この記事は伊賀/FT/2017/007 zlin 大学のプロジェクトのサポートをしました。

ボランティアは、従業員や私たちの大学の学生の間で採用されました。選択する方法によると行った「医の国際倫理指針関与人間被験者。懇談会 (2002) ジュネーブ医科学国際組織。」KH はヘアケア製品(シャンプー、コンディショナー等)で使用される一般的な化粧品の成分とそれ故に制度上の審査委員会からの承認は必要ではありません。
1. KH にプロセス鶏羽
<ol><li>養鶏場の鶏の羽を収集します。</li>
	<li>十分な過剰の新鮮な (コールド) を実行している水と鶏の羽根から血液の残骸や不溶性不純物を洗い流します。平らなプレートに羽を置き、50 ° C で一晩乾燥 注: プロトコルはここで一時停止することができます。</li>
	<li>また最終的な細かさ 1.0 mm. に 50 g 乾燥羽 (ミディアム ハード、ソフトのサンプル材料、繊維材料に適した) による切削を挽く、研磨羽の最終的な細かさが高い、しかし、3.0 mm にすることができます。 注: プロトコルはここで一時停止することができます。</li>
	<li>
羽を脱脂します。 注: 家禽の羽を脱脂の最も効果的かつ経済法は、脂肪分解酵素を使用しています。<ol>
<li>温度制御ステンレス鋼 27 L ボイラー容器、水 40 ± 2 ° C の重量比 1: 75 まで予熱で羽をミックスします。脂肪分解酵素を追加 (活動 100 KLU/g) ゆっくりと 1.5-2.0% (で重量を量った乾燥羽に関連) の量の 5 分のオーバーヘッド攪拌で内容をかき混ぜます。</li>
		<li>9.0 ± 0.2 1% 水酸化ナトリウムまたは 1% H3PO4ソリューションを追加することにより、脂肪分解酵素の活性の最大値に対応する値に混合物の pH を調整します。オーバーヘッドの攪拌器と 5 分の混合物をかき混ぜるし、チェックして再実験室ベンチ mV/pH メーターを使用して ph を調整します。</li>
		<li>軽く 40 ± 24 h のオーバーヘッド攪拌機で混合物をかき混ぜる 0.5 の ° c.また、40 ± 0.5 ° C で混合物を孵化、孵化の最初の 6 時間の間に 1 時間間隔で内容をかき混ぜます。</li>
		<li>(100 μ m サイズ) 目の細かいふるいを通して混合物をフィルターし、新鮮な (コールド) を実行している水の流れと脱脂の羽を洗ってください。一晩乾燥室で 50 ° C で平板に羽を乾かしてください。 注: プロトコルはここで一時停止することができます。</li>
	</ol>
</li>
	<li>鶏羽毛加水分解の最初の段階を実行します。羽 0.3 %koh 水溶液を混ぜて、重量比 1:50 に、60 ± 0.5 ° C 24 時間オーバーヘッド攪拌で軽くかき混ぜます。混合物の pH は、インキュベーションの終わりに約 11.0 培養開始時約 12.5 から減る。加水分解の最初の段階を終えた後に %1 を追加することによって 10% H3PO4 (この場合は、9.0 ± 0.2 のレベル) にタンパク質分解酵素の活性の最大値に対応するレベルの混合物の pH を調整水酸化ナトリウム。</li>
	<li>鶏羽毛加水分解の第 2 段階を実行します。5.0% (秤量の加水分解の最初の段階の初めに羽の量の乾物に関連) の投与量でのタンパク質分解酵素の混合物に追加します。優しく 60 ± 0.5 ° C、8 h でオーバーヘッド攪拌でかき混ぜる沸点 (100 ° C) (同じステンレス 27 L ボイラー コンテナー) の混合物を加熱し、酵素を不活化する 10 分煮る。</li>
	<li>わずかな真空圧力; でブフナーの低密度のフィルター ペーパーを通してフィルタ リングによって不溶残から KH (1.6 の手順で準備) のソリューションを分離します。また、遠心分離機を使用します。 注: プロトコルを一時停止できるここ数日間 5 ± kh 社のソリューションが格納されている場合 1 ° C</li>
	<li>プラスチック製のバケツ (直径 x 26 cm 高さ 26 センチ) で小さなペプチドと塩を削除する 12 K MWCO 膜を用いた KH を dialyze します。透析チューブに KH 溶液 400 mL を注ぎ、室温; 80 h の蒸留水 4 L に対して dialyze18、36、および 60 h 後蒸留水を変更します。</li>
	<li>1,000 cm2プレートの面積、真空乾燥 40 ± 0.5 ° C で薄膜に一晩、微粉末に粉砕、乾燥器で密閉容器に保管する 500 mL の比 (例えばシリコーン) 付着防止プレートに KH の透析解決にキャストします。 注: プロトコルを一時停止できるここ数ヶ月間瀬粉は乾燥した場所に格納されている場合。</li>
</ol>2. kh さん化粧品製剤を準備します。
注: テスト用 OB だった商業親水性 O/W クリーム ベース、アクア、パラフィン、パラフィン液相、セテアリル アルコール、ラウレス 4、水酸化ナトリウム、カルボマー、メチルパラベン、プロピルパラベン ・で構成します。
