この研究は、基礎科学研究プログラムによって支えられた (プロジェクト番号: 2016R1A2B4013467) を通じて国立研究財団の韓国 (NRF) 科学省 ICT ・将来計画 2016 によって資金を供給し、韓国基礎科学でもサポートされています。研究プログラム研究所 (プロジェクト番号: C36707)。

1. DMMTAVの合成
<ol><li>
化学準備とモル比混合 DMAVナ2S、H2SO4 注: DMAV: Na2S:H24 = 1:1.6:1.6<ol>
<li>40 mL の脱イオンと N2の DMAVの 5.24 g を溶解-パージ (少なくとも 30 分はパージ) 50 mL 遠心管中の水。</li>
		<li>Na2S·9H2O で 50 mL の脱イオンと N2の 14.41 g を溶解することにより Na2S 試薬を準備-250 mL フラスコに水を削除します。</li>
		<li>準備 H2純水 40 mL に濃硫酸 (96%) の 3.3 mL を追加して4試薬だと N2-50 mL の遠心管中の水をパージします。 注: DMAVの最終的なモル比: Na2S:H2SO4 = 1:1.6:1.6 1,3,4,8,15。</li>
		<li>DMAVソリューション (ステップ 1.1.1) の準備 40 mL を 250 mL のフラスコ (ステップ 1.1.2) に含まれる Na2S 溶液 50 mL に追加します。DMAV 10 mL のパージされた水で管内を洗浄し、250 mL フラスコを同様にこれを追加します。</li>
		<li>ガラス管が装備 3 つ穴ゴム栓付けフラスコを閉じます。ガラスを使用して、N2ガス流入、流出と H2SO4液入口それぞれチューブします。フラスコを閉じた直後にフラスコに N2ガスの流れを許可します。 注: 反応液の表面に飛散することがなく流れにガス圧を維持しなければなりません。</li>
		<li>耐酸性チューブ H2のガラス チューブにその4液入口 H2の準備の 40 mL を追加しに 50 mL 注射器を接続4ソリューション (1.1.3 ステップ)。H2の 40 mL を追加し、4ソリューションでは、ゆっくりと段階的に。 注意:硫酸を追加するには、時に白い煙が生成されます。換気ヒューム フードを使用します。</li>
		<li>定期的に硫酸を加えている間フラスコ内で反応混合物の色の変化を監視します。だから、H2の 4-5 mL の間隔が値下がりしましたを維持4;混合物は、白い曇りのソリューションにする必要があります。 注: 急速な濃硫酸添加による瞬時の黄色い沈殿物があります。</li>
		<li>1.1.4 のステップの始まりから 1 時間立っている反応液を確認します。</li>
	</ol>
</li>
	<li>
DMMTAV抽出液-液抽出法を用いて
	<ol>
<li>1 時間後に、ジエチル エーテルの約 200 mL を含む, 漏斗に反応液を注ぐ。</li>
		<li>5-10 分、活栓を切り替えることによって数回ガスを放出の漏斗を振る。 注: 合成 DMMTAVはジエチル エーテル (0.713 g·mL-1) の上位層に転送されます。 注意:ジエチル エーテルのガスは有害かもしれない換気ヒューム フードを使用します。</li>
		<li>、ビーカーに反応液を収集し、個別にボトルの DMMTAVを含むジエチル エーテルを収集します。同じ, 漏斗に反応液を置き、reshaking の新鮮なジエチル エーテルの約 200 mL を追加します。1.2.2 - 1.2.3 の手順を 3 回繰り返します。</li>
		<li>もう一度、同じ, 漏斗にステップ 1.2.3 から収集したジエチル エーテルを注ぐし、約 100 mL の N2を追加-脱イオン水を削除します。5-10 分のために振るし、N2破棄-純水と純度のためジエチル エーテルの数 mL をパージします。ガラス シャーレ (160 mm の最小内径) と最小の高さ 50 mm で残りのジエチル エーテルを収集します。</li>
		<li>種変換を防ぐために N2雰囲気グローブ ボックスにガラス シャーレを転送します。 注意:パス ボックスのコンセントを通じて真空ポンプに溶媒を抜けないことを確認します。</li>
		<li>Dimethylmonothioarsinate の白色沈殿物まで乾燥 (結晶化 DMMTAV) は、ガラス シャーレに形成されます。 注: プロトコルはここで一時停止することができます。</li>
	</ol>
</li>
	<li>
合成 DMMTAVおよび記憶域の検証
	<ol>
<li>DMMTAVの結晶の白色沈殿物を取るとを測定し、総重量を記録します。 注意:硫化水素ガスを吸入しないように換気の良いヒューム フードやグローブ ボックスを使用します。</li>
		<li>ディゾルブ N2の 50 mL の DMMTAVを結晶化-脱イオン水をパージ、0.2 μ m のシリンジ フィルターを介して黄色の沈殿物をフィルター処理します。</li>
		<li>現在使用されている DMAV合計は DMMTAV、すなわち≈9、649 mg As· に変換されますと仮定L-1。DMMTAV ≈1 mg· に原液を希釈します。L-1と ≈40 µg·エレクトロ スプレー イオン化を用いた検証解析 L-1質量 Spectromtery (MS)、高速液体クロマトグラフィー-ICP-MS、それぞれ。</li>
		<li>DMMTAV m/z 155 ポジティブ イオン モードで、 m/zマイナス イオン モード (表 1) で 153 MS11,16とフラグメントを使用してのm/zを分析します。 注: m/z (表 1) の参照値を参照してください。</li>
		<li>適切な溶離液条件による高速液体クロマトグラフィー-誘導結合プラズマ質量11,16,17原液の DMMTAVのクロマト グラムを分析および確認に記載されている保存時に大きなピークが検出、文学。 注: 合成 DMMTAVの純度は、1.3.5 のステップから解析結果を用いた計算する必要があります。</li>
