Yukinori Yoshimura

Hiroshima University, Hirosima, Hiroshima, Japan

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Publications (104)143.27 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: The aim of the present study was to examine the effect of the somatic cell count (SCC) in milk on reproductive performance, such as pregnancy status in the prepartum period and ovarian function in the postpartum period, in dairy cows. Blood samples were collected every week from one month prepartum to parturition in order to measure the concentrations of 13,14-dihydro-15-keto-PGF2α (PGFM), estrone sulfate (E1S) and progesterone. Milk samples were collected three times per week in both the prepartum (for one month before the dry period) and postpartum periods (for 3 months immediately after parturition) to measure the SCC. Progesterone was also determined in the whole milk of postpartum cows to define the day of the first ovulation. In the prepartum period, the maximum SCC negatively correlated with the pregnancy period (r = -0.77), but not the calf birth weight. Positive and negative correlations were observed between the average SCC and PGFM or progesterone concentrations in plasma, respectively (r = 0.84 or -0.92, respectively), at 39 weeks of pregnancy. In the postpartum period, a correlation was observed between the day of the first ovulation and both the average and maximum SCC (r = -0.74 and -0.75, respectively), whereas days open was not related to the SCC. These results suggest that a high SCC in the prepartum period may advance parturition by increasing PGF2α and decreasing progesterone and that the first ovulation in the postpartum period was delayed by a high SCC.
    Journal of Reproduction and Development 09/2014; · 1.76 Impact Factor
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    ABSTRACT: The aim of the present study was to examine changes in innate immune factors in the milk of mastitic dairy cows treated with antibiotics. Cows in the antibiotics group (n = 13) were infused into the mammary gland with cefazolin on the sixth day after mastitis was diagnosed (the day of the mastitis diagnosis = day −6). The control group (n = 12) was not treated. Milk samples were collected once every 2 days from days −6 to 12 and somatic cell count (SCC), lingual antimicrobial peptide (LAP), and lactoferrin (LF) concentrations and lactoperoxidase (LPO) activity were measured. SCC and LF concentrations in the antibiotics group markedly decreased after the antibiotic treatment. When cows in the antibiotics group were divided according to SCC on day 0, LAP concentrations and LPO activity in cows with a lower SCC on day 0 (<5 × 106 cell/mL) were significantly higher and lower than those in cows with a higher SCC, respectively. These results suggest that LF concentration decreased with decrease in SCC after treatment and that LAP concentration and LPO activity differed depending on the severity of mastitis. This is the first report to reveal the dynamics of innate immune factor in milk of cows treated for clinical mastitis.
    Animal Science Journal 09/2014; · 1.04 Impact Factor
  • B Ariyadi, N Isobe, Y Yoshimura
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    ABSTRACT: We previously reported that bacterial lipopolysaccharide (LPS), a ligand of Toll-like receptor 4 (TLR4), induced mucin mRNA to enhance the mucosal barrier in the hen vagina. The aim of this study was to determine the intracellular signaling molecules for that mucin induction, and the effect of molting and estrogen on their expression. The expression of TLR4, its adaptor molecules, and transcriptional factors in the vaginal mucosa of laying and molting hens treated with or without estradiol was examined by reverse-transcription PCR. The expression of mucin in the cultured mucosal tissue stimulated by LPS together with inhibitors of transcriptional factors was analyzed by quantitative reverse-transcription PCR. The expression of TLR4, its adaptor molecule, namely, myeloid differentiation factor 88 (MyD88) or Toll-interleukin 1 receptor domain-containing adaptor-inducing IFN-β (TRIF), and transcriptional factors, namely, cFos and cJun, declined in molting hens compared with that in laying hens, and were upregulated by estradiol. In vagina of laying hens, mucin expression was upregulated by LPS, whereas it was suppressed by inhibitors of transcriptional factors, namely, ALLN (an inhibitor of IκB proteolysis), BAY-117085 (an NFκB inhibitor), U0126 [a mitogen-activated protein kinase (MAPK) inhibitor], and transhinone IIA [an activated protein 1 (AP-1) inhibitor]. These results suggest that a MyD88-dependent pathway downstream of TLR4 and transcriptional factors of NFκB and AP-1 participate in the induction of mucin expression by LPS in the vaginal mucosa. These signaling functions may decline during molting owing to the decline in the level of circulating estrogen. Such mucin expression system may play a role in the mucosal barrier against infection in the vaginal mucosa.
