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1.
Arch Microbiol ; 203(6): 3633-3639, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33993324

RESUMO

A Gram-stain-negative, rod-shaped, aerobic and non-motile bacterium, designated P2-65T, was isolated from Moonsan stream water in the Republic of Korea. The temperature, NaCl concentration and pH ranges for growth of strain P2-65T were 10-37 °C, 0.0-3.0% (w/v) and 6.5-8.5 with optimum growth at 25-30 °C, 0.0-1.0% and 7.0-7.5, respectively. Comparison of 16S rRNA gene sequence showed that strain P2-65T was closely related to Flavobacterium cauense (95.4%) and Flavobacterium cheniae (95.3%). The major fatty acids were iso-C15:0, iso C17:0 3-OH, summed feature 3 (C16:1ω7c and/or C16:1ω6c), summed feature 9 (iso-C17:1 ω9c and/or 10-methyl C16:0) and iso-C15:0 3-OH. The predominant respiratory quinone was menaquinone-6 (MK-6). The major polar lipids detected in the strain were phosphatidylethanolamine, one aminophospholipid, one unidentified aminolipid and one unidentified polar lipid. The G + C content of the genomic DNA was 39.7%. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values for strain P2-65T with closely related Flavobacterium species were below 74.8% and 20%, respectively. Based on polyphasic features, strain P2-65T is considered to represent a novel species of the genus Flavobacterium, for which the name Flavobacterium inviolabile sp. nov. is proposed. The type strain is P2-65T (= KCTC 62055T = NBRC 112953T).


Assuntos
Flavobacterium/classificação , Microbiologia da Água , Ácidos Graxos/análise , Flavobacterium/química , Flavobacterium/genética , Fosfatidiletanolaminas/análise
2.
Meat Sci ; 171: 108271, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32827804

RESUMO

Ruminant meats contain ester and ether-linked phosphatidylcholines-(PC) and phosphatidylethanolamines-(PE) enriched with ω3 and ω6 polyunsaturated fatty acids-(PUFA) essential for human health and nutrition. Oxidative degradation of these lipids during grilling compromises meat quality and safety. The effect of marinades containing unfiltered session ales, herbs and spices on these lipids in grilled beef and moose meat was investigated in current study. Marination preserved (P < 0.05) ester and ether linked PUFA-enriched PC and PE in moose, and PUFA-enriched ether PC and diacyl PE in beef against oxidative degradation. Furthermore, India ale-based marinated meats retained higher (P < 0.05) PUFA-enriched lysophosphatidylcholines-(LPC) and lysophosphatidylethanolamines-(LPE) compared to Wheat ale-based marinated meats. The preserved PUFA-enriched lipids were positively correlated with phenolics, oxygenated terpenes, and antioxidants present in the marinades, and negatively correlated to oxidation status. These findings appear to suggest that unfiltered beer-based marination could be a useful precooking technique to increase dietary access and consumption of essential fatty acids while preserving grilled meat nutritional quality and safety.


Assuntos
Culinária/métodos , Fosfatidilcolinas/análise , Fosfatidiletanolaminas/análise , Carne Vermelha/análise , Animais , Cerveja , Bovinos , Cervos , Ácidos Graxos Insaturados/análise
3.
Anal Bioanal Chem ; 413(2): 445-453, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33130974

RESUMO

Changes in the membrane composition of sub-populations of cells can influence different properties with importance to tumour growth, metastasis and treatment efficacy. In this study, we use correlated fluorescence microscopy and ToF-SIMS with C60+ and (CO2)6k+ ion beams to identify and characterise sub-populations of cells based on successful transfection leading to over-expression of CCTδ, a component of the multi-subunit molecular chaperone named chaperonin-containing tailless complex polypeptide 1 (CCT). CCT has been linked to increased cell growth and proliferation and is known to affect cell morphology but corresponding changes in lipid composition of the membrane have not been measured until now. Multivariate analysis of the surface mass spectra from single cells, focused on the intact lipid ions, indicates an enrichment of phosphatidylethanolamine species in the transfected cells. While the lipid changes in this case are driven by the structural changes in the protein cytoskeleton, the consequence of phosphatidylethanolamine enrichment may have additional implications in cancer such as increased membrane fluidity, increased motility and an ability to adapt to a depletion of unsaturated lipids during cancer cell proliferation. This study demonstrates a successful fluorescence microscopy-guided cell by cell membrane lipid analysis with broad application to biological investigation.Graphical abstract.


