The concept of blood irradiation therapy originates from Germany (K. Naswitis) and United States (E. Knott)' in the 1920s. In their works devices for extracorporeal inadiation of small amounts of blood with UV radiation were developed. The method was used to treat bacterial, viral and autoimmune diseases but a new wave of antibiotics and vaccines in 1 950s caused the method to fall into oblivion. Its second birth occurred in former USSR in the late 1 970s - early 1 980s and got a widespread use in hundreds of clinics as well as in vetermary medicine. Beside extracorporeal UV inadiation of blood Russian doctors started to use low-intensity He-Ne laser irradiation through a fiber and a needle (intravenous inadiation). Low-intensity He-Ne and later semiconductor lasers emitting in red - to - near IR region were also used for extracorporeal and over-venous inadiation. The rise of multidrug resistance of bacterial strains, strong side effects of some effective drugs, AIDS and hepatitis have originate the interest to methods of blood inadiation also in the West. Note that a blood inadiation device (extracorporeal inadiation with UV) has received a FDA clearance (51 0 (k)status), and there appeared an ultraviolet blood inadiation website (Dr. K. Dillon).

The aim of the present work was to investigate effect of low power laser irradiation (LPLI) on platelet function in vitro. He-Ne laser (Optronix, USA; (lambda) - 632.8 nm, output power - 7 mW) was employed. Platelet adhesion and aggregation in whole blood (WB) under defined shear conditions were assayed by a Cone and Plate(let) Analyzer. Platelet activation was evaluated by flow cytometry. Level of platelet cGMP was estimated by immunoenzyme assay. Experiments performed showed that LPLI of WB resulted in decrease of platelet deposition on extracellular matrix at high shear rate (1300 s^-1). Similar results were obtained using surfaces precoated with either collagen type I or von Willebrand factor. LPLI inhibited fibrinogen binding as well as P-selectin expression on the platelet membrane, induced by thrombin analogue. It was found out that primary acceptor of laser energy responsible for the effect on platelets was located in platelets themselves and not in blood plasma or in other blood cells. LPLI of gel- filtered platelets resulted in increase of intracellular level of cGMP both in the absence and in presence of izobutylmethylxantine (phosphodiesterase inhibitor) suggesting stimulation of synthesis rather than destruction of cGMP under the influence of LPLI. It is suggested that guanylate cyclase and/or NO-synthase might serve as primary acceptors of He-Ne laser light in platelets.

This paper deals with the cytofluorimetric investigation into an early reaction of T- and B-lymphocytes from human peripheral blood (accessibility of the overall nuclear chromatin to acridine orange) to irradiation with a He-Ne laser (632.8 nm, 67 J/m², 10 s). Our finding is that it is only the T-lymphocytes that react to the laser treatment, whereas a treatment with the mitogen phytohemagglutinin during one hour has an effect on both types of lymphocytes.

We have demonstrated recently that luminol- or lucigenin- dependent chemiluminescence (LM-CL and LC-CL, respectively) registered from whole non-diluted blood may reach rather high intensities. Here we demonstrate, that parameters of both LM-CL and LC-CL in non-diluted human blood during RB development is influenced by a reflective screen of aluminum foil surrounding a test tube with a blood sample. Photons reflected back to a sample accelerate slowly developing and retard rapidly development RB. As demonstrated by histochemical NBT test, wrapping samples with blood in aluminum foil effected also the reductive neutrophil activity in. These results indicate that weak light fluxes generated in the course of RB may influence physiological processes in blood. They also provide the basis for an explanation for the phenomenon of the interaction two blood samples in one of which RB in the presence of luminol was stimulated through the optical channel previously reported by us.

The influence of low-intensity linearly polarized visible ((lambda) equals 632.8 nm) and near infrared ((lambda) equals 797 nm) laser radiation on healing of skin wounds was compared histologically. Three round lesions measuring about 6 mm in diameter were created at the end of the spinal column of 20 rats divided in two groups of 10 animals by burning the rat back skin with liquid nitrogen. Lesions #1 and #2 were illuminated using He-Ne or GaAlAs laser radiation. The laser polarization was aligned with the rat spinal direction in lesion #1, lesion #2 with the perpendicular relative orientation and lesion #3 was not irradiated (control). The animals were irradiated with a total dose of 1 J/cm² on day 3, 8, 11 and 14. After each irradiation, two rats from each group were killed to obtain morphological information. On day 17 the last rats were killed. The irradiated lesions presented a significant acceleration on the wound repair compared to the control. The results showed that the relative orientation of the electric field has an essential role on the healing process if exposure is visible light. On the other hand, no morphological difference was observed between illuminated lesions by near infrared radiation with respect to polarization orientation.