    Poultry Science 03/2014; 93(3):673-9. · 1.52 Impact Factor
  • Gong-Wei Zhang, Song-Jia Lai, Yukinori Yoshimura, Naoki Isobe
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    ABSTRACT: Cathelicidins are a family of antimicrobial peptides found in neutrophils and the epithelium that have broad-spectrum activity against bacteria. This study aimed to investigative the mRNA expression of cathelicidins and protein localization of cathelicidin-2 in the goat mammary gland and its secretion into milk. The mRNA expression of cathelicidins was examined in different regions of the mammary gland by reverse transcription PCR. A cathelicidin-2 antibody was raised in rabbits by immunization with a synthetic cathelicidin-2 peptide consisting of 17 amino acids. The protein localization of cathelicidin-2 was investigated in the mammary gland by immunohistochemistry. Skim milk was collected before (0h) and 2, 4, 8, 12, and 24h after the intramammary infusion of lipopolysaccharide and saline, and the concentration of cathelicidin-2 was examined by an enzyme immunoassay. The mRNAs of cathelicidin-1, 2, 3, 6, and 7 were expressed in both the teat and deep region of the mammary gland from healthy and mastitic goats. Immunoreactive cathelicidin-2 was not localized in the epithelial cells of the teat skin, teat cistern, or mammary alveoli, whereas it was localized in polymorphonuclear cells in the mammary gland and those collected from the blood and milk. Cathelicidin-2 was detected in skim milk by Western blotting. The concentration of cathelicidin-2 in milk increased 4h after the intramammary infusion of lipopolysaccharide. These results suggest that cathelicidin-2 is expressed in polymorphonuclear cells and is secreted into milk in goat.
    Veterinary Microbiology 02/2014; · 3.13 Impact Factor
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    ABSTRACT: The ovary and oviduct of the hen are susceptible to various pathogenic microorganisms, and infections in these organs may not only contaminate the eggs, but also disorder the egg formation. The immune function of the ovary and oviduct is essential to protect these tissues from infection as well as for the production of hygienic eggs. This paper reviews recent studies on the host defence system in the reproductive organs with reference to their innate immune functions, with emphasis on the important role of Toll-like receptors and avian β-defensins in this defence system.
    Avian biology research 01/2014; 7(1). · 0.67 Impact Factor
  • Takahiro Nii, Naoki Isobe, Yukinori Yoshimura
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    ABSTRACT: The aim of this study was to determine the mechanism by which the avian infectious bronchitis virus (IBV) affects eggshell formation. Attenuated IBV (aIBV group) or vehicle (control group) was injected into the oviductal magnum lumen of White Leghorn laying hens. The changes in the expression of genes related to eggshell formation (collagen types I and V, and CaBP-D28 K), densities of cytotoxic cells (CD8+ cells and TCR-γδ+ T cells) as well as gene expression of molecules related to cytotoxic immunoreaction (B-NK, perforin, granzyme and IL-2) and proinflammatory cytokines (IL-1β, IL-6 and IFN-γ) were examined by quantitative RT-PCR or immunohistochemistry in the isthmus and uterus. Gene expression of IL-1β and IL-6 receptors in the tubular gland cells of the isthmus and uterus was analyzed by RT-PCR. Gene expression of collagen type I, but not collagen type V, in the isthmus and CaBP-D28 K in the uterus was decreased in the aIBV group compared with that in the control. The frequencies of CD8+ cells and TCR-γδ+ T cells in the isthmus and uterus were significantly higher in the aIBV group than in the control group. The expression of cytotoxic molecular and proinflammatory cytokines was also higher in the aIBV group than in the control. The expression of IL-6 receptor, but not IL-1β receptor, was identified in the tubular gland cells in the isthmus and uterus. These results suggest that IBV infection causes disorder of eggshell formation by disturbing gene expression of collagen type I in the isthmus and CaBP-D28 K in the uterus, probably via the effects of substances from cytotoxic cells and proinflammatory cytokines.