Assuntos
Microscopia de Fluorescência/métodos , Chaperonas Moleculares/análise , Neoplasias/metabolismo , Fosfatidiletanolaminas/análise , Espectrometria de Massa de Íon Secundário/métodos , Animais , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Proliferação de Células , Citoesqueleto/metabolismo , Ouro , Proteínas de Fluorescência Verde/metabolismo , Íons , Lipídeos/química , Melanoma Experimental , Camundongos , Análise Multivariada , Análise de Componente Principal
4.
Biomolecules ; 10(7)2020 07 18.
Artigo em Inglês | MEDLINE | ID: mdl-32708453

RESUMO

The structures of bioactive polar lipids (PLs) of Irish ale with potent antithrombotic and cardioprotective properties were elucidated. Ale PL was fractionated by preparative thin layer chromatography (TLC) into subclasses, and their antithrombotic effect was assessed against human platelet aggregation induced by the pro-inflammatory mediator, platelet-activating factor (PAF). The fatty acid content and the overall structures of ale PL were elucidated by liquid chromatography mass spectrometry (LC-MS). Phosphatidylcholines (PC) and molecules of the sphingomyelin (SM) family exhibited the strongest anti-PAF effects, followed by phosphatidylethanolamines (PE). PC contained higher amounts of omega-3 polyunsaturated fatty acids (n-3 PUFA) and thus the lowest n-6/n-3 ratio. Bioactive diacyl and alkyl-acyl PC and PE molecules bearing n-3 PUFA at their sn-2 position, especially docosahexaenoic acid (DHA) and α-linolenic acid (ALA) but mostly oleic acid (OA), were identified in both PC and PE subclasses. Eicosapentaenoic acid (EPA) was present only in bioactive PC molecules and not in PE, explaining the lower anti-PAF effects of PE. Bioactive sphingolipid and glycolipid molecules with reported anti-inflammatory and anti-tumour properties, such as specific ceramides and glucosylcerebrosides with sphingosine, phytosphingosine and dihydrosphingosine bases but also specific monogalactodiglycerides and SM species bearing ALA at their sn-2 position, were identified in the SM subclass, providing a rational for its strong bioactivities against the PAF pathway. Further studies are required on the health benefits of bioactive PL from beer and brewery by-products.


Assuntos
Cerveja/análise , Fator de Ativação de Plaquetas/antagonistas & inibidores , Inibidores da Agregação Plaquetária/análise , Inibidores da Agregação Plaquetária/farmacologia , Ácidos Graxos Ômega-3/análise , Ácidos Graxos Ômega-3/farmacologia , Humanos , Fosfatidilcolinas/análise , Fosfatidilcolinas/farmacologia , Fosfatidiletanolaminas/análise , Fosfatidiletanolaminas/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Esfingomielinas/análise , Esfingomielinas/farmacologia
5.
Sci Rep ; 10(1): 10250, 2020 06 24.
Artigo em Inglês | MEDLINE | ID: mdl-32581232

RESUMO

Oncogenic drivers of progression of monoclonal gammopathy of undetermined significance (MGUS) to multiple myeloma (MM) such as c-MYC have downstream effects on intracellular metabolic pathways of clonal plasma cells (PCs). Thus, extracellular environments such as the bone marrow (BM) plasma likely have unique metabolite profiles that differ from patients with MGUS compared to MM. This study utilized an untargeted metabolite and targeted complex lipid profiling of BM plasma to identify significant differences in the relative metabolite levels between patients with MGUS and MM from an exploratory cohort. This was followed by verification of some of the metabolite differences of interest by targeted quantification of the metabolites using isotopic internal standards in the exploratory cohort as well as an independent validation cohort. Significant differences were noted in the amino acid profiles such as decreased branch chain amino acids (BCAAs) and increased catabolism of tryptophan to the active kynurenine metabolite 3-hydroxy-kynurenine between patients with MGUS and MM. A decrease in the total levels of complex lipids such as phosphatidylethanolamines (PE), lactosylceramides (LCER) and phosphatidylinositols (PI) were also detected in the BM plasma samples from MM compared to MGUS patients. Thus, metabolite and complex lipid profiling of the BM plasma identifies differences in levels of metabolites and lipids between patients with MGUS and MM. This may provide insight into the possible differences of the intracellular metabolic pathways of their clonal PCs.


Assuntos
Metabolômica/métodos , Gamopatia Monoclonal de Significância Indeterminada/diagnóstico , Mieloma Múltiplo/diagnóstico , Plasmócitos/metabolismo , Aminoácidos de Cadeia Ramificada/análise , Diagnóstico Diferencial , Humanos , Cinurenina/análise , Lactosilceramidas/análise , Lipidômica/métodos , Gamopatia Monoclonal de Significância Indeterminada/metabolismo , Mieloma Múltiplo/sangue , Mieloma Múltiplo/metabolismo , Fosfatidiletanolaminas/análise , Fosfatidilinositóis/análise , Estudos Prospectivos
6.
Lipids ; 55(3): 271-278, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32255515

