RESUMO
Two novel Gram-stain-positive, non-motile and non-spore-forming bacterial strains, designated J2M5T and J1M15T, were isolated from the gastrointestinal tract of a lake prawn Palaemon paucidens. Strain J2M5T was an obligately aerobic bacterium that formed milky-coloured colonies and showed a rod-coccus cell cycle, while strain J1M15T was a facultatively aerobic bacterium that formed orangish-yellow-coloured colonies and showed rod-shaped cells. Strains J2M5T and J1M15T showed the highest 16S rRNA gene sequence similarity to Nocardioides ganghwensis JC2055T (98.63â%) and Tessaracoccus flavescens SST-39T (98.08â%), respectively. The whole-genome sequence of strain J2M5T was 4.52 Mbp in size and the genomic G+C content directly calculated from the genome sequence of strain J2M5T was 72.5âmol%. The whole-genome sequence of strain J1M15T was 3.20 Mbp in size and the genomic G+C content directly calculated from the genome sequence of strain J1M15T was 69.6molâ%. Strains J2M5T and J1M15T showed high OrthoANI similarity to N. ganghwensis JC2055T (83.6â%) and T. flavescens (77.2â%), respectively. We analysed the genome sequences of strains J2M5T and J1M15T in terms of carbohydrate-active enzymes, antibiotic resistance genes and virulence factor genes. Strains J2M5T and J1M15T contained MK-8 (H4) and MK-9 (H4) as the predominant respiratory quinones, respectively. The major polar lipids of both strains were phosphatidylglycerol and diphosphatidylglycerol. Additionally, strain J2M5T possessed phosphatidylcholine, phosphatidylserine and phosphatidylethanolamine. The cellular sugar components of strain J2M5T were ribose, mannose, glucose and galactose, and its cellular amino acid components were l-alanine and l-lysine. The cellular sugar components of strain J1M15T were rhamnose, ribose, mannose and glucose, and its cellular amino acid component was l-alanine. The major cellular fatty acids of strains J2M5T and J1M15T were iso-C16â: 0 and anteiso-C15â:â0, respectively. The multiple taxonomic analyses indicated that strains J2M5T and J1M15T represent novel species of the genus Nocardioides and Tessaracoccus, respectively. We propose the names Nocardioides palaemonis sp. nov. and Tessaracoccus palaemonis sp. nov. for strain J2M5T (=KCTC 49461T=CCUG 74767T) and strain J1M15T (=KCTC 49462T=CCUG 74766T), respectively.
Assuntos
Ácidos Graxos , Fosfolipídeos , Ácidos Graxos/química , Fosfolipídeos/química , Nocardioides , RNA Ribossômico 16S/genética , Lagos , Manose , Ribose , Análise de Sequência de DNA , Filogenia , Composição de Bases , DNA Bacteriano/genética , Técnicas de Tipagem Bacteriana , Trato Gastrointestinal , Alanina/genética , Aminoácidos , Glucose , Vitamina K 2/químicaRESUMO
A novel bacterium, designated strain JHSY0214T, was isolated from the gut of a Korean limpet, Cellana toreuma. Cells of strain JHSY0214T were Gram-stain-negative, strictly aerobic, yellow-pigmented, non-spore-forming, non-motile and showed a rod-coccus growth cycle. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain belonged to the genus Parasphingorhabdus, and was most closely related to Parasphingorhabdus litoris KCTC 12764T (98.71â%). Strain JHSY0214T had two fluoroquinolone-resistance genes and seven multidrug-resistance efflux pump genes, but did not have beta-lactamase genes and zinc resistance genes compared with P. litoris KCTC 12764T. Strain JHSY0214T grew optimally at 30 °C, pH 7.0 and in the presence of 2â% (w/v) NaCl. The predominant cellular fatty acids of strain JHSY0214T were summed feature 8 (C18â:â1 ω6c and/or C18â:â1 ω7c; 41.2â%), summed feature 3 (C16â:â1 ω7c and/or C16â:â1 ω6c; 21â%) and C16â:â0 (18.9â%). The major isoprenoid quinone was ubiquinone-10. The major polar lipids were sphingoglycolipid and phosphatidylethanolamine. The genomic DNA G+C content was 52.8 mol%. Based on phylogenetic, genotypic and phenotypic data, strain JHSY0214T represents a novel species of the genus Parasphingorhabdus, for which the name Parasphingorhabdus cellanae is proposed. The type strain is JHSY0214T (=KCTC 82387T=DSM 112279T).
