현재의 작품은 유럽 연합 인도 표준시 프로그램 (인도 표준시 - 2002-507464 프로젝트 &quot;- - 공포증 &#39;치료를 위해 가상 현실 지능형 멀티 센서 착용할 수있는 시스템 인트 리피드&quot;)에 의해 지원되었다.

일반 불안 장애의 치료에 대한 임상 프로토콜 (갓) 상담 사무실 내부와 외부를 동시에 프로그램입니다. 휴식 프로그램은 풀어과 대조 감각주의하면서 다양한 근육 그룹을 편안하고, &quot;안내 이미지&quot;프로그램을 포함 &quot;프로 그레시브 근육 휴식 교육&quot;(PMRT), 제이콥 슨에 의해 설명 고전적인 절차의 조합을 말합니다. 우리 교육 영상에서는 편안한 가상 환경의 시각적 큐로 대체됩니다. 상담의 사무실 안에 있어요. 환자는 4 주 (주당 2 세션)에 분산 치료 8 교육 세션에 참여할 수 있습니다. 각 세션은 환자의 불안의 척도를 제공하는 임상 평가로 시작하고 중지합니다. 상담자의 사무실에서 세션의 계획 : <ol><li> 환자의 상태의 초기 임상 평가; </li><li> 환자는 그 / 그녀의 생리 매개 변수 (피부 전도성, 심장 속도, 호흡) 기록 biosensors와 연결되어 있습니다. 이러한 매개 변수의 기본 측정 3 분 등록됩니다; </li><li> 환자는 PC와 연결된 헤드 마운트 디스플레이를 착용하고 조종간을 처리; </li><li> 바다에 면한 아름다운 열대 섬 : 환자는 가상 환경을 탐험 시작합니다. 환자는 치료에 의해 기록된 이야기 다음, 배를 타고 섬에 도달하고 그것을 탐험해보세요. 보도에 따라 그 가이드 그 / 그녀의 섬에을 통해 환자가 다른 패널은 다른 대상 지역으로 방향을 나타냅니다 출발점에 도착합니다. 이들 지역의 각 편안한 운동을 제공, 가상 환경의 일부 요소가 직접 자신에 의해 수정 / 그녀의 생리적 매개 변수를 기록하는 동안이 훈련하는 동안, 음성 가이드에 의해 주어진 징후에 따라 환자는 그를 / 자신을 완화하려고 실시간 인치 따라서 환자는 자신의 / 인증 (전통적인 바이오 피드백 기술에서와 같이) 그녀의 수준의 즉각적인 피드백을 받고 있지만, 그 / 그녀가 immersively 탐구되는 가상 환경에 의해 주어진 여분의 가치. 대상 영역에 바인딩된 : <ul><li> 캠프 파이어 : 생리적 매개 변수가 화재의 감소에 대한 환자의 생리 활성 결과의 감소가가 될 때까지 있도록 화재 강도를 제어; </li><li> 파도 : 바다가 완전히 진정 될 때까지 생리 매개 변수가 파도를 제어; </li><li> 폭포 : 물이 아래로 떨어질 멈출 때까지 생리 각성의 감소는 스트림 강도의 감소에 해당합니다. </li><li> 가제보 : 다르게 다른,이 운동이 단어 또는 그 / 그녀의 개인적인 스트레스를 이벤트와 관련된 이미지를 선택할 수 있습니다 환자에 의해 정의됩니다. 이 기술은 스트레스 접종 프로그램으로 제공하고, 환자는 선택한 항목의 크기 변색 - 부동의 감소를 경험할 수 있습니다 - 저 / ​​자신을 완화하는 그녀의 능력 / 자신에 따라; </li></ul></li><li> 일단 가상 현실 세션을 완료, 생리 매개 변수는 휴식 상태에 다시 기록됩니다; </li><li> 환자의 상태의 최종 임상 평가. </li></ol> 바깥 주치의 사무실 : 훈련의 효능을 개선하고 휴식의 효과를 증가하기 위해, 환자 Homecare 시나리오가 발생하여 치료사의 사무실 밖에서 이완 기법을 연습합니다. 이것은 모바일 장치에서 치료 중에 경험 같은 가상 환경을 제시하여 실현됩니다. 모바일 장치는 심장 속도와 피부 전도성을 포함 biosensors 세트에 블루투스를 통해 연결되어 있습니다. 이 장비는 충분히 작고 자율적으로 (그림을 참조하십시오. 1) 환자에 의해 처리되는 사용하기 쉬운 것입니다. 환자는 치료에 의해 제공되는 치료 계획에 따라 치료의 전체 기간 (한 달)에 대한 최소한 하루에 한번 휴식 능력을 훈련해야합니다. 대표 결과 바이오 피드백과 가상 현실을 모두가 휴식을 유도하는 효과적인 것으로 알려져 있기 때문에, 주요 예상 결과는이 두 방법의 조합을 개별적으로 사용되는 두 가지 기술보다 일반적인 불안 장애에 의해 영향을 환자의 불안을 줄이는 데보다 효과적이라는 사실이다. <img src="/files/ftp_upload/1554/Fig1.jpg" alt="그림 1" /> 그림 1 : GSR / HR biosensors 모듈.

