JP4199009B2 - 皮膚用の食事 - Google Patents
皮膚用の食事 Download PDFInfo
- Publication number
- JP4199009B2 JP4199009B2 JP2002592742A JP2002592742A JP4199009B2 JP 4199009 B2 JP4199009 B2 JP 4199009B2 JP 2002592742 A JP2002592742 A JP 2002592742A JP 2002592742 A JP2002592742 A JP 2002592742A JP 4199009 B2 JP4199009 B2 JP 4199009B2
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- Prior art keywords
- food
- skin
- aloe
- vitamin
- kcal
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
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Images
Classifications
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- 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
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- A—HUMAN NECESSITIES
- 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
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Description
皮膚用の食事は、約310から350kcal/100gを提供する。皮膚用の食事は、約10%の水を含む乾性食である。皮膚用の食事は、以下の材料を含む:
米 〜58%
魚粉 〜30%
繊維 〜5%
ビタミンおよびミネラル 〜6%
(ビタミンCを含む 〜1.25%)
コーン油 〜4%
タウリン 〜0.5%
クルクミン 〜0.5%
アロエ 〜0.06%
乾燥原料を量り、混合し、さらに挽く。次に、乾燥混合物をふるいにかけた後、押出して混合食を形成する。混合食を予備調整器(pre-conditioner)に搬送して、そこで特定の割合で蒸気、水、および油と混合する。個々の粒子全体に亘り均一に水分および温度が運ばれるのに十分な滞留時間、予備調整器内に置いた。次に、予備調整された混合物を、調理および成形のために押出成形機に移す。ダイ圧力は200−1000psigであり、ダイ温度は約90−165℃である。空圧によって乾燥機に成形キブルを搬送する。乾燥温度を130−145℃に設定し、乾燥時間は約17分である。乾燥機を出る際の製品の水分含量は、12%未満である。コーティングの前に乾燥キブルのサイズを揃え(またはふるいにかけ)て、塊および微粉を減らす。キブルをコーティングシステムに供給し、そこで、一定の適用速度で、キブルの表面全体に均一にコーティングを施す。コーティング材料として、ダイジェスト(dogest)およびアロエ抽出粉末と油の混合物が挙げられる。アロエ粉末/油混合物は、固定量の油に、正確な量のアロエ抽出粉末分散させることによって調製する。その混合物をよく混合する必要があり、すなわち、適用する前に、全てのアロエ粉末を油中に均一に分散させる。キブルをアロエ粉末/油混合物で最初にコーティングし、その後室温でダイジェストによってコーティングする。次に、コーティングしたキブルを、コーティング後用の冷却器に移し、そこで、包装前に、コーティングした製品を、その最終水分含量(<12%)、水活性(37℃で<0.7)、および温度(<35℃)に調整する。そこでの滞留時間は約15分である。
本発明の食品を与えた効果を、対照食と比較するため、給餌試験をイヌのパネルで行った。対照食は、本発明の食事からビタミンC、クルクミン、タウリン、およびアロエを除いたものと同じ材料を含んでいた。本発明の食品を与えられたイヌは、対照食を与えられたものと比較して、利点を示した。さらに、本発明の食品を与えて、全く有害な副作用は観察されなかった。
胃腸の健康状態が良好であることが分かっている8匹のネコを用いて試験を行った。試験の開始6ヶ月前に、ネコから寄生虫を取り除き、さらにワクチン接種した。
14日間 対照食
28日間 クルクミン補足食
14日間 対照食
