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Colorimetric determination of fructosamine in hemolytic samples for the postmortem diagnosis of diabetes mellitus.

An analytical method of fructosamine (glycated serum protein) determination in hemolytic samples from cadavers was investigated for the postmortem diagnosis of diabetes mellitus. Fructosamine level is usually measured by the reduction of nitroblue tetrazolium (NBT) using a spectrophotometer. This assay was significantly disturbed by more than 1 g/l hemoglobin, which strongly reduced NBT by the catalytic action of the sulfhydryl and glycated groups. Glycated and total hemoglobin levels were then determined simultaneously to exclude the interferences. Total protein concentration was analyzed to eliminate dilutional effects by hemolysis. With these modifications, corrected fructosamine levels in samples containing not more than 10 g/l hemoglobin could be estimated. The assay of such hemolytic samples from 32 cadavers indicated higher fructosamine values in diabetic group than in non-diabetic subjects and the postmortem degradation of the levels as previously reported.

Adolescent↗

Clinical validation of a second-generation fructosamine assay.

The serum fructosamine assay, used to monitor short-term clinical glycemic control, reportedly has several technical drawbacks. However, technical improvements have resulted in a new second-generation assay of fructosamine. We evaluated this second-generation assay (from Roche Diagnostics) in 529 nondiabetic and diabetic patients and found a highly significant correlation with results of the first-generation assay (r = 0.91, P less than 0.001). Use of the second-generation assay with samples from patients classified on the basis of glycemic control according to their glycohemoglobin (GHb) values, enabled us to discriminate between the nondiabetics, diabetics with "good/moderate" control (i.e., GHb less than 10%), and diabetics with "poor" control (GHb greater than or equal to 10%). We evaluated the validity of the second-generation assay to assess short-term glycemic control in 23 non-insulin-dependent diabetic patients who participated for 10 weeks in an intensive intervention program designed to rapidly normalize the clinical glycemic profile. Results correlated significantly with the one-week average capillary blood glucose concentration (CBG) and with the three-week average CBG in all 23 patients. In addition, the second-generation fructosamine assay results demonstrated a significant decrease at each week of study, as did the average CBG. Results of the first- and second-generation assays correlated significantly at each week of study. GHb correlated significantly with both the second- (r = 0.78, P less than 0.001) and first-generation fructosamine assay results (r = 0.77, P less than 0.001) for the baseline blood samples of the intervention study, but this correlation decreased (to r = 0.35, P = 0.09 and r = 0.34, P = 0.09, respectively) by the conclusion of the study.

Adult↗

Clinical usefulness of serum fructosamine and HbA1 as markers for metabolic control in patients with changing insulin regimens.

The therapy of 10 patients with insulin-dependent diabetes mellitus was changed from insulin twice a day to the use of insulin pens four times a day. The possible changes in glycaemic control were monitored by day curves once a week and analysis of fructosamine and glycated haemoglobin once a fortnight. No significant changes in these parameters were seen. However, a significant decrease in the number of hypoglycaemic periods does point to an improvement of therapy. One patient having elevated values of glucose and glycated haemoglobin but "normal" concentrations of fructosamine prompted us to analyze in detail the influence of protein and albumin on fructosamine. Correction of fructosamine for low protein or albumin increases the value of this test.

Adult↗

[Lipoprotein and apolipoprotein levels in young insulin-dependent diabetic patients. Relations with glycosylated hemoglobin and fructosamine].

The pathogenesis of premature atherosclerosis in diabetic patients has not yet been fully elucidated, but it seems to be related to changes in circulating lipoproteins and to a poor metabolic balance. In this study plasma levels of triglycerides (TG), total cholesterol (TC), HDL- and LDL-cholesterols, apolipoproteins (Apo) A1 and B were measured in 120 young patients aged from 4 to 32 years (mean +/- 1 SD: 17 +/- 6 years) whose diabetes had been present for a mean period of 10 +/- 6 years (range: less than one year to 25 years). The results obtained were analysed in relation to glycosylated haemoglobin (N: 6.8 +/- 0.6 per cent) and plasma fructosamine (N: 1.9 +/- 0.2 mmol/l) levels. The patients were divided into 3 groups according to their HbA 1 level: (group 1: less than 9 per cent, group 2: 9 per cent less than or equal to HbA1 less than 11 per cent; group 3: greater than or equal to 11 per cent. The most significant increases of TG, TC, LDL-C and ApoB levels were observed in group 3, i.e. in patients whose diabetes was the most poorly controlled (HbA 1: 12.9 +/- 1.3 per cent; fructosamine: 4.6 +/- 0.9 mmol/l). These parameters were significantly correlated with HbA1 (p less than 0.01) and even more significantly with fructosamine (p less than 0.001). No significant difference in HDL-C and ApoA1 levels was found in the 3 groups of patients. Thus, TG, TC, LDL-C and ApoB are increased in young diabetics whose HbA1 and fructosamine levels exceed reference values by more than 5 standard deviations.

