Study of the effect of total serum protein and albumin concentrations on canine fructosamine concentration.

The relationship among serum fructosamine concentration and total serum protein and albumin concentrations were evaluated in healthy and sick dogs (diabetics and dogs with insulinoma were not included). Fructosamine was determined using a commercial colorimetric nitroblue tetrazolium method applied to the Technicon RA-500 (Bayer). Serum fructosamine concentration was not correlated to total protein in normoproteinemic (r = 0.03) and hyperproteinemic dogs (r = 0.29), but there was a high correlation (r = 0.73) in hypoproteinemic dogs. Similar comparison between serum fructosamine and albumin concentrations showed middle correlation (r = 0.49) in normoalbuminemic dogs and high degree of correlation (r = 0.67) in hypoalbuminemic dogs. These results showed the importance of recognizing serum glucose concentration as well as total serum protein and albumin concentrations in the assay of canine serum fructosamine concentration.     

Insulinoma in a normoglycaemic dog with low serum fructosamine

An insulinoma was diagnosed in an eight-year-old, male golden retriever. Four fasted serum glucose concentrations were within the reference values over a seven-week period. Two serum fructosamine values over the same seven-week period were both low. An exploratory laparotomy and histopathology of an excised pancreatic nodule confirmed the diagnosis of insulinoma. This report demonstrates that a dog with a histologically confirmed insulinoma can have normal fasting serum glucose concentrations on multiple assays. It also demonstrates that fructosamine assays may be helpful in the diagnostic evaluation of normoglycaemic dogs with clinical signs suggestive of insulinoma.     

Relation of fructosamine to serum protein, albumin, and glucose concentrations in healthy and diabetic dogs.

The relation of the glycated serum protein, fructosamine, to serum protein, albumin, and glucose concentrations was examined in healthy dogs, dogs with hypo- or hyperproteinemia, and diabetic dogs. Fructosamine was determined by use of an adaptation of an automated kit method. The reference range for fructosamine in a composite group of control dogs was found to be 1.7 to 3.38 mmol/L (mean +/- SD, 2.54 +/- 0.42 mmol/L). Fructosamine was not correlated to serum total protein, but was highly correlated to albumin in dogs with hypoalbuminemia. To normalize the data with respect to albumin, it is suggested that the lower limit of the reference range for albumin concentration (2.5 g/dl) be used for adjustment of fructosamine concentration and only in hypoalbuminemic dogs. In 6 hyperglycemic diabetic dogs, fructosamine concentration was well above the reference range. It is concluded that although fructosamine may be a potentially useful guide to assess the average blood glucose concentration over the preceding few days in dogs, further study is required to establish its value as a guide to glucose control in diabetic dogs.     

An evaluation of serum fructosamine as a marker of the duration of hypoproteinaemic conditions in dogs

Serum samples were collected from 153 normoglycaemic, hypoproteinaemic dogs of known case histories, and assayed for fructosamine, glucose, total protein and albumin concentrations. This study was conducted to evaluate the relationship between serum fructosamine and total serum proteins, or more specifically serum albumin. Serum fructosamine was positively correlated with both total serum protein (r=0.47, p>0.00001) and serum albumin (r=0.77, p>0.00001). Mean serum albumin concentrations were significantly different when the data were grouped as dogs with normal versus subnormal serum fructosamine concentrations. The data indicate the value of the serum fructosamine assay in estimating the duration of hypoalbuminaemia. Concurrent hypoalbuminaemia and normal serum fructosamine indicate hypoalbuminaemia of less than one week. Concurrent hypoalbuminaemia and hypofructosaminaemia indicate persistent hypoalbuminaemia of more than one week, and concurrent normal albumin and hypofructosaminaemia indicate recovery from a condition including hypoalbuminaemia or hypoglycaemia.     

