Gifu

Gifu-shi, Japan
Gifu-shi, Japan
SEARCH FILTERS
Time filter
Source Type

TOKYO, May 23, 2017 /PRNewswire/ -- VT Holdings (TOKYO: 7593) is pleased to announce its results for the fiscal year ended March 31, 2017. During the year ended March 31, 2017, unit sales of new cars were up 2.8% year on year. The Group was affected by Nissan Motor Company's suspension of sales of two light vehicle models between April and June 2016, but higher sales by the Group's 12 subsidiaries prompted increased sales by the Group. These subsidiaries include Motoren Shizuoka (began sales as a BMW dealer in April 2016), Wessex Garages Holding Limited of the United Kingdom (became a subsidiary in May) and Master Automocion, S.L. of Spain (became a subsidiary in October). Unit sales of new and used cars amounted to 82,916 vehicles, up 9,099 units, or 12.3%, year on year. During the year, VT Holdings recorded consolidated net sales of JPY169,560 million (up 15.8% YoY), operating income of JPY7,592 million (down 0.4% YoY), ordinary income of JPY7,937 million (up 4.4% YoY) and profit attributable to owners of parent of JPY4,421 million (up 8.1% YoY). In the automotive business, the Company's sales amounted to JPY162,687 million (up 16.2% YoY), with operating income of JPY7,529 million (down 2.4% YoY). Within the automotive business, in the new car segment sales in Japan struggled, but overseas and other Group companies exceeded the previous year's sales on a unit basis, leading to sales of 33,616 units (up 22.0% YoY), contributing to higher sales and income. In the automotive business's used car segment, overseas exports were lackluster, but sales by newly consolidated subsidiaries expanded. In this segment, unit sales for the Group accordingly amounted to 49,300 units (up 6.6% YoY). The higher unit sales lifted segment sales, but income declined. In the automotive business's service segment, existing and newly consolidated subsidiaries experienced increases in orders for inspections and repairs, as well as commission revenue, leading to higher sales and income. Sales and income also rose in the rent-a-car segment. In the housing-related business, sales came to JPY6,731 million (up 7.4% YoY), and operating income was JPY541 million (up 86.2% YoY). In the housing-related business, the Company is developing the condominium business in Aichi and Gifu prefectures, where performance was extremely strong. Business was also robust overall in the detached homes segment, which the Company is expanding at its locations in Tokyo, Osaka and Nagoya, and the company worked to increase orders from commercial facilities outside the Group. For the fiscal year ending March 31, 2018, VT Holdings forecasts net sales of JPY196.0 billion (up 15.6% YoY), operating income of JPY8.5 billion (up 12.0% YoY), ordinary income of JPY8.5 billion (up 7.1% YoY) and profit attributable to owners of parent of JPY4.8 billion (up 8.5% YoY). The Company forecasts ordinary dividends for the year of JPY18 per share, comprising interim and year-end dividends of JPY9 each. VT Holdings Co., Ltd. (7593, First Section, TSE) "Summary of Consolidated Financial Results for the Year Ended March 31, 2017" is available here: http://www.vt-holdings.co.jp/eng/ir/library/pdf/20170523_FS.pdf This release is for the purpose of providing information to serve as a reference for investment decisions and not for the purpose of soliciting investment. Please make your own judgment on final decisions such as investment policy, timing and selection. Please be advised that we do not assume any responsibility for damages caused by this service. Borderless IR specializes in the overseas distribution of IR content, including the dissemination of newsletters and annual reports providing the latest information and main strengths of Japanese companies directly to overseas investors through leading global media, corporate information database services and mailing lists. Borderless is also engaged in supporting other global IR efforts. To view the original version on PR Newswire, visit:http://www.prnewswire.com/news-releases/vt-holdings-co-ltd-7593-first-section-tokyo-stock-exchange-issues-operating-performance-for-the-fiscal-year-ended-march-31-2017-300461968.html


During the year ended March 31, 2017, unit sales of new cars were up 2.8% year on year. The Group was affected by Nissan Motor Company's suspension of sales of two light vehicle models between April and June 2016, but higher sales by the Group's 12 subsidiaries prompted increased sales by the Group. These subsidiaries include Motoren Shizuoka (began sales as a BMW dealer in April 2016), Wessex Garages Holding Limited of the United Kingdom (became a subsidiary in May) and Master Automocion, S.L. of Spain (became a subsidiary in October). Unit sales of new and used cars amounted to 82,916 vehicles, up 9,099 units, or 12.3%, year on year. During the year, VT Holdings recorded consolidated net sales of JPY169,560 million (up 15.8% YoY), operating income of JPY7,592 million (down 0.4% YoY), ordinary income of JPY7,937 million (up 4.4% YoY) and profit attributable to owners of parent of JPY4,421 million (up 8.1% YoY). In the automotive business, the Company's sales amounted to JPY162,687 million (up 16.2% YoY), with operating income of JPY7,529 million (down 2.4% YoY). Within the automotive business, in the new car segment sales in Japan struggled, but overseas and other Group companies exceeded the previous year's sales on a unit basis, leading to sales of 33,616 units (up 22.0% YoY), contributing to higher sales and income. In the automotive business's used car segment, overseas exports were lackluster, but sales by newly consolidated subsidiaries expanded. In this segment, unit sales for the Group accordingly amounted to 49,300 units (up 6.6% YoY). The higher unit sales lifted segment sales, but income declined. In the automotive business's service segment, existing and newly consolidated subsidiaries experienced increases in orders for inspections and repairs, as well as commission revenue, leading to higher sales and income. Sales and income also rose in the rent-a-car segment. In the housing-related business, sales came to JPY6,731 million (up 7.4% YoY), and operating income was JPY541 million (up 86.2% YoY). In the housing-related business, the Company is developing the condominium business in Aichi and Gifu prefectures, where performance was extremely strong. Business was also robust overall in the detached homes segment, which the Company is expanding at its locations in Tokyo, Osaka and Nagoya, and the company worked to increase orders from commercial facilities outside the Group. For the fiscal year ending March 31, 2018, VT Holdings forecasts net sales of JPY196.0 billion (up 15.6% YoY), operating income of JPY8.5 billion (up 12.0% YoY), ordinary income of JPY8.5 billion (up 7.1% YoY) and profit attributable to owners of parent of JPY4.8 billion (up 8.5% YoY). The Company forecasts ordinary dividends for the year of JPY18 per share, comprising interim and year-end dividends of JPY9 each. VT Holdings Co., Ltd. (7593, First Section, TSE) "Summary of Consolidated Financial Results for the Year Ended March 31, 2017" is available here: http://www.vt-holdings.co.jp/eng/ir/library/pdf/20170523_FS.pdf This release is for the purpose of providing information to serve as a reference for investment decisions and not for the purpose of soliciting investment. Please make your own judgment on final decisions such as investment policy, timing and selection. Please be advised that we do not assume any responsibility for damages caused by this service. Borderless IR specializes in the overseas distribution of IR content, including the dissemination of newsletters and annual reports providing the latest information and main strengths of Japanese companies directly to overseas investors through leading global media, corporate information database services and mailing lists. Borderless is also engaged in supporting other global IR efforts. To view the original version on PR Newswire, visit:http://www.prnewswire.com/news-releases/vt-holdings-co-ltd-7593-first-section-tokyo-stock-exchange-issues-operating-performance-for-the-fiscal-year-ended-march-31-2017-300461968.html