Assuntos
Ácidos Graxos , Gastrópodes , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/químicaRESUMO
The LIM-homeodomain transcription factor Lmx1b plays a key role in body pattern formation during development. Although Lmx1b is essential for the normal development of multiple tissues, its regulatory mechanism in bone cells remains unclear. Here, we demonstrated that Lmx1b negatively regulates bone morphogenic protein 2 (BMP2)-induced osteoblast differentiation. Overexpressed Lmx1b in the osteoblast precursor cells inhibited alkaline phosphatase (ALP) activity and nodule formation, as well as the expression of osteoblast maker genes, including runt-related transcription factor 2 (Runx2), alkaline phosphatase (Alpl), bone sialoprotein (Ibsp), and osteocalcin (Bglap). Conversely, the knockdown of Lmx1b in the osteoblast precursors enhanced the osteoblast differentiation and function. Lmx1b physically interacted with and repressed the transcriptional activity of Runx2 by reducing the recruitment of Runx2 to the promoter region of its target genes. In vivo analysis of BMP2-induced ectopic bone formation revealed that the knockdown of Lmx1b promoted osteogenic differentiation and bone regeneration. Our data demonstrate that Lmx1b negatively regulates osteoblast differentiation and function through regulation of Runx2 and provides a molecular basis for therapeutic targets for bone diseases.
Assuntos
Subunidade alfa 1 de Fator de Ligação ao Core , Fatores de Transcrição , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Proteína Morfogenética Óssea 2/genética , Proteína Morfogenética Óssea 2/metabolismo , Diferenciação Celular/genética , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Osteoblastos/metabolismo , Osteogênese/genética , Fatores de Transcrição/metabolismoRESUMO
A polyphasic taxonomic approach was used to characterize two novel bacterial strains, designated as HDW11T and HDW19T, isolated from intestine samples of the dark diving beetle Hydrophilus acuminatus and the diving beetle Cybister lewisianus, respectively. Both isolates were Gram-stain-positive, facultatively anaerobic and non-motile. Strain HDW11T grew optimally at 30 °C, pH 8 and in the presence of 1% (w/v) NaCl. Strain HDW19T grew optimally at 25 °C, pH 7 and in the presence of 0.3% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences and genome sequences revealed that strain HDW11T is a member of the genus Brevilactibacter and is closely related to Brevilactibacter flavus VG341T [with 97.9% 16S rRNA sequence identity and 79.1% average nucleotide identity (ANI)], and that strain HDW19T belongs to the genus Weissella and is closely related to W. koreensis KCTC 3621T (with 98.9% 16S rRNA sequence identity and 79.5% ANI). The major cellular fatty acids of strains HDW11T and HDW19T were C18:1 ω9c and anteiso-C15:0, respectively. The sole respiratory quinone of strain HDW11T was MK-9 (H4). The major polar lipid components of strain HDW11T were diphosphatidylglycerol and phosphatidylglycerol, and the major polar lipid component of strain HDW19T was diphosphatidylglycerol. The genomic DNA G+C content of strains HDW11T and HDW19T were 72.1 and 37.2 mol%, respectively. The results of phylogenetic, phenotypic, chemotaxonomic and genotypic analyses suggest that strain HDW11T represents a novel species within the genus Brevilactibacter, and that strain HDW19T represents a novel species within the genus Weissella. We propose the name Brevilactibacter coleopterorum sp. nov. for strain HDW11T (=KACC 21335T=KCTC 49320T=JCM 33680T) and the name Weissella coleopterorum for strain HDW19T (=KACC 21347T=KCTC 43114T=JCM 33684T).
Assuntos
Besouros/microbiologia , Intestinos/microbiologia , Filogenia , Propionibacteriaceae/classificação , Weissella/classificação , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , Besouros/classificação , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , Propionibacteriaceae/isolamento & purificação , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Weissella/isolamento & purificaçãoRESUMO
A novel Gram-stain-negative, aerobic, rod-shaped bacterium with a single polar flagellum, designated strain 2T18T, was isolated from the gut of the freshwater mussel Anodonta arcaeformis collected in the Republic of Korea. Phylogenetic analyses based on 16S rRNA gene sequences showed that the strain belonged to the genus Chitinibacter. Strain 2T18T formed a monophyletic clade with Chitinibacter fontanus KCTC 42982T, C. tainanensis KACC 11706T and C. alvei KCTC 23839T, with sequence similarities of 98.5, 98.4 and 95.9â%, respectively. Strain 2T18T exhibited optimal growth at 30 °C, at pH 8 and with 0.5â% (w/v) NaCl. The major isoprenoid quinone was ubiquinone-8 (Q-8). The predominant fatty acids were summed feature 3 (C16â:â1 ω6c and/or C16â:â1 ω7c) and C16â:â0. The polar lipids comprised phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, one unidentified lipid, three unidentified phospholipids and two unidentified aminophospholipids. The G+C content of the genomic DNA was 50.6 mol%. The average nucleotide identity and digital DNA-DNA hybridization values between strains 2T18T and C. fontanus KCTC 42982T were below the thresholds used for the delineation of a novel species. Based on the phylogenetic, phenotypic, chemotaxonomic and genotypic characteristics, strain 2T18T represents a novel species of the genus Chitinibacter, for which the name Chitinibacter bivalviorum sp. nov. is proposed. The type strain is 2T18T (=KCTC 72821T=CCUG 74764T).
