A radio-enzymatic method for the estimation of L-leucine-specific radioactivity

A method is presented for the estimation of L-leucine concentration and radioactivity in biological samples. The sample L-leucine is specifically bound to tRNA and its radioactivity estimated in the presence of either added labelled L-leucine or cold L-leucine (in the same proportion), as well as in the presence of a large excess of cold L-leucine. The latter gave the measurement of non-leucine radioactivity present in the sample. The measurements in the presence and absence of labelled/cold L-leucine allowed the estimation of L-leucine levels and radioactivity by using a simple set of calculations and a standard curve built with known cold L-leucine concentrations in the presence of a fixed known amount of [14C]L-leucine.     

Care pathways: have they a place in ''the new National Health Service''?

Radioactivity after administration of 123I-sodium iodide was measured in breast milk samples obtained from a patient with postpartum thyroiditis. The breast milk was collected over 93 h during the infant's regular feeding times. The radioactivity in the breast milk was calculated with a 123I capsule of the same lot number as the standard source. 123I was excreted exponentially with an effective half-life of 5.5 h; 2.5% of the total radioactivity administered was excreted in the breast milk over the 93 h, 95% of which was excreted within the first 24 h, and 98.2% within 36 h. The first milk sample collected at 7 h after administration of the radiopharmaceutical contained 48.5% of the total radioactivity excreted. We estimated the potential absorption of radioactivity to an infant's thyroid in uninterrupted breast-feeding to be 30.3 mGy. With a 24-hour interruption, the absorbed radioactivity would be 1.25 mGy; with a 36-hour interruption, it would be 0.24 mGy. According to our calculations, breast feeding should be curtailed for 36 h to reduce the infant's exposure to 123I radioactivity. By using a correction factor based on maximum radioactivity from another 123I capsule of the same lot, we were able to ascertain the appropriate protocol for our patient and establish a measurement method that can be applied in similar clinical situations.     

Attempted diagnosis of ventricular mural thrombi in acute mycardial infarction using 125I-labelled fibrinogen

In an attempt to diagnose ventricular mural thrombi complicating acute myocardial infarction (AMI), 80 patients have been given 100 muCi 125I-labelled fibrinogen after admission to a CCU. Precordial radioactivity was recorded for the following 6 days over four sites corresponding to chest leads CR1-CR4. A sustained rise in radioactivity of at least 15% of initial recordings was classed as type A pattern, a minor rise or flattened response as type B pattern and a rapid decrease as type C pattern; 28% showed a type A, 19% a type B and 54% a type C pattern. There was no significant difference between the groups in incidence of pericardial friction rub but when patients with suspected pericarditis (as evidenced by characteristic pains) were added, pericarditis was significantly overrepresented in the type A group. Smaller infarctions (SGOT less than 100 U/1) were significantly more common in patients with a type C decay pattern. No differences were noted between the groups as regards type and site of the infarction. A sustained rise in precordial radioactivity after an AMI may be an indication of mural thrombosis but the influence of other factors secondary to an infarction, e.g. pericarditis, cannot be determined at present.     

111Indium-diethylenetriamine pentaacetic acid cerebrospinal fluid flow studies predict distribution of intrathecally administered chemotherapy and outcome in patients with leptomeningeal metastases

