Calibration in dogs of a subcutaneous miniaturized glucose sensor using a glucose meter for blood glucose determination.

The feasibility of calibrating a glucose sensor by using a wearable glucose meter for blood glucose determination and moderate variations of blood glucose concentration was assessed. Six miniaturized glucose sensors were implanted in the subcutaneous tissue of conscious dogs, and the parameters used for the in vivo calibration of the sensor (sensitivity coefficient and extrapolated current in the absence of glucose) were determined from values of blood glucose and sensor response obtained during glucose infusion. (1) Venous plasma glucose level and venous total blood glucose level were measured simultaneously on the same sample, using a Beckman analyser and a Glucometer II, respectively. The regression between plasma glucose (x) and whole blood glucose (y) was y = 1.12x-0.08 mM (n = 114 values, r = 0.96, p = 0.0001). The error grid analysis indicated that the use of a Glucometer II for blood glucose determination was appropriate in dogs. (2) The in vivo sensitivity coefficients were 0.57 +/- 0.11 nA mM-1 when determined from plasma glucose, and 0.51 +/- 0.07 nA mM-1 when determined from whole blood glucose (t = 1.53, p = 0.18, n.s.). The background currents were 0.88 +/- 0.57 nA when determined from plasma glucose, and 0.63 +/- 0.77 nA when determined from whole blood glucose (t = 0.82, p = 0.45, n.s.). (3) The regression equation of the estimation of the subcutaneous glucose level obtained from the two methods was y = 1.04x + 0.56 mM (n = 171 values, r = 0.98, p = 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)     

Intensive attention improves glycaemic control in insulin-dependent diabetes without further advantage from home blood glucose monitoring: results of a controlled trial.

Forty-six diabetics treated with twice-daily insulin were seen every two weeks for six months in an intensive education programme aided by regular home urine glucose testing. Control was improved with a decrease in 24-hour urinary glucose excretion (median 138 mmol/24 h (24.8 g/24 h) falling to 70 mmol/24 h (12.6 g/24 h); p less than 0.002), glycosylated haemoglobin concentration (mean 11.4 +/- SD 2.3% falling to 10.4 +/- 1.5%; p less than 0.001), and Diastix score (median 3.0 falling to 1.3; p less than 0.001). There was no reported increase in hypoglycaemia. Thirty-eight of the diabetics proceeded to a nine-month randomised cross-over study of the effect on blood glucose control of monitoring urinary glucose or blood glucose measured visually or by a reflectance meter using appropriate reagent strips. No further improvement in control was observed after home blood glucose monitoring. Nevertheless, 29 out of 37 patients preferred blood to urine glucose monitoring. During both the education and cross-over studies there was evidence of an initial improvement in control followed by deterioration. This was independent of the monitoring method used in the cross-over period and may have been due to waning enthusiasm. Despite patient enthusiasm and other reports to the contrary, home blood glucose monitoring offered no improvement in control over intensive attention and conventional urine glucose monitoring.     

Precision and costs of techniques for self-monitoring of serum glucose levels.

The poor correlation between serum and urine glucose measurements has led to the development of new techniques for monitoring the blood glucose level in diabetic patients. Either a nurse or the patient can perform these tests, which involve spreading a single drop of blood onto a reagent strip. A colour change that is proportional to the serum glucose level can be read visually or with a reflectance meter. Evaluated against simultaneous serum glucose levels determined by the hospital biochemistry laboratory, those of the new techniques employing reflectance meters all showed excellent correlation (r2 = 0.85 to 0.96). Reagent strips used without meters showed poorer correlation (r2 = 0.69 to 0.90). The instruction given to the patients and one nurse enabled them to obtain more accurate results with one of the meters than nurses not specially trained (r2 = 0.94 and 0.92 v. 0.85 respectively). The mean cost per glucose determination with the new techniques was 75, compared with +1.45 for the laboratory determinations done with automated equipment. It was concluded that the new techniques compared well with the reference method, particularly when reflectance meters were used, and that they were easily applied by the patient, as well as the medical staff, at a reasonable cost.     

