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2. Activation of hilA expression at low pH requires the signal sensor CpxA, but not the cognate response regulator CpxR, in Salmonella enterica serovar Typhimurium

Author

Nakayama, Shu-ichi, Kushiro, Akira, Asahara, Takashi, Tanaka, Ryu-ichiro, Hu, Lan, Kopecko, Dennis J., and Watanabe, Haruo

Subjects
Biosynthesis -- Analysis, Gene mutations -- Physiological aspects, Cells -- Genetic aspects, Genetic regulation -- Physiological aspects, Salmonella typhimurium -- Genetic aspects, Salmonella typhimurium -- Physiological aspects, Gene expression -- Physiological aspects, Shigella sonnei -- Genetic aspects, Shigella sonnei -- Physiological aspects, Virulence (Microbiology) -- Genetic aspects, Hydrogen-ion concentration -- Physiological aspects, Microbiology -- Research, Biological sciences
Abstract

A two-component regulatory system, cpxR-cpxA, plays an important role in the pH-dependent regulation of virF, a global activator for virulence determinants including invasion genes, in Shigella sonnei. The authors examined whether the cpxR-cpxA homologues have some function in the expression of Salmonella enterica serovar Typhimurium invasion genes via the regulation of hilA, an activator for these genes. In a Salmonella cpxA mutant, the hilA expression level was reduced to less than 10% of that in the parent strain at pH 6.0. This mutant strain also showed undetectable synthesis of an invasion gene product, SipC, at pH 6.0 and reduced cell invasion capacity--as low as 20% of that of the parent. In this mutant, the reduction in hilA expression was much less marked at pH 8.0 than at pH 6.0--no less than 50% of that in the parent, and no significant reduction was observed in either SipC synthesis or cell invasion rate, compared to the parent. Unexpectedly, a Salmonella cpxR mutant strain and the parent showed no apparent difference in all three characteristics described above at either pH. These results indicate that in Salmonella, the sensor kinase CpxA activates hilA, and consequently, invasion genes and cell invasion capacity at pH 6.0. At pH 8.0, however, CpxA does not seem to have a large role in activation of these factors. Further, the results show that this CpxA-mediated activation does not require its putative cognate response regulator, CpxR. This suggests that CpxA may interact with regulator(s) other than CpxR to achieve activation at low pH.

Published
2003

3. Dynamic analysis of human small intestinal microbiota after an ingestion of fermented milk by small-intestinal fluid perfusion using an endoscopic retrograde bowel insertion technique

Author

Takada, Toshihiko, primary, Chinda, Daisuke, additional, Mikami, Tatsuya, additional, Shimizu, Kensuke, additional, Oana, Kosuke, additional, Hayamizu, Shiro, additional, Miyazawa, Kuniaki, additional, Arai, Tetsu, additional, Katto, Miyuki, additional, Nagara, Yusuke, additional, Makino, Hiroshi, additional, Kushiro, Akira, additional, Oishi, Kenji, additional, and Fukuda, Shinsaku, additional

Published
2020
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4. Safety assessment of the candidate oral probiotic Lactobacillus crispatus YIT 12319: Analysis of antibiotic resistance and virulence-associated genes

Author

Terai, Tomohiko, primary, Kato, Kosuke, additional, Ishikawa, Eiji, additional, Nakao, Masumi, additional, Ito, Masahiko, additional, Miyazaki, Kouji, additional, Kushiro, Akira, additional, Imai, Susumu, additional, Nomura, Yoshiaki, additional, Hanada, Nobuhiro, additional, and Okumura, Takekazu, additional

Published
2020
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5. Colonization of the stratified squamous epithelium of the nonsecreting area of horse stomach by lactobacilli

Author

Ykui, Norikatsu, Shimazaki, Tomoko, Kushiro, Akira, Watanabe, Koichi, Uchida, Kazumi, Yuyama, Teruhiko, and Morotomi, Masami

Subjects
Digestive organs -- Microbiology, Horses -- Physiological aspects, Biological sciences
Abstract

Researchers have discovered several species of Lactobacillus that appear to exist symbiotically in the stomach of horses. Although they created a dense layer on the epithelium as is commonly seen in mice, rats, and pigs, many of the species adhered to horse epithelial cells but not to rat epithelial cells.

