著者はチューレーン大学細胞部と原稿を編集の彼の援助のための分子生物学の博士ロバート dotson さんと彼の援助のため化学のチューレーン大学科の博士ジェームズ ・ ボリンジャーの支援を認める質量分析のプロトコル設計。さらに著者は、チューレーン大学講座細胞・分子生物学化学のチューレーン大学の部門のサポートと空間や設備の使用を認めます。この作品は、チューレーン大学理学と工学から c. アンダーソンにタバコ製品規制科学研究奨学金によって支えられました。

1. デバイスの組立
<ol><li>Buchner フラスコ (図 1#4) 鋼のリングに立つし、乾燥剤として塩化カルシウムの 50 g でそれを充填して真空トラップを作成 100 mL を保護します。ゴム貫通穴栓付けフラスコを密封、パラフィン フィルムとストッパーの接合部をラップし、スルーホール ピペットを実行します。</li>
	<li>ビニール チューブを使用すると、t 字路のホース コネクタ ストッパーから延びるピペットを接続します。</li>
	<li>ビニール チューブを使用すると、互いに 2 つインピン (図 1#2 & #3) を接続し、2 番目のインピンの出力を t 字のホース コネクタに接続します。</li>
	<li>ビニール チューブを使用して、吸入ポート (図 1#1) として最初のインピンの入力ポートに接続します。</li>
	<li>ビニール チューブを使用して空気レギュレータ (図 1#7) の入力ポートに Buchner フラスコのサイドアームを接続し、真空ポンプに空気レギュレータの出口ポート。</li>
	<li>図 2 aの回路図によると回路を組み立てます。</li>
	<li>シリアル アダプターと製造元のソフトウェアを使って BS1 マイクロ コント ローラー (図 1#6) に PBASIC プログラム SVL.bs1 (図 2 b、また https://github.com/ChastainAnderson/SVL で入手可能) をアップロードします。</li>
	<li>リングの場所、510 スレッド ベース (図 1 #8) スタンド クランプです。</li>
	<li>ビニール チューブを使用すると、t 字路のホース コネクタの自由端にソレノイド バルブ (図 1#5) に接続します。 注: デバイスする必要があります完全な操作の準備ができて、タイトな空気が、ホースク ランプおよび必要に応じて真空グリースを適用ことを確認するすべての関節をチェックします。</li>
</ol><img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56709/56709fig2.jpg"> 図 2: 電気回路図と PBASIC コード。図 2 aは両方通常開電磁弁をアクティブ化に必要な電気回路を組み立てるための電気回路図を表示し、ボタンの加熱コイルは、電子タバコを活性化 (、510 をスレッド電子タバコ タンク基本)。加熱コイル (p: 電源の電気的パラメーターR: 抵抗;i: 電流) 投影され、デジタルマルチメータ ポスト アセンブリを実証する必要があります。図 2 bには、図 2 a (https://github.com/ChastainAnderson/SVL も入手可能) に回路を制御するために必要な PBASIC タイミングのプログラムが表示されます。SVT ・ IPT (#5 & #6) タイミング定数は、ms の単位と活性化時間は 2 秒と 28 のダウンタイムを提供するために設定されて s.<a href="//cloudflare.jove.com/files/ftp_upload/56709/56709fig2large.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>。
2. サンプルの調製及び保存
<ol><li>室温で気密のポリ袋で暗闇の中ですべて未開封従来と電子タバコのサンプルを格納します。</li>
	<li>開くと、余分な水分を吸収する紙タオルで 4 ° C で、気密のポリ袋でサンプルを格納します。</li>
	<li>前 ~ 60% 湿度室温ヒュミドールのすべてのサンプルを平衡させ、使用前に少なくとも 30 分間。</li>
</ol>3. 一般的なタバコ煙/電子タバコ エアロゾル抽出装置の操作
<ol><li>各電子タバコ cartomizer/タンク前-vaping 分析用天秤を使用して量を決定します。プレ/ポスト ポスト vaping 重量の差は、適切な投与量を決定する使用されます。 注: 3R4F 参照タバコは、ニコチン 0.7 mg 含まれていると見なされます、商業たばこブランドのニコチン含有量は、従来の分析法13によって定めることが。</li>
	<li>サンプル アプリケーション 1 の血管内皮細胞培養培地の 4.3 ml プライマリ インピンの貯水池を入力します。サンプル アプリケーション 2、アセトン 5 mL でタンクを埋めます。</li>
	<li>
電子タバコや抽出のためタバコの従来を準備します。
	<ol>
<li>従来のタバコを使用して、フィルター周りにクリアのテープの一部を適用、シガレット ペーパーがフィルターを結合容易に目に見える印を置きます。</li>
		<li>電子タバコのようなタバコを使用している場合も充電がバッテリーに堅くねじで締まる、cartomizer いることを確認します。</li>
		<li>電子タバコ タンクを使用する場合は、電子タバコの液体の適切な量がタンクに読み込まれているかどうかを確認し、510 スレッド ベースの上にタンクのネジします。</li>
	</ol>
</li>
