Oroboros Ecosystem Agenda 2024 Archive

Oroboros Ecosystem Agenda 2024 Archive

Mitochondria and Cell Research - the Oroboros Ecosystem

Last week

2024 Week 16

Apr-15 Mo

Do you observe either a zero or low raw O₂ signal when starting a measurement?

Perform a test to evaluate whether this is related to an improperly positioned chamber or a faulty connection between the OroboPOS and the OroboPOS-Connector.
- »Oxygen signal«
- communicated by »Grings Mateus« and »Schmitt Sabine«

Apr-16 Tu

Apr-17 We

Apr-18 Th

How to store the POS overnight and up to several months?

With the POS still in place, fill the O2k-chamber and receptacle of the loosely inserted stopper with 70 % EtOH, add the Cover-slip and switch off the O2K.
- »MiPNet19.03 O2k-cleaning and ISS«
- communicated by »Leo Elettra« and »Grings Mateus«

Apr-19 Fr

Apr 2024

2024 Week 14

Apr-01 Mo April - Parkinson's Disease Awareness Month

"Although the chain of events leading to this respiration deficit is currently not understood, the findings constitute a robust assay for the progressive pathology induced by PINK1 deficiency."

Gispert S, Ricciardi F, Kurz A, Azizov M, Hoepken HH, Becker D, Voos W, Leuner K, Müller WE, Kudin AP, Kunz WS, Zimmermann A, Roeper J, Wenzel D, Jendrach M, García-Arencíbia M, Fernández-Ruiz J, Huber L, Rohrer H, Barrera M, Reichert AS, Rüb U, Chen A, Nussbaum RL, Auburger G (2009) Parkinson phenotype in aged PINK1-deficient mice is accompanied by progressive mitochondrial dysfunction in absence of neurodegeneration. »Bioblast link«

from the O2k-Network »US CA San Francisco Nussbaum RL« and »DE Frankfurt Eckert GP«

Apr-02 Tu

Apr-03 We

Apr-04 Th

Why is it important to check the H₂O₂ sensitivity when conducting an Amplex UltraRed (AmR) assay?

The sensitivity is influenced by experimental time, medium composition, added chemicals and sample, requiring multiple H₂O₂ calibrations throughout the measurement. After the initial AmR calibration, ensure that the sensitivity is within the recommended range.

- »MiPNet24.10 H2O2 flux analysis«, »Amplex UltraRed«
- communicated by »Grings Mateus« and »Schmitt Sabine«

Apr-05 Fr

2024 Week 15

Apr-08 Mo

16:30 Erich Gnaiger presenting a MOSBRI Consensus Meeting satellite seminar at O2k-Network Lab IT Rome Giuffre A - Unlocking cell metabolism: Exploring the power of high-resolution respirometry with the NextGen-O2k

Apr-09 Tu

10:00 Erich Gnaiger presenting at MOSBRI Consensus Meeting 2024 Rome IT - Quality-controlled high-resolution respirometry: bacterial and clinical applications

Apr-10 We

Is fatty acid oxidation (FAO) considered as CI- or CII-driven respiration?

Neither. There is simultaneous electron transfer from FAO into the Q-junction through the electron transferring flavoprotein dehydrogenase complex (CETFDH) and Complex I.
- »Fatty acid oxidation«, »Gnaiger 2020 BEC MitoPathways«, »Gnaiger 2024 J Biol Chem«
- communicated by »Timon-Gomez Alba« and »Willis Jaime R«

Apr-11 Th

World Parkinson's Disease Day

"The markers of mitochondrial biogenesis were consistently affected in PD [Parkinson’s disease] patients group."

Kamienieva I, Charzyńska A, Duszyński J, Malińska D, Szczepanowska J (2023) In search for mitochondrial biomarkers of Parkinson's disease: Findings in parkin-mutant human fibroblasts. »Bioblast link«

from the O2k-Network »PL Warsaw Szewczyk A«

Apr-12 Fr

Mar 2024

2024 Week 10

Mar-04 Mo

Do you observe an unstable O₂ slope negative following TIP2k titrations during an instrumental O₂ background test?

Incorrect placement of TIP2k Filter Papers can lead to liquid accumulation and convective oxygen backdiffusion. Check for proper placement by confirming they are fully in contact with the stopper receptacle.
- »Instrumental O₂ background«, »Unstable O₂ slope negative«
- communicated by »Grings Mateus« and »Baglivo Eleonora«

Mar-05 Tu

Mar-06 We

The tale of two “best-friends,” proteins BID & MTCH2, pivotal regulators of the apoptosis and metabolism programs at the mitochondria.

