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PubMed1995

Expressing kidney uptake during 99Tcm-MAG3 renography in terms of administered activity.

In a study of gamma-camera renography, a digitized time-activity curve, or renogram, was standardized in terms of absolute renal uptake. The algorithm for standardization of the renogram was based on a gamma-camera uptake method employed by Gates and Schlegel et al. All count rates for the background-corrected renograms were converted to percent renal uptake per injected dose per minute (%RU). The maximum value for the scale on the y-axis was set at 20% RU. The renograms were generated from 66 kidneys in 35 patients (27 males, 8 females) aged 19-80 years. Using the newly developed computer program, the shape of the renograms was considered abnormal between 8/52 (15%) and 10/53 (19%) kidneys, depending on the observer, all of which were classified as normal when displayed conventionally. All the renograms for 4 patients with one poorly functioning kidney, and between 8 (28%) and 10 (32%) renograms for 31 patients with two poorly performing kidneys, exceeded the maximum g-axis value of 20% RU. Expression of the renograms based on absolute uptake is helpful in avoiding the potential problem of two poorly functioning kidneys, which may be considered near-normal in terms of count rate. The maximum value on the y-axis in 99Tcm-MAG3 renography should be fixed between 25 and 30% RU, while the actual amplitude of the standardized renogram may depend on the method used for quantification.

Adult↗

PubMed1995

Quantification of normal cell death in the rat retina: implications for clone composition in cell lineage analysis.

Naturally occurring cell death complicates the analysis of cell lineage studies by making the surviving members of a clone appear more closely related than they actually are. Here we ask how much normal cell death occurs during rat retinal development, and whether that amount of death is sufficient to confuse the analysis of cell lineage relationships. We measure total cell death in the retina by combining relative counts of dead cells with absolute measurements of total cell loss. For most cell types, but not rods, we find that half of the cells generated die during normal retinal development. We use a computer model to quantify the effects of different amounts of cell death in a simulated lineage study. The simulation indicates that 50% cell death means that clonal variability analysed after the cell death period is not necessarily a good indicator of how much variability actually occurs in the underlying lineage.

Animals↗

PubMed1997

Quantification of loss of haemoglobin components from the circulating red blood cell in vivo.

Previous studies have shown that a considerable amount of haemoglobin is lost from the intact red cell during its lifespan. The aim of this study was to determine the relative contribution of all the haemoglobin components to this process. Therefore, the relative amount of haemoglobins A0, A2, F and the glycated haemoglobins were determined in 24 fractions of different cell age. These fractions were obtained by the combination of counterflow and density centrifugation. When the absolute amount of all haemoglobin components were calculated using the MCH-values of each fraction, it appeared that the mean red cell loss of haemoglobins A0, A2, F, an unknown X and "rest" comprised, respectively, 440, 23, 1, 4 and 1 amol per cell, while the mean gain of the glycated haemoglobins was 84 amol per cell. This resulted in a net loss of 385 amol of haemoglobin per cell. One of the glycated haemoglobins (HbA1e2) turned out to be the product of further carbamylation. It was concluded that in the first half of the red cell lifespan HbA0 and HbA2 decreased by glycation and carbamylation and that in the second half some of the HbA0 and HbA2 but also some of the glycated and carbamylated haemoglobin components leave the red cell. The total loss amounted to about 20%.

Cell Separation↗

PubMed1994

Maternal ingestion of tyrosine during rat pregnancy modifies the offspring behavioral lateralization.

It was previously reported that oral administration of tyrosine (500 mg/kg) to pregnant rats increases tyrosine and monoamines level in the fetal brain and modifies locomotion during postnatal life. In the present study, it was found that this treatment alters behavioral lateralization in the offspring. Neonatal rats whose mothers received tyrosine during the second half of gestation showed a low level of absolute and population laterality in both tail and head movements. The alteration of behavioral lateralization was also found during postnatal development and during adulthood. The T-maze behavioral ontogeny was different for tyrosine-mother and sham-treated or untreated mother rats. During adulthood, the T-max lateralization after stress sessions (a procedure that decreases alternation behavior and facilitates the quantification of behavioral lateralization) was also different in control and tyrosine-mother groups. Neonatal and adult rats showed an increase in right-side movements probability. These data provide evidence that maternal ingestion of a catecholamine precursor during gestation may induce a long-lasting modification of the behavioral lateralization of the offspring.

