Bioblast quiz

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Exemplary quiz

Note: Questions in this exemplary quiz were used from a set of questions prepared for the MiPschool Tromso-Bergen 2018: The protonmotive force and respiratory control. 1. Coupling of electron transfer reactions to vectorial translocation of protons. 2. From Einstein’s diffusion equation on gradients to Fick’s law on compartments. - Gnaiger 2018 MiPschool Tromso A2

Only one correct answer.

1 Convert the molar format of the Gibbs force of reaction, Δ_kF_nO2 [kJ/mol], into the electrical format, Δ_kF_eO2 [V]. Which physicochemical constant is required?

constant

2 What is the meaning of the symbol z_O2?

	Charge number of O2 = 4
	Alphabetical order of O2 isotope
	Atomic number of O2 = 8
	Elementary charge of O2 in [C]

3 How are the units of electric energy [J] and electric force [V] related?

	V = J/C
	V = J·C
	V = (J·F)/C
	V = J/(C·F)

4 Express -460 kJ/mol O₂ as electrical force in units of volt [V].

	1.2 V
	-1.2 V
	- 120 V
	- 1.2 kV

5 Why should we do that?

	To express both in identical motive units [MU]
	To compliment our brain mitochondria
	To feel insecure
	To get free drinks

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Blue Book Bioblast Quiz

Blue Book chapter 1: basic questions

1 The Oroboros-O2k is primarily designed for which type of research?

	Comprehensive mitochondrial function assessment, including oxygen consumption
	Glycolysis rate measurement
	Quantification of mitochondrial DNA
	Measurement of mitochondrial membrane potential only

2 Peter Mitchell's chemiosmotic coupling theory places fundamental importance on what concept for bioenergetics?

	The role of cytochromes
	Bioblasts as the systematic unit
	Mitochondrial DNA's function
	The operation of ATP synthase

3 Which is NOT a parameter measured by integrating fluorometry into high-resolution respirometry?

	H2O2 production
	Mitochondrial membrane potential changes
	Glucose uptake rates
	O2 consumption rates

4 The statement that mitochondrial fitness "solely depends on the genetic makeup of the individual" is:

	True, but only in the context of mitochondrial diseases.
	True, genetics are the only factor.
	Misleading, since mitochondrial fitness can be improved with supplements.
	Incorrect, as lifestyle and environmental factors also significantly influence mitochondrial fitness.

5 What does the term "bioblasts" refer to in the context of mitochondrial physiology?

	Elementary units or microorganisms acting wherever living forces are present, essentially mitochondria.
	The smallest units of DNA within mitochondria.
	A specific type of mitochondria found in muscle cells.
	Enzymes involved in the electron transport chain.

6 Which of the following is NOT a result of a measurement by the Oroboros-O2k?

	ATP production
	Calcium concentration
	H2O2 production
	Protein synthesis rates

7 What components constitute the protonmotive force (pmF) essential for ATP synthesis in mitochondria?

	ΔΨ and solute concentration
	Only ΔpH
	ΔΨ (mitochondrial membrane potential) and ΔpH
	Only ΔΨ

8 High-resolution respirometry (HRR) is primarily used for what purpose?

	Observing mitochondria physically
	Measuring cellular glucose concentration
	Quantitative analysis of mitochondrial respiration and function
	pH measurement of the mitochondrial matrix

9 Oxygen concentration impacts mitochondrial respiratory control by:

	Directly determining the rate of glycolysis
	Being inversely proportional to the rate of ATP synthesis
	Influencing exergonic and endergonic reactions in OXPHOS
	Having no significant impact on mitochondrial function

10 The "Q-junction" in mitochondrial respiratory control serves as:

	The location where glucose is converted into pyruvate
	A convergence point for multiple electron transport pathways
	The mitochondrial DNA replication site
	The site of ATP synthesis

11 SUIT protocols in mitochondrial research are designed to:

	Identify the best culture medium for mitochondrial growth
	Disrupt mitochondrial DNA and study its effects on respiration
	Analyze the effects of substrates, uncouplers, and inhibitors on respiratory control
	Measure the physical size of mitochondria under different conditions

12 NADH-linked substrates are used in physiological respiratory states to:

	Bypass the electron transport system
	Reflect the exclusive type of substrates used by mitochondria
	Demonstrate substrates irrelevant to mitochondrial physiology
	Represent substrates feeding electrons into the ETS, simulating physiological conditions

13 The primary purpose of integrating fluorometry with high-resolution respirometry is to:

	Increase the resolution of respirometry measurements alone
	Decrease the time required for each measurement
	Allow for the observation of mitochondrial shape and size
	Enable simultaneous measurement of oxygen consumption and other mitochondrial parameters

14 Which statement accurately describes the significance of LEAK respiration in the context of mitochondrial function?

	It denotes the respiration process exclusive to glycolytic cells.
	It represents the energy consumed to maintain ionic gradients in the absence of ATP synthesis.
	It is the maximum respiration rate achievable by mitochondria.
	It indicates the rate of oxygen consumption for ATP synthesis.