Assuntos
Anodonta/microbiologia , Betaproteobacteria/classificação , Filogenia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , Betaproteobacteria/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Água Doce , Trato Gastrointestinal/microbiologia , Hibridização de Ácido Nucleico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/químicaRESUMO
A polyphasic taxonomic approach was used to characterize a novel bacterium, designated as strain HDW20T, isolated from the intestine of the dark diving beetle Hydrophilus acuminatus. The isolate was Gram-stain-positive, facultatively anaerobic, non-motile, coccus-shaped, and formed pale orange colonies. Phylogenetic analysis based on 16S rRNA gene sequences and genome sequences showed that the isolate belonged to the genus Tessaracoccus in the phylum Actinobacteria and was closely related to T. flavescens SST-39T, T. defluvii JCM 17540T, and T. aquimaris NSG39T, with the highest 16S rRNA gene sequence similarity of 98.5â% and a highest average nucleotide identity (ANI) value of 80.6â%. The major cellular fatty acids were C18â:â1 ω9c and anteiso-C15â:â0. The main respiratory quinone was MK-9 (H4). The major polar lipid components were phosphatidylglycerol and diphosphatidylglycerol. The genomic DNA G+C content was 69.0â%. The isolate contains ÊÊ-diaminopimelic acid, Ê-alanine, and Ê-lysine as amino acid components, and ribose, glucose, and galactose as sugar components of the cell wall peptidoglycan. The results of phylogenetic, phenotypic, chemotaxonomic, and genotypic analyses suggested that strain HDW20T represents a novel species within the genus Tessaracoccus. We propose the name Tessaracoccus coleopterorum sp. nov. The type strain is HDW20T (=KACC 21348T=KCTC 49324T=JCM 33674T).
Assuntos
Besouros/microbiologia , Intestinos/microbiologia , Filogenia , Propionibacteriaceae/classificação , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Graxos/química , Peptidoglicano/química , Fosfolipídeos/química , Pigmentação , Propionibacteriaceae/isolamento & purificação , RNA Ribossômico 16S/genética , República da Coreia , Rios , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMO
A novel Gram-stain-positive, non-motile, non-spore-forming, coccobacillus-shaped, strictly aerobic bacterium, designated strain H23T48T, was isolated from the faecal sample of an oriental stork collected from the Seoul Grand Park Zoo in Seoul, Republic of Korea. Optimal growth of strain H23T48T was observed at 30-37 °C, pH 8 and with 3â% (w/v) NaCl. 16S rRNA gene sequence-based phylogenetic analysis revealed that strain H23T48T was closely related to the genus Flaviflexus, with 97.0 and 96.7â% sequence similarities to Flaviflexus salsibiostraticola EBR4-1-2T and Flaviflexus huanghaiensis H5T, respectively. Strain H23T48T possessed MK-9(H4) as the major menaquinone and C16â:â0 (42.4â%), C18â:â1 ω9c (31.3â%) and C14â:â0 (17.7â%) as the major cellular fatty acids. The polar lipids included phosphatidylglycerol, two unidentified lipids, six unidentified phospholipids and two unidentified glycophospholipids. The amino acid composition of the cell-wall peptidoglycan was l-alanine, l-lysine, d-glutamic acid, l-aspartic acid and glycine. The genomic G+C content of strain H23T48T is 59.5 mol% and the average nucleotide identity value between H23T48T and F. salsibiostraticola KCT C33148T (=EBR4-1-2T) is 75.5â%. Based on the obtained data, strain H23T48T represents a novel species of the genus Flaviflexus, for which the name Flaviflexus ciconiae sp. nov. is proposed. The type strain is H23T48T (=KCTC 49253T=JCM 33282T).