Abnormal CSF flow can impair the distribution of intrathecally administered drugs. We examined the relationship between 111indium-diethylenetriamine pentaacetic acid (111In-DTPA) CSF flow studies and methotrexate levels in ventricular and lumbar CSF and correlated these findings with outcome in patients with leptomeningeal metastases (LM). Seven men and 10 women with LM (10 solid tumors, 6 lymphoma, 1 leukemia) received 12 mg methotrexate and 0.5 mCi 111In-DTPA by intra-Ommaya injection; images were obtained immediately and after 4, 24, and 48 hours. Ventricular and lumbar CSF methotrexate and radioactivity levels were measured 6 hours after injection. Thirteen patients had abnormal CSF flow studies, 9 with multiple sites of obstruction. CSF flow obstruction was observed at ventricular outlets in 13 patients, cerebral convexities in 9 and in the spine in 2. With one exception, all obstructions were explicable by tumor deposits on MRIs. For all patients, ventricular and lumbar methotrexate and radioactivity levels correlated closely. Three patients with a normal CSF flow study are alive at 15+, 7.5+, and 3.9+ months from treatment. Of 12 with abnormal CSF flow studies, 11 are dead a median of 2 months from diagnosis. Two patients had diffusely delayed flow studies and both developed methotrexate leukoencephalopathy. CSF flow studies using 111In-DTPA reliably predict distribution of intrathecal methotrexate. Abnormal flow studies correlate with structural abnormalities, are an unfavorable prognostic factor, and may predict intrathecal chemotherapy toxicity.     

Biosynthesis of camalexin from tryptophan pathway intermediates in cell-suspension cultures of Arabidopsis

Camalexin (3-thiazol-2'-yl-indole) is the principal phytoalexin that accumulates in Arabidopsis after infection by fungi or bacteria. Camalexin accumulation was detectable in Arabidopsis cell-suspension cultures 3 to 5 h after inoculation with Cochliobolus carbonum (Race 1), and then increased rapidly from 7 to 24 h after inoculation. Levels of radioactivity incorporated into camalexin during a 1.5-h pulse labeling with [14C]anthranilate also increased with time after fungal inoculation. The levels of radioactive incorporation into camalexin increased rapidly between 7 and 18 h after inoculation, and then decreased along with camalexin accumulation. Relatively low levels of radioactivity from [14C]anthranilate incorporated into camalexin in the noninoculated controls. Autoradiographic analysis of the accumulation of chloroform-extractable metabolites labeled with [14C]anthranilate revealed a transient increase in the incorporation of radioactivity into indole in fungus-inoculated Arabidopsis cell cultures. The time-course measurement of radioactive incorporation into camalexin during a 1.5-h pulse labeling with [14C]indole was similar to that with [14C]anthranilate. These data suggest that indole destined for camalexin synthesis is produced by a separate enzymatic reaction that does not involve tryptophan synthase.     

Endocardial versus midwall measurement of left ventricular function in mild hypertension: an insight from the Harvest Study

OBJECTIVE: To compare endocardial and midwall measurement of left ventricular fractional shortening in assessing cardiac systolic function in hypertension. SETTING: Seventeen hypertension clinics in northeast Italy. MAIN OUTCOME MEASURES: Left ventricular endocardial fractional shorteningcircumferential stress relationship versus midwall shortening-stress relationship in the subjects divided according to relative wall thickness (RWT) and left ventricular mass indexed by body surface area. PATIENTS: Borderline-to-mild hypertensives [n = 635, aged 33 +/- 0.3 years (mean +/- SEM), office blood pressure 146 +/- 0.4/94 +/- 0.2 mmHg (means +/- SEM)] in the Harvest Study and 50 normotensive controls with similar age and sex distributions. METHODS: Blood pressure was measured by 24 h ambulatory monitoring. Left ventricular dimensional and functional indices were assessed by M-mode echocardiography. RESULTS: In the subjects divided into quintiles of RWT, the left ventricular shortening-stress relationship was increased in a parallel fashion when calculated by endocardial and by midwall measurements for RWT < or = 0.35. Instead, for greater RWT values (> or = 0.37) endocardial measurement constantly gave large values than did midwall measurement. Both the endocardial and the midwall shortening-stress relationships progressively decreased with increasing RWT. However, the endocardial shortening-stress relationship remained greater than normal at any RWT, whereas the midwall shortening-stress relationship was decreased for RWT > or = 0.37. In a multiple-regression analysis RWT was the most potent predictor of the endocardialmidwall shortening difference, left ventricular mass and 24 h systolic blood pressure being the second and third most potent predictors. CONCLUSIONS: We found a parallel increase in indices of cavity emptying and of myocardial contractility in mild hypertensive subjects with normal left ventricular geometry. When the RWT is increased, ejection phase indices may be normal in the presence of decreased myocardial contractility.     

