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1.
Plants (Basel) ; 12(18)2023 Sep 15.
Article in English | MEDLINE | ID: mdl-37765437

ABSTRACT

The present study provides new insights into the growth of the brown algal cell wall by showing that cell wall polysaccharides play an important role in the process of growth, considering the physicochemical characteristic of young and old Cladosiphon okamuranus. To determine its structural variation in detail, the cell wall was sequentially fractionated into five fractions: hot water (HW), ammonium oxalate, hemicellulose-I (HC-I), HC-II, and cellulose, and analyzed physicochemically. Results showed that almost 80% of the total recovery cell wall from both young and old thalli was HW, and HC-I contained mainly fucoidan composed of Fucose, Glucuronic acid, and sulfate in molar ratios of 1.0:0.3:0.6~0.7 and 1.0:0.3:0.2~0.3, respectively. Fucoidan in HW was a highly sulfated matrix polysaccharide abundance in young thalli, while fucoidan in HC-I was rich in old thalli and functions as hemicellulose in land plants, crosslinking with cellulose and strengthening the cell wall. We found that HW and HC-I were particularly involved in the growth and strength of old thalli appeared to be due to the deposition of HC-I and the reduction in water content during the growth process.

2.
Carbohydr Res ; 523: 108722, 2023 Jan.
Article in English | MEDLINE | ID: mdl-36459703

ABSTRACT

Brown algae contain a polysaccharide-rich cell wall, mainly composed of alginate and fucoidan which have been extensively studied for their individual structure and bioactivities. Particularly, the cell wall of Cladosiphon okamuranus is rich in fucoidan rather than alginate. However, little is known about its arrangement or interlinking with other polysaccharides such as cellulose in the cell wall. To determine its structure in detail, the cell wall was sequentially fractionated into five fractions: hot water (HW), ammonium oxalate, hemicellulose-I (HC-I), HC-II, and cellulose. Almost 80% of the total cell wall recovered from alcohol insoluble residue in C. okamuranus consisted of HW and HC-I, which mainly contained fucoidan composed of fucose, glucuronic acid, and sulfate in molar ratios of 1.0:0.3:0.9 and 1.0:0.2:0.3, respectively. Methylation analysis revealed that fucoidan in HW and HC-I structurally differed in terms of content of sulfate, and sugar residue which was 1,4-linked xylose and 1,4-linked fucose. Small angle X-ray scattering measurements also showed distinct conformational differences between HW and HC-I. These structural heterogeneities of fucoidan may be related to their localization, and fucoidan in HC-I may be involved in reinforcing cell wall structure by cross-linking to cellulose.


Subject(s)
Fucose , Phaeophyceae , Phaeophyceae/chemistry , Polysaccharides/chemistry , Cellulose , Alginates , Cell Wall , Sulfates
3.
Microbiome ; 10(1): 220, 2022 Dec 12.
Article in English | MEDLINE | ID: mdl-36503599

ABSTRACT

BACKGROUND: Endozoicomonas bacteria symbiosis with various marine organisms is hypothesized as a potential indicator of health in corals. Although many amplicon analyses using 16S rRNA gene have suggested the diversity of Endozoicomonas species, genome analysis has been limited due to contamination of host-derived sequences and difficulties in culture and metagenomic analysis. Therefore, the evolutionary and functional potential of individual Endozoicomonas species symbiotic with the same coral species remains unresolved. RESULTS: In this study, we applied a novel single-cell genomics technique using droplet microfluidics to obtain single-cell amplified genomes (SAGs) for uncultured coral-associated Endozoicomonas spp. We obtained seven novel Endozoicomonas genomes and quantitative bacterial composition from Acropora tenuis corals at four sites in Japan. Our quantitative 16S rRNA gene and comparative genomic analysis revealed that these Endozoicomonas spp. belong to different lineages (Clade A and Clade B), with widely varying abundance among individual corals. Furthermore, each Endozoicomonas species possessed various eukaryotic-like genes in clade-specific genes. It was suggested that these eukaryotic-like genes might have a potential ability of different functions in each clade, such as infection of the host coral or suppression of host immune pathways. These Endozoicomonas species may have adopted different host adaptation strategies despite living symbiotically on the same coral. CONCLUSIONS: This study suggests that coral-associated Endozoicomonas spp. on the same species of coral have different evolutional strategies and functional potentials in each species and emphasizes the need to analyze the genome of each uncultured strain in future coral-Endozoicomonas relationships studies. Video Abstract.