RESUMO

Nonalcoholic fatty liver disease (NAFLD) is associated with an imbalance in fatty acid composition and can progress from simple steatosis to steatohepatitis, liver cirrhosis, and hepatocellular carcinoma. Essential phospholipids (EPL), which contain high levels of 1,2-dilinoleoylphosphatidylcholine, can be used to treat NAFLD. Polyenylphosphatidylcholine (PPC) preparations are external, commercially available EPL products. The lipid composition of five commercially available PPC preparations, including Essentiale Forte, Fortifikat, Hepatoprotect Regenerator, Fortifikat Forte, and Esentin Forte were compared, the outcome of which may impact physician choice in the treatment of NAFLD. Following lipid extraction, a comparative analysis of key lipid content was performed using a QTRAP6500+ triple quadruple ion trap hybrid mass spectrometer (Sciex) in nanoelectrospray ionization mode. The glycerophospholipid composition of each PPC was determined, including levels of phosphatidylcholine (PtdCho), and phosphatidylethanolamine (PtdEtn) species, as well as PtdCho:PtdEtn ratio. Of the five preparations analyzed, Essentiale Forte contained the highest PtdCho levels (61.9 mol%) and lowest PtdEtn levels (4.9 mol%). PtdCho 36:4 levels, a polyunsaturated species of PtdCho, were highest in Esentin Forte (39.3 mol%) and Essentiale Forte (38.3 mol%) compared with other PPCs (28.7-35.8 mol%). Levels of lysophosphatidylcholine, phosphatidylinositol, phosphatidic acid, and phosphatidylglycerol were low in all five preparations. Lipid composition was consistent between the preparations. The high PtdCho:PtdEtn ratio composition of Essentiale Forte compared with the other PPC analyzed, as well as the presence of polyunsaturated fatty acids, suggest it could be the most clinically beneficial commercially available hepatoprotective product in the treatment of NAFLD.


Assuntos
Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Preparações Farmacêuticas/química , Fosfatidilcolinas/análise , Fosfatidiletanolaminas/análise , Tomada de Decisão Clínica , Humanos , Espectrometria de Massas
7.
Arch Microbiol ; 202(6): 1295-1300, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32125451

RESUMO

A novel Gram-stain-negative, orange, rod or curved rod, facultatively anaerobic, gliding bacterial strain, designated strain W255T, was isolated from Xiaoshi Island, Weihai, China. The strain W255T grows optimally at a 28 °C, pH 7.5, and 3.0% (w/v) NaCl environment. Its colonies are circular, orange, non-transparent, smooth, and approximately 0.2-0.8 mm in diameter, after being cultured for 72 h on marine agar 2216. Cells of the strain W255T are 0.3-0.8 µm wide and 1.0-4.0 µm long, catalase-positive and oxidase-positive. The major cellular fatty acids are iso-C15:0, iso-C15:1 G, and iso-C15:0 3-OH. The sole respiratory quinone is MK-6. The major polar lipids include phosphatidylethanolamine, one unidentified amino lipid, one amino glycolipid, and two unidentified lipids (L1 and L2). The phylogenetic analysis based on 16S rRNA gene sequences indicated that strain W255T has the highest similarities with the type strain of the type species of the genus Seonamhaeicola, S. aphaedonensis KCTC 32578T (97.2%), and moderate with 'S. acroporae' KCTC 62713T (96.5%), S. algicola Gy8T (95.4%) and S. marinus B011T (94.5%). The ANI and dDDH values between strain W255T and S. aphaedonensis KCTC 32578T are 86.6% and 31.3%, respectively. The genomic DNA G + C content is 33.5 mol%. On the basis of gene annotation, it was observed that strain W255T have the abilities of nitrate reduction and utilizing various carbon sources, suggesting that this strain might be an important participant in the nitrogen cycle and carbon cycle in the marine environment. Based on the phenotypic, chemotaxonomic and phylogenetic analysis, strain W255T has been considered as a novel species of the genus Seonamhaeicola, for which the name Seonamhaeicola sediminis sp. nov. is proposed. The type strain is W255T (= MCCC 1H00377T = KCTC 72085T).


Assuntos
Flavobacteriaceae/classificação , Sedimentos Geológicos/microbiologia , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/análise , Flavobacteriaceae/genética , Lipídeos/análise , Fosfatidiletanolaminas/análise , RNA Ribossômico 16S/genética , Água do Mar/microbiologia , Especificidade da Espécie , Vitamina K 2/química
8.
Food Chem ; 313: 125822, 2020 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-31931419

RESUMO

Concentrated pomegranate peel extract (CPE) was supplemented to ewes, and milk yield and fat content-fatty acid (FA) and phospholipid (PL) composition-were monitored. CPE-fed ewes had higher milk yield, and fat, protein and lactose contents than controls. Milk PL content-20% higher in the CPE-supplemented group-was regulated by treatment and not by total fat content; milk phosphatidylethanolamine and phosphatidylcholine increased by 22 and 26%, respectively, in CPE-supplemented vs. control ewes. Milk saturated FA concentration was higher, and total polyunsaturated and monounsaturated FA content lower in the CPE vs. control group, regardless of milk total fat content. CPE supplementation increased milk antioxidant capacity, suggesting antioxidant transfer from dietary source to milk, increasing stability and nutritive value. Our study provides first evidence for milk quality improvement in terms of antioxidants and PL enrichment without compromising total milk fat, suggesting strategies to improve dairy animals' milk composition without compromising total production.