<ol><li>2、4、および 6% を含む製剤を準備 (軟膏の基本重量) に従い KH。ポリエチレン容器 (直径 x 10 cm 高さ 7 センチ) に瀬粉の量の重量を量るし、定式化の総重量を保証する金額で OB に等しい 50 g 追加表 1のレシピを参照してください。</li>
</ol><table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td>香粧品配合</td>
			<td colspan="3">軟膏ベース [g] の重量</td>
			<td colspan="3">ケラチン加水分解物 【 g の重量</td>
			<td colspan="2">総重量 [g]</td>
		</tr>
<tr>
<td>軟膏ベース</td>
			<td colspan="3">50</td>
			<td colspan="3">0</td>
			<td colspan="2" rowspan="4">50</td>
		</tr>
<tr>
<td>軟膏ベース + 2% KH</td>
			<td colspan="3">49</td>
			<td colspan="3">1</td>
		</tr>
<tr>
<td>軟膏ベース + 4% KH</td>
			<td colspan="3">48</td>
			<td colspan="3">2</td>
		</tr>
<tr>
<td>軟膏ベース + 6% KH</td>
			<td colspan="3">47</td>
			<td colspan="3">3</td>
		</tr>
</tbody></table>表 1: 重量の量化粧品処方を準備する軟膏ベースとケラチン加水分解物。
<ol start="2"><li>134.16 x g で 10 分間の 3 刃研究室ミキサーで混合物を均質化し、機械的頭上式撹拌機を使用してミックスします。5 ± 1 ° C で準備された製剤を維持し、2 h 使用する前に室温にそれらを暖めます。 注: kh さん OB の混合物を均質化は、非デジタルの攪拌機と同様に実行できます。非デジタル攪拌にも (rpm) でおおよその速度と体重計があります。穏やかな攪拌がこのステップの最高を仕事します。 注: プロトコルを一時停止できるここまで 5 ヶ月製剤が 5 ± で格納されている場合 1 ° C</li>
</ol>3. 肌の水分補給を測定、TEWL、pH、KH のプロパティのテストします。
注: は、23 ± 2 ° C でエアコン ルームと 56 ± 3% の相対湿度ですべての測定を実行します。
<ol><li>場所 5 のフィルター ペーパーのストリップ (2 × 4 cm サイズ) 生理のソリューションに (0.90 %nacl) とソリューションで約 1 分間のまま。</li>
	<li>右前腕と左の前腕の内側に 3 つの内側に 2 つのストリップを適用、4 h の絆創膏で固定します。この手順は、皮膚を脱脂してサイトのスキンの個々 の特性を排除することです。4 h 後ストリップを削除し永続的なペンを持つデンマークの領域、図 1を参照してください。</li>
</ol><img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56254/56254fig1.jpg"> 図 1: 左と右の手足の前腕に試験製剤の位置の方法。<a href="//cloudflare.jove.com/files/ftp_upload/56254/56254fig1large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。
<ol start="3"><li>0.1 mL 注射器を使用して脱脂前腕サイトの各スポットでテスト済みの製剤を適用し、マークされた表面全体に 。左の前腕にしない (それはコントロール) 最初のサイトに何かを適用、2 番目のサイトに OB と OB + 2 を適用第 3 %kh。適用 OB + 4 %kh と OB + 6% 右腕に KH。</li>
	<li>各サイトと (1、2、3、4、24、および 48 h) 各間隔で各サンプルを測定し、肌の水分補給、肌 TEWL、TEWL 計プローブと 15 の測定値と皮膚 pH の皮膚 pH 計プローブを読んで 1 皮膚水和計プローブ 5 測定値を取る。テスト期間中にテスト現場や周辺地域にどんな化粧品を適用するボランティアを許可しません。簡単な夜洗浄水を実行してもかまいません。 注: プロトコルはここで一時停止することができます。</li>
	<li>記述統計、表計算ソフトを使用しての基本数値特性による結果の測定値を処理します。5 の水和は、各サンプルの測定を測定値から最低および最高の測定値を無視し、のみ 3 測定のための算術平均と標準偏差を計算します。各サンプルの測定 15 TEWL 測定から最初の 5 を無視してのみ 10 測定のための算術平均と標準偏差を計算します。</li>
</ol>