		<li>酸分解11後 ICP-MS を用いた総砒素濃度を分析し、合成 DMMTAV原液希釈倍率と次式のように純度を使用して真の DMMTAVの濃度を計算します。 濃度 (µg· として分析された合計L-1) ·希釈係数 ·純度 (%) = True DMMTAV DMMTAV原液 (µg· 濃度L-1)</li>
		<li>暗闇のなかでさらに定量的なスペシエーション分析18DMMTAV 4 ° C で原液を保存します。</li>
	</ol>
</li>
</ol>DMDTAVの合成
<ol><li>
化学準備とモル比混合 DMAVナ2S、H2SO4 注: DMAV: Na2S:H24 = 1:7.5:7.5<ol>
<li>40 mL の脱イオン水 50 mL 遠心管中の DMAVの 1.38 g を溶解します。</li>
		<li>ナ2250 mL フラスコ 50 mL の脱イオン水 S·9H2O の 18.01 g を溶解することにより Na2S 試薬を準備します。</li>
		<li>40 mL の脱イオン水に濃硫酸 (96%) の 4 つの mL を追加して4試薬は 50 mL の遠心管に含まれるように H2を準備します。 注: DMAVの最終的なモル比: Na2S:H2SO4 = 1:7.5:7.51,3,4,8,15。</li>
		<li>DMAVソリューション (ステップ 2.1.1) の準備の 40 mL を 250 mL のフラスコ (ステップ 2.1.2) に含まれる Na2S 溶液 50 mL に追加します。DMAV 10 mL の脱イオン水で管内を洗浄し、250 mL フラスコを同様にこれを追加します。</li>
		<li>H2の準備の 40 mL を追加し、4ソリューション (2.1.3 ステップ)、ゆっくりとフラスコに、段階的に。</li>
		<li>定期的に硫酸を加えている間フラスコ内で反応液の色の変化を監視します。だから、H2の 4-5 mL の間隔が値下がりしましたを維持4;混合物は、白/黄色曇りソリューションをする必要があります。 注: 急速な濃硫酸添加により瞬時の黄色い沈殿物があります。 注意:硫酸を追加するには、時に白い煙が生成されます。風通しヒューム フードを使用します。</li>
		<li>カバーなし一晩フラスコ反応液を維持します。 注: プロトコルはここで一時停止することができます。</li>
	</ol>
</li>
	<li>
DMDTAV抽出固相抽出 (SPE) を用いた
	<ol>
<li>一晩立って反応後 C18 シリンジを使用して反応液をトラップするためにシリカ系の SPE の入力フィルターは合成樹脂に DMDTAVです。 注意:換気ヒューム フードを使用します。</li>
		<li>10 mM 酢酸アンモニウム (pH 6.3) を準備するには、1 L の脱イオン水に酢酸アンモニウム 0.77 g を溶解します。10 mM 酢酸アンモニウムの十分な量を溶出吸着 DMDTAVを抽出する C18 シリンジ (ステップ 2.2.1) を介して。ガラス シャーレ (160 mm の最小内径) と最小の高さ 50 mm でフィルター処理されたアンモニウム酢酸を収集します。</li>
		<li>種変換を防ぐために N2雰囲気グローブ ボックスにガラス シャーレを転送します。 注意:パス ボックスのコンセントを通じて真空ポンプに溶剤を抜けないことを確認します。</li>
		<li>Dimethyldithioarsinate の白色沈殿物まで乾燥ガラス シャーレに (結晶化 DMDTAV) を結成します。 注: プロトコルはここで一時停止することができます。</li>
	</ol>
</li>
	<li>
合成 DMDTAVおよび記憶域の検証
	<ol>
<li>結晶 DMDTAV、白色の沈殿物を取るし、総重量は、測定の記録を測定します。 注意:硫化水素ガスを吸入しないように換気のヒューム フードやグローブ ボックスを使用します。</li>
		<li>ディゾルブ N2の 50 mL の DMDTAVを結晶化-パージされた脱イオン水 50 mL にチューブを遠心し、0.2 μ m のシリンジ フィルターを介して任意の沈殿物をフィルター処理します。</li>
		<li>現在使用されている DMAV合計は DMDTAV、すなわち≈2 539 mg As· に変換されると仮定します。L-1。DMDTAV ≈1 mg· に原液を希釈します。L-1と ≈40 µg·それぞれ MS と高速液体クロマトグラフィー-ICP-MS を用いた検証解析 L-1 。</li>
		<li>DMDTAV m/z 171 ポジティブ イオン モードで、 m/zマイナス イオン モード (表 1) で 169 ESI MS11,16とフラグメントを使用してのm/zを分析します。 注: m/z (表 1) の参照値を参照してください。</li>
		<li>適切な溶離液条件による高速液体クロマトグラフィー-誘導結合プラズマ質量11,16,17原液の DMDTAVのクロマト グラムを分析し、「保持時間ピークがあることを確認文学。 注: 合成 DMDTAVの純度は、2.3.5 のステップから解析結果を用いた計算する必要があります。</li>
		<li>酸分解11後 ICP-MS を用いた総砒素濃度を分析し、次の式として希釈倍率と純度を使用して合成の DMDTAV在庫ソリューションで真の DMDTAV濃度を計算します。 濃度 (µg· として分析された合計L-1) ·希釈係数 ·純度 (%) = True DMDTAV DMDTAV原液 (µg· 濃度L-1)</li>
		<li>暗闇のなかでさらに定量的なスペシエーション分析18DMDTAV 4 ° C で原液を保存します。</li>
	</ol>
</li>
</ol>

