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[[File:SUIT-nomenclature.jpg|300px|right|SUIT protocols]]
'''Coupling/pathway control diagrams''' illustrate the respiratory '''states''' obtained step-by-step in substrate-uncoupler-inhibitor titrations in a [[SUIT protocol]]. Each step (to the next state) is defined by an initial state and a [[metabolic control variable]], ''X''. The respiratory states are shown by boxes. ''X'' is usually the titrated substance in a SUIT protocol. If ''X'' ([[ADP]], [[uncoupler]]s, or inhibitors of the [[phosphorylation system]], e.g. oligomycin) exerts '''coupling control''', then a transition is induced between two [[coupling-control state]]s. If ''X'' (fuel substrates, e.g. pyruvate and succinate, or [[Electron transfer pathway]] inhibitors, e.g. rotenone) exerts '''pathway control''', then a transition is induced between two [[Electron-transfer-pathway state]]s. The type of metabolic control (''X'') is shown by arrows linking two respiratory states, with vertical arrows indicating coupling control, and horizontal arrows indicating pathway control. [[Marks - DatLab |Marks]] define the section of an experimental trace in a given [[respiratory state]] (steady state). [[Events - DatLab |Events]] define the titration of ''X'' inducing a transition in the SUIT protocol. The specific sequence of coupling control and pathway control steps defines the [[SUIT protocol pattern]]. The coupling/pathway control diagrams define the [[categories of SUIT protocols]] (see Figure). +
[[Image:Cover-Slip_black.JPG|180px|right]] A '''Cover-Slip''' should be placed on top of the O2k-Stopper to minimize contamination and evaporation of liquid extruding from the capillary of the stopper. The Cover-Slips do not exert any direct effect on oxygen backdiffusion into the [[O2k-chamber]]. Use the the '''Cover-Slip\black''' to avoid light penetration and disturbance of fluorescence signals and generally for optical measurements in the O2k. +
The '''Crabtree effect''' describes the observation that respiration is frequently inhibited when high concentrations of glucose or fructose are added to the culture medium - a phenomenon observed in numerous cell types, particularly in proliferating cells, not only tumor cells but also bacteria and yeast. The Pasteur effect (suppression of glycolysis by oxygen) is the converse of the Crabtree effect (suppression of respiration by high concentration of glucose or fructose). +
'''Creatine''' is a nitrogenous organic acid that occurs naturally in vertebrates and helps primarily muscle cells to supply energy by increasing the formation of adenosine triphosphate ([[ATP]]). +
The mitochondrial '''creatine kinase''', also known as phosphocreatine kinase (CPK), facilitates energy transport with [[creatine]] and [[phosphocreatine]] as diffusible intermediates. +
Open Access preprints (not peer-reviewed) and articles (peer-reviewed) distributed under the terms of the '''Creative Commons Attribution License''' allow unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are credited. © remains with the authors, who have granted the publisher license in perpetuity. +
The '''critical oxygen pressure''', ''p''<sub>c</sub>, is defined as the partial oxygen pressure, ''p''<sub>O2</sub>, below which [[aerobic]] catabolism (respiration or oxygen consumption) declines significantly. If [[anaerobic]] catabolism is activated simultaneously to compensate for lower aerobic ATP generation, then the '''[[limiting oxygen pressure]]''', ''p''<sub>l</sub>, is equal to the ''p''<sub>c</sub>. In many cases, however, the ''p''<sub>l</sub> is substantially lower than the ''p''<sub>c</sub>. +
Coordinated respiratory [[SUIT|SUIT protocols]] are designed to include '''cross-linked respiratory states''', which are common to these protocols. Different SUIT protocols address a variety of respiratory control steps which cannot be accomodated in a single protocol. Cross-linked respiratory states are included in each individual coordinated protocol, such that these states can be considered as replicate measurements, which also allow for harmonization of data obtained with these different protocols. +
'''Curcumin''' has been shown to possess significant anti-inflammatory, anti-oxidant, anti-carcinogenic, anti-mutagenic, anti-coagulant and anti-infective effects. The protective effects of curcumin on rat heart mitochondrial injuries induced by in vitro anoxia–reoxygenation were evaluated by [http://www.ncbi.nlm.nih.gov/pubmed/23984717 Xu et al 2013]. It was found that curcumin added before anoxia or immediately prior to reoxygenation exhibited remarkable protective effects against anoxia–reoxygenation induced oxidative damage to mitochondria. +
A '''Custom label''' can be entered in this box to rename the axis label. Two lines are available for the axis name and unit. +
Stoppers can be custom-made for applications with user-specific sensors according to customer specifications. +
'''Cuvettes''' are used in [[fluorometry]] and [[transmission spectrophotometry]] to contain the samples. Use of the term 'cells' for cuvettes is discouraged, to avoid confusion with 'living cells'. Traditionally cuvettes have a square cross-section (10 x 10 mm). For many applications they are made of transparent plastic. Glass cells are used where samples may contain plastic solvents, and for some applications requiring measurements below 300 nm, quartz glass or high purity fused silica cuvettes may be necessary. +
'''Cyanide''' (usually added as KCN) is a competitive inhibitor of [[Complex_IV| cytochcrome ''c'' oxidase (CIV)]]. Inhibition is reversed by pyruvate and high oxygen levels. +
'''Cyclic voltammetry''' (CV) is a type of electrochemical measurement which is applied with the [[Q-Module]] as quality control to
(''1'') determine the oxidation and reduction peak potentials of [[Coenzyme Q]] in the specific experimental condition, (2) check the quality of the [[Q-Sensor]], and (''3'') test the interference of chemicals used in the HRR assay with the Q-Sensor. In CV, the [[Q-Sensor]] with the [[three-electrode system]] is used to obtain information about the analyte ([[Coenzyme Q|CoQ]]) by measuring the current (''I'') as the electric potential (''V'') between two of the electrodes is varied. In CV the electric potential between the glassy carbon (GC) and the Ag/AgCl reference electrode changes linearly versus time in cyclical phases, while the current is detected between GC and platinum electrode (Pt). The detected current is plotted versus the applied voltage to obtain the typical cyclic voltammogram trace (Figure 1). The presence of substances that are oxidized/reduced will result in current between GC and Pt, which can be seen as characteristic peaks in the voltammogram at a defined potential. The oxidation or the reduction peak potential values are used to set the GC (integrated into the [[Q-Sensor]]) for a separate experiment to measure the [[Q redox state]] of a biological sample. The oxidation and reduction peak potentials can be influenced by 1) the respiration medium, 2) the type of [[Coenzyme Q | CoQ]], 3) the polarization window, 4) the scan speed, 5) the number of cycles, 6) the concentration of the analyte (CoQ), and 7) the initial polarization voltage. <be>
:::-''See'': [[MiPNet24.12 NextGen-O2k: Q-Module]].
