmission electron microscope. This chapter outlines the procedures required to visualize in vivo replication intermediates of genomic DNA, extracted from budding yeast or cultured mammalian cells.

  6. Biotransformation of furanocoumarins by Cunninghamella elegans

    Directory of Open Access Journals (Sweden)

    Ghada Ismail El-shahat Ali Attia


    Full Text Available Biotransformation of Furanocoumarins; psoralen (1, bergapten (2, xanthotoxin (3 and imperatorin (4 was explored by Cunninghamella elegans NRRL 1392, revealing the metabolism of psoralen (1 and bergapten (2 into bergaptol (5, while xanthotoxin (3 and imperatorin (4 were converted into xanthotoxol (6. On the other hand unexpected conversion of xanthotoxin (3 into 3,4 dihydroxanthotoxin (7 occurred. The structure of the isolated pure metabolites was established using physical and spectroscopic techniques including, melting points, IR, 1H NMR, 13C NMR and mass spectroscopy.

  7. Phototherapeutic, photobiologic, and photosensitizing properties of khellin

    Energy Technology Data Exchange (ETDEWEB)

    Morliere, P.; Hoenigsmann, H.A.; Averbeck, D.; Dardalhon, M.; Hueppe, G.O.; Ortel, B.; Santus, R.; Dubertret, L.


    Khellin, whose chemical structure closely resembles that of psoralen, is reported to be an efficient drug for treating vitiligo when combined with ultraviolet A irradiation. Photobiological activity on yeast is found to be much lower than that of bifunctional psoralens such as 5-methoxypsoralen. In vitro experiments reveal that khellin is a poor photosensitizer. It behaves as a monofunctional agent with respect to DNA photoaddition. It does not photoinduce cross-links in DNA in vitro or in Chinese hamster cells in vivo. This behavior may explain the low photogenotoxicity in yeast and the lack of phototoxic erythemal response when treating vitiligo with khellin.

  8. Photo(chemotherapy for Vitiligo

    Directory of Open Access Journals (Sweden)

    Güzin Özarmağan


    Full Text Available Vitiligo is a skin disease having an impact on the quality of life. Phototherapy or photochemotherapy are the first-line treatment choices in generalized vitiligo and are among the most effective treatment modalities either alone or in combination with other therapies. Treatment methods with ultraviolet include oral / topical psoralen plus UVA (PUVA, psoralen plus sunlight (PUVAsol, UVA, broadband UVB, narrowband UVB, excimer laser, monochromatic excimer light and sunlight. This review focuses on the modalities, principals and studies of UV treatment in vitiligo.

  9. Design of triple-shape memory polyurethane with photo-cross-linking of cinnamon groups. (United States)

    Wang, Lin; Yang, Xifeng; Chen, Hongmei; Gong, Tao; Li, Wenbing; Yang, Guang; Zhou, Shaobing


    A triple-shape memory polyurethane (TSMPU) with poly(ε-caprolactone) -diols (PCL-diols) as the soft segments and diphenyl methane diisocyanate (MDI), N,N-bis (2-hydroxyethyl) cinnamamide (BHECA) as the hard segments was synthesized via simple photo-crosslinking of cinnamon groups irradiated under λ > 280 nm ultraviolet (UV) light. Fourier transform infrared spectroscopy (FT-IR), proton nuclear magnetic resonance ((1)H-NMR) and ultraviolet-visible absorption spectrum (UV-vis) confirmed the chemical structure of the material. Differential scanning calorimetry (DSC) and dynamic mechanical analysis (DMA) results demonstrated that the photo-crosslinked polymer possessed two transition temperatures, one is due to the melting point of the soft segment PCL-diols, and the other is due to the glass transition temperature. All these contributed to the cross-linked structure of the hard segments and resulted in an excellent triple-shape memory effect. Alamar blue assay showed that the material has good non-cytotoxicity and can be potentially used in biomaterial devices.

  10. NJP VOLUME 41 NO 4

    African Journals Online (AJOL)



    Jul 24, 2014 ... autism, urticaria, mood disorder, β thalassemia and poly- dactyly4,6,11 but none of these associations have been consistent. There was no similar or novel association in our patient. Treatment of DSH has included the use of topical steroids, pimerolimus, calcipotriol and Psoralen plus UVA (PUVA) all of ...

  11. Ultraviolet A1 phototherapy for mycosis fungoides

    DEFF Research Database (Denmark)

    Olek-Hrab, K; Silny, W; Dańczak-Pazdrowska, A


    Background.  Mycosis fungoides (MF) is the most common form of primary cutaneous lymphoma, and is characterized by a malignant proliferation of CD4+ cells. Psoralen ultraviolet A (PUVA) irradiation is the most common treatment for cutaneous lesions. However, PUVA carries the risk of adverse...

  12. Xi et al., Afr J Tradit Complement Altern Med. (2014) 11(4):94-100 ...

    African Journals Online (AJOL)


    CHBGF on improving life quality in terms of survival time, pathology of tumor tissue and ameliorating peripheral blood cells in mouse ... pathology; peripheral blood cell; H22 hepatoma carcinoma cell. Introduction ..... An experiment study on antitumor activity of Psoralen on mammary cancer cell line EMT6 in vitro and in vivo.

  13. Mismatch repair and nucleotide excision repair proteins cooperate in the recognition of DNA interstrand crosslinks (United States)

    Zhao, Junhua; Jain, Aklank; Iyer, Ravi R.; Modrich, Paul L.; Vasquez, Karen M.


    DNA interstrand crosslinks (ICLs) are among the most cytotoxic types of DNA damage, thus ICL-inducing agents such as psoralen, are clinically useful chemotherapeutics. Psoralen-modified triplex-forming oligonucleotides (TFOs) have been used to target ICLs to specific genomic sites to increase the selectivity of these agents. However, how TFO-directed psoralen ICLs (Tdp-ICLs) are recognized and processed in human cells is unclear. Previously, we reported that two essential nucleotide excision repair (NER) protein complexes, XPA–RPA and XPC–RAD23B, recognized ICLs in vitro, and that cells deficient in the DNA mismatch repair (MMR) complex MutSβ were sensitive to psoralen ICLs. To further investigate the role of MutSβ in ICL repair and the potential interaction between proteins from the MMR and NER pathways on these lesions, we performed electrophoretic mobility-shift assays and chromatin immunoprecipitation analysis of MutSβ and NER proteins with Tdp-ICLs. We found that MutSβ bound to Tdp-ICLs with high affinity and specificity in vitro and in vivo, and that MutSβ interacted with XPA–RPA or XPC–RAD23B in recognizing Tdp-ICLs. These data suggest that proteins from the MMR and NER pathways interact in the recognition of ICLs, and provide a mechanistic link by which proteins from multiple repair pathways contribute to ICL repair. PMID:19468048

  14. Evaluation of Sclerotinia sclerotiorum as a potential mycotoxin producer on soybeans. (United States)

    Ciegler, A; Burbridge, K A; Ciegler, J; Hesseltine, C W


    Solvent extracts of Sclerotinia sclerotiorum sclerotia were nontoxic to mice and chicken embryos; psoralens were not detected. Solvent extracts of soybeans inoculated with 10 strains of S. sclerotiorum were toxic on injection but nontoxic on per os administration to mice. The presence of chlorinated hydrocarbons in the soybeans may partially help explain toxicity by intraperitineal injection. PMID:569464

  15. Liquid crystalline networks for electroluminescent displays

    CERN Document Server

    Contoret, A E A


    This work presents the first low molar mass organic electroluminescent (EL) material to form a nematic glass and then emit plane-polarised light from the vitrified state on application of an electric field. Photocrosslinkable molecules are also discussed which form insoluble films on illumination with ultra-violet light. This approach combines the ease of deposition of small molecules with the robustness and stability of polymers, allowing simple fabrication of multi-layer EL devices and photo-patterning. A range of conjugated low molar-mass molecules are considered, containing the anthracene, perylene and fluorene cores, with the aims of producing a general recipe for efficient EL, based on ordered, stable nematics at room temperature. Many physical properties are compared and molecular mechanics modeling is used to represent molecular geometries. An acrylate and several diene photo-polymerisable derivatives of the fluorenes undergo photo-crosslinking. Infrared and photoluminescence spectroscopy is used to e...

  16. Engineering of filamentous bacteriophage for protein sensing (United States)

    Brasino, Michael

    Methods of high throughput, sensitive and cost effective quantification of proteins enables personalized medicine by allowing healthcare professionals to better monitor patient condition and response to treatment. My doctoral research has attempted to advance these methods through the use of filamentous bacteriophage (phage). These bacterial viruses are particularly amenable to both genetic and chemical engineering and can be produced efficiently in large amounts. Here, I discuss several strategies for modifying phage for use in protein sensing assays. These include the expression of bio-orthogonal conjugation handles on the phage coat, the incorporation of specific recognition sequences within the phage genome, and the creation of antibody-phage conjugates via a photo-crosslinking non-canonical amino acid. The physical and chemical characterization of these engineered phage and the results of their use in modified protein sensing assays will be presented.

  17. Crosslinking poly(acrylic glycidyl ether) honeycomb film by cationic photopolymerization and its converting to inorganic SiO2 film (United States)

    Zheng, Kaijing; Hu, Dongdong; Deng, Yufei; Maitloa, Inamullah; Nie, Jun; Zhu, Xiaoqun


    Highly ordered hexagonal patterns of homopolymer poly(glycidyl methacrylate) (PGMA) thin film was successfully fabricated via a one-step static breath figure process. The regularity and pore size of the films can be modulated by changing solvent composition or polymer concentration. The stable PGMA honeycomb film were formed via cationic photopolymerization due to the epoxide group on the side chain of PGMA. After photo-crosslinked, not only the patterned structures were well preserved, but also the solvent resistance as well as thermal stability of the film was significantly improved. The obtained film could be used as template to assemble SiO2 particles via chemical vapor deposition at mild condition. Under high temperature, the honeycomb film was burned up and the silica particles were sintered into SiO2 microbowl arrays, which could be tunable by varying the structure of the obtained template.

  18. Visualization and interpretation of eukaryotic DNA replication intermediates in vivo by electron microscopy. (United States)

    Neelsen, Kai J; Chaudhuri, Arnab Ray; Follonier, Cindy; Herrador, Raquel; Lopes, Massimo


    The detailed understanding of the DNA replication process requires structural insight. The combination of psoralen cross-linking and electron microscopy has been extensively exploited to reveal the fine architecture of in vivo DNA replication intermediates. This approach proved instrumental to uncover the basic mechanisms of DNA duplication, as well as the perturbation of this process by various forms of replication stress. The replication structures are stabilized in vivo (by psoralen cross-linking) prior to extraction and enrichment procedures, allowing their visualization at the transmission electron microscope. This chapter outlines the procedures required to visualize and interpret in vivo replication intermediates of genomic DNA, extracted from budding yeast, Xenopus egg extracts, or cultured mammalian cells.

  19. STAT3/5-dependent IL9 overexpression contributes to neoplastic cell survival in mycosis fungoides

    DEFF Research Database (Denmark)

    Vieyra-Garcia, Pablo A.; Wei, Tianling; Naym, David Gram


    preparations. To explore the mechanism of IL9 secretion, we knocked down STAT3/5 and IRF4 by siRNA transfection in CTCL cell lines receiving psoralen+UVA (PUVA) ± anti-IL9 antibody. To further examine the role of IL9 in tumor development, the EL-4 T-cell lymphoma model was used in C57BL/6 mice.  Results...

  20. [Effective Components of three kinds of shen-supplementing Chinese medicine on self-renewal and neuron-like differentiation of NSCs in AD mouse embryos: an experimental research]. (United States)

    Zhang, Yu-lian; Zhang, Lin-lin; Song, Wan-shan; Han, Wen-wen; Huang, Jian-hua; Zhou, Zhen


    To observe the regulatory effects of psoralen, oleanolic acid, and stilbene glucoside, three active components of psoralea fruit, glossy privet fruit and tuber fleeceflower root respectively, on Aβ25-35induced self-renewal and neuron-like differentiation of neural stem cells (NSCs). Embryonic NSCs werein vitro isolated and cultured from Kunming mice of 14-day pregnancy, and randomly divided into the control group, the Aβ25-35 group, the Aβ25-35 +psoralen group, the Aβ25-35 +oleanolic acid group, and the Aβ25-35 + stilbene glucoside group. The intervention concentration of Aβ25-35 was 25 µmol/L, and the intervention concentration of three active components of Chinese medicine was 10(-7)mol/L. The effect of three active components of Chinese medicine on the proliferation of NSCs was observed by counting method. The protein expression of Tubulin was observed by Western blot and immunofluorescence. The ratio of Tubulin+/DAPI was caculated. Results Compared with the control group, the sperical morphology of NSCs was destroyed in the Aβ25-35 group, the counting of NSCs, the expression of Tubulin protein, and the ratio of Tubulin /DAPI all decreased (P medicine treated groups (P <0. 01, P <0. 05). 25 µmol/L Aβ25-35 could inhibit self-renewal and neuron-like differentiating of NSCs. But psoralen, oleanolic acid, and stilbene glucoside could promote self-renewal of NSCs and neuron-like differentiation.

  1. Effects of 8-methoxypsoralen and ultraviolet light A on EGF receptor (HER-1) expression. (United States)

    Yang, X Y; Ronai, Z A; Santella, R M; Weinstein, I B


    Activation of psoralens by ultraviolet light irradiation at 308-400 nm (UVA) is used in the photochemical treatment of psoriasis. While the major effect of this activation is the formation of DNA adducts, it was recently demonstrated that psoralens can also bind to specific saturable high affinity cellular receptors, and that this is associated with inhibition of epidermal growth factor (EGF)-receptor binding. In view of these findings, we have examined whether 8-methoxy-psoralen (8-MOP) itself, or in combination with UVA, influences expression of the human EGF-receptor gene ("HER-1") in a human keratinocyte cell line. We have found that 8 MOP alone, and to a lesser extent UVA, induce a striking increase in cellular levels of HER-1 RNA. The combination of 8-MOP with UVA produces less induction of HER-1 RNA than that obtained with 8-MOP alone. We suggest, therefore, that this effect of 8-MOP is not due to DNA damage, but may reflect a separate effect of this compound on receptor-mediated signal transduction.

  2. The melanocortin 1 receptor (MC1R) inhibits the inflammatory response in Raw 264.7 cells and atopic dermatitis (AD) mouse model. (United States)

    Chen, Wei; Li, Jianping; Qu, Hai'e; Song, Zhou; Yang, Zhanqing; Huo, Jinlong; Jiang, Huaizhi; Huang, Qinghua; Huo, Meixia; Liu, Bo; Zhang, Qiaoling


    The alpha melanocyte stimulating hormone receptor (MC1R) is one of five G-protein coupled receptors belonging to the melanocortin subfamily, MC1R gene has been known to play a major role in regulating of fur color in mammals, and α-MSH and ACTH are endogenous nonselective agonists for MC1R. However, we found that MC1R was highly expressed in Raw 264.7 cells which were important inflammatory cells involved in the initiation of inflammatory responses. In addition, Cyclic AMP is not only a key molecule in the MC1R signal transduction pathway, but dampen innate immune-mediated responses. These intriguing biological results triggered the further conformation studies; it suggested that MC1R was very likely to be an important role in immunoregulation. In this study, we were to investigate the immunosuppressive effects of MC1R on inflammation in lipopolysaccharide (LPS) stimulated Raw 264.7 cells and LPS induced vivo 2-chloro-1,3,5-trinitrobenzene (TNCB)-induced atopic dermatitis (AD) model. The effects of the MC1R antagonist psoralen on pro-inflammatory cytokines and signaling pathways were analyzed by enzyme-linked immunosorbent assay, western blot, real-time fluorescence quantitative PCR and Histological analysis. Our results show a consistent and marked effect of high concentrations of MC1R antagonist psoralen increased the level of MC1R mRNA in Raw 264.7 cells by cumulative feedback regulation through preferential binding of MC1R. Moreover, as evidenced by inhibiting the LPS-induced TNF-α, IL-6 and enhancing the expression level of cyclic AMP protein in vitro. In vivo study it was also observed that psoralen promoted on histopathologic changes in the skin tissue of TNCB-induced AD mice. Taken together, our results suggest that MC1R decrease the inflammation in vitro and vivo, and might be a therapeutic signaling pathway to against inflammatory diseases.

  3. [Effects of P and K fertilizer on content of coumarin and yield of Glehnia littoralis]. (United States)

    Sun, Chuang-shu; Zheng, Kan; Li, Wei; Chen, Gui-lin; Yu, Rong; Yu, Jian-guo


    By a orthogonal experiment, the influence of different ratio of phosphorus and potassium fertilizers on imperatorin, isoimperatorin and psoralen contents and yield of Glehnia littoralis were studied. The results showed that root dry weight and the yield of G. littoralis increased when reasonably applied phosphorus fertilizer combined with potassium fertilizer within a certain range. And the influence of phosphorus fertilizer was greater than that of potassium fertilizer. The optimal value of root dry weight and yield achieved at both P2O5 360 kg x hm(-2), K2O 270 kg x hm(-2) and P2O5 360 kg x hm(-2), K2O 180 kg x hm(-2). The effects of different phosphorus and potassium treatments on the content of imperatorin, isoimperatorin and psoralen in G. littoralis were determined, which shows that the content increased with the moderate increase of phosphorus and potassium. And the effects of phosphorus fertilizer were more significantly. The isoimperatorin content achieved the largest value at P2O5 360 kg x hm(-2), K2O 270 kg x hm(-2), also a larger content of imperatorin and psoralen. The imperatorin content is the largest when applied P2O5 360 kg x hm(-2), K2O 180 kg x hm(-2), and the isoimperatorin content was higher as well. So that the treatment of P2O5 360 kg x hm(-2), K2O 180 kg x hm(-2) are suitable for promote to the agricultural production, which could improve the quality and yield of G. littoralis.

  4. Single Molecule Analysis of Laser Localized Interstrand Crosslinks

    Directory of Open Access Journals (Sweden)

    Jing eHuang


    Full Text Available DNA interstrand crosslinks (ICLs block unwinding of the double helix, and have always been regarded as major challenges to replication and transcription. Compounds that form these lesions are very toxic and are frequently used in cancer chemotherapy. We have developed two strategies, both based on immunofluorescence, for studying cellular responses to ICLs. The basis of each is psoralen, a photoactive (by long wave ultraviolet light, UVA DNA crosslinking agent, to which we have linked an antigen tag. In the one approach, we have taken advantage of DNA fiber and immunoquantum dot technologies for visualizing the encounter of replication forks with ICLs induced by exposure to UVA lamps. In the other, psoralen ICLs are introduced into nuclei in live cells in regions of interest (ROI defined by a UVA laser. The antigen tag can be displayed by conventional immunofluorescence, as can the recruitment and accumulation of DNA Damage Response (DDR proteins to the laser localized ICLs. However, substantial difference between the technologies creates considerable uncertainty as to whether conclusions from one approach are applicable to those of the other. In this report we have employed the fiber/quantum dot methodology to determine lesion density and spacing on individual DNA molecules carrying laser localized ICLs. We have performed the same measurements on DNA fibers with ICLs induced by exposure of psoralen to UVA lamps. Remarkably, we find little difference in the adduct distribution on fibers prepared from cells exposed to the different treatment protocols.. Furthermore, there is considerable similarity in patterns of replication in the vicinity of the ICLs introduced by the two techniques

  5. Psoriasis (chronic plaque). (United States)

    Naldi, Luigi; Rzany, Berthold


    Psoriasis affects 1-3% of the population, in some people causing changes to the nails and joints in addition to skin lesions. We conducted a systematic review and aimed to answer the following clinical questions: What are the effects of systemic drug treatments, topical drug treatments, and non-drug treatments (other than ultraviolet light) for chronic plaque psoriasis? What are the effects of ultraviolet light treatments for chronic plaque psoriasis? What are the effects of combined treatment with drugs plus ultraviolet light on chronic plaque psoriasis? What are the effects of combined systemic plus topical drug treatments for chronic plaque psoriasis? We searched: Medline, Embase, The Cochrane Library, and other important databases up to August 2007 (Clinical Evidence reviews are updated periodically; please check our website for the most up-to-date version of this review). We included harms alerts from relevant organisations such as the US Food and Drug Administration (FDA) and the UK Medicines and Healthcare products Regulatory Agency (MHRA). We found 122 systematic reviews, RCTs, or observational studies that met our inclusion criteria. We performed a GRADE evaluation of the quality of evidence for interventions. In this systematic review we present information relating to the effectiveness and safety of the following interventions: acupuncture, adding calcipotriol (topical) to psoralen plus ultraviolet light A or ultraviolet light B, adding oral retinoids to psoralen plus ultraviolet A (PUVA), alefacept, balneotherapy, ciclosporin, dithranol, T cell-targeted therapies, cytokine blocking agents, emollients (alone or plus ultraviolet light B), etanercept, fish oil supplementation, fumaric acid derivatives, Goeckerman treatment, heliotherapy, infliximab, Ingram regimen, keratolytics (salicylic acid, urea), leflunomide, methotrexate, oral pimecrolimus, phototherapy plus balneotherapy, psoralen plus ultraviolet A, psychotherapy, oral retinoids (alone or with

  6. Anthracene with near ultraviolet light inhibiting epidermal proliferation. (United States)

    Walter, J F; DeQuoy, P R


    Anthracene plus near ultraviolet (UV) light (UV-A, 320 to 400 nm) suppresses DNA synthesis and mitosis in mouse epidermis. Ultraviolet-A light or anthracene alone does not have any effect. There is no photoactivation of anthracene to enhance depression of DNA synthesis by either UV-B (290 to 320 nm) or UV-C (254 nm) light. While methoxsalen with UV-A light inhibits DNA synthesis, the phototoxic drugs chlorpromazine hydrochloride and demethylchlortetracycline do not. The combination of anthracene plus UV-A light may have therapeutic effectiveness for psoriasis with less potential for photocarcinogenesis than psoralens plus UV-A light.

  7. Psoralea corylifolia L. (Buguchi) - folklore to modern evidence: review. (United States)

    Chopra, Bhawna; Dhingra, Ashwani Kumar; Dhar, Kanaya Lal


    Psoralea corylifolia is an important medicinal plant which is used in several traditional medicines to cure various diseases. The plant extracts have been reported to possess antibacterial, antitumor, antioxidant, anti-inflammatory, antifungal and immunomodulatory activity. A wide range of chemical compounds including psoralen, isopsoralen, bakuchiol, psoralidin, bakuchalcone, bavachinin, flavones, volatile oils, lipids etc. are found in different parts of the plant. The present review is therefore, an effort to give a detailed survey of the literature on its botany, phytochemistry and ethnopharmacology along with special emphasis given on pharmacological activities of plant P. corylifolia. © 2013.

  8. Systemic therapies for psoriasis: understanding current and newly emerging therapies. (United States)

    Sobell, Jeffrey M; Hallas, Stephen J


    The treatment of moderate to severe psoriasis is a rapidly expanding area. Recent insights into the pathogenesis of this disease as a T-cell mediated process has led to a greater understanding of the mechanisms of action of conventional FDA-approved systemic therapies such as methotrexate, cyclosporine, acitretin, and psoralen with ultraviolet A phototherapy. It has also led to the development of rationally targeted therapies against key components of the immune process critical in the generation of the psoriatic plaque. Safety and efficacy data from clinical studies of 4 biologic agents furthest along in their development are reviewed. These results are promising, adding to the armamentarium for treating this disease.

  9. Integral parametrization of the Kinetics of Crosslink production in plasmid DNA as a function of 8-methoxypsoralen concentration

    Energy Technology Data Exchange (ETDEWEB)

    Vidania, R. de; Paramio, J. M.; Bauluz, C.


    In this paper we present results of crosslink production in pBR322 DNA along a wide range of 8-methoxypsoralen (8-MOP) concentration. Experimental data were obtained as DNA renaturation percentages, from the shift in hyperchromicity after a temperature-dependent denaturation-renaturation process. the experimental results showed a three-stage profile when represented as a function of the natural logarithms of 8-MOP concentration. an integral parametrization which allows a simultaneous fit of the three observed stages is presented here. the theoretical values of crosslink production determined from the fit are useful to asses the genotoxicity of psoralen-induced crosslinks in plasmid DNA. (Author) 24 refs.