<ol>
	<li>Arntz, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cognitive+therapy+versus+applied+relaxation+as+treatment+of+generalized+anxiety+disorder.">Cognitive therapy versus applied relaxation as treatment of generalized anxiety disorder.</a> Behav Res Ther. 41 (6), 633-633 (2003).</li><li>Siev, J., Chambless, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Specificity+of+treatment+effects:+cognitive+therapy+and+relaxation+for+generalized+anxiety+and+panic+disorders.">Specificity of treatment effects: cognitive therapy and relaxation for generalized anxiety and panic disorders.</a> Journal of consulting and clinical psychology. 75 (4), 513-513 (2007).</li><li>Jacobsen, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Progressive relaxation. , (1938).</li><li>Villani, D., Riva, F., Riva, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+technologies+for+relaxation:+the+role+of+presence.">New technologies for relaxation: the role of presence.</a> International journal of Stress Management. 14 (3), 260-260 (2007).</li></ol>

The authors have nothing to disclose.

우리의 가설은 가상 현실과 직접적으로 혼자 가상 현실에 비해, 가상 환경을 수정 바이오 피드백의 조합은보다 빠르고 휴식을 학습의 결과 것입니다. 그렇다면이 새로운 악기가 불안 장애를 치료하기 위해 만든이 적용 심지어 다른 불안 관련 pathologies에서 테스트를 수 있습니다.

the use of biofeedback in clinical virtual reality: the intrepid project

일반 불안 장애 (갓)은 일상 생활 활동을 방해 상수와 unspecific 불안에 의해 특징 정신 장애이다. 는 일반 인구의 높은 유행하고 원인이 심각한 제한은 그것을 치료하는 새로운 효율적인 전략을 찾을 필요성을 지적. 함께인지 - 행동 요법과 함께 휴식을 갓의 치료를 위해 유용한 방법을 나타냅니다 있지만, 그것이 배울 수 어려운 제한이 있습니다. 인트 리피드 프로젝트는 불안과 관련된 장애를 치료하고 불안 관련 증상을 감소 년에 임상 효능을 테스트하는 새로운 계기를 구현하는 것을 목표로하고 있습니다. 이 방법의 혁신은 첫 번째가 바로 두 번째의 출력에 의해 수정되도록, 가상 현실과 바이오 피드백의 조합을 말합니다. 이러한 방법으로, 환자는 실시간으로 VR 환경의 일부 기능의 수정을 통해 자신의 반응 인식 이루어집니다. 환자가 이러한 생리적 매개 변수를 제어하는​​ 배우는 정신 연습을 사용하여 가상 환경에서 제공하는 피드백을 사용하면 자신의 성공 여부를 평가할 수 있습니다. 이러한 PDA 또는 스마트 전화와 같은 휴대용 장치의 보완 사용 환자, 개별적으로 그리고 자율적으로, 가정에서 치료사의 사무실에서 경험 같은 연습을 수행할 수 있습니다. 목표는 그래서 그들의 증상에 대처하기 위해 환자의 능력을 향상, 실제 생활 상황에서 배운 프로토콜을 앵커 것입니다. 예상 결과는 전통적인 임상 프로토콜과 비교하면 따라서 치료의 증가 효과 이완 기술의 학습보다 빠르고이며.