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K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transdermal+permeability+and+skin+accumulation+of+amino+acids.">Transdermal permeability and skin accumulation of amino acids.</a> Int. J. Pharm. 72 (2), 149-155 (1991).</li><li>Draelos, Z. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Therapeutic+moisturizers.">Therapeutic moisturizers.</a> Dermatol. Clin. 18 (4), 597-607 (2000).</li><li>Courage and Khazaka Electronic GmbH, Technical Charges. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Information and Operating Instructions for the Multi probe Adapter MPA and its Probe. , (2013).</li></ol>

著者が明らかに何もありません。

アルカリ酵素加水分解の利点は、kh 社の今後の用途に合わせて変更できることです。たとえば、製品の軽く茶色の色が障害ではなく、ヘアケア化粧品のアプリケーションで加水分解高温適用できます KH の歩留まりの向上に 。さらに、技術のプロシージャの両方の段階の間に長い処理時間に大きく影響プロセス全体効率-KH の収量が 85% に上昇します。
水和測定結果は、測...

食肉処理場、食品工業および日焼け業界年間固体ケラチン副産物-ウール、羽、毛、ひづめの音、爪、角、等の膨大な量を生成します。最新の統計データによると、ライブの総重量鶏、七面鳥、アヒル、および米国の他の虐殺された家禽は1年あたり 625 億ポンドEU では年間約 287 億ポンドです。羽の家禽の合計重量の 8.5% までことを考えると、単独で米国は毎年廃棄物の羽2の約 53 億ポンドを生成します。
ケラチンは、強くとレンダリングの処理が困難なジスルフィド架橋だから耐薬品性を示すタンパク質です。水溶性の製品を取得するには、架橋結合を切断し、おそらく3ペプチド結合の加水分解を実施する必要があります。二硫化物橋の胸の谷間は、次のパターン4,5によるとチオール アニオンの反応を続行可能性があります。
S、- + -SbSc-↔ – Sb- + -S、S のc-
非常に高い ph で、ジスルフィド結合の加水分解も表示され、パターン6に従い
-SOH + オハイオ州- →-S- -- SS
(PH 約 8) 温和な条件下でも sulfitolysis は、次のパターンに従って起こる。
-SS- + HSO3- →-SH + SSO-3-
ケラチンを分解する最も経済的な方法は微生物の内訳は、加工や高分解効率 (約 90%)7,8の間に穏やかな反応条件によって特徴付けられる。Keratinases は、土壌とケラチン廃棄物9から分離されたいくつかの細菌によって生成されます。微生物 keratinases は堅く、強く架橋ケラチン構造10を加水分解し、準備結果 KH はタンパク質が豊富で水溶性、検出された必須アミノ酸の損失なしで11。
このようなタンパク質は水に溶け、与えられたシステムが透明で、ペプチドの再集計は避けるために、化粧品の準備 (例えば乳液、ローション、ゲル) で蛋白質を組み込むために要件を確保します。疎水性相互作用。したがって、加水分解コラーゲン、エラスチン、ケラチンなどのタンパク質の加水分解物に適用する一般的な勧めします。化粧品エマルジョンに加水分解液を追加するときは、手順を実行して、加水分解は最初水に溶解したことを確認します。いくつかの場合、望ましい蛋白質 (または、加水分解物)、アルコールまたは12の他の有機溶剤に可溶です。
KH は、シャンプー、コンディショナー、ローションと髪の毛と同様、マスカラー、マニキュア、目メイク エージェントに栄養血清で通常紹介されています。通常宣言 KH 効果髪を平滑化、保護膜の形成が含まれてまたは爪の構造、高まり可塑性と製品の整合性を調節し、泡13の形成を助長、処理層の外観,14. それが KH 低減表面張力、化粧品で補充が乳化剤クリームを安定させるために追加量の削減を促進するそれ故にも示されています。KH は肌、目、髪、洗浄剤 (界面活性剤) によって引き起こされる炎症の影響を制限し、洗浄剤 (例えば脱水肌、硬度および減らされた柵機能の組織上の任意の潜在的な副作用を減らす肌のために)。加水の高いバッファリング機能は、化粧品の pH を安定させるために悪用も長さの短いペプチドがある大きい緩衝効果15,16。KHs は、髪と爪化粧品だけでなく、皮膚のケアのための製品で利用されている、標準コンポーネントとして定着した、KH の保湿の効果に関する研究は現代文学で表示されません。
アルカリ酵素技術、KH にケラチン副産物を処理するためには、化粧品添加剤17,18,19,20数の影響上のプロセスでは、アクティブなテスト,21,22. 2 段階アルカリ酵素加水分解微生物プロテアーゼを用いた鶏の羽根の利点が穏やかな反応条件下で高効率を実現、kh 社の品質は非常に強い酸で加水分解とは対照的に高またはアルカリ。最初の段階で羽に部分的にケラチン構造を混乱させるし、うねり; 羽アルカリ環境で高温孵化します。pH を調整した後羽の 2 番目の段階で温和な条件下でのタンパク質分解酵素で加水分解します。透析の瀬では、蛋白質の高いコンテンツを所有しています。
ここで説明したメソッドの目的はアルカリ酵素加水分解 KH に家禽の羽の処理、乳化化粧品に適用される KH の保湿のプロパティの効果をテストします。体内機器の非侵襲的方法、保湿特性を調べた。SC の皮膚水分補給とバリア機能を測定するための最も頻繁な方法には (コンダクタンスや容量) の皮膚の電気的特性の測定が含まれます。赤外マルチ スペクトル想像法 (NIM)、核磁気共鳴分光法、光干渉断層計、または過渡的熱伝達23近く SC 水和反応を調査するための異なる方法があります。SC の TEWL に相関する SC のバリア機能と換気チャンバー法、非換気チャンバー オープン商工会議所法24測定されます。
配合で、モデルのプロパティは、3 種類のプローブのマルチ プローブ アダプター MPA 5 を使用して決定されます。最初の 1 つ、corneometer CM 825、対策皮膚水和皮膚の表面の電気容量の変化を評価することによって測定コンデンサーは corneometric 単位で皮膚表面の静電容量の変化を示しています。Corneometer は、皮膚水和25の相対評価だけを与えます。TEWL の 2 番目のプローブ、tewameter TM 300 が直接使用されるない皮膚から (Fick の拡散の法律に基づいて開いた区域楽器) で蒸発する水の密度勾配を測定するセンサー (温度・湿度) の 2 つのペアで定義されているエリア、時間帯 (g/m2/h) ごと運ばれる水の量を示します。このメソッドは、皮膚バリア機能26のもわずかな中断を検出できます。皮膚の pH は障壁の一つの指標と SC27の抗菌機能です。皮膚マントルの酸性度は、MPA 5 駅に接続された (3) 皮膚 PH 905 プローブで測定しました。この特別に設計されたプローブから成っている平たいガラス電極による皮膚への接触を電圧計に接続されています。システムは、プローブの上部に測定した半固形状の非常に薄い層を取り巻く水素陽イオンの活動のための潜在的な変化を測定します。電圧の変化は pH28として表示されます。
3 つのセクションに分かれて実験を提案する: 化粧品の準備 (2) の (塩と低分子画分を除去する)、透析による 2 段階アルカリ酵素加水分解し精製羽鶏から KH の準備 (1)2、4、および 6% を含む製剤 TEWL、水和を皮膚や肌の pH、KH と KH のプロパティを測定することによってテストする (3)。実施した平均年齢 27.2 歳で 10 人の女性と平均年齢 26.2 歳の 10 人。ボランティアとテスト自体の選択方法を行った被験者29; を利用した生物医学研究の国際的倫理的原則に従いすべての人は、研究の包含前にインフォームド コンセントを与えた。開始、テスト前にボランティアは、それらの健康状態に関するアンケートを完了するように求めていた。ボランティアの前に 24 時間の間に、テスト期間中にテスト サイトと周辺地域にどんな化粧品を適用しないようにすることを約束さらに、彼らはのみ水を実行して簡単な夜洗浄を許されました。