Adolescent↗

[Routine control of type 2 diabetes. Are glycosylated hemoglobin and fructosamine measurements necessary?].

In 98 type 2-diabetics with stable fasting blood glucose who were treated by tablets and/or diet, fasting blood glucose, glycosylated hemoglobin A1c and serum fructosamine were measured simultaneously. Fasting blood glucose was defined as stable when all values were less than 7.1 mmol/l, or deviated by less than 20% in at least three months, and the treatment had not been changed during the last six months. There is satisfactory correlation between fasting blood glucose and glycosylated hemoglobin A1c (r = 0.64), and serum fructosamine (r = 0.63). It is concluded that glycosylated hemoglobin and fructosamine are superfluous in routine controls of such patients. When fasting blood glucose is unsuitable for practical reasons, fructosamine is a good and cheap alternative. In annual controls glycosylated hemoglobin is the best parameter, and reflects the longest period of metabolic control.

Adult↗

Standardization of serum fructosamine assays.

We have calibrated a secondary serum protein standard by use (as primary standards) of samples of albumin and polylysine glycated with [14C]glucose in vitro, the glycation of which was assessed by radioactivity measurements and by elementary analysis for C and N. Using this standard for calibration in our improved fructosamine assay, one obtains an average fructosamine value of 247 mumol/L for nondiabetic individuals (or, in terms of total serum protein, 3.2 mumol/g)--about a tenth the value we obtained when we used the fructosamine assay of Johnson et al. (Clin Chim Acta 1983;127:87-95), standardized with desoxymorpholinofructose. In contrast, results corresponded well with the value for mean glycation of serum proteins, 3 mumol/g, determined by a furosine/HPLC method. Evidently the proposed procedure, in which a standard sharing the binding characteristics of endogenous glycated proteins is used together with our modified new fructosamine assay, leads to more realistic values for the concentrations of glycated serum proteins.

Carbon↗

[The effect of hyperlipoproteinemia on serum fructosamine].

In 127 patients, we investigated the influence of hyperlipemia on observed fructosamine values. An in vitro influence of the lipids on the fructosamine reading could be excluded for cholesterol, HDL-cholesterol and triglycerides. However, in patients with type I diabetes, both, cholesterol and triglycerides significantly (p less than 0.05) correlated with fructosamine. This may be explained by in vivo effects of hyperglycemia on lipids or lipoproteins. For a given level of hyperglycemia, fructosamine is slightly more sensitive than HbA1c.

Blood Proteins↗

[New formation of fructosamine in patient serum following blood collection].

The stability of fructosamine has been analysed in sera of 24 patients. Fructosamine concentrations increased daily with a constant rate of formation. Synthesis of fructosamine in vitro strongly depended on the incubation temperature and was directly proportional to the glucose concentration of the sample. It is recommended to store sera for the determination of fructosamine at 4 degrees C or - 20 degrees C.

Blood Glucose↗

[Albumin or protein standardized fructosamine-plus, 2 new indices for evaluating diabetic metabolic status].

We have evaluated a new method for the determination of glycated serum proteins (fructosamines), which are elevated in diabetics. In accordance with earlier findings fructosamine depends not only on mean blood glucose but also on albumin- and total protein concentrations. Therefore fructosamine is not useful as an index of diabetic blood glucose control without consideration of an individuals albumin- or protein-concentration. We propose albumin- or protein-standardized fructosamines as new indices of diabetic control.

Blood Glucose↗

[Fructosamine--a dynamic indicator of compensation in diabetes].