15 In mitochondrial research, the term "ET capacity" refers to:

	The capacity for energy transfer within the mitochondrion.
	The ability of the endoplasmic reticulum to transfer proteins to mitochondria.
	The maximum electron transport rate through the electron transport chain under optimal conditions.
	The enzyme titration capacity in metabolic pathways.

16 Which of the following is NOT a direct measurement capability of the Oroboros-O2k?

	Calcium ion concentration in the mitochondrial matrix
	Reactive oxygen species (ROS) production
	ATP production rates
	Mitochondrial DNA replication rates

17 The addition of fluorescent dyes in Oroboros-O2k and NextGen-O2k measurements allows for the assessment of:

	The rate of glycolysis in mitochondria
	Membrane fluidity and viscosity
	Nuclear DNA mutations
	Mitochondrial membrane potential changes

18 The primary purpose of substrate-uncoupler-inhibitor titration (SUIT) protocols in mitochondrial research is to:

	Identify the optimal conditions for ATP synthesis
	Measure the physical dimensions of mitochondria under various metabolic conditions
	Investigate the effects of different substrates, uncouplers, and inhibitors on mitochondrial respiratory control
	Determine the maximum capacity of the electron transport system (ETS)

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Blue Book chapter 1: Advanced questions

1 Given the formula for protonmotive force (pmF) as Δp = Δψ - 2.303 (RT/F) (ΔpH), where Δψ is the mitochondrial membrane potential, R is the gas constant, T is temperature in Kelvin, F is Faraday's constant, and ΔpH is the pH gradient across the mitochondrial membrane. If Δψ = 150 mV, T = 310 K, and ΔpH = 1, calculate the pmF in millivolts (mV). Assume R = 8.314 J/mol·K and F = 96485 C/mol.

	Approximately 170 mV
	Approximately 220 mV
	Approximately 130 mV
	The pmF cannot be calculated without additional data

2 The P/O ratio is an indicator of the efficiency of ATP synthesis relative to oxygen consumption. If 10 moles of ATP are produced for every 5 moles of oxygen consumed, what is the P/O ratio? What does this imply about the mitochondrial oxidative phosphorylation efficiency?

	P/O = 0.5; indicates a low efficiency of oxidative phosphorylation
	P/O = 2; indicates a high efficiency of oxidative phosphorylation
	P/O = 1; indicates a moderate efficiency of oxidative phosphorylation
	The P/O ratio is irrelevant to oxidative phosphorylation efficiency

3 Assuming the standard reduction potential (E°') for NADH → NAD+ is -0.320 V and for O₂ → H₂O is +0.815 V, calculate the ΔE°' for the electron transport from NADH to O₂. What does ΔE°' indicate about the potential energy available for ATP synthesis?

	ΔE°' = 0.495 V; indicates a moderate potential energy available for ATP synthesis
	ΔE°' = 1.135 V; indicates a high potential energy available for ATP synthesis

4 If the inner mitochondrial membrane has a surface area of 5.0 × 10⁶ μm² per mg of protein and each Complex I can pump 4 protons across the membrane, how many protons are pumped per second assuming a turnover number of 100 x s^-1 for Complex I?

	Calculation cannot be completed without the number of Complex I per μm^2
	2.0 × 109 protons x s-1
	5.0 × 109 protons x s-1
	2.0 × 109 protons x s-1

5 Using the Gibbs free energy equation ΔG = ΔG°' + RT ln(Q), where ΔG°' is the standard free energy change, R is the gas constant, T is the temperature in Kelvin, and Q is the reaction quotient. Calculate the ΔG for ATP synthesis if ΔG°' = -30.5 kJ/mol, T = 310 K, and the ATP/ADP ratio (Q) is 10. Assume R = 8.314 J/(mol·K).

