Base de dados : MEDLINE
Pesquisa : B01.268 [Categoria DeCS]
Referências encontradas : 39 [refinar]
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[PMID]:29256280
[Au] Autor:Szoke K; Sándor AD; Boldogh SA; Görföl T; Votýpka J; Takács N; Estók P; Kováts D; Corduneanu A; Molnár V; Kontschán J; Hornok S
[Ad] Endereço:1 Department of Parasitology and Zoology, University of Veterinary Medicine , István u. 2, H-1078 Budapest , Hungary.
[Ti] Título:DNA of free-living bodonids (Euglenozoa: Kinetoplastea) in bat ectoparasites: potential relevance to the evolution of parasitic trypanosomatids.
[So] Source:Acta Vet Hung;65(4):531-540, 2017 12.
[Is] ISSN:0236-6290
[Cp] País de publicação:Hungary
[La] Idioma:eng
[Ab] Resumo:Kinetoplastids are flagellated protozoa, including principally free-living bodonids and exclusively parasitic trypanosomatids. In the most species-rich genus, Trypanosoma, more than thirty species were found to infect bats worldwide. Bat trypanosomes are also known to have played a significant role in the evolution of T. cruzi, a species with high veterinary medical significance. Although preliminary data attested the occurrence of bat trypanosomes in Hungary, these were never sought for with molecular methods. Therefore, amplification of an approx. 900-bp fragment of the 18S rRNA gene of kinetoplastids was attempted from 307 ixodid and 299 argasid ticks collected from bats, and from 207 cimicid bugs collected from or near bats in Hungary and Romania. Three samples, one per each bat ectoparasite group, were PCR positive. Sequencing revealed the presence of DNA from free-living bodonids (Bodo saltans and neobodonids), but no trypanosomes were detected. The most likely source of bodonid DNA detected here in engorged bat ectoparasites is the blood of their bat hosts. However, how bodonids were acquired by bats, can only be speculated. Bats are known to drink from freshwater bodies, i.e. the natural habitats of B. saltans and related species, allowing bats to ingest bodonids. Consequently, these results suggest that at least the DNA of bodonids might pass through the alimentary mucosa of bats into their circulation. The above findings highlight the importance of studying bats and other mammals for the occurrence of bodonids in their blood and excreta, with potential relevance to the evolution of free-living kinetoplastids towards parasitism.
[Mh] Termos MeSH primário: Evolução Biológica
Quirópteros/parasitologia
DNA/genética
Ectoparasitoses/veterinária
Euglenozoários/genética
Trypanosomatina/genética
[Mh] Termos MeSH secundário: Animais
Cimicidae/parasitologia
Ectoparasitoses/parasitologia
Filogeografia
Carrapatos/parasitologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
9007-49-2 (DNA)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180305
[Lr] Data última revisão:
180305
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171220
[St] Status:MEDLINE
[do] DOI:10.1556/004.2017.051


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[PMID]:28386020
[Au] Autor:Venkatesh D; Boehm C; Barlow LD; Nankissoor NN; O'Reilly A; Kelly S; Dacks JB; Field MC
[Ad] Endereço:Wellcome Trust Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, UK.
[Ti] Título:Evolution of the endomembrane systems of trypanosomatids - conservation and specialisation.
[So] Source:J Cell Sci;130(8):1421-1434, 2017 Apr 15.
[Is] ISSN:1477-9137
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Parasite surfaces support multiple functions required for survival within their hosts, and maintenance and functionality of the surface depends on membrane trafficking. To understand the evolutionary history of trypanosomatid trafficking, where multiple lifestyles and mechanisms of host interactions are known, we examined protein families central to defining intracellular compartments and mediating transport, namely Rabs, SNAREs and RabGAPs, across all available Euglenozoa genomes. Bodonids possess a large trafficking repertoire, which is mainly retained by the group, with extensive losses in other lineages, particularly African trypanosomes and phytomonads. There are no large-scale expansions or contractions from an inferred ancestor, excluding direct associations between parasitism or host range. However, we observe stepwise secondary losses within Rab and SNARE cohorts (but not RabGAPs). Major changes are associated with endosomal and late exocytic pathways, consistent with the diversity in surface proteomes between trypanosomatids and mechanisms of interaction with the host. Along with the conserved core family proteins, several lineage-specific members of the Rab (but not SNARE) family were found. Significantly, testing predictions of SNARE complex composition by proteomics confirms generalised retention of function across eukaryotes.