Serum fructosamine measurement: a new diagnostic approach to renal glucosuria in dogs

Measurement of serum fructosamine, 1-amino-1-deoxyfructose, is commonly used in diagnosing and monitoring hyperglycaemic disorders, such as diabetes mellitus in dogs. Serum fructosamine indicates long-term serum glucose concentrations and replaces serial serum glucose measurements. This study investigates the clinical usefulness of serum fructosamine in differentiating conditions other than diabetes mellitus characterised by glucosuria. Four dogs presented with glucosuria all had serum fructosamine concentrations within or close to the reference range (313 micromol 1(-1), 291 micromol 1(-1), 348 micromol 1(-1), 262 micromol 1(-1) reference range: 250 to 320 micromol 1(-1) indicating that a single serum fructosamine measurement is a simple and efficient way of verifying concurrent persistent normoglycaemia. Therefore, serum fructosamine is a useful parameter not only in diabetic patients, bu also in differentiating conditions in dogs characterised by glucosuria without hyperglycaemia, such as primary renal glucosuria and the Fanconi syndrome. To distinguish between primary renal glucosuria and the Fanconi syndrome, measurement of the amino acid concentration in urine was performed.     

Evaluation of two point-of-care analysers for measurement of fructosamine or haemoglobin A1c in dogs

Measurement of glycosylated proteins such as fructosamine and haemoglobin A1c (HbA1c) can be used to assess glycaemic control in canine diabetic patients. Two point-of-care analysers, designed for human diabetics, were evaluated for use in dogs. Blood samples were collected from 50 normoglycaemic dogs, 100 diabetic patients and five dogs with insulinoma and tested using the In Charge fructosamine meter and the Haemaquant/Glycosal HbA1c meter. Readings were obtained in all cases except for 21 of 50 diabetics, which were above the upper limit of the In Charge meter. Diabetic dogs had higher fructosamine and HbA1c concentrations compared to controls. However, there was poor agreement between the In Charge meter readings and serum fructosamine concentrations, suggesting that there are problems associated with the use of this device in dogs. HbA1c concentrations showed a high degree of correlation with glycosylated haemoglobin measured at an external laboratory, suggesting that the Haemaquant/Glycosal meter warrants further evaluation for veterinary use.     

Various protein and albumin corrections of the serum fructosamine concentration in the diagnosis of canine diabetes mellitus

Fructosamines are formed when glucose reacts non-enzymatically with amino groups on proteins, and previous studies have indicated that the serum fructosamine concentration could be of importance in the diagnosis of canine diabetes mellitus. Owing to the connection between the protein/albumin concentration and serum fructosamine concentration, it has been suggested that the serum fructosamine concentration should be corrected for the protein/albumin concentration. Thus, the purpose of the present study was to evaluate the uncorrected serum fructosamine concentration and various protein and albumin corrections of the serum fructosamine concentration in the separation of dogs with diabetes mellitus from dogs with other diseases that presented with clinical signs suggestive of diabetes mellitus. The evaluation was assisted by relative operating characteristic curves (ROC curves), which may be used to compare various diagnostic tests under equivalent conditions (equal true positive ratios or false positive ratios) and over the entire range of cutoff values. A total of 58 dogs (15 dogs with diabetes mellitus and 43 dogs with other diseases) were included in the study. Serum fructosamine concentration, serum total protein concentration and serum albumin concentration were measured in each dog, and various corrections of the serum fructosamine concentration for protein or albumin concentration were made. Comparing the ROC curves of the uncorrected and each corrected serum fructosamine concentration indicated that there was no decisive difference between the uncorrected and the corrected serum fructosamine concentrations in discriminating between dogs with and without diabetes mellitus. Hence, correcting the serum fructosamine concentration as a routine procedure cannot be advocated from the results of the study. Moreover, the sensitivity and specificity of the uncorrected serum fructosamine concentration were very high, 0.93 and 0.95, respectively, further evidence of the value of the uncorrected serum fructosamine concentration in the diagnosis of canine diabetes mellitus.Abbreviations SFC serum fructosamine concentration - SFC-P serum fructosamine concentration corrected for the actual serum total protein concentration - SFC-A serum fructosamine concentration corrected for the actual serum albumin concentration - SFC-Po serum fructosamine concentration corrected for the actual serum total protein concentration, only when the serum total protein concentration is outside the reference interval - SFC-Ao serum fructosamine concentration corrected for the actual serum albumin concentration, only when the serum albumin concentration is outside the reference interval - SFC-K serum fructosamine concentration corrected according to Kawamotoet al. (1992)     