Validation of a glucose meter at an intensive care unit

IntroductionHyperglycemic patients admitted to Intensive care units (ICUs) have higher morbidity and mortality than normoglycemic patients. Blood glucose levels of ICU patients are usually measured with a glucose meter. The aim of this study was to evaluate a glucose meter (StatStrip, Nova Biomedical) to assess its agreement with the standard laboratory method for testing glucose.Material and methodsEighty-nine different samples were collected from patients (76.4% men and 23.6% women) admitted to an ICU from September to December 2010. Each blood sample was collected into two tubes, a lithium heparin tube and an EDTA tube. The total blood aliquot was used to measure glycemia using the glucose meter. The lithium heparin tube was processed at the same time for measuring plasma glucose (Cobas 6000 Analyzer, Roche Diagnostics, SA). Agreement between the two methods was assessed according to the EP-9-A2 Clinical Laboratory Standards Institute guideline.ResultsMean whole blood glucose level measured by the glucose meter was 126.53 + 49.28 mg/dL (range, 33.5-431 mg/dL), while mean plasma glucose value measured by the laboratory reference method was 138.13 + 78.6 mg/dL (range, 43-451 mg/dL). Correlation coefficient was 0.99, with a 95% confidence interval of 0.98 to 0.99. Coefficient of determination (R2) was 0.97, and intraclass correlation coefficient was 0.99 with a 95% CI of 0.98 to 0.99.ConclusionsThe tested glucose meter (StatStrip) shows a good linear association, precision, and accuracy when compared to the laboratory reference method. This device is adequate for glucose monitoring.     

Urine glucose levels are disordered before blood glucose level increase was observed in Zucker diabetic fatty rats

Many patients with diabetes are not diagnosed at all or are diagnosed too late to be effectively treated, resulting in nonspecific symptoms and a long period of incubation of the disease. Pre-diabetes is an early warning signal of diabetes, and the change of urine glucose in this period has been ignored even though urine has long been related with diabetes. In this study, Zucker diabetic fatty (ZDF) rats were used to test if there were changes in urine glucose before blood glucose increases. Six 8-week-old male ZDF rats (fa/fa) and Zucker lean (ZL) rats (fa/+) were fed with Purina 5008 high-fat diet and tested for fasting blood glucose and urine glucose. After 12 weeks of feeding, the urine glucose values of the ZL rats were normal (0–10 mmol L~(-1)), but the values of the ZDF model rats increased 10 weeks before their blood glucose levels elevated. The urine glucose values of the ZDF model rats showed a state of disorder that was frequently elevated (10 mmol L~(-1)) and occasionally normal (0–10 mmol L~(-1)). This finding may provide an easy early screening for diabetes by long-term monitoring of urine glucose levels: pre-diabetes may be revealed by frequently disordered urine glucose levels over a period.     

Quantification of hepatic carbohydrate metabolism in conscious mice using serial blood and urine spots

In vivo studies of hepatic carbohydrate metabolism in (genetically modified) conscious mice are hampered by limitations of blood and urine sample sizes. We developed and validated methods to quantify stable isotope dilution and incorporation in small blood and urine samples spotted onto filter paper. Blood glucose and urinary paracetamol-glucuronic acid were extracted from filter paper spots reproducibly and with high yield. Fractional isotopomer distributions of glucose and paracetamol-glucuronic acid when extracted from filter paper spots were almost identical to those isolated from the original body fluids. Rates of infusion of labeled compounds could be adjusted without perturbing hepatic glucose metabolism. This approach was used in mice to find the optimal metabolic condition for the study of hepatic carbohydrate metabolism. In fed mice, no isotopic steady state was observed during a 6-h label-infusion experiment. In 9-h-fasted mice, isotopic steady state was reached after 3 h of label infusion and important parameters in hepatic glucose metabolism could be calculated. The rate of de novo glucose-6-phosphate synthesis was 143 +/- 17 micromol kg(-1) min(-1) and partitioning to plasma glucose was 79.0 +/- 5.2%. In 24-h-fasted mice, abrupt changes were noticed in whole body and in hepatic glucose metabolism at the end of the experiment.     