News Article | November 10, 2016
Site: www.acnnewswire.com

The ninth HKTDC Hong Kong International Wine & Spirits Fair opened today and continues through 12 November at the Hong Kong Convention and Exhibition Centre (HKCEC). This morning's opening ceremony was officiated by Gregory So, Secretary for Commerce and Economic Development of the Hong Kong Special Administrative Region (HKSAR) Government and Philip Yung, Permanent Secretary for Commerce and Economic Development (Commerce, Industry and Tourism) of the HKSAR Government. Speaking at the opening ceremony, Benjamin Chau, Acting Executive Director, Hong Kong Trade Development Council (HKTDC), highlighted the diverse characteristics of the International Wine & Spirits Fair. "Featuring more than 1,060 exhibitors from 37 countries and regions, the Wine & Spirits Fair is an effective international promotion platform. The success of the fair is due to a variety of factors: Zero duties on Hong Kong wine imports since 2008, a large international exhibitor presence, international buyers especially wine importers from Asia, high value-added business opportunities and networking activities including grand tasting sessions, master classes, wine tastings, cocktail demonstrations as well as seminars," Mr Chau said. Strong international flavour at the fair Since the HKSAR Government scrapped import duties on wine in 2008, the wine industry has recorded tremendous growth, attracting industry players to start or expand their business in Hong Kong. The value of Hong Kong's wine imports rose from HK$1.6 billion in 2007 to HK$10.8 billion in 2015, a more than six-fold increase. Being a well-known wine trading and distributing hub, wine exporting countries are seeking to tap into the Asian market through Hong Kong. Besides Croatia, Finland and the Philippines exhibiting at the fair for the first time, wine producing regions, wine associations and trade commissions from around the world have formed 30 pavilions to promote their products. Among them, first-time group pavilion organisers include the Azerbaijan Export and Investment Promotion Foundation, Bulgarian Wine Export Association, Economic and Information Technology Commission of Guizhou Province from the Chinese mainland, Fukushima Prefectural Government and Kyushu Shochu Culture & Tourism from Japan, FENADEGAS from Portugal, the Distilled Spirits Council of the United States, and the Ministry of Agriculture, Forestry and Food of the Republic of Slovenia. Located in South Central Europe, Slovenia is a wine producing country less familiar to consumers in Hong Kong and Asia. Slovenia's viniculture is characterised by the country's diverse geography and microclimates; its latitude aligns with many renowned and prolific wine-producing regions like Bordeaux, Burgundy and Northern Rhone. Around 70 per cent of Slovenian wines qualify as premium wine. Aiming to capture the attention of Asia's developing markets through Hong Kong, Dejan Zidan, Deputy Prime Minister and Minister of Agriculture, Forestry and Food of the Republic of Slovenia, hosted today's presentation under the theme of "Discover Excellent Wines From Slovenia - Taste the Slovenian Identity". While Slovenia has a particularly high profile at the fair this year, a world of exquisite wines from around the world are also on show including: - Wine from Israel, a country with 5,000 years of wine-making history. Produced by Hevron Heights, Armagedon (Booth no.: 3E-D11) is brewed using traditional methods. Using grapes grown on the Judean Mountains at high altitude (950m) and aged for 24 months in French oak barrels, Armagedon is regarded as kosher wine, produced in accordance with Judaism's religious laws. - Bulgarian orange wine from Wine Cellar Villa Melnik Ltd (Booth no.: 3D-B26). Orange wine, in spite of its name, is not made from oranges. Its darker colour results from extended contact of white grape juice with grape skins over a longer period of time. Orange wine is intense with a dry, tannic taste and nuttiness derived from oxidation, and can be paired with a wide variety of dishes ranging from beef to fish. - Crown Royal's Northern Harvest Rye from Canada (Booth no.: 3CON-064). Crown Royal's Northern Harvest Rye stunned the whisky world as the first Canadian whisky to earn a title in the authoritative Jim Murray's Annual Whisky Bible with almost full marks and was named World Whisky of the Year 2016. In addition to zones such as Wine & Liquor Products, Whisky and Spirits and Friends of Wine, where the perfect food pairings are showcased, there are also dedicated zones promoting the industry's all-round developments, such as Wine Investment, Wine Education and Wine Storage & Logistics zones. Promoting industry interaction During the Wine & Spirits Fair, more than 70 special events are arranged to provide a comprehensive platform for trading and exchange. These include the Wine Industry Conference, gala dinner, tasting sessions, master classes and thematic seminars. Close to 50 wine tasting sessions are organised to spotlight wines from Austria, Australia, Bulgaria, France, Germany, Guizhou (Chinese mainland), Japan, Mexico, Portugal, Slovenia, Spain and the US. Today's Wine Industry Conference is titled "Uncover the Opportunities of the New Cool Climate Wine Trend". Meanwhile, the eighth edition of the Cathay Pacific Hong Kong International Wine & Spirit Competition Award Presentation Ceremony will be held tonight. Following the cocktail reception, the Gala Dinner titled "I FEEL SLOVENIA" will feature a menu prepared by Janez Bratovz, head chef of celebrated Slovenian restaurant JB RESTAVRACIJA. The seminar "How to Reach the Right Customers in China" and the buyer forum "Uncovering Business Opportunities in Booming Markets of Wine and Spirits" will be held tomorrow afternoon for the industry to discuss hot topics. Public Day On Saturday (12 November), the fair will be open to members of the public aged 18 or above, with tickets priced at HK$200*. Public visitors with full-priced tickets on that day will receive a Lucaris crystal wine glass valued at HK$110 on a first-come first-served basis while stocks last. Two master classes will be held on the public day including "Understanding Quality in Wines Currently Trending around the World with Jeannie Cho Lee MW" and "Sensory Experience of Wine by Debra Meiburg MW". The public are also welcome to join wine tasting sessions, cocktail, whisky and spirit demonstrations and seminars. These include "Gifu Sake and Pottery Appreciation", "Enjoy Shochu from Kyushu with Kumamon", "Choosing from a Wine List - Tips and Tricks" and "Hong Kong Inter-University Wine Challenge 2016". This year's fair once again headlines the Hong Kong Wine Journey citywide promotion, which encompasses a series of wine tastings, wine and food menu pairing, seminars, themed tours and Lan Kwai Fong carnival. More than 160 restaurants will feature promotions such as "Birthday Wine" and "Wine and Food Pairing Menu". For more details, please refer to the Hong Kong Wine Journey map or the website. Wine business keeps flowing through Hong Kong In the first nine months of 2016, Hong Kong's wine imports reached HK$9.1 billion, a 22 per cent year-on-year increase. As for the city's exports, they totalled HK$4.1 billion, up 25 per cent over the same period last year. *Tickets: Members of the public can purchase Public Day admission tickets on site priced at HK$200. Tickets for Public Day master classes are priced at HK$350 (including admission) and are available on a first-come first-served basis. Fair Website: www.hktdc.com/hkwinefair Social Media Hashtag: #HKWineandSpirits Photo Download: http://bit.ly/2fA4drM Media Registration: Media may register on-site with their business cards and/or media identification. To view press releases in Chinese, please visit http://mediaroom.hktdc.com/tc About HKTDC Established in 1966, the Hong Kong Trade Development Council (HKTDC) is a statutory body dedicated to creating opportunities for Hong Kong's businesses. With more than 40 offices globally, including 13 on the Chinese mainland, the HKTDC promotes Hong Kong as a platform for doing business with China, Asia and the world. With 50 years of experience, the HKTDC organises international exhibitions, conferences and business missions to provide companies, particularly SMEs, with business opportunities on the mainland and in international markets, while providing information via trade publications, research reports and digital channels including the media room. For more information, please visit: www.hktdc.com/aboutus. Follow us on Google+, Twitter @hktdc, LinkedIn. Google+: https://plus.google.com/+hktdc Twitter: http://www.twitter.com/hktdc LinkedIn: http://www.linkedin.com/company/hong-kong-trade-development-council Contact:


