Publications by authors named "Yuya Nagasawa"

20 Publications

  • Page 1 of 1

Intramammary infection caused by Staphylococcus aureus increases IgA antibodies to iron-regulated surface determinant-A, -B, and -H in bovine milk.

Vet Immunol Immunopathol 2021 May 31;235:110235. Epub 2021 Mar 31.

Dairy Hygiene Research Division, National Institute of Animal Health, National Agriculture and Food Research Organization, 4 Hitsujigaoka, Toyohira, Sapporo, Hokkaido, 062-0045, Japan. Electronic address:

The aim of this study was to identify virulence factors that have high immunogenicity. An in vivo-expressed Staphylococcus aureus antigen was identified by probing bacteriophage expression libraries of S. aureus with antibodies in bovine mastitis milk. Eighteen clones were isolated, and their proteins were identified as 5 characterised proteins (IsdA, Protein A, IsdB, autolysin, and imidazole glycerol phosphate dehydratase) and 13 hypothetical proteins. We focused on IsdA, IsdB, and IsdH as virulence factors that have a high immunogenicity and are capable of inducing a specific humoral immune response in S. aureus-infected quarters. The optical density (OD) values of IsdA and IsdB IgA and IgG antibodies in milk affected by naturally occurring mastitis caused by S. aureus increased significantly compared to those in healthy milk. In the experimental infection study, the OD values of IsdA- and B-specific IgA and IgG antibodies were significantly increased from 2 to 4 weeks after S. aureus infection compared to day 0 (P < 0.05). On the other hand, we demonstrated that milk from natural and experimental intramammary infections caused by S. aureus are associated with significantly higher IgA levels against IsdH (P < 0.05), but no significant change in IgG levels. Our findings facilitated our understanding of the pathogenicity of S. aureus in bovine mastitis, as well as the mechanisms by which specific humoral immune responses to S. aureus infection are induced. In addition, the results obtained could provide insight into how bovine mastitis can be controlled, for example, through vaccination.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.vetimm.2021.110235DOI Listing
May 2021

Rapid Detection From Clinical Mastitis Milk by Colloidal Gold Nanoparticle-Based Immunochromatographic Strips.

Front Vet Sci 2019 22;6:504. Epub 2020 Jan 22.

Dairy Hygiene Unit, Division of Pathology and Pathophysiology, Hokkaido Research Station, National Institute of Animal Health, National Agriculture and Food Research Organization, Sapporo, Japan.

Rapid diagnostic technologies for bovine mastitis caused by () are urgently needed. In the current study, we generated an anti-ribosomal protein-L7/L12 antibody to detect and an anti-ribosomal protein-L7/L12 antibody-coated immune-chromatographic strip (ICS) test. Moreover, we determined the ability of the ICS test to detect from milk samples collected from cows with clinical mastitis. The developed ICS reacted to in a bacteria load-dependent manner with a detection limit of ~10 CFU/mL. In the evaluation of possible cross-reactivity of the ICS test, six strains of coagulase-negative Staphylococci showed slightly positive reactions, although at a lower level; however, other bacteria were completely negative. Next, we investigated the sensitivity and specificity of the ICS test compared with the bacteriological culture method using milk samples from clinical bovine mastitis. The results of the experiments demonstrated that the ICS test had high sensitivity [100%, 95% confidence interval (CI): 91.3-100%] and specificity (91.9%, CI: 90.5-91.9%) compared with culture tests. In addition, the kappa statistic demonstrated that ICS tests showed substantial agreement (k = 0.77, CI: 0.66-0.87) with culture tests. Positive correlations were observed for the statistical analysis between ( gene) copy numbers and ICS test scores in mastitic milk infected by . Therefore, we assume that this new detection method using ICS may be useful as a highly sensitive -screening method for the diagnosis of bovine mastitis. Our findings support the ongoing effort to develop an ICS method for bovine -induced mastitis, which can contribute to the rapid diagnosis of this disease.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fvets.2019.00504DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6988510PMC
January 2020

An Incremental Sit-to-Stand Exercise for Evaluating Physical Capacity in Older Patients with Type 2 Diabetes.

Tohoku J Exp Med 2019 12;249(4):241-248

Department of Internal Medicine, Matsumoto City Hospital.

