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1.
Microbes Environ ; 38(4)2023.
Article in English | MEDLINE | ID: mdl-38092410

ABSTRACT

Nitrification is a key process in the biogeochemical nitrogen cycle and a major emission source of the greenhouse gas nitrous oxide (N2O). The periplasmic enzyme hydroxylamine oxidoreductase (HAO) is involved in the oxidation of hydroxylamine to nitric oxide in the second step of nitrification, producing N2O as a byproduct. Its three-dimensional structure demonstrates that slight differences in HAO active site residues have inhibitor effects. Therefore, a more detailed understanding of the diversity of HAO active site residues in soil microorganisms is important for the development of novel nitrification inhibitors using structure-guided drug design. However, this has not yet been examined. In the present study, we investigated hao gene diversity in beta-proteobacterial ammonia-oxidizing bacteria (ß-AOB) and complete ammonia-oxidizing (comammox; Nitrospira spp.) bacteria in agricultural fields using a clone library ana-lysis. A total of 1,949 hao gene sequences revealed that hao gene diversity in ß-AOB and comammox bacteria was affected by the fertilizer treatment and field type, respectively. Moreover, hao sequences showed the almost complete conservation of the six HAO active site residues in both ß-AOB and comammox bacteria. The diversity of nitrifying bacteria showed similarity between hao and amoA genes. The nxrB amplicon sequence revealed the dominance of Nitrospira cluster II in tea field soils. The present study is the first to reveal hao gene diversity in agricultural soils, which will accelerate the efficient screening of HAO inhibitors and evaluations of their suppressive effects on nitrification in agricultural soils.


Subject(s)
Archaea , Betaproteobacteria , Archaea/genetics , Soil/chemistry , Ammonia , Hydroxylamine , Catalytic Domain , Bacteria/genetics , Nitrification , Oxidation-Reduction , Hydroxylamines/pharmacology , Soil Microbiology , Phylogeny
2.
J Neurogastroenterol Motil ; 29(2): 229-237, 2023 Apr 30.
Article in English | MEDLINE | ID: mdl-37019867

ABSTRACT

Background/Aims: Symptoms of irritable bowel syndrome (IBS), a common gut-brain interaction disorder, deteriorate patients' quality of life and increase medical needs; therefore, IBS represents a significant global burden. The estimated global prevalence is approximately 10%; however, accumulated evidence shows international heterogeneity. In this study, we have described and compared the prevalence of IBS in 3 East Asian countries: Japan (Tokyo and Fukuoka), China (Beijing), and South Korea (Seoul). Methods: We conducted an internet-based cross-sectional survey of the urban population aged > 20 years in the abovementioned countries. We recruited equal numbers of age- (20s-60s) and sex-matched participants (3910 residents). IBS was diagnosed according to the Rome III criteria, and the subtypes were analyzed. Results: The overall prevalence of IBS with 95% CI was 12.6% (11.6-13.7); the prevalence was significantly different across Japan, China, and South Korea (14.9% [13.4-16.5], 5.5% [4.3-7.1], and 15.6% [13.3-18.3], respectively) (P < 0.001). Furthermore, 54.9% of patients were male. IBS-mixed was the most prevalent subtype; the prevalence of other subtypes varied. Conclusions: The overall prevalence of IBS in the 3 countries was slightly higher than the global prevalence, and it was significantly lower in China than in Japan and South Korea. IBS prevalence was the highest and lowest among individuals in their 40s and 60s, respectively. Male individuals had a higher prevalence of IBS with diarrhea. Further studies are needed to elucidate the factors associated with this regional heterogeneity.

3.
J Neuropathol Exp Neurol ; 82(1): 49-56, 2022 12 19.
Article in English | MEDLINE | ID: mdl-36383185

ABSTRACT

Intracranial aneurysms (IA) are major causes of devastating subarachnoid hemorrhages. They are characterized by a chronic inflammatory process in the intracranial arterial walls triggered and modified by hemodynamic force loading. Because IA lesion morphology is complex, the blood flow conditions loaded on endothelial cells in each portion of the lesion in situ vary greatly. We created a 3D-casted mold of the human unruptured IA lesion and cultured endothelial cells on this model; it was then perfused with culture media to model physiological flow conditions. Gene expression profiles of endothelial cells in each part of the IA lesion were then analyzed. Comprehensive gene expression profile analysis revealed similar gene expression patterns in endothelial cells from each part of the IA lesion but gene ontology analysis revealed endothelial cell malfunction within the IA lesion. Histopathological examination, electron microscopy, and immunohistochemical analysis indicated that endothelial cells within IA lesions are damaged and dysfunctional. Thus, our findings reveal endothelial cell malfunction in IA lesions and provided new insights into IA pathogenesis.