Assuntos
Actinomycetaceae/classificação , Aves/microbiologia , Filogenia , Actinomycetaceae/isolamento & purificação , Animais , Animais de Zoológico/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Fezes/microbiologia , Peptidoglicano/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMO
A novel, Gram-stain-positive, non-spore-forming, facultatively anaerobic bacterium, designated strain H21T32T, was isolated from the faeces of an Oriental stork, Ciconia boyciana. Cells formed cocci grouped in pairs, tetrads or conglomerates, and colonies on solid medium were pale yellow. Strain H21T32T belonged to the genus Jeotgalibaca, family Carnobacteriaceae, order Lactobacillales and class Bacilli. The 16S rRNA gene sequences of the strain showed 97.06-97.34, 96.17-96.31 and 95.93-96.07â% similarity to the type strains of Jeotgalibaca arthritidis, J. porci and J. dankookensis, respectively. The strain grew at 10-37 °C (optimum temperature: 30 °C), with 0-7â% (w/v) NaCl (optimum salinity: 0.5â%) and at pH 7-9 (optimum pH: 8). The main cellular fatty acids were C16â:â1 ω9c, C18â:â1 ω9c and C16â:â0. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. Respiratory quinones were not detected. Sugar components of the peptidoglycan were rhamnose, ribose and glucose. Amino acid components of the cell wall were l-alanine, d-glucose, l-lysine, glycine and aspartic acid. The DNA G+C content of the strain was 37.1 mol%. Average nucleotide identity between strain H21T32T and J. arthritidis CECT 9157T was 77.02 %, confirming that strain H21T32T represents a novel species of the genus Jeotgalibaca, for which the name Jeotgalibaca ciconiae sp. nov. is proposed. The type strain is H21T32T (=KCTC 33991T=JCM 33222T).
Assuntos
Aves/microbiologia , Carnobacteriaceae/classificação , Fezes/microbiologia , Filogenia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , Carnobacteriaceae/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Peptidoglicano/química , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNARESUMO
A novel Gram-stain-negative, aerobic, rod-shaped, reddish-orange-coloured, gliding bacterial strain, designated L12M1T, was isolated from the gut of the Korean scallop, Patinopecten yessoensis. Phylogenetic analysis based on 16S rRNA gene sequence revealed that strain L12M1T formed a monophyletic clade with the strains in the genus Flammeovirga and showed highest 16S rRNA gene sequence similarity to Flammeovirga kamogawensis YS10T (98.66â%). The major cellular fatty acids of strain L12M1T were iso-C15â:â0 and C20â:â4ω6,9,12,15c. The predominant isoprenoid quinone was MK-7. The major polyamines were spermidine, cadaverine and the minor polyamine was putrescine. The DNA G+C content was 32.1 mol%. The phylogenetic, phenotypic, biochemical, chemotaxonomic and genotypic results indicated that strain L12M1T represents a novel species of the genus Flammeovirga, for which the name Flammeovirga pectinis sp. nov. is proposed. The type strain is L12M1T (=KCTC 62750T=JCM 33169T).
Assuntos
Bacteroidetes/classificação , Pectinidae/microbiologia , Filogenia , Animais , Técnicas de Tipagem Bacteriana , Bacteroidetes/isolamento & purificação , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Pigmentação , Poliaminas/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMO
A novel Gram-negative, obligately aerobic, rod-shaped and non-motile bacterium, designated strain K13M18T, was isolated from the intestinal tract of a Korean indigenous fish, oily bitterling (Acheilognathus koreensis). Strain K13M18T formed creamy-pink colonies on a marine agar plate. Results of phylogenetic analysis based on the 16S rRNA gene sequence similarity indicated that strain K13M18T was most closely related to Tabrizicola sediminis DRYC-M-16T, sharing 97.62 % similarity with that strain. Strain K13M18T belonged to the genus Tabrizicola, which formed a cluster with Tabrizicola aquatica RCRI19T, Tabrizicola fusiformis SY72T, Tabrizicola sediminis DRYC-M-16T and Tabrizicola alkalilacus DJCT in a phylogenetic tree based on the 16S rRNA gene sequences. Strain K13M18T grown optimally in 0 % (w/v) NaCl, at pH 7 and 30 °C, in a marine broth medium. The predominant cellular fatty acids were C18 : 1 ω7c and C18 : 1 ω6c. The major respiratory isoprenoid quinone was ubiquinone Q-10. Polar lipids of strain K13M18T contained phosphatidylcholine, phosphatidylglycerol, phosphatidylinositol, diphosphatidylglycerol, six unidentified aminophospholipids, one unidentified aminolipid and an unidentified lipid. Based on genome sequencing, the DNA G+C content of strain K13M18T was 64.08 mol %, with an average nucleotide identity value, calculated by a comparative genomic analysis of strains K13M18T and T. sediminis DRYC-M-16T, of 74.82 %. Based on the phylogenetic, genotypic, and phenotypic information, strain K13M18T is proposed to be a novel species of the genus Tabrizicola. The type strain is K13M18T (=KCTC 62659T=JCM 33230T).
