Publications by authors named "Shiori Yamamoto"

31 Publications

Bacterial Distribution and Community Structure in Beef Cattle Liver and Bile at Slaughter.

J Food Prot 2021 Nov 24. Epub 2021 Nov 24.

National Institute of Health Sciences 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki, JAPAN Kanagawa 210-9501 +81442706563.

In this study, the distribution of hygienic indicator bacteria in cattle livers and bile was examined at slaughterhouses. First, 127 cattle livers with gallbladders were carefully eviscerated from the carcasses at 10 slaughterhouses. Microbiological examination showed that 9 bile (7.1%) and 19 liver parenchyma (15.0%) samples were positive for the family Enterobacteriaceae (EB) with means ± SD of 3.68 ± 4.63 log CFU/mL and 1.59 ± 2.47 log CFU/g, respectively; thus, bacterial contamination was apparent even at the postevisceration stage. Subsequently, 70 cattle livers were obtained at the postprocessing/storage stage from 7 of the ten slaughterhouses; microbiological analysis revealed greater means of EB in the liver parenchyma (means ± SD of 3.00 ± 3.89 log CFU/g, P =0.011) than those at postevisceration stage, suggesting that bacterial dissemination and/or replication occurred in the liver parenchyma during processing and storage. According to 16S rRNA ion semiconductor sequencing analysis of representative samples from 12 cattle, Proteobacteria , Firmicutes , and Actinobacteria were dominant in both the parenchyma and bile, in which EB/ Escherichia coli were predominate among EB-rich livers. These results suggest that bile plays a role as a vehicle for bacterial transmission to the liver parenchyma. This is the first study to demonstrate bacterial distribution and community structure in the liver and biliary microecosystem of cattle at slaughter. Our data provide possible implication of EB testing in bile to screen cattle livers contaminated with high levels of fecal indicator bacteria.
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http://dx.doi.org/10.4315/JFP-21-288DOI Listing
November 2021

Zfhx4 regulates endochondral ossification as the transcriptional platform of Osterix in mice.

Commun Biol 2021 11 3;4(1):1258. Epub 2021 Nov 3.

Department of Molecular and Cellular Biochemistry, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka, 565-0871, Japan.

Endochondral ossification is regulated by transcription factors that include SRY-box transcription factor 9, runt-related protein 2 (Runx2), and Osterix. However, the sequential and harmonious regulation of the multiple steps of endochondral ossification is unclear. This study identified zinc finger homeodomain 4 (Zfhx4) as a crucial transcriptional partner of Osterix. We found that Zfhx4 was highly expressed in cartilage and that Zfhx4 deficient mice had reduced expression of matrix metallopeptidase 13 and inhibited calcification of cartilage matrices. These phenotypes were very similar to impaired chondrogenesis in Osterix deficient mice. Coimmunoprecipitation and immunofluorescence indicated a physical interaction between Zfhx4 and Osterix. Notably, Zfhx4 and Osterix double mutant mice showed more severe phenotype than Zfhx4 deficient mice. Additionally, Zfhx4 interacted with Runx2 that functions upstream of Osterix. Our findings suggest that Zfhx4 coordinates the transcriptional network of Osterix and, consequently, endochondral ossification.
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http://dx.doi.org/10.1038/s42003-021-02793-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8566502PMC
November 2021

Long-Term Grow-Out Affects Colonization Fitness in Coincidence With Altered Microbiota and Lipid Composition in the Cecum of Laying Hens.

Front Vet Sci 2021 18;8:675570. Epub 2021 Jun 18.

Department of Animal Hygiene, Tokyo University of Agriculture, Atsugi City, Japan.

is one of the leading causes of gastrointestinal illness worldwide and is mainly transmitted from chicken through the food chain. Previous studies have provided increasing evidence that this pathogen can colonize and replicate in broiler chicken during its breeding; however, its temporal kinetics in laying hen are poorly understood. Considering the possible interaction between and gut microbiota, the current study was conducted to address the temporal dynamics of in the cecum of laying hen over 40 weeks, with possible alteration of the gut microbiota and fatty acid (FA) components. Following oral infection with 81-176, inocula were stably recovered from ceca for up to 8 weeks post-infection (.). From 16 weeks ., most birds became negative for and remained negative up to 40 weeks . 16S rRNA gene sequencing analyses revealed that most of the altered relative rRNA gene abundances occurred in the order , in which increased relative rRNA gene abundances were observed at >16 weeks . in the families , and . Lipidome analyses revealed increased levels of sterols associated with bile acid metabolisms in the cecum at 16 and/or 24 weeks . compared with those detected at 8 weeks ., suggesting that altered microbiota and bile acid metabolism might underlie the decreased colonization fitness of in the gut of laying hens.
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http://dx.doi.org/10.3389/fvets.2021.675570DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8249580PMC
June 2021

Untargeted Phylogenetic Group III of Multi-drug-Resistant Bacillus cereus Isolated Using Fraser Medium from Retail Chickens in Ho Chi Minh City.

Curr Microbiol 2021 Aug 26;78(8):3115-3123. Epub 2021 Jun 26.

Faculty of Contemporary Human Life Science, Tezukayama University, Nara, Japan.

