[Effects of Light Near-Infrared Radiation on Rats Assessed by Succinate Dehydrogenase Activity in Lymphocytes on Blood Smears].

Biological effects of light near infrared radiation (850 nm), with modulation acoustic frequency of 101 Hz, was studied. The study was conducted on rats, the effect was recorded by succinate dehydrogenase activity in lymphocytes on the blood smear after administration of the activating dose of adrenaline, which simulates the state of the organism in the early stages of the pathogenic effects (stress). A pronounced regulating effect of infrared radiation on the activity of succinate dehydrogenase in animals activated by adrenaline was shown. Infrared radiation has a normalizing effect reducing the degree of inhibition or activation of the enzyme induced by adrenaline and had no effect on the control animals. Thus, by modulating the activity of succinate dehydrogenase infrared radiation regulates energy production in the mitochondria supported by the most powerful oxidation substrate--succinic acid, which is especially pronounced under stress.

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.

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

Laser therapy is used in many biomedical sciences to promote tissue regeneration. Many studies involving low-level laser therapy have shown that the healing process is enhanced by such therapy. In this work, we evaluated mitochondrial respiratory chain complexes II and IV and succinate dehydrogenase activities in wounds after irradiation with low-level laser. The animals were divided into two groups: group 1, the animals had no local nor systemic treatment and were considered as control wounds; group 2, the wounds were treated immediately after they were made and every day after with a low-level laser (AsGa, wavelength of 904 nm) for 10 days. The results showed that low-level laser therapy improved wound healing. Besides, our results showed that low-level laser therapy significantly increased the activities of complexes II and IV but did not affect succinate dehydrogenase activity. These findings are in accordance to other works, where cytochrome c oxidase (complex IV) seems to be activated by low-level laser therapy. Besides, we showed, for the first time, that complex II activity was also activated. More studies are being carried out in order to evaluate other mitochondrial enzymes activities after different doses and irradiation time of low-level laser.

Extracellular environment as one mediator of blue light-induced mitochondrial suppression.

OBJECTIVES: The current study tested the hypothesis that the extracellular environment mediates mitochondrial suppression of oral epithelial cells and fibroblasts by blue light. METHODS: We exposed Balb fibroblasts (Balb), normal human epidermal keratinocytes (NHEK), and oral squamous carcinoma cells (OSC2) to blue light (30-120J/cm2) in different cell-culture media and in phosphate buffered saline (PBS). Mitochondrial activity (MTT method) was used to assess cellular response 72 h post-light exposure. Cell-culture media were replaced or supplemented before or after light exposure to assess the variables of exposure time and medium degradation as mediators of blue light-induced effects. RESULTS: Mitochondrial activity of NHEK was not suppressed by exposure to blue light regardless of extracellular conditions. The mitochondrial activity of OSC2 and Balb cells was suppressed most when cells were exposed to light in cell-culture medium (versus PBS). Blue light suppressed mitochondrial activity more when irradiated medium remained in contact with the cells at least 1h, indicating a time-dependence of the medium effects. Neither a replacement nor a supplementation of medium components reduced blue light-induced mitochondrial suppression. SIGNIFICANCE: Our results suggest that tissue environments influence cellular responses to blue light and that these environments should be considered when assessing any biological effects of blue light during the photopolymerization of restorative resins.

Blue light differentially alters cellular redox properties.

Blue light (lambda = 380-500 nm) historically has been used to initiate polymerization of biomaterials and recently has been proposed as a therapeutic agent. New evidence suggests that cell-type-specific responses result from redox changes induced by exposure to blue light. Cultured cells were exposed to defined doses of blue light, equivalent to exposure times of 10 s and 2 min, to achieve energies of 5 J/cm2 and 60 J/cm2, respectively, after which (a) viable cell number, (b) cellular protein profiles, (c) mitochondrial succinate dehydrogenase (SDH) activity, (d) total reactive oxygen species (ROS), and (e) induction of apoptosis were compared to that of nonexposed control cultures. Results showed that blue-light exposure arrested monocyte cell growth and increased levels of peroxiredoxins. SDH activity of normal epidermal keratinocytes (NHEK) was slightly enhanced by blue light, whereas identical treatment of OSC2 oral tumor cells resulted in significant suppression of SDH activity. Blue-light exposure generally induced higher levels of total ROS in OSC2 cells than in NHEK. Finally, only OSC2 cells exhibited signs of apoptosis via Annexin V staining following exposure to blue light. These data support the central hypothesis that blue light induces an oxidative stress response in cultured cells resulting in cell-type-specific survival outcomes. The identification of oxidative stress as a mediator of the effects of blue light is a critical first step in defining its biological risks and therapeutic opportunities.