[Mh] Termos MeSH primário: Evolução Biológica
Membrana Celular/metabolismo
Euglenozoários
Interações Hospedeiro-Patógeno
Proteínas de Protozoários/metabolismo
Trypanosoma cruzi
Proteínas rab de Ligação ao GTP/metabolismo
[Mh] Termos MeSH secundário: Sequência Conservada/genética
Endocitose
Exocitose
Proteínas Ativadoras de GTPase/genética
Proteínas Ativadoras de GTPase/metabolismo
Genoma
Especificidade de Hospedeiro
Transporte Proteico
Proteômica
Proteínas de Protozoários/genética
Proteínas SNARE/genética
Proteínas SNARE/metabolismo
Especificidade da Espécie
Proteínas rab de Ligação ao GTP/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (GTPase-Activating Proteins); 0 (Protozoan Proteins); 0 (SNARE Proteins); EC 3.6.5.2 (rab GTP-Binding Proteins)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:170518
[Lr] Data última revisão:
170518
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170408
[St] Status:MEDLINE
[do] DOI:10.1242/jcs.197640


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[PMID]:27925020
[Au] Autor:Oliveira SS; Gonçalves DS; Garcia-Gomes AD; Gonçalves IC; Seabra SH; Menna-Barreto RF; Lopes AH; D'Avila-Levy CM; Santos AL; Branquinha MH
[Ad] Endereço:Universidade Federal do Rio de Janeiro, Instituto de Microbiologia Paulo de Góes, Departamento de Microbiologia Geral, Laboratório de Investigação de Peptidases, Rio de Janeiro, RJ, Brasil.
[Ti] Título:Susceptibility of Phytomonas serpens to calpain inhibitors in vitro: interference on the proliferation, ultrastructure, cysteine peptidase expression and interaction with the invertebrate host.
[So] Source:Mem Inst Oswaldo Cruz;112(1):31-43, 2017 Jan 01.
[Is] ISSN:1678-8060
[Cp] País de publicação:Brazil
[La] Idioma:eng
[Ab] Resumo:A pleiotropic response to the calpain inhibitor MDL28170 was detected in the tomato parasite Phytomonas serpens. Ultrastructural studies revealed that MDL28170 caused mitochondrial swelling, shortening of flagellum and disruption of trans Golgi network. This effect was correlated to the inhibition in processing of cruzipain-like molecules, which presented an increase in expression paralleled by decreased proteolytic activity. Concomitantly, a calcium-dependent cysteine peptidase was detected in the parasite extract, the activity of which was repressed by pre-incubation of parasites with MDL28170. Flow cytometry and Western blotting analyses revealed the differential expression of calpain-like proteins (CALPs) in response to the pre-incubation of parasites with the MDL28170, and confocal fluorescence microscopy confirmed their surface location. The interaction of promastigotes with explanted salivary glands of the insect Oncopeltus fasciatus was reduced when parasites were pre-treated with MDL28170, which was correlated to reduced levels of surface cruzipain-like and gp63-like molecules. Treatment of parasites with anti-Drosophila melanogaster (Dm) calpain antibody also decreased the adhesion process. Additionally, parasites recovered from the interaction process presented higher levels of surface cruzipain-like and gp63-like molecules, with similar levels of CALPs cross-reactive to anti-Dm-calpain antibody. The results confirm the importance of exploring the use of calpain inhibitors in studying parasites' physiology.
[Mh] Termos MeSH primário: Cisteína/efeitos dos fármacos
Euglenozoários/efeitos dos fármacos
Heterópteros/parasitologia
Interações Hospedeiro-Parasita/fisiologia
[Mh] Termos MeSH secundário: Animais
Western Blotting
Cisteína/metabolismo
Dipeptídeos
Euglenozoários/enzimologia
Euglenozoários/ultraestrutura
Citometria de Fluxo
Dose Letal Mediana
Microscopia Eletrônica
Glândulas Salivares/parasitologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Dipeptides); 88191-84-8 (calpain inhibitor III); K848JZ4886 (Cysteine)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170731
[Lr] Data última revisão:
170731
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161208
[St] Status:MEDLINE


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[PMID]:27889663
[Au] Autor:Cavalier-Smith T
[Ad] Endereço:Department of Zoology, University of Oxford, South Parks Road, Oxford OX1 3PS, UK. Electronic address: tom.cavalier-smith@zoo.ox.ac.uk.