Exercise is recommended for older patients with type 2 diabetes mellitus (T2DM), and increased physical activity contributes to better management of their condition. The conventional exercise test with treadmill or cycle ergometer (CE) for assessing physical capacity, such as peak oxygen uptake (VO) and anaerobic threshold (AT), is not always usable for older patients with T2DM. The incremental sit-to-stand (ISTS) exercise is an incremental exercise test using external signals to control the sit-to-stand rate in a given time frame and can be performed in a small space using only a chair. This study aimed to examine the validity of the physical capacity assessment during the ISTS exercise, based on the relationships between the ISTS performance, peak VO, AT on ISTS exercise and CE test, in older patients with T2DM. Twenty-two patients with T2DM (10 men, 12 women; mean age, 68.0 years; range, 61-77 years) performed ISTS exercise (according to an existing protocol) and CE test in a randomized manner. Peak VO, AT, and completion time were determined for the ISTS exercise and CE test. Peak VO during ISTS exercise was significantly associated with that during the CE test (r = 0.89, p < 0.01). The completion time on the ISTS exercise was significantly associated with peak VO (r = 0.80, p < 0.01) and AT on the ISTS exercise (r = 0.78, p < 0.01). The ISTS exercise is a useful tool to determine the physical capacity and estimate peak VO and AT in older people with T2DM.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1620/tjem.249.241DOI Listing
December 2019

Staphylococcus aureus-specific IgA antibody in milk suppresses the multiplication of S. aureus in infected bovine udder.

BMC Vet Res 2019 Aug 9;15(1):286. Epub 2019 Aug 9.

Dairy Hygiene Unit, Division of Pathology and Pathophysiology, Hokkaido Research Station, National Institute of Animal Health, National Agriculture and Food Research Organization, 4 Hitsujigaoka, Toyohira, Sapporo, Hokkaido, 062-0045, Japan.

Background: Bovine mastitis caused by Staphylococcus aureus (S. aureus) is extremely difficult to control and new methods for its prevention and management are required. Nasal vaccines may prevent initial bovine mastitis infection caused by S. aureus. However, limited information is available regarding induction of mucosal immune response through nasal immunization with antigen and its suppression of S. aureus multiplication during bovine mastitis. This study sought to investigate whether induction of immunoglobulin A (IgA) in milk by nasal immunization could suppress multiplication of S. aureus in the bovine udder.

Results: Nasal immunization with formalin-killed S. aureus conjugated with a cationic cholesteryl-group-bearing pullulan-nanogel was performed. Anti-S. aureus-specific IgA antibodies were significantly more abundant in the milk of immunized cows than in non-immunized animals (P < 0.05). S. aureus counts in the quarter were negative in both non-immunized and nasal-immunized cows 1 week after mock infusion. In S. aureus-infused quarters, S. aureus multiplication was significantly suppressed in immunized compared with non-immunized cows (P < 0.05). Furthermore, a significant negative correlation was found between S. aureus-specific IgA antibodies and S. aureus counts in infused quarters of both non-immunized and nasal-immunized cows (r = - 0.811, P < 0.01).

Conclusion: In conclusion, the present study demonstrates that S. aureus-specific IgA antibodies in milk successfully suppressed the multiplication of S. aureus in infected bovine udders. Although the exact mechanism explaining such suppressive effect remains to be elucidated, nasal vaccines that can induce humoral immunity may help prevent initial infection with S. aureus and the onset of bovine mastitis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s12917-019-2025-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6688226PMC
August 2019

Factors associated with functional rehabilitation outcomes of non-operative treatment for hip fractures: a retrospective study.

J Phys Ther Sci 2019 May 10;31(5):453-456. Epub 2019 May 10.

Department of Orthopedic Surgery, Matsumoto City Hospital, Japan.

