RU2379681C1 - Method of water analysis for biological activity - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention concerns medicine, and can be used for water analysis for biological activity. That is ensured by preparing standard saline of water. Dead-end preparations of nervous cells are prepared and place cells in the saline for preset time. The cells are electrically stimulated by single square-wave pulses of duration 0.1 msec, superthreshold intensity at frequency 1 pulse per hour. Then the excitatory postsynaptic potentials are discharged and recorded both initial and each 2 hours within and after water reaction in cells from a receptor of a-amino-3-hydroxy-5-methylyoxizole-4-propionic acid (AMPA) of the first reciprocal total activating potential of compact neuron group, from a receptor of N-methyl-D-aspartate - the AMPA following reciprocal making active potential of compact neuron group and inhibitory postsynaptic neuron potentials. The biological activity of water in preset time intervals is determined by functioning of nervous cells depending on prolonged water reaction in comparing parametres of nervous cell activity both initial, and within reaction and thereafter.

EFFECT: possibility to compare biological activity of water at molecular-cell level through determining its reaction with dead-end nervous cells.

2 tbl, 2 ex

Description

The invention relates to studies of the properties of water. It is now accepted that water is a biologically neutral substance.

No analogues of the invention were found during the search.

The task is to determine at the molecular-cellular level the effect of water on the functioning of nerve cells, depending on the duration of exposure to water as a biologically active substance.

In modern transplantology, cryopreservation is used for long-term preservation of tissues and organs until they are implanted in the recipient's body. However, this is not acceptable for nerve tissue, since in the process of freezing / thawing, the membranes of nerve cells and their functional contacts, synapses, are destroyed. The urgent problem of ensuring the viability of the nervous tissue, its functioning.

The essence of the proposed method for determining biological activity consists in the preparation of standard saline solution from water, the preparation of surviving preparations of nerve cells, placement of cells for a predetermined time of 11 hours in saline solution, cell stimulation with single, rectangular electric pulses of 0.1 ms duration, suprathreshold intensity, with a frequency of 1 pulse per hour. Allocation and registration of the initial and every 2 hours during and after exposure to water the occurrence of inhibitory postsynaptic potential

(TP SPm), the exciting postsynaptic potential slow in cells from the AMPA receptor - (α-amino-3-hydroxy-5-methylioxazole-4-propionic acid) - the first response total activating potential of the compact group of neurons and from the NMDA receptor - (N- methyl-D-aspartate) - the subsequent activating potential of a group of neurons after AMPA, and from the action potential (PD), the determination of the biological active effect of water when comparing the parameters of the induced activity of the initial nerve cells during the incubation process and after its completion at predetermined intervals, cells functioning in the water depending on prolonged (11 hours) exposure to water.

The method for determining the biological activity of water is as follows.

Prepare a standard saline solution from water. Preparing preparations of surviving nerve cells in the form of slices of the nerve tissue of the brain or single, prepared, or in tissue culture. These nerve cells for a given time of 11 hours (determined by the limitation of the time of work according to the technical passport of the electrophysiological installation) are placed in saline. Cells are stimulated with single rectangular electric pulses with a duration of 0.1 ms, above threshold intensity with a frequency of 1 pulse per 1 hour. Allocate the potentials PD, AMPA VPSP, NMDA VPSP, TPSPm initial and every 2 hours in the process of being in water.

Registration time is stipulated for the convenience of comparing the activity of water from various sources or before and after any industrial water treatment. The activity of biological water is determined by comparing the parameters of the induced bioelectric activity of the nerve cells of the original and during incubation in water at predetermined time intervals during their 11-hour stay in the test water.

Example 1

The biological activity of local tap water was determined by its effect on the functionality of nerve cells in sections of brain nerve tissue according to the parameters of their induced bioelectric activity, depending on the duration of exposure to water. Cells were incubated in water for 11 hours, see table 1.

