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Influence of triglycerides and urate on methods for determination of fructosamine.

The original fructosamine assay, based on reduction of NBT, has been modified and marketed as a kit. The development includes addition of detergents, uricase, change of the buffer and NBT concentrations, and use of a new calibrator. We report the effects of these changes on the measured values of fructosamine in lipemic and uricemic sera. A secondary calibrator produced from pooled serum and stabilized by removal of glucose was used to calibrate the original fructosamine assay and enhance transferability of results. There was no correlation between the difference between the results obtained by the two procedures and the concentration of urate or triglycerides. The development of colour was faster with the original method. It is suggested that measurements of fructosamine by either method are practically equivalent. However, the use of polylysine as a calibrator is advantageous. It is suggested that the generic term S-Glycated proteins is used for the measurand rather than S-Fructosamine which in effect has become a commercial term.

Fructosamine↗

Comparison of reliability of plasma fructosamine and glycosylated hemoglobin assays for assessing glycemic control in diabetic patients on hemodialysis.

To search for a reliable marker of medium-term integrated blood glucose level in diabetics on maintenance hemodialysis (HD), plasma fructosamine and glycosylated hemoglobin (Hb) levels were determined every week and blood glucose levels were determined four times every day over 3 weeks. The mean values of fructosamine (mol/L per 40 g of albumin) and of glycosylated Hb of other the study period correlated (r = .746, P less than .001) for combined materials of diabetic and nondiabetic subjects. However, plasma fructosamine values at the end of the study period did not correlate with the overall mean blood glucose values during the preceding 8 to 21 days (r = .372, NS). In contrast, glycosylated Hb values correlated closely with the same mean blood glucose values (r = .703, P less than .001). Fructosamine values significantly decreased during a HD, irrespective of the increases in albumin and total protein. In conclusion, glycosylated Hb was a reliable marker of long-term integrated blood glucose even in diabetics on HD. However, fructosamine was not a reliable marker of medium-term integrated blood glucose in these patients.

Blood Glucose↗

Elimination of superoxide dismutase interference in fructosamine assay.

OBJECTIVE: Superoxide dismutase (SOD) (EC 1.15.1.1) is reported to decrease the reduction of nitroblue tetrazolium (NBT) in the fructosamine assay. The study was undertaken to find a method to eliminate this interference. DESIGN AND METHODS: We studied the NBT reduction in the presence and absence of added bovine erythrocyte SOD during fructosamine assay. Formation of reduced NBT decreased with the increasing concentration of SOD. Various inhibitors of SOD were experimented with for effectively eliminating this interference. RESULTS: Cyanide eliminates the interference due to SOD, but is unsuitable because in it's presence glucose becomes reducing under the conditions of fructosamine assay. SOD inhibitors such as EDTA and Azide did not eliminate the effect of SOD. Guanidine. HCl gives opalescence in the reaction mixture. Addition of 2M HCl to the serum and incubation at 37 degrees C for 10 min eliminated the effect of added SOD (70 kU/L). The correlation between deltaA10-20 min of serum treated with HCI in presence and absence of added SOD is y = 0.9011x + 0.01055 with r = 0.9789 and S.E. (y) 0.007686. CONCLUSION: SOD does not interfere in globin bound fructosamine assay as acid-acetone treatment in preparation of heme free globin inhibits SOD. Pretreatment of serum with HCl can satisfactorily eliminate the interference due to SOD in fructosamine assay. The acid treatment could be used to inhibit SOD in various other reactions that are followed with NBT reduction.

Animals↗

Fructosamine in diabetic pregnancy.

Fructosamine, an indicator of glycosylated serum protein, was measured in 79 non-diabetic pregnant women and 20 women with gestational diabetes. The test provided a clear discrimination between groups; it detected 17 (85%) of the women with gestational diabetes and gave only 4 (5%) false-positive results. 19 women with established diabetes before pregnancy had very high levels. Maternal fructosamine at 29 weeks' gestation correlated significantly with both fasting blood glucose levels at the time and birthweight ratio. Levels of fructosamine in cord blood were significantly higher in gestational diabetic than in normal pregnancies, suggesting a possible additional role for fructosamine in retrospectively detecting the hyperglycaemic fetus. Fructosamine estimation is fully automated and may provide a simple, inexpensive means to screen for diabetes in pregnancy.