:::::[[MiPNet24.16 DatLab8.0: CV-Module]] +
'''Cyclic voltammetry''' +
'''Cyclosporin A''' (CsA) is a cyclic undecapeptide from an extract of soil fungi that binds the cyclophilin D and thus preventing the formation of the mitochondrial [[PTP|permeability transition pore]]. The interaction of CsA with the cyclophilin D is phosphate mediated but the full mechanism of interaction is not well understood. For example, the deficiency of cyclophilin D in KO models does not prevent mitochondria from permeability transition and from CsA inhibition. Moreover, it is also a is a calcineurin inhibitor and potent immunosuppressive agent used largely as a means of prophylaxis against cellular rejection after solid organ transplantation. +
'''Cytochrome ''c''''' is a component of the Electron transfer-pathway ([[Electron transfer pathway]]) in mitochondria. It is a small heme protein loosely associated with the outer side of the inner mitochondrial membrane. The heme group of cytochrome ''c'' transfers electrons from [[Complex III]] to [[Complex IV]]. The release of cytochrome ''c'' into the cytoplasm is associated with apoptosis. Cytochrome ''c'' is applied in [[HRR]] to test the integrity of the [[mitochondrial outer membrane]] ([[cytochrome c control efficiency]]). +
The '''cytochrome ''c'' control efficiency''' expresses the control of respiration by externally added [[cytochrome c | cytochrome ''c'']], c, as a fractional change of flux from substrate state CHNO to CHNOc. These fluxes are corrected for ''Rox'' and may be measured in the OXPHOS state or ET state, but not in the LEAK state. In this [[flux control efficiency]], CHNOc is the [[reference state]] with stimulated flux; CHNO is the [[background state]] with CHNO substrates, upon which c is added:
''j''<sub>cyt ''c''</sub> = (''J''<sub>CHNOc</sub>-''J''<sub>CHNO</sub>)/''J''<sub>CHNOc</sub>. +
D
'''D number''' is the unique code given for each [[SUIT]] protocol. In the same [[MitoPedia: SUIT |SUIT protocol]] family (SUIT-###), there might be different protocols, specifically designed for different [[sample]] type (''e.g.'', different [[mitochondrial preparations]]) or for different applications (''e.g.'', O2, [[AmR]], [[Mitochondrial membrane potential|Fluo]], [[MgG]]). Since the use of different kinds of sample or application may result in slightly different steps, each protocol receives a different D-number. +
[[File:Dorabadge5.png|150px|right]] The Declaration on Research Assessment '''DORA''' recognizes the need to improve the ways in which researchers and the outputs of scholarly research are evaluated. +
'''DTPA''' (Diethylenetriamine-N,N,N',N,N-pentaacetic acid, pentetic acid,(Carboxymethyl)imino]bis(ethylenenitrilo)-tetra-acetic acid) is a polyaminopolycarboxylic acid (like EDTA) chelator of metal cations. DTPA wraps around a metal ion by forming up to eight bounds, because each COO- group and and N-center serves a center for chelation. With transition metals the number of bounds is less than eight. The compound is not cell membrane permeable. In general, it chelates multivalent ions stronger than EDTA. +
[[Image:Logo OROBOROS-DatLab.jpg|200px|right|DatLab]]'''DatLab''' is the O2k-Software for Data Acquisition & Analysis, specifically developed for [[high-resolution respirometry]] with the O2k.
The newest DatLab version is '''DatLab 8''', included in the O2k-Packages. NextGen-O2k and O2k-Series J* and higher come with DatLab 8 installed on the integrated PC (Linux). DatLab 8 is required for the NextGen-O2k. DatLab 8.1 is compatible with O2k-Series (E and higher).
The DatLab software is designed for 64-bit versions of Windows operating systems and does not run on MAC devices.
The minimum computer requirements are Intel-Core-2 or equivalent CPU, 2GB RAM and Windows XP (64-bit version). However, we recommend Intel i5 or equivalent CPU, 4GB RAM, Windows 10 (64-bit version) and SSD. For the proper display of DatLab on your computer, please make sure the “Language settings” are set to English.
*Optionally available without integrated PC. +
'''[[DatLab]] 2''' (DL2) is a MS-DOS programe. DL2 is still used for analysis of [[oxygen kinetics]], after exporting files recorded in recent DatLab versions. A user-friendly O2-kinetics module is in preparation (DL8). +
This is a brief summary of steps to be taken for performing a high-resolution respirometry experiment with '''[[SUIT protocols]]''' using the OROBOROS [[Oroboros O2k]] and '''[[DatLab]]''' software. (1) Search for a specific [[SUIT protocol name]] (go to [[MitoPedia:_SUIT#SUIT_protocols |MitoPedia: SUIT]]). The list of MitoPedia SUIT protocols can be sorted by [[categories of SUIT protocols]] (sorting by SUIT protocol name), which is listed as the 'abbreviation' of the SUIT protocol name. (2) Copy the template for [[Mark names]] into your DatLab subdirectory: DatLab\APPDATA\MTEMPLAT. (3) Copy the [[DatLab-Analysis templates |DatLab-Analysis template]] for this SUIT protocol. (4) Follow the link to the corresponding publication or MiPNet communication, where the pdf file describing the SUIT protocol is available. (5) A DatLab demo file may be available providing an experimental example. After each sequential titration, a mark is set on the plot for flux or flow. After having set all marks, pull down the 'Mark names' menu, select the corresponding SUIT protocol for mark names, and rename all marks. The Mark names template also provides standard values of the titration volume preceding each mark. (6) Go to 'Mark statistics' [F2], copy to clipboard, and paste into the sample tab in the DatLab-Analysis template.
