Name: the use of chemostats in microbial systems biology
Uploaded: 2013-10-14T20:45:00
Description: Watch this Scientific Journal Video about The Use of Chemostats in Microbial Systems Biology at JoVE.com

この作品は、ニューヨーク大学のフォームの起動の資金によってサポートされていました。我々は当初、ケモスタットとしてSixforsバイオリアクターの使用を開発し弥勒ダナムとマットブラウアーに感謝します。

ケモスタットを用いて連続培養の原理は、様々な実装を実現することができる。すべてのケモスタットでは、すべてのコンポーネント、2）十分に混合培養、培養容器の3）適切な通気、4）メディアのほか、文化除去の信頼できる手段の無菌性を維持するための1）の方法を持っていることが不可欠である。ここでは、簡単に代替のセットアップに適合させることができる方法を使用して、ケモ...

<ol>
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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth-limiting+intracellular+metabolites+in+yeast+growing+under+diverse+nutrient+limitations.">Growth-limiting intracellular metabolites in yeast growing under diverse nutrient limitations.</a> Mol. Biol. Cell. 21, 198-211 (2010).</li><li>Boer, V. M., de Winde, J. H., Pronk, J. T., Piper, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genome-wide+transcriptional+responses+of+Saccharomyces+cerevisiae+grown+on+glucose+in+aerobic+chemostat+cultures+limited+for+carbon,+nitrogen,+phosphorus,+or+sulfur.">The genome-wide transcriptional responses of Saccharomyces cerevisiae grown on glucose in aerobic chemostat cultures limited for carbon, nitrogen, phosphorus, or sulfur.</a> J. Biol. Chem. 278, 3265-3274 (2003).</li><li>Brauer, M. 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=Coordination+of+growth+rate,+cell+cycle,+stress+response,+and+metabolic+activity+in+yeast.">Coordination of growth rate, cell cycle, stress response, and metabolic activity in yeast.</a> Mol. Biol. Cell. 19, 352-367 (2008).</li><li>Gresham, D., 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=Adaptation+to+diverse+nitrogen-limited+environments+by+deletion+or+extrachromosomal+element+formation+of+the+GAP1+locus.">Adaptation to diverse nitrogen-limited environments by deletion or extrachromosomal element formation of the GAP1 locus.</a> Proc. Natl. Acad. Sci. 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ケモスタットは、成長制御の定常状態での微生物の培養を可能にする。細胞は不変外部環境になり、一定の速度で継続的に成長する。これは、外部環境が連続的に変化しており、細胞増殖速度は、環境と遺伝子型の複雑な相互作用により決定されたバッチ培養法とは対照的である。したがって、バッチ培養上ケモスタット中の微生物を培養するの主な利点は、実験的に、細胞の増殖速度を制?...