Assessment of left ventricular volume by contrast echocardiography

Two-dimensional echocardiography for measuring left ventricular volumes usually gives volumes that are smaller than those determined with left ventriculography. This is due to less optimal image quality since manual tracing of endocardial borders requires still frames. Intravenous injection of echocontrast agent (Albunex) improve endocardial border recognition and therefore left ventricular volume measurements become more accurate. It is reported that contrast echocardiography significantly improves the correlation of echocardiographic left ventricular volume measurement with that of left ventriculography. From this points of view, contrast echocardiography is useful for the determination of left ventricular volumes in clinical settings.     

Assessment of 123I-beta-methyl iodophenyl pentadecanoic acid (BMIPP) myocardial scintigraphy in patients of chronic right ventricular overload--fatty acid metabolism in right ventricular myocardium

An investigation on the right ventricular pressure level and the abnormalities in the fatty acid metabolism of myocardium was made using 123I-beta-methyl-iodophenyl pentadecanoic acid (BMIPP) myocardial SPECT in patients with chronic right ventricular overloading. Twenty patients who presented with right ventricular systolic pressure (RVSP) of 35 mmHg or more were used as the subjects. Dual myocardial SPECT with 201TlCl (Tl) and BMIPP was carried out for the subjects and RVc/LVc, a ratio of radioactivity count incorporated in the right ventricular free wall to the left one was determined for Tl and BMIPP. And the correlations between RVc/LVc and RVSP, and RVc/LVc and RVSP/LVSP were examined. The subjects were classified into 3 groups based on the RVSP levels and the count ratio, BMIPP/Tl was compared among the three groups. With respect of Tl uptake, there were significant, positive correlations between RVc/LVc and RVSP (correlation coefficient r = 0.51, p < 0.05) and between RVc/LVc and RVSP/LVSP (correlation coefficient r = 0.59, p < 0.01). On the other hand, no significant correlation was found between them with respect of the uptake of BMIPP. The BMIPP/Tl ratio in the group with higher than 80 mmHg of RVSP was 0.82 +/- 0.06, which was significantly lower than the ratio's for two groups of less than 80 mmHg; 0.91 +/- 0.07 and 0.98 +/- 0.04 in the group with 35-49 and 50-79 mmHg of RVSP, respectively. These results show that when compared with BMIPP, Tl is superior for the estimation of right ventricular pressure. For the patients with right ventricular overloading, it was suggested that when RVSP reaches 80 mmHg or more, there appear some disorders in the fatty acid metabolism in the right ventricular myocardium.     

A simple technique for measuring relative renal blood flow

To determine whether externally monitored early renal uptake of 131I-hippurate is proportional to renal blood flow, the renal uptake of 131I-hippurate at 1--2 min after injection was compared with the renal accumulation of radioactive carbonized microspheres in dogs. A renal artery catheter equipped with a balloon was used to decrease renal blood flow unilaterally. One minute after the intravenous injection of 100 muCi of 131I-hippurate, about 1 muCi of either 85Sr- or 98Nb-labeled carbon microspheres was injected into the left ventricle. Radioactivity was measured over both kidneys. The total radioactivity within each kidney region of interest was corrected for background and integrated over the 1--2 min after injection. Thirteen measurements of relative renal blood flow were made for seven dogs. The dogs were then killed and both kidneys were excised and counted for the radioactivity of the microspheres. The 1--2-min relative renal uptake of 131I-hippurate correlated well with relative microsphere uptake, suggesting that relative renal blood flow can be simply determined from the external measurement of renal uptake of 131I-hippurate.     