Assuntos
Peixes/microbiologia , Trato Gastrointestinal/microbiologia , Filogenia , Rhodobacteraceae/classificação , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Água Doce , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , República da Coreia , Rhodobacteraceae/isolamento & purificação , Análise de Sequência de DNA , Ubiquinona/análogos & derivados , Ubiquinona/químicaRESUMO
A novel Gram-stain-negative, coccus-shaped, aerobic and motile bacterial strain, designated S12M18T, was isolated from the gut of the Korean turban shell, Turbo cornutus. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain S12M18T belonged to the genus Pseudorhodobacter and had the highest 16S rRNA gene sequence similarity twith Pseudorhodobacter aquimaris HDW-19T (98.63â%). The phylogenomic tree congruently verified that strain S12M18T occupies a taxonomic position within the genus Pseudorhodobacter. The OrthoANIu value between strain S12M18T and P. aquimaris HDW-19T was 87.22â%. The major cellular fatty acid of strain S12M18T was summed feature 8 (C18â:â1 ω7c or C18â:â1 ω6c). The major components of the polar lipids were phosphatidylcholine, phosphatidylglycerol and phosphatidylethanolamine. The predominant isoprenoid quinone was Q-10. The DNA Gï¼C content was 57.8 mol%. The polyphasic analyses indicated that strain S12M18T represents a novel species of the genus Pseudorhodobacter, for which the name Pseudorhodobacter turbinis sp. nov. is proposed. The type strain is S12M18T (=KCTC 62742T=JCM 33168T).
RESUMO
A novel Gram-stain-negative, non-spore-forming, obligate aerobic, motile, rod-shaped, and flagellated bacterium, designated S11R28T, was isolated from the intestinal tract of a Korean shiner, Coreoleuciscus splendidus. Based on 16S rRNA gene sequences, strain S11R28T was identified as member of the genus Undibacterium in class Betaproteobacteria, and was closely related to Undibacterium parvum DSM 23061T (98.49â%). The isolate grew at 4-25 °C, pH 6-9, with 0â% (w/v) NaCl, and grew optimally at 20 °C, pH 8, in the absence of NaCl. The main cellular fatty acids were C16â:â0 and summed features 3 (C16â:â1ω7c and/or C16â:â1ω6c). The strain possessed diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine as predominant polar lipids, and ubiquinone Q-8 as a respiratory quinone. The polyamine profile composed of 2-hydroxyputrescine, spermidine, putrescine, and benzoic acid. A genomic DNA G+C content was 51.4 mol%. The average nucleotide identity between strains S11R28T and U. parvum DSM 23061T was 78.66â%. Thus, Undibacterium piscinae can be considered a novel species within the genus Undibacterium with the type strain S11R28T (=KCTC 62668T=JCM 33224T).
Assuntos
Cyprinidae/microbiologia , Intestinos/microbiologia , Oxalobacteraceae/classificação , Filogenia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Oxalobacteraceae/isolamento & purificação , Fosfolipídeos/química , Poliaminas/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/químicaRESUMO
A novel Gram-stain-negative, facultatively anaerobic, non-motile, non-violet-pigmented, rod-shaped bacterium, designated strain H11R3T, was isolated from the feces of Oriental stork, Ciconia boyciana, collected from Seoul Grand Park Zoo, Republic of Korea. Phylogenetic analysis of the 16S rRNA gene sequence revealed that H11R3T formed a monophyletic clade with Iodobacter fluviatilisDSM 3764T, Iodobacter arcticusDSM 100243T, and Iodobacter limnosediminisDSM 103822T, with sequence similarities of 98.8, 98.6 and 98.4â%, respectively. H11R3T grew optimally at 15 °C, pH 8, with 0.5â% (w/v) NaCl. The predominant isoprenoid quinone was ubiquinone-8 (Q-8), and polar lipids included phosphatidylethanolamine, three unidentified lipids, four unidentified phospholipids, and two unidentified aminophospholipids. The major fatty acids were summed feature 3 and C16â:â0, and the DNA G+C content of the genome is 48.0 mol%. The average nucleotide identity (ANI) value between strains H11R3T and I. fluviatilis NCTC 11159T (=DSM 3764T) is 83.7â%. On the basis of phenotypic, genotypic, phylogenetic and chemotaxonomic characteristics, strain H11R3T represents a novel species of the genus Iodobacter for which the name Iodobacterciconiae sp. nov. is proposed. The type strain is H11R3T (=KCTC 62666T=JCM 33283T).
Assuntos
Betaproteobacteria/classificação , Aves/microbiologia , Filogenia , Animais , Animais de Zoológico/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , Betaproteobacteria/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Fezes/microbiologia , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/químicaRESUMO
Matrix metalloproteinases (MMPs) play important roles in normal brain development and synaptic plasticity, although aberrant expression of MMPs leads to brain damage, including blood-brain barrier disruption, inflammation, demyelination, and neuronal cell death. In this article, we report that MMP-8 is upregulated in LPS-stimulated BV2 microglial cells and primary cultured microglia, and treatment of MMP-8 inhibitor (M8I) or MMP-8 short hairpin RNA suppresses proinflammatory molecules, particularly TNF-α secretion. Subsequent experiments showed that MMP-8 exhibits TNF-α-converting enzyme (TACE) activity by cleaving the prodomain of TNF-α (A(74)/Q(75), A(76)/V(77) residues) and, furthermore, that M8I inhibits TACE activity more efficiently than TAPI-0, a general TACE inhibitor. Biochemical analysis of the underlying anti-inflammatory mechanisms of M8I revealed that it inhibits MAPK phosphorylation, NF-κB/AP-1 activity, and reactive oxygen species production. Further support for the proinflammatory role of microglial MMP-8 was obtained from an in vivo animal model of neuroinflammatory disorder. MMP-8 is upregulated in septic conditions, particularly in microglia. Administration of M8I or MMP-8 short hairpin RNA significantly inhibits microglial activation and expression/secretion of TNF-α in brain tissue, serum, and cerebrospinal fluid of LPS-induced septic mice. These results demonstrate that MMP-8 critically mediates microglial activation by modulating TNF-α activity, which may explain neuroinflammation in septic mouse brain.
