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Frontiers in Physiology 2023Mitochondrial dysfunction is linked to a variety of human diseases. Understanding the dynamic alterations in mitochondrial respiration at various stages of development...
Mitochondrial dysfunction is linked to a variety of human diseases. Understanding the dynamic alterations in mitochondrial respiration at various stages of development is important to our understanding of disease progression. Zebrafish provide a system for investigating mitochondrial function and alterations during different life stages. The purpose of this study was to investigate our ability to measure mitochondrial oxygen consumption rates in zebrafish embryos, larvae, and adults as an indicator of mitochondrial function. Basal respiration of entire zebrafish embryos (5 dpf), larvae (0.6-0.9 cm), young adults (3-month-old), and old adults (12-month-old) was measured using an Oroboros Oxygraph, with a stirrer speed of 26 rpm. For embryos and larvae, "leak" respiration (plus oligomycin), maximum respiration (plus uncoupler), non-mitochondrial respiration (plus inhibitors), and complex IV activity were also measured. To induce physical activity in adult fish, the stirrer speed was increased to 200 rpm. We demonstrate the ability to accurately measure respiration rates in zebrafish at various ages using the Oroboros Oxygraph. When comparing zebrafish embryos to larvae, embryos have a higher maximum respiration. Three-month-old zebrafish males have higher basal respiration than females, while 12-month-old zebrafish females exhibit greater rates of respiration than males and younger females. When the stirrer speed was increased, respiration rates decrease, but with differences depending on sex. This study demonstrates a simple and accessible method to assess zebrafish physiology by mitochondrial oxygen consumption measurements in an unmodified Oroboros Oxygraph. The method should facilitate studies to understand the intricate interplay between mitochondrial function, development, and aging.
PubMed: 37781232
DOI: 10.3389/fphys.2023.1272366 -
Scientific Reports Sep 2023Some of the prominent features of long-term memory formation include protein synthesis, gene expression, enhanced neurotransmitter release, increased excitability, and...
Some of the prominent features of long-term memory formation include protein synthesis, gene expression, enhanced neurotransmitter release, increased excitability, and formation of new synapses. As these processes are critically dependent on mitochondrial function, we hypothesized that increased mitochondrial respiration and dynamics would play a prominent role in memory formation. To address this possibility, we measured mitochondrial oxygen consumption (OCR) in hippocampal tissue punches from trained and untrained animals. Our results show that context fear training significantly increased basal, ATP synthesis-linked, and maximal OCR in the Shaffer collateral-CA1 synaptic region, but not in the CA1 cell body layer. These changes were recapitulated in synaptosomes isolated from the hippocampi of fear-trained animals. As dynamin-related protein 1 (Drp1) plays an important role in mitochondrial fission, we examined its role in the increased mitochondrial respiration observed after fear training. Drp1 inhibitors decreased the training-associated enhancement of OCR and impaired contextual fear memory, but did not alter the number of synaptosomes containing mitochondria. Taken together, our results show context fear training increases presynaptic mitochondria respiration, and that Drp-1 mediated enhanced energy production in CA1 pre-synaptic terminals is necessary for context fear memory that does not result from an increase in the number of synaptosomes containing mitochondria or an increase in mitochondrial mass within the synaptic layer.
Topics: Animals; Biological Transport; Oxygen Consumption; Synapses; Memory Disorders; Mitochondria
PubMed: 37660191
DOI: 10.1038/s41598-023-40877-0 -
Sensors (Basel, Switzerland) Nov 2023New and promising variables are being developed to analyze performance and fatigue in trail running, such as mechanical power, metabolic power, metabolic cost of...