Watch this Scientific Journal Video about The use of Biofeedback in Clinical Virtual Reality: The INTREPID Project at JoVE.com

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

불안과 관련된 장애 (인트 리피드)의 치료를위한 가상 현실 지능형 멀티 센서 시스템에있는 프로젝트는 불안 관련 장애의 치료를위한 멀티 센서 컨텍스트 인식 가상 현실 시스템을 개발 목표로하고 있습니다.

임상 가상 현실의 생체 자기 제어의 사용 인트 리피드 프로젝트

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

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Research

JoVE Journal

Biology

15.0K 조회수.  Istituto Auxologico Italiano. 불안과 관련된 장애 (인트 리피드)의 치료를위한 가상 현실 지능형 멀티 센서 시스템에있는 프로젝트는 불안 관련 장애의 치료를위한 멀티 센서 컨텍스트 인식 가상 현실 시스템을 개발 목표로하고 있습니다.

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

Use of Rotorod as a Method for the Qualitative Analysis of Walking in Rat

High speed videoanalysis of the details of movement can provide a source of information about qualitative aspects of walking movements. When walking on a rotorod, animals remain in approximately the same place making repetitive movements of stepping. Thus the task provides a rich source of information on the details of foot stepping movements. Subjects were hemi-Parkinson analogue rats, produced by injection of 6-hydroxydopamine (6-OHDA) into the right nigrostriatal bundle to deplete nigrostriatal dopamine (DA). The present report provides a video analysis illustration of animals previously were filmed from frontal, lateral, and posterior views as they walked (15). Rating scales and frame-by-frame replay of the video records of stepping behavior indicated that the hemi-Parkinson rats were chronically impaired in posture and limb use contralateral to the DA-depletion. The contralateral limbs participated less in initiating and sustaining propulsion than the ipsilateral limbs. These deficits secondary to unilateral DA-depletion show that the rotorod provides a use task for the analysis of stepping movements.

The rotorod test is used to assess motor status in the walking movement of hemi-Parkinson analogue rats.

쥐에서 도보의 질적 분석 방법으로 Rotorod의 사용

High speed videoanalysis of the details of movement can provide a source of information about qualitative aspects of walking movements. When walking on a ...

연구

생물학

Psychophysiological Stress Assessment Using Biofeedback 

In the last half century, research in biofeedback has shown the extent to which the human mind can influence the functioning of the autonomic nervous system, previously thought to be outside of conscious control. By letting people observe signals from their own bodies, biofeedback enables them to develop greater awareness of their physiological and psychological reactions, such as stress, and to learn to modify these reactions. Biofeedback practitioners can facilitate this process by assessing people s reactions to mildly stressful events and formulating a biofeedback-based treatment plan. During stress assessment the practitioner first records a baseline for physiological readings, and then presents the client with several mild stressors, such as a cognitive, physical and emotional stressor. Variety of stressors is presented in order to determine a person's stimulus-response specificity, or differences in each person's reaction to qualitatively different stimuli. This video will demonstrate the process of psychophysiological stress assessment using biofeedback and present general guidelines for treatment planning.

Psychophysiological Stress Assessment Using Biofeedback

In this video we will describe one method of assessing person's psychophysiological reaction to stress using biofeedback. We will also present general guidelines for treatment planning.

생체 자기 제어를 사용 Psychophysiological 스트레스 평가

In the last half century, research in biofeedback has shown the extent to which the human mind can influence the functioning of the autonomic nervous ...

Use of the Protease Fluorescent Detection Kit to Determine Protease Activity

The Protease Fluorescent Detection Kit provides ready-to-use reagents for detecting the presence of protease activity. This simple assay to detect protease activity uses casein labeled with fluorescein isothiocyanate (FITC) as the substrate.
Protease activity results in the cleavage of the FITC-labeled casein substrate into smaller fragments, which do not precipitate under acidic conditions. After incubation of the protease sample and substrate, the reaction is acidified with the addition of trichloroacetic acid (TCA). The mixture is then centrifuged with the undigested substrate forming a pellet and the smaller, acid soluble fragments remaining in solution. The supernatant is neutralized and the fluorescence of the FITC-labeled fragments is measured.
The described kit procedure detects the trypsin protease control at a concentration of approximately 0.5 &mu;g/ml (5 ng of trypsin added to the assay). This sensitivity can be increased with a longer incubation time, up to 24 hours. The assay is performed in microcentrifuge tubes and procedures are provided for fluorescence detection using either cuvettes or multiwell plates.