対照食およびクルクミン補足食は、いずれも405kcal/100gであった。対照食は、約4%の水分を含む乾性食であった。対照食は、以下の材料を含んでいた:
鶏肉 〜37%
牛脂 〜10%
米 〜20%
コーンミールおよびグルテン 〜26%
ヒマワリ油 〜3%
醸造用酵母およびビタミン 〜3%
クルクミン補足食は、0.5質量%のクルクミンを含んでいた。クルクミンを乾性対照製品の外面にスプレーした。
血液試料を採取し、血清を調製した。血清を分けて、必要となるまで−20℃で凍結した。支持媒体(アガロースゲル)中で生じる、不透明な沈降環として可視化される抗原−抗体反応に基づき特定タンパク質を定量する方法である、放射状免疫拡散法(RID、Bethyl)によって免疫グロブリンの濃度を測定した。濃度の対数を、沈降環の直径と関係付けることによって、抗原濃度を特定することができる。簡単に説明すると、10μlの血清試料および標準試料(IgGに関して5μl)を、ネコのIgG、IgA、およびIgMに特異的なRIDプレートの中央に移した。そのプレートを3日間インキュベートし、沈降環の直径をRIDリーダーで測定した。標準試料の濃度の対数を、沈降環の直径に対してプロットすることによって標準曲線を作成し、得られた回帰方程式を用いて試験試料の濃度を計算した。
グリース反応によって、血清NOを特定した。グリース反応テストをPromegaから購入し、プロトコルシートに記載されているように実施した。簡単に説明すると、血清試料50μlとNO標準(FCSで希釈)とを、96穴プレートのウェルに2重で添加した。スルファニルアミド溶液50μlを各ウェルに分注し、暗がりにおいて室温で5−10分間プレートをインキュベートした。インキュベーション後、NED溶液50μlを各ウェルに添加し、再度暗がりにおいて室温で5−10分間プレートをインキュベートした。520−550nmに設定したマイクロプレートリーダーを用いて直ぐに各ウェルの光学密度を特定した。
イヌでの試験は、ビタミンC補足食(40mg/400kcalの濃度でビタミンCを含む)を与えると、動物の抗酸化体質を向上させ、健康上の利点の増強に貢献することを示した。
方法
20匹の小型犬(すなわち、ミニチュアシュナウザー、ケアーンテリア、コッカースパニエル、プードル、またはウェストハイランドホワイトテリア)を用いて実施した。イヌの年齢/性を一致させ、試験の間、それぞれの囲いの中で個別に食事を与えた。自然の日光周期で、室温を22℃に維持した。
この測定法は、分散法を用いて評価したバリア機能の改善に対する、ビタミンC、タウリン、アロエ、およびクルクミンを含む食品の効果について立証する。放射性標識した水がin vitro皮膚バリア(イヌのケラチノサイト皮膚細胞から構成される)を越えて分散する速度を、本食品の存在下または不在下で培養した細胞において比較した。
Costar Snapwellプレート(ASL Cat No. 402/0369/08)を、外側ウェルに2.6mlのGreens培地、内側のウェルに400μlのGreens培地を含むように設定し、内側ウェルにイヌのケラチノサイトを1x105播種した。そのプレートを37℃、5%CO2でインキュベートした。内側ウェル中のGreens培地は次の日に交換して死細胞を除いた。そのプレートをさらに2日間培養した。試験濃度の食品(10μl/ml)を含むようにGreens培地を調製した。対照培地は、DMSO(10μl/ml)を含むように調製した。4日目に、内側および外側のウェルから培地を除いて、900μlの試験/対照培地を外側ウェルに入れた。食品の濃度当り2個のスナップウェルを用い、プレートごとに1つの対照を用いた。このような低レベルの培地は、ケラチノサイトが、空気−液体境界面に存在することを確実にした。2−3日ごとに培地を交換しながら、プレートをさらに7日間培養した。11日目に、分散法を実施するためにスナップウェルの用意ができた。各スナップウェルの内側ウェルを取り除いて、個々の分散チャンバーに置いた。ダルベッコ変法イーグル培地(DMEM)をチャンバーの両側に添加(6mlずつ)し、各チャンバーを分散装置に置いた。分散装置はチャンバーを37℃に平衡化させ、したがって培地の動きを確実にした。100μlの放射性標識(3H)した水を各チャンバーの左側に添加し、3分間隔で90分間、右側から50μlの試料を採取した。各試料を採取した後、DMEM(50μl)を右側に入れた。4mlのシンチレーション液を含有するシンチレーションバイアルに試料を入れて、各試料中の放射性標識の量をシンチレーションカウンターでカウントした。