Subject(s)
Anthozoa , Gammaproteobacteria , Animals , Anthozoa/microbiology , RNA, Ribosomal, 16S/genetics , Host Adaptation , Gammaproteobacteria/genetics , Symbiosis , Bacteria , Genomics , Coral Reefs
5.
Mar Biotechnol (NY) ; 24(5): 871-881, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35997836

ABSTRACT

Coral symbionts are important members of the coral holobiont, and coral bacterial flora are essential in host health maintenance and coral conservation. Coral symbionts are affected by various environmental factors, such as seawater temperature, pH, and salinity. Although physicochemical and chemical factors have been highlighted as possible causes of these effects, the effects of water flow and the co-culture of different species corals have not been elucidated. In this study, we designed an artificial rearing environment to examine the impact of environmental and biological factors on Acropora tenuis, one of the major coral species in Okinawa, and Montipora digitata, during their co-culture. We intervened with the water flow to reveal that the movement of the rearing environment alters the bacterial flora of A. tenuis. During the rearing under captive environment, the alpha diversity of the coral microbiota increased, suggesting the establishment of rare bacteria from the ocean. No differences in the bacterial composition between the control and water flow groups were observed under the rearing conditions. However, the structure of the bacterial flora was significantly different in the co-culture group. Comparison of bacterial community succession strongly suggested that the differences observed were due to the suppressed transmission of bacteria from the ocean in the co-culture group. These results enhance our understanding of interactions between corals and shed light on the importance of regional differences and bacterial composition of coral flora.


Subject(s)
Anthozoa , Animals , Anthozoa/microbiology , Bacteria/genetics , Biological Factors , Coculture Techniques , Coral Reefs , RNA, Ribosomal, 16S/genetics , Water
6.
Microbiol Resour Announc ; 11(2): e0115021, 2022 Feb 17.
Article in English | MEDLINE | ID: mdl-35175121

ABSTRACT

The bacterium "Candidatus Hydrogenosomobacter endosymbioticus" is an intracellular symbiont of anaerobic scuticociliate GW7, which is associated with hydrogenosome together with methanogenic archaea. Here, we report a complete genome sequence of the symbiont consisting of 827 kbp. Knowing this sequence would contribute to the understanding of the metabolic interactions and evolution of the tripartite symbiosis.

7.
Mar Biotechnol (NY) ; 23(4): 576-589, 2021 Aug.
Article in English | MEDLINE | ID: mdl-34275003

ABSTRACT

Coral microbial flora has been attracting attention because of their potential to protect corals from environmental stresses or pathogens. Although coral-associated bacteria are considered to be acquired from seawater, little is known about the relationships between microbial composition in corals and its surrounding seawater. Here, we tested several methods to identify coral-associated bacteria in coral and its surrounding seawater to detect specific types of Ruegeria species, some of which exhibit growth inhibition activities against the coral pathogen Vibrio coralliilyticus. We first isolated coral-associated bacteria from the reef-building coral Galaxea fascicularis collected at Sesoko Island, Okinawa, Japan, via random colony picking, which showed the existence of varieties of bacteria including Ruegeria species. Using newly constructed primers for colony PCR, several Ruegeria species were successfully isolated from G. fascicularis and seawater. We further investigated the seawater microbiome in association with the distance from coral reefs. By seasonal sampling, it was suggested that the seawater microbiome is more affected by seasonality than the distance from coral reefs. These methods and results may contribute to investigating and understanding the relationships between the presence of corals and microbial diversity in seawater, in addition to the efficient isolation of specific bacterial species from coral or its surrounding seawater.


