Publications by authors named "Gaku Harata"

15 Publications

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Dysbiosis of Ocular Surface Microbiota in Patients With Refractive Allergic Conjunctival Diseases.

Cornea 2021 Dec 8. Epub 2021 Dec 8.

Division of Ophthalmology, Department of Visual Sciences, Nihon University School of Medicine, Tokyo, Japan; and Technical Research Laboratory, Takanashi Milk Products Co., Ltd., Kanagawa, Japan.

Purpose: We investigated ocular surface microbiota dysbiosis in patients with refractory allergic conjunctival diseases (ACDs; stratified into mild and severe groups) treated with topical tacrolimus.

Methods: Patients (n = 21) with refractory ACDs (including vernal and atopic keratoconjunctivitis) actively treated with topical tacrolimus and 6 healthy controls were evaluated. Based on clinical scores and expression of specific cytokines on the ocular surface, patients with ACDs were divided into mild and severe groups using cluster analysis. The microbial composition of tear specimens collected from patients with mild and severe ACD and control subjects using the Schirmer test paper was determined through next-generation 16S rRNA sequencing analysis.

Results: Compared with healthy controls, patients with ACDs exhibited significantly decreased ocular surface microbiota α-diversity. Ocular surface microbiota mainly comprised members of the phyla Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria in all groups. The relative abundance of ocular surface microbiota in patients with ACDs was increased for phylum Firmicutes and decreased for phylum Proteobacteria (compared with control subjects). The genera Blautia (vs. mild ACD group) and Morganella (vs. control group) exhibited significantly increased abundance only in the severe ACD group.

Conclusions: The ocular surface microbiota in patients with severe ACD exhibited decreased diversity and exacerbation of dysbiosis compared with that in patients with mild ACD and control subjects. Patients with mild refractory ACD also exhibited decreased diversity of these microbiota. These alterations in microbiota indicated a change in the ocular surface of patients with refractory ACD (be it because of disease pathogenesis or topical immunomodulatory treatment).
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http://dx.doi.org/10.1097/ICO.0000000000002940DOI Listing
December 2021

Species- and Age/Generation-Dependent Adherence of to Human Intestinal Mucus In Vitro.

Microorganisms 2021 Mar 5;9(3). Epub 2021 Mar 5.

Department of Food, Aroma and Cosmetic Chemistry, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri, Hokkaido 099-2493, Japan.

Adhesion to intestinal mucus is the first event in the process by which intestinal microbes colonize the intestine. It plays a critical role in the initiation of interactions between gut microbes and host animals. Despite the importance, the adhesion properties of probiotics are generally characterized using porcine mucin; adhesion to human mucus has been poorly characterized. In the present study, human intestinal mucus samples were isolated from 114 fecal samples collected from healthy infants and adults. In initial screening, four out of the 13 beneficial microbes tested, including the type strain of TMC3115, GG, and subsp. Bb12, showed strong adhesion abilities to human mucus. The type strain of and TMC3115 adhered more strongly to neonatal and infant mucus than to adult mucus, while GG and Bb12 adhered more strongly to adult mucus than to infant mucus. Similar results were obtained for ten additional strains of . In conclusion, age/generation-related differences were observed in the adhesion properties of and other strains. A deeper symbiotic relationship may exist between infants, particularly neonates, and based on its enhanced adhesion to neonatal intestinal mucus.
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http://dx.doi.org/10.3390/microorganisms9030542DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7998455PMC
March 2021

Selection of Probiotics for Microbiota Modulation in Normal-Weight and Severely Obese Individuals: Focus on Gas Production and Interaction With Intestinal Epithelial Cells.

Front Microbiol 2021 9;12:630572. Epub 2021 Feb 9.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias, Consejo Superior de Investigaciones Científicas (IPLA-CSIC), Villaviciosa, Spain.

