Method for assessing artificial light exposure on factors of peripheral blood and congenital immunity with use of laboratory animal models

FIELD: medicine.

SUBSTANCE: senior guinea pigs are exposed to optical light generated by light emitting diodes or fluorescent tubes at a colour temperature of 4,500 K within the wavelength region of 360-460 nm for various time distances. The exposure is assessed by functional activity indices of neutrophilic granulocytes and a mononuclear cell count.

EFFECT: using the method requires no expensive equipment, critical chemicals; it is of a particular importance for evaluating the biological safety of new artificial light sources embedded into a light-colour medium; it provides expanded information on biological effects of optical light.

3 tbl

 

The invention relates to experimental medicine, in particular to assess the impact of artificial lighting on the condition and functional activity of factors of peripheral blood neutrophils and mononuclear cells of Mature Guinea pigs sensitive to the spectral characteristics of the generated fluorescent and led lamps with a color temperature of 4500 K in the wavelength range 360-460 nm for various time intervals and can be used to assess the impact of artificial light optical range generated by the LEDs and fluorescent lamps on a number of important system parameters of homeostasis sensitive laboratory animals, namely, the functional activity of phagocytes and peripheral blood mononuclear cells using laboratory animal models, since under the action of artificial light with a color temperature of 4500 K in the wavelength range 360-460 nm generated by fluorescent lamps in laboratory animals in vivo recorded the change in the functional activity of neutrophilic granulocytes and the number of mononuclear cells, including marked increase in the biocidal capabilities of neutrophils detected in nitroblue tetrazolium (NBT-test) 26%, increased number of mononuclear cells in the peripheral blood of laboratory animals �and 20%. Under the action of led light sources with similar spectral characteristics not observed changes in the functional activity of peripheral blood of Guinea pigs.

To date, to assess the impact of light generated by artificial sources used in the analysis of the functions of the visual analyzer, which allows us to offer the solution of a number of hygienic standardization problems associated with the use of energy efficient lighting [M. E. Gusel'nikov, 2008; Kuchma V. R. et al, 2013]. With the discovery of a new type of photoreceptor in the eye and photoacceptors of neutrophils appeared understanding of non-visual biological effects of light on the body, what is happening with the participation of the hormones melanin and melanopsin [Mahmuthodjaev A. S., 1999; Brainard, G. K. et al, 2008]. When hit by light in cell-receptors starts a chemical reaction with the participation of photopigment melanopsin, which results in the production of electrical impulses. In the retina, light waves of a certain wavelength converted into energy nerve impulse that is transmitted along the optic nerve to the upper part of the spinal cord and in the occipital lobe of the brain, affecting the main control centers of the body located in the brain, including the activity of the innate im�unnuh mechanisms of regulation, by the neutrophils - cells that have receptors for melatonin and is specific to the perception of a quantum of light [Alenichev I. F. et al, 2006; Dolgushin I. I., 2001; Gisinger O. A. et al, 2013]. Proposed in 2013 method of evaluation of the effects of light generated by the led sources on the function of neutrophilic granulocytes in the peripheral blood can be used to assess the impact of the light generated by led and other artificial sources on cellular factors of innate immunity [Gisinger O. A. et al, 2013]. The proposed method has some drawbacks associated with a narrow spectral band of sensitivity of neutrophils, labor-intensive technologies of separation and maintaining cell cultures requiring the presence of chemicals imported. In addition, neutrophils are not the entire population of blood cells, and mononuclear cells accounts for 30-40% of formed elements, which makes the study of their population dynamics and relevant when considering the possible biological effects of artificial light.

Used today as a monomethod method of assessing the effect of light on the body on the assessment of the response of the visual analyzer man has several disadvantages [Kuchma V. R. et al., 2013]. In particular, the main drawback IU�ode is the use of different age groups of human population, especially children aged 5 to 7 years as the objects of study, the difficulty in obtaining voluntary informed consent from all participants, including minors, decisions, possible discomfort to study participants younger age groups from the effects of artificial light generated by led and fluorescent lamps, and its subjective assessment in subjects in the experiment reduces the validity, convergence and reproducibility of results.

V. N. Anisimov proposed evaluation of the effects of artificial light by analyzing the biochemical parameters, enabling the identification of endocrine disorders, resulting from prolonged exposure to artificial light [Anisimov V. N., 2008]. The disadvantage of this method is its low sensitivity and specificity arising from possible fluctuations in biochemical parameters as a result of not only exogenous light, but also the possible endogenous reactions of the organism, the complexity of the selection criteria for inclusion and exclusion of subjects from the survey, the high cost of chemicals, human labor costs.

