Method for determining substances transport intensity changes in prevailing directions between blood and non-mineralized organs

FIELD: medicine.

SUBSTANCE: method involves introducing radioisotope to animals and further repeatedly determining radioisotope inclusions percent in blood and in and in non-mineralized organs in given time intervals, calculating relative radio-activity RRA for an animal examined at each time as ratio between radioisotope inclusions percent in non-mineralized organs to radioisotope inclusions percent in blood. Then transport intensity is determined in prevailing directions in each examination time by applying RRA difference factor (DF_RRA), calculated as difference between the subsequent and previous RRA values. The received values of DF_RRA factor changes are interpreted in terms of intensity time fluctuations and radioisotope transport direction for each organ.

EFFECT: enhanced effectiveness in determining intensity dynamics and prevailing direction of substance transport between non-mineralized organ and blood.

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The invention relates to the field of biology and medicine, can be applied biochemists, physiologists, pathophysiology, experts in laboratory diagnosis, long-term space flights in the state of weightlessness and clinicians.

There is a method of determining the direction of transport and metabolic rate between mineralized tissue and contacting the biological fluid, characterized in that the injected radioisotope one pair of experimental animals receiving standard feed, and the second pair of experimental animals treated with sucrose cariogenic diet, determine the % include radioisotopes in mineralized tissue and contacting the biological fluid in a period of 24 h, calculate the relative radioactivity of PR for each animal ORA₁, ORA₂, ORA₃and ORA₄by the formula

and when the values of PR>1, draw conclusions about the direction of transport of a substance from a biological fluid in mineralized tissue; ORA<1, draw conclusions about the direction of transport of the radioisotope in the opposite direction, and when the PR 1.0, conclude that both directions are balanced among themselves; then for these groups of experimental animals calculates the intensity of transport in the prevailing direction is the situation using the ratio of the difference ORA CU _ORAformula CU_OR=ORA₂ORA₁and CU_OR=ORA₄ORA₃; CU_ORAgreater than zero indicate a +, CU_ORAless than zero indicate the sign; if the values compare CU_ORAhave opposite signs, then the higher the intensity of the radioisotope transport of the biological fluid in mineralized tissue will be at CU_ORAwith a large value regardless of the sign (patent No. 2242007 of the Russian Federation. The method of determining the direction of transport and the state of the metabolism between mineralized tissue and contacting the biological fluid. Petrovich Y.A., roadside R.P., Kitsenko S.M. /Mosk. state medical-dental. University, Publ. 2004, No. 34).

However, when performing the methods of the prototype only once compared the intensity and direction of transport of radioactive substances in the prevailing direction between the biological fluid and mineralized tissue after 3 and 24 h for [³⁵Se]methionine or only after 6 and 24 h for [⁷⁵Se]selenate after injection of a radioactive substance. Using the prototype method is not detected, what happens to the radioisotope before and after the specified time, as well as between the two definitions. Since the tissue of bones and teeth, containing up to 60-90% of mineral components from their mass, hundreds and t is thousands of times different from remineralizing bodies high specificity of the chemical composition, metabolism, permeability, experiments with the definition of the transport of substances in the prevailing direction between the blood and saline bodies cannot testify about what is happening with the transport of substances between the blood and demineralizovannykh bodies. In addition, a single definition proposed in the method prototype is generally impossible to identify possible oppositely directed changes in transport of substances in the prevailing directions between the blood and the bodies. This requires multiple sequential tests for a long time.

However, knowledge of possible differences in the intensity of the transport of substances in the prevailing directions between the blood and bodies, with multiple consecutive definitions may submit to the experimenters completely new information about the patterns of transport and metabolism, and clinicians may identify new approaches to diagnosis and treatment of various diseases.

In addition, the detection time of the maximum and "targeted" delivery of specific substances, including therapeutic, in a specific organ (or organs) for a long time, and not just within one day, may be important for diagnosis and therapy.

The vast majority of practically available to study the connections in the state β emitters³H,¹⁴C,³²P low energy radiation is completely absorbed by soft tissue thickness up to 1-3 mm, and only a few available connections marked γ-emitters radiation which penetrates from the deep layers through tissue located more superficially. In addition, γscan cannot replace our proposed method because it determines the total radiation of different organs and blood, and not separately for blood and organ, as in our method.

Object of the invention is the determination of the difference in intensity of the transport of substances in the prevailing directions between the blood and the bodies that should improve the objectivity of research due to multiple definitions in the blood and in the organs of radioactive or non-radioactive substances for a long time after its introduction experimental animals.

