JP6842072B2 - エガセラ属細菌の菌数抑制剤 - Google Patents
エガセラ属細菌の菌数抑制剤 Download PDFInfo
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- JP6842072B2 JP6842072B2 JP2019041539A JP2019041539A JP6842072B2 JP 6842072 B2 JP6842072 B2 JP 6842072B2 JP 2019041539 A JP2019041539 A JP 2019041539A JP 2019041539 A JP2019041539 A JP 2019041539A JP 6842072 B2 JP6842072 B2 JP 6842072B2
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- kestose
- bacteria
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- 238000001356 surgical procedure Methods 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 150000004044 tetrasaccharides Chemical class 0.000 description 1
- 231100000820 toxicity test Toxicity 0.000 description 1
- LOIYMIARKYCTBW-OWOJBTEDSA-N trans-urocanic acid Chemical compound OC(=O)\C=C\C1=CNC=N1 LOIYMIARKYCTBW-OWOJBTEDSA-N 0.000 description 1
- LOIYMIARKYCTBW-UHFFFAOYSA-N trans-urocanic acid Natural products OC(=O)C=CC1=CNC=N1 LOIYMIARKYCTBW-UHFFFAOYSA-N 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- A61K31/00—Medicinal preparations containing organic active ingredients
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- A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
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- A61P25/00—Drugs for disorders of the nervous system
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- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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Description
(1)糞便含有微生物の培養
既報〈E Olano−Martinら、Br J Nutr、第83巻、第247−255頁、2000年〉に従って下記の組成の嫌気緩衝液および基本培地を調製した。また、1−ケストース水溶液(終濃度0.5%(w/v)を調製した。嫌気緩衝液、基本培地、1−ケストース水溶液、滅菌水および硝子瓶は窒素ガスで浄化した後、オートクレーブ滅菌を行った。
《嫌気緩衝液の組成/1Lあたり》(最終pH:pH7.0)
ペプトン1g、塩化ナトリウム8.5g、システイン塩酸塩0.5g。
《基本培地の組成/1Lあたり》(最終pH:pH7.0)
ペプトン2g、酵母抽出物2g、塩化ナトリウム0.1g、リン酸水素二カリウム0.04g、リン酸二水素カリウム0.04g、硫酸マグネシウム七水和物0.01g、塩化カルシウム二水和物0.01g、炭酸水素ナトリウム2g、システイン塩酸塩0.5g、胆汁酸塩0.5g、ヘミン0.005g、Tween 80 2mL、フィロキノン10μL、レザズリンナトリウム。
〈Shunsuke Takahashiら、PLosONE、第9巻、第8号、e105592、2014年8月:参考文献1〉に記載の方法に従って、本実施例1(1)の培養前検体および培養後検体から総DNAを抽出した。具体的には、まず、4Mのグアニジンチオシアネート、100mMのトリスHCl(pH9.0)および40mMのEDTAを含む水溶液に、氷上で融解した培養前検体または培養後検体100mgを懸濁し、FastPrep FP100A(MP Biomedicals)を用いてジルコニアビーズで粉砕して懸濁液を得た。Magtration System 12GC(Precision System Science)およびGC series MagDEA DNA 200(Precision System Science)を用いて、この懸濁液からDNAを抽出した。DNAの濃度を分光測光器ND-1000(NanDrop Technologies)を用いて測定し、10ng/μLとなるように調製して、これを糞便由来総DNAとした。
上記参考文献1に記載の方法に従い、本実施例1(1)の培養前後の糞便に含まれる細菌の種類と存在比率の網羅的解析を行った。すなわち、まず、本実施例1(2)の糞便来総DNAを鋳型として、下記配列番号3および4のユニバーサルプライマーを用いてDual−index法(Hisada Takayoshiら、Arch Microbiol、197巻、第7号、第919‐34頁、2015年6月)によりポリメラーゼ連鎖反応(PCR)を行い、細菌由来の16S rDNAのV3−V4領域を増幅した。
フォワードプライマー(341f):CCTACGGGAGGCAGCAG(配列番号3)
リバースプライマー(R806):GGACTACHVGGGTWTCTAAT(配列番号4)
健常者(23〜62歳、男性、体重60.0〜98.6kg)38名を、ケストース群20名、プラセボ群18名の2つの群に分けた。ケストース群には1−ケストースを、プラセボ群にはマルトースを、試験期間中、毎日経口摂取させた。なお、試験期間中、被検者には、1−ケストースまたはマルトースのいずれを摂取しているのか不明な状態とした。試験期間は12週間、1−ケストースまたはマルトースの1日の摂取量は10g(約0.17〜0.10g/kg体重/日)とした。試験期間の開始時および終了時に糞便を採取し、−80℃にて凍結保存した。
Claims (3)
- 1−ケストースを有効成分とする、エガセラ属細菌の菌数抑制剤。
- エガセラ属細菌による菌血症、副鼻腔炎、化膿性筋炎、皮膚膿瘍、脊椎椎間板炎および肝膿瘍から選択される疾患の予防または治療に用いられることを特徴とする、請求項1に記載の菌数抑制剤。
- 1−ケストースを有効成分とする、エガセラ属細菌の菌数抑制用食品組成物。
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