The prevalence of food-borne bacteria in developing countries is less well understood than in developed countries. The ISO11290-1 isolation method is commonly used to study Listeria contamination in chicken; however, all isolates are identified as untargeted Bacillus cereus. This study aimed to determine the classification, antibiotic susceptibility, and virulence genes of B. cereus isolated from retail chickens in Vietnam. Bacterial isolation using the ISO11290-1 method yielded 12 strains of B. cereus from seven out of 60 chickens. For determining bacterial diversity, panC and multilocus sequence typing (MLST) analyses were performed. PanC analysis showed that all seven strains belong to the phylogenetic group III, to which the highest risk of foodborne illnesses was associated. MLST analysis showed that most strains contained a ST205 complex; further, all strains were found to be resistant to ampicillin, ciprofloxacin, and tetracycline. Virulence genes were also investigated. ces, a cereulide-related gene, was detected in 50% of the isolated strains, followed by cytK, nheA, and hblA enterotoxins in 41.7%, 16.7%, and 25% of the strains, respectively. In conclusion, B. cereus may be erroneously detected when attempting to detect Listeria in food using the ISO11290-1 method. Further study of the prevalence of B. cereus in Vietnamese food is needed to improve food safety.
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http://dx.doi.org/10.1007/s00284-021-02562-1DOI Listing
August 2021

Effects of a surface prereacted glass-ionomer filler coating material on biofilm formation and inhibition of dentin demineralization.

Clin Oral Investig 2021 Feb 23;25(2):683-690. Epub 2020 Sep 23.

Department of Cariology and Operative Dentistry, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo, 113-8549, Japan.

Objectives: This study investigated the ability of a surface prereacted glass-ionomer (S-PRG) coating material to inhibit the biofilm formation and demineralization of dentin.

Methods And Materials: Dentin specimens were randomly divided into three groups: (1) no coating (control), (2) S-PRG filler-containing coat, and (3) a nonS-PRG filler-containing coat. Streptococcus mutans biofilms were grown on the dentin surfaces in a microcosm for 20 h. Then, the quantity of bacteria and water-insoluble glucan in the retained biofilm on the dentin surface were measured. Regarding demineralization inhibition test, specimens were demineralized for 5 days then sectioned into halves and observed under confocal laser scanning microscope (CLSM). One-way ANOVA and Tukey's HSD were used for statistical analysis.

Results: The estimated mean surface roughness for specimens in the S-PRG group was statistically significantly higher than the estimates for both the nonS-PRG and the control group specimens. The quantity of bacteria and water-insoluble glucan/mm revealed that the S-PRG group prevented biofilm formation and bacterial adhesion to the dentin surface compared with the control and nonS-PRG groups. The S-PRG group recorded the highest acid-resistance ability with no surface loss.

Conclusion: Application of S-PRG barrier coat on dentin surfaces can inhibit biofilm formation as well as protecting the dentin surface against demineralization.

Clinical Significance: Coating material containing S-PRG fillers might be used for caries prevention, through inhibiting biofilm formation, enhancing mineralization, and reducing acidic attack by cariogenic bacteria.
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http://dx.doi.org/10.1007/s00784-020-03577-xDOI Listing
February 2021

Draft Genome Sequences of Non-HS-Producing Strains of Salmonella enterica Serovars Infantis, Enteritidis, Berta, and Kiambu in Japan.

Microbiol Resour Announc 2020 Jul 23;9(30). Epub 2020 Jul 23.

Department of Microbiology, Tokyo University of Agriculture, Tokyo, Japan.

We report the draft genome sequences of six strains of serovars Berta, Enteritidis, Infantis, and Kiambu, isolated from humans or chicken meats in Osaka, Japan, that were negative for hydrogen sulfide production. Their genome sizes ranged from 4,460,389 to 4,933,483 bp, with 3 to 9 rRNAs and 64 to 73 tRNAs and with coverages of 95× to 159×.
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http://dx.doi.org/10.1128/MRA.00335-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7378023PMC
July 2020

Plasmid Sequences of Four Large Plasmids Carrying Antimicrobial Resistance Genes in Escherichia coli Strains Isolated from Beef Cattle in Japan.

Microbiol Resour Announc 2020 May 14;9(20). Epub 2020 May 14.

Applied Microbiology, Graduate School of Agriculture, Hokkaido University, Sapporo, Hokkaido, Japan.

is a common reservoir for antimicrobial resistance genes that can be easily transformed to possess multidrug resistance through plasmid transfer. To understand multidrug resistance plasmids, we report the plasmid sequences of four large plasmids carrying a number of genes related to antimicrobial resistance that were found in strains isolated from beef cattle.
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http://dx.doi.org/10.1128/MRA.00219-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7225533PMC
May 2020

Draft Genome Sequence of Stenotrophomonas maltophilia CRB139-1, Isolated from Poultry Meat in Japan.

Microbiol Resour Announc 2020 Mar 19;9(12). Epub 2020 Mar 19.

Division of Biomedical Food Research, National Institute of Health Sciences, Kawasaki-ku, Kawasaki, Kanagawa, Japan.

is a nosocomial pathogen that primarily causes respiratory infection in humans. This pathogen is widely distributed in the environment, including in foods. Here, we report the draft genome sequence of strain CRB139-1, isolated from poultry meat in Japan. The genome size was 4,619,918 bp at 90× coverage.
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http://dx.doi.org/10.1128/MRA.00075-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7082454PMC
March 2020

Phylogenetic Diversity and Antimicrobial Resistance of Campylobacter coli from Humans and Animals in Japan.

Microbes Environ 2019 Jun 21;34(2):146-154. Epub 2019 Mar 21.

Department of Animal Hygiene, Tokyo University of Agriculture.

