Toa Pharmaceutical Co.

Tokyo, Japan

Toa Pharmaceutical Co.

Tokyo, Japan
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Jin J.S.,RIKEN | Touyama M.,RIKEN | Kibe R.,RIKEN | Tanaka Y.,RIKEN | And 12 more authors.
Beneficial Microbes | Year: 2013

The intestinal microbiota composition of 92 volunteers living in Japan was identified following the consumption of 'identical meals' (1,879 kcal/day) for 3 days. When faecal samples were analysed by terminal restriction fragment length polymorphism with several primer-restriction enzyme systems and then clustered, the patterns could be divided into 2 clusters. Contribution tests and partition modelling showed that OTU211 of the 35f-MspI system and OTU237 of the 35f-AluI system were key factors in the distribution of these groups. However, significant differences among these groups in terms of body mass index and age were not observed. ©2013 Wageningen Academic Publishers.


PubMed | Miyagi University, Biofermin Kobe Research Institute, Toa Pharmaceutical Co., Yakult Central Institute for Microbiological Research and 4 more.
Type: Journal Article | Journal: Bioscience of microbiota, food and health | Year: 2014

The intestinal microbiota compositions of 92 men living in Japan were identified following consumption of identical meals for 3 days. Fecal samples were analyzed by terminal restriction fragment length polymorphism with 4 primer-restriction enzyme systems, and the 120 obtained operational taxonomic units (OTUs) were analyzed by Data mining software focusing on the following 5 characteristics, namely, age, body mass index, present smoking habit, cessation period of previous smokers and drinking habit, according to the answers of the subjects. After performing Data mining analyses with each characteristic, the details of the constructed Decision trees precisely identified the subjects or discriminated them into various suitable groups. Through the pathways to reach the groups, practical roles of the related OTUs and their quantities were clearly recognized. Compared with the other identification methods for OTUs such as bicluster analyses, correlation coefficients and principal component analyses, the clear difference of this Data mining technique was that it set aside most OTUs and emphasized only some closely related ones. For example for a selected characteristic, such as smoking habit, only 7 OTUs out of 120 were able to identify all smokers, and the remaining 113 OTUs were thought of as data noise for smoking. Data mining analyses were affirmed as an effective method of subject discrimination for various physiological constitutions. The species of bacteria that were closely related to heavy smokers, i.e., HaeIII-291, were also discussed.


Huang A.,Hiroshima University | Shibata E.,Hiroshima University | Nishimura H.,Hiroshima University | Igarashi Y.,Toa Pharmaceutical Co. | And 2 more authors.
Journal of Poultry Science | Year: 2013

The aim of this study was to determine the effects of probiotics on T cell subsets induction in the intestine of broiler chicks. Day-old male broiler chicks were fed with or without probiotics consisting of Streptococcus faecalis, Clostridium buthricum, Bacillus mesentericus (probiotics group and control group, respectively). Cryostat sections of their ileum, cecum and rectum at day 0, 7 and 14 of feeding were immunostained for CD4, CD8 and TCRγδ, and the frequencies of positive cells in the mucosal tissue were analyzed. The CD4+, CD8+, TCRγδ+ T cells were localized in the lamina propria ofintestinal mucosa in all birds. At day 7 and 14, CD8+ T cells were localized also in the mucosal epithelium ofall segments in the probiotics group and ofcecum in control group, and TCR γδ+ T cells were observed in the mucosal epithelium ofall birds. The requencies ofCD4 +, CD8+, TCRγδ+ T cells were increased with age from day 0 to day 14 in both control and probiotics groups. The frequency of CD8+ T cells was significantly greater in probiotics group than control group in the ileum and rectum (P<0.05) and the cecum (P<0.01) at day 7. There were no significant differences in the frequency of CD4+ and TCRγδ+ T cells between control and probiotics groups in all intestinal segments at day 7 and 14. The ratio ofCD8 +/CD4+ T cells was greater than 1.0 in all tissues. The ratio in the ileum at day 7 was significantly greater in the probiotics group than control group (P< 0.05). These results further suggest that probiotics cause an influx of the CD8+ T cells into the intestinal mucosa, which may enhance the intestinal immunity by CD8+ T cells in young chicks. © 2013, Japan Poultry Science Association.


Mohammed E.S.I.,Hiroshima University | Igarashi Y.,Toa Pharmaceutical Co. | Isobe N.,Hiroshima University | Yoshimura Y.,Hiroshima University
Journal of Poultry Science | Year: 2015

The aim of this study was to determine the effects of probiotics-feeding on the gene expression and protein localization ofavian β-defensins (AvBDs) in the proventriculus of broiler chicks. Male broiler chicks were arranged in 3 groups: control group, probiotics group I and probiotics group II, which were fed with starter rations containing 0%, 0.2% or 0.4% probiotics, respectively, from day 0 (D0; at one day old) to D14. Proventriculi in all groups were collected at D0, D7 and D14 for analysis of AvBDs expression and AvBD12 protein localization. The expression of AvBDs genes was examined by reverse transcription-PCR and changes in the expression upon probiotics-feeding were examined by real-time PCR. The AvBD12 localization was examined by immunohistochemistry, and density of immunoreaction products was examined by image analysis under a microscope. Out of14 AvBDs genes, seven AvBDs were detected in the proventriculus ofchicks, namely, AvBD1, 2, 4, 6, 7, 10 and 12. The expression ofthe 7 detected genes did not show any significant differences between control and probiotics groups at D7 and D14. The immunoreactive (ir) -AvBD12 was localized in surface epithelium and cells in the connective tissues of proventricular glands. The ir-AvBD12 density in surface epithelium was significantly higher at D7 than at D0 or D14 in control group. At D7 and D14, the ir-AvBD12 density was significantly lower in probiotics groups than in control group. The ir-AvBD12 cells in proventricular gland increased in number with age; however, there were no significant differences between control and probiotics groups at D7 and D14. These results suggest that, although probioticsfeeding does not affect the gene expression of AvBDs, it may induce AvBD12 secretion from the surface epithelium of the proventriculus in broiler chicks. © 2015, Japan Poultry Science Association.