イヌのケラチノサイト皮膚細胞が、層状脂質であるセラミドを合成する能力について、食品の存在下および不在下で培養した細胞において比較した。セラミドの合成の増加は、皮膚の層状構造を改善することによって、皮膚のバリア機能を改善する。14C−セリン取込みレベルを測定することによって、セラミド合成を直接評価した。
イヌケラチノサイトを、5x104/ウェルの細胞密度で、MCDB153培地(以下の内容を参照)中、コラーゲンコート24穴プレート(Sigma, Cat No. Z38,049-0)に播種した。そのプレートを、37℃、5%CO2でインキュベートした。次の日、各ウェルにつき培地を交換し、死細胞を除去した。インキュベーション4日目に、再度培地を交換した(500μlのBPEを含まないMCDB153)。次に、様々な試験濃度で食品(10μl/ml)を添加し、対照ウェルにはDMSO(10μl/ml)を添加した(試験/対照当り、6ウェルを用いた)。食品添加の後、さらに5日間インキュベートし、この間に1回培地および食品を交換した。5日目に、培地および食品を再度交換し、5μlの14C−セリンを各ウェルに添加した。そのプレートをインキュベートし、12日目にトリプシンを用いて細胞を回収した。各ウェルから別々に回収し、ペレットを−20℃で保存した。Bligh-Dyer溶剤を用いて、各細胞ペレット中への14Cセリンの取込みを測定した。細胞ペレットを解凍し、300μlのBligh-Dyer 溶剤を各ペレットに添加した(クロロホルム10ml、メタノール5ml、脱イオン水1ml、および0.88% KCl1ml)。電動式ペレット乳棒を用いて、その溶液を20秒間混合した。次に、試料を1300rpmで3分間回転させて、層の分離を促進させた。次に、各試料の底層(放射性標識脂質を含む)を注意深く取り除き、4mlのシンチレーション液を含むシンチレーションバイアルに加えた。各試料中の14C−セリンの量をシンチレーションカウンターで測定した。
MCDB153(Sigma#M7403)
インシュリン(5mg/L)、ヒドロコルチゾン(180mg/L)、2−アミノエタノール(6.1mg/L)、O−ホスホリルエタノールアミン(14.1mg/L)、上皮成長因子(100ng/L)、およびウシ下垂体抽出物(0.4%v/v)
インシュリン:Sigma#I6634
ハイドロコルチゾン:Sigma#H0396
2−アミノエタノール:Sigma#E0135
O−ホスホリルエタノールアミン:Sigma#P0503
上皮成長因子:Sigma#E4127
ウシ下垂体抽出物:Sigma#P1476/Invitrogen#13028-014
結論
皮膚におけるセラミド合成のレベルは、食品の存在下で培養した細胞において上昇した。このデータは、食品の存在下での細胞のインキュベーションが、層状脂質合成のレベルを高めることによってバリア機能を改善し、それによって、高められたバリア機能を有する皮膚を作るのを助けることができることを示唆する。言い換えると、このことは、それぞれ感染またはアレルギーにつながる、病原菌またはアレルゲンの皮膚を介した還流を防止するであろう。
本測定は、皮膚回復測定法を用いて評価した、in vitroでの創傷における皮膚回復に対する食品(ビタミンC、タウリン、アロエ、クルクミン)の効果を特定する。イヌの皮膚繊維芽細胞が傷ついた後遊走する能力を、食品の存在下および不在下で培養した細胞において比較した。
本実験の1日目に、組織培養ディッシュ(蓋および通気孔を有する、滅菌、35mmx10mm、2x2mm格子、174926Nalge Nunc international)を用いて、傷線を表すように中心線のほぼ下に各ディッシュの裏面に線を引いた。2枚のディッシュを対照用に割り当て、また2枚を試験用に割り当てた。次に、イヌの皮膚繊維芽細胞(2mlの繊維芽細胞用培地中、約4x105/ディッシュ)をディッシュに播種し、37℃で一晩インキュベートした。
傷をつけることによって作成した裸スペースに遊走した細胞数は、食品の存在下で培養した細胞において増加した。このデータは、食品の存在下での細胞の培養が、おそらく細胞の遊走および/または細胞の増殖を促進させることによって、皮膚の回復を促進させることを示す。したがって、バリア機能が改善され、かつケガまたは病気後により高い回復能力を有する皮膚を作るのを助ける。
1)大腸菌、Staph I、SC01−SC08を、BHI寒天プレート上、38℃で一晩インキュベートする;
2)次の日、菌株ごとに、細菌の1コロニーを10mlのBHIブロスに入れて、38℃で静的にインキュベートした;
3)以下の材料、下記に概説するビタミンC、タウリン、アロエ、およびクルクミン、から構成される食品を、1mg食品/1ml BHIブロスで、50mlのBHIブロス中に調製する。次に、激しくボルテックスで攪拌し、その後38℃でインキュベートした;