News Article | November 10, 2016
Site: www.newsmaker.com.au

HONG KONG - The ninth HKTDC Hong Kong International Wine & Spirits Fair opened today and continues through 12 November at the Hong Kong Convention and Exhibition Centre (HKCEC). This morning's opening ceremony was officiated by Gregory So, Secretary for Commerce and Economic Development of the Hong Kong Special Administrative Region (HKSAR) Government and Philip Yung, Permanent Secretary for Commerce and Economic Development (Commerce, Industry and Tourism) of the HKSAR Government. Speaking at the opening ceremony, Benjamin Chau, Acting Executive Director, Hong Kong Trade Development Council (HKTDC), highlighted the diverse characteristics of the International Wine & Spirits Fair. "Featuring more than 1,060 exhibitors from 37 countries and regions, the Wine & Spirits Fair is an effective international promotion platform. The success of the fair is due to a variety of factors: Zero duties on Hong Kong wine imports since 2008, a large international exhibitor presence, international buyers especially wine importers from Asia, high value-added business opportunities and networking activities including grand tasting sessions, master classes, wine tastings, cocktail demonstrations as well as seminars," Mr Chau said. Since the HKSAR Government scrapped import duties on wine in 2008, the wine industry has recorded tremendous growth, attracting industry players to start or expand their business in Hong Kong. The value of Hong Kong's wine imports rose from HK$1.6 billion in 2007 to HK$10.8 billion in 2015, a more than six-fold increase. Being a well-known wine trading and distributing hub, wine exporting countries are seeking to tap into the Asian market through Hong Kong. Besides Croatia, Finland and the Philippines exhibiting at the fair for the first time, wine producing regions, wine associations and trade commissions from around the world have formed 30 pavilions to promote their products. Among them, first-time group pavilion organisers include the Azerbaijan Export and Investment Promotion Foundation, Bulgarian Wine Export Association, Economic and Information Technology Commission of Guizhou Province from the Chinese mainland, Fukushima Prefectural Government and Kyushu Shochu Culture & Tourism from Japan, FENADEGAS from Portugal, the Distilled Spirits Council of the United States, and the Ministry of Agriculture, Forestry and Food of the Republic of Slovenia. Located in South Central Europe, Slovenia is a wine producing country less familiar to consumers in Hong Kong and Asia. Slovenia's viniculture is characterised by the country's diverse geography and microclimates; its latitude aligns with many renowned and prolific wine-producing regions like Bordeaux, Burgundy and Northern Rhone. Around 70 per cent of Slovenian wines qualify as premium wine. Aiming to capture the attention of Asia's developing markets through Hong Kong, Dejan Zidan, Deputy Prime Minister and Minister of Agriculture, Forestry and Food of the Republic of Slovenia, hosted today's presentation under the theme of "Discover Excellent Wines From Slovenia - Taste the Slovenian Identity". While Slovenia has a particularly high profile at the fair this year, a world of exquisite wines from around the world are also on show including: - Wine from Israel, a country with 5,000 years of wine-making history. Produced by Hevron Heights, Armagedon (Booth no.: 3E-D11) is brewed using traditional methods. Using grapes grown on the Judean Mountains at high altitude (950m) and aged for 24 months in French oak barrels, Armagedon is regarded as kosher wine, produced in accordance with Judaism's religious laws. - Bulgarian orange wine from Wine Cellar Villa Melnik Ltd (Booth no.: 3D-B26). Orange wine, in spite of its name, is not made from oranges. Its darker colour results from extended contact of white grape juice with grape skins over a longer period of time. Orange wine is intense with a dry, tannic taste and nuttiness derived from oxidation, and can be paired with a wide variety of dishes ranging from beef to fish. - Crown Royal's Northern Harvest Rye from Canada (Booth no.: 3CON-064). Crown Royal's Northern Harvest Rye stunned the whisky world as the first Canadian whisky to earn a title in the authoritative Jim Murray's Annual Whisky Bible with almost full marks and was named World Whisky of the Year 2016. In addition to zones such as Wine & Liquor Products, Whisky and Spirits and Friends of Wine, where the perfect food pairings are showcased, there are also dedicated zones promoting the industry's all-round developments, such as Wine Investment, Wine Education and Wine Storage & Logistics zones. During the Wine & Spirits Fair, more than 70 special events are arranged to provide a comprehensive platform for trading and exchange. These include the Wine Industry Conference, gala dinner, tasting sessions, master classes and thematic seminars. Close to 50 wine tasting sessions are organised to spotlight wines from Austria, Australia, Bulgaria, France, Germany, Guizhou (Chinese mainland), Japan, Mexico, Portugal, Slovenia, Spain and the US. Today's Wine Industry Conference is titled "Uncover the Opportunities of the New Cool Climate Wine Trend". Meanwhile, the eighth edition of the Cathay Pacific Hong Kong International Wine & Spirit Competition Award Presentation Ceremony will be held tonight. Following the cocktail reception, the Gala Dinner titled "I FEEL SLOVENIA" will feature a menu prepared by Janez Bratovz, head chef of celebrated Slovenian restaurant JB RESTAVRACIJA. The seminar "How to Reach the Right Customers in China" and the buyer forum "Uncovering Business Opportunities in Booming Markets of Wine and Spirits" will be held tomorrow afternoon for the industry to discuss hot topics. On Saturday (12 November), the fair will be open to members of the public aged 18 or above, with tickets priced at HK$200*. Public visitors with full-priced tickets on that day will receive a Lucaris crystal wine glass valued at HK$110 on a first-come first-served basis while stocks last. Two master classes will be held on the public day including "Understanding Quality in Wines Currently Trending around the World with Jeannie Cho Lee MW" and "Sensory Experience of Wine by Debra Meiburg MW". The public are also welcome to join wine tasting sessions, cocktail, whisky and spirit demonstrations and seminars. These include "Gifu Sake and Pottery Appreciation", "Enjoy Shochu from Kyushu with Kumamon", "Choosing from a Wine List - Tips and Tricks" and "Hong Kong Inter-University Wine Challenge 2016". This year's fair once again headlines the Hong Kong Wine Journey citywide promotion, which encompasses a series of wine tastings, wine and food menu pairing, seminars, themed tours and Lan Kwai Fong carnival. More than 160 restaurants will feature promotions such as "Birthday Wine" and "Wine and Food Pairing Menu". For more details, please refer to the Hong Kong Wine Journey map or the website. In the first nine months of 2016, Hong Kong's wine imports reached HK$9.1 billion, a 22 per cent year-on-year increase. As for the city's exports, they totalled HK$4.1 billion, up 25 per cent over the same period last year. Members of the public can purchase Public Day admission tickets on site priced at HK$200. Tickets for Public Day master classes are priced at HK$350 (including admission) and are available on a first-come first-served basis. Media Registration: Media may register on-site with their business cards and/or media identification. To view press releases in Chinese, please visit http://mediaroom.hktdc.com/tc The Hong Kong Trade Development Council (HKTDC) celebrates its 50th anniversary this year. The HKTDC is the international marketing arm for Hong Kong-based traders, manufacturers and services providers. With more than 40 offices globally, including 13 on the Chinese mainland, the HKTDC promotes Hong Kong as a platform for doing business with China and throughout Asia. The HKTDC also organises international exhibitions, conferences and business missions to provide companies, particularly SMEs, with business opportunities on the mainland and in overseas markets, while providing information via trade publications, research reports and digital channels including the media room. For more information, please visit: www.hktdc.com/aboutus. Follow us on Google+, Twitter @hktdc, LinkedIn.