Fructosamine and glycated hemoglobin in the assessment of glycaemic control in dogs

Fructosamine and glycated hemoglobin (HbA1c) concentrations were measured simultaneously in 222 dogs (96 healthy and 126 sick dogs). The dogs were divided into 3 groups according to the glucose concentration: hypo, hyper and euglycaemic dogs. Serum fructosamine concentrations were measured by the reduction test with nitroblue tetrazolium. A turbidimetric inhibition immunoassay and specific polyclonal antibodies were used to evaluate glycated hemoglobin concentrations. A significant correlation was found between glucose concentration and either fructosamine (r = 0.63, p < 0.0001) or glycated hemoglobin (r = 0.82, p < 0.0001). The correlation was higher in hyperglycaemic dogs for fructosamine (r = 0.80, p < 0.0001) and in hypoglycaemic dogs for glycated hemoglobin (r = 0.91, p < 0.005). We found a significant correlation between serum fructosamine and glycated hemoglobin (r = 0.65, p < 0.0001 ) when all the dogs were studied. A significant correlation was observed between serum fructosamine and glycated hemoglobin only in hyperglycaemic dogs (r = 0.82, p < 0.0003). Thus, fructosamine and HbA1c may be considered for use in screening tests for diabetes mellitus in dogs and clinical tests for monitoring control and evaluation of the diabetic animal's response to treatment. The choice of the analytical assay depends on the characteristic and analytical opportunities of the laboratory, as well as the number of serum samples to be analysed.     

Evaluation of fructosamine in dogs and cats with hypo- or hyperproteinaemia, azotaemia, hyperlipidaemia and hyperbilirubinaemia

The influence of various pathological conditions on fructosamine levels in normoglycaemic dogs and cats was investigated. The most frequent and most pronounced deviations were found in animals with hypoproteinaemia, in which fructosamine was significantly lower than in the controls. In 66 per cent of the dogs and 67 per cent of the cats with hypoproteinaemia the levels were below the reference range. In the dogs the concentration of fructosamine was correlated with the level of albumin, but in the cats it was correlated with the level of total protein. Dogs with hyperlipidaemia and azotaemia also had significantly lower levels of fructosamine; 38 per cent of those with hyperlipidaemia and 47 per cent of those with azotaemia had fructosamine levels outside the reference range. No significant changes in fructosamine were detected in dogs or cats with hyperproteinaemia or hyperbilirubinaemia, or in cats with hyperlipidaemia or azotaemia.     

Reliability of history and physical examination findings for assessing control of glycemia in dogs with diabetes mellitus: 53 cases (1995-1998)

OBJECTIVE: To evaluate the reliability of history and physical examination findings for assessing control of glycemia in insulin-treated diabetic dogs. DESIGN: Retrospective study. ANIMALS: 53 insulin-treated dogs with diabetes mellitus. PROCEDURE: Medical records of insulin-treated diabetic dogs from June 1995 to June 1998 were reviewed, and information on owner perception of their dog's response to insulin treatment, physical examination findings, body weight, insulin dosage, and concentrations of food-withheld (i.e., fasting) blood glucose (FBG), mean blood glucose (MBG) during an 8-hour period, blood glycosylated hemoglobin (GHb), and serum fructosamine was obtained. Owner's perception of their dog's response to insulin treatment, physical examination findings, and changes in body weight were used to classify control of glycemia as good or poor for each dog. The FBG, MBG/8 h, blood GHb, and serum fructosamine concentrations were compared between well-controlled and poorly controlled insulin-treated diabetic dogs. RESULTS: Presence or absence of polyuria, polydipsia, polyphagia, lethargy, and weakness were most helpful in classifying control of glycemia. Mean FBG and MBG/8 h concentrations, blood GHb concentrations, and serum fructosamine concentrations were significantly decreased in 25 well-controlled diabetic dogs, compared with 28 poorly controlled diabetic dogs. Most well-controlled diabetic dogs had concentrations of FBG between 100 and 300 mg/dl, MBG/8 h < or = 250 mg/dl, blood GHb < or = 7.5%, and serum fructosamine < or = 525 mumol/L, whereas most poorly controlled diabetic dogs had results that were greater than these values. CONCLUSIONS AND CLINICAL RELEVANCE: Reliance on history, physical examination findings, and changes in body weight are effective for initially assessing control of glycemia in insulin-treated diabetic dogs.     

