scholarly journals Human Platelet Mitochondrial Function Reflects Systemic Mitochondrial Alterations: A Protocol for Application in Field Studies

Cells ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 2088
Author(s):  
Florian Hoppel ◽  
Luiz Felipe Garcia-Souza ◽  
Wilhelm Kantner-Rumplmair ◽  
Martin Burtscher ◽  
Erich Gnaiger ◽  
...  
Keyword(s):  
Mitochondrial Function ◽  
Respiratory Control ◽  
Field Studies ◽  
Human Platelets ◽  
Chamber Volume ◽  
Mitochondrial Alterations ◽  
Human Blood Cells ◽  
Reliable Parameter ◽  
Systemic Stress ◽  
Control State

Human blood cells may offer a minimally invasive strategy to study systemic alterations of mitochondrial function. Here we tested the reliability of a protocol designed to study mitochondrial respiratory control in human platelets (PLTs) in field studies, using high-resolution respirometry (HRR). Several factors may trigger PLT aggregation during the assay, altering the homogeneity of the cell suspension and distorting the number of cells added to the two chambers (A, B) of the Oroboros Oxygraph-2k (O2k). Thus, inter-chamber variability (∆ab) was calculated by normalizing oxygen consumption to chamber volume (JO2) or to a specific respiratory control state (flux control ratio, FCR) as a reliable parameter of experimental quality. The method’s reliability was tested by comparing the ∆ab of laboratory-performed experiments (LAB, N = 9) to those of an ultramarathon field study (three sampling time-points: before competition (PRE, N = 7), immediately after (POST, N = 10) and 24 h after competition (REC; N = 10)). Our results show that ∆ab JO2 changed PRE-POST, but also for LAB-POST and LAB-REC, while all ∆ab FCR remained unchanged. Thus, we conclude that our method is reliable for assessing PLT mitochondrial function in LAB and field studies and after systemic stress conditions.

scholarly journals Effect of hyperbaric oxygen therapy on liver function during intermittent ischemia

2013 ◽  
Vol 28 (suppl 1) ◽  
pp. 61-65 ◽  
Author(s):  
Leticia Botigeli Baldim ◽  
Ricardo Nejo Jr ◽  
Maria Eliza Jordani Souza ◽  
Maria Cecília Jordani Gomes ◽  
Maria Aparecida Neves Cardoso Picinato ◽  
...  
Keyword(s):  
Liver Function ◽  
Hyperbaric Oxygen ◽  
Mitochondrial Function ◽  
Hyperbaric Oxygen Therapy ◽  
Aspartate Aminotransferase ◽  
Alanine Aminotransferase ◽  
Oxygen Therapy ◽  
Respiratory Control ◽  
Respiratory Control Ratio ◽  
Hepatic Pedicle

PURPOSE: To analyze the effects of hyperbaric oxygen therapy on liver function in rats previously subjected to ischemia and reperfusion. METHODS: A randomly distribution of 23 Wistar rats was conducted into three groups: SHAM, animals subjected to surgical stress without restricting blood flow by clamping the hepatic pedicle, IR, rats underwent hepatic vascular occlusion intermittently for two complete cycles of 15 minutes of ischemia followed by 5 min of reperfusion, IR / HBO, rats underwent hepatic pedicle clamping and thereafter exposed to hyperbaric oxygen pressure of 2 absolute atmospheres for 60 minutes. We evaluated liver function through mitochondrial function, determined by the stages 3 and 4 of respiration, respiratory control ratio (RCR) and mitochondrial permeability transition (Swelling). Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were also quantified . We analyzed the results using the Mann-Whitney test and were considered significant all results with p <0.05. RESULTS: There were significant differences between the results of stage 3 in SHAM vs IR group ; of the stage 4 in the groups IR vs SHAM and SHAM vs IR /HBO; of the Respiratory Control Ratio (RCR) in the group IR vs IR / HBO ; of alanine aminotransferase in the groups IR vs SHAM , SHAM vs IR/HBO and IR vs IR / HBO; aspartate aminotransferase in the groups SHAM vs IR and SHAM vs IR / HBO. CONCLUSION: The whole analysis of the mitochondiral function indicators permits us to conclude that the hyperbaric oxygen therapy acted as a protective agent of the mitochondrial function, minimizing the ischemia-reperfusion injury of the hepatic parenchyma.