Gross A, (2024) A BID-MTCH2 love story: Energizing mitochondria until apoptosis do them part? Bioenerg Commun 2024.1. https://doi.org/10.26124/bec:2024-0001

Mar-07 Th

Which medium can be used for the instrumental background test?

When using dithionite, the instrumental background test should be performed in mitochondrial respiration medium MiR05, MiR05-Kit or MiR06; or alternatively, in a strong alkaline phosphate buffer (>100 mM; >pH 8), to avoid side reactions that might interfere with the test.
- »MiPNet14.06_Instrumental_O2_background«
- communicated by »Schmitt Sabine« and »Timon-Gomez Alba«

Mar-08 Fr

2024 Week 11

Mar-11 Mo

Why is it important to perform respirometry with permeabilized muscle fibers (pfi) under a high [O₂] regime?

Long diffusion distances in the pfi result in an O₂ diffusion limitation, which is overcome by increasing the O₂ partial pressure inside the chamber. Proper instrumental background correction is a must and O₂ dependence of ROS production should be considered.
- »Permeabilized muscle fibers«, »Oxygen flux - instrumental background«
- communicated by »Grings Mateus« and »Cardoso Luiza«

Mar-12 Tu

Mar-13 We

Rewarded Open Access - compensating authors

Together with the MiPsociety, BEC is raising funds to reward the first 10 papers accepted for publication in BEC in 2024: 600 EUR to attend EBEC2024 and 2-year MiPmembership. -»Support open access in science«
- »Submit a manuscript«

Mar-14 Th

Mitochondrial ultrastructure and respiration are affected during EDL [Extensor digitorum longus muscle] aging.

Fernando R, Shindyapina AV, Ost M, Santesmasses D, Hu Y, Tyshkovskiy A, Yim SH, Weiss J, Gladyshev VN, Grune T, Castro JP (2023) Downregulation of mitochondrial metabolism is a driver for fast skeletal muscle loss during mouse aging. »Bioblast link«

from the O2k-Network »DE Leipzip Ost M«

Mar-15 Fr

2024 Week 12

Mar-18 Mo

»EU-METAHEART MC2 Meeting 2024 Antalya TR« Mar 18-21

Mar-19 Tu

15:05 Erich Gnaiger presenting at EU-METAHEART MC2 Meeting 2024 Antalya TR - Functional hypoxia in cardiac mitochondria: effects on oxidative phosphorylation, mitochondrial membrane potential, redox state of coenzyme Q, and calcium uptake

Mar-20 We

What if Alert: Error in titrations! appears in your O2 analysis template file?

The O2 analysis template recognizes discrepancies between the protocol selected in the first tab and the imported marks. Check the titration sequence and mark names against the protocol copied from the DLD file.
- »O₂ flux analysis«
- communicated by »Timon-Gomez Alba« and »Leo Elettra«

Mar-21 Th

»Club17 2024 Innsbruck AT«

17:00 Verena Laner presenting at Club17 2024 Innsbruck AT - Functional Mitochondrial Diagnostics

How well do you handle your Hamilton syringes? What is a cleaning cycle?

"A syringe cleaning cycle consists of (1) filling an empty syringe with a washing solution while immersing [both the needle and] the base of the barrel into the washing solution, without fully removing the plunger, and (2) emptying the syringe into the waste beaker."
- »MiPNet19.14 SOP Hamilton microsyringes«
- communicated by »Baglivo Eleonora« and »Timon-Gomez Alba«

Mar-22 Fr

2024 Week 13

Mar-25 Mo

Living cells or intact cells?

Living cells are frequently labelled as intact cells in the sense of the total cell count, but intact may suggest dual meanings of viable or unaffected by a disease or mitochondrial injury.
- »Living cells«, »Gnaiger 2020 BEC Mitochondrial physiology«, »Gnaiger 2020 BEC MitoPathways«
- communicated by »Leo Elettra« and »Timon-Gomez Alba«

Mar-26 Tu

Mar-27 We

Mar-28 Th

"Introduction to the new models available with the NextGen was immensely helpful. This workshop has been critical for implementation of new techniques and advancement of my research."