The phylogenetic diversity and antimicrobial resistance (AMR) of Campylobacter coli from humans and animals in Japan between 2008 and 2014 were investigated. A total of 338 foodborne campylobacterioses were reported in Osaka, and C. coli was isolated from 38 cases (11.2%). In the present study, 119 C. coli strains (42 from humans, 25 each from poultry, cattle, and swine, and 2 from wild mallard) were examined by multilocus sequence typing (MLST). MLST assigned 36 sequence types (STs), including 14 novel STs; all human strains and 91% of animal strains (70/77) were assigned to the ST-828 clonal complex. The predominant human ST was ST-860 (18/42, 43%), followed by ST-1068 (8/42, 19%); these STs were also predominant in poultry (ST-860, 9/25, 36%) and cattle (ST-1068, 18/25, 72%). ST-1562 was only predominant in swine (11/25, 44.0%). Swine strains showed the greatest resistance to erythromycin (EM; 92.0%), while EM resistance was only found in 2 out of the 42 human strains examined (4.8%). All EM-resistant swine strains (n=15) exhibited a common point mutation in the 23S rRNA sequence (A2085G), and the tetO gene was detected in 22 out of the 23 TET-resistant swine strains. A whole genome sequencing analysis of four representative swine ST-1562 strains revealed abundant AMR-associated gene clusters in their genomes, suggesting horizontal gene transfer events during host adaptation. This is the first study to demonstrate the phylogenetic diversity and AMR profiles of C. coli in Japan. The present results suggest that poultry and cattle are major reservoirs, improving our knowledge on the epidemiological and ecological traits of this pathogen.
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http://dx.doi.org/10.1264/jsme2.ME18115DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6594732PMC
June 2019

Hoxa13 regulates expression of common Hox target genes involved in cartilage development to coordinate the expansion of the autopodal anlage.

Dev Growth Differ 2019 Apr 20;61(3):228-251. Epub 2019 Mar 20.

Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya-shi, Aichi-ken, Japan.

To elucidate the role of Hox genes in limb cartilage development, we identified the target genes of HOXA11 and HOXA13 by ChIP-Seq. The ChIP DNA fragment contained evolutionarily conserved sequences and multiple highly conserved HOX binding sites. A substantial portion of the HOXA11 ChIP fragment overlapped with the HOXA13 ChIP fragment indicating that both factors share common targets. Deletion of the target regions neighboring Bmp2 or Tshz2 reduced their expression in the autopod suggesting that they function as the limb bud-specific enhancers. We identified the Hox downstream genes as exhibiting expression changes in the Hoxa13 knock out (KO) and Hoxd11-13 deletion double mutant (Hox13 dKO) autopod by Genechip analysis. The Hox downstream genes neighboring the ChIP fragment were defined as the direct targets of Hox. We analyzed the spatial expression pattern of the Hox target genes that encode two different categories of transcription factors during autopod development and Hox13dKO limb bud. (a) Bcl11a, encoding a repressor of cartilage differentiation, was expressed in the E11.5 autopod and was substantially reduced in the Hox13dKO. (b) The transcription factors Aff3, Bnc2, Nfib and Runx1t1 were expressed in the zeugopodal cartilage but not in the autopod due to the repressive or relatively weak transcriptional activity of Hox13 at E11.5. Interestingly, the expression of these genes was later observed in the autopodal cartilage at E12.5. These results indicate that Hox13 transiently suspends the cartilage differentiation in the autopodal anlage via multiple pathways until establishing the paddle-shaped structure required to generate five digits.
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http://dx.doi.org/10.1111/dgd.12601DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6850407PMC
April 2019

Investigation of morphological changes for the discrimination of nucleated red blood cells and other leukocytes in Sysmex XN hematology analyzer scattergrams using transmission electron microscopy.

Pract Lab Med 2017 Aug 10;8:70-76. Epub 2017 May 10.

Scientific Research, Scientific Affairs, Sysmex Corporation, 1-3-2 Murotani, Nishi-ku, Kobe 651-2241, Japan.

Background: The WNR channel of the XN-Series automated hematology analyzer (Sysmex) counts white blood cells (WBCs) and simultaneously performs a differential counting of basophils and nucleated red blood cells (NRBCs). The detection process involves exposing the cells to WNR-specific reagents containing an acidic detergent and a fluorescent dye and measuring the intensity of the forward scattered light (FSC) and side fluorescence light (SFL).

Method: We treated isolated peripheral WBCs and NRBCs with specific reagents and assessed the morphological changes in NRBCs and each leukocyte type using transmission electron microscopy (TEM).

Results: The results from a flow cytometer (FCM) showed that, after exposure to the reagents, basophils appeared on the highest FSC and SFL areas compared to other leukocytes on the WNR scattergram. Owing to the hemolysis of reticulocytes and erythrocytes, NRBCs that survived the reagent treatment could be distinguished by their lower intensity than those of the other leukocytes on the WNR scattergram. We investigated the significance of the relationship between the TEM and FCM results after the reagent treatment.

Conclusion: We confirmed that the WNR channel differentiates the blood cells on the WNR scattergram based on differences in the amount of residual cytoplasm and nucleic acids.
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http://dx.doi.org/10.1016/j.plabm.2017.05.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5575374PMC
August 2017

Genome Sequence of Strain Adk2012 Associated with a Foodborne Botulinum Case in Tottori, Japan, in 2012.

Genome Announc 2017 Aug 24;5(34). Epub 2017 Aug 24.

Department of Bacteriology II, National Institute of Infectious Diseases, Tokyo, Japan.

We report here a draft genome sequence of Adk2012 responsible for a foodborne botulism case that occurred in Tottori, Japan, in 2012. Its genome size was 2,904,173 bp, with 46 rRNAs and 54 tRNAs, at a coverage of 14.5×.
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http://dx.doi.org/10.1128/genomeA.00872-17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5571422PMC
August 2017

Draft Genome Sequence of CAM970 and CAM962, Associated with a Large Outbreak of Foodborne Illness in Fukuoka, Japan, in 2016.