  10. Current Approach to Phototherapy Applications in Dermatology Practice: Part 2. Phototherapy in Vitiligo, Mycosis Fungoides, Atopic Dermatitis, Sclerosing Diseases and Psoriasis

    Directory of Open Access Journals (Sweden)

    Dilek Seçkin Gençosmanoğlu


    Full Text Available Many skin diseases can be treated with phototherapy, however, the method choice depends on the disease’s characteristics. First-line phototherapy for generalized plaque psoriasis is narrowband ultraviolet B whereas it is psoralen and ultraviolet A for plaque stage mycosis fungoides and ultraviolet A1 for severe localized scleroderma. In this review, efficacy of different phototherapy methods in the most common diseases treated with phototherapy such as vitiligo, mycosis fungoides, atopic dermatitis, sclerosing diseases and psoriasis will be discussed in light of current guidelines.

  11. Vitiligo following type II lepra reaction. (United States)

    Pavithran, K


    A middle-aged male with lepromatous leprosy developed bouts of skin lesions of depigmented macules and patches of vitiligo, just following attacks of type II lepra reaction each time. In view of the present concept of autoimmunity playing a role in the pathogenesis of vitiligo as well as lepra reaction, their association in our patient appears to be more than fortuious. The depigmented macules persisted even after regression of skin lesions of leprosy following chemotherapy. The vitiligo macules responded partially to topical and systemic psoralen therapy.

  12. Synthesis of reactive nucleic acid analogues and their application for the study of structure and functions of biopolymers

    Energy Technology Data Exchange (ETDEWEB)

    Kanevskii, Igor' E; Kuznetsova, Svetlana A [Department of Chemistry, M.V. Lomonosov Moscow State University, Moscow (Russian Federation)


    Data on the synthesis of reactive derivatives of nucleic acid analogues and their application for the study of structure and functions of biopolymers are generalised. The main types of such analogues including photoactivated reagents containing azidoaryl, halogeno, and thiol groups, psoralen and its derivatives, platinum-based reagents, and nucleic acid analogues containing substituted pyrophosphate or acyl phosphate internucleotide groups are presented. The mechanisms of interaction of these compounds with proteins and nucleic acids are considered. The prospects for the in vivo application of reactive nucleic acids in various systems are discussed. The bibliography includes 76 references.

  13. Association between the synthetic vascular stent used for abdominal aortic aneurysm and generalized granuloma annulare

    Directory of Open Access Journals (Sweden)

    Han-Chi Tseng


    Full Text Available The etiology of granuloma annulare (GA remains unknown, although several eliciting factors have been proposed. We herein present the case of an 81-year-old man who developed generalized erythematous annular plaques 6 months after engraftment of a vascular stent for abdominal aneurysm repair. Based on the diagnosis of generalized GA and the patient's age, we treated him with psoralens plus ultraviolet A therapy. The treatment response was good. This is the first report showing the association between vascular stent and generalized GA.

  14. Phytochemical analysis of Ferulogo Bernardii Tomk & M.Pimen

    Directory of Open Access Journals (Sweden)

    Khalighi-Sigaroodi F.


    Full Text Available From the hexane extract of the aerial parts of Ferulago Bernardii (Apiaceae four coumarins, namely prantschimgin 1, oxypeucedanin 2, psoralen 3 and umbelliferone 4; β-sitosterol 5; and nonacosane 6 were isolated by Column Chromatography (CC, Preparative Thin Layer Chromatography (PTLC and crystallization. The structures were elucidated by melting point, UV, IR, MS, 1H and 13C-NMR spectra. The presence of compounds 1, 2, 3 and 5 in some others Ferulago species could be used as chemotaxonomic marker in genus Ferulago. This is the first report on phytochemical analysis of Ferulago Bernardii Tomk. & M. Pimen.

  15. Encapsulation of porcine pancreatic islets within an immunoprotective capsule comprising methacrylated glycol chitosan and alginate. (United States)

    Hillberg, Anna Louise; Oudshoorn, Matthew; Lam, Janice B B; Kathirgamanathan, Kalyani


    Encapsulation of cells in biocompatible polymer matrices represents a powerful tool for cell-based therapies and therapeutic delivery systems. This technology has successfully been used to deliver pancreatic islets to humans for the treatment of Type 1 diabetes. However, the clinical impact of this technology may be improved by reducing the inflammatory response brought on after implantation of capsules in vivo. Within this study a biocompatible polymeric delivery system combining alginate and photo-crosslinked methacrylated glycol chitosan (MGC) was developed. This approach involved encapsulating cells in calcium-alginate beads, coating with MGC and photo-polymerizing using UVA in the presence of photo-initiator (VA-086), resulting in the formation of capsules ∼600 µm in size. Crosslinking of the MGC outer wall allowed control over capsule swelling and improved the capsules overall properties. Capsule characterization demonstrated the stabilizing influence of polymerization and fluorescence imaging showed that the distribution of glycol chitosan is dependent on molecular weight. Good islet viability and insulin release was demonstrated in vitro over the course of a month, and in vivo transplantation of the capsules demonstrated good biocompatibility, particularly when compared with standard alginate/poly-l-ornithine/alginate capsules. © 2014 Wiley Periodicals, Inc.

  16. Fluorescence spectroscopy of collagen crosslinking: non-invasive and in situ evaluation of corneal stiffness (United States)

    Franco, Walfre; Ortega-Martinez, Antonio; Zhu, Hong; Wang, Ruisheng; Kochevar, Irene E.


    Collagen is a long fibrous structural protein that imparts mechanical support, strength and elasticity to many tissues. The state of the tissue mechanical environment is related to tissue physiology, disease and function. In the cornea, the collagen network is responsible for its shape and clarity; disruption of this network results in degradation of visual acuity, for example in the keratoconus eye disease. The objective of the present study is to investigate the feasibility of using the endogenous fluorescence of collagen crosslinks to evaluate variations in the mechanical state of tissue, in particular, the stiffness of cornea in response to different degrees of photo-crosslinking or RGX treatment—a novel keratoconus treatment. After removing the epithelium, rabbit corneas were stained with Rose Bengal and then irradiated with a 532 nm solid-state laser. Analysis of the excitation spectra obtained by fluorescence spectroscopy shows a correlation between the fluorescence intensity at 370/460 nm excitation/emission wavelengths and the mechanical properties. In principle, it may be feasible to use the endogenous fluorescence of collagen crosslinks to evaluate the mechanical stiffness of cornea non-invasively and in situ.

  17. Synthesis and properties of novel photosensitive poly(arylene ether sulfone) containing chalcone moiety in the main chain

    Energy Technology Data Exchange (ETDEWEB)

    Wen Pushan [Key Laboratory of Catalysis and Materials Science of the State Ethnic Affairs Commission and Ministry of Education, Hubei Province, South-Central University for Nationalities, Wuhan 430074 (China); Wang Lei [Department of Materials Physics and Chemistry, School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083 (China); Zhang Aiqing [Key Laboratory of Catalysis and Materials Science of the State Ethnic Affairs Commission and Ministry of Education, Hubei Province, South-Central University for Nationalities, Wuhan 430074 (China); Li Xiangdan, E-mail: [Key Laboratory of Catalysis and Materials Science of the State Ethnic Affairs Commission and Ministry of Education, Hubei Province, South-Central University for Nationalities, Wuhan 430074 (China); Lee, Myong-Hoon, E-mail: [Department of Polymer/Nano Science and Technology, Chonbuk National University, Jeonju 561-756 (Korea, Republic of)


    Research highlights: {yields} A series of novel photosensitive poly(arylene ether sulfone)s (PAESs) containing chalcone moiety in the main chain have been successfully synthesized. {yields} The photo-crosslinking of polymer film was carried out under UV irradiation without photoinitiator. {yields} The resulting polymers showed good thermal stability and excellent chemical stability after crosslinking. - Abstract: A new series of photosensitive poly(arylene ether sulfone)s containing chalcone moiety in the main chain were synthesized from 4,4'-dihydroxychalcone (4DHC), 4,4'-difluorodiphenylsulfone (DFDPS) and bisphenol A (BPA). This series of polymers were characterized by {sup 1}H NMR, FT-IR, UV spectroscopy, thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC). The polymers were stable up to 400 deg. C, which indicates that the polymers possess good thermal properties. The polymers were found to be soluble in polar solvents and chlorinated solvents. However, the polymers were insoluble in hydrocarbons and in hydroxyl group-containing solvents. After the irradiation of UV light, the thin polymer film was crosslinked to give an insoluble film in the absence of a photoinitiator or sensitizers. The rate of photocrosslinking was also examined and discussed.

  18. Flexible polymer waveguides for light-activated therapy (Conference Presentation) (United States)

    Kim, Moonseok; Kwok, Sheldon J. J.; Lin, Harvey H.; Lee, Dong Hee; Yun, Seok Hyun


    Conventional light-activated therapies, such as photodynamic therapy (PDT), photochemical tissue bonding (PTB), collagen crosslinking (CXL), low-level light therapy (LLLT), and antimicrobial therapy utilize external light sources and light propagation through free space, limiting treatment to accessible and superficial areas of the body. Recent progress has been made in developing biocompatible polymer waveguides to enhance light delivery to deep tissues. To further expand clinical utility, waveguides should be flexible and tough enough to enable use in anatomically difficult-to-reach regions, while having the requisite optical properties to achieve uniform and efficient illumination of the target area. Here, we present a new class of flexible polymer waveguides optimized for uniform light extraction into tissues. Our slab waveguides comprise two designs: first, a flexible polydimethylsiloxane (PDMS) based elastomer for CXL, and second, a tough polyacrylamide and alginate hydrogel for large-area phototherapies. Our waveguides are optically transparent in the visible wavelengths (400-750 nm) and a multimode fiber is used to couple light into the waveguide. We characterized the light propagation through the waveguides and light extraction into tissue, and validated our results with optical simulation. By changing the thickness and scattering properties, uniform light extraction through the length of the waveguide could be achieved. We demonstrate proof-of-concept scleral photo-crosslinking of an ex vivo porcine eyeball for prevention of myopia.

  19. Development of Electrically Conductive Double-Network Hydrogels via One-Step Facile Strategy for Cardiac Tissue Engineering. (United States)

    Yang, Boguang; Yao, Fanglian; Hao, Tong; Fang, Wancai; Ye, Lei; Zhang, Yabin; Wang, Yan; Li, Junjie; Wang, Changyong


    Cardiac tissue engineering is an effective method to treat the myocardial infarction. However, the lack of electrical conductivity of biomaterials limits their applications. In this work, a homogeneous electronically conductive double network (HEDN) hydrogel via one-step facile strategy is developed, consisting of a rigid/hydrophobic/conductive network of chemical crosslinked poly(thiophene-3-acetic acid) (PTAA) and a flexible/hydrophilic/biocompatible network of photo-crosslinking methacrylated aminated gelatin (MAAG). Results suggest that the swelling, mechanical, and conductive properties of HEDN hydrogel can be modulated via adjusting the ratio of PTAA network to MAAG network. HEDN hydrogel has Young's moduli ranging from 22.7 to 493.1 kPa, and its conductivity (≈10(-4) S cm(-1)) falls in the range of reported conductivities for native myocardium tissue. To assess their biological activity, the brown adipose-derived stem cells (BADSCs) are seeded on the surface of HEDN hydrogel with or without electrical stimulation. Our data show that the HEDN hydrogel can support the survival and proliferation of BADSCs, and that it can improve the cardiac differentiation efficiency of BADSCs and upregulate the expression of connexin 43. Moreover, electrical stimulation can further improve this effect. Overall, it is concluded that the HEDN hydrogel may represent an ideal scaffold for cardiac tissue engineering. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Protein 'smart' micro/nano-biooptics via femtosecond laser direct writing (United States)

    Sun, Yun-Lu; Dong, Wen-Fei; Sun, Hong-Bo


    Protein-hydrogel-based free-form three-dimensional (3D) micro/nano-elements with ``smart'' stimuli-responsiveness fabricated via femtosecond laser direct writing (FsLDW) have attracted increasing efforts for their wide utilities in many fields, such as bio-micro/nano-machine [1], biophotonics [2, 3], etc. All processes of this protein-FsLDW are carried out in proteins' aqueous solutions, as a result of which the approach is noncontact, maskless and biocompatible. So, we applied the promising FsLDW approach, which, in the construction of protein-based 3D optical microdevices, for example, microlenses. In order to meet the requirements of optical applications, FsLDW system and processing parameters were carefully optimized to achieve high-quality surface and 3D morphology. The photo-crosslinked protein microhydrogels showed a rapid and reversible swell-to-shrink behavior once stimulated by chemical signals, by which the protein microdevices can be dynamically tuned. Because of using protein molecules as "building blocks", protein-based microelements were demonstrated of good biocompatibility. Based on the valuable and unique merits, protein-based optical micro/nano-elements have great potential for micro/nano-bio-optics, bionics, and novel biomedical applications, etc.

  1. Synthesis and characterization of hybrid cured poly(ether-urethaneacrylate/titania microcomposites formed from tetraalkoxytitanate precursor

    Directory of Open Access Journals (Sweden)


    Full Text Available Hybrid poly(ether-urethaneacrylate (PEUA/titania (TiO2 microcomposites were prepared using a novel method that includes a swelling of different photo-crosslinked PEUA networks in concentrated tetraisopropoxytitanate (Ti(OiC3H74 or TIPT precursor solution in organic media followed by the hydrolysis of covalently bonded polyalkoxytitanate ([–OTi(OC3H72–]n chains and their aggregation to amorphous micro- and nano-scale sized TiO2 particles. A formation of polymer/titania hybrids was confirmed by complex investigations of the hybrids using infrared (IR spectroscopy, small angle X-ray scattering (SAXS analysis, scanning electron microscopy (SEM and gravimetry. The dependence of titania phase formation behavior versus functionality of the poly(ether-urethaneacrylate network was discussed. The presence of reactive groups in the organic network promotes the formation of surface-bonded ball-shaped type TiO2 inclusions as well as provides transparency to the hybrid film samples. The results obtained in this work can be applied for the development of polymer/TiO2 composite materials for multipurpose optical application and advanced sealants.

  2. Effects of permeability and living space on cell fate and neo-tissue development in hydrogel-based scaffolds: a study with cartilaginous model. (United States)

    Fan, Changjiang; Wang, Dong-An


    One bottleneck in tissue regeneration with hydrogel scaffolds is the limited understanding of the crucial factors for controlling hydrogel's physical microenvironments to regulate cell fate. Here, the effects of permeability and living space of hydrogels on encapsulated cells' behavior were evaluated, respectively. Three model hydrogel-based constructs are fabricated by using photo-crosslinkable hyaluronic acid as precursor and chondrocytes as model cell type. The better permeable hydrogels facilitate better cell viability and rapid proliferation, which lead to increased production of extracellular matrix (ECM), e.g. collagen, glycosaminoglycan. By prolonged culture, nano-sized hydrogel networks inhibit neo-tissue development, and the presence of macro-porous living spaces significantly enhance ECM deposition via forming larger cell clusters and eventually induce formation of scaffold-free neo-tissue islets. The results of this work demonstrate that the manipulation and optimization of hydrogel microenvironments, namely permeability and living space, are crucial to direct cell fate and neo-tissue formation. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. A comparative study of zwitterionic ligands-mediated mineralization and the potential of mineralized zwitterionic matrices for bone tissue engineering. (United States)

    Liu, Pingsheng; Emmons, Erin; Song, Jie


    Cationic and anionic residues of the extracellular matrices (ECM) of bone play synergistic roles in recruiting precursor ions and templating the nucleation, growth and crystalline transformations of calcium apatite in natural biomineralization. We previously reported that zwitterionic sulfobetaine ligands can template extensive 3-dimensional (3-D) hydroxyapaptite (HA)-mineralization of photo-crosslinked polymethacrylatehydrogels. Here, we compared the potency of two other major zwitterionic ligands, phosphobetaine and carboxybetaine, with that of the sulfobetaine in mediating 3-D mineralization using the crosslinked polymethacrylate hydrogel platform. We confirmed that all three zwitterionic hydrogels were able to effectively template 3-D mineralization, supporting the general ability of zwitterions to mediate templated mineralization. Among them, however, sulfobetaine and phosphobetaine hydrogels templated denser 3-D mineralizationthan the carboxybetaine hydrogel, likely due to their higher free water fractions and better maintenance of zwitterionic nature throughout the pH-changes during the in vitro mineralization process. We further demonstrated that the extensively mineralized zwitterionic hydrogels could be exploited for efficient retention (e.g. 99% retention after 24-h incubation in PBS) of osteogenic growth factor recombinant bone morphogenetic protein-2 (rhBMP-2) and subsequent sustained local release with retained bioactivity. Combined with the excellent cytocompatibility of all three zwitterionic hydrogels and the significantly improved cell adhesive properties of their mineralized matrices, these materials could find promising applications in bone tissue engineering.

  4. Post-transcriptional m6A editing of HIV-1 mRNAs enhances viral gene expression (United States)

    Kennedy, Edward M.; Bogerd, Hal P.; Kornepati, Anand V. R.; Kang, Dong; Ghoshal, Delta; Marshall, Joy B.; Poling, Brigid C.; Tsai, Kevin; Gokhale, Nandan S.; Horner, Stacy M.; Cullen, Bryan R.


    Summary Covalent addition of a methyl group to the adenosine N6 (m6A) is an evolutionarily conserved and common RNA modification that is thought to modulate several aspects of RNA metabolism. While the presence of multiple m6A editing sites on diverse viral RNAs was reported starting almost 40 years ago, how m6A editing affects virus replication has remained unclear. Here, we used photo-crosslinking-assisted m6A sequencing techniques to precisely map several m6A editing sites on the HIV-1 genome and report that they cluster in the HIV-1 3’ untranslated region (3'UTR). Viral 3'UTR m6A sites or analogous cellular m6A sites strongly enhanced mRNA expression in cis by recruiting the cellular YTHDF m6A “reader” proteins. Reducing YTHDF expression inhibited, while YTHDF overexpression enhanced, HIV-1 protein and RNA expression, and virus replication in CD4+ T cells. These data identify m6A editing, and the resultant recruitment of YTHDF proteins, as major positive regulators of HIV-1 mRNA expression. PMID:27117054

  5. Effect of rutin on virus inactivation by AMT in combination with ultraviolet-A irradiation in platelet concentrates

    Energy Technology Data Exchange (ETDEWEB)

    Yamada, Yoshiko; Abe, Hideki; Ikebuchi, Kenji; Sekiguchi, Sadayoshi [Hokkaido Red Cross Blood Center, Sapporo (Japan)


    Treatment with psoralens and ultraviolet-A (UVA) irradiation have been found to be effective for virus sterilization of platelet concentrates (PCs). We report here a virus inactivation method using a combination of psoralen derivative 4'-aminomethyl-4,5', 8-trimethylpsoralen (AMT) and UVA irradiation (AMT/UVA). Further, we also investigated the effect of rutin, a radical scavenger, on the inactivation of vesicular stomatitis virus (VSV) as a model virus administered in PCs and platelet functions were investigated. Spiked VSV (about 5log{sub 10}) in PCs was inactivated by a combination of AMT (50 {mu}g/ml) and 5.2 J/cm{sup 2} UVA irradiation in the absence of rutin. To obtain equivalent levels of VSV kill in the presence of 0.35 mM rutin, treatment with 13.0 J/cm{sup 2} of UVA irradiation with AMT was performed. When PCs were treated under each condition in which 5log{sub 10} VSV was inactivated by AMT/UVA with or without rutin, platelet aggregation function was maintained for more than 80% of untreated platelets. These findings indicate that the presence of rutin during AMT/UVA treatment conferred no beneficial effect. In addition, overnight storage of PCs with AMT induced 40% loss of platelet aggregation in response to 10{mu}M ADP. The findings suggest that UVA irradiation is required immediately after the addition of AMT. (author)

  6. Study of efficacy of bath PUVA therapy in the treatment of generalized plaque type psoriasis

    Directory of Open Access Journals (Sweden)

    Azadeh S


    Full Text Available Psoriasis is a chronic, inflammatory scaling disorder of the skin. Different patterns of psoriasis exist including plaque type, erythrodemic, pustular, palmoplantar and guttate. The most commonly involved sites are the elbows, knees, lumbosacral area and scalp. PUVA (Psoriasis Plus UVA therapy [administration of oral psoralen followed by exposure to UVA (320 to 440 nm] is widely used to treat severe psoriasis. Oral PUVA produces some adverse effects that may limit its applicability in a number of patients. The carcinogenic potential limits its use in patients with psoriasis who probably receive other carcinogenic treatments. Oral PUVA may induce complications such as nausea, vomiting and headache. In light of these problems Bath PUVA therapy is an important alternative to oral PUVA therapy. Bath PUVA is a kind of photochemotherapy in which UVA radiation after administration of topical psoralen in a warm water bath is used. We treated 30 patients with generalized plaque type psoriasis with 8-Mop Bath PUVA in Razi hospital. Bath PUVA cleared psoriasis more rapidly than oral PUVA and required fewer treatments (mean number of sessions: (17.6±2.1 and lower cumulative UVA dose. (49.2±15.4 J/cm². 83.3 percent of our patients showed complete response to treatment and 13.4 percent showed good response.

  7. Nematicidal activity of furanocoumarins from parsley against Meloidogyne spp. (United States)

    Caboni, Pierluigi; Saba, Marco; Oplos, Chrisostomos; Aissani, Nadhem; Maxia, Andrea; Menkissoglu-Spiroudi, Urania; Casu, Laura; Ntalli, Nikoletta


    This report describes activity against Meloidogyne spp. and chemical characterisation of the essential oil and methanol extract of Petroselinum crispum aerial parts. The study was based on the hypothesis that P. crispum could be used as an intercrop and soil amendment in tomato culture for nematode control. The methanol extract and the essential oil exhibited significant nematicidal activity against M. incognita, M. hapla and M. arenaria, the first being the most sensitive species, with EC50 /72 h values of 140 ± 15 and 795 ± 125 mg L(-1) for the extract and oil respectively. The most abundant furanocoumarin compounds in the methanolic extract were xanthotoxin, psoralen, bergapten and oxypeucedanin; levels ranged from 1.77 to 46.04 mg kg(-1) wet weight. The EC50 /24 h values of xanthotoxol, psoralen and xanthotoxin against M. incognita were 68 ± 33, 147 ± 88 and 200 ± 21 mg L(-1) respectively. The addition of fresh parsley paste to soil reduced the number of M. incognita females and plant galls on tomato roots; EC50 values were 24.79 and 28.07 mg g(-1) respectively. Moreover, parsley paste enhanced tomato growth in a dose-response manner. Parsley exhibits promising nematicidal activity as an organic amendment and as a source of nematotoxic furanocoumarins. © 2014 Society of Chemical Industry.

  8. Uji Efektivitas Jeruk Nipis (Citrus aurantifolia dalam Mempercepat Laju Disinfeksi Bakteri Escheria Coli pada Proses Solar Water Disinfection

    Directory of Open Access Journals (Sweden)

    Saprian Saprian


    Full Text Available Solar Water Disinfection (SODIS memerlukan waktu rata-rata berkisar antara 5-6 jam untuk dapat membunuh bakteri 3-4 Log pada air berkapasitas maksimal 2 liter atau 1 jam dengan suhu di atas 50°C. Penelitian terbaru menunjukkan bahwa dengan mengkombinasikan SODIS dengan psoralens, zat aktif seperti yang terdapat pada jeruk Lemon (Citrus Limon dapat menginaktivasi bakteri hingga 5,6 Log reduksi dalam rentang waktu 30 menit pada suhu 29°C. Mengingat faktor ketersediaan dan harga jeruk lemon di Indonesia maka pada penelitian ini dilakukan pengamatan dengan menggunakan jeruk nipis (Citrus aurantifolia. Penelitian ini dilakukan untuk mengetahui seberapa besar potensi jeruk nipis untuk dipergunakan dalam mempercepat laju disinfeksi pada proses SODIS. Adapun faktor-faktor yang akan dipelajari terkait dengan hubungan antara variasi dosis jeruk nipis, perubahan pH dan suhu air terhadap efisiensi inaktivasi bakteri yang mampu dihasilkan. Variasi jeruk nipis yang digunakan adalah 0%, 2%, 4% dan 6% dengan variasi waktu 0 menit, 30 menit, 60 menit dan 120 menit. Hasil penelitian menunjukkan bahwa perlakuan SODIS dengan menggunakan jeruk nipis mampu menginaktivasi E. coli sebesar 5.4 Log dengan waktu 1 jam pada suhu 32°C. Hal ini menunjukkan bahwa Jeruk nipis berpotensi untuk digunakan dalam mempercepat laju disinfeksi E. coli pada air minum. Kata Kunci : SODIS, E.coli, Psoralens, Jeruk Nipis

  9. Instrumentation and action spectra in light-associated diseases

    Energy Technology Data Exchange (ETDEWEB)

    Cripps, D.J.


