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Ultrasound-guided optical tomographic imaging of malignant and benign breast lesions: initial clinical results of 19 cases.

The diagnosis of solid benign and malignant tumors presents a unique challenge to all noninvasive imaging modalities. Ultrasound is used in conjunction with mammography to differentiate simple cysts from solid lesions. However, the overlapping appearances of benign and malignant lesions make ultrasound less useful in differentiating solid lesions, resulting in a large number of benign biopsies. Optical tomography using near-infrared diffused light has great potential for imaging functional parameters of 1) tumor hemoglobin concentration, 2) oxygen saturation, and 3) metabolism, as well as other tumor distinguishing characteristics. These parameters can differentiate benign from malignant lesions. However, optical tomography, when used alone, suffers from low spatial resolution and target localization uncertainty due to intensive light scattering. Our aim is to combine diffused light imaging with ultrasound in a novel way for the detection and diagnosis of solid lesions. Initial findings of two early-stage invasive carcinomas, one combined fibroadenoma and fibrocystic change with scattered foci of lobular neoplasia/lobular carcinoma in situ, and 16 benign lesions are reported in this paper. The invasive cancer cases reveal about two-fold greater total hemoglobin concentration (mean 119 micromol) than benign cases (mean 67 micromol), and suggest that the discrimination of benign and malignant breast lesions might be enhanced by this type of achievable optical quantification with ultrasound localization. Furthermore, the small invasive cancers are well localized and have wavelength-dependent appearance in optical absorption maps, whereas the benign lesions appear diffused and relatively wavelength-independent.

Adult↗

Continuous-flow isotope analysis of the deuterium/hydrogen ratio in atmospheric hydrogen.

A convenient method is described for analyzing the deuterium/hydrogen (D/H) ratio of atmospheric molecular hydrogen (H(2)) based on mass spectrometric isotope-ratio monitoring. The method requires small amounts of air ( approximately 300 mL STP), is operated on-line, and comprises four steps: (1). the condensation of the air matrix at approximately 40 K; (2). the collection of the non-condensed components of the air sample (H(2), Ne, He, and traces of N(2)) in a 5 A molecular sieves pre-concentration trap at approximately 63 K; (3). gas chromatographic purification of H(2) in a flow of He; and (4) quantification of the D/H ratio in an isotope-ratio mass spectrometer. The precision of the determination of the D/H ratio is better than 2 per thousand, which is comparable to, or better than, that obtained by conventional duel-inlet off-line analysis. There are, however, discrepancies relative to the D/H ratios determined by conventional duel-inlet analysis. This is due to differences in peak shape between reference and sample air, depending on the amount of H(2) injected. Consequently, calibration runs are required. After the calibration of the system, we obtained an accuracy of 1.5 per thousand, so that the accumulated uncertainty is estimated to be less than 4 per thousand. The method also allows determination of the H(2) concentration, with an uncertainty estimated to be 2%.

Atmosphere↗

Can lithium-heparin plasma be used for protein electrophoresis and paraprotein identification?

INTRODUCTION: Lithium-heparin plasma is the most commonly used sample type in many hospitals, but it has been suggested that it is not suitable for protein electrophoresis due to the presence of fibrinogen, which can potentially mask a paraprotein band or be misconstrued as one. Here we aimed to demonstrate that lithium-heparin plasma samples could be used for protein electrophoresis and paraprotein typing without or with ethanol treatment to remove the fibrinogen. METHOD: A lithium-heparin sample from a patient with IgGlambda, IgGkappa, IgAlambda and IgAkappa myeloma, a non-specific polyclonal increase and a serum control were treated with ethanol prior to protein electrophoresis. Immunofixation electrophoresis was undertaken to investigate the effect of ethanol treatment on immunoglobulin and light chains. Nephelometry was undertaken to investigate whether ethanol treatment affected the quantification of IgG levels. Densitometric evaluation of proteins after electrophoresis was used to study whether ethanol treatment affected other serum proteins. An audit was also undertaken to ascertain the magnitude of the potential interference from the fibrinogen band in heparinized samples. RESULTS AND CONCLUSIONS: Ethanol treatment significantly but incompletely removed the fibrinogen in lithium-heparin plasma samples and did not affect the integrity of any of the proteins investigated. Even without ethanol treatment, lithium-heparin plasma can be used for protein electrophoresis and paraprotein identification as the instances of interference between fibrinogen and paraproteins was low (2.3%). In rare cases where there is uncertainty or ambiguity, immuno-fixation electrophoresis is recommended. This report has implications in terms of reducing costs and turn-around time as it prevents the need for requesting another serum sample from patients. This may be one step towards a universal sample for all tests.