[Purpose] Limited data are available regarding the outcomes of non-operative treatment for hip fractures. We investigated the factors associated with functional rehabilitation outcomes in patients undergoing non-operative treatment for hip fractures. [Participants and Methods] We investigated 57 patients with hip fractures who underwent non-operative treatment. We retrospectively analyzed medical or rehabilitation outcomes and functional outcomes (assessed using the Functional Independence Measure tool). We examined the association between functional outcomes and other factors and compared the medical and rehabilitation outcomes between mobile and immobile patients at the time of discharge. [Results] Of the 57 patients investigated, 15 (26.3%) were mobile at discharge. We observed a significant association between the Functional Independence Measure subscores (Motor and Cognitive) and serum albumin levels. Serum albumin levels and the Functional Independence Measure subscores (Motor and Cognitive) were significantly higher in mobile than in immobile patients. [Conclusion] We observed that functional outcomes at discharge in patients undergoing non-operative treatment for hip fractures were associated with serum albumin ratios and the Functional Independence Measure-Cognitive score.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1589/jpts.31.453DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6511504PMC
May 2019

Validity and reproducibility of an incremental sit-to-stand exercise test in healthy middle-aged individuals.

J Phys Ther Sci 2019 May 10;31(5):414-417. Epub 2019 May 10.

Physical Therapy, School of Health Sciences, Shinshu University, Japan.

[Purpose] We aimed to evaluate the validity and reproducibility of the incremental sit-to-stand exercise test for aerobic fitness evaluation in healthy middle-aged individuals. [Participants and Methods] Thirteen healthy middle-aged individuals randomly underwent the incremental sit-to-stand exercise and cycle ergometer tests, and the peak oxygen uptake was measured during both tests. Pearson's correlation coefficients were used to assess the strength of the association between the peak oxygen uptake measured during the aforementioned tests. Intraclass correlation coefficients with 95% confidence intervals for peak oxygen uptake obtained during the first, second, and third incremental sit-to-stand exercise tests were used to determine the reproducibility of this test. [Results] The peak oxygen uptake measured during the incremental sit-to-stand exercise test was strongly associated with that measured during the cycle ergometer test (r=0.86). The intraclass correlation coefficients (95% confidence intervals) used to verify the association of the peak oxygen uptake between the first and the second incremental sit-to-stand exercise tests and between the second and third incremental sit-to-stand exercise tests were 0.92 (0.66-0.99) and 0.96 (0.82-0.99), respectively. [Conclusion] The incremental sit-to-stand exercise test is a valid and reproducible tool to evaluate aerobic fitness in healthy middle-aged individuals.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1589/jpts.31.414DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6511505PMC
May 2019

The bacterial load in milk is associated with clinical severity in cases of bovine coliform mastitis.

J Vet Med Sci 2019 Jan 26;81(1):107-112. Epub 2018 Nov 26.

Dairy Hygiene Unit, Division of Pathology and Pathophysiology, Hokkaido Research Station, National Institute of Animal Health, National Agriculture and Food Research Organization, 4 Hitsujigaoka, Toyohira, Sapporo, Hokkaido 062-0045, Japan.

We evaluated the relationship between the severity of coliform mastitis and bacterial load in 106 quarter milk samples. We found no significant relationship between somatic cell count and coliform bacterial load in milk in bovine clinical coliform mastitis. Results of the Cochran-Armitage test for trend in milk bacterial load proportions indicated a significant decreasing low group (P<0.001), increasing medium group (P<0.002) and increasing high group (P<0.02) with increasing clinical grade. The present study indicates that the coliform bacterial load in milk is significantly associated with clinical severity states in cases of bovine coliform mastitis, and can be a useful indicator for optimal management of this disease.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1292/jvms.18-0581DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6361639PMC
January 2019

Energetics and electronic structures of perylene confined in carbon nanotubes.

R Soc Open Sci 2018 Jun 27;5(6):180359. Epub 2018 Jun 27.

Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8571, Japan.

The energetics and geometries of perylene encapsulated in carbon nanotubes (CNTs) have been investigated employing density functional theory using the generalized gradient approximation combined with the van der Waals correction. Our calculations show that the encapsulated perylene molecules possess two metastable molecular conformations with respect to the CNT wall, which are almost degenerate with each other. A standing conformation, with respect to the CNT wall, is the ground state conformation for a semiconducting (19,0)CNT, while a lying conformation is the ground state for a metallic (11,11)CNT. Cooperation and competition between perylene-perylene and perylene-CNT interactions cause these possible perylene conformations inside CNTs. However, the electronic structure of the CNT encapsulating the perylene molecules is found to be insensitive to the molecular conformation.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1098/rsos.180359DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6030348PMC
June 2018

Phenotypic and functional analysis of bovine peripheral blood dendritic cells before parturition by a novel purification method.