Subject(s)
Intracranial Aneurysm , Humans , Intracranial Aneurysm/pathology , Endothelial Cells/metabolism , Gene Expression Profiling , Inflammation , Transcriptome
4.
Sensors (Basel) ; 22(12)2022 Jun 07.
Article in English | MEDLINE | ID: mdl-35746105

ABSTRACT

We developed a path-planning system for radiation source identification devices using 4π gamma imaging. The estimated source location and activity were calculated by an integrated simulation model by using 4π gamma images at multiple measurement positions. Using these calculated values, a prediction model to estimate the probability of identification at the next measurement position was created by via random forest analysis. The path-planning system based on the prediction model was verified by integrated simulation and experiment for a 137Cs point source. The results showed that 137Cs point sources were identified using the few measurement positions suggested by the path-planning system.


Subject(s)
Diagnostic Imaging , Radiotherapy Planning, Computer-Assisted , Radioisotopes , Radiotherapy Dosage , Radiotherapy Planning, Computer-Assisted/methods
5.
mBio ; 13(3): e0125522, 2022 06 28.
Article in English | MEDLINE | ID: mdl-35608299

ABSTRACT

Methane-oxidizing bacteria (methanotrophs) play an ecological role in methane and nitrogen fluxes because they are capable of nitrogen fixation and methane oxidation, as indicated by genomic and cultivation-dependent studies. However, the chemical relationships between methanotrophy and diazotrophy and aerobic and anaerobic reactions, respectively, in methanotrophs remain unclear. No study has demonstrated the cooccurrence of both bioactivities in a single methanotroph bacterium in its natural environment. Here, we demonstrate that both bioactivities in type II methanotrophs occur at the single-cell level in the root tissues of paddy rice (Oryza sativa L. cv. Nipponbare). We first verified that difluoromethane, an inhibitor of methane monooxygenase, affected methane oxidation in rice roots. The results indicated that methane assimilation in the roots mostly occurred due to oxygen-dependent processes. Moreover, the results indicated that methane oxidation-dependent and methane oxidation-independent nitrogen fixation concurrently occurred in bulk root tissues. Subsequently, we performed fluorescence in situ hybridization and NanoSIMS analyses, which revealed that single cells of type II methanotrophs (involving six amplicon sequence variants) in paddy rice roots simultaneously and logarithmically fixed stable isotope gases 15N2 and 13CH4 during incubation periods of 0, 23, and 42 h, providing in vivo functional evidence of nitrogen fixation in methanotrophic cells. Furthermore, 15N enrichment in type II methanotrophs at 42 h varied among cells with an increase in 13C accumulation, suggesting that either the release of fixed nitrogen into root systems or methanotroph metabolic specialization is dependent on different microenvironmental niches in the root. IMPORTANCE Atmospheric methane concentrations have been continually increasing, causing methane to become a considerable environmental concern. Methanotrophy may be the key to regulating methane fluxes. Although research suggests that type II methanotrophs are involved in methane oxidation aerobically and nitrogen fixation anaerobically, direct evidence of simultaneous aerobic and anaerobic bioreactions of methanotrophs in situ is still lacking. In this study, a single-cell isotope analysis was performed to demonstrate these in vivo parallel functions of type II methanotrophs in the root tissues of paddy rice (Oryza sativa L. cv. Nipponbare). The results of this study indicated that methanotrophs might provide fixed nitrogen to root systems or depend on cells present in the spatially localized niche of the root tissue. Furthermore, our results suggested that single type II methanotrophic cells performed simultaneous methane oxidation and nitrogen fixation in vivo. Under natural conditions, however, nitrogen accumulation varied at the single-cell level.