The intestinal microbiota plays important roles in the maintenance of health. Strategies aiming at its modulation, such as probiotics, have received a deal of attention. Several strains have been studied in different models; however, the correlation of results obtained with the data has been limited. This questions the usefulness of such selection models, traditionally relying on over-simplified tests, not considering the influence of the accompanying microbiota or focusing on microbiota composition without considering functional traits. Here we assess the potential of six , and strains in an model to determine their impact on the microbiota not just in terms of composition but also of functionality. Moreover, we compared the responses obtained in two different population groups: normal-weight and severely obese subjects. Fecal cultures were conducted to evaluate the impact of the strains on specific intestinal microbial groups, on the production of short-chain fatty acids, and on two functional responses: the production of gas and the interaction with human intestinal epithelial cells. The response to the different probiotics differed between both human groups. The addition of the probiotic strains did not induce major changes on the microbiota composition, with significant increases detected almost exclusively for the species added. Higher levels of gas production were observed in cultures from normal-weight subjects than in the obese population, with some strains being able to significantly reduce gas production in the latter group. Moreover, in obese subjects all the strains tested and GG were able to modify the response of the intestinal cells, restoring values similar to those obtained with the microbiotas of normal-weight subjects. Our results underline the need for the screening and selection of probiotics in a target-population specific manner by using appropriate models before enrolling in clinical intervention trials.
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http://dx.doi.org/10.3389/fmicb.2021.630572DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7899977PMC
February 2021

Effects of dietary fiber on vascular calcification by repetitive diet-induced fluctuations in plasma phosphorus in early-stage chronic kidney disease rats.

J Clin Biochem Nutr 2020 Nov 6;67(3):283-289. Epub 2020 Aug 6.

Graduate School of Human Science and Environment, University of Hyogo, 1-1-12 Shinzaike-Honcho, Himeji, Hyogo 670-0092, Japan.

Vascular calcification progresses under hyperphosphatemia, and represents a risk factor for cardiovascular disease in chronic kidney disease (CKD) patients. We recently indicated that phosphorus (P) fluctuations also exacerbated vascular calcification in early-stage CKD rats. Dietary fiber intake is reportedly associated with cardiovascular risk. This study investigated the effects of dietary fiber on vascular calcification by repeated P fluctuations in early-stage CKD rats. Unilateral nephrectomy rats were used as an early-stage CKD model. For 36 days, a P fluctuation (LH) group was fed low-P (0.02% P) and high-P (1.2% P) diets alternating every 2 days, and a P fluctuation with dietary fiber intake (LH + F) group was fed low-P and high-P diets containing dietary fiber alternating every 2 days. The effect on vascular calcification was measured calcium content. Effects on uremic toxin were measured levels of indoxyl sulfate (IS) and investigated gut microbiota. The LH + F group showed significantly reduced vessel calcium content compared to the LH group. Further, dietary fiber inhibited increases in blood levels of IS after intake of high-P diet, and decreased uremic toxin-producing intestinal bacteria. Dietary fiber may help suppress progression of vascular calcification due to repeated P fluctuations in early-stage CKD rats by decreasing uremic toxin-producing intestinal bacteria.
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http://dx.doi.org/10.3164/jcbn.20-46DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7705083PMC
November 2020

Sharing of human milk oligosaccharides degradants within bifidobacterial communities in faecal cultures supplemented with Bifidobacterium bifidum.

Sci Rep 2018 09 18;8(1):13958. Epub 2018 Sep 18.

Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan.

Gut microbiota of breast-fed infants are generally rich in bifidobacteria. Recent studies show that infant gut-associated bifidobacteria can assimilate human milk oligosaccharides (HMOs) specifically among the gut microbes. Nonetheless, little is known about how bifidobacterial-rich communities are shaped in the gut. Interestingly, HMOs assimilation ability is not related to the dominance of each species. Bifidobacterium longum susbp. longum and Bifidobacterium breve are commonly found as the dominant species in infant stools; however, they show limited HMOs assimilation ability in vitro. In contrast, avid in vitro HMOs consumers, Bifidobacterium bifidum and Bifidobacterium longum subsp. infantis, are less abundant in infant stools. In this study, we observed altruistic behaviour by B. bifidum when incubated in HMOs-containing faecal cultures. Four B. bifidum strains, all of which contained complete sets of HMO-degrading genes, commonly left HMOs degradants unconsumed during in vitro growth. These strains stimulated the growth of other Bifidobacterium species when added to faecal cultures supplemented with HMOs, thereby increasing the prevalence of bifidobacteria in faecal communities. Enhanced HMOs consumption by B. bifidum-supplemented cultures was also observed. We also determined the complete genome sequences of B. bifidum strains JCM7004 and TMC3115. Our results suggest B. bifidum-mediated cross-feeding of HMOs degradants within bifidobacterial communities.
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http://dx.doi.org/10.1038/s41598-018-32080-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6143587PMC
September 2018

[PROBIOTICS AND ALLERGY].

Arerugi 2018;67(3):197-201

Takanashi Milk Products Co., Ltd. Technical Research.