Adult↗

Serum fructosamine concentrations in dogs with hypothyroidism.

Serum fructosamine concentrations were measured in 11 untreated hypothyroid dogs with normal serum glucose and serum protein concentrations. The fructosamine level ranged between 276 and 441 micromol/L (median 376 micromol/L; reference range 207-340 micromol/L). Nine of the 11 dogs had fructosamine levels above the reference range. The fructosamine levels decreased significantly during treatment with levothyroxine. It is suggested that serum fructosamine concentrations may be high in hypothyroid dogs because of decelerated protein turnover, independent of the blood glucose concentration.

Aging↗

Serum fructosamine concentrations in hyperthyroid cats.

Serum fructosamine concentrations were measured in 35 healthy cats and in 30 hyperthyroid cats before and 30 days after curative radioiodine ((131)I) treatment. Hyperthyroid cats were divided into those with 30 day post-treatment total thyroxine (T4) concentrations within (EuT4) or below (HypoT4) the reference range. The median (semi-interquartile range, SIR) fructosamine concentration was significantly lower in hyperthyroid compared with healthy cats (295. 0 (18.5) micromol l(-1)) both before (254.0 (27.6) micromol l(-1)) and after (268.5 (28.0) micromol l(-1)) treatment (P < 0.001 in each case). (131)I therapy was associated with increases in serum fructosamine (mean increase 20.4 micromol l(-1), P = 0.039) and total protein (6.3 g l(-1), P < 0.002) in the HypoT4 group and in globulin concentration in both EuT4 (5.9 g l(-), P < 0.002) and HypoT4 (5.2 g l(-1), P = 0.023) groups. There were no direct relationships between the observed elevations in fructosamine concentration and those in total protein or globulin concentrations suggesting that the effect may be due to reduced rates of protein turnover. Reduced values may need to be considered when interpreting serum fructosamine concentrations for monitoring the degree of glycaemic control in diabetic cats with concurrent hyperthyroidism.

Animals↗

Serum fructosamine (alkaline-reducing activity) for evaluation of glycemic control in diabetic patients.

The values of serum fructosamine were significantly higher in 75 diabetic patients than those of metabolically healthy controls. Significant correlation could be observed between the glycated hemoglobin and serum fructosamine values in diabetics. Furthermore, significant correlations were established between the serum fructosamine values and the mean blood glucose as well as the mean 24 hours urinary glucose values measured 10-14 days previously in diabetics. Glycated hemoglobin values should be measured in diabetics for long-term assessment of glycemic control while serum fructosamine values could be useful in clinical practice for assessment of overall glycemic control of the previous two weeks in diabetic patients. Not only long-term glycemic control but medium-term alterations in blood glucose concentrations could be evaluated by simultaneous measurements of glycated hemoglobin and serum fructosamine values in diabetics.

Adult↗

Effect of thiols on fructosamine assay.

Fresh human serum, gGAPDH and beta-mercaptoethanol were used to examine the effect of thiols on fructosamine assay. The kinetics of the reaction with both fresh human serum and gGAPDH displayed biphasic behaviour (fast and slow). When the thiols were modified with IAA, the kinetics only demonstrated the slow phase. Since the absorbance increase in the interval from 9 to 10 min was used in the fructosamine assay of glycated human serum we studied the effect of thiols on that measurement. In the case of gGAPDH, the value was approximately one-half of the original after thiol modification, suggesting thiol interference. Nevertheless, gGAPDH may contain a fructosamine structure. beta-Mercaptoethanol itself gave a strong positive result in the fructosamine assay. Hence, thiol groups on glycated proteins should be modified before doing a fructosamine assay because of their substantial interference.

Blood↗

Glycated haemoglobin and fructosamine in non-diabetic subjects with chronic renal failure.