Electrophoresis, Agar Gel↗

Human responses to propionic acid. I. Quantification of within- and between-participant variation in perception by normosmics and anosmics.

The objective of this study was to fully characterize normosmic perception of stimuli expected to cause widely varying degrees of olfactory and nasal trigeminal stimulation and to directly evaluate the possible role of olfactory nerve stimulation in nasal irritation sensitivity. During each of four identical test sessions, four anosmic and 31 normosmic participants were presented with a range of concentrations extending from peri-threshold for normosmics to supra-threshold for anosmics. For each session, odor (O) and nasal irritation (NI) sensitivities were summarized in terms of the concentrations required to produce four sensation levels ('iso-response' concentrations). Within-participant variation in these iso-response concentrations was < 10-fold for 95% of normosmics, for both O and NI. For O but not NI, these apparent fluctuations in sensitivity were largely accounted for by the uncertainty surrounding the iso-response concentrations calculated for each session. Anosmics exhibited minimal within- and between-participant variation in NI and required, for all but the highest perceptual level, a higher concentration than almost all normosmics. Between-participant variation, expressed in terms of 90% confidence interval widths, was approximately 0.5 log units for both O and NI for the highest perceptual level, but increased to approximately 0.8 and 1.8 log units, respectively, for the lowest (peri-threshold) level. Our findings suggest that: (i) most apparent variation over time in O sensitivity is actually a reflection of the uncertainty surrounding estimates of sensitivity obtained for each session; (ii) within- and between-participant variation in O sensitivity is far less than is commonly reported; and (iii) low to moderate levels of NI in normosmics are the result of relatively weak trigeminal stimulation combined with much greater olfactory activation.

Adolescent↗

Development and validation of a liquid chromatographic/tandem mass spectrometric method for the determination of sertraline in human plasma.

A sensitive and rapid liquid chromatographic/tandem mass spectrometric method was developed and validated for the determination of sertraline in human plasma. The analyte and internal standard (IS, diphenhydramine) were extracted with 3 mL of diethyl ether/dichloromethane (2:1, v/v) from 0.25 mL plasma, then separated on a Zorbax Eclipse XDB C18 column using methanol/water/formic acid (75:25:0.1, v/v/v) as the mobile phase. The triple quadrupole mass spectrometry was applied via an atmospheric pressure chemical ionization (APCI) source for detection. The fragmentation pattern of the protonated sertraline was elucidated with the aid of product mass spectra of isotopologous peaks. Quantification was performed using selected reaction monitoring of the transitions of m/z 306 --> 159 for sertraline and m/z 256 --> 167 for the IS. The method was linear over the concentration range of 0.10-100 ng/mL. The intra-day and inter-day precisions, expressed by relative standard deviation, were both less than 6.7%. Assay accuracies were within +/-6.9% as terms of relative error. The lower limit of quantification (LLOQ) was identifiable and reproducible at 0.10 ng/mL with a precision of 8.3% and an accuracy of 9.6%. The validated method has been successfully applied for the pharmacokinetic study and bioequivalence evaluation of sertraline in 18 healthy volunteers after a single oral administration of 50 mg sertraline hydrochloride tablets.

Chromatography, Liquid↗

A routine method for the determination of the TVOC content in wallcoverings using headspace gas-chromatography.

A method for the fast routine analysis of the total content of volatile organic compounds in wallcoverings and paper products was developed, using headspace gas-chromatography for quantification. 57 wallcoverings of different types were investigated. Typical components were toluene, methyl-ethyl ketone, methyl-iso-butyl ketone, n-butyl acetate and iso-butyl acetate, all compounds being used as industrial solvents. The TVOC concentrations are calculated in toluene-carbon equivalents and ranged from 0.31 microg/g to 1789 microg/g with an average value of 123.22 microg/g and a median of 20.37 microg/g. To obtain an estimation of VOC-concentrations in indoor air, 10 selected wallcoverings were also analyzed in a 1 m(3) climate test chamber. A correlation between headspace data and chamber concentrations could not be observed, which might be a result of increased analytical uncertainties at low emission rates under chamber conditions.