Role of oxidative stress in alterations of mitochondrial function in ischemic-reperfused hearts

2007 ◽  
Vol 292 (4) ◽  
pp. H1986-H1994 ◽  
Author(s):  
Zhanna Makazan ◽  
Harjot K. Saini ◽  
Naranjan S. Dhalla
Keyword(s):  
Oxidative Stress ◽  
Oxidative Phosphorylation ◽  
Mitochondrial Function ◽  
Mitochondrial Respiration ◽  
Diastolic Pressure ◽  
Ischemia Reperfusion ◽  
Respiratory Control ◽  
Left Ventricular ◽  
Cardiac Performance ◽  
Developed Pressure

To study the mechanisms of mitochondrial dysfunction due to ischemia-reperfusion (I/R) injury, rat hearts were subjected to 20 or 30 min of global ischemia followed by 30 min of reperfusion. After recording both left ventricular developed pressure (LVDP) and end-diastolic pressure (LVEDP) to monitor the status of cardiac performance, mitochondria from these hearts were isolated to determine respiratory and oxidative phosphorylation activities. Although hearts subjected to 20 min of ischemia failed to generate LVDP and showed a marked increase in LVEDP, no changes in mitochondrial respiration and phosphorylation were observed. Reperfusion of 20-min ischemic hearts depressed mitochondrial function significantly but recovered LVDP completely and lowered the elevated LVEDP. On the other hand, depressed LVDP and elevated LVEDP in 30-min ischemic hearts were associated with depressions in both mitochondrial respiration and oxidative phosphorylation. Reperfusion of 30-min ischemic hearts elevated LVEDP, attenuated LVDP, and decreased mitochondrial state 3 and uncoupled respiration, respiratory control index, ADP-to-O ratio, as well as oxidative phosphorylation rate. Alterations of cardiac performance and mitochondrial function in I/R hearts were attenuated or prevented by pretreatment with oxyradical scavenging mixture (superoxide dismutase and catalase) or antioxidants [ N-acetyl-l-cysteine or N-(2-mercaptopropionyl)-glycine]. Furthermore, alterations in cardiac performance and mitochondrial function due to I/R were simulated by an oxyradical-generating system (xanthine plus xanthine oxidase) and an oxidant (H2O2) either upon perfusing the heart or upon incubation with mitochondria. These results support the view that oxidative stress plays an important role in inducing changes in cardiac performance and mitochondrial function due to I/R.

scholarly journals Behavior of cholinesterase and liver mitochondrial function in dogs submitted to normothermic ischemia and reperfusion

2003 ◽  
Vol 18 (suppl 5) ◽  
pp. 38-41
Author(s):  
Luis Pinto Fernandes ◽  
Ajith Kumar Sankarankutty ◽  
Eduardo Garcia Pacheco ◽  
Sérgio Centurion ◽  
Maria Cecília Jordani ◽  
...  
Keyword(s):  
Inferior Vena Cava ◽  
Portal Vein ◽  
Hepatic Artery ◽  
Mitochondrial Function ◽  
Inferior Vena ◽  
Vena Cava ◽  
Respiratory Control ◽  
Ischemic Injury ◽  
Sensitive Indicator ◽  
Ischemia And Reperfusion

PURPOSE: The plasmatic activity of the cholinesterase (CHE) and the liver mitochondrial function, expressed by the ratio of respiratory control (RCR), were studied during normothermic ischemia. METHODS: Sixteen adult mongrels, eight females and eight males were submitted to ischemia by clamping of the hepatic artery, portal vein and infrahepatic inferior vena cava, infra-hepatic, for two h, follwed by reperfusion for 1 h. The CHE and the mitochondrial function were evaluated at 60 and 120 min. of ischemia and at 15 and 60 minutes of reperfusion. RESULTS: The CHE decreased, significantly, during ischemia and in reperfusion. The RCR was decreased at 120 min. of ischemia, returning to the initial values on reperfusion. CONCLUSION: In this study, the CHE was a sensitive indicator of ischemic injury , suggesting irreversibility of ischemia injury. The RCR, by other side, showed a greater sensibility than the CHE in detection sense, during the studied period, the reversibility of the hepatic ischemic injury.