	<li>吸入ポート (図 1#1) に従来または電子タバコの先端を挿入し、ホースをクランプで固定します。</li>
	<li>真空ポンプをオンにします。</li>
	<li>プル 1.65 リットル/分 55 mL パフ 2 秒以上を確保するためにフロー メーターを調整します。</li>
	<li>マイクロ コント ローラーをオンにします。従来のタバコを使用している場合は、最初のパフにタバコを点灯します。</li>
	<li>投影まで実行希望濃度 (百万または % 単位で重量/体積) を実現します。</li>
	<li>
分析用天秤を使用して蒸発後各電子タバコ cartomizer/タンクの質量を決定します。この測定測定は、大量消費の合計を決定するステップ 3.1 の撮影に比較します。溶剤量/消費質量濃度を計算します。製品間の正規化に消費されるニコチンのモル濃度を使用します。
	<ol>
<li>十分な質量が消費された場合は、デバイスに電子タバコを返すし、さらに消費します。</li>
		<li>十分なまたは余分な固まりが消費された場合は進みます。</li>
	</ol>
</li>
</ol>4. ろ過とストレージ
<ol><li>エキスの細胞培養に使用する場合は、0.22 μ m ペス シリンジ フィルターを介してフィルターします。</li>
	<li>すぐに抽出物を使用または-80 ° C で保存します。アンダーソン、他のための準備の一部として8、電子タバコ エアロゾルを示した、少なくとも 2 週間安定である、タバコの煙まで 2 年間の安定性は、詐欺師、らによって確立されています。13。</li>
</ol>5. デバイスのクリーニング
<ol><li>それぞれの抽出後チューブを洗い、70% エタノールと脱イオン水を防ぐためにデバイスの貯水池がサンプル間引き継ぐ。</li>
	<li>次の洗浄、簡単にラインの乾燥を支援するために気流を許可する空のデバイスを実行します。</li>
</ol>6. サンプル アプリケーション 1: 中性赤の取り込みセル実行可能性の試金
<ol><li>内皮細胞成長培地の 4.3 mL に上記のように抽出を実行します。</li>
	<li>1 日前、1 x 10 の4セル/好演内皮細胞成長培地の 100 μ L の密度の 96 ウェル プレートにひと臍帯静脈内皮細胞をシードします。</li>
	<li>コントロールとして機能する新鮮な内皮細胞培地のいずれかの 100 μ L または 75 μ L 消費 2 mM ニコチン濃度エキス (1.4 mg を 25 μ l 添加混合内皮細胞成長培地の古い血管内皮細胞の培養液に置き換えてセルを扱う4.3 mL 内皮細胞成長培地にニコチンを消費) 治療として 500 μ M の最終的な集中に。</li>
	<li>両方のタバコの煙、電子たばこエアロゾルのコンポーネントの多くは揮発性、井戸の気密を保つために箔シールを使用します。</li>
	<li>インキュベート プレート 18-24 h 37 ° C および 5% の CO2。</li>
	<li>
ニュートラルレッド染色液を準備します。
	<ol>
<li>バッファー内の塩溶液 10 mL に中立赤の染料 33 mg を溶解して 100 x 中性赤ストック溶液を作成します。</li>
		<li>直前の使用、ニュートラルレッド染色液 x 1 を作成する細胞培養液の原液を 1: 100 x 100 を希釈します。</li>
		<li>ニュートラルレッド染色の使用、および使用する前に、少なくとも 30 分の 37 ° C で液をすぐに孵化させなさい。 メモ: それは潜伏時に析出する結晶の正常です。細胞文化の井戸にこれらの結晶を適用しないようにするには、注意が必要です。必要な場合は、中立赤在庫および染色ソリューションをフィルター処理する.22 μ m フィルターを使用できます。</li>
	</ol>
</li>
	<li>抽出を削除し、ニュートラルレッド染色液適切な定量化のための少なくとも 3 つの空井戸を作成するまあ、超過分の使用あたりの 100 μ L を追加します。</li>
	<li>37 ° C、5% CO2で 2-4 h でプレートを孵化させなさい。</li>
	<li>ニュートラルレッド染色液を削除し、PBS の水没で 3 x を洗ってください。</li>
	<li>中立的な赤を適用解除ソリューション (50% 脱イオン水、49% エタノール, 1% 酢酸) を染色します。</li>
	<li>揺れで部屋の温度で、少なくとも 10 分間、インキュベートします。</li>
	<li>540 で吸光度を読み nm。</li>
	<li>データを分析するには、空白の井戸の平均値を引き算すること、空白の調整コントロールよく価値の平均を正規化します。</li>
</ol>7. サンプル アプリケーション 2: ガスクロマトグラフィー質量分析法
<ol><li>上記としてアセトン 5 mL に抽出を実行します。</li>
	<li>1 ~ 100 の部品の最終的な集中を達成するためにデバイスを実行百万 (e-液体消費/アセトンの量量の重量) サンプルの。</li>
	<li>高精度ガラス製注射器を使用すると、1 μ L を GC-MS によるデバイスのインジェクターに注入します。250 ° C で、1:20 次のオーブン プロトコルで ZB 5 列の結合ガスクロマト/四重極分光システム分割比率注入: 50 ° C で 1 分ランプを 10 ° C/分 140 ° c;300 ° C まで 20 ° C/分と 10 分間保持します。</li>
	<li>エアロゾル成分を識別するために対象のライブラリに結果として得られる質量スペクトルに一致します。</li>
</ol>