Intracellular Ca concentration [Ca2 is ofparamount importance in the regulation of many cellular processes. The cytosolic-free calcium is buffered to approximately 100-150 nM Calcium oscillations arise either spontaneously'2 response to stimulation by extracellular signals such as hormones or neurotransmitters . Irradiation of endogenous or exogenous photosensitizers with visible or ulimviolet light is a way to cause an elevation in [Ca21, .This elevation was reported to be dependent on the concentration of photosensitizers, light dose and exiracellular cations . Prolonged mcubalion of the photosensitizer with the cell causes a persistent elevalion in [Ca211 following light irradiation, which is believed to be the early event in cell death in photodynamic therapy (PDT) .A short-time incubation with the photosensitizer followed by "°low-level light irradiation (LLL) causes only a iransient increase in the [Ca21,"'2 In previous studies we noticed that light at the same fluences which cause small [Ca?1 oscillation 13 induces acceleration in cell mitosis, while with higher doses of light cell proliferation was inhibited'4 j general, it seems that whereas high light intensity (over 20 JIcm2)causes irreversible damage to the cells, low level light (LLL) irradiation causes biostiinulation. Thus, there are numerous examples where LLL irradiation has been beneficial in dermatology, gynecology, otorhinolaryngology and other medical areas1549 In addition to the intensity dependent elevation of calcium in response to photostimulation, there is accumulating evidence concerning the generation of reactive oxygen species (ROS) in cell association with light absorbency2026 j the photohemolysis process of human eiythrocytes caused by PDT using bacteriocklorin a as sensitizer, generation of hydroxyl radical (.OH) and singlet oxygen ('02) was observed26 Singlet oxygen '02 was also found in Friend Erythroleukaemic cells after irradiation with HeNe laser without using photosensitizers 24 ROS are toxic to the cell 21 concenirations, it is believed that low concenirations have a positive biostimulalive effect ROS were repoited to stimulate cultured fibroblasts to proliferate 25 and were found to play an important role in the acquisition of fertilizing ability 24,27 Consequently, considerable attention has been paid to the generation and the role of ROS in cell development Consideiable attention was also given to the changes in [Ca21, in response to chemicals and irradiation. However there are not enough reports dealing with the relation between ROS production and [Ca211 changes, in particular the connection between ROS, [Q2J biostimnlation. We have compared free iniracellular 2+ concentration changes in cardiac myocytes after visible light irradiation and upon exposure to low concentration 11202. Hydrogen peroxide is relatively stable, has a higher oxithtion potential than 02 and, being uncharged, can therefore freely cross cell membranes. In addition, H202 in the presence of low concentmtion transition metal, present in almost any solution or in the cells, can produce another ROS, 'OH .Thus, H202 is a good candidate for studying the influence of ROS on cells in vitro. Intracellular 2+ homeostasis in cardiac myocytes is of great functional importance for diastolic resting state ofthe contractile elements, for excitation-contraction-coupling, and for the force of contraction modulated by variation in the magnitude of the transient [Ca211 .Asa result, any [Ca211 changes may influence the heartbeat The relation between ROS and 2+ homeostasis in heart cells is also Significant due to the role ofROS in myocardial ischemia-reperfusion injuiy 2932 These make cardiac cells an important model for the investigation of the LCa2I/ROS relation due to LLL stimulation

