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[PMID]:29244816
[Au] Autor:Avila-Martin G; Mata-Roig M; Galán-Arriero I; Taylor JS; Busquets X; Escribá PV
[Ad] Endereço:Hospital Nacional de Parapléjicos, Toledo, Spain.
[Ti] Título:Treatment with albumin-hydroxyoleic acid complex restores sensorimotor function in rats with spinal cord injury: Efficacy and gene expression regulation.
[So] Source:PLoS One;12(12):e0189151, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Sensorimotor dysfunction following incomplete spinal cord injury (SCI) is often characterized by paralysis, spasticity and pain. Previously, we showed that intrathecal (i.t.) administration of the albumin-oleic acid (A-OA) complex in rats with SCI produced partial improvement of these symptoms and that oral 2-hydroxyoleic acid (HOA, a non-hydrolyzable OA analogue), was efficacious in the modulation and treatment of nociception and pain-related anxiety, respectively. Here we observed that intrathecal treatment with the complex albumin-HOA (A-HOA) every 3 days following T9 spinal contusion injury improved locomotor function assessed with the Rotarod and inhibited TA noxious reflex activity in Wistar rats. To investigate the mechanism of action of A-HOA, microarray analysis was carried out in the spinal cord lesion area. Representative genes involved in pain and neuroregeneration were selected to validate the changes observed in the microarray analysis by quantitative real-time RT-PCR. Comparison of the expression between healthy rats, SCI rats, and SCI treated with A-HOA rats revealed relevant changes in the expression of genes associated with neuronal morphogenesis and growth, neuronal survival, pain and inflammation. Thus, treatment with A-HOA not only induced a significant overexpression of growth and differentiation factor 10 (GDF10), tenascin C (TNC), aspirin (ASPN) and sushi-repeat-containing X-linked 2 (SRPX2), but also a significant reduction in the expression of prostaglandin E synthase (PTGES) and phospholipases A1 and A2 (PLA1/2). Currently, SCI has very important unmet clinical needs. A-HOA downregulated genes involved with inflammation and upregulated genes involved in neuronal growth, and may serve to promote recovery of function after experimental SCI.
[Mh] Termos MeSH primário: Albuminas/farmacologia
Ácidos Oleicos/farmacologia
Dor/prevenção & controle
Paralisia/tratamento farmacológico
Recuperação de Função Fisiológica/efeitos dos fármacos
Traumatismos da Medula Espinal/tratamento farmacológico
[Mh] Termos MeSH secundário: Albuminas/química
Animais
Esquema de Medicação
Proteínas da Matriz Extracelular/agonistas
Proteínas da Matriz Extracelular/genética
Proteínas da Matriz Extracelular/metabolismo
Regulação da Expressão Gênica
Fator 10 de Diferenciação de Crescimento/agonistas
Fator 10 de Diferenciação de Crescimento/genética
Fator 10 de Diferenciação de Crescimento/metabolismo
Injeções Espinhais
Locomoção/efeitos dos fármacos
Locomoção/fisiologia
Masculino
Proteínas do Tecido Nervoso/agonistas
Proteínas do Tecido Nervoso/genética
Proteínas do Tecido Nervoso/metabolismo
Nociceptividade/efeitos dos fármacos
Ácidos Oleicos/química
Dor/genética
Dor/metabolismo
Dor/patologia
Paralisia/genética
Paralisia/metabolismo
Paralisia/patologia
Fosfolipases/antagonistas & inibidores
Fosfolipases/genética
Fosfolipases/metabolismo
Prostaglandina-E Sintases/antagonistas & inibidores
Prostaglandina-E Sintases/genética
Prostaglandina-E Sintases/metabolismo
Ratos
Ratos Wistar
Recuperação de Função Fisiológica/fisiologia
Teste de Desempenho do Rota-Rod
Medula Espinal/efeitos dos fármacos
Medula Espinal/metabolismo
Medula Espinal/patologia
Traumatismos da Medula Espinal/genética
Traumatismos da Medula Espinal/metabolismo
Traumatismos da Medula Espinal/patologia
Tenascina/agonistas
Tenascina/genética
Tenascina/metabolismo
Resultado do Tratamento
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (2-hydroxyoleic acid); 0 (Albumins); 0 (Extracellular Matrix Proteins); 0 (Gdf10 protein, rat); 0 (Growth Differentiation Factor 10); 0 (Nerve Tissue Proteins); 0 (Oleic Acids); 0 (Tenascin); 0 (asporin protein, rat); EC 3.1.- (Phospholipases); EC 5.3.99.3 (Prostaglandin-E Synthases); EC 5.3.99.3 (Ptges protein, rat)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180108
[Lr] Data última revisão:
180108
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171216
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0189151


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[PMID]:28791824
[Au] Autor:Wojnicz D; Tichaczek-Goska D; Korzekwa K; Kicia M; Hendrich A
[Ad] Endereço:Department of Biology and Medical Parasitology, Wroclaw Medical University, Poland.