Anim Sci J 2018 Jul 30;89(7):1011-1019. Epub 2018 Apr 30.

Cellular Biology Laboratory, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.

Dendritic cells (DCs) are specialized antigen presenting cells specializing in antigen uptake and processing, and play an important role in the innate and adaptive immune response. A subset of bovine peripheral blood DCs was identified as CD172a /CD11c /MHC (major histocompatibility complex) class II cells. Although DCs are identified at 0.1%-0.7% of peripheral blood mononuclear cells (PBMC), the phenotype and function of DCs remain poorly understood with regard to maintaining tolerance during the pregnancy. All cattle used in this study were 1 month before parturition. We have established a novel method for the purification of DCs from PBMC using magnetic-activated cell sorting, and purified the CD172a /CD11c DCs, with high expression of MHC class II and CD40, at 84.8% purity. There were individual differences in the expressions of CD205 and co-stimulatory molecules CD80 and CD86 on DCs. There were positive correlations between expression of cytokine and co-stimulatory molecules in DCs, and the DCs maintained their immune tolerance, evidenced by their low expressions of the co-stimulatory molecules and cytokine production. These results suggest that before parturition a half of DCs may be immature and tend to maintain tolerance based on the low cytokine production, and the other DCs with high co-stimulatory molecules may already have the ability of modulating the T-cell linage.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/asj.13014DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6055732PMC
July 2018

Identification of a novel mechanism of action of bovine IgG antibodies specific for Staphylococcus aureus.

Vet Res 2018 02 26;49(1):22. Epub 2018 Feb 26.

International Education and Research Center for Food and Agricultural Immunology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, 980-0845, Japan.

Staphylococcus aureus is a major pathogen that causes subclinical mastitis associated with huge economic losses to the dairy industry. A few vaccines for bovine mastitis are available, and they are expected to induce the production of S. aureus-specific antibodies that prevent bacterial adherence to host cells or promote opsonization by phagocytes. However, the efficacy of such vaccines are still under debate; therefore, further research focusing on improving the current vaccines by seeking additional mechanisms of action is required to reduce economic losses due to mastitis in the dairy industry. Here, we generated S. aureus-specific bovine IgG antibodies (anti-S. aureus) that directly inhibited bacterial growth in vitro. Inhibition depended on specificity for anti-S. aureus, not the interaction between Protein A and the fragment crystallizable region of the IgG antibodies or bacterial agglutination. An in vitro culture study using S. aureus strain JE2 and its deletion mutant JE2ΔSrtA, which lacks the gene encoding sortase A, revealed that the effect of anti-S. aureus was sortase-A-independent. Sortase A is involved in the synthesis of cell-wall-associated proteins. Thus, other surface molecules, such as membrane proteins, cell surface polysaccharides, or both, may trigger the inhibition of bacterial growth by anti-S. aureus. Together, our findings contribute insights into developing new strategies to further improve the available mastitis vaccine by designing a novel antigen on the surface of S. aureus to induce inhibitory signals that prevent bacterial growth.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s13567-018-0517-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5828400PMC
February 2018

Development of an Incremental Sit-to-Stand Exercise for Aerobic Fitness Evaluation.

Int J Sports Med 2017 Oct 11;38(11):827-832. Epub 2017 Sep 11.

Physical Therapy, Shinshu University, School of Health Science, Matsumoto, Japan.

Incremental sit-to-stand exercise (ISTS) is an incremental exercise test using external signals to control the sit-to-stand rate in a given time frame. This study aimed to investigate the concurrent validity and reproducibility of the ISTS in aerobic fitness evaluation among healthy elderly women. Sixteen elderly women performed the ISTS and cycle ergometer test at 3-day to 2-week intervals, and six of the participants performed the ISTS twice. Peak oxygen uptake (VO), peak heart rate and completion time on the ISTS and cycle ergometer test were determined. Measured peak VO on the cycle ergometer test was significantly related to the peak VO (r=0.80, P<0.05) and the completion time (r=0.65, P<0.05) on the ISTS. The intraclass correlation coefficients were 0.96 for peak VO values and 0.91 for the completion time values during the two ISTSs. In conclusion, the ISTS is a valid, reproducible and safe test for aerobic fitness evaluation in healthy elderly women.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1055/s-0043-114008DOI Listing
October 2017

Exfoliation rate of mammary epithelial cells in milk on bovine mastitis caused by Staphylococcus aureus is associated with bacterial load.