Irradiation of mouse spermatozoa by 630 nm HeNe laser was found to enhance calcium transport in these cells. The change in Ca transport was investigated through two approaches, the first employing the fluorescent Ca indicator, Fluo-3 AM and a fluorescence microscopic system, and the second the radiolabeled Ca uptake. In both approaches the effect of light on Ca transport was abrogated in the absence of Ca during the irradiation time, indicating that the effect of light is Ca-dependent. The stimulatory effect of light on Ca uptake was inhibited by treatment with catalase, suggesting H₂O₂ to be involved in light stimulated Ca²⁺ uptake. The stimulatory effect of light on Ca uptake was abolished in the presence of a voltage-dependent Ca-channel inhibitor, nifedipine, indicating the involvement of a plasma membrane, voltage- dependent Ca-channel. In contrast, addition of nifedipine prior to the HeNe laser irradiation did not affect the light-induced rise in intracellular Ca levels, as measured with Fluo-3 loaded sperm cells. Therefore, it can be concluded that this Ca influx occurs via a voltage- insensitive Ca-channel. The stimulatory effect of light on Ca uptake was almost completely abolished by the mitochondrial uncoupler FCCP. These data imply that light affects the mitochondrial Ca transport mechanisms. It is well known that Ca influx from an extracellular environment is an essential component of a signaling cascade leading to fertilization.

The administration of low-level-laserlight for irradiation of the inner ear could represented a new therapeutic model for complex diseases of the inner ear. However, successful therapy requires a well-defined light dosimetry based on a dosimetric analysis of the human cochlea that represents a complex anatomy. The light distribution inside the cochlear windings, produced by an irradiation of the tympanic membrane, was quantitatively measured ex vivo for HeNe laser wavelengths of 593 nm and 633 nm. To obtain the space irradiance within an intact cochlea a correction factor of about 6 has been determined by Monte Carlo calculations. It follows from 3 contributions, first the backscattering of light in the bony parts removed during the preparation procedure of the specimen, second the change of index of refraction from the bony parts to air and third some geometrical factors due to the angular distribution of the radiation. The transmission of light across the tympanic cavity and the promontory depends strongly on the wavelength. Due to the observed spatial intensity variations of a factor 10 and more inside the cochlear windings the optimum external light dose has to be chosen with regard to the tonotopy of the ear.

The aim of contribution is to evaluate the effects of low- level laser radiation on healing process after human molars extraction in lower jaw using frequency 5 Hz, 292 Hz and 9000 Hz. Changes in bone density and monitoring of secretory IgA and albumin levels in saliva were used as a marker of biostimulatory effect. Bone density after extraction and 6 month after surgical treatment was examined using the dental digital radiography. Bone healing was followed by osseointegration of bone structure in extraction wound. Changes of bone density, secretory IgA and albumin levels were compared in groups of patients with laser therapy and control group without laser therapy. Differences in levels of the saliva markers (sIgA and albumin) were found to be significant comparing irradiated and non-irradiated groups, as well as comparing groups irradiated by various modulatory frequencies. Density of alveolar bone (histogram) was examined on five slices acquired from every RVG image. Histograms were evaluated with computer program for microscopic image analysis. Differences of density were verified in area of the whole slice. There were no significant differences found between the bone density in irradiated and non irradiated groups perhaps due to our used therapeutical diagram.

The influence of the low intensive red light on the contracted function of the isolated heart in the post ischemicus period was studied. Sources of the luminous radiation used in the study allow to get the visible light of different ranges and to influence on the local parts of the cardiac muscle.

Most of all renal diseases are accompanied by lowering of kidney functions. That makes the quality of the treatment worse. On an example 69 patients receiving Low-Level Laser Therapy (LLLT), the influence of the laser radiation on a contracting system of blood, on current of an active and inactive tubercular inflammation and on partial functions of kidneys were investigated. Is established, that LLLT does not render influence to a contracting system; promotes stopping of unspecific and moderate peaking of a specific inflammation of kidneys. Is proved, that after a rate of laserotherapy the improving of a blood micricirculation in kidney occurs in 57.9% of patients; a secretion - in 63.1% of the patients; a stimulation of urodynamic is fixed in 79% of cases. Magnification of diuresis, improving filtration and concentration functions of kidneys also is marked.

