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Pesquisa : B01.050.150.900.649.573.600 [Categoria DeCS]
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[PMID]:28040568
[Au] Autor:Botero A; Keatley S; Peacock C; Thompson RC
[Ad] Endereço:School of Veterinary and Life Sciences, Murdoch University, South Street, Murdoch, WA 6150, Australia. Electronic address: L.Boterogomez@murdoch.edu.au.
[Ti] Título:In vitro drug susceptibility of two strains of the wildlife trypanosome, Trypanosoma copemani: A comparison with Trypanosoma cruzi.
[So] Source:Int J Parasitol Drugs Drug Resist;7(1):34-41, 2017 Apr.
[Is] ISSN:2211-3207
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Trypanosomes are blood protozoan parasites that are capable of producing illness in the vertebrate host. Within Australia, several native Trypanosoma species have been described infecting wildlife. However, only Trypanosoma copemani has been associated with pathological lesions in wildlife hosts and more recently has been associated with the drastic decline of the critically endangered woylie (Bettongia penicillata). The impact that some trypanosomes have on the health of the vertebrate host has led to the development of numerous drug compounds that could inhibit the growth or kill the parasite. This study investigated and compared the in vitro susceptibility of two strains of T. copemani (G1 and G2) and one strain of Trypanosoma cruzi (10R26) against drugs that are known to show trypanocidal activity (benznidazole, posaconazole, miltefosine and melarsoprol) and against four lead compounds, two fenarimols and two pyridine derivatives (EPL-BS1937, EPL-BS2391, EPL-BS0967, and EPL-BS1246), that have been developed primarily against T.cruzi. The in vitro cytotoxicity of all drugs against L6 rat myoblast cells was also assessed. Results showed that both strains of T. copemani were more susceptible to all drugs and lead compounds than T. cruzi, with all IC50 values in the low and sub-µM range for both species. Melarsoprol and miltefosine exhibited the highest drug activity against both T. copemani and T. cruzi, but they also showed the highest toxicity in L6 cells. Interestingly, both fenarimol and pyridine derivative compounds were more active against T. copemani and T. cruzi than the reference drugs benznidazole and posaconazole. T. copemani strains exhibited differences in susceptibility to all drugs demonstrating once again considerable differences in their biological behaviour.
[Mh] Termos MeSH primário: Animais Selvagens/parasitologia
Tripanossomicidas/farmacologia
Trypanosoma cruzi/efeitos dos fármacos
Trypanosoma/efeitos dos fármacos
[Mh] Termos MeSH secundário: Animais
Austrália
Linhagem Celular
Descoberta de Drogas
Ensaios de Triagem em Larga Escala
Concentração Inibidora 50
Melarsoprol/farmacologia
Melarsoprol/toxicidade
Nitroimidazóis/farmacologia
Fosforilcolina/análogos & derivados
Fosforilcolina/farmacologia
Fosforilcolina/toxicidade
Potoroidae/parasitologia
Pirimidinas/farmacologia
Triazóis/farmacologia
Trypanosoma/isolamento & purificação
Trypanosoma cruzi/isolamento & purificação
[Pt] Tipo de publicação:COMPARATIVE STUDY; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Nitroimidazoles); 0 (Pyrimidines); 0 (Triazoles); 0 (Trypanocidal Agents); 107-73-3 (Phosphorylcholine); 53EY29W7EC (miltefosine); 6TK1G07BHZ (posaconazole); YC42NRJ1ZD (benzonidazole); ZF3786Q2E8 (Melarsoprol)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171024
[Lr] Data última revisão:
171024
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170102
[St] Status:MEDLINE


  2 / 114 MEDLINE  
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[PMID]:27561173
[Au] Autor:Botero A; Cooper C; Thompson CK; Clode PL; Rose K; Thompson RC
[Ad] Endereço:School of Veterinary and Biomedical Sciences, Murdoch University, Murdoch, Western Australia 2009, Australia.