B. C. Kryshtal as one of the methods of studying the action of light has offered to conduct an analysis of mental health and�asemota, light and color perception with the use of psychological testing. The disadvantage of this method is the use of human rights as an object of study arising from this subjective evaluation of the action of light and unilateral approach to analysis, allowing you to install only the mental state of the subject at the time of the study, not taking into account indicators neuroimmunoendocrine regulation [Kryshtal B. C., 2005]. However, the values of individual parameters obtained from the results of testing, questionnaires or complex testing + surveys do not always reflect the true state of the light influence on all organs and systems of the person, connected with a variety of accompanying systemic dysfunction disorders.

Presents literature data on the prediction of the risk of occurrence of the pathology of the cardiovascular system under the action of light generated by LEDs or other artificial light sources, to assess the degree of heart rate variability and the analysis of vegetative indicator rhythm of the heart [Kudryashov E. A., 2008]. The use of this method can be extremely popular in medical hospitals, equipped with modern energy-efficient artificial devices, however, is of limited use, due to complex and expensive about�of equipment and training of personnel and the possibility of its use for carrying out of sanitary-epidemiological control Biosafety action of artificial light.

There are methods using questionnaires to assess the impact of light on the psychosomatic state of the patient, but their use is also not excluded the influence of subjective endogenous and exogenous factors (features of chronotype, health status at the time of the study) affecting the results.

Since the values obtained by these methods research may contain inaccuracies, for example by questioning or analysis of blood biochemical parameters, the most accurate reflection of the influence of the light generated by the led media can be a study of the effect of light on the model cells responsive to various exogenous effects and having photoreceptors for light quanta - neutrophil granulocytes [Gisinger O. A., 2013, the application for invention No. 2013142730 from 19.09.2013, the invention application No. 2013158668 from 26.12.2013] subject presented in the abstract of the shortcomings of the method which can be adopted as a basis and taken as a prototype.

The basis of the invention consists in the creation of experimental models in laboratory animals - Guinea pigs are sensitive to the studied spectral characteristics, making possible further statistically analyze the effects of light, GE�erasemode led or other artificial sources on the condition and functional activity of the cellular elements of peripheral blood Mature Guinea pigs.

This problem is solved due to the fact that in the proposed method, the biological evaluation of the light generated by the led or other artificial light sources, comes with complete clinical and functional results, excluding the direct participation of the (people) of different age groups as models for the experiment. Application of the method does not require expensive equipment, scarce chemicals available in the practice of research laboratories, with facilities for keeping laboratory animals, particularly important in the evaluation of new Biosafety introduced in environment colour-light therapeutic educational and other social facilities sources, expands information on biological effects of light in the optical range.

The claimed method allows to objectively evaluate the effects of physical factors on the factors of homeostasis of an organism using models sensitive laboratory animals. Previously, such analysis could be undertaken using analysis of isolated cell cultures of neutrophil granulocytes to assess the impact of low-intensity laser radiation on neutrophils of peripheral blood of donors and secrets of the urogenital tract in the experiment [Gisinger O. A. et al., 2009; 2013], without consideration of the response, which will�and mononuclear cells.

The authors first proposed to use this method for studies of the biological effects of light, optical range generated by led and other artificial sources.

For solving the above problems were investigated in three experimental models in laboratory animals (Guinea pigs). The use of these animals is determined by their sensitivity to the action of light in the optical range. Depending on the choice of light source in the first model in laboratory animals acted by natural light in normal conditions chronogram (natural alternation of day and night),control group), T=4500 K (white light), the wavelength of 360 nm; in the second model in laboratory animals operated light generated by fluorescent lamps T=4500 K (white light), Dina wave 360 nm; in the third model, operated by the light generated by the led sources T=4500 K (white light), the wavelength of 360 nm. The study took into account the composition, absolute and relative content of formed elements in the peripheral blood of laboratory animals, their functional activity. To conduct the study groups were formed: group 1 (control) animals were housed under natural light; group 2 - on laboratory animals affected by light generated by lumines�entname lamps; group 3 - on laboratory animals affected by light generated by the LEDs. Points of monitoring indicators have been 10 days, 20 days of the experiment. The experimental and control groups of animals during exposure to artificial light was specially equipped for the experiment areas. Applicable light field, the light of the performance of illuminance, it is configured so that at any point in the study of the deviation of the surface density of the luminous flux was lower than 10% from the specified parameters. The obtained results were subjected to statistical calculation of the arithmetic mean and its standard error. On the reliability of differences of average values was judged using nonparametric Mann-Whitney test. The differences were considered significant at p≤0.05.