Assuntos
Antioxidantes/metabolismo , Dieta , Leite/química , Punica granatum/química , Ração Animal/análise , Animais , Antioxidantes/química , Dieta/veterinária , Ácidos Graxos Monoinsaturados/análise , Feminino , Lactação , Leite/metabolismo , Valor Nutritivo , Fosfatidilcolinas/análise , Fosfatidiletanolaminas/análise , Punica granatum/metabolismo , Ovinos
9.
J Microbiol ; 57(12): 1079-1085, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31758394

RESUMO

A yellow pigmented, Gram-stain-negative, strictly aerobic, rod-shaped, motile by means of gliding, catalase and oxidase positive bacterium, designated strain DS2-AT, was isolated from soil. Growth was observed at 4-32°C (optimum, 28°C), pH 6-9 (optimum, 7.0), and with 0-0.25% (w/v) NaCl (optimum, 0%). Phylogenetic analysis of 16S rRNA gene sequence revealed that strain DS2-AT belonged to the genus Flavobacterium and was most closely related to Flavobacterium aquatile LMG 4008T (96.4%), Flavobacterium terrae DSM 18829T (95.6%), Flavobacterium vireti THG-SM1T (95.5%), Flavobacterium inkyongense IMCC27201T (95.4%), Flavobacterium brevivitae TTM-43T (95.2%), and Flavobacterium cucumis DSM 18830T (95.2%). Strain DS2-AT produces flexirubin-type pigments. The major fatty acids were iso-C15:0, iso-C17:0 3-OH, and iso-C15:0 3-OH. The major respiratory quinone was identified as menaquinone-6. The major polar lipid was found to be phosphatidylethanolamine. The average nucleotide identity values between strain DS2-AT and selected taxa, F. aquatile LMG 4008T, F. terrae DSM 18829T, and F. cucumis DSM 18830T, were 72, 72.7, and 71.6%, respectively. The draft genome of strain DS2-AT has a number of 14 contigs, scaffold N50 of 476,310 bp and a total size of 3,563,867 bp. Additionally, strain DS2-AT contains 3,127 of gene, 41 of tRNA, 6 of rRNA, and 3 of ncRNA. The DNA G + C content of stain DS2-AT was 40.7 mol%. Based on phylogenetic and phenotypic analyses, strain DS2-AT is considered as a novel species of the genus Flavobacterium, for which the name Flavobacterium humi sp. nov., (type strain DS2-AT = KACC 19715T = JCM 32786T) has been proposed.


Assuntos
Flavobacterium/classificação , Flavobacterium/isolamento & purificação , Flavobacterium/metabolismo , Filogenia , Pigmentos Biológicos/metabolismo , Polienos/metabolismo , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/análise , Flavobacterium/genética , Hibridização de Ácido Nucleico , Fosfatidiletanolaminas/análise , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Solo , Vitamina K 2/análogos & derivados , Vitamina K 2/análise
10.
J Phys Chem B ; 123(49): 10433-10440, 2019 12 12.
Artigo em Inglês | MEDLINE | ID: mdl-31729230

RESUMO

Model membranes are a valuable tool to investigate the mechanism of interaction between antibiotic compounds and bacterial membranes. However, the development of supported lipid bilayer (SLB) models for Gram-negative and Gram-positive bacteria has been challenging because of the high charge and spontaneous curvature of the lipids that make up these membranes. Here we describe a method for preparing mimetic Gram-negative inner membrane and Gram-positive membrane SLBs, including asymmetric SLBs (asy-SLBs) that contain a fluorescent tracer only in the upper leaflet of the membrane. We quantified the dynamics of the lipids in these membranes with fluorescence correlation spectroscopy (FCS) and found that lipid diffusion is slower in Gram-negative SLBs/asySLBs than in Gram-positive SLBs/asySLBs. Peptide binding to these membranes was also characterized using colistin, a Gram-negative specific antibiotic. Interactions between colistin and membrane lipids phosphatidylethanolamine (PE) or cardiolipin (TOCL) were probed with pulsed-interleaved excitation fluorescence cross-correlation spectroscopy (PIE-FCCS). Overall, our data provide unique insight into the diffusion dynamics of lipids in Gram-negative and Gram-positive membranes as well as a novel platform for investigating the mechanism of interaction between antibiotic peptides and bacterial membrane lipids.


Assuntos
Cardiolipinas/análise , Bactérias Gram-Negativas/química , Bactérias Gram-Positivas/química , Bicamadas Lipídicas/química , Peptídeos/química , Fosfatidiletanolaminas/análise , Sítios de Ligação , Espectrometria de Fluorescência
11.
J Microbiol ; 57(12): 1065-1072, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31562606

RESUMO

A yellow pigmented, Gram-stain-negative, aerobic bacterium designated A5.7T was studied to evaluate the taxonomic position following the modern polyphasic approach. The strain was isolated from sediments near Zhairuo Island, which is situated in the East China Sea. Cells were non-spore forming rods without flagella but showed motility by gliding. Growth was observed at 15-35°C (optimum 28°C), pH 6.0-9.0 (optimum pH 6.5) and 0-2% (w/v) NaCl (optimum 0-0.5%) in LB broth. The major respiratory quinone of A5.7T was menaquinone 6. The major polar lipid of A5.7T was phosphatidylethanolamine and the predominant fatty acids (> 5%) were iso-C15:0, iso-C17:0 3-OH, C15:1ω6c, iso-C15:0 3-OH, iso-C15:1 G, summed feature 3 (C16:1ω7c and/or C16:1ω6c) and summed feature 9 (iso-C17:1ω9c and/or C16:010-methyl). Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolate belongs to the genus Flavobacterium and shares the highest sequence similarities with Flavobacterium sharifuzzamanii A7.6T (98.5%), Flavobacterium tistrianum GB 56.1T (98.3%), Flavobacterium nitrogenifigens NXU-44T (97.8%), Flavobacterium anhuiense D3T (97.6%), Flavobacterium ginsenosidimutans THG 01T (97.6%), and Flavobacterium foetidum CJ42T (97.6%). Digital DNA-DNA hybridization and average nucleotide identity values between the strain and its closest phylogenetic neighbors showed the ranges from 19.6 to 34.1% and 73.7 to 87.9%, respectively. Therefore, based on polyphasic characteristics, strain A5.7T represents a novel species of the genus Flavobacterium for which the name Flavobacterium zhairuonensis sp. nov. is proposed. The type strain is A5.7T (= KCTC 62406T = MCCC 1K03494T).


