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Major pitfalls in Doppler investigations with particular reference to the cerebral vascular system. Part I. Sources of error, resulting pitfalls and measures to prevent errors.

Major pitfalls in Doppler investigations are presented based on 340 evaluated cerebral Doppler examinations in infants. Substantial pitfalls may result from: A. Physics of sound waves and Doppler instruments (errors due to high pass filter cut off, aliasing, rapid image update). B. Quality and adjustment of the Doppler instrument (errors due to low sensitivity, inappropriate adjustment of Doppler controls, inadequate wall filter). C. Examination technique (errors due to an unfavourable angle of incidence or due to transducer-induced pressure: decrease predominantly in diastolic flow velocity-increase in maximum flow velocity in the straight sinus). D. Hemodynamics (errors due to spatial or temporal variations of the flow profile, pulsatility, non-uniform distribution of cerebral blood flow/CBF). E. Cerebral vascular anatomy (errors due to an unfavourable probe position as related to the three-dimensional arrangement of vessels, inadequate separation of closely adjacent vessels). F. Interpretation (flow velocity or Resistance Index/RI is taken to equal CBF, RI is taken to equal peripheral vascular resistance, one artery is taken to represent the cerebral circulation). Pitfalls may be avoided by using adequate means (low wall filter adjustment, high Doppler frequency, critical assessment of velocity spectra) to reduce the likelihood of errors occurring.

Adolescent

Increased left ventricular ejection fraction after a meal: potential source of error in performance of radionuclide angiography.

The effect of a standardized meal on left ventricular (LV) ejection fraction (EF) was determined by equilibrium radionuclide angiography in 16 patients with stable congestive heart failure but without pulmonary or valvular heart disease. LVEF was determined in the fasting state and 15, 30, and 45 minutes after a meal. Patients with moderately depressed fasting LVEF (30 to 50%), Group I, had a mean increase of 6.9 +/- 2.9% (p less than 0.005) in the LVEF at 45 minutes after the meal. Patients with severely depressed fasting LVEF (less than 30%), Group II, had no change after the meal. It is concluded that significant increases in LVEF may occur after meals in patients with moderate but not severe left ventricular dysfunction. Equilibrium radionuclide angiography studies that are not standardized for patients' mealtimes may introduce an important unmeasured variable that will affect the validity of data in serial studies of left ventricular function.

Adult

Cysteine: a potential source of error in amino acid analysis of mercaptoethane sulfonic or hydrochloric acid hydrolysates of proteins and peptides.

Hydrolysis of proteins and peptides with mercaptoethane sulfonic acid is liable to produce overestimation of the proline content owing to the production of ninhydrin-positive material (probably cysteine) which coelutes with proline on many ion-exchange analytical systems. A similar error occurs with HCl hydrolysis (especially in the presence of mercaptoethanol or thioglycollic acid) if care is not taken to oxidize cysteine during reconstitution of the hydrolysate before amino acid analysis.

Amino Acids

Sources of error in estimating radioactivity in protein from cell cultures by liquid scintillation counting.

In this study, several common sample preparation techniques for liquid scintillation counting were critically reviewed. It has been shown that techniques such as NaOH or formic acid solubilization of trichloroacetic acid (TCA)-precipitated proteins led to underestimation of the radioactivity by 20-40%; this loss was not corrected by either internal or external standardization. Hydrolysis of the proteins with 6 N HCl or Pronase significantly increased the recovery of the labeled proteins. Also, 10% of the labeled cell proteins remained on the dish when cells were scraped into buffer; these labelled proteins could be recovered either by in situ hydrolysis with Pronase or by solubilization and scraping in 0.3 N NaOH. These techniques increased the recovered radioactivity by 50-60%, allowing quantitative measurements to be made over a 3-day chase period. A possible mechanism and the implications of this observation were discussed.

Animals