Journal Article↗

Analysis of health effects resulting from population exposures to acid precipitation precursors.

Types of available studies relevant to the quantification of air pollution health effects and their principal limitations are discussed. Assessments are provided based on review and re-analysis of previously reported data bases, synthesis of published findings, and original analysis of health data sets using new methods or recent size-specific particle mass measurements. Interim results from ongoing research activities on airborne particle health effects are presented. It is shown that preliminary results obtained from cross-sectional and time-series mortality studies appear to be consistent, indicating that particulate air pollution, even at current levels, could be of concern for public health. Throughout the paper, methodological deficiencies and remaining gaps in knowledge are identified. In particular, uncertainties associated with the reported exposure-response coefficients are assessed. Finally, by characterizing the limitations of analysis we propose various recommendations for future studies and research that will serve to further define the nature, magnitude, and uncertainties of air pollution health risks.

Acids↗

Cosmogenic and nucleogenic isotopic changes in Mars: their rates and implications to the evolutionary history of Martian surface.

We present calculations of rates of production of several nuclides in the Martian atmosphere and in the regolith due to nuclear interactions of cosmic ray and radiogenic particles and consider their implications to the evolutionary history of Mars. Nuclides selected are those which, considering their chemical properties, may be useful as tracers for delineating the past histories of the Martian atmosphere and regolith. Calculations are presented for different assumed atmospheric pressures. The regolith production rates for the present thin Martian atmosphere (approximately 20 g cm-2) are expected to be fairly robust because they are based primarily on observed cosmogenic effects in the Moon, for which semiempirical estimates of nuclide production rates have been provided earlier by Reedy (1981). Uncertainties which arise in the calculations of nuclide production rates for an earlier hypothetical Martian atmosphere of approximately 300-500 g cm-2 thickness are discussed. Compared to cosmic ray production rates, the nucleogenic production rates are smaller by several orders of magnitude. However, the nucleogenic production extends to much deeper levels, whereas the cosmogenic production is essentially confined to the top 750-1000 g cm-2 depth. Important examples of nucleogenic production are discussed. Isotopes of neon and argon appear to be very promising for delineating relative magnitudes of a number of planetary processes related to the temporal changes in the thickness of the atmosphere, as well as their release from the regolith. However, quantification of the processes would require higher-precision isotopic data for the atmosphere and also direct measurements of isotopic ratios in the Martian regolith, along with supplementary information on changes in the isotopic compositions of hydrogen, carbon, and nitrogen, which are affected by a variety of mechanisms of escape of gases from the atmosphere. Cosmogenic effects are minimal in these cases. We show that although we can at present draw but limited inferences, the planet Mars presents a unique opportunity to use cosmogenic nuclides as tools to delineate the evolutionary history of the planet as a whole, as well as its regolith and the atmosphere. This arises because of two factors: minimal degassing of the planet, and a fairly intense chemical weathering history of the upper surface. Consequently, an appreciable fraction of some of the isotopes of volatile elements is contributed by nuclear reactions.

Argon↗

Discontinuation of prophylaxis against Mycobacterium avium complex disease in HIV-infected patients who have a response to antiretroviral therapy. Terry Beirn Community Programs for Clinical Research on AIDS.