Subject(s)
Oryza , In Situ Hybridization, Fluorescence , Isotopes , Methane/metabolism , Nitrogen/metabolism , Oryza/microbiology , Oxidation-Reduction , Soil Microbiology
6.
J Infect Chemother ; 28(2): 286-289, 2022 Feb.
Article in English | MEDLINE | ID: mdl-34598877

ABSTRACT

Lemierre's syndrome is a serious disease that typically causes oropharyngeal infection with internal jugular vein thrombosis, followed by distant infection focus, such as septic pulmonary embolism. The main causative organisms are anaerobic bacteria in the oral cavity, namely Fusobacterium necrophorum. We encountered an extremely rare case of Lemierre's syndrome, where double vision was found to be the first symptom. The patient's blood culture results showed the presence of F. nucleatum, which spread from the sphenoid sinus to the skull base because of chronic sinusitis; the patient presented with longus colli abscess, clivus osteomyelitis, venous thrombosis, and hematogenous infection. Antibiotic treatment with sulbactam/ampicillin was continued for 14 weeks, and no recurrence has been observed so far. Lemierre's syndrome can be complicated with atypical symptoms such as double vision if the cranial nerves are involved. It might be important to consider this disease in the differential diagnosis in the presence of cranial nerve symptoms of unknown origin with fever or inflammatory findings.


Subject(s)
Lemierre Syndrome , Venous Thrombosis , Blood Culture , Diplopia , Fusobacterium necrophorum , Humans , Jugular Veins/diagnostic imaging , Lemierre Syndrome/diagnosis , Lemierre Syndrome/drug therapy
7.
J Mater Chem B ; 9(22): 4480-4487, 2021 06 02.
Article in English | MEDLINE | ID: mdl-34031681

ABSTRACT

Biomimetic phospholipid copolymer films are known to possess antifouling properties against protein adsorption and biofilm formation. However, the interactions between bacterial cells and material surfaces are not fully understood. This work investigated the bacterial adhesion strength of phospholipid copolymer films using a shear stress-tunable microfluidic device. The copolymer, comprising 2-methacryloyloxyethyl phosphorylcholine (MPC), 3-methacryloxypropyl trimethoxysilane (MPTMSi), and 3-(methacryloyloxy) propyl-tris(trimethylsilyloxy) silane (MPTSSi), formed crosslinked films on glass substrates; the thickness of the coating film was controlled by the polymer concentration during dip-coating. Polymer films with two typical thicknesses, 20 and 40 nm (denoted as C-20 and C-40, respectively), were prepared on the bottom wall of the microfluidic device. After seeding S. aureus in the microfluidic device, several shear stresses were applied to evaluate the adhesion strength of the polymer films. S. aureus was found to have weaker adhesion strength on the C-40 surface than on the C-20 surface; numerous bacterial cells detached from the C-40 surface on application of identical shear stress. To mimic the presence of plasma protein, fibrinogen (Fg) was introduced into the device before performing the bacterial adhesion assay. The results showed that the adsorption of Fg promoted S. aureus adhesion and strong interactions under shear stress. However, the adhesion strength of S. aureus did not affect the Fg adsorption for both the C-20 and C-40 surfaces. Using the shear stress-tunable microfluidic device, we found that the adhesion of S. aureus on the thicker and softer phospholipid copolymer was weak, and the cells easily detached under high shear stress.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Adhesion , Lab-On-A-Chip Devices , Phospholipids/chemistry , Polymers/chemistry , Surface Properties
8.
Sleep Med ; 80: 204-209, 2021 04.
Article in English | MEDLINE | ID: mdl-33607555