Subject(s)
Anthozoa/microbiology , Rhodobacteraceae/isolation & purification , Seawater/microbiology , Animals , Coral Reefs , DNA, Environmental/analysis , Genome, Bacterial , Japan , Microbiota , Polymerase Chain Reaction , Probiotics , Rhodobacteraceae/genetics , Seasons , Vibrio
8.
Toxins (Basel) ; 12(6)2020 06 16.
Article in English | MEDLINE | ID: mdl-32560237

ABSTRACT

Fusarium head blight (FHB) of cereals is a severe disease caused by the Fusarium graminearum species complex. It leads to the accumulation of the mycotoxin deoxynivalenol (DON) in grains and other plant tissues and causes substantial economic losses throughout the world. DON is one of the most troublesome mycotoxins because it is a virulence factor to host plants, including wheat, and exhibits toxicity to plants and animals. To control both FHB and DON accumulation, a biological control approach using DON-degrading bacteria (DDBs) is promising. Here, we performed a disease control assay using an in vitro petri dish test composed of germinated wheat seeds inoculated with F. graminearum (Fg) and DDBs. Determination of both grown leaf lengths and hyphal lesion lengths as a measure of disease severity showed that the inoculation of seeds with the DDBs Devosia sp. strain NKJ1 and Nocardioides spp. strains SS3 or SS4 were protective against the leaf growth inhibition caused by Fg. Furthermore, it was as effective against DON accumulation. The inoculation with strains SS3 or SS4 also reduced the inhibitory effect on leaves treated with 10 µg mL-1 DON solution (without Fg). These results indicate that the DDBs partially suppress the disease by degrading DON.


Subject(s)
Edible Grain/microbiology , Fusarium/metabolism , Nocardioides/metabolism , Pest Control, Biological , Plant Diseases/prevention & control , Trichothecenes/metabolism , Triticum/microbiology , Germination , Plant Diseases/microbiology , Plant Leaves/microbiology , Seeds/microbiology
9.
Appl Environ Microbiol ; 85(24)2019 12 15.
Article in English | MEDLINE | ID: mdl-31585988

ABSTRACT

A number of anaerobic ciliates, unicellular eukaryotes, intracellularly possess methanogenic archaea and bacteria as symbiotic partners. Although this tripartite relationship is of interest in terms of the fact that each participant is from a different domain, the difficulty in culture and maintenance of those host species with symbiotic partners has disturbed both ecological and functional studies so far. In this study, we obtained a stable culture of a small anaerobic scuticociliate, strain GW7. By transmission electron microscopic observation and fluorescent in situ hybridization with domain-specific probes, we demonstrate that GW7 possesses both archaeal and bacterial endosymbionts in its cytoplasm. These endosymbionts are in dependently associated with hydrogenosomes, which are organelle producing hydrogen and ATP under anaerobic conditions. Clone library analyses targeting prokaryotic 16S rRNA genes, fluorescent in situ hybridization with endosymbiont-specific probes, and molecular phylogenetic analyses revealed the phylogenetic affiliations and intracellular localizations of these endosymbionts. The endosymbiotic archaeon is a methanogen belonging to the genus Methanoregula (order Methanomicrobiales); a member of this genus has previously been described as the endosymbiont of an anaerobic ciliate from the genus Metopus (class Armophorea), which is only distantly related to strain GW7 (class Oligohymenophorea). The endosymbiotic bacterium belongs to the family Holosporaceae of the class Alphaproteobacteria, which also comprises several endosymbionts of various aerobic ciliates. For this endosymbiotic bacterium, we propose a novel candidate genus and species, "Candidatus Hydrogenosomobacter endosymbioticus."IMPORTANCE Tripartite symbioses between anaerobic ciliated protists and their intracellular archaeal and bacterial symbionts are not uncommon, but most reports have been based mainly on microscopic observations. Deeper insights into the function, ecology, and evolution of these fascinating symbioses involving partners from all three domains of life have been hampered by the difficulties of culturing anaerobic ciliates in the laboratory and the frequent loss of their prokaryotic partners during long-term cultivation. In the present study, we report the isolation of an anaerobic scuticociliate, strain GW7, which has been stably maintained in our laboratory for more than 3 years without losing either of its endosymbionts. Unexpectedly, molecular characterization of the endosymbionts revealed that the bacterial partner of GW7 is phylogenetically related to intranuclear endosymbionts of aerobic ciliates. This strain will enable future genomic, transcriptomic, and proteomic analyses of the interactions in this tripartite symbiosis and a comparison with endosymbioses in aerobic ciliates.