Impaired glucose metabolism in hypertensive patients

We have studied glucose tolerance under carefully controlled conditions in 79 patients with arterial hypertension. The results show that, in patients with arterial hypertension but without clinical diabetes mellitus, the glucose tolerance was abnormal in 77.3% and normal in 22.3%. The corresponding figure in the control group of normotensive subjects was 0%. In each test the responses to glucose administration were analyzed by plotting the logarithm of the blood glucose concentration against time. For the points between 60 and 120 min, corresponding to the periods following glucose administration, a linear relationship was obtained and showed a decline at an exponential rate, as noted by other observers. An estimate of the volume of distribution of glucose was obtained as follows. Values observed in hypertensives with a pathological percent fall in blood glucose per minute (Kg) were 29.8 +/- 12.0 (mean +/- SD) liters and those in normal subjects with normal Kg values had a mean of 14.35 +/- 2.98, the difference being highly significant (p less than 0.0001). The results of the theoretical glucose concentration are also presented. Those obtained from subjects with normal Kg values (359.0 +/- 58.4 mg/dl) are significantly higher than in subjects with pathological Kg values (257.6 +/- 51.3 mg/dl; p less than 0.0001). All patients with either pathological or normal Kg values had normal glucose concentration levels, fasting blood sugar and no glucose in the urine specimen. The difference between pathological Kg values (107.0 +/- 25.8 mg/dl) and normal Kg values (90.6 +/- 13.0 mg/dl) was not found to be statistically different (p greater than 0.05). The distribution and means of glucose half time in controls with normal Kg values and hypertensives with pathological Kg values were: 63.5 +/- 11.5 and 137.8 +/- 48.1 min, respectively. The difference between normal and pathological Kg values being statistically significant at a confidence level above 99.5%. We also studied the free glucose pool at zero time. A significantly higher level was found in hypertensives with pathological Kg values, again indicating an impairment in glucose metabolism in this group: 90.6 +/- 26.5 vs. 65.0 +/- 5.4 g (p less than 0.0001). Another study showed an estimate of the mean cellular glucose uptake (MCUg) per minute and per kilogram body weight. The MCUg following glucose loading decreased considerably in hypertensives with pathological Kg values. The percentage reduction ranged between 50 and 55% hypertensives with pathological Kg values 4.1 +/- 0.8, and normotensives with normal Kg values, 8.0 +/- 0.6 (p less than 0.0001).(ABSTRACT TRUNCATED AT 400 WORDS)     

Heparanase levels are elevated in the urine and plasma of type 2 diabetes patients and associate with blood glucose levels

Heparanase is an endoglycosidase that specifically cleaves heparan sulfate side chains of heparan sulfate proteoglycans. Utilizing an ELISA method capable of detection and quantification of heparanase, we examined heparanase levels in the plasma and urine of a cohort of 29 patients diagnosed with type 2 diabetes mellitus (T2DM), 14 T2DM patients who underwent kidney transplantation, and 47 healthy volunteers. We provide evidence that heparanase levels in the urine of T2DM patients are markedly elevated compared to healthy controls (1162 ± 181 vs. 156 ± 29.6 pg/ml for T2DM and healthy controls, respectively), increase that is statistically highly significant (P<0.0001). Notably, heparanase levels were appreciably decreased in the urine of T2DM patients who underwent kidney transplantation, albeit remained still higher than healthy individuals (P<0.0001). Increased heparanase levels were also found in the plasma of T2DM patients. Importantly, urine heparanase was associated with elevated blood glucose levels, implying that glucose mediates heparanase upregulation and secretion into the urine and blood. Utilizing an in vitro system, we show that insulin stimulates heparanase secretion by kidney 293 cells, and even higher secretion is observed when insulin is added to cells maintained under high glucose conditions. These results provide evidence for a significant involvement of heparanase in diabetic complications.     

Limitation of glucose oxidase method of glucose estimation in jaundiced neonates

The most widely used method for estimation of plasma glucose is that adopted by Trinder's using glucose oxidase-peroxidase (GOD-POD) system. This method gives much lower blood glucose values with blood samples of neonatal jaundice (plasma bilirubin level > 10 mg/dL) of age 10 +/- 5 daysthan with samples of neonates of the same age group without jaundice or older children suffering from other diseases like acute respiratory distress, septicemia.     