ABSTRACT

OBJECTIVE: This study sought to validate the Japanese version of the Sleep Hygiene Practices Scale (SHPS-J). PATIENTS/METHODS: A cross-sectional questionnaire-based study was conducted via the internet. In total, 854 participants (435 men, 419 women; mean age, 42.91 ± 11.54 years) were asked to complete all scales, and 283 of them were asked to complete the same scales two weeks later. The survey consisted of the SHPS-J, the Japanese version of the Insomnia Severity Index (ISI-J), and the Japanese version of the Pittsburgh Sleep Quality Index (PSQI-J). The SHPS-J was developed according to the International Society for Pharmacoeconomics and Outcomes Research Task Force for Translation and Cultural Adaption. For the analysis, participants were divided into three groups: insomnia syndrome, insomnia symptoms, and good sleep groups. RESULTS: The SHPS-J had good test-retest reliability (ICC: 0.55-0.76) and adequate internal consistency (α = 0.54-0.74), except with regard to eating/drinking behaviors. The factorial validity of the four-factor structure was confirmed through a confirmatory factor analysis; however, one item related to eating/drinking behaviors had no significant factor loading. The construct validity was confirmed through a correlation analysis between each domain of the SHPS-J and ISI-J (r = 0.19-0.60, p < 0.01). The results of clinical validation confirmed that all domains of the SHPS-J were significantly higher for individuals with insomnia than for good sleepers. CONCLUSIONS: This study confirmed both the reliability and validity of the SHPS-J.


Subject(s)
Sleep Hygiene , Sleep Initiation and Maintenance Disorders , Adult , Cross-Sectional Studies , Female , Humans , Japan , Male , Middle Aged , Psychometrics , Reproducibility of Results , Sleep , Sleep Initiation and Maintenance Disorders/diagnosis , Surveys and Questionnaires
9.
Curr Microbiol ; 78(2): 789-795, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33389060

ABSTRACT

A novel bacterial strain, designated MHJ-10JT, was isolated from a soil sample obtained from a grassland in Inner Mongolia, China. MHJ-10JT strain could grow at 4-37 °C (optimum: 30 °C) and pH 4-9 (optimum: pH 6), as well as in the presence of 0-6% NaCl (optimum: 1%). Cells of strain MHJ-10JT are Gram-negative, rod-shaped, and motile. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain MHJ-10JT was most closely related to Pseudomonas lutea OK2T (98.5% 16S rRNA gene sequence similarity). The values of the average nucleotide identities (ANI) and digital DNA-DNA hybridization (dDDH) between strain MHJ-10JT and its related species were all below 80.5% and 24.4%, respectively, which are significantly lower than the thresholds of 95% for ANI and 70% for DDH for species delineation. The genomic G + C content of the MHJ-10JT strain is 64.8 mol%. Based on the phenotypic, genotypic, chemotaxonomic, and phylogenetic analyses, strain MHJ-10JT can be assigned to the genus Pseudomonas. In this study, we propose that strain MHJ-10JT be classified as a novel species belonging to the genus Pseudomonas with the species name Pseudomonas pratensis sp. nov. The type strain of the proposed novel species is MHJ-10JT (= KCTC 82206T = CGMCC 17322T).


Subject(s)
Soil Microbiology , Soil , Bacterial Typing Techniques , China , DNA, Bacterial/genetics , Fatty Acids/analysis , Grassland , Phospholipids/analysis , Phylogeny , Pseudomonas/genetics , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
10.
Microbes Environ ; 35(1)2020.
Article in English | MEDLINE | ID: mdl-31932539

ABSTRACT

Diverse members of Bradyrhizobium diazoefficiens, B. japonicum, and B. ottawaense were isolated from the roots of field-grown sorghum plants in Fukushima, and classified into "Rhizobia" with nodulated soybeans, "Free-living diazotrophs", and "Non-diazotrophs" by nitrogen fixation and nodulation assays. Genome analyses revealed that B. ottawaense members possessed genes for N2O reduction, but lacked those for the Type VI secretion system (T6SS). T6SS is a new bacterial weapon against microbial competitors. Since T6SS-possessing B. diazoefficiens and B. japonicum have mainly been isolated from soybean nodules in Japan, T6SS-lacking B. ottawaense members may be a cryptic lineage of soybean bradyrhizobia in Japan.