Subject(s)
Alphaproteobacteria/metabolism , Anaerobiosis/physiology , Ciliophora/microbiology , Euryarchaeota/metabolism , Holosporaceae/physiology , Organelles/microbiology , Symbiosis , Alphaproteobacteria/classification , Alphaproteobacteria/genetics , Alphaproteobacteria/isolation & purification , Culture Media/chemistry , Euryarchaeota/classification , Euryarchaeota/genetics , Holosporaceae/classification , Holosporaceae/genetics , In Situ Hybridization, Fluorescence , Phylogeny , RNA, Ribosomal, 16S/genetics , RNA, Ribosomal, 16S/isolation & purification , Sequence Analysis, DNA
10.
Article in English | MEDLINE | ID: mdl-30863825

ABSTRACT

Actinomycete Nocardioides sp. strain LS1, isolated from wheat leaf, is a bacterium that degrades and assimilates the mycotoxin deoxynivalenol (DON) as the carbon source. This is the first study of the genome sequence of the DON-degrading genus Nocardioides, and it facilitates the study of genes encoding the DON-degrading pathway.

11.
Sci Rep ; 9(1): 2129, 2019 02 14.
Article in English | MEDLINE | ID: mdl-30765748

ABSTRACT

Gut bacteria of phytophagous and omnivorous marine invertebrates often possess alginate lyases (ALGs), which are key enzymes for utilizing macroalgae as carbon neutral biomass. We hypothesized that the exclusive feeding of a target alga to marine invertebrates would shift the gut bacterial diversity suitable for degrading the algal components. To test this hypothesis, we reared sea hare (Dolabella auricularia) and sea snail (Batillus cornutus) for two to four weeks with exclusive feeding of a brown alga (Ecklonia cava). Pyrosequencing analysis of the gut bacterial 16S rRNA genes revealed shifts in the gut microbiota after rearing, mainly due to a decrease in the variety of bacterial members. Significant increases in six and four 16S rRNA gene phylotypes were observed in the reared sea hares and sea snails, respectively, and some of them were phylogenetically close to known alginate-degrading bacteria. Clone library analysis of PL7 family ALG genes using newly designed degenerate primer sets detected a total of 50 ALG gene phylotypes based on 90% amino acid identity. The number of ALG gene phylotypes increased in the reared sea hare but decreased in reared sea snail samples, and no phylotype was shared between them. Out of the 50 phylotypes, 15 were detected only after the feeding procedure. Thus, controlled feeding strategy may be valid and useful for the efficient screening of genes suitable for target alga fermentation.


Subject(s)
Aquatic Organisms/metabolism , Bacteria/metabolism , Gastrointestinal Microbiome , Gastropoda/metabolism , Phaeophyceae/metabolism , Polysaccharide-Lyases/metabolism , Animals , Aquatic Organisms/growth & development , Aquatic Organisms/microbiology , Bacteria/genetics , DNA, Bacterial/analysis , DNA, Bacterial/genetics , Gastropoda/growth & development , Gastropoda/microbiology , Polysaccharide-Lyases/genetics
12.
Spine J ; 19(2): e34-e40, 2019 02.
Article in English | MEDLINE | ID: mdl-28735763