<ol>
	<li>Suzuki, K. 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=Dimethylthioarsenicals+as+arsenic+metabolites+and+their+chemical+preparation.">Dimethylthioarsenicals as arsenic metabolites and their chemical preparation.</a> Chem. Res. Toxicol. 17, 914-921 (2004).</li><li>Kuroda, 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=Microbial+metabolite+of+dimethylarsinic+acid+is+highly+toxic+and+genotoxic.">Microbial metabolite of dimethylarsinic acid is highly toxic and genotoxic.</a> Toxicol. Appl. Pharmacol. 198, 345-353 (2004).</li><li>Naranmandura, H., Iwata, K., Suzuki, K. T., Ogra, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distribution+and+metabolism+of+four+different+dimethylated+arsenicals+in+hamsters.">Distribution and metabolism of four different dimethylated arsenicals in hamsters.</a> Toxicol. Appl. Pharmacol. 245, 67-75 (2010).</li><li>Naranmandura, 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=Comparative+toxicity+of+arsenic+metabolites+in+human+bladder+cancer+EJ-1+cells.">Comparative toxicity of arsenic metabolites in human bladder cancer EJ-1 cells.</a> Chem. Res. Toxicol. 24, 1586-1596 (2011).</li><li>Wallschlager, D., London, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+methylated+arsenic-sulfur+compounds+in+groundwater.">Determination of methylated arsenic-sulfur compounds in groundwater.</a> Environ. Sci. Technol. 42, 228-234 (2008).</li><li>Zhang, J., Kim, H., Townsend, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methodology+for+assessing+thioarsenic+formation+potential+in+sulfidic+landfill+environments.">Methodology for assessing thioarsenic formation potential in sulfidic landfill environments.</a> Chemosphere. 107, 311-318 (2014).</li><li>Shimoda, Y., 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=Proposal+for+novel+metabolic+pathway+of+highly+toxic+dimethylated+arsenics+accompanied+by+enzymatic+sulfuration,+desulfuration+and+oxidation.">Proposal for novel metabolic pathway of highly toxic dimethylated arsenics accompanied by enzymatic sulfuration, desulfuration and oxidation.</a> Trace Elem. Med. Biol. 30, 129-136 (2015).</li><li>Naranmandura, H., Suzuki, T. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formation+of+dimethylthioarsenicals+in+red+blood+cells.">Formation of dimethylthioarsenicals in red blood cells.</a> Toxicol. Appl. Pharmacol. 227, 390-399 (2008).</li><li>Leffers, L., Ebert, F., Taleshi, S. M., Francesconi, A. K., Schwerdtle, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+toxicological+characterization+of+two+arsenosugars+and+their+metabolites.">In vitro toxicological characterization of two arsenosugars and their metabolites.</a> Mol. Nutr. Food Res. 57, 1270-1282 (2013).</li><li>Wang, Q. Q., Thomas, J. D., Naranmandura, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Important+of+being+thiomethylated:+Formation,+Fate+and+Effects+of+methylated+thioarsenicals.">Important of being thiomethylated: Formation, Fate and Effects of methylated thioarsenicals.</a> Chem. Res. Toxicol. 25, 281-289 (2015).</li><li>Kim, Y. T., Lee, H., Yoon, H. O., Woo, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetics+of+dimethylated+thioarsenicals+and+the+formation+of+highly+toxic+dimethylmonothioarsinic+acid+in+environment.">Kinetics of dimethylated thioarsenicals and the formation of highly toxic dimethylmonothioarsinic acid in environment.</a> Environ. Sci. Technol. 50, 11637-11645 (2016).</li><li>Cullen, W. 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=Methylated+and+thiolated+arsenic+species+for+environmental+and+health+research+-+A+review+on+synthesis+and+characterization.">Methylated and thiolated arsenic species for environmental and health research - A review on synthesis and characterization.</a> J. Environ. Sci. 49, 7-27 (2016).</li><li>Fricke, M., 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=Chromatographic+separation+and+identification+of+products+form+the+reaction+of+dimethylarsinic+acid+with+hydrogen+sulfide.">Chromatographic separation and identification of products form the reaction of dimethylarsinic acid with hydrogen sulfide.</a> Chem. Res. Toxicol. 18, 1821-1829 (2005).</li><li>Fricke, M., Zeller, M., Cullen, W., Witkowski, M., Creed, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dimethylthioarsinic+anhydride:+a+standard+for+arsenic+speciation.">Dimethylthioarsinic anhydride: a standard for arsenic speciation.</a> Anal. Chim. Acta. 583, 78-83 (2007).</li><li>Suzuki, K. T., Iwata, K., Naranmandura, H., Suzuki, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolic+differences+between+twon+dimethylthioarsenicals+in+rats.">Metabolic differences between twon dimethylthioarsenicals in rats.</a> Toxicol. Appl. Pharmacol. 218, 166-173 (2007).</li><li>Jeong, 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=Development+of+a+simultaneous+analytical+method+to+determine+arsenic+speciation+using+HPLC-ICP-MS:+Arsenate,+arsenite,+monomethylarsonic+acid,+dimethylarsinic+acid,+dimethyldithioarsinic+acid,+and+dimethylmonothioarsinic+acid.">Development of a simultaneous analytical method to determine arsenic speciation using HPLC-ICP-MS: Arsenate, arsenite, monomethylarsonic acid, dimethylarsinic acid, dimethyldithioarsinic acid, and dimethylmonothioarsinic acid.</a> Microchem. J. 134, 295-300 (2017).</li><li>Li, Y., Low, C. -. K., Scott, A. J., Amal, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arsenic+speciation+in+municipal+landfill+leachate.">Arsenic speciation in municipal landfill leachate.</a> Chemosphere. 79, 794-801 (2010).</li><li>Conklin, D. S., Fricke, W. M., Creed, A. P., Creed, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+of+the+pH+effects+on+the+formation+of+methylated+thio-arsenicals,+and+the+effects+of+pH+and+temperature+on+their+stability.">Investigation of the pH effects on the formation of methylated thio-arsenicals, and the effects of pH and temperature on their stability.</a> J. Anal. At. Spectrom. 23, 711-716 (2008).</li><li>Hansen, H. R., Raab, A., Jaspara, M., Milne, F. B., Feldmann, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sulfur-containing+arsenical+mistaken+for+dimethylarsinous+acid+[DMA(III)]+and+identified+as+a+natural+metabolite+in+urine:+major+implications+for+studies+on+arsenic+metabolism+and+toxicity.">Sulfur-containing arsenical mistaken for dimethylarsinous acid [DMA(III)] and identified as a natural metabolite in urine: major implications for studies on arsenic metabolism and toxicity.</a> Chem. Res. Toxicol. 17, 1086-1091 (2004).</li><li>Mandal, B. K., Suzuki, K. T., Anzai, K., Yamaguchi, K., Sei, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+SEC-HPLC-ICP-MS+hyphenated+technique+for+identification+of+sulfur-containing+arsenic+metabolites+in+biological+samples.">A SEC-HPLC-ICP-MS hyphenated technique for identification of sulfur-containing arsenic metabolites in biological samples.</a> J. Chromatogr. B. 874, 64-76 (2008).</li><li>Bartel, M., Ebert, F., Leffers, L., Karst, U., Schwerdtle, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toxicological+characterization+of+the+inorganic+and+organic+arsenic+metabolite+thio-DMAV+in+cultured+human+lung+cells.">Toxicological characterization of the inorganic and organic arsenic metabolite thio-DMAV in cultured human lung cells.</a> J. Toxicol. 2011, (2011).</li><li>An, J., 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=Formation+of+dimethyldithioarsinic+acid+in+a+simulated+landfill+leachate+in+relation+to+hydrosulfide+concentration.">Formation of dimethyldithioarsinic acid in a simulated landfill leachate in relation to hydrosulfide concentration.</a> Environ. Geochem. Health. 38, 255-263 (2016).</li><li>Chen, B., 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=Arsenic+speciation+in+the+blood+of+arsenite-treated+F344+rats.">Arsenic speciation in the blood of arsenite-treated F344 rats.</a> Chem. Res. Toxicol. 26, 952-962 (2013).</li><li>Alava, P., 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=HPLC-ICP-MS+method+development+to+monitor+arsenic+speciation+changes+by+human+gut+microbiota.">HPLC-ICP-MS method development to monitor arsenic speciation changes by human gut microbiota.</a> Biomed. Chromatogr. 26, 524-533 (2012).</li><li>Kurosawa, 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=A+novel+metabolic+activation+associated+with+glutathione+in+dimethylmonoarsinic+acid+(DMMTAV)-induced+toxicity+obtained+from+in+vitro+reaction+of+DMMTAV+with+glutathione.">A novel metabolic activation associated with glutathione in dimethylmonoarsinic acid (DMMTAV)-induced toxicity obtained from in vitro reaction of DMMTAV with glutathione.</a> J. trace Elem. Med. Biol. 33, 87-94 (2016).</li></ol>