[Ti] Título:Anti-enterococcal activities of pentacyclic triterpenes.
[So] Source:Adv Clin Exp Med;26(3):483-490, 2017 May-Jun.
[Is] ISSN:1899-5276
[Cp] País de publicação:Poland
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Asiatic (AA) and ursolic (UA) acids are widely studied phytochemicals, but their antimicrobial properties are still poorly understood. Therefore our research has focused on their activity against uropathogenic Enterococcus faecalis strains. OBJECTIVES: The aim of this research was to determine the influence of AA and UA on the growth, cell morphology, virulence factors and biofilm formation by E. faecalis strains. MATERIAL AND METHODS: AA and UA were purchased from Sigma-Aldrich. E. faecalis strains were isolated from the urine samples of patients with urinary tract infections. The strains were checked for the presence of virulence genes using the PCR method. Their antimicrobial susceptibility was performed using the disc diffusion method. The MICs of triterpenes were determined using the broth microdilution method. The hydrophobicity of cells was established by salt aggregation test. Lipase and lecithinase activities were determined by using an agar medium containing egg yolk emulsion. DNase agar was used for the detection of DNase synthesis. Hemolytic activity was established using a sheep-blood agar. Todd-Hewitt agar medium containing gelatin was used for determination of gelatinase activity. The anti-biofilm activity of asiatic acid and ursolic acid was tested on polystyrene microtiter plates. It was examined using time-kill and biofilm assays. RESULTS: Reduction of growth and enzyme synthesis after exposure of E. faecalis to the acids was observed. None of the acids changed the hydrophobicity of bacteria. Stronger anti-biofilm activity was observed when the bacteria were incubated with AA. Thus, reduction of both the survival and the virulence factors will make bacteria less infectious. CONCLUSIONS: Based on the results obtained, we can assume that the triterpenes investigated should be considered natural components of a human diet rather than as antibacterial agents used on their own.
[Mh] Termos MeSH primário: Antibacterianos/farmacologia
Enterococcus faecalis/efeitos dos fármacos
Triterpenos Pentacíclicos/farmacologia
[Mh] Termos MeSH secundário: Biofilmes/efeitos dos fármacos
Infecções por Bactérias Gram-Positivas/tratamento farmacológico
Infecções por Bactérias Gram-Positivas/metabolismo
Seres Humanos
Lipase/metabolismo
Testes de Sensibilidade Microbiana/métodos
Fosfolipases/metabolismo
Triterpenos/farmacologia
Virulência/efeitos dos fármacos
Fatores de Virulência/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Pentacyclic Triterpenes); 0 (Triterpenes); 0 (Virulence Factors); 9PA5A687X5 (asiatic acid); EC 3.1.- (Phospholipases); EC 3.1.1.3 (Lipase); P3M2575F3F (ursolic acid)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171004
[Lr] Data última revisão:
171004
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170810
[St] Status:MEDLINE
[do] DOI:10.17219/acem/62245


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[PMID]:28758353
[Au] Autor:Yang XY; Li ZQ; Gao ZQ; Wang WJ; Geng Z; Xu JH; She Z; Dong YH
[Ad] Endereço:Key Laboratory of Structural Biology, School of Life Sciences, University of Science and Technology of China, Hefei, China.
[Ti] Título:Structural and SAXS analysis of Tle5-Tli5 complex reveals a novel inhibition mechanism of H2-T6SS in Pseudomonas aeruginosa.