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http://dx.doi.org/10.15036/arerugi.67.197DOI Listing
May 2019

Human Lactobacillus Strains from the Intestine can Suppress IgE-Mediated Degranulation of Rat Basophilic Leukaemia (RBL-2H3) Cells.

Microorganisms 2016 Oct 27;4(4). Epub 2016 Oct 27.

Department of Food Bioscience and Biotechnology, College of Bioresource Sciences, Nihon University, Fujisawa 252-8510, Japan.

Mast cells play a critical role in immunoglobulin E (IgE)-mediated allergic diseases, and the degranulation of mast cells is important in the pathogenesis of these diseases. A disturbance of the intestinal microflora, especially of endogenous lactic acid bacteria, might be a contributing factor for IgE-mediated allergic diseases. Additional knowledge regarding the interaction of human intestinal with mast cells is still necessary. Twenty-three strains of , including commercial and reference strains and strains from the human intestine, were tested for their ability to regulate degranulation of cells from rat basophilic leukemia RBL-2H3 cells (RBL-2H3) in vitro based on a β-hexosaminidase release assay. Each of the tested characteristically suppressed IgE-mediated degranulation of RBL-2H3 cells, and GG showed the strongest inhibitory effect on the cells. Furthermore, the bacteria isolated from the human intestine significantly suppressed degranulation of RBL-2H3 cellsin comparison with the reference strains. These results suggest that , particularly those from the human intestine, can affect the activation of mast cells in a strain-dependent manner. Further study should be conducted to analyse the understanding mechanism.
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http://dx.doi.org/10.3390/microorganisms4040040DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5192523PMC
October 2016

Probiotics modulate gut microbiota and health status in Japanese cedar pollinosis patients during the pollen season.

Eur J Nutr 2017 Oct 13;56(7):2245-2253. Epub 2016 Jul 13.

Functional Foods Forum, University of Turku, 20014, Turku, Finland.

Background: Japanese cedar pollinosis (JCP) is a challenging public health problem in Japan. Altered gut microbiota is associated with several diseases, including allergic diseases. However, only a few studies have focused on JCP and the underlying mechanisms for probiotic effects remain unclear. In addition, this study is the first observation of the correlation between the gut microbiota and blood lipid in JCP.

Methods: Faecal samples from JCP subjects were collected before and after treatment with (n = 14) and without (n = 11) LGG-TMC0356-fermented milk for 10 weeks. Gut microbiota composition was characterized from faecal DNA using sequencing of 16S rRNA genes.

Results: 16S rRNA-based operational taxonomic unit clustering of the microbiota revealed that LGG-TMC0356-fermented milk significantly altered gut microbiota after 10 weeks of milk consumption, and eight dominant genera of microbes were detected. During the JCP season, the Bacteroidetes/Firmicutes ratio, when compared to baseline, was significantly decreased in subjects at end of the study. Bacteroidetes showed positive correlation with LDL- and HDL-cholesterol levels, whereas Firmicutes showed negative correlation with total cholesterol, LDL- and HDL- cholesterol.

Conclusions: The altered gut microbiota through supplementation of fermented milk containing the study probiotics may be a prospective target for protection against JCP, with beneficial effects on blood lipid levels.
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http://dx.doi.org/10.1007/s00394-016-1264-3DOI Listing
October 2017

A combination of probiotics and whey proteins enhances anti-obesity effects of calcium and dairy products during nutritional energy restriction in aP2-agouti transgenic mice.

Br J Nutr 2015 Jun 14;113(11):1689-96. Epub 2015 Apr 14.

Nutrition Institute, The University of Tennessee,Knoxville,TN37996-1920,USA.