News Article | April 6, 2016
Site: www.nature.com

C57BL/6J mice were purchased from Japan SLC (Shizuoka), and ICR mice and germ-free (IQI/Jic[Gf] ICR) mice were from CLEA Japan (Tokyo). Myd88−/− and Asc−/− mice backcrossed onto C57BL/6 mice for eight or more generations were used19, 20, 21. All mice were kept under SPF conditions at the Experimental Animal Facility, Graduate School of Medicine, Osaka University. All animal experiments were performed according to guidelines of the Animal Research Committee of the Graduate School of Medicine at Osaka University. RNA samples were prepared from indicated tissues using TRIzol reagent (Invitrogen). Total RNA was reverse transcribed using Moloney murine leukaemia virus reverse transcriptase (Promega) and random primers (Toyobo) after treatment with RQ1 DNase I (Promega). cDNA was analysed by real-time RT–PCR using GoTaq qPCR Master Mix (Promega) in ABI 7300 (Applied Biosystems). Values were normalized to the expression of Gapdh, and the fold difference in expression relative to that of Gapdh is shown. The following primer sets were used: Lypd8, 5′-GCCTTCACTGTCCATCTATTT-3′ and 5′-GTGACCATAGCAAGACATGCA-3′; Villin, 5′-CTATGCAGATGGTACCTGTTC-3′ and 5′-CCTGGGACGAGTCCTGGCCAA-3′; Muc2, 5′-ACATCACCTGTCCCGACTTC-3′ and 5′-GAGCAAGGGACTCTGGTCTG-3′; Lgr5, 5′-CATCACACTGTCACTGTGAGC-3′ and 5′-GGTAGCTGACTGATGTTGTTC-3′; Tnfa, 5′-TCCAGGCGGTGCCTATGT-3′ and 5′-CACCCCGAAGTTCAGTAGACAGA-3′; Il1b, 5′-TCAGGCAGGCAGTATCACTCA-3′ and 5′-GGAAGGTCCACGGGAAAGAC-3′; Ifng, 5′-TCAAGTGGCATAGATGTGGAAGAA-3′ and 5′-TGGCTCTGCAGGATTTTCATG-3′; Il6, 5′-CTGCAAGAGACTTCCATCCAGTT-3′ and 5′-AAGTAGGGAAGGCCGTGGTT-3′; Cx3cl1, 5′-GGCCGCGTTCTTCCATTTGT-3′ and 5′-TGATAGCGGATGAGCAAAGC-3′; Cxcl2, 5′-CTCAGTGCTGCACTGGTCCTG-3′ and 5′-CTGGGGGCGTCACACTCAAGC-3′; Ccl17, 5′-CTGCAGCATGCCAGAGCT-3′ and 5′-GGTCTTATACCAGCTCAC-3′; Ccl28, 5′-CAGCCTCACCTGAGTCATTGC-3′ and 5′-CAGTGCAACAGCTGGAGGCCA-3′; Gapdh, 5′-CCTCGTCCCGTAGACAAAATG-3′ and 5′-TCTCCACTTTGCCACTGCAA-3′. Plasmids (pCRII; Invitrogen) containing a cDNA fragment of Lypd8 were used as templates of RNA probes. Digoxigenin (DIG)-labelled antisense or sense probes were prepared with T7 and SP6 RNA polymerase (Ambion), respectively, using the DIG RNA Labelling Mix (Roche Diagnostics). Colonic tissues of C57BL/6J were fixed with 4% paraformaldehyde (PFA). Serial frozen sections were fixed in 4% PFA for 20 min, incubated in cold 0.1% H O , and permeabilized with 50 μg ml−1 proteinase K for 5 min. After an additional fixation with PFA, the sections were treated with acetic anhydrite in triethanolamine for 5 min. The sections were then pre-hybridized with 50% formamide, 5 × saline sodium citrate, 1 mg ml−1 yeast tRNA (Roche Diagnostics), 100 μg ml−1 heparin, 1 × Denhardt’s solution, 0.1% Tween 20 at 60 °C for 3 h, and hybridized at 60 °C for 16 h. After washing, sections were incubated with horseradish peroxide (HRP)-anti-DIG Fab fragment (clone 1.71.256: Roche Diagnostics), followed by biotin-labelled tyramide (TSA Biotin System; Perkin Elmer) for signal amplification. Hybridized probes were detected by ABC-Alkaline phosphatase (Vector Laboratories) and NBT/BCIP (Roche Diagnostics). Images were obtained using BZ-9000 (Keyence). Targeting vectors were constructed by replacement of genomic fragment containing the third and fourth exons of Lypd8 with a Cre recombinase internal ribosome entry site Venus neomycin-resistance gene cassette (Lypd8venus mice) or a neomycin-resistance gene cassette (Lypd8−/− mice), and a gene encoding HSV thymidine kinase driven by a phosphoglycerate kinase promoter inserted into the genomic fragment. After the targeting vector was transfected into V6.5 embryonic stem cells, G418 and ganciclovir double-resistant colonies were selected and screened by PCR and Southern blot analysis. Homologous recombinants were used for generation of Lypd8venus mice and Lypd8−/− mice. Lypd8venus mice and Lypd8−/− mice were backcrossed onto C57BL/6 mice for at least six generations, and Lypd8venus mice, Lypd8−/− mice and their wild-type littermates from intercrosses of heterozygous mice were kept in the same cages and used for experiments. There was no randomization, but stratification was used to achieve the similar ages and sex rations among experimental groups. The experiments were not blinded. Flow cytometry of isolated colonic epithelial cells Intestinal epithelial cells were isolated from Lypd8venus mice by shaking intestinal tissues in 5 mM EDTA/HBSS solution at 37 °C for 20 min and then washed with phosphate buffered saline (PBS). The intestinal epithelial cells were treated with PE-Cy7-conjugated anti-CD3ε Ab (clone 145-2C11: BD Biosciences) in PBS containing 2% FBS to block nonspecific binding. Flow cytometric analysis was performed using a FACSCanto II flow cytometer (BD Biosciences) with FlowJo software (Tree Star). For cell isolation, cells were sorted using a FACSAria (BD Biosciences). A sequence for Flag-tagged Lypd8 was constructed by inserting Flag-tag sequence into total Lypd8 coding sequence immediately downstream of predicted N-terminal signal sequence. CMT-93 cells and Caco-2 cells, which were originally obtained from ATCC and free of mycoplasma, were transfected with linearized pcDNA3.1 (+) vector (Invitrogen) inserted the sequence for Flag-tagged Lypd8 using Lipofectamine2000 (Invitrogen). These cells were cultured in G418-containing medium. The surviving cells were stained with anti-Flag M2 monoclonal antibody (cat F3165: Sigma-Aldrich) and Alexa Fluor 488 goat anti-mouse IgG antibody (cat A11001: Molecular Probes) and cells expressing Flag-tagged Lypd8 were sorted using FACSAria (BD Biosciences). Caco-2 cells stably expressing Flag-tagged Lypd8 (1 × 105 cells) were cultured on transwell filters of 3.0-μm pore size (BD Biosciences) for 2 weeks. After confirming full confluency, Caco-2 cells were washed with PBS, fixed with 4% PFA and then blocked by 1% bovine serum albumin (BSA) in PBS. Cells were stained with mouse anti-Flag M2 mAb (Sigma-Aldrich) plus Alexa Fluor 488 goat anti-mouse IgG (Invitrogen) or rabbit anti-human Claudin-1 Ab (cat ab15098; Abcam) plus Alexa Fluor 594 goat anti-rabbit IgG (cat A11012; Invitrogen), and counterstained with DAPI (Vector Laboratories). Transwell filters with these cells were cut, placed on slide glasses and analysed using a confocal microscope (FV1000-D; Olympus). The supernatants on Caco-2 cells with or without Lypd8 expression were incubated with anti-Flag M2 affinity gel (Sigma-Aldrich) at 4 °C for 3 h. The resin was collected, washed three times with Tris-buffered saline and then suspended in sample buffer for immunoblot analysis using anti-Flag M2 mAb (Sigma-Aldrich) and HRP-conjugated goat anti-mouse IgG (cat NA931: GE Healthcare). CMT93 cells stably expressing Lypd8 (1 × 106 cells) were rinsed twice with cold PBS and incubated with 0.5 ml of the same buffer containing 0.5 units of Bacillus cereus phosphatidylinositol-specific phospholipase C (Invitrogen) at 4 °C for 20 min. These cells were stained with anti-Flag M2 mAb (Sigma-Aldrich) and Alexa Fluor 488 goat anti-mouse IgG (Invitrogen). The surface expression of Lypd8 was analysed using a FACSCanto II (BD Biosciences). Lypd8−/− mice were immunized with P3U1 cells with retroviral overexpression of Lypd8. Lymph node cells from immunized Lypd8−/− mice were fused with Sp2/0 mouse myeloma cells. Hybridomas were screened by analysing reactivity to RBL1 cells overexpressing Flag-tagged Lypd8 with flow cytometry. Positive clones were selected for further subcloning. Culture supernatants of subcloned hybridoma cells were screened by immunostaining of 4% PFA- or Carnoy’s-fixed colon sections from wild-type and Lypd8−/− mice. Culture supernatants of hybridoma clones (clone number 4F8 and 5A4), which stained Carnoy’s-fixed colon sections of wild-type mice, but not Lypd8−/− mice, were used as mouse anti-mouse Lypd8 mAb. The specificity of mAb was also confirmed by flow cytometry analysis of CMT93 cells with or without Lypd8 expression. Anti-mouse Lypd8 mAb was purified from the supernatant by Ex-pure Spin ProG (Kyoto monotech) and labelled with CF633 using Mix-n-Stain CF633 antibody labelling kit (Biotium). Colons from 8- to 12-week-old mice without washing were fixed in methanol–Carnoy’s fixative composed of 60% methanol, 30% chloroform and 10% acetic acid. Paraffin-embedded sections were dewaxed and hydrated. Sections were blocked with 1% BSA in PBS and stained with CF633-conjugated anti-mouse Lypd8 mAb or anti-Mucin2 Ab (clone H-300: Santa Cruz Biotechnology) and Alexa Fluor 594 goat anti-rabbit IgG (cat A11012: Invitrogen). Sections were incubated with 1 μg Cy3- or Cy5-conjugated EUB338 (5′-GCTGCCTCCCGTAGGAGT-3′) for detection of all bacteria or Cy3-conjugated pB-02110 (5′-ATGGGTTCATCCCATAGTGC-3′)14 for detection of Proteus spp. in 200 μl of