[So] Source:Protein Sci;26(10):2083-2091, 2017 Oct.
[Is] ISSN:1469-896X
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Widely spread in Gram-negative bacteria, the type VI secretion system (T6SS) secretes many effector-immunity protein pairs to help the bacteria compete against other prokaryotic rivals, and infect their eukaryotic hosts. Tle5 and Tle5B are two phospholipase effector protein secreted by T6SS of Pseudomonas aeruginosa. They can facilitate the bacterial internalization process into human epithelial cells by interacting with Akt protein of the PI3K-Akt signal pathway. Tli5 and PA5086-5088 are cognate immunity proteins of Tle5 and Tle5B, respectively. They can interact with their cognate effector proteins to suppress their virulence. Here, we report the crystal structure of Tli5 at 2.8Å resolution and successfully fit it into the Small angle X-ray scattering (SAXS) model of the complete Tle5-Tli5 complex. We identified two important motifs in Tli5 through sequence and structural analysis. One is a conserved loop-ß-hairpin motif that exists in the Tle5 immunity homologs, the other is a long and sharp α-α motif that directly interacts with Tle5 according to SAXS data. We also distinguished the structural features of Tle5 and Tle5B family immunity proteins. Together, our work provided insights into a novel inhibition mechanism that may enhance our understanding of phospholipase D family proteins.
[Mh] Termos MeSH primário: Proteínas de Bactérias/química
Proteínas de Bactérias/metabolismo
Pseudomonas aeruginosa/metabolismo
Sistemas de Secreção Tipo VI/química
Sistemas de Secreção Tipo VI/metabolismo
[Mh] Termos MeSH secundário: Proteínas de Bactérias/genética
Escherichia coli/genética
Modelos Moleculares
Fosfolipases/metabolismo
Pseudomonas aeruginosa/genética
Proteínas Recombinantes/química
Proteínas Recombinantes/genética
Proteínas Recombinantes/metabolismo
Espalhamento a Baixo Ângulo
Eletricidade Estática
Sistemas de Secreção Tipo VI/genética
Difração de Raios X
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (Recombinant Proteins); 0 (Type VI Secretion Systems); EC 3.1.- (Phospholipases)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171010
[Lr] Data última revisão:
171010
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170801
[St] Status:MEDLINE
[do] DOI:10.1002/pro.3246


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[PMID]:28678946
[Au] Autor:Gomes CC; Guimarães LS; Pinto LCC; Camargo GADCG; Valente MIB; Sarquis MIM
[Ad] Endereço:Universidade Federal Fluminense, Faculdade de Odontologia, Departamento de Formação Específica, Nova Friburgo, RJ, Brasil.
[Ti] Título:Investigations of the prevalence and virulence of Candida albicans in periodontal and endodontic lesions in diabetic and normoglycemic patients.
[So] Source:J Appl Oral Sci;25(3):274-281, 2017 May-Jun.
[Is] ISSN:1678-7765
[Cp] País de publicação:Brazil
[La] Idioma:eng
[Ab] Resumo:Objective: The aim of this study was to investigate the prevalence of isolated Candida albicans from periodontal endodontic lesions in diabetic and normoglycemic patients, and the fungi's virulence in different atmospheric conditions. Material and Methods: A case-control study was conducted on 15 patients with type 2 diabetes mellitus (G1) and 15 non-diabetics (G2) with periodontal endodontic lesions. Samples of root canals and periodontal pockets were plated on CHROMagar for later identification by polymerase chain reaction (PCR) and virulence test. Results: C. albicans was identified in 79.2% and 20.8% of the 60 samples collected from diabetic and normoglycemic patients, respectively. Of the 30 samples collected from periodontal pockets, 13 showed a positive culture for C. albicans, with 77% belonging to G1 and 23% to G2. Of the 11 positive samples from root canals, 82% were from G1 and 18% from G2. Production of proteinase presented a precipitation zone Pz<0.63 of 100% in G1 and 72% in G2, in redox and negative (Pz=1), under anaerobic conditions in both groups. Hydrophobicity of the strains from G1 indicated 16.4% with low, 19.3% with moderate, and 64.3% with high hydrophobicity in redox. In G2, 42.2% had low, 39.8% had moderate, 18% had high hydrophobicity in redox. In anaerobic conditions, G1 showed 15.2% with low, 12.8% with moderate, and 72% with high hydrophobicity; in G2, 33.6% had low, 28.8% had moderate, and 37.6% had high hydrophobicity. There was statistical difference in the number of positive cultures between G1 and G2 (p<0.05) with predominance in G1. There was statistical difference for all virulence factors, except hemolysis (p=0.001). Conclusions: Candida albicans was isolated more frequently and had higher virulence in diabetic patients.