Quantitation of phosphatidyl N-methyl-and N,N-dimethylaminoethanol in rat liver

The contents of phosphatidyl N-methyl- and N,N-dimethylaminoethanol were determined in the liver of rats injected with (Me-C14) methionine. Total phospholipids were extracted from aliquots of the liver and fractionated by two-dimensional thin layer chromatography after addition of carrier phosphatidyl-N-methyl- and N,N-dimethylaminoethanol. The radioactivity present in the two phosphatide spots was determined and used to calculate total disintegrations per min/100 g body wt. The remainder of the livers was pooled, and total phospholipids were isolated and subjected to acid hydrolysis. N-methyl- and N,N-dimethylaminoethanol were purified by thin layer chromatography, and their specific activity was determined after quantitation by gas liquid chromatography and radioactivity measurement. The liver contents of phosphatidyl N-methyl- and N,N-dimethylaminoethanol were determined by dividing disintegrations per min/100 g body wt by the specific activity of N-methyl- or N,N-dimethylaminoethanol.     

Reproducibility of color Doppler flow quantification of aortic regurgitation

The preferred method for quantification of aortic regurgitation severity with color Doppler echocardiography is the assessment of the ratio of jet diameter to left ventricular outflow tract diameter and jet area to left ventricular outflow tract area. However, the reproducibility of these measurements is not known and may limit its clinical application. This study was performed to identify sources of variability and reproducibility of the echocardiographic data. We examined 62 color Doppler echocardiographic examinations of patients showing isolated aortic regurgitation after human tissue valve implantation. The mean differences with standard deviations between paired measurements were calculated. The interobserver, intraobserver, and interframe variability showed a close agreement for the jet diameter and left ventricular outflow tract diameter measurements. The agreement for jet area and left ventricular outflow tract area measurements showed a small bias, but a large variance. The reproducibility of jet-left ventricular outflow tract diameter is better than the jet-left ventricular outflow tract area measurement and is more accurate to assess the severity of aortic regurgitation from color Doppler images.     

Left ventricular function during respiratory failure

Indirect measures of left ventricular function were studied in seven patients with respiratory failure secondary to chronic obstructive pulmonary disease to determine if there were a relationship between left ventricular function and treatment of the pulmonary disease. All patients were studied during acute episodes while in respiratory failure having arterial Pco2 (Paco2) values greater than 49 torr with no clinical evidence of left ventricular failure. Indirect methods to evaluate left ventricular function included the use of the Swan-Ganz catheter for pulmonary capillary wedge pressure measurement, systolic time intervals, and cardiac output. There was improvement in left ventricular function with treatment of the respiratory failure manifested by decreases in the wedge pressure and pre-ejection period/left ventricular ejection time ratio, and an increase in the dp/dt/pulmonary capillary wedge pressure with treatment of the chronic obstructive pulmonary disease. The improvement in left ventricular function suggests that there is a depression of left ventricular function in respiratory failure. The depressed function improved with therapy of the lung disease without additional medication directed at cardiac function.     

Hemodynamic basis of mitral E transmission in the left ventricular cavity and its relation to the left ventricular relaxation process

Mitral E-wave transit time to the left ventricular outflow tract was measured as an E-Er interval in 30 subjects undergoing cardiac catheterization. The E-Er interval (range 30 to 190 ms) correlated with left ventricular peak negative dP/dt (r = -0.62, p = 0.0003) and tau (r = 0.74, p <0.0001) but not with left ventricular minimum, pre-A-wave, or end-diastolic pressures. We conclude that the E-Er interval is an easily obtainable Doppler measurement that reflects the left ventricular relaxation process.     

Relationship between left ventricular mass and serum cholesterol level in the untreated hypertensive

OBJECTIVE: To study the relationship between serum cholesterol level and left ventricular mass for a population of untreated hypertensive patients. DESIGN: A cross-sectional study. PATIENTS: We studied 273 untreated hypertensive patients without associated diseases consecutively referred for evaluation of blood pressure. All patients underwent M-mode echocardiographic assessment of left ventricular mass, office blood pressure measurement and 24 h ambulatory blood pressure monitoring. Fasting plasma glucose and total cholesterol levels were measured on the same day. RESULTS: We found a weak but significant correlation (r = 0.20-0.26, P < 0.01) between serum cholesterol level and left ventricular wall thickness or left ventricular mass irrespective of the mode of indexation used (height, height2.7 and body surface area). In multivariate analysis this relation remained significant after introduction of sex, age, weight, height, blood pressure and blood glucose level. When data for men and women were analysed separately the relationship between left ventricular mass and cholesterol remained significant for men only. CONCLUSION: There is a significant and independent positive relationship between serum cholesterol level and left ventricular mass that could contribute to the prognostic value of left ventricular hypertrophy.     