Evaluation of urine sulfated and nonsulfated bile acids as a diagnostic test for liver disease in dogs

OBJECTIVE: To evaluate 3 methods for measuring urine bile acids (UBA) and compare their diagnostic performance with that of the serum bile acids (SBA) test and other routine screening tests in dogs with hepatic disorders. DESIGN: Prospective study. ANIMALS: 15 healthy dogs, 102 dogs with hepatic disorders, and 9 dogs with clinical signs of hepatic disorders that were found to have nonhepatic disorders. PROCEDURES: Blood and urine samples were collected from sick dogs and healthy dogs for serum biochemical analyses, and determination of concentrations of SBA and UBA. Urine samples were obtained from 15 healthy dogs to establish an upper cutoff value for UBA concentrations. The UBA were measured by use of a quantitative-linked enzymatic colorimetric method. Three analytical modifications were evaluated; 1 quantified only urine sulfated bile acids (USBA), 1 only urine nonsulfated bile acids (UNSBA), and 1 quantified both (USBA plus UNSBA). The UBA values were standardized with the urine creatinine concentration. RESULTS: The UNSBA-to-creatinine ratio and USBA plus UNSBA-to-creatinine ratio tests had the best diagnostic performance of the UBA tests; each had a substantially higher specificity, slightly higher positive predictive value, slightly lower negative predictive value, and lower sensitivity than the SBA test. These UBA-to-creatinine values were positively correlated with SBA values. The USBA-to-creatinine ratio had poor sensitivity, indicating a low rate of bile acid sulfation in dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The UBA can be measured in dogs with sufficient repeatability and accuracy for clinical application. The UNSBA-to-creatinine ratio and USBA plus UNSBA-to-creatinine ratio identified dogs with hepatic disorders nearly as well as the SBA test.     

Association of canine obesity with reduced serum levels of C-reactive protein.

The prevalence of obesity is increasing in dogs as well as in humans. C-reactive protein (CRP) is an important tool for the detection of inflammation and/or early tissue damage and is linked to obesity in humans. The objective of the present study was to determine if serum CRP levels are altered in obese dogs. Fifteen lean (control group) and 16 overweight (obese group) dogs were examined. Blood samples were collected under fasted conditions for serum determination of CRP, glucose, insulin, cholesterol, triglyceride, and fructosamine. Results indicated that obese dogs were insulin resistant because serum insulin and insulin/glucose ratios were higher than in lean dogs (P < or = 0.05). Serum CRP concentrations were lower in obese dogs than in controls (P < or = 0.001). C-reactive protein was negatively correlated with insulin/glucose ratio (R = -0.42) and cholesterol (R = -0.39; P < or = 0.05). Furthermore, levels of cholesterol, triglycerides, and fructosamine were increased in the obese group compared with the control group. Based on these results, it can be postulated that CRP production is inhibited by obesity and insulin resistance in dogs.     

Comparison of Multistix PRO dipsticks with other biochemical assays for determining urine protein (UP), urine creatinine (UC) and UP:UC ratio in dogs and cats