Exposure of a small skin area (400 cm²) of volunteers to visible incoherent polarized (VIP) light (400 - 2000 nm) in therapeutic doses is accompanied by rapid appearance in the circulating blood of soluble factors able to restore proliferation of X-ray-damaged autologous lymphocytes and to decrease frequency of chromosome breaks. The appearance of a such activity in blood can also be induced without skin irradiation, by in vitro modeling of mixing in the circulation of a small amount of transcutaneously VIP- irradiated blood with the intact blood (one volume of the directly VIP-irradiated blood was added to 10 volumes of the intact blood). Hence, the blood (not the skin) is a major source of the active factors. The data obtained indicate a possibility of release of them from photomodified platelets; moreover, the activity restoring chromosome structure in X- damaged cells has been found in the platelet-derived growth factor and epidermal growth factor (which are known to be associated with platelets) when they were added to physiological concentrations to the culture medium. We assume that activation of cell proliferation based on the effective repair of DNA damaged in situ by endogenous and exogenous factors could be one of the mechanisms of photostimulation of wound healing.

In experiments on 153 non-inbred white female mice lung tumors were induced by subcutaneous injection of urethan (0.5 mg/g) once a week (4 injections as a whole). Mice of the first group were given urethan only. Animals of the second group underwent transcutaneous He-Ne laser irradiation immediately before each urethan injection. Mice of the third group underwent laser irradiation after cessation of 4-week course of urethan. In 2 months after beginning of the experiments, lungs were investigated by light microscopy. It was stated that in animals of the 1st group lung adenomas appeared in 59% of cases. Average amount of tumors per mouse comprised 1.17. In the 2nd group lung tumors were in 82% of animals (p < 0.05). Average amount of tumors per mouse was 1.68. In 2 cases small-cellular lymphocyte-like lung cancer with multicentric growth appeared. Hyperplasia of lymphoid tissue was less in mice with growing tumors. In the animals of the 3d group laser adenomas appeared in 59% of cases. Average amount of tumors per mouse comprised 1.2. It is concluded that He-Ne laser irradiation of animals before injection of chemical carcinogen plays a promising role in the development of murine lung tumors. Laser irradiation after cessation of carcinogen course has no effect on tumor growth.

During same years various medical effect of low-energy laser therapy in veterinary were tested. We established that the laser low-energy therapy can be very effective for treatment such animal's diseases as mastitis and demodekose when certain combinations of laser beam parameters are used. This combinations were taken as the principle of a number of laser veterinary apparatus, which we started to produce at `POLUS'. It is our series of apparatus `VEGA-MB' and `VETLAS-3', which is real used today for dogs and cows treatment in Russia.

We have found that exposure of a small skin area (400 cm²) of healthy volunteers to visible (400 - 2000 nm) incoherent polarized (VIP) light (degree of polarization > 95%) in therapeutic doses (4.8 - 9.6 J/cm²) induces a rapid structural-functional modification of erythrocytes, leukocytes, and some plasma components in the whole circulating volume of blood. We have recorded changes in: lipid peroxidation (LPO) produce content in erythrocyte membranes, deformability and viscosity of erythrocytes, phagocytotic activity of monocytes, cytotoxic activity of natural killer cells against target malignant cells, release of bactericidal proteins by granulocytes, plasma content of LPO-produces and pro-inflammatory cytokines, interleukine-1 beta and tumor necrosis factor-alpha, total anti-oxidant activity of plasma. Most of these effects were of regulative character, as their direction and extent depended on the initial level of the studied parameters: the initially low indices increased, while the initially high ones decreased or remained unaffected. In 24 h the changes were still detectable in 33 - 62% of volunteers. We have shown a great similarity of the blood changes induced by the skin exposure and by the direct irradiation of blood in vitro. Moreover, we obtained an evidence that the light-induced rapid modification of the entire circulating blood resulted from the direct effect upon it of transcutaneously irradiated blood, rather than of other systems of organism. Since many pathological conditions are characterized by an increase in LPO processes, by disturbances in blood rheology, by decrease in natural resistance and immunity, correction of their parameters with the VIP light might be a key mechanism of its therapeutic efficacy.

Irradiation of carp blood leukocytes with a non-laser visible light resulted in a significant inhibition of the spontaneous luminol-dependent chemiluminescence in the cells of a part of the fish. Those leukocytes that were sensitive to the visible light, showed a shorter time-to-peak than the non sensitive, following their stimulation with Ca ionophore. Because a shorter time-to-peak correlates with inflammation, it could be suggested that the visible light susceptible leukocyte reflect a pre-inflammatory state of their donors.