Lactobacillus rhamnosus GG, Lactobacillus paracasei TMC0409, Streptococcus thermophilus TMC1543 and whey proteins were used to prepare fermented milk. For the experiment aP2- agouti transgenic mice were pre-treated with a high-sucrose/high-fat diet for 6 weeks to induce obesity. The obese mice were fed a diet containing 1·2% Ca and either non-fat dried milk (NFDM) or probiotic-fermented milk (PFM) with nutritional energy restriction for 6 weeks. The animals were examined after the treatment for changes in body weight, fat pad weight, fatty acid synthase (FAS) activity, lypolysis, the expression levels of genes related to lipid metabolism, insulin sensitivity in adipocytes and skeletal muscle and the presence of biomarkers for oxidative and inflammatory stress in plasma. It was found that the PFM diet significantly reduced body weight, fat accumulation, and adipocyte FAS activity, and increased adipocyte lipolysis as compared with the effects of the NFDM diet (P<0·05). The adipose tissue gene expression of 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1) was significantly suppressed in mice that were fed PFM as compared with those that were fed NFDM (P<0·05). PFM caused a greater up-regulation of skeletal muscle PPARα, PPARδ, uncoupling protein 3 (UCP3) and GLUT4 expression and a significant decrease in the plasma concentration of insulin, malondialdehyde, TNF-α, monocyte chemotactic protein-1 and C-reactive protein as compared with the effects of NFDM (P<0·05). Fermentation of milk with selected probiotics and supplementation of milk with whey proteins may thus enhance anti-obesity effects of Ca and dairy products by the suppression of adipose tissue lipogenesis, activation of fat oxidation in skeletal muscle and reduction of oxidative and inflammatory stress.
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http://dx.doi.org/10.1017/S0007114515000914DOI Listing
June 2015

Influence of orally administered Lactobacillus GG on respiratory immune response in a murine model of diet-induced obesity.

Microbiol Immunol 2015 Feb;59(2):99-103

Technical Research Laboratory, Takanashi Milk Products, Yokohama, Kanagawa, 241-0023, Japan.

Mice with diet-induced obesity were fed with Lactobacillus rhamnosus GG (LGG) suspended in saline or saline alone (control mice). Pulmonary mRNA expression of IFN-γ; IFN-α receptor 1; CD247 antigen; killer cell lectin-like receptor subfamily K, member 1; TNF-α; IL-12 receptor β1 and IL-2 receptor β, and the proportion of Lactobacillales in feces were significantly greater in the LGG group than in the control mice (P < 0.05 and P < 0.01, respectively). These results suggest that LGG alters the respiratory immunity of obese subjects through having a potent impact on intestinal immunity.
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http://dx.doi.org/10.1111/1348-0421.12226DOI Listing
February 2015

Diversity of intestinal bifidobacteria in patients with Japanese cedar pollinosis and possible influence of probiotic intervention.

Curr Microbiol 2011 Jan 29;62(1):71-7. Epub 2010 May 29.

Technical Research Laboratory, Takanashi Milk Products Co., Ltd, Yokohama, Kanagawa, 241-0023, Japan.

This study was conducted to evaluate the potential association between intestinal bifidobacteria and Japanese cedar pollinosis (JCPsis) and possible influences of probiotic intervention. In this study, fecal samples were the collected from 29 JCPsis patients. The qualitative and quantitative analyses of fecal bifidobacteria were conducted by quantitative real-time PCR with 16S rRNA-gene-targeted species-specific primers before cedar pollen spread and after a 10-week intervention with fermented milk prepared with Lactobacillus GG and L. gasseri TMC0356 during pollen spread. Each JCPsis patient had a unique diversity of bifidobacteria, which varied qualitatively and quantitatively in an individual-dependent manner during pollen spread. The serum IgE concentration of JCPsis patients with more than 3 detectable Bifidobacterium species was significantly lower than that of patients with less than 2 detected species. The prevalence of B. adolescentis, B. longum, and B. catenulatum increased after probiotic intervention, although the changes were not statistically significant. These results suggest that lower diversity of intestinal Bifidobacterium species might be a pathological aspect of JCPsis. The diversity of intestinal bifidobacteria could be a prospective target for using probiotics in the management of IgE-mediated allergic disorders including JCPsis.
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http://dx.doi.org/10.1007/s00284-010-9667-5DOI Listing
January 2011

Bifidobacterium suppresses IgE-mediated degranulation of rat basophilic leukemia (RBL-2H3) cells.

Microbiol Immunol 2010 ;54(1):54-7

Technical Research Laboratory, Takanashi Milk Products Co. Ltd., 5 Honjyuku-cho, Asahi-ku, Yokohama, Kanagawa 241-0023, Japan.

Sixteen heat-killed bifidobacteria isolated from human intestine and a probiotic strain Lactobacillus GG were tested for their ability to influence IgE-mediated degranulation of rat basophilic leukemia (RBL-2H3) cells in vitro. The bifidobacteria suppressed IgE-mediated degranulation of RBL-2H3 cells by 1.6-56.4% in a strain-dependent manner. Bifidobacteria from healthy infants expressed high inhibitory effects on IgE-mediated degranulation (41-55%), while those from allergic infants varied greatly in their effects against degranulation. Bifidobacteria taxonomically identified as Bifidobacterium bifidum exhibited much stronger inhibitory effects against IgE-mediated degranulation than those taxonomically identified as B. adolescentis (P < 0.05).These results indicate that the intestinal bifidobacteria might be one of factors influencing IgE-mediated allergic responses.
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http://dx.doi.org/10.1111/j.1348-0421.2009.00185.xDOI Listing
April 2010

Lactobacillus strains stabilize intestinal microbiota in Japanese cedar pollinosis patients.

