[Loosening of condensed chromatin in human blood lymphocytes exposed to irradiation with a low-energy He-Ne laser].

It was shown that, 1 h after irradiation of human blood lymphocytes with a He-Ne laser at 56 J/m2 (5.6 W/m2, 10 s), the relative optical density of condensed chromatin masses observed in ultrathin sections was decreased (p < 0.01); i.e., the condensed chromatin became less compact. Such transition of condensed chromatin to a more "open" state may improve its availability for regulatory proteins and transcriptional factors. The irradiation also results in dispersion of condensed chromatin clumps in the nucleoplasm and enhancement of their angularity, i.e., in extension of the clump surface. These shifts, correlating with the activation of transcription, may be due to decompaction of the chromatin fibers not only on the periphery of chromatin clusters in the center of the nucleus, but also within the masses of condensed chromatin.

Exact action spectra for cellular responses relevant to phototherapy.

OBJECTIVE: The aim of the present work is to analyze available action spectra for various biological responses of HeLa cells irradiated with monochromatic light of 580-860 nm. BACKGROUND DATA: Phototherapy (low-level laser therapy or photobiomodulation) is characterized by its ability to induce photobiological processes in cells. Exact action spectra are needed for determination of photoacceptors as well as for further investigations into cellular mechanisms of phototherapy. METHODS: Seven action spectra for the stimulation of DNA and RNA synthesis rate and cell adhesion to glass matrix are analyzed by curve fitting, followed by deconvolusion with Lorentzian fitting. Exact parameters of peak positions and bandwidths are presented. RESULTS: The peak positions are between 613.5 and 623.5 nm (in one spectrum, at 606 nm), in the red maximum. The far-red maximum has exact peak positions between 667.5 and 683.7 nm in different spectra. Two near infrared maxima have peak positions in the range 750.7-772.3 nm and 812.5-846.0 nm, respectively. CONCLUSIONS: In the wavelength range important for phototherapy (600-860 nm), there are four "active" regions, but peak positions are not exactly the same for all spectra.

[Structure of mitochondria and activity of their respiratory chain in subsequent generations of yeast cells exposed to He-Ne laser light].

The data on the effect of He-Ne laser light (lambda = 632.8 nm) on mitochondria of yeasts in late log phase were reviewed. The qualitative analysis of the ultrathin sections of cells demonstrated a nonuniform thickness of the giant branched mitochondria typical for budding yeasts. Exposure to a dose of 460 J/m@2 accelerated cell proliferation, activated respiratory chain enzymes (cytochrome c oxidase and NADH dehydrogenase), and also changed the microstructure of the giant mitochondria--much of the narrow regions of the mitochondrial tube with sections < or = 0.06 microm2 were dilated (while no signs of organelle damage were observed). Relative surface area of the cristae increased in such mitochondria, which can be due to the activation of their respiration and ATP synthesis. The number of associations between mitochondria and endoplasmic reticulum increased in cells in early log phase, which reflects high capacity of mitochondria to absorb Ca2+. Altered giant mitochondria configuration can increase the efficiency of both energy transfer and Ca2+ distribution in the cytoplasm.

[Increase in the number of contacts of endoplasmic reticulum with mitochondria and plasma membrane in yeast cells stimulated to division with He-Ne laser light]. / Uvelichenie kolichestva assotsiatsii éndoplazmaticheskogo retikuluma s mitokhondriiami i plazmaticheskoi membranoi v drozhzhevykh kletkakh, stimulirovannykh k deleniiu svetom He-Ne-lazera.

We studied the ultrastructure of Torulopsis sphaerica yeast cells irradiated with He-Ne laser (lambda = 632.8 nm, dose--460 J/m2) and then cultured for 6 h in the nutrient with 1% glucose by aeration. The length of membranes of endoplasmic reticulum (ER) and the number of its associations with mitochondria (M) and plasma membrane (PM) were measured on ultrathin sections. A distance of less than 50 nm between heterogeneous membranes was considered as an "association". The cells from irradiated cultures are characterized by the following features: 1) the length of cortical ER membranes in relation to cellular perimeter, and the length of perinuclear ER membranes in relation to nuclear perimeter increase, resp., by 21 and 79%; 2) the number of ER-PM associations per cellular section, and that per unit of PM length increase, resp., by 26 and 41%; 3) the number of ER-M association in relation to the total mitochondrial perimeter, and to perimitochondrial ER increase by 80 and 87%, resp. The latter may be associated with Ca2+ uptake by mitochondria associated with ER, which results in activation of respiration and ATP production.