Genome Announc 2017 Jun 15;5(24). Epub 2017 Jun 15.

Health Science Section, Fukuoka City Institute of Health and Environment, Fukuoka, Japan.

Here, we report the draft genome sequences of CAM970 and CAM962, which were associated with a large outbreak of foodborne illness originating from undercooked chicken sushi in Fukuoka, Japan, in May 2016. Their genome sizes were 1,690,901 and 1,704,736 bp, with 22 and 23 rRNAs, 9 and 9 tRNAs, and 411× and 419× coverage for CAM970 and CAM962, respectively.
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http://dx.doi.org/10.1128/genomeA.00508-17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5473265PMC
June 2017

Draft Genome Sequence of Five Shiga Toxin-Producing Strains Isolated from Wild Deer in Japan.

Genome Announc 2017 Mar 2;5(9). Epub 2017 Mar 2.

Laboratory of Animal Hygiene, Kitasato University, Aomori, Japan.

Shiga toxin-producing (STEC) is one of the major foodborne pathogens. Having observed the wide distribution of this pathogen in wild deer, we report here the draft genome sequence of five STEC strains isolated from wild deer () in Hokkaido, Japan.
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http://dx.doi.org/10.1128/genomeA.01455-16DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5334574PMC
March 2017

Iron-chelating agent, deferasirox, inhibits neutrophil activation and extracellular trap formation.

Clin Exp Pharmacol Physiol 2016 10;43(10):915-20

Kobe University Hospital, Kobe, Japan.

Iron-chelating agents, which are frequently prescribed to transfusion-dependent patients, have various useful biological effects in addition to chelation. Reactive oxygen species (ROS) produced by neutrophils can cause pulmonary endothelial cell damage, which can lead to acute lung injury (ALI). We previously reported that deferasirox (DFS), an iron-chelating agent, inhibits phorbol myristate acetate (PMA) or formyl-methionyl-leucyl-phenylalanine (fMLP)-induced ROS production in neutrophils, in vitro. Here, we investigate whether DFS inhibits vacuolization in neutrophils and neutrophil extracellular trap (NET) formation. Human neutrophils were incubated with DFS and stimulated with PMA or fMLP. Human neutrophils were separated from heparinized peripheral blood using density gradient centrifugation, and subsequently incubated with DFS. After 10 minutes, neutrophils were stimulated by PMA or fMLP. Vacuole formation was observed by electron microscopy. For observing NET formations using microscopes, immunohistological analyses using citrullinated histone H3 and myeloperoxidase antibodies, and SYTOX Green (an impermeable DNA detection dye) staining, were conducted. NET formation was measured as the quantity of double-stranded DNA (dsDNA), using the AccuBlue Broad Range dsDNA Quantitation Kit. DFS (50 μmol/L) inhibited vacuole formation in the cytoplasm and NET formation. Additionally, 5-100 μmol/L concentration of DFS inhibited the release of dsDNA in a dose-independent manner. We demonstrate that DFS inhibits not only ROS production but also vacuolization and NET formation in neutrophils. These results suggest the possibility of protective effects of DFS against NET-related adverse effects, including ALI and thrombosis.
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http://dx.doi.org/10.1111/1440-1681.12612DOI Listing
October 2016

A role for HOX13 proteins in the regulatory switch between TADs at the HoxD locus.

Genes Dev 2016 05 19;30(10):1172-86. Epub 2016 May 19.

Department of Genetics and Evolution, University of Geneva, 1211 Geneva 4, Switzerland; School of Life Sciences, Federal Institute of Technology, Lausanne, 1015 Lausanne, Switzerland;

During vertebrate limb development, Hoxd genes are regulated following a bimodal strategy involving two topologically associating domains (TADs) located on either side of the gene cluster. These regulatory landscapes alternatively control different subsets of Hoxd targets, first into the arm and subsequently into the digits. We studied the transition between these two global regulations, a switch that correlates with the positioning of the wrist, which articulates these two main limb segments. We show that the HOX13 proteins themselves help switch off the telomeric TAD, likely through a global repressive mechanism. At the same time, they directly interact with distal enhancers to sustain the activity of the centromeric TAD, thus explaining both the sequential and exclusive operating processes of these two regulatory domains. We propose a model in which the activation of Hox13 gene expression in distal limb cells both interrupts the proximal Hox gene regulation and re-enforces the distal regulation. In the absence of HOX13 proteins, a proximal limb structure grows without any sign of wrist articulation, likely related to an ancestral fish-like condition.
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http://dx.doi.org/10.1101/gad.281055.116DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4888838PMC
May 2016

Eosinophil extracellular trap cell death-derived DNA traps: Their presence in secretions and functional attributes.

J Allergy Clin Immunol 2016 Jan 9;137(1):258-267. Epub 2015 Jun 9.

Divisions of Allergy and Inflammation and Infectious Diseases, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Mass.

Background: Activated human eosinophils, as well as neutrophils, can release extracellular chromatin to form DNA traps through cytolytic extracellular trap cell death (ETosis). Although formations of neutrophil DNA traps are recognized in patients with various inflammatory conditions, neither the presence of ETosis-derived eosinophil DNA traps in human allergic diseases nor the characteristics of these DNA traps have been studied.

Objective: We investigated the presence of ETosis-derived DNA traps in eosinophil-rich sinus and ear secretions and the functional attributes of ETosis DNA traps.