Blue light differentially modulates cell survival and growth.

Previous studies have reported that blue light (400-500 nm) inhibits cell mitochondrial activity. We investigated the hypothesis that cells with high energy consumption are most susceptible to blue-light-induced mitochondrial inhibition. We estimated cell energy consumption by population doubling time, and cell survival and growth by succinate dehydrogenase (SDH) activity. Six cell types were exposed to 5 or 60 J/cm(2) of blue light from quartz-tungsten-halogen (QTH), plasma-arc (PAC), or argon laser sources in monolayer culture. Post-light SDH activity correlated positively with population doubling time (R(2) = 0.91 for PAC, 0.76 for QTH, 0.68 for laser); SDH activity increased for cell types with the longest doubling times and was suppressed for cell types with shorter doubling times. Thus, light-induced exposure differentially affects SDH activity, cell survival, and growth, depending on cell energy consumption. Blue light may be useful as a therapeutic modulator of cell growth and survival.

Mitochondrial respiratory chain features after gamma-irradiation.

Radiation provokes damage to DNA but also to membrane and protein structure. Radiolysis is a tool used very often in the study of free radical biological effects and of scavenger molecules effectiveness. Nitroimidazoles have been demonstrated to enhance the radiation effects on biological structures. The studies we have performed on isolated mitochondria irradiated, with and without nitroimidazoles, at a radiation dose equal to LD90, indicate that this treatment is not able to affect the structural and functional features investigated (ubiquinone-10, fatty acids, respiratory cytochrome levels or membrane fluidity and respiratory enzymatic activities), suggesting that an involvement of such externally produced radicals on membrane damage is unlikely. Moreover it was ascertained that the mitochondrial redox activities do not take part into the intracellular nitroimidazole reduction.

[The significance of the hypoxic factor in adaptation to the action of ionizing radiation as a result of the accident at the Chernobyl Atomic Electric Power Station]. / Pro znachennia hipoksychnoho faktora v adaptatsiï do diï ionizuiuchoho vyprominiuvannia vnaslidok avariï na ChAES.

Data are presented from cytochemical study of the neutrophil myeloperoxidase oxygen dependent enzymes and lymphocyte succinate dehydrogenase in the peripheral blood of 2169 persons subjected to follow-ups of many kinds, at year 7 and 8 after the Chernobyl accident. By comparison with early time periods, the process of normalization of the measured values has been found to be the case, with SDG returning to normal earlier than MPO. There was an increase in the number of individuals with reduced MPO activity and stable percentage of those demonstrating increased SDG activity (30-42% versus 12%). Significance of the adaptation process in the time course of changes of the values measured is discussed along with role of hypoxic factor in adaptation to the ionizing irradiation exposure after the Chernobyl accident.

[Functional interrelationships between dehydrogenases and transaminases in various regions of the rat brain as affected by small doses of gamma-radiation]. / Funktsional'nye vzaimootnosheniia mezhdu degidrogenazami i transaminazami v razlichnykh otdelakh golovnogo mozga krys pri deistvii malykh doz gamma-izlucheniia.

Wistar rats were exposed to 10, 20 and 40 sGy of gamma-irradiation. Maximal activity of 2-oxoglutarate- and succinate dehydrogenases, as well as of aspartate-2-oxoglutarate and pyruvate-2-oxoglutarate in the brain tissue was measured within 30 days after the exposure. Dehydrogenase activity was measured in the brain cortex, limbic system and in the cerebellum; transaminase activity was determined in the brain cortex, diencephalic zone and in the hypophysis. It was found that small doses of irradiation resulted in an oscillating increase of dehydrogenase activity with a decrease of the amplitude by the 30th day of the experiment. As the dose of irradiation increases, the periods of normal activity change into phases of inhibition of dehydrogenase action, the period of oscillations diminishes. Transaminase activity, as a rule, changes in a phase in a reversible manner as compared with dehydrogenase activity. These observations are discussed from the point of view of structural relations between these two systems in the intramitochondrial supramolecular structures, the so-called metabolons.