[Morphometric study of yeast cells of Torulopsis sphaerica after He-Ne-laser irradiation]. / Mopfometricheskoe issledovanie drozhzhevykh kletok Torulopsis sphaerica posle oblucheniia svetom He-Ne-lazera.

A study was made of structural organization of Torulopsis sphaerica cells irradiated with He-Ne (lambda = 632.8 nm; dose--460 J/m2) and then cultured in the nutrient with 1% glucose and O2 for 6 h. The computer analysis of electron images of cell sections was carried out. Evidences of stimulation of cell proliferation were found, including decrease in the areas of cell and chondriome profiles, decrease in the number of mitochondria on sections, elongation of cells and mitochondria, and increased variability of cell parameters. In addition, cells of irradiated cultured were characterized by the increase in the number of mitochondria contacting the endoplasmic reticulum (in this case the outer mitochondrial membrane presumably associates with the ER membrane), which may suggest the activation of ATP synthesis. Thus, He-Ne laser irradiation activates cell metabolism even at the early stage of culture growth.

Cell attachment to extracellular matrices is modulated by pulsed radiation at 820 nm and chemicals that modify the activity of enzymes in the plasma membrane.

BACKGROUND AND OBJECTIVES: Adhesive interactions between cells and extracellular matrices play a regulative role in wound repair processes. The objective of this investigation is to study action mechanisms of pulsed radiation at 820 nm on cellular adhesion in vitro. Light emitting diodes (LED) at 820 nm are widely used for treatment of wounds of various etiology. STUDY DESIGN/MATERIALS AND METHODS: The LED (820 +/- 10 nm, 10 Hz, 16-120 J/m(2)) is used for the irradiation of HeLa cell suspension. In parallel experiments, amiloride (5 x 10(-4) M), ouabain (7 x 10(-5) M, 7 x 10(-4) M), quinacrine (6 x 10(-4) M), arachidonic acid (1 x 10(-5) M), glucose (2 x 10(-4) M), and ATP (5 x 10(-5) M) are added to the cell suspension before or after the irradiation procedure. The cell-glass adhesion is studied using the adhesion assay technique described in Lasers Surg Med 1996; 18:171. RESULTS: Cell-glass adhesion increases in a dose-dependent manner following the irradiation. Preirradiation eliminates the inhibition of cell attachment caused by ouabain, arachidonic acid, and ATP. The inhibitive effect of quinacrine on the cell attachment is eliminated by the irradiation performed after the treatment with the chemical. Irradiation and amiloride have a synergetic stimulative effect on the cell attachment. The threshold dose for the cell attachment stimulation by the irradiation is decreased by the treatment of the cell suspension with amiloride or ouabain. CONCLUSIONS: The results obtained indicate that pulsed IR radiation at 820 nm increases the cell-matrix attachment. It is the modulation of the monovalent ion fluxes through the plasma membrane and not the release of arachidonic acid that is involved in the cellular signaling pathways activated by irradiation at 820 nm. Preirradiation has a protective effect against the inhibitive action of ouabain, arachidonic acid, ATP, and quinacrine on cell attachment process. It is supposed that irradiation activates those signaling pathways in cells which attenuate the inhibitive action of these chemicals.

Donors of NO and pulsed radiation at lambda = 820 nm exert effects on cell attachment to extracellular matrices.