細胞の増殖は、遺伝因子と環境因子の相互作用の1,2複雑なネットワークによって調節される。細胞増殖の調節多因子は、その研究にシステムレベルのアプローチを必要とする。しかし、調節された細胞成長の厳密な研究は、実験的に細胞が成長する速度を制御することの困難性により攻撃される。また、最も単純な実験で、細胞外の条件は、彼らが増殖するように細胞が継続的にその環境を変えるように、頻繁にダイナミックで複雑です。定義された、不変かつ制御された環境における細胞増殖速度の実験的な制御を可能にする細胞を培養する方法であって、これらの問題に対する解決策を、ケモスタットによって提供される。 ケモスタットを用いて連続培養する方法は、独立して1950年にモノー3とNovick＆シラード4で記述されていた。もともと構想として、細胞をCONのメディアの一定体積で栽培されているtinually新しいメディアと古い培地および細胞の同時除去（ 図1）の添加により希釈した。結合した常微分方程式（ 図2）のセル密度（x）およびケモスタット容器中の増殖制限栄養素（単数または複数 ）の濃度の変化率を記述する。重要なことに、この連立方程式は、定常状態で、細胞（ すなわち 、指数増殖速度定数）の特異的増殖速度は、速度に等しいことを顕著含意を有する単一の（非ゼロの）安定した定常状態（ 図3）を予測れる培養物（D）希釈する。希釈率を変えることにより、異なる成長速度でかつ栄養制限の異なる条件下での細胞の定常状態の集団を確立することが可能である。 ケモスタットを使用して、成長率の実験的制御がどのように細胞生理学の変化の理解の発展に重要であった成長5,6のレートで。しかし、微生物学的方法のかつての主力は、20世紀後半の間の分子生物学研究の爆発の際にますますあいまいになりました。今日では、微生物および多細胞生物の両方で増殖制御に関心をリニューアルし、システムレベルの解析のためのゲノム規模法の出現は、ケモスタットを使用するためのモチベーションをリニューアルしました。ここでは、細胞増殖速度の正確な制御とケモスタットを使用して一意に可能な外部環境を生かす3つのアプリケーションについて説明します。このような転写産物および代謝産物など - - 協調的な成長率で規制されている第一に、我々は、生体分子の何千もの豊富さは、どのように調査するケモスタットの使用を記載している。第二に、我々はケモスタットは、競合実験を用いて、栄養素が制約された環境では、異なる遺伝子型の間の成長率の差の正確な推定値を得るために使用することができる方法を記載している。第三に、我々はどのようにケモスタットができる記述一定の貧栄養環境下で増殖する細胞の適応進化を研究するために使用される。これらの例は、ケモスタット環境相互作用および適応進化による細胞増殖調節遺伝子のシステム·レベルの調査を可能にされる方法を例示する。