[Mh] Termos MeSH primário: Candida albicans/isolamento & purificação
Candida albicans/patogenicidade
Doenças da Polpa Dentária/microbiologia
Diabetes Mellitus Tipo 2/microbiologia
Doenças Periodontais/microbiologia
[Mh] Termos MeSH secundário: Adulto
Idoso
Estudos de Casos e Controles
DNA Fúngico
Cavidade Pulpar/microbiologia
Doenças da Polpa Dentária/diagnóstico por imagem
Doenças da Polpa Dentária/fisiopatologia
Diabetes Mellitus Tipo 2/fisiopatologia
Eletroforese
Feminino
Seres Humanos
Interações Hidrofóbicas e Hidrofílicas
Masculino
Meia-Idade
Oxirredução
Peptídeo Hidrolases/análise
Doenças Periodontais/diagnóstico por imagem
Doenças Periodontais/fisiopatologia
Bolsa Periodontal/microbiologia
Fosfolipases/análise
Reação em Cadeia da Polimerase
Radiografia Dentária
Estatísticas não Paramétricas
Virulência
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Fungal); EC 3.1.- (Phospholipases); EC 3.4.- (Peptide Hydrolases)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170912
[Lr] Data última revisão:
170912
[Sb] Subgrupo de revista:D; IM
[Da] Data de entrada para processamento:170706
[St] Status:MEDLINE


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[PMID]:28559296
[Au] Autor:Anderson MT; Mitchell LA; Mobley HLT
[Ad] Endereço:University of Michigan Medical School, Department of Microbiology and Immunology, Ann Arbor, Michigan, USA.
[Ti] Título:Cysteine Biosynthesis Controls Serratia marcescens Phospholipase Activity.
[So] Source:J Bacteriol;199(16), 2017 Aug 15.
[Is] ISSN:1098-5530
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:causes health care-associated opportunistic infections that can be difficult to treat due to a high incidence of antibiotic resistance. One of the many secreted proteins of is the PhlA phospholipase enzyme. Genes involved in the production and secretion of PhlA were identified by screening a transposon insertion library for phospholipase-deficient mutants on phosphatidylcholine-containing medium. Mutations were identified in four genes ( , , , and ) that are involved in the flagellum-dependent PhlA secretion pathway. An additional phospholipase-deficient isolate harbored a transposon insertion in the gene encoding a predicted serine -acetyltransferase required for cysteine biosynthesis. The requirement for extracellular phospholipase activity was confirmed using a fluorogenic phospholipase substrate. Phospholipase activity was restored to the mutant by the addition of exogenous l-cysteine or -acetylserine to the culture medium and by genetic complementation. Additionally, transcript levels were decreased 6-fold in bacteria lacking and were restored with added cysteine, indicating a role for cysteine-dependent transcriptional regulation of phospholipase activity. mutants also exhibited a defect in swarming motility that was correlated with reduced levels of and flagellar regulator gene transcription. Together, these findings suggest a model in which cysteine is required for the regulation of both extracellular phospholipase activity and surface motility in is known to secrete multiple extracellular enzymes, but PhlA is unusual in that this protein is thought to be exported by the flagellar transport apparatus. In this study, we demonstrate that both extracellular phospholipase activity and flagellar function are dependent on the cysteine biosynthesis pathway. Furthermore, a disruption of cysteine biosynthesis results in decreased and flagellar gene transcription, which can be restored by supplying bacteria with exogenous cysteine. These results identify a previously unrecognized role for CysE and cysteine in the secretion of phospholipase and in bacterial motility.