    Instrumentation for studying action spectra in controls and various light-associated diseases is described. This study summarizes tests performed with a prism grating monochromator during the last 10 yr. There were 68 photodermatoses studied: xeroderma pigmentosum (XP) (1), lupus erythematosus (LE) (12), polymorphous light eruption (PLE) (23), solar urticaria (4), actinic reticuloid (2), halogenated salicylanilide photosensitivity and persistent light reactors (11), psoralen photosensitivity (6), and porphyria (9). A normal minimal erythema dose in the UVB (below 320 nm) was generally observed in polymorphous light eruption and lupus erythematosus. The most exquisite photosensitivity for delayed erythema was observed in actinic reticuloid, which in one case was 25-35 times more sensitive in the UVB range which was also observed but to a lesser extent in XP and in persistent light reactors. Persistence of erythema and edema at test sites was observed in XP, PLE, LE, and actinic reticuloid. A delay in development of erythema reaching a maximum at 72 hr was observed in XP and psoralen phototoxicity. Maximum photosensitivity occurred in solar urticaria. Three patients had peak sensitivity in the range of 310-313 nm and the 4th at 460 nm. Photosensitivity in the visible range was detected in 2 patients with solar urticaria, one with actinic reticuloid, and confirmed in 9 patients with porphyria (405 nm). Photosensitivity in the UVA (above 320 nm) occurred to some degree in all groups.

  10. New Limonoids from Hortia oreadica and Unexpected Coumarin from H. superba Using Chromatography over Cleaning Sephadex with Sodium Hypochlorite

    Directory of Open Access Journals (Sweden)

    Vanessa G.P. Severino


    Full Text Available Previous investigations of H. oreadica reported the presence of a wide spectrum of complex limonoids and dihydrocinnamic acids. Our interest in the Rutaceae motivated a reinvestigation of H. oreadica, H. brasiliana and H. superba searching for other secondary metabolites present in substantial amounts for taxonomic analysis. In a continuation of the investigation of the H. oreadica, three new limonoids have now been isolated 9α-hydroxyhortiolide A, 11β-hydroxyhortiolide C and 1(S*-acetoxy-7(R*-hydroxy-7-deoxoinchangin. All the isolated compounds from the Hortia species reinforce its position in the Rutaceae. With regard to limonoids the genus produces highly specialized compounds, whose structural variations do not occur in any other member of the Rutaceae, thus, it is evident from limonoid data that Hortia takes an isolated position within the family. In addition, H. superba afforded the unexpected coumarin 5-chloro-8-methoxy-psoralen, which may not be a genuine natural product. Solid-state cross-polarisation/magic-angle-spinning 13C nuclear magnetic resonance, X-Ray fluorescence and Field-emission gun scanning electron microscopy experiments show that the Sephadex LH-20 was modified after treatment with NaOCl, suggesting that when xanthotoxin (8-methoxy-psoralen was extracted from cleaning of the gel column, chlorination of the aromatic system occurred.

  11. Chemotherapeutic treatment is associated with Notch1 induction in cutaneous T-cell lymphoma

    DEFF Research Database (Denmark)

    Kamstrup, Maria R; Biskup, Edyta; Manfè, Valentina


    The Notch pathway is important for survival of cutaneous T-cell lymphoma (CTCL) cells. We investigated the effect of chemotherapy (doxorubicin, etoposide, and gemcitabine) and radiation modalities on Notch signaling in CTCL cell lines. Chemotherapy induced Notch1 expression at the mRNA and protein...... level in MyLa2000 and Hut78. Upregulation of well-established Notch targets supported the functional activity of Notch1. Transfection of Notch1 siRNA into MyLa2000 cells was not able to suppress the effects of chemotherapy on Notch1 activation significantly. Notch1 knockdown in combination...... with doxorubicin, etoposide, or gemcitabine compared to chemotherapy alone decreased cell viability by 12, 20, and 26%, respectively (p MyLa2000 but not SeAx) and psoralen plus UVA (PUVA) (in MyLa2000, Hut78, and SeAx) increased the expression of Notch1 family members. Our results...

  12. Photodynamic decontamination of blood for transfusion (United States)

    Ben-Hur, Ehud; Margolis-Nunno, H.; Gottlieb, P.; Lustigman, S.; Horowitz, Bernard


    Currently transfused cellular components of blood are not available in a sterile form and carry a small risk of transmitting viral and parasite diseases. Using phthalocyanines and red light, lipid enveloped viruses, e.g., HIV-1, can be inactivated in red blood cell concentrates (RBCC). Under conditions leading to virus sterilization the blood borne parasites Trypanosoma cruzi (Chagas disease) and Plasmodium falciparum (malaria) could be eliminated to undetectable levels (> 4 log10 kill). RBC damage during treatment could be avoided by increasing the light fluence rate to 80 mW/cm2, and by including the free radical scavenger glutathione and the vitamin E derivative Trolox during light exposure. Similar sterilization of platelet concentrates was achieved with the psoralen derivative AMT and UVA light. Platelet damage due to PUVA treatment was avoided by including the plant flavonoid rutin during irradiation. It is concluded that elimination of the risk of transmitting pathogens during blood transfusion is feasible with photochemical treatments.

  13. Recalcitrant granuloma faciale with 14-year follow-up

    Directory of Open Access Journals (Sweden)

    Keiko Shiba


    Full Text Available Granuloma faciale (GF is an uncommon inflammatory dermatosis that is characterized by one or several asymptomatic brown red nodules or plaques usually occurring on the face. GF is known to be extremely resistant to treatment. Various treatments have been attempted, including topical or intralesional or systemic corticosteroids, dapsone, antimalarials, colchicine, topical tacrolimus, and topical psoralen; however, none of these have been found to be reliably efficacious. We tried numerous combinations and rotations of therapeutic modalities for GF over a long time. We described a case of GF in which the eruptions were carefully observed over the course of 14 years before finally resolving. Our experience suggests that continuous and careful follow-up, and trials of combinations and rotations of therapeutic modalities are essential for treating GF.

  14. Singlet oxygen reactivity in water-rich solvent mixtures

    Directory of Open Access Journals (Sweden)

    Cristina Sousa


    Full Text Available The 3-methylindole (3MI oxygenation sensitized by psoralen (PSO has been investigated in 100%, 20% and 5% O2-saturated water/dioxane (H2O/Dx mixtures. The lowering of the ¹O2* chemical rate when water (k chem∆3MI = 1.4 × 109 M-1 s-1 is replaced by deuterated water (k chem∆3MI = 1.9 × 108 M-1 s-1 suggests that hydrogen abstraction is involved in the rate determining step. A high dependence of the chemical rate constant on water concentration in H2O/Dx mixtures was found showing that water molecules are absolutely essential for the success of the 3MI substrate oxidation by ¹O2* in water-rich solvent mixtures.

  15. [Studies on coumarins from fruit of Cnidium monnieri and their cytotoxic activities]. (United States)

    Duan, Xu-hong; Zhang, Yu-zhuo; He, Pei; Ma, Zong-min; Pei, Lin


    This study is to study is to investigate the coumarins from Fruit of Cnidium monnieri and their cytotoxic activities. The constituents were separated by column chromatography, and their structures were elucidated by spectroscopic data analyses. The isolated compounds were evaluated for their cytoxic activities by MTT method. Eleven compounds were isolated and identified as osthole (1), bergaptan (2), xanthotoxol (3), xanthotoxin (4), imperatorin (5), isopimpinellin (6), osthenol (7), psoralen (8), 5,7-dimethoxycoumarin (9), oxypeucedaninhydrate (10), and swietenocoumarin F (11). Compounds 7, 9-11 were isolated from the Cnidium genus for the first time. Compounds 1,5,10 and 11 showed significant cytotoxic activities against L1210 cell lines at a concentration of 1 x 10(-5) mol x L(-1) with inhibitory rates of were 70.13, 63.10, 55.77, and 75.08% respectively.


    Directory of Open Access Journals (Sweden)

    Yogeesh Hosahalli Rajaiah


    Full Text Available Polymprhouos light eruption is the most common idiopathic photodermatosis. It is a sun induced cutaneous reaction characterized by onset itchy erathematous papules, plaques, vesicles or erythema multiforme type of lesions after brief exposure to sunlight. Sun-exposed areas of the body or rarely the partially covered areas are commonly involved. PLE is more common in temperate climates than in tropics. It begins usually at the onset of summer and moderates as the summer progresses. In most patients it usually runs a benign course. Diagnosis is mainly on clinical grounds. Therapy involves avoidance of sun-exposure and use of sunscreens. Cases not responding to simple measures require PUVA (Psoralen and Ultraviolet A or UVB (ultraviolet B therapy. Other alternative suggested therapies with variable success include oral hydroxychloroquine, beta-carotene, thalidomide and nicotinamide.

  17. Human embryonic stem cells have enhanced repair of multiple forms of DNA damage

    DEFF Research Database (Denmark)

    Maynard, Scott; Swistowska, Anna Maria; Lee, Jae Wan


    Embryonic stem cells need to maintain genomic integrity so that they can retain the ability to differentiate into multiple cell types without propagating DNA errors. Previous studies have suggested that mechanisms of genome surveillance, including DNA repair, are superior in mouse embryonic stem...... cells compared with various differentiated murine cells. Using single-cell gel electrophoresis (comet assay) we found that human embryonic stem cells (BG01, I6) have more efficient repair of different types of DNA damage (generated from H2O2, UV-C, ionizing radiation, or psoralen) than human primary...... fibroblasts (WI-38, hs27) and, with the exception of UV-C damage, HeLa cells. Microarray gene expression analysis showed that mRNA levels of several DNA repair genes are elevated in human embryonic stem cells compared with their differentiated forms (embryoid bodies). These data suggest that genomic...

  18. Characterization of products from the pyrolysis of rapeseed oil cake. (United States)

    Ucar, Suat; Ozkan, Ahmet R


    The main aim of this study was to investigate the composition of products from the pyrolysis of rapeseed oil cake in a fixed bed reactor at 400, 450, 500, 700 and 900 degrees C. The gas products mainly consisted of CO(2), CO, CH(4) and H(2)S at 500 degrees C. Empirical formula of bio-oil from the pyrolysis of rapeseed oil cake was CH(1.59)O(0.16)N(0.116)S(0.003) for 500 degrees C. Bio-oils mainly contained oleic acid, 1H-indole, 2,3,5-trimethoxy toluene, toluene, (Z)-9-octadecanamide, psoralene, phenol and phenol derivatives at all pyrolysis temperatures. Both non-aromatic and aromatic hydrocarbon compounds were determined in water phase of liquid product by Headspace-GC analysis. The heating values of bio-chars were found to be similar (24MJkg(-1)) at all pyrolysis temperatures.

  19. Effect of light on contents of coumarin compounds in shoots of Ruta graveolens L. cultivated in vitro

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    Halina Ekiert


    Full Text Available Shoots of Ruta graveolens L. (Rutaceae were cultivated in stationary liquid culture under different light conditions: constant artificial light (900 lx, darkness, constant artificial light (900 ix following irradiation with UV-C light. The contents of five furanocoumarins: psoralen, bergapten, xanthotoxin, isopimpinellin and imperatorin, as well as biogenetic precursor of these metabolites, umbelliferone, were determined by HPLC method in shoots cultivated in vitro and in overground parts of plants growing in open air. It was shown that light conditions, tested in these experiments, significantly influenced contents of the metabolites in shoots cultivated in in vitro culture. Total content of the coumarin compounds in shoots cultivated under constant artificial light (900 lx was equal or higher than in plants growing under natural conditions. Therefore, it is suggested that stationary liquid shoot culture of R. graveolens. can be an alternative source for obtaining biologically active furanocoumarins.

  20. Effect of 8-MOP plus UVA treatment on survival and repair of plasmid pBR322; Efecto del tratamiento con 8-MOP mas UVA en la supervivencia y reparacion de pBR322

    Energy Technology Data Exchange (ETDEWEB)

    Bauluz, C.; Vidania, R. de


    We have studied the lethality produced in pBR322 DNA after PUVA treatment (8-MOP+UVA). As recipients, we used a collection of E. coli strains differing in their repair capacities and analysed the involvement of several DNA repair pathways in the removal of plasmid lesions. We have also studied the effect of UVA radiation alone, in order to determine more precisely the effect attributable only to psoralen molecules. Results showed a strong lethal effect derived from PUVA treatment; however, some plasmid recovery was achieved in bacterial hosts proficient in Excision repair and SOS repair. Another repair pathway, only detectable at high density of lesions, appeared to be relevant for the removal of 8-MOP:DNA adducts.(Author) 11 refs.

  1. Composição química e atividade anticolinesterásica de uma fração ativa do extrato de folhas de Citrus limon (L. Burm

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    Rusbene Bruno Fonseca de Carvalho


    Full Text Available This paper describes the chemical composition and acetylcholinesterase inhibition of an active chromatographic fraction of the EtOAc extract of Citrus limon leaves. The composition of the active fraction presented a mixture of two coumarins, 5,8-dimethoxy-psoralen and 5,7- dimethoxycoumarin, identified by ¹H and ¹³C NMR data analysis, including DEPT, COSY, HMBC and HSQC experiments. It was also demonstrated that this mixture presents qualitative and quantitative AChE inhibition. In vitro studies indicated a CE50 value of 340 µg/mL with 95% of confidence. In vivo studies (10 and 25 mg/kg revealed inhibition of 30.09 and 30.06% of AChE activity in relation to neostigmine, respectively.

  2. Simultaneous determination of sesquiterpene lactones isoalantolactone and alantolactone isomers in rat plasma by liquid chromatography with tandem mass spectrometry: application to a pharmacokinetic study. (United States)

    Guo, Chunjie; Zhang, Sixi; Teng, Shiyong; Niu, Kai


    A selective, sensitive, and accurate LC-MS/MS method for the simultaneous determination of isoalantolactone and alantolactone in rat plasma has been developed using psoralen as the internal standard. LC-MS/MS analysis was carried out on a Triple Quadrupole mass spectrometer using positive ion ESI and the selected reaction monitoring mode. The assays were linear in the range of 7.5-750 ng/mL for isoalantolactone and 5.5-550 ng/mL for alantolactone. The average recoveries in plasma samples both were better than 85%. The intra- and inter-day precision and accuracy values were found to be within the assay variability criteria limits according to the US FDA guidelines. The method was successfully applied to pharmacokinetic studies of the two structural isomers after an intravenous injection of Inula helenium formulation to rats. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. "Drug-resistant granuloma faciale": treatment with carbon dioxide-GaAs laser. (United States)

    Paradisi, Andrea; Ricci, Francesco; Sbano, Paolo


    Granuloma faciale (GF), also known as "eosinophilic granuloma," is a rare benign leukocytoclastic vasculitis which most commonly occurs on the face of middle-aged Caucasian males. Clinically, GF appears as single or multiple, slowly growing, reddish-brown papules, nodules or plaques which may be cosmetically unpleasant. Its pathogenesis is unknown and GF is notoriously resistant to treatments. Both medical (dapsone, colchicine, gold injections, isoniazid, clofazimine, corticosteroids, psoralen ultraviolet radiation, and topical tacrolimus) and surgical therapies (excision, graft, dermabrasion, argon laser, carbon dioxide laser, pulsed dye laser, cryotherapy, and electrosurgery) have been used for GF but no effective treatment has yet been found. Furthermore, the typical facial location of GF requires an acceptable cosmetic result. We report two cases of drug-resistant GF which were successfully treated with laser vaporization combining two different wavelengths: carbon dioxide (CO 2 ) 10,600 nm and GaAs 1540 nm. © 2016 Wiley Periodicals, Inc.

  4. Metronidazole-Induced Bullous Pemphigoid: A Case Report. (United States)

    Moitra, Saibal; Sen, Sukanta; Banerjee, Indranil; Sikder, Ayan; Das, Prasanta


    Bullous pemphigoid is an autoimmune cutaneous blistering disorder, the exact pathogenesis of which is still not fully elucidated. Drug-induced bullous pemphigoid eruptions are rare but have been reported earlier with the use of frusemide, psoralens, ibuprofen, galantamine hydrobromide, ACE inhibitors like captopril, spironolactone, penicillin, ampicillin, levofloxacin, penicillamine. We hereby report a case of metronidazole induced bullous pemphigoid (BP) in a 52-year-old male patient suffering from liver abscess following 4 days of drug administration. The skin biopsy findings obtained from the patient were consistent with the diagnosis of bullous pemphigoid (BP). Metronidazole was discontinued and symptomatic treatment was offered to the patient. Following withdrawal of metronidazole, the bullae subsided in the next 7-10 days without any significant residual scarring. The causality assessment performed as per the Naranjo algorithm revealed the case to be probable (Naranjo score 7).

  5. Chemical Constituents of Descurainia sophia L. and its Biological Activity

    Directory of Open Access Journals (Sweden)

    Nawal H. Mohamed


    Full Text Available Seven coumarin compounds were isolated for the first time from the aerial parts of DescurainiaSophia L. identified as scopoletine, scopoline, isoscopoline, xanthtoxol, xanthtoxin, psoralene and bergaptane.Three flavonoids namely kaempferol, quercetine and isorhamnetine and three terpenoid compounds -sitosterol-amyrine and cholesterol were also isolated and identified by physical and chemical methods; melting point, Rfvalues, UV and 1H NMR spectroscopy. Qualitative and quantitative analyses of free and protein amino acidsusing amino acid analyzer were performed. The plant contains 15 amino acids as free and protein amino acidswith different range of concentrations. Fatty acid analysis using GLC, revealed the presence of 10 fatty acids,the highest percentage was palmitic acid (27.45 % and the lowest was lauric acid (0.13%. Biological screeningof alcoholic extract showed that the plant is highly safe and has analgesic, antipyretic and anti-inflammatoryeffects.

  6. Native mitochondrial RNA-binding complexes in kinetoplastid RNA editing differ in guide RNA composition (United States)

    Madina, Bhaskara R.; Kumar, Vikas; Metz, Richard; Mooers, Blaine H.M.; Bundschuh, Ralf; Cruz-Reyes, Jorge


    Mitochondrial mRNAs in kinetoplastids require extensive U-insertion/deletion editing that progresses 3′-to-5′ in small blocks, each directed by a guide RNA (gRNA), and exhibits substrate and developmental stage-specificity by unsolved mechanisms. Here, we address compositionally related factors, collectively known as the mitochondrial RNA-binding complex 1 (MRB1) or gRNA-binding complex (GRBC), that contain gRNA, have a dynamic protein composition, and transiently associate with several mitochondrial factors including RNA editing core complexes (RECC) and ribosomes. MRB1 controls editing by still unknown mechanisms. We performed the first next-generation sequencing study of native subcomplexes of MRB1, immunoselected via either RNA helicase 2 (REH2), that binds RNA and associates with unwinding activity, or MRB3010, that affects an early editing step. The particles contain either REH2 or MRB3010 but share the core GAP1 and other proteins detected by RNA photo-crosslinking. Analyses of the first editing blocks indicate an enrichment of several initiating gRNAs in the MRB3010-purified complex. Our data also indicate fast evolution of mRNA 3′ ends and strain-specific alternative 3′ editing within 3′ UTR or C-terminal protein-coding sequence that could impact mitochondrial physiology. Moreover, we found robust specific copurification of edited and pre-edited mRNAs, suggesting that these particles may bind both mRNA and gRNA editing substrates. We propose that multiple subcomplexes of MRB1 with different RNA/protein composition serve as a scaffold for specific assembly of editing substrates and RECC, thereby forming the editing holoenzyme. The MRB3010-subcomplex may promote early editing through its preferential recruitment of initiating gRNAs. PMID:24865612

  7. GelMA-collagen blends enable drop-on-demand 3D printablility and promote angiogenesis. (United States)

    Stratesteffen, Henrike; Köpf, Marius; Kreimendahl, Franziska; Blaeser, Andreas; Jockenhoevel, Stefan; Fischer, Horst


    Effective vascularization is crucial for three-dimensional (3D) printed hydrogel-cell constructs to efficiently supply cells with oxygen and nutrients. Till date, several hydrogel blends have been developed that allow the in vitro formation of a capillary-like network within the gels but comparatively less effort has been made to improve the suitability of the materials for a 3D bioprinting process. Therefore, we hypothesize that tailored hydrogel blends of photo-crosslinkable gelatin and type I collagen exhibit favorable 3D drop-on-demand printing characteristics in terms of rheological and mechanical properties and that further capillary-like network formation can be induced by co-culturing human umbilical vein endothelial cells and human mesenchymal stem cells within the proposed blends. Gelatin was methacrylated (GelMA) at a high degree of functionalization, mixed with cells, type I collagen, and the photoinitiator Irgacure 2959 and then subsequently crosslinked with UV light. After 14 d of incubation, cells were immunofluorescently labeled (CD31) and displayed using two-photon laser scanning microscopy. Hydrogels were rheologically characterized and dispensable droplet volumes were measured using a custom built 3D drop-on-demand bioprinter. The cell viability remained high in controllable crosslinking conditions both in 2D and 3D. In general, higher UV light exposure and increased Irgacure concentration were associated with lower cell viabilities. Distinctive capillary-like structures were formed in 3D printable GelMA-collagen hydrogels. The characteristic crosslinking time for GelMA in the range of minutes was not altered when GelMA was blended with type I collagen. Moreover, the addition of collagen led to enhanced cell spreading, a shear thinning behavior of the hydrogel solution and increased the storage modulus of the crosslinked gel. We therefore conclude that GelMA-collagen hydrogels exhibit favorable biological as well as rheological properties which

  8. How insulin engages its primary binding site on the insulin receptor (United States)

    Menting, John G.; Whittaker, Jonathan; Margetts, Mai B.; Whittaker, Linda J.; Kong, Geoffrey K.-W.; Smith, Brian J.; Watson, Christopher J.; Žáková, Lenka; Kletvíková, Emília; Jiráček, Jiří; Chan, Shu Jin; Steiner, Donald F.; Dodson, Guy G.; Brzozowski, Andrzej M.; Weiss, Michael A.; Ward, Colin W.; Lawrence, Michael C.


