Plasma nitrotyrosine in reversible myocardial ischaemia.

BACKGROUND: Nitric oxide (NO) plays a vital role in vascular homeostasis and in the pathophysiology of coronary heart disease. Its metabolites, nitrite and nitrate, have vasculoprotective properties, whereas peroxynitrite, an oxidant metabolite of NO, is cytotoxic and can aggravate myocardial damage during ischaemic reperfusion injury. Peroxynitrite nitrates free and protein bound tyrosine residues to produce nitrotyrosine. The measurement of nitrotyrosine provides an indirect estimation of plasma peroxynitrite concentrations. AIMS: To measure plasma nitrotyrosine concentrations to see whether peroxynitrite could contribute to myocardial dysfunction during myocardial ischaemia induced by an exercise tolerance test (ETT). MATERIALS/METHODS: Plasma free nitrotyrosine concentrations were compared before and after exercise in 29 subjects with a positive ETT and 34 subjects with a negative ETT. RESULTS: Plasma nitrotyrosine concentrations were similar in patients with exercise induced myocardial ischaemia and controls. CONCLUSION: Peroxynitrite does not contribute to the myocardial dysfunction in reversible myocardial ischaemia.

Hypertension-based clinical risk strategies for detecting microalbuminuria in diabetes.

BACKGROUND: Microalbuminuria screening to identify patients at risk of diabetic nephropathy is widely accepted. AIM: To investigate whether blood-pressure-based strategies can identify such patients without the need for microalbuminuria testing. METHODS: Spot urine for albumin/creatinine ratios was performed in all patients over an 18-month period. The performance of four combinations of clinical models, based on existing triggers for anti-hypertensive intervention (prior use and/or existing systolic BP exceeding 140 or 160 mmHg and/or dipstick proteinuria exceeding 1+ or 2+) was evaluated at microalbuminuria thresholds of 3.5 and 10 mg/mmol. The models were ranked 1 to 4, based on their escalating relative strengths in predicting need for intervention. RESULTS: Of 3748 patients, 1257 (34%) or 739 (20%) exceeded microalbuminuria thresholds of 3.5 or 10 mg/mmol. All four models predicted microalbuminuria risk (areas under ROC curves 0.60-0.77, all p < 0.001). The models (1-4) identified 2220, 2465, 2803 or 2937 for intervention, respectively, irrespective of microalbuminuria status, and missed 368, 232, 194 or 126 at 3.5 mg/mmol and 164, 87, 81 or 45 at 10 mg/mmol. DISCUSSION: Clinical models using routinely measured parameters reduced the target population for microalbuminuria screening by 60-80%, missing 3-10% of patients with albumin/creatinine ratios exceeding 3.5 mg/mmol or 1-4% of those exceeding 10 mg/mmol.

Cardiovascular effects of platelet-activating factor.

Platelet-activating factor (PAF) is a unique biologically active phospholipid that is synthesized by many cells and tissues and has actions far more diverse than just platelet activation. PAF is probably a mediator of the normal inflammatory and thrombotic responses. Inappropriate or excessive production of PAF can lead to vascular injury, particularly via its proinflammatory effects, and marked hemodynamic derangements. There is evidence that PAF may play a role in clinical syndromes of shock, infarction, and reperfusion injury. Antagonists of the PAF receptor have been developed and have therapeutic potential in these common life-threatening disorders.

Quantitative ultrasonic imaging: tissue characterization and instantaneous quantification of cardiac function.

Quantitative myocardial tissue characterization is being developed to complement and expand conventional echocardiography by delineating the physical state of myocardium under diverse pathophysiologic conditions. Real-time quantitative integrated backscatter imaging has already been applied to patients with ischemic heart disease, hypertrophic cardiomyopathy, and cardiac allograft rejection in clinical investigations performed in the United States, Europe, and Japan. A recently introduced modification of imaging processing algorithms employed for characterization of tissue facilitates automatic detection of endocardial-blood interfaces and on-line quantification of ventricular size and function. Further progress and anticipated developments in quantitative ultrasonic imaging will undoubtedly augment the clinical applications of tissue characterizations based on myocardial integrated backscatter for improved diagnosis, elucidation of pathophysiology, and assessment of cardiac function.

Automated, on-line quantification of left ventricular dimensions and function by echocardiography with backscatter imaging and lateral gain compensation.