<table>
	<tbody>
		<tr>
			<td>Material or chemicals</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>LIPEX 100T</td>
			<td>Novozymes</td>
			<td>LJP30020</td>
			<td>Lipex - enzyme produced by submerged fermentation of a genetically-modified microorganism, activity 100 KLU/g</td>
		</tr>
		<tr>
			<td>Savinase Ultra 16L</td>
			<td>Novozymes</td>
			<td>PXN40001</td>
			<td>Savinase - enzyme produced by submerged fermentation of a genetically-modified microorganism, activity 16 KNPU-S/g</td>
		</tr>
		<tr>
			<td>Potassium hydroxide, KOH</td>
			<td>Sigma-Aldrich</td>
			<td>302510289</td>
			<td>Potassium hydroxide, KOH, 97,0 %, Mr 56,11</td>
		</tr>
		<tr>
			<td>Phosphoric acid solution, H3PO4</td>
			<td>Sigma-Aldrich</td>
			<td>W290017</td>
			<td>Phosphoric acid solution, H3PO4, 85 wt. % concentration in water, Mr 98,00</td>
		</tr>
		<tr>
			<td>Sodium chloride physiological solution</td>
			<td>Sigma-Aldrich</td>
			<td>52455</td>
			<td>Tablets of BioUltra NaCl physiological solution; 1 tablet in 1000 mL of water yields 0.9 % NaCl</td>
		</tr>
		<tr>
			<td>Sodium hydroxide, NaOH</td>
			<td>Penta s.r.o.</td>
			<td>40216</td>
			<td>Sodium hydroxide, NaOH, 97,0 %, Mr 40,00</td>
		</tr>
		<tr>
			<td>AmiFarm (Cremor base-A)</td>
			<td>Fagron</td>
			<td>608425</td>
			<td>Hydrophilic oil in water (O/W) cream base; the composition: aqua, paraffin, paraffin liquid, cetearyl alkohol, Laureth 4, sodium hydroxide, carbomer, methylparaben, propylparaben.</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>IKA EUROSTAR POWER control-visc stirrers</td>
			<td>IKA-labortechnik</td>
			<td>Z404020</td>
			<td>Digital laboratory stirrer, for tasks up to the high viscosity range, 230V, 1/cs</td>
		</tr>
		<tr>
			<td>IKA Propeller stirrer, 3-bladed</td>
			<td>IKA-labortechnik</td>
			<td>R 1381</td>
			<td>Propeller stirrer, 3-bladed, stirrer &#216;: 45 mm, shaft &#216;: 8 mm, shaft length: 350 mm</td>
		</tr>
		<tr>
			<td>Dialysis tubing closures</td>
			<td>Sigma-Aldrich</td>
			<td>Z371017-10EA</td>
			<td>Dialysis tubing closures, red, size 110 mm</td>
		</tr>
		<tr>
			<td>Dialysis tubing cellulose membrane</td>
			<td>Sigma-Aldrich</td>
			<td>D9402-100FT</td>
			<td>Dialysis tubing cellulose membrane, average flat width 76 mm (3.0 in.)</td>
		</tr>
		<tr>
			<td>DOMO Pot with stailess, LCD</td>
			<td>DOMO Elektronic</td>
			<td>DO42325PC</td>
			<td>Preserving boiler stainless steel, 2000 W, 27-L container (diameter 37 cm, height 30 cm), temperature control 30-100 &#176; C, operation LCD display</td>
		</tr>
		<tr>
			<td>Hettich zentrifugen Universal 32</td>
			<td>Gemini bv</td>
			<td>2770 GS1R</td>
			<td>Mid bench centrifuge, speed 18000 rpm</td>
		</tr>
		<tr>
			<td>LT 3 shaking device</td>
			<td>Fischer Scientific</td>
			<td>6470.0002</td>
			<td>Orbital shaking device</td>
		</tr>
		<tr>
			<td>KERN 440-47N</td>
			<td>Kern</td>
			<td>440-47N</td>
			<td>Laboratory balance</td>
		</tr>
		<tr>
			<td>KERN 770</td>
			<td>Kern</td>
			<td>770 -N</td>
			<td>Laboratory analytical balance</td>
		</tr>
		<tr>
			<td>VENTICELL 222 - Komfort</td>
			<td>BMT, MMM Group</td>
			<td>C 131749</td>
			<td>Drying oven, temperature control 30-100 &#176; C, air circulation control</td>
		</tr>
		<tr>
			<td>Vacucell 55 - EVO</td>
			<td>BMT, MMM Group</td>
			<td>B 050328</td>
			<td>Vacuum drying oven, temperature control 30-100 &#176; C</td>
		</tr>
		<tr>
			<td>PULVERISETTE 19</td>
			<td>Fritsch</td>
			<td>19.1030.00</td>
			<td>Universal cutting mill, rotor with V-cutting edges and fixed knives</td>
		</tr>
		<tr>
			<td>Multi Probe Adapter System MPA 5</td>
			<td>Courage &#38; Kazaka Electronic</td>
			<td>10225237</td>
			<td>MPA 5 Station - equipment for measurement hydratation, TEWL and pH</td>
		</tr>
		<tr>
			<td>Skin pH-meter PH 905 probe</td>
			<td>Courage &#38; Kazaka Electronic</td>
			<td></td>
			<td>Probe to specifically measure the pH on the skin surface or the scalp</td>
		</tr>
		<tr>
			<td>Corneometer CM 825 probe</td>
			<td>Courage &#38; Kazaka Electronic</td>
			<td></td>
			<td>Probe to determine the hydration level of the skin surface (Stratum corneum).</td>
		</tr>
		<tr>
			<td>Tewameter TM 300</td>
			<td>Courage &#38; Kazaka Electronic</td>
			<td></td>
			<td>Probe for the assessment of the transepidermal water loss (TEWL)</td>
		</tr>
		<tr>
			<td>Heidolph RZR 2020</td>
			<td>Heidolph</td>
			<td>13-225-007-03-1</td>
			<td>Overhead stirrer, mechanical speed setting and stepless transmission; speed range 40-2000 rpm</td>
		</tr>
		<tr>
			<td>Heidolph mechanical stirrer BR 10</td>
			<td>Heidolph</td>
			<td>Z336688-1EA</td>
			<td>Blade impeller crossed stirrer</td>
		</tr>
		<tr>
			<td>Fagor FS 12</td>
			<td>Fagor</td>
			<td>BTT-138</td>
			<td>Laboratory refrigerator with freezer space</td>
		</tr>
		<tr>
			<td>WTW bench pH/mV meter</td>
			<td>WTW</td>
			<td>Z313165</td>
			<td>High-performance bench pH and pH/conductivity meters for routine and high precision laboratory measurements in research or quality control laboratories</td>
		</tr>
		<tr>
			<td>Container</td>
			<td>RPC Superfos</td>
			<td></td>
			<td>13-L plastic bucket, diameter 26 cm, height 26 cm</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Software</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Microsoft Office 2010</td>
			<td>Microsoft</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>C+K software</td>
			<td colspan="2">Courage and Khazaka Electronic GmbH</td>
			<td>MPA 5 station operating software</td>
		</tr>
	</tbody>
</table>