hybridization buffer (750 mM NaCl, 100 mM Tris-HCl (pH 7.4), 5 mM EDTA, 0.01% BSA, 10% dextran sulfate) at 40 °C for 16 h. Sections were rinsed in wash buffer (50 mM NaCl, 4 mM Tris-HCl (pH 7.4), 0.02 mM EDTA), washed at 45 °C for 20 min and counterstained with DAPI (Vector Laboratories). Human colons from the normal mucosa of resected colon tissues from colon cancer patients with no history of a diagnosis of inflammatory bowel diseases or the inflamed mucosa of resected colon tissues from ulcerative colitis patients were fixed in 4% PFA. The experiment was approved by the Ethical Committee of Osaka University School of Medicine. All participants provided informed consent. All ulcerative patients had a confirmed diagnosis by a gastroenterologist. Paraffin-embedded sections were dewaxed and hydrated. Sections were blocked with 1% BSA in PBS and stained with anti-human Lypd8 Ab (clone V-16: Santa Cruz Biotechnology) and Alexa Fluor 568 donkey anti-goat IgG (cat A11057: Invitrogen). Normal goat IgG (cat AB-108-C: R&D systems) was used as an isotype control. Sections were analysed using a confocal microscope (FV1000-D; Olympus). The distance between bacterial populations and the epithelial surface was measured at four points of the proximal, middle and distal colon in each mouse. Recombinant Lypd8 or LOC69864 (an uncharacterized protein that is structurally most similar to Lypd8) was purified from CMT93 cells stably expressing FLAG-tagged Lypd8 or LOC69864 using FLAG M Purification Kit (Sigma-Aldrich). As a negative control, cells transfected with empty vector (pcDNA3.1 (+) (Invitrogen)) were used. Recombinant Lypd8 proteins (1 μg) were incubated with PNGase F, sialidase A and O-glycanase (Prozyme) at 37 °C for 3 h. Recombinant Lypd8 proteins treated with glycanase were separated with SDS–PAGE and transferred to polyvinylidene fluoride membranes (Millipore) that were incubated with anti-Flag M2 mAb (Sigma-Aldrich) and then HRP-conjugated goat anti-mouse IgG (cat NA931: GE Healthcare). Immunoreactivity was detected using SuperSignal (Thermo Scientific). The mutant Lypd8 N–D sequence was designed so that thirteen Asp (N) residues were converted to Asn (D). 293T cells were transfected with the mutant Lypd8 (N–D) expression vector using Lipofectamine 2000 (Invitrogen) and cell lysates of 293T cells expressing mutant Lypd8 (N–D) protein were separated with SDS–PAGE and analysed by western blot. Faeces, luminal contents of the colon or colonic tissues were collected in tubes containing RNAlater (Ambion). After weights were measured, RNAlater was added to make tenfold dilutions of homogenates. Homogenates (200 μl) of faeces, luminal contents or colonic tissues, or bacterial suspension containing sorted bacteria were washed twice with 1 ml PBS, 0.3 g glass beads (diameter, 0.1 mm) (BioSpec Products), 300 μl Tris-SDS solution and 500 μl TE-saturated phenol were added to the suspension, and the mixture was vortexed vigorously using a FastPrep-24 (M.P. Biomedicals) at 5.0 power level for 30 s. After centrifugation at 20,000 g for 5 min at 4 °C, 400 μl of supernatants were collected. Subsequently, phenol–chloroform extraction was performed and 250 μl of supernatants were subjected to isopropanol precipitation. Finally, DNAs were suspended in 200 μl TE buffer and stored at −20 °C. PCR was performed using a primer set (784F, 5′-AGGATTAGATACCCTGGTA-3′; and 1061R, 5′-CRRCACGAGCTGACGAC-3′) targeting the V5–V6 region of the 16S rRNA genes with KAPA HiFi HotStart Ready Mix (KAPA Biosystems). Products were purified using DNA clean and Concentrator-5 (Zymo Research). Adaptor and barcode sequences were attached to the products by 10 cycles of PCR with 1 ng of each of initial PCR product as the template and primer sets and other PCR conditions were unchanged. Sequencing was performed using a 316 chip and Ion PGM Sequencing 400 Kit (Life Technologies) on the Ion PGM sequencer (Life Technologies). Raw sequences were demultiplexed and quality-trimmed by the following procedures: (1) trimming bases with quality below Q15 from 3′ end of each read, (2) removing reads with average quality below Q20, (3) removing reads without primer sequences on both ends, and (4) removing reads with length shorter than 260 basepairs, using the FASTX-Toolkit (http://hannonlab.cshl.edu/fastx_toolkit/index.html) and BBtrim (http://bbmap.sourceforge.net/). The processed sequences were then clustered into operational taxonomic units (OTU) defined at 94% similarity cutoff using UCLUST version 1.2.22q. Representative sequences for each OTU were classified taxonomically using RDP Classifier version 2.2 with the SILVA 111 database. Quantitative PCR was performed in ABI7300 using GoTaq qPCR Master Mix (Promega). The following primer sets were used: ‘all bacteria’, 5′-CGGTGAATACGTTCCCGG-3′ and 5′-TACGGCTACCTTGTTACGACTT-3′; Bacteroides, 5′-GAGAGGAAGGTCCCCCAC-3′ and 5′-CGCTACTTGGCTGGTTCAG-3′; Prevotella, 5′-CACRGTAAACGATGGATGCC-3′ and 5′-GGTCGGGTTGCAGACC-3′; Lactobacillus, 5′-TGGAAACAGRTGCTAATACCG-3′ and 5′-GTCCATTGTGGAAGATTCCC-3′; Bifidobacterium, 5′-CTCCTGGAAACGGGTGG-3′ and 5′-GGTGTTCTTCCCGATATCTACA-3′; Escherichia/Shigella, 5′-GAGTAAAGTTAATACCTTTGCTCATTG-3′ and 5′-GAGACTCAAGCTKRCCAGTATCAG-3′; Helicobacter, 5′-CTATGACGGGTATCCGGCC-3′ and 5′-TCGCCTTCGCAATGAGTATT-3′; Staphylococcus, 5′-TTTGGGCTACACACGTGCTACAATGGACAA-3′ and 5′-AACAACTTTATGGGATTTGCWTGA-3′, Enterococcus, 5′-ATCAGAGGGGGATAACACTT-3′ and 5′-ACTCTCATCCTTGTTCTTCTC-3′; and Proteus, 5′-GTTATTCGTGATGGTATGGG-3′ and 5′-ATAAAGGTGGTTACGCCAGA-3′. In the analysis of Lypd8-bound and unbound bacteria, all samples were normalized to the 16S rRNA gene level of ‘all bacteria’. Colonic tissues were thoroughly washed twice and collected in tubes containing 5 ml PBS. After colonic tissues were homogenized in PBS using a homogenizer, 100 μl of the homogenates were incubated on lysogeny broth (LB) or MacConkey agar plates at 37 °C in Ruskinn Bugbox Plus anaerobic chamber (The Baker Company) for 24 h. The number of swarming colonies on LB agar plates or white colonies on MacConkey agar plates was counted at 16 h after the start of incubation and colony-forming units were calculated. P. mirabilis was cultured in LB medium at 37 °C until OD reached 0.6. The bacterial culture was centrifuged at 8,000g for 5 min and the pellet was resuspended in 1 ml PBS. The bacterial suspension was re-centrifuged and the pellet was labelled with CFSE Fluorescent Cell Labelling Kit (Abcam). After confirming CFSE labelling of P. mirabilis using a FACSCanto II (BD Biosciences), CFSE-labelled P. mirabilis was suspended with PBS (5 × 108 per ml) and the bacterial suspension (200 μl) was inoculated into anaesthetized mice via the transanal route with a catheter. At 1, 2, 4, and 8 h after inoculation, colons without washing were fixed in 4% PFA. Colon sections were counterstained with DAPI (Vector Laboratories), and analysed using a confocal microscope (FV1000-D; Olympus). Eight- to ten-week-old Lypd8−/− mice and their littermate wild-type mice were used for DSS-induced colitis experiments. Acute colitis was induced by administration of 2% DSS (36–50 kDa; MP Biomedicals) in the drinking water for 5 days. Mice were analysed for changes in weight, survival rates and histological changes. Mice were treated with gentamicin (2 g l−1; Nacalai Tesque) or vancomycin (500 mg l−1; Duchefa Biochemie B.V.) dissolved in autoclaved drinking water and provided for 2 weeks. Fluid intake was monitored. Faeces were collected in tubes containing PBS. After weights were measured, PBS was added to make tenfold diluted suspensions. PBS-diluted faeces were mixed well and centrifuged at 400g for 5 min to remove larger particles from bacteria. Supernatants (200 μl) were centrifuged at 8,000g for 10 min to remove non-bound immunoglobulins. Pellets were resuspended in 1 ml PBS containing 2% FBS, and used as a bacterial suspension. Bacterial suspensions from wild-type mice and Lypd8−/− mice were incubated with anti-Lypd8 antibody. After washing with PBS containing 2% FBS, bacterial pellets were stained with Alexa Fluor 647 goat anti-mouse IgG1 (Invitrogen). Bacterial suspensions from C57BL/6J mice were incubated with recombinant Lypd8 protein (1 μg) on ice for 30 min and washed with PBS. Next, bacterial pellets were treated with anti-Flag M2 mAb (Sigma-Aldrich). Finally, bacterial pellets were stained with Alexa Fluor 488 goat anti-mouse IgG (Invitrogen). Bacterial suspensions were analysed using a FACSCanto II (BD Biosciences) with FlowJo software (Tree Star) or a confocal microscope (FV1000-D; Olympus). Lypd8-bound and unbound bacteria were sorted using a FACSAria (BD Biosciences). Bacterial strains, Bacteroides sartrii JCM 17136T, Lactobacillus acidophilus JCM 1132T, Bifidobacterium breve JCM 1192T and Enterococcus gallinarum JCM 8728T, were obtained from Japan