    Theriogenology 01/2014; · 2.08 Impact Factor
  • G.W. Zhang, S.J. Lai, Y. Yoshimura, N. Isobe
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    ABSTRACT: Psoriasin (S100A7) is a member of the S100 protein family of calcium-binding proteins and plays a crucial role in local host defences. The present study aimed to identify the expression of S100A7 in the goat mammary gland and in milk. The goat S100A7 coding DNA sequence was identified using direct sequencing. An S100A7 antibody was raised in rabbits by immunization with a synthetic S100A7 peptide consisting of 13 amino acids. Messenger RNA expression and protein localization in different regions of a healthy mammary gland were detected by reverse transcription-polymerase chain reaction and immunohistochemistry. Changes in the concentration of S100A7 in the milk after an intramammary infusion of Escherichia coli lipopolysaccharide (LPS) were examined by an enzyme immunoassay. The goat S100A7 peptide had 98% and 86% sequence similarity to that of sheep and bovines, respectively. The S100A7 mRNA expression was higher in the teat and udder skin than in the cistern and parenchyma of the mammary gland. Immunoreactive S100A7 was localized in the epithelial cells of the alveolus and gland cistern, and stratified squamous epithelium of the teat. Psoriasin as a secreted protein was detectable in healthy milk, and an intramammary LPS infusion increased the concentration of S100A7 in the milk. The results suggest that S100A7 is produced in the epithelial cells of the mammary gland and is secreted into the milk.
    The Veterinary Journal. 01/2014;
  • B Ariyadi, N Isobe, Y Yoshimura
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    ABSTRACT: Mucins play an essential role as mucosal barrier to prevent invasion of pathogens in the oviductal tissue of hens. The aim of this study was to determine the effect of estradiol and lipopolysaccharide (LPS) on the mucin expression in the lower oviductal segments (vagina and uterus) of hens. The mucosal tissues of the vagina and uterus were collected from White Leghorn laying and molting hens, and molting hens with or without intramuscular injection with 1 mg of estradiol-benzoate (EB) daily for 7 d. Part of these tissues was cultured in TCM-199 culture medium with or without LPS (10, 100, or 1,000 ng/mL) for 1.5 or 3 h. Mucin expression in the mucosa of laying, molting, and EB-treated molting hens (EB-group) and in those tissues cultured with or without LPS was analyzed by quantitative reverse-transcription PCR. Cultured tissues were also processed for paraffin sections and stained with Alcian blue (AB). In both the vagina and uterus, mucin expression and density of AB-positive mucopolysaccharide were reduced in molting hens compared with laying hens, and upregulated by EB. Mucin expression in the cultured vagina and uterus tissues of laying and molting hens was upregulated by LPS in a dose- and time-dependent manner. However, there was no response to LPS for induction of mucin in the tissues of EB-group hens. The mucin expression level in the vagina and uterus tissues stimulated by LPS was lower in the EB-group hens than in laying and molting hens, and that in the uterus was lower in the molting hens than in laying hens. These results suggest that mucin expression is stimulated by LPS in the vagina and uterus of laying and molting hens. Estrogen may upregulate mucin expression in those tissues in association with epithelial development, whereas it may suppress the response to LPS for mucin induction. The mucin expression caused by LPS may enhance mucosal barrier function and play a role in preventing infections by bacteria in the vagina and uterus.
    Poultry Science 12/2013; 92(12):3205-13. · 1.52 Impact Factor
  • Naoki Isobe, Ayumi Shibata, Hirokazu Kubota, Yukinori Yoshimura
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    ABSTRACT: The present study was undertaken to examine whether potential levels of innate immune factors (lingual antimicrobial peptide (LAP), lactoferrin (LF) and lactoperoxidase (LPO)) in colostrum are associated with subsequent milk somatic cell count (SCC) in dairy cows. Quarter milk samples were collected daily for 1 week postpartum to measure LAP and LF concentrations and LPO activity. SCC in milk was determined weekly for 2 months postpartum and its correlations to concentrations of LAP and LF and LPO activity were examined. Only small variations of all immune factors were found among four udders in each individual cow, whereas there were great differences in these factors among cows. Negative correlation was detected only between LPO activity and mean and maximum SCC, whereas its relationship was not significant. LAP and LF concentrations were significantly correlated positively to mean, maximum and minimum SCC. These results suggest that the great difference in innate immune factors among animals and high LAP and LF concentrations in colostrum may be associated with subsequent high incidence of SCC increase.