preparation of keratin hydrolysate from chicken feathers and its application in cosmetics

ケラチン加水分解物 (KHs) ヘア化粧品の確立された標準的なコンポーネントであります。KH の保湿効果を理解することは、スキンケア化粧品で有利です。議定書の目標は: (1) アルカリ酵素加水分解による KH に鶏の羽根を処理し、透析による浄化、皮膚の水和が増大し、減少によって皮膚バリア機能を向上させる軟膏ベース (OB) に瀬を追加をテストするには (2)経皮水分喪失 (TEWL)。アルカリ酵素加水分解中に羽が高温アルカリ環境ではし、タンパク質分解酵素で加水分解、温和な条件下で培養しました。KH の溶液を透析、真空乾燥、微粉末に粉砕します。水中油型エマルション (O/W) 2 を含むから構成される化粧品処方、4、および 6 の重量 % (OB の重量に基づいて) KH の用意しています。10 人の男性に KH の保湿のプロパティをテストが行われ、1、2、3、4、24、および 48 h. 試験製剤の間隔で 10 人の女性に脱脂の掌側の前腕のサイトで広がっています。角質層 (SC) の肌の水分補給は最も世界的に使用される、簡単な方法の一つである皮膚の静電容量を測定することによって評価されます。TEWL は、定義された領域と皮膚から期間ごと運ばれる水の量の測定に基づいています。両方のメソッドは完全に非侵襲的です。KH は、優れた閉塞;OB に KH のさらに、によってそれをもたらすアプリケーション後 TEWL を 30% 削減。SC; に表皮の下の層から水をバインドと、KH は、保湿剤としても機能します。OB で最適な KH 足し算を男性と 22% で水和の 19% の上昇に女性の上昇が発生します。

ここに示す手順にしたがって調製された KH (図 2参照) は高蛋白質含有量が付いている水に容易に溶ける色で黄色 (無機固体を表す < 2.0%); KH の 1.0% の解決の pH は 5.3 との要件を満たす化粧品グレードの加水分解物。50 g の原料から KH の収量は約 30% であります。KH の分子量分布は SDS-PAGE によって決定され、図 3に示します。</...

Watch this Scientific Journal Video about Preparation of Keratin Hydrolysate from Chicken Feathers and Its Application in Cosmetics at JoVE.com

Preparation of Keratin Hydrolysate from Chicken Feathers and Its Application in Cosmetics

鶏の羽根や化粧品への応用からケラチンを加水分解物の準備

プロトコルの目的はアルカリ酵素加水分解による羽毛ケラチン加水分解物を準備して化粧品軟膏ベースにケラチンを加水分解物が皮膚バリア機能を向上させる追加するかどうかをテストするのには (ハイドレーションを高めると経皮水分喪失を減少)。検査は、男性と女性のボランティア。

Keratin hydrolysates (KHs) are established standard components in hair cosmetics. Understanding the moisturizing effects of KH is advantageous for ...

preparation-keratin-hydrolysate-from-chicken-feathers-its-application

Research

JoVE Journal

Biochemistry

19.3K ビュー.  Tomas Bata University in Zlín. プロトコルの目的はアルカリ酵素加水分解による羽毛ケラチン加水分解物を準備して化粧品軟膏ベースにケラチンを加水分解物が皮膚バリア機能を向上させる追加するかどうかをテストするのには (ハイドレーションを高めると経皮水分喪失を減少)。検査は、男性と女性のボランティア。