[Ti] Título:Morphological and Phylogenetic Description of Trypanosoma noyesi sp. nov.: An Australian Wildlife Trypanosome within the T. cruzi Clade.
[So] Source:Protist;167(5):425-439, 2016 Nov.
[Is] ISSN:1618-0941
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:A number of trypanosome isolates from Australian marsupials are within the clade containing the human pathogen Trypanosoma cruzi. Trypanosomes within this clade are thought to have diverged from a common ancestral bat trypanosome. Here, we characterise Trypanosoma noyesi sp. nov. isolated from the critically endangered woylie (Bettongia pencillata) using phylogenetic inferences from three gene regions (18S rDNA, gGAPDH, and CytB) coupled with morphological and behavioural observations in vitro. We also investigated potential vectors and the presence of T. noyesi in the grey-headed flying fox (Pteropus poliocephalus). Phylogenetic analysis revealed T. noyesi and similar genotypes grouped at the periphery of the T. cruzi clade. T. noyesi is morphologically distinct both from other species of Australian trypanosomes and those within the T. cruzi clade. Although trypanosomes were not observed in the digestive tract of ectoparasites and biting flies collected from T. noyesi infected marsupials, tabanid and biting midges tested positive for T. noyesi DNA, indicating they are vector candidates. Tissues from flying foxes were negative for T. noyesi. This study provides novel information on the morphology and genetic variability of an Australian trypanosome within the T. cruzi clade.
[Mh] Termos MeSH primário: Quirópteros
Potoroidae
Trypanosoma/classificação
Tripanossomíase/veterinária
[Mh] Termos MeSH secundário: Animais
Vetores Artrópodes
Dípteros/parasitologia
Proteínas de Protozoários/genética
RNA de Protozoário/genética
RNA Ribossômico 18S/genética
Trypanosoma/genética
Tripanossomíase/epidemiologia
Tripanossomíase/parasitologia
Austrália Ocidental
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Protozoan Proteins); 0 (RNA, Protozoan); 0 (RNA, Ribosomal, 18S)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171108
[Lr] Data última revisão:
171108
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160826
[St] Status:MEDLINE


  3 / 114 MEDLINE  
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[PMID]:27276262
[Au] Autor:Steshenko O; Andrade DM; Honigmann A; Mueller V; Schneider F; Sezgin E; Hell SW; Simons M; Eggeling C
[Ad] Endereço:Cellular Neuroscience, Max Planck Institute of Experimental Medicine, Göttingen, Germany.
[Ti] Título:Reorganization of Lipid Diffusion by Myelin Basic Protein as Revealed by STED Nanoscopy.
[So] Source:Biophys J;110(11):2441-2450, 2016 Jun 07.
[Is] ISSN:1542-0086
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Myelin is a multilayered membrane that ensheathes axonal fibers in the vertebrate nervous system, allowing fast propagation of nerve action potentials. It contains densely packed lipids, lacks an actin-based cytocortex, and requires myelin basic protein (MBP) as its major structural component. This protein is the basic constituent of the proteinaceous meshwork that is localized between adjacent cytoplasmic membranes of the myelin sheath. Yet, it is not clear how MBP influences the organization and dynamics of the lipid constituents of myelin. Here, we used optical stimulated emission depletion super-resolution microscopy in combination with fluorescence correlation spectroscopy to assess the characteristics of diffusion of different fluorescent lipid analogs in myelin membrane sheets of cultured oligodendrocytes and in micrometer-sized domains that were induced by MBP in live epithelial PtK2 cells. Lipid diffusion was significantly faster and less anomalous both in oligodendrocytes and inside the MBP-rich domains of PtK2 cells compared with undisturbed live PtK2 cells. Our data show that MBP reorganizes lipid diffusion, possibly by preventing the buildup of an actin-based cytocortex and by preventing most membrane proteins from entering the myelin sheath region. Yet, in contrast to myelin sheets in oligodendrocytes, the MBP-induced domains in epithelial PtK2 cells demonstrate no change in lipid order, indicating that segregation of long-chain lipids into myelin sheets is a process specific to oligodendrocytes.