Subject(s)
Biodiversity , Bradyrhizobium/genetics , Oxidoreductases/genetics , Sorghum/microbiology , Type VI Secretion Systems/deficiency , Bradyrhizobium/classification , Bradyrhizobium/isolation & purification , Genetic Variation , Nitrogen Fixation/genetics , Phylogeny , Plant Root Nodulation/genetics , Plant Roots/microbiology , Rhizobium/classification , Rhizobium/genetics , Rhizobium/isolation & purification , Type VI Secretion Systems/genetics
11.
Knee Surg Sports Traumatol Arthrosc ; 28(6): 1750-1756, 2020 Jun.
Article in English | MEDLINE | ID: mdl-31250057

ABSTRACT

PURPOSE: The purpose of the present study was to use arthroscopy to evaluate the effect of distal tuberosity osteotomy (DTO) in open-wedge high tibial osteotomy (OW-HTO) on patellofemoral (PF) cartilage degradation. METHODS: Between 2012 and 2017, 46 knees underwent DTO in OW-HTO, and 65 knees underwent conventional OW-HTO (cOW-HTO). To assess changes in patellar height, the Blackburne-Peel (BP) ratio and the Caton-Deschamps (CD) index were measured. Arthroscopic evaluation on the PF joint was performed at the initial osteotomy and at the second-look procedure 1 year later. Statistical analyses were performed to compare difference between the DTO and the cOW-HTO group. RESULTS: In the cOW-HTO group, the mean BP ratio and CD index decreased significantly from 0.81 and 0.89 preoperatively, respectively, to 0.69 and 0.76 postoperatively, respectively (p < 0.001). In contrast, the DTO group maintained a consistent patellar height; the mean BP ratio and CD index were 0.77 and 0.83 preoperatively, respectively, and 0.73 and 0.80 postoperatively, respectively. Upon arthroscopic evaluation, 39 of 46 patients (84.8%) in the DTO group showed no progression of PF cartilage degradation at the second look; indeed, five of 46 patients (10.9%) even demonstrated improvement. In contrast, 21 of 65 patients (32.3%) in the cOW-HTO group exhibited increased PF cartilage degradation. There was a significant difference in progression of PF cartilage degradation between DTO and cOW-HTO (p < 0.001). CONCLUSION: DTO in OW-HTO maintained the preoperative patellar height, which could help prevent progression of cartilage degeneration in the PF joint after surgery. In respect of the biplanar osteotomy direction in OW-HTO, the DTO, rather than cOWHTO, is the preferred technique for the treatment of varus knee osteoarthritis to avoid progression of PF cartilage degradation. LEVEL OF EVIDENCE: III.


Subject(s)
Arthroscopy/methods , Cartilage Diseases/surgery , Cartilage/physiopathology , Cartilage/surgery , Osteoarthritis, Knee/surgery , Osteotomy/methods , Aged , Disease Progression , Female , Humans , Knee Joint/surgery , Male , Middle Aged , Patella/surgery , Patellofemoral Joint/surgery , Postoperative Period , Preoperative Period , Second-Look Surgery , Tibia/surgery
12.
Microbes Environ ; 34(4): 446-450, 2019 Dec 27.
Article in English | MEDLINE | ID: mdl-31413227

ABSTRACT

Plant-associated bacteria are critical for plant growth and health. However, the effects of plant growth stages on the bacterial community remain unclear. Analyses of the microbiome associated with field-grown soybean revealed a marked shift in the bacterial community during the growth stages. The relative abundance of Methylorubrum in the leaf and stem increased from 0.2% to more than 45%, but decreased to approximately 15%, with a peak at the flowering stage at which nitrogen metabolism changed in the soybean plant. These results suggest the significance of a time-series analysis for understanding the relationship between the microbial community and host plant physiology.


Subject(s)
Glycine max/growth & development , Glycine max/microbiology , Methylobacteriaceae/growth & development , Microbiota , Bacteria/classification , Bacteria/genetics , Bacteria/growth & development , DNA, Bacterial/genetics , Methylobacteriaceae/classification , Methylobacteriaceae/genetics , Nitrogen/metabolism , Plant Leaves/growth & development , Plant Leaves/microbiology , RNA, Ribosomal, 16S/genetics , Glycine max/metabolism
13.
Tohoku J Exp Med ; 248(3): 209-216, 2019 07.
Article in English | MEDLINE | ID: mdl-31366819