著者が明らかに何もありません。

開発されたプロトコルが重要なステップ、前研究1,3,4,8,15省略または省略する難しさまたは中の障害になった可能性を明らかにDMMTAV DMDTAVの合成。DMMTAVは酸化に敏感な1、5その合成のための化学試薬調製 N...

ジメチルアルシン酸 (DMAV)、急性毒性とメチル化と摂取1,2時なチオール化による遺伝毒性の両方を展示する実証されているなチオール化ヒ素の代謝経路は集中的にされて学びの in vitroとin vivoの両方3,4同様環境メディア (例えば、埋め立て地の浸出水)5,6のように。以前の研究で発見した両方が減少し、リビングの DMAVのチオール類縁体細胞、dimethyldithioarsinic 酸 (dimethylmonothioarsinic 酸 (DMMTAV)、例, dimethylarsinous 酸 (DMAIII)DMDTAV)7,8,9, 欠損 thioarsenicals DMMTAV他の無機または有機ヒ素10知られているより大きい毒性の展示など。猛毒の thioarsenicals の豊富さは深刻な環境影響を以来、彼らは人間および高い硫化水素の条件11の下で環境に危険をもたらす可能性があります。ただし、DMMTAV DMDTAV (トランス) 形成と環境中運命のメカニズムもさらなる研究が必要です。したがって、thioarsenicals の定量分析は、DMMTAV DMDTAVの環境への影響の理解を改善するために必要です。
標準的な化学物質は、定量分析の重要な条件です、DMMTAV DMDTAVの基準が他の種に種変換の高危険のための先行研究を複製することによって得ることが困難と要員の合成手順12。さらに、参照されるメソッドには、標準化学物質を合成し、定量分析を実行する実用的な困難につながる可能性があります制限があります。DMMTAV DMDTAV一般的で一定モル比1またはバブリング H2S4ガス DMAV 13, 14のソリューションを通じて、DMAVナ2S、H2を混合して用意しています。酸素の毒性が高く経験の浅いユーザーの制御が困難な H2S ガスの直接供給を使用して硫黄のバブリング法機能置換。逆に、上記混合法1DMMTAV DMDTAV環境研究5,6,12の定性分析に広く使用機能のなチオール化H2S のため直接と比較して4 DMMTAV DMDTAV、対象化学物質を生成する化学量論的制御を簡単に作り出す Na2S と H2を混合することによって生成された DMAVH2S ガスを使用します。
つながる可能性があります彼らの重要な実験手順のいくつかでこの研究展示の制限に記載されている混合法手順1,3,4,8,15参照実験に失敗しました。たとえば、特定溶媒 (すなわち、脱イオン水) 準備と抽出の詳細と合成のヒ素の結晶化が過剰省略または十分な細部で記述されていません。このような分散し、手順についての限られた情報が thioarsenicals と信頼性の定量化分析の一貫性のない形成につながる可能性があります。したがって、ここに開発された変更されたプロトコルは、DMMTAVと DMDTAV定量的種の分離分析と原液の合成について説明します。