This is achieved by the fact that the group of animals once intraperitoneally injected radioisotope, after different times of rats consistently derive from experience, calculate the % include label in the blood and demineralization organ (e.g. liver or kidney), calculate the relative radioactivity (ORA) blood/demineralizovannykh body by formula 1:

,

then calculate the intensity of the transport side is dominant directions using the ratio of the difference of PR (CU _ORAthe formula 2: KP_OR=ORA₂ORA₁, CU_OR=ORA₃ORA₂, CU_OR=ORA₄ORA₃etc.; CU_ORAabove the zero line when the numeric value of the following ORA greater than the previous one, in a given period of time between their definitions is dominated by the transport of the radioisotope from the blood into the body, or when the numerical value of the following ORA lower than the previous one, is dominated by the transport of the radioisotope in the opposite direction; CU_ORAbelow the zero line when the numeric value of the following ORA is still lower than the previous one, is dominated by the transport of the radioisotope from the body in the blood, or when the numerical value of the following ORA higher than the previous one, is dominated by the transport of the radioisotope in the opposite direction; a greattraffic radioisotopewill be at a higher difference values of the two closest CU_ORA; the difference in the intensity of transport is calculated when comparing the values of the two nearest CU_ORAwith the same signs (+or -)by subtracting the value of the smaller CU_ORAfrom a larger CU_ORA; when comparing the values of two CU_ORAwith different signs summarize their values; similarly to the way youapplied in the study of traffic between the blood and mineralizovannyh and bodies, as well as in observations with non-radioactive substances.

Presents a new method allows to determine the difference in the intensity and rhythm fluctuations of the transport flows of substances in the prevailing directions between the blood and the body and quantitatively to characterize him.

The method is as follows: group of animals once intraperitoneally injected radioisotope, after different times of rats consistently derive from experience, calculate the % include label in the blood and demineralization organ (e.g. liver or kidney), calculate the relative radioactivity (ORA) blood/demineralizovannykh body by formula 1:

then calculate the intensity of transport in the prevailing direction using the ratio of the difference of PR (CU_ORAthe formula 2: CU_OR=ORA₂ORA₁, CU_OR=ORA₃ORA₂, CU_OR=ORA₄ORA₃and so on; then for each term of study determines the rate of transport in the prevailing direction using the ratio of the difference of PR (CU_ORA), calculated as the difference between the next and previous values of PR. Change the obtained values of the coefficient of CU_ORAdetermine temporal variations of intensity and direction of transport of the radioisotope is for every body.

Depending on the objectives of the study can increase the number of studied organs, methods of analysis and animal per period of time, and also change the duration and the number of intervals between definitions.

Research % inclusion, ORA, CU_ORAand differences in the intensity of transport after injection of [⁷⁵Se]selenate shown in examples 1, 2 with the liver and examples 3, 4 kidney, and figure 1 of [⁷⁵Se]selenate was administered to rats one month and three months of age intraperitoneally based 20000 counts/min per 1 g of the weight of the animal.

Example 1. Changes fluctuations CU_ORAliver/blood in rats months of age (table 1, figure 1) had the appearance of three-hump curve. CU_ORAfrom inception to 1 h was +7,5, from 1 to 3 h decreased to +2, from 3 to 6 h increased slightly to almost +2,5, 6 to 12 h plummeted to -5,6, from 12 to 24 h returned to subprojectname value +0,36, from 24 to 48 h again fell sharply below -4, 48 h and 192 h increased to -0,78.

Example 2 (table 1, figure 2). The three-month rats fluctuations CU_ORAliver/blood were two humps. From inception to 1 h CU_ORAdramatically upgraded to +9, with 1 to 3 h and 3 to 6 h significantly reduced almost to -7, in the period from 6 to 12 h increased to +4, and gradually reduced to -1 to the end of the experiment.

Example 3 (table 1, figure 1). At the month-old rats curve fluctuations CU_ORAkidney/blood also what had been the three humps. From 0 to 3 h values was +3,53 - +3,39, from 3 to 6 h sharp decline to -1,30, from 6 to 12 h large increase to +2,2, and from 12 to 24 hours more a sharp decline to -4,4, from 24 to 48 h again increased to +2 and 48 h and up to 192 h again a significant reduction to -2,76.