[Ti] Título:Higher classification and phylogeny of Euglenozoa.
[So] Source:Eur J Protistol;56:250-276, 2016 Oct.
[Is] ISSN:1618-0429
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:Discoveries of numerous new taxa and advances in ultrastructure and sequence phylogeny (including here the first site-heterogeneous 18S rDNA trees) require major improvements to euglenozoan higher-level taxonomy. I therefore divide Euglenozoa into three subphyla of substantially different body plans: Euglenoida with pellicular strips; anaerobic Postgaardia (class Postgaardea) dependent on surface bacteria and with uniquely modified feeding apparatuses; and new subphylum Glycomonada characterised by glycosomes (Kinetoplastea, Diplonemea). Euglenoida comprise two new infraphyla: Entosiphona with three feeding rods and Dipilida ancestrally with two. Dipilida comprise basal superclass Rigimonada with longitudinal rigid strips [i.e. new classes Stavomonadea (Petalomonadida, Decastavida and new order Heterostavida) and Ploeotarea (Ploeotiida) with contrasting oral cytoskeletons] and derived superclass Spirocuta with more numerous spirally arranged, often slideable, strips (clade Peranemea/Euglenophyceae) and a different, highly conserved microtubule pattern at strip joints. Peranemea comprise four orders: Peranemida (anterior gliding, protrusible rods), and three new, Anisonemida (posterior gliders), Natomonadida (swimmers including phagotrophic new suborder Metanemina and osmotrophic suborder Rhabdomonadina), and Acroglissida (anterior gliders with cytoproct). I establish orders Entosiphonida, Rapazida, Bihospitida; and seven new euglenoid families (Entosiphonidae, peranemean Neometanemidae, Rapazidae, two stavomonad, two ploeotiid) and three new postgaardian, and three kinetoplastid families (Neobodonidae, Rhynchomonadidae, Parabodonidae), plus new diplonemid family Hemistasiidae for Hemistasia.
[Mh] Termos MeSH primário: Euglenozoários/classificação
Filogenia
[Mh] Termos MeSH secundário: Euglenozoários/citologia
Euglenozoários/genética
RNA Ribossômico 18S/genética
Especificidade da Espécie
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Ribosomal, 18S)
[Em] Mês de entrada:1612
[Cu] Atualização por classe:161230
[Lr] Data última revisão:
161230
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161128
[St] Status:MEDLINE


  5 / 39 MEDLINE  
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[PMID]:27715490
[Au] Autor:Valach M; Moreira S; Faktorová D; Lukes J; Burger G
[Ad] Endereço:a Department of Biochemistry and Robert-Cedergren , Center for Bioinformatics and Genomics, Université de Montréal , Montreal , Canada.
[Ti] Título:Post-transcriptional mending of gene sequences: Looking under the hood of mitochondrial gene expression in diplonemids.
[So] Source:RNA Biol;13(12):1204-1211, 2016 Dec.
[Is] ISSN:1555-8584
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The instructions to make proteins and structural RNAs are laid down in gene sequences. Yet, in certain instances, these primary instructions need to be modified considerably during gene expression, most often at the transcript level. Here we review a case of massive post-transcriptional revisions via trans-splicing and RNA editing, a phenomenon occurring in mitochondria of a recently recognized protist group, the diplonemids. As of now, the various post-transcriptional steps have been cataloged in detail, but how these processes function is still unknown. Since genetic manipulation techniques such as gene replacement and RNA interference have not yet been established for these organisms, alternative strategies have to be deployed. Here, we discuss the experimental and bioinformatics approaches that promise to unravel the molecular machineries of trans-splicing and RNA editing in Diplonema mitochondria.