Qualitative analysis showed that the ten-day exposure to light generated by natural light sources in laboratory animals in natural chronotype does not change the absolute and relative number of cellular factors in the peripheral blood and their functional activity. The quantitative composition of leukocytes in the peripheral blood of laboratory animals under led and fluorescent light sources within the 10 days provided no�Wii animal of DS showed the presence of significant differences in the content of formed elements: reduction stab granulocytes 26.6%, in animals under the influence of fluorescent lamps, the reduction of lymphocytes 10% in animals under the influence of led lamps. The results of the influence of different light sources on the composition and functional activity of cellular factors in the peripheral blood of Guinea pigs in vivo (10 days exposure to artificial light) are presented in table 1.

The quantitative composition of leukocytes in the peripheral blood of laboratory animals under led and fluorescent light sources within 20 days showed no changes in absolute content and content band neutrophils, eosinophils, and lymphocytes in the groups of animals under natural conditions (control) and led lighting (p1>0,05). Absolute and relative number of segmented neutrophils in the peripheral blood of laboratory animals under the light of fluorescent lamps was increased compared with the figures of animals under the action of led lighting and natural light. Absolute monocytes in the group animals in led lighting (5,63±0,41%) statistically significantly different from the levels of monocytes in the stomach�s from the control group (7,50±0,79%) and performance in animals under fluorescent lighting (7,75±2,42%), and relative indicators of monocytes in animals under the influence of fluorescent lamps in 3 times higher than the values of control group and groups of animals, located in led lighting (p1<0.05, p2<0,05). The results of the influence of different light sources on cellular factors peripheral blood in vivo (20 days exposure to artificial light) are presented in table 2.

Data analysis functional activity of the cellular elements of peripheral blood of Guinea pigs under the conditions of artificial lighting within 10 days revealed differences in phagocytic activity of neutrophil granulocytes in animals under natural light, and in animals under the action of fluorescent and led lamps (p1,2<0,05). Found that after exposure to light of fluorescent lamps in the blood of the examined animals had increased the number of active neutrophils in spontaneous NBT-test in comparison with natural lighting, and led lighting, increased their phagocytosis. The results indicate a pronounced stimulating effect of the light generated by the fluorescent lamp, while the lamp NAC�Libanius and led carriers to generate light, does not cause pronounced immunological changes in the system of neutrophilic granulocytes in the peripheral blood of laboratory animals. The results are presented in table 3.

Thus, the influence of the light generated by fluorescent and led lamps, the light within the temperature 4500 K, the wavelength of 360 nm for 10-20 days leads to a statistically significant change in the number and functional activity of the cellular elements of peripheral blood of laboratory animals (Guinea pigs) compared to natural light, and the influence of the light of fluorescent lamps leads to significant activation of oxygen-dependent metabolism and phagocytic activity of neutrophil granulocytes in the peripheral blood of Guinea pigs in vivo.

The application of the method to assess the impact of artificial lighting on the condition and functional activity of factors in the peripheral blood by exposure to sexually Mature Guinea pigs, allows to solve the problem laboratory monitoring of efficacy and Biosafety of artificial sources of light generated by fluorescent lamps and LEDs with attraction as the model is sensitive to the spectral characteristics of the studied laboratory animals.

Offer FPIC�b differs from the existing ones, using laboratory animals (Guinea pigs) having a photosensitive receptors to study spectral characteristics of artificial light is acquired the opportunity to more efficiently, without the use of humans as subjects in research, in vivo to evaluate the biological effects of artificial light sources on the status of the basic parameters of homeostasis of the microorganism, thereby to facilitate the prediction and elimination of risks that arise when planning colour-light of the human environment and the surrounding biological objects. Due to the correctness of the method (avoiding the need to use human rights as an object of study), compliance with the rules of the treatment of animals, ease of handling and inexpensive laboratory methods the method of assessment available to the majority of hygiene, immunological laboratories, research institutes dealing with hygienic standardization of artificial light and has permission to work with laboratory animals.

The inventive method may find wide application in immunology, the study of immunotropic or immunosuppressive effects when conducting Santana-hygienic evaluation of residential and light industrial facilities, cosmetically on its compliance with the criterion of "industrial applicability".

Literature

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2. Brainard G. K. the Perception of light as a stimulus for non-visual reactions / G. K. Brainard, I. Provencio // Svetotekhnika.- 2008. - No. 1. - S. 6-12.

3. Bike K. Subjective assessment of color depending on the range of light sources / K. Biske, K. Vandal, K. Jungnitz // Svetotekhnika.- 2007.- No. 5. - S. 14-17.