<ol>
	<li>World Health Organization. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> WHO Report on the Global Tobacco Epidemic, 2011. , (2011).</li><li>Weaver, S. R., Majeed, B. A., Pechacek, T. F., Nyman, A. L., Gregory, K. R., Eriksen, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+electronic+nicotine+delivery+systems+and+other+tobacco+products+among+USA+adults,+2014:+results+from+a+national+survey.">Use of electronic nicotine delivery systems and other tobacco products among USA adults, 2014: results from a national survey.</a> Int. J. Public Health. 61 (2), 177-188 (2016).</li><li>Singh, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tobacco+Use+Among+Middle+and+High+School+Students+-+United+States,+2011–2015.">Tobacco Use Among Middle and High School Students - United States, 2011–2015.</a> MMWR Morb. Mortal. Wkly. Rep. 65 (14), 361-367 (2016).</li><li>Corey, C. G., Ambrose, B. K., Apelberg, B. J., King, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flavored+Tobacco+Product+Use+Among+Middle+and+High+School+Students--United+States,+2014.">Flavored Tobacco Product Use Among Middle and High School Students--United States, 2014.</a> MMWR Morb. Mortal. Wkly. Rep. 64 (38), 1066-1070 (2015).</li><li>Pisinger, C., D&#248;ssing, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+review+of+health+effects+of+electronic+cigarettes.">A systematic review of health effects of electronic cigarettes.</a> Prev. Med. 69, 248-260 (2014).</li><li>Callahan-Lyon, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electronic+cigarettes:+human+health+effects.">Electronic cigarettes: human health effects.</a> Tob. Control. 23 (Suppl 2), ii36-ii40 (2014).</li><li>Dinakar, C., O'Connor, G. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Health+Effects+of+Electronic+Cigarettes.">The Health Effects of Electronic Cigarettes.</a> N. Engl. J. Med. 375 (14), 1372-1381 (2016).</li><li>Anderson, C., Majeste, A., Hanus, J., Wang, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=E-cigarette+aerosol+exposure+induces+reactive+oxygen+species,+DNA+damage,+and+cell+death+in+vascular+endothelial+cells.">E-cigarette aerosol exposure induces reactive oxygen species, DNA damage, and cell death in vascular endothelial cells.</a> Toxicol. Sci. Off. J. Soc. Toxicol. , (2016).</li><li>U.S. Department of Health and Human Services. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Health Consequences of Smoking: 50 Years of Progress. A Report of the Surgeon General. , (2014).</li><li>Farsalinos, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+the+Cytotoxic+Potential+of+Cigarette+Smoke+and+Electronic+Cigarette+Vapour+Extract+on+Cultured+Myocardial+Cells.">Comparison of the Cytotoxic Potential of Cigarette Smoke and Electronic Cigarette Vapour Extract on Cultured Myocardial Cells.</a> Int. J. Environ. Res. Public. Health. 10 (10), 5146-5162 (2013).</li><li>Schweitzer, K. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+disruptive+proinflammatory+effects+of+nicotine+and+e-cigarette+vapor+exposures.">Endothelial disruptive proinflammatory effects of nicotine and e-cigarette vapor exposures.</a> Am. J. Physiol. - Lung Cell. Mol. Physiol. 309 (2), L175-L187 (2015).</li><li>Putzhammer, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vapours+of+US+and+EU+Market+Leader+Electronic+Cigarette+Brands+and+Liquids+Are+Cytotoxic+for+Human+Vascular+Endothelial+Cells.">Vapours of US and EU Market Leader Electronic Cigarette Brands and Liquids Are Cytotoxic for Human Vascular Endothelial Cells.</a> PLOS ONE. 11 (6), e0157337 (2016).</li><li>Crooks, I., Dillon, D. M., Scott, J. K., Ballantyne, M., Meredith, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+long+term+storage+on+tobacco+smoke+particulate+matter+in+in+vitro+genotoxicity+and+cytotoxicity+assays.">The effect of long term storage on tobacco smoke particulate matter in in vitro genotoxicity and cytotoxicity assays.</a> Regul. Toxicol. Pharmacol. 65 (2), 196-200 (2013).</li><li>Roemer, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mainstream+Smoke+Chemistry+and+in+Vitro+and+In+Vivo+Toxicity+of+the+Reference+Cigarettes+3R4F+and+2R4F.">Mainstream Smoke Chemistry and in Vitro and In Vivo Toxicity of the Reference Cigarettes 3R4F and 2R4F.</a> Beitr. Zur Tab. Contrib. Tob. Res. 25 (1), (2014).</li><li>International Organization for Standards. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> ISO 3088:2012 Routine analytical cigarette smoking machine – Definitions and standard conditions. , (2012).</li><li>World Health Organization. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Standard Operating Procedure for Intense Smoking of Cigarettes. , (2012).</li><li>Brown, C. J., Cheng, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electronic+cigarettes:+product+characterisation+and+design+considerations.">Electronic cigarettes: product characterisation and design considerations.</a> Tob. Control. 23 (Suppl 2), ii4-ii10 (2014).</li><li>Cooperation Centre for Scientific Research Relative to Tobacco. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> CRM No. 81 - Routine Analytical Machine for E-Cigarette Aerosol Generation and Collection - Definitions and Standard Conditions. , (2015).</li><li>Thorne, D., Adamson, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+in+vitro+cigarette+smoke+exposure+systems.">A review of in vitro cigarette smoke exposure systems.</a> Exp. Toxicol. Pathol. 65 (7-8), 1183-1193 (2013).</li><li>Klus, H., Boenke-Nimphius, B., M&#252;ller, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cigarette+Mainstream+Smoke:+The+Evolution+of+Methods+and+Devices+for+Generation,+Exposure+and+Collection.">Cigarette Mainstream Smoke: The Evolution of Methods and Devices for Generation, Exposure and Collection.</a> Beitr. Zur Tab. Contrib. Tob. Res. 27 (4), (2016).</li><li>Baker, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Development+and+Significance+of+Standards+for+Smoking-Machine+Methodology.">The Development and Significance of Standards for Smoking-Machine Methodology.</a> Beitr. Zur Tab. Contrib. Tob. Res. 20 (1), (2014).</li><li>Thorne, D., Crooks, I., Hollings, M., Seymour, A., Meredith, C., Gaca, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mutagenic+assessment+of+an+electronic-cigarette+and+reference+cigarette+smoke+using+the+Ames+assay+in+strains+TA98+and+TA100.">The mutagenic assessment of an electronic-cigarette and reference cigarette smoke using the Ames assay in strains TA98 and TA100.</a> Mutat. Res. Toxicol. Environ. Mutagen. 812, 29-38 (2016).</li><li>Thorne, D., Larard, S., Baxter, A., Meredith, C., Ga&#1195;a, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+comparative+in+vitro+assessment+of+e-cigarette+and+cigarette+smoke+aerosols+using+the+&#947;H2AX+assay+and+applied+dose+measurements.">The comparative in vitro assessment of e-cigarette and cigarette smoke aerosols using the &#947;H2AX assay and applied dose measurements.</a> Toxicol. Lett. 265, 170-178 (2017).</li><li>Herrington, J. S., Myers, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electronic+cigarette+solutions+and+resultant+aerosol+profiles.">Electronic cigarette solutions and resultant aerosol profiles.</a> J. Chromatogr. A. 1418, 192-199 (2015).</li><li>Yu, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electronic+cigarettes+induce+DNA+strand+breaks+and+cell+death+independently+of+nicotine+in+cell+lines.">Electronic cigarettes induce DNA strand breaks and cell death independently of nicotine in cell lines.</a> Oral Oncol. 52, 58-65 (2016).</li><li>Ji, E. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+Electronic+Cigarette+Aerosol+and+Its+Induction+of+Oxidative+Stress+Response+in+Oral+Keratinocytes.">Characterization of Electronic Cigarette Aerosol and Its Induction of Oxidative Stress Response in Oral Keratinocytes.</a> PLOS ONE. 11 (5), e0154447 (2016).</li><li>Morgan, D. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+Reactivity+and+Respiratory+Toxicity+of+the+-Diketone+Flavoring+Agents:+2,3-Butanedione,+2,3-Pentanedione,+and+2,3-Hexanedione.">Chemical Reactivity and Respiratory Toxicity of the -Diketone Flavoring Agents: 2,3-Butanedione, 2,3-Pentanedione, and 2,3-Hexanedione.</a> Toxicol. Pathol. 44 (5), 763-783 (2016).</li><li>Cooperation Centre for Scientific Research Relative to Tobacco. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> CRM No. 84 - Determination of Glycerin, Propylene Glycol, Water, and Nicotine in the Aerosol of E-Cigarettes by Gas Chromatographic Analysis. , (2017).</li></ol>