    Insulin receptor signalling has a central role in mammalian biology, regulating cellular metabolism, growth, division, differentiation and survival1,2. Insulin resistance contributes to the pathogenesis of type 2 diabetes mellitus and the onset of Alzheimer’s disease3; aberrant signalling occurs in diverse cancers, exacerbated by crosstalk with the homologous type 1 insulin-like growth factor receptor (IGF1R)4. Despite more than three decades of investigation, the three-dimensional structure of the insulin–insulin receptor complex has proved elusive, confounded by the complexity of producing the receptor protein. Here we present the first view, to our knowledge, of the interaction of insulin with its primary binding site on the insulin receptor, on the basis of four crystal structures of insulin bound to truncated insulin receptor constructs. The direct interaction of insulin with the first leucine-rich-repeat domain (L1) of insulin receptor is seen to be sparse, the hormone instead engaging the insulin receptor carboxy-terminal α-chain (αCT) segment, which is itself remodelled on the face of L1 upon insulin binding. Contact between insulin and L1 is restricted to insulin B-chain residues. The αCT segment displaces the B-chain C-terminal β-strand away from the hormone core, revealing the mechanism of a long-proposed conformational switch in insulin upon receptor engagement. This mode of hormone–receptor recognition is novel within the broader family of receptor tyrosine kinases5. We support these findings by photo-crosslinking data that place the suggested interactions into the context of the holoreceptor and by isothermal titration calorimetry data that dissect the hormone–insulin receptor interface. Together, our findings provide an explanation for a wealth of biochemical data from the insulin receptor and IGF1R systems relevant to the design of therapeutic insulin analogues. PMID:23302862

  9. Native mitochondrial RNA-binding complexes in kinetoplastid RNA editing differ in guide RNA composition. (United States)

    Madina, Bhaskara R; Kumar, Vikas; Metz, Richard; Mooers, Blaine H M; Bundschuh, Ralf; Cruz-Reyes, Jorge


    Mitochondrial mRNAs in kinetoplastids require extensive U-insertion/deletion editing that progresses 3'-to-5' in small blocks, each directed by a guide RNA (gRNA), and exhibits substrate and developmental stage-specificity by unsolved mechanisms. Here, we address compositionally related factors, collectively known as the mitochondrial RNA-binding complex 1 (MRB1) or gRNA-binding complex (GRBC), that contain gRNA, have a dynamic protein composition, and transiently associate with several mitochondrial factors including RNA editing core complexes (RECC) and ribosomes. MRB1 controls editing by still unknown mechanisms. We performed the first next-generation sequencing study of native subcomplexes of MRB1, immunoselected via either RNA helicase 2 (REH2), that binds RNA and associates with unwinding activity, or MRB3010, that affects an early editing step. The particles contain either REH2 or MRB3010 but share the core GAP1 and other proteins detected by RNA photo-crosslinking. Analyses of the first editing blocks indicate an enrichment of several initiating gRNAs in the MRB3010-purified complex. Our data also indicate fast evolution of mRNA 3' ends and strain-specific alternative 3' editing within 3' UTR or C-terminal protein-coding sequence that could impact mitochondrial physiology. Moreover, we found robust specific copurification of edited and pre-edited mRNAs, suggesting that these particles may bind both mRNA and gRNA editing substrates. We propose that multiple subcomplexes of MRB1 with different RNA/protein composition serve as a scaffold for specific assembly of editing substrates and RECC, thereby forming the editing holoenzyme. The MRB3010-subcomplex may promote early editing through its preferential recruitment of initiating gRNAs. © 2014 Madina et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  10. Enhanced and green extraction polyphenols and furanocoumarins from Fig (Ficus carica L.) leaves using deep eutectic solvents. (United States)

    Wang, Tong; Jiao, Jiao; Gai, Qing-Yan; Wang, Peng; Guo, Na; Niu, Li-Li; Fu, Yu-Jie


    Nowadays, green extraction of bioactive compounds from medicinal plants has gained increasing attention. As green solvent, deep eutectic solvent (DES) have been highly rated to replace toxic organic solvents in extraction process. In present study, to simultaneous extraction five main bioactive compounds from fig leaves, DES was tailor-made. The tailor-made DES composed of a 3:3:3 molar ratio of glycerol, xylitol and D-(-)-Fructose showed enhanced extraction yields for five target compounds simultaneously compared with traditional methanol and non-tailor DESs. Then, the tailor-made DES based extraction methods have compared and microwave-assisted extraction was selected and optimized due to its high extraction yields with lower time consumption. The influencing parameters including extraction temperature, liquid-solid ratio, and extraction time were optimized using response surface methodology (RSM). Under optimal conditions the extraction yield of caffeoylmalic acid, psoralic acid-glucoside, rutin, psoralen and bergapten was 6.482mg/g, 16.34mg/g, 5.207mg/g, 15.22mg/g and 2.475mg/g, respectively. Macroporous resin D101 has been used to recovery target compounds with recovery yields of 79.2%, 83.4%, 85.5%, 81.2% and 75.3% for caffeoylmalic acid, psoralic acid-glucoside, rutin, psoralen and bergapten, respectively. The present study suggests that DESs are truly designer and efficient solvents and the method we developed was efficient and sustainable for extraction main compounds from Fig Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Investigation of molecular mechanisms in photodynamic action and radiobiology with nanosecond flash photolysis and pulse radiolysis. Progress report, October 1, 1980-September 30, 1981

    Energy Technology Data Exchange (ETDEWEB)

    Grossweiner, L I


    Laser flash photolysis experiments have led to a new mechanism for the ultraviolet photolysis of aqueous tryptophan (Trp), indole (Ind) and certain indole derivatives. Excitation at 265 nm leads to photoionization via a pre-fluorescent state with thermal activation. A new formula is proposed for predicting enzyme inactivation quantum yields. The predictions are in good agreement with measurements on six important enzymes at 254 nm and 280 nm. Kinetics models have been developed and tested for important stages in the photosensitization of DNA to near-ultraviolet radiation by furocoumarin compounds currently used for PUVA therapy (psoralen plus UV-A) of psoriasis and other human skin diseases. Experiments on photobinding of psoralen (Ps) and 8-methoxypsoralen (8-MOP) to calf thymus DNA are consistent with the assumption that equilibrium dark complexing of the furocoumarin to the DNA is a precondition for the formation of covalent monoadducts and cross-links. Singlet oxygen generation by furocoumarins has been investigated with liposomes and human erythrocytes (rbc). Results obtained with 3-carbethoxypsoralen (3-CPs), an experimental alternate PUVA sensitizer claimed to be non-tumorigenic, show that 3-CPs interacts with liposome and rbc membranes in the dark. Studies on photosensitization of egg lecithin liposomes by methylene blue (MB) incorporated in the membrane have led to the new result that membrane lysis is a two-stage process. The first stage induced by red light irradiation leads to membrane damage initiated by O/sub 2/*. Membrane lysis takes place in the dark, second stage under the action of mild hydrodynamic stress, such as slow gas bubbling.

  12. Mutation spectrum produced on PBR322 by 8-Methoxypsoralen plus UV-A light. Localizacion puntual de mutaciones producidas en PBR322 por tratamiento con 8-metoxipsoraleno mas luz ultravioleta-A

    Energy Technology Data Exchange (ETDEWEB)

    Bauluz, C.; Vidania, R.


    The mutagenic effect of 8-MOP+UVA (PUVA treatment) on pBR322 has been analysed by determining the frequency of mutation in the tet gene and identifying the type and position of the mutations produced inside a 276 pb-fragment (Bam-H1-SalI) of the same gene. pBR322 DNA was irradiated with UVA light in the presence of increasing concentrations of 8-Methoxypsoralen (8-MOP). The number of psoralen adducts formed in pBR322 upon that treatment ranged from 0 to 10.7 adducts per plasmid molecule. Modified DNA samples were used to transform several strains of E. Coli (differing in their repair capacities), both in constitutive conditions and after sos pre-induction by 254 nm-irradiation of cells. Mutation frequencies in the tet gene showed to increase in the wild type and uvrA strains along with the number of psoralen adduts per plasmid molecule; higher mutation frequencies were found in cells that had been previously irradiated to induce the SOS expression. Mutant plasmids were isolated from ApRTcS colonies and sequenced by the method of Maxam and Gilbert. Mutations appeared to be unique in most of the cases and were always punctual, i.e. affecting only to one base pair. The relative positions of the mutations showed a high frequency of coincidence among the sequenced fragments, indicating the existence of several DNA regions with high probability to mutated ([sup h]ot spots[sup )]. (author)

  13. Mutation spectrum produced on PBR322 by 8-Methoxypsoralen plus UV-A light; Localizacion puntual de mutaciones producidas en PBR322 por tratamiento con 8-metoxipsoraleno mas luz ultravioleta-A

    Energy Technology Data Exchange (ETDEWEB)

    Bauluz, C.; Vidania, R.


    The mutagenic effect of 8-MOP+UVA (PUVA treatment) on pBR322 has been analysed by determining the frequency of mutation in the tet gene and identifying the type and position of the mutations produced inside a 276 pb-fragment (Bam-H1-SalI) of the same gene. pBR322 DNA was irradiated with UVA light in the presence of increasing concentrations of 8-Methoxypsoralen (8-MOP). The number of psoralen adducts formed in pBR322 upon that treatment ranged from 0 to 10.7 adducts per plasmid molecule. Modified DNA samples were used to transform several strains of E. Coli (differing in their repair capacities), both in constitutive conditions and after sos pre-induction by 254 nm-irradiation of cells. Mutation frequencies in the tet gene showed to increase in the wild type and uvrA strains along with the number of psoralen adduts per plasmid molecule; higher mutation frequencies were found in cells that had been previously irradiated to induce the SOS expression. Mutant plasmids were isolated from ApRTcS colonies and sequenced by the method of Maxam and Gilbert. Mutations appeared to be unique in most of the cases and were always punctual, i.e. affecting only to one base pair. The relative positions of the mutations showed a high frequency of coincidence among the sequenced fragments, indicating the existence of several DNA regions with high probability to mutated ({sup h}ot spots{sup )}. (author)

  14. Degradable thiol-acrylate hydrogels as tunable matrices for three-dimensional hepatic culture. (United States)

    Hao, Yiting; Lin, Chien-Chi


    A degradable poly(ethylene glycol)-diacrylate (PEGDA) hydrogel system was developed using simple macromer formulations and visible light initiated thiol-acrylate photopolymerization. In addition to PEGDA, other components in this gelation system include eosin-Y as a photo-sensitizer, bi-functional thiol (dithiothreitol, DTT) as a dual-purpose co-initiator and cross-linker, and N-vinylpyrrolidone (NVP) as a co-monomer. Gelation was achieved through a mixed-mode step-chain growth polymerization mechanism under bright visible light exposure. Increasing photo-sensitizer or NVP concentrations accelerated photo-crosslinking and increased final gel stiffness. Increasing bi-functional thiol content in the prepolymer solution only increased gel stiffness to some degree. As the concentration of thiol surpassed certain range, thiol-mediated chain-transfer events caused thiol-acrylate gels to form with lower degree of cross-linking. Pendant peptide, such as integrin ligand RGDS, was more effectively immobilized in the network via a thiol-acrylate reaction (using thiol-bearing peptide Ac-CRGDS. Underline indicates cross-linkable motif) than through homo-polymerization of acrylated peptide (e.g., acryl-RGDS). The incorporation of pendant peptide comes with the expense of a lower degree of gel cross-linking, which was rectified by increasing co-monomer NVP content. Without the use of any readily degradable macromer, these visible light initiated mixed-mode cross-linked hydrogels degraded hydrolytically due to the formation of thiol-ether-ester bonds following thiol-acrylate reactions. An exponential growth relationship was identified between the hydrolytic degradation rate and bifunctional thiol content in the prepolymer solution. Finally, we evaluated the cytocompatibility of these mixed-mode cross-linked degradable hydrogels using in situ encapsulation of hepatocellular carcinoma Huh7 cells. Encapsulated Huh7 cells remained alive and proliferated as time to form cell clusters

  15. Engineered myocardium model to study the roles of HIF-1α and HIF1A-AS1 in paracrine-only signaling under pathological level oxidative stress. (United States)

    Acun, Aylin; Zorlutuna, Pinar


    Studying heart tissue is critical for understanding and developing treatments for cardiovascular diseases. In this work, we fabricated precisely controlled and biomimetic engineered model tissues to study how cell-cell and cell-matrix interactions influence myocardial cell survival upon exposure to pathological level oxidative stress. Specifically, the interactions of endothelial cells (ECs) and cardiomyocytes (CMs), and the role of hypoxia inducible factor-1α (HIF-1α), with its novel alternative regulator, HIF-1α antisense RNA1 (HIF1A-AS1), in these interactions were investigated. We encapsulated CMs in photo-crosslinkable, biomimetic hydrogels with or without ECs, then exposed to oxidative stress followed by normoxia. With precisely controlled microenvironment provided by the model tissues, cell-cell interactions were restricted to be solely through the secreted factors. CM survival after oxidative stress was significantly improved, in the presence of ECs, when cells were in the model tissues that were functionalized with cell attachment motifs. Importantly, the cardioprotective effect of ECs was reduced when HIF-1α expression was knocked down suggesting that HIF-1α is involved in cardioprotection from oxidative damage, provided through secreted factors conferred by the ECs. Using model tissues, we showed that cell survival increased with increased cell-cell communication and enhanced cell-matrix interactions. In addition, whole genome transcriptome analysis showed, for the first time to our knowledge, a possible role for HIF1A-AS1 in oxidative regulation of HIF-1α. We showed that although HIF1A-AS1 knockdown helps CM survival, its effect is overridden by CM-EC bidirectional interactions as we showed that the conditioned media taken from the CM-EC co-cultures improved CM survival, regardless of HIF1A-AS1 expression. Cardiovascular diseases, most of which are associated with oxidative stress, is the most common cause of death worldwide. Thus, understanding

  16. Mitigating Scarring and Inflammation during Corneal Wound Healing using Nanofiber-Hydrogel Scaffolds (United States)

    Fu, Amy

    Due to the universal lack of donor tissue, there has been emerging interest in engineering materials to stimulate living cells to restore the features and functions of injured organs. We are particularly interested in developing materials for corneal use, where the necessity to maintain the tissue's transparency presents an additional challenge. Every year, there are 1.5 -- 2 million new cases of monocular blindness due to irregular healing of corneal injuries, dwarfing the approximately 150,000 corneal transplants performed. The large gap between the need and availability of cornea transplantation motivates us to develop a wound-healing scaffold that can prevent corneal blindness. To develop such a scaffold, it is necessary to regulate the cells responsible for repairing the damaged cornea, namely myofibroblasts, which are responsible for the disordered and non-refractive index matched scar that leads to corneal blindness. Using in vitro assays, we identified that protein nanofibers of certain orientation can promote cell migration and modulate the myofibroblast phenotype. The nanofibers are also transparent, easy to handle and non-cytotoxic. To adhere the nanofibers to a wound bed, we examined the use of two different in situ forming hydrogels: an artificial extracellular matrix protein (aECM)-based gel and a photo-crosslinkable heparin-based gel. Both hydrogels can be formed within minutes, are transparent upon gelation and are easily tunable. Using an in vivo mouse model for epithelial defects, we show that our corneal scaffolds (nanofibers together with hydrogel) are well-tolerated (no inflammatory response or turbidity) and support epithelium regrowth. We developed an ex vivo corneal tissue culture model where corneas that are wounded and treated with our scaffold can be cultured while retaining their ability to repair wounds for up to 21 days. Using this technique, we found that the aECM-based treatment induced a more favorable wound response than the

  17. Fc-specific biotinylation of antibody using an engineered photoactivatable Z–Biotin and its biosensing application

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Hong-Ming; Bao, Ru-Meng; Yu, Chang-Mei; Lv, Yan-Na; Zhang, Wei-Fen; Tang, Jin-Bao, E-mail:


    The development of a site-specific and covalent attachment methodology is crucial for antibody–biotin conjugates to preserve the antigen-binding ability of antibodies and yield homogeneous products. In this study, an engineered photoactivatable Z-domain variant [an UV-active amino acid benzoylphenylalanine (Bpa) was genetically incorporated into the Z-domain] carrying one biotin molecule (Z{sub Bpa}–Biotin) was prepared by employing aminoacyl-tRNA synthetase/suppressor tRNA and Avitag/BirA techniques. The site-specific and covalent attachment of IgG–biotin conjugates, viz. photo-biotinylated IgG, was successfully achieved after UV exposure by combining the inherent Fc-binding capability of the Z-domain with the formation of covalent bond by the photo-crosslinker. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis assay showed that more than 90% of IgGs conjugated with Z{sub Bpa}–Biotin molecules suffered 3 h UV irradiation. Further pepsin digestion analysis confirmed that the Z{sub Bpa}–Biotin was conjugated to the Fc fragment of IgG without interference. We took the tumor biomarker carcinoembryoic antigen (CEA) as model to evaluate the detection efficiency of the site-specific photo-biotinylated IgG in biosensing application using surface plasmon resonance (SPR) technology. The photo-biotinylated IgG coated surface gave a limit of detection (LOD) of 2 ng mL{sup -1}, is 5-fold lower than that of the randomly NHS-biotinylated IgG (10 ng mL{sup -1}). Given that the (strept)avidin–biotin complex is extensively used in immunoassays, the proposed method for biotinylated IgG provides a powerful approach to further expand related applications. - Highlights: • A photoactivable Z{sub Bpa}–Biotin was fabricated by aaRS/tRNA and Avitag/BirA techniques. • A approach for Fc-specific photo-biotinylated IgG via Z{sub Bpa}–Biotin was proposed. • The photo-biotinylated IgG was used to fabricate an immunosensor for detecting CEA. • It gave a LOD

  18. Nuclear envelope breakdown is under nuclear not cytoplasmic control in sea urchin zygotes. (United States)

    Sluder, G; Thompson, E A; Rieder, C L; Miller, F J


    Nuclear envelope breakdown (NEB) and entry into mitosis are though to be driven by the activation of the p34cdc2-cyclin B kinase complex or mitosis promoting factor (MPF). Checkpoint control mechanisms that monitor essential preparatory events for mitosis, such as DNA replication, are thought to prevent entry into mitosis by downregulating MPF activation until these events are completed. Thus, we were surprised to find that when pronuclear fusion in sea urchin zygotes is blocked with Colcemid, the female pronucleus consistently breaks down before the male pronucleus. This is not due to regional differences in the time of MPF activation, because pronuclei touching each other break down asynchronously to the same extent. To test whether NEB is controlled at the nuclear or cytoplasmic level, we activated the checkpoint for the completion of DNA synthesis separately in female and male pronuclei by treating either eggs or sperm before fertilization with psoralen to covalently cross-link base-paired strands of DNA. When only the maternal DNA is cross-linked, the male pronucleus breaks down first. When the sperm DNA is cross-linked, male pronuclear breakdown is substantially delayed relative to female pronuclear breakdown and sometimes does not occur. Inactivation of the Colcemid after female NEB in such zygotes with touching pronuclei yields a functional spindle composed of maternal chromosomes and paternal centrosomes. The intact male pronucleus remains located at one aster throughout mitosis. In other experiments, when psoralen-treated sperm nuclei, over 90% of the zygote nuclei do not break down for at least 2 h after the controls even though H1 histone kinase activity gradually rises close to, or higher than, control mitotic levels. The same is true for normal zygotes treated with aphidicolin to block DNA synthesis. From these results, we conclude that NEB in sea urchin zygotes is controlled at the nuclear, not cytoplasmic, level, and that mitotic levels of

  19. Successful treatment of palmoplantar pustulosis with isotretinoin. (United States)

    Wilken, Reason; Sharma, Ajay; Patel, Forum; Maverakis, Emanual


    Variably considered as a localized subtype of pustular psoriasis, palmoplantar pustulosis (PPP) is commonly treated with topical steroids, acitretin, and local phototherapy with oral or topical psoralen (PUVA). The utility of acitretin for PPP is limited by adverse effects such as myalgias and an extended risk of teratogenicity in female patients. Isotretinoin is a more tolerable retinoid with a shorter teratogenic window, but to date its effectiveness in PPP has not been reported. Herein we present two patients with PPP who responded well to isotretinoin treatment. Two patients with PPP refractory to topical therapies were started on acitretin. Both patients developed adverse effects (including headache, myalgias, and mood alterations) leading to acitretin discontinuation. Isotretinoin monotherapy was started in one patient resulting in significant clearing of palmar plaques and scale, and the addition of isotretinoin to UVA therapy resulted in near-complete clearing of recalcitrant plantar plaques in the second patient. Acitretin represents an important treatment for PPP, but is limited by adverse effects and extended teratogenicity. Our experience supports the utility of isotretinoin as a potential therapeutic alternative, which may be particularly beneficial in patients who are poor candidates for or unable to tolerate acitretin therapy.

  20. Development of an LC/MS/MS method in order to determine arctigenin in rat plasma: its application to a pharmacokinetic study. (United States)

    Zou, Quanfei; Gu, Yuan; Lu, Rong; Zhang, Tiejun; Zhao, Guang-Rong; Liu, Changxiao; Si, Duanyun


    In this study, a simple and sensitive LC/MS/MS method was developed and validated for the determination of arctigenin in rat plasma. The MS detection was performed using multiple reaction monitoring at the transitions of m/z 373.2 → 137.3 for arctigenin and m/z 187.1 → 131.0 for psoralen (internal standard) with a Turbo IonSpray electrospray in positive mode. The calibration curves fitted a good linear relationship over the concentration range of 0.2-500 ng/mL. It was found that arctigenin is not stable enough at both room temperature and -80 °C unless mixed with methanol before storage. The validated LC/MS/MS method was successfully applied for the pharmacokinetic study of arctigenin in rats. After intravenous injection of 0.3 mg/kg arctigenin injection to rats, the maximum concentration, half-life and area under the concentration-time curve were 323 ± 65.2 ng/mL, 0.830 ± 0.166 and 81.0 ± 22.1 h ng/mL, respectively. Copyright © 2013 John Wiley & Sons, Ltd.

  1. Incidence and prevalence of psoriasis in Denmark

    DEFF Research Database (Denmark)

    Egeberg, Alexander; Skov, Lone; Gislason, Gunnar H.


    The incidence and temporal trends of psoriasis in Denmark between 2003 and 2012 were examined. There was a female predominance ranging between 50.0% (2007) and 55.4% (2009), and the mean age at time of diagnosis was 47.7-58.7 years. A total of 126,055 patients with psoriasis (prevalence 2.2%) were...... identified. Incidence rates of psoriasis (per 100,000 person years) ranged from 107.5 in 2005 to a peak incidence of 199.5 in 2010. Incidence rates were higher for women, and patients aged 60-69 years, respectively. Use of systemic non-biologic agents, i.e. methotrexate, cyclosporine, retinoids, or psoralen...... plus ultraviolet A (PUVA) increased over the study course, and were used in 15.0% of all patients. Biologic agents (efalizumab, etanercept, infliximab, adalimumab, or ustekinumab) were utilized in 2.7% of patients. On a national level, incidence of psoriasis fluctuated during the 10- year study course...

  2. A phase II placebo-controlled study of photodynamic therapy with topical hypericin and visible light irradiation in the treatment of cutaneous T-cell lymphoma and psoriasis. (United States)

    Rook, Alain H; Wood, Gary S; Duvic, Madeleine; Vonderheid, Eric C; Tobia, Alfonso; Cabana, Bernard


    Hypericin is a known photodynamic agent that has been demonstrated to induce apoptosis in normal and malignant B and T lymphocytes, and has potential to treat benign and malignant disorders of the skin, including psoriasis and cutaneous T-cell lymphoma. We wished to test whether topical hypericin was an effective, safe, and well-tolerated therapy for patch or plaque phase mycosis fungoides and for plaque psoriasis. We conducted a phase II placebo-controlled clinical study in patients who had either patch or plaque phase mycosis fungoides or plaque type psoriasis vulgaris. Representative lesions were treated twice weekly for 6 weeks with topically applied hypericin or placebo followed 24 hours later by exposure to visible light at 8 to 20 J/cm(2). After 6 weeks of twice-weekly therapy, several concentrations of hypericin resulted in the significant improvement of treated skin lesions among the majority of patients with cutaneous T-cell lymphoma and psoriasis whereas the placebo vehicle was ineffective. The clinical trial involved a small number of patients. Overall, the data from this study support the conclusion that topical hypericin/visible light photodynamic therapy is an effective and well-tolerated alternative to standard psoralen plus ultraviolet A treatment of these disorders. Copyright © 2010 American Academy of Dermatology, Inc. Published by Mosby, Inc. All rights reserved.