Published
2000

6. Safety assessment of the candidate oral probiotic Lactobacillus crispatus YIT 12319: Analysis of antibiotic resistance and virulence-associated genes

Author

Kato Kosuke, Susumu Imai, Nobuhiro Hanada, Yoshiaki Nomura, Masumi Nakao, Masahiko Ito, Tomohiko Terai, Takekazu Okumura, Kouji Miyazaki, Kushiro Akira, and Eiji Ishikawa

Subjects
medicine.drug_class, Antibiotics, Administration, Oral, Microbial Sensitivity Tests, Biology, Toxicology, Genome, Microbiology, law.invention, 03 medical and health sciences, Probiotic, 0404 agricultural biotechnology, Antibiotic resistance, law, medicine, Humans, Lactobacillus crispatus, Gene, 030304 developmental biology, Whole genome sequencing, Mouth, 0303 health sciences, Virulence, business.industry, Probiotics, Drug Resistance, Microbial, 04 agricultural and veterinary sciences, General Medicine, Food safety, biology.organism_classification, 040401 food science, Anti-Bacterial Agents, Genes, Bacterial, business, Food Science
Abstract

Lactobacillus crispatus YIT 12319 (LcY) was isolated from the oral cavity of a healthy subject as a new candidate probiotic with potential benefits for oral health. As a safety assessment of LcY, we performed an antibiotic susceptibility test and virulence-associated gene analysis using a draft genome sequence. Susceptibility to 15 antibiotics was analyzed according to the standard method of the International Dairy Federation/International Organization for Standardization, as recommended by the European Food Safety Authority. The results showed that the minimum inhibitory concentrations of LcY were not higher than those of other L. crispatus strains, which have not acquired resistance to any antibiotics, suggesting that LcY had no externally acquired transmissible antibiotic resistance genes. Analysis of virulence-associated genes using the draft genome of LcY found that there were fewer potential virulence-associated genes in LcY than in other probiotics. These findings suggest that LcY could be a candidate probiotic based on its safety profile.

Published
2020

7. Independent and Interactive Effects of Habitually Ingesting Fermented Milk Products Containing Lactobacillus casei Strain Shirota and of Engaging in Moderate Habitual Daily Physical Activity on the Intestinal Health of Older People

Author

Aoyagi, Yukitoshi, primary, Amamoto, Ryuta, additional, Park, Sungjin, additional, Honda, Yusuke, additional, Shimamoto, Kazuhito, additional, Kushiro, Akira, additional, Tsuji, Hirokazu, additional, Matsumoto, Hoshitaka, additional, Shimizu, Kensuke, additional, Miyazaki, Kouji, additional, Matsubara, Satoshi, additional, and Shephard, Roy J., additional

Published
2019
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12. A Single Species of Clostridium Subcluster XIVa Decreased in Ulcerative Colitis Patients

Author

Takeshita, Kozue, primary, Mizuno, Shinta, additional, Mikami, Yohei, additional, Sujino, Tomohisa, additional, Saigusa, Keiichiro, additional, Matsuoka, Katsuyoshi, additional, Naganuma, Makoto, additional, Sato, Tadashi, additional, Takada, Toshihiko, additional, Tsuji, Hirokazu, additional, Kushiro, Akira, additional, Nomoto, Koji, additional, and Kanai, Takanori, additional

Published
2016
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15. Early-Life Events, Including Mode of Delivery and Type of Feeding, Siblings and Gender, Shape the Developing Gut Microbiota

Author

Martin, Rocio, primary, Makino, Hiroshi, additional, Cetinyurek Yavuz, Aysun, additional, Ben-Amor, Kaouther, additional, Roelofs, Mieke, additional, Ishikawa, Eiji, additional, Kubota, Hiroyuki, additional, Swinkels, Sophie, additional, Sakai, Takafumi, additional, Oishi, Kenji, additional, Kushiro, Akira, additional, and Knol, Jan, additional

Published
2016
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16. Fermented Milk Containing Lactobacillus casei Strain Shirota Preserves the Diversity of the Gut Microbiota and Relieves Abdominal Dysfunction in Healthy Medical Students Exposed to Academic Stress