: Example:
:* SUIT protocol name: [[SUIT-011]]
:* Mark names in DatLab: 1GM;2D;2c;3S;4U;5Rot-
:* DatLab-Analysis template: SUIT_NS(GM)01.xlsx
:* MiPNet communciation: [[MiPNet12.23 FibreRespiration]]
:* DatLab demo file: MiPNet12.23 FibreRespiration.DLD +
DatLab 8: The file type generated is *.dld8.
DatLab 7: The file type generated is *.DLD. +
Common '''DatLab error messages''' and according actions and solutions are listed here. +
[[Image:Logo OROBOROS-DatLab.jpg|left|60px||link=http://wiki.oroboros.at/index.php/DatLab |DatLab]]
We recommend a 'clean install' for '''DatLab installation''': rename your previous DatLab programme subdirectory (''e.g.'' C:\DatLab_OLD). The standard '''Instrumental and SUIT DL-Protocols''' package is automatically implemented with the simple DatLab programme installation. +
The quality of the results are strongly affected by the performance and data analysis. Therefore, we provide guidelines for performing and evaluating respirometric assays. +
'''DatLab templates''' can be imported for O2k-setups, graph layouts, mark names, TIP2k setups and marks statistics configurations.
:::: See also » [[Manage setups and templates - DatLab|Manage setups and templates]] +
Go in DatLab to [[Mark statistics - DatLab|Mark statistics]] (F2), select which type of marks you want to export ("All marks in plot" or "DL-Protocol marks", with 3 possibilities each), then click on [Copy to clipboard] to copy selected values and paste them to a '''DatLab-Analysis template''' for numerical and graphical data analysis. +
DatLab-Upgrading to DatLab 6: including free follow-up updates for DatLab 6 for the next two years +
'''DatLab-Upgrading\4.1-5.2''': Upgrading DatLab 4.x to 5.2, incl. O2k-Manual, with free follow-up updates of DatLab 5.2. '''Discontinued''': see higher [[DatLab]] version. +
The '''data recording interval''' is the time interval at which data is sampled with an instrument. In [[DatLab]] the data recording interval is set in the [[O2k control]] window. The system default value is 2 s. A lower data recording interval is selected for kinetic experiments, and when the volume-specific oxygen flux is high (>300 pmol O<sub>2</sub>·s<sup>-1</sup>·ml<sup>-1</sup>).<br/>Technically, the O2k instrument (hardware) measures the sensor signal every 10ms (which is NOT the „data recording interval“). By the given data recording interval from DatLab (software) a discrete number of sensor signal points are taken to calculate an average value in the O2k (e.g. a data recording interval of 2 s can take 200 sensor signal points; a data recording interval of 0.5 s can take 50 sensor signal points). This average value is sent to DatLab and is recorded as a raw data point. However, there is a defined threshold: the O2k does not apply more than 200 sensor signal points to calculate the average for the raw data point. For example a data recording interval of 3 s could take 300 sensor signal points but only the 200 most recent sensor signal points are used for averaging. +
'''DataCite''' is a global community of organizations and researchers identifying and citing research outputs and resources. We provide services to create persistent records of research, enable discovery and reuse, and support workflows throughout the research lifecycle. +
A '''dataset''' is a collection of data. In the context of databases a dataset represents the collection of entries in a database-table. In this table columns represent [[Attribute|attributes]] and rows display the according values of the entries. +
'''Dead cells''' dce are characterized by the loss of plasma membrane barrier function. The total cell count (''N''<sub>ce</sub>) is the sum of viable cells (''N''<sub>vce</sub>) and dead cells (''N''<sub>dce</sub>). +
A '''decimal marker''' is used to separate the integral part of numbers from the decimal part. The SI recommends: "the symbol for the decimal marker shall be either the point on the line or the comma on the line". In English language versions, the dot (point on the line) should be used uniquely as the decimal marker. To avoid ambiguities, BEC follows the SI recommendation that “Numbers may be divided in groups of three in order to facilitate reading; neither dots nor commas are ever inserted in the spaces between groups” (pages 183-184). +
The '''Default label''' is the system default value for the axis label. These labels are changed automatically, according to the selected channel and unit. To change this label enter a [[Custom label]]. +
Select '''Delete points''' in the [[Marks - DatLab |Mark information]] window to remove all data points in the marked section of the active plot. See also [[Interpolate points]] and [[Restore points]] or [[Recalculate slope]]. +
'''Density''', mass density ''ρ'' = ''m''·''V''<sup>-1</sup> [kg·m<sup>-3</sup>], is mass ''m'' divided by volume ''V''. Surface density ''ρ''<sub>A</sub> = ''m''·''A''<sup>-1</sup> [kg·m<sup>-2</sup>] ([[Bureau International des Poids et Mesures 2019 The International System of Units (SI) |SI]]). For a pure [[sample]] S, the mass density ''ρ''<sub>S</sub> = ''m''<sub>S</sub>·''V''<sub>S</sub><sup>-1</sup> [kg·m<sup>-3</sup>] is the [[mass]] ''m'' of pure sample S per [[volume]] ''V''<sub>S</sub> of the pure sample. With density ''ρ'' thus defined, the 'amount density' of substance B is ''ρ''<sub>B</sub> = ''n''<sub>B</sub>·''V''<sub>B</sub><sup>-1</sup> [mol·m<sup>-3</sup>]. This is not a commonly used expression, but the inverse is defined as the [[molar volume]] of a pure substance ([[Cohen 2008 IUPAC Green Book |IUPAC]]), ''V''<sub>m,B</sub> = ''V''<sub>B</sub>·''n''<sub>B</sub><sup>-1</sup> [m<sup>3</sup>·mol<sup>-1</sup>]. The pure sample is a pure gas, pure liquid or pure solid of a defined elementary entity. The amount [[concentration]], ''c''<sub>B</sub> = ''n''<sub>B</sub>·''V''<sup>-1</sup> [mol·m<sup>-3</sup>] is the amount ''n''<sub>B</sub> of substance B divided by the volume ''V'' of the mixture ([[Cohen 2008 IUPAC Green Book |IUPAC]]), and this is not called an 'amount density'. The term 'amount density' is reserved for an amount of substance per volume ''V''<sub>S</sub> of the pure substance. This explicit distinction between 'density' related to the volume of the ''sample'' and 'concentration' related to the total volume of the ''mixture'' is very helpful to avoid confusion. Further clarification is required in cases, when the mass density ''ρ''<sub>s</sub> of the sample in the mixture differs from the mass density ''ρ''<sub>S</sub> of the pure sample before mixing. Think of a sample S of pure ethanol with a volume of 1 L at 25 °C, which is mixed with a volume of 1 L of pure water at 25 °C: after the temperature of the mixture has equilibrated to 25 °C, the total volume of the mixture is less than 2 L, such that the volume ''V''<sub>S</sub> of 1 L pure ethanol has diminished to a smaller volume ''V''<sub>s</sub> of ethanol in the mixture; the density of ethanol in the mixture is higher than the density of pure ethanol (this is incomplete [[additivity]]). The volume ''V''<sub>s</sub> of sample s in a mixture is by definition smaller than the total volume ''V'' of the mixture. Sample volume ''V''<sub>S</sub> and system volume ''V'' are identical, but this applies only to the case of a ''pure'' sample. ''Concentration'' is related to samples s per total volume ''V'' of the mixture, whereas ''density'' is related to samples S or s per volume ''V''<sub>S</sub> = ''V'' or ''V''<sub>s</sub> < ''V'', respectively ([[BEC 2020.1]]).