<table width="712">
	<tbody>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">Cacodylic acid</td>
			<td style="width:179px;">Sigma-Aldrich</td>
			<td style="width:179px;">20835-10G-F</td>
			<td style="width:179px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Sodium sulfide nonahydrate</td>
			<td style="width:179px;">Sigma-Aldrich</td>
			<td style="width:179px;">S2006-500G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">Sulfuric acid 96%</td>
			<td style="width:179px;">J.T.Baker</td>
			<td style="width:179px;">0000011478</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">Ammonium acetate</td>
			<td style="width:179px;">Sigma-Aldrich</td>
			<td style="width:179px;">A7262-500G</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Formic acid 98%</td>
			<td style="width:179px;">Wako Pure Chemical Industries, Ltd.</td>
			<td style="width:179px;">066-00461</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">Diethyl ether (Extra Pure)</td>
			<td style="width:179px;">Junsei Chemical</td>
			<td style="width:179px;">33475-0380</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Adapter cap for 60 mL Bond Elut catridges</td>
			<td style="width:179px;">Agilent Technologies</td>
			<td style="width:179px;">12131004</td>
			<td>Syringe type of SPE</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">Bond Elut C18 cartridge</td>
			<td style="width:179px;">Agilent Technologies</td>
			<td style="width:179px;">14256031</td>
			<td>Syringe type of SPE</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">HyPURITY C-18</td>
			<td style="width:179px;">Thermo Scientific</td>
			<td style="width:179px;">22105-254630</td>
			<td>5 um, 125 x 4.6 mm</td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Glovebox</td>
			<td style="width:179px;">Chungae-chun, Rep. of Korea</td>
			<td style="width:179px;"></td>
			<td>Customized&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Agilent 1260 Infinity Bio-inert LC</td>
			<td style="width:179px;">Agilent Technologies</td>
			<td style="width:179px;">DEAB600252, DEACH00245</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Agilent Technologies 7700 Series ICP-MS</td>
			<td style="width:179px;">Agilent Technologies</td>
			<td style="width:179px;">JP12031510</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="width:179px;height:63px;">Finnigan LCQ Deca XP MAX Mass Spectrometer System</td>
			<td style="width:179px;">Thermo Electron Corporation</td>
			<td style="width:179px;">LDM10627</td>
			<td></td>
		</tr>
	</tbody>
</table>

preparation of dmmtav and dmdtav using dmav for environmental applications: synthesis, purification, and confirmation

ジメチルアルシン酸 (DMAV) なチオール化の代謝経路によって生成される、dimethylmonothioarsinic 酸 (DMMTAV) などの欠損 thioarsenicals および dimethyldithioarsinic 酸 (DMDTAV) は最近ずっと環境だけでなく、人間の臓器で発見します。欠損 thioarsenicals と環境メディアの安定性の生態学的影響を判断する DMMTAV DMDTAVを定量化することができます。これらの化合物の合成法は標準化され、挑戦的な先行研究を複製することです。さらに、化合物種変換なしのストレージを含むストレージの技術についての情報の欠如があります。また、合成法について限られた情報のみが使用できる標準化学物質を合成し、定量分析を実行するのに実験的な困難がある可能性があります。ここに提示されたプロトコル欠損 thioarsenicals、DMMTAV DMDTAV、実質的に変更された合成方法を提供し、高速液体を使用して種の分離分析の定量化に役立つ誘導結合プラズマ質量分析法 (高速液体クロマトグラフィー-ICP-MS) と組み合わせてクロマトグラフィー。化学試薬、ろ過方法、ストレージの準備に焦点を当てたこの手順の実験の手順が変更されました。

DMMTAVは、誤って DMAIII合成法19によって準備されている、合成 DMMTAV DMDTAVの検証は合成、抽出と理想的な基準を決定するための重要なステップ化学物質。合成化学物質はピークの DMMTAV (MW 154 g·mol-1)、エレクトロ スプレー イオン化質量のいずれかの正または負のイオン モードを使用して DMDTAV (17...

Watch this Scientific Journal Video about Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation at JoVE.com

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

DMMTAV DMDTAV環境用 DMAVを使用しての作製: 合成、精製、および確認

本稿では dimethylmonothioarsinic 酸 (DMMTAV) の実験的プロトコルの修正と dimethyldithioarsinic 酸 (DMDTAV) の合成、DMAV の混合によってジメチルアルシン酸 (DMAV) なチオール化を誘導します。、Na2S と H24。修正されたプロトコルは、それにより定量分析における実験的障害を引き起こしている可能性が合成手順の制限の克服、実験ガイドラインを提供します。

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV), which are produced by the metabolic ...

preparation-dmmtav-dmdtav-using-dmav-for-environmental-applications

Research

JoVE Journal

Environment

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

5.4K ビュー.  Korea Basic Science Institute, Seoul Center. 本稿では dimethylmonothioarsinic 酸 (DMMTAV) の実験的プロトコルの修正と dimethyldithioarsinic 酸 (DMDTAV) の合成、DMAV の混合によってジメチルアルシン酸 (DMAV) なチオール化を誘導します。、Na2S と H24。修正されたプロトコルは、それにより定量分析における実験的障害を引き起こしている可能性が合成手順の...