チューレーン大学によって管理されるタバコ製品規制科学研究員プログラムはアルトリア クライアント サービス規制によって資金を供給します。

このプロトコルの最も重要な要素は、デバイスがきれいなスタートを確実とそれぞれの抽出の完了し、確実にすべてのシールが保持されますので空気の流れが一定です。デバイスが適切にクリーンアップされない場合リスクがあるキャリーの上のサンプルの間です。さらに、装置は長時間時間凝縮したエアロゾルの汚れたまま、乾燥溶媒はシステムをブロックできます。注意してください、?...

医療機関での集中的な努力にもかかわらずタバコ製品の使用タバコ喫煙1に帰因するこれらの死の大半の世界、予防可能な死の主要な原因のまま。2003 年に市場に入り、電子タバコは、タバコ製品のユーザーの間で人気の高まっています。現在、電子タバコはアメリカ大人 (~ 5%)2の間で慣習的なタバコ ・中間 (〜 5.3%) と高校生 (~ 16%)3の中で最も人気のあるニコチン配信システムに最も普及した代わりであります。現在の傾向が続けば場合、は、将来の世代のための慣習的なタバコを交換する電子タバコを期待できます。しかし、電子タバコの使用の健康への影響は不明します。
電子タバコの人気が増加 2013年3,4まで電子タバコに関する研究が本格的に開始されませんでした。それ以来、異なるモデルの数は、彼らの毒性の問題に対処するため採用されています。しかし、多くの研究結果が競合していると、電子タバコは、一般的にある慣習的なタバコより毒性が少ないと思える電子タバコの健康影響に関する現在のコンセンサスは5,を使わない6,7. 私たちの以前の研究は、電子タバコが大幅 DNA 損傷と酸化ストレスと細胞死8 の誘導を引き起こす能力にもかかわらず、従来のタバコよりも血管の内皮細胞に毒性を示します.しかしより多くの研究は、電子タバコの使用の健康への影響についての確固たる結論を描くことができます前に必要です。
慣習的なタバコは予防血管疾患9の主要な原因であるとは、電子タバコの血管の健康リスクの関心の高まりは、10、11,12を使用しています。電子タバコの血管系に及ぼす影響を研究するために私たちの研究室は8の作動マイコン喫煙/vaping デバイス (図 1) を開発しました。このデバイスは、水溶液や有機溶剤のいずれか従来のタバコの煙や電子タバコ エアロゾルの液体抽出物の生成が可能です。調節可能なレギュレーター フローと PBASIC タイミングのプログラムの組み合わせによる空気の流れを制御、デバイスが任意の数のユーザー定義のプロトコルによるとエキスを生成するのに使用できます。ここではアセンブリ、2 つの潜在的なアプリケーションと同様に、このデバイスの操作詳細:体外細胞生存率評価とガスクロマトグラフィー質量分析法。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56709/56709fig1.jpg"> 図 1: 喫煙/Vaping デバイス。たばこ/たばこ電子タバコ (電子セエグ) 構成 (A) のようなタンク電子タバコ構成 (B) で喫煙/vaping デバイスの物理的な組み立ての模式図。コンポーネント キー: 1) 吸入ポート;2) プライマリ コレクション インピン;3) オーバーフロー インピン;4) Buchner フラスコ真空トラップ5) 通常開電磁弁;6) BS1 マイクロ コント ローラー;7) 空気流量制御;8) 510 電子タバコ戦車ベースのスレッド。<a href="//cloudflare.jove.com/files/ftp_upload/56709/56709fig1large.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>。

<table><tbody><tr><td>12 V AC/DC Wall Mount Adaptor</td><td>Digi-Key</td><td>T1099-P5P-ND</td><td></td></tr><tr><td>2.2 Ohm Resistors</td><td>Digi-Key</td><td>A105635-ND</td><td>Used in tandem to generate the 4.4 Ohm resistance in Figure 2A</td></tr><tr><td>330 Ohm Resistors</td><td>Digi-Key</td><td>330QBK-ND</td><td></td></tr><tr><td>510 Threaded Base</td><td>NJoy</td><td>N/A</td><td>Recovered by dismantalling a second generation NJoy electronic cigarette</td></tr><tr><td>Acetic Acid, Glacial</td><td>Sigma-Aldritch</td><td>A6283</td><td></td></tr><tr><td>Acetone (Chromatography Grade)</td><td>Sigma-Aldritch</td><td>34850</td><td></td></tr><tr><td>Basic Stamp Project Board</td><td>Digi-Key</td><td>27112-ND</td><td>This board contains the BS1 Microcontroller, serial adaptor, power switch, and a barrel pin connector for the AC/DC Wall Mount Adaptor</td></tr><tr><td>Basic Stamp USB to Serial Adapter</td><td>Digi-Key</td><td>28030-ND</td><td>An optional component to allow the BS1 serial adaptor to communicate through USB</td></tr><tr><td>Buchner Flask (Vacuum Flask) 250 mL</td><td>VWR</td><td>10545-854</td><td></td></tr><tr><td>Clear Tape</td><td>3M</td><td>S-9783</td><td></td></tr><tr><td>Clear Vinyl Tubing, 3/8&quot; ID</td><td>Watts</td><td>443064</td><td></td></tr><tr><td>EGM-2 Endothelial Cell Culture Medium</td><td>Lonza</td><td>CC-3162</td><td></td></tr><tr><td>Ethanol</td><td>Pharmco-Aaper</td><td>111000200</td><td></td></tr><tr><td>Flow Regulator</td><td>Dwyer</td><td>VFA-23-BV</td><td></td></tr><tr><td>Gas Chromatograph</td><td>Varian</td><td>450-GC</td><td></td></tr><tr><td>Glass Syringe, 10 mL</td><td>Sigma-Aldritch</td><td>Z314552</td><td></td></tr><tr><td>Glass Syringe, 10 &amp;micro;L</td><td>Hamilton</td><td>80300</td><td></td></tr><tr><td>High Vacuum Silicon Grease</td><td>Dow Corning</td><td>146355D</td><td></td></tr><tr><td>Hose Clamp</td><td>Precision Brand</td><td>35125</td><td></td></tr><tr><td>Human Umbilical Vein Endothelial Cells</td><td>ATCC</td><td>PCS-100-013&amp;nbsp;</td><td></td></tr><tr><td>Mass Spectrometer</td><td>Varian</td><td>300-MS</td><td></td></tr><tr><td>Midget Impinger</td><td>Chemglass</td><td>CG-1820-01</td><td></td></tr><tr><td>Neutral Red</td><td>Sigma-Aldritch</td><td>N4638</td><td></td></tr><tr><td>Paraffin Film</td><td>3M</td><td>PM-992</td><td></td></tr><tr><td>Plate Seal Roller</td><td>BioRad</td><td>MSR0001</td><td></td></tr><tr><td>Plate Seal; Foil</td><td>Thermo</td><td>276014</td><td></td></tr><tr><td>Ring Stand 20&quot;</td><td>American Educational Products</td><td>7-G15-A</td><td></td></tr><tr><td>Solenoid Valve (normally open)</td><td>US Solid</td><td>USS2-00081</td><td></td></tr><tr><td>Solid State Relay</td><td>Digi-Key</td><td>CLA279-ND</td><td></td></tr><tr><td>Stand Clamp</td><td>Eisco</td><td>CH0688</td><td></td></tr><tr><td>Syringe Filter, PES, 0.22 um</td><td>Millipore</td><td>SLGP033RS</td><td></td></tr><tr><td>Syringe, 10 mL</td><td>BD Syringe</td><td>309604</td><td></td></tr><tr><td>Through Hole Stopper, Size 6</td><td>VWR</td><td>59581-287</td><td></td></tr><tr><td>Vacuum Pump</td><td>KNF Neuberger</td><td>N86KTP</td><td></td></tr></tbody></table>