ビデオ: Preparation of Keratin Hydrolysate from Chicken Feathers and Its Application in Cosmetics

Preparation and Delivery of Protein Microcrystals in Lipidic Cubic Phase for Serial Femtosecond Crystallography

Membrane proteins (MPs) are essential components of cellular membranes and primary drug targets. Rational drug design relies on precise structural information, typically obtained by crystallography; however MPs are difficult to crystallize. Recent progress in MP structural determination has benefited greatly from the development of lipidic cubic phase (LCP) crystallization methods, which typically yield well-diffracting, but often small crystals that suffer from radiation damage during traditional crystallographic data collection at synchrotron sources. The development of new-generation X-ray free-electron laser (XFEL) sources that produce extremely bright femtosecond pulses has enabled room temperature data collection from microcrystals with no or negligible radiation damage. Our recent efforts in combining LCP technology with serial femtosecond crystallography (LCP-SFX) have resulted in high-resolution structures of several human G protein-coupled receptors, which represent a notoriously difficult target for structure determination. In the LCP-SFX technique, LCP is recruited as a matrix for both growth and delivery of MP microcrystals to the intersection of the injector stream with an XFEL beam for crystallographic data collection. It has been demonstrated that LCP-SFX can substantially improve the diffraction resolution when only sub-10 &#181;m crystals are available, or when the use of smaller crystals at room temperature can overcome various problems associated with larger cryocooled crystals, such as accumulation of defects, high mosaicity and cryocooling artifacts. Future advancements in X-ray sources and detector technologies should make serial crystallography highly attractive and practicable for implementation not only at XFELs, but also at more accessible synchrotron beamlines. Here we present detailed visual protocols for the preparation, characterization and delivery of microcrystals in LCP for serial crystallography experiments. These protocols include methods for conducting crystallization experiments in syringes, detecting and characterizing the crystal samples, optimizing crystal density, loading microcrystal laden LCP into the injector device and delivering the sample to the beam for data collection.

We describe procedures for the preparation and delivery of membrane protein microcrystals in lipidic cubic phase for serial crystallography at X-ray free-electron lasers and synchrotron sources. These protocols can also be applied for incorporation and delivery of soluble protein microcrystals, leading to substantially reduced sample consumption compared to liquid injection.

準備とシリアルフェムト秒結晶学のための脂質立方相におけるタンパク質微結晶の配達

Membrane proteins (MPs) are essential components of cellular membranes and primary drug targets. Rational drug design relies on precise structural ...

生化学

Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase

Activity-based protein profiling (ABPP) is a method for the identification of an enzyme of interest in a complex proteome through the use of a chemical probe that targets the enzyme&#39;s active sites. A reporter tag introduced into the probe allows for the detection of the labeled enzyme by in-gel fluorescence scanning, protein blot, fluorescence microscopy, or liquid chromatography-mass spectrometry. Here, we describe the preparation and use of the compound ARN14686, a click chemistry activity-based probe (CC-ABP) that selectively recognizes the enzyme N-acylethanolamine acid amidase (NAAA). NAAA is a cysteine hydrolase that promotes inflammation by deactivating endogenous peroxisome proliferator-activated receptor (PPAR)-alpha agonists such as palmitoylethanolamide (PEA) and oleoylethanolamide (OEA). NAAA is synthesized as an inactive full-length proenzyme, which is activated by autoproteolysis in the acidic pH of the lysosome. Localization studies have shown that NAAA is predominantly expressed in macrophages and other monocyte-derived cells, as well as in B-lymphocytes. We provide examples of how ARN14686 can be used to detect and quantify active NAAA ex vivo in rodent tissues by protein blot and fluorescence microscopy.

Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase

Here, we describe the preparation and use of an activity-based probe (ARN14686, undec-10-ynyl-N-[(3S)-2-oxoazetidin-3-yl]carbamate) that allows for the detection and quantification of the active form of the proinflammatory enzyme N-acylethanolamine acid amidase (NAAA), both in vitro and ex vivo.

調製と<em&gt;インビボ</em&gt;アクティビティベースのプローブの使用のために<em&gt; N</em&gt; -acylethanolamine酸アミダーゼ

Activity-based protein profiling (ABPP) is a method for the identification of an enzyme of interest in a complex proteome through the use of a chemical ...

Preparation of Giant Vesicles Encapsulating Microspheres by Centrifugation of a Water-in-oil Emulsion

The constructive biology and the synthetic biology approach to creating artificial life involve the bottom-up assembly of biological or nonbiological materials. Such approaches have received considerable attention in research on the boundary between living and nonliving matter and have been used to construct artificial cells over the past two decades. In particular, Giant Vesicles (GVs) have often been used as artificial cell membranes. In this paper, we describe the preparation of GVs encapsulating highly packed microspheres as a model of cells containing highly condensed biomolecules. The GVs were prepared by means of a simple water-in-oil emulsion centrifugation method. Specifically, a homogenizer was used to emulsify an aqueous solution containing the materials to be encapsulated and an oil containing dissolved phospholipids, and the resulting emulsion was layered carefully on the surface of another aqueous solution. The layered system was then centrifuged to generate the GVs. This powerful method was used to encapsulate materials ranging from small molecules to microspheres.

Giant vesicles containing highly packed micrometer-sized components are useful cell models. The water-in-oil emulsion centrifugation method is a simple, powerful tool for the preparation of giant vesicles with encapsulated materials.

油中水型エマルションの遠心分離により巨大ベシクルカプセル化微小球の調製

The constructive biology and the synthetic biology approach to creating artificial life involve the bottom-up assembly of biological or nonbiological ...