[Mh] Termos MeSH primário: Membrana Celular/metabolismo
Etanolaminas/metabolismo
Galactosilceramidas/metabolismo
Proteína Básica da Mielina/metabolismo
Esfingomielinas/metabolismo
[Mh] Termos MeSH secundário: Citoesqueleto de Actina/metabolismo
Animais
Encéfalo/metabolismo
Linhagem Celular
Difusão
Células Epiteliais/metabolismo
Corantes Fluorescentes
Proteínas Luminescentes/genética
Proteínas Luminescentes/metabolismo
Masculino
Camundongos
Microscopia/métodos
Oligodendroglia/metabolismo
Potoroidae
Espectrometria de Fluorescência
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Ethanolamines); 0 (Fluorescent Dyes); 0 (Galactosylceramides); 0 (Luminescent Proteins); 0 (Myelin Basic Protein); 0 (Sphingomyelins); 78A2BX7AEU (phosphorylethanolamine)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170922
[Lr] Data última revisão:
170922
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160609
[St] Status:MEDLINE


  4 / 114 MEDLINE  
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[PMID]:27260808
[Au] Autor:Hing S; Currie A; Broomfield S; Keatley S; Jones K; Thompson RC; Narayan E; Godfrey SS
[Ad] Endereço:Murdoch University, School of Veterinary and Life Sciences, 90 South Street, Murdoch, Western Australia 6150, Australia. Electronic address: S.Hing@murdoch.edu.au.
[Ti] Título:Host stress physiology and Trypanosoma haemoparasite infection influence innate immunity in the woylie (Bettongia penicillata).
[So] Source:Comp Immunol Microbiol Infect Dis;46:32-9, 2016 Jun.
[Is] ISSN:1878-1667
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Understanding immune function is critical to conserving wildlife in view of infectious disease threats, particularly in threatened species vulnerable to stress, immunocompromise and infection. However, few studies examine stress, immune function and infection in wildlife. We used a flow cytometry protocol developed for human infants to assess phagocytosis, a key component of innate immunity, in a critically endangered marsupial, the woylie (Bettongia penicillata). The effects of stress physiology and Trypanosoma infection on phagocytosis were investigated. Blood and faecal samples were collected from woylies in a captive facility over three months. Trypanosoma status was determined using PCR. Faecal cortisol metabolites (FCM) were quantified by enzyme-immunoassay. Mean phagocytosis measured was >90%. An interaction between sex and FCM influenced the percentage of phagocytosing leukocytes, possibly reflecting the influence of sex hormones and glucocorticoids. An interaction between Trypanosoma status and FCM influenced phagocytosis index, suggesting that stress physiology and infection status influence innate immunity.
[Mh] Termos MeSH primário: Imunidade Inata
Fagocitose
Potoroidae/imunologia
Potoroidae/parasitologia
Estresse Fisiológico
Tripanossomíase/veterinária
[Mh] Termos MeSH secundário: Animais
Animais Selvagens/parasitologia
DNA de Protozoário
Fezes/química
Fezes/parasitologia
Interações Hospedeiro-Parasita
Seres Humanos
Hidrocortisona/análise
Filogenia
Potoroidae/fisiologia
RNA Ribossômico 18S
Análise de Sequência de DNA
Fatores Sexuais
Trypanosoma/genética
Trypanosoma/imunologia
Tripanossomíase/imunologia
Tripanossomíase/parasitologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Protozoan); 0 (RNA, Ribosomal, 18S); WI4X0X7BPJ (Hydrocortisone)
[Em] Mês de entrada:1701
[Cu] Atualização por classe:170119
[Lr] Data última revisão:
170119
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160605
[St] Status:MEDLINE


  5 / 114 MEDLINE  
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[PMID]:26864625
[Au] Autor:Fujiwara TK; Iwasawa K; Kalay Z; Tsunoyama TA; Watanabe Y; Umemura YM; Murakoshi H; Suzuki KG; Nemoto YL; Morone N; Kusumi A
[Ad] Endereço:Center for Meso-Bio Single-Molecule Imaging, Institute for Integrated Cell-Material Sciences, Kyoto 606-8501, Japan.