Assuntos
Encefalopatias/imunologia , Encéfalo/imunologia , Sistema de Sinalização das MAP Quinases/imunologia , Metaloproteinase 8 da Matriz/imunologia , Proteínas do Tecido Nervoso/imunologia , Fator de Necrose Tumoral alfa/imunologia , Proteínas ADAM/imunologia , Proteína ADAM17 , Animais , Encéfalo/patologia , Encefalopatias/induzido quimicamente , Encefalopatias/patologia , MAP Quinases Reguladas por Sinal Extracelular/imunologia , Inflamação/induzido quimicamente , Inflamação/imunologia , Inflamação/patologia , Lipopolissacarídeos/toxicidade , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , Inibidores de Metaloproteinases de Matriz/farmacologia , Camundongos , Camundongos Endogâmicos ICR , Microglia/imunologia , Microglia/patologia , NF-kappa B/imunologia , Fosforilação/efeitos dos fármacos , Fosforilação/imunologia , Sepse/induzido quimicamente , Sepse/imunologia , Sepse/patologia , Fator de Transcrição AP-1/imunologiaRESUMO
The first and second preferred substrates of recombinant Escherichia coli cells expressing 10R-dioxygenase (PpoC) from Aspergillus nidulans and the purified enzyme were linoleic acid and α-linolenic acid, respectively. PpoC in cells showed higher thermal and reaction stabilities compared to purified PpoC. Thus, 10R-hydroxy unsaturated fatty acids were produced from linoleic acid, α-linolenic acid, and hempseed oil hydrolyzate containing linoleic acid and α-linolenic acid as substrates by whole recombinant cells expressing PpoC. The optimal reaction conditions for the production of 10R-hydroxy-8E,12Z-octadecadienoic acid (10R-HODE) were pH 8.0, 30 °C, 250 rpm, 5 % (v/v) dimethyl sulfoxide, 5 g l(-1) linoleic acid, and 60 g l(-1) cells in 100-ml baffled flask. Under these conditions, whole recombinant cells expressing PpoC produced 2.7 g l(-1) 10R-HODE from 5 g l(-1) linoleic acid for 40 min, with a conversion yield of 54 % (w/w) and a productivity of 4.0 g l(-1) h(-1); produced 2.2 g l(-1) 10R-hydroxy-8E,12Z,15Z-octadecatrienoic acid (10R-HOTrE) from 3 g l(-1) α-linolenic acid for 30 min, with a conversion yield of 72 % (w/w) and a productivity of 4.3 g l(-1) h(-1); and produced 1.8 g l(-1) 10R-HODE and 0.5 g l(-1) 10R-HOTrE from 5 g l(-1) hempseed oil hydrolyzate containing 2.5 g l(-1) linoleic acid and 1.0 g l(-1) α-linolenic acid for 30 min, with a conversion yield of 74 and 51 % (w/w), respectively, and a productivity of 3.6 and 1.0 g l(-1) h(-1), respectively. To the best of our knowledge, this is the first report on the biotechnological production of 10R-hydroxy unsaturated fatty acids.
Assuntos
Aspergillus nidulans/enzimologia , Dioxigenases/metabolismo , Escherichia coli/metabolismo , Ácidos Graxos Insaturados/metabolismo , Óleos de Plantas/metabolismo , Proteínas Recombinantes/metabolismo , Aspergillus nidulans/genética , Biotransformação , Dioxigenases/genética , Escherichia coli/genética , Concentração de Íons de Hidrogênio , Ácido Linoleico/metabolismo , Proteínas Recombinantes/genética , TemperaturaRESUMO
Initial and recurrent stroke produces central nervous system (CNS) damage, involving neuroinflammation. Receptor-mediated S1P signaling can influence neuroinflammation and has been implicated in cerebral ischemia through effects on the immune system. However, S1P-mediated events also occur within the brain itself where its roles during stroke have been less well studied. Here we investigated the involvement of S1P signaling in initial and recurrent stroke by using a transient middle cerebral artery occlusion/reperfusion (M/R) model combined with analyses of S1P signaling. Gene expression for S1P receptors and involved enzymes was altered during M/R, supporting changes in S1P signaling. Direct S1P microinjection into the normal CNS induced neuroglial activation, implicating S1P-initiated neuroinflammatory responses that resembled CNS changes seen during initial M/R challenge. Moreover, S1P microinjection combined with M/R potentiated brain damage, approximating a model for recurrent stroke dependent on S1P and suggesting that reduction in S1P signaling could ameliorate stroke damage. Delivery of FTY720 that removes S1P signaling with chronic exposure reduced damage in both initial and S1P-potentiated M/R-challenged brain, while reducing stroke markers like TNF-α. These results implicate direct S1P CNS signaling in the etiology of initial and recurrent stroke that can be therapeutically accessed by S1P modulators acting within the brain.