ビデオ: Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

Waste Water Derived Electroactive Microbial Biofilms: Growth, Maintenance, and Basic Characterization

The growth of anodic electroactive microbial biofilms from waste water inocula in a fed-batch reactor is demonstrated using a three-electrode setup controlled by a potentiostat. Thereby the use of potentiostats allows an exact adjustment of the electrode potential and ensures reproducible microbial culturing conditions. During growth the current production is monitored using chronoamperometry (CA). Based on these data the maximum current density (jmax) and the coulombic efficiency (CE) are discussed as measures for characterization of the bioelectrocatalytic performance. Cyclic voltammetry (CV), a nondestructive, i.e. noninvasive, method, is used to study the extracellular electron transfer (EET) of electroactive bacteria. CV measurements are performed on anodic biofilm electrodes in the presence of the microbial substrate, i.e. turnover conditions, and in the absence of the substrate, i.e. nonturnover conditions, using different scan rates. Subsequently, data analysis is exemplified and fundamental thermodynamic parameters of the microbial EET are derived and explained: peak potential (Ep), peak current density (jp), formal potential (Ef) and peak separation (&#916;Ep). Additionally the limits of the method and the state-of the art data analysis are addressed. Thereby this video-article shall provide a guide for the basic experimental steps and the fundamental data analysis.

Chronoamperometric growth of anodic electroactive microbial biofilms in a fed-batch reactor using a three-electrode setup, controlled by a potentiostat, is demonstrated. The extracellular electron transfer is characterized using cyclic voltammetry in the presence of the electron donor and in the absence of the electron donor. Fundamental data analysis is demonstrated.

The growth of anodic electroactive microbial biofilms from waste water inocula in a fed-batch reactor is demonstrated using a three-electrode setup ...

環境学

A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is not an appropriate measure for its availability; bioavailability rather depends on the dynamic interplay of potential mass transfer (flux) of a compound to a microbial cell and the capacity of the latter to degrade the compound. In water-unsaturated parts of the soil, mycelia have been shown to overcome bioavailability limitations by actively transporting and mobilizing organic compounds over the range of centimeters. Whereas the extent of mycelia-based transport can be quantified easily by chemical means, verification of the contaminant-bioavailability to bacterial cells requires a biological method. Addressing this constraint, we chose the PAH fluorene (FLU) as a model compound and developed a water unsaturated model microcosm linking a spatially separated FLU point source and the FLU degrading bioreporter bacterium Burkholderia sartisoli RP037-mChe by a mycelial network of Pythium ultimum. Since the bioreporter expresses eGFP in response of the PAH flux to the cell, bacterial FLU exposure and degradation could be monitored directly in the microcosms via confocal laser scanning microscopy (CLSM). CLSM and image analyses revealed a significant increase of the eGFP expression in the presence of P. ultimum compared to controls without mycelia or FLU thus indicating FLU bioavailability to bacteria after mycelia-mediated transport. CLSM results were supported by chemical analyses in identical microcosms. The developed microcosm proved suitable to investigate contaminant bioavailability and to concomitantly visualize the involved bacteria-mycelial interactions.

A whole cell bioreporter assay with Burkholderia sartisoli RP037-mChe was developed to detect fractions of an organic contaminant (i.e., fluorene) available for bacterial degradation after active transport by mycelia bridging air-filled pores in a water unsaturated model system.

水不飽和システムにおける有機汚染物質の輸送および生物学的利用能を評価するために、全細胞バイオレポーターアプローチ

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is ...

Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity sensor. A monolayer of RTgill-W1 cells forms on the sensing electrodes enclosed within the biochips. The biochips are then used for testing in a field portable electric cell-substrate impedance sensing (ECIS) device designed for rapid toxicity testing of drinking water. The manuscript further describes how to run a toxicity test using the prepared biochips. A control water sample and the test water sample are mixed with pre-measured powdered media and injected into separate channels of the biochip. Impedance readings from the sensing electrodes in each of the biochip channels are measured and compared by an automated statistical software program. The screen on the ECIS instrument will indicate either "Contamination Detected" or "No Contamination Detected" within an hour of sample injection. Advantages are ease of use and rapid response to a broad spectrum of inorganic and organic chemicals at concentrations that are relevant to human health concerns, as well as the long-term stability of stored biochips in a ready state for testing. Limitations are the requirement for cold storage of the biochips and limited sensitivity to cholinesterase-inhibiting pesticides. Applications for this toxicity detector are for rapid field-portable testing of drinking water supplies by Army Preventative Medicine personnel or for use at municipal water treatment facilities.

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial cells for use in a field portable electric cell-substrate impedance sensor. The protocol for running a rapid drinking water toxicity test with the sensor is also described.

毒性のための飲料水試料の迅速な評価のためのRTgill-W1細胞とのインピーダンスベースの流体バイオチップの製造および試験

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity ...

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in nutrient and gas cycles. After cores with intact sediment-water interfaces are collected, they are submerged in incubation tanks and kept under aerobic conditions at in situ temperatures. To initiate a time course of overlying water chemistry, cores are sealed without bubbles using a top cap with a suspended stirrer. Time courses of 4-7 sample points are used to determine the rate of sediment water exchange. Artificial illumination simulates day-time conditions for shallow photosynthetic sediments, and in conjunction with dark incubations can provide net exchanges on a daily basis. The net measurement of N2 is made possible by sampling a time course of dissolved gas concentrations, with high precision mass spectrometric analysis of N2:Ar ratios providing a means to measure N2 concentrations. We have successfully applied this approach to lakes, reservoirs, estuaries, wetlands and storm water ponds, and with care, this approach provides valuable information on biogeochemical balances in aquatic ecosystems.

