RU2220420C2 - Method for detecting glucose utilization by tissues - Google Patents

Abstract

FIELD: medicine, laboratory diagnostics. SUBSTANCE: one should detect the rate of glucose decrease, moreover, one should apply 0.02 ml 8%-glucose solution onto presterilized (due to dry heat at 120-140 C) test-carrier, place glucose test-carrier (GTC) into superior labial-gingival fold after its 4-fold purification with a tongue, after 10-min-long exposure one should place a paper part of GTC into a tube with 1.0 ml distilled water by shaking it constantly and detect glucose concentration in aqueous glucose solution obtained 10 min later, and according to glucose decrease value against initial quantity one should detect the coefficient for glucose absorption by tissues. EFFECT: higher efficiency of detection. 2 ex

Description

 The invention relates to medicine, namely to identify a predisposition to diabetes, its latent course, as well as other disorders of carbohydrate metabolism.

 It is known that a violation of the ability of tissues to use glucose is the basis of a number of diseases, in particular diabetes mellitus. The ability of tissues to utilize glucose is judged by indirect indicators: by the difference in glucose in the blood flowing to and from the organ, by the rate of decrease in glycemia after carbohydrate loading and after administration of insulin, by the amount of glucose in urine with and without carbohydrate load.

 These methods are time-consuming, they are difficult to adapt for mass screening. The authors have not found any methods for directly measuring glucose utilization by tissues suitable for widespread use in medical practice.

 The authors of this method offer experimental studies of academician A.M. Ugolev ("Physiology and pathology of parietal (contact) digestion". L.: Nauka, 1973, pp. 18-36), in which the transition rate of various food substances was studied, including glucose, from solutions of these substances passed through a piece of intestine. These studies made it possible to specify the mechanisms of assimilation of certain food substances by the intestinal wall, however, they are not suitable for implementation in medical practice, since they are too laborious to perform and require extirpation of the studied tissues.

 As a prototype, the authors used a method for determining the utilization of glucose by blood cells placed in an artificially created medium containing a certain amount of glucose. On the rate of utilization, the authors (V. G. Kunitsyn, M. F. Nekrasov, V. I. Fedenkov. “Determining the rate of utilization of glucose in suspension of red blood cells.” Laboratory, 1990, N 9, p. 27-29) judged by the speed loss of glucose from the incubation medium during a certain period of incubation. This study states the possibility of a direct study of the ability of cells to utilize glucose from an artificially created "depot", i.e. from the erythrocyte incubation medium.

 However, for widespread use for diagnostic purposes, the method is not very suitable due to difficulties associated with blood sampling, preparation of a suspension of red blood cells, and their incubation outside the body.

 The authors suggest a method for determining the utilization of glucose by tissues in the gingival fold of the oral cavity. Given that the mechanisms of glucose utilization are universal for all types of tissues (except for the kidneys and small intestine), the results of the method can be extrapolated to the entire body.

 The method proposed by the authors consists of three stages.

1. Preparation of glucose test carriers (TNG). On a piece of filter paper 1.0 • 1.5 cm, glued with waterproof glue to the L-shaped holders and sterilized with dry heat at 120-140 ^o C, 0.02 ml of 8% glucose solution (1.6 mg dry matter) is applied . (V.V. Shkolova, S.V. Shkolova "Test-carrier for determining the intensity of glucose uptake by living tissue". Certificate for utility model N 19469 of September 10, 2001). The quantitative characteristics of the glucose solution applied to TNG are determined experimentally by the authors. After drying in air, the TNG is wrapped in small batches in cellophane with the number and series of the TNG on the handle. TNGs prepared in this way can be stored indefinitely in dry, preferably airtight, pencil cases suitable for transportation.