Sample Preparation for Mass Spectrometry-based Identification of RNA-binding Regions

Noncoding RNAs play important roles in several nuclear processes, including regulating gene expression, chromatin structure, and DNA repair. In most cases, the action of noncoding RNAs is mediated by proteins whose functions are in turn regulated by these interactions with noncoding RNAs. Consistent with this, a growing number of proteins involved in nuclear functions have been reported to bind RNA and in a few cases the RNA-binding regions of these proteins have been mapped, often through laborious, candidate-based methods.
Here, we report a detailed protocol to perform a high-throughput, proteome-wide unbiased identification of RNA-binding proteins and their RNA-binding regions. The methodology relies on the incorporation of a photoreactive uridine analog in the cellular RNA, followed by UV-mediated protein-RNA crosslinking, and mass spectrometry analyses to reveal RNA-crosslinked peptides within the proteome. Although we describe the procedure for mouse embryonic stem cells, the protocol should be easily adapted to a variety of cultured cells.

We describe a protocol to identify RNA-binding proteins and map their RNA-binding regions in live cells using UV-mediated photocrosslinking and mass spectrometry.

質量分析を用いた識別 RNA 結合領域のための試料作製

Noncoding RNAs play important roles in several nuclear processes, including regulating gene expression, chromatin structure, and DNA repair. In most ...

Isolation and Respiratory Measurements of Mitochondria from Arabidopsis thaliana

Mitochondria are essential organelles involved in numerous metabolic pathways in plants, most notably the production of adenosine triphosphate (ATP) from the oxidation of reduced compounds such as nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FADH2). The complete annotation of the Arabidopsis thaliana genome has established it as the most widely used plant model system, and thus the need to purify mitochondria from a variety of organs (leaf, root, or flower) is necessary to fully utilize the tools that are now available for Arabidopsis to study mitochondrial biology. Mitochondria are isolated by homogenization of the tissue using a variety of approaches, followed by a series of differential centrifugation steps producing a crude mitochondrial pellet that is further purified using continuous colloidal density gradient centrifugation. The colloidal density material is subsequently removed by multiple centrifugation steps. Starting from 100 g of fresh leaf tissue, 2 - 3 mg of mitochondria can be routinely obtained. Respiratory experiments on these mitochondria display typical rates of 100 - 250 nmol O2 min-1 mg total mitochondrial protein-1 (NADH-dependent rate) with the ability to use various substrates and inhibitors to determine which substrates are being oxidized and the capacity of the alternative and cytochrome terminal oxidases. This protocol describes an isolation method of mitochondria from Arabidopsis thaliana leaves using continuous colloidal density gradients and an efficient respiratory measurements of purified plant mitochondria.

Isolation and Respiratory Measurements of Mitochondria from Arabidopsis thaliana

As mitochondria are only a small percentage of the plant cell, they need to be purified for a range of studies. Mitochondria can be isolated from a variety of plant organs by homogenization, followed by differential and density gradient centrifugation to obtain a highly purified mitochondrial fraction.

分離とシロイヌナズナ由来のミトコンドリアの呼吸測定

Mitochondria are essential organelles involved in numerous metabolic pathways in plants, most notably the production of adenosine triphosphate (ATP) from ...

Genetic Incorporation of Biosynthesized L-dihydroxyphenylalanine (DOPA) and Its Application to Protein Conjugation

L-dihydroxyphenylalanine (DOPA) is an amino acid found in the biosynthesis of catecholamines in animals and plants. Because of its particular biochemical properties, the amino acid has multiple uses in biochemical applications. This report describes a protocol for the genetic incorporation of biosynthesized DOPA and its application to protein conjugation. DOPA is biosynthesized by a tyrosine phenol-lyase (TPL) from catechol, pyruvate, and ammonia, and the amino acid is directly incorporated into proteins by the genetic incorporation method using an evolved aminoacyl-tRNA and aminoacyl-tRNA synthetase pair. This direct incorporation system efficiently incorporates DOPA with little incorporation of other natural amino acids and with better protein yield than the previous genetic incorporation system for DOPA. Protein conjugation with DOPA-containing proteins is efficient and site-specific and shows its usefulness for various applications. This protocol provides protein scientists with detailed procedures for the efficient biosynthesis of mutant proteins containing DOPA at desired sites and their conjugation for industrial and pharmaceutical applications.

Genetic Incorporation of Biosynthesized L-dihydroxyphenylalanine DOPA and Its Application to Protein Conjugation

Here, we present a protocol for the genetic incorporation of L-dihydroxyphenylalanine biosynthesized from simple starting materials and its application to protein conjugation.

遺伝的定款生合成 L-ジヒドロキシフェニルアラニン (DOPA) とタンパク質接合への応用

L-dihydroxyphenylalanine (DOPA) is an amino acid found in the biosynthesis of catecholamines in animals and plants. Because of its particular biochemical ...

Isolation of Lipoprotein Particles from Chicken Egg Yolk for the Study of Bacterial Pathogen Fatty Acid Incorporation into Membrane Phospholipids

Staphylococcus aureus and other Gram-positive pathogens incorporate fatty acids from the environment into membrane phospholipids. During infection, the majority of exogenous fatty acids are present within host lipoprotein particles. Uncertainty remains as to the reservoirs of host fatty acids and the mechanisms by which bacteria extract fatty acids from the lipoprotein particles. In this work, we describe protocols for enrichment of low-density lipoprotein (LDL) particles from chicken egg yolk and determining whether LDLs serve as fatty acid reservoirs for S. aureus. This method exploits unbiased lipidomic analysis and chicken LDLs, an effective and economical model for the exploration of interactions between LDLs and bacteria. The analysis of S. aureus integration of exogenous fatty acids from LDLs is performed using high-resolution/accurate mass spectrometry and tandem mass spectrometry, enabling the characterization of the fatty acid composition of the bacterial membrane and unbiased identification of novel combinations of fatty acids that arise in bacterial membrane lipids upon exposure to LDLs. These advanced mass spectrometry techniques offer an unparalleled perspective of fatty acid incorporation by revealing the specific exogenous fatty acids incorporated into the phospholipids. The methods outlined here are adaptable to the study of other bacterial pathogens and alternative sources of complex fatty acids.