Effects of training on glucose metabolism of gluconeogenesis-inhibited short-term-fasted rats

To evaluate the effects of endurance training on gluconeogenesis and blood glucose homeostasis, trained as well as untrained short-term-fasted rats were injected with mercaptopicolinic acid (MPA), a gluconeogenic inhibitor, or the injection vehicle. Glucose kinetics were assessed by primed-continuous venous infusion of [U-14C]- and [6-3H]glucose at rest and during submaximal exercise at 13.4 m/min on level grade. Arterial blood was sampled for the determination of blood glucose and lactate concentrations and specific activities. In resting untrained sham-injected rats, blood glucose and lactate were 7.6 +/- 0.2 and 1.3 +/- 0.1 mM, respectively; glucose rate of appearance (Ra) was 71.1 +/- 12.1 mumol.kg-1.min-1. MPA treatment lowered blood glucose, raised lactate, and decreased glucose Ra. Trained animals had significantly higher glucose Ra at rest and during exercise. At rest, trained MPA-treated rats had lower blood glucose, higher blood lactate, and similar glucose Ra and disappearance rates (Rd) than trained sham-injected animals. Exercising sham-injected untrained animals had increased blood glucose and glucose Ra compared with rest. Exercising trained sham-injected rats had increased blood glucose and glucose Ra and Rd but no change in blood lactate compared with untrained sham-injected animals. In the trained animals during exercise, MPA treatment increased blood lactate and decreased blood glucose and glucose Ra and Rd. There was no measurable glucose recycling in trained or untrained MPA-treated animals either at rest or during submaximal exercise. There was no difference in running time to exhaustion between trained and untrained MPA-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS)     

Utilization of glucose during exercise in relation of meal-timing

The effect of glucose administration was studied on its utilization during exercise carried out in the hours 500-700, 1100-1300, 1700-1900, 2300-100. The control group comprised animals at rest which had one or two glucose loads. Circadian variability of blood glucose level was observed in response to glycaemic stimulation in control animals. In the animals during exercise the circadian changes of glucose level depended on the time after glucose administration and the duration of exercise. Glucose utilization during exercise was not identical at various times of the 24-hour period. The greatest fall of blood glucose was observed at 1800 after one as well as after two glucose loads. Glucose administration after one hour of exercise prevented hypoglycaemia development.     

Mouse urine collection using clear plastic wrap

Qualitative urinalysis using Multistix reagent strips for the detection of urinary pH, protein, glucose, bilirubin, blood, ketone, urobilinogen and creatinine can be carried out with a few drops of mouse urine. The use of metabolic cages is not practical for such qualitative studies particularly when several animals are involved. Here we describe two different methods for collecting pure mouse urine. The single animal method (SAM) involves allowing a single mouse to urinate on Glad cling wrap outside of the animal cage. The multiple animal method (MAM) involves partitioning seven mice into seven different make-shift compartments laid out on top of the cling wrap and allowing them to urinate. The voided urine, in each case, is then aspirated into micro-centrifuge tubes using a Pipetman. Without coercion pure urine was obtained as early as 12 s. Volumes in the range of 10-250 microl were obtained. Modifications of the SAM could prove useful for rat or mouse urine collection under conditions of microgravity.     

Self monitoring of glucose by people with diabetes: evidence based practice.