ABSTRACT

BACKGROUND CONTEXT: Platelet-rich plasma (PRP) accelerates bone union in vivo in a rodent model of spinal fusion surgery. However, PRP's effect on bone union after spinal surgery remains unclear. PURPOSE: The objective of this study was to evaluate the efficacy of PRP after posterolateral lumbar fusion (PLF) surgery. STUDY DESIGN/SETTING: Single-center prospective randomized controlled clinical trial with 2-year follow-up. PATIENT SAMPLE: The patient sample included a total 62 patients (31 patients in the PRP group or 31 patients in the control group). OUTCOME MEASURES: The outcome measures included the bone fusion rate, the area of bone fusion mass, the duration of bone fusion, and the clinical score using the visual analog scale (VAS). MATERIALS AND METHODS: We randomized 62 patients who underwent one- or two-level instrumented PLF for lumbar degenerative spondylosis with instability to either the PRP (31 patients) or the control (31 patients) groups. Platelet-rich plasma-treated patients underwent surgery using an autograft bone chip (local bone), and PRP was prepared from patient blood samples immediately before surgery; patients from the control group underwent PLF without PRP treatment. We assessed platelet counts and growth factor concentrations in PRP prepared immediately before surgery. The duration of bone union, the postoperative bone fusion rate, and the area of fusion mass were assessed using plain radiography every 3 months after surgery and by computed tomography at 12 or 24 months. The duration of bone fusion and the clinical scores for low back pain, leg pain, and leg numbness before and 3, 6, 12, and 24 months after surgery were evaluated using VAS. RESULTS: Data from 50 patients with complete data were included. The bone union rate at the final follow-up was significantly higher in the PRP group (94%) than in the control group (74%) (p=.002). The area of fusion mass was significantly higher in the PRP group (572 mm2) than in the control group (367 mm2) (p=.02). The mean period necessary for union was 7.8 months in the PRP group and 9.8 months in the control group (p=.013). In the PRP, the platelet count was 7.7 times higher and the growth factor concentrations were 50 times higher than those found in plasma (p<.05). There was no significant difference in low back pain, leg pain, and leg numbness in either group at any time evaluated (p>.05). CONCLUSIONS: Patients treated with PRP showed a higher fusion rate, greater fusion mass, and more rapid bone union after spinal fusion surgery than patients not treated with PRP.


Subject(s)
Bone Transplantation/methods , Platelet-Rich Plasma , Postoperative Complications/epidemiology , Spinal Fusion/methods , Adult , Aged , Bone Transplantation/adverse effects , Female , Humans , Lumbar Vertebrae/surgery , Male , Middle Aged , Spinal Fusion/adverse effects , Transplantation, Homologous/adverse effects , Transplantation, Homologous/methods
13.
Asian Spine J ; 12(1): 112-118, 2018 Feb.
Article in English | MEDLINE | ID: mdl-29503690

ABSTRACT

STUDY DESIGN: Retrospective case series. PURPOSE: To examine the efficacy of platelet-rich plasma (PRP) for bone fusion in transforaminal lumbar interbody fusion (TLIF) using local bone grafting. OVERVIEW OF LITERATURE: Several authors have reported the efficacy of PRP for bone union in animal models. However, the use of PRP for bone fusion in TLIF surgery has not been fully explored. METHODS: Twenty patients underwent single-level TLIF surgery because of L4 spondylolisthesis. An interbody fusion cage and local bone were used in nine patients (control group) and an interbody fusion cage, local bone, and PRP were used in 11 patients (PRP group). PRP was prepared from the patients' blood samples (400 mL) immediately before surgery. The duration of bone union and postoperative bone fusion rate were assessed using plain radiography at every 3 months postoperatively and computed tomography at 12 or 24 months postoperatively, respectively. Lower back pain, leg pain, and leg numbness were evaluated using the visual analog scale preoperatively and at 3, 6, 12, and 24 months postoperatively. RESULTS: The platelet count was 8.7 times higher in PRP than in blood. The bone union rate was significantly superior in the PRP group than in the control group (91% and 77%, respectively; p=0.035), whereas the average duration of bone union was not significantly different between the groups (7.7±0.74 and 10.0±2.00 months, respectively; p=0.131). There was no significant difference in lower back pain, leg pain, and leg numbness in both groups during follow-up (p>0.05). CONCLUSIONS: Our study suggests that the use of PRP in TLIF surgery increases bone fusion rate.