Administration, Oral↗

PubMed2007

Quantification of low-expressed mRNA using 5' LNA-containing real-time PCR primers.

Real-time RT-PCR is the most sensitive and accurate method for mRNA quantification. Using specific recombinant DNA as a template, real-time PCR allows accurate quantification within a 7-log range and increased sensitivity below 10 copies. However, when using RT-PCR to quantify mRNA in biological samples, a stochastic off-targeted amplification can occur. Classical adjustments of assay parameters have minimal effects on such amplification. This undesirable amplification appears mostly to be dependent on specific to non-specific target ratio rather than on the absolute quantity of the specific target. This drawback, which decreases assay reliability, mostly appears when quantifying low-expressed transcript in a whole organ. An original primer design using properties of LNA allows to block off-target amplification. 5'-LNA substitution strengthens 5'-hybridization. Consequently on-target hybridization is stabilized and the probability for the off-target to lead to amplification is decreased.

DNA Primers↗

PubMed2005

Synthesis of an estrogen receptor beta-selective radioligand: 5-[18F]fluoro-(2R,3S)-2,3-bis(4-hydroxyphenyl)pentanenitrile and comparison of in vivo distribution with 16alpha-[18F]fluoro-17beta-estradiol.

Estrogen receptor beta (ERbeta), a less active ER subtype that appears to have a restraining effect on the more active ERalpha, could be a factor that determines the level of estrogen action in certain estrogen target tissues. ERbeta is found in breast cancer, and its levels relative to ERalpha decline with disease progression. Thus, the independent quantification of ERalpha and ERbeta levels in breast cancer by imaging might be predictive of responses to different hormone therapies. To develop an imaging agent for ERbeta, we synthesized a fluoroethyl analogue of DPN (2,3-bis(4-hydroxyphenyl)propanonitrile), a known ERbeta-selective ligand. This analogue, FEDPN (5-fluoro-(2R,3S)-2,3-bis(4-hydroxyphenyl)pentanenitrile), has an 8.3-fold absolute affinity preference for ERbeta. [18F]Fluoride-labeled FEDPN was prepared from a toluenesulfonate precursor, which provided [18F]FEDPN with a specific activity greater than 3100 Ci/mmol after HPLC purification. Biodistribution studies in immature female rats using estradiol as a blocking agent revealed specific uptake of [18F]FEDPN in the uterus and ovaries. Experiments using ERalpha- and ERbeta-knockout mice demonstrated the expected ERalpha-subtype dependence in the tissue uptake of the known 16alpha-[18F]fluoro-17beta-estradiol ([18F]FES), which has a 6.3-fold preference for ERalpha. The tissue uptake of [18F]FEDPN in the ER knockout mice showed some evidence of mediation by ERbeta, but the levels of specific uptake of this agent were relatively modest. Based on our results, imaging of ERalpha can be done effectively with [18F]FES, but imaging of ERbeta will likely require agents with more optimized ERbeta binding affinity and selectivity than [18F]FEDNP.

Animals↗

PubMed2001

Quantification of all fetal nucleated cells in maternal blood between the 18th and 22nd weeks of pregnancy using molecular cytogenetic techniques.

Different types of nucleated fetal cells (trophoblasts, erythroblasts, lymphocytes, and granulocytes) have been recovered in maternal peripheral blood. In spite of many attempts to estimate the number of fetal cells in maternal circulation, there is still much controversy concerning this aspect. The numbers obtained vary widely, ranging from 1 nucleated cell per 104 to 1 per 109 nucleated maternal cells. The purpose of our project was to determine the absolute number of all different types of male fetal nucleated cells per unit volume of peripheral maternal blood. Peripheral blood samples were obtained from 12 normal pregnant women known to carry a male fetus between 18 and 22 weeks of pregnancy. Three milliliters (3 ml) of maternal blood has been processed without any enrichment procedures. Fluorescence in situ hybridization (FISH) and primed in situ labeling (PRINS) were performed, and fetal XY cells were identified (among maternal XX cells) and scored by fluorescent microscopy screening. The total number of male fetal nucleated cells per milliliter of maternal blood was consistent in each woman studied and varied from 2 to 6 cells per milliliter within the group of normal pregnancies. The number of fetal cells in maternal blood, at a given period, is reproducible and can therefore be assessed by cytogenetic methods. This confirms the possibility of developing a non-invasive prenatal diagnosis test for aneuploidies. Furthermore, we demonstrate that it is possible to repeatedly identify an extremely small number of fetal cells among millions of maternal cells.