Assuntos
Isquemia Encefálica/prevenção & controle , Lisofosfolipídeos/fisiologia , Receptores de Lisoesfingolipídeo/antagonistas & inibidores , Esfingosina/análogos & derivados , Acidente Vascular Cerebral/prevenção & controle , Animais , Barreira Hematoencefálica , Isquemia Encefálica/etiologia , Cloridrato de Fingolimode/farmacologia , Lisofosfolipídeos/toxicidade , Masculino , Camundongos , Camundongos Endogâmicos ICR , Microinjeções , Neuroglia/efeitos dos fármacos , Neuroglia/fisiologia , Fosfotransferases (Aceptor do Grupo Álcool)/fisiologia , Transdução de Sinais , Esfingosina/fisiologia , Esfingosina/toxicidade , Acidente Vascular Cerebral/etiologia , Fator de Necrose Tumoral alfa/análiseRESUMO
Astrocytes are the most abundant cell-type of the human brain and play a variety of roles in brain homeostasis and synaptic maturation, under normal conditions. However, astrocytes undergo dramatic pathological changes in response to brain injury, such as reactive gliosis and glial scar formation. Although abnormal hypertrophy and massive proliferation of astrocytes are obvious, the molecular identity and cues that dictate the structural changes in reactive astrocytes remain unclear. This study proposes that fibroblast growth factor (FGF) signaling is responsible for making astrocyte morphology more complex and hypertrophic in response to an inflammatory stimulus such as lipopolysaccharide. Primary astrocytes isolated from perinatal brains developed more branches in the presence of FGF8 or lesser branches in the presence of FGF2. Introduction of the constitutively active form of the FGF receptor 3 (caFGFR3) into the brain increases the structural complexity, with greater glial fibrillary acidic protein level in astrocytes, while overexpression of a dominant-negative form of FGFR3 (dnFGFR3) reduces it. Treatment of FGF8 facilitated the wound-healing process of primary astrocytes in vitro by changing their morphology, indicating that the FGF signal may control the responsiveness of astrocytes in injury conditions. Finally, the blockade of FGF signaling by introducing dnFGFR3 at the site of reactive gliosis reduces astrocyte branch formation and minimizes hypertrophic responses during reactive gliosis. Taken together, these results indicate that FGF8-FGFR3 signaling controls structural changes in astrocytes during reactive gliosis, under pathogenic conditions.
Assuntos
Astrócitos/patologia , Astrócitos/fisiologia , Encéfalo/patologia , Encéfalo/fisiopatologia , Fatores de Crescimento de Fibroblastos/metabolismo , Animais , Células Cultivadas , Fator 2 de Crescimento de Fibroblastos/metabolismo , Fator 8 de Crescimento de Fibroblasto/metabolismo , Proteína Glial Fibrilar Ácida/metabolismo , Gliose/patologia , Gliose/fisiopatologia , Lipopolissacarídeos , Masculino , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICR , Ratos , Receptor Tipo 3 de Fator de Crescimento de Fibroblastos/metabolismo , Transdução de Sinais , Cicatrização/fisiologiaRESUMO
Background: Periprosthetic osteolysis is a prevalent complication following total ankle arthroplasty (TAA), implicating various cytokines in osteoclastogenesis as pivotal in this process. This study aimed to evaluate the relationship between osteolysis and the concentrations of osteoclastogenesis-related cytokines in synovial fluid and investigate its clinical value following TAA. Methods: Synovial fluid samples from 23 ankles that underwent revision surgery for osteolysis following TAA were analyzed as the osteolysis group. As a control group, we included synovial fluid samples obtained from 23 ankles during primary TAA for osteoarthritis. The receptor activator of nuclear factor-κB ligand (RANKL)/osteoprotegerin (OPG) ratio in these samples was quantified using sandwich enzyme-linked immunosorbent assay techniques, and a bead-based multiplex immunoassay facilitated the detection of specific osteoclastogenesis-related cytokines. Results: RANKL levels averaged 487.9 pg/mL in 14 of 23 patients in the osteolysis group, with no detection in the control group's synovial fluid. Conversely, a significant reduction in OPG levels was observed in the osteolysis group (p = 0.002), resulting in a markedly higher mean RANKL/OPG ratio (0.23) relative to controls (p = 0.020). Moreover, the osteolysis group had increased concentrations of various osteoclastogenesis-related cytokines (tumor necrosis factor-α, interleukin [IL]-1ß, IL-6, IL-8, IP-10, and monocyte chemotactic protein-1) in the synovial fluid relative to the control group. Conclusions: Our results demonstrated that periprosthetic osteolysis was associated with osteoclastogenesis activation through an elevated RANKL/OPG ratio following TAA. We assume that RANKL and other osteoclastogenesis-related cytokines in the synovial fluid have clinical value as a potential marker for the development and progression of osteolysis following TAA.