The Protease Fluorescent Detection Kit is designed for the measurement of protease activity using fluorometry. It is also suitable for detection of trace amounts of protease contamination. The method is based on the proteolytic hydroysis of a proprietary formulation of a FITC-labeled casein substrate.

단백 분해 효소의 활동을 확인하는 프로 테아제 형광 검출 키트의 사용

The Protease Fluorescent Detection Kit provides ready-to-use reagents for detecting the presence of protease activity. This simple assay to detect ...

The use of SC1 (Pluripotin) to Support mESC Self-renewal in the Absence of LIF

Mouse embryonic stem (ES) cells are conventionally cultured with Leukemia Inhibitory Factor (LIF) to maintain self-renewal.1 However, LIF is expensive and activation of the LIF/JAK/STAT3 pathway is not absolutely required to maintain the self-renewal state.2 The SC1 small molecule may be an economical alternative to LIF. SC1 functions through dual inhibition of Ras-GAP and ERK1.3 Illustration of its mechanism of action makes it a useful tool to study the fundamental molecular mechanism of self-renewal. Here we demonstrate the procedure for culturing mouse ES cells in the presence of SC1 and show that they are able to maintain self-renewal in the absence of LIF. Cells cultured with SC1 showed similar morphology compared to cells maintained with LIF. Both exhibited typical mouse ES morphology after five passages. Expression of typical pluripotency markers (Oct4, Sox2, Nanog, and SSEA1) was observed after five passages in the presence of SC1. Furthermore, SC1 caused no overt toxicity on mouse ES cells.

The use of SC1 Pluripotin to Support mESC Self-renewal in the Absence of LIF

SC1 functions through dual inhibition of Ras- GAP and ERK1. We tested the function of SC1 in supporting mouse ES cell self-renewal in the absence of LIF and showed that SC1 is able to maintain self-renewal of mouse ES cell cultures.

난생의 부재에서 자기 갱신 mESC를 지원하기 위해 SC1 (Pluripotin)의 사용

Mouse embryonic stem (ES) cells are conventionally cultured with Leukemia Inhibitory Factor (LIF) to maintain self-renewal.1 However, LIF is expensive and ...

Purification of the M. magneticum Strain AMB-1 Magnetosome Associated Protein MamA&Delta;41

Magnetotactic bacteria comprise a diverse group of aquatic microorganisms that are able to orientate themselves along geomagnetic fields. This behavior is believed to aid their search for suitable environments (1). This capability is conferred by the magnetosome, a subcellular organelle that consists of a linear-chain assembly of lipid vesicles each able to biomineralize and enclose a ~50-nm crystal of magnetite or greigite. A principle component of the magnetosome that was shown to be required for the formation of functional vesicles is MamA. MamA is a highly abundant magnetosome-associated protein which is one of the most characterized magnetosome-associated proteins in vivo (2-6). This article focuses on the purification of MamA, which despite being studied in vivo, no clear functional or structural details have been identified for it. Bioinformatics analysis suggested that MamA is a tetra-tricopeptide repeat (TPR) containing protein. TPR is a structural motif found as such or forming part of a bigger fold in a wide range of proteins, it serves as a template for protein-protein interactions and mediates multi-protein complexes (7). TPRs are involved in many crucial tasks in eukaryotic cell organelle processes and many bacterial pathways (8-14). In order to understand MamA, a unique TPR containing protein, highly purified protein is required as a first step. In this article, we present the purification protocol for a stable MamA deletion mutant (MamA&Delta;41) from M. magneticum AMB-1.

Purification of the &lt;em&gt;M. magneticum&lt;/em&gt; Strain AMB-1 Magnetosome Associated Protein MamA&amp;Delta;41

MamA is a unique Magnetosome associated protein which was shown to be involved in magnetosome activation. Here we present the purification protocol of MamA deletion mutant (MamA&Delta;41) from M. magneticum AMB-1.

의 정화 M. magneticum 스트레인 AMB - 1 Magnetosome 관련 단백질 MamAΔ41

Magnetotactic bacteria comprise a diverse group of aquatic microorganisms that are able to orientate themselves along geomagnetic fields. This behavior is ...

Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis

The proper elimination of unwanted or aberrant cells through apoptosis and subsequent phagocytosis (apoptotic cell clearance) is crucial for normal development in all metazoan organisms. Apoptotic cell clearance is a highly dynamic process intimately associated with cell death; unengulfed apoptotic cells are barely seen in vivo under normal conditions. In order to understand the different steps of apoptotic cell clearance and to compare 'professional' phagocytes - macrophages and dendritic cells to 'non-professional' - tissue-resident neighboring cells, in vivo live imaging of the process is extremely valuable. Here we describe a protocol for studying apoptotic cell clearance in live Drosophila embryos. To follow the dynamics of different steps in phagocytosis we use specific markers for apoptotic cells and phagocytes. In addition, we can monitor two phagocyte systems in parallel: 'professional' macrophages and 'semi-professional' glia in the developing central nervous system (CNS). The method described here employs the Drosophila embryo as an excellent model for real time studies of apoptotic cell clearance.

Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis

Here we describe an effective method for studying dynamics of apoptotic cell clearance in vivo. This method employs live Drosophila embryos as a powerful model for monitoring phagocytosis of apoptotic cells using specific labeling of apoptotic cells and phagocytes.

사멸 세포 간극의 이미지를하는 동안 라이브<em&gt; 초파리</em&gt; 배아

The  proper elimination of unwanted or aberrant cells through apoptosis and  subsequent phagocytosis (apoptotic cell clearance) is crucial for normal  ...

The Use of Chemostats in Microbial Systems Biology

Cells regulate their rate of growth in response to signals from the external world. As the cell grows, diverse cellular processes must be coordinated including macromolecular synthesis, metabolism and ultimately, commitment to the cell division cycle. The chemostat, a method of experimentally controlling cell growth rate, provides a powerful means of systematically studying how growth rate impacts cellular processes - including gene expression and metabolism - and the regulatory networks that control the rate of cell growth. When maintained for hundreds of generations chemostats can be used to study adaptive evolution of microbes in environmental conditions that limit cell growth. We describe the principle of chemostat cultures, demonstrate their operation and provide examples of their various applications. Following a period of disuse after their introduction in the middle of the twentieth century, the convergence of genome-scale methodologies with a renewed interest in the regulation of cell growth and the molecular basis of adaptive evolution is stimulating a renaissance in the use of chemostats in biological research.

Cell growth rate is a regulated process and a primary determinant of cell physiology. Continuous culturing using chemostats enables extrinsic control of cell growth rate by nutrient limitation facilitating the study of molecular networks that control cell growth and how those networks evolve to optimize cell growth.

미생물 시스템 생물학의 Chemostats의 사용

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

Culturing of Human Nasal Epithelial Cells at the Air Liquid Interface

In vitro models using human primary epithelial cells are essential in understanding key functions of the respiratory epithelium in the context of microbial infections or inhaled agents. Direct comparisons of cells obtained from diseased populations allow us to characterize different phenotypes and dissect the underlying mechanisms mediating changes in epithelial cell function. Culturing epithelial cells from the human tracheobronchial region has been well documented, but is limited by the availability of human lung tissue or invasiveness associated with obtaining the bronchial brushes biopsies. Nasal epithelial cells are obtained through much less invasive superficial nasal scrape biopsies and subjects can be biopsied multiple times with no significant side effects. Additionally, the nose is the entry point to the respiratory system and therefore one of the first sites to be exposed to any kind of air-borne stressor, such as microbial agents, pollutants, or allergens. 
Briefly, nasal epithelial cells obtained from human volunteers are expanded on coated tissue culture plates, and then transferred onto cell culture inserts. Upon reaching confluency, cells continue to be cultured at the air-liquid interface (ALI), for several weeks, which creates more physiologically relevant conditions. The ALI culture condition uses defined media leading to a differentiated epithelium that exhibits morphological and functional characteristics similar to the human nasal epithelium, with both ciliated and mucus producing cells. Tissue culture inserts with differentiated nasal epithelial cells can be manipulated in a variety of ways depending on the research questions (treatment with pharmacological agents, transduction with lentiviral vectors, exposure to gases, or infection with microbial agents) and analyzed for numerous different endpoints ranging from cellular and molecular pathways, functional changes, morphology, etc. 
In vitro models of differentiated human nasal epithelial cells will enable investigators to address novel and important research questions by using organotypic experimental models that largely mimic the nasal epithelium in vivo.