<table border="1">
	<tbody>
		<tr>
			<td>Name of the reagent</td>
			<td>Company</td>
			<td>Catalogue number</td>
			<td>Comments (optional)</td>
		</tr>
		<tr>
			<td>Infors-HT Sixfors Chemostat</td>
			<td>Appropriate Technical Resources, Inc.</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Glass Bottle 9.5 L</td>
			<td>Fisher Scientific</td>
			<td>02-887-1</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Pinchcock</td>
			<td>Fisher Scientific</td>
			<td>05-867</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Stopper, Size 12, Green Neoprene</td>
			<td>Cole-Palmer</td>
			<td>EW-62991-42</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Straight Connector</td>
			<td>Cole-Palmer</td>
			<td>EW-30703-02</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>General purpose ties 4 in</td>
			<td>Fisher Scientific</td>
			<td>NC9557052</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Tubing, Silicone Rubber</td>
			<td>Small Parts</td>
			<td>B000FMWTDE</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Tubing, Silicone, 3/8 in OD</td>
			<td>Fisher Scientific</td>
			<td>02-587-1Q</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Tubing, Silicone, 7/32 in OD</td>
			<td>Fisher Scientific</td>
			<td>02-587-1E</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Tubing, Stainless Steel, 3/16 in OD</td>
			<td>McMaster-Carr</td>
			<td>6100K164</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Tubing, Stainless Steel, 3/8 in OD</td>
			<td>McMaster-Carr</td>
			<td>6100K161</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Hook Connectors</td>
			<td>Fisher Scientific</td>
			<td>14-66-18Q</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Ratchet Clamp</td>
			<td>Cole-Palmer</td>
			<td>EW-06403-11</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Luer, Female</td>
			<td>Cole-Palmer</td>
			<td>EW-45512-34</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Luer, Male</td>
			<td>Cole-Palmer</td>
			<td>EW-45513-04</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Millipore Aervent MTGR05010 62 mm Filter, 0.2 &mu;m</td>
			<td>Fisher Scientific</td>
			<td>MTGR05010</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>PTFE Acrodisc CR 13 mm filters, 0.2 &mu;m</td>
			<td>Fisher Scientific</td>
			<td>NC9131037</td>
			<td>For Media Vessel and Hosing</td>
		</tr>
		<tr>
			<td>Direct-Reading Flowtube for Air</td>
			<td>Cole-Palmer</td>
			<td>EW-32047-77</td>
			<td>For Nitrogen Gas Setup</td>
		</tr>
		<tr>
			<td>Direct-Reading Flowtube for Nitrogen</td>
			<td>Cole-Palmer</td>
			<td>EW-32048-63</td>
			<td>For Nitrogen Gas Setup</td>
		</tr>
		<tr>
			<td>Gas Proportioner Multitube Frames</td>
			<td>Cole-Palmer</td>
			<td>EW-03218-50</td>
			<td>For Nitrogen Gas Setup</td>
		</tr>
		<tr>
			<td>Regulator, Two-Stage Analytical</td>
			<td>Airgas</td>
			<td>Y12-N145D580</td>
			<td>For Nitrogen Gas Setup</td>
		</tr>
		<tr>
			<td>Hose Adaptor, Stainless Steel</td>
			<td>Airgas</td>
			<td>Y99-26450</td>
			<td>For Nitrogen Gas Setup</td>
		</tr>
		<tr>
			<td>Hose Male Adaptor</td>
			<td>Airgas</td>
			<td>WES544</td>
			<td>For Nitrogen Gas Setup</td>
		</tr>
		<tr>
			<td>Norprene Tubing</td>
			<td>US Plastics</td>
			<td>57280</td>
			<td>For Nitrogen Gas Setup</td>
		</tr>
		<tr>
			<td>Tripod Base</td>
			<td>Cole-Palmer</td>
			<td>EW-03218-58</td>
			<td>For Nitrogen Gas Setup</td>
		</tr>
		<tr>
			<td>Valve Cartridges</td>
			<td>Cole-Palmer</td>
			<td>EW-03217-92</td>
			<td>For Nitrogen Gas Setup</td>
		</tr>
		<tr>
			<td>Carboy 10 L</td>
			<td>Fisher Scientific</td>
			<td>02-963-2A</td>
			<td>For Media Preperation</td>
		</tr>
		<tr>
			<td>Steritop Sterile Vacuum Bottle-Top Filters, 1,000 ml, PES membrane; for 45 mm neck size</td>
			<td>Fisher Scientific</td>
			<td>SCGP-T10-RE</td>
			<td>For Media Preperation</td>
		</tr>
		<tr>
			<td>Media Bottle 100 ml, 45 mm neck size</td>
			<td>Fisher Scientific</td>
			<td>FB-800-100</td>
			<td>For Media Preperation</td>
		</tr>
		<tr>
			<td>calcium chloride&middot;2H2O</td>
			<td>Fisher Scientific</td>
			<td>C79-500</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>sodium chloride</td>
			<td>Fisher Scientific</td>
			<td>BP358-1</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>magnesium sulfate&middot;7H2O</td>
			<td>Sigma Aldrich</td>
			<td>230391</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>potassium phosphate monobasic</td>
			<td>Fisher Scientific</td>
			<td>AC424205000</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>ammonium sulfate</td>
			<td>Fisher Scientific</td>
			<td>AC423400010</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>potassium chloride</td>
			<td>Sigma Aldrich</td>
			<td>P9541</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>boric acid</td>
			<td>Sigma Aldrich</td>
			<td>B6768</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>copper sulfate&middot;5H2O</td>
			<td>Sigma Aldrich</td>
			<td>209198</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>potassium iodide</td>
			<td>Sigma Aldrich</td>
			<td>60400</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>ferric chloride&middot;6H2O</td>
			<td>Fisher Scientific</td>
			<td>I88-100</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>manganese sulfate&middot;H2O</td>
			<td>Sigma Aldrich</td>
			<td>230391</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>sodium molybdate&middot;2H2O</td>
			<td>Sigma Aldrich</td>
			<td>M7634</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>zinc sulfate&middot;7H2O</td>
			<td>Fisher Scientific</td>
			<td>Z68-500</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>biotin</td>
			<td>Fisher Scientific</td>
			<td>BP232-1</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>calcium pantothenate</td>
			<td>Fisher Scientific</td>
			<td>AC24330-1000</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>folic acid</td>
			<td>Sigma Aldrich</td>
			<td>F7876</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>inositol (aka myo-inositol)</td>
			<td>Fisher Scientific</td>
			<td>AC12226-1000</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>niacin (aka nicotinic acid)</td>
			<td>Sigma Aldrich</td>
			<td>N4126</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>p-aminobenzoic acid</td>
			<td>Fisher Scientific</td>
			<td>AC14621-2500</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>pyridoxine HCl</td>
			<td>Sigma Aldrich</td>
			<td>P9755</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>riboflavin</td>
			<td>Sigma Aldrich</td>
			<td>R4500-25G</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>thiamine HCl</td>
			<td>Fisher Scientific</td>
			<td>BP892-100</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>Leucine</td>
			<td>Sigma Aldrich</td>
			<td>L8000-100G</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>Uracil</td>
			<td>Sigma Aldrich</td>
			<td>U0750</td>
			<td>Media Reagents</td>
		</tr>
		<tr>
			<td>Dextrose</td>
			<td>Fisher Scientific</td>
			<td>DF0155-08-5</td>
			<td>Media Reagents</td>
		</tr>
	</tbody>
</table>