[Ti] Título:Confined diffusion of transmembrane proteins and lipids induced by the same actin meshwork lining the plasma membrane.
[So] Source:Mol Biol Cell;27(7):1101-19, 2016 Apr 01.
[Is] ISSN:1939-4586
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The mechanisms by which the diffusion rate in the plasma membrane (PM) is regulated remain unresolved, despite their importance in spatially regulating the reaction rates in the PM. Proposed models include entrapment in nanoscale noncontiguous domains found in PtK2 cells, slow diffusion due to crowding, and actin-induced compartmentalization. Here, by applying single-particle tracking at high time resolutions, mainly to the PtK2-cell PM, we found confined diffusion plus hop movements (termed "hop diffusion") for both a nonraft phospholipid and a transmembrane protein, transferrin receptor, and equal compartment sizes for these two molecules in all five of the cell lines used here (actual sizes were cell dependent), even after treatment with actin-modulating drugs. The cross-section size and the cytoplasmic domain size both affected the hop frequency. Electron tomography identified the actin-based membrane skeleton (MSK) located within 8.8 nm from the PM cytoplasmic surface of PtK2 cells and demonstrated that the MSK mesh size was the same as the compartment size for PM molecular diffusion. The extracellular matrix and extracellular domains of membrane proteins were not involved in hop diffusion. These results support a model of anchored TM-protein pickets lining actin-based MSK as a major mechanism for regulating diffusion.
[Mh] Termos MeSH primário: Citoesqueleto de Actina
Membrana Celular/metabolismo
Fosfolipídeos/química
Receptores da Transferrina/química
[Mh] Termos MeSH secundário: Animais
Linhagem Celular
Difusão
Seres Humanos
Modelos Biológicos
Potoroidae
Ratos
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Phospholipids); 0 (Receptors, Transferrin)
[Em] Mês de entrada:1701
[Cu] Atualização por classe:170113
[Lr] Data última revisão:
170113
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160212
[St] Status:MEDLINE
[do] DOI:10.1091/mbc.E15-04-0186


  6 / 114 MEDLINE  
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[PMID]:26712388
[Au] Autor:Botero A; Clode PL; Peacock C; Thompson RC
[Ad] Endereço:School of Veterinary and Life Sciences, Murdoch University, South Street, Murdoch, WA 6150, Australia. Electronic address: L.BoteroGomez@murdoch.edu.au.
[Ti] Título:Towards a Better Understanding of the Life Cycle of Trypanosoma copemani.
[So] Source:Protist;167(1):82-92, 2016 Feb.
[Is] ISSN:1618-0941
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:Trypanosoma copemani has been found infecting several threatened/endangered marsupial species within Australia and is thought to be a key player in the rapid decline of the woylie (Bettongia penicillata). To better understand the biology and life cycle of this parasite, the growth requirements, and kinetics of infection of two newly described genotypes, T. copemani G1 and G2, were investigated and compared with the T. cruzi strain-10R26 in vitro. Both G1 and G2 were able to infect all four cell lines tested. The number of infected cells where at least one intracellular amastigote of T. copemani G1 and G2 was seen was below 7% and 15% respectively in most cell lines. However, in VERO cells the rate of infection for T. copemani G2 was 70%-approximately seven and two times higher than for G1 and T. cruzi respectively. Despite the higher infection rate, the number of intracellular forms of T. copemani G2 was lower compared with T. cruzi, and intracellular replicating forms were not observed. The capability of T. copemani G2 to infect cells may have important consequences for pathogenicity and suggests it might employ similar strategies to complete its life cycle in the vertebrate host to those seen in T. cruzi.