    Animal Science Journal 09/2013; · 1.04 Impact Factor
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    ABSTRACT: Dectin-1 plays a critical role in the pathogenesis of intestinal inflammation by recognizing the pathogenic agents and mediating cytokine responses. The objective of this study was to establish the association between dectin-1 polymorphisms and susceptibility to non-specific digestive disorders (NSDD) and cytokine expression in rabbits. A total of seven coding SNPs were detected in dectin-1 gene. The genetic association between SNP (ss707197675A>G) and susceptibility to NSDD was evaluated using a case-control study (178 cases and 174 controls). The results revealed that the A allele was associated with an increased risk of developing NSDD in rabbits. The AA genotype significantly increased the genetic susceptibility to NSDD with odds ratio of 4.76 (95% confidence interval, 1.92-12.50, P = 0.0002) compared with GG and GA genotypes. We also experimentally induced NSDD in another independent growing rabbit population by feeding a low-fiber diet and subsequently investigated the cytokine mRNA expression. Among the four studied cytokines, the expression levels of IFN-γ, IL-17F, and IL-22 were increased 2.8 to 6.0 fold in AA genotype compared with GG genotype (P < 0.01). The greater IL-17F and IL-22 mRNA expressions indicated a positive correlation with severe intestinal inflammation (P < 0.05). The decreased expression of IL-10 was associated with severe intestinal inflammation (P = 0.006), but IL-10 expression was not influenced by dectin-1 genotype. In conclusion, polymorphism ss707197675 of dectin-1 is related with susceptibility to NSDD and increased expression of proinflammatory cytokines, and dectin-1 could be an important candidate gene associated with NSDD in rabbits.
    Journal of Animal Science 07/2013; · 2.09 Impact Factor
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    ABSTRACT: Immune function in the vagina of hen oviduct is essential to prevent infection by microorganisms colonizing in the cloaca. The aim of this study was to determine whether CpG oligodeoxynucleotide (CpG-ODN) stimulate the expression of avian β-defensins (AvBDs) in hen vaginal cells. Specific questions were whether CpG-ODN affects the expression of AvBDs and proinflammatory cytokines, and whether the cytokines affect AvBDs expression in vaginal cells. The dispersed vaginal cells of White Leghorn laying hens were cultured and stimulated by different doses of lipopolysaccharide (LPS), CpG-ODN, interleukin (IL)1β, or IL6. The cultured cell population contained epithelial cells, fibroblast-like cells and CD45-positive leukocytes. The immunoreactive AvBD 3, 10 and 12 were localized in the mucosal epithelium in the section of the vagina. The expression of AvBDs, IL1β, and IL6 was analyzed by quantitative RT-PCR. RT-PCR analysis showed the expression of AvBD 1, 3, 4, 5, 10, and 12 in the cultured vaginal cells without stimulation. Toll-like receptors (TLRs) 4 and 21, which recognize LPS and CpG-ODN, respectively, and IL1 and IL6 receptors (IL1R and IL6R) were also expressed in them. The expression of IL1β, IL6, and AvBD 10 and 12 was upregulated by LPS, whereas only IL1β and IL6 were upregulated by CpG-ODN. IL1β stimulation upregulated AvBD 1 and 3 expression, whereas IL6 stimulation did not cause changes in AvBDs expression. These results suggest that CpG-ODN derived from microbes upregulates the expression of IL1β and IL6 by interaction with TLR 21, and then IL1β induces AvBD 1 and 3 to prevent infection in the vagina.
    Reproduction 04/2013; · 3.56 Impact Factor
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    ABSTRACT: As stage progresses in the cystic follicle, granulosa cells are lost. We hypothesized that the granulosa and theca interna layers are detached in association with weakened expression of cell adhesion molecules such as cadherin (cell-cell adhesion) and integrin (cell-extracellular matrix adhesion) in cystic follicles. To elucidate this hypothesis, we immunolocalized these molecules in the granulosa and theca interna and compared them between cystic and small healthy follicles. Sections were immunostained with cadherin and integrin β1 antibodies and their localizations were compared. Cadherin-positive reaction was seen in the cytoplasma of all granulosa cells. No increase in the frequency of cadherin-positive area in the granulosa layers and the intensity of cadherin immunoreaction in the theca interna was detected in cystic follicles compared with healthy ones. A dense immunoreaction product of integrin β1 was detected in the theca interna in both cystic and healthy follicles. Intensity of integrin β1 -immuno reaction in the granulosa layers and integrin β1 -positive area in the theca interna was significantly lower in the cystic follicle than in the healthy follicles. These results suggest that granulosa and theca interna cells are detached while maintaining the cell-cell adhesion, resulting in the consequent loss of these layers from the cystic follicle.