  3. Hypomutability in Fanconi anemia cells is associated with increased deletion frequency at the HPRT locus

    Energy Technology Data Exchange (ETDEWEB)

    Papadopoulo, D.; Guillouf, C.; Moustacchi, E. (Unite Recherche Associee due Centre National del La Recherche Scientifique, Paris (France)); Mohrenweiser, H. (Lawrence Livermore National Lab., CA (United States))


    Fanconi anemia (FA) is an inherited human disorder associated with a predisposition to cancer and characterized by anomalies in the processing of DNA cross-links and certain monoadducts. The authors reported previously that the frequency of psoralen-photoinduced mutations at the HPRT locus is lower in FA cells than in normal cells. This hypomutability is shown here to be associated with an increased frequency of deletions in the HPRT gene when either a mixture of cross-links and monoadducts or monoadducts alone are induced. Molecular analysis of mutants in the HPRT gene was carried out. In normal cells the majority of spontaneous and induced mutants are point mutations whereas in FA deletion mutations predominate. In that case a majority of mutants were found to lack individual exons or small clusters of exons whereas in normal cells large (complete or major gene loss) and small deletions are almost equally represented. Thus they propose that the FA defect lies in a mutagenic pathway that, in normal cells, involves by passing lesions and subsequent gap filling by a recombinational process during replication.

  4. Granulomatous slack skin - seven years follow-up of a case with features of "parapsoriasis en plaques", muscle involvement, granulomatous vasculitis, and necrobiotic changes. (United States)

    ChiriŢă, Aurel Doru; Mărgăritescu, Irina


    Granulomatous slack skin (GSS) represents an extremely rare variant of mycosis fungoides with only 70 cases reported in the literature to date. It is characterized clinically by the occurrence of bulky, pendulous skinfolds, usually located in flexural areas and histologically by an infiltrate composed of small neoplastic T-lymphocytes joined by granulomatous inflammation with scattered multinucleated giant cells containing nuclei arranged in a wreath-like fashion. Since its first description, very rare cases of GSS with muscle involvement, large vessels involvement, or necrobiotic changes have been reported. We present an extraordinary case of GSS with all these unusual features developing in the lesions of the same patient. The long follow-up of seven years allowed us to document the evolution of each lesion. Some lesions appeared and evolved in a manner very reminiscent of those of "parapsoriasis en plaques", others were classical GSS lesions, and still others developed large ulcerated lesions. These ulcerated lesions consistently failed to respond to conventional wound therapy, skin directed therapy [retinoids + psoralen combined with ultraviolet A (PUVA)-therapy], and interferon-alpha therapy. Remarkably, the ulcers completely healed when systemic corticosteroids were added. We hence postulate that the ulcers appeared because of large vessel vasculitis rather than tumoral direct destruction.

  5. Effects of PUVA on the eye

    Energy Technology Data Exchange (ETDEWEB)

    Backman, H.A.


    Psoriasis is a common skin disease which may be treated with 8-methoxy psoralen and long-wave ultraviolet light (PUVA). Eye protection is provided during and after treatment to prevent the development of photokeratitis and cataracts. Fifteen patients, treated with medication and ultraviolet A (UVA) had an initial complete eye examination and a repeat examination after each treatment. No patients developed cataracts but almost one-half of the patients had a mild form of photokeratoconjunctivitis. The ocular manifestations included photophobia, conjunctivitis, keratitis, and dry eyes. Tear break-up time was reduced significantly immediately after treatment for two patients but returned to normal 8 hr later. Dermatologists who employ PUVA treatments should be concerned about photokeratoconjunctivitis and the dry-eye ocular manifestations included photophobia, conjunctivitis, keratitis, and dry eyes. Tear break-up time was reduced significantly immediately after treatment for two patients but returned to normal 8 hr later. Dermatologists who employ PUVA treatments should be concerned about photokeratoconjunctivitis and the dry-eye ocular manifestations included photophobia, conjunctivitis, keratitis, and dry eyes. Tear break-up time was reduced significantly immediately after treatment for two patients but returned to normal 8 hr later. Dermatologists who employ PUVA treatments should be concerned about photokeratoconjunctivitis and the dry-eye syndrome.

  6. Chinese single herbs and active ingredients for postmenopausal osteoporosis: From preclinical evidence to action mechanism. (United States)

    Lin, Jing; Zhu, Jun; Wang, Yan; Zhang, Na; Gober, Hans-Jürgen; Qiu, Xuemin; Li, Dajin; Wang, Ling


    Postmenopausal osteoporosis is a systemic metabolic skeletal disease generally ascribable to a dearth of estrogen. Whether traditional Chinese medicine is effective in management of postmenopausal osteoporosis remains unclear. This article reviews the experimental evidence of both in vitro and in vivo preclinical studies with the theme of the application of Chinese single herbs and active ingredients in postmenopausal osteoporosis. It includes three single herbs (Herba Epimedium, Rhizoma Drynariae, and Salvia miltiorrhiza) and eight active ingredients (saikosaponins, linarin, echinacoside, sweroside, psoralen, poncirin, vanillic acid, and osthole). The experimental studies indicated their potential use as treatment for postmenopausal osteoporosis and investigated the underlying mechanisms including osteoprotegerin/receptor activator of nuclear factor κB ligand (OPG/RANKL), extracellular-signal-regulated kinase/c-Jun N terminal kinase/mitogen-activated protein kinase (ERK/JNK/MAPK), estrogen receptor (ER), bone morphogenetic protein (BMP), transforming growth factor (TGF)-β, Wnt/β-catenin, and Notch signaling pathways. This review contributes to a better understanding of traditional Chinese medicine and provides useful information for the development of more effective anti-osteoporosis drugs.

  7. Thorough investigation of the oxygen heterocyclic fraction of lime (Citrus aurantifolia (Christm.) Swingle) juice. (United States)

    Costa, Rosaria; Russo, Marina; De Grazia, Selenia; Grasso, Elisa; Dugo, Paola; Mondello, Luigi


    Reversed-phase-HPLC analysis by means of superficially porous silica particle columns (fused-core) was applied to the investigation of flavonoids, coumarins, and psoralens in lime juice samples. Hesperidin (367.0 ± 16.0 ppm) and eriocitrin (148.0 ± 7.9 ppm) were the most abundant flavonoids. Fifteen coumarins and furocoumarins were determined, including bergamottin (29.6 ± 1.1 ppm), 5-geranyloxy-7-methoxycoumarin (16.5 ± 0.6 ppm), and oxypeucedanin hydrate (9.9 ± 0.5 ppm) as predominant compounds. These molecules are today well known for their beneficial effects on human health. As a consequence, the present study, beyond investigating for the first time the chemical composition of lime juice, highlights also its health-promoting qualities, due to its content of flavonoids and coumarins. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Zeolite scaffolds for cultures of human breast cancer cells. Part II: Effect of pure and hybrid zeolite membranes on neoplastic and metastatic activity control. (United States)

    Tavolaro, Palmira; Martino, Guglielmo; Andò, Sebastiano; Tavolaro, Adalgisa


    This work is focused on the response of two invasive phenotypes of human breast cancer cells, MCF-7 and MDA-MB-231, grown on synthesized zeolite scaffolds in order to study the influence of those biomaterials in controlled conditions with and without anti-tumoral drug treatments. Our research was directed to the use of doxorubicin (DOX) and bergapten (5-MOP). The former is broadly considered the most active single agent available for the treatment of breast cancer, the second is a natural psoralen with an apoptotic effect. The results indicate that both drugs inhibit the cell viability of all cell lines grown on all zeolite scaffolds and that all Pure Zeolite Membranes are more responsive with respect to all Mixed Matrix Membranes. Moreover, the results after treatment with DOX at a concentration of 7.4μM for 24h, show that the expression of the matrix metalloproteinases (MMP-2 and MMP-9) is greatly reduced in both cell lines, especially in those adherent on Pure Zeolite Scaffolds. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Fetal Hemoglobin Inducers from the Natural World: A Novel Approach for Identification of Drugs for the Treatment of {beta}-Thalassemia and Sickle-Cell Anemia. (United States)

    Bianchi, Nicoletta; Zuccato, Cristina; Lampronti, Ilaria; Borgatti, Monica; Gambari, Roberto


    The objective of this review is to present examples of lead compounds identified from biological material (fungi, plant extracts and agro-industry material) and of possible interest in the field of a pharmacological approach to the therapy of beta-thalassemia using molecules able to stimulate production of fetal hemoglobin (HbF) in adults. Concerning the employment of HbF inducers as potential drugs for pharmacological treatment of beta-thalassemia, the following conclusions can be reached: (i) this therapeutic approach is reasonable, on the basis of the clinical parameters exhibited by hereditary persistence of fetal hemoglobin patients, (ii) clinical trials (even if still limited) employing HbF inducers were effective in ameliorating the symptoms of beta-thalassemia patients, (iii) good correlation of in vivo and in vitro results of HbF synthesis and gamma-globin mRNA accumulation indicates that in vitro testing might be predictive of in vivo responses and (iv) combined use of different inducers might be useful to maximize HbF, both in vitro and in vivo. In this review, we present three examples of HbF inducers from the natural world: (i) angelicin and linear psoralens, contained in plant extracts from Angelica arcangelica and Aegle marmelos, (ii) resveratrol, a polyphenol found in grapes and several plant extracts and (iii) rapamycin, isolated from Streptomyces hygroscopicus.

  10. Inactivated simian immunodeficiency virus vaccine failed to protect rhesus macaques from intravenous or genital mucosal infection but delayed disease in intravenously exposed animals

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    Sutjipto, S.; Pedersen, N.C.; Miller, C.J.; Gardner, M.B.; Hanson, C.V.; Gettie, A.; Jennings, M.; Higgins, J.; Marx, P.A. (Univ. of California, Davis (USA))


    Eight rhesus macaques were immunized four times over a period of 8 months with a psoralen-UV-light-inactivated whole simian immunodeficiency virus vaccine adjuvanted with threonyl muramyl dipeptide. Eight unvaccinated control animals received adjuvant alone. Only the vaccinated animals made antibodies before challenge exposure to the viral core and envelope as determined by Western blotting (immunoblotting) and virus-neutralizing antibodies. Ten days after the final immunization, one-half of the vaccinated and nonvaccinated monkeys were challenged exposed intravenously (i.v.) and one-half were challenge exposed via the genital mucosa with virulent simian immunodeficiency virus. All of the nonvaccinated control monkeys became persistently infected. In spite of preexisting neutralizing antibodies and an anamnestic antibody response, all of the immunized monkeys also became persistently infected. However, there was evidence that the clinical course in immunized i.v. infected animals was delayed. All four mock-vaccinated i.v. challenge-exposed animals died with disease from 3 to 9 months postchallenge. In contrast, only one of four vaccinated i.v. challenge-exposed monkeys had died by 11 months postchallenge.

  11. Chromatin structure of ribosomal RNA genes in dipterans and its relationship to the location of nucleolar organizers.

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    Christiane Rodriguez Gutierrez Madalena

    Full Text Available Nucleoli, nuclear organelles in which ribosomal RNA is synthesized and processed, emerge from nucleolar organizers (NORs located in distinct chromosomal regions. In polytene nuclei of dipterans, nucleoli of some species can be observed under light microscopy exhibiting distinctive morphology: Drosophila and chironomid species display well-formed nucleoli in contrast to the fragmented and dispersed nucleoli seen in sciarid flies. The available data show no apparent relationship between nucleolar morphology and location of NORs in Diptera. The regulation of rRNA transcription involves controlling both the transcription rate per gene as well as the proportion of rRNA genes adopting a proper chromatin structure for transcription, since active and inactive rRNA gene copies coexist in NORs. Transcription units organized in nucleosomes and those lacking canonical nucleosomes can be analyzed by the method termed psoralen gel retarding assay (PGRA, allowing inferences on the ratio of active to inactive rRNA gene copies. In this work, possible connections between chromosomal location of NORs and proportion of active rRNA genes were studied in Drosophila melanogaster, and in chironomid and sciarid species. The data suggested a link between location of NORs and proportion of active rRNA genes since the copy number showing nucleosomal organization predominates when NORs are located in the pericentric heterochromatin. The results presented in this work are in agreement with previous data on the chromatin structure of rRNA genes from distantly related eukaryotes, as assessed by the PGRA.

  12. Photocarcinogenicity of selected topically applied dermatological drugs: calcineurin inhibitors, corticosteroids, and vitamin D analogs

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    Catharina Margrethe Lerche


    Full Text Available Topical therapies constitute the mainstay of dermatological treatments for skin disorders, such as atopic dermatitis, contact dermatitis, psoriasis, or acne. Since some of these diseases are often chronic, treatment duration may last for years and may even last the patient’s entire lifetime. Obviously, such long-term therapy may raise safety concerns, which also include the potential photocarcinogenic effect. Most patients are exposed to ultraviolet radiation (UVR during leisure, work, vacations, or in tanning beds. Additionally, the patients may receive UVR via UVB phototherapy or psoralens plus UVA radiation (PUVA. The use of immunosuppressant’s, such as corticosteroids and calcineurin inhibitors, has markedly increased. Patients with skin diseases have benefited from both systemic and topical treatment of both new and established drugs. The issue of a black box warning by the US Food and Drug Administration has increased concerns about photocarcinogenesis, which raises the question: “Are these drugs safe?” This review focuses on the mechanism of action and photocarcinogenic potential of commonly used topical treatments, such as corticosteroids, calcineurin inhibitors, and vitamin D analogs.

  13. Preliminary In Vitro and In Vivo Evaluation of Antidiabetic Activity of Ducrosia anethifolia Boiss. and Its Linear Furanocoumarins

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    Nagwa M. M. Shalaby


    Full Text Available Aim. Ducrosia anethifolia is used as flavoring additive. There have been little detailed phytochemical reports on this genus and the antidiabetic activity of this plant is not yet evaluated. Method. Structure of compounds was deduced by spectroscopic analyses. Preliminary in vitro evaluation of the antidiabetic activity of crude extract and its furanocoumarins was carried out (α-amylase, α-glucosidase, and β-galactosidase. The in vivo activity was investigated by measuring some oxidative stress markers. Biomarkers of liver injury and kidney were also determined. Results. Eight linear furanocoumarins, psoralen, 5-methoxypsoralen, 8-methoxypsoralen, imperatorin, isooxypeucedanin, pabulenol, oxypeucedanin methanolate, oxypeucedanin hydrate, and 3-O-glucopyranosyl-β-sitosterol, were isolated. All compounds were reported for the first time from the genus Ducrosia except pabulenol. The blood glucose level, liver function enzymes, total protein, lipid, and cholesterol levels were significantly normalized by extract treatment. The antioxidant markers, glucolytic, and gluconeogenic enzymes were significantly ameliorated and the elevated level of kidney biomarkers in the diabetic groups was restored. The compounds showed inhibitory activity in a concentration dependant manner. Imperatorin and 5-methoxypsoralen showed the most potent inhibiting power. Conclusion. D. anethifolia extract showed hypoglycemic, hypolipidemic, and antioxidant effect as well as ameliorating kidney function. This extract and some linear furanocoumarins exhibited carbohydrate metabolizing enzymes inhibitory effect.

  14. Phototoxicity, Pseudoporphyria, and Photo-onycholysis Due to Voriconazole in a Pediatric Patient With Leukemia and Invasive Aspergillosis. (United States)

    Willis, Zachary I; Boyd, Alan S; Di Pentima, M Cecilia


    Voriconazole is a triazole antifungal agent superior to amphotericin B in the treatment of invasive aspergillosis. It is generally well tolerated and has excellent oral bioavailability, providing significant benefit in the treatment of invasive fungal infections. There have been numerous reports of dermatologic reactions to this agent, including erythroderma, cheilitis, Stevens-Johnson syndrome, discoid lupus erythematosus, pseudoporphyria, squamous cell carcinoma, and photosensitivity reactions. Pseudoporphyria, a dermatologic condition mimicking porphyria cutanea tarda, has been described as an adverse effect of voriconazole use. Clinical findings include photosensitivity, vesicles, bullae, milia, and scarring in sun-exposed areas. Photo-onycholysis is a phenomenon of nail discoloration and onycholysis that has been described in the setting of a phototoxic drug reaction and pseudoporphyria. Implicated drugs have most commonly been tetracyclines, fluoroquinolones, and psoralens; others have been reported as well. We report a case of a pediatric patient with leukemia who developed symptoms consistent with pseudoporphyria and later photo-onycholysis while being treated with voriconazole. To our knowledge, this is the first reported case of pseudoporphyria due to voriconazole in a pediatric patient and the first reported case of photo-onycholysis as a consequence of voriconazole use. © The Author 2014. Published by Oxford University Press on behalf of the Pediatric Infectious Diseases Society. All rights reserved. For Permissions, please e-mail:

  15. Genital squamous cell carcinoma in men treated by photochemotherapy. A cancer registry-based study from 1978 to 1998. (United States)

    Aubin, F; Puzenat, E; Arveux, P; Louvat, P; Quencez, E; Humbert, P


    One single report from the U.S. 16-centre-trial indicated that psoralen and ultraviolet A radiation (PUVA) therapy may induce an increased risk of genital tumours in men, and protection of the genital area is, therefore, recommended. To evaluate the relevance of this risk in routine clinical practice. Two groups of patients were included in a 1978-98 retrospective study. Case records of men with genital squamous cell carcinoma (SCC) identified from the Cancer Registry of the Doubs area of France were examined for a history of PUVA therapy, topical tar treatment, psoriasis, human papillomavirus infection or genital dermatitis. In addition, all the dermatologists of the Doubs area (in public and private practice) using PUVA therapy were asked to provide information on the number of patients having received PUVA therapy and whether the genital area was exposed during treatment. Between 1978 and 1998, among the 48 men who had developed a genital SCC in the Doubs area, only one had a history of intensive PUVA therapy. About 150,000 treatments with PUVA therapy had been performed by 15 dermatologists in the Doubs area for 5400 patients since 1978. No case of genital SCC had been reported, despite the fact that the genital area had not been protected during UVA exposure. Although retrospective, our study demonstrates that the occurrence of genital SCC in men treated with PUVA therapy is a very rare event in common dermatological practice.

  16. Antidiabetics and diuretics show phototoxicity in HaCaT cells (United States)

    Selvaag, Edgar; Petersen, Anita B.; Gniadecki, Robert; Thorn, Tine; Wulf, Hans Christian


    The antidiabetics tolbutamide, glibenclamide, and glipizide, and the diuretics bendroflumethiazide, butizide, furosemide, hydrochlorothiazide, and trichlormethiazide were investigated for potential phototoxicity in the HaCaT cell line. The cells were incubated with the drugs and then exposed to UVA1 irradiation. The effects of the antioxidants L-ascorbic acid, and (alpha) -tocopherol on oxidative DNA damage were assessed. Bendroflumethiazide, furosemide, hydrochlorothiazide, trichlormethiazide, or tolbutamide induced dose-dependent phototoxicity. Cells incubated with bendroflumethiazide, tolbutamide, and glibenclamide, and irradiated with UVA1 demonstrated an increased oxidative DNA damage. Pre-treatment with L-ascorbic acid, or (alpha) -tocopherol, suppressed the UVA-induced DNA damage in cells incubated with 1 mM of bendroflumethiazide, furosemide, glibenclamide, glipizide, tolbutamide, and trichloromethiazide, further implying the involvement of reactive oxygen species in the phototoxic DNA damage. These results may indicate a link between phototoxic and photocancerogenic potential of the sulfonamide-derived oral antidiabetic and diuretic drugs, as it has previously been recognized for psoralen, chlorpromazine, and fluoroquinolones. Excessive exposure to UV light may be deleterious for patients treated with these drugs.

  17. Current knowledge in Polypodium leucotomos effect on skin protection. (United States)

    Palomino, Olga María


    This article provides an overview of pharmacology, toxicity, pharmacokinetics and clinical data of Polypodium leucotomos L. (PL). PL aerial part has proven to exert antioxidant, photoprotective and immunomodulatory activities; its mechanism of action is complex and includes several activities: (1) PL diminishes the production of reactive oxygen and nitrogen species (ROS, RNS); (2) PL inhibits the photoisomerization of trans-urocanic acid (t-UCA); (3) PL inhibits apoptosis induced by ultraviolet radiation; (4) PL prevents damage to genetic material and (5) PL enhances DNA repair. PL is not mutagenic and does not induce acute or chronic toxicity. Its biological effects have been proved in cell cultures, animal models, murine models and in human beings. Photoprotective activity has been assessed in healthy volunteers as well as in patients suffering from several cutaneous diseases such as vitiligo, psoriasis, idiopathic photodermatosis or melasma. PL results to be an efficient treatment especially for sensitive cutaneous phototypes and adds extra protection when ultraviolet radiation (UVR) exposure cannot be avoided, such as wide or narrow band UVB phototherapy or treatment with psoralens plus UVA exposure radiation.

  18. Drug-induced nail disorders. (United States)


    Nail disorders are defined according to their appearance and the part of the nail affected: the nail plate, the tissues that support or hold the nail plate in place, or the lunula. The consequences of most nail disorders are purely cosmetic. Other disorders, such as ingrown nails, inflammation, erythema, abscesses or tumours, cause functional impairment or pain. The appearance of the lesions is rarely indicative of their cause. Possible causes include physiological changes, local disorders or trauma, systemic conditions, toxic substances and drugs. Most drug-induced nail disorders resolve after discontinuation of the drug, although complete resolution sometimes takes several years. Drugs appear to induce nail disorders through a variety of mechanisms. Some drugs affect the nail matrix epithelium, the nail bed or the nail folds. Some alter nail colour. Other drugs induce photosensitivity. Yet others affect the blood supply to the nail unit. Nail abnormalities are common during treatment with certain cytotoxic drugs: taxanes, anthracyclines, fluorouracil, EGFR, tyrosine kinase inhibitors, etc. Some drugs are associated with a risk of serious and painful lesions, such as abscesses. When these disorders affect quality of life, the benefits of withdrawing the drug must be weighed against the severity of the condition being treated and the drug's efficacy, taking into account the harm-benefit balance of other options. Various anti-infective drugs, including tetracyclines, quinolones, clofazimine and zidovudine, cause the nail plate to detach from the nail bed after exposure to light, or cause nail discoloration. Psoralens and retinoids can also have the same effects.

  19. An UPLC-MS/MS method for simultaneous quantitation of two coumarins and two flavonoids in rat plasma and its application to a pharmacokinetic study of Wikstroemia indica extract. (United States)

    Wei, Lan; Wang, Xiaobo; Zhang, Peng; Sun, Yangyang; Jia, Lili; Zhao, Jingxin; Dong, Shikai; Sun, Lixin


    In this study, an ultra performance liquid chromatography tandem mass spectrometry method (UPLC-MS/MS) was developed for simultaneous determination of umbelliferone, apigenin, daphnoretin and genkwanin in total (free and conjugated) forms in rat plasma using psoralen as internal standard. Plasma samples were protein precipitated with acetonitrile followed by liquid-liquid extracted with ethyl acetate. Four ingredients were separated on an Acquity UPLC(®) BEH C18 column using gradient elution with the mobile phase consisting of 0.1% formic acid aqueous solution and acetonitrile, and detected by positive ion electrospray ionization (ESI) in multiple reaction monitoring mode (MRM). The method was linear for all analytes over investigated ranges with all correlation coefficients greater than 0.99. The validated lower limit of quantification was 3ng/mL for umbelliferone, 3ng/mL for apigenin, 12ng/mL for daphnoretin and 2ng/mL for genkwanin, respectively. Intra- and inter-day precisions (RSD%) were less than 15% and accuracy (RE%) ranged from -1.1% to 15%. The mean absolute extraction recoveries of analytes and IS from rat plasma were all more than 75%. The validated method was firstly and successfully applied to investigate the pharmacokinetics of four chemical ingredients after oral administration of Wikstroemia indica extract (WIE) to rats. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Simultaneous assessment of absorption characteristics of coumarins from Angelicae Pubescentis Radix: In vitro transport across Caco-2 cell and in vivo pharmacokinetics in rats after oral administration. (United States)

    Yang, Yan-Fang; Zhang, Lei; Zhang, You-Bo; Yang, Xiu-Wei


    Angelicae Pubescentis Radix (APR), a well-known traditional Chinese medicine, is widely used for the treatments of rheumatism and headache for centuries. To assess the absorption characteristics of coumarins from APR, a sensitive and reliable UPLC-MS/MS method was established for the simultaneous determination of sixteen coumarins from APR, including psoralen, xanthotoxin, bergapten, bergaptol, isoimperatorin, imperatorin, columbianetin, columbianetin acetate, columbianadin, oxypeucedanin hydrate, angelol B, umbelliferone, scopoletin, osthole, meranzin hydrate and nodakenetin. The specificity, linearity, sensitivity, precision, accuracy, recovery, matrix effect and stability of the method were all validated to be satisfactory. The method was then applied to the in vitro transport of APR extract (APRE) across human intestinal epithelial Caco-2 cell and in vivo pharmacokinetics in rats after oral administration of APRE. All of the tested coumarins were well or moderately absorbed across Caco-2 monolayers, and could be quickly absorbed into rat blood circulation after oral administration. Columbianetin was the most easily absorbed compound across Caco-2 cell, and also had extremely highest plasma concentration in vivo. Excellent correlation between in vitro absorption across Caco-2 cell monolayers and in vivo pharmacokinetics of coumarins from APRE was well verified. The results provided valuable information for the overall absorption characteristics of the coumarins from APR, as well as for its further studies of in vivo active substances in the further. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Dermal toxicity of 8-methoxypsoralen administered (by gavage) to hairless mice irradiated with long-wave ultraviolet light

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    Langner, A.; Wolska, H.; Marzulli, F.N.; Jablonska, S.; Jarzabek-Chorzelska, M.; Glinski, W.; Pawinska, M.