[Mh] Termos MeSH primário: Potoroidae
Trypanosoma/fisiologia
Tripanossomíase/veterinária
[Mh] Termos MeSH secundário: Animais
Austrália
Cercopithecus aethiops
Interações Hospedeiro-Parasita
Trypanosoma/genética
Trypanosoma/crescimento & desenvolvimento
Tripanossomíase/parasitologia
Células Vero
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Em] Mês de entrada:1611
[Cu] Atualização por classe:161230
[Lr] Data última revisão:
161230
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:151230
[St] Status:MEDLINE


  7 / 114 MEDLINE  
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[PMID]:26252667
[Au] Autor:Udy DB; Voorhies M; Chan PP; Lowe TM; Dumont S
[Ad] Endereço:Department of Cell & Tissue Biology, University of California San Francisco, San Francisco, CA, United States of America.
[Ti] Título:Draft De Novo Transcriptome of the Rat Kangaroo Potorous tridactylus as a Tool for Cell Biology.
[So] Source:PLoS One;10(8):e0134738, 2015.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The rat kangaroo (long-nosed potoroo, Potorous tridactylus) is a marsupial native to Australia. Cultured rat kangaroo kidney epithelial cells (PtK) are commonly used to study cell biological processes. These mammalian cells are large, adherent, and flat, and contain large and few chromosomes-and are thus ideal for imaging intra-cellular dynamics such as those of mitosis. Despite this, neither the rat kangaroo genome nor transcriptome have been sequenced, creating a challenge for probing the molecular basis of these cellular dynamics. Here, we present the sequencing, assembly and annotation of the draft rat kangaroo de novo transcriptome. We sequenced 679 million reads that mapped to 347,323 Trinity transcripts and 20,079 Unigenes. We present statistics emerging from transcriptome-wide analyses, and analyses suggesting that the transcriptome covers full-length sequences of most genes, many with multiple isoforms. We also validate our findings with a proof-of-concept gene knockdown experiment. We expect that this high quality transcriptome will make rat kangaroo cells a more tractable system for linking molecular-scale function and cellular-scale dynamics.
[Mh] Termos MeSH primário: Biologia Celular
Potoroidae/genética
Transcriptoma/genética
[Mh] Termos MeSH secundário: Animais
Divisão Celular/genética
Linhagem Celular
Disseminação de Informação
Anotação de Sequência Molecular
Isoformas de Proteínas/genética
Isoformas de Proteínas/metabolismo
RNA Mensageiro/genética
RNA Mensageiro/metabolismo
Reprodutibilidade dos Testes
Análise de Sequência de RNA
Especificidade da Espécie
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL
[Nm] Nome de substância:
0 (Protein Isoforms); 0 (RNA, Messenger)
[Em] Mês de entrada:1605
[Cu] Atualização por classe:161019
[Lr] Data última revisão:
161019
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150808
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0134738


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[PMID]:26160545
[Au] Autor:Austen JM; Reid SA; Robinson DR; Friend JA; Ditcham WG; Irwin PJ; Ryan U
[Ad] Endereço:School of Veterinary and Life Sciences,Murdoch University,South Street,Murdoch,Western Australia 6150,Australia.
[Ti] Título:Investigation of the morphological diversity of the potentially zoonotic Trypanosoma copemani in quokkas and Gilbert's potoroos.
[So] Source:Parasitology;142(11):1443-52, 2015 Sep.