a microcontroller operated device for the generation of liquid extracts from conventional cigarette smoke and electronic cigarette aerosol

電子タバコ中、高校生の間で最も人気のあるタバコ製品、大人の間で最も人気のある代替タバコ製品。高品質の電子タバコの使用の結果に再現性のある研究は公衆衛生の懸念を新興国や証拠をクラフト基づく規制政策の理解に不可欠です。論文数の成長は、電子タバコを議論しながら少し一貫性にあるメソッド グループ、結果にほとんどコンセンサス。ここでは、従来のタバコの煙、電子たばこエアロゾルの抽出物を作成するためのプログラマブル試験装置について述べる。このプロトコルのアセンブリおよび当該装置の操作のための指示の詳細し、2 つのサンプル アプリケーションで生成された抽出物の使用方法を示します:体外セル実行可能性の試金およびガスクロマトグラフィー質量分析法。このメソッドは、従来のタバコと電子タバコの間の直接比較を行うためのツールを提供し、電子タバコ研究へアクセスできるエントリ ポイントです。

ひと臍帯静脈の露出の 24 時間以内に従来のタバコの煙のエキス (CSE) または電子タバコ エアロゾルに内皮細胞抽出 (EAE) は重要な (CSE P対コントロール < 0.001; 対 EAE P制御< 0.01;n = 6) セル実行可能性 (図 3 a) の減少。抽出物は、2、2 秒、毎分 55 mL パフの噴き出しプロファイルで生成されて、デバイスによって消費されるニコチンのに基?...

Watch this Scientific Journal Video about A Microcontroller Operated Device for the Generation of Liquid Extracts from Conventional Cigarette Smoke and Electronic Cigarette Aerosol at JoVE.com

A Microcontroller Operated Device for the Generation of Liquid Extracts from Conventional Cigarette Smoke and Electronic Cigarette Aerosol

ここでは、従来のタバコの煙、電子たばこエアロゾルの抽出物を作成するためのプログラマブル試験装置について述べる。このメソッドは、従来のタバコと電子タバコの間の直接比較を行うための便利なツールを提供し、電子タバコ研究へアクセスできるエントリ ポイントです。

マイクロ コント ローラーは従来のタバコの煙、電子たばこエアロゾル液体抽出物の世代のデバイスを運営

Electronic cigarettes are the most popular tobacco product among middle and high schoolers and are the most popular alternative tobacco product among ...

a-microcontroller-operated-device-for-generation-liquid-extracts-from

Universidad de Cartagena UDEC

Research

JoVE Journal

Bioengineering

8.3K ビュー.  Tulane University. ここでは、従来のタバコの煙、電子たばこエアロゾルの抽出物を作成するためのプログラマブル試験装置について述べる。このメソッドは、従来のタバコと電子タバコの間の直接比較を行うための便利なツールを提供し、電子タバコ研究へアクセスできるエントリ ポイントです。

ビデオ: A Microcontroller Operated Device for the Generation of Liquid Extracts from Conventional Cigarette Smoke and Electronic Cigarette Aerosol

Isolation of Mouse Respiratory Epithelial Cells and Exposure to Experimental Cigarette Smoke at Air Liquid Interface

Pulmonary epithelial cells can be isolated from the respiratory tract of mice and cultured at air-liquid interface (ALI) as a model of differentiated respiratory epithelium. A protocol is described for isolating and exposing these cells to mainstream cigarette smoke (CS), in order to study epithelial cell responses to CS exposure. The protocol consists of three parts: the isolation of airway epithelial cells from mouse trachea, the culturing of these cells at air-liquid interface (ALI) as fully differentiated epithelial cells, and the delivery of calibrated mainstream CS to these cells in culture. The ALI culture system allows the culture of respiratory epithelia under conditions that more closely resemble their physiological setting than ordinary liquid culture systems. The study of molecular and lung cellular responses to CS exposure is a critical component of understanding the impact of environmental air pollution on human health. Research findings in this area may ultimately contribute towards understanding the etiology of chronic obstructive pulmonary disease (COPD), and other tobacco-related diseases, which represent major global health problems.

Pulmonary epithelial cells can be isolated from the respiratory tract of mice and cultured at air-liquid interface as a model of differentiated respiratory epithelium. A protocol is described for isolating, culturing and exposing these cells to mainstream cigarette smoke, in order to study molecular responses to this environmental toxin.

エアー液界面におけるマウスの呼吸器上皮細胞と実験たばこの煙への暴露の分離

Pulmonary epithelial cells can be isolated from the respiratory tract of mice and cultured at air-liquid interface (ALI) as a model of differentiated ...