Example 4 (table 1, figure 1). Two-humped fluctuations CU_ORAkidney/blood from a three-month rats also had the opposite directional variations, but with a different rhythm of transport. To 1 h after injection⁷⁵Se CU_ORAabove +3,0, then from 1 to 3 h moderate decrease to +1.5, and a sharp decrease to -4 from 3 to 6 hours, alternating significant rise to +5,5 6 to 12 hours, then 12 hours until the end of the experiment, a gradual decrease to almost-2. Changes CU_ORAin the three-month kidneys of rats with 3-6 h before 24-48 h differed from the month of rats at a much slower rate fluctuations.

Three-hump curve in the liver and kidneys monthly rats and two in the three-month rats, probably due to more intensive metabolism in young will vozraste than adult rats.

Figure 2 is mapped fluctuations age-related changes in differences in the intensity of transport [⁷⁵Se]selenate, first, between the blood and the liver (A), secondly, between the blood and kidneys (B). The three-month rats in the first half experience a slight increase in the differences in the intensity of transport between the blood and the liver; at the month-old rats in the first half of the experience noted the correspondingly sharp decline, then a significant upturn, and then in two age groups, there is a decrease until the end of the experience. In the kidney in two age groups in the first half of experience has increased differences in the intensity of transport, in the second half of the experience - reduction, but more moderate than in the liver.

Figure 3 is mapped fluctuations changes differences in the intensity of the transport of radioactive isotope between the blood and the liver or blood and kidneys at the month-old animals (A), during the whole experience, except for the last interval. They are oppositely directed. The three-month rats (B), on the contrary, in the early and mid experience changes were parallel in the form of lifting and the next lower, and only at the last time they were the opposite.

Figure 4 shows % inclusion in the serum and in demineralization body, ORA and CU_ORA. Clearly visible fluctuations CU_ORAstarting with 0-1 h and ending 48-192 h after injection, indicated by thin arrows. Bold arrow 4 additionally shows a possible example of the fluctuations of the transport of substances. When the source is CU_ORAabove the zero line and the greater value of the next CU_ORAthan the previous one , is dominated by the transport of the radioisotope from the blood into mineralized tissue (A); when the source is CU_ORAabove zero is the turn and a smaller value of the next CU _ORAthan the previous one , is dominated by the transport in the opposite direction (G); if the source is CU_ORAbelow the zero line and a smaller value of the next CU_ORAthan the previous one , is dominated by the transport of the radioisotope of mineralized tissue in the blood (B); when the source is CU_ORAbelow zero and a larger value of the following CU_ORAthan the previous one , is dominated by the transport of the radioisotope from the blood into mineralized tissue (At).

The proposed method for determining the dynamics of changes in the intensity of transport of substances in the prevailing directions between the blood and demineralizovannykh bodies has a high degree of objectivity that allows it to be applied in the experiment and in the clinic to examine the possibility of a "targeted" delivery of radioactive substances or stable substances, which did not exist before in the body, the body for the purpose of diagnosis and treatment. So, for example, quantify the temporal dynamics of PR and CR_ORAyou can apply when selecting a radioisotope for the differential diagnosis of malignant and benign tumors, when malignant neoplasms and metastases accumulate more radioactive than benign tumors and healthy bodies. Obviously, more suitable for diagnosis and treatment will be a radioisotope with Baltimor _ORAdominated by transport from the blood into the body.

Thus, in the study of CU_ORAand differences in the intensity of transport in the prevailing directions between the blood and organs of rats found when comparing the nearest CU_ORAage differences in their amplitude and frequency.

Based on the obtained fundamental information on the physical law of interference fluctuations of different types of waves, it is possible to perform correction of the deficiency of a substance at a specific time in a specific organ with additional introduction of substances so that the maximum concentration after repeated administration of the substance was at the time of its minimum level in the tissue prior to the introduction.

The way to determine the dynamics of changes in the intensity of transport of substances in the prevailing directions between the blood and demineralizovannykh bodies, characterized by the fact that animals injected radioisotope, then repeatedly at set intervals to determine percent incorporation of the radioisotope in the blood and in demineralization body, calculate the relative radioactivity of PR for the studied each term animal as the ratio of the % inclusion of the radioisotope in demineralization authority to % inclusion for the sake of the isotope in the blood, then for each term of study determines the rate of transport in the prevailing direction using the ratio of the difference of PR (CU_ORA), calculated as the difference between the next and previous values of PR, and change the obtained values of the coefficient of CU_ORAdetermine temporal variations of intensity and direction of transport of the radioisotope for each organ.