the use of chemostats in microbial systems biology

細胞は、外界からの信号に応答してそれらの成長速度を調節する。細胞の成長に合わせて、多様な細胞プロセス、高分子合成、代謝や細胞分裂周期に最終的には、コミットメントなど、調整する必要があります。細胞増殖の速度を制御する調節ネットワーク - 遺伝子発現および代謝を含む - ケモスタット実験的細胞増殖速度を制御する方法は、系統的にどのように成長速度に影響を与える細胞プロセスを研究する強力な手段を提供する。世代ケモスタット数百のために維持するとき、細胞増殖を制限する環境条件における微生物の適応進化を研究するために使用することができる。我々は、ケモスタット培養の原理を記載し、それらの動作を示し、それらの様々なアプリケーションの例を提供する。 20世紀半ばの発売後の廃用の期間の後、に新たにゲノムスケール手法の収束terest細胞の成長および適応進化の分子基盤の調節に生物学的研究におけるケモスタットを使用することにルネサンスを刺激されている。

ケモスタットの主な利点は、希釈率を変化させることによって実験的に細胞の増殖速度を制御する能力である。出芽酵母、サッカロマイセス·セレビシエ（Saccharomyces cerevisiae）においては、細胞の形態、細胞分裂周期におけるその位相の情報である。 unbudded細胞（ 図5A）の割合を測定することによって決定されるように、より高い成長速度を有する集団は、活発に分裂する細胞の...

Watch this Scientific Journal Video about The Use of Chemostats in Microbial Systems Biology at JoVE.com

The Use of Chemostats in Microbial Systems Biology

細胞増殖速度が調節されたプロセスおよび細胞生理学の主要な決定因子である。ケモスタットを用いた連続培養は、細胞増殖およびどのようにそれらのネットワークは細胞増殖を最適化するために進化を制御する分子ネットワークの研究を促進する栄養制限することにより、細胞増殖速度の外因性の制御を可能にする。

微生物システム生物学のケモスタットの使用

Cells regulate their rate of growth in response to signals from the external world. As the cell grows, diverse cellular processes must be coordinated ...

the-use-of-chemostats-in-microbial-systems-biology

Research

JoVE Journal

Biology

Investigating the Microbial Community in the Termite Hindgut - Interview

Jared Leadbetter explains why the termite-gut microbial community is an excellent system for studying the complex interactions between microbes.     The symbiotic relationship existing between the host insect and lignocellulose-degrading gut microbes is explained, as well as the industrial uses of these microbes for degrading plant biomass and generating biofuels.

Jared Leadbetter explains why the termite-gut microbial community is an excellent system for studying the complex interactions between microbes. The symbiotic relationship existing between the host insect and lignocellulose-degrading gut microbes is explained, as well as the industrial uses of these microbes for degrading plant biomass and generating biofuels.

インタビュー - シロアリ後腸内微生物群集の調査

Jared Leadbetter explains why the termite-gut microbial community is an excellent system for studying the complex interactions between microbes.     The ...