BACKGROUND: Urine protein: urine creatinine (UP:UC) ratio determined from the quantitative measurement of protein and creatinine in a single urine sample is the best feasible assessment of clinically significant proteinuria in dogs and cats. A dipstick that measures urine protein, urine creatinine, and UP:UC ratio has been used in human medicine and could have application for veterinary practice. OBJECTIVE: The objective of this study was to compare the Multistix PRO dipstick (Bayer Corporation, Elkhart, IN, USA) to other biochemical methods for determination of urine protein and creatinine, and UP:UC ratio in canine and feline urine. METHODS: A complete urinalysis, including sulfosalicylic acid (SSA) precipitation, was performed on urine samples submitted to our laboratory between February and April 2003 from 100 dogs and 49 cats. Urine protein and creatinine concentrations were determined by the Multistix PRO dipstick using a Clinitek 50 analyzer (Bayer) and compared with the results of SSA precipitation and quantitative biochemical analysis. The UP:UC ratios from the dipstick results (calculated by the Clinitek 50 and also manually) were compared with those calculated from quantitative values. Pearson product-moment correlation analysis and diagnostic sensitivity and specificity (using quantitative results as the gold standard) were determined. RESULTS: For both canine and feline urine, protein and creatinine concentrations determined by the Multistix PRO correlated closely with quantitative concentrations for protein (dogs r = .78, P = .0001; cats r = .87, P = .0001) and creatinine (dogs r = .78, P = .0001; cats r = .76, P = .0001). The Multistix PRO was more sensitive and less specific than SSA precipitation for diagnosing clinically significant proteinuria. UP:UC ratios obtained by manual calculation of dipstick results correlated best with quantitative UP:UC ratios in dogs, and had higher specificity but lower sensitivity for the diagnosis of proteinuria. In cats, UP:UC ratios determined by the dipstick method did not correlate (r = -.24, P = .0974) with quantitative values. CONCLUSIONS: The Multistix PRO, with manual calculation of UP:UC, may be a good alternative for the diagnosis of clinically significant proteinuria in dogs, but not cats. Dipstick creatinine concentration should be considered as an estimate.     

Fructosamine

Fructosamines are glycated serum proteins that, depending on their life span, reflect glycemic control over the previous 2 to 3 weeks. The nitroblue tetrazolium reduction method adapted to autoanalysis appeared to be a practical means to assay fructosamine quickly, economically, and accurately. The upper limit of the reference range is 374 μmol/L in dogs (95% percentile) and 340 μmol/L in cats (95% percentile). Newly diagnosed diabetic dogs and cats that had not undergone previous insulin therapy had significantly higher fructosamine concentrations than nondiabetic animals. In diabetic dogs that were receiving insulin therapy, the fructosamine test reflected the glycemic state far more accurately than did individual blood glucose measurements. Animals with satisfactory metabolic control revealed fructosamine concentrations within the reference range, whereas fructosamine concentrations above 400 μmol/L indicated insufficient metabolic control. On the basis of fructosamine concentrations, cats with a transitory hyperglycemia and cats with diabetes mellitus were differentiated. The fructosamine test is a valuable parameter for the diagnosis and metabolic control of diabetes mellitus in dogs and cats.     

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 mol/L (median 376 mol/L; reference range 207–340 mol/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.     

Effect of Acute Hyperglycaemia on the Serum Fructosamine and Blood Glycated Haemoglobin Concentrations in Canine Samples

The effect was studied of an acute and non-persistent hyperglycaemia on the serum fructosamine and blood glycated haemoglobin concentrations in canine samples. Five dogs were given glucose solution intravenously and blood samples were taken from each dog before and at 5, 15, 30, 60 and 120 min and 24 h after the infusion. There was an intense hyperglycaemia 5 min after the injection was given, but no statistically significant differences in the serum fructosamine and glycated haemoglobin were observed. It was concluded that an acute and transient hyperglycaemia does not cause significant changes in the glycated haemoglobin and fructosamine concentrations in healthy dogs.     

Haematological and biochemical changes in dogs naturally infected with Angiostrongylus vasorum before and after treatment

Haematological and biochemical parameters were studied prospectively in 48 dogs naturally infected with Angiostrongylus vasorum in a primary care setting. Samples for analysis were obtained when treatment was started and 42days afterwards. Prior to treatment, 21% of affected dogs exhibited eosinophilia, whereas increased total white blood cell (WBC) counts and neutrophilia were observed in only 4.2%. WBC counts and concentrations of neutrophils, eosinophils and monocytes decreased significantly from days 0 to 42, indicating that, even in dogs without elevated absolute blood values, a low grade inflammatory response may be present in dogs with A. vasorum infection. Biochemical changes (especially an increase in serum globulins and a decrease in serum fructosamine) were in agreement with the findings of other studies. The results show that the diagnosis of canine angiostrongylosis should not be excluded based on unremarkable haematological and blood biochemical parameters. They also support our recent finding that a low serum fructosamine concentration may be associated with infection with A. vasorum.     