Anim Sci J 2018 Jan 11;89(1):259-266. Epub 2017 Sep 11.

Dairy Hygiene Unit, Division of Pathology and Pathophysiology, Hokkaido Research Station, National Institute of Animal Health, NARO, Sapporo, Hokkaido, Japan.

The exfoliation rate of mammary epithelial cells (MECs) in milk is affected by physiological, breeding and environmental factors. Little is known about the relationship between the MEC exfoliation into milk and mammary-infected Staphylococcus aureus (S. aureus) load on bovine mastitis caused by S. aureus. The aim of this study was to investigate the relationship between S. aureus load and the proportion of MEC exfoliation in milk using five substantial bovine mastitis models. In 64 randomly extracted milk samples from udders at 3-21 days after S. aureus infusion, there were various samples with different numbers of S. aureus counts and somatic cell counts. No significant correlations were found between the S. aureus counts and somatic cell count (r = 0.338). In contrast, a significant correlation was noted between S. aureus counts and the proportion of cytokeratin-positive cells in the milk from the infused udders (r = 0.734, P < 0.01). In conclusion, the increasing MEC exfoliation rate in milk from mastitis udders caused by S. aureus may contribute to reduced milk yield.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/asj.12886DOI Listing
January 2018

Effect of intramammary infusion of recombinant bovine GM-CSF and IL-8 on CMT score, somatic cell count, and milk mononuclear cell populations in Holstein cows with Staphylococcus aureus subclinical mastitis.

Vet Res Commun 2017 Sep 9;41(3):175-182. Epub 2017 Mar 9.

Hokkaido Research Station, National Institute of Animal Health, NARO, 4 Hitsujigaoka, Toyohira, Sapporo, Hokkaido, 062-0045, Japan.

The effect of intramammary infusion of recombinant bovine granulocyte-macrophage colony-stimulating factor (rbGM-CSF) and interleukin-8 (rbIL-8) on mononuclear cell populations in quarters, somatic cell count (SCC) and the California Mastitis Test (CMT) score were investigated. From the selected cows with naturally occurring Staphylococcus aureus subclinical mastitis, one quarter of each cow were selected for the infusions of rbGM-CSF (400 μg/5 mL/quarter, n = 9), rbIL-8 (1 mg/5 mL/quarter, n = 9), and phosphate-buffered saline (5 mL/quarter, n = 7). The CMT score of both cytokines post infusion temporarily increased between days 0 and 1 and significantly decreased between days 7 and 14 compared to the preinfusion level. The SCC on day 14 after infusions of rbGM-CSF tended to be lower than that of the control group. The percentage of CD14+ cells increased on days 1 and 2 post infusion of rbGM-CSF. The percentage of CD4+ and CD8+ cells also increased on days 2 and 3, suggesting that the infusion of rbGM-CSF enhanced cellular immunity in the mammary gland. In contrast, the percentage of CD14+ cells decreased on days 0.25 and 1 post infusion of rbIL-8. No significant changes in the percentages of CD4+ and CD8+ cells in milk after infusion of rbIL-8 were evident during the experimental period, which suggested that rbIL-8 had little effect on the function of T cells in the mammary gland. These results indicated that rbGM-CSF and rbIL-8 decreased the CMT score by a different mechanism and may have a potential as therapeutic agents for subclinical mastitis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s11259-017-9684-yDOI Listing
September 2017

Effect of Different Seat Heights during an Incremental Sit-To-Stand Exercise Test on Peak Oxygen Uptake in Young, Healthy Women.

J Sports Sci Med 2016 Sep 5;15(3):410-416. Epub 2016 Aug 5.

School of Health Sciences, Department of Physical Therapy, Shinshu University , Matsumoto, Nagano, Japan.