BACKGROUND: Several agents are effective in preventing Mycobacterium avium complex disease in patients with advanced human immunodeficiency virus (HIV) infection. However, there is uncertainty about whether prophylaxis should be continued in patients whose CD4+ cell counts have increased substantially with antiviral therapy. METHODS: We conducted a multicenter, double-blind, randomized trial of treatment with azithromycin (1200 mg weekly) as compared with placebo in HIV-infected patients whose CD4+ cell counts had increased from less than 50 to more than 100 per cubic millimeter in response to antiretroviral therapy. The primary end point was M. avium complex disease or bacterial pneumonia. RESULTS: A total of 520 patients entered the study; the median CD4+ cell count at entry was 230 per cubic millimeter. In 48 percent of the patients, the HIV RNA value was below the level of quantification. The median prior nadir CD4+ cell count was 23 per cubic millimeter, and 65 percent of the patients had had an acquired immunodeficiency syndrome-defining illness. During follow-up over a median period of 12 months, there were no episodes of confirmed M. avium complex disease in either group (95 percent confidence interval for the rate of disease in each group, 0 to 1.5 episodes per 100 person-years). Three patients in the azithromycin group (1.2 percent) and five in the placebo group (1.9 percent) had bacterial pneumonia (relative risk in the azithromycin group, 0.60; 95 percent confidence interval, 0.14 to 2.50; P=0.48). Neither the rate of progression of HIV disease nor the mortality rate differed significantly between the two groups. Adverse effects led to discontinuation of the study drug in 19 patients assigned to receive azithromycin (7.4 percent) and in 3 assigned to receive placebo (1.1 percent; relative risk, 6.6; P=0.002). CONCLUSIONS: Azithromycin prophylaxis can safely be withheld in HIV-infected patients whose CD4+ cell counts have increased to more than 100 cells per cubic millimeter in response to antiretroviral therapy.

AIDS-Related Opportunistic Infections↗

Sequential expression of transforming growth factor-beta1 by T-cells, macrophages, and microglia in rat spinal cord during autoimmune inflammation.

Transforming growth factor-beta1 (TGF-beta1) is crucially involved in regulating inflammatory events during experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis. Despite accumulating evidence for local expression of TGF-beta1 in the inflamed nervous system, uncertainty remains regarding its cellular source. We have investigated the temporospatial distribution of TGF-beta1 gene expression in rat spinal cord during EAE. In actively induced EAE, in situ hybridization revealed strong expression of TGF-beta1 in meningeal and perivascular mononuclear infiltrates at onset of the disease, continued expression in perivascular infiltrates and scattered mononuclear cells at maximal disease severity, and expression in scattered parenchymal cells during recovery. Double labeling studies revealed subpopulations of infiltrating T-cells to be the major source of TGF-beta1 early in the disease, followed by macrophages at peak severity and microglial cells during the recovery phase of EAE. Astrocytes and neurons did not express TGF-beta1. Quantification of mRNA by Northern blot analysis revealed that cellular expression of TGF-beta1 by T-cells, macrophages, and microglia sums up to a long-lasting elevation of TGF-beta1 mRNA extending well into the recovery phase. Our data indicate cellular diversity and suggest functional diversity of TGF-beta1 gene expression during EAE. While TGF-beta1 expressed early in the disease by T-cells may contribute to inflammatory lesion development, microglial cells may potentially contribute to recovery by expressing immunosuppressive TGF-beta1 during remission.

Adoptive Transfer↗

Determination of the level of benzo[a]pyrene in fatty foods and food supplements.

A routine method was developed for the quantification of benzo[a]pyrene (BaP) in edible oils and food supplements. BaP is often taken as an indicator of the presence of polycyclic aromatic hydrocarbons. The method consists of on-line liquid chromatography clean-up followed by injection to an HPLC system with fluorescence detection. The method has good performance characteristics and gave good results in proficiency tests. From 2002 to 2004, about 1350 samples of oils and food supplements were analysed using this method to test the level of BaP. About 20% of the edible oils contained more than 1.2 microg kg-1 BaP (which is the limit applied by the Dutch Food and Consumer Product Safety Authority until 1 April 2005, and includes measurement uncertainty). In the case of food supplements, more then 30% contained too high levels of BaP, ranging from 1.2 to 135 microg kg-1.

Benzo(a)pyrene↗

Trace determination of anthracyclines in urine: a new high-performance liquid chromatography/tandem mass spectrometry method for assessing exposure of hospital personnel.