Collection of Microorganisms. P. mirabilis was isolated from the colonic tissue of Lypd8−/− mice. All bacteria were cultured in Gifu anaerobic medium (Nissui) under anaerobic conditions. The pure preparation of each bacteria was centrifuged at 8,000g for 5 min and the pellet was resuspended in 1 ml PBS containing 2% FBS. Bacteria were incubated with recombinant Lypd8 protein (1 μg) on ice for 1 h and washed with PBS. Next, the bacterial pellet was treated with anti-Flag M2 mAb (Sigma-Aldrich). Finally, the bacterial pellet was stained with Alexa Fluor 488 goat anti-mouse IgG (Invitrogen) and analysed using a FACSCanto II (BD Biosciences). Bacterial suspension was mixed with FLAG-tagged recombinant Lypd8 (rLypd8; 20 ng μl−1), and then incubated for 1 h at 4 °C. After the incubation, bacterial suspensions were centrifuged, and pellet was resuspended in PBS. A drop of bacterial suspension was placed on an 3-aminopropyltriethoxysilane (APS)-coated cover slip (Matsunami Glass IND), and blocked with bovine serum albumin and normal goat serum. The mounted bacteria were reacted with anti-FLAG M2 mAb (Sigma-Aldrich), washed with PBS, and reacted with 5-nm-gold-labelled anti-mouse IgG goat antibody (EY Laboratories). The sample was fixed with 2% glutaraldehyde and 1% OsO , dehydrated in a graded ethanol series and 3-methylbutyl acetate, and dried in a critical-point drying chamber (HCP-1, Hitachi High-Technologies). It was then coated with a platinum layer approximately 1 nm thick in an ion sputter coater (E-1030, Hitachi High-Technologies), and examined under a scanning electron microscope (S-5000, Hitachi High-Technologies). Bacterial suspension of P. mirabilis or E. coli JCM 1649T (Japan Collection of Microorganisms) in PBS was shaken 300 times per min for 60 min to remove flagella from bacterial bodies. Bacterial bodies were pelleted by centrifuging at 4,000g for 20 min. The supernatants were ultracentrifuged at 80,000g for 60 min to obtain flagella. Bacterial bodies or flagella were mixed with the solution of FLAG-tagged recombinant Lypd8 (10 ng μl−1) or LOC69864 (10 ng μl−1), and incubated for 3 h at 4 °C. After the incubation, the bacterial bodies or flagella were pelleted by centrifugation or ultracentrifugation, respectively. Then, the supernatant was collected and the pellet was resuspended in PBS. The supernatant and pellet suspension were separated with SDS–PAGE and transferred to polyvinylidene fluoride membranes (Millipore) that were incubated with anti-Flag M2 mAb (Sigma-Aldrich) and then HRP-conjugated goat anti-mouse IgG (cat NA931: GE Healthcare). Immunoreactivity was detected using SuperSignal (Thermo Scientific). Bacterial DNA of P. mirabilis was extracted from the culture of P. mirabilis using a previously described protocol (see above). Using the extracted bacterial DNA as template, PCR was performed to isolate the flagellin gene with a primer set (5′-GGATCCGCACAAGTTATTAATACTAATTAT-3′ and 5′-GCGGCCGCTTAACGTAACAGAGACAGAACAGT-3′) targeting the full-length flagellin gene of P. mirabilis. The isolated DNA was cloned in frame into EcoRI- and NotI-digested pGEX-6P-2 (GE Healthcare) to generate a glutathione S-transferase (GST)–flagellin fusion construct. A recombinant flagellin protein was prepared by growing E. coli harbouring the pGEX-flagellin plasmids in LB medium containing 100 μg ml−1 of ampicillin at 37 °C until OD reached 0.6. After growth, 1 mM isopropyl β-d-1-thiogalactopyranoside (IPTG) was added to the medium, and bacterial culture was continued for 4 h at 37 °C. Cells were centrifuged and the cell pellet was suspended in PBS with 1% Triton X-100. The cell suspension was sonicated 5 times for 30 s at 4 °C, and cleared by centrifugation. The resulting solution was bound to Glutathione-Sepharose 4 Fast Flow (GE Healthcare). The beads were washed 3 times. The GST fusion proteins were then eluted with elution buffer (5 mM glutathione, 50 mM Tris-HCl, pH 9.6). Elution buffer was replaced with PBS with Amicon Ultra-15 (Millipore). The amounts and purity of the protein were estimated by SDS–PAGE. 96-well plates (Corning) were coated with 5 μg ml−1 GST peptides, 5 μg ml−1 GST-tagged flagellin protein of P. mirabilis, 100 μg ml−1 flagella of P. mirabilis or PBS 1% BSA for 16 h at 4 °C. Plates were then washed, and increasing concentrations of mouse TLR5 Fc protein (R&D systems) or FLAG-tagged Lypd8 protein diluted in PBS 1% BSA were added and incubated for 2 h at room temperature. Plates were then washed, and 0.5 μg ml−1 of HRP-conjugated goat anti-mouse IgG (cat NA931: GE Healthcare) or HRP-conjugated anti-FLAG M2 mAb (Sigma-Aldrich) diluted in PBS 1% BSA was added and incubated for 2 h at room temperature. Plates were then washed, and TMB substrate was added. Plates were then read at 450 nm with a spectrometer. P. mirabilis (1 × 105) were incubated in LB medium (1 ml) with the indicated concentrations of recombinant Lypd8 protein for 6 h at 37 °C. The bacterial cultures were applied to MacConkey agar plates and incubated at 37 °C for 16 h, and the number of colonies was counted and colony-forming units were calculated. Human colon epithelia were obtained from the normal mucosa of resected colon tissues from colon cancer patients. Colonic epithelial layers were first stripped by shaking sections of colon in 5 mM EDTA/HBSS. Total RNA was isolated from Caco-2 cells and human colon epithelia and reverse transcribed. cDNA was analysed by real-time RT–PCR. Values were normalized to the expression of GAPDH, and the fold difference in expression relative to that of GAPDH is shown. The following primer sets were used: LYPD8, 5′-GAACACTTTCATTTTGTAAGC-3′ and 5′-ACGACAGGAAGTTCCATTAGA-3′; GAPDH, 5′-TGGATATTGTTGCCATCAATG-3′ and 5′-TGATGGGATTTCCATTGATGA-3′. The experiment was approved by the Ethical Committee of Osaka University School of Medicine and informed consent for specimen use was obtained from all patients. E. gallinarum or P. mirabilis (1 × 105) was added to a culture of polarized Caco-2 cells. For PNGase F treatment, cells were irradiated with 30 Gy to stop cell growth and then incubated in culture medium containing PNGaseF (Sigma-Aldrich) (2 unit/ml) for 2 h. At 8 h after the addition of P. mirabilis, each well was washed with PBS three times and then Caco-2 cells were removed from transwell filters with trypsin/EDTA solutions and suspended in PBS. PBS containing Caco-2 cells were applied to LB or MacConkey agar plates and incubated at 37 °C for 16 h, and the number of colonies was counted and colony-forming units were calculated. P. mirabilis was cultured in LB medium at 37 °C until OD was 0.6. LB agar plates (1.5%) were centrally inoculated with 5 μl of bacterial culture and incubated at 37 °C. After 1 h, polarized Caco-2 cells with or without Flag-tagged Lypd8 expression, cultured on 75 mm diameter transwell filters of 3.0 μm pore size (BD Biosciences) for approximately 2 weeks, were added to P. mirabilis-inoculated agar plate so that the apical side of cells adhered to the surface of the agar plate. Motility was assessed by examining circular swarms formed by growing motile bacterial cells. The length from the centre to four points of the edge of circular swarms was measured at 5, 9 and 13 h time points following P. mirabilis inoculation. CMT-93 cells (5 × 107 cells) with or without Flag-tagged Lypd8 were lysed with 1 ml of CelLytic M Cell Lysis Reagent (Sigma-Aldrich). After removal of any insoluble material by centrifugation and filtration with a 0.45 μm filter, the cell lysates were incubated with 50 μl of anti-Flag M2 affinity gel (Sigma-Aldrich) for 3 h. The resin was centrifuged and washed with Tris-buffered saline three times. The resin containing Flag-tagged Lypd8 was mixed with 1 ml of LB medium containing 0.3% agar. The final concentration of Lypd8 in LB agar was estimated by SDS–PAGE and was approximately 1.25 μg ml−1. P. mirabilis and E. coli were cultured in LB medium at 37 °C until OD was 0.6. Semisolid LB agar (0.3%) containing Flag-tagged Lypd8 were centrally inoculated with 0.5 μl of bacterial culture and incubated at 37 °C. Motility was assessed by examining circular migration. The radius of circles formed by bacterial migration was measured at 4 h after the bacterial inoculation. Data are presented as mean ± s.d., as indicated in the figure legends. Differences between control and experimental groups were evaluated using a two-tailed unpaired Student’s t-test. For the swarming motility assay, a paired Student’s t-test was performed. A P value of <0.05 was considered significant. For the determination of microbiota by deep sequencing, principal component analysis was used to visualize data sets by statistical programming language R 2.1.5. No statistical methods were used to predetermine sample size. No animal or sample was excluded from the analysis.