医学

A Protocol for Detecting and Scavenging Gas-phase Free Radicals in Mainstream Cigarette Smoke

Cigarette smoking is associated with human cancers. It has been reported that most of the lung cancer deaths are caused by cigarette smoking 5,6,7,12. Although tobacco tars and related products in the particle phase of cigarette smoke are major causes of carcinogenic and mutagenic related diseases, cigarette smoke contains significant amounts of free radicals that are also considered as an important group of carcinogens9,10. Free radicals attack cell constituents by damaging protein structure, lipids and DNA sequences and increase the risks of developing various types of cancers. Inhaled radicals produce adducts that contribute to many of the negative health effects of tobacco smoke in the lung3. Studies have been conducted to reduce free radicals in cigarette smoke to decrease risks of the smoking-induced damage. It has been reported that haemoglobin and heme-containing compounds could partially scavenge nitric oxide, reactive oxidants and carcinogenic volatile nitrosocompounds of cigarette smoke4. A 'bio-filter' consisted of haemoglobin and activated carbon was used to scavenge the free radicals and to remove up to 90% of the free radicals from cigarette smoke14. However, due to the cost-ineffectiveness, it has not been successfully commercialized. Another study showed good scavenging efficiency of shikonin, a component of Chinese herbal medicine8. In the present study, we report a protocol for introducing common natural antioxidant extracts into the cigarette filter for scavenging gas phase free radicals in cigarette smoke and measurement of the scavenge effect on gas phase free radicals in mainstream cigarette smoke (MCS) using spin-trapping Electron Spin Resonance (ESR) Spectroscopy1,2,14. We showed high scavenging capacity of lycopene and grape seed extract which could point to their future application in cigarette filters. An important advantage of these prospective scavengers is that they can be obtained in large quantities from byproducts of tomato or wine industry respectively11,13

Spin-trapping ESR spectroscopy was used to study the effect of plant antioxidants lycopene, pycnogenol and grape seed extract on scavenging gas-phase free radicals in cigarette smoke.

たばこの主流煙のフリーラジカルを検出し、気相を清掃するためのプロトコル

Cigarette smoking is associated with human cancers. It has been reported that most of the lung cancer deaths are caused by cigarette smoking 5,6,7,12.  ...

生物工学

Bridging the Bio-Electronic Interface with Biofabrication

Advancements in lab-on-a-chip technology promise to revolutionize both research and medicine through lower costs, better sensitivity, portability, and higher throughput. The incorporation of biological components onto biological microelectromechanical systems (bioMEMS) has shown great potential for achieving these goals. Microfabricated electronic chips allow for micrometer-scale features as well as an electrical connection for sensing and actuation. Functional biological components give the system the capacity for specific detection of analytes, enzymatic functions, and whole-cell capabilities. Standard microfabrication processes and bio-analytical techniques have been successfully utilized for decades in the computer and biological industries, respectively. Their combination and interfacing in a lab-on-a-chip environment, however, brings forth new challenges. There is a call for techniques that can build an interface between the electrode and biological component that is mild and is easy to fabricate and pattern.
Biofabrication, described here, is one such approach that has shown great promise for its easy-to-assemble incorporation of biological components with versatility in the on-chip functions that are enabled. Biofabrication uses biological materials and biological mechanisms (self-assembly, enzymatic assembly) for bottom-up hierarchical assembly. While our labs have demonstrated these concepts in many formats 1,2,3, here we demonstrate the assembly process based on electrodeposition followed by multiple applications of signal-based interactions. The assembly process consists of the electrodeposition of biocompatible stimuli-responsive polymer films on electrodes and their subsequent functionalization with biological components such as DNA, enzymes, or live cells 4,5. Electrodeposition takes advantage of the pH gradient created at the surface of a biased electrode from the electrolysis of water 6,7,. Chitosan and alginate are stimuli-responsive biological polymers that can be triggered to self-assemble into hydrogel films in response to imposed electrical signals 8. The thickness of these hydrogels is determined by the extent to which the pH gradient extends from the electrode. This can be modified using varying current densities and deposition times 6,7. This protocol will describe how chitosan films are deposited and functionalized by covalently attaching biological components to the abundant primary amine groups present on the film through either enzymatic or electrochemical methods 9,10. Alginate films and their entrapment of live cells will also be addressed 11. Finally, the utility of biofabrication is demonstrated through examples of signal-based interaction, including chemical-to-electrical, cell-to-cell, and also enzyme-to-cell signal transmission. 
Both the electrodeposition and functionalization can be performed under near-physiological conditions without the need for reagents and thus spare labile biological components from harsh conditions. Additionally, both chitosan and alginate have long been used for biologically-relevant purposes 12,13. Overall, biofabrication, a rapid technique that can be simply performed on a benchtop, can be used for creating micron scale patterns of functional biological components on electrodes and can be used for a variety of lab-on-a-chip applications.

This article describes a biofabrication approach: deposition of stimuli-responsive polysaccharides in the presence of biased electrodes to create biocompatible films which can be functionalized with cells or proteins. We demonstrate a bench-top strategy for the generation of the films as well as their basic uses for creating interactive biofunctionalized surfaces for lab-on-a-chip applications.

Biofabricationとバイオエレクトロニック·インタフェースをブリッジする

Advancements in lab-on-a-chip technology promise to revolutionize both research and medicine through lower costs, better sensitivity, portability, and ...

Generation of Electronic Cigarette Aerosol by a Third-Generation Machine-Vaping Device: Application to Toxicological Studies

Electronic-cigarette (e-cig) devices use heat to produce an inhalable aerosol from a liquid (e-liquid) composed mainly of humectants, nicotine, and flavoring chemicals. The aerosol produced includes fine and ultrafine particles, and potentially nicotine and aldehydes, which can be harmful to human health. E-cig users inhale these aerosols and, with the third-generation of e-cig devices, control design features (resistance and voltage) in addition to the choice of e-liquids, and the puffing profile. These are key factors that can significantly impact the toxicity of the inhaled aerosols. E-cig research, however, is challenging and complex mostly due to the absence of standardized assessments and to the numerous varieties of e-cig models and brands, as well as e-liquid flavors and solvents that are available on the market. These considerations highlight the urgent need to harmonize e-cig research protocols, starting with e-cig aerosol generation and characterization techniques. The current study focuses on this challenge by describing a detailed step-by-step e-cig aerosol generation technique with specific experimental parameters that are thought to be realistic and representative of real-life exposure scenarios. The methodology is divided into four sections: preparation, exposure, post-exposure analysis, plus cleaning and maintenance of the device. Representative results from using two types of e-liquid and various voltages are presented in terms of mass concentration, particle size distribution, chemical composition and cotinine levels in mice. These data demonstrate the versatility of the e-cig exposure system used, aside from its value for toxicological studies, as it allows for a broad range of computer-controlled exposure scenarios, including automated representative vaping topography profiles.