4. Gisinger O. A. Influence of low-intensity laser radiation on neutrophils peripheral blood donors in the experiment. / O. A. Gisinger, K. G. Ispahan, Kolesnikov O. L. // Immunology. - 2009. - Vol. 30, No. 5. - P. 263-267.

5. Gisinger O. A. Research approaches in the field of security lighting in megapolis / O. A. Guisinger, M. V. Usikov, O. R. Bokova, etc. / / Semiconductor lighting technology. - 2013. - Vol. 1, No. 21. Pp. 60-61.

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A method of assessing the impact of artificial lighting on the condition and functional activity of factors in the peripheral blood by exposure to sexually Mature Guinea pigs as animals, are sensitive to the studied spectral characteristics of the radiation of the optical range generated by LEDs or fluorescent lamps with a color temperature of 4500 K in the wavelength range 360-460 nm for various time intervals, wherein the impact assessment produced by the indicators of the functional activity of neutrophilic granulocytes in the peripheral blood and the number of mononuclear�x cells.



 

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15 cl, 9 dwg

FIELD: medicine.

SUBSTANCE: clinical research aimed at detection arterial hypertension is performed. Echocardiographic examination with the detection of signs of sclerogenic changes of the aorta is carried out. A value of function F is calculated. 2 demographic parameters - sex and age are taken into account, 1 clinical parameter is arterial hypertension and 1 echocardiographic parameter - presence of the sclerogenic injury of the aorta. Then the value of a probability level P is calculated. Patients with unchanged coronary arteries have the value P lower or equal 0.204. In patients with non-obstructive coronary-sclerosis it is over 0.204.

EFFECT: method makes it possible to exactly, at early stages of the disease, carry out non-invasive diagnostics, determine indications for effective treatment actions, ensures the prevention of complications due to the application of available clinical and instrumental methods, including in calculation most significant clinical signs, application of methods of determining the probability of the presence of non-obstructive coronary-sclerosis.

2 dwg, 2 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: blood pressure is measured according to the standard procedure on the brachial artery by means of the Riva Rocci Korotkoff sphygmomanometer. The vegetative status is assessed by the rest heart rate variability parameters. The endothelial vasomotor function is described as a percentage measure of a resistance index after inhalation of Salbutamol 400 mcg, an endothelium-dependent vasodilator as related to a basal resistance index by photoplethysmography. The derived data are used to determine an adequate basal pulse wave velocity and a pulse wave velocity following the sublingual administration of nitroglycerin 500 mcg, for each patient individually by original formulas.

EFFECT: method enables providing the more accurate determination of the adequate basal pulse wave velocities and the pulse wave velocity following the administration of nitroglycerin taking into account the vegetative status.

2 dwg, 1 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine, namely to organising and conducting insulin therapy for a patient. Determining a user's dose of basal insulin is ensured by using a device for insulin administration and a portable device for diabetic data control. The method involves measuring the number of the user's blood glucose concentrations over the number of time intervals. The collected data are used to state if the user has performed the minimum number of measurements of the fasting blood glucose concentration over at least one of the four time intervals prescribed. That is followed by stating if the collected data indicate one of the first schedule showing the low blood glucose concentration, and the second schedule showing the lower blood glucose concentration than in the first schedule showing the low glucose concentration. Thereafter, the method provides considering if the basal insulin dose correction in the portable device for the first and second schedules correspond to each other; the user is warned, and the basal insulin dose correction is prohibited in any of the devices. Taking preventive measures aiming at measuring the basal insulin dose by means of the devices involves collecting the data, and the preventive measures are taken to avoid the user's hypoglycaemic state.

EFFECT: group of inventions enables rendering aid in the patients suffering from diabetes by means of the devices for determining the basal insulin dose and controlling the blood dose level in an insulin dose and taking the preventive measures.

27 cl, 11 dwg

FIELD: medicine.

SUBSTANCE: method involves carrying out ultrasonic scanning examination of subclavian artery over its whole extent in physiological arm position with arterial blood pressure being measured in the middle one third of the arm. Next, when applying compression tests, blood circulation parameters variations are recorded in distal segment of the subclavian artery with arterial blood pressure being concurrently measured. Three degrees of superior thorax aperture syndrome severity are diagnosed depending on reduction of linear blood circulation velocity and arterial blood pressure compared to their initial values. Mild one takes place when linear blood circulation velocity reduction reaches 40% and arterial blood pressure 20% of initial level, moderate one when linear blood circulation velocity reduction reaches 70% and arterial blood pressure 50% and heavy one when linear blood circulation velocity reduction is greater than 70% of initial level and arterial blood pressure is greater than 50% to the extent of no blood circulation manifestation being observed in the subclavian artery.

EFFECT: high accuracy of diagnosis.

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