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

Here, we present small core incubations for the measurement of sediment-water gas and solute exchange. These will provide reliable measurements of sediment-water exchange that assess the role of sediment in influencing biological and biogeochemical processes in aquatic ecosystems.

Oの底生交流<sub&gt; 2</sub&gt;、N<sub&gt; 2</sub小さなコアインキュベーションを使用&gt;と溶存栄養素

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in ...

Use of a Filter Cartridge for Filtration of Water Samples and Extraction of Environmental DNA

Recent studies demonstrated the use of environmental DNA (eDNA) from fishes to be appropriate as a non-invasive monitoring tool. Most of these studies employed disk fiber filters to collect eDNA from water samples, although a number of microbial studies in aquatic environments have employed filter cartridges, because the cartridge has the advantage of accommodating large water volumes and of overall ease of use. Here we provide a protocol for filtration of water samples using the filter cartridge and extraction of eDNA from the filter without having to cut open the housing. The main portions of this protocol consists of 1) filtration of water samples (water volumes &#8804;4 L or &gt;4 L); (2) extraction of DNA on the filter using a roller shaker placed in a preheated incubator; and (3) purification of DNA using a commercial kit. With the use of this and previously-used protocols, we perform metabarcoding analysis of eDNA taken from a huge aquarium tank (7,500 m3) with known species composition, and show the number of detected species per library from the two protocols as the representative results. This protocol has been developed for metabarcoding eDNA from fishes, but is also applicable to eDNA from other organisms.

We describe a protocol for filtration of water samples with a filter cartridge and extraction of environmental DNA (eDNA) without having to cut open the housing to remove the filter. This protocol is developed for metabarcoding eDNA from fishes, but is also applicable to eDNA from other organisms.

ろ過水のサンプルおよび環境DNAの抽出のためのフィルターカートリッジの使用

Recent studies demonstrated the use of environmental DNA (eDNA) from fishes to be appropriate as a non-invasive monitoring tool. Most of these studies ...

Resurrection of Dormant Daphnia magna: Protocol and Applications

Long-term studies enable the identification of eco-evolutionary processes that occur over extended time periods. In addition, they provide key empirical data that may be used in predictive modelling to forecast evolutionary responses of natural ecosystems to future environmental changes. However, excluding a few exceptional cases, long-term studies are scarce because of logistic difficulties associated with accessing temporal samples. Temporal dynamics are frequently studied in the laboratory or in controlled mesocosm experiments with exceptional studies that reconstruct the evolution of natural populations in the wild.
Here, a standard operating procedure (SOP) is provided to revive or resurrect dormant Daphnia magna, a widespread zooplankton keystone species in aquatic ecosystems, to dramatically advance the state-of-the-art longitudinal data collection in natural systems. The field of Resurrection Ecology was defined in 1999 by Kerfoot and co-workers, even though the first attempts at hatching diapausing zooplankton eggs date back to the late 1980s. Since Kerfoot&#39;s seminal paper, the methodology of resurrecting zooplankton species has been increasingly frequently applied, though propagated among laboratories only via direct knowledge transfer. Here, an SOP is described that provides a step-by-step protocol on the practice of resurrecting dormant Daphnia magna eggs.
Two key studies are provided in which the fitness response of resurrected Daphnia magna populations to warming is measured, capitalizing on the ability to study historical and modern populations in the same settings. Finally, the application of next generation sequencing technologies to revived or still dormant stages is discussed. These technologies provide unprecedented power in dissecting the processes and mechanisms of evolution if applied to populations that have experienced changes in selection pressure over time.

Resurrection of Dormant Daphnia magna: Protocol and Applications

Long-term studies are essential to understanding the process of evolution and the mechanisms of adaptation. Generally, these studies require commitments beyond the life-time of researchers. Here, a powerful method is described that dramatically advances state-of-the-art data collection to generate longitudinal data in natural systems.

休眠復活オオミジンコ: プロトコルおよびアプリケーション

Long-term studies enable the identification of eco-evolutionary processes that occur over extended time periods. In addition, they provide key empirical ...

Composition and Distribution Analysis of Bioaerosols Under Different Environmental Conditions

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, we described a protocol for multiple analyses of the biological compositions in environmental PM. Five experiments are presented: (1) PM number monitoring by using a laser particle counter; (2) PM collection by using a cyclonic aerosol sampler; (3) PM collection by using a high-volume air sampler with filters; (4) culturable microbes collection by the Andersen six-stage sampler; and (5) detection of biological composition of environmental PM by bacterial 16SrDNA and fungal ITS region sequencing. We selected hazy days and a livestock farm as two typical examples of the application in this protocol. In this study, these two sampling methods, cyclonic aerosol sampler and filter sampler, showed different sampling efficiency. The cyclonic aerosol sampler performed much better in terms of collecting bacteria, while these two methods showed the same efficiency in collecting fungi. Filter samplers can work under low temperature conditions while cyclonic aerosol samplers have a sampling limitation for temperature. A solid impacting sampler, such as an Andersen six-stage sampler, can be used to sample bioaerosols directly into the culture medium, which increases the survival rate of culturable microorganisms. However, this method mainly relies on culture while more than 99% of microbes cannot be cultured. DNA extracted from the culturable bacteria collected by the Andersen six-stage sampler and samples collected by cyclonic aerosol sampler and filter sampler were detected by bacterial 16S rDNA and fungal ITS region sequencing.All the methods above may have wide application in many fields of study, such as environmental monitoring and airborne pathogen detection. From these results, we can conclude that these methods can be used under different conditions and may help other researchers further explore the health impacts of environmental bioaerosols.

Understanding the biological composition of environmental particulate matter is important for the study of its significant impacts on human health and disease spread. Here, we used three types of bioaerosol sampling methods and a biological analysis of airborne microbes to better explore airborne microbial communities under different environmental conditions.