14.
Asian Spine J ; 11(3): 329-336, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28670400

ABSTRACT

STUDY DESIGN: Controlled laboratory study. PURPOSE: This study aimed to evaluate the efficacy of platelet-rich plasma (PRP) stored at room temperature (RT), frozen, or after freeze-drying. OVERVIEW OF LITERATURE: PRP enriches tissue repair and regeneration, and is a novel treatment option for musculoskeletal pathologies. However, whether biological activity is preserved during PRP storage remains uncertain. METHODS: PRP was prepared from blood of 12 healthy human volunteers (200 mL/person) and stored using three methods: PRP was stored at RT with shaking, PRP was frozen and stored at -80℃, or PRP was freeze-dried and stored at RT. Platelet counts and growth factor content were examined immediately after preparation, as well as 2, 4, and 8 weeks after storage. Platelet activation rate was quantified by flow cytometry. RESULTS: Platelet counts were impossible to determine in many RT samples after 2 weeks, but they remained at constant levels in frozen and freeze-dried samples, even after 8 weeks of storage. Flow cytometry showed approximately 80% activation of the platelets regardless of storage conditions. Almost no growth factors were detected in the RT samples after 8 weeks, while low but significant expression was detected in the frozen and freeze-dried PRP. Over time, the mean relative concentrations of various growth factors decreased significantly or disappeared in the RT group. In the frozen group, levels were maintained for 4 weeks, but decreased significantly by 8 weeks (p <0.05). The freeze-dried group maintained baseline levels of growth factors for the entire 8-week duration. CONCLUSIONS: Freeze-drying enables PRP storage while maintaining bioactivity and efficacy for extended periods.

15.
J Biosci Bioeng ; 120(2): 174-80, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25656071

ABSTRACT

Bacterial xylose isomerases (XI) are promising resources for efficient biofuel production from xylose in lignocellulosic biomass. Here, we investigated xylose isomerase gene (xylA) diversity in three soil metagenomes differing in plant vegetation and geographical location, using an amplicon pyrosequencing approach and two newly-designed primer sets. A total of 158,555 reads from three metagenomic DNA replicates for each soil sample were classified into 1127 phylotypes, detected in triplicate and defined by 90% amino acid identity. The phylotype coverage was estimated to be within the range of 84.0-92.7%. The xylA gene phylotypes obtained were phylogenetically distributed across the two known xylA groups. They shared 49-100% identities with their closest-related XI sequences in GenBank. Phylotypes demonstrating <90% identity with known XIs in the database accounted for 89% of the total xylA phylotypes. The differences among xylA members and compositions within each soil sample were significantly smaller than they were between different soils based on a UniFrac distance analysis, suggesting soil-specific xylA genotypes and taxonomic compositions. The differences among xylA members and their compositions in the soil were strongly correlated with 16S rRNA variation between soil samples, also assessed by amplicon pyrosequencing. This is the first report of xylA diversity in environmental samples assessed by amplicon pyrosequencing. Our data provide information regarding xylA diversity in nature, and can be a basis for the screening of novel xylA genotypes for practical applications.