[Mh] Termos MeSH primário: Euglenozoários/genética
Mitocôndrias/genética
Proteínas Mitocondriais/genética
RNA de Transferência/metabolismo
[Mh] Termos MeSH secundário: Sequência de Bases
Regulação da Expressão Gênica
Proteínas de Protozoários/genética
Edição de RNA
Processamento Pós-Transcricional do RNA
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Nm] Nome de substância:
0 (Mitochondrial Proteins); 0 (Protozoan Proteins); 9014-25-9 (RNA, Transfer)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171018
[Lr] Data última revisão:
171018
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161008
[St] Status:MEDLINE


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[PMID]:27170716
[Au] Autor:Morales J; Hashimoto M; Williams TA; Hirawake-Mogi H; Makiuchi T; Tsubouchi A; Kaga N; Taka H; Fujimura T; Koike M; Mita T; Bringaud F; Concepción JL; Hashimoto T; Embley TM; Nara T
[Ad] Endereço:Department of Molecular and Cellular Parasitology, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan.
[Ti] Título:Differential remodelling of peroxisome function underpins the environmental and metabolic adaptability of diplonemids and kinetoplastids.
[So] Source:Proc Biol Sci;283(1830), 2016 05 11.
[Is] ISSN:1471-2954
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The remodelling of organelle function is increasingly appreciated as a central driver of eukaryotic biodiversity and evolution. Kinetoplastids including Trypanosoma and Leishmania have evolved specialized peroxisomes, called glycosomes. Glycosomes uniquely contain a glycolytic pathway as well as other enzymes, which underpin the physiological flexibility of these major human pathogens. The sister group of kinetoplastids are the diplonemids, which are among the most abundant eukaryotes in marine plankton. Here we demonstrate the compartmentalization of gluconeogenesis, or glycolysis in reverse, in the peroxisomes of the free-living marine diplonemid, Diplonema papillatum Our results suggest that peroxisome modification was already under way in the common ancestor of kinetoplastids and diplonemids, and raise the possibility that the central importance of gluconeogenesis to carbon metabolism in the heterotrophic free-living ancestor may have been an important selective driver. Our data indicate that peroxisome modification is not confined to the kinetoplastid lineage, but has also been a factor in the success of their free-living euglenozoan relatives.
[Mh] Termos MeSH primário: Euglenozoários/citologia
Euglenozoários/metabolismo
Peroxissomos/metabolismo
Trypanosoma cruzi/citologia
[Mh] Termos MeSH secundário: Aminoácidos/metabolismo
Carbono/metabolismo
Enzimas/metabolismo
Euglenozoários/genética
Gluconeogênese
Microcorpos
Via de Pentose Fosfato
Filogenia
Transdução de Sinais
Trypanosoma cruzi/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Amino Acids); 0 (Enzymes); 7440-44-0 (Carbon)
[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:160513
[St] Status:MEDLINE


  7 / 39 MEDLINE  
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[PMID]:27001515
[Au] Autor:Moreira S; Valach M; Aoulad-Aissa M; Otto C; Burger G
[Ad] Endereço:Department of Biochemistry and Robert-Cedergren Centre for Bioinformatics and Genomics; Université de Montréal, Montreal, H3C 3J7, Canada.
[Ti] Título:Novel modes of RNA editing in mitochondria.
[So] Source:Nucleic Acids Res;44(10):4907-19, 2016 Jun 02.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Gene structure and expression in diplonemid mitochondria are unparalleled. Genes are fragmented in pieces (modules) that are separately transcribed, followed by the joining of module transcripts to contiguous RNAs. Some instances of unique uridine insertion RNA editing at module boundaries were noted, but the extent and potential occurrence of other editing types remained unknown. Comparative analysis of deep transcriptome and genome data from Diplonema papillatum mitochondria reveals ∼220 post-transcriptional insertions of uridines, but no insertions of other nucleotides nor deletions. In addition, we detect in total 114 substitutions of cytosine by uridine and adenosine by inosine, amassed into unusually compact clusters. Inosines in transcripts were confirmed experimentally. This is the first report of adenosine-to-inosine editing of mRNAs and ribosomal RNAs in mitochondria. In mRNAs, editing causes mostly amino-acid additions and non-synonymous substitutions; in ribosomal RNAs, it permits formation of canonical secondary structures. Two extensively edited transcripts were compared across four diplonemids. The pattern of uridine-insertion editing is strictly conserved, whereas substitution editing has diverged dramatically, but still rendering diplonemid proteins more similar to other eukaryotic orthologs. We posit that RNA editing not only compensates but also sustains, or even accelerates, ultra-rapid evolution of genome structure and sequence in diplonemid mitochondria.