Health-care workers handling antineoplastic agents may be exposed to extremely low doses of these drugs. Very sensitive and specific analytical methods are therefore needed for biological monitoring. The aim of this study was to develop and validate a method for trace level determination of doxorubicin, epirubicin, daunorubicin and idarubicin in human urine, using epi-daunorubicin as an internal standard. Solid-phase extraction (SPE) was used for sample preparation. Urine samples were loaded onto Bond Elut C18 cartridges. The analytes were eluted in methylene chloride/2-propanol (1:1, v/v) and then evaporated to dryness. The residue was reconstituted with the mobile phase prior to high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) analysis. Quantitation of each analyte was performed using the multiple reaction monitoring (MRM) method. The urine assay was linear over the range 0.1-2.0 microg/L, with a lower limit of quantification (LLOQ) of 0.10 microg/L for doxorubicin and epirubicin, and 0.03 microg/L for daunorubicin and idarubicin. The respective limits of detection (LODs) were 0.04 and 0.01 microg/L. The precision and accuracy of the assay were determined on three different days. The within-series precision was found to be always less than 13.9% for all the analytes. The overall precision expressed as relative standard deviation (RSD) was always less than 10.6%. The recovery of anthracyclines was assessed at two concentrations of the range tested (0.1 and 2.0 microg/L) and it ranged from 87.7% (daunorubicin) to 102.0% (doxorubicin) and from 79.1% (daunorubicin) to 90.7% (idarubicin) for the lower and the higher level, respectively, with a RSD always less than 9.1%. The uncertainty of the present assay was also evaluated and the combined uncertainty was always less than 20% over all the days of the validation study. This is the first method that makes use of LC/MS/MS for the biological monitoring of occupational exposure to anthracyclines.

Anthracyclines↗

Correlations between levels of mite and cat allergens in settled and airborne dust.

Thirty homes in Rochester, Minn., 20 of which housed cats, were studied to compare cat and mite allergen concentrations in airborne and settled dust. With Air Sentinels in the bedroom and living room for airborne collections, and a Sample Vac for collections from living room carpet and bedroom mattress, immunochemical quantifications of each were made with various radiometric assays with polyclonal and monoclonal antibodies. The most significant correlations were found between airborne mite in the bedroom and living room (p less than 0.001) and airborne mite in the bedroom and dust from the bedroom mattress (p less than 0.001). Most houses had specific epitopes of both Dermatophagoides pteronyssinus and D. farinae, but D. farinae was present in greater amounts. In seven houses we found greater than 10 micrograms of group I mite allergen per gram of settled dust, indicating that some houses in Minnesota have concentrations of mite allergens high enough to cause allergic disease, even in the winter. Clinical interpretation of these data on air levels is hampered by uncertainty as to whether symptoms are more closely related to average steady-state exposure, which we measured, or to brief heavy concentrations. Further epidemiologic studies are needed to define the concentration of mite and cat allergens in the air that are risk factors for allergic disease. The concentration of cat allergen correlated with mite allergen in the air, but not in settled dust, presumably reflecting the fact that both are influenced by ventilation rate.

Air↗

Apoptosis: measurement and technical issues.

In this short review article we discuss the many uncertainties with the estimation of the relevance of data on the rate of apoptotic cell death in the myocardium. 1. The time course and duration of the apoptotic cascade is still almost completely unknown, in cardiac tissue as well as in myocytes in culture. 2. The mode of removal of dead cells from their connections to neigbouring myocytes at the intercalated disc is still unidentified. 3. At what step in the apoptotic cascade is this process still reversible? 4. Numerous methods exist for the identification of apoptotic cells. However, their sensitivity and specificity vary and issues such as different dwell times of cells with DNA fragmentation further complicate the reliability of apoptotic rates obtained. 5. Statistical principles should be applied for the quantification of apoptotic cells. Only after clarification of these technical issues an unequivocal answer to the problem of apoptotic myocyte loss in different cardiac pathologies can be expected.

Animals↗

Segmented 5 versus nonsegmented flow quantitation: comparison of portal vein flow measurements.

RATIONALE AND OBJECTIVES: Times of acquisition, mean velocities, mean flows, and their respective standard deviations provided by segmented and nonsegmented flow quantifications (FQ) were compared in the main portal veins of ten healthy adults. METHODS: The segmented FQ performed five phase-encoding lines per segment. The authors successively applied the two electrocardiogram-triggered techniques in the same slice perpendicular to the portal flow direction. Their measurements were compared in particular by means of the statistical analysis proposed by Bland and Altman. RESULTS: The segmented 5 FQ reduced the acquisition time by two compared with the nonsegmented FQ. The mean velocity and mean flow values of the two techniques were not significantly different. The standard deviations were similar. CONCLUSIONS: The segmented 5 FQ showed its major advantage in the portal vein: the acquisition time was reduced without any loss of accuracy or any uncertainty enhancement.