Methods: Eosinophil-rich secretions obtained from patients with eosinophilic chronic rhinosinusitis and eosinophilic otitis media were studied microscopically. In vitro studies of ETosis and DNA trap formation used blood-derived eosinophils and neutrophils, and studies of the binding capacities of DNA traps used labeled bacteria and fluorescent microbeads. Stabilities of DNA traps were evaluated by using fluorescence microscopy.

Results: Abundant nuclear histone H1-bearing DNA traps formed in vivo in the eosinophilic secretions and contributed to their increased viscosity. In vitro, after brief shear flow, eosinophil ETosis-elicited DNA traps assembled to form stable aggregates. Eosinophil DNA traps entrapped bacteria and fungi and, through hydrophobic interactions, microbeads. In comparison with neutrophil-derived DNA traps, eosinophil DNA traps ultrastructurally exhibited thicker fibers with globular structures and were less susceptible to leukocyte-derived proteolytic degradation, likely because of the lesser protease activities of eosinophils.

Conclusions: In human allergic diseases local cytolysis of eosinophils not only releases free eosinophil granules but also generates nuclear-derived DNA traps that are major extracellular structural components within eosinophil-rich secretions.
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http://dx.doi.org/10.1016/j.jaci.2015.04.041DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4674385PMC
January 2016

Comparison of optical data from flow cytometry and microscopy of leukocytes after exposure to specific reagents.

Microscopy (Oxf) 2015 Oct 25;64(5):305-10. Epub 2015 May 25.

Scientific Research, Scientific Affairs, Sysmex Corporation, 1-3-2 Murotani, Nishi-ku, Kobe 651-2241, Japan.

Flow cytometry and microscopy are equally important in cell analysis. However, few reports have compared the optical data (cell size, internal complexity and fluorescent signal) from flow cytometry and microscopy. In this study, we compared the scattergram from XN-series, a flow cytometry based hematology analyzer with microscopic images of similarly treated leukocytes, and investigated the correlation between the appearance in the scattergram and cell size, internal complexity and fluorescence intensity. Healthy human peripheral blood was analyzed using the XN analyzer. For microscopic comparison, five types of leukocytes (monocytes, lymphocytes, basophils, neutrophils and eosinophils) were isolated from the peripheral blood by centrifugation and magnetic cell sorting, treated with a specific reagent and analyzed using electron microscopy, laser microscopy and confocal laser microscopy. Cell size, residual internal structures and fluorescence intensity correlated with intensity of forward-scattering, side scattering and fluorescent light. In this study, optical data from a clinically used hematology analyzer was clarified using microscopic images.
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http://dx.doi.org/10.1093/jmicro/dfv023DOI Listing
October 2015

Exploratory study of pre-surgical medications with dienogest or leuprorelin in laparoscopic cystectomy of endometrial cysts.

J Obstet Gynaecol Res 2015 Aug 1;41(8):1234-9. Epub 2015 Apr 1.

Department of Obstetrics and Gynecology, Gifu University School of Medicine.

Aim: The aim of this study was to compare the effects of pre-surgical medication with dienogest or leuprorelin on post-surgical ovarian function.

Material And Methods: We conducted an exploratory study in two centers in Japan that comprised 30 patients with ovarian endometrial cysts for whom surgical excision was planned. Patients were enrolled and divided into pre-surgical medication groups with dienogest or leuprorelin for 12 weeks. Thereafter, patients were treated by laparoscopic cystectomy. The primary outcome was ovarian function post-surgery, as assessed by serum anti-Müllerian hormone (AMH) level, antral follicle count (AFC) and resumption of menses. Secondary outcome was the effect of pre-surgical medication, as assessed by the size of endometrial cysts and visual analog scale (VAS) score. Serum AMH, AFC, size of endometrial cysts, and VAS scores were measured at baseline (before medication), after medication (1 day before surgery), and at 4 and 12 weeks post-surgery.

Results: Serum AMH levels did not change after pre-surgical medication with either dienogest or leuprorelin. Although AMH decreased after surgery, it recovered by 12 weeks post-surgery in both groups with no statistically significant difference. Mean AFC did not change after surgery in either group. Menses returned by 12 weeks post-surgery in all patients except for those who were pregnant. The rate of reduction of endometrial cyst volume did not differ between the groups. Both dienogest and leuprorelin were associated with substantial reductions in VAS scores.

Conclusion: There were no statistically significant differences between pre-surgical medication with dienogest and leuprorelin in post-surgical ovarian function. Both medications were effective in reducing endometrial cyst volume and VAS score.
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http://dx.doi.org/10.1111/jog.12701DOI Listing
August 2015

Leucophores are similar to xanthophores in their specification and differentiation processes in medaka.

Proc Natl Acad Sci U S A 2014 May 6;111(20):7343-8. Epub 2014 May 6.

Interuniversity Bio-Backup Project Center, National Institute for Basic Biology, Okazaki 444-8787, Aichi, Japan;Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies (SOKENDAI), Okazaki, Aichi 444-8787, Japan;Laboratory of Bioresources, National Institute for Basic Biology, Okazaki 444-8585, Aichi, Japan.