Assuntos
Flavobacterium/classificação , Flavobacterium/isolamento & purificação , Sedimentos Geológicos/microbiologia , Filogenia , Água do Mar/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/análise , Flavobacterium/genética , Flavobacterium/fisiologia , Genoma Bacteriano/genética , Concentração de Íons de Hidrogênio , Hibridização de Ácido Nucleico , Fosfatidiletanolaminas/análise , Quinonas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Cloreto de Sódio , Temperatura , Vitamina K 2/análogos & derivados , Vitamina K 2/análise
12.
Arch Microbiol ; 201(8): 1119-1127, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31165177

RESUMO

Strain KYPW7T, isolated from the Funglin Stream in Taiwan, was characterized using a polyphasic taxonomy approach. Cells of strain KYPW7T were Gram-stain-negative, aerobic, non-spore forming, non-motile rods and formed white colonies. Growth occurred at 15-30 °C (optimum 25 °C), at pH 6-8 (optimum pH 6.5) and with 0-1% NaCl (optimum 0%). Phylogenetic analyses based on 16S rRNA and coding sequences of 92 protein clusters showed that strain KYPW7T represents a novel genus in the family Flavobacteriaceae. The 16S rRNA gene sequence of strain KYPW7T was related to the species of the genera Chryseobacterium (91.8-96.0% sequence similarity), Bergeyella (95.1-95.8%), Cloacibacterium (94.5-95.7%), Daejeonia (95.6%) and Riemerella (94.0-95.0%). Strain KYPW7T showed less than 72% average nucleotide identity and less than 24% digital DNA-DNA hybridization identity compared to the type strains of related genera within the family Flavobacteriaceae. The predominant fatty acids were iso-C15:0 and iso-C17:0 3-OH. The major isoprenoid quinone was MK-6 and the DNA G + C content was 36.8 mol%. The polar lipids had phosphatidylethanolamine, three uncharacterized aminophospholipids and an uncharacterized phospholipid. The polyamines contained homospermidine, putrescine and spermidine. On the basis of the genotypic and phenotypic data, strain KYPW7T represents a novel species of a new genus in the family Flavobacteriaceae, for which the name Amniculibacterium aquaticum gen. nov., sp. nov. is proposed. The type strain is KYPW7T (= BCRC 81123T = LMG 30598T = KCTC 62512T).


Assuntos
Flavobacteriaceae , Rios/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases/genética , DNA Bacteriano/genética , Ácidos Graxos/análise , Flavobacteriaceae/classificação , Flavobacteriaceae/genética , Flavobacteriaceae/isolamento & purificação , Hibridização de Ácido Nucleico , Fosfatidiletanolaminas/análise , Fosfolipídeos/análise , Filogenia , Poliaminas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Taiwan
13.
Arch Microbiol ; 201(8): 1053-1060, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31115641

RESUMO

Strain HPM-16T, isolated from seawater, was characterized using a polyphasic taxonomy approach. Phylogenetic analyses based on 16S rRNA gene sequences and coding sequences of an up-to-date bacterial core gene set (92 protein clusters) indicated that strain HPM-16T formed a phylogenetic lineage in the genus Neptunomonas. Strain HPM-16T was most closely related to Neptunomonas concharum LHW37T with 16S rRNA gene sequence similarity of 96.7%. Cells were Gram-stain negative, facultatively anaerobic, motile by means of a single polar flagellum, rod-shaped and formed white colonies. Optimal growth occurred at 30-35 °C, pH 6.5-8, and in the presence of 2-5% NaCl. C18:1ω7c and summed feature 3 (C16:1ω7c and/or C16:1ω6c) were the predominant fatty acids. The only isoprenoid quinone was Q-8. The polar lipid profile revealed the presence of phosphatidylethanolamine, phosphatidylglycerol and several uncharacterized lipids. The major polyamines were putrescine and spermidine. The draft genome was approximately 3.68 Mb in size with a G + C content of 50.5 mol%. Differential phenotypic properties, together with the phylogenetic inference, demonstrate that strain HPM-16T should be classified as a novel species of the genus Neptunomonas, for which the name Neptunomonas marina sp. nov. is presented. The type strain is HPM-16T (= BCRC 80980T = LMG 29560T = KCTC 52235T).