Electronic cigarette (e-cig) users are increasing worldwide. Little, however, is known about the health effects induced by inhaled e-cig aerosols. This article describes an e-cig aerosol generation technique suitable for animal exposures and subsequent toxicological studies. Such protocols are required to establish experimentally reproducible and standardized e-cig exposure systems.

第三世代機 Vaping デバイスによる電子タバコ エアロゾルの生成: 毒性学的研究への応用

Electronic-cigarette (e-cig) devices use heat to produce an inhalable aerosol from a liquid (e-liquid) composed mainly of humectants, nicotine, and ...

化学

Use of Capillary Aerosol Generator in Continuous Production of Controlled Aerosol for Non-Clinical Studies

The capillary aerosol generator (CAG) is operated with the principal of thermal liquid evaporation through heating of e-liquid in the initial phase, followed by nucleation and condensation regulated through a mixture of airflow to generate aerosols, such as in an electronic cigarette (EC). The CAG is particularly useful in generating aerosols of large volumes in a continuous manner, for instances such as in vivo inhalation toxicology studies, where usage of ECs is not feasible. The thermal effects of generating aerosol from the CAG are similar in terms of temperature applied in an EC, thus allowing investigators to assess the vapors of e-liquids at scale and reproducibility. As the operation of the CAG allows users to control critical parameters such as the flow rate of e-liquid, heating temperatures and dilution air flows, it allows investigators to test various e-liquid formulations in a well-controlled device. Properties, such as aerosol particle size, are demonstrated to be regulated with the air flow rate with respect to the e-liquid flow and e-liquid composition. The CAG, however, is limited in assessing common EC-related issues, such as overheating of its elements. We seek to demonstrate that the CAG can generate aerosol that is reproducible and continuous, by assessing the chemical and physical aerosol characteristics with a chosen e-liquid formulation. The protocol describes the operating parameters of liquid flow rate, dilution air-flow rates and operating procedures needing to optimize the aerosol concentration and particle size required for an in vivo toxicology study. Presenting the representative results from the protocol and discussing the challenges and applications of working with a CAG, we demonstrate that CAG can be used in a reproducible fashion. The technology and protocol, that has been developed from prior work, serve as a foundation for future innovations for laboratory-controlled aerosol generation investigations.

The protocol describes the settings and use of a capillary aerosol generator for continuous production of controlled aerosol from a multispecies liquid solution, suitable for steady large-volume aerosol delivery (e.g., in vivo inhalation studies).

非臨床試験のための制御エアロゾルの連続生産におけるキャピラリーエアロゾル発生器の使用

The capillary aerosol generator (CAG) is operated with the principal of thermal liquid evaporation through heating of e-liquid in the initial phase, ...

工学

A Versatile Kit Based on Digital Microfluidics Droplet Actuation for Science Education

This paper describes an educational kit based on digital microfluidics. A protocol for luminol-based chemiluminescence experiment is reported as a specific example. It also has fluorescent imaging capability and closed humidified enclosure based on an ultrasonic atomizer to prevent evaporation. The kit can be assembled within a short period of time and with minimal training in electronics and soldering. The kit allows both undergraduate/graduate students and enthusiasts to obtain hands-on experience in microfluidics in an intuitive way and be trained to gain familiarity with digital microfluidics.

We describe an educational kit that allows users to execute multiple experiments and gain hands-on experience on digital microfluidics.

科学教育のためのデジタルマイクロ流体ドロップレット作動に基づく多目的なキット

This paper describes an educational kit based on digital microfluidics. A protocol for luminol-based chemiluminescence experiment is reported as a ...

Automated Measurement of Pulmonary Emphysema and Small Airway Remodeling in Cigarette Smoke-exposed Mice

COPD is projected to be the third most common cause of mortality world-wide by 2020(1). Animal models of COPD are used to identify molecules that contribute to the disease process and to test the efficacy of novel therapies for COPD. Researchers use a number of models of COPD employing different species including rodents, guinea-pigs, rabbits, and dogs(2). However, the most widely-used model is that in which mice are exposed to cigarette smoke. Mice are an especially useful species in which to model COPD because their genome can readily be manipulated to generate animals that are either deficient in, or over-express individual proteins. Studies of gene-targeted mice that have been exposed to cigarette smoke have provided valuable information about the contributions of individual molecules to different lung pathologies in COPD(3-5). Most studies have focused on pathways involved in emphysema development which contributes to the airflow obstruction that is characteristic of COPD. However, small airway fibrosis also contributes significantly to airflow obstruction in human COPD patients(6), but much less is known about the pathogenesis of this lesion in smoke-exposed animals. To address this knowledge gap, this protocol quantifies both emphysema development and small airway fibrosis in smoke-exposed mice. This protocol exposes mice to CS using a whole-body exposure technique, then measures respiratory mechanics in the mice, inflates the lungs of mice to a standard pressure, and fixes the lungs in formalin. The researcher then stains the lung sections with either Gill&#8217;s stain to measure the mean alveolar chord length (as a readout of emphysema severity) or Masson&#8217;s trichrome stain to measure deposition of extracellular matrix (ECM) proteins around small airways (as a readout of small airway fibrosis). Studies of the effects of molecular pathways on both of these lung pathologies will lead to a better understanding of the pathogenesis of COPD.

The goal of this protocol is to provide automated methods to quantify chronic lung pathologies in a murine model of COPD. The protocol includes exposing mice to cigarette smoke (CS), measuring pulmonary function, inflating the lungs, and using morphometry methods to measure emphysema and small airway remodeling in mice.

肺気腫とタバコ煙曝露マウスにおける末梢気道リモデリングの自動測定

COPD is projected to be the third most common cause of mortality world-wide by 2020(1).  Animal models of COPD are used to identify molecules that ...