Subject(s)
Aldose-Ketose Isomerases/genetics , Genes, Bacterial/genetics , Genetic Variation/genetics , Metagenome/genetics , Metagenomics , Aldose-Ketose Isomerases/classification , Amino Acid Sequence , Base Sequence , Biofuels/supply & distribution , DNA Primers/genetics , DNA, Bacterial/genetics , Databases, Nucleic Acid , Lignin/metabolism , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Soil Microbiology , Xylose/metabolism
16.
AMB Express ; 4: 72, 2014.
Article in English | MEDLINE | ID: mdl-25401073

ABSTRACT

Two haloalkane dehalogenases, LinBUT and LinBMI, each with 296 amino acid residues, exhibit only seven amino acid residue differences between them, but LinBMI's catalytic performance towards ß-hexachlorocyclohexane (ß-HCH) is considerably higher than LinBUT's. To elucidate the molecular basis governing this difference, intermediate mutants between LinBUT and LinBMI were constructed and kinetically characterized. The activities of LinBUT-based mutants gradually increased by cumulative mutations into LinBUT, and the effects of the individual amino acid substitutions depended on combination with other mutations. These results indicated that LinBUT's ß-HCH degradation activity can be enhanced in a stepwise manner by the accumulation of point mutations.

17.
Appl Environ Microbiol ; 79(5): 1619-28, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23275503

ABSTRACT

Deoxynivalenol (DON) is a natural toxin of fungi that cause Fusarium head blight disease of wheat and other small-grain cereals. DON accumulates in infected grains and promotes the spread of the infection on wheat, posing serious problems to grain production. The elucidation of DON-catabolic genes and enzymes in DON-degrading microbes will provide new approaches to decrease DON contamination. Here, we report a cytochrome P450 system capable of catabolizing DON in Sphingomonas sp. strain KSM1, a DON-utilizing bacterium newly isolated from lake water. The P450 gene ddnA was cloned through an activity-based screening of a KSM1 genomic library. The genes of its redox partner candidates (flavin adenine dinucleotide [FAD]-dependent ferredoxin reductase and mitochondrial-type [2Fe-2S] ferredoxin) were not found adjacent to ddnA; the redox partner candidates were further cloned separately based on conserved motifs. The DON-catabolic activity was reconstituted in vitro in an electron transfer chain comprising the three enzymes and NADH, with a catalytic efficiency (k(cat)/K(m)) of 6.4 mM(-1) s(-1). The reaction product was identified as 16-hydroxy-deoxynivalenol. A bioassay using wheat seedlings revealed that the hydroxylation dramatically reduced the toxicity of DON to wheat. The enzyme system showed similar catalytic efficiencies toward nivalenol and 3-acetyl deoxynivalenol, toxins that frequently cooccur with DON. These findings identify an enzyme system that catabolizes DON, leading to reduced phytotoxicity to wheat.


Subject(s)
Bacterial Proteins/metabolism , Cytochrome P-450 Enzyme System/metabolism , Sphingomonas/enzymology , Trichothecenes/metabolism , Bacterial Proteins/genetics , Cloning, Molecular , Coenzymes/metabolism , Cytochrome P-450 Enzyme System/genetics , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Kinetics , Molecular Sequence Data , NAD/metabolism , Oxidation-Reduction , Sequence Analysis, DNA , Sphingomonas/isolation & purification , Water Microbiology
18.
Cytokine ; 61(1): 112-7, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23046618

ABSTRACT

OBJECTIVE: Triggering receptor expressed on myeloid cells-1 (TREM-1) was reported to play a key roll in amplification of production of inflammatory cytokines. TREM-1 is suggested to be a specific biomarker for sepsis for this reason, but the clinical significance of TREM-1 has not been elucidated. We investigated TREM-1 expression on the cell-surface, and plasma levels of soluble TREM-1 (sTREM-1) in patients with non-infectious systemic inflammatory response syndrome (SIRS) and sepsis admitted to the ICU. METHODS: Thirty-five patients with SIRS and 21 patients with sepsis admitted to ICU were subjected to the study. TREM-1 expressions on the surfaces of monocytes and neutrophils were measured by flow cytometry. Plasma sTREM-1 level and serum interleukin (IL)-6 level were measured. RESULTS: Septic patients had decreased TREM-1 expression, clearly on neutrophils or to a lesser extent on monocyte compared to SIRS patients on ICU admission (neutrophils p<0.001, monocyte p<0.05). TREM-1 expression on neutrophils had a significant inverse correlation with serum IL-6 level (r=-0.64, p<0.0001). Plasma sTREM-1 level in septic patients was significantly higher than that in SIRS patients (p<0.05). Plasma sTREM-1 level positively correlated with severity score and non-survivors had increased plasma sTREM-1 level compared to survivors in all SIRS/sepsis patients (p<0.05). CONCLUSIONS: Patients with sepsis had increased soluble TREM-1 and decreased TREM-1 expression on neutrophil compared to SIRS patients. sTREM-1 may be useful to evaluate disease severity and outcome of patients with SIRS or sepsis.