Diurnal variation in concentrations of various markers of bone metabolism in dogs.

OBJECTIVE: To evaluate diurnal variation in concentrations of selected markers of bone metabolism in dogs. ANIMALS: Ten 3- to 4-year-old ovariectomized Beagles. PROCEDURE: Blood and urine samples were obtained in the morning before dogs were fed (8 AM) and then at 2-hour intervals for 24 hours. This procedure was repeated 2 weeks later. Concentrations of osteocalcin (OC) and carboxy terminal telopeptide of type-I collagen (ICTP) were measured in serum, using a radioimmunoassay; concentrations of hydroxyproline (HYP), pyridinoline (PYD), and deoxypyridinoline (DPD) were analyzed in urine. Hydroxyproline concentration was measured by means of a colorimetric test, whereas PYD and DPD concentrations were quantified by use of high-performance liquid chromatography. RESULTS: In both parts of the study, HYP concentrations increased significantly, compared with values before feeding, until 8 hours after feeding; HYP concentrations then returned to prefeeding values. Concentrations of DPD and PYD decreased from before feeding until 2 PM and then increased until 8 PM. The ICTP concentrations slowly decreased until 4 PM but returned to prefeeding values thereafter. In both parts of the study, concentrations of OC decreased during the day and then increased to reach values similar to those obtained before feeding. CONCLUSIONS: Changes in the concentrations of bone markers were detected throughout the day in the dogs of this study. Increase in HYP concentration most likely was related to feeding. As documented for bone resorption and formation in other species, circadian rhythms were evident for concentrations of DPD, PYD, and OC. Investigators should consider the time of sample collection when measuring these markers.     

Hyperadrenocorticism associated with excessive sex hormone production by an adrenocortical tumor in two dogs.

An 11-year-old spayed female Labrador Retriever and a 9-year-old castrated male miniature Poodle were evaluated because of clinical signs of hyperadrenocorticism. Cortisol testing did not support a diagnosis of hypercortisolemia in either dog; however, imaging studies revealed unilateral adrenal tumors in both dogs. Serum concentrations of 17-hydroxyprogesterone, progesterone, and estradiol were high in both dogs, and androstenedione concentrations were also high in 1 dog. It is suspected that sex hormone secretion by the adrenal tumors in these dogs resulted in clinical signs of hyperadrenocorticism. Clinical signs and hormonal abnormalities resolved in the male dog after surgical resection of the tumor. There was no improvement in clinical signs after treatment with mitotane in the female dog, which died 2 months after diagnosis. Histologic evaluation confirmed the presence of adrenocortical carcinoma in both dogs.     

Acute sterile hemorrhagic cystitis after a single intravenous administration of cyclophosphamide in three dogs.

Three dogs receiving cyclophosphamide IV as part of a combination chemotherapeutic regimen developed macrohematuria, stranguria, and pollakiuria within 24 hours of administration of the first dose of this drug. An 11-year-old spayed mixed-breed dog with an oral squamous cell carcinoma was administered 250 mg of cyclophosphamide/m2 of body surface, whereas a 4-year-old castrated male Gordon Setter was treated with 100 mg of cyclophosphamide/m2 and a 6-year-old male German Shepherd Dog with a cutaneous hemangiosarcoma was administered 140 mg of cyclophosphamide/m2. Aerobic bacterial culture, antimicrobial susceptibility testing, and urinalysis were performed on urine obtained by cystocentesis from all 3 dogs after hematuria was observed. Sterile hemorrhagic cystitis was diagnosed on the basis of large numbers of RBC in the urine, lack of pathogens on bacterial culturing of urine, and clinical signs. Although cyclophosphamide-induced cystitis in dogs has been reported in the literature numerous times, acute episodes developing within 24 hours of administration of the first dose have not been reported in this species with the use of therapeutic doses. Therefore, appropriate precautionary steps should be taken, even when the drug is being administered intermittently.