Category:NextGen-O2k

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NextGen-O2k


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Category:NextGen-O2k

The revolutionary all-in-one instrument to conquer mitochondrial disease

NextGen-O2k

MitoPedia: NextGen-O2k

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TermAbbreviationDescription
Hydrogen peroxideH2O2
Hydrogen peroxide
Hydrogen peroxide, H2O2 or dihydrogen dioxide, is one of several reactive oxygen intermediates generally referred to as reactive oxygen species (ROS). It is formed in various enzyme-catalyzed reactions (e.g., superoxide dismutase) with the potential to damage cellular molecules and structures. H2O2 is dismutated by catalase to water and oxygen. H2O2 is produced as a signaling molecule in aerobic metabolism and passes membranes more easily compared to other ROS.
Light-enhanced dark respirationLEDRLight-enhanced dark respiration LEDR is a sharp (negative) maximum of dark respiration in plants in response to illumination, measured immediately after switching off the light. LEDR is supported by respiratory substrates produced during photosynthesis and closely reflects light-enhanced photorespiration (Xue et al 1996). Based on this assumption, the the total photosynthetic oxygen flux TP is calculated as the sum of the measured net photosynthetic oxygen flux NP plus the absolute value of LEDR.
Mitochondrial membrane potentialmtMP, Δψ [V]The mitochondrial membrane potential, mtMP, is the electric part of the protonmotive force, ΔpH+.