生物学

Microbial Communities in Nature and Laboratory - Interview

自然と実験室での微生物群集 - インタビュー

Biology of Microbial Communities  - Interview

微生物群集の生物学 - インタビュー

Using Micro-Electro-Mechanical Systems (MEMS) to Develop Diagnostic Tools

Using Micro-Electro-Mechanical Systems MEMS to Develop Diagnostic Tools

診断ツールを開発するマイクロエレクト​​ロメカニカルシステム（MEMS）の使用

The use of Biofeedback in Clinical Virtual Reality: The INTREPID Project

Generalized anxiety disorder (GAD) is a psychiatric disorder characterized by a constant and unspecific anxiety that interferes with daily-life activities. Its high prevalence in general population and the severe limitations it causes, point out the necessity to find new efficient strategies to treat it. Together with the cognitive-behavioral treatments, relaxation represents a useful approach for the treatment of GAD, but it has the limitation that it is hard to be learned. The INTREPID project is aimed to implement a new instrument to treat anxiety-related disorders and to test its clinical efficacy in reducing anxiety-related symptoms. The innovation of this approach is the combination of virtual reality and biofeedback, so that the first one is directly modified by the output of the second one. In this way, the patient is made aware of his or her reactions through the modification of some features of the VR environment in real time. Using mental exercises the patient learns to control these physiological parameters and using the feedback provided by the virtual environment is able to gauge his or her success. The supplemental use of portable devices, such as PDA or smart-phones, allows the patient to perform at home, individually and autonomously, the same exercises experienced in therapist's office. The goal is to anchor the learned protocol in a real life context, so enhancing the patients' ability to deal with their symptoms. The expected result is a better and faster learning of relaxation techniques, and thus an increased effectiveness of the treatment if compared with traditional clinical protocols.

The Project on Virtual Reality Intelligent Multi-sensor System for the Treatment of Anxiety-related Disorders (INTREPID) is aimed at developing a multi-sensor context-aware virtual reality system for the treatment of anxiety-related disorders.

臨床バーチャルリアリティにおけるバイオフィードバックの利用：INTREPIDプロジェクト

Generalized anxiety disorder (GAD) is a psychiatric disorder characterized by a constant and unspecific anxiety that interferes with daily-life ...

Window on a Microworld: Simple Microfluidic Systems for Studying Microbial Transport in Porous Media

Microbial growth and transport in porous media have important implications for the quality of groundwater and surface water, the recycling of nutrients in the environment, as well as directly for the transmission of pathogens to drinking water supplies. Natural porous media is composed of an intricate physical topology, varied surface chemistries, dynamic gradients of nutrients and electron acceptors, and a patchy distribution of microbes. These features vary substantially over a length scale of microns, making the results of macro-scale investigations of microbial transport difficult to interpret, and the validation of mechanistic models challenging. Here we demonstrate how simple microfluidic devices can be used to visualize microbial interactions with micro-structured habitats, to identify key processes influencing the observed phenomena, and to systematically validate predictive models. Simple, easy-to-use flow cells were constructed out of the transparent, biocompatible and oxygen-permeable material poly(dimethyl siloxane). Standard methods of photolithography were used to make micro-structured masters, and replica molding was used to cast micro-structured flow cells from the masters. The physical design of the flow cell chamber is adaptable to the experimental requirements: microchannels can vary from simple linear connections to complex topologies with feature sizes as small as 2 &mu;m. Our modular EcoChip flow cell array features dozens of identical chambers and flow control by a gravity-driven flow module. We demonstrate that through use of EcoChip devices, physical structures and pressure heads can be held constant or varied systematically while the influence of surface chemistry, fluid properties, or the characteristics of the microbial population is investigated. Through transport experiments using a non-pathogenic, green fluorescent protein-expressing Vibrio bacterial strain, we illustrate the importance of habitat structure, flow conditions, and inoculums size on fundamental transport phenomena, and with real-time particle-scale observations, demonstrate that microfluidics offer a compelling view of a hidden world.