[Mh] Termos MeSH primário: Euglenozoários/genética
Mitocôndrias/genética
Edição de RNA
RNA/metabolismo
[Mh] Termos MeSH secundário: Adenosina/metabolismo
Desaminação
Euglenozoários/metabolismo
Genes Mitocondriais
Genes de RNAr
Inosina/metabolismo
RNA/química
Trans-Splicing
Transcriptoma
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, mitochondrial); 5A614L51CT (Inosine); 63231-63-0 (RNA); K72T3FS567 (Adenosine)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:170526
[Lr] Data última revisão:
170526
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160323
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkw188


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[PMID]:26449279
[Au] Autor:Moreira S; Noutahi E; Lamoureux G; Burger G
[Ad] Endereço:Department of Biochemistry and Robert-Cedergren Centre for Bioinformatics and Genomics, Université de Montréal, Montreal, QC, Canada. sandrine.moreira@umontreal.ca.
[Ti] Título:Three-dimensional structure model and predicted ATP interaction rewiring of a deviant RNA ligase 2.
[So] Source:BMC Struct Biol;15:20, 2015 Oct 09.
[Is] ISSN:1472-6807
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: RNA ligases 2 are scarce and scattered across the tree of life. Two members of this family are well studied: the mitochondrial RNA editing ligase from the parasitic trypanosomes (Kinetoplastea), a promising drug target, and bacteriophage T4 RNA ligase 2, a workhorse in molecular biology. Here we report the identification of a divergent RNA ligase 2 (DpRNL) from Diplonema papillatum (Diplonemea), a member of the kinetoplastids' sister group. METHODS: We identified DpRNL with methods based on sensitive hidden Markov Model. Then, using homology modeling and molecular dynamics simulations, we established a three dimensional structure model of DpRNL complexed with ATP and Mg2+. RESULTS: The 3D model of Diplonema was compared with available crystal structures from Trypanosoma brucei, bacteriophage T4, and two archaeans. Interaction of DpRNL with ATP is predicted to involve double π-stacking, which has not been reported before in RNA ligases. This particular contact would shift the orientation of ATP and have considerable consequences on the interaction network of amino acids in the catalytic pocket. We postulate that certain canonical amino acids assume different functional roles in DpRNL compared to structurally homologous residues in other RNA ligases 2, a reassignment indicative of constructive neutral evolution. Finally, both structure comparison and phylogenetic analysis show that DpRNL is not specifically related to RNA ligases from trypanosomes, suggesting a unique adaptation of the latter for RNA editing, after the split of diplonemids and kinetoplastids. CONCLUSION: Homology modeling and molecular dynamics simulations strongly suggest that DpRNL is an RNA ligase 2. The predicted innovative reshaping of DpRNL's catalytic pocket is worthwhile to be tested experimentally.
[Mh] Termos MeSH primário: Euglenozoários/genética
Proteínas de Protozoários/química
Proteínas de Protozoários/metabolismo
RNA Ligase (ATP)/química
RNA Ligase (ATP)/metabolismo
[Mh] Termos MeSH secundário: Trifosfato de Adenosina/metabolismo
Domínio Catalítico
Euglenozoários/química
Euglenozoários/enzimologia
Magnésio/metabolismo
Cadeias de Markov
Modelos Moleculares
Simulação de Acoplamento Molecular
Simulação de Dinâmica Molecular
Filogenia
Proteínas de Protozoários/genética
RNA Ligase (ATP)/genética
Homologia Estrutural de Proteína
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Protozoan Proteins); 8L70Q75FXE (Adenosine Triphosphate); EC 6.5.1.3 (RNA Ligase (ATP)); I38ZP9992A (Magnesium)
[Em] Mês de entrada:1601
[Cu] Atualização por classe:170220
[Lr] Data última revisão:
170220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:151010
[St] Status:MEDLINE
[do] DOI:10.1186/s12900-015-0046-0


  9 / 39 MEDLINE  
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[PMID]:26294177
[Au] Autor:Lukes J; Flegontova O; Horák A
[Ad] Endereço:Biology Centre, Institute of Parasitology, Czech Academy of Sciences and Faculty of Sciences, University of South Bohemia, 37005 Ceské Budejovice (Budweis), Czech Republic; Canadian Institute for Advanced Research, Toronto, ON M5G 1Z8, Canada.