[The effect of a low-intensity helium-neon laser on succinate dehydrogenase activity in unmineralized periodontal and dental tissues]. / Vliianie nizkointensivnogo gelii-neonovogo lazera na aktivnost' suktsinatdegidrogenazy v nemineralizovannykh tkaniakh parodonta i zubov.

Laser effects with energy density of 28.8, 86.4, 144 J/cm2 and 86.4 J/cm2 in combination with fluorine agents on succinate dehydrogenase (SDH) activity were studied in 264 noninbred white rats (5 groups). The animals were decapitated in 1, 6, 12, 24, 48, 72 h and 12 and 30 days. The findings evidence elevated SDH activity virtually in all periods tested. Laser exposure enhanced enzymic activity in all the animals.

[Metabolic reactions in sensory and motor neurons after exposure to various physical factors]. / Metabolicheskie reaktsii v chuvstvitel'nykh i dvigatel'nykh neironakh pri deistvii na organizm nekotorykh fizicheskikh faktorov.

A course of radiation of biologically active points of the guinea pig lumbosacral area has been performed by means of focused frequency-modulated and a continuous beam of the helium-neon laser, having various power density, as well as by means of an impulsive magnetic field. A comparative histoenzymatic analysis of sensitive neurons of the lumbar spinal nodes and of the caudal-mesenteric motor neurons has been carried out. The response of the metabolic processes in the sensitive neurons depends not so much on the type of action, as on adequacy of the parameters applied. The power density, frequency of impulse generation and time of application are important for characterization of the adequacy of the laser radiation. Peculiarities of the metabolic reactions in the sensitive and sympathetic neurons in response to the actions studied are discussed.

[Various indicators of brain metabolism during hypoxia and hyperthermia]. / Nekotorye pokazateli metabolizma v golovnom mozge pri gipoksii i perenagrevanii.

Hypoxic, thermal and radiation tolerance of rats exposed to hypoxia and overheating for 4 to 22 days was measured. Activities of succinic dehydrogenase, lactate dehydrogenase and alkaline phosphomonoesterase as well as water content in the brain were examined biochemically and histochemically. Brain tolerance and metabolism varied in a phasic manner. Both specific and nonspecific adaptive reactions were identified. A direct correlation was established between hypoxic tolerance and aerobic oxidation decrease as well as enzyme-dependent transport in cerebral vascular walls. A relationship between CNS radiation tolerance and permeability of the blood-brain barrier was detected.

In vitro photosensitization of tumour cell enzymes by photofrin II administered in vivo.

The ability of injected Photofrin II, a preparation enriched in hydrophobic dihaematoporphyrin ethers and esters, to photosensitize selected mitochondrial and cytosolic enzymes during illumination in vitro was examined. Preparations of R3230AC mammary tumours, obtained at designated times after a single dose of Photofrin II, displayed a time-dependent photosensitivity. Maximum inhibition of mitochondrial enzymes occurred at 24 hours post-treatment, whereas no inhibition of the cytosolic enzyme, pyruvate kinase, was observed over the 168 hour time course. At the selected 24 hour time point, mitochondrial enzyme photosensitisation was found to be drug dose (5.25 mg kg-1 Photofrin II) and light dose dependent, the rank order of inhibition being cytochrome c oxidase greater than F0F1 ATPase greater than succinate dehydrogenase greater than NADH dehydrogenase. We conclude that porphyrin species contained in Photofrin II accumulate in mitochondria of tumour cells in vivo and produce maximum photosensitisation at 24-72 hours after administration to tumour-bearing animals. The time course observed here with Photofrin II is similar to that seen previously with the more heterogenous haematoporphyrin derivative preparation in this in vivo-in vitro model.