Drug Dependent Immune Destruction of Human Platelets in the NOD/SCID Mouse.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 3419-3419
Author(s):  
Daniel W. Bougie ◽  
Brian Boylan ◽  
Dhirendra Nayak ◽  
Peter J. Newman ◽  
Richard H. Aster
Keyword(s):  
Human Blood ◽  
Blood Cells ◽  
Scid Mouse ◽  
Human Platelets ◽  
Human Igg ◽  
Human Blood Cells ◽  
Rapid Clearance ◽  
Human Igg1 ◽  
Immune Destruction ◽  
Drug Dependent

Abstract Animal models have been of limited value for studying survival and immune destruction of human blood cells because of xenoantibodies that mediate rapid elimination of the transfused cells. The NOD/SCID mouse is a genetic variant that lacks immunoglobulins, including xenoantibodies, and may therefore be useful for studying survival and immune destruction of human blood cells (Newman, PJ et al. J Thromb Haemost.2007; 5 Suppl 1:305–9). Human platelets, infused into the retroorbital plexus of NOD/SCID mice survive about three days, nearly as long as mouse platelets themselves. Injection of IgG from a patient with quinine-induced immune thrombocytopenia (TP) and a newly developed quinine-dependent murine monoclonal antibody (mAb) were without effect on platelet survival. However, both antibodies caused rapid platelet clearance after IP injection of quinine. 500 μg, but not 50 μg, of IgG from serum containing an anti-HPA-1a (PlA1) antibody also caused rapid clearance of HPA-1a positive platelets. These findings indicate that the NOD/SCID mouse can be valuable for the study of immune clearance of human platelets and perhaps other unresolved problems in transfusion medicine. However, it is not known whether mice recognize cells opsonized with human IgG or mouse IgG equally well. To examine this, we compared the survival of human platelets sensitized with 1) mAb 7E3 (IgG1) specific for an epitope in the beta A domain of GPIIIa or 2) c7E3 (from Centocor Inc), which recognizes the same epitope but possesses a human IgG1 Fc. In titration studies, we found that about four times as much c7E3 as 7E3 is needed to produce equivalent clearance of human platelets, indicating that mouse IgG is a more efficient opsonin than human IgG. However, this difference should not limit use of the model for studies of antibody-mediated blood cell destruction, since mg quantities of IgG can easily be isolated from small amounts (0.1– 0.2 ml) of human serum.

Measurement of Mitochondrial Function in Human Platelets

2011 ◽  
Vol 51 ◽  
pp. S37
Author(s):  
Nayra Cardenes ◽  
Lisa Geary ◽  
Shilpa Jain ◽  
Catherine Corey ◽  
Christelle Kamga ◽  
...  
Keyword(s):  
Mitochondrial Function ◽  
Human Platelets

Exercise training in normobaric hypoxia in endurance runners. II. Improvement of mitochondrial properties in skeletal muscle

2006 ◽  
Vol 100 (4) ◽  
pp. 1249-1257 ◽  
Author(s):  
Elodie Ponsot ◽  
Stéphane P. Dufour ◽  
Joffrey Zoll ◽  
Stéphane Doutrelau ◽  
Benoit N'Guessan ◽  
...  
Keyword(s):  
Mitochondrial Function ◽  
Ventilatory Threshold ◽  
Vastus Lateralis ◽  
Respiratory Control ◽  
Endurance Performance ◽  
Oxidative Capacity ◽  
Time To Exhaustion ◽  
Intermittent Hypoxic Training ◽  
Minimal Speed ◽  
Before And After

This study investigates whether adaptations of mitochondrial function accompany the improvement of endurance performance capacity observed in well-trained athletes after an intermittent hypoxic training program. Fifteen endurance-trained athletes performed two weekly training sessions on treadmill at the velocity associated with the second ventilatory threshold (VT2) with inspired O2 fraction = 14.5% [hypoxic group (Hyp), n = 8] or with inspired O2 fraction = 21% [normoxic group (Nor), n = 7], integrated into their usual training, for 6 wk. Before and after training, oxygen uptake (V̇o2) and speed at VT2, maximal V̇o2 (V̇o2 max), and time to exhaustion at velocity of V̇o2 max (minimal speed associated with V̇o2 max) were measured, and muscle biopsies of vastus lateralis were harvested. Muscle oxidative capacities and sensitivity of mitochondrial respiration to ADP ( Km) were evaluated on permeabilized muscle fibers. Time to exhaustion, V̇o2 at VT2, and V̇o2 max were significantly improved in Hyp (+42, +8, and +5%, respectively) but not in Nor. No increase in muscle oxidative capacity was obtained with either training protocol. However, mitochondrial regulation shifted to a more oxidative profile in Hyp only as shown by the increased Km for ADP (Nor: before 476 ± 63, after 524 ± 62 μM, not significant; Hyp: before 441 ± 59, after 694 ± 51 μM, P < 0.05). Thus including hypoxia sessions into the usual training of athletes qualitatively ameliorates mitochondrial function by increasing the respiratory control by creatine, providing a tighter integration between ATP demand and supply.