To provide on-line quantification of left ventricular cavity dimensions and function by echocardiography 60 control subjects and 10 patients with cardiac dysfunction were studied. A novel, ultrasound imaging system was used which was developed to detect and track, in real time, ventricular endocardial blood boundaries based on quantitative assessment of acoustic properties of tissue. In addition, lateral gain compensation, a robust and novel image enhancement procedure, was used to provide instantaneous measurement and display of cavity areas and functional indexes on a beat-by-beat basis within regions of interest drawn around the blood pool cavity. In control subjects, short-axis end-diastolic area averaged 13.1 +/- 3.7 cm2 (SD), end-systolic area 5.9 +/- 2.7 cm2, and fractional area change 55.6 +/- 11.2%. Apical views yielded corresponding values of 23.8 +/- 4.5 cm2, 15.5 +/- 3.4 cm2 and 34.7 +/- 7.8%. Instantaneous peak rate of cavity area change approximated 50 cm2/s in systole and 60 cm2/s in diastole in each view. Serial measurements of area and functional index were reproducible over intervals of 2 to 3 weeks. Patients with dilated ventricles exhibited average apical view area values of 49.1 +/- 6.1 cm2 and 43.1 +/- 4.9 cm2 in diastole and systole with a fractional area change of 12.2 +/- 3.0%. Thus, results with on-line echocardiographic backscatter imaging-assisted automated edge detection are reproducible and capable of delineating cardiac dysfunction conveniently, promptly and serially at the bedside.

Hypertension in diabetes: is there a place for age-adjusted centile cut-offs in those aged <50 years?

BACKGROUND: Conventional cut-offs for hypertension are arbitrary, and vascular complications can occur below these values, particularly in diabetes. AIM: To evaluate systolic blood pressure (SBP) distribution in diabetes and control populations, comparing hypertension prevalence using either conventional cut-offs (140 and 160 mmHg) or age-adjusted centile (75(th) and 90(th)) cut-offs derived from the control population. METHODS: We compared 2521 individuals with diabetes to 5809 controls, after excluding those on anti-hypertensives and with established vascular disease in both groups. RESULTS: Diabetic individuals were older (mean +/- SD 56 +/- 16 vs. 43 +/- 16years, diabetes vs. controls), and had higher BMI (29 +/- 5 vs. 24 +/- 4 kg /cm(2)) and SBP (145 +/- 23 vs. 131 +/- 18 mmHg, all p < 0.001). These factors were adjusted for in subsequent analysis. SBP rose with age and was also significantly higher in the diabetes group. In diabetes, conventional cut-offs indicated less hypertension in those aged < 50 years, compared to age-adjusted centile cut-offs. In age bands 18-29, 30-39, 40-49, 50-59, 60-69 and >70 years of the diabetes group, 24%, 33%, 43%, 62%, 70% and 74%, respectively exceeded 140 mmHg, compared to 35%, 44%, 43%, 45%, 40% and 27% exceeding the control-derived 75(th) centile value. DISCUSSION: The use of control-derived age-adjusted cut-offs may provide an alternative approach to define hypertension in diabetes that may be of particular relevance to younger patients, although this would require validation against outcomes.

Recurrent left ventricular false aneurysm.

A recurrent left ventricular false aneurysm 5 years after patch repair and causing progressive congestive heart failure was readily diagnosed by echocardiography and heart catheterization. Its substrate was suture dehiscence of undetermined origin. Urgent repair was successful. The long-term prognosis is guarded.

Transmural variation of myocardial attenuation and its potential effect on contrast-mediated estimates of regional myocardial perfusion.

Promising technical developments suggest that it may be feasible to use contrast echocardiography to estimate regional myocardial perfusion. Although the optimal approach has not yet been determined, the use of a nonlinear (harmonic) response of the contrast agent is common to several recent advances. The purpose of this article is to delineate the relation between the anisotropic (angle-dependent) ultrasonic attenuation of the myocardium through which the sound wave has propagated and the regional, nonlinear response of the contrast agent. Apparent perfusion will be modulated by this regionally varying, path-dependent attenuation, which is determined by the local angle between the propagating sound wave and the myofiber orientation. We illustrate the potential magnitude of the effect of myocardial anisotropy for the apical 4-chamber view by examining propagation along the septum and the lateral wall. We present experimentally measured values of the attenuation of excised sheep myocardium, showing statistically significant differences in the attenuation in the mid wall compared with that in symmetrical zones to the left and right of the mid wall, reflecting the well-known myofiber orientations in these 3 regions. The nonlinear (harmonic) response of a contrast agent depends on the local pressure amplitude, which for a given mechanical index is determined by the attenuation accumulated along the path to the point where the regional perfusion is estimated.