The serum fructosamine concentration was evaluated in 22 diabetics on admission to hospital and after intensified treatment with an insulin pump which was indicated on account of decompensation of diabetes. The drop of the mean glucose concentration in blood after 12 days of treatment correlated with the statistically significant drop of the serum fructose concentration (r = 0.69, p less than 0.001). Between the glucose and fructosamine serum concentration no relationship was observed, as long as the mean glucose concentration was calculated for a period shorter than the previous seven days. A highly significant correlation was observed between the rate of fructosamine decline and the mean blood sugar level (r = 0.94, p less than 0.001). From the dynamic changes ensues that when the mean blood sugar level declined by 1 mmol/l, the fructosamine level declined on average by 0.13 mmol/l. The results suggest a more rapid turnover of glycosylated proteins, as compared with data in the literature pertaining to non-glycosylated proteins.

Adult↗

[Comparison of HbA1c, fructosamine and the main metabolic parameters in a non-insulin-dependent diabetic population].

Our objective was the checking of clinical data obtainable from the assay of some parameters in NID diabetic individuals. To this end, we studied 133 patients--57 males and 76 females, average age 74.36 +/- 1.01 years, 72.6% of which were above 65 years of age. The control population was subdivided as follows: 50 subjects, 26 F and 24 M; average age 71.25 +/- 1.32 years, with normal glucidic tolerance as assessed by OGTT. Current glycemia, average glycemia, fructosamine, glycosylated hemoglobin, triglycerides, LDL-cholesterol and apolipoprotein B were obviously much higher than normal in the individuals admitted to the study. A statistically significant correlation was found between average glycemia, glycosylated hemoglobin, LDL-cholesterol and blood triglycerides (p less than 0.05). No correlation was found between current glycemia, fructosamine and glycosylated hemoglobin. Similarly, serum fructosamine was unrelated to the parameters studied. In our study, fructosamine, glycosylated hemoglobin and current glycemia offered unrelatable data. Hence, in our opinion it is necessary to assay these three parameters contemporaneously for a reliable assessment of metabolic compensation.

Aged↗

[Value of plasma fructosamine in non-diabetic and diabetic chronic renal failure].

Plasma fructosamine concentrations were measured in non-diabetic and diabetic patients with chronic renal failure divided into two three groups: patients without dialysis, under haemodialysis and under continuous ambulatory peritoneal dialysis. In non-diabetic patients plasma fructosamine values were consistently higher than in a control population (2.26 +/- 0.26 mmol/l), being 2.38 +/- 0.35 mmol/l in patients without dialysis, 2.57 +/- 0.33 mmol/l in patients under peritoneal dialysis and 2.67 +/- 0.31 mmol/l in patients under haemodialysis. In diabetic patients, plasma fructosamine values were increased, being equal to, or higher than 3 mmol/l; these values were almost identical with those obtained in populations of diabetics without renal pathology. Considering that Hb Alc values are difficult to interpret in chronic renal failure owing to anaemia and to the analytical problems raised by haemoglobin carbamylation, the fructosamine test may well be a reliable marker for the monitoring of diabetes in patients with chronically impaired renal function.

Diabetes Mellitus, Type 1↗

[Assay of fructosamine. Value and limits in diabetology].

Fructosamine test using a Nitroblue Tetrazolium (NBT) method offers many advantages: quickness, reproducibility, easy automation and unexpansiveness, but a standardization of the different methods is needed. The results can be expressed in absolute value of equivalent DMF per liter, except in pregnancy where mumol per g of protein is used. The interpretation of the results can be difficult in case of quantitative and/or qualitative proteins abnormalities: icterus and severe chronic renal insufficiency. Fructosamine is significantly higher in diabetic patients. It gives a good correlation with glycated haemoglobin but the provided information is different, concerning a shorter period of 2 to 3 weeks and perhaps more sensitive to recent glycaemic variations. Fructosamine test does not seem to be a good screening test for diabetes and impaired glucose tolerance. The test indications of the assay could be: situations where the dosage of glycated haemoglobin is not interpretable, diabetic pregnancy follow-up and short term evaluation of a therapeutic change on glycaemic control. However, the individual significance of fructosamine concentration remains to be assessed and seems to be less accurate than glycated albumin.