[Mh] Termos MeSH primário: Cisteína/biossíntese
Fosfolipases/secreção
Serina O-Acetiltransferase/metabolismo
Serratia marcescens/enzimologia
Serratia marcescens/metabolismo
[Mh] Termos MeSH secundário: Meios de Cultura/química
Cisteína/metabolismo
Elementos de DNA Transponíveis
Perfilação da Expressão Gênica
Técnicas de Inativação de Genes
Teste de Complementação Genética
Locomoção
Mutagênese Insercional
Fosfolipases/genética
Serina/análogos & derivados
Serina/metabolismo
Serina O-Acetiltransferase/genética
Serratia marcescens/genética
Serratia marcescens/fisiologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Culture Media); 0 (DNA Transposable Elements); 452VLY9402 (Serine); EC 2.3.1.30 (Serine O-Acetyltransferase); EC 3.1.- (Phospholipases); G05L7T7ZEQ (O-acetylserine); K848JZ4886 (Cysteine)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170810
[Lr] Data última revisão:
170810
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170601
[St] Status:MEDLINE


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[PMID]:28423042
[Au] Autor:Kuo SJ; Yang WH; Liu SC; Tsai CH; Hsu HC; Tang CH
[Ad] Endereço:Graduate Institute of Clinical Medical Science, China Medical University, Taichung, Taiwan.
[Ti] Título:Transforming growth factor ß1 enhances heme oxygenase 1 expression in human synovial fibroblasts by inhibiting microRNA 519b synthesis.
[So] Source:PLoS One;12(4):e0176052, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Osteoarthritis (OA) is manifested by synovial inflammation and cartilage destruction that is directly linked to synovitis, joint swelling and pain. In the light of the role of synovium in the pathogenesis and the symptoms of OA, synovium-targeted therapy is a promising strategy to mitigate the symptoms and progression of OA. Transforming growth factor beta 1 (TGF-ß1), a secreted homodimeric protein, possesses unique and potent anti-inflammatory and immune-regulatory properties in many cell types. Heme oxygenase 1 (HO-1) is an inducible anti-inflammatory and stress responsive enzyme that has been proven to prevent injuries caused by many diseases. Despite the similar anti-inflammatory profile and their involvement in the pathogenesis of arthritic diseases, no studies have as yet explored the possibility of any association between the expression of TGF-ß1 and HO-1. METHODOLOGY/PRINCIPAL FINDINGS: TGF-ß1-induced HO-1 expression was examined by HO-1 promoter assay, qPCR, and Western blotting. The siRNAs and enzyme inhibitors were utilized to determine the intermediate involved in the signal transduction pathway. We showed that TGF-ß1 stimulated the synthesis of HO-1 in a concentration- and time-dependent manner, which can be mitigated by blockade of the phospholipase (PLC)γ/protein kinase C alpha (PKC)α pathway. We also showed that the expression of miRNA-519b, which blocks HO-1 transcription, is inhibited by TGF-ß1, and the suppression of miRNA 519b could be reversed via blockade of the PLCγ/PKCα pathway. CONCLUSIONS/SIGNIFICANCE: TGF-ß1 stimulated the expression of HO-1 via activating the PLCγ/PKCα pathway and suppressing the downstream expression of miRNA-519b. These results may shed light on the pathogenesis and treatment of OA.