    Animal Science Journal 04/2013; 84(4):303-309. · 1.04 Impact Factor
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    ABSTRACT: Lingual antimicrobial peptide (LAP) belongs to the ß-defensin family in cattle and is found in milk. LAP concentrations increase in milk from mastitic udders; however, the relationship between LAP concentrations and the somatic cell count (SCC) in milk remains to be elucidated in detail. Therefore, the present study was undertaken to investigate the relationship between LAP concentrations and the SCC in bovine milk to assess whether LAP may be used as an indicator of SCC. Milk was collected from 66 udders showing various SCCs. The SCC and LAP concentrations were measured in the milk. A significantly higher LAP concentration was observed in milk having 500-5000×10(3)cells/ml and >5000×10(3)cells/ml SCC groups than in lower SCC groups (<50×10(3)cells/ml and 50-500×10(3)cells/ml). A significantly positive correlation between LAP concentrations and SCCs in milk was observed (r=0.68). In milk samples with >26nM of LAP, 92.0% of milk samples had high SCCs (>200×10(3)cells/ml). The concentration of LAP in milk infected with Staphylococcus aureus, Streptococcus bovis, Streptococcus dysgalactiae, and Escherichia coli was significantly higher than that in uninfected milk. These results suggest that the concentration of LAP can be a useful indicator of the SCC in dairy cows.
    Veterinary Immunology and Immunopathology 03/2013; · 1.88 Impact Factor
  • Bambang Ariyadi, Naoki Isobe, Yukinori Yoshimura
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    ABSTRACT: Tight junctions in the mucosal epithelium have essential roles as a mucosal barrier to prevent invasion of microbes into the hen oviduct tissue. The aim of this study was to determine the effects of the egg-laying phase and estradiol on the expression of tight junction molecule "claudins" in the lower oviductal segments in hens. White Leghorn laying and molting hens were used. Molting hens were given either sesame oil (vehicle) or estradiol benzoate (N = 5 per group) via injection. The lower segments of oviduct (isthmus, uterus, and vagina) of these birds were collected. Gene expression of claudin-1, -3, -5, lipopolysaccharide-induced TNFα factor (LITAF), and IFNˠ was analyzed by quantitative reverse transcription polymerase chain reaction, and localization of claudin-1 was examined by immunohistochemistry. Permeability in the mucosal epithelium was assessed by intrauterine injection of fluorescein isothiocyanate-dextran. Expression of claudin-1, -3, and -5 genes and density of claudin-1 protein in the lower oviductal segments were higher in laying hens than in molting hens (P < 0.01); their expression was upregulated by estradiol (P < 0.01). Expression of LITAF and IFNˠ genes was higher in molting hens than in laying hens. More fluorescein isothiocyanate-dextran infiltrated into the intercellular space of the uterus mucosal epithelium in molting hens than in laying hens and estradiol-treated molting hens. In conclusion, we inferred that barrier functions of the mucosal epithelium in the lower oviductal segments might be disrupted because of reduced claudin expression in molting hens, which might increase the susceptibility of mucosal tissue during the molting phase.
    Theriogenology 11/2012; · 2.08 Impact Factor
  • M Abdelsalam, N Isobe, Y Yoshimura
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    ABSTRACT: The aim of this study was to determine the mechanism by which expression of avian β-defensins (AvBD) in the follicular theca tissue was regulated. It was examined whether their expression was stimulated directly by LPS or indirectly through proinflammatory cytokines (IL-1β and IL-6) induced by LPS. Theca tissues of ovarian follicles were collected from White Leghorn hens. The specimens of those theca tissues were cultured in TCM-199 culture medium and stimulated by lipopolysaccharide from Salmonella minnesota (LPS), recombinant chicken IL-1β, or recombinant chicken IL-6. In the first experiment, changes in the expression of IL-1β, IL-6, AvBD10, and AvBD12 in response to LPS stimulation were examined by quantitative reverse-transcription PCR. The AvBD10 and 12 had been known to be expressed in the theca. In the second experiment, changes in the expression of AvBD10 and 12 in response to recombinant chicken IL-1β or IL-6 stimulation were examined by quantitative reverse-transcription PCR. Density of AvBD12 protein after IL-1β stimulation that showed changes in the gene expression was analyzed by Western blotting. In the first experiment, LPS was able to induce IL-1β and IL-6, but not AvBD10 or AvBD12. In the second experiment, IL-1β was able to upregulate significantly the expression of AvBD12 mRNA and protein. However, IL-6 did not exert significant effects on the expression of AvBD10 and AvBD12. It is suggested that LPS may stimulate theca cells to produce proinflammatory cytokines, whereas, in turn, IL-1β stimulates those cells to synthesize AvBD12, which may be able to attack infectious gram-negative bacteria.