Nasal epithelial cells, obtained through superficial scrape biopsy of human volunteers, are expanded and transferred onto tissue culture inserts. Upon reaching confluency, cells are grown at air liquid interface, yielding cultures of ciliated and non-ciliated cells. Differentiated nasal epithelial cell cultures provide viable experimental models for studying the respiratory mucosa.

에어 리퀴드 인터페이스에서 인간의 비강 상피 세포의 배양

In vitro models using human primary  epithelial cells are essential in understanding key functions of the respiratory  epithelium in the context of ...

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Probe-based quantitative PCR (qPCR) is a favoured method for measuring transcript abundance, since it is one of the most sensitive detection methods that provides an accurate and reproducible analysis. Probe-based chemistry offers the least background fluorescence as compared to other (dye-based) chemistries. Presently, there are several platforms available that use probe-based chemistry to quantitate transcript abundance. qPCR in a 96 well plate is the most routinely used method, however only a maximum of 96 samples or miRNAs can be tested in a single run. This is time-consuming and tedious if a large number of samples/miRNAs are to be analyzed. High-throughput probe-based platforms such as microfluidics (e.g. TaqMan Array Card) and nanofluidics arrays (e.g. OpenArray) offer ease to reproducibly and efficiently detect the abundance of multiple microRNAs in a large number of samples in a short time. Here, we demonstrate the experimental setup and protocol for miRNA quantitation from serum or plasma-EDTA samples, using probe-based chemistry and three different platforms (96 well plate, microfluidics and nanofluidics arrays) offering increasing levels of throughput.

Circulating microRNAs have recently emerged as promising and novel biomarkers for various cancers and other diseases. The goal of this article is to discuss three different probe-based real-time PCR platforms and methods that are available to quantify and determine the abundance of circulating microRNAs.

프로브 기반의 실시간 PCR은 마이크로 RNA의 정량 측정을위한 접근 방법

Probe-based quantitative PCR (qPCR) is a favoured method for measuring transcript abundance, since it is one of the most sensitive detection methods that ...

Use of Enzymatic Biosensors to Quantify Endogenous ATP or H2O2 in the Kidney

Enzymatic microelectrode biosensors have been widely used to measure extracellular signaling in real-time. Most of their use has been limited to brain slices and neuronal cell cultures. Recently, this technology has been applied to the whole organs. Advances in sensor design have made &#160;possible the measuring of&#160;cell signaling in blood-perfused in vivo kidneys. The present protocols list the steps needed to measure ATP and H2O2 signaling in the rat kidney interstitium. Two separate sensor designs are used for the ex vivo and in vivo protocols. Both types of sensor&#160;are coated with&#160;a thin enzymatic biolayer on top of a permselectivity layer to give fast responding, sensitive and selective biosensors. The permselectivity layer protects the signal from the interferents in biological tissue, and the enzymatic layer utilizes&#160;the sequential catalytic reaction of glycerol kinase and glycerol-3-phosphate oxidase in the presence of ATP to produce H2O2. The set of sensors used for the ex vivo studies further detected analyte by oxidation of H2O2 on a platinum/iridium (Pt-Ir) wire electrode. The sensors for the&#160;in vivo studies are instead based on the reduction of H2O2 on a mediator coated gold electrode designed for blood-perfused tissue. Final concentration changes&#160;are detected by real-time amperometry followed by calibration to known concentrations of analyte. Additionally, the specificity of the amperometric signal can be confirmed by the addition of enzymes such as catalase and apyrase that break down H2O2 and ATP correspondingly. These sensors also rely heavily on accurate calibrations before and after each experiment. The following two protocols establish the study of real-time detection of ATP and H2O2 in kidney tissues, and can be further modified to extend the described method for use in other biological preparations or whole organs.

Use of Enzymatic Biosensors to Quantify Endogenous ATP or H2O2 in the Kidney

Enzymatic microelectrode biosensors enable real-time measurements of extracellular cell signaling in biologically-relevant concentrations. The following protocols extend the applications of biosensors to the ex vivo and in vivo detection of ATP and H2O2 in the kidney.

효소 바이오 센서의 사용은 내인성 ATP 또는 H를 정량화하기<sub&gt; 2</sub&gt; O<sub&gt; 2</sub&gt; 신장에

Enzymatic microelectrode biosensors have been widely used to measure extracellular signaling in real-time. Most of their use has been limited to brain ...