'''Derivative spectroscopy''' can be used to eliminate interfering artefacts or species. A first order derivative will remove a constant background [[absorbance]] across the spectral range. A second order derivative spectrum will remove a species whose absorbance is linearly dependent upon the wavelength, etc.. +
[[O2k signals and output|Channels]] can be selected/deselected in [[DatLab]] in the [[O2k configuration]]. Deselect all O2k-MultiSensor channels in O2k-Core applications. Select only the specifically used channels in O2k-MultiSensor applications. +
A '''detector''' is a device that converts the light falling upon it into a current or voltage that is proportional to the light intensity. The most common devices in current use for [[fluorometry]] and [[spectrophotometry]] are [[photodiodes]] and [[photodiode arrays]]. +
'''Diapause''' is a preprogrammed form of developmental arrest that allows animals to survive harsh environmental conditions and may also allow populations to synchronize periods of growth and reproduction with periods of optimal temperatures and adequate water and food. Diapause is ''endogenously'' controlled, and this dormancy typically begins well before conditions become too harsh to support normal growth and development [1,2]. » [[Diapause#Diapause versus quiescence| '''MiPNet article''']] +
The '''dicarboxylate carrier''' is a transporter which catalyses the electroneutral exchange of [[malate]]<sup>2-</sup> (or [[succinate]]<sup>2-</sup>) for inorganic [[phosphate]], HPO<sub>4</sub><sup>2-</sup>. +
A '''difference spectrum''' is an [[absorbance spectrum]] obtained by subtracting that of one substance from that of another. For example, a '''difference spectrum''' may be plotted of the [[absorbance spectrum]] obtain ed from reduced [[cytochrome c]] and subtracting the [[absorbance spectrum]] from the same concentration of [[cytochrome c]] in its oxidised state. The [[difference spectrum]] may be used to quantify the amount to which the [[cytochrome c]] is reduced. This can be achieved with the aid of a [[reference spectrum]] (or spectra) and the [[least squares method]]. +
What are potential causes for '''different O<sub>2</sub> fluxes in the left and right chamber'''? +
'''Diffraction gratings''' are [[dispersion devices]] that are made from glass etched with fine grooves, spaced at the same order of magnitude as the wavelength of the light to be dispersed, and then coated with aluminium to reflect the light to the photodiode array. '''Diffraction gratings''' reflect the light in different orders and [[filters]] need to be incorporated to prevent overlapping. +
A '''Digital Object Identifier''', DOI, is a persistent identifier used to uniquely identify online publications in order to ensure they remain traceable, searchable and citable over the long term. Compared to other types of persistent identifiers, the DOI system is widespread and well established in the life sciences arena, and it provides widely accepted visible proof that a publication is citable. +
'''Digitonin''' is a mild detergent that permeabilizes plasma membranes selectively due to their high cholesterol content, whereas mt-membranes with lower cholesterol content are affected only at higher concentrations. Digitonin is a natural product and thus the effective concentration has to be determined by titrations for every batch. The optimum effective digitonin concentrations for complete plasma membrane permeabilization of cultured cells can be determined directly in a respirometric protocol (see: [[SUIT-010 O2 ce-pce D008]]). +
'''Dihydro-orotate dehydrogenase''' is an electron transfer complex of the inner mitochondrial membrane, converting dihydro-orotate (Dho) into orotate, and linking electron transfer through the [[Q-junction]] to pyrimidine synthesis and thus to the control of biogenesis. +
'''Dihydroethidium''' (also called hydroethidine) is a cell permeant fluorescent probe used to analyse superoxide presence. It is a reduced form of ethidium that presents blue fluorescence, and after oxidation by superoxide becomes able to intercalate DNA and emits red fluorescence (excitation wavelength ~518–535 nm, emission ~605–610 nm). It has been used to detect superoxide by HPLC and by fluorescence microscopy. +
Dilution of the concentration of a compound or sample in the experimental chamber by a titration of another solution into the chamber. +
'''Dimensions''' are defined in the SI {''Quote''}: Physical quantities can be organized in a system of dimensions, where the system used is decided by convention. Each of the seven base quantities used in the SI is regarded as having its own dimension. .. All other quantities, with the exception of [[count]]s, are derived quantities, which may be written in terms of base quantities according to the equations of physics. The dimensions of the derived quantities are written as products of powers of the dimensions of the base quantities using the equations that relate the derived quantities to the base quantities.