News Article | December 15, 2016
Site: www.eurekalert.org

NEW YORK NY (December 15, 2016)--Gastric tumors are started by specialized cells in the stomach that signal nerves to make more acetylcholine, according to a study in mice. The multinational team of researchers who conducted the study also identified a substance called nerve growth factor that stimulates nerve development and, when blocked, inhibits stomach cancer development. The findings were published today in Cancer Cell. Previous studies have shown that nerves are abundant in the gastric tumor microenvironment. In an earlier paper, the researchers demonstrated that inhibiting signaling by the neurotransmitter acetylcholine, by severing the vagus nerve in the stomach or treating with Botulinum toxin, shrank or prevented the growth of gastric tumors in mouse models. "Nerves and acetylcholine clearly play a key role in regulating the development and growth of cancer cells, particularly cancer stem cells, in the gastric tumor microenvironment," said Timothy C. Wang, MD, the Dorothy L. and Daniel H. Silberberg Professor of Medicine at Columbia University Medical Center (CUMC) and senior author of the paper. "But little is known about what is driving cancer in the earliest stage of development, before the expansion of nerves in the microenvironment. We also wanted to find out where acetylcholine is coming from before the growth of nerves." Through a series of experiments in mouse models, the researchers determined that a neurotrophin (substance that triggers nerve growth) called nerve growth factor is highly expressed in gastric cancer cells. They also discovered that tuft cells--specialized cells found in the lining of the digestive tract that, like nerves, communicate with other cells--provide another source of acetylcholine for cancer cell growth, particularly during the formation of tumors. "We learned that tuft cells are increased during the earliest stage of gastric tumor development, making acetylcholine and stimulating the production of nerve growth factor within the lining of the stomach," said Dr. Wang. "As nerves grow in around the tumor, tuft cells decrease." In additional experiments, the scientists showed that overexpression of nerve growth factor in the mouse stomach drove tumorigenesis. Furthermore, administration of a nerve growth factor receptor inhibitor prevented stomach cancer in the mice. "Our study provides some insight into the cellular crosstalk that leads to the development of stomach cancer, and points to a viable therapeutic target for this type of cancer," said Dr. Wang. "Using our findings as a paradigm, additional studies can be done to identify the specific neurotrophins and neurotransmitters that are involved in tumor development in other areas of the body." The study is titled, "Nerve growth factor promotes gastric tumorigenesis through aberrant cholinergic signaling." The other contributors are: Yoku Hayakawa (University of Tokyo, Tokyo, Japan), Kosuke Sakitani (University of Tokyo), Mitsuru Konishi (University of Tokyo), Samuel Asfaha (University of Western Ontario, Ontario, Canada), Ryota Niikura (University of Tokyo), Hiroyuki Tomita (Gifu University Graduate School of Medicine, Gifu, Japan), Bernhard W. Renz (Hospital of the University of Munich, Munich, Germany), Yagnesh Taylor (CUMC), Marina Macchini (CUMC). Moritz Middlehoff (CUMC), Zhengyu Jiang (CUMC), Takayuki Tenaka (CUMC), Zinaida A. Dubeykovskaya (CUMC), Woosook Kim (CUMC), Xiaowei Chen (CUMC), Aleksandra M. Urbanska (CUMC), Karan Nagar (CUMC), Christoph B. Westphalen (Klinikum der Universität München, Munich, Germany), Michael Quante (Technische Universität München, Munich, Germany), Chyuan-Sheng Lin (CUMC), Michael D. Gershon (CUMC), Akira Hara (Gifu University Graduate School of Medicine), Chun-Mei Zhao (Norwegian University of Science and Technology, Trondheim. Norway), Duan Chen (Norwegian University of Science and Technology), Daniel L. Worthley (University of Aidelaide, Australia), and Kazuhiko Koike (University of Tokyo). The study was supported by grants from the National Institutes of Health (U54CA126513, R01CA093405, R01CA120979, and R01DK052778), the Clyde Wu Family Foundation, the Nakayama Cancer Research Institute, the Okinaka Memorial Institute for Medical Research, and the Project for Cancer Research and Therapeutic Evolution from the Japan Agency of Medical Research and Development. Y.H. and K.S. were supported by Japan Society for the Promotion of Science, and Y.H. and T.T. were supported by Uehara Memorial Foundation. The authors declare no conflicts of interest. Columbia University Medical Center provides international leadership in basic, preclinical, and clinical research; medical and health sciences education; and patient care. The medical center trains future leaders and includes the dedicated work of many physicians, scientists, public health professionals, dentists, and nurses at the College of Physicians and Surgeons, the Mailman School of Public Health, the College of Dental Medicine, the School of Nursing, the biomedical departments of the Graduate School of Arts and Sciences, and allied research centers and institutions. Columbia University Medical Center is home to the largest medical research enterprise in New York City and State and one of the largest faculty medical practices in the Northeast. The campus that Columbia University Medical Center shares with its hospital partner, NewYork-Presbyterian, is now called the Columbia University Irving Medical Center. For more information, visit cumc.columbia.edu or columbiadoctors.org.


TOKYO--(BUSINESS WIRE)--ON Semiconductor (Nasdaq: ON), driving energy efficient innovations, announced that Osamu Takiguchi joins the company as president and representative director of ON Semiconductor Japan, Ltd. to lead and manage the sales and marketing operations in Japan. Takiguchi is also concurrently serving as the corporate vice president of Japan regional sales in order to expand its Japan business and align with corporate strategies in key markets. “Takiguchi brings more than 30 years of semiconductor industry experience to our company in design engineering and sales. He is effective in engaging in the global marketplace and will further strengthen our footprint in Japan with our valued customers,” said Paul Rolls, executive vice president sales and marketing for ON Semiconductor. “Takiguchi will be integral to our strategy in Japan and will further drive total solutions in our focus markets of automotive and industrial.” The company has invested in the Japan market and the investment in Japan has played an important role in driving the company’s global strategy focusing on automotive, industrial, IoT and power conversion and motor control segments. The investment includes design centers for product development, manufacturing facilities, a solution engineering center and fault analysis capability in Japan to establish a reliable environment for its Japanese customers base. “I look forward to strengthening the partnerships with our customers by continuing to demonstrate our value-add in the Japanese market,” said Takiguchi. “We have a strong, broad product portfolio and are a unique company in the country offering full solutions to our customers. Our strong customer service and responsive supply chain stand out in our industry.” ON Semiconductor offers more than 64,000 products and leading-edge system solutions. The company’s focused end-markets within Japan include automotive, industrial, white goods and other consumer products. Driving expanded adoption of ON Semiconductor system solutions, custom ASIC capabilities, mixed-signal, analog and industry-leading standard products into all electronics end-markets is a key part of the company’s Japan customer support strategy. The company has had operations based in Japan since 1999, when ON Semiconductor spun-off from Motorola Inc., and at that time took on manufacturing facilities within the country that dated back to the mid-1980s. Among the company’s Japan-based operations are sales offices and a Solutions Engineering Center (SEC) dedicated to supporting customers’ product development needs, two Design Centers located in Gunma and Gifu and both 5-inch and 6-inch wafer manufacturing facilities at its Niigata campus. A listing of ON Semiconductor’s Japan sales office and distribution partners can be found online via this LINK. ON Semiconductor (Nasdaq: ON) is driving energy efficient innovations, empowering customers to reduce global energy use. The company is a leading supplier of semiconductor-based solutions, offering a comprehensive portfolio of energy efficient power management, analog, sensors, logic, timing, connectivity, discrete, SoC and custom devices. The company’s products help engineers solve their unique design challenges in automotive, communications, computing, consumer, industrial, medical, aerospace and defense applications. ON Semiconductor operates a responsive, reliable, world-class supply chain and quality program, a robust compliance and ethics program, and a network of manufacturing facilities, sales offices and design centers in key markets throughout North America, Europe and the Asia Pacific regions. For more information, visit http://www.onsemi.com. ON Semiconductor and the ON Semiconductor logo are registered trademarks of Semiconductor Components Industries, LLC. All other brand and product names appearing in this document are registered trademarks or trademarks of their respective holders. Although the company references its Web site in this news release, such information on the Web site is not to be incorporated herein.


TOKYO, Dec. 27, 2016 /PRNewswire/ -- VT HOLDINGS (TOKYO: 7593) is pleased to announce the results for its 2016 second quarter. Although Nissan Motor Company suspended sales of two vehicle models from April to June due to the vehicle supplier fuel economy test data scandal, vehicle sales edged up 1.3% compared to the previous fiscal year. As a result, consolidated net sales were JPY 73,469 million (+1.9% YoY), operating income was JPY 3,042 million (-19.2% YoY), ordinary income was JPY 3,046 million (-18.6% YoY) and profit attributable to owners of parent was JPY 1,690 million (-10.2% YoY). In the automotive sales-related business, net sales were JPY 71,072 million (+2.7% YoY) and operating income was JPY 3,205 million (-17.2% YoY). In this segment, Honda vehicle sales declined 2.7% and Nissan vehicle sales declined 19.7% due to the suspended sales of two vehicle models. However, two new consolidated subsidiaries were added to the Group, one in Japan and the other overseas, causing overall Group new vehicle sales, including those overseas, to increase 0.7% compared to the same period in the previous fiscal year. In terms of used cars, this business struggled with only 2,940 vehicles exported overseas, a year-on-year decrease of 18.6%. However, vehicle exports for the Group overall increased 1.6% year-on-year to surpass the previous year on a per vehicle basis. At the same time, in the services category, the rental car business saw increases in sales and income. In the housing-related business, net sales were JPY 2,325 million (-16.3% YoY) and operating income was JPY 94 million (+49.0% YoY). This segment develops condominium sales businesses in Aichi and Gifu Prefectures, and detached home sales businesses with branch offices in Tokyo, Osaka and Nagoya. The condominium sales business performed well this quarter on sales of properties where construction has been completed. The transfer of Shizuoka BMW business and the inclusion of WESSEX GARAGES HOLDINGS LIMITED in the scope of consolidation resulted in an increase in current assets including merchandise and finished goods and work in progress, as well as an increase in buildings and structures and machinery, equipment and vehicles under non-current assets, resulting in total assets expanding to JPY 10.3 billion. On November 9, VT HOLDINGS announced upward revisions to its full-year forecast. The reasons for this include the October 3, 2016, acquisition of Master Automocion S. L., a vehicle dealer group headquartered in Spain, as a consolidated subsidiary. Having carefully examined the material impact of this acquisition, and in consideration of Nissan's suspended sales of two light vehicle models in April, which were resumed earlier than initially expected in July, as well as initial exchange rate assumptions that did not anticipate yen appreciation, the Company revised exchange rates to a more appropriate level. VT HOLDINGS CO., LTD. (7593, First Section, TSE) 'Summary of Consolidated Financial Results for the Six Months Ended September 30, 2016' are available. http://www.vt-holdings.co.jp/eng/ir/library/pdf/20161111_2Q.pdf This release is for the purpose of providing information to serve as a reference for investment decisions and not for the purpose of soliciting investment. Please make your own judgment on final decisions such as investment policy, timing and selection. Please be advised that we do not assume any responsibility for damages caused by this service. Borderless IR specializes in the overseas distribution of IR content, including the dissemination of newsletters and annual reports providing the latest information and main strengths of Japanese companies directly to overseas investors through leading global media, corporate information database services and mailing lists. Borderless is also engaged in supporting other global IR efforts. To view the original version on PR Newswire, visit:http://www.prnewswire.com/news-releases/vt-holdings-co-ltd-7593-first-section-tokyo-stock-exchange-issues-operating-performance-in-the-second-quarter-of-the-fiscal-year-ending-march-31-2017-300383588.html