Adult↗

PubMed1990

Assessment of a contact-plate sampling technique and subsequent quantitative bacterial studies in atopic dermatitis.

The aerobic bacterial flora of 20 patients with atopic dermatitis and 19 control subjects was extensively sampled using the Williamson and Kligman scrub technique and a contact-plate method (cysteine lactose electrolyte deficient media). Comparison of the two quantitative techniques showed that the contact plate is a reliable and convenient alternative to the scrub technique for the quantification of Staphylococcus aureus, micrococci and coagulase negative staphylococci. Quantification of bacterial flora using both techniques showed high rates of colonization by S. aureus on both involved and clinically normal skin of patients with atopic dermatitis. A linear increase in S. aureus counts with increasing severity of dermatitis was found. In contrast, diphtheroids showed a trend of decreasing isolation rates and counts as the severity of the dermatitis increased. Isolation rates and absolute counts for micrococci/coagulase negative cocci were unaffected by the severity of the dermatitis.

Adult↗

PubMed2006

Preoperative grading of gliomas by using metabolite quantification with high-spatial-resolution proton MR spectroscopic imaging.

PURPOSE: To evaluate proton magnetic resonance (MR) spectroscopic imaging with high spatial resolution for preoperative grading of suspected World Health Organization grades II and III gliomas. MATERIALS AND METHODS: Institutional ethics committee approval and informed consent were obtained for control subjects but were not required for the retrospective component involving patients. Twenty-six patients (10 women, 16 men; mean age, 37.5 years) suspected of having gliomas and 26 age- and sex-matched control subjects underwent proton MR spectroscopy. Absolute metabolite concentrations for choline-containing compounds (Cho), creatine (Cr), and N-acetylaspartate (NAA)-N-acetylaspartylglutamate (total NAA [tNAA]) were calculated by using a user-independent spectral fit program. Metabolic maps of Cho/tNAA ratios were calculated, segmented, and used for MR spectroszpcopy-guided stereotactic brain biopsy. Two-sided paired Student t tests were used to test for statistical significance. RESULTS: Significantly lower Cho levels (P = .002) and higher tNAA levels (P = .010) were found in grade II tumors (n = 9) compared with grade III tumors (n = 17). The average Cho/tNAA ratio over the voxels in the tumor center showed a distinct difference (P < .001) between grade II and III gliomas at a threshold of 0.8 (with ratios <0.8 for grade II). The maximum Cr concentration in the tumor showed a clear-cut threshold between grade III oligodendrogliomas and oligoastrocytomas (Cr level, <7 mmol/L) and grade III astrocytomas (Cr level, >7 mmol/L; P = .020). Comparison between the histopathologic findings from the MR spectroscopy-guided biopsy samples (76 biopsies from 26 patients) and molar metabolite values in corresponding voxels located at the biopsy sampling points showed a negative linear correlation for tNAA (r = -0.905) and a positive exponential correlation for Cho (r = 0.769) and Cho/tNAA (r = 0.885). CONCLUSION: Proton MR spectroscopic imaging with high spatial resolution allows preoperative grading of gliomas.

Adolescent↗

PubMed1998

[Quantification of mitral prosthesis failure by different methods of evaluation by transesophageal echocardiography].