ABSTRACT

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by systemic joint inflammation and may manifest as interstitial pneumonia (IP). Methotrexate (MTX) is one of the main therapeutic drugs used for RA, but MTX could cause severe side effects, including Pneumocystis jirovecii pneumonia (PCP) and IP. Owing to similar symptoms, it is sometimes difficult to discriminate MTX therapy-associated PCP (MTX-PCP) and MTX therapy-associated IP (MTX-IP). Soluble interleukin-2 receptor (sIL-2R) is considered a marker of T-cell activation, and serum sIL-2R levels are elevated in RA and PCP. This led us to hypothesize that serum sIL-2R is a potential biomarker for discriminating MTX-PCP and MTX-IP. Accordingly, we carried out a retrospective analysis of 20 MITX-PCP cases, 30 MTX-IP cases, and as controls, 16 patients with RA-associated IP (RA-IP) and 13 patients with PCP without MTX treatment (PCP group). C-reactive protein and alveolar-arterial oxygen differences were higher in the MTX-PCP group than those in the RA-IP and MTX-IP groups. Importantly, serum levels of sIL-2R in MTX-PCP were significantly higher than those in other three groups. Based on the receiver operating characteristic curve, the cut-off level of sIL-2R resulting in the highest diagnostic accuracy for MTX-PCP was 1,311.5 U/mL, discriminating between MTX-PCP and other groups with 91.7% sensitivity and 78.6% specificity. Thus, patients with MTX-PCP show a higher degree of systemic inflammation, severe hypoxemia, and increased sIL-2R levels compared with those in MTX-IP cases. In conclusion, serum sIL-2R could be a biomarker for PCP diagnosis among patients with RA under MTX therapy.


Subject(s)
Arthritis, Rheumatoid/complications , Arthritis, Rheumatoid/drug therapy , Methotrexate/therapeutic use , Pneumocystis carinii/physiology , Pneumonia/blood , Pneumonia/complications , Receptors, Interleukin-2/blood , Aged , Arthritis, Rheumatoid/diagnostic imaging , Biomarkers/blood , Female , Humans , Male , Middle Aged , Pneumonia/diagnostic imaging , Pneumonia/microbiology , ROC Curve , Solubility , Tomography, X-Ray Computed
14.
Inflamm Intest Dis ; 4(1): 35-40, 2019 May.
Article in English | MEDLINE | ID: mdl-31172011

ABSTRACT

A 40-year-old man with refractory ulcerative colitis (UC) was treated with tumor necrosis factor α inhibitor (anti-TNFα), infliximab. One month later, the chest computed tomography and laboratory test showed noninfectious interstitial lung disease (ILD) and elevation of serum Krebs von den Lungen-6 (KL-6). Fortunately, ILD disappeared after the discontinuation with anti-TNFα. Two and a half years after his first UC treatment, he was treated again with another anti-TNFα, adalimumab, for relapse and he had a second ILD. This course suggested anti-TNFα induced ILD. The characteristics of anti-TNFα-induced ILD in inflammatory bowel disease (IBD) are not well understood. We summarized and investigated the characteristics of such patients based on a literature review including 15 cases. It suggested that anti-TNFα-induced ILD in IBD might be rare and tends to have a better outcome compared with ILD in rheumatoid arthritis.

15.
Front Microbiol ; 10: 407, 2019.
Article in English | MEDLINE | ID: mdl-30915047

ABSTRACT

Sorghum (Sorghum bicolor) is cultivated worldwide for food, bioethanol, and fodder production. Although nitrogen fixation in sorghum has been studied since the 1970s, N2-fixing bacteria have not been widely examined in field-grown sorghum plants because the identification of functional diazotrophs depends on the culture method used. The aim of this study was to identify functional N2-fixing bacteria associated with field-grown sorghum by using "omics" approaches. Four lines of sorghum (KM1, KM2, KM4, and KM5) were grown in a field in Fukushima, Japan. The nitrogen-fixing activities of the roots, leaves, and stems were evaluated by acetylene reduction and 15N2-feeding assays. The highest nitrogen-fixing activities were detected in the roots of lines KM1 and KM2 at the late growth stage. Bacterial cells extracted from KM1 and KM2 roots were analyzed by metagenome, proteome, and isolation approaches and their DNA was isolated and sequenced. Nitrogenase structural gene sequences in the metagenome sequences were retrieved using two nitrogenase databases. Most sequences were assigned to nifHDK of Bradyrhizobium species, including non-nodulating Bradyrhizobium sp. S23321 and photosynthetic B. oligotrophicum S58T. Amplicon sequence and metagenome analysis revealed a relatively higher abundance (2.9-3.6%) of Bradyrhizobium in the roots. Proteome analysis indicated that three NifHDK proteins of Bradyrhizobium species were consistently detected across sample replicates. By using oligotrophic media, we purified eight bradyrhizobial isolates. Among them, two bradyrhizobial isolates possessed 16S rRNA and nif genes similar to those in S23321 and S58T which were predicted as functional diazotrophs by omics approaches. Both free-living cells of the isolates expressed N2-fixing activity in a semi-solid medium according to an acetylene reduction assay. These results suggest that major functional N2-fixing bacteria in sorghum roots are unique bradyrhizobia that resemble photosynthetic B. oligotrophicum S58T and non-nodulating Bradyrhizobium sp. S23321. Based on our findings, we discuss the N2-fixing activity level of sorghum plants, phylogenetic and genomic comparison with diazotrophic bacteria in other crops, and Bradyrhizobium diversity in N2 fixation and nodulation.