Author

Kato-Kataoka, Akito, primary, Nishida, Kensei, additional, Takada, Mai, additional, Kawai, Mitsuhisa, additional, Kikuchi-Hayakawa, Hiroko, additional, Suda, Kazunori, additional, Ishikawa, Hiroshi, additional, Gondo, Yusuke, additional, Shimizu, Kensuke, additional, Matsuki, Takahiro, additional, Kushiro, Akira, additional, Hoshi, Ryoutaro, additional, Watanabe, Osamu, additional, Igarashi, Tomoki, additional, Miyazaki, Kouji, additional, Kuwano, Yuki, additional, and Rokutan, Kazuhito, additional

Published
2016
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17. Mother-to-Infant Transmission of Intestinal Bifidobacterial Strains Has an Impact on the Early Development of Vaginally Delivered Infant's Microbiota

Author

Makino, Hiroshi, primary, Kushiro, Akira, additional, Ishikawa, Eiji, additional, Kubota, Hiroyuki, additional, Gawad, Agata, additional, Sakai, Takafumi, additional, Oishi, Kenji, additional, Martin, Rocio, additional, Ben-Amor, Kaouther, additional, Knol, Jan, additional, and Tanaka, Ryuichiro, additional

Published
2013
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18. Ethnic diversity of gut microbiota: Species characterization of Bacteroides fragilis group and genus Bifidobacterium in healthy Belgian adults, and comparison with data from Japanese subjects

Author

Ishikawa, Eiji, primary, Matsuki, Takahiro, additional, Kubota, Hiroyuki, additional, Makino, Hiroshi, additional, Sakai, Takafumi, additional, Oishi, Kenji, additional, Kushiro, Akira, additional, Fujimoto, Junji, additional, Watanabe, Koichi, additional, Watanuki, Masaaki, additional, and Tanaka, Ryuichiro, additional

Published
2013
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20. Transmission of Intestinal Bifidobacterium longum subsp. longum Strains from Mother to Infant, Determined by Multilocus Sequencing Typing and Amplified Fragment Length Polymorphism

Author

Makino, Hiroshi, primary, Kushiro, Akira, additional, Ishikawa, Eiji, additional, Muylaert, Delphine, additional, Kubota, Hiroyuki, additional, Sakai, Takafumi, additional, Oishi, Kenji, additional, Martin, Rocio, additional, Ben Amor, Kaouther, additional, Oozeer, Raish, additional, Knol, Jan, additional, and Tanaka, Ryuichiro, additional

Published
2011
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26. Immunogenicity and protective efficacy of orally administered recombinant Lactococcus lactis expressing surface-bound HIV Env

Author

Xin, Ke-Qin, primary, Hoshino, Yuka, additional, Toda, Yoshihiko, additional, Igimi, Shizunobu, additional, Kojima, Yoshitsugu, additional, Jounai, Nao, additional, Ohba, Kenji, additional, Kushiro, Akira, additional, Kiwaki, Mayumi, additional, Hamajima, Kenji, additional, Klinman, Dennis, additional, and Okuda, Kenji, additional

Published
2003
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32. Fermented milk containing Lactobacillus casei strain Shirota reduces incidence of hard or lumpy stools in healthy population.

Author

Sakai, Takafumi, Makino, Hiroshi, Ishikawa, Eiji, Oishi, Kenji, and Kushiro, Akira

Subjects
LACTOBACILLUS casei, FERMENTED milk, PROBIOTICS, CONSTIPATION, RANDOMIZED controlled trials, DEFECATION disorders, PROBABILITY theory
Abstract

The objective of the present study was to investigate the efficacy of fermented milk containing Lactobacillus casei strain Shirota (LcS) in a healthy population. Healthy subjects with Bristol Stool Form Scale (BS) score <  3.0 were randomized to fermented milk treatment for 3 weeks or non-intervention control. The primary endpoint was the proportion of subjects that produced hard or lumpy stools (HLS) ≥≥  25% of bowel movements (H-HLS). Secondary endpoints included changes in BS score, constipation-related symptom scores and stool parameters. Efficacy was analyzed in 39 subjects. After 3 weeks of treatment the proportion of H-HLS subjects had significantly decreased from 73.7% to 36.8%, whereas in the control group the proportion had increased from 75.0% to 85.0% during the same period ( P  ==  0.002). The BS score was significantly improved after the treatment compared with the control ( P  <  0.001). In conclusion, daily consumption of fermented milk containing LcS reduced the incidence of HLS. [ABSTRACT FROM AUTHOR]

Published
2011
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34. Molecular cloning and sequence determination of the tuf gene coding for the elongation factor Tu of Thermus thermophilus HB8.