 2. Exposure of TNG in the upper lip-gingival fold. After 4-fold “cleaning” the folds with the tongue (two clockwise and two counter-clockwise movements), the TNG is inserted into the fold for 10 minutes until it stops. The patient must be warned that it is impossible to allow even a slight displacement of TNG, as this significantly affects the final test result. After exactly 10 minutes, the TNG is removed, the paper part is removed from the holder with tweezers and placed in a test tube containing 1.0 ml of distilled water. The transfer of glucose from the paper part to the solution lasts 10 minutes with constant shaking of the tube, after which the paper part is discarded, and the resulting glucose solution is ready for the next step, i.e., to determine glucose in it. If necessary (for example, during mass testing), TNG after exposure can be dried in air and examined even after a few days.

 3. Determining the glucose content in TNG after transferring it from the paper part to water (immediately after exposure or after some time) can be done by any method of studying the concentration of glucose in aqueous solutions. For example, when quick testing was needed, the authors used the Johnson & Johnson portable one touchscreen basic gyrometer. (Certified by the Ministry of Health and MP RF, registration certificate N 95/160). The degree of glucose utilization by tissues was judged by the difference in its content in TNG after exposure compared with unexposed ones. This difference was expressed as glucose absorption coefficient (GCG), which is the percentage loss of glucose from TNG.

 In order to assess the capabilities of the method, two groups of people were tested.

 1. Nine healthy young people (students) aged 19-25 years old, including 3 men and 6 women (control group).

 2. Eleven women aged 40-70 years, who arrived at different times in the endocrinology department of the city hospital in Tver 6 for the treatment of non-insulin-dependent diabetes mellitus. In the control group, the value of CNG ranged from 12.8 to 65.8. The indicated extreme values of CNG were discarded, and, based on the remaining seven observations, a preliminary “norm” of 45.5 ± 02.3 was obtained. In the group of patients with NIDDM, the value of CNG ranged from 3.8 to 47.4, the average value was 24.2 ± 05.6. From the above data it follows that the degree (speed) of glucose utilization by tissues in both groups varied significantly, however, the general direction of the changes reliably (P <0.05) indicates a weakening of glucose utilization by tissues of patients with NIDDM.

 The authors believe that their proposed method speeds up, reduces the cost and simplifies the procedure for diagnosing diabetes mellitus and carbohydrate metabolism disorders predisposing to the development of this disease, since it eliminates the need for taking and processing blood, urine, stress tests, and allows conducting mass examinations in order to organize preventive work , and also facilitates the operational control of the effectiveness of treatment.

Clinical example N 1
Patient M., 59 years old, turned to the clinic with complaints of pain in the epigastric region and dyspeptic phenomena. An exacerbation of gastric ulcer was diagnosed. A patient has a history of diabetes mellitus. When testing the proposed method, there was a decrease in CNG by 57.6% in comparison with the above-mentioned temporary "norm", which is regarded as an indication of the possibility of registering the proposed method of hereditary characteristics of carbohydrate metabolism, which reduce tissue resistance to damaging factors without the manifestation of diabetic symptoms.

Clinical example N 2
Patient D., 63 years old, was treated at the endocrinology department of the city hospital N 6 in Tver for insulin-independent diabetes mellitus from 09.24.01 to 10.10.01 (medical history N 5784). When hourly determination of glycemia while taking anti-diabetic food additives was conducted parallel testing of the proposed method. The following results were obtained (see table).

 The data obtained are regarded as evidence that an increase in the absorption of glucose by tissues (an increase in CNG) reduces glycemia, and a decrease in absorption leads to an increase in blood glucose.

Claims (1)

A method for determining tissue glucose utilization, including determining the rate of glucose loss, characterized in that 0.02 ml of an 8% glucose solution is applied to the pre-sterilized dry heat at 120-140 ° C, the glucose test carrier (TNG) is placed in the upper the lip-gingival fold after 4 times cleaning it with the tongue, after a 10-minute exposure, the paper part of the TNG is immersed in a test tube with 1.0 ml of distilled water, constantly shaking it, in the aqueous glucose solution formed after 10 minutes, determine its concentration and e attrition glucose compared to the initial amount, determine the absorption coefficient of tissue glucose.

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