PURPOSE: To evaluate the different methods for grading mitral regurgitation (MR) by transesophageal echocardiography (TEE) in patients with clinical suspicion of mitral prosthesis dysfunction. METHODS: Cardiac catheterization (Cath) was performed in 15 patients for grading the severity of prosthetic MR, divided in two groups based on the presence or absence of severe MR. Prosthetic MR was quantified by TEE using methods commonly used for MR of native valves: subjective assessment by color Doppler, objective assessment based on absolute jet area and on its relative area (jet area/left atrial area) and assessment based on the presence of systolic flow reversal in pulmonary vein. RESULTS: Prosthetic MR was mostly transprosthetic (14 patients) and eccentric (11 patients). There was significant correlation (p < 0.05) between Cath and TEE for identification of severe MR based on subjective assessment and on the presence of systolic flow reversal in pulmonary vein. Identification based on absolute (jet area > 7 cm2) and relative (jet area > 35% of left atrial area) jet areas did not reveal significant correlation with the angiographic grade and showed clear underestimation by TEE when the last method was used. However, there was good correlation (p < 0.05) if relative jet areas > 30% were considered as cut point. CONCLUSION: TEE correctly identified angiographic severe mitral prosthesis regurgitation, mainly by the presence of systolic flow reversal in pulmonary vein and subjective assessment. The estimation of severity of the prosthetic MR by absolute or relative jet area seems to be limited and should be used with caution due to eccentricity of the regurgitant jet, frequently seen in mitral prosthesis dysfunction.

Adult↗

PubMed1997

The ureteric jet index: a novel measure of divided renal function.

The aim of this study was to evaluate an index of divided renal function based on the quantification of the ureteric jets seen on colour Doppler ultrasound of the bladder. Thirty-one patients attending for scintigraphic renography underwent colour Doppler ultrasound with video recording for 5 min. Divided renal function was calculated as the proportion of jets from the right-sided orifice ('jet index'). This was compared with the corresponding 'scintigraphic index' found using Patlak-Rutland graphical analysis. Absolute discrepancies were calculated. Twenty-eight of thirty-one (90%) of studies were diagnostic for the calculation of jet indices. The mean jet index was 52% (n=28, SEM=5.8%) compared to a mean scintigraphic index of 54% (n = 28, SEM = 4.0%). The two scores were correlated, with a correlation coefficient of 0.72 and the median absolute difference between the two scores was 7.7%. Forty-three per cent (12/28) of subjects had discrepancies in the two scores of 5% or less. The score differences, however, showed a highly skewed distribution with 32% (9/28) subjects showing discrepancies over 20%. This discordant group (> 20% difference) included three patients with functional pelviureteric obstruction, one with a pelvic mass and one with an underfilled bladder. Two patients with very poor quality jets had impaired renal function. In one case, the index improved after angioplasty for renal artery stenosis. This simple test is a useful adjunct to urinary tract ultrasound but should be interpreted alongside evidence of renal obstruction, and complements rather than replaces existing tests.

Adolescent↗

PubMed2001

Measurement of cortical dopamine d1 receptor binding with 11C[SCH23390]: a test-retest analysis.

[11C]SCH 23390 is a standard ligand for positron emission tomography (PET) studies on striatal dopamine D1 receptors. Its usefulness for cortical D1 receptor quantification in human PET studies has been questioned but has not been addressed previously. The authors tested the reproducibility of [11C]SCH 23390 binding potential (BP) in cortical areas in five healthy volunteers using three-dimensional PET. Measurement of D1 receptor BP was reproducible in basal ganglia, as well as in all cortical areas studied (intraclass correlation coefficients between 0.81 and 0.92). The absolute variability in cortical areas was 9.21% +/- 0.07%. The reproducibility of cortical D1 receptor BP measurement with [11C]SCH 23390 is equal to that observed with a more recent D1-ligand, [11C]NNC 112. [11C]NNC 112 produces slightly higher specific-to-nonspecific binding ratios but has markedly slower kinetics resulting in a need for a longer scan time. These aspects should be considered when designing studies on the cortical D1-like receptors.

Adult↗

PubMed1999

Quantitative pulmonary single photon emission computed tomography for radiotherapy applications.