Assuntos
Oceanospirillaceae , Água do Mar/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases/genética , DNA Bacteriano/genética , Ácidos Graxos/análise , Oceanospirillaceae/classificação , Oceanospirillaceae/genética , Oceanospirillaceae/isolamento & purificação , Fosfatidiletanolaminas/análise , Fosfatidilgliceróis/análise , Fosfolipídeos/análise , Filogenia , Quinonas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
14.
Proteomics ; 19(18): e1800297, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30790445

RESUMO

The use of ultra performance liquid chromatography coupled to data independent tandem mass spectrometry with traveling wave ion mobility for detection and structural identification of ether-linked glycerophosphoethanolamine is described. The experimental design generates 4D data (chromatographic retention time, precursor accurate mass, drift time with associated calculated collisional cross-section, and time-aligned accurate mass diagnostic product ions) for each ionization mode. Confident structure identification depends on satisfying 4D data confirmation in both positive and negative ion mode. Using this methodology, a number of ether-linked glycerophosphoethanolamine lipids are structurally elucidated from mouse brain lysosomes. It is further determined that several ether-linked glycerophosphoethanolamine structures are differentially abundant between lysosomes isolated from mouse cortex following traumatic brain injury as compared to that of sham animals. The combined effort of aligning multi-dimensional mass spectrometry data with a well-defined traumatic brain injury model lays the foundation for gaining mechanistic insight in the role lysosomal membrane damage plays in neuronal cell death following brain injury.


Assuntos
Lesões Encefálicas Traumáticas/metabolismo , Córtex Cerebral/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Lisossomos/metabolismo , Fosfatidiletanolaminas/química , Fosfatidiletanolaminas/metabolismo , Espectrometria de Massas por Ionização por Electrospray/métodos , Animais , Lesões Encefálicas Traumáticas/patologia , Modelos Animais de Doenças , Éteres/química , Camundongos , Fosfatidiletanolaminas/análise
15.
J Microbiol ; 57(2): 122-126, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30706341

RESUMO

A yellow-colored bacterium with gliding motility, strain KIS68-18T, was isolated from a soil sample at Bijin Island in Tongyeong city, Republic of Korea. The cells were strictly aerobic, Gram-staining-negative, non-spore-forming, and rod-shaped. The strain grew at the range of 10-35°C (optimum, 25-30°C), pH 5.5-8.0 (optimum, 6.0-7.5), and 0-0.5% (w/v) NaCl. A phylogenetic analysis based on 16S rRNA gene sequences revealed that strain KIS68-18T was closely related to Chryseolinea serpens DSM 24574T (98.9%) and had low sequence similarities (below 92.6%) with other members of the family 'Cytophagaceae' in the phylum Bacteroidetes. The major respiratory quinone system was MK-7 and the predominant cellular fatty acids were C16:1ω5c (38.8%), iso-C15:0 (18.5%), and summed feature 3 (C16:1ω7c and/or C16:1ω6c, 10.6%). The polar lipids consisted of phosphatidylethanolamine, one unidentified phospholipid, three unidentified aminophospholipids, two unidentified aminolipids, and five unidentified lipids. The DNA G + C content was 50.9%. Based on the phylogenetic, physiological, and chemotaxonomic data, stain KIS68-18T represents a novel species of the genus Chryseolinea, for which the name Chryseolinea soli sp. nov. is proposed. The type strain of Chryseolinea soli is KIS68-18T (= KACC 17327T = NBRC 113100T).


Assuntos
Bacteroidetes/classificação , Bacteroidetes/isolamento & purificação , Filogenia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Bacteroidetes/genética , Bacteroidetes/fisiologia , Composição de Bases , Benzoquinonas/análise , Cytophagaceae/classificação , Cytophagaceae/genética , DNA Bacteriano/genética , Ácidos Graxos/análise , Fosfatidiletanolaminas/análise , Fosfolipídeos/análise , Pigmentação , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Solo , Especificidade da Espécie , Sequenciamento Completo do Genoma
16.
Chem Res Toxicol ; 32(4): 638-644, 2019 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-30735032

RESUMO

Benzyl isothiocyanate (BITC), a dietary isothiocyanate (ITC) derived from cruciferous vegetables, has anticancer properties. It is believed that the ITC moiety (-N═C═S) that reacts predominantly with thiol compounds plays a central role in triggering the activities resulting from these properties. Recent studies have demonstrated that ITCs also covalently modify amino moieties in a protein. In this study, we examined the chemical reaction between BITC and the aminophospholipid, phosphatidylethanolamine (PE), in the cell membrane or lipoprotein particle. To detect the BITC-modified PE, the bond between ethanolamine (EA) and phosphatidic acid in PE was cleaved using phospholipase D to form the BITC-EA adduct, which was then measured. BITC-EA was detected from the BITC-treated unilamellar liposome and low-density lipoprotein even with only a few micromoles of BITC treatment, suggesting that BITC might react with not only a thiol/amino group of a protein but also an amino moiety of an aminophospholipid. Moreover, after incorporating BITC-PE included in the liposomes into the cultured cells or after direct exposure of BITC to the cells, free BITC-EA was excreted and accumulated in the medium in a time-dependent manner. It indicates that an intracellular enzyme catalyzes the cleavage of BITC-PE to produce BITC-EA. Because the ITC-amine adduct is stable, the ITC-EA adduct could be a promising indicator of ITC exposure in vivo.