Biological Assay↗

[Clinical significance of the determination of serum fructosamine and albumin in children with diabetes mellitus].

The paper is devoted to a study of the role of serum glycoprotein fructosamine and serum albumin in the pathogenesis of a severe course of insulin dependent diabetes mellitus (IDDM) in children. Fructosamine was determined in 43 pediatric patients with IDDM by direct spectrophotometry using Hoffman-La-Roche kits; albumin, C-peptide and malonic aldehyde were also determined. Disorder of the mechanism of regulation of homeostasis by albumin was shown to play an important role in the pathogenesis of a severe course of IDDM in children. It could be caused by its enhanced glycosylation and a decrease in liver synthesis in some cases as a result of considerable reduction of insulin secretion. A prognostically unfavorable sign was a raised ratio of fructosamine to albumin and enhanced lipid peroxidation against a background of low insulin secretion. The determination of serum levels of fructosamine and albumin can be a valuable diagnostic criterion in examination of children with diabetes mellitus.

Biomarkers↗

[Evaluation of the assay of serum fructosamine in the surveillance of diabetes in children and adolescents].

Serum fructosamine and glycosylated haemoglobin were measured 4 times over 2 month period (1st, 15th, 30th and 60th days) in 40 young diabetics and in 26 controls. A positive correlation was found between these two parameters at the 15th, 30th and 60th days (0.44 less than r less than 0.61, P less than 0.01). There was a milder correlation between glycosylated haemoglobin and fasting glycemia (r = 0.43), between serum fructosamine and mean glycemia of the last 4 weeks (r = 0.38) and between serum fructosamine and 24 h glycosuria (r = 0.39) (P less than 0.05). Serum fructosamine, which reflects the mean glycemia of the last 3 weeks appears as a useful parameter in diabetic control, in association with glycosylated haemoglobin which reflects the mean glycemia of the last 2 months.

Adolescent↗

Effects of various serum proteins on quantification of fructosamine.

We determined fructosamine concentrations with the CentrifiChem 600 centrifugal analyzer and the Hitachi 737 discrete analyzer. Reference intervals agreed with the most recently published results, and values in fasting patients were significantly correlated with glycated hemoglobin, plasma glucose, albumin, beta- and gamma-globulins, IgG, IgA, IgM, and total protein. Partial correlation analysis showed that only fructosamine and IgA were dependently related. In a group of nondiabetic patients with pathological values for IgA concentrations, 79.4% had pathological values for fructosamine. These results throw doubt on the clinical value of fructosamine determinations if serum IgA is not taken into account.

Adult↗

[Fructosamine, a new marker of blood glucose control in the diabetic].

Serum fructosamine assay is a new, simple and fast method used to evaluate serum glycosylprotein concentrations. We performed this assay in 96 healthy controls, 95 patients with diabetes (insulin-dependent in 71, non insulin-dependent in 24) and 23 non diabetic pregnant women. Serum fructosamine concentrations were increased and correlated with percents of glycosyl haemoglobin in all diabetics, irrespective of the type of diabetes. However, the low correlation coefficient and the fact that some individual values plotted were distant from the regression slope suggested that fructosamine does not have the same biological significance as glycosyl haemoglobin: the formation and degradation kinetics of these two substances are known to be very different. Fructosamine values were tightly close together in controls and in pregnant women, the latter showing lower values.

Adolescent↗

Is serum fructosamine assay specific for determination of glycated serum protein?

We compared the fructosamine activity in sera from healthy and diabetic subjects with the degree of protein glycation detected by a liquid-chromatographic method. The latter technique measures furosine as a specific product after hydrolysis of epsilon-amino-fructose-lysine. Our results indicate that the fructosamine assay measures the extent of glycation of purified human serum albumin correctly. On the other hand, we found no correlation between the two methods for sera from healthy subjects, although for diabetics' sera the values obtained with both methods were related. However, only about half of the reducing activity (fructosamine) was due to specific nonenzymatic glycation of proteins in healthy subjects and well-controlled diabetics. The remaining unspecific activity varied from serum to serum. It was not reducible with NaBH4 and was independent of the glycation of albumin, which normally accounts for about 80% of glycated serum proteins. The fructosamine assay is therefore of limited specificity for the exact measurement of glycated proteins in serum.

Blood Proteins↗