The inappropriate use of self monitoring of glucose is wasteful of NHS resources and can cause psychological harm. Although a few patients find that self monitoring enables them to understand and take control of their diabetes, many people with diabetes are performing inaccurate or unnecessary tests. There is no convincing evidence that self monitoring improves glycaemic control, nor that blood testing is necessarily better than urine testing. It may be appropriate for some patients not to monitor their own glucose but to rely instead on regular laboratory estimations of glycaemic control. Glucose self monitoring should be performed only when it serves an identified purpose. It is widely assumed that glucose self monitoring, preferably of blood glucose concentrations, is desirable or even essential for everyone with diabetes. It is common for patients who have previously tested their urine, or have done no glucose monitoring at home, to be taught to measure their blood glucose when they are admitted to hospital. In the community too, patients are often encouraged to monitor their blood glucose, and newly diagnosed patients of all ages are usually taught to measure their blood glucose concentrations. Self monitoring can sometimes be useful, but evidence is mounting that its indiscriminate use is of questionable value. In 1995, Pounds 42.6 million was spent on home monitoring of glucose in the United Kingdom (Intercontinental Medical Statistics, personal communication). Is this enormous cost justified? Is blood testing necessarily better than urine testing? Is glucose self monitoring always necessary, or is it sometimes a waste of time and money? Are recommendations for self monitoring based on sound evidence?     

Evaluation of the quality of bedside monitoring of the blood glucose level in a teaching hospital.

We evaluated the precision and accuracy of monitoring of the blood glucose level in the laboratory and at the bedside with one of four glucose meters by an experienced operator and by 39 nurses in a teaching hospital. For precision studies aqueous quality control materials were used. A total of 85 blood samples were tested. The precision of the glucose meters (expressed as the coefficient of variation [CV]) in the hands of the experienced operator ranged from 6.7% to 11.1%. The correlation between the values obtained by the experienced operator and the reference values obtained in the laboratory was high (0.95 to 0.98). The precision of the values obtained by the nurses using the meters ranged from 13.7% on medical wards to 45.7% in the intensive care unit (ICU). The correlation between these values and those obtained in the central laboratory ranged from 0.72 to 0.82. Twenty-four percent of the glucose values determined on medical wards and 62% of those determined in the ICU deviated from the reference value by at least 20%. Of the 85 patients 12 (14%) would have received different insulin dosages had the reference value been available at the same time as the glucose meter reading: in 3 of the patients the discrepancy was 6 units of insulin or greater. Continuous quality control of bedside monitoring of the blood glucose level is needed. In addition, personnel who use glucose meters should receive adequate training.     

Insulin and glucose in neonatal calves after peroral insulin and intravenous glucose administration.

Effects of peroral insulin on plasma concentrations of immunoreactive insulin (IRI) and glucose in newborn calves were studied. Bovine insulin was administered in amounts of 0.5 mg/kg body weight immediately preceding first colostrum. Thereafter, neither an increased IRI response nor a lowering of blood glucose level were observed, indicating that insulin was either not absorbed from the intestine or possibly retained in the liver. Feeding of whole milk was followed by a higher rise in IRI and glucose concentrations than feeding of colostrum after po insulin. However, when compared with 1-3-month old calves, IRI responses to feeding and to iv infused glucose were markedly smaller on the first and second day of life respectively, while glucose responses were similar. This indicates that insulin secretory mechanisms are not fully developed in the newborn calf.     

Interconnection between parameters of motor activity and blood glucose concentration in newborn rats at starvation and under glucose load conditions

The study is carried out on the one-, 3-, 5-, 7, and l0-day old free moving or loosely fixed rat pups (P1–10). In satiated and submitted to the 24-h starvation rat pups, parameters of spontaneous periodical motor activity (SPMA) and the blood glucose content were determined. The total glucose level in hungry rat pups was, on average, 1.5–2.5 times lower than in satiated animals. Administration of glucose to hungry rat pups increased 6–11 times its concentration in blood as compared with the animals not obtaining glucose. The glucose administration to the satiated rat pups led to a rise of its level in blood from 2 to 5 times as compared with intact animals, which was 2–3 times less than in the case of hungry animals. Analysis of pattern of motor activity recorded under conditions of the glucose deficit caused by the 24-h starvation of rat pups did not reveal significant changes of ratio of rhythmical components. Introduction of glucose to hungry and to the lesser degree to satiated rat pups led to potentiation of the minute rhythm of activity in all age groups. The exception was the first day after birth when the glucose administration to satiated rat pups promoted an enhancement of the decasecond rhythm and a decrease of the total level of motor activity. Comparison of ontogenetic dynamics of the SPMA parameters and the glucose content in blood of hungry rat pups revealed the clearly expressed regularity absent in the satiated animals: the glucose level in blood was higher during activity than in the state of rest. The performed study has shown that intensity, duration, and, to a degree, pattern of SPMA in the newborn rat pups depend on the level of satiety, and can be significantly changed in the artificially produced hypo- or hyperglycemia. The existing ontogenetic fluctuations in the character of reaction at performance of the glucose tolerance test can be connected both with morphofunctional maturation of the motor system and with immaturity of various chains of carbohydrate metabolism.     