Pulmonary imaging using single photon emission computed tomography (SPECT) is the focus of current radiotherapy research, including dose-response analysis and three-dimensional (3D) radiation treatment planning. Improvement in the quantitative capability of SPECT may help establish its potential role in this application as well as others requiring accurate knowledge of pulmonary blood flow. The purposes of this study were to quantitatively evaluate SPECT filtered backprojection (FBP) and ordered subset-expectation maximization (OS-EM) reconstruction implementations for measuring absolute activity concentration in lung phantom experiments, and to incorporate quantitative SPECT techniques in 3D-RTP for lung cancer. Quantitative FBP (nonuniform iterative Chang attenuation compensation, scatter correction, and 3D postreconstruction Metz filtering) and OS-EM implementations were compared with a "clinical" implementation of FBP (uniform multiplicative Chang attenuation compensation and post-reconstruction von Hann filtering), for their ability to improve quantification of inactive and active spherical defects in the lungs of an anthropomorphic torso phantom. Activity concentration estimates were found to depend on many factors, such as region of interest size, scatter subtraction constant (k), postreconstruction deconvolution filtering and, in the case of OS-EM, total number of iterations. In general, reconstruction implementations incorporating compensation for nonuniform attenuation and scatter provided reduced bias relative to the clinical implementation. Potential applications to lung radiotherapy, including dose-functional histograms and treatment planning are also discussed. SPECT has the potential to provide accurate estimates of lung activity distributions that, together with improved image quality, may be useful for the study and prediction of therapeutic response.

Algorithms↗

PubMed2004

The specific uptake size index for quantifying radiopharmaceutical uptake.

Quantitative indices of radionuclide uptake in an object of interest provide a useful adjunct to qualitative interpretation in the diagnostic application of radionuclide imaging. This note describes a new measure of total uptake of an organ, the specific uptake size index (SUSI). It can either be related in absolute terms to the total activity injected or to the specific activity in a reference region. As it depends on the total activity in the object, the value obtained will not depend on the resolution of the imaging process, as is the case with some other similar quantitative indices. This has been demonstrated in an experiment using simulated images. The application of the index to quantification of dopamine receptor SPECT imaging and parathyroid-thyroid subtraction planar scintigraphy is described. The index is considered to be of potential value in reducing variation in quantitative assessment of uptake in objects with applications in all areas of radionuclide imaging.

Adenoma↗

PubMed2005

Comparison of commercial real-time PCR assays for quantification of Epstein-Barr virus DNA.

Clinical research suggests a role for viral load measurement in predicting and monitoring Epstein-Barr virus (EBV)-associated diseases. The aim of this study was to assess the performance of the recently commercially available quantitative assays for EBV based on real-time PCR: the RealArt EBV LC PCR kit and the LightCycler EBV quantification kit. A total of 87 samples were analyzed: 67 samples were obtained from transplant recipients and patients with EBV-associated diseases, 8 samples were obtained from the Quality Control for Molecular Diagnostics 2002 EBV Proficiency Program, and 12 negative qualitative nested PCR samples were used as negative controls. Inter- and intra-assay variabilities were determined by running replicates of two samples. All samples were run in a LightCycler instrument. The differences between positive and negative results were not considered statistically significant (P = 0.5355). There were no false-positive results using either method for nested PCR negative-control samples. The difference in viral load values using the two different methods was considered statistically significant (P < 0.01). The logarithmic linear correlation for both assays was low (r = 0.449) but significant (P < 0.01). The LightCycler EBV quantification kit showed a wider dispersal in results but produced substantially more-accurate melting temperature profile curves. The bias towards lower measurements was considerable in comparison with higher viral load. The differences in PCR efficiency and the presence of mutations could explain the disparity between the two methods. It was concluded that confidence intervals would be required to report the results rather than plain absolute values of viral load for patient monitoring.

DNA, Viral↗

PubMed2000

The effects of breed and level of nutrition on whole-body and muscle protein metabolism in pure-bred Aberdeen angus and Charolais beef steers.