New and promising variables are being developed to analyze performance and fatigue in trail running, such as mechanical power, metabolic power, metabolic cost of transport and mechanical efficiency. The aim of this study was to analyze the behavior of these variables during a real vertical kilometer field test. Fifteen trained trail runners, eleven men (from 22 to 38 years old) and four women (from 19 to 35 years old) performed a vertical kilometer with a length of 4.64 km and 835 m positive slope. During the entire race, the runners were equipped with portable gas analyzers (Cosmed K5) to assess their cardiorespiratory and metabolic responses breath by breath. Significant differences were found between top-level runners versus low-level runners in the mean values of the variables of mechanical power, metabolic power and velocity. A repeated-measures ANOVA showed significant differences between the sections, the incline and the interactions between all the analyzed variables, in addition to differences depending on the level of the runner. The variable of mechanical power can be statistically significantly predicted from metabolic power and vertical net metabolic COT. An algebraic expression was obtained to calculate the value of metabolic power. Integrating the variables of mechanical power, vertical velocity and metabolic power into phone apps and smartwatches is a new opportunity to improve performance monitoring in trail running.
Topics: Male; Humans; Female; Young Adult; Adult; Oxygen Consumption; Running; Energy Metabolism; Fatigue; Biomechanical Phenomena
PubMed: 38067721
DOI: 10.3390/s23239349 -
PloS One 2023Otopetrin 1 (OTOP1) is a proton channel that is highly expressed in brown adipose tissue. We examined the physiology of Otop1-/- mice, which lack functional OTOP1.
OBJECTIVE
Otopetrin 1 (OTOP1) is a proton channel that is highly expressed in brown adipose tissue. We examined the physiology of Otop1-/- mice, which lack functional OTOP1.
METHODS
Mice were studied by indirect calorimetry and telemetric ambulatory body temperature monitoring. Mitochondrial function was measured as oxygen consumption and extracellular acidification.
RESULTS
Otop1-/- mice had similar body temperatures as control mice at baseline and in response to cold and hot ambient temperatures. However, in response to fasting the Otop1-/- mice exhibited an exaggerated hypothermia and hypometabolism. Similarly, in ex vivo tests of Otop1-/- brown adipose tissue mitochondrial function, there was no change in baseline oxygen consumption, but the oxygen consumption was reduced after maximal uncoupling with FCCP and increased upon stimulation with the β3-adrenergic agonist CL316243. Mast cells also express Otop1, and Otop1-/- mice had intact, possibly greater hypothermia in response to mast cell activation by the adenosine A3 receptor agonist MRS5698. No increase in insulin resistance was observed in the Otop1-/- mice.
CONCLUSIONS
Loss of OTOP1 does not change basal function of brown adipose tissue but affects stimulated responses.
Topics: Animals; Mice; Adipose Tissue, Brown; Body Temperature; Body Temperature Regulation; Fasting; Hypothermia; Mice, Knockout
PubMed: 37812612
DOI: 10.1371/journal.pone.0292610 -
APL Bioengineering Sep 2023Accurately modeling oxygen transport and consumption is crucial to predict metabolic dynamics in cell cultures and optimize the design of tissue and organ models. We...
Accurately modeling oxygen transport and consumption is crucial to predict metabolic dynamics in cell cultures and optimize the design of tissue and organ models. We present a methodology to characterize the Michaelis-Menten oxygen consumption parameters , integrating novel experimental techniques and computational tools. The parameters were derived for hepatic cell cultures with different dimensionality (i.e., 2D and 3D) and with different surface and volumetric densities. To quantify cell packing regardless of the dimensionality of cultures, we devised an image-based metric, referred to as the proximity index. The Michaelis-Menten parameters were related to the proximity index through an uptake coefficient, analogous to a diffusion constant, enabling the quantitative analysis of oxygen dynamics across dimensions. Our results show that Michaelis-Menten parameters are not constant for a given cell type but change with dimensionality and cell density. The maximum consumption rate per cell decreases significantly with cell surface and volumetric density, while the Michaelis-Menten constant tends to increase. In addition, the dependency of the uptake coefficient on the proximity index suggests that the oxygen consumption rate of hepatic cells is superadaptive, as they modulate their oxygen utilization according to its local availability and to the proximity of other cells. We describe, for the first time, how cells consume oxygen as a function of cell proximity, through a quantitative index, which combines cell density and dimensionality. This study enhances our understanding of how cell-cell interaction affects oxygen dynamics and enables better prediction of aerobic metabolism in tissue models, improving their translational value.