[Mh] Termos MeSH primário: Fibroblastos/metabolismo
Heme Oxigenase-1/genética
MicroRNAs/genética
Osteoartrite/genética
Fator de Crescimento Transformador beta1/genética
[Mh] Termos MeSH secundário: Anticorpos/farmacologia
Artroplastia do Joelho
Carbazóis/farmacologia
Cartilagem Articular/efeitos dos fármacos
Cartilagem Articular/metabolismo
Cartilagem Articular/patologia
Inibidores Enzimáticos/farmacologia
Fibroblastos/efeitos dos fármacos
Fibroblastos/patologia
Regulação da Expressão Gênica
Heme Oxigenase-1/metabolismo
Seres Humanos
Indóis/farmacologia
Maleimidas/farmacologia
MicroRNAs/metabolismo
Osteoartrite/metabolismo
Osteoartrite/patologia
Osteoartrite/cirurgia
Fosfolipases/antagonistas & inibidores
Fosfolipases/genética
Fosfolipases/metabolismo
Cultura Primária de Células
Proteína Quinase C-alfa/antagonistas & inibidores
Proteína Quinase C-alfa/genética
Proteína Quinase C-alfa/metabolismo
RNA Interferente Pequeno/genética
RNA Interferente Pequeno/metabolismo
Proteínas Recombinantes/genética
Proteínas Recombinantes/metabolismo
Proteínas Recombinantes/farmacologia
Transdução de Sinais
Membrana Sinovial/efeitos dos fármacos
Membrana Sinovial/metabolismo
Membrana Sinovial/patologia
Fator de Crescimento Transformador beta1/metabolismo
Fator de Crescimento Transformador beta1/farmacologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies); 0 (Carbazoles); 0 (Enzyme Inhibitors); 0 (Indoles); 0 (MIRN519 microRNA, human); 0 (Maleimides); 0 (MicroRNAs); 0 (RNA, Small Interfering); 0 (Recombinant Proteins); 0 (TGFB1 protein, human); 0 (Transforming Growth Factor beta1); 136194-77-9 (Go 6976); EC 1.14.14.18 (HMOX1 protein, human); EC 1.14.14.18 (Heme Oxygenase-1); EC 2.7.11.13 (Protein Kinase C-alpha); EC 3.1.- (Phospholipases); L79H6N0V6C (bisindolylmaleimide I)
[Em] Mês de entrada:1704
[Cu] Atualização por classe:170505
[Lr] Data última revisão:
170505
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170420
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0176052


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[PMID]:28419631
[Au] Autor:Lévêque MF; Berry L; Yamaryo-Botté Y; Nguyen HM; Galera M; Botté CY; Besteiro S
[Ad] Endereço:DIMNP - UMR5235, CNRS, Université de Montpellier, Montpellier, France.
[Ti] Título:TgPL2, a patatin-like phospholipase domain-containing protein, is involved in the maintenance of apicoplast lipids homeostasis in Toxoplasma.
[So] Source:Mol Microbiol;105(1):158-174, 2017 07.
[Is] ISSN:1365-2958
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Patatin-like phospholipases are involved in numerous cellular functions, including lipid metabolism and membranes remodeling. The patatin-like catalytic domain, whose phospholipase activity relies on a serine-aspartate dyad and an anion binding box, is widely spread among prokaryotes and eukaryotes. We describe TgPL2, a novel patatin-like phospholipase domain-containing protein from the parasitic protist Toxoplasma gondii. TgPL2 is a large protein, in which the key motifs for enzymatic activity are conserved in the patatin-like domain. Using immunofluorescence assays and immunoelectron microscopy analysis, we have shown that TgPL2 localizes to the apicoplast, a non-photosynthetic plastid found in most apicomplexan parasites. This plastid hosts several important biosynthetic pathways, which makes it an attractive organelle for identifying new potential drug targets. We thus addressed TgPL2 function by generating a conditional knockdown mutant and demonstrated it has an essential contribution for maintaining the integrity of the plastid. In absence of TgPL2, the organelle is rapidly lost and remaining apicoplasts appear enlarged, with an abnormal accumulation of membranous structures, suggesting a defect in lipids homeostasis. More precisely, analyses of lipid content upon TgPL2 depletion suggest this protein is important for maintaining levels of apicoplast-generated fatty acids, and also regulating phosphatidylcholine and lysophosphatidylcholine levels in the parasite.
[Mh] Termos MeSH primário: Apicoplastos/metabolismo
Fosfolipases/metabolismo
Toxoplasma/metabolismo
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Animais
Apicoplastos/genética
Sequência de Bases
Domínio Catalítico
Ácidos Graxos/metabolismo
Homeostase
Metabolismo dos Lipídeos/fisiologia
Lipídeos
Parasitos
Plastídeos/metabolismo
Domínios Proteicos
Proteínas de Protozoários/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Fatty Acids); 0 (Lipids); 0 (Protozoan Proteins); EC 3.1.- (Phospholipases)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171031
[Lr] Data última revisão:
171031
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170419
[St] Status:MEDLINE
[do] DOI:10.1111/mmi.13694


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[PMID]:28280998
[Au] Autor:Alves F; de Oliveira Mima EG; Passador RCP; Bagnato VS; Jorge JH; Pavarina AC
[Ad] Endereço:Department of Dental Materials and Prosthodontics, Araraquara Dental School, UNESP - Univ. Estadual Paulista, Rua Humaitá, 1680, 14801-903, Araraquara, SP, Brazil.