This method provides a framework for studying incorporation of exogenous fatty acids from complex host sources into bacterial membranes, particularly Staphylococcus aureus. To achieve this, protocols for the enrichment of lipoprotein particles from chicken egg yolk and subsequent fatty acid profiling of bacterial phospholipids utilizing mass spectrometry are described.

細菌病原体脂肪酸を膜リン脂質に取り込む研究のための鶏卵黄からのリポタンパク質粒子の単離

Staphylococcus aureus and other Gram-positive pathogens incorporate fatty acids from the environment into membrane phospholipids. During infection, the ...

Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription

A transcription factor&#160;(TF) is a protein that regulates gene expression by interacting with the RNA polymerase, another TF, and/or template DNA. GrgA is a novel transcription activator found specifically in the obligate intracellular bacterial pathogen Chlamydia. Protein pulldown assays using affinity beads have revealed that GrgA binds two &#963; factors, namely &#963;66 and &#963;28, which recognize different sets of promoters for genes whose products are differentially required at developmental stages. We have used BLI to confirm and further characterize the interactions. BLI demonstrates several advantages over pulldown: 1) It reveals real-time association and dissociation between binding partners, 2) It generates quantitative kinetic parameters, and 3) It can detect bindings that pulldown assays often fail to detect. These characteristics have enabled us to deduce the physiological roles of GrgA in gene expression regulation in Chlamydia, and possible detailed interaction mechanisms. We envision that this relatively affordable technology can be extremely useful for studying transcription and other biological processes.

Application of Biolayer Interferometry BLI for Studying Protein-Protein Interactions in Transcription

Interactions of transcription factors (TFs) with the RNA polymerase are usually studied using pulldown assays. We apply a Biolayer Interferometry (BLI) technology to characterize the interaction of GrgA with the chlamydial RNA polymerase. Compared to pulldown assays, BLI detects real-time association and dissociation, offers higher sensitivity, and is highly quantitative.

転写におけるタンパク質とタンパク質の相互作用を研究するための生体層干渉計(BLI)の応用

A transcription factor&#160;(TF) is a protein that regulates gene expression by interacting with the RNA polymerase, another TF, and/or template DNA. GrgA ...

Preparation of High-Quality Fermented Fish Product

This protocol provides a method for preparation of industrialized fermented fish product with sturgeon (Aquilaria sinensis) meat product. The procedures were: (1) pretreatment of farmed sturgeon including decapitation, evisceration, skinning-off, cleaning and cutting; (2) marinating fish cubes in 6-12% (w/v) salt solution (1:1, fish cube mass to solution volume); (3) drying fish cubes to a water content of 50-60% by hot air (40-60 &#176;C) or by vacuum; (4) fermentation involving inoculating fish cubes with 0.4-1.6% (w/w) S. cerevisiae in flavor solution to fish cubes and fermenting at 25-35 &#176;C for 6-10 h; (5) sealing fish cubes in vacuum packages with marinating and fermenting solutions; (6) sterilizing at 115-121 &#176;C for 10-20 min. The sturgeon meat product prepared by this method has delicious taste which is mellow and thick, has various types and large amounts of volatile flavor compounds such as alcohols and esters which could mask musty and unpleasant odor from fish, has moderate salt content but good texture properties such as high springiness, gumminess and chewiness, and has bright russet color and attractive appearance. This new technique could also be applied in the processing of other fish to provide convenient fish snack foods which could be stored at room temperature. It is appropriate for both marine and freshwater fish.

The goal of the protocol is to provide an industrialized fish fermentation technique based on inoculation of Saccharomyces cerevisiae.

高品質発酵魚製品の調製

This protocol provides a method for preparation of industrialized fermented fish product with sturgeon (Aquilaria sinensis) meat product. The procedures ...

JUMPn: A Streamlined Application for Protein Co-Expression Clustering and Network Analysis in Proteomics

With recent advances in mass spectrometry-based proteomics technologies, deep profiling of hundreds of proteomes has become increasingly feasible. However, deriving biological insights from such valuable datasets is challenging. Here we introduce a systems biology-based software JUMPn, and its associated protocol to organize the proteome into protein co-expression clusters across samples and protein-protein interaction (PPI) networks connected by modules (e.g., protein complexes). Using the R/Shiny platform, the JUMPn software streamlines the analysis of co-expression clustering, pathway enrichment, and PPI module detection, with integrated data visualization and a user-friendly interface. The main steps of the protocol include installation of the JUMPn software, the definition of differentially expressed proteins or the (dys)regulated proteome, determination of meaningful co-expression clusters and PPI modules, and result visualization. While the protocol is demonstrated using an isobaric labeling-based proteome profile, JUMPn is generally applicable to a wide range of quantitative datasets (e.g., label-free proteomics). The JUMPn software and protocol thus provide a powerful tool to facilitate biological interpretation in quantitative proteomics.

We present a systems biology tool JUMPn to perform and visualize network analysis for quantitative proteomics data, with a detailed protocol including data pre-processing, co-expression clustering, pathway enrichment, and protein-protein interaction network analysis.