Measurements of glycated haemoglobin by electroendosmotic and chromatographic methods, fructosamine, and fructosamine:albumin ratio were made in 91 non-diabetic subjects with chronic renal failure managed conservatively (n = 25), by continuous ambulatory peritoneal dialysis (n = 22), by haemodialysis (n = 22), or by renal transplantation (n = 22). Results were compared with those in a control group of 43 non-diabetic subjects with normal renal function. Mean glycated haemoglobin measured by electroendosmosis was significantly greater in all groups with chronic renal failure except the transplant group. Mean glycated haemoglobin measured by affinity chromatography was not significantly different from controls in any of the groups with chronic renal failure. No difference in mean fructosamine concentration was detected in the transplant or conservatively managed groups compared to controls, but values were significantly lower in the CAPD group, and greater in the haemodialysis group predialysis. Post-haemodialysis samples showed a significant reduction in mean fructosamine concentration when compared with prehaemodialysis samples. Fructosamine:albumin ratios were elevated in all groups of patients with renal failure, with the exception of the transplant group. Of the four indices of glycaemic control considered in this study, only glycated haemoglobin measured by affinity chromatography appears to be unaffected by chronic renal failure.

Adolescent↗

Mechanism of the degradation of non-enzymatically glycated proteins under physiological conditions. Studies with the model fructosamine, N epsilon-(1-deoxy-D-fructos-1-yl)hippuryl-lysine.

The degradation of fructosamines, formed from the non-enzymic glycation of proteins under physiological conditions, to advanced glycation end products was investigated by studying the model peptide fructosamine N epsilon-(1-deoxy-D-fructos-1-yl)hippuryl-lysine (DHL). At pH 7.4 and 37 degrees C in aerobic phosphate buffer, DHL degraded to form N epsilon-carboxymethyl-hippuryl-lysine, and hippuryl-lysine over a 29-day incubation period. The expected N epsilon-(3-lactato)hippuryl-lysine and 'hippuryl-lysylpyrraline' derivatives were not found. Superoxide radicals and hydrogen peroxide were formed during the degradation of DHL but were also both consumed during the degradation reaction. Reversal of the Amadori rearrangement was not a major fate of the fructosamine. The formation of N epsilon-carboxymethyl-hippuryl-lysine was decreased by desferrioxamine, catalase, superoxide dismutase, catalase with superoxide dismutase, anaerobic conditions and aminoguanidine. The formation of hippuryl-lysine was decreased by desferrioxamine, catalase and catalase with superoxide dismutase, but was increased by the addition of aminoguanidine. N epsilon-Carboxymethyl-serine and unmodified lysine residues are major peptide-based end products in the degradation of lysyl-fructosamine under physiological conditions. Oxygen, redox-active metal ions, catalase, superoxide dismutase and the pharmacological agent aminoguanidine are expected to be influential on the rate and fate of fructosamine degradation.

Anaerobiosis↗

Serum fructosamine as a screening test for diabetes mellitus in non-healthy middle-aged to older dogs.

Diabetes mellitus is a common endocrine disorder among non-healthy middle-aged to older dogs. Previous studies have shown that serum fructosamine can be used to detect the chronic unregulated hyperglycaemia in Diabetes mellitus. The present study was therefore conducted to assess the value of serum fructosamine as a screening test for Diabetes mellitus among non-healthy middle-aged to older dogs in terms of sensitivity, specificity, positive and negative predictive values. A total of 99 non-healthy dogs (16 dogs with Diabetes mellitus and 83 dogs with other diseases) with an age ranging from 6 to 15 years were included in the study. Using the upper reference limit for serum fructosamine (343.8 mumol/l), sensitivity and specificity were 0.88 and 0.99, respectively and assuming the prevalence of Diabetes mellitus to be 0.05, the positive and negative predictive values were 0.82 and 0.99, respectively. Apparently, serum fructosamine could also be used to distinguish hyperglycaemia non-diabetic dogs from hyperglycaemic diabetic dogs. Thus, the present study showed that serum fructosamine is a reliable screening test for Diabetes mellitus among non-healthy middle-aged to older dogs.

Aging↗

Clinical utility of assays of glycosylated haemoglobin and serum fructosamine compared: use of data on biological variation.