There are quantities ''Q'' for which the defining equation is such that all of the dimensional exponents in the equation for the dimension of ''Q'' are zero. This is true in particular for any quantity that is defined as the ratio of two quantities of the same kind. .. There are also some quantities that cannot be described in terms of the seven base quantities of the SI, but have the nature of a [[count]]. Examples are a number of molecules, a number of cellular or biomolecular entities (for example copies of a particular nucleic acid sequence), or degeneracy in quantum mechanics. Counting quantities are also quantities with the associated unit one. {''end of Quote'': p 136, [[Bureau International des Poids et Mesures 2019 The International System of Units (SI)]]} +
'''Dimethyl sulfoxide''' is a polar aprotic solvent that dissolves both polar and nonpolar compounds and is miscible in a wide range of organic solvents as well as water. DMSO may also be used as a cryoprotectant, added to cell media to reduce ice formation and thereby prevent cell death during the freezing process. +
'''Dinitrochlorobenzene (1-chloro-2,4-dinitrobenzene)''' (DNCB) is a glutathione (GSH) inhibitor. +
'''2,4-dinitrophenole''' (C<sub>6</sub>H<sub>4</sub>N<sub>2</sub>O<sub>5</sub>; M = 184.11 g·mol<sup>-1</sup>) is a protonophore acting as an [[uncoupler]] of [[oxidative phosphorylation]]. +
A '''directive''' is a legal act of the European Union, which requires member states to achieve a particular result without dictating the means of achieving that result. +
The '''Directory of Open Access Journals''' is a free online directory that indexes and provides access to open access peer-reviewed journals. +
In a '''discontinuous system''', gradients in [[continuous system]]s across the length, ''l'', of the diffusion path [m], are replaced by differences across compartmental boundaries of zero thickness, and the local concentration is replaced by the free activity, ''α'' [mol·dm<sup>-3</sup>]. The length of the diffusion path may not be constant along all diffusion pathways, spacial direction varies (''e.g.'', in a spherical particle surrounded by a semipermeable membrane), and information on the diffusion paths may even be not known in a discontinuous system. In this case (''e.g.'', in most treatments of the [[protonmotive force]]) the diffusion path is moved from the (ergodynamic) isomorphic [[force]] term to the (kinetic) [[mobility]] term. The synonym of a discontinuous system is '''compartmental''' or discretized system. In the first part of the definition of discontinuous systems, three compartments are considered: (1) the source compartment A, (2) the sink compartment B, and (3) the internal barrier compartment with thickness ''l''. In a two-compartmental description, a system boundary is defined of zero thickness, such that the barrier comparment (''e.g.'', a semipermeable membrane) is either part of the system (internal) or part of the environment (external). Similarly, the intermediary steps in a chemical reaction may be explicitely considered in an ergodnamic multi-comparment system; alternatively, the kinetic analysis of all intermediary steps may be collectively considered in the catalytic reaction ''mobility'', reducing the measurement to a two-compartmental analysis of the substrate and product compartments. +
A '''dispersion device''' diffracts light at different angles according to its wavelength. Traditionally, prisms and [[diffraction gratings]] are used, the latter now being the most commonly used device in a [[spectrofluorometer]] or [[spectrophotometer]]. +
'''Display DatLab help'''
In this section, we present some issues that could happen during your data analysis related to the graphs display and how to fix them quickly.
Case in which an issue might occur:
::* While analysing your data, trying to close the program while the graph is still being loaded. If you cancel the closing window, the graph will not be loaded at the screen.
In the event of a frozen display of the graphs, try the alternatives below:
::* Click on: Graph > Autoscale time axis
::* Click on: Graph > Scaling (F6); then press OK to confirm the scaling and induce the program to reload the graphs (no changes in the graphs are required). +
The Power-O2k number, which is set in the pull-down menu Oroboros O2k \ [[O2k configuration]], is shown in the active graph. To show it in graphs copied to clipboard, the option "Show Oroboros icon in clipboard files" must be enabled in the Graph-menu [[Graph options - DatLab]]. +
If '''Display numerical value''' the current numerical values are displayed in the graph for the active plots on the Y1 axis and Y2 axis (during data acquisition only). +
The sodium salt of '''Dithionite''' Na<sub>2</sub>S<sub>2</sub>O<sub>4</sub> (Dit) is the 'zero oxygen solution powder' used for [[Oxygen calibration - DatLab |calibration of oxygen sensors]] at [[Zero calibration | zero oxygen concentration]], or for stepwise reduction of oxygen [[concentration]]s in [[MiPNet14.06 Instrumental O2 background |instrumental O<sub>2</sub> background tests]]. It is not recommended to use dithionite in experiments with biological samples or several multisensor approaches, for these see [[Setting the oxygen concentration]]. +
The most common cause of '''drift''' is variation in the intensity of the [[light source]]. The effect of this can be minimised by carrying out a [[balance]] at frequent intervals. +
If a sample contains a number of absorbing substances, it is sometimes possible to select discrete pairs of wavelengths at which the change in [[absorbance]] of a particular substance (due to oxidation or reduction, for example) is largely independent of changes in the [[absorbance]] of other substances present. '''Dual wavelength analysis''' can be carried out for [[cytochrome c]] by subtracting the [[absorbance]] at 540 nm from that at 550nm in order to give a measure of the degree of reduction. Similarly, by subtracting the [[absorbance]] at 465 nm from that at 444 nm, an indicator of the [[redox state]] of [[Complex IV | cytochrome ''aa''<sub>3</sub>]] can be obtained. +
[[Electron-transfer-pathway state |ET-pathway level 2]] is supported by '''duroquinol''' DQ feeding electrons into Complex III (CIII) with further electron transfer to CIV and oxygen. Upstream pathways are inhibited by rotenone and malonic acid in the absence of other substrates linked to ET-pathways with entry into the Q-junction. +
'''Dyscoupled respiration''' is [[LEAK respiration]] distinguished from intrinsically (physiologically) uncoupled and from extrinsic experimentally [[Uncoupler|uncoupled]] respiration as an indication of extrinsic uncoupling (pathological, toxicological, pharmacological by agents that are not specifically applied to induce uncoupling, but are tested for their potential dyscoupling effect). Dyscoupling indicates a mitochondrial dysfunction.
In addition to intrinsic uncoupling, dyscoupling occurs under pathological and toxicological conditions. Thus a distinction is made between physiological uncoupling and pathologically defective dyscoupling in mitochondrial respiration. +
E
» [[Energy]], [[Exergy]] ''E''
» [[elementary charge]] ''e'' = 1.602 176 634∙10<sup>-19</sup> C∙x<sup>-1</sup>
» [[Euler's number]] ''e'' ~ 2.718 281 828 459
» [[ET capacity]] ''E'' +
[[File:J(E-L).jpg|50 px|E-L coupling efficiency]] The '''''E-L'' coupling efficiency''', ''j<sub>E-L</sub>'' = (''E-L'')/''E'' = 1-''L/E'', is 0.0 at zero coupling (''L''=''E'') and 1.0 at the limit of a fully coupled system (''L''=0). The background state is the [[LEAK respiration|LEAK]] state which is stimulated to flux in the [[electron transfer pathway]] reference state by [[uncoupler]] titration. LEAK states ''L''<sub>N</sub> or ''L''<sub>T</sub> may be stimulated first by saturating ADP (rate ''P'' in the OXPHOS state) with subsequent uncoupler titration to the ET state with maximum rate ''E''. The ''E-L'' coupling efficiency is based on measurement of a [[coupling-control ratio]] ([[LEAK-control ratio]], ''L/E''), whereas the thermodynamic or [[ergodynamic efficiency]] of coupling between ATP production (phosphorylation of ADP to ATP) and oxygen consumption is based on measurement of the output/input flux ratio (P»/O<sub>2</sub> ratio) and output/input force ratio (Gibbs force of phosphorylation/Gibbs force of oxidation). The [[biochemical coupling efficiency]] expressed as the ''E-L'' coupling efficiency is independent of kinetic control by the ''E-P'' control efficiency, and is equal to the [[P-L control efficiency |''P-L'' control efficiency]] if ''P=E'' as evaluated in a [[coupling-control protocol]].