16.
J Infect Chemother ; 25(6): 480-484, 2019 Jun.
Article in English | MEDLINE | ID: mdl-30824300

ABSTRACT

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging tick-borne disease caused by a novel bunyavirus. The mechanism underlying disease progression remains unknown, and effective treatment strategy for SFTS is yet to be completely established, making its increasing incidence and subsequent mortality a great concern. Here, we present the autopsy case of a patient with rapidly progressed, fatal SFTS infection. Her viral titer and serum cytokines levels were measured daily and compared with the values of a survivor of the infection. Our findings elucidate the clinical features and pathophysiology of SFTS.


Subject(s)
Bunyaviridae Infections/diagnosis , Cytokines/blood , Phlebovirus/isolation & purification , Tick-Borne Diseases/diagnosis , Viral Load , Aged , Bunyaviridae Infections/blood , Bunyaviridae Infections/immunology , Cadaver , Cytokines/immunology , Fatal Outcome , Female , Humans , Phlebovirus/immunology , Prognosis , Tick-Borne Diseases/blood , Tick-Borne Diseases/immunology
17.
ACS Nano ; 13(3): 2795-2803, 2019 Mar 26.
Article in English | MEDLINE | ID: mdl-30626184

ABSTRACT

The large-area fabrication of aligned nanopatterns with sub-5 nm feature size is crucial for developing nanodevices. Highly ordered nanostructures fabricated through molecular self-assembly exhibit substantial potential for sub-5 nm patterning techniques. Previously, we had reported the fabrication of silica nanogrooves with sub-5 nm periodicity on a Si substrate by using the outermost surfaces of cylindrical surfactant micelles as a template. However, uniaxial alignment of nanogrooves on the entire substrate surface has not yet been achieved. In this study, uniaxially aligned silica nanogrooves were prepared on the entire surface of a Si substrate (2 cm × 2 cm) by utilizing a poly(dimethylsiloxane) (PDMS) stamp with a striped pattern. The PDMS stamp was placed on the surface of a surfactant thin film precoated on the substrate, although the stamp was not in direct contact with the substrate as in the case of the soft nanoimprint technique. The substrate was then exposed to water vapor, during which cylindrical micelles were aligned in the direction of the guide. Subsequently, by exposing the substrate to an NH3-water vapor mixture, the outermost surfaces of the aligned micelles facing the substrate were replicated with soluble silicate species. The formation of uniaxially aligned nanogrooves on the entire surface of the centimeter-scale substrate was verified by scanning electron microscopy observations and grazing-incidence small-angle X-ray scattering analysis. Thus, herein we provide a simple large-area fabrication method for uniaxially aligned nanopatterns with ultrafine pitch.

