A mitohormetic response to pro-oxidant exposure in the house mouse.

Mitochondria are hypothesized to display a biphasic response to reactive oxygen species (ROS) exposure. In this study, we evaluated the time course changes in mitochondrial performance and oxidative stress in house mice following X-irradiation. Forty-eight mice were equally divided among six groups, including a nonirradiated control and five experimental groups that varied in time between X-ray exposure and euthanasia (1 h and 1, 4, 7, and 10 days after X-irradiation). We measured parameters associated with mitochondrial respiratory function and ROS emission from isolated liver and skeletal muscle mitochondria and levels of oxidative damage and antioxidants in liver, skeletal muscle, and heart tissues. Mitochondrial function dropped initially after X-irradiation but recovered quickly and was elevated 10 days after the exposure. Hydrogen peroxide production, lipid peroxidation, and protein carbonylation showed inverse U-shaped curves, with levels returning to control or lower than control, 10 days after X-irradiation. Enzymatic antioxidants and markers for mitochondrial biogenesis exhibited a tissue-specific response after irradiation. These data provide the first chronological description of the mitohormetic response after a mild dose of irradiation and highlight the protective response that cells display to ROS exposure. This study also provides valuable information and application for future mitochondrial and oxidative stress studies in numerous physiological settings.

Photobiomodulation increases mitochondrial citrate synthase activity in rats submitted to aerobic training.

This study investigated the effects of photobiomodulation by low-laser laser therapy (LLLT) on the activities of citrate synthase (CS) and lactate dehydrogenase (LDH) and the anaerobic threshold (AT) in rats submitted to treadmill exercise. Fifty-four rats were allocated into four groups: rest control (RCG), rest laser (RLG), exercise control (ECG), and exercise laser (ELG). The infrared LLLT was applied daily on the quadriceps, gluteus maximum, soleus, and tibialis anterior muscles. Muscle samples (soleus, tibialis anterior, and cardiac muscles) were removed 48 h after the last exercise session for spectrophotometric analysis of the CS and LDH. The CS activity (µmol/protein) in ELG (16.02 and 0.49) was significantly greater (P < 0.05) than RCG (2.34 and 0.24), RLG (6.25 and 0.17), and ECG (6.76 and 0.26) in the cardiac and soleus muscles, respectively. The LDH activity (in 1 Mm/protein) in soleus muscle was smaller (P < 0.05) for ELG (0.33) compared to ECG (0.97), RLG (0.79), and RCG (1.07). For cardiac muscle, the LDH activity was smaller (P < 0.05) in ELG (1.38) compared to ECG (1.91) and RCG (2.55). The ECG and ELG showed increases in the maximum speed and a shift of the AT to higher effort levels after the training period, but no differences occurred between the exercised groups. In conclusion, the aerobic treadmill training combined with LLLT promotes an increase of oxidative capacity in this rat model, mainly in muscles with greater aerobic capacity.

Changes of DNA quality and meat physicochemical properties in bovine supraspinatus muscle during microwave heating.

BACKGROUND: The responses of foods to microwave exposure are usually evaluated only in terms of physicochemical properties, thus undervaluing the importance of DNA in an authentication process by methods based on polymerase chain reaction (PCR). In this study, the time effect of microwave heating on some meat physicochemical properties and DNA quality has been investigated. RESULTS: Cooking loss, instrumental colour, pH and other physicochemical parameters varied significantly during microwave cooking, reaching the lowest/highest values after 2.5 min of cooking. The exposure of meat to microwaves was found to affect characteristically the quality of extracted DNA (i.e. yield, purity and degradation). PCR products of both mitochondrial and nuclear regions were successfully observed in all samples. However, the band for large fragments became progressively fainter as treatment time increased. CONCLUSIONS: Microwave heating caused physicochemical changes in bovine supraspinatus muscle and influenced characteristically the yield and integrity of the extracted DNA, indicating that an accurate DNA quantification and a rational choice of the genes (i.e. mtDNA versus nDNA, fragment size, etc.) to be amplified are fundamental in an authentication process by PCR-based methods.

Ultraviolet irradiation alters the density of inner mitochondrial membrane and proportion of inter-mitochondrial junctions in copepod myocytes.