» [[#Biochemical_coupling_efficiency:_from_0_to_.3C1 | '''MiPNet article''']] +
[[Image:E-L.jpg|50 px|E-L net ET capacity]] The '''''E-L'' net ET capacity''' is the [[ET capacity]] corrected for [[LEAK respiration]]. ''E-L'' is the respiratory capacity potentially available for ion transport and phosphorylation of ADP to ATP. Oxygen consumption in the ET-pathway state, therefore, is partitioned into the ''E-L'' net ET capacity and LEAK respiration ''L<sub>P</sub>'', compensating for proton leaks, slip and cation cycling: ''E'' = ''E-L''+''L<sub>P</sub>'' (see [[P-L net OXPHOS capacity]]). +
[[File:J(E-P).jpg|50 px|E-P control efficiency]] The '''''E-P'' control efficiency''', ''j<sub>E-P</sub>'' = (''E-P'')/''E'' = 1-''P/E'', is an expression of the relative limitation of [[OXPHOS capacity]] by the capacity of the [[phosphorylation system]]. It is the normalized ''E-P'' excess capacity. ''j<sub>E-P</sub>'' = 0.0 when OXPHOS capacity is not limited by the phosphorylation system at zero ''E-P'' excess capacity, ''P''=''E'', when the phosphorylation system does not exert any control over OXPHOS capacity. ''j<sub>E-P</sub>'' increases with increasing control of the phosphorylation system over OXPHOS capacity. ''j<sub>E-P</sub>'' = 1 at the limit of zero phosphorylation capacity. The [[OXPHOS]] state of mt-preparations is stimulated to [[electron transfer pathway]] capacity ''E'' by [[uncoupler]] titration, which yields the [[E-P excess capacity |''E-P'' excess capacity]]. +
[[Image:ExP.jpg|60 px|link=E-P excess capacity|''E-P'' excess capacity]] The '''''E-P'' excess capacity''' is the difference of the [[ET capacity]] and [[OXPHOS capacity]]. At ''E-P'' > 0, the capacity of the [[phosphorylation system]] exerts a limiting effect on OXPHOS capacity. In addition, ''E-P'' depends on coupling efficiency, since ''P'' aproaches ''E'' at increasing uncoupling. +
[[Image:j(E-R).jpg|50 px|E-R control efficiency]] The '''''E-R'' control efficiency''', ''j<sub>E-R</sub>'' = (''E-R'')/''E'' = 1-''R/E'', is an expression of the relative scope of increasing [[ROUTINE respiration]] in living cells by uncoupler titrations to obtain [[ET capacity]]. ''j<sub>E-R</sub>'' = 0.0 for zero ''E-R'' reserve capacity when ''R''=''E''; ''j<sub>E-R</sub>'' = 1.0 for the maximum limit when ''R''=0. The [[ROUTINE]] state of living cells is stimulated to [[electron transfer pathway]] capacity by [[uncoupler]] titration, which yields the [[E-R reserve capacity |''E-R'' reserve capacity]]. Since ET capacity is significantly higher than [[OXPHOS capacity]] in various cell types (as shown by '''[[cell ergometry]]'''), ''j<sub>E-R</sub>'' is not a reserve capacity available for the cell to increase oxidative phosphorylation, but strictly a scope (reserve) for uncoupling respiration. Similarly, the apparent [[E-P excess ET capacity |''E-P'' excess ET capacity]] is not a respiratory reserve in the sense of oxidative phosphorylation. +
[[Image:ExR.jpg|60 px|E-R reserve capacity]] The '''''E-R'' reserve capacity''' is the difference of [[ET capacity]] and [[ROUTINE respiration]]. For further information, see [[Cell ergometry]]. +
[[File:E.jpg]] '''T capacity''' is the respiratory electron-transfer-pathway capacity ''E'' of mitochondria measured as oxygen consumption in the noncoupled state at optimum [[uncoupler]] concentration. This optimum concentration is obtained by stepwise titration of an established protonophore to induce maximum oxygen flux as the determinant of ET capacity. The experimentally induced noncoupled state at optimum uncoupler concentration is thus distinguished from (''1'') a wide range of uncoupled states at any experimental uncoupler concentration, (''2'') physiological uncoupled states controlled by intrinsic uncoupling (e.g. UCP1 in brown fat), and (''3'') pathological dyscoupled states indicative of mitochondrial injuries or toxic effects of pharmacological or environmental substances. ET capacity in mitochondrial preparations requires the addition of defined fuel substrates to establish an ET-pathway competent state.
» [[#Why ET capacity, why not State 3u.3F | '''MiPNet article''']] +
[[Electron transfer pathway]] competent state, ''see'' '''[[Electron-transfer-pathway state]]'''. +
See '''[[Electron-transfer-pathway state]]''' +
[[File:EUROMIT.jpg|left|250px]]
'''EUROMIT''' is a group based in Europe for organizing '''International Meetings on Mitochondrial Pathology'''. +
'''Ectotherms''' are organisms whose body temperatures conform to the thermal environment. In many cases, therefore, ectotherms are [[poicilotherms | poicilothermic]]. +
'''Editorial board participation''' is a topic addressed in [[COPE core practices for research]]. +
'''Bendavia''' ('''Elamipretide''') was developed as a mitochondria-targeted drug against degenerative diseases, including cardiac ischemia-reperfusion injury. Clinical trials showed variable results. It is a cationic tetrapeptide which readily passes cell membranes, associates with [[cardiolipin]] in the mitochondrial inner membrane. Supercomplex-associated CIV activity significantly improved in response to elamipretide treatment in the failing human heart. +
According to David Fell, "Elasticities are properties of individual enzymes and not the metabolic system. The elasticity of an enzyme to a metabolite is related to the slope of the curve of the enzyme's rate plotted against metabolite concentration, taken at the metabolite concentrations found in the pathway in the metabolic state of interest. It can be obtained directly as the slope of the logarithm of the rate plotted against the logarithm of the metabolic concentration. The elasticity will change at each point of the curve (s,v) and must be calculated for the specific concentration of the metabolite (s) that will give a specific rate (r) of the enzyme activity" (See Figure).