Assessment of the relative clinical usefulness of glycosylated haemoglobin (HbA1) and serum fructosamine is complicated by their markedly different half-lives. They have therefore been compared using a number of indices derived from data on biological variation, as applied in a study of blood glucose control in newly diagnosed diabetic patients. The within-subject (CVI) and between-subject (CVG) variation of fructosamine and HbA1 were assessed in 8 stable diabetic patients. A slightly smaller relative change, or critical difference, is required between serial HbA1 results than between fructosamine results before a significant change can be said to have occurred. The heterogeneity of within-subject variance is also less for HbA1, rendering the critical difference more generally applicable. HbA1 may therefore be more appropriate for long-term monitoring. Monitoring of blood glucose control of a group of 26 newly diagnosed diabetic patients before the commencement of therapy and 1, 2, and 3 months later by serum fructosamine or HbA1 provided very similar information. Fructosamine responds more rapidly with changes of blood glucose control. However, whether this confers any clinical advantage will depend upon the frequency of testing.

Aged↗

Serum fructosamine as a screening test for diabetes in the elderly: a pilot study.

OBJECTIVE: To determine the value of serum glycated protein, measured as serum fructosamine, as a screening test for diabetes in the elderly. DESIGN: Cross-sectional pilot study. SETTING: Ambulatory research clinic in university setting. PATIENTS: One hundred fifty-seven consecutive community-dwelling participants in the Cardiovascular Health Study, average age 71.8 + 5 (mean +/- SD, range 65-88 years). MEASUREMENTS: Serum fructosamine levels (first and second generation assay) were obtained. All subjects who did not have a diagnosis of diabetes were given a 75-g glucose tolerance test (GTT). RESULTS: Twenty-six subjects (17%) (10 previously diagnosed, 16 undiagnosed and asymptomatic) had diabetes mellitus, and 38 subjects (24%) had impaired glucose tolerance by history or by the GTT (WHO criteria). Only the 16 asymptomatic diabetics were included in the analysis for the pilot study. There was a significant difference in the fasting fructosamine level between non-diabetics and asymptomatic diabetics for the first generation (2.06 +/- .21 vs 2.53 +/- .49 mMol/L, P < 0.0015) and second generation assay (221 +/- 27 vs 269 +/- 48 mMol/L, P < 0.0012). Receiver operator curves were constructed to evaluate the test characteristics of serum fructosamine. Using a point of > or = 2.3 mMol/L for the first-generation assay, the sensitivity to detect asymptomatic diabetes was 75%, specificity 83%, and positive predictive value 35%. To detect both diabetes and impaired glucose tolerance using a cutpoint of > or = 2.3 mMol/L, the sensitivity was 24%, specificity 95%, and positive predictive value 68%. Employing a cut point of 250 muMol/L for the second generation assay, the sensitivity to detect diabetes was 81%, specificity 87%, and positive predictive value 43%. However, to detect diabetes and glucose intolerance using the second generation assay, the sensitivity was 39% and specificity was 86%. CONCLUSION: This study demonstrated that a single measurement of either first or second generation fructosamine showed promise as a screening test for diabetes, but not impaired glucose tolerance, in older people.

Aged↗

Clinical usefulness of estimation of serum fructosamine concentration as a screening test for diabetes mellitus.

Fructosamine, a putative measure of serum glycosylated proteins, was measured in 74 subjects referred for oral glucose tolerance tests. A normal range (mean (2 SD] of 1.6 (0.4) mmol/l (40(10) mg/100 ml) derive from results obtained in 83 healthy non-diabetic volunteers permitted the detection of 15 out of 17 (88%) subjects with proved diabetes and yielded only five (9%) false positive diagnoses. Fructosamine concentrations correlated significantly (p less than 0.001) with fasting plasma glucose concentrations (r = 0.76) and glycosylated haemoglobin concentrations (r = 0.70). A longitudinal study suggested that fructosamine concentration was an index of intermediate term (one to three weeks) blood glucose control. Fructosamine concentration was not related to uraemia and did not depend on albumin or total protein concentrations, provided that serum albumin concentrations remained above 30 g/l. Estimation of fructosamine concentrations is a fully automated procedure and may provide a simple means of screening for diabetes mellitus.

Adolescent↗

Serum fructosamine concentration as measure of blood glucose control in type I (insulin dependent) diabetes mellitus.