Subject(s)
Membrane Glycoproteins/metabolism , Monocytes/metabolism , Neutrophils/metabolism , Receptors, Immunologic/metabolism , Sepsis/metabolism , Systemic Inflammatory Response Syndrome/metabolism , Aged , Biomarkers/blood , Biomarkers/metabolism , Cytokines/biosynthesis , Female , Humans , Interleukin-6/blood , Male , Membrane Glycoproteins/blood , Membrane Proteins/biosynthesis , Membrane Proteins/metabolism , Middle Aged , Receptors, Immunologic/blood , Sepsis/blood , Systemic Inflammatory Response Syndrome/blood , Triggering Receptor Expressed on Myeloid Cells-1
19.
Masui ; 61(3): 252-8, 2012 Mar.
Article in Japanese | MEDLINE | ID: mdl-22571119

ABSTRACT

Establishing cooperative relationship is critical for departments associated with blood transfusion for massive intraoperative hemorrhage, because it is directly linked to saving lives. To achieve this aim, careful preparation and establishment of in-house rules based on the "Guidelines for Management of Critical Bleeding", depending on site-specific situations, are necessary. Blood transfusion departments should (1) share recognition with other departments in the hospital, (2) educate practitioners in blood transfusion, and (3) exchange information with blood centers. In addition, it should be recognized that regular simulations are indispensable for ensuring prompt response to emergency situations.


Subject(s)
Blood Loss, Surgical , Blood Transfusion , Patient Care Team , Female , Guidelines as Topic , Humans , Pregnancy
20.
Appl Microbiol Biotechnol ; 96(4): 1059-70, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22322873

ABSTRACT

Deoxynivalenol (DON) is a hazardous and globally prevalent mycotoxin in cereals. It commonly accumulates in the grain of wheat, barley and other small grain cereals affected by Fusarium head blight (caused by several Fusarium species). The concept of reducing DON in naturally contaminated grain of wheat or barley using a DON-degrading bacterium is promising but has not been accomplished. In this study, we isolated a novel DON-utilising actinomycete, Marmoricola sp. strain MIM116, from wheat heads through a novel isolation procedure including an in situ plant enrichment step. Strain MIM116 had background degradation activity, and the activity was enhanced twofold by the consumption of DON. Among Tween 20, Triton X-100 and Tween 80, we selected Tween 80 as a spreading agent of strain MIM116 because it promoted DON degradation and the growth of strain MIM116 in the presence of DON. The inoculation of MIM116 cell suspension plus 0.01% Tween 80 into 1,000 harvested kernels of wheat and barley resulted in a DON decrease from approximately 3 mg kg(-1) to less than 1 mg kg(-1) of dry kernels, even when cells had only basal levels of DON-degrading activity. To the best of our knowledge, this is the first report that describes (1) the isolation of a DON-degrading bacterium from wheat heads, (2) the effects of surfactants on the biodegradation of DON and (3) the decrease of DON levels in naturally contaminated wheat and barley grain using a DON-degrading bacterium.


Subject(s)
Actinomycetales/metabolism , Fusarium/physiology , Hordeum/microbiology , Mycotoxins/metabolism , Plant Diseases/microbiology , Trichothecenes/metabolism , Triticum/microbiology , Actinomycetales/classification , Actinomycetales/genetics , Actinomycetales/isolation & purification , Molecular Sequence Data , Phylogeny , Seeds/microbiology
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