[Is] ISSN:1469-8161
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Trypanosomes are blood-borne parasites that can cause severe disease in both humans and animals, yet little is known of the pathogenicity and life-cycles of trypanosomes in native Australian mammals. Trypanosoma copemani is known to be infective to a variety of Australian marsupials and has recently been shown to be potentially zoonotic as it is resistant to normal human serum. In the present study, in vivo and in vitro examination of blood and cultures from Australian marsupials was conducted using light microscopy, immunofluorescence, scanning electron microscopy and fluorescence in situ hybridization. Promastigote, sphaeromastigote and amastigote life-cycle stages were detected in vivo and in vitro. Novel trypanosome-like stages were also detected both in vivo and in vitro representing an oval stage, an extremely thin stage, an adherent stage and a tiny round stage. The tiny round and adherent stages appeared to adhere to erythrocytes causing potential haematological damage with clinical effects similar to haemolytic anaemia. The present study shows for the first time that trypomastigotes are not the only life-cycle stages circulating within the blood stream of trypanosome infected Australian native marsupials and provides insights into possible pathogenic mechanisms of this potentially zoonotic trypanosome species.
[Mh] Termos MeSH primário: Macropodidae/parasitologia
Potoroidae/parasitologia
Trypanosoma/citologia
Tripanossomíase/veterinária
[Mh] Termos MeSH secundário: Animais
Austrália/epidemiologia
Seres Humanos
Estágios do Ciclo de Vida
Trypanosoma/genética
Trypanosoma/crescimento & desenvolvimento
Trypanosoma/isolamento & purificação
Tripanossomíase/epidemiologia
Tripanossomíase/parasitologia
Zoonoses
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Em] Mês de entrada:1609
[Cu] Atualização por classe:151103
[Lr] Data última revisão:
151103
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150711
[St] Status:MEDLINE
[do] DOI:10.1017/S0031182015000785


  9 / 114 MEDLINE  
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[PMID]:26121660
[Au] Autor:Boros G; Miko E; Muramatsu H; Weissman D; Emri E; van der Horst GT; Szegedi A; Horkay I; Emri G; Karikó K; Remenyik É
[Ad] Endereço:Department of Dermatology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
[Ti] Título:Identification of Cyclobutane Pyrimidine Dimer-Responsive Genes Using UVB-Irradiated Human Keratinocytes Transfected with In Vitro-Synthesized Photolyase mRNA.
[So] Source:PLoS One;10(6):e0131141, 2015.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Major biological effects of UVB are attributed to cyclobutane pyrimidine dimers (CPDs), the most common photolesions formed on DNA. To investigate the contribution of CPDs to UVB-induced changes of gene expression, a model system was established by transfecting keratinocytes with pseudouridine-modified mRNA (Ψ-mRNA) encoding CPD-photolyase. Microarray analyses of this model system demonstrated that more than 50% of the gene expression altered by UVB was mediated by CPD photolesions. Functional classification of the gene targets revealed strong effects of CPDs on the regulation of the cell cycle and transcriptional machineries. To confirm the microarray data, cell cycle-regulatory genes, CCNE1 and CDKN2B that were induced exclusively by CPDs were selected for further investigation. Following UVB irradiation, expression of these genes increased significantly at both mRNA and protein levels, but not in cells transfected with CPD-photolyase Ψ-mRNA and exposed to photoreactivating light. Treatment of cells with inhibitors of c-Jun N-terminal kinase (JNK) blocked the UVB-dependent upregulation of both genes suggesting a role for JNK in relaying the signal of UVB-induced CPDs into transcriptional responses. Thus, photolyase mRNA-based experimental platform demonstrates CPD-dependent and -independent events of UVB-induced cellular responses, and, as such, has the potential to identify novel molecular targets for treatment of UVB-mediated skin diseases.