Cyclic variation of integrated backscatter: dependence of time delay on the echocardiographic view used and the myocardial segment analyzed.

To determine the influence of myocardial anisotropy in ultrasonic tissue characterization, we measured the time delay (and magnitude) of the cyclic variation of myocardial integrated backscatter from specific segments visualized in the 4 standard transthoracic echocardiographic views. The cyclic variation data in 10 myocardial regions were obtained from analyses of 2-dimensional integrated backscatter images from 23 healthy subjects. Resultant values (mean +/- SD) for the time delay were as follows: parasternal long-axis view: 1.08 +/- 0.17 (septum) and 1.00 +/- 0.14 (posterior wall); parasternal short-axis view: 1.03 +/- 0.16 (anterior septum), 1.03 +/- 0.14 (posterior wall), 2.22 +/- 0.71 (lateral wall), and 1.65 +/- 0.66 (posterior septum); apical 4-chamber view: 1.08 +/- 0.31 (septum) and 2.20 +/- 0.79 (lateral wall); and apical 2-chamber view: 1.68 +/- 0.62 (inferior wall) and 2.04 +/- 0.72 (anterior wall). Hence, results of this study indicate that myocardial ultrasonic characterization that uses the cyclic variation is influenced by the echocardiographic view and the specific segment of the left ventricle.

Effects of myocardial fiber orientation in echocardiography: quantitative measurements and computer simulation of the regional dependence of backscattered ultrasound in the parasternal short-axis view.

We measured the regional disparity in backscattered ultrasound by means of obtaining integrated backscatter images of 10 healthy subjects and placing a region of interest in 18 distinct positions. A computer model simulating the short-axis view was implemented on the basis of previously measured values for the anisotropic ultrasonic properties of myocardium. Measurements showed that the integrated backscatter value was greatest for the anterior septum and decreased by 15.9 +/- 3.5 dB for the lateral wall and 17.7 +/- 3.5 dB for the inferior septum. The value in the posterior wall was 8.1 +/- 3.8 dB below the value for the anterior septum. The regional variation of backscatter predicted with the simulation correlated well with the clinical measurements. These results suggested that analyses based on measurements of backscatter may require compensation for the inherent anisotropic properties of myocardium.

Effects of tissue anisotropy and contrast acoustic properties on myocardial scattering in contrast echocardiography.

In this study we explored the potential effects that tissue anisotropy, in conjunction with the acoustic properties of contrast, may have on quantitative measurements of myocardial perfusion with the use of ultrasonic contrast agents. We used a computer simulation of the parasternal short-axis view, based on previously measured values for the anisotropy of backscatter and attenuation of myocardium, to predict the backscattered energy from 18 specific regions within the heart before and after myocardial contrast perfusion. Results demonstrated a regional variation of contrast enhancement in the short-axis view and variations caused by incremental increases in contrast level for specific myocardial regions. Thus quantitative assessment of myocardial perfusion with contrast echocardiography is influenced by the anisotropic properties of the myocardium, and the resulting postcontrast image will depend on the interaction between tissue properties and contrast acoustic properties. The degree of myocardial enhancement caused by the presence of contrast may depend on the spatial position of the specific region investigated with respect to the transducer and the amount of contrast in the myocardium.

Comparison of integrated backscatter values obtained with acoustic densitometry with values derived from spectral analysis of digitized signals from a clinical imaging system.

Time-domain-based integrated backscatter values obtained with the use of acoustic densitometry (AD) were compared with values determined from a spectral-based analysis of the radio-frequency (RF) signals with a modified Hewlett-Packard Sonos 1500 imaging system. Integrated backscatter images of five specimens of bovine tendon were acquired in the AD acquisition mode, and the corresponding signals related to the backscattered RF were digitized for each angle of insonification as the specimens were rotated in 10-degree increments. The integrated backscatter images were analyzed with the AD analysis package, and the corresponding values determined from the RF power spectra were obtained from the digitized ultrasonic signals. Good agreement was found between the two methods over the entire range of measured values. The mean anisotropy in the measured integrated backscatter (mean +/- standard error) was found to be 27 +/- 2 dB for time-domain-based analysis and 25 +/- 2 dB for RF spectral-based analysis.

On the transportability of linear discriminant functions.

Different methods can give different results for the same analyte, and inconsistent answers may be obtained if such results are used in linear discriminant functions. This is an important factor to consider when transporting linear discriminant functions from one laboratory to another; the example of alkaline phosphatase activity and the differential diagnosis of hypercalcaemia has recently been given. This study shows that results obtained by the method recommended by the Scandinavian Committee on Enzymes for alkaline phosphatase may be used in the calculation of the discriminant functions used for the differential diagnosis of hypercalcaemia and considers other factors that may affect the discrimination. It is concluded that, with care, linear discriminant functions may be transported from one laboratory to another.