Assuntos
Etanolamina/metabolismo , Isotiocianatos/metabolismo , Fosfatidiletanolaminas/metabolismo , Animais , Etanolamina/análise , Isotiocianatos/análise , Lipossomos/química , Lipossomos/metabolismo , Camundongos , Estrutura Molecular , Fosfatidiletanolaminas/análise , Células RAW 264.7
17.
Clin Neuroradiol ; 29(1): 27-36, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28983683

RESUMO

PURPOSE: Previous ex vivo spectroscopic data from tissue samples revealed differences in phospholipid metabolites between isocitrate dehydrogenase mutated (IDHmut) and IDH wildtype (IDHwt) gliomas. We investigated whether these changes can be found in vivo using 1H-decoupled 31P magnetic resonance spectroscopic imaging (MRSI) with 3D chemical shift imaging (CSI) at 3 T in patients with low and high-grade gliomas. METHODS: The study included 33 prospectively enrolled, mostly untreated patients who met spectral quality criteria according to the World Health Organization (WHO II n = 7, WHO III n = 17, WHO IV n = 9; 25 patients IDHmut, 8 patients IDHwt). The MRSI protocol included 1H decoupled 31P MRSI with 3D CSI (3D 31P CSI), 2D 1H CSI and a 1H single voxel spectroscopy sequence (TE 30 ms) from the tumor area. For 1H MRS, absolute metabolite concentration values were calculated (phantom replacement method). For 31P MRS, metabolite intensity ratios were calculated for the choline (C) and ethanolamine (E)-containing metabolites. RESULTS: In our patient cohort we did not find significant differences for the ratio of phosphocholine (PC) and phosphoethanolamine (PE), PC/PE, (p = 0.24) for IDHmut compared to IDHwt gliomas. Furthermore, we found no elevated ratios of glycerophosphocholine (GPC) and glycerophosphoethanolamine (GPE), GPC/GPE, (p = 0.68) or GPC/PE (p = 0.12) for IDHmut gliomas. Even the ratio (PC+GPC)/(PE+GPE) showed no significant differences with respect to mutation status (p = 0.16). Nonetheless, changes related to tumor grade regarding intracellular pH (pHi) and phospholipid metabolism as well as absolute metabolite concentrations of co-registered 2D 1H CSI data for tumor and control tissue showed the anticipated results. CONCLUSION: Using 3D-CSI data acquisition, in vivo 31P MR spectroscopic measurement of phospholipid metabolites could not distinguish between IDHmut and IDHwt.


Assuntos
Biomarcadores Tumorais/genética , Neoplasias Encefálicas/metabolismo , Glioma/metabolismo , Isocitrato Desidrogenase/genética , Espectroscopia de Ressonância Magnética/métodos , Adulto , Idoso , Análise de Variância , Astrocitoma/enzimologia , Astrocitoma/genética , Astrocitoma/patologia , Astrocitoma/terapia , Biomarcadores Tumorais/metabolismo , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/terapia , Diagnóstico Diferencial , Etanolaminas/análise , Etanolaminas/metabolismo , Feminino , Glioblastoma/enzimologia , Glioblastoma/genética , Glioblastoma/patologia , Glioblastoma/terapia , Glioma/genética , Glioma/patologia , Glioma/terapia , Glutaratos/análise , Glutaratos/metabolismo , Glicerilfosforilcolina/análise , Humanos , Hidrogênio , Isocitrato Desidrogenase/metabolismo , Isoenzimas/análise , Isoenzimas/metabolismo , Masculino , Pessoa de Meia-Idade , Mutação , Gradação de Tumores , Oligodendroglioma/enzimologia , Oligodendroglioma/genética , Oligodendroglioma/patologia , Oligodendroglioma/terapia , Fosfatidiletanolaminas/análise , Fosfatidiletanolaminas/metabolismo , Isótopos de Fósforo , Fosforilcolina/análise , Fosforilcolina/metabolismo , Estudos Prospectivos , Carga Tumoral
18.
Arch Microbiol ; 201(1): 81-86, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30238204

RESUMO

A novel yellow-colored, Gram-stain-negative, aerobic, non-motile, catalase- and oxidase-positive, and rod-shaped psychrotolerant bacterium, designated strain PLR-18-3T, was isolated from Arctic soil and was subjected to polyphasic taxonomic study. Cells were able to grow at 0-30 °C, pH 6.0-10.5, and 0-3.0% (w/v) NaCl concentration. Based on the 16S rRNA gene sequence analysis, this Arctic strain belonged to the genus Flavobacterium, with the closest neighbor being Flavobacterium noncentrifugens R-HLS-17T (96.2% sequence similarity). The strain contained MK-6 as a sole respiratory quinone, phosphatidylethanolamine as the major polar lipid, and summed feature 3 (C16:1ω7c and/or C16:1ω6c), iso-C15:0, iso-C15:0 G, iso-C17:0 3-OH, iso-C15:0 3-OH, anteiso-C15:0, and summed feature 9 (iso-C17:1ω9c and/or C16:010-methyl) as the predominant fatty acids. The DNA G + C content was 37.9 mol%. On the basis of polyphasic data, strain PLR-18-3T represents a novel species of the genus Flavobacterium, for which the name Flavobacterium dasani sp. nov. is proposed. The type strain is PLR-18-3T (=KEMB 9005-713T=KACC 19627T=NBRC 113347T).