Animal body color is generated primarily by neural crest-derived pigment cells in the skin. Mammals and birds have only melanocytes on the surface of their bodies; however, fish have a variety of pigment cell types or chromatophores, including melanophores, xanthophores, and iridophores. The medaka has a unique chromatophore type called the leucophore. The genetic basis of chromatophore diversity remains poorly understood. Here, we report that three loci in medaka, namely, leucophore free (lf), lf-2, and white leucophore (wl), which affect leucophore and xanthophore differentiation, encode solute carrier family 2, member 15b (slc2a15b), paired box gene 7a (pax7a), and solute carrier family 2 facilitated glucose transporter, member 11b (slc2a11b), respectively. Because lf-2, a loss-of-function mutant for pax7a, causes defects in the formation of xanthophore and leucophore precursor cells, pax7a is critical for the development of the chromatophores. This genetic evidence implies that leucophores are similar to xanthophores, although it was previously thought that leucophores were related to iridophores, as these chromatophores have purine-dependent light reflection. Our identification of slc2a15b and slc2a11b as genes critical for the differentiation of leucophores and xanthophores in medaka led to a further finding that the existence of these two genes in the genome coincides with the presence of xanthophores in nonmammalian vertebrates: birds have yellow-pigmented irises with xanthophore-like intracellular organelles. Our findings provide clues for revealing diverse evolutionary mechanisms of pigment cell formation in animals.
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http://dx.doi.org/10.1073/pnas.1311254111DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4034200PMC
May 2014

Characterization of multi-antibiotic-resistant Escherichia coli Isolated from beef cattle in Japan.

Microbes Environ 2014 30;29(2):136-44. Epub 2014 Apr 30.

Applied Microbiology, Graduate School of Agriculture, Hokkaido University.

The emergence of multiple-antibiotic-resistance bacteria is increasing, which is a particular concern on livestock farms. We previously isolated 1,347 antimicrobial-resistant (AMR) Escherichia coli strains from the feces of beef cattle on 14 Japanese farms. In the present study, the genetic backgrounds and phylogenetic relationships of 45 AMR isolates were characterized by the chromosome phylotype, AMR phenotype, AMR genotype, and plasmid type. These isolates were classified into five chromosome phylotypes, which were closely linked to the farms from which they were isolated, suggesting that each farm had its own E. coli phylotype. AMR phenotype and plasmid type analyses yielded 8 and 14 types, all of which were associated with the chromosomal phylotype and, thus, to the original farms. AMR genotype analysis revealed more variety, with 16 types, indicating both inter- and intra-farm diversity. Different phylotype isolates from the same farm shared highly similar plasmid types, which indicated that plasmids with AMR genes could be transferred between phylotypes, thereby generating multi-antibiotic-resistant microorganisms. This ecological study demonstrated that the chromosome phylotype was strongly correlated with the farm from which they were isolated, while the AMR phenotype, genotype, and plasmid type were generally correlated with the chromosome phylotype and farm source.
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http://dx.doi.org/10.1264/jsme2.me13173DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4103519PMC
February 2017

Prevalence and characteristics of Listeria monocytogenes in feces of black beef cattle reared in three geographically distant areas in Japan.

Foodborne Pathog Dis 2014 Feb 1;11(2):96-103. Epub 2013 Nov 1.

1 Department of Nutrition, School of Nursing and Nutrition, Tenshi College , Sapporo, Hokkaido, Japan .

This study was conducted to determine the prevalence and characteristics of Listeria monocytogenes in the feces of black beef cattle reared in geographically distant areas in Japan. We surveyed 130 farms in the following three areas: northern (Hokkaido prefecture), central (Gifu and Mie prefectures), and southern (Oita, Miyazaki, and Kagoshima prefectures) areas and collected 1738 fecal samples. Our data showed the following isolation rate for each area: northern, 11.4% of 651; central, 2.8% of 572; and southern, 2.9% of 515, indicating that the isolation rate in the northern area was significantly higher than that in the central or southern areas (p<0.01). Moreover, serotyping of 996 isolates identified 1/2b as the most prevalent serotype (40.5%), followed by 1/2a (36.9%), 4b (21.6%), and 4ab (1.0%). In the northern area, multiple serotypes were isolated from 60% of L. monocytogenes-positive farms. In addition, multiple serotypes were isolated from individual fecal samples from 18 cattle. Pulsed-field gel electrophoresis (PFGE) characterization of 239 isolates detected 48 different PFGE types. We found that isolates from northern farms were genetically diverse compared to those from central and southern farms. Five isolates from human clinical cases and three isolates from animal clinical cases were identical to isolates from black beef cattle. Furthermore, the isolates from northern and central farms were characterized to possess epidemic clone II or III markers. We next showed that the isolates were susceptible to penicillin, ampicillin, amoxicillin, gentamicin, kanamycin, streptomycin, erythromycin, vancomycin, tetracycline, chloramphenicol, ciprofloxacin, and trimethoprim/sulfamethoxazole. Taken together, our survey provides crucial data regarding the prevalence and characteristics of L. monocytogenes in black beef cattle farms throughout Japan.
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http://dx.doi.org/10.1089/fpd.2013.1616DOI Listing
February 2014

Prevalence and molecular epidemiological characterization of antimicrobial-resistant Escherichia coli isolates from Japanese black beef cattle.

J Food Prot 2013 Mar;76(3):394-404

Department of Nutrition, School of Nursing and Nutrition, Tenshi College, Higashi-ku, Sapporo, Hokkaido 065-0013, Japan.