Zunica Elizabeth, from the O2k-Network »US LA Baton Rouge Noland RC«
- »O2k-Feedback«

Mar-29 Fr

Feb 2024

2024 Week 06

Feb-05 Mo

"We demonstrated deficient platelet mitochondrial respiratory chain functions, oxidative phosphorylation (OXPHOS), and electron transfer (ET) capacity with complex I (CI)-linked substrates, and reduced [...] endogenous platelet coenzyme Q10 (CoQ10) concentration in UC [urothelial carcinoma] patients."

Palacka P, Gvozdjáková A, Rausová Z, Kucharská J, Slopovský J, Obertová J, Furka D, Furka S, Singh KK, Sumbalová Z (2021) Platelet mitochondrial bioenergetics reprogramming in patients with urothelial carcinoma. »Bioblast link«

from the O2k-Network »SK Bratislava Sumbalova Z«

Feb-06 Tu

Feb-07 We

Feb-08 Th

Can I use the same stock solutions for all reagents for different O2k-chamber volumes?

For some reagents, the concentration of the stock solution differs between the 2.0-mL and the 0.5-mL chamber (O2k-sV-Module) to facilitate a precise volume titration.
-»MiPNet09.12 O2k-Titrations«
- communicated by »Timon-Gomez Alba« and »Leo Elettra«

Feb-09 Fr

Feb-10 Sa

2024 Week 07

Feb-12 Mo

What is the O₂ solubility factor of the medium (F_M) and why is it important?

F_M expresses the effect of the salt concentration on oxygen solubility relative to pure water, and varies depending on the temperature and composition of the medium. The correct F_M needs to be entered in DatLab when performing an air calibration.
- »Oxygen solubility factor« , »Air calibration in DatLab«
- communicated by »Baglivo Eleonora« and »Timon-Gomez Alba«

Feb-13 Tu

Feb-14 We

Feb-15 Th - International Childhood Cancer Day

"Deleterious mtDNA mutations contribute significantly to the tumorigenesis of pediatric cancers"

Triska P, Kaneva K, Merkurjev D, Sohail N, Falk MJ, Triche TJ Jr, Biegel JA, Gai X (2019) Landscape of germline and somatic mitochondrial DNA mutations in pediatric malignancies. »Bioblast link«

from the O2k-Network »US_PA_Philadelphia_Falk_MJ«

Feb-16 Fr

2024 Week 08

Feb-19 Mo

“We just published this paper on SOMMA, possibly one of the largest aging studies involving permeabilized muscle energetics. Couldn't have done it without the training your teams provided.”

Kramer Philip A from the O2k-Network Lab »US NC Winston-Salem Kramer PA«
- »O2k-Feedback«

Feb-20 Tu

Feb-21 We

Feb-22 Th

What to do if the glass of the O2k-chamber starts to become opaque?

"Turbidity on the glass wall may be caused by precipitated protein. Remove the chambers from the O2k and with the O2k-stirrers removed, immerse in a beaker filled with HCl (10 M) overnight under a hood."
-»MiPNet19.03_O2k-cleaning_and_ISS«
- communicated by »Cardoso Luiza« and »Leo Elettra«

Feb-23 Fr

2024 Week 09

Feb-26 Mo

ATP synthase = phosphorylation system?

No, the phosphorylation system consists of adenine nucleotide translocase, phosphate carrier, and ATP synthase, which carry out the steps in the phosphorylation pathway.
- »Phosphorylation pathway«, »Mitochondrial Pathways: Chapter 2.2«
- communicated by »Schmitt Sabine« and »Timon-Gomez Alba«

Feb-27 Tu

Feb-28 We

Feb-29 Th

"Bed rest results in a decrease in skeletal muscle mitochondrial respiratory capacity, mainly as a result of an observed decrease in mitochondrial content"

Noone J, Damiot A, Kenny H, Chery I, Zahariev A, Normand S, Crampes F, de Glisezinski I, Rochfort KD, Laurens C, Bareille MP, Simon C, Bergouignan A, Blanc S, O'Gorman DJ (2023) The impact of 60 days of -6° head down tilt bed rest on mitochondrial content, respiration and regulators of mitochondrial dynamics. »Bioblast link«

from the O2k-Networks »US FL Orlando Goodpaster BH« , »FR Strasbourg Blanc S« , »IE Dublin O Gorman D«

Mar-01 Fr

Jan 2024

2024 Week 01

Jan-01 Mo

Jan-02 Tu

New year, new state? What is REN state?