[Ti] Título:Virulence factors of fluconazole-susceptible and fluconazole-resistant Candida albicans after antimicrobial photodynamic therapy.
[So] Source:Lasers Med Sci;32(4):815-826, 2017 May.
[Is] ISSN:1435-604X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:This study evaluated the effects of antimicrobial photodynamic therapy (aPDT) mediated by Photodithazine® (PDZ) and LED light on the virulence factors of fluconazole-susceptible (CaS) and fluconazole-resistant (CaR) Candida albicans. Standardized suspensions of strains were prepared (10 ), and after 48 h of biofilm formation, these strains were incubated with PDZ (100 mg/L) for 20 min and exposed to LED light (660 nm, 37.5 J/cm ). Additional samples were treated with PDZ or light only, and the control consisted of biofilms that received no treatment. After aPDT, the cells were recovered and the virulence factors were evaluated. To analyze the capacity of adhesion, cells were recovered after aPDT and submitted to the adhesion process in the bottom of a 96-well plate. After this, metabolic activity tests (XTT assay) and cell viability (colony forming units per milliliter, CFU/mL) were applied. To evaluate the biofilm-forming ability after aPDT, the cells recovered were submitted to biofilm formation procedures, and the biofilm formed was evaluated by XTT, CFU/mL, and total biomass (crystal violet) tests. Lastly, the capacity for synthesizing protease and phospholipase enzymes after aPDT was evaluated by fluorimetric tests. Data were analyzed by two- or three-way ANOVA tests (p ≤ 0.05). It was verified that aPDT reduced the viability of both strains, fluconazole-susceptible and fluconazole-resistant C. albicans. It was also observed that the CaR strain had lower susceptibility to the aPDT when compared with the CaS strain. However, regarding the virulence factors evaluated, it was demonstrated that aPDT did not alter the adherence and biofilm formation ability and enzymatic production.
[Mh] Termos MeSH primário: Anti-Infecciosos/farmacologia
Antifúngicos/farmacologia
Candida albicans/efeitos dos fármacos
Candida albicans/patogenicidade
Farmacorresistência Fúngica/efeitos dos fármacos
Fluconazol/farmacologia
Fotoquimioterapia/métodos
Fatores de Virulência/metabolismo
[Mh] Termos MeSH secundário: Adesividade
Biofilmes/efeitos dos fármacos
Biomassa
Testes de Sensibilidade Microbiana
Peptídeo Hidrolases/metabolismo
Fosfolipases/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Infective Agents); 0 (Antifungal Agents); 0 (Virulence Factors); 8VZV102JFY (Fluconazole); EC 3.1.- (Phospholipases); EC 3.4.- (Peptide Hydrolases)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:171013
[Lr] Data última revisão:
171013
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170311
[St] Status:MEDLINE
[do] DOI:10.1007/s10103-017-2177-y


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[PMID]:28228439
[Au] Autor:Gilles LM; Khaled A; Laffaire JB; Chaignon S; Gendrot G; Laplaige J; Bergès H; Beydon G; Bayle V; Barret P; Comadran J; Martinant JP; Rogowsky PM; Widiez T
[Ad] Endereço:Laboratoire Reproduction et Développement des Plantes, Univ Lyon ENS de Lyon UCB Lyon 1 CNRS, INRA, Lyon, France.
[Ti] Título:Loss of pollen-specific phospholipase NOT LIKE DAD triggers gynogenesis in maize.
[So] Source:EMBO J;36(6):707-717, 2017 Mar 15.