[[File:Elasticity_Measurement.jpg]] +
'''Current''' or electric [[flow]] ''I''<sub>el</sub> is the [[advancement]] of [[charge]] per unit of time, expressed in the SI base unit [[ampere]] [C·s<sup>-1</sup> = A]. Electrons or ions are the current-carrying [[motive entity |motive entities]] of electric flow. Electrons e<sup>-</sup> are negatively charged subatomic particles carrying 'negative electricity' with a mass that is about 1/1700 of the smallest particle — the proton — carrying 'positive electricity' (Thompson 1906). Correspondingly the [[velocity]] of electrons is much higher than that of protons or any other (larger) ion. Current is the velocity ''v'' of paticles times the number of motive charges. Therefore, electron current ''I''<sub>e<sup>-</sup></sub> is of a different nature from electric current ''I''<sub>el''χ''</sub> carried by all species ''i'' of ions ''X<sub>i</sub>'' (cations and anions) summarized as ''χ'' = Σ(''z<sub>i</sub>''·''X<sub>i</sub>''). Whereas ''I''<sub>e<sup>-</sup></sub> is the net translocation of electrons moving forwards and backwards, ''I''<sub>el''χ''</sub> is the net translocation of charges carried by different cations and anions. In contrast, ion current ''I''<sub>elX</sub> of a specific ion X is the partial translocation of charges carried by net translocation of ion X only. If cation current ''I''<sub>elX<sup>+</sup></sub> is antagonized entirely by counterion current ''I''<sub>elY<sup>-</sup></sub> as the process of antiport, then the electric current ''I''<sub>el''χ''</sub> is zero. The (net) electric current in a compartmental system is driven by the electric force Δ<sub>el</sub>''F''<sub>p<sup>+</sup></sub> or electric potential difference Δ''Ψ''<sub>p<sup>+</sup></sub>, whereas a compensated ion/counterion antiport current is insensitive to the electric potential difference. +
'''Electric current density''' is [[current]] divided by area, ''j''=''I''·''A''<sup>-1</sup> [C·m<sup>-2</sup>]. Compare: [[density]]. +
[[File:Table Physical constants.png|right|400px|thumb|]] The '''electrochemical constant''' ''f'' has the SI unit for energy per charge per temperature [J·C<sup>-1</sup>·K<sup>-1</sup>].
''f'' = ''k''·''e''<sup>-1</sup>, the [[Boltzmann constant]] ''k'' divided by the [[elementary charge]] ''e''.
''f'' = ''R''·''F''<sup>-1</sup>, the [[gas constant]] ''R'' divided by the [[Faraday constant]] ''F''. +
[[Image:Electrolyte Reference-Electrode.jpg|right|180px|link=http://www.bioblast.at/index.php/Electrolyte%5CReference-Electrode]]'''Electrolyte\Reference-Electrode''' for [[Reference-Electrode\2.4 mm]] +
'''Electron flow''' through the mitochondrial [[Electron transfer pathway]] (ET-pahway) is the scalar component of chemical reactions in oxidative phosphorylation ([[OXPHOS]]). Electron flow is most conveniently measured as oxygen consumption (oxygraphic measurement of [[oxygen flow]]), with four electrons being taken up when oxygen (O<sub>2</sub>) is reduced to water. +
Electrons that escape the [[electron transfer pathway]] without completing the reduction of oxygen to water at cytochrome ''c'' oxidase, causing the production of [[Reactive_oxygen_species |ROS]]. The rate of electron leak depends on the topology of the complex, the redox state of the moiety responsible of electron leakiness and usually on the protonmotive force ([[Protonmotive force|Δ''p'']]). In some cases, the [[Protonmotive force|Δ''p'']] dependance relies more on the ∆pH component than in the ∆''Ψ''. +
In the mitochondrial '''electron transfer pathway''' (ET pathway) electrons are transferred from externally supplied reduced fuel substrates to oxygen. Based on this experimentally oriented definition (see [[ET capacity]]), the ET pathway consists of (1) the [[membrane-bound ET pathway]] with respiratory complexes located in the inner mt-membrane, (2) [[TCA cycle]] and other mt-matrix dehydrogenases generating NADH and succinate, and (3) the carriers involved in metabolite transport across the mt-membranes.
» [[#Electron transfer pathway versus electron transport chain |'''MiPNet article''']] +
[[File:SUIT-catg FNSGpCIV.jpg|right|400px]]
'''Electron-transfer-pathway states''' are obtained in [[mitochondrial preparations]] (isolated mitochondria, permeabilized cells, permeabilized tissues, tissue homogenate) by depletion of endogenous substrates and addition to the mitochondrial respiration medium of fuel substrates (CHNO) activating specific mitochondrial pathways, and possibly inhibitors of specific pathways. Mitochondrial electron-transfer-pathway states have to be defined complementary to mitochondrial [[coupling-control state]]s. [[Coupling-control state]]s require [[Electron-transfer-pathway state|ET-pathway competent states]], including oxygen supply. [[Categories of SUIT protocols]] are defined according to mitochondrial ET-pathway states.