Effect of a protease inhibitor on in vitro stability of intact parathyroid hormone.

BACKGROUND: We investigated whether increased protease activity explains the increased in vitro degradation of intact parathyroid hormone (iPTH) observed in serum when compared to EDTA plasma. METHODS: Pre-dialysis blood samples for iPTH were taken from 11 patients with chronic renal failure and collected into plain glass tubes, tubes containing 200 KIU/mL aprotinin (a protease inhibitor) and EDTA tubes. All sample aliquots were separated at 20 min, 1 h, 2 h, 4 h, 8 h and 24 h post collection. RESULTS: Over 24 h, iPTH concentrations remained unchanged in EDTA tubes. iPTH concentrations were significantly lower in both plain tubes (P < 0.01) and aprotinin tubes (P < 0.001) at 24 h when compared to the baseline sample (20 min). At 24 h, iPTH concentrations in EDTA tubes were higher than in plain tubes (P < 0.001) and aprotinin tubes (P < 0.01). The addition of aprotinin to plain tubes significantly reduced the degradation of iPTH (P < 0.05) at 24 h. CONCLUSION: Aprotinin significantly reduces the in vitro degradation of iPTH in plain tubes at 24 h from 24.7% to 9.6%. We suggest that increased protease activity contributes to the decline in serum iPTH over time. As this is observed in serum and not plasma it suggests that the increased protease activity may be due to the clotting process.

Hemodialysis-induced hypoglycemia in diabetic patients.

BACKGROUND: The authors have previously reported asymptomatic hypoglycemia in non-diabetic patients on hemodialysis. The present study was designed to assess the effect of hemodialysis on plasma glucose, glucagon, cortisol and catecholamines in diabetic patients. METHODS: Eighteen diabetic patients with chronic renal failure were hemodialyzed using a glucose-free dialysis fluid. They did not take any medication prior to dialysis and were asked not to eat during the first hour on hemodialysis. Blood and dialysate were sampled at regular intervals during the first hour of dialysis for analysis. RESULTS: Plasma glucose fell below 4.0 mmol/l (72 mg/dl) in 7 of the 18 patients, below 3.5 mmol/l (63 mg/dl) in 3 and below 3.0 mmol/l (54 mg/dl) in one. The lowest recorded value was 2.8 mmol/l (50 mg/dl). The mean glucose loss in the waste dialysate was 9.2 g/h. In none of the group of 7 patients whose plasma glucose fell below 4.0 mmol/l (72 mg/dl) did symptoms of hypoglycemia occur. When 6 patients from this group were subsequently dialyzed with a dialysis fluid containing 5.5 mmol/l (100 mg/dl) glucose, their plasma glucose became stabilized within the fasting reference range. There were no significant hormonal changes during the dialysis or between non-hypoglycemic and hypoglycemic patients. CONCLUSIONS: Patients undergoing hemodialysis may become hypoglycemic and not be aware of it. There is no hormonal imbalance causing the hypoglycemia and the hormonal response to the hypoglycemia is blunted. Patients with an initial plasma glucose of 5.5 mmol/l (100 mg/dl) or less who are hemodialyzed and who do not eat during dialysis may be particularly at risk, especially if they are on insulin or taking glucose-lowering medication. These should be dialyzed with a dialysis fluid containing at least 5.5 mmol/l (100 mg/dl) glucose.

Occult hypoglycemia caused by hemodialysis.