Ferriprotoporphyrin catalysed decomposition of artemether: analytical and pharmacological implications

The ability of the products of erythrocytic haemolysis and ferriprotoporphyrin (FP-IX)-containing substances of facilitate the decomposition of the antimalarial drug artemether has been evaluated in vitro. The products of haemolysis accelerate the degradation of artemether to unidentified compounds which are undetectable by currently available HPLC methods. This decomposition is temperature dependent and occurs after relatively brief artemether (ARM)-catalyst contact. Using radiolabelled [16-14C]ARM, we have demonstrated that the extractability of total radioactivity into the organic phase diminishes with increasing FP-IX concentration with an appreciable reduction in the organic extractability as ARM in the presence of FP-IX and associated increase in water solubility of radioactivity when the concentration of FP-IX increases from 0 to 7.7 mM. This effect appears to be temperature dependent for incubations with haematin. Characterisation of the organically extractable radioactivity from incubations of [16-14C]ARM with FP-IX has shown a loss of ARM and the formation of two radioactive products which occurs in proportion to the concentration of FP-IX. We believe these findings are of particular relevance to the determination of plasma concentrations of ARM and dihydroartemisinin (DHA) in malaria patients where extensive haemolysis and the presence of breakdown products of haemoglobin may contribute to a reduction in the circulating concentration of these substances. In these circumstances, measurement of ARM and DHA by conventional methods may be meaningless.     

Flow in pulmonary veins

Pulmonary venous flow can be evaluated by means of pulsed Doppler echocardiography during a transthoracic or transesophageal echocardiography examination. In most cases a three-peak spectral curve with highest positive wave during ventricular systole, a lower positive wave during ventricular diastole and a small negative wave during the left atrial contraction are recorded. Multiple factors affect the systolic and diastolic components of pulmonary venous flow. Pulmonary venous flow measurement may prove to be an important tool in routine judgment of diastolic left ventricular and atrial function. With regard to clinical importance, the pulmonary venous flow is a useful parameter for left atrial pressure quantification and for differential diagnosis of restrictive cardiomyopathy and constrictive pericarditis.     

Real-time continuous measurement of right ventricular volume using a conductance catheter

The authors propose using a multi-electrode conductance catheter to measure continuous right ventricular volume. True ventricular volume measurements are affected by four main sources of error. 1) field non-uniformity, 2) catheter curvature, 3) blood conductivity changes, and 4) leakage of current through surrounding tissues. Three-dimensional finite-element models were developed to investigate the effects of these sources of error and to devise schemes for correcting them. The models include an axisymmetric cylindrical model, a rectangular block model, and a heart model with left and right ventricular chambers. The heart model is built from conical primitives, with major dimensions derived from the literature. Finite-element simulations showed that volume measurements were underestimated due to field nonuniformity to as much as 1/25th actual volume in segments near the exciting electrodes. The extent of underestimation in a segment decreased with increasing distance of the segment from the exciting electrodes and increased for larger segmental volumes. Catheter curvature overestimated measured volume by as much as 4.5 times when the curvature was increased from 0.0 to 1.25 (from a straight catheter to a very curved one). The leakage of current through surrounding tissues overestimated volume by nearly 30%. The sensitivity of volume measurement to blood resistivity changes was found to be very high, at 70%. Correction factors established with the computer models compensate for field nonuniformity. Mathematical mapping of the curved catheter onto a fictitious straight catheter corrects for the catheter curvature error. Correction for both nonuniform field and catheter curvature allowed measurement of total ventricular volume with an error of 7%. Leakage current is determined by using different frequencies to build the catheter electric field and to separate tissue and blood resistance paths. Using this scheme, the percentage overestimation in volume measurement due to leakage could be determined with an accuracy of 85%. The proposed correction scheme for blood conductivity changes involves the in-vivo measurement of blood conductivity with the catheter itself. It was found that blood conductivity could be determined with insignificant error (< 0.5%) so long as the blood volume around the exciting electrodes had a radius of more than the electrode spacing.     