'Sit-to-stand' exercise uses the repetitive motion of standing up and sitting down in a chair, a common activity of daily living. A new assessment using an incremental sit-to-stand exercise test employs an external sound to control the speed of standing-up and allows increases in work rate. The aims of the study were to examine the effect of different seat heights on peak oxygen uptake (peak VO) during an incremental sit-to-stand exercise and to assess any difference between peak VO values during incremental sit-to-stand exercise compared with a cycle ergometer test. Thirteen healthy young women (age: 23.1 ± 2.6 years, height: 1.61 ± 0.06 m, body mass: 51.9 ± 7.4 kg·m) participated in four incremental sit-to-stand tests with different seat heights and cycle tests in random order. The seat heights were adjusted to 100%, 80%, 120%, and 140% of knee height distance (100%, 80%, 120%, and 140% incremental sit-to-stand exercise, respectively). The peak VO and completion time were measured during incremental sit-to-stand and cycle ergometer tests, and repeated-measures analysis of variance and Student's paired t-test with Holm's method were used to evaluate differences between these variables. The peak VO values increased by about 10-12 mL·min·kg as the seat height on the ISTS decreased over a 60% range of lower leg lengths. The peak VO values on the 80%, 100%, 120%, and 140% incremental sit-to-stand tests were about 11%, 25%, 40%, and 50% lower than that on the cycle ergometer test, respectively. The peak VO on the incremental sit-to-stand test increased as seat height decreased. These findings are useful to determine which seat height on the incremental sit-to-stand tests test is suitable for different populations.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4974853PMC
September 2016

The cell wall component lipoteichoic acid of Staphylococcus aureus induces chemokine gene expression in bovine mammary epithelial cells.

J Vet Med Sci 2016 Oct 20;78(9):1505-1510. Epub 2016 May 20.

Hokkaido Research Station, National Institute of Animal Health, NARO, Sapporo, Hokkaido 062-0045, Japan.

Staphylococcus aureus (SA) is a major cause of bovine mastitis, but its pathogenic mechanism remains poorly understood. To evaluate the role of lipoteichoic acid (LTA) in the immune or inflammatory response of SA mastitis, we investigated the gene expression profile in bovine mammary epithelial cells stimulated with LTA alone or with formalin-killed SA (FKSA) using cap analysis of gene expression. Seven common differentially expressed genes related to immune or inflammatory mediators were up-regulated under both LTA and FKSA stimulations. Three of these genes encode chemokines (IL-8, CXCL6 and CCL2) functioning as chemoattractant molecules for neutrophils and macrophages. These results suggest that the initial inflammatory response of SA infection in mammary gland may be related with LTA induced chemokine genes.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5059380PMC
http://dx.doi.org/10.1292/jvms.15-0706DOI Listing
October 2016

Cyclophilin A is a new M cell marker of bovine intestinal epithelium.

Cell Tissue Res 2016 06 22;364(3):585-597. Epub 2016 Feb 22.

Cellar Biology Laboratory, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori Amamiyamachi, Aoba-ku, 981-8555, Sendai, Japan.

Microfold (M) cells in the follicle-associated epithelium (FAE) of Peyer's patches contribute to the mucosal immune response by the transcytosis of microorganisms. The mechanism by which M cells take up microorganisms, and the functional proteins by which they do this, are not clear. In order to explore one such protein, we developed a 2H5-F3 monoclonal antibody (2H5-F3 mAb) through its binding to bovine M cells, and identified the antibody reactive molecule as cyclophilin A (Cyp-A). The localization patterns of Cyp-A were very similar to the localization pattern of cytokeratin (CK) 18-positive M cells. Cyp-A was identified at the luminal surface of CK18-positive M cells in bovine jejunal and ileal FAE. The membranous localization of Cyp-A in the bovine intestinal cell line (BIE cells) increased as cells differentiated toward M cells, as determined by flow cytometry analysis. Additionally, BIE cells released Cyp-A to the extracellular space and the differentiation of BIE cells to M cells increased the secretion of Cyp-A, as determined by western blotting. Accordingly, Cyp-A may be localized in M cells in the small intestinal epithelium of cattle. The rise of the membranous localization and secretion of Cyp-A by differentiation toward M cells indicates that Cyp-A has an important role in the function of M cells. While Cyp-A of the M cell membrane may contribute to the uptake of viruses with peptidyl-prolyl cis-trans isomerase activity, in the extracellular space Cyp-A may work as a chemokine and contribute to the distribution of immuno-competent cells.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00441-015-2342-1DOI Listing
June 2016

Serotonin Improves High Fat Diet Induced Obesity in Mice.