Δψ = ΔpH+ - ΔµH+ / F

mtMP or Δψ is the potential difference across the inner mitochondrial (mt) membrane, expressed in the electric unit of volt [V]. Electric force of the mitochondrial membrane potential is the electric energy change per ‘motive’ electron or per electron moved across the transmembrane potential difference, with the number of ‘motive’ electrons expressed in the unit coulomb [C].
NADH-ModuleNADH-ModuleThe NADH-Module, is a component of the NextGen-O2k for simultaneous measurement of oxygen consumption and NAD(P)H autofluorescence. NAD(P)H autofluorescence is used to evaluate the redox state of the NAD(P)H-pool. The NADH-Module incorporates an UV light and NADH-Sensors which include a photodiode and specific filters.
NADH-SensorThe NADH-Sensor has been developed as a part of the NADH-Module for simultaneous monitoring of oxygen consumption and NADH redox state. The NADH-Sensor is composed of a photodiode and equipped with supergel R370 Italian blue filters (Rosco, US).
NextGen-O2k
NextGen-O2k InstrumentNextGen-O2k Instrument
NextGen-O2k Technical developments
Oxygen kineticsOxygen kinetics describes the dependence of respiration of isolated mitochondria or cells on oxygen partial pressure. Frequently, a strictly hyperbolic kinetics is observed, with two parameters, the oxygen pressure at half-maximum flux, p50, and maximum flux, Jmax. The p50 is in the range of 0.2 to 0.8 kPa for cytochrome c oxidase, isolated mitochondria and small cells, strongly dependent on Jmax and coupling state.
PB-ModulePB-ModuleThe PB-Module has been developed for conducting measurements of PhotoBiology, including photosynthesis. It consists of the PB-Sensor and electronic components which are an integral part of the NextGen-O2k. Measurements are recorded and evaluated with the DatLab 8 software.
PB-SensorThe PhotoBiology Light Source (PBLS) has been designed as a part of the PB-Module to provide with an external source of light. This enables experiments for evaluating the production of O2 in the presence of light. The PBLS consists of one LED and one photodiode mounted on the PBLS head protected by a PMR plastic cover. Three pairs of PBLS are provided with the PB-Module: white PBLS with an emitting wavelenght range of 425-750nm with two peaks at 440 and 630nm, blue PBLS with an emitting wavelength range of 439-457 nm and the peak at 451 nm, red PBLS with an emitting wavelength range of 620-632 nm and the peak at 634 nm. The light intensity can be regulated from 0 to 2750 µmol*s-1*m-2 (red PBLS), from 0 to 3000 µmol*s-1*m-2 (blue PBLS), from 0 to 3500 µmol*s-1*m-2 (white PBLS). The photodiode provides a real-time correction of the emitted light intensity to ensure a constant value throughout the experiment.
PhotoBiologyPBPhotoBiology is the science of the effect of light on biological processes. This includes photosynthesis, photochemistry, photophysics, photomorphogenesis, vision, bioluminescence, circadian rhythms and photodynamic therapy. Phototoxicity results from non-ionizing radiation (i.e. ultraviolet, visible and infrared radiation). Non-ionizing radiation is any type of electromagnetic radiation that does not carry enough energy per quantum (photon energy below 10 eV) to completely remove an electron from an atom or molecule. When photons interact with molecules, the molecules can absorb the photon energy and become excited, reacting with surrounding molecules and stimulating "photochemical" and "photophysical" changes. Respiration may be affected by light during photosynthesis or in dark respiration, with the transient response of light-enhanced dark respiration.
PhotosynthesisPSPhotosynthesis is the process that converts light energy into chemical energy which is subsequently transformed to the physiological energy demand. Photosynthesis has a light-dependent and light-independent (dark) phase. In plants, algae, and cynobacteria, light energy is absorbed during the light phase by the pigment chlorophyll and used to split water and generate adenosine triphosphate (ATP) and reducing power - nicotinamide adenine dinucleotide phosphate (NADPH), with the net production of O2 as a waste product. During the dark phase ATP and NADPH are used to synthesize carbohydrates from CO2 through the metabolic pathway called Calvin-Benson cycle. Oxygenic photosynthesis is responsible for producing and maintaining the oxygen concentration of the Earth’s atmosphere. In bacteria such as cyanobacteria, photosynthesis involves the plasma membrane and the cytoplasm. In eukaryotic cells (plants and algae), photosynthesis takes place in the chloroplasts.
Q-ModuleQ-ModuleThe Q-Module, developed both for measuring the Q redox state and for cyclic voltammetry measurements, is an integral part of the NextGen-O2k and consists of the Q-Sensor, integrated electronic components in the O2k, and the DatLab software.
Q-SensorThe Q-Sensor has been designed as a part of the Q-Module for measurements with cyclic voltammetry and voltammetry, allowing for analysis of the Q redox state. The Q-Stopper with the reference electrode is called Q-Sensor, which is plugged in the NextGen-O2k. A three-electrode system is used to detect the Q redox state. Two of the three electrodes (glassy carbon and platinum electrode) are built into the Q-Stopper, while the reference electrode is removable (Reference-Electrode\2.4 mm).
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Mitochondrial NADH and respiration

» Nicotinamide adenine dinucleotide
» NADH electron transfer-pathway state
» N-junction
» Complex I
» Rotenone
» Amytal
» Piericidin

Mitochondrial NADH production

» Tricarboxylic acid cycle
» Fatty acid oxidation

NextGen-O2k

» NextGen-O2k




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Phosphorylation pathway

» Phosphorylation pathway
» ATP synthase
» Adenine nucleotide translocase
» Phosphate carrier

Phosphorylation pathway substrates

» ADP
» ATP
» Inorganic phosphate

Phosphorylation pathway inhibitors

» Oligomycin
» Carboxyatractyloside
» Atractyloside
» Bongkrekik acid

Coupling control

» Oxidative phosphorylation
» OXPHOS capacity
» P-L control efficiency
» ROUTINE respiration
» R-L control efficiency

Respiratory complexes and coupling

» Complex I
» Complex III
» Complex IV
» Proton pump
» Electron-transfer-pathway state

ATP production measurement

» Magnesium Green



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PhotoBiology: photosynthesis

» PhotoBiology
» Photosynthesis

Plant physiology: respiration

» Photorespiration
» Chlororespiration
» Light-enhanced dark respiration

NextGen-O2k and PB-Module

» NextGen-O2k
» PB-Module
» PB-Sensor


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