TOKYO, Nov. 25, 2016 /PRNewswire/ -- HULS Inc., a company which specialises in the global business of Japanese craft industries based in Tokyo and Singapore, launches "Cloud" espresso cup and saucer on 15th December 2016 both in Singapore and in Japan. The product is a result of the collaboration between a Singaporean designer and a Japanese ceramic manufacturer in the Mino region. "Cloud" is a white porcelain espresso cup and saucer inspired by the closeness of the clouds and skies over Singapore. It is designed by a Singaporean designer, Casey Chen and manufactured by Miyama, a Japanese ceramic company located in Gifu prefecture which is renowned for their Mino ware and under the direction of HULS. Casey was inspired by dedication and attentiveness to details of the workers and the unique decoration technique of applying patterns on porcelain using Japanese paper which characterises Miyama's specialty in the field of Japanese ceramic tableware. The collaboration between Singapore and Japan, in the creative and craft industries had flourished in recent years. Singapore's unique cultural diversity and Japan's craft industries are both rooted in tradition and this combination became the source of the creation of the finished product. The blue detailing of the saucer symbolises both the sky as well as the Singapore Peranakan blue and the matte texture of bisque porcelain realises a quality feel of Miyama. With its egg-shaped base and cloud-shaped handle, "Cloud" exudes a fun coffee time with a free and relaxed mind. A cup holder for espresso machine is included in each package. This product will be released simultaneously in Singapore and Japan. The method of slip casting is used for the production of "Cloud". Slip casting is a type of ceramic molding technique which uses plaster molds instead of using the potter's wheel. By pouring slip into plaster molds and drying it until its hardened, this technique makes it possible to create thin and strong ceramic products. "Risen-yaki" is a decoration technique of applying patterns on porcelain using Japanese paper. Copper plates are used as original template for the paper. Japanese paper with patterns is first applied by the craftsperson by hand on the surface of white porcelain one by one and when the paper is removed the patterns will adhere to the porcelain creating beautiful patterns of lines and texture. The technique of "risen-yaki" adds a special touch to porcelain. In 1846, the technique of transfer printing by copper plates was used for the first time in Japan in Inazu-cho, Mizunami of Gifu prefecture where Miyama is located. To date, only a few potteries in this area are still using this technique to the present day. Casey Chen (b.1971) is a Singaporean designer/ artist who excels in a figurative approach to his work. He had gained international recognition by winning a design award in New York City. "Cloud" is his first collaboration with Japanese craft manufacturers. Miyama Co., Ltd. was established in 1977 in Mizunami area in Gifu prefecture where manufacturers of ceramic tableware for export are concentrated among production areas of Mino ware. The advanced technique that Miyama has developed over decades has enabled them to realized the complex form of "Cloud". The blue color used for the saucer of Cloud reflects a sense of Peranakan culture which is a part of Singapore's multi-cultural heritage. Peranakan design has influences of Chinese, Malay and European sensibilities and the uniqueness of Peranakan style can still be observed in the pastel colored buildings, textile and embroidery around Singapore. This product will be released on the same day both in Singapore and in Japan. Inspired by the 50th year anniversary of the diplomatic relations between the two countries, it will be made available for retail in places like lifestyle shops or museum shops and can be ordered at our website (http://cloudespresso.com/) exclusively for customers in Japan. We can offer image sources of the designer and manufacturer to the media or stores for their sales promotion. HULS is a creative company based in Tokyo and Singapore. Through global website productions, products planning or providing resources and information, we support and help in the overseas expansion of Japanese businesses. We are always on the look out for potential Japanese products/materials manufacturers/ industries for overseas expansion. We are focused on the development of the creative industries in Asia while being centered around Singapore. Under the concept of "Roots & Touch", we aim to establish a platform that will bridge across international borders strategically by connecting different cultures and businesses together. KOGEI STANDARD is a bilingual website for expanding Japanese crafts to overseas by introducing company, people and beautiful crafts. - For more information, please contact:


News Article | January 12, 2016
Site: www.scientificamerican.com

You can find lockers everywhere in Japan, so I was naturally expecting to find some at Tokyo's Hiroo Station. It was something of a shock when there weren't any. Not only did their non-existence mean I had to lug my suitcase through the rain, but it seemed an inauspicious omen given that I was en route to a meeting about a hundred-million-dollar attempt to detect something that physicists are sure must exist, but haven't found. One hundred years ago, Albert Einstein predicted that the fabric of space should be rippling with waves. His image of the universe is one of a taut rubber sheet on which massive objects create curved indentations. Gravity is the result these curves, forcing lighter objects to move towards the more deeply embedded heavier ones. As objects move, the sheet flexes to reflect their new positions, creating oscillations in spacetime that travel outwards as a gravitational wave. These waves are a probe to the Universe’s most secret events, from the interiors of collapsing stars to black holes…or they would be if only we could detect them. Among of the strongest gravitational wave sources imaginable is the merger of two black holes. Yet even this rare event would distort spacetime only enough to briefly change the distance between the Earth and the sun by the width of a hydrogen atom. To put it mildly, this makes detection a serious challenge. One of the best hopes for achieving the necessary sensitivity is light. In a technique known as interferometry, a laser beam is split to send two light waves down perpendicular tunnels several kilometers in length. The waves reflect off mirrors to return to the same position and recombine. The intensity of the newly recombined beam depends on the alignment (or phase) of the peaks and troughs in the two waves. This makes it incredibly sensitive to how far each wave has traveled before it recombines. For gravitational wave detection, the tunnel lengths ensure that the peaks from one wave meet the troughs from the second wave. This is known as destructive interference and it cancels the light of the recombined wave entirely. But when a gravitational wave passes, it distorts the lengths of the tunnels, changing how far each wave travels. The peaks and troughs of the two light waves then no longer perfectly align; the beams no longer cancel each other, and a signal is produced. This technique is behind the US-based detector, LIGO. During 3.5 years of operation between 2005 - 2010, LIGO saw no gravitational waves. However, its sensitivity was only enough to detect the very strongest sources and these are rare occurrences in our Galaxy. Now one century after Einstein’s prediction, LIGO has gotten an upgrade (it's now called Advanced LIGO) and Japan is opening a new detector. It is time for business. Buried 200 m below the Ikenoyama mountain in the Gifu prefecture of Japan, the Kamioka Gravitational Wave Detector (KAGRA) is about to turn on its lasers for the first time. Led by the 2015 Physics Nobel Laurette, Takaaki Kajita, KAGRA’s sensitivity should allow it to detect gravitational waves up to 700 million light years away, ten times further than the previous generation of detectors. Its main target will be binary neutron stars; incredibly dense stellar corpses that emit energy in the form of gravitational waves as their orbits around each other decay. Its enormous range means it's not restricted to events only in our own galaxy, so KAGRA should detect multiple gravitational wave events per year. KAGRA’s underground location minimizes seismic noise from the ever-rumbling Japan. The surrounding gneiss rock is famous for its hardness, causing the mountain to act as a single entity that is resistant to shakes. KAGRA’s second new feature is cryogenic cooling, bringing the system down to an frosty -253° Celsius (20 K), to stifle any thermal vibrations. Such a large-scale interferometer is a new venture for Japan and the next two years will be devoted to testing before full operation begins in 2018. While a positive detection would be an incredible achievement, a single detector cannot determine where the gravitational wave originated. For this, KAGRA will join with Advanced LIGO and two other detectors in Europe to form a global network. But what if KAGRA opens its eyes to nothing? Like the train station lockers, could gravitational waves be expected but non-existent? This could be the most exciting result, since it would imply new physics. While it is unlikely gravitational waves do not exist at all, their strength, frequency or waveform might differ from predictions. Alternatively, the events that produce strong gravitational waves could be less energetic than we suspect. One final note: A rumor is circulating on Twitter that Advanced LIGO has seen some sort of signal. The LIGO team hasn't confirmed it, and even if there really is a signal, it could be an artificial one inserted into the detectors to make sure the data analysis pipeline is working properly. Either way, we're entering a new era in the search for gravitational waves, which is going to give us a new view on the universe.

Loading Gifu collaborators
Loading Gifu collaborators