    Poultry Science 11/2012; 91(11):2877-84. · 1.52 Impact Factor
  • M Zhang, T Nii, N Isobe, Y Yoshimura
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    ABSTRACT: The aim of this study was to determine the expression profiles of Toll-like receptors (TLR) in the testis and epididymis of rooster and whether the expression of IL-1β, IL-6, CXCLi2, and TLR-4 was affected by lipopolysaccharide (LPS), a TLR-4 ligand. Roosters were intravenously injected with LPS or phosphate-buffered saline. Testes and epididymis were collected before and after 3 or 6 h postinjection. Total RNA was isolated from those tissues and expression of TLR and proinflammatory cytokines was analyzed by reverse-transcription PCR and quantitative real-time PCR. Reverse-transcription PCR analysis revealed that 7 of the known 10 chicken TLR in the testis and 9 of 10 in the epididymis were expressed. Expression of TLR-4 was found in both tissues. Expression of TLR-4 was significantly upregulated by LPS in the testis but not in the epididymis. Injection with LPS upregulated the expression of IL-1β, IL-6, and CXCLi2 in the testis and epididymis by 3 to 6 h postinjection. However, injection with phosphate-buffered saline (control) did not affect their expression. These results suggest that proinflammatory cytokines and chemokine expression was upregulated by LPS probably through TLR-4 activation, and thus the reproductive tissues are comprehensively equipped to deal with a pathogenic insult.
    Poultry Science 08/2012; 91(8):1997-2003. · 1.52 Impact Factor
  • B Ariyadi, N Isobe, Y Yoshimura
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    ABSTRACT: The aim of this study was to determine the differences in the mucin expression that forms a mucosal surface barrier in the oviduct between laying and molting hens. The lower segments of oviducts (isthmus, uterus, and vagina) of White Leghorn laying and molting hens were collected. Localization and gene expression of mucosal mucin were analyzed by quantitative reverse-transcription PCR of mucin mRNA and mucin5AC immunohistochemistry. Sugar residues were localized by lectin (WGA or Jacalin) histochemistry. Expression of mucin mRNA was significantly declined in the lower oviductal segments in molting compared with laying hens. Immunoreactive-mucin5AC was localized in the mucosal epithelium and on the epithelial surface of laying hens, whereas it was reduced in molting hens. Substances positively stained by WGA and Jacalin were identified on the surface of the mucosal epithelium in the lower oviductal segments in laying and molting hens. These results suggest that mucin synthesis in the lower segments of the oviduct is reduced, although the existence of WGA- and Jacalin-positive sugars may be kept even in the molting phase. The reduction of mucin synthesis may result in a decline of mucosal barrier function in the molting phase.
    Poultry Science 05/2012; 91(5):1173-8. · 1.52 Impact Factor
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    ABSTRACT: Lingual antimicrobial peptide (LAP), one of the β-defensins in bovines, and lactoferrin (LF) are synthesized in mammary epithelium and have bactericidal and bacteriostatic functions. However, it is not known whether they have similar expression patterns. Therefore, the present study was undertaken to compare (1) immunolocalization of LAP and LF in the mammary gland and (2) changes in concentration of these two components in milk after lipopolysaccharide (LPS) challenge. Bovine mammary tissues without LPS challenge were collected and their sections were immunostained with antibodies to LAP or LF. Milk from our previous study was collected every hour up to 12h and twice daily from d 1 to 7 after LPS challenge (the day of infusion was considered as d 0). These milk samples were measured for LAP but not LF in our previous report. Therefore, concentration of LF was measured by enzyme immunoassay in the present study. Epithelial cells of some alveoli showed immunopositive reaction for LF, but negative for LAP. Conversely, some alveoli were LAP positive in their epithelial cells but LF negative. Many alveoli had immunoreactions for neither LAP nor LF. The concentration of LAP in milk was elevated significantly at 3h after LPS infusion compared with pre-infusion values and remained at a high level until 12h. However, LF concentration in milk remained low at d 0 and increased at d 2. These results suggest that LAP and LF were mostly differentially localized in the alveolar epithelium in mammary glands. The different spatial expressions between them may be associated with their different temporal expression mechanisms.