Serum fructosamine activity was studied in 42 patients with type I (insulin dependent) diabetes mellitus and 30 non-diabetic volunteers as an index of blood glucose control. There was a significant correlation both between fructosamine and glycosylated haemoglobin values (r = 0.82) and between fructosamine and the fasting C peptide concentration (r = -0.81). Test results in 14 of the diabetics reflected the mean plasma glucose concentration calculated from 25 serial estimations in a single 24 hour period (r = 0.75; p less than 0.01) but not the mean amplitude of glycaemic excursion (r = 0.23; p greater than 0.05). Fructosamine concentrations measured in these multiple blood specimens did not change significantly throughout the day (mean coefficient of variation 4.1%) despite wide variability of the respective plasma glucose concentrations (mean coefficient of variation 36.2%). It is concluded that a single random serum sample analysed for fructosamine concentration provides a simple and reliable assessment of glucose homoeostasis in patients with type I diabetes mellitus.

Adolescent↗

Does the measurement of serum fructosamine accurately reflect levels of glycated albumin in insulin-dependent diabetes?

We have measured serum glycated albumin (GSA) by affinity chromatography and immunoturbidimetry, and serum fructosamine using a Cobas FARA analyser in blood samples from 37 type I diabetics and 21 healthy controls. Random blood glucose and glycated haemoglobin levels were also measured. Glycated haemoglobin (HbA1) correlated with glycated albumin and fructosamine in the diabetic group. A less clear relationship was found between glycated albumin and fructosamine. HbA1, GSA and fructosamine correlated poorly with random blood glucose levels. These data indicate that serum fructosamine levels do not accurately reflect those of glycated albumin, as has recently been suggested, in type I insulin-dependent diabetics where glycaemic control fluctuates more than in type II diabetics. It is postulated that the two methods reflect varying glycaemic levels to a different degree, thereby accounting for the disparity.

Adolescent↗

Influence of serum protein levels on serum fructosamine levels.

The serum fructosamine level is influenced by the serum protein level. Therefore, we investigated the relationship between serum fructosamine levels and serum total protein and albumin levels measured simultaneously in non-diabetic and diabetic patients. Significant positive correlations were found between them. A correction formula was constructed to express the serum fructosamine level when the serum total protein level was 7 g/dl or when the serum albumin level was 4 g/dl: corrected fructosamine (mmol/l) = measured fructosamine (mmol/l) + A (7 (g/dl) or 4 (g/dl)--serum total protein (g/dl) or serum albumin (g/dl] where A is 0.28 for serum total protein and 0.30 for serum albumin in non-diabetic subjects; 0.29 for serum total protein and 0.31 for serum albumin in non-diabetic and diabetic subjects combined.

Blood Glucose↗

Relative clinical usefulness of glycosylated serum albumin and fructosamine during short-term changes in glycemic control in IDDM.

Serial changes in glycosylated blood proteins and direct measures of glycemia were studied in 100 subjects with insulin-dependent diabetes mellitus (IDDM) over a 6-wk period while attempts were made to improve glycemic control. All measures of glycemic control improved significantly (P less than .001). Mean +/- SE glycosylated hemoglobin (HbA1) fell from 9.1 +/- 0.2 to 8.0 +/- 0.1%, glycosylated serum albumin (GSA) from 9.8 +/- 0.4 to 7.3 +/- 0.3%, and fructosamine from 3.92 +/- 0.08 to 3.42 +/- 0.07 mM. Fasting blood glucose levels fell from 11.1 +/- 0.6 to 8.1 +/- 0.7 mM mean blood glucose levels from 12.5 +/- 0.3 to 8.8 +/- 0.03 mM, and the M value from 118 +/- 7 to 40 +/- 3 U. Mean percentage changes in direct measures of glycemia (32-66%) and GSA (29%) were greater than for fructosamine (11%) or HbA, (12%) levels (P less than .001). Furthermore, the correlation between the change in GSA and changes in direct measures of glycemia over the initial 2-wk period was significantly different from the corresponding correlations between direct measures of glycemia and fructosamine over this period (P less than .05-.01). Changes in GSA also correlated more closely than HbA1 or fructosamine did with direct measures of glycemia after 4 and 6 wk. The Spearman rank-correlation coefficient (rs) of absolute changes in GSA, fructosamine, and HbA1 after 2-6 wk ranged from 0.27 to 0.57, confirming that the three measures responded differently to changing glycemic control.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