scholarly journals Mitochondrial Dysfunction in Cancer Cachexia: Impact on Muscle Health and Regeneration

Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 3150
Author(s):  
Marc Beltrà ◽  
Fabrizio Pin ◽  
Riccardo Ballarò ◽  
Paola Costelli ◽  
Fabio Penna
Keyword(s):  
Skeletal Muscle ◽  
Mitochondrial Dysfunction ◽  
Mitochondrial Function ◽  
Cancer Cachexia ◽  
Compensatory Mechanism ◽  
Mitochondrial Alterations ◽  
The One ◽  
Energy Failure

Cancer cachexia is a frequently neglected debilitating syndrome that, beyond representing a primary cause of death and cancer therapy failure, negatively impacts on patients’ quality of life. Given the complexity of its multisystemic pathogenesis, affecting several organs beyond the skeletal muscle, defining an effective therapeutic approach has failed so far. Revamped attention of the scientific community working on cancer cachexia has focused on mitochondrial alterations occurring in the skeletal muscle as potential triggers of the complex metabolic derangements, eventually leading to hypercatabolism and tissue wasting. Mitochondrial dysfunction may be simplistically viewed as a cause of energy failure, thus inducing protein catabolism as a compensatory mechanism; however, other peculiar cachexia features may depend on mitochondria. On the one side, chemotherapy also impacts on muscle mitochondrial function while, on the other side, muscle-impaired regeneration may result from insufficient energy production from damaged mitochondria. Boosting mitochondrial function could thus improve the energetic status and chemotherapy tolerance, and relieve the myogenic process in cancer cachexia. In the present work, a focused review of the available literature on mitochondrial dysfunction in cancer cachexia is presented along with preliminary data dissecting the potential role of stimulating mitochondrial biogenesis via PGC-1α overexpression in distinct aspects of cancer-induced muscle wasting.

scholarly journals Effect of laser on the remnant liver after the first 24 hours following 70% hepatectomy in rats

2011 ◽  
Vol 26 (6) ◽  
pp. 470-474 ◽  
Author(s):  
Ailton Jose Barbosa ◽  
Ana Carolina Delazia Albuquerque Santana ◽  
Tiago Castro e Silva ◽  
Cristina Kurachi ◽  
Natalia Inada ◽  
...  
Keyword(s):  
Laser Radiation ◽  
Mitochondrial Function ◽  
Laser Light ◽  
Substantial Reduction ◽  
Respiratory Control ◽  
Control Group ◽  
Remnant Liver ◽  
Concomitant Increase ◽  
Group 4 ◽  
Male Wistar Rats

PURPOSE: To evaluate the mitochondrial function of the remnant liver (RL) in the early phase of liver regeneration in rats after 70% partial hepatectomy (PH). METHODS: Sixty male Wistar rats (200-250g) submitted to 70% PH were divided into five groups according to the time of euthanasia and application or not of laser light: C = Control, time zero; 2 minutes, 4, 6 and 24 hours after PH. The dose of laser radiation was 22.5 J/cm², wavelength of 660 nm (visible/red), in the remnant liver. We studied the respiration activated by ADP (state 3), basal mitochondrial respiration (state 4), respiratory control ratio (RCR) and mitochondrial membrane potential (MMP). RESULTS: The mitochondrial function of RL changed at 4 and 6 hours after PH, with a significant increase in state 3 and a concomitant increase in state 4 and with maintenance of RCR. MMP differed significantly between the groups biostimulated with laser radiation and the control group 4 hours after HP, with a substantial reduction in the non-laser groups. CONCLUSION: The laser light at the dose used in this study did not induce additional damage to the RL and seems to have delayed the hepatocellular metabolic overload of the remnant liver.