4)3時間後、10倍連続希釈を行い、増殖指標プレート(growth indicator plate)(以下を参照)を作成する;
5)各菌株に関して各細菌ブロスを2つに分け、新しいチューブに5mlを入れ、残りの5mlを元のチューブに残す;
6)4段目のチューブの一方に、5mlの新鮮なBHIを添加する(対照);
7)4段目のもう一方のチューブに、5mlの食品(1mg/ml)を添加する(試験);
8)各菌株に関して両方のチューブ(対照および試験)が、最終容積10mlを含む。次に、チューブを38℃でインキュベートする。
10)一晩インキュベートした後、再度指標プレートを作成する。
1)ビタミンC:30mg/400kcal
2)タウリン:500mg/400kcal
3)アロエ:70mg/400kcal
4)クルクミン:500mg/400kcal
試験する食品の濃度
0.1mg/ml−20mg/mlの濃度で食品を試験した。それら濃度は、平均サイズのイヌに対して1日当り300gの食事を与えた後の血中において利用できる食品の量に相当する。
Staph I:増殖を減少させる僅かな効果が6時間後に観察されたが、24時間後には観察されなかった。24時間後には、増殖の僅かな増加が観察される。これは、本発明の食品がStaph Iに対して静菌性を有することを示唆する。
アトピーのようなアレルギー性皮膚症状において、プロスタグランジンE2およびロイコトリエンB4のような炎症性メディエーター(pro-inflammatory mediator)の循環レベルが上昇する傾向がある。このことは、アトピー部位において炎症を増強させることにつながり、回復プロセスに混乱を生じさせる。この方法は、イヌ皮膚繊維芽細胞によって産生される上清PGE2の濃度に対する食品補足の効果を調べる。
本実験に関して、前述した回復測定からの細胞上清を用いた。PGE2高感度測定キット用のR+Dシステム(cat No. DE2100)のハンドブックに記載されている方法を用いて上清を分析した。
ダックスフント(6才、メス)は、4年間皮膚症状を患っている。過去に、その症状を緩和するために、ステロイド、抗ヒスタミン剤、脂肪酸、抗生物質、向精神薬、除外食、シャンプー、および耳の処置を含む多くの処置を行った。魚、肉、鶏、および菜食を含む様々な食事も用いた。
Claims (26)
- ビタミンC、タウリン、クルクミン、およびアロエを含む食品。
- ビタミンAをさらに含むことを特徴とする請求項1記載の食品。
- 亜鉛をさらに含むことを特徴とする請求項1または2記載の食品。
- 1以上の脂肪酸をさらに含むことを特徴とする請求項1から3いずれか1項記載の食品。
- 前記脂肪酸が、エイコサペンタエン酸、ドコサヘキサエン酸、α−リノレン酸、およびγ−リノレン酸より成る群から選択されることを特徴とする請求項4記載の食品。
- 400kcal当り、20mgから500mgのビタミンCを含むことを特徴とする請求項1から5いずれか1項記載の食品。
- 400kcal当り、100mgから1000mgのタウリンを含むことを特徴とする請求項1から6いずれか1項記載の食品。
- 前記アロエが、100%活性固形物として含まれることを特徴とする請求項1から7いずれか1項記載の食品。
- 400kcal当り、1mgから1000mgのアロエを含むことを特徴とする請求項1から8いずれか1項記載の食品。
- 400kcal当り、100mgから1000mgのクルクミンを含むことを特徴とする請求項1から9いずれか1項記載の食品。
- 400kcal当り、1000IUから10,000IUのビタミンAを含むことを特徴とする請求項2から10いずれか1項記載の食品。
- 400kcal当り、5mgから50mgの亜鉛を含むことを特徴とする請求項3から11いずれか1項記載の食品。
- 400kcal当り、10mgから1000mgの1以上の脂肪酸を含むことを特徴とする請求項4から12いずれか1項記載の食品。
- 医薬において用いられることを特徴とする請求項1から13いずれか1項記載の食品。
- 皮膚障害を制御するのに用いられることを特徴とする請求項14記載の食品。
- 炎症性またはアレルギー性皮膚反応を制御するのに用いられることを特徴とする請求項14または15記載の食品。
- 前記皮膚反応が、アトピー、食物アレルギー、接触アレルギー、皮膚炎、ノミアレルギー、痒み、脱毛、または炎症の1以上であることを特徴とする請求項16記載の食品。
- 炎症性またはアレルギー性皮膚反応に関連する細菌感染を制御するために用いられることを特徴とする請求項1から17いずれか1項記載の食品。