Author

Kushiro, Akira, Shimizu, Masato, and Tomita, Ken-Ichi

Subjects
*ORGANIC acids, *ESCHERICHIA coli, *AMINO acid sequence, *ENTEROBACTERIACEAE, *SACCHAROMYCES cerevisiae, *HOMOLOGY (Biology)
Abstract

The tuf gene, which encodes the elongation factor Tu (EF-Tu) of Thermus thermophilus HB8, and its flanking regions were cloned and sequenced. The gene encoding EF-G was found upstream of the 5' end of the tuf gene. The tuf gene of T. thermophilus HB8 had a very high G-C content and 84.5% of the third base in codon usage was either G or C. The deduced primary structure of the EF-Tu was composed of 405 amino acid residues with a Mr = 44658. A comparison of the amino acid sequence of EF-Tu from T. thermophilus HB8 with those of Escherichia coli and Saccharomyces cerevisiae mutochondria showed a very high sequence homology (65 - 70%). Two Cys residues out of the three found in E. coli EF-Tu had been replaced with Val in T. thermophilus HB8 EF-Tu. An extra amino acid sequence of ten residues, consisting predominantly of basic amino acids (Met-182-Gly-191), which does not occur in EF-Tu of E. coli, was found in T. thermophilus HB8. [ABSTRACT FROM AUTHOR]

Published
1987
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35. Immunogenicity and protective efficacy of orally administered recombinant Lactococcus lactisexpressing surface-bound HIV Env

Author

Xin, Ke-Qin, Hoshino, Yuka, Toda, Yoshihiko, Igimi, Shizunobu, Kojima, Yoshitsugu, Jounai, Nao, Ohba, Kenji, Kushiro, Akira, Kiwaki, Mayumi, Hamajima, Kenji, Klinman, Dennis, and Okuda, Kenji

Abstract

This study investigates whether genetically modified orally administered Lactococcus lactis(L lactis) could be used as an HIV vaccine. L lactisis immunogenic and extremely safe when delivered orally. We created a recombinant L lactisvector expressing the envelope protein of HIV on its cell surface. Oral immunization with this vector induced high levels of HIV-specific serum IgG and fecal IgA antibodies. Cell-mediated immune responses also were generated in both the regional lymph nodes and the spleen. Dendritic cells are readily infected by L lactisand appear to play a potential role in mediating the development of these immune responses. The protective efficacy of this vaccine strategy was demonstrated by challenging mice intraperitoneally with an HIV Env–expressing vaccinia virus. Their viral loads were 350-fold lower than those of control mice. These findings support the further development of L lactis–based HIV vaccines. (Blood. 2003; 102:223-228)

Published
2003
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36. Molecular cloning, nucleotide sequence and expression of the tufBgene encoding elongation factor Tu from Thermus thermophilusHB8

Author

Satoh, Motohide, Tanaka, Toshikazu, Kushiro, Akira, Hakoshima, Toshio, and Tomita, Ken-ichi

Abstract

The tufBgene encoding elongation factor Tu (EF-Tu) of Thermus thermophilusHB8 was cloned and expressed. Compared with the known tufAgene of T. thermophilus, nucleotide differences were found at 10 positions out of 1221 nucleotides, and amino acid substitutions were found at 4 positions out of 406 amino acids. The tufBproduct was 70.9% homologous to the corresponding sequence of the tufBproduct of E. coli. The G+C content of the third base of the codon in the tufBgene was 84.8% and G was especially preferred in this position.

Published
1991
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37. A PROBIOTIC FERMENTED MILK DRINK CONTAINING LACTOBACILLUS CASEI STRAIN SHIROTA IMPROVES STOOL CONSISTENCY OF SUBJECTS WITH HARD STOOLS.