[Is] ISSN:1460-2075
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Gynogenesis is an asexual mode of reproduction common to animals and plants, in which stimuli from the sperm cell trigger the development of the unfertilized egg cell into a haploid embryo. Fine mapping restricted a major maize QTL (quantitative trait locus) responsible for the aptitude of inducer lines to trigger gynogenesis to a zone containing a single gene ( ) coding for a patatin-like phospholipase A. In all surveyed inducer lines, carries a 4-bp insertion leading to a predicted truncated protein. This frameshift mutation is responsible for haploid induction because complementation with wild-type abolishes the haploid induction capacity. Activity of the promoter is restricted to mature pollen and pollen tube. The translational NLD::citrine fusion protein likely localizes to the sperm cell plasma membrane. In roots, the truncated protein is no longer localized to the plasma membrane, contrary to the wild-type NLD protein. In conclusion, an intact pollen-specific phospholipase is required for successful sexual reproduction and its targeted disruption may allow establishing powerful haploid breeding tools in numerous crops.
[Mh] Termos MeSH primário: Óvulo Vegetal/crescimento & desenvolvimento
Fosfolipases/metabolismo
Proteínas de Plantas/metabolismo
Pólen/enzimologia
Reprodução
Zea mays/fisiologia
[Mh] Termos MeSH secundário: Regulação da Expressão Gênica de Plantas
Fosfolipases/deficiência
Zea mays/enzimologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Plant Proteins); EC 3.1.- (Phospholipases)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170713
[Lr] Data última revisão:
170713
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170224
[St] Status:MEDLINE
[do] DOI:10.15252/embj.201796603


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[PMID]:28114299
[Au] Autor:Kelliher T; Starr D; Richbourg L; Chintamanani S; Delzer B; Nuccio ML; Green J; Chen Z; McCuiston J; Wang W; Liebler T; Bullock P; Martin B
[Ad] Endereço:Seeds Research, Syngenta Crop Protection, 9 Davis Drive, Research Triangle Park, North Carolina 27709, USA.
[Ti] Título:MATRILINEAL, a sperm-specific phospholipase, triggers maize haploid induction.
[So] Source:Nature;542(7639):105-109, 2017 02 02.
[Is] ISSN:1476-4687
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Sexual reproduction in flowering plants involves double fertilization, the union of two sperm from pollen with two sex cells in the female embryo sac. Modern plant breeders increasingly seek to circumvent this process to produce doubled haploid individuals, which derive from the chromosome-doubled cells of the haploid gametophyte. Doubled haploid production fixes recombinant haploid genomes in inbred lines, shaving years off the breeding process. Costly, genotype-dependent tissue culture methods are used in many crops, while seed-based in vivo doubled haploid systems are rare in nature and difficult to manage in breeding programmes. The multi-billion-dollar maize hybrid seed business, however, is supported by industrial doubled haploid pipelines using intraspecific crosses to in vivo haploid inducer males derived from Stock 6, first reported in 1959 (ref. 5), followed by colchicine treatment. Despite decades of use, the mode of action remains controversial. Here we establish, through fine mapping, genome sequencing, genetic complementation, and gene editing, that haploid induction in maize (Zea mays) is triggered by a frame-shift mutation in MATRILINEAL (MTL), a pollen-specific phospholipase, and that novel edits in MTL lead to a 6.7% haploid induction rate (the percentage of haploid progeny versus total progeny). Wild-type MTL protein localizes exclusively to sperm cytoplasm, and pollen RNA-sequence profiling identifies a suite of pollen-specific genes overexpressed during haploid induction, some of which may mediate the formation of haploid seed. These findings highlight the importance of male gamete cytoplasmic components to reproductive success and male genome transmittance. Given the conservation of MTL in the cereals, this discovery may enable development of in vivo haploid induction systems to accelerate breeding in crop plants.
[Mh] Termos MeSH primário: Mutação da Fase de Leitura
Haploidia
Fosfolipases/genética
Fosfolipases/metabolismo
Pólen/enzimologia
Zea mays/enzimologia
Zea mays/genética
[Mh] Termos MeSH secundário: Alelos
Cruzamento/métodos
Citoplasma/enzimologia
Fertilização
Edição de Genes
Regulação da Expressão Gênica de Plantas
Genes de Plantas/genética
Teste de Complementação Genética
Fenótipo
Proteínas de Plantas/metabolismo
Pólen/citologia
Pólen/genética
Sementes/genética
Análise de Sequência de RNA
Zea mays/citologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Plant Proteins); EC 3.1.- (Phospholipases)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170713
[Lr] Data última revisão:
170713
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170124
[St] Status:MEDLINE
[do] DOI:10.1038/nature20827



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