    Veterinary Immunology and Immunopathology 11/2011; 145(1-2):499-504. · 1.88 Impact Factor
  • T Nii, Y Sonoda, N Isobe, Y Yoshimura
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    ABSTRACT: The goal of this study was to examine whether lipopolysaccharide (LPS) induces the expression of proinflammatory cytokines and chemokines and recruits T cells in the lower part of the oviduct, and whether that response to LPS is different between the laying and molting phase. White Leghorn laying and molting hens were intravenously injected with saline (control) or LPS. The uterus and vagina of oviducts were collected 3 or 6 h after injection, and used for reverse transcription PCR analysis of IL-1β, IL-6, IL-8 (CXCLi2), and lymphotactin (Lptn), and for immunohistochemical analysis for the frequency of CD4+ and CD8+ T cells. The expressions of IL-1β, IL-6, and CXCLi2 in the uterus and that of IL-1β in the vagina were upregulated in response to LPS 3 or 6 h after injection in both laying and molting hens. The CXCLi2 expression in the vagina was upregulated by LPS in laying hens, whereas those effects of LPS were not significant in molting hens. Expression of Lptn showed a tendency to be downregulated after 3 h, with recovery by 6 h after LPS injection. The frequency of CD4+ T cells tended to increase in response to LPS after 6 h in the lamina propria of the uterus and vagina in both laying and molting hens. The CD8+ T cell frequencies in the lamina propria of the uterus and vagina of laying hens increased in response to LPS after 6 h. However, in the molting hens, LPS stimulation resulted in CD8+ T cell increase in the vagina only and not in the uterus. These results suggest that expressions of proinflammatory cytokines and CXCLi2 chemokine are upregulated in association with T cell recruitment in response to LPS in the lower part of the oviduct, although CD8+ T cells in the uterus may be depressed during the molting phase. These immunoresponses may play roles in the defense against infection of the oviduct.
    Poultry Science 10/2011; 90(10):2332-41. · 1.52 Impact Factor
  • M Abdelsalam, N Isobe, Y Yoshimura
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    ABSTRACT: The aim of this study was to determine whether the expression of proinflammatory cytokines and chemokines in ovarian cells was changed to recruit heterophils and T cells in response to lipopolysaccharide (LPS), a gram-negative bacterial component. White Leghorn laying hens were intravenously injected with LPS or saline and their ovarian follicles and stroma were collected. Changes in the mRNA expression of IL-1β, IL-6, and CXCLi2 in the theca and granulosa layers and ovarian stroma were analyzed by quantitative reverse transcriptase PCR, whereas proteins of IL-1β and IL-6 were also identified by Western blot analysis. Localization of heterophil-like cells and CD4(+) and CD8(+) T cells was examined by general histology and immunohistochemistry. The expressions of IL-1β, IL-6, and CXCLi2 were significantly increased in the granulosa layer, theca layer, and stroma tissues by 3 to 6 h after LPS injection. Increase of IL-1β and IL-6 proteins in those tissues was also identified 12 h after LPS injection. The LPS stimulation resulted in the increased influx of heterophil-like cells and CD4(+) T cells, but not CD8(+) cells, in the theca layers of follicles. Saline injection affected neither expression of examined proinflammatory cytokines and chemokines nor frequencies of immunocompetent cells. These results suggest that ovarian follicular cells and stromal cells have the ability to express proinflammatory cytokines and chemokines, and their expression is upregulated by LPS in association with the recruitment of heterophil-like cells and T cells. These responses may play roles in local host defense in ovarian follicles.
    Poultry Science 09/2011; 90(9):2054-62. · 1.52 Impact Factor

Publication Stats

596 Citations
143.27 Total Impact Points

Institutions

  • 1997–2014
    • Hiroshima University
      • • Graduate School of Biosphere Sciences
      • • Faculty of Applied Biological Science
      • • Graduate School for International Development and Cooperation
      Hirosima, Hiroshima, Japan
  • 2013
    • Azabu University
      • School of Veterinary Medicine
      Sagamihara, Kanagawa-ken, Japan
  • 2009–2011
    • Bangladesh Agricultural University
      • Department of Poultry Science
      Mymensingh, Bangladesh