REN state is now distinguished from ROX state: REN is measured in mitochondrial preparations without addition of substrates, whereas ROX is due to oxidative side reactions remaining after inhibition of the electron transfer pathway.
- »REN«, »ROX«
- communicated by »Timon-Gomez Alba« and »Leo Elettra«

Jan-03 We

Jan-04 Th

"Oxygen Consumption Rate (OCR) measurements of fresh and cryopreserved placental specimens are comparable whereas snap frozen procedure impairs mitochondrial activity"

Giovarelli M, Serati A, Zecchini S, Guelfi F, Clementi E, Mandò C (2023) Cryopreserved placental biopsies maintain mitochondrial activity for high-resolution respirometry. »Bioblast link«

from the O2k-Network »IT Milan Clementi E«

Jan-05 Fr

2024 Week 02

Jan-08 Mo

Do you need to export DatLab traces for your publication?

Save your graph as an editable picture using "Copy to clipboard" in DatLab 7.4 (.wmf) or "Save" in DatLab 8 (.svg).
- »Copy to clipboard in DatLab 7«, »Download DatLab 8«
- communicated by »Baglivo Eleonora« and »Grings Mateus«

Jan-09 Tu

Jan-10 We

»AICBC 2024 Navi Mumbai IN« Jan 10-12

Jan-11 Th

What are special features of the new SmartPOS (O2k-Series J and NextGen-O2k-Series XB)?

- "The electrical connections of the SmartPOS are protected thus there is no need to clean them."
- "The serial number of the SmartPOS is automatically recognized by the DatLab 8 software."

»MiPNet28.10 SmartPOS-service«
- communicated by »Grings Mateus« and »Baglivo Eleonora«

Jan-12 Fr

2024 Week 03

Jan-15 Mo

Why electron transfer pathway and not electron transfer chain?

The term “chain” suggests a linear sequence, however, the convergence of electron flow at the Q-junction indicates a higher degree of complexity, making “pathway” a more fitting descriptor.
- »Electron transfer pathway«, »Gnaiger 2020 BEC MitoPathways«
- communicated by »Leo Elettra« and »Timon-Gomez Alba«

Jan-16 Tu

Jan-17 We

Jan-18 Th

The study on rats by Jedlička et al indicates that "...the concept of general pattern of age-dependent decline in mitochondrial oxygen consumption across different organs and tissues could be misleading."

Jedlička J, Tůma Z, Razak K, Kunc R, Kala A, Proskauer Pena S, Lerchner T, Ježek K, Kuncová J (2022) Impact of aging on mitochondrial respiration in various organs. »Bioblast link«

from the O2k-Network »CZ_Pilsen_Kuncova_J«

Jan-19 Fr

2024 Week 04

Jan-22 Mo

“We are truly glad to have your continued support and partnership.”

Lu Amelia from the O2k-Network Lab »TW Taoyuan Sunpoint«
- »O2k-Feedback«

Jan-23 Tu

Jan-24 We

Jan-25 Th

How do I view and change the settings of a measurement/file in DatLab 8?

"O2k control (active) will always show the settings of the active recording [and allow changes], while O2k info (view) will show the settings of the recorded selected file."

-»MiPNet28.07 NextGen-O2k Series XB manual«; »MiPNet28.08 Oroboros O2k Series J manual«
- communicated by »Baglivo Eleonora« and »Grings Mateus«

Jan-26 Fr

 2024 Week 05

Jan-29 Mo

»E-mit Winter School 2024 Brixen IT« Jan 28 - Feb 02

Jan-30 Tu

16:30 Erich Gnaiger presenting at E-mit Winter School 2024 Brixen IT - Coupling control and pathway control in mitochondrial high-resolution respirometry

Jan-31 We

Can the S-pathway O₂ flux be derived by subtracting N-pathway O₂ flux from NS-pathway O₂ flux?

No, the sum of the O₂ fluxes measured in the presence of NADH-linked substrates alone and succinate as the single substrate is typically higher than using a combination of all these substrates. This effect is described as incomplete additivity:
- »Additive effect of convergent electron flow«
- communicated by »Cardoso Luiza« and »Grings Mateus«

Feb-01 Th

“Doing the entire hands-on module from start to finish was the best for really “cementing” theory and hands-on application together.”

Cosgrove Richard from Eli Lilly Research Labs, on what he liked best on IOC162
- »O2k-Feedback«

Feb-02 Fr