PubMed: 37664826
DOI: 10.1063/5.0160422 -
Arquivos Brasileiros de Cardiologia 2024
Topics: Humans; Postural Balance; Time and Motion Studies; Heart Diseases; Oxygen Consumption
PubMed: 38451619
DOI: 10.36660/abc.20230832 -
Perfusion Jul 2023Isolated limb perfusion (ILP) is a regional surgical treatment for localized metastatic disease. High doses of chemotherapeutic agents are administered within an...
BACKGROUND
Isolated limb perfusion (ILP) is a regional surgical treatment for localized metastatic disease. High doses of chemotherapeutic agents are administered within an extracorporeal circulated isolated extremity, treating the metastasis, while systemic toxicity is avoided. To our knowledge, indexed oxygen supply/demand relationship during ILP has not previously been described. Our aim was to measure and describe oxygen metabolism, specifically oxygen delivery, consumption, and extraction, in an isolated leg/arm during ILP. Also investigate whether invasive oxygenation measurement during ILP correlates and can be used interchangeable with the non-invasive method, near infrared spectroscopy (NIRS).
METHODS
Data from 40 patients scheduled for ILP were included. At six time points blood samples were drawn during the procedure. DO2, VO2, and O2ER were calculated according to standard formulas. NIRS and hemodynamics were recorded every 10 min.
RESULTS
For all observations, the mean of DO2 was 190±59 ml/min/m2, VO2 was 35±8 ml/min/m2, and O2ER was 21±8%. VO2 was significantly higher in legs compared to arms (38±8 vs. 29±7 ml/min/m2, p=0.02). Repeated measures showed a significant decrease in DO2 in legs (209±65 to 180±66 ml/min/m2, p=<0.01) and in arms (252±72 to 150±57 ml/min/m2, p=<0.01). Significant increase in O2ER in arms was also found (p=0.03). Significant correlation was detected between NIRS and venous extremity oxygen saturation (SveO2) (rrm=0.568, p=<. 001, 95% CI 0.397-0.701). When comparing SveO2 and NIRS using a Bland-Altman analysis, the mean difference (bias) was 8.26±13.03 (p=<. 001) and the limit of agreement was - 17.28-33.09, with an error of 32.5%.
CONCLUSION
DO2 above 170 ml/min/m2 during ILP kept O2ER below 30% for all observations. NIRS correlates significant to SveO2; however, the two methods do not agree sufficiently to work interchangeable. Clinical Trial Registration URL: https://www.clinicaltrials.gov. Unique identifier: NCT04460053 and NCT03073304.
Topics: Humans; Extracorporeal Circulation; Extremities; Hemodynamics; Oxygen; Oxygen Consumption; Perfusion
PubMed: 35575302
DOI: 10.1177/02676591221093201 -
Frontiers in Oncology 2024Head and neck squamous cell carcinoma (HNSCC) patients suffer from frequent local recurrences that negatively impact on prognosis. Hence, distinguishing tumor and normal...
INTRODUCTION
Head and neck squamous cell carcinoma (HNSCC) patients suffer from frequent local recurrences that negatively impact on prognosis. Hence, distinguishing tumor and normal tissue is of clinical importance as it may improve the detection of residual tumor tissue in surgical resection margins and during imaging-based surgery planning. Differences in O consumption (OC) can be used to this aim, as they provide options for improved surgical, image-guided approaches.
METHODS
In the present study, the potential of a fluorescent sensor foil-based technology to quantify OC in HNSCC was evaluated in an 3D model and in situ in patients.