Single-mouse urine collection and pH monitoring system.

A glass cage with minimal surface area was designed and used to house mice for 24-hour urine collections. An experiment was performed with a radio-labeled compound excreted in the urine to assess the collection efficiency of the cage. In this experiment 74.2 +/- 6.5% of the excreted radioactivity was recovered in the urine, with 25.8 +/- 6.5% found adhering to the cage surfaces. When a flow-through pH electrode, meter, and recorder were attached, the system provided a continuous pH versus time urination record.     

Nanogold-plasmon-resonance-based glucose sensing

Noble metal nanoparticles are well known for their strong interactions with light through the resonant excitations of the collective oscillations of the conduction electrons on the particles, the so-called surface plasmon resonances. The close proximity of two nanoparticles is known to result in a red-shifted resonance wavelength peak, due to near-field coupling. We have subsequently employed this phenomenon and developed a new approach to glucose sensing, which is based on the aggregation and disassociation of 20-nm gold particles and the changes in plasmon absorption induced by the presence of glucose. High-molecular-weight dextran-coated nanoparticles are aggregated with concanavalin A (Con A), which results in a significant shift and broadening of the gold plasmon absorption. The addition of glucose competitively binds to Con A, reducing gold nanoparticle aggregation and therefore the plasmon absorption when monitored at a near-red arbitrary wavelength. We have optimized our plasmonic-type glucose nanosensors with regard to particle stability, pH effects, the dynamic range for glucose sensing, and the observation wavelength to be compatible with clinical glucose requirements and measurements. In addition, by modifying the amount of dextran or Con A used in nanoparticle fabrication, we can to some extent tune the glucose response range, which means that a single sensing platform could potentially be used to monitor microM --> mM glucose levels in many physiological fluids, such as tears, blood, and urine, where the glucose concentrations are significantly different.     

Quantitation of doxapram in blood,plasma and urine

Methods for the quantitation of doxapram in blood, plasma and urine have been developed. Following extraction, gas—liquid chromatography was used to separate doxapram from basic metabolites. Doxapram was detected by mass spectrometry for blood and plasma assays, and by flame ionisation for urine assays. The limit of reliable quantitation in blood and plasma was 10 ng and in urine 500 ng, the coefficients of variation being 6.37%, 1.72% and 2.31% respectively. To illustrate the clinical applicability of the assay methods, plasma, blood and urine levels were monitored in a premature newborn following an intravenous infusion of doxapram.     

Determination of quinalphos in blood and urine by direct solid-phase microextraction combined with gas chromatography-mass spectrometry

A new method based on direct solid-phase microextraction (DI-SPME) followed by gas chromatography-mass spectrometry was developed for the purpose of determining quinalphos in blood and urine. Two types of coated fibre have been assayed and compared: carbowax/divinylbenzene (CW/DVB 65 microm) and polydimethylsiloxane (PDMS 100 microm). The main parameters affecting the SPME process such as temperature, salt addition, pH, stirring and adsorption/desorption time profiles were optimized to enhance the sensitivity of the procedure. The method was developed using only 100 microL of blood and urine. Limits of detection of the method for blood and urine matrices were, respectively, 10 and 2 ng/mL. Linearity was established over concentration ranges from 0.05 to 50 microg/mL for blood, and 0.01 to 50 microg/mL for urine, with regression coefficients ranging between 0.9991 and 0.9999. Intra- and interday precision values were less than 13%, and accuracy was within +/-15% of the nominal concentration for all studied levels in both matrices. Absolute recoveries were 14 and 26% for blood and urine, respectively.