Shiozaki A.,University of Toyama | Yoneda S.,University of Toyama | Yoneda N.,University of Toyama | Yonezawa R.,University of Toyama | And 3 more authors.
PLoS ONE | Year: 2014

Preterm birth is a leading cause of perinatal morbidity and mortality. Studies using a cultivation method or molecular identification have shown that bacterial vaginosis is one of the risk factors for preterm birth. However, an association between preterm birth and intestinal microbiota has not been reported using molecular techniques, although the vaginal microbiota changes during pregnancy. Our aim here was to clarify the difference in intestinal and vaginal microbiota between women with preterm birth and women without preterm labor. 16S ribosomal ribonucleic acid genes were amplified from fecal and vaginal DNA by polymerase chain reaction. Using terminal restriction fragment length polymorphism (T-RFLP), we compared the levels of operational taxonomic units of both intestinal and vaginal flora among three groups: pregnant women who delivered term babies without preterm labor (non-PTL group) (n=20), those who had preterm labor but delivered term babies (PTL group) (n=11), and those who had preterm birth (PTB group) (n=10). Significantly low levels of Clostridium subcluster XVIII, Clostridium cluster IV, Clostridium subcluster XIVa, and Bacteroides, and a significantly high level of Lactobacillales were observed in the intestinal microbiota in the PTB group compared with those in the non-PTL group. The levels of Clostridium subcluster XVIII and Clostridium subcluster XIVa in the PTB group were significantly lower than those in the PTL group, and these levels in the PTL group were significantly lower than those in non-PTL group. However, there were no significant differences in vaginal microbiota among the three groups. Intestinal microbiota in the PTB group was found to differ from that in the non-PTL group using the T-RFLP method. © 2014 Shiozaki et al.


PubMed | Toa Pharmaceutical Co. and University of Toyama
Type: Journal Article | Journal: PloS one | Year: 2014

Preterm birth is a leading cause of perinatal morbidity and mortality. Studies using a cultivation method or molecular identification have shown that bacterial vaginosis is one of the risk factors for preterm birth. However, an association between preterm birth and intestinal microbiota has not been reported using molecular techniques, although the vaginal microbiota changes during pregnancy. Our aim here was to clarify the difference in intestinal and vaginal microbiota between women with preterm birth and women without preterm labor. 16S ribosomal ribonucleic acid genes were amplified from fecal and vaginal DNA by polymerase chain reaction. Using terminal restriction fragment length polymorphism (T-RFLP), we compared the levels of operational taxonomic units of both intestinal and vaginal flora among three groups: pregnant women who delivered term babies without preterm labor (non-PTL group) (n = 20), those who had preterm labor but delivered term babies (PTL group) (n = 11), and those who had preterm birth (PTB group) (n = 10). Significantly low levels of Clostridium subcluster XVIII, Clostridium cluster IV, Clostridium subcluster XIVa, and Bacteroides, and a significantly high level of Lactobacillales were observed in the intestinal microbiota in the PTB group compared with those in the non-PTL group. The levels of Clostridium subcluster XVIII and Clostridium subcluster XIVa in the PTB group were significantly lower than those in the PTL group, and these levels in the PTL group were significantly lower than those in non-PTL group. However, there were no significant differences in vaginal microbiota among the three groups. Intestinal microbiota in the PTB group was found to differ from that in the non-PTL group using the T-RFLP method.


PubMed | TOA Pharmaceutical Co.
Type: Evaluation Studies | Journal: Biological & pharmaceutical bulletin | Year: 2014

Strain-specific polymerase chain reaction (PCR) primers for detection of Bacillus mesentericus strain TO-A (BM TO-A) were developed. The randomly amplified polymorphic DNA (RAPD) technique was used to produce potential strain-specific markers. A 991-bp RAPD marker found to be strain-specific was sequenced, and two primer pairs specific to BM TO-A were constructed based on this sequence. In addition, we explored a more specific DNA region using inverse PCR, and designed a strain-specific primer set for use in real-time quantitative PCR (qPCR). These primer pairs were tested against 25 Bacillus subtilis strains and were found to be strain-specific. After examination of the detection limit and linearity of detection of BM TO-A in feces, the qPCR method and strain-specific primers were used to quantify BM TO-A in the feces of healthy volunteers who had ingested 310(8) colony forming unit (CFU) of BM TO-A per day in tablets. During the administration period, BM TO-A was detected in the feces of all 24 subjects, and the average number of BM TO-A detected using the culture method and qPCR was about 10(4.8) and 10(5.8) cells per gram of feces, respectively. Using the qPCR method, BM TO-A was detected in the feces of half of the subjects 3d after withdrawal, and was detected in the feces of only one subject 1 week after withdrawal. These results suggest that the qPCR method using BM TO-A strain-specific primers is useful for the quantitative detection of this strain in feces.

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