Assuntos
Flavobacterium , Regiões Árticas , Técnicas de Tipagem Bacteriana , Composição de Bases , Temperatura Baixa , DNA Bacteriano/genética , Ácidos Graxos/análise , Flavobacterium/classificação , Flavobacterium/genética , Flavobacterium/isolamento & purificação , Camada de Gelo/microbiologia , Fosfatidiletanolaminas/análise , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Solo , Microbiologia do Solo , Vitamina K 2/análise
19.
J Microbiol ; 57(2): 107-112, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30456756

RESUMO

The novel Gram-stain-negative, rod-shaped, aerobic bacterial strain DCR-13T was isolated from a native plant belonging to the genus Campanula on Dokdo, an island in the Republic of Korea. Comparative analysis of the 16S rRNA gene sequence indicated that this strain is closely related to Paraburkholderia peleae PP52-1T (98.43% 16S rRNA gene sequence similarity), Paraburkholderia oxyphila NBRC 105797T (98.42%), Paraburkholderia sacchari IPT 101T (98.28%), Paraburkholderia mimosarum NBRC 106338T (97.80%), Paraburkholderia denitrificans KIS30-44T (97.46%), and Paraburkholderia paradise WAT (97.45%). This analysis of the 16S rRNA gene sequence also suggested that DCR-13T and the six closely related strains formed a clade within the genus Paraburkholderia, but that DCR-13T was clearly separated from the established species. DCR-13T had ubiquinone 8 as its predominant respiratory quinone, and its genomic DNA G + C content was 63.9 mol%. The isolated strain grew at a pH of 6.0-8.0 (with an optimal pH of 6.5), 0-4% w/v NaCl (with an optimal level of 0%), and a temperature of 18-42°C (with an optimal temperature of 30°C). The predominant fatty acids were C16:0, summed feature 8 (C18:1ω7c/C18:1ω6c), C17:0 cyclo, C19:0 cyclo ω8c, summed feature 3 (C16:1ω6c/C16:1ω7c) and summed feature 2 (C12:0 aldehyde), and the major polar lipids were phosphatidylglycerol and phosphatidylethanolamine. On the basis of polyphasic evidence, it is proposed that strain DCR-13T (= KCTC 62811T = LMG 30889T) represents the type strain of a novel species, Paraburkholderia dokdonella sp. nov.


Assuntos
Burkholderiaceae/classificação , Burkholderiaceae/isolamento & purificação , Campanulaceae/microbiologia , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , Benzoquinonas , Burkholderiaceae/genética , Burkholderiaceae/fisiologia , DNA Bacteriano/análise , Ácidos Graxos/análise , Ilhas , Hibridização de Ácido Nucleico , Fosfatidiletanolaminas/análise , Fosfatidilgliceróis/análise , Fosfolipídeos/análise , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Microbiologia do Solo , Especificidade da Espécie , Temperatura , Ubiquinona
20.
Arch Microbiol ; 200(10): 1481-1486, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30159757

RESUMO

A novel Gram-reaction-negative, rod-shaped, non-motile bacterium, designated as strain G2-10T was isolated from effluent of a dairy manure treatment plant. Growth occurred at 20-40 °C (optimum at 25-30 °C), pH 7.0-8.0 (optimum at pH 7.0). The range of NaCl concentration for growth was between 0% and 3% (w/v) (optimum 0-1%, w/v). Comparison of 16S rRNA gene sequence indicated that strain G2-10T was moderately related to the type strains of Sphingobacterium nematocida M-SX103T and Sphingobacterium suaedae T47T with a pair-wise sequence similarity of 94.3% and 94.0%, respectively. The major fatty acid constituents of strain G2-10T were identified as iso-C15:0 (37.6%), summed feature 3 (consisting of C16:1ω7c and/or C16:1ω6c, 29.6%) and iso-C17:0 3-OH (15.2%). Phosphatidylethanolamine was the major polar lipids of strain G2-10T. Sphingophospholipids were present. The isoprenoid quinone was composed of only MK-7. The DNA G + C content of strain G2-10T was found to be 42.5 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain G2-10T represents a novel species within the genus Sphingobacterium, for which the name Sphingobacterium praediipecoris is proposed. The type strain is G2-10T (= KCTC 52880T = NBRC 112848T).


Assuntos
Sphingobacterium/classificação , Composição de Bases , DNA Bacteriano/química , Indústria de Laticínios , Ácidos Graxos/análise , Esterco , Fosfatidiletanolaminas/análise , Filogenia , RNA Ribossômico 16S/genética , Sphingobacterium/química , Sphingobacterium/genética , Sphingobacterium/isolamento & purificação
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