[Ti] Título:Diplonemids.
[So] Source:Curr Biol;25(16):R702-4, 2015 Aug 17.
[Is] ISSN:1879-0445
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Lukes et al. introduce an enigmatic group of unicellular eukaryotes called the diplonemids, which according to recent surveys may be widespread in marine ecosystems.
[Mh] Termos MeSH primário: Biodiversidade
Euglenozoários/classificação
Euglenozoários/fisiologia
[Mh] Termos MeSH secundário: Organismos Aquáticos/classificação
Organismos Aquáticos/genética
Organismos Aquáticos/fisiologia
Euglenozoários/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1605
[Cu] Atualização por classe:150821
[Lr] Data última revisão:
150821
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150822
[St] Status:MEDLINE


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[PMID]:25655777
[Au] Autor:Garrison CE; Bochdansky AB
[Ad] Endereço:Old Dominion University, Department of Ocean, Earth and Atmospheric Sciences, 4600 Elkhorn Ave., Norfolk, VA 23529, USA. Electronic address: cgarr017@gmail.com.
[Ti] Título:A simple separation method for downstream biochemical analysis of aquatic microbes.
[So] Source:J Microbiol Methods;111:78-86, 2015 Apr.
[Is] ISSN:1872-8359
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:In order to study the chemical composition of aquatic microbes it is necessary to obtain completely separated fractions of subpopulations. Size separation by filtration is usually unsuccessful because the smaller group of organisms contaminates the larger fractions due to being trapped on filter surfaces of nominally much larger pore sizes. Here we demonstrate that a simple sucrose density separation method allowed us to separate microorganisms of even subtle size differences and to determine their bulk biochemical composition (proteins, polysaccharides+nucleic acids, and lipids). Both autotrophs and heterotrophs (through anaplerotic pathways) were labeled with (14)C-bicarbonate for biochemical fractionation. We provided proof of concept that eukaryotic microbes could be cleanly separated from prokaryotes in cultures and in field samples, enabling detection of differences in their biochemical makeup. We explored methodological issues regarding separation mechanisms, fixation, and pre-concentration via tangential flow filtration of oligotrophic marine waters where abundances of microorganisms are comparably low. By selecting an appropriate centrifugal force, two processes (i.e., isopycnal and rate-zonal separation) can be exploited simultaneously resulting in finely-separated density fractions, which also resulted in size separation. Future applications of this method include exploration of the stoichiometric, biochemical and genetic differences among subpopulations of microbes in a wide variety of aquatic environments.
[Mh] Termos MeSH primário: Fracionamento Químico/métodos
Escherichia coli/química
Escherichia coli/isolamento & purificação
Euglenozoários/química
Euglenozoários/isolamento & purificação
Estramenópilas/química
Estramenópilas/isolamento & purificação
Microbiologia da Água
[Mh] Termos MeSH secundário: Organismos Aquáticos/química
Organismos Aquáticos/isolamento & purificação
Centrifugação com Gradiente de Concentração
Diatomáceas/química
Diatomáceas/isolamento & purificação
Filtração/métodos
Lipídeos/isolamento & purificação
Ácidos Nucleicos/isolamento & purificação
Polissacarídeos Bacterianos/isolamento & purificação
Proteínas/isolamento & purificação
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, U.S. GOV'T, NON-P.H.S.
[Nm] Nome de substância:
0 (Lipids); 0 (Nucleic Acids); 0 (Polysaccharides, Bacterial); 0 (Proteins)
[Em] Mês de entrada:1512
[Cu] Atualização por classe:150310
[Lr] Data última revisão:
150310
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150207
[St] Status:MEDLINE



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