The efficient production of energy via oxidative phosphorylation is essential to the growth, survival, and reproduction of eukaryotes. The behavior (position of, and communication between, mitochondria) and morphology of mitochondria play key roles in efficient energy production and are influenced by oxidative stressors such as ultraviolet (UV) radiation. We tested the hypothesis that mitochondria change their behavior and morphology to meet energetic demands of responding to changes in oxidative stress. Specifically, we predicted that UV irradiation would increase the density of inner mitochondrial membrane and proportion of inter-mitochondrial junctions to influence whole-animal metabolic rate. Using transmission electron microscopy, we found that both three and six hours of UV-A/B irradiation (0.5 W/m2) increased the proportion of inter-mitochondrial junctions (with increasing mitochondrial aspect ratio) and the density of inner mitochondrial membrane in myocytes of Tigriopus californicus copepods. Mitochondrial density increased following both irradiation treatments, but mitochondrial size decreased under the six hour treatment. Metabolic rate was maintained under three hours of irradiation but decreased following six hours of exposure. These observations demonstrate that the density of inner mitochondrial membrane and proportion of inter-mitochondrial junctions can play formative roles in maintaining whole-animal metabolic rate, and ultimately organismal performance, under exposure to an oxidative stressor.

Low-intensity laser irradiation improves the mitochondrial dysfunction of C2C12 induced by electrical stimulation.

OBJECTIVE: We investigated the effects of electrical stimulation and low-intensity laser (LIL) energy on the mitochondrial function of cultured C2C12 myotubes in order to find a dosage that could be used to improve the function of mitochondria, and then rehabilitate exercise-induced damage and fatigue. BACKGROUND DATA: Many other studies in the past demonstrated that LIL had a cytoprotective effect, and a recent study also found that LIL could reduce muscular fatigue during tetanic contractions in rats. METHODS: Cultured C2C12 myotubes were subjected to electrical stimulation or/and LIL irradiation at various intensities. Reactive oxygen species (ROS) were detected with a fluorescent probe (DCFH-DA) and mitochondrial function was assessed with an MTT assay. RESULTS: The results showed that electrical stimulation at 20 ms, 5 Hz, and 45 V for 75 min can induce mitochondrial dysfunction in cultured C2C12 myotubes. Electrical stimulation-induced mitochondrial dysfunction was improved, but degeneration occurred with LIL at doses of 0.33-8.22 and 11.22-14.16 J/cm2, respectively, and these changes were markedly increased with LIL at 0.33 and 1.34 J/cm2, respectively. CONCLUSIONS: We conclude that treatment of myotubes with the proper dosage of LIL irradiation significantly diminished production of ROS and restored mitochondrial function, and this may provide a foundation for the use of photobiomodulation to treat exercise-induced mitochondrial dysfunction or skeletal muscular fatigue.

γ-Rays-generated ROS induce apoptosis via mitochondrial and cell cycle alteration in smooth muscle cells.

PURPOSE: γ-rays (IR) cause an increase in intracellular calcium [Ca(2+)], alters contractility and triggers apoptosis via the activation of protein kinase C in intestinal guinea pig smooth muscle cells. The present study investigated the role of the mitochondria in these processes and characterized proteins involved in IR-induced apoptosis. MATERIALS AND METHODS: Intestinal smooth muscle cells were exposed to 10-50 Gy from a (60)Co γ-source. Reactive oxygen species (ROS) levels were measured by colourimetry with a fluorescente probe. Protein expression was analyzed by immunoblotting and immunofluorescence. RESULTS: Apoptosis was inhibited by glutathione, possible by inhibiting the generation or scavenging ROS. Apoptosis was mediated by the mitochondria releasing cytochrome c leading to caspase 3 activation. IR increased the expression of the cyclins A, B2 and E and led to unbalanced cellular growth in an absorption dose-dependent manner. However, radiation did not induce alterations in the mitochondrial ultrastructure or in transmembrane electric potential. In contrast, IR increased the nuclear expression of cytoplasmic proteins and cyclins A and E. CONCLUSION: Smooth muscle cells subjected to IR undergo mitochondrial-mediated apoptosis that involves oncoproteins activation and preserves mitochondrial structure. IR also cause alterations in the expression and localization of both pro- and anti-apoptotic proteins.

In vitro photodynamic therapy with chlorin e6 leads to apoptosis of human vascular smooth muscle cells.

Percutaneous coronary intervention has become the most common and widely implemented method of heart revascularization. However, the development of restenosis remains the major limitation of this method. Photodynamic therapy (PDT) recently emerged as a new and promising method for the prevention of arterial restenosis. Here the efficacy of chlorin e6 in PDT was investigated in vitro using human vascular smooth muscle cells (TG/HA-VSMCs) as one of the cell types crucial in the development of restenosis. PDT-induced cell death was studied on many levels,including annexin V staining, measurement of the generation reactive oxygen species (ROS) and caspase-3 activity,and assessment of changes in mitochondrial membrane potential and fragmentation of DNA. Photosensitization of TG/HA-VSMCs with a 170 lM of chlorin e6 and subsequent illumination with the light of a 672-nm diode laser(2 J/cm2) resulted in the generation of ROS, a decrease in cell membrane polarization, caspase-3 activation, as well as DNA fragmentation. Interestingly, the latter two apoptotic events could not be observed in photosensitized and illuminated NIH3T3 fibroblasts, suggesting different outcomes of the model of PDT in various types of cells. The results obtained with human VSMCs show that chlorin e6 may be useful in the PDT of aerial restenosis, but its efficacy still needs to be established in an animal model.