[Mh] Termos MeSH primário: Desoxirribodipirimidina Fotoliase/genética
Regulação da Expressão Gênica/efeitos da radiação
Queratinócitos/metabolismo
Dímeros de Pirimidina/metabolismo
Transfecção
Raios Ultravioleta
[Mh] Termos MeSH secundário: Animais
Linhagem Celular
Ciclina E/genética
Ciclina E/metabolismo
Inibidor de Quinase Dependente de Ciclina p15/genética
Inibidor de Quinase Dependente de Ciclina p15/metabolismo
Reparo do DNA/efeitos da radiação
Desoxirribodipirimidina Fotoliase/metabolismo
Seres Humanos
Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo
Queratinócitos/enzimologia
Queratinócitos/efeitos da radiação
Sistema de Sinalização das MAP Quinases/efeitos da radiação
Análise de Sequência com Séries de Oligonucleotídeos
Proteínas Oncogênicas/genética
Proteínas Oncogênicas/metabolismo
Potoroidae
RNA Mensageiro/genética
RNA Mensageiro/metabolismo
Reação em Cadeia da Polimerase em Tempo Real
Reprodutibilidade dos Testes
Estresse Fisiológico/efeitos da radiação
Transcrição Genética/efeitos da radiação
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (CCNE1 protein, human); 0 (CDKN2B protein, human); 0 (Cyclin E); 0 (Cyclin-Dependent Kinase Inhibitor p15); 0 (Oncogene Proteins); 0 (Pyrimidine Dimers); 0 (RNA, Messenger); EC 2.7.11.24 (JNK Mitogen-Activated Protein Kinases); EC 4.1.99.3 (Deoxyribodipyrimidine Photo-Lyase)
[Em] Mês de entrada:1604
[Cu] Atualização por classe:171121
[Lr] Data última revisão:
171121
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150630
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0131141


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[PMID]:25877871
[Au] Autor:Guan Y; Meurer M; Raghavan S; Rebane A; Lindquist JR; Santos S; Kats I; Davidson MW; Mazitschek R; Hughes TE; Drobizhev M; Knop M; Shah JV
[Ad] Endereço:Department of Systems Biology, Harvard Medical School, Boston, MA 02115 Renal Division, Brigham and Women's Hospital, Boston, MA 02115.
[Ti] Título:Live-cell multiphoton fluorescence correlation spectroscopy with an improved large Stokes shift fluorescent protein.
[So] Source:Mol Biol Cell;26(11):2054-66, 2015 Jun 01.
[Is] ISSN:1939-4586
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:We report an improved variant of mKeima, a monomeric long Stokes shift red fluorescent protein, hmKeima8.5. The increased intracellular brightness and large Stokes shift (∼180 nm) make it an excellent partner with teal fluorescent protein (mTFP1) for multiphoton, multicolor applications. Excitation of this pair by a single multiphoton excitation wavelength (MPE, 850 nm) yields well-separable emission peaks (∼120-nm separation). Using this pair, we measure homo- and hetero-oligomerization interactions in living cells via multiphoton excitation fluorescence correlation spectroscopy (MPE-FCS). Using tandem dimer proteins and small-molecule inducible dimerization domains, we demonstrate robust and quantitative detection of intracellular protein-protein interactions. We also use MPE-FCCS to detect drug-protein interactions in the intracellular environment using a Coumarin 343 (C343)-conjugated drug and hmKeima8.5 as a fluorescence pair. The mTFP1/hmKeima8.5 and C343/hmKeima8.5 combinations, together with our calibration constructs, provide a practical and broadly applicable toolbox for the investigation of molecular interactions in the cytoplasm of living cells.
[Mh] Termos MeSH primário: Corantes Fluorescentes
Proteínas de Fluorescência Verde
Proteínas Luminescentes
Multimerização Proteica
Espectrometria de Fluorescência/métodos
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Animais
Citoplasma
Células HEK293
Seres Humanos
Microscopia de Fluorescência por Excitação Multifotônica
Dados de Sequência Molecular
Potoroidae
Ligação Proteica
Alinhamento de Sequência
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Cyan Fluorescent Protein); 0 (Fluorescent Dyes); 0 (Luminescent Proteins); 0 (red fluorescent protein); 147336-22-9 (Green Fluorescent Proteins)
[Em] Mês de entrada:1603
[Cu] Atualização por classe:161019
[Lr] Data última revisão:
161019
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150417
[St] Status:MEDLINE
[do] DOI:10.1091/mbc.E14-10-1473



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