PLoS One 2016 14;11(1):e0147143. Epub 2016 Jan 14.

Cellar Biology Laboratory, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori Amamiyamachi, Aoba-ku, Sendai, 981-8555, Japan.

There are two independent serotonin (5-HT) systems of organization: one in the central nervous system and the other in the periphery. 5-HT affects feeding behavior and obesity in the central nervous system. On the other hand, peripheral 5-HT also may play an important role in obesity, as it has been reported that 5-HT regulates glucose and lipid metabolism. Here we show that the intraperitoneal injection of 5-HT to mice inhibits weight gain, hyperglycemia and insulin resistance and completely prevented the enlargement of intra-abdominal adipocytes without having any effect on food intake when on a high fat diet, but not on a chow diet. 5-HT increased energy expenditure, O2 consumption and CO2 production. This novel metabolic effect of peripheral 5-HT is critically related to a shift in the profile of muscle fiber type from fast/glycolytic to slow/oxidative in soleus muscle. Additionally, 5-HT dramatically induced an increase in the mRNA expression of peroxisome proliferator-activated receptor coactivator 1α (PGC-1α)-b and PGC-1α-c in soleus muscle. The elevation of these gene mRNA expressions by 5-HT injection was inhibited by treatment with 5-HT receptor (5HTR) 2A or 7 antagonists. Our results demonstrate that peripheral 5-HT may play an important role in the relief of obesity and other metabolic disorders by accelerating energy consumption in skeletal muscle.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0147143PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4713156PMC
July 2016

Validity and Reproducibility of an Incremental Sit-To-Stand Exercise Test for Evaluating Anaerobic Threshold in Young, Healthy Individuals.

J Sports Sci Med 2015 Dec 24;14(4):708-15. Epub 2015 Nov 24.

Department of Rehabilitation, Matsumoto City Hospital , Matsumoto, Nagano, Japan.

Sit-to-stand exercise (STS) is a common activity of daily living. The objectives of the present study were: 1) to assess the validity of aerobic fitness measurements based on anaerobic thresholds (ATs), during incremental sit-to-stand exercise (ISTS) with and without arm support compared with an incremental cycle-ergometer (CE) test; and 2) to examine the reproducibility of the AT measured during the ISTSs. Twenty-six healthy individuals randomly performed the ISTS and CE test. Oxygen uptakes at the AT (AT-VO2) and heart rate at the AT (AT-HR) were determined during the ISTSs and CE test, and repeated-measures analyses of variance and Tukey's post-hoc test were used to evaluate the differences between these variables. Pearson correlation coefficients were used to assess the strength of the relationship between AT-VO2 and AT-HR during the ISTSs and CE test. Data analysis yielded the following correlations: AT-VO2 during the ISTS with arm support and the CE test, r = 0.77 (p < 0.05); AT-VO2 during the ISTS without arm support and the CE test, r = 0.70 (p < 0.05); AT-HR during the ISTS with arm support and the CE test, r = 0.80 (p < 0.05); and AT-HR during the ISTS without arm support and the CE test, r = 0.66 (p < 0.05). The AT-VO2 values during the ISTS with arm support (18.5 ± 1.9 mL·min(-1)·kg(-1)) and the CE test (18.4 ± 1.8 mL·min(-1)·kg(-1)) were significantly higher than those during the ISTS without arm support (16.6 ± 1.8 mL·min(-1)·kg(-1); p < 0.05). The AT-HR values during the ISTS with arm support (126 ± 10 bpm) and the CE test (126 ± 13 bpm) were significantly higher than those during the ISTS without arm support (119 ± 9 bpm; p < 0.05). The ISTS with arm support may provide a cardiopulmonary function load equivalent to the CE test; therefore, it is a potentially valid test for evaluating AT-VO2 and AT-HR in healthy, young adults. Key pointsThe ISTS is a simple test that varies only according to the frequency of standing up, and requires only a small space and a chair.The ISTS with arm support is valid and reproducible, and is a safe test for evaluating AT in healthy young adults.For evaluating the AT, the ISTS may serve as a valid alternative to conventional CPX, using either a cycle ergometer or treadmill, in cases where the latter methods are difficult to implement.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4657412PMC
December 2015

Effect of myostatin on chemokine expression in regenerating skeletal muscle cells.