18.
Ther Deliv ; 9(10): 711-729, 2018 10.
Article in English | MEDLINE | ID: mdl-30277135

ABSTRACT

AIM: Tacrolimus (TAC) is an important drug for inflammatory diseases. However, TAC has several limitations, such as variable trough concentrations among individuals and a high medication frequency. In this study, we created NK61060, a novel micellar TAC formulation, to circumvent these disadvantages. MATERIALS & METHODS: Immunosuppressive activity of NK61060 was determined in the collagen-induced arthritis rat model, mannan-induced arthritis mouse model and dextran sodium sulfate-induced colitis mouse model. The pharmacokinetics and toxicology of NK61060 were evaluated in those models. RESULTS: In arthritis and colitis models, NK61060 exhibited superior immunosuppressive activity compared with that of TAC. Pharmacokinetic and toxicological analyses indicated that NK61060 had a wider safety margin and could be administered at a reduced medication frequency. CONCLUSION: NK61060 mitigates the trough concentration variability and the medication frequency and it may be a safer and more effective option for use in clinical settings. Further studies are needed to determine its clinical usefulness.


Subject(s)
Arthritis, Experimental/drug therapy , Arthritis, Rheumatoid/drug therapy , Colitis, Ulcerative/drug therapy , Drug Carriers/chemistry , Immunosuppressive Agents/administration & dosage , Tacrolimus/administration & dosage , Animals , Area Under Curve , Arthritis, Experimental/immunology , Arthritis, Rheumatoid/immunology , Colitis, Ulcerative/chemically induced , Colitis, Ulcerative/immunology , Collagen/immunology , Dextran Sulfate/toxicity , Drug Administration Schedule , Drug Evaluation, Preclinical , Female , Humans , Mannans/immunology , Mice , Mice, Inbred ICR , Micelles , Polyethylene Glycols/chemistry , Rats , Rats, Sprague-Dawley
19.
Langmuir ; 34(4): 1733-1741, 2018 01 30.
Article in English | MEDLINE | ID: mdl-29272588

ABSTRACT

The flexible control of nanopatterns by a bottom-up process at the nanometer scale is essential for nanofabrication with a finer pitch. We have previously reported that for the fabrication of linear nanopatterns with sub-5 nm periodicity on Si substrates the outermost surfaces of assembled micelles facing the substrates can be replicated with soluble silicate species generated from the Si substrates under basic conditions. In this study, concentrically arranged nanogrooves with a sub-5 nm periodicity were prepared on Si substrates by replicating the outermost surfaces of bent micelles guided by silica particles. The Si substrates, where silica particles and surfactants films were deposited, were exposed to an NH3-water vapor mixture. During the vapor treatment, cylindrical micelles became arranged in concentric patterns centered on the silica particles, and their outermost surfaces facing the substrates were replicated by soluble silicate species on the Si substrates. The thinness of the surfactant film on the substrate is crucial for the formation of concentric silica nanogrooves because the out-of-plane orientations of the micelles are suppressed at the interface. Surprisingly, the domains of the concentric silica nanogrooves spread to much larger areas than the maximum cross-sectional areas of the particles, and the size of the domains increased linearly with the radii of the particles. The extension of concentric nanogrooves is discussed on the basis of the orientational elastic energies of the micelles around one silica particle. This study of the formation of bent nanogrooves guided by the outlines of readily deposited nanoscale objects provides a new nanostructure-guiding process.

20.
Microbes Environ ; 32(4): 390-393, 2017 Dec 27.
Article in English | MEDLINE | ID: mdl-29109334

ABSTRACT

Archaeal communities in mineral soils were compared between a boreal forest in Finland and cold-temperate forest in Japan using 16S rRNA gene-targeted high-throughput sequencing. In boreal soils, Thaumarchaeota Group 1.1c archaea predominated and Thaumarchaeota Group 1.1a-associated and Group 1.1b archaea were also detected. In temperate soils, Thaumarchaeota Group 1.1a-associated and Group 1.1b archaea were dominant members at the subsurface, whereas their dominancy was replaced by Thermoplasmata archaea at the subsoil. An analysis of the ammonia monooxygenase subunit A gene of Archaea also indicated the distribution of Thaumarchaeota Group 1.1a-associated and Group 1.1b archaea in these soils.


Subject(s)
Euryarchaeota , Microbiota/genetics , Soil Microbiology , Euryarchaeota/classification , Euryarchaeota/genetics , Euryarchaeota/isolation & purification , Finland , High-Throughput Nucleotide Sequencing , Japan , Oxidoreductases/genetics , RNA, Ribosomal, 16S/genetics , Soil/chemistry , Taiga
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