A method and a kit for diagnosing type 2 diabetes, metabolic syndrome, sub clinical atherosclerosis, myocardial infarct, stroke or clinical manifestations of diabetes.

A technology for atherosclerosis and type 2 diabetes, which can be used in disease diagnosis, biological testing, material testing, etc., and can solve problems such as unknown causes of increased atherosclerosis

Inactive Publication Date: 2008-04-23
ASTRAZENECA AB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ApoCIII causes hypertriglyceridemia by inhibiting the hydrolysis of triglycerides in very low-density lipoprotein (VLDL), but the cause of its increased LDL atherosclerosis remains unknown

Method used

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  • A method and a kit for diagnosing type 2 diabetes, metabolic syndrome, sub clinical atherosclerosis, myocardial infarct, stroke or clinical manifestations of diabetes.
  • A method and a kit for diagnosing type 2 diabetes, metabolic syndrome, sub clinical atherosclerosis, myocardial infarct, stroke or clinical manifestations of diabetes.
  • A method and a kit for diagnosing type 2 diabetes, metabolic syndrome, sub clinical atherosclerosis, myocardial infarct, stroke or clinical manifestations of diabetes.

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Subject selection

[0054] In the first study, 10 subjects with sdLDL (B phenotype pattern) and subclinical arterial presence in the carotid arteries were compared with 10 age- and sex-matched healthy controls Atherosclerosis without concomitant drug therapy, these 10 controls had no risk factors for metabolic syndrome (NCEP definition plus hyperinsulinemia as a marker of insulin resistance), total cholesterol <6.5mmol / l, no clinical Cardiovascular disease, no subclinical atherosclerosis (no atherosclerotic plaques in the carotid or femoral arteries), and no medication, and clinically healthy (Table 1).

[0055] In the second study, 21 patients with type 2 diabetes randomly selected from the Atherosclerosis and Insulin Resistance Study (Atherosclerosis and Insulin Resistance Study) were compared with 23 age- and sex-matched healthy controls. Insulin Resistance study (AIR), 74 patients in AIR), these 23 healthy controls did not have risk factors for metabolic syndrome, ...

Embodiment 2

[0063] LDL fraction

[0064] LDL densities were isolated from serum samples (1.0-1.35 ml) by ultracentrifugation in a pre-gradient buffer as previously described (Hallberg C, et al., Journal of Lipid Research 1994; 35: 1-9). Subclass, the preformed gradient buffer contains 140mM NaCl, 10mM Na 2 EDTA, Hepes 10mM, pH 7.2, and in different amounts of deuterium oxide (D 2 O) prepared in. Fractions were collected by moving up the gradient. The total protein content of the LDL fraction was determined using the DC protein content assay (Bio-Rad, Hercules, CA, USA) according to the manufacturer's instructions with bovine serum albumin as a standard. ApoB in the LDL fraction was determined by nephelometry using an anti-human apoB antibody (Dakopatts, Denmark). Total cholesterol was determined using a colorimetric method (Roche Diagnostics, RmBH, Manheim, Germany). The densities of the solutions used and the gradient solutions after centrifugation were determined gravimetrically (H...

Embodiment 3

[0067] SELDI analysis of LDL-binding proteins

[0068]In two studies, 50 mM ammonium acetate (pH 6.0) and 50 mM Tris-HCL (pH 9.0) were used on the surface of two types of protein chips (cationic (CM10) protein chip and anionic (Q10) protein chip) LDL fraction 4 (density 1.020-1.040 g / ml) and LDL fraction 5 (density 1.040-1.060 g / ml) from all subjects were analyzed. All samples were processed using a Biomerk laboratory workstation (Beckman-Coulter) modified to utilize the protein chip array bioprocessor (Ciphergen Biosystems). 20 μl of each LDL fraction was mixed with 80 μl of binding buffer, and the mixture was added to the chip surface followed by incubation for 30 minutes. Then the spots were washed three times with 100 μl of binding buffer for 5 min each time to reduce non-specific binding, and finally washed twice with 100 μl of deionized water. Two different types of matrices were used comprising sinapinic acid (SPA) (Aldrich Chem Co, Milw, WI) and alpha-cyano-4-hydroxy...

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Abstract

A method and a kit for diagnosing or prognosing susceptibility to develop type 2 diabetes, the metabolic syndrome, sub clinical atherosclerosis, myocardial infarct, stroke or clinical manifestations of diabetes in a subject.

Description

technical field [0001] The present invention relates to a sensitivity method for diagnosing or predicting clinical manifestations of type 2 diabetes, metabolic syndrome, subclinical atherosclerosis, myocardial infarction, stroke or diabetes in a subject. Background of the invention [0002] Trapping and modifying LDL (low density lipoprotein) particles in the proteoglycan layer of the intima appears to be a key initial step in atherogenesis. Most interactions of LDL with other macromolecules and cells, including those responsible for atherogenesis, are controlled by non-exchangeable segments of apoB100 exposed on the particle surface (Chan L, J Biol Chem 1992; 267:25621-25624; Camejo G, et al, Pathological significance 1998; 139; 205-222). Exchangeable apolipoproteins and proteins appear to mediate some of these interactions when they are adsorbed on the surface of LDL. Effectively defined lipoproteins with a density of 1.019-1.063 are a population of particles of varying ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/92
CPCG01N33/92G01N2333/775G01N2800/042G01N2800/324G01N2800/2871G01N2800/323
Inventor G·卡梅乔P·戴维森J·赫尔塞
Owner ASTRAZENECA AB
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