Adult↗

Quantification of dose perturbation by plaque in vascular brachytherapy.

BACKGROUND: Dose prescription and reporting in vascular brachytherapy (VBT) is based on the assumption that the vessel wall is water equivalent, which does not consider a possible dose perturbation by plaque. As the extent of this perturbation is unknown, we aimed to quantify dose attenuation by atherosclerotic plaque for beta- and gamma-radiation. MATERIAL AND METHODS: The dose delivered from Strontium-90/Yttrium-90 ((90)Sr/Y) and Iridium-192 ((192)Ir) sources with and without human peripheral arteries ((90)Sr/Y: n = 38, (192)Ir: n = 7) surrounding the respective delivery catheter was determined with radiochromic films. Plaque and vessel wall thickness were measured using light microscopy. From the ratio-attenuated doseunattenuated dose (dose perturbation factor: DPF) we determined averaged attenuation coefficients for atherosclerotic plaque (micro(P)) and the residual part of the vessel wall (micro(W)) by regression analysis based on the function DPF = exp(-micro(P) * plaque thickness -micro(W) * residual wall thickness). RESULTS: Attenuation in case of (192)Ir was less than the measurement uncertainties. For beta-radiation correlation was found by discrimination between calcified and noncalcified plaque. Classifying noncalcified plaque as normal arterial tissue, the regression coefficient was r = 0.845 at micro(P)= 0.5356 mm(-1) and micro(W) = 0.0663 mm(-1). CONCLUSIONS: Vascular brachytherapy with beta radiation in calcified arteries results in significant dose attenuation within the vessel wall, which can be calculated on knowing the vascular morphometry. Thus, plaque thickness should be taken into account in treatment planning and retrospective analyses.

Arteriosclerosis↗

Data-processing method to reduce error coefficients for membrane-based analytical systems. 1. Amperometric-based sensor evaluated for quantification of oxygen.

This paper describes the use of a predictive, curve-fitting method to reduce the effects of experimental variables on results obtained with membrane-based devices. Multipoint data from the transient regions of responses are used with suitable models and curve-fitting methods to predict the signal that would be measured for the system at equilibrium. The resulting equilibrium response usually is much less dependent on experimental variables than the transient responses used to predict it. The approach is evaluated for the membrane-based amperometric electrode for oxygen. Current vs time data are used to predict the equilibrium current expected when oxygen concentrations are the same on both sides of the membrane. Predicted equilibrium currents vary linearly with oxygen concentration. Relative to the more common steady-state method, the sensitivity of the predictive method is about 5-fold higher, the measurement time is about 17-fold shorter and the dependencies on membrane thickness and stirring rate are 125- and 8-fold lower, respectively. Pooled standard deviations (n = 40) correspond to uncertainties in oxygen concentration of about 0.009 mmol L-1.

Chemistry Techniques, Analytical↗

Quantification of triglyceride transport in blood plasma: a critical analysis.

Reliable and precise quantification of endogenous triglyceride transport in man has not been possible with simple means to date. Direct measurement of net splanchnic secretion of triglyceride fatty acids (TGFA) in very low density lipoproteins (VLDL) provides the must unambiguous information, but precision is low. Coupling infusion of labeled fatty acid with sampling of arterial and hepatic venous blood increases precision; however, the contribution of precursors other than plasma free fatty acids (FFA) must be assessed. Measurement of the rate of hydrolysis of plasma triglycerides after displacing lipases into the blood with heparin holds promise as a simple, nonisotopic method, but it has not been carefully validated and heparin itself alters FFA and triglyceride transport. Multicompartmental analysis following pulse injection of labeled fatty acid offers a practical approach, but uncertainties about the number and location of interacting compartments have made it impossible to determine an absolute value for transport. Reinjection of biologically labeled plasma VLDL is impractical for large scale use, and validity of this approach remains uncertain because of heterogeneity of VLDL-triglycerides and their complex metabolic behavior. Methods to label VLDL-triglycerides in vitro deserve more study as does labeling of other components, such as the B-apoprotein. Such approaches will require rigorous comparison with biologically labeled material as well as careful assessment of alterations in kinetic behavior that may occur when VLDL are separated from blood plasma.

Adult↗