BACKGROUND: Previous studies have ignored hypoglycemia in patients undergoing hemodialysis. The fall in plasma glucose may not have been considered to be clinically relevant because the patients were asymptomatic. The present study was designed to assess the effect of hemodialysis on plasma glucose, insulin, glucagon, cortisol and catecholamines in non diabetic patients. METHODS: 21 non diabetic patients with chronic renal failure were hemodialyzed using a glucose-free dialysis fluid. They did not take any medication prior to dialysis and were asked not to eat during the first hour on hemodialysis. Blood and dialysate fluid was sampled at regular intervals during the first hour of dialysis for analysis. RESULTS: Plasma glucose fell below 4.0 mmol/l (72 mg/dl) in 9 of the 21 patients, below 3.5 mmol/l (63 mg/dl) in 6 and below 3.0 mmol/l (54 mg/dl) in 3. The lowest recorded value was 2.1 mmol/l (38 mg/dl). The mode glucose loss in the waste dialysate fluid was 6 g/h. In the group of 9 patients whose plasma glucose fell below 4.0 mmol/l (72 mg/dl), no symptoms of hypoglycemia were shown but 4 of the 7 patients who felt very hungry and ate were in this group. When 7 patients from this group were subsequently dialysed with a dialysis fluid containing 5.5 mmol/l (100 mg/dl) glucose, their plasma glucose became stabilized within the fasting reference range. There were no significant hormonal changes during the dialysis or between euglycemic and hypoglycemic patients. CONCLUSIONS: Patients undergoing hemodialysis may become hypoglycemic and not be aware of it. There is no hormonal imbalance causing the hypoglycemia and the hormonal response to the hypoglycemia is blunted. Patients with an initial plasma glucose of 4.5 mmol/l (81 mg/dl) or less who are hemodialyzed and who do not eat during dialysis may be particularly at risk. They should be dialysed with a dialysis fluid containing at least 5.5 mmol/l (100 mg/dl) glucose.

Transmural variation of myocardial attenuation measured with a clinical imager.

The objective of this study was to quantify the transmural variation in attenuation for the septal and lateral walls of the heart. Our approach was to utilize a commercially available ultrasonic imaging system to acquire images of excised sections of eight sheep hearts with an orientation similar to that encountered in the apical four-chamber view. The measured values (mean +/-SE) of the slope of attenuation for the transmural regions of the septum are: 1.40 +/-0.11, 0.99 +/-0.09, and 1.85 +/-0.16 (dB/cm/MHz) for the left subendocardial, midmyocardial, and right subendocardial zones, respectively. The analogous data from the lateral wall are: 1.42 +/-0.11, 0.83 +/-0.07, and 1.20 +/- 0.16 (dB/cm/MHz) for the subendocardial, midmyocardial, and subepicardial zones, respectively. These data demonstrate that ultrasonic attenuation associated with the septum and the lateral wall, when imaged in a manner similar to that of the apical four-chamber view, is anisotropic.

Dependence of "apparent" magnitude on the time delay of cyclic variation of myocardial backscatter.

The goal of this study was to determine if the "apparent" magnitude of the cyclic variation, defined as the difference between the values of integrated backscatter at end-diastole and end-systole, was dependent on the corresponding time delay. We measured the cyclic variation in four myocardial segments of the parasternal short-axis view in 23 healthy subjects. The "apparent" magnitude, actual magnitude, and time delay were compared for each segment. Measured time delays were: 2.22+/-0.71 (lateral wall); 1.65+/-0.66 (inferior septum); and approximately 1.0 for the anterior septum and posterior wall. Segments exhibiting large time delays (> 1.0) resulted in a reversal in sign of the "apparent" magnitude of cyclic variation in one instance, and underestimated the true magnitude in both cases. Thus, estimates of the "apparent" magnitude of the cyclic variation are dependent on the associated time delay, whereas a properly defined magnitude is not.

Changing clinicians' laboratory test requesting behaviour: can the poacher turn gamekeeper?

A major reason for the dramatic increase in laboratory costs is the increased number of laboratory investigations, many of which are perceived to be a waste of time and money. In consequence, several strategies, reviewed below, for curtailing clinicians' laboratory usage have been proposed but none has been universally successful. It is clear, however, that a combination of several strategies is likely to be more successful than any single approach. There may be several reasons why no single strategy has been widely adopted in routine practice. We suggest that modern computers have the ability to overcome these obstacles. We propose, therefore, that expert systems at the clinical-laboratory interface utilising a multifactorial approach within a framework of commitment from clinicians and laboratorians offer the best prospect for reducing inappropriate laboratory usage.

Lipaemia: an overrated interference?

Reagent method sheets for analysis of common serum analytes often highlight the possibility of interference from lipaemia but the information given is often brief and may not be instrument-specific. Thus study assesses the degree of interference from lipaemia in a range of common serum analytes on the Bayer Opera (with a serum blank) using a commercial polymer, LipoClear, as a lipid-clearing agent. Serum samples (mean serum triglyceride 6.89 [range 0.58-28.4] mmol/L) are analysed for 14 common chemistry analytes and the results compared before and after treatment with LipoClear. Results showed no significant critical differences in analyte values before and after treatment, except for an expected fall in total protein, phosphate, cholesterol and triglyceride concentrations. Most of the common analytes in use on the Bayer Opera are not subject to interference from lipaemia; however, we recommend that where method sheets indicate interference from lipaemia then this should be quantified for the analyte in question.