RESULTS
measurements of OC using hypopharyngeal and esophageal cell lines allowed a specific detection of tumor cell spheroids embedded together with cancer-associated fibroblasts in type I collagen extracellular matrix down to a diameter of 440 µm. Pre-surgery in situ measurements were conducted with a handheld recording device and sensor foils with an oxygen permeable membrane and immobilized O-reactive fluorescent dyes. Lateral tongue carcinoma and carcinoma of the floor of the mouth were chosen for analysis owing to their facilitated accessibility. OC was evaluated over a time span of 60 seconds and was significantly higher in tumor tissue compared to healthy mucosa in the vicinity of the tumor.
DISCUSSION
Hence, OC quantification using fluorescent sensor foil-based technology is a relevant parameter for the differentiation of tumor tissue of the head and neck region and may support surgery planning.
PubMed: 38390268
DOI: 10.3389/fonc.2024.1002798 -
Insects Jul 2023Hermetic storage methods are effective at protecting grain against insect pests. Biotic and abiotic factors influence oxygen depletion during hermetic storage. We...
Hermetic storage methods are effective at protecting grain against insect pests. Biotic and abiotic factors influence oxygen depletion during hermetic storage. We investigated the dual effects of temperature and initial pest infestation level on oxygen depletion during airtight storage. Glass jars filled with cowpea grain were infested (25 or 50 adult cowpea bruchids), then hermetically sealed and stored at 20, 30, or 40 °C for 30 days. Oxygen depletion, relative humidity, and temperature were monitored. Germination, grain moisture content, grain damage and weight loss, and adult emergence were assessed. Oxygen depletion varied by temperature and insect infestation level. However, 30 °C was the optimum temperature for oxygen depletion (reaching 5% or less in 10 days) regardless of insect infestation level. No changes were observed in germination and grain moisture content, minimal grain damage, or weight loss (<1%). Only at 20 °C were adult insects able to survive after 30 days and emerged 45 days post-treatment under normoxia. Therefore, hermetic storage containers should remain closed for more than 30 days to minimize re-infestation of grain in areas where average ambient temperatures rarely exceed 23 °C. Further research is needed to assess the effect of low temperatures on oxygen depletion and insect survival in hermetic storage beyond 30 days.
PubMed: 37504627
DOI: 10.3390/insects14070621 -
Biosensors Feb 2024Oxygen consumption has been used to evaluate various cellular activities. In addition, three-dimensional (3D) spheroids have been broadly exploited as advanced in vitro...
Oxygen consumption has been used to evaluate various cellular activities. In addition, three-dimensional (3D) spheroids have been broadly exploited as advanced in vitro cell models for various biomedical studies due to their capability of mimicking 3D in vivo microenvironments and cell arrangements. However, monitoring the oxygen consumption of live 3D spheroids poses challenges because existing invasive methods cause structural and cell damage. In contrast, optical methods using fluorescence labeling and microscopy are non-invasive, but they suffer from technical limitations like high cost, tedious procedures, and poor signal-to-noise ratios. To address these challenges, we developed a microfluidic platform for uniform-sized spheroid formation, handling, and culture. The platform is further integrated with widefield frequency domain fluorescence lifetime imaging microscopy (FD-FLIM) to efficiently characterize the lifetime of an oxygen-sensitive dye filling the platform for oxygen consumption characterization. In the experiments, osteosarcoma (MG-63) cells are exploited as the spheroid model and for the oxygen consumption analysis. The results demonstrate the functionality of the developed approach and show the accurate characterization of the oxygen consumption of the spheroids in response to drug treatments. The developed approach possesses great potential to advance spheroid metabolism studies with single-spheroid resolution and high sensitivity.
Topics: Microfluidics; Spheroids, Cellular; Microscopy, Fluorescence; Oxygen; Oxygen Consumption
PubMed: 38392015
DOI: 10.3390/bios14020096