    Hairless mice were administered various amounts of 8-methoxypsoralen (8-MOP) by gavage, followed by irradiation with ultraviolet light (UVA) two or more times per week for periods ranging from 1 to 12 months. The minimum phototoxic dose was 20 mg/kg body weight by this route of administration and potential for serious organ toxicity in long-term exposures was investigated. No histologic features of cutaneous malignancy were encountered under test conditions which produced prolonged phototoxicity, deep ulceration, cicatrization, and other deformities. Repeated daily gavaged doses of 20 mg psoralen/Kg body weight in conjunction with twice weekly irradiation for 10 min with UVA elicited an erythematous phototoxic reaction, but did not give rise to subsequent skin lesions. 8-MOP in repeated daily gavage doses of 30 mg and 40 mg/kg body weight combined with twice weekly UVA irradiation for 10 min caused severe burning with subsequent scarring, but did not induce malignant tumors in experiments lasting 8 months. No organ toxicity was seen except for toxic liver changes when severe cutaneous burn and pronounced ulcerations were produced. Limited immunologic studies disclosed no abnormalities in this system.

  2. Effect of Imperatorin on the Spontaneous Motor Activity of Rat Isolated Jejunum Strips

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    Marta Mendel


    Full Text Available Imperatorin, a psoralen-type furanocoumarin, is a potent myorelaxant agent acting as a calcium antagonist on vascular smooth muscle. Its effects on other types of smooth muscle remain unknown. Therefore, the aim of this study was to investigate the hypothesized myorelaxant effect of imperatorin on gut motor activity and, possibly, to define the underlying mechanism of action. Imperatorin was made available for pharmacological studies from the fruits of the widely available Angelica officinalis through the application of high-performance countercurrent chromatography (HPCCC. Imperatorin generated reversible relaxation of jejunum strips dose-dependently (1–100 μM. At 25 and 50 μM, imperatorin caused relaxation comparable to the strength of the reaction induced by isoproterenol (Isop at 0.1 μM. The observed response resulted neither from the activation of soluble guanylate cyclase, nor from β-adrenoreceptor involvement, nor from Ca2+-activated potassium channels. Imperatorin relaxed intestine strips precontracted with high potassium concentration, attenuated the force and duration of K+-induced contractions, and modulated the response of jejunum strips to acetylcholine. The results suggest that imperatorin probably interacts with various Ca2+ influx pathways in intestine smooth muscle. The types of some calcium channels involved in the activity of imperatorin will be examined in a subsequent study.

  3. Antimicrobial Photosensitizers: Drug Discovery Under the Spotlight. (United States)

    Yin, Rui; Hamblin, Michael R


    Although photodynamic therapy (PDT) was discovered over a hundred years ago by its ability to destroy microorganisms, it has been developed mainly as a cancer therapy. In recent years, due to the inexorable rise in multi-antibiotic resistant strains of pathogens, PDT is being considered as a versatile antimicrobial approach to which microbial cells will not be able to develop resistance. The goal of this review is to survey the different classes of chemical compounds that have been tested as antimicrobial photosensitizers. Some of these compounds have been known for many years, while others have been rationally designed based on recently discovered structural principles. Tetrapyrrole-based compounds (some of which are approved as cancer therapies) that efficiently generate singlet oxygen are more efficient and broad-spectrum when they bear cationic charges, As the macrocycle structure moves from porphyrins to chlorins to phthalocyanines to bacteriochlorins the long wavelength absorption moves to the near-infrared where tissue penetration is better. Four main types of natural products have been tested: curcumin, riboflavin, hypericin and psoralens. Phenothiazinium dyes, such as methylene blue and toluidine blue, have been tested, and some are clinically approved. A variety of non-phenothiazinium dyes with xanthene, triarylmethane and indocyanine structures have also been tested. New ring structures based on BODIPY, squaraine and fullerene cages can also mediate antimicrobial PDT. Finally the process of photocatalysis using titanium dioxide can also have medical uses. Designing new antimicrobial photosensitizers is likely to keep chemists engaged for a long time to come.

  4. Micose fungóide hipopigmentar com 20 anos de evolução Hypopigmented mycosis fungoides with 20 years evolution

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    Rodrigo Pereira Duquia


    Full Text Available A micose fungóide hipopigmentar é uma forma rara de linfoma cutâneo de células T, em que lesões hipopigmentadas ocorrem na ausência de lesões clássicas dessa neoplasia. Na literatura latino-americana existem apenas três casos; relata-se aqui o caso de um homem negro, de 29 anos com lesões há 20 anos O tratamento é semelhante ao de outros tipos de linfomas T cutâneos, sendo o psoraleno mais ultravioleta A (Puva e a mostarda nitrogenada tópica os mais relatados.Hypopigmented mycosis fungoides is a rare variant of the cutaneous T-cell lymphoma, in which hypopigmented lesions are seen without the classical lesions. Only three cases are found in the Latin-American literature. We report the case of a 29-year-old black male patient, who first presented with these lesions 20 years ago. The treatment is similar to the classical forms of the disease: psoralen plus ultraviolet A (PUVA and topical nitrogen mustard are most often used.

  5. Scanning probe microscope visualization of t-loop assembly by TRF2 in cells (United States)

    Cao, En-Hua; Guo, Xiao-Fe; Wang, Ju-Jun; Qin, Jing-Fen


    Telomeres are essential nucleoprotein structure at the ends of all eukaryotic chromosomes. Our previous work demonstrated that mammalian telomeres were shown to end in a large t-loop structure in vitro and the formation of t-loops was dependent on the presence of TRF2. In this work, the telomere DNA and its complex of TRF2 in HeLa cells has been direct observed in the nanometer resolution regime by atomic force microscopy (AFM) and scanning near-field optical microscopy (SNOM). AFM images showed that the looped structures exited in cell extract containing TRF2, but it disappeared in the protein-deleted samples. When cells were pretreated by UV light plus psoralen, the looped structure could be observed in the protein-deleted samples. SNOM images further demonstrated TRF2 and p53 proteins in cell was bound at the loop junction. Above results suggest that the telomere t-loop structure by TRF2 play a important role in cell-senescence, and might signals p53 protein directly through association with the t-loop junction in cells.

  6. Transmission electron microscopy of the preclinical phase of experimental phytophotodermatitis

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    Hiram Larangeira de Almeida Jr


    Full Text Available OBJECTIVE: To examine the epidermis in induced phytophotodermatitis using transmission electron microscopy in order to detect histologic changes even before lesions are visible by light microscopy. INTRODUCTION: In the first six hours after the experimental induction of phytophotodermatitis, no changes are detectable by light microscopy. Only after 24 hours can keratinocyte necrosis and epidermal vacuolization be detected histologically, and blisters form by 48 hours. METHODS: The dorsum of four adult rats (Rattus norvegicus was manually epilated. After painting the right half of the rat with the peel juice of Tahiti lemon, they were exposed to sunlight for eight minutes under general anesthesia. The left side was used as the control and exposed to sunlight only. Biopsies were performed immediately after photoinduction and one and two hours later, and the tissue was analyzed by transmission electron microscopy. RESULTS: No histological changes were seen on the control side. Immediately after induction, vacuolization in keratinocytes was observed. After one hour, desmosomal changes were also observed in addition to vacuolization. Keratin filaments were not attached to the desmosomal plaque. Free desmosomes and membrane ruptures were also seen. At two hours after induction, similar changes were found, and granular degeneration of keratin was also observed. DISCUSSION: The interaction of sunlight and psoralens generates a photoproduct that damages keratinocyte proteins, leading to keratinocyte necrosis and blister formation. CONCLUSIONS: Transmission electron microscopy can detect vacuolization, lesions of the membrane, and desmosomes in the first two hours after experimental induction of phytophotodermatitis.

  7. Psoralea corylifolia L. Seed Extract Ameliorates Streptozotocin-Induced Diabetes in Mice by Inhibition of Oxidative Stress

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    Eunhui Seo


    Full Text Available Pancreatic beta-cell death is known to be the cause of deficient insulin production in diabetes mellitus. Oxidative stress is one of the major causes of beta-cell death. In this study, we investigated the effects of Psoralea corylifolia L. seed (PCS extract on beta-cell death. Oral administration of PCS extract resulted in a significant improvement of hyperglycemia in streptozotocin-induced diabetic mice. PCS extract treatment improved glucose tolerance and increased serum insulin levels. To study the mechanisms involved, we investigated the effects of PCS extract on H2O2-induced apoptosis in INS-1 cells. Treatment with PCS extract inhibited cell death. PCS extract treatment decreased reactive oxygen species level and activated antioxidative enzymes. Among the major components of PCS extract, psoralen and isopsoralen (coumarins, but not bakuchiol, showed preventive effects against H2O2-induced beta-cell death. These findings indicate that PCS extract may be a potential pharmacological agent to protect against pancreatic beta-cell damage caused by oxidative stress associated with diabetes.

  8. A prospective, open-label study of low-dose total skin electron beam therapy in mycosis fungoides

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    Kamstrup, Maria R; Specht, Lena; Skovgaard, Gunhild L


    PURPOSE: To determine the effect of low-dose (4 Gy) total skin electron beam therapy as a second-line treatment of Stage IB-II mycosis fungoides in a prospective, open-label study. METHODS AND MATERIALS: Ten patients (6 men, 4 women, average age 68.7 years [range, 55-82 years]) with histopatholog......PURPOSE: To determine the effect of low-dose (4 Gy) total skin electron beam therapy as a second-line treatment of Stage IB-II mycosis fungoides in a prospective, open-label study. METHODS AND MATERIALS: Ten patients (6 men, 4 women, average age 68.7 years [range, 55-82 years......]) with histopathologically confirmed mycosis fungoides T2-T4 N0-N1 M0 who did not achieve complete remission or relapsed within 4 months after treatment with psoralen plus ultraviolet-A were included. Treatment consisted of low-dose total skin electron beam therapy administered at a total skin dose of 4 Gy given in 4...... causes and did not complete treatment. Acute side effects included desquamation, xerosis, and erythema of the skin. No severe side effects were observed. CONCLUSION: Low-dose total skin electron beam therapy can induce complete and partial responses in Stage IB-II mycosis fungoides; however, the duration...

  9. Ultraviolet and visible light penetration of epidermis

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    Eggset, G.; Kavli, G.; Volden, G. (Tromsoe Univ. (Norway). Dept. of Dermatology); Krokan, H. (Tromsoe Univ. (Norway). Inst. of Medical Biology)


    Light penetration in untanned skin and skin tanned with UVB (middlewave ultraviolet light) or PUVA (Psoralen photochemotherapy) was compared. Transmission at different wavelengths was measured through sheets of intact epidermis isolated by a suction blister technique. Thick epidermis was collected from a newly formed palmar friction bulla. For these studies a monochromator was used and the range of wavelengths examined was 280-700 nm. The transmission was considerably lower in tanned skin and the difference was most pronounced in the UV range. In the UVB range (290-320 nm), transmission was 13-43% for untanned epidermis, 8-12% for UVB tanned and slightly lower for PUVA tanned epidermis. At wavelengths below 325 nm only a few per cent of light penetrate through thick palmar epidermis. Both UVB and PUVA induce increased melanin content and thickening of the epidermis. Our results indicate that melanin is the most efficient protection against UVA while thickening of epidermis may be as important as the increased melanin content for the protection of living basal cells against the harmful UVB rays.

  10. Primary cutaneous T cell lymphomas: photochemotherapy immunomodulation with analysis of the inflammatory-expansive cellular dynamic. (United States)

    Werber-Bandeira, Luiz; Herdy, Ana Maria; Pagani, Evilmara Adelia; Filgueira, Absalom Lima


    Primary cutaneous T cell lymphomas (CTCLs) are characterized by hyperproliferation of malignant CD4+ T cells with primary localization on the skin. The common characteristics are the migration of the malignant mature T-lymphocytes into the epidermis, with hyperproliferation of malignant CD4+ T cells and epidermotropism. Sézary syndrome (SS) is the leukemic variant. It was established that CTCLs arise from a clonal expansion of CD4+ T cells with an identical rearrangement of the T cell receptor. The purpose of this study was to evaluate the immunomodulation effect of photochemotherapy-A (psoralen plus ultraviolet A (PUVA)). Pre- and post-PUVA punch skin biopsies of nine patients were stained immunohistochemically for CD34+, CD8+, CD7+, CD16+, CD56+, CD1a+, Bcl2+, p53+, CD45RA+, and CD45RO+ cells. The results showed a pre-PUVA cells/mm(2) without significant difference among expansive or reactive cells. Post-PUVA analysis showed a significant decrease in the mean of expansive-reactive cells. PUVA immunomodulated decreasing cellular infiltrate. These findings could contribute to the comprehension of how PUVA acts. We achieved ectoscopic clearance of the lesions, although post-PUVA, there still was a mononuclear pathological infiltrate. This result demonstrates that the PUVA treatment should only be withheld when the histological analysis is normal. © 2013 Wiley Periodicals, Inc.

  11. Analysis of some Italian lemon liquors (limoncello). (United States)

    Andrea, Versari; Nadia, Natali; Teresa, Russo Maria; Andrea, Antonelli


    The chemical composition of several commercial Italian Limoncellos, lemon-peel-based alcoholic beverages, was studied by chromatographic techniques. These methods allowed a rapid monitoring of Limoncello, giving information on quality markers and possible adulteration of the product. Quantitative data for more than 60 compounds are reported. Limoncellos were characterized by the presence of selected volatile (terpenes, aldehydes, alcohols) and nonvolatile compounds (psoralens, coumarins, phenolics, carbohydrates and acids). On the basis of their composition, the samples were grouped by PCA analysis in two sets; the first group showed a composition similar to lemon essential oils, with a high content of b-pinene, myrcene, trans-a-bergamottene, and b-bisabolene, and a low content in neral and geranial. The composition of the second group suggested the occurrence of oxidative phenomena and/or the addition of flavors. The presence of ethyl acetate, acetaldehyde, 2-methyl-1-propanol and glycerol showed that a fermentation probably occurred in the sugar syrup used to dilute the Limoncello after the extraction process.

  12. Stimulation of local immunity and protection in mice by intramuscular immunization with triple- or double-layered rotavirus particles and QS-21. (United States)

    McNeal, M M; Rae, M N; Conner, M E; Ward, R L


    Based on studies in animal models, parenteral immunization has become recognized as a potential vaccination strategy against rotavirus. Using an adult mouse model, the effects of the saponin adjuvant QS-21 on protection against murine rotavirus (strain EDIM) infection was determined following two intramuscular (i.m.) immunizations with purified EDIM particles including triple-layered (tl) infectious particles, tl particles inactivated with psoralen/UV, and double-layered (dl) inactivated particles. All three particles stimulated large serum rotavirus IgG responses and small amounts of serum rotavirus IgA, but undetectable stool rotavirus IgA. Inclusion of QS-21 during immunization increased the serum responses approximately 2- to 10-fold and also stimulated low levels of stool rotavirus IgA. Protection based on reduced shedding of rotavirus following EDIM challenge was significant (P immunized group and was enhanced (P immunization. Mice immunized with either live or inactivated tl particles and QS-21 were almost fully protected. Furthermore, animals inoculated with dl particles and the adjuvant shed significantly (P = .02) less virus following challenge than mice immunized with inactivated tl particles even though the latter induced measurable titers of neutralizing antibody to EDIM. These results demonstrate significant protection against rotavirus following i.m. immunization with both dl and tl EDIM particles which is consistently enhanced with QS-21.

  13. Incidence and Prevalence of Psoriasis in Denmark

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    Alexander Egeberg


    Full Text Available The incidence and temporal trends of psoriasis in Denmark between 2003 and 2012 were examined. There was a female predominance ranging between 50.0% (2007 and 55.4% (2009, and the mean age at time of diagnosis was 47.7–58.7 years. A total of 126,055 patients with psoriasis (prevalence 2.2% were identified. Incidence rates of psoriasis (per 100,000 person years ranged from 107.5 in 2005 to a peak incidence of 199.5 in 2010. Incidence rates were higher for women, and patients aged 60–69 years, respectively. Use of systemic non-biologic agents, i.e. methotrexate, cyclosporine, retinoids, or psoralen plus ultraviolet A (PUVA increased over the study course, and were used in 15.0% of all patients. Biologic agents (efalizumab, etanercept, infliximab, adalimumab, or ustekinumab were utilized in 2.7% of patients. On a national level, incidence of psoriasis fluctuated during the 10-year study course. The relationship between psoriasis incidence and age appeared to be relatively linear, and disease prevalence was comparable to that in other European countries.


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    Herman Cipto


    Full Text Available Lichen planus (LP is an inflammation skin and mucous membrane disease that has characteristic clinical and pathologic features. Typical lesion of LP are small, flat-topped, polygonal, violaceous papules on the skin or reticular, white patch or plaque on mucous membrane. Nail involvement is manifested by longitudinal ridging, thinning and distal splitting and hair involvement is atrophic cicatrical alopesia. There was an increased risk of developing squamous cell carcinoma in the leasion of mucous membrane of oral cavity. Pathology of LP is the prototype of lichenoid interface dermatitis, which shows damage to the basal cell layer and a band-like dental lymphocytic infiltrate in close approximation to the epidermis. Other pathologic findings are compact orthokeratosis, wedge-shape hypergranulosis and irregular acanthosis. The treatment of LP in the main is largely symptomatic like antipruritic for itching. Therapies to suppress immune responses are commonly effective in treating LP, such as corticosteroid and cyclosporine topical or systemic. Other treatment are psoralene ultra violet A (PUVA or steroids.

  15. Implication of the E. coli K12 uvrA and recA genes in the repair of 8-methoxypsoralen-induced mono adducts and crosslinks on plasmid DNA; Implicacion de los genes uvrA de E. coli K12 en la reparacion de monoaductos y entrecruzamien tos inducidos en DNA plasmidico por 8-metoxipso raleno mas luz ultravioleta A

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    Paramio, J.M.; Bauluz, C.; Vidania, R. de


    Genotoxicity of psoralen damages on plasmid DNA has been studied. pBR322 DNA was randomly modified with several concentrations of 8-methoxypsoralen plus 365 nm-UV light. After transformation into E. coli strains (wild-type, uvrA and recA) plasmid survival and mutagenesis were analyzed. To study the influence of the SOS response on plasmid recovery, preirradiation of the cells was performed. In absence of cell preirradiation, crosslinks were not repaired in any strain. Mono adducts were also lethal but in part removed by the excision-repair pathway. Preirradiation of the cells significantly. increased plasmid recovery in recA+ celia. In uvrA- only the mutagenic pathway seemed to be involved in the repair of the damaged DNA. Wild type strain showed the highest increase in plasmid survival, involving the repair of mono adducts and some fraction of crosslinks mainly through an error-free repair pathway. This suggests an enhancement of the excision repair promoted by the induction of SOS functions. (Author) 32 refs.

  16. New tools to study biophysical properties of single molecules and single cells

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    Márcio S. Rocha


    Full Text Available We present a review on two new tools to study biophysical properties of single molecules and single cells. A laser incident through a high numerical aperture microscope objective can trap small dielectric particles near the focus. This arrangement is named optical tweezers. This technique has the advantage to permit manipulation of a single individual object. We use optical tweezers to measure the entropic elasticity of a single DNA molecule and its interaction with the drug Psoralen. Optical tweezers are also used to hold a kidney cell MDCK away from the substrate to allow precise volume measurements of this single cell during an osmotic shock. This procedure allows us to obtain information about membrane water permeability and regulatory volume increase. Defocusing microscopy is a recent technique invented in our laboratory, which allows the observation of transparent objects, by simply defocusing the microscope in a controlled way. Our physical model of a defocused microscope shows that the image contrast observed in this case is proportional to the defocus distance and to the curvature of the transparent object. Defocusing microscopy is very useful to study motility and mechanical properties of cells. We show here the application of defocusing microscopy to measurements of macrophage surface fluctuations and their influence on phagocytosis.Apresentamos uma revisão de duas novas técnicas para estudar propriedades biofísicas de moléculas únicas e células únicas. Um laser incidindo em uma objetiva de microscópio de grande abertura numérica é capaz de aprisionar pequenas partículas dielétricas na região próxima ao foco. Este aparato é chamado de pinça óptica. Esta técnica tem a grande vantagem de permitir a manipulação de um objeto individual. Usamos a pinça óptica para medir a elasticidade entrópica de uma molécula única de DNA em sua interação com o fármaco Psoralen. A pinça óptica também é usada para segurar

  17. Phototherapy for Vitiligo: A Systematic Review and Meta-analysis. (United States)

    Bae, Jung Min; Jung, Han Mi; Hong, Bo Young; Lee, Joo Hee; Choi, Won Joon; Lee, Ji Hae; Kim, Gyong Moon


    References to the expected treatment response to phototherapy would be helpful in the management of vitiligo because phototherapy requires long treatment durations over several months. To estimate the treatment response of vitiligo to phototherapy. A comprehensive database search of MEDLINE, EMBASE, and the Cochrane library from inception to January 26, 2016, was performed for all prospective studies. The main keywords used were vitiligo, phototherapy, psoralen, PUVA, ultraviolet, NBUVB, and narrowband. All prospective studies reporting phototherapy outcome for at least 10 participants with generalized vitiligo were included. Of 319 studies initially identified, the full texts of 141 studies were assessed for eligibility, and 35 were finally included in the analysis. Of these, 29 studies included 1201 patients undergoing narrowband UV-B (NBUVB) phototherapy, and 9 included 227 patients undergoing psoralen-UV-A (PUVA) phototherapy. Two reviewers independently extracted the following data: study design, number and characteristics of the participants, phototherapy protocol, and rate of repigmentation based on the quartile scale. Single-arm meta-analyses were performed for the NBUVB and PUVA groups. Sample size-weighted means were calculated using a random-effects model for the repigmentation rates of the included studies. The primary outcomes were at least mild (≥25%), at least moderate (≥50%), and marked (≥75%) responses on a quartile scale. Response rates were calculated as the number of participants who showed the corresponding repigmentation divided by the number of all participants enrolled in the individual studies. The meta-analysis included 35 unique studies (1428 unique patients). For NBUVB phototherapy, an at least mild response occurred in 62.1% (95% CI, 46.9%-77.3%) of 130 patients in 3 studies at 3 months, 74.2% (95% CI, 68.5%-79.8%) of 232 patients in 11 studies at 6 months, and 75.0% (95% CI, 60.9%-89.2%) of 512 patients in 8 studies at 12 months

  18. Perceptions of U.S. dermatology residency program directors regarding the adequacy of phototherapy training during residency. (United States)

    Goyal, Kavita; Nguyen, Michael O; Reynolds, Rachel V; Mostaghimi, Arash; Joyce, Cara; Cohen, Jeffrey M; Buzney, Elizabeth A