Evaluation of mitochondrial respiratory chain activity in muscle healing by low-level laser therapy.

BACKGROUND: Recent studies demonstrate that low-level laser therapy (LLLT) modulates many biochemical processes, especially the decrease of muscle injures, the increase in mitochondrial respiration and ATP synthesis for accelerating the healing process. OBJECTIVE: In this work, we evaluated mitochondrial respiratory chain complexes I, II, III and IV and succinate dehydrogenase activities after traumatic muscular injury. METHODS: Male Wistar rats were randomly divided into three groups (n=6): sham (uninjured muscle), muscle injury without treatment, muscle injury with LLLT (AsGa) 5J/cm(2). Gastrocnemius injury was induced by a single blunt-impact trauma. LLLT was used 2, 12, 24, 48, 72, 96, and 120 hours after muscle-trauma. RESULTS: Our results showed that the activities of complex II and succinate dehydrogenase after 5days of muscular lesion were significantly increased when compared to the control group. Moreover, our results showed that LLLT significantly increased the activities of complexes I, II, III, IV and succinate dehydrogenase, when compared to the group of injured muscle without treatment. CONCLUSION: These results suggest that the treatment with low-level laser may induce an increase in ATP synthesis, and that this may accelerate the muscle healing process.

[Influence of N-acylethanolamines on embryonic cell culture survival under the effect of irradiation].

The effects of the N-stearoylethanolamine (NSE), N-oleoylethanolamine (OEA) and N-acylethanolamine (NAE) mixture on the cell survival, apoptosis and activity of mitochondrial succinate dehydrogenase (SDG) and glycerophosphate dehydrogenase (GFDG) in embryonic cell culture under normal conditions and irradiation were compared in the work. It was shown, that all investigated NAE were able to modulate the proliferative activity of intact cells as well as irradiation-exposed cells in concentration-depended manner. The most pronounced effect was observed under the NAE mixture action. It was established, that NAE prevented the damage effects of the irradiation and diminished the activation of apoptotic cell death. It was found that NSE and OEA decreased the activity of the SDG (42.3 and 44.14%, accordingly) and the GFDG activity (14.67 and 17.33%, accordingly) in embryonic cell culture, while addition of the NAE mixture decreased SDG activity by 20%, at the same time GFDG activity increased by 20%. Our findings suggested that antiproliferative effects of NAE depended on their influence on mitochondrial functioning.

Argon laser microirradiation of mitochondria in rat myocardial cells in tissue culture. VII. fibrillation in ventricle and auricle cells.

Rat myocardial cells in vitro were irradiated in individual mitochondria with an argon ion laser microbeam. The contractile respone termed fibrillation in single and multicellular groups of both ventricle and auricle cells were compared. Specific correlations were made between fibrillation duration, the number of cells in the group, and the number of times the cells had fibrillated. Correlations were also made for the number of laser shots needed to induce fibrillation and the number of cells in the group. Another set of correlations were made between the pre-irradiation beat frequency and the beat frequency following recovery. Several differences and similarities of the above parameters were detected between auricle and ventricle cells. A comparison of the morphology and ultrastructure of auricle and ventricle cells also revealed significant differences.

[Ultrastructure of skeletal muscle tissue of microwave damaged chick embryos]. / Ul'trastruktura skeletnoi myshechnoi tkani kurinykh émbrionov pri mikrovolnovom porazhenii.

The effect of ultrahigh-frequency energy of electromagnetic field of low intensity on certain morphometric indices of intracellular organoids and on cellular ultrastructure were studied in the developing skeletal muscular tissue of chick embryos. In the skeletal muscles of irradiated embryos a limited cellular destruction, structural disorders in myonic organoids were revealed. Reactive-recovery processes manifested themselves in hyperplasy and hypertrophy of organoids, in activation of protein synthesis, in increasing amount of myosatellites. At early stages after irradiation peripheral mitochondria are subjected to greater changes. After hatching, central mitochondria in myons suffer more. Quantitative changes in myonic organoids and degree of their destruction seem to depend on.a peculiar differentiation of the muscular tissue, blood supply and innervation of the muscle as an organ.