	Additional information is needed to calculate ΔG
	-35.2 kJ/mol
	-45.6 kJ/mol
	-40.1 kJ/mol

6 The efficiency of mitochondrial oxidative phosphorylation can be described by the equation η = (ΔG_ATP/ΔG_O2) × 100%, where ΔG_ATP is the free energy change for ATP synthesis, and ΔG_O2 is the free energy change for oxygen reduction. If ΔG_ATP = -50 kJ/mol and ΔG_O2 = -200 kJ/mol, what is the efficiency (η) of oxidative phosphorylation?

	100 %
	25 %
	50 %
	75 %

7 Consider a mitochondrial uncoupling scenario where the membrane potential (Δψ) is decreased by 50% without altering the proton gradient (ΔpH). Using the Nernst equation for protons, E = (RT/zF)ln([H+]out/[H+]in), predict how this change affects the pmF. Assume R, T, F, and z values remain constant.

	Cannot predict without specific [H+]out/[H+]in values
	pmF remains unchanged because ΔpH is constant
	pmF decreases by 50 %
	pmF decreases, but not by 50 %

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Chapter 1.2 specific questions

1 Which mitochondrial preparation technique is most suitable for studying the effects of specific drugs on ATP production?

	Selectively permeabilized cells
	Whole-cell lysates
	Tissue homogenates
	Isolated mitochondrial fractions

2 In the context of mitochondrial diseases, why is it crucial to maintain the integrity of mitochondrial membranes during preparation?

	To prevent the release of mitochondrial DNA into the preparation medium
	To ensure the mitochondria can be visually distinguished under a microscope
	To enhance the structural appearance of mitochondria for photography
	To preserve the conditions necessary for accurate functional assays, such as measuring membrane potential

3 Match the mitochondrial preparation with its primary research application. Select the best match for "isolated mitochondrial fractions."

	Bioenergetic studies focusing on specific pathways
	General screenings for mitochondrial content
	Observations of mitochondrial behavior in living cells
	Structural analysis of mitochondrial networks

4 Considering the role of mitochondria in apoptosis, which aspect of mitochondrial preparations is crucial for studying their involvement in cell death mechanisms?

	The coloration of mitochondria for easier identification
	Maintaining the outer membrane's permeability to cytochrome c
	The size comparison between healthy and apoptotic mitochondria
	The ability to replicate mitochondrial DNA in vitro

5 Which statement best reflects the importance of studying mitochondrial bioenergetics in the context of metabolic diseases?

	It allows for the identification of new mitochondrial shapes
	It primarily aids in the classification of mitochondrial sizes
	The research is only relevant for academic purposes, not clinical applications
	Understanding mitochondrial function can lead to targeted therapies for diseases like diabetes

6 In the process of selectively permeabilizing cells for mitochondrial studies, what is the main goal?

	To allow specific molecules to access mitochondria while preserving overall cellular and mitochondrial structure
	To completely remove the cell nucleus
	To make mitochondria visible without staining
	To isolate mitochondria for genetic engineering purposes

7 How does the concept of "bioblasts" relate to modern mitochondrial research?

	It is a deprecated term with no relevance to current studies
	It highlights the historical view of mitochondria as autonomous entities
	It underscores the independence of mitochondria from cellular influence
	It emphasizes the integrated role of mitochondria within cellular bioenergetics

8 What advantage does using tissue homogenates offer in mitochondrial bioenergetic studies?

	They allow for the direct manipulation of mitochondrial DNA.
	They provide a means to study mitochondrial function in a context that includes interactions with other cell types and structures
	They simplify the study of mitochondria by removing all non-mitochondrial elements.
	They are used exclusively for determining the mitochondrial protein composition.

9 In mitochondrial preparations, why is the assessment of ATP synthesis capacity critical for understanding diseases like Parkinson's and Alzheimer's?

	Impaired ATP synthesis is a hallmark of many neurodegenerative conditions, affecting neuronal survival and function
	It helps in categorizing the diseases based on mitochondrial size.
	ATP synthesis capacity directly correlates with the severity of neurodegenerative diseases.
	It can reveal the evolutionary origins of these diseases.

10 Reflecting on the chapter's discussion, how do advancements in mitochondrial isolation techniques enhance our ability to treat metabolic disorders?

	By allowing for detailed study of mitochondrial function, leading to targeted therapeutic approaches
	They have no impact on treatment but offer insights into mitochondrial communication with extraterrestrial life
	Through the ability to transplant isolated mitochondria into patients
	By providing purely aesthetic insights into mitochondrial shape and structure

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