    Phototherapy utilization has declined over the last 20 years despite its efficacy and cost-effectiveness. Adequacy of phototherapy training in residency may be a contributing factor. The purpose of this study was to evaluate perceptions of U.S. dermatology residency program directors (PDs) regarding the effectiveness of their programs' phototherapy training and what constitutes adequate phototherapy education. A questionnaire was sent to PDs to assess phototherapy training within their program; aspects such as dedicated time, exposure to different modalities, and barriers to resident education were surveyed. We assessed the statistical association between these aspects and the perception by PDs that a program's training was adequate. Statistical testing was reported using Fisher's exact tests. A total of 42 PDs responded. Residency training in oral psoralen and ultraviolet A therapy (PUVA), home phototherapy, and excimer laser, respectively, is not provided in 19.0%, 31.0%, and 47.6% of programs. 38.1% of programs provide ≤5 hours of phototherapy training over 3 years of training. 59.5% of PDs cited lack of curriculum time as the most common barrier to phototherapy education. 19.0% of PDs reported completely adequate phototherapy training, which was significantly associated with inclusion of faculty-led didactics, assigned reading, or hands-on clinical training in the curriculum. There is a mismatch between the resources devoted to phototherapy education and the need for dedicated training reported by PDs. Limited time is allocated to phototherapy training during dermatology residency, and a large majority of PDs do not feel that the phototherapy training offered is completely adequate. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  19. Current and emerging therapy for the management of vitiligo

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    Alicia Cecile Borderé


    Full Text Available Alicia Cecile Borderé, Jo Lambert, Nanny van GeelUniversity Hospital of Ghent, Department of Dermatology, Ghent, BelgiumAbstract: Vitiligo is an acquired cutaneous disorder of pigmentation, with an incidence of 0.5% to 2% worldwide. There are three major hypotheses for the pathogenesis of vitiligo that are not exclusive of each other: biochemical/cytotoxic, neural and autoimmune. Recent data provide strong evidence supporting an autoimmune pathogenesis of vitiligo. As vitiligo can have a major effect on quality of life, treatment can be considered and should preferably begin early when the disease is active. Current treatment modalities are directed towards stopping progression of the disease and achieving repigmentation. Therapies include corticosteroids, topical immunomodulators, photo(chemotherapy, surgery, combination therapies and depigmentation of normally pigmented skin. Topical class 3 corticosteroids can be used for localized vitiligo. The use of topical immunomodulators (TIMs in vitiligo seems to be equally effective as topical steroids, especially when used in the face and neck region. In photo(chemotherapy, narrowband ultraviolet-B therapy (NB-UVB seems to be superior to psoralen ultraviolet-A therapy (PUVA and broadband UVB. In surgical techniques, split-thickness grafting and epidermal blister grafting were shown to be effective methods, although the non-cultured epidermal suspension technique has many advantages and seems to be a promising development. Depigmentation therapy can be considered if vitiligo affects more than 60% to 80% of the body. Complementary therapies such as Polypodium leucotomos show promising results in combination with UVB therapy. No causative treatment for vitiligo is currently available. More randomized controlled trials on the treatment of vitiligo are necessary.Keywords: vitiligo, non-surgical treatment, surgical treatment

  20. The Use of Secondary Metabolites Heracleum Sosnowskyi Manden in Agriculture

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    Full Text Available The composition of secondary metabolites Heracleum sosnowskyi include: tannins, essential oils, furanocoumarin and other agent. The greatest interest is the phenolic compounds coumarin furanocoumarin order which is known for their photosensitizing effect. Coumarins are part of hogweed. The greatest number of different kinds of coumarin was found in fruit. Researchers from the Far Eastern Branch of the RAS have been identified series of angelicin furocoumarins (sfondin, angelicin and psoralenum (bergapten, xanthotoxin, oksikumarina umbeliferon furocoumarins. In our research we have identified the influence of active substances hogweed on growth and development of different groups of plants.So experiment with biotest Heracleum sosnowskyi sap (with concentration are 1:1, 1:4, 1:16, and control - H2O stimulates energy germination of pea seeds (Vicia sativa L. - 6%, wheat (Triticum aestivum L. - 1,2%, barley (Hordeum vulgare L. - 5%, and tutsan (Hypéricum perforátum L. - 3,5% at a concentration of 1:16. The same concentration has an inhibitory effect on radish (Raphanus sativus L., chamomile (Matricaria chamomilla L. and oregano (Oríganum vulgáre L..In the experiment on the effect of active substances contained in the soil under hogweed (the control is the soil where wasn’t Heracleum sosnowskyi we observed a stimulatory effect on plant growth and development of wild camomile. So biometric indicators chamomile plants was grown in the A1 horizon (topsoil differ significantly in all indicators of underlying horizons and from controls.So Heracleum sosnowskyi possessing high photosensitizing effect due to content in the bergapten, psoralen, xanthotoxin can be used for medicine along with Ammi majus as the raw material for the preparation of herbal remedies in dermatology. Also in our studies we have shown the use of active substances hogweed in agriculture as regulators growth of some plant species: camomile, wheat, pea, barley, tutsan.

  1. Results of a 5-Week Schedule of Modern Total Skin Electron Beam Radiation Therapy

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    Morris, Stephen Lloyd, E-mail: [St Johns Institute of Dermatology, Guys and St Thomas Hospital, London (United Kingdom); McGovern, Mark; Bayne, Sally; Wain, Mary; Child, Fiona; Whittaker, Sean [St Johns Institute of Dermatology, Guys and St Thomas Hospital, London (United Kingdom)


    Purpose: To report the outcomes of a 5-week schedule of total skin electron beam radiation therapy (TSEB) for mycosis fungoides (MF). Methods: Over 5 years, 41 patients with confirmed MF were treated with a modern TSEB technique delivering 30 Gy in 20 fractions over 5 weeks to the whole skin surface. Data were collected prospectively and entered into the skin tumor unit research database. Skin modified skin weighted assessment tool score data were collected to determine response, duration of response, survival, and toxicity. The outcomes were analyzed according to the patient's stage before TSEB, prognostic factors, and adjuvant treatments. Results: Seventeen patients were stage 1B, 19 were stage IIB, 3 were stage III, and 2 were stage IV. The overall response rate was 95%, with a complete response rate of 51%. Seventy-six percent of patients had relapsed at median follow-up of 18 months. The median time to relapse was 12 months, to systemic therapy was 15 months, and to modified skin weighted assessment tool progression above baseline was 44 months. The complete response rate was 59% in stage IB and 47% in stage IIB patients. The median time to skin relapse was longer in stage IB compared with stage IIB, 18 months versus 9 months. The median time to systemic therapy was longer in stage IB compared with stage IIB, >56 months versus 8 months. The median overall survival was 35 months: >56 months for stage IB, 25 months for stage IIB, 46 months for stage III, and 23.5 months for stage IV. Fifteen patients received adjuvant psoralen + ultraviolet A treatment with no difference seen in the time to relapse. Conclusions: This 5-week schedule of TSEB for MF has a high response rate with comparable duration of response to other regimens. Future studies are needed to find adjuvant and combination treatments to improve the duration of response.

  2. Bath-PUVA therapy improves impaired resting regulatory T cells and increases activated regulatory T cells in psoriasis. (United States)

    Kubo, Ryoji; Muramatsu, Shinnosuke; Sagawa, Yoko; Saito, Chiyo; Kasuya, Saori; Nishioka, Akiko; Nishida, Emi; Yamazaki, Sayuri; Morita, Akimichi


    Bath-psoralen plus ultraviolet light A (PUVA) therapy is an effective, safe, and inexpensive treatment for psoriasis. Psoriasis might be due to an unbalanced ratio of Th17 cells and regulatory T cells (Treg). The Treg functional ratio is significantly lower in patients with psoriasis compared with controls and is inversely correlated with the Psoriasis Area and Severity Index score. We previously reported that bath-PUVA therapy significantly increases the number of Treg and restores Treg function to almost normal in most patients with psoriasis. We examined the effects of bath-PUVA therapy on three distinct Foxp3+ subsets: activated Treg (aTreg), resting Treg (rTreg), and cytokine-secreting non-suppressive T cells. We enrolled 15 patients with psoriasis and 11 healthy controls. We examined aTreg, rTreg, and cytokine-secreting non-suppressive T cells in peripheral blood obtained from the psoriasis patients before and after every fifth bath-PUVA therapy session. Levels of aTreg, which are considered to have the strongest suppressive activity in patients with psoriasis, were significantly increased in the early bath-PUVA therapy sessions, and then diminished. Levels of rTreg were lower in psoriasis patients than in healthy controls, and increased during bath-PUVA therapy. Bath-PUVA therapy induced aTreg and rTreg concomitantly with an improvement in the psoriatic lesions, suggesting a mechanism for the effectiveness of bath-PUVA therapy for psoriasis patients. Copyright © 2017 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.

  3. Isolation and characterization of CYP6B4, a furanocoumarin-inducible cytochrome P450 from a polyphagous caterpillar (Lepidoptera:papilionidae). (United States)

    Hung, C F; Berenbaum, M R; Schuler, M A


    Papilio glaucus (tiger swallowtail) is a generalist that rarely encounters plants containing furanocoumarins yet is constitutively capable of metabolizing low levels of these highly toxic allelochemicals. In larvae of this species, metabolism of linear (xanthotoxin, bergapten), and angular (angelicin, sphondin), furanocoumarins can be induced up to 30-fold by the presence of xanthotoxin in their diet. Degenerate primers corresponding to conserved amino acid sequences in three insect P450s, Musca domestica (CYP6A1), Drosophila melanogaster (CYP6A2) and Papilio polyxenes (CYP6B1), were used to clone xanthotoxin-induced P450 transcripts from P. glaucus larvae by a reverse transcription-polymerase chain reaction (RT-PCR) strategy. Positive clones encoding the highly conserved F--G-R-C-G P450 signature motif were used to isolate a full-length CYP6B4v1 cDNA from a P. glaucus xanthotoxin-induced cDNA library. Sequence comparisons indicate the P. glaucus CYP6B4v1 protein sequence is 63% and 61% identical, respectively, to the P. polyxenes furanocoumarin-inducible CYP6B1v1 and CYP6B3v1 proteins. Northern analysis indicates that CYP6B4 and related transcripts are highly induced in response to xanthotoxin. Baculovirus-mediated expression of the CYP6B4v1 protein in lepidopteran cell lines demonstrates that this P450 isozyme metabolizes isopimpinellin, imperatorin, and bergapten at high rates, xanthotoxin and psoralen at intermediate rates and angelicin, sphondin, and trioxsalen only at very low rates.

  4. Evaluation of PUVA-induced skin side effects in patients referred to the Imam Reza Hospital of Mashhad in 2005-2007

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    Masoud Maleki


    Full Text Available Background: Systemic oral psoralens plus UVA therapy (PUVA is a therapeutic method used with considerable success in many different skin disorders. PUVA therapy causes some cutaneous and noncutaneous side effects and in the present research we deal with cutaneous side effects. Aims: Evaluation of patients to know the different skin side effects of PUVA and their importance. Materials and Methods: All patients referred to the phototherapy unit of Imam Reza Hospital of Mashhad entered the research and skin examination was taken place initially and every 3 months thereafter. Whenever any side effect appeared, it was recorded in the information sheet. Results: One hundred and twenty-eight patients were included in the research, 61 were male between 15 and 75 years and 67 were female between 10 and 61 years of age. Age of female patients at the time of cutaneous side effect appearance was less than male patients. The most common early side effect was pruritus (34.3% and the rarest was telangiectasia (0.7%. One case of late side effect in the form of squamous cell carcinoma was observed in a patient who had received other carcinogenic drugs as well. Complications such as skin dryness, pruritus, erythema and burning sensation occurred at low doses of UVA, while dermatitis, severe limb pain and acne at moderate doses and PUVA lentigines, hypertrichosis and lichenoid lesions appeared at high doses of UVA. Conclusion: Considering the significant therapeutic effects and few serious side effects, PUVA therapy is a suitable and safe method for treatment of certain skin diseases.

  5. Physiological responses of Bituminaria bituminosa to heavy metals. (United States)

    Martínez-Fernández, Domingo; Walker, David J; Romero-Espinar, Pascual; Flores, Pilar; del Río, José Antonio


    Two hydroponic experiments were performed to study the physiological responses to heavy metals (HMs) of two populations of Bituminaria bituminosa (L.) C.H. Stirton (Fabaceae): one ("C2") from a site contaminated by HMs and one from a non-contaminated site ("LA"). In the first, we studied the effects of elevated concentrations of Zn (12 and 61 μM). Population C2 was more tolerant in terms of root and shoot growth at 61 μM Zn, relative to control plants (1 μM Zn). The similar tissue Zn levels of the two populations suggest that C2 is more tolerant of high tissue Zn. Of the parameters measured that could be related to Zn phytotoxicity (micro and macronutrients, root hydraulic activity, water-extractable Zn and organic acids), none could explain totally the superior tolerance of C2. In the second assay, the effects of Cd (4.4 μM), Cu (7.8 μM) and Zn (76 μM) on plant accumulation of the furanocoumarins (FCs) psoralen and angelicin, which function as feeding deterrents and photo-activated toxins, were assessed. For population C2, all three HMs increased the root FC concentrations, while Cd also raised shoot levels. For LA, Cu raised the root concentrations of both FCs. There was a relationship between plant stress, manifested as proline accumulation and disruption of plant water relations, and increased FC accumulation. Higher tissue levels of FCs likely provide greater protection against bacterial or fungal infection and herbivores. Copyright © 2011 Elsevier GmbH. All rights reserved.

  6. Killed but metabolically active Leishmania infantum as a novel whole-cell vaccine for visceral leishmaniasis. (United States)

    Bruhn, Kevin W; Birnbaum, Ron; Haskell, Jacquelyn; Vanchinathan, Veena; Greger, Stephanie; Narayan, Rupa; Chang, Pei-Lin; Tran, Thu Anh; Hickerson, Suzanne M; Beverley, Stephen M; Wilson, Mary E; Craft, Noah


    There are currently no effective vaccines for visceral leishmaniasis, the second most deadly parasitic infection in the world. Here, we describe a novel whole-cell vaccine approach using Leishmania infantum chagasi promastigotes treated with the psoralen compound amotosalen (S-59) and low doses of UV A radiation. This treatment generates permanent, covalent DNA cross-links within parasites and results in Leishmania organisms termed killed but metabolically active (KBMA). In this report, we characterize the in vitro growth characteristics of both KBMA L. major and KBMA L. infantum chagasi. Concentrations of S-59 that generate optimally attenuated parasites were identified. Like live L. infantum chagasi, KBMA L. infantum chagasi parasites were able to initially enter liver cells in vivo after intravenous infection. However, whereas live L. infantum chagasi infection leads to hepatosplenomegaly in mice after 6 months, KBMA L. infantum chagasi parasites were undetectable in the organs of mice at this time point. In vitro, KBMA L. infantum chagasi retained the ability to enter macrophages and induce nitric oxide production. These characteristics of KBMA L. infantum chagasi correlated with the ability to prophylactically protect mice via subcutaneous vaccination at levels similar to vaccination with live, virulent organisms. Splenocytes from mice vaccinated with either live L. infantum chagasi or KBMA L. infantum chagasi displayed similar cytokine patterns in vitro. These results suggest that KBMA technology is a potentially safe and effective novel vaccine strategy against the intracellular protozoan L. infantum chagasi. This approach may represent a new method for whole-cell vaccination against other complex intracellular pathogens.

  7. Phototherapy in atopic dermatitis: a systematic review of the literature. (United States)

    Pérez-Ferriols, A; Aranegui, B; Pujol-Montcusí, J A; Martín-Gorgojo, A; Campos-Domínguez, M; Feltes, R A; Gilaberte, Y; Echeverría-García, B; Alvarez-Pérez, A; García-Doval, I


    Phototherapy is a treatment option for atopic dermatitis recommended by several guidelines. To perform a systematic review of the efficacy of different modalities of phototherapy and photochemotherapy in moderate to severe atopic dermatitis. We considered all randomized clinical trials (RCTs) performed in patients with atopic dermatitis, and accepted all outcome measures. Articles were identified via an online search of the MEDLINE (via Ovid) and Embase databases and the Cochrane Central Register of Controlled Trials. We also searched for clinical trials registered in Current Controlled Trials and in the World Health Organization's International Clinical Trials Registry Platform. Twenty-one RCTs (961 patients) were included in the qualitative analysis. Two of the trials included children and adolescents (32 patients). The efficacy of narrow-band UV-B and UV-A1 phototherapy was similar for the different outcome measures contemplated. Two RCTs assessed the efficacy of psoralen plus UV-A therapy (PUVA). No serious adverse events were described. In general, the publications reviewed were characterized by a high risk of bias and poor reporting of methodology and results. There is evidence for the use of narrow-band UV-B and UV-A1 phototherapy in moderate to severe atopic dermatitis. Evidence supporting the use of PUVA in atopic dermatitis is scarce and there is little information on the use of phototherapy in childhood. For the purpose of future studies, it would be advisable to use comparable criteria and scales for the evaluation of disease severity and patients, to standardize radiation methods, and to establish a minimum follow-up time. Copyright © 2014 Elsevier España, S.L.U. y AEDV. All rights reserved.

  8. Minimal phototoxic dose (MPD) measurements for topical photochemotherapy using a semiautomated MPD tester. (United States)

    Al-Ismail, D; Edwards, C; Al-Ofi, O; Anstey, A V


    The traditional method of assessing minimal phototoxic dose (MPD) prior to photochemotherapy with psoralen-ultraviolet A (PUVA) is inconvenient and cannot directly determine PUVA start doses. A handheld minimal erythema dose UVB tester can be modified by fitting a TL-10 UVA compact fluorescence lamp (CFL). To determine whether MPD testing is possible with a CFL and to calculate a fixed factor to convert observed MPD to PUVA-equivalent MPD. Patients had two sets of MPD tests performed on symmetrical, contralateral sites on the lower back. MPD test results from a panel of PUVA lamps were compared with MPD from the modified handheld tester. Additionally, a questionnaire survey was completed by 43 U.K. phototherapy units to assess routine practice concerning MPD testing prior to PUVA therapy. Thirty-seven patients with psoriasis were recruited. Boston phototypes in the 31 with conclusive MPD reactions were: I, four; II, 11; III, 12; and IV, four. The handheld MPD results were linearly related to the PUVA panel MPD results as follows: PUVA MPD = 0·48 × handheld MPD + 0·17 J cm(-2). The measured PUVA MPD was 0·48 of the handheld MPD, not 0·15 as predicted by the published PUVA action spectrum. The modified MPD tester is a convenient and safe method for PUVA MPD testing, overcoming many problems of the 'traditional method'. The difference between the PUVA and TL-10 lamps was lower than predicted from published studies. This suggests that formal re-evaluation of the erythema action spectrum for PUVA is now needed. © 2013 British Association of Dermatologists.

  9. Immune properties of recombinant vaccinia virus encoding CD154 (CD40L) are determined by expression of virally encoded CD40L and the presence of CD40L protein in viral particles. (United States)

    Bereta, Michal; Bereta, Joanna; Park, Jonas; Medina, Freddy; Kwak, Heesun; Kaufman, Howard L


    Expression of costimulatory molecules by recombinant poxviruses is a promising strategy for enhancing therapeutic vaccines. CD40-CD40L interactions are critical for conditioning dendritic cells (DC) and priming T- and B-cell immunity. We constructed a vaccinia virus expressing murine CD40L (rV-CD40L) and studied its immunomodulatory properties in vitro. Direct DC infection with control vaccinia or psoralen/UV-inactivated rV-CD40L stimulated high levels of interleukin 12 (IL-12) release. However, replication-competent rV-CD40L did not stimulate IL-12 under similar conditions. We observed a high level of CD40L protein on purified viral particles and demonstrated that induction of IL-12 by nonreplicating rV-CD40L was blocked by anti-CD40 antibodies suggesting that functional CD40L on viral particles contributed to alterations in IL-12 synthesis. Since cross-presentation of tumor-associated antigens by DC is augmented by viral infection of tumor cells, we infected MC38 murine colon carcinoma cells with rV-CD40L. Infected cells stimulated IL-12 secretion by DC and proliferation of B cells and DX5(+) (NK/NKT) cells through direct CD40-CD40L interaction. A subpopulation of NKT cells expressing CD40 (NK1.1(+), CD3(lo)) appeared to be a major effector population responding to MC38/rV-CD40L. These results highlight the complex immune regulatory effects of rV-CD40L defined by the cumulative effects of CD40L expression, presence of CD40L protein in viral particles, and the replication potential of the virus.

  10. Management of hair loss diseases

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    Manabu Ohyama


    Full Text Available The treatment of hair loss diseases is sometimes difficult because of insufficient efficacy and limited options. However, recent advances in understanding of the pathophysiology and development of new remedies have improved the treatment of refractory hair loss conditions. In this article, an update on the management of hair loss diseases is provided, especially focusing on recently reported therapeutic approaches for alopecia areata (AA. An accurate diagnosis is indispensable to optimize treatment. Dry dermoscopy represents new diagnostic techniques, which could enable the differentiation of barely indistinguishable alopecias, e.g. AA and trichotillomania. An organized scalp biopsy adopting both vertical and transverse sectioning approaches also provides a deep insight into the pathophysiology of ongoing alopecias. Among various treatments for AA, intraregional corticosteroid and contact immunotherapy have been recognized as first-line therapies. However, some AA cases are refractory to both treatments. Recent studies have demonstrated the efficacy of pulse corticosteroid therapy or the combination of oral psoralen ultraviolet A therapy and systemic corticosteroids for severe AA. Previous clinical observations have suggested the potential role of antihistamines as supportive medications for AA. Experimental evaluation using AA model mice further supports their effectiveness in AA treatment. Finasteride opens up new possibilities for the treatment of androgenetic alopecia. For androgenetic alopecia patients refractory to finasteride, the combination of finasteride with topical minoxidil or the administration of dutasteride, another 5 alpha-reductase inhibitor, may provide better outcomes. Scarring alopecia is the most difficult form of hair loss disorder to treat. The bulge stem cell area is destroyed by unnecessary immune reactions with resultant permanent loss of hair follicle structures in scarring alopecia. Currently, treatment options for

  11. Photogenotoxicity of skin phototumorigenic fluoroquinolone antibiotics detected using the comet assay. (United States)

    Reavy, H J; Traynor, N J; Gibbs, N K


    The fluoroquinolone (FQ) antibiotics photosensitize human skin to solar UV radiation and are reported to photosensitize tumor formation in mouse skin. As tumor initiation will not occur without genotoxic insult, we examined the potential of ciprofloxacin, lomefloxacin, fleroxacin, BAYy3118 (a recently developed monofluorinated quinolone) and a nalidixic acid to photosensitize DNA damage in V79 hamster fibroblasts in vitro. Cells were exposed to 37.5 kJ/m2 UVA (320-400 nm; glass filtered Sylvania psoralen + UVA (PUVA) tubes; calibrated Waldmann radiometer) at 4 degrees C in the presence of FQ and immediately afterwards embedded in agarose, lysed and placed in an electrophoretic field at pH 12. Under these denaturing conditions, the presence of DNA single-strand breaks (SSB), alkali-labile sites (ALS) and double-strand breaks (DSB) can be visualized as DNA migrating away from the nucleus (characteristic "comet" appearance) after staining with a specific fluorochrome. At FQ concentrations that induced minimal loss of cell viability (neutral red uptake assay) the compounds tested induced comets with a rank order of BAYy3118 > norfloxacin > ciprofloxacin > lomefloxacin > fleroxacin > nalidixic acid. If cells were incubated after treatment for 1 h at 37 degrees C, the comet score decreased, suggesting efficient removal of SSB/ALS/DSB. Addition of the DNA polymerase(alpha) inhibitor, aphidicolin, to cells treated with either ciprofloxacin alone or ciprofloxacin + UVA resulted in an accumulation of SSB due to the endo/exonuclease steps of excision repair. We have demonstrated that the FQ are photogenotoxic in mammalian cells but the FQ-photosensitized SSB are efficiently repaired. Preliminary evidence that ciprofloxacin photosensitizes the formation of DNA lesions warranting excision repair may indicate production of more mutagenic lesions.