Noninvasive quantification of regional myocardial metabolic rate for oxygen by use of 15O2 inhalation and positron emission tomography. Theory, error analysis, and application in humans

BACKGROUND: A method has been developed to measure the regional myocardial metabolic rate of oxygen consumption (rMMRO2) and oxygen extraction fraction (rOEF) quantitatively and noninvasively in humans by use of 15O2 inhalation and positron emission tomography. This article describes the theory, an error analysis of the technique, and procedures of the method used in a human feasibility study. METHODS AND RESULTS: Inhaled 15O2 is transported to peripheral tissues, where it is converted to 15O-labeled water of metabolism, which exchanges with the relatively large extravascular tissue space. Quantification of this buildup of radioactivity allows the calculation of rMMRO2 and rOEF. However, a correction for the spillover of the pulmonary gas radioactivity signal into myocardial regions is required and has been made by use of a gas volume distribution estimated from the transmission scan. This was validated by comparative measurements using the inert gas [11C]CH4 in four greyhounds. Spillover of the cardiac chamber radioactivity has been corrected for with an inhaled [13O]CO (blood volume) scan. The underestimation of myocardial radioactivity due to wall motion and thickness has been corrected for by use of values of tissue fraction obtained from the flow measurement [15OKCO2 scan). Values of rOEF were similar (within 4%) whether obtained from gas volume measurements determined from the transmission or [11C]CH4 scan data. 15O2 scan information from six healthy volunteers showed a clear distribution of myocardial radioactivity after the vascular and pulmonary gas 15O background was subtracted. Subsequent compartmental analysis resulted in values for rOEF and rMMRO2 of 0.60 +/- 0.11 and 0.10 +/- 0.03 mL.min-1.g-1 in the human myocardium at rest. CONCLUSIONS: The results of this study are in good agreement with established values. This is the first known approach to allow the direct quantitative determination of rOEF and oxygen metabolism to be made noninvasively on a regional basis.     

Calibration methods for measuring splenic sequestration by external scanning

Conventional methods for measuring splenic sequestration of labeled cells rely on stationary probe counting over the liver and spleen. A quantitative spleen-scanning method is proposed as a means of performing a more accurate diagnosis of the degree of splenic sequestration. Improved methods for calibrating a rectilinear scanner employing "constant resolutions" collimators, designed for the in vivo measurement, are reported, Clinical evaluation of this method was performed by scanning several patients scheduled for splenectomy. Good correlation between in vivo and in vitro measurements of splenic radioactivity for 29 patients was obtained. A least-squares fit to the data yielded a coefficient of determination r2=0.93.     

Assessment of regional myocardial uptake and metabolism of omega-(p-123I-phenyl) pentadecanoic acid with serial single-photon emission tomography

The utility of myocardial imaging and assessment of regional myocardial metabolism of omega-(123I-paraphenyl-)pentadecanoic acid (I-PPA) by means of serial single-photon tomography is demonstrated in animal experiments. High quality cross sectional images of dog hearts with clear delineation of left ventricular walls are obtained. Myocardial infarcts are visualized as areas of deficient radioactivity uptake. I-PPA elimination from non-infarcted myocardial regions is significantly (p less than 0.001) prolonged when compared with unaffected controls. Hence, not only localized absence of uptake of free fatty acid by infarcted myocardium can be demonstrated with serial single-photon tomography but also general impairment of cardiac FFA-metabolism.