Author

Tilley, Linde, Keppens, Kadeen, Kushiro, Akira, Takada, Toshihiko, Sakai, Takafiimi, Vaneechoutte, Mario, and Degeest, Bart

Subjects
*PROBIOTICS, *LACTOBACILLUS casei, *LACTOBACILLUS, *FERMENTED milk, *MILK substitutes
Abstract

The aim of this study was to investigate the effect of a fermented milk drink containing Lactobacillus casei Shirota (LcS) on the bowel habit (with emphasis on stool consistency) o f subjects suffering from hard stools. Secondly, it was tested whether the probiotic strain (LcS), was able to survive throughout the gastrointestinal tract. A double-blind, placebo-controlled, randomized study was carried out over an eight-week period in subjects with symptoms o f constipation (n=120). To all subjects, 65 ml/day of a probiotic fermented milk drink containing LcS or a placebo was administered. Patients completed a questionnaire to assess the consistency o f their stools. In half of the study population, the survival of the probiotic strain (LcS) was tested using (i) culture and (ii) an LcS specific monoclonal antibody to identify the cultured colonies as LcS (ELISA). There was a significant decrease in stool hardening when consuming a fermented milk drink containing LcS. The observed clinical effect went hand in hand with the observed microbiological effect as the number of viable LcS bacteria in the faeces increased when consuming fermented milk drink containing LcS and decreased during wash-out. [ABSTRACT FROM AUTHOR]

Published
2014

38. Probiotic Lactobacillus casei activates innate immunity via NF-kappaB and p38 MAP kinase signaling pathways.

Author

Kim YG, Ohta T, Takahashi T, Kushiro A, Nomoto K, Yokokura T, Okada N, and Danbara H

Subjects
Animals, Cells, Cultured, Cytokines biosynthesis, Fibrosarcoma immunology, Hot Temperature, Intracellular Signaling Peptides and Proteins metabolism, Listeria monocytogenes, Listeriosis immunology, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Microbial Viability, NF-kappa B metabolism, Neoplasms, Experimental immunology, Nod2 Signaling Adaptor Protein, Severe Combined Immunodeficiency, Signal Transduction, Spleen cytology, Spleen immunology, Spleen metabolism, Toll-Like Receptor 2 metabolism, p38 Mitogen-Activated Protein Kinases metabolism, Immunity, Innate, Lacticaseibacillus casei physiology, NF-kappa B immunology, p38 Mitogen-Activated Protein Kinases immunology
Abstract

Probiotic bacteria are microorganisms that benefit the host through improvement of the balance of intestinal microflora and possibly by augmentation of host defense systems. We examined the mechanisms for the up-regulation of innate immune responses by a probiotic Lactobacillus casei ATCC27139, in vivo. Using mouse models of systemic Listeria monocytogenes infection and MethA fibrosarcoma tumorigenesis in combination with BALB/c and SCID mice, we found that parenteral administration of L. casei ATCC27139 confers a protective effect against L. monocytogenes infection and anti-tumor activity against MethA fibrosarcoma by activation of innate immunity, while L. casei ATCC27139-J1R strains, which are J1 phage-resistant strains that have been selected from MNNG-treated clones, lacked these activities. Substantial differences between ATCC27139 and ATCC27139-J1R strains were observed in the capacity to induce innate cytokines such as TNF-alpha, IL-12, IL-18, and IFN-gamma, and pathogen-associated molecular pattern receptors, TLR2 and Nod2, by spleen cells. In addition, although phosphorylation of NF-kappaB p65 in spleen was equally enhanced in the ATCC27139- and the ATCC27139-J1R-treated groups, phosphorylation of both p38 MAPK and MAPKAPK-2 was significantly induced only by ATCC27139. Furthermore, inhibitors of NF-kappaB (sulfasalazine) and p38 MAPK (SB203580) significantly reduced cytokine production by the spleen cells of the mice treated with L. casei ATCC27139, suggesting that both NF-kappaB and p38 MAPK signaling pathways play important roles in the augmentation of innate immunity by the probiotic L. casei.

Published
2006
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