Table 1 Parameters potentials Evoked bioelectric activity of neurons (in μV) and duration of exposure to water (in hours) Initial activity (μV) Activity after 1 h (μV) Activity after 3 hours (μV) Activity after 5 hours (μV) Activity after 7 h (μV) Activity after 9 hours (μV) Activity after 11 h (μV) PD 401 391 387 385 386 380 378 AMPA VPSP 378 375 372 370 285 280 274 NMDA UPU 82 89 80 67 55 23 fourteen TPSPm 32 29th 17 12 8 3 2

As can be seen from the data presented in table 1, when exposed to distilled local tap water during a long (11 h) incubation of brain sections, the functioning of the nerve cells of the sections decreased in potentials: PD by 6%, AMPA EPSP by 18% compared to the initial values. The activity of the most sensitive components of potentials - NMDA of EPSP and TPPSm - decreased most significantly. Thus, the amplitude at the NMDA receptor of EPSP decreased by 83% compared to the initial one. Even more significant was a decrease in the activity of inhibitory mechanisms, determined by the amplitude of the TPPSm potential, by 94%. Thus, prolonged incubation of sections in standard saline solution prepared with tap distilled water reduces the functionality of nerve cells.

Example 2

Determination of the effect of saline from the same tap water, but processed in the Water Treatment Device (application for an invention dated 30.10.2007 No. 2007140226/17), on brain nerve cells by the parameters of the induced bioelectric activity. The characteristics of the incubation test solution (concentration of salt components of pH, osmolarity) was identical to the control. Only the tested water is changed.

table 2 Parameters potentials Evoked bioelectric activity of neurons (in μV) and duration of exposure to water (in hours) Initial activity (μV) Activity after 1 h (μV) Activity after 3 hours (μV) Activity after 5 hours (μV) Activity after 7 h (μV) Activity after 9 hours (μV) Activity after 11 h (μV) PD 390 398 381 38Q 382 385 381 AMPA VPSP 357 371 385 383 384 386 385 NMDA UPU 76 81 85 74 75 73 74 TPSPm 25 29th 33 23 24 23 24

As can be seen from the data presented in table 2, a prolonged (11 h) exposure to purified nerve tissue by purified water supports the stable functioning of nerve cells. Before the level of functioning is stabilized, the process of cell activation develops after 1-5 hours of incubation. These data indicate the functionality of nerve cells during prolonged storage in solution in purified water. Since all the characteristics of the incubation solution were the same as in the first example, with the exception of the water itself, which was purified in the device, then, therefore, these effects are due to changes in the properties of water. Such a long-term (11 hours) stable (see Table 2) functioning of nerve cells can be explained by the fact that this water nourishes the cells with energy throughout the entire process of incubation in it.

The proposed method allows to determine the biological activity of water at the molecular-cellular level by determining its effect on surviving nerve cells exposed to the test water for a given time. The method allows to determine and compare the level of functionality of cells at predetermined points in time depending on the duration of their stay in the test water.

The method shows that water is a biologically active substance with different levels of activity in different sources. "A device for water purification" (application for invention No. 2007122226/17) was submitted for research to the Institute of Physiology named after I.P. Pavlov of the Russian Academy of Sciences by the authors Yu.A. Levchenko and Yu.I. Sviridov.

Claims (1)

A method for determining the biological activity of water, which consists in preparing a standard saline solution from water, preparing surviving nerve cell preparations, placing cells for a predetermined time in saline solution, electrical stimulation of cells with single rectangular pulses of 0.1 ms duration, suprathreshold intensity with a frequency of 1 pulse per hour, lead and registration of the initial and every 2 hours during and after exposure to water excitatory postsynaptic potentials arising in the cells from the receptor α-amino-3-guide roxy-5-methylisoxazole-4-propionic acid (AMPA) of the first response total activating potential of a compact group of neurons, from the N-methyl-D-aspartate receptor - subsequent after AMPA response activating potential of a compact group of neurons and inhibitory postsynaptic potentials of neurons; and determining the biological activity of water at predetermined time intervals for the functioning of nerve cells, depending on the prolonged exposure to water by comparing the parameters of the activity of nerve cells, both initial and in the process of exposure and after its completion.