The adhesion of HeLa cells to a glass matrix was evaluated after the irradiation of the cell suspension with a pulsed near-infrared light-emitting diode (lambda = 820 nm, frequency 10 Hz, dose 8-120 J/m(2)) and treatment with two donors of nitric oxide, sodium nitroprusside (SNP, 5 x 10(-4) M) and NaNO(2) (4 x 10(-4) M). It was found that in an irradiated cell suspension, the cell-glass adhesion increases in a dose-dependent manner (a bell-shaped curve with a maximum at 60 J/m(2)). The treatment of cells with SNP or NaNO(2) before the irradiation eliminates the radiation-induced attachment stimulation. Pretreatment of cells with SNP not only eliminates the radiation-induced attachment stimulation but inhibits the attachment of irradiated (but not non-irradiated) cells. It is suggested that a modulation of the activity of respiratory chain (probably the alteration of the activity of cytochrome c oxidase) is involved in radiation-induced increase of cell attachment.

Cell attachment modulation by radiation from a pulsed light diode (lambda = 820 nm) and various chemicals.

BACKGROUND AND OBJECTIVE: Adhesive interactions between cells and extracellular matrices play a regulative role in wound repair processes. The objective of this investigation is to study the mechanisms of light action on cellular adhesion in vitro. The adhesion of HeLa cells to a glass matrix is evaluated after irradiation with a pulsed near-infrared (IR) diode and treatment with various chemicals. STUDY DESIGN/MATERIALS AND METHODS: A semiconductor diode (820 +/- 10 nm, 10Hz, 16--120 J/m(2)) is used for irradiation of the cell suspension. In parallel experiments, various chemicals (mannitol, melatonin, ethanol, ascorbic acid, superoxide dismutase, catalase, rotenone, azide, dinitrophenol (DNP), methylene blue, and hydrogen peroxide) are added to the cell suspension before or after the irradiation procedure. The cell-glass adhesion is studied by using the adhesion assay technique (Lasers Surg. Med. 1996;18:171). RESULTS: It has been found that cell-glass adhesion increases in a dose-dependent manner after irradiation. The treatment of the cells with antioxidants (free radical scavengers), e.g., mannitol, melatonin, ethanol, and ascorbic acid, as well as with the ionophore DNP, eliminated the light effect. The respiratory chain inhibitors rotenone and azide strongly modified the light effect, depending on the dose. The oxidative agents hydrogen peroxide (in a low concentration) and methylene blue increased the cell adhesion. Superoxide dismutase did not modify the light effect. The effect of the catalase (stimulative or suppressive) was dependent on its concentration and treatment sequence. Preirradiation was found to decrease (or normalize to the control level) the suppressive effects of some chemicals. CONCLUSION: The results obtained are evidence that first, pulsed IR radiation with certain parameters modulates the cell-matrix attachment. second, free radical and redox processes are involved in the cell-matrix interaction, probably at some stage(s) of the photosignal transduction. Third, both types of the primary reactions in the respiratory chain, namely, the increase of the electron flow and production of the reactive oxygen species, cause a transient oxidative stress in the cytoplasm.

Nonmonotonic behavior of the dose dependence of the radiation effect on cells in vitro exposed to pulsed laser radiation at lambda = 820 nm.

BACKGROUND AND OBJECTIVE: In recent years, clinical low-intensity laser therapy practice has used pulsed radiation, mainly from semiconductor lasers. Experimental works devoted to the study of relationships between biological and clinical effects and parameters of pulsed radiation are practically absent. STUDY DESIGN/MATERIALS AND METHODS: The radiation source was a laser diode emitting at 820 nm (292 and 700 Hz, duty factor 80%; doses from 7 J/m2 to 5 x 10(5) J/m2; intensities 4, 12, 51, 152, 633, and 1,900 W/m2; irradiation time from 1 to 30 s). Four biological models were used: nucleated cells of murine spleen (splenocytes) and bone marrow (karyocytes), murine blood, and HeLa cells cultivated in vitro. The intensity of luminol-amplified chemiluminescence (in case of murine models) and the adhesion of HeLa cell membranes were measured as a function of the irradiation dose. RESULTS: Within the wide exposure dose range used we obtained seven maxima in the dose vs. biological effect curves: at fluences near 20, 1 x 10(2), 3 x 10(2), 8 x 10(2), 3 x 10(3), 1 x 10(4), and 3 x 10(4) J/m2. The peaks coincided for all four models. CONCLUSION: The dose curves obtained with different cellular systems are of the same type and are characterized by seven peaks in the dose interval studied (7 J/m2 to 5 x 10(5) J/m2).