We investigated the prevalence of antimicrobial-resistant Escherichia coli in Japanese black beef cattle from the three major production regions of Japan. We collected and examined 291 fecal samples from Japanese black beef cattle in Hokkaido, Chubu, and Kyushu. Of the 3,147 E. coli isolates, 1,397 (44.4%) were resistant to one or more antibiotics; these included 553 (39.8%) of 1,388 isolates from Hokkaido, 352 (54.4%) of 647 isolates from Chubu, and 492 (44.2%) of 1,112 isolates from Kyushu. The difference in resistance rates between the three regions was significant. The antibiotics with the highest rates of resistance were oxytetracycline and dihydrostreptomycin (35.8% each), followed by ampicillin (21.4%). Further, E. coli isolates from calves had higher resistance rates than those from growing cattle and mature cattle, and the calf isolates showed high rates of resistance to gentamicin (20.2%), enrofloxacin (9.4%), and ceftiofur (4.2%). In addition, the high degrees of similarity in the genotypes of the isolates and in the resistance patterns on each farm suggest that resistance bacteria and resistance genes were horizontally transferred. Most isolates, in each of the three regions, harbored resistance genes such as blaTEM, strA, strB, aphA1, aphAI-IAB, and catI. In contrast to the isolates from Kyushu, most of which harbored aacC2, tetB, and dfrA12, the isolates from Hokkaido and Chubu harbored a variety of resistance genes. Furthermore, the prevalence of genes for resistance to dihydrostreptomycin, gentamicin, chloramphenicol, and trimethoprim differed significantly between the regions. This is the first large-scale study describing and comparing antimicrobial-resistant bacteria from different regions in Japan. The results will contribute to improving food safety and promoting careful usage of antimicrobial agents.
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http://dx.doi.org/10.4315/0362-028X.JFP-12-273DOI Listing
March 2013

Prevalence and characteristics of Listeria monocytogenes in bovine colostrum in Japan.

J Food Prot 2013 Feb;76(2):248-55

Department of Nutrition, School of Nursing and Nutrition, Tenshi College, Higashi-ku, Sapporo, Hokkaido 065-0013, Japan.

This study was conducted to determine the prevalence and characteristics of Listeria monocytogenes in bovine colostrum in Japan. We collected bovine colostrum samples from 210 dams from 21 dairy farms in Hokkaido prefecture (Japan) between March and June 2009. L. monocytogenes was detected in samples from 6 (28.6%) of the 21 farms. Of the 210 samples, 16 (7.6%) were positive for L. monocytogenes. We recovered 80 L. monocytogenes isolates; 44 (55%) isolates were classified as serotype 1/2b and 36 (45%) were classified as serotype 4b. The isolates were susceptible to penicillin, ampicillin, amoxicillin, gentamicin, kanamycin, streptomycin, erythromycin, vancomycin, tetracycline, chloramphenicol, ciprofloxacin, and trimethoprim-sulfamethoxazole. Pulsed-field gel electrophoresis (PFGE) characterization of the 80 isolates revealed six PFGE types. Two PFGE types corresponded to human listeriosis cases. Most L. monocytogenes isolates possessed virulence-associated genes (actA, hly, iap, inlA, inlC, mpl, plcA, plcB, opuCA, prfA, and clpC). One PFGE type isolate possessed an epidemic clone II marker. Our findings suggest that isolates from bovine colostrum have the potential to cause human and animal listeriosis. This is the first study on the prevalence and characteristics of L. monocytogenes isolated from bovine colostrum obtained from dairy farms. Our results have important implications for improving public health and elucidating the epidemiology of L. monocytogenes in bovine colostrum.
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http://dx.doi.org/10.4315/0362-028X.JFP-12-278DOI Listing
February 2013

Prevalence of Salmonella isolates and antimicrobial resistance patterns in chicken meat throughout Japan.

J Food Prot 2011 Feb;74(2):270-3

Department of Nutrition, School of Nursing and Nutrition, Tenshi College, Sapporo, Hokkaido 065-0013, Japan.

We investigated the prevalence of Salmonella in chicken meat from northern, central, and southern Japan. Between 2006 and 2008, 821 samples from these three regions were collected and examined. Salmonella isolates were detected in 164 (20.0%) of these samples, with 15 (10.0%) of 150, 113 (27.5%) of 411, and 36 (13.8%) of 260 recovered from the northern, central, and southern regions, respectively. We recovered 452 Salmonella isolates. From the isolates, 27 serovars were identified; the predominant serovars isolated were Salmonella Infantis (n=81), Salmonella Kalamu (n=56), and Salmonella Schwarzengrund (n=43). Of the 452 isolates, 443 (98.0%) were resistant to one or more antibiotics, and 221 (48.9%) showed multiple-antibiotic resistance, thereby implying that multiple-antibiotic resistant Salmonella organisms are widespread in chicken meat in Japan. Resistance to oxytetracycline was most common (72.6%), followed by dihydrostreptomycin (69.2%) and bicozamycin (49.1%). This study, the first to report Salmonella prevalence in chicken meat throughout Japan, could provide valuable data for monitoring and controlling Salmonella infection in the future.
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http://dx.doi.org/10.4315/0362-028X.JFP-10-215DOI Listing
February 2011

Mice deficient for glucagon gene-derived peptides display normoglycemia and hyperplasia of islet {alpha}-cells but not of intestinal L-cells.

Mol Endocrinol 2009 Dec 9;23(12):1990-9. Epub 2009 Oct 9.

Department of Genetics, Research Institute of Environmental Medicine, Nagoya University, Japan.