Microfluidic devices can be used to visualize complex natural processes in real time and at the appropriate physical scales. We have developed a simple microfluidic device that mimics key features of natural porous media for studying growth and transport of bacteria in the subsurface.

多孔質媒体中の微生物トランスポート研究のためのシンプルなマイクロ流体システム：マイクロワールドでのウィンドウ

Microbial growth and transport in porous media have important implications for the quality of groundwater and surface water, the recycling of nutrients in ...

Determining the Contribution of the Energy Systems During Exercise

One of the most important aspects of the metabolic demand is the relative contribution of the energy systems to the total energy required for a given physical activity. Although some sports are relatively easy to be reproduced in a laboratory (e.g., running and cycling), a number of sports are much more difficult to be reproduced and studied in controlled situations. This method presents how to assess the differential contribution of the energy systems in sports that are difficult to mimic in controlled laboratory conditions. The concepts shown here can be adapted to virtually any sport.
The following physiologic variables will be needed: rest oxygen consumption, exercise oxygen consumption, post-exercise oxygen consumption, rest plasma lactate concentration and post-exercise plasma peak lactate. To calculate the contribution of the aerobic metabolism, you will need the oxygen consumption at rest and during the exercise. By using the trapezoidal method, calculate the area under the curve of oxygen consumption during exercise, subtracting the area corresponding to the rest oxygen consumption. To calculate the contribution of the alactic anaerobic metabolism, the post-exercise oxygen consumption curve has to be adjusted to a mono or a bi-exponential model (chosen by the one that best fits). Then, use the terms of the fitted equation to calculate anaerobic alactic metabolism, as follows: ATP-CP metabolism = A1 (mL . s-1) x t1 (s). Finally, to calculate the contribution of the lactic anaerobic system, multiply peak plasma lactate by 3 and by the athlete&#x2019;s body mass (the result in mL is then converted to L and into kJ).
The method can be used for both continuous and intermittent exercise. This is a very interesting approach as it can be adapted to exercises and sports that are difficult to be mimicked in controlled environments. Also, this is the only available method capable of distinguishing the contribution of three different energy systems. Thus, the method allows the study of sports with great similarity to real situations, providing desirable ecological validity to the study.

This protocol allows researchers focused on exercise and sports sciences to determine the relative contribution of three different energy systems to the total energy expenditure during a large variety of exercises.

運動時のエネルギーシステムの貢献を決定する

One of the most important aspects of the metabolic demand is the relative contribution of the energy systems to the total energy required for a given ...

Linking Predation Risk, Herbivore Physiological Stress and Microbial Decomposition of Plant Litter

The quantity and quality of detritus entering the soil determines the rate of decomposition by microbial communities as well as recycle rates of nitrogen (N) and carbon (C) sequestration1,2. Plant litter comprises the majority of detritus3, and so it is assumed that decomposition is only marginally influenced by biomass inputs from animals such as herbivores and carnivores4,5. However, carnivores may influence microbial decomposition of plant litter via a chain of interactions in which predation risk alters the physiology of their herbivore prey that in turn alters soil microbial functioning when the herbivore carcasses are decomposed6. A physiological stress response by herbivores to the risk of predation can change the C:N elemental composition of herbivore biomass7,8,9 because stress from predation risk increases herbivore basal energy demands that in nutrient-limited systems forces herbivores to shift their consumption from N-rich resources to support growth and reproduction to C-rich carbohydrate resources to support heightened metabolism6. Herbivores have limited ability to store excess nutrients, so stressed herbivores excrete N as they increase carbohydrate-C consumption7. Ultimately, prey stressed by predation risk increase their body C:N ratio7,10, making them poorer quality resources for the soil microbial pool likely due to lower availability of labile N for microbial enzyme production6. Thus, decomposition of carcasses of stressed herbivores has a priming effect on the functioning of microbial communities that decreases subsequent ability to of microbes to decompose plant litter6,10,11.
We present the methodology to evaluate linkages between predation risk and litter decomposition by soil microbes. We describe how to: induce stress in herbivores from predation risk; measure those stress responses, and measure the consequences on microbial decomposition. We use insights from a model grassland ecosystem comprising the hunting spider predator (Pisuarina mira), a dominant grasshopper herbivore (Melanoplus femurrubrum),and a variety of grass and forb plants9.