Comparing the Effects of Electronic Cigarette Vapor and Cigarette Smoke in a Novel In Vivo Exposure System

Electronic cigarettes (E-cigarettes) are being widely used, and growing in popularity. It is estimated that more than 9 million adults use them regularly. The potential adverse health effects of electronic cigarette vapor (E-vapor) exposure are poorly defined. While several animal models of E-vapor exposure have been developed, few models expose rodents to clinically relevant quantities of nicotine and make direct comparisons to cigarette smoke within the same exposure system. Here, we present a method for constructing and operating an E-vapor chamber and cigarette smoke chamber. The chambers are constructed by outfitting anesthesia chambers with a computer controlled pumping system that delivers consistent amounts of E-vapor or cigarette smoke to rodents. Nicotine exposure is measured indirectly by quantifying pre and post-exposure serum cotinine levels. This exposure system can be modified to accommodate various types of E-cigarettes&#160;and tobacco cigarettes, and can be used to compare the effects of E-vapor&#160;and cigarette smoke in vivo.

Comparing the Effects of Electronic Cigarette Vapor and Cigarette Smoke in a Novel In Vivo Exposure System

This protocol describes a method for exposing rodents to electronic cigarette vapor (E-vapor) and cigarette smoke. Exposure chambers are constructed by modifying anesthesia chambers with an automated pumping system that delivers E-vapor or cigarette smoke to rodents. This system can easily be modified to accommodate many experimental endpoints.

電子たばこの蒸気とたばこの煙が小説に与える影響の比較<em&gt;インビボ</em&gt;露光システム

Electronic cigarettes (E-cigarettes) are being widely used, and growing in popularity. It is estimated that more than 9 million adults use them regularly. ...

Cigarette Smoke Exposure in Mice using a Whole-Body Inhalation System

Close to 14% of adults in the United States were reported to smoke cigarettes in 2018. The effects of cigarette smoke (CS) on lungs and cardiovascular diseases have been widely studied, however, the impact of CS in other tissues and organs such as blood and bone marrow remain incompletely defined. Finding the appropriate system to study the effects of CS in rodents can be prohibitively expensive and require the purchase of commercially available systems. Thus, we set out to build an affordable, reliable, and versatile system to study the pathologic effects of CS in mice. This whole-body inhalation exposure system (WBIS) set-up mimics the breathing and puffing of cigarettes by alternating exposure to CS and clean air. Here we show that this do-it-yourself (DIY) system induces airway inflammation and lung emphysema in mice after 4-months of cigarette smoke exposure. The effects of whole-body inhalation (WBI) of CS on hematopoietic stem and progenitor cells (HSPCs) in the bone marrow using this apparatus are also shown.

This protocol demonstrates the study of the pathophysiologic effects of cigarette smoke (CS) with a whole-body inhalation (WBI) exposure system (WBIS) built in-house. This system can expose animals to CS under controlled repeatable conditions for research of CS-mediated effects on lung emphysema and hematopoiesis.

全身吸入システムを用いたマウスのタバコの煙暴露

Close to 14% of adults in the United States were reported to smoke cigarettes in 2018. The effects of cigarette smoke (CS) on lungs and cardiovascular ...

免疫・感染学

A Technique to Generate and Deliver Drug Aerosols Directly to Mouse Lungs

Source: Phillips, J. E. et al., <a href="https://appjove.unicartagenaproxy.elogim.com/v/55454">Dry Powder and Nebulized Aerosol Inhalation of Pharmaceuticals Delivered to Mice Using a Nose-only Exposure System.</a>&#160;J. Vis. Exp. (2017)
This video demonstrates the generation of drug aerosols and the targeted nose-only delivery of these aerosols to mice using an inhalation unit equipped with an aerosol generator. The mouse is restrained in a nose-only restrainer and connected to the inhalation unit, which delivers the drug directly into the mouse&#39;s nasal cavity. Subsequently, the drug passes through the trachea and enters the lungs.

薬物エアロゾルを生成してマウスの肺に直接送達する技術

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

マイクロコントローラーは従来のタバコの煙、電子たばこエアロゾル液体抽出物の世代のデバイスを運営

要約

さらに動画を探す

この動画の章

エアー液界面におけるマウスの呼吸器上皮細胞と実験たばこの煙への暴露の分離

たばこの主流煙のフリーラジカルを検出し、気相を清掃するためのプロトコル

Biofabricationとバイオエレクトロニック·インタフェースをブリッジする

第三世代機 Vaping デバイスによる電子タバコエアロゾルの生成: 毒性学的研究への応用

非臨床試験のための制御エアロゾルの連続生産におけるキャピラリーエアロゾル発生器の使用

科学教育のためのデジタルマイクロ流体ドロップレット作動に基づく多目的なキット

肺気腫とタバコ煙曝露マウスにおける末梢気道リモデリングの自動測定

電子たばこの蒸気とたばこの煙が小説に与える影響の比較

全身吸入システムを用いたマウスのタバコの煙暴露

薬物エアロゾルを生成してマウスの肺に直接送達する技術

マイクロ コント ローラーは従来のタバコの煙、電子たばこエアロゾル液体抽出物の世代のデバイスを運営

要約

さらに動画を探す

この動画の章

エアー液界面におけるマウスの呼吸器上皮細胞と実験たばこの煙への暴露の分離

たばこの主流煙のフリーラジカルを検出し、気相を清掃するためのプロトコル

Biofabricationとバイオエレクトロニック·インタフェースをブリッジする

第三世代機 Vaping デバイスによる電子タバコ エアロゾルの生成: 毒性学的研究への応用

非臨床試験のための制御エアロゾルの連続生産におけるキャピラリーエアロゾル発生器の使用

科学教育のためのデジタルマイクロ流体ドロップレット作動に基づく多目的なキット

肺気腫とタバコ煙曝露マウスにおける末梢気道リモデリングの自動測定

電子たばこの蒸気とたばこの煙が小説に与える影響の比較

全身吸入システムを用いたマウスのタバコの煙暴露

薬物エアロゾルを生成してマウスの肺に直接送達する技術

マイクロコントローラーは従来のタバコの煙、電子たばこエアロゾル液体抽出物の世代のデバイスを運営

第三世代機 Vaping デバイスによる電子タバコエアロゾルの生成: 毒性学的研究への応用