Eighteen pure-bred steers (live weight 350 kg) from each of two breeds, Aberdeen Angus (AA) and Charolais (CH), were split into three equal groups (six animals each) and offered three planes of nutrition during a 20-week period. The same ration formulation was offered to all animals with amounts adjusted at 3-week intervals to give predicted average weight gains of either 1.0 kg/d (M/M group) or 1.4 kg/d (H/H group). The remaining group (M/H) were offered the same amount of ration as the M/M group until 10 weeks before slaughter when the ration was increased to H. Data on animal performance, carcass characteristics and fibre-type composition in skeletal muscle are presented elsewhere (Maltin et al. 2000; Sinclair et al. 2000). On three occasions (17, 10 and 2 weeks before slaughter) the animals were transferred to metabolism stalls for 1 week, during which total urine collection for quantification of Ntau-methylhistidine (Ntau-MeH) elimination was performed for 4 d. On the last day, animals were infused for 11 h with [2H5]phenylalanine with frequent blood sampling (to allow determination of whole-body phenylalanine flux) followed by biopsies from m. longissimus lumborum and m. vastus lateralis to determine the fractional synthesis rate of mixed muscle protein. For both breeds, the absolute amount of Ntau-MeH eliminated increased with animal age or weight (P < 0.001) and was significantly greater for CH steers, at all intake comparisons, than for AA (P < 0.001). Estimates of fractional muscle breakdown rate (FBR; calculated from Ntau-MeH elimination and based on skeletal muscle as a fixed fraction of live weight) showed an age (or weight) decline for M/M and H/H groups of both breeds (P < 0.001). FBR was greater for the H/H group (P = 0.044). The M/H group also showed a lower FBR for the first two measurement periods (both at M intake) but increased when intake was raised to H. When allowance was made for differences in lean content (calculated from fat scores and eye muscle area in carcasses at the end of period 3), there were significant differences in muscle FBR with intake (P = 0.012) but not between breed. Whole-body protein flux (WBPF; g/d) based on plasma phenylalanine kinetics increased with age or weight (P < 0.001) and was similar between breeds. The WBPF was lower for M/M compared with H/H (P < 0.001) based on either total or per kg live weight0.75. Muscle protein fractional synthesis rate (FSR) declined with age for both breeds and tended to be higher at H/H compared with M intakes (intake x period effects, P < 0.05). Changing intake from M to H caused a significant increase (P < 0.001) in FSR. The FSR values for AA were significantly greater than for CH at comparable ages (P = 0. 044). Although FSR and FBR responded to nutrition, these changes in protein metabolism were not reflected in differences in meat eating quality (Sinclair et al. 2000).

Age Factors↗

PubMed1990

"CORT-EX:" a program for quantitative analysis of brain SPECT data.

A program was developed to extract from brain SPECT data global as well as regional concentrations of a radiopharmaceutical while allowing for improved subjective evaluation of its distribution. This program was used to process the data obtained from 17 normal subjects, 20 min, 2 hr, and 4 hr after the injection of iodine-labeled iodoamphetamines. The mean absolute cortical uptake at these three time periods was 0.921 (+/- 0.185), 0.803 (+/- 0.107), and 0.748 (+/- 0.103) in arbitrary units (+/- s.d.), respectively. The regional distribution of the tracer became more uniform with time due to an uneven washout rate. The cerebellum was noted to have a very high variability in its uptake and a high washout rate, making it unsuitable as an internal standard for relative quantification. Finally, a repeat study was performed on 10 subjects. No significant difference could be demonstrated in the mean uptake of the group at 2 and 4 hr, however the difference observed in the 20 min uptake values was significant at the p = 0.05 level.

Adult↗

PubMed1993

Quantification of proteinuria: a re-evaluation of the protein/creatinine ratio for elderly subjects.

The difficulties of 24-hour urine collection are well recognized, especially for elderly people for whom a means of estimating protein excretion from a single casual unit sample would be preferable. We compared measurement of total urinary protein estimated by the protein/creatinine ratio with 24-h urine collection and examined the role of the ratio in discriminating between clinically important levels of proteinuria. Although the protein/creatinine ratio appeared to have an excellent correlation with the 24-h collection, the absolute agreement between the two methods was poor. However, the clinical role of the protein/creatinine ratio as a semi-quantitative test of proteinuria remained, provided adjustment for expected creatinine excretion was made.

Aged↗