Cells Tissues Organs 2013 3;198(1):66-74. Epub 2013 Jul 3.

Laboratory of Functional Morphology, Graduate School of Agricultural Science, Tohoku University, Sendai, Japan.

Transforming growth factor-β (TGF-β) is implicated in the regulatory expression of chemokines that control multiple steps in myogenesis. However, it remains to be established whether myostatin, a member of the TGF-β superfamily, affects chemokine expression in skeletal muscle. We investigated the effects of myostatin on the expression of mRNAs and proteins for 4 chemokines (CXCL1, CXCL2, CXCL6, CCL2) in intact and regenerating musculus longissimus thoracis from normal-muscled (NM) and double-muscled (DM) cattle. These chemokines were expressed in regenerating muscle, and their expression was always lower in DM than in NM cattle. Immunohistochemistry revealed that CXCL1 and CXCL6 were detected in the regenerating areas of myoblasts and myotubes in both NM and DM cattle. In cultures of myoblasts isolated from the regenerating muscles, significantly less CXCL1, CXCL2 and CCL2 mRNA was expressed in DM myoblasts than in NM myoblasts during the proliferating stage (P-stage). The expression of CXCL1, CXCL2 and CCL2 mRNAs in NM myoblasts and CXCL1, CXCL2 and CXCL6 mRNAs in DM myoblasts decreased upon switching from P-stage to fusion stage (F-stage). Also, the expression of CXCL1, CXCL2 and CXCL6 mRNAs was significantly lower in DM than in NM myoblasts during the F-stage. The addition of 100 ng/ml myostatin during the F-stage attenuated the expression of CXCL1 and CXCL2 mRNAs and augmented that of CCL2. These results show for the first time that myostatin regulates the differential expression of chemokines in skeletal muscle cells.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1159/000351462DOI Listing
February 2014

Cytokeratin 18 is a specific marker of bovine intestinal M cell.

Am J Physiol Gastrointest Liver Physiol 2011 Mar 30;300(3):G442-53. Epub 2010 Dec 30.

Cellular Biology Laboratory, Graduate School of Agricultural Science, Tohoku Univ., Sendai, Miyag, Japan.

Microfold (M) cells in the follicle-associated epithelium (FAE) of Peyer's patches have an important role in mucosal immune responses. A primary difficulty for investigations of bovine M cells is the lack of a specific molecular marker. To identify such a marker, we investigated the expression of several kinds of intermediate filament proteins using calf Peyer's patches. The expression patterns of cytokeratin (CK) 18 in jejunal and ileal FAE were very similar to the localization pattern of M cells recognized by scanning electron microscopy. Mirror sections revealed that jejunal CK18-positive cells had irregular and sparse microvilli, as well as pocket-like structures containing lymphocytes, typical morphological characteristic of M cells. However, CK18-negative cells had regular and dense microvilli on their surface, typical of the morphology of enterocytes. In contrast, CK20 immunoreactivity was detected in almost all villous epithelial cells and CK18-negative cells in the FAE. CK18-positive proliferating transit-amplifying cells in the crypt exchanged CK18 for CK20 above the mouth of the crypt and after moving to the villi; however, CK18-positive M cells in the crypt continued their expression of CK18 during movement to the FAE region. Terminal deoxynucleotidyl-transferase-mediated deoxyuridine-triphosphate-biotin nick-end labeling-positive apoptotic cells were specifically detected at the apical region of villi and FAE in the jejunum and ileum, and all were also stained for CK20. These data indicate that CK18 may be a molecular marker for bovine M cells in FAE and that M cells may transdifferentiate to CK20-positive enterocytes and die by apoptosis in the apex of the FAE.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1152/ajpgi.00345.2010DOI Listing
March 2011