Assuntos
Artroplastia de Substituição do Tornozelo , Biomarcadores , Osteólise , Osteoprotegerina , Ligante RANK , Líquido Sinovial , Humanos , Líquido Sinovial/metabolismo , Líquido Sinovial/química , Osteólise/metabolismo , Osteólise/etiologia , Masculino , Feminino , Ligante RANK/metabolismo , Idoso , Pessoa de Meia-Idade , Artroplastia de Substituição do Tornozelo/efeitos adversos , Osteoprotegerina/metabolismo , Osteoprotegerina/análise , Biomarcadores/metabolismo , Biomarcadores/análise , Idoso de 80 Anos ou mais , Citocinas/metabolismo , Citocinas/análise , ReoperaçãoRESUMO
The chemical study of the leaves and twigs of Viscum album led to the isolation of five new lignan glycosides, namely, ligalbumosides A-E (2-6) and one known lignan glycoside, alangilignoside C (1). The structures of five new lignan glycosides were determined to be (7R,8S,8'S)-4,9,4'-trihydroxy-3,5,3',5'-tetramethoxy-7,9'-epoxylignan 9-O-ß-D-glucopyranoside (2), (7S,8S,7'S,8'R)-4,9,4'-trihydroxy-3,5,3',5',7'-pentamethoxy-7,9'-epoxylignan 9-O-ß-D-glucopyranoside (3), (7R,8R,7'S,8'S)-4,9,4'-trihydroxy-3,5,3',5',7'-pentamethoxy-7,9'-epoxylignan 9-O-ß-D-glucopyranoside (4), (7S,8R,7'S,8'R)-4,9,4'-trihydroxy-3,5,3',5',7'-pentamethoxy-7,9'-epoxylignan 9-O-ß-D-glucopyranoside (5), and (7R,8S,7'R,8'S)-4,9,4',7'-tetrahydroxy-3,5,3',5'-tetramethoxy-7,9'-epoxylignan 9-O-ß-D-glucopyranoside (6) using 1D-, 2D-NMR, and CD spectra, chemical methods, as well as comparing the results with those reported in the literature.
Assuntos
Glicosídeos/química , Viscum album/química , Dicroísmo Circular , Ligantes , Espectroscopia de Ressonância Magnética/normas , Conformação Molecular , Padrões de Referência , EstereoisomerismoRESUMO
BACKGROUND: Osteolysis is one of the most common problems that occurs after total hip and knee arthroplasty and has recently become a significant problem after total ankle arthroplasty (TAA). In this study, we investigated the role of LIM homeobox transcription factor 1-ß (Lmx1b) in osteoclast differentiation. By evaluating the expression profiles associated with osteolysis following TAA treatment, Lmx1b was found to be differentially expressed in patients with osteolysis after TAA. METHODS: To identify the important genes associated with osteolysis after TAA, RNA sequencing was performed by analyzing 8 patient samples: 5 primary TAA samples (control group) and 3 TAA samples revised for flexion instability (osteolysis group). By analyzing the differentially expressed genes and gene ontologies, Lmx1b expression was found to be upregulated in the osteolysis group compared to that in the control group. Focusing on the role of Lmx1b in bone cells, Lmx1b was overexpressed by a retrovirus in osteoclast precursor cells. The cultured cells were stained with tartrate-resistant acid phosphatase, and the expression of osteoclast-related genes was analyzed using real-time polymerase chain reaction. RESULTS: Lmx1b overexpression in osteoclast precursors suppresses osteoclast formation and resorptive activity. The expression of osteoclast marker genes was significantly reduced during osteoclast differentiation by Lmx1b overexpression. Furthermore, Lmx1b is associated with nuclear factor of activated T cells 1 (NFATc1) and inhibited NFATc1 translocation into the nucleus. CONCLUSIONS: These results provide novel insights into the anti-bone resorptive effect of Lmx1b on osteolysis after TAA and may lead to the development of effective preventative and therapeutic strategies for peri-implant osteolysis.