scholarly journals Calorie Restriction Maintains Mitochondrial Function and Redox Balance Avoiding Lipidomic Reprogramming during Isoproterenol-Induced Cardiac Hypertrophy

2021 ◽  
Author(s):  
Cícera Edna Barbosa David ◽  
Aline Maria Brito Lucas ◽  
Pedro Lourenzo Oliveira Cunha ◽  
Yuana Ivia Ponte Viana ◽  
Marcos Yukio Yoshinaga ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Cardiac Hypertrophy ◽  
Calorie Restriction ◽  
Mitochondrial Function ◽  
Cardiac Tissue ◽  
Unsaturated Fatty Acids ◽  
Saturated Fatty Acids ◽  
Respiratory Control ◽  
H2o2 Production

Cardiac hypertrophy induces a metabolic shift, leading to a preferential consumption of glucose (over fatty acids) to support the high energetic demand. Typically, health cardiac tissue utilizes more fat than any other organ. Calorie restriction is a dietary procedure that induces health benefits and lifespan extension in many organisms. Given the beneficial effects of calorie restriction and the metabolic dysregulation seen during cardiac hypertrophy, we hypothesized that calorie restriction prevents cardiac hypertrophy, lipid, mitochondrial, and redox dysregulations. Strikingly, calorie restriction reversed isoproterenol-induced cardiac hypertrophy, lowered succinate driven mitochondrial H2O2 production, improved mitochondrial function (indicated as a higher Respiratory Control Ratio – RCR) and avoided mitochondrial superoxide dismutase (MnSOD) and glutathione peroxidase (GPX) repression. To gain insight into how calorie restriction could interfere with the metabolic changes induced by cardiac hypertrophy, we performed lipidomic profiling. Calorie restriction protected against the consumption of several triglycerides (TG) linked to unsaturated fatty acids, and the accumulation of TGs containing saturated fatty acids observed in hypertrophic samples. Cardiac hypertrophy induced an increase in ceramides, phosphoethanolamines and acylcarnitines (12:0, 14:0, 16:0 and 18:0) that were also reversed by calorie restriction. Altogether, our data demonstrate that hypertrophy changes the cardiac lipidome, causes mitochondrial disturbances and oxidative stress. All these changes are prevented by calorie restriction intervention in vivo. This study uncovers calorie restriction as a resource protect cardiac tissue and prevent cardiac hypertrophy-induced lipidomic remodeling.

Assessment of Mitochondrial Respiration in Human Platelets

2017 ◽  
Vol 68 (4) ◽  
pp. 768-771
Author(s):  
Alexandra Petrus ◽  
Corina Ratiu ◽  
Lavinia Noveanu ◽  
Rodica Lighezan ◽  
Mariana Rosca ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Platelet Rich Plasma ◽  
Coupling Efficiency ◽  
Respiratory Control ◽  
Human Platelets ◽  
Mitochondrial Oxygen Consumption ◽  
Titration Protocol ◽  
Healthy Females

It has been long recognized that the impairment of platelet mitochondrial function occurs in a broad spectrum of diseases. Accordingly, the assessment of platelet respiratory dys/function has emerged as a putative approach allowing the characterization of the early impairment of human bioenergetic profile in several chronic pathologies. The aim of this study was to standardize the methodology for platelet isolation from peripheral blood and the measurement of mitochondrial oxygen consumption by means of high-resolution respirometry, respectively. The platelet isolation protocol consisted of two consecutive centrifugations of the whole blood collected from adult healthy females (n = 10) yielding a platelet-rich plasma sample. Respiration was measured at 370C using the Oxygraph-2k (Oroboros Instruments, Austria) according to a classic substrate-uncoupler-inhibitor-titration protocol. Platelets permeabilized with digitonin were allowed to respire in the presence of complex I (glutamate and malate) and complex II (succinate) substrates. We obtained a respiratory control ratio of 2.77 � 3.65 that indicates an accurate coupling efficiency of oxidative phosphorylation. The in vitro measurement of platelet respiration is a reliable method to evaluate the bioenergetic profile in humans. The standardized technique will be further used to assess the occurrence of mitochondrial dysfunction in peripheral blood in the setting of various chronic non-communicable diseases.

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