  12. Coumarins from Angelica decursiva inhibit α-glucosidase activity and protein tyrosine phosphatase 1B. (United States)

    Ali, Md Yousof; Jannat, Susoma; Jung, Hyun Ah; Jeong, Hyong Oh; Chung, Hae Young; Choi, Jae Sue


    In the present study, we investigated the anti-diabetic potential of six natural coumarins, 4-hydroxy Pd-C-III (1), 4'-methoxy Pd-C-I (2), decursinol (3), decursidin (4), umbelliferone 6-carboxylic acid (5), and 2'-isopropyl psoralene (6) isolated from Angelica decursiva and evaluated their inhibitory activities against protein tyrosine phosphatase 1B (PTP1B), α-glucosidase, and ONOO(-)-mediated protein tyrosine nitration. Coumarins 1-6 showed potent PTP1B and α-glucosidase inhibitory activities with ranges of IC50 values of 5.39-58.90 μM and 65.29-172.10 μM, respectively. In the kinetic study for PTP1B enzyme inhibition, compounds 1, 5, and 6 were competitive, whereas 2 and 4 showed mixed type, and 3 displayed noncompetitive type inhibition. For α-glucosidase enzyme inhibition, compounds 1 and 3 exhibited good mixed-type, while 2, 5, and 6 showed noncompetitive and 4 displayed competitive type inhibition. Furthermore, these coumarins also effectively suppressed ONOO(-)-mediated tyrosine nitration in a dose-dependent manner. To further investigate PTP1B inhibition, we generated a 3D structure of PTP1B using Autodock 4.2 and simulated the binding of compounds 1-6. Docking simulations showed that different residues of PTP1B interacted with different functional groups of compounds 1-6 through hydrogen and hydrophobic interactions. In addition, the binding energies of compounds 1-6 were negative, suggesting that hydrogen bonding may stabilize the open form of the enzyme and potentiate tight binding of the active site of PTP1B, thereby resulting in more effective PTP1B inhibition. These results demonstrate that the whole plant of A. decursiva and its coumarins are useful as potential functional food ingredients for the prevention and treatment of type 2 diabetes. Copyright © 2016. Published by Elsevier Ireland Ltd.

  13. 4,6,4'-trimethylangelicin shows high anti-proliferative activity on DU145 cells under both UVA and blue light. (United States)

    Miolo, G; Sturaro, G; Cigolini, G; Menilli, L; Tasso, A; Zago, I; Conconi, M T


    Furocoumarins (psoralens and angelicins) have been already used under ultraviolet A light (UVA) for the treatment of skin diseases and cutaneous T-cell lymphoma. Besides their high anti-proliferative activity, some severe long-term side effects have been observed, for example genotoxicity and mutagenicity, likely strictly related to the formation of crosslinks. It has been demonstrated that blue light (BL) activation of 8-methoxypsoralen, an FDA-approved drug, leads to less mutagenic monoadducts in the DNA. So far, in this work the less toxic and more penetrating BL is proposed to activate 4,6,4'-trimethylangelicin (TMA), an already known UVA photoactivatable compound. Photocleavage, crosslink formation and oxidative damage were detected in pBR322 plasmid DNA treated with 300.0 μmol/L TMA activated with various exposures of BL. Anti-proliferative activity, reactive oxygen species (ROS) formation and activation status of some signalling pathways involved in cell growth and apoptosis were verified on DU145 cells treated with 5.0 μmol/L TMA plus 2.0 J/cm2 of BL. Under BL-TMA, no mutagenic crosslinks, no photocleavage and neither photooxidative lesions were detected on isolated plasmid DNA. TMA showed high anti-proliferative activity on DU145 cells through induction of apoptosis. Besides ROS generation, the proapoptotic effect seemed to be related to activation of p38 and inhibition of p44/42 phosphorylation. Interestingly, the decrease in nuclear β-catenin was coupled with a significant dropping of CD44-positive cells. Overall, our results indicate that TMA can be activated by BL and may be considered for targeted phototherapy of prostate cancer lesions. © 2018 John Wiley & Sons Ltd.

  14. Photo(chemotherapy reduces circulating Th17 cells and restores circulating regulatory T cells in psoriasis.

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    Takuya Furuhashi

    Full Text Available BACKGROUND: Photo(chemotherapy is widely used to treat psoriasis, the pathogenesis of which might be caused by an imbalance of Th17 cells/regulatory T cells (Treg. In the present study, we evaluated the effects of photo(chemotherapy on the Th17/Treg balance and Treg function. METHODS: Peripheral blood was obtained from psoriasis patients treated with bath-psoralen ultraviolet A (UVA, n = 50 or narrowband ultraviolet B (UVB, n = 18, and age-matched healthy volunteers (n = 20. CD3(+CD4(+IL-17A(+ or CD4(+CD25(+Foxp3(+cells were analyzed to estimate Th17 or Treg number by fluorescence-activated cell sorting. Moreover, CD4(+ CD25(- T cells from patients treated with PUVA(n = 14 were incubated in CFSE and activated with or without CD4(+ CD25(+T cells, and the suppressive function of CD4(+ CD25(+T cells were analyzed. RESULTS: Photo(chemotherapy significantly reduced Th17 levels from 5.66 ± 3.15% to 2.96 ± 2.89% in patients with increased Th17 (Th17/CD4>3.01% [mean+SD of controls]. In contrast, photo(chemotherapy significantly increased Treg levels from 2.77 ± 0.75 to 3.40 ± 1.88% in patients with less than 4.07% Treg level, defined as the mean of controls. Furthermore, while Treg suppressed the CD4(+CD25(- T cell proliferation to a greater extent in controls (Treg Functional Ratio 94.4 ± 4.28% than in patients (70.3±25.1%, PUVA significantly increased Treg Functional Ratio to 88.1 ± 6.47%. Th17 levels in severe patients (>30 PASI were significantly higher as compared to controls. Th17 levels that were left after treatment in the patients not achieving PASI 50 (3.78 ± 4.18% were significantly higher than those in the patients achieving PASI 75 (1.83±1.87%. Treg levels in patients achieving PASI 90 (4.89 ± 1.70% were significantly higher than those in the patients not achieving PASI 90 (3.90 ± 1.66%. Treg levels prior to treatment with Th17 high decreased group (5.16 ± 2.20% was significantly higher than that with Th17 high increased group

  15. Clinical importance of Demodex folliculorum in patients receiving phototherapy. (United States)

    Kulac, Mustafa; Ciftci, Ihsan Hakki; Karaca, Semsettin; Cetinkaya, Zafer


    Patients with immunodeficiency are prone to infestation with Demodex folliculorum mites. Ultraviolet (UV) radiation can lead to immunosuppression and sebaceous gland hyperplasia. Although some cases of demodicidosis related to UV radiation exposure have been reported, no studies have been performed on the incidence of D. folliculorum and its clinical characteristics in patients receiving phototherapy. Objective To investigate the effects of phototherapy on the density of D. folliculorum infestation and its clinical characteristics. This was a cross-sectional study. Forty-five patients receiving phototherapy and 43 age- and sex-matched healthy controls were enrolled to the study. The sociodemographic characteristics, occupational information, and skin types (2, 3, 4, or 5) of both patients and controls were carefully recorded. The dermatologic diseases requiring phototherapy, type and number of phototherapy treatments, and cumulative UV doses of all patients were noted. The clinical findings that may relate to demodicidosis were recorded. Standardized skin surface biopsies were taken from three anatomic regions (forehead, cheek, and nasal dorsum) and suspected lesions; five or more D. folliculorum mites per square centimeter of skin was defined as demodicidosis. Twelve (26.7%) patients received psoralen plus UV-A (PUVA) and 33 (73.3%) received narrow-band UV-B. Demodicidosis was detected in 13 (28.9%) patients and three (7%) controls. The difference in the demodicidosis rate between patients and controls was statistically significant (P = 0.01). In eight of the 13 patients (61.5%) with demodicidosis, clinical demodicidosis was present. Demodicidosis was present in seven of the 12 patients (58.3%) receiving PUVA and in six of the 33 patients (18.2%) receiving narrow-band UV-B. The difference in demodicidosis rates between patients receiving PUVA and those receiving narrow-band UV-B was statistically significant (P = 0.02). A statistically significant difference was


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    Kavitha S. B


    alternative such as steroids/PUVA as initial therapy in paediatric age where psoralens and steroids should not be initiated because of long-term risks associated with prolonged usage of them as adjunctive therapy to steroids or tacrolimus so as to reduce the dose and duration of steroids or tacrolimus for increasing the overall safety to the child.

  17. Epidermal hydrogen peroxide is not increased in lesional and non-lesional skin of vitiligo. (United States)

    Zailaie, Mohammad Z


    It is widely believed that the loss of the epidermal melanocytes in vitiligo is basically due to excessive oxidative stress. Previous research work described abnormal elevation of the absolute concentration of the epidermal hydrogen peroxide (H2O2) in lesional and non-lesional skin of vitiligo. Based on this finding, our primary research objective was to use this feature as a screening marker in individuals at a great risk of developing vitiligo. Ninety-six patients of non-segmental vitiligo (NSV) of varying durations, skin phototypes, and treatment modalities (psoralen UVA-, narrow band UVB-treated) were recruited for this study. Raman spectroscopic measurements, using an external probehead, of the lesional and non-lesional skin were obtained, and the resulting spectra were analyzed using the Opus software package of the MultiRam spectrometer and the intensity of the peak at 875 cm-1 that represents the absolute concentration of H2O2 was calculated. Contrary to previous reports, in patients of skin phototype IV, the absolute concentrations of H2O2 in non-lesional and lesional NSV of all groups were non-significantly decreased compared to normal control. In patients of NSV of skin phototype V, the decrease in the absolute concentrations of H2O2 was not significant in the untreated group, and a slight non-significant increase in the NBUVB-treated group was noted. However, in the PUVA-treated group, the non-lesional skin demonstrated significant increase in the absolute concentration of H2O2, whereas the lesional skin showed only a slight non-significant increase compared to normal control. In NSV patients of skin phototype VI who were previously treated with PUVA, the non-lesional skin showed a slight non-significant increase in the absolute concentration of H2O2; however, the lesional skin showed a marked significant decrease compared to normal control and the non-lesional skin. Thereof, one can conclude that the epidermal H2O2 is not increased in NSV as

  18. Experience of Phototherapy in Dermatological Praxis in Complex Therapy of Psoriasis Patients

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    Hartmane Ilona


    Full Text Available Psoriasis is a chronic relapsing skin disease presenting with erythematous and papulous lesions with infiltration and extensive desquamation on the skin surface. It is a genetically determined, multifactorial dermatosis where genetic, immune, and environmental factors play significant roles in its development. In Latvia in treatment of different forms of extensively spreading psoriasis, PUVA (psoralen and ultraviolet A light therapy, a combined method, is administered, applying long-wave UVA radiation with wavelength 320–400 nm in combination with photosensibilisator 8-metoxypsoralen and medium wave length UVB radiation narrow-band phototherapy — 311 nm using specialised TL-01 lamps. The aim of our clinical investigation was to determine the efficacy of narrow-band phototherapy (UVB 311 nm in the complex treatment of patients with different severity of extensive psoriatic lesions treated in the Clinical Centre of Skin and Sexually Transmitted Diseases. Cases of clinical data of 260 patients with widely spread psoriasis were analysed. In the Group 1 (n = 102 receiving narrow-band UVB therapy, the mean and cumulative UVB dosage was 1.8 ± 0.6 and 21.5 ± 3.8 J/cm2, respectively, whereas in Group 2 (n = 91 it was 2.2 ± 0.1 and 27.7 ± 8.0 J/cm2. To obtain clinical recovery, 18 to 30 procedures were necessary (average 22 ± 4.1 with total irradiation dose received 110 ± 4.6 J/cm2. In 67 patients of the control group, PUVA therapy was administered, and positive therapeutic efficacy was observed in all patients. Clinical recovery was obtained in 86.2% in patients of the Group 1, in 82.4% — of Group 2, and in 80% — in 67 patients of the control group. Narrow-band (311 nm UVB phototherapy is currently one of the leading pathogenetical methods of treatment of patients with widespread psoriasis. It has high efficacy, good tolerability, does not have severe side effects and restrictions in use, in comparison with traditional PUVA therapy.

  19. Protective effect of superoxide dismutase against hair graying in a mouse model. (United States)

    Emerit, I; Filipe, P; Freitas, J; Vassy, J


    Oxygen free radicals play a role in the aging process, and the protective effect of various antioxidants has been intensively studied, in particular for cutaneous aging. Besides hereditary factors, free radical-mediated damage to melanocytes of the hair follicle has been considered as a mechanism for aging of the hair. It was the aim of this study to evaluate the role of photosensitization reactions for hair graying and to demonstrate potential protective effects of superoxide dismutase (SOD). Mice with black hair were depilated with the fingertips on a surface of 6 x 2.5 cm on both sides of the dorsum. The right side received five applications of a SOD-containing gel before exposure to psoralen (concentration 0.5 mg/mL) plus UV-A (365 nm, 4 J/cm2). The left side was pretreated in the same way with a gel free of SOD. When the hair started growing again, the SOD-protected side was covered with black hair, whereas the hair on the vehicle-treated side was gray or white in 27 of the 30 animals studied. The 0.01% SOD concentration was as protective as the 0.1% concentration. Heat-inactivated SOD, applied in another five animals, was not protective. Using fluorescent labeling of the SOD with fluorescein isothiocyanate, epifluorescence microscopy and digital imaging processing, we show that SOD applied to the skin surface penetrates through the follicular appendages, as well as through the unbroken stratum corneum. Our findings suggest that superoxide radicals, generated by interaction of UV-A light with the sensitizer, initiated the formation of secondary products with well-known DNA-damaging effects, such as lipid peroxidation products and tumor necrosis factor alpha. SOD prevented the damage to melanocyte DNA by dismutating superoxide. Photosensitization may be another mechanism for hair graying, which can be influenced by antioxidants. Given the large number of exogenous and endogenous sensitizers, this mechanism deserves further study for human hair graying.

  20. Efficacy and safety of etanercept in psoriasis after switching from other treatments: an observational study. (United States)

    Mazzotta, Annamaria; Esposito, Maria; Costanzo, Antonio; Chimenti, Sergio


    Since targeted biologic treatments have been introduced for the treatment of plaque-type psoriasis and psoriatic arthritis, switching between different medications has become necessary in selected patients, particularly after treatment failures. To evaluate the efficacy and safety of etanercept treatment in adult patients with psoriasis after failure to respond to other previous therapies. In particular, the differences in efficacy profiles after switching from traditional (cyclosporine [ciclosporin], methotrexate, retinoids, fumaric acid esters, psoralen plus UVA therapy, corticosteroids) or biologic (infliximab, efalizumab) treatments were analyzed. The study included 124 patients affected by plaque-type psoriasis who received etanercept administered subcutaneously at a dosage of 50 mg twice weekly for 12 weeks, followed by 25 mg twice weekly for an additional 12 weeks, and 110 patients affected by psoriatic arthritis who were treated with etanercept 25 mg twice weekly in a continuous regimen, after a 12-week period of treatment with etanercept 50 mg twice weekly. Efficacy results were consistent in both groups of patients (plaque-type psoriasis and psoriatic arthritis), as expressed by the percentage of patients who achieved Psoriasis Area and Severity Index (PASI) 50 and PASI 75 scores. Among psoriatic arthritis patients, the mean pain Visual Analog Scale (VAS) score showed a substantial reduction during the treatment course, from 67.2 at week 0 to 15.8 at week 24. After 24 weeks, among patients with plaque-type psoriasis who had not previously received biologic therapies, 89.9% of patients achieved PASI 50 and 75.3% achieved PASI 75, while among patients who had received biologic therapies, 69.6% of patients achieved PASI 50 and 65.2% achieved PASI 75. In addition, 92.3% of patients with psoriatic arthritis who had not previously received biologic therapies achieved PASI 50 and 73.8% achieved PASI 75, while among patients who had received biologic therapies

  1. Effectiveness of Subcutaneous Methotrexate in Chronic Plaque Psoriasis. (United States)

    Yesudian, Paul Devakar; Leman, Joyce; Balasubramaniam, Periasamy; Macfarlane, Andy W; Al-Niaimi, Firas; Griffiths, Christopher E M; Burden, Arthur David; Warren, Richard B


    Oral methotrexate (MTX) has been a first line systemic agent in the treatment of chronic plaque psoriasis (CPP) for more than 50 years. Parenteral MTX, administered as a subcutaneous (SC) injection has gained favour in recent years. The effectiveness of SC MTX has been proven in rheumatological conditions but there has been no assessment of its role in CPP. We retrospectively reviewed case notes of 85 patients prescribed SC MTX for psoriasis in three dermatology centres in the UK (Betsi Cadwaladr University Health Board, Western Infirmary, Glasgow, and Salford Royal NHS Foundation Trust). Audit department approval was sought and granted. A total of 85 patients (44 male; 41 female; age range 14 - 78 years, mean 44 years; 79 Caucasian, 6 Asian) with CPP were identified. The average duration of psoriasis was 19 years [range 3 - 60 years]. Co-morbidities included depression, diabetes mellitus, hypertension, epilepsy, obesity, ischaemic heart disease, and hyperlipidaemia; 29 patients had no associated co-morbidities. Psoriatic arthritis was noted in 18 patients. Previous treatments included phototherapy (both narrow band ultraviolet B [TLO1] and psoralen and ultraviolet A [PUVA])(n=60), oral MTX (n=82), ciclosporin (n=37), acitretin (n=19), fumaric acid esters (n=20), hydroxycarbamide (n=6), mycophenolate mofetil (n=2), and repeated in-patient admissions (n=2). Oral MTX was stopped due to nausea (n=43), ineffectiveness (n=13) or partial response (n=11), headache (n=3), increased liver enzymes (n=2), and lethargy (n=2). The median number of systemic agents used prior to SC MTX was 3 (mean 2.65, range 1 to 6 agents). The weekly dose of SC MTX varied between 7.5mg to 30 mg (mean 18.5mg, median 20mg) and had been used for 2 months to 67 months (mean 14 months; median 9 months). Folic acid supplementation was used in every patient. The patients were reviewed between 6 weeks to 3 months once treatment was fully established. Using a pre-determined "adjective list" (where

  2. Morfoanatomia e histoquímica de Brosimum gaudichaudii Trécul (Moraceae Morphoanatomy and histochemistry of the Brosimum gaudichaudii Trécul (Moraceae

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    Ezilda Jacomassi


    Full Text Available Brosimum gaudichaudii, conhecida como mamacadela, é espécie comum nos cerrados, com grande valor na medicina popular e na indústria de medicamentos. A casca da raiz e do caule é empregada no tratamento do vitiligo, sendo o bergapteno e psoraleno, duas furanocumarinas, responsáveis pela ação farmacológica. Neste trabalho são apresentadas a caracterização morfoanatômica e a histoquímica de órgãos vegetativos de B. gaudichaudii, com informações sobre o embrião e o desenvolvimento pós-seminal. Amostras foram processadas segundo técnicas usuais em anatomia e ultra-estrutura vegetal. O embrião é do tipo total e invaginado, a plântula cripto-hipogéia e o sistema radicular do tirodendro é composto por raiz primária pouco desenvolvida em relação às raízes laterais. Plantas adultas apresentam raízes gemíferas longas, plagiotrópicas originando ramos caulinares aéreos; tais raízes possuem periderme com súber desenvolvido e floema secundário abundante em relação ao xilema, sendo facilmente destacados quando manipulados. O caule jovem e a folha apresentam epiderme unisseriada, grande densidade de tricomas tectores unicelulares e tricomas glandulares. Laticíferos não-articulados ramificados ocorrem em todos os órgãos, podendo invadir os elementos de vaso. Idioblastos fenólicos ocorrem em abundância em todos tecidos e órgãos analisados. O valor das características anatômicas observadas no controle de qualidade da droga é discutido.Brosimum gaudichaudii known as "mamacadela" is a common cerrado species with a great importance on medicine folk and in the pharmaceutical industry. The bark of the root and stem is used on the treatment of "vitiligo" being the bergapten and psoralen two furocoumarins responsible for pharmacological activity. This work presents the morphoanatomical and histochemical characterization of the vegetative organs of B. gaudichaudii, with information about its embryo and post

  3. Phenolic Compounds of Fresh Roots and Rhizomes of Garden Angelica Angelica Archangelica L.

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    Full Text Available Because in the process of drying and storage of plant materials is a change of genuine compounds, we studied the content of phenolic compounds in freshly drawn angelica roots and rhizomes of the drug Angelica archangelica L. samples had been taken on the territory of the Bashkir State University experimental plot a few days before soil freezing. Fresh rhizomes and roots were ground at room temperature and poured with 96% ethanol. The first portion of the extract containing the glycosides was kept at +5 ° C to precipitate the polysaccharides, and the residual plant material was extracted repeatedly during the day of 96% ethanol to isolate coumarins.After two hours, the first portion of the extract containing glycosides was frozen in a refrigerator, and the remaining part was re-extracted by 96% ethanol for coumarin isolation next day. The second portion representing 80% of the ethanol extract was evaporated to a "tar" state under vacuum at room temperature. Then it was dissolved in CHCl3 and placed in the refrigerator. After a day of keeping the chloroform extract at -10°C, its separation into two fractions was observed: a yellow-orange top and a poor-colored bottom.After separation a gas chromatography-mass spectrometer Thermo Finnigan, 800-Finnigan chromatograph and mass spectrometer of high resolution MAT-95XP "Delta" with “Data System” processing were used.In the alcohol fraction (50% 6.45% there were found catechol, methylcatechol 0.61%, 4.23% 2-methoxy-4-vinylphenol, xanthyletine 0.78%, 4.21% osthol, 10.47% 2,2-dimethyl-3,4-dihydro-2H,5H-pyrano[2,3-b]chromen-5-one, 1.35% kvannin, isoangenomalin 0.37%, 0.8% psoralen and 1.32% prangenin.In the colored fractions 0.41 % bergaptol, 0.41 % marmesin, 0.20 % oroselonе, and 0.07 % neobyakangelikol have been found. High levels of ethyl α-D-glucopyranoside amounting to 35 % were observed in this fraction as well. furocoumarin glycosides are entirely possible to be hydrolysed during the

  4. Knowledge about Ultraviolet Radiation Hazards and Tanning Behavior of Cosmetology and Medical Students. (United States)

    Zuba, Ewelina Bogumiła; Francuzik, Wojciech; Malicki, Przemysław; Osmola-Mańkowska, Agnieszka; Jenerowicz, Dorota


    (SPF), and tanorexia. The students were additionally asked to mention possible side effects of solar radiation and contraindications to sunbeds and drugs, which may induce photosensitivity. Statistical analysis was performed using The R Project for Statistical Computing. Chi-squared test was used to compare both sun-risk knowledge and tanning behaviors between medical and cosmetology students. Pcosmetology students more frequently gave a correct definition. In the group of students who stated they knew the definition of skin phototype, medical students were significantly more frequently wrong when we asked them to explain the term in their own words. Cosmetology students correctly answered significantly more knowledge checking questions (Table 2). When we asked students to list photosensitizing agents, students of the cosmetology program gave twice as many correct answers per respondent as students of the medicine program (see Table 3). Cosmetology students more frequently listed retinoids, while medical students listed tetracyclines as the main photosensitizing drug. The most common answer in the cosmetology group was the herb of Hypericum perforatum, although it is not considered a drug. Psoralens were identified by only 4 medical students as a possible cause of phototoxicity. When students were asked to list adverse effects of sunbathing, we specifically looked for three responses (see Table 4). Cosmetology students listed those answers significantly more often than medical students. Students of the cosmetology program gave significantly more correct answers when asked to list contraindications for sunbathing. While medical students reported mainly pregnancy (as a contraindication for most medical procedures), cosmetology students reported history of skin cancer as the most frequent answer (Table 5). Cosmetology students (89.04%) stated they visited a tanning salon more often than medical students (46.55%) (Pcosmetology and medical female students. Cosmetology