We present methods to evaluate how predation risk can alter the chemical quality of herbivore prey by inducing dietary changes to meet demands of heightened stress, and how the decomposition of carcasses from these stressed herbivores slows subsequent plant litter decomposition by soil microbes.

捕食リスク、草食動物生理的ストレスと植物リターの微生物分解を結ぶ

The quantity and quality of detritus entering the soil  determines the rate of decomposition by microbial communities as well as  recycle rates of ...

Design and Analysis of Temperature Preference Behavior and its Circadian Rhythm in Drosophila

The circadian clock regulates many aspects of life, including sleep, locomotor activity, and body temperature (BTR) rhythms1,2. We recently identified a novel Drosophila circadian output, called the temperature preference rhythm (TPR), in which the preferred temperature in flies rises during the day and falls during the night 3. Surprisingly, the TPR and locomotor activity are controlled through distinct circadian neurons3. Drosophila locomotor activity is a well&#160;known circadian behavioral output and has provided strong contributions to the discovery of many conserved mammalian circadian clock genes and mechanisms4. Therefore, understanding TPR will lead to the identification of hitherto unknown molecular and cellular circadian mechanisms. Here, we describe how to perform and analyze the TPR assay. This technique not only allows for dissecting the molecular and neural mechanisms of TPR, but also provides new insights into the fundamental mechanisms of the brain functions that integrate different environmental signals and regulate animal behaviors. Furthermore, our recently published data suggest that the fly TPR shares features with the mammalian BTR3. Drosophila are ectotherms, in which the body temperature is typically behaviorally regulated. Therefore, TPR is a strategy used to generate a rhythmic body temperature in these flies5-8. We believe that further exploration of Drosophila TPR will facilitate the characterization of the mechanisms underlying body temperature control in animals.

Design and Analysis of Temperature Preference Behavior and its Circadian Rhythm in Drosophila

We recently identified a novel Drosophila circadian output, temperature preference rhythm (TPR), in which the preferred temperature in flies rises during the day and falls during the night. TPR is regulated independently from another circadian output, locomotor activity. Here&#160;we describe the design and analysis of TPR in Drosophila.

ショウジョウバエにおける温度嗜好挙動とその概日リズムの設計と解析

The circadian clock regulates many aspects of life, including sleep, locomotor activity, and body temperature (BTR) rhythms1,2. We recently identified a ...

Measuring the Stiffness of Ex Vivo Mouse Aortas Using Atomic Force Microscopy

Arterial stiffening is a significant risk factor and biomarker for cardiovascular disease and a hallmark of aging. Atomic force microscopy (AFM) is a versatile analytical tool for characterizing viscoelastic mechanical properties for a variety of materials ranging from hard (plastic, glass, metal, etc.) surfaces to cells on any substrate. It has been widely used to measure the stiffness of cells, but less frequently used to measure the stiffness of aortas. In this paper, we will describe the procedures for using AFM in contact mode to measure the ex vivo elastic modulus of unloaded mouse arteries. We describe our procedure for isolation of mouse aortas, and then provide detailed information for the AFM analysis. This includes step-by-step instructions for alignment of the laser beam, calibration of the spring constant and deflection sensitivity of the AFM probe, and acquisition of force curves. We also provide a detailed protocol for data analysis of the force curves.

Measuring the Stiffness of Ex Vivo Mouse Aortas Using Atomic Force Microscopy

We present detailed protocols for isolation of aortas from mouse and measurement of their elastic modulus using atomic force microscopy.

の剛性を測定します<em&gt; ex vivoで</em原子間力顕微鏡を使用&gt;マウス大動脈

Arterial stiffening is a significant risk factor and biomarker for cardiovascular disease and a hallmark of aging. Atomic force microscopy (AFM) is a ...