組成と異なる環境条件下におけるバイオエアロゾルの分布解析

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, ...

Identifying Per- and Polyfluorinated Chemical Species with a Combined Targeted and Non-Targeted-Screening High-Resolution Mass Spectrometry Workflow

Historical and emerging per- and polyfluoroalkyl substances (PFASs) have garnered significant interest from the public and government agencies from the local to federal levels. The continuing evolution of PFAS chemistries presents a challenge to the environmental monitoring, where ongoing development of targeted methods necessarily lags the discovery of new chemical compounds. There is a need, therefore, to have forward-looking methodologies that can detect emerging and unexpected compounds, monitor these species over time, and resolve details of their chemical structure to enable future work in human health. To this end, non-targeted analysis by high-resolution mass spectrometry offers a broad base detection approach that can be combined with almost any sample preparation scheme and provides significant capabilities for compound identification after detection. Herein, we describe a solid-phase extraction (SPE) based sample concentration method tuned for shorter chain and more hydrophilic PFAS chemistries, such as per fluorinated ether acids and sulfonates, and describe analysis of samples prepared in this fashion in both targeted and non-targeted modes. Targeted methods provide superior quantification when reference standards are available but are intrinsically limited to expected compounds when performing analysis. In contrast, a non-targeted approach can identify the presence of unexpected compounds and provide some information about their chemical structure. Information about chemical features can be used to correlate compounds across sample locations and track abundance and occurrence over time.

Here, we present a protocol for the sequential targeted quantification and non-targeted analysis of fluorinated compounds in water by mass spectrometry. This methodology provides quantitative levels of known fluorochemical compounds and identifies unknown chemicals in related samples with semi-quantitative estimates of their abundance.

あたりを識別する-ポリフルオロ化学種の結合対象・非対象と検診高分解能質量分析ワークフロー

Historical and emerging per- and polyfluoroalkyl substances (PFASs) have garnered significant interest from the public and government agencies from the ...

Using Caenorhabditis elegans for Studying Trans- and Multi-Generational Effects of Toxicants

Information about toxicities of chemicals are essential in their application and waste management. For chemicals at low concentrations, the long-term effects are very important in judging their consequences in the environment and on human health. In demonstrating long-term influences, effects of chemicals over generations in recent studies provide new insight. Here, we describe protocols for studying effects of chemicals over multiple generations using free-living nematode Caenorhabditis elegans. Two aspects are presented: (1) trans-generational (TG) and (2) multi-generational effect studies, the latter of which is separated to multi-generational exposure (MGE) and multi-generational residual (MGR) effect studies. The TG effect study is robust with a simple purpose to determine whether chemical exposure to parents can result in any residual consequences on offspring. After the effects are measured on parents, sodium hypochlorite solutions are used to kill the parents and keep the offspring so as to facilitate effect measurement on the offspring. The TG effect study is used to determine whether the offspring are affected when their parent is exposed to the pollutants. The MGE and MGR effect study is systematical used to determine whether continuous generational exposure can result in adaptive responses in offspring over generations. Careful pick-up and transfer are used to distinguish generations to facilitate effect measurement on each generation. We also combined protocols to measure locomotion behavior, reproduction, lifespan, biochemical and gene expression changes. Some example experiments are also presented to illustrate the trans- and multi-generational effect studies.

Using Caenorhabditis elegans for Studying Trans- and Multi-Generational Effects of Toxicants

Trans- and multi-generational effects of persistent chemicals are essential in judging their long-term consequences in the environment and on the human health. We provide novel detailed methods for studying trans- and multi-generational effects using free-living nematode Caenorhabditis elegans.

毒性物質のトランスおよび多世代効果の研究のためのカエナルハブ炎エレガンスの使用

Information about toxicities of chemicals are essential in their application and waste management. For chemicals at low concentrations, the long-term ...

Modeling the Size Spectrum for Macroinvertebrates and Fishes in Stream Ecosystems

The size spectrum is an inverse, allometric scaling relationship between average body mass (M) and the density (D) of individuals within an ecological community or food web. Importantly, the size spectrum assumes that individual size, rather than species&#8217; behavioral or life history characteristics, is the primary determinant of abundance within an ecosystem. Thus, unlike traditional allometric relationships that focus on species-level data (e.g., mean species&#8217; body size vs. population density), size spectra analyses are &#8216;ataxic&#8217; &#8211; individual specimens are identified only by their size, without consideration of taxonomic identity. Size spectra models are efficient representations of traditional, complex food webs and can be used in descriptive as well as predictive contexts (e.g., predicting responses of large consumers to changes in basal resources). Empirical studies from diverse aquatic ecosystems have also reported moderate to high levels of similarity in size spectra slopes, suggesting that common processes may regulate the abundances of small and large organisms in very different settings. This is a protocol to model the community-level size spectrum in wadable streams. The protocol consists of three main steps. First, collect quantitative benthic fish and invertebrate samples that can be used to estimate local densities. Second, standardize the fish and invertebrate data by converting all individuals to ataxic units (i.e., individuals identified by size, irrespective of taxonomic identity), and summing individuals within log2 size bins. Third, use linear regression to model the relationship between ataxic M and D estimates. Detailed instructions are provided herein to complete each of these steps, including custom software to facilitate D estimation and size spectra modeling.

This is a protocol to model the size spectrum (scaling relationship between individual mass and population density) for combined fish and invertebrate data from wadable streams and rivers. Methods include: field techniques to collect quantitative fish and invertebrate samples; lab methods to standardize the field data; and statistical data analysis.

河川生態系におけるマクロ無脊椎動物と魚類のサイズスペクトルのモデリング

The size spectrum is an inverse, allometric scaling relationship between average body mass (M) and the density (D) of individuals within an ecological ...