- 皮膚障害の制御のための組成物の製造における、ビタミンC,タウリン、クルクミン、およびアロエの使用。
- 前記皮膚障害が、炎症性またはアレルギー性皮膚反応であることを特徴とする請求項19記載の使用。
- 前記皮膚反応が、アトピー、食物アレルギー、接触アレルギー、皮膚炎、ノミアレルギー、痒み、脱毛、または炎症の1以上であることを特徴とする請求項20記載の使用。
- 炎症性またはアレルギー性皮膚反応に関連する細菌感染を制御するための使用であることを特徴とする請求項19から21いずれか1項記載の使用。
- 動物(ヒトを除く)において皮膚障害を制御する方法であって、請求項1から13いずれか1項記載の食品を前記動物に与える工程を含む方法。
- 前記皮膚障害が、炎症性またはアレルギー性皮膚反応であることを特徴とする請求項23記載の方法。
- 前記皮膚反応が、アトピー、食物アレルギー、接触アレルギー、皮膚炎、ノミアレルギー、痒み、脱毛、または炎症の1以上であることを特徴とする請求項24記載の方法。
- 炎症性またはアレルギー性皮膚反応に関連する細菌感染を制御するための方法であることを特徴とする請求項23から25いずれか1項記載の方法。
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GBGB0113348.7A GB0113348D0 (en) | 2001-06-01 | 2001-06-01 | Skin diet |
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2001
- 2001-06-01 GB GBGB0113348.7A patent/GB0113348D0/en not_active Ceased
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2002
- 2002-03-27 AU AU29174/02A patent/AU785003B2/en not_active Expired
- 2002-05-31 JP JP2002592742A patent/JP4199009B2/ja not_active Expired - Lifetime
- 2002-05-31 DK DK02738346T patent/DK1392131T3/da active
- 2002-05-31 EP EP02738346A patent/EP1392131B1/en not_active Expired - Lifetime
- 2002-05-31 DE DE60228235T patent/DE60228235D1/de not_active Expired - Lifetime
- 2002-05-31 GB GB0212767A patent/GB2378133B/en not_active Expired - Lifetime
- 2002-05-31 US US10/479,065 patent/US8647681B2/en active Active
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WO2002096221A2 (en) | 2002-12-05 |
DK1392131T3 (da) | 2008-11-17 |
CA2449146C (en) | 2012-04-24 |
ES2312583T3 (es) | 2009-03-01 |
EP1392131A2 (en) | 2004-03-03 |
AU2917402A (en) | 2002-12-05 |
CA2449146A1 (en) | 2002-12-05 |
DE60228235D1 (de) | 2008-09-25 |
US8647681B2 (en) | 2014-02-11 |
JP2004520848A (ja) | 2004-07-15 |
GB0113348D0 (en) | 2001-07-25 |
GB2378133B (en) | 2005-01-26 |
GB2378133A (en) | 2003-02-05 |
ATE404075T1 (de) | 2008-08-15 |
US20040241286A1 (en) | 2004-12-02 |
EP1392131B1 (en) | 2008-08-13 |
AU785003B2 (en) | 2006-08-24 |
WO2002096221A3 (en) | 2003-01-16 |
GB0212767D0 (en) | 2002-07-10 |
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