Multiple bioactive peptides, including glucagon, glucagon-like peptide-1 (GLP-1), and GLP-2, are derived from the glucagon gene (Gcg). In the present study, we disrupted Gcg by introduction of GFP cDNA and established a knock-in mouse line. Gcg(gfp/gfp) mice that lack most, if not all, of Gcg-derived peptides were born in an expected Mendelian ratio without gross abnormalities. Gcg(gfp/gfp) mice showed lower blood glucose levels at 2 wk of age, but those in adult Gcg(gfp/gfp) mice were not significantly different from those in Gcg(+/+) and Gcg(gfp/+) mice, even after starvation for 16 h. Serum insulin levels in Gcg(gfp/gfp) mice were lower than in Gcg(+/+) and Gcg(gfp/+) on ad libitum feeding, but no significant differences were observed on starvation. Islet alpha-cells and intestinal L-cells were readily visualized in Gcg(gfp/gfp) and Gcg(gfp/+) mice under fluorescence. The Gcg(gfp/gfp) postnatally developed hyperplasia of islet alpha-cells, whereas the population of intestinal L-cells was not increased. In the Gcg(gfp/gfp), expression of Aristaless-related homeobox (Arx) was markedly increased in pancreas but not in intestine and suggested involvement of Arx in differential regulation of proliferation of Gcg-expressing cells. These results illustrated that Gcg-derived peptides are dispensable for survival and maintaining normoglycemia in adult mice and that Gcg-derived peptides differentially regulate proliferation/differentiation of alpha-cells and L-cells. The present model is useful for analyzing glucose/energy metabolism in the absence of Gcg-derived peptides. It is useful also for analysis of the development, differentiation, and function of Gcg-expressing cells, because such cells are readily visualized by fluorescence in this model.
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http://dx.doi.org/10.1210/me.2009-0296DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5419124PMC
December 2009

Risk factors for major limb amputations in diabetic foot gangrene patients.

Diabetes Res Clin Pract 2006 Mar 31;71(3):272-9. Epub 2005 Aug 31.

Department of Dermatology, Osaka Koseinenkin Hospital, 4-2-78, Fukushima-ku, Fukushima, Osaka 553-0003, Japan.

We analyzed the clinical picture of diabetic foot lesion patients to investigate the risk factors for major limb amputations. The subjects were 210 diabetic foot lesion patients treated at our department over the past 9 years. The mean follow-up period was 604.5 [standard deviation (S.D.) 451.2] days with a median value of 492 days. There were 113 men and 97 women. By the final follow-up day, 18 underwent bypass surgeries (9%) and 13 skin grafts (6%), while 110 patients (52%) finally required limb amputation. The breakdown was 45 major amputations above or below the knee and 65 minor amputations of the toes or metatarsals. The outcomes of the major amputations were retrospectively analyzed by group. The blood glucose control was poor in all 45 major amputees, and their mean HbA1c (8.80%) was higher than that in the minor or non-amputation group (7.79%, P = 0.035). In the major amputation group, two patients had loss of vision due to retinopathy, and 30 patients received long-term hemodialysis due to nephropathy. The rate of arteriosclerosis obliterans (ASO) was higher in the major amputation group than in the minor or non-amputation group, and arteriography showed significantly high rates of multiple stenosis. Multivariate analysis of these results by the proportional hazard model showed that ASO with multiple stenosis (hazard ratio 3.23, 95% CI: 1.12-5.10), hemodialysis (2.14, 95% CI: 1.17-3.44), and HbA1c (1.20, 95% CI: 1.03-1.41) were independent risk factors for major amputation. The 3-year survival rate was 24.1% in the major amputation group and 93.0% in the minor or non-amputation group, and the life expectancy was significantly lower for the major amputees than the minor or non-amputees (P<0.0001). Together with early detection and treatment of foot lesions, good blood glucose control and early management of systemic complications such as nephropathy and arteriosclerosis are considered important to avoid major amputations.
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http://dx.doi.org/10.1016/j.diabres.2005.07.005DOI Listing
March 2006

Cyclic tetramers composed of rhodium(III), iridium(III), or ruthenium(II) half-sandwich and 6-purinethiones.

Inorg Chem 2002 Dec;41(25):6824-30

Department of Chemistry, Graduate School of Science, Osaka University, 1-16 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan.

Six new cyclic tetranuclear complexes [[M(Cp*)(L)](4)](4+) and [[Ru(II)(L)(cymene)](4)](4+) [Cp* = eta(5)-C(5)Me(5), cymene = eta(6)-p-MeC(6)H(4)Pr(i); M = Rh(III) and Ir(III); HL = 6-purinethione (H(2)put) and 2-amino-6-purinethione (H(2)aput)] were prepared in a self-assembly manner and characterized by NMR spectroscopy, electrospray ionization mass spectrometry, and X-ray crystal structure analysis. The two crystal structures of [[Rh(Cp*)(H(0.5)put)](4)](CF(3)SO(3))(2) and [[Ir(Cp*)(Haput)](4)](CF(3)SO(3))(4) revealed that they have similar S(4) structures with an alternate chirality array of CACA, and all ligands adopt a mu-1kappaN(9):2kappa(2)S(6),N(7) coordination mode. The orientations of the four bridging ligands are alternately up and down, and they form a central square cavity. Interestingly, the cationic tetramers of the former are stacked up along the c axis, resulting in an infinite channel-like cavity. The driving force of this stacking is due to intermolecular double hydrogen bonds [N(1)-H...N(21) = 2.752(4) A] at both sides of the cavity. In the two Rh(III)- and Ru(II)-H(2)aput systems, it turned out that the dimeric species are dominantly formed in the reaction solutions but finally convert into the tetrameric species.
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http://dx.doi.org/10.1021/ic020449oDOI Listing
December 2002

Cyclic Hexamer with a Cubic Cavity: Crystal Structure of

Angew Chem Int Ed Engl 2001 Jun;40(12):2268-2271

Department of Chemistry, Graduate School of Science Osaka University 1-16 Machikaneyama-cho, Toyonaka, Osaka 560-0043 (Japan).

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June 2001
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