[Changes in the activity of the oxidative and hydrolytic enzymes of the animal cerebral cortex in the early periods after gamma irradiation and the use of meksamine and the synthetic analog of prostaglandin F2 alpha-estrofan]. / Izmenenie aktivnosti okislitel'nykh i gidroliticheskikh fermentov korygolovnogo mozga zhivotnykh v rannie sroki posle gamma-oblucheniia v usloviiakh primeneniia meksamina i sinteticheskogo analoga prostaglandina F2 al'fa-éstrofana.

In experiments with (CBA x C57B1/6)F1 mice it was shown that LDH activity moderately increased 5 min after exposure of the head to 200 Gy gamma radiation. After 60 min, there was a 24.4 per cent decrease in alkaline phosphatase activity and a 24.3 per cent increase in SDG activity. Injected prior to irradiation meksamine precluded the postirradiation increase in SDH and alleviated the postirradiation decrease in alkaline phosphatase.

Modification of bovine heart succinate dehydrogenase with ethoxyformic anhydride and rose bengal: evidence for essential histidyl residues protectable by substrates.

Purified and membrane-bound succinate dehydrogenase (SDH) from bovine heart mitochondria was inhibited by the histidine-modifying reagents ethoxyformic anhydride (EFA) and Rose Bengal in the presence of light. Succinate and competitive inhibitors protected against inhibition, and decreased the number of histidyl residues modified by EFA. The essential residue modified by EFA was not the essential thiol of SDH, but modification of the essential thiol abolished the protective effect of malonate against inhibition of SDH by EFA. The EFA inhibition was reversed by hydroxylamine nearly completely when the inhibition was less than or equal to 35%, and only partially when the inhibition was more extensive. The uv spectrum of EFA-modified SDH before and after hydroxylamine treatment suggested that extensive inhibition of SDH with EFA may result in ethoxyformylation at both imidazole nitrogens of histidyl residues. Such a modification is not reversed by hydroxylamine. Succinate dehydrogenases and fumarate reductases from several different sources have similar compositions, and the two enzymes from Escherichia coli have considerable homology in the amino acid composition of their respective flavoprotein and iron-sulfur protein subunits. In the former, there is a short stretch containing conserved histidine, cysteine, and arginine residues. These residues, if also conserved in the bovine enzyme, may be the essential active site residues suggested by this work (histidine) and previously (cysteine, arginine).

Hematoporphyrin derivative-induced photosensitivity of mitochondrial succinate dehydrogenase and selected cytosolic enzymes of R3230AC mammary adenocarcinomas of rats.

The photosensitizing activity of hematoporphyrin derivative (HPD) was investigated by studying selected enzymes localized to mitochondria and cytosol of R3230AC mammary adenocarcinomas. Experiments in vitro demonstrated that mitochondrial succinate dehydrogenase was inhibited in a drug dose- and light exposure time-related manner; at 7.0 micrograms of HPD per ml or higher, enzyme activity was inhibited greater than 50% after 15 min of photoradiation. The three cytosol enzymes studied under the same conditions in vitro demonstrated different photosensitivities. Pyruvate kinase activity was significantly inhibited in a dose- and time-related fashion, whereas lactate dehydrogenase was inhibited to a lesser extent, and glucose phosphate isomerase activity was inhibited only at the highest dose (70 micrograms of HPD per ml) used. The time-course of these responses was examined with an in vivo-in vitro protocol, consisting of photoradiation of mitochondria and cytosol prepared from tumors obtained at various times (up to 1 week) after a single injection of HPD (80 mg/kg). Pyruvate kinase activity was markedly inhibited at early times returning to initial levels by 48 hr; neither lactate dehydrogenase nor glucose phosphate isomerase was inhibited by this treatment. Mitochondrial succinate dehydrogenase and cytochrome c oxidase activities displayed significant photoradiation-induced inhibitions, with greatest inhibition occurring between 24 and 96 hr after injection of HPD; at 1 week, succinate dehydrogenase activity had returned to its initial level, but cytochrome c oxidase activity remained significantly inhibited. These data suggest that HPD-induced photosensitization of mitochondria may be an important site of action contributing to tumor cell cytotoxicity and regression as a result of photoradiation therapy.