» [[#ET_pathway_states |'''MiPNet article''']] +
'''Electron-transferring flavoprotein Complex''' (CETF) is a respiratory Complex localized at the matrix face of the inner mitochondrial membrane, supplies electrons to Q, and is thus an enzyme Complex of the mitochondrial [[Electron transfer pathway]] (ET-pathway). CETF links the ß-oxidation cycle with the membrane-bound electron transfer system in [[fatty acid oxidation]] (FAO). +
'''Electronic-TIP2k Upgrading\O2k-Main Unit Series A-D - Former Product ''': not required for [[O2k-Core]], the [[O2k-Main Unit]] has to be returned to the OROBOROS workshop. +
'''Electronic-TIP2k Upgrading\O2k-Main Unit Series E - Former Series ''': not required for [[O2k-Core]], free of charge for Series E in conjunction with the purchase of the [[TIP2k-Module]], the [[O2k-Main Unit]] has to be returned to the OROBOROS workshop. +
[[File:Table Physical constants.png|right|400px|thumb|]] The '''elementary charge''' or proton charge ''e'' has the SI unit coulomb [C], but more strictly coulomb per elementary unit [C·x<sup>-1</sup>]. -''e'' is the charge per electron. Elementary charge ''e'' is the charge per [[elementary entity]] H<sup>+</sup> with SI unit [C] but canonical SI unit [C·x<sup>-1</sup>]. When the charge ''Q''<sub>el</sub> [C] of a number ''N''<sub>e</sub> [x] of electrons e is divided by the count ''N''<sub>e</sub>, then the [[particle charge]] ''Q<sub>N<sub>X</sub></sub>'' (''Q<sub><u>N</u>X</sub>'') charge per elementary entity is obtained, -''e'' = ''Q''<sub>el</sub>/''N''<sub>e</sub> [C·x<sup>-1</sup>]. ''e'' is also used as an atomic unit. +
[[File:Count-vs-number.png|right|120px|link=Unit]]
An '''elementary entity''' is an [[entity]] of type ''X'', distinguished as a single ''[[unit]]'' of countable objects (''X'' = molecules, cells, organisms, particles, parties, items) or events (''X'' = beats, collisions, emissions, decays, celestial cycles, instances, occurrences, parties). "An elementary entity may be an atom, a molecule, an ion, an electron, any other particle or specified group of particles" ([[Bureau International des Poids et Mesures 2019 The International System of Units (SI) |Bureau International des Poids et Mesures 2019)]]. An elementary entity, therefore, needs to be distinguished from non-countable entities and the general class of entities ''X''. This distinction is emphasized by the term 'elementary' (synonymous with 'elementary entity') with symbol ''U''<sub>''X''</sub> and [[unit |elementary unit]] [x].
If an object is defined as an assembly of particles (a party of two, a molecule as the assembly of a stoichiometric number of atoms), then the elementary is the assembly but not the assembled particle. A number of defined elementaries ''U''<sub>''X''</sub> is a [[count]], ''N''<sub>''X''</sub> = ''N''·''U''<sub>''X''</sub> [x], where ''N'' is a number, and as such ''N'' is dimensionless, and ''N'' is a ''number'' (stop) and is not 'a number of ..'. Elementaries are added as items to a count. The elementary ''U''<sub>''X''</sub> has the [[dimension]] U of the [[count]] ''N''<sub>''X''</sub>. The elementary ''U''<sub>''X''</sub> has the same unit [x] as the count ''N''<sub>''X''</sub>, or more accurately it gives the count the defining 'counting-unit', which is the 'elementary unit' [x]. From the definition of count as the number (''N'') of elementaries (''U'') of entity type ''X'', it follows that count divided by elementary is a pure number, ''N'' = ''N''<sub>''X''</sub>·''U''<sub>''X''</sub><sup>-1</sup>. The unit x of a count can neither be the entity ''X'' nor a number. The elementary of type ''X'' defines the identity ''X'' of the elementary ''U''<sub>''X''</sub> with the unit 'elementary unit' with symbol [x]. Since a count ''N''<sub>''X''</sub> is the number of elementary entities, the elementary ''U''<sub>''X''</sub> is not a count (''U''<sub>''X''</sub> is not identical with ''N''·''U''<sub>''X''</sub>).
[[File:Count-vs-number.png|right|120px|link=Elementary entity]]The '''elementary unit''' [x] is the unit of a [[count]] ''N''<sub>''X''</sub> [x]. The [[International System of Units]] defines the unit of a count as 1. Then the '''N'''umber 1 is the '''U'''nit of the '''C'''ount of '''E'''ntities — NUCE. This causes a number of formal inconsistencies which are resolved by introducing the elementary unit [x] as the abstracted unit of Euclid’s unit, which is an [[elementary entity]] ''U''<sub>''X''</sub> [x], and as the unit of Euclid’s number, which is a count ''N''<sub>''X''</sub> [x]. +
'''Enable DL-Protocol editing''' is a novel function of DatLab 7.4 offering a new feature in DL-Protocols: flexibility. Fixed sequences of events and marks can be changed (Skip/Added) in a SUIT protocol by the user. Moreover, the text, instructions, concentrations and titration volumes of injections in a specific DL-Protocol can be edited and saved as [[Export_DL-Protocol_User_(*.DLPU)| user-specific DL-Protocol]] [File]\Export\DL-Protocol User (*.DLPU). To enable it, under the 'Protocols' tab in the menu, select the option 'Enable DL-Protocol editing', and then select the plot in which the marks will be set (''e.g.,'' O2 flux per V). Select the 'Overview' window, where you will be able to edit events and marks names, definition/state, final concentration and titration volumes, as well as select a mark as 'multi' for multiple titration steps, skip a mark, or add a new event or mark. After saving, [[Export_DL-Protocol_User_(*.DLPU)|export a DL-Protocol User (DLPU)]] and load it before running the next experiments. If users of DatLab versions older than DatLab 7.4 wish to alter the nature of the chemicals used or the sequence of injections, we ask them to [https://www.oroboros.at/index.php/o2k-technical-support/ contact the O2k-Technical Support].
For more information:
[[Image:PlayVideo.jpg|50px|link=https://www.youtube.com/watch?v=Vd66dHx-MyI]] [https://www.youtube.com/watch?v=Vd66dHx-MyI Export DL-Protocol User (*.DLPU)] +
'''Endergonic''' transformations or processes can proceed in the forward direction only by coupling to an [[exergonic]] process with a driving force more negative than the positive force of the endergonic process. The backward direction of an endergonic process is exergonic. The distinction between endergonic and [[endothermic]] processes is at the heart of [[ergodynamics]], emphasising the concept of [[exergy]] changes, linked to the performance of [[work]], in contrast to [[enthalpy]] changes, linked to [[heat]] or thermal processes, the latter expression being terminologically linked to ''thermodynamics''. +