Microbiol Immunol 2009 Apr;53(4):198-205

Technical Research Laboratory, Takanashi Milk Products Co., Ltd, Yokohama, Kanagawa 241-0023, Japan.

A randomized double-blind, placebo-controlled trial was conducted to ascertain the intestinal microbiota-altering properties of LGG and L. gasseri TMC0356 (TMC0356) in Japanese cedar Cryptomeria japonica pollinosis patients. Fecal bacteria communities were examined before and after fermented milk administration using culture, FISH and T-RFLPmethods. Test group subjects showed the presence of LGG and TMC0356 along with a significant increase in fecal lactobacilli (P < 0.001) after giving LGG and TMC0356 fermented milk. Culture and FISH analysis revealed no significant changes in other intestinal bacterial groups. Each subject exhibited a characteristic T-RFLP profile pattern that varied quantitatively and qualitatively with JCP shedding. Profile changes were observed in 53% of placebo group subjects and in 21% of test group subject's post-administration, indicating that LGG and TMC0356 suppressed intestinal microbiota changes in JCPsis patients. The results suggest that intestinal microbiota might be more sensitive to exposure to environmental allergens than expected from the results of general culture method studies. Stabilization of intestinal microbiota by selected probiotic strains such as LGG and TMC0356 could be beneficial to homeostasis of the intestinal microbiota and useful in the management of JCPsis.
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http://dx.doi.org/10.1111/j.1348-0421.2009.00113.xDOI Listing
April 2009

Differentiated implication of Lactobacillus GG and L. gasseri TMC0356 to immune responses of murine Peyer's patch.

Microbiol Immunol 2009 Aug;53(8):475-80

Technical Research Laboratory, Takanashi Milk Products Co. Ltd, 5 Honjyuku-cho, Asahi-ku, Yokohama, Kanagawa 241-0023, Japan.

Lactobacillus GG and L. gasseri TMC0356 were examined for their potential to alter the immune responses of murine PP cells in vitro and in vivo. Lactobacillus GG and L. gasseri TMC0356 characteristically stimulated the production of IL-12, IL-6, IFN-gamma and IgA from isolated PP cells in vitro. Anatomical analysis indicated uptake of these bacteria by the PP tissue after giving orally in mice. Isolated PP cells exposed to Lactobacillus GG in vivo secreted more IFN-gamma, IL-6 and total IgA, whereas those exposed to L. gasseri TMC0356 in vivo did not exhibit altered immune responses in terms of cytokine and IgA production. Therefore, these two bacteria might exhibit different immunodulatory effects in host animals by strain-dependent interaction with gut-associated lymphoid tissues in vivo.
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http://dx.doi.org/10.1111/j.1348-0421.2009.00146.xDOI Listing
August 2009

Orally administrated Lactobacillus gasseri TMC0356 and Lactobacillus GG alleviated nasal blockage of guinea pig with allergic rhinitis.

Microbiol Immunol 2007 ;51(11):1109-14

Technical Research Laboratory, Takanashi Milk Products Co., Ltd., Yokohama, Japan.

Lactobacillus GG (LGG) and L. gasseri TMC0356 (TMC0356) were investigated for their ability to alleviate nasal blockage associated with allergic rhinitis using a guinea pig model. The increases in sRaw at 10 min and 5 hr after the exposure of the nasal mucosa to OVA were significantly alleviated in the guinea pigs orally administrated with LGG and TMC0356 compared with those of the control (P<0.05 and P<0.01). The total numbers of leukocytes, particularly eosinophils and neutrophils from the nasal cavity lavage fluid, and the OVA-specific IgE concentration in the serum were also decreased in the guinea pigs orally administrated with LGG and TMC0356, although the decreases were not statistically significant. These results suggest that LGG and TMC0356 can alleviate antigen-induced nasal blockage in earlyphase and late-phase inflammatory responses associated with allergic rhinitis.
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http://dx.doi.org/10.1111/j.1348-0421.2007.tb04006.xDOI Listing
May 2008
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