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PubMed1986

Alterations of energy metabolism in the spontaneously hypertensive rat: a 31P nuclear magnetic resonance study.

We quantified high-energy phosphate metabolites in hypertensive hypertrophied and normal myocardium and monitored temporal changes using the non-invasive 31P nuclear magnetic resonance (NMR) spectroscopy. Hearts from 18 month spontaneously hypertensive rats (SHR) and age-matched Wistar-Kyoto rats (WKY) were perfused with a phosphate-free buffer at 10 cc/min per g and paced at 240 beats/min on a modified Langendorff apparatus. Perfusion pressure, left ventricular pressure (LVP) and dP/dt were recorded and successive 31P NMR spectra were collected during a 24-min baseline period (oxygenated buffer), anoxia (N2-bubbled and glucose-free buffer) until a 70% fall in LVP occurred, and recovery. An aminomethylphosphonate standard, located within the LVP balloon, permitted absolute quantification of myocardial phosphate moieties (including inorganic phosphate (Pi), creatine phosphate (CP) and ATP). During perfusion, SHR hearts demonstrated higher coronary resistance but no significant differences in LVP or dP/dt. Spontaneously hypertensive rat hearts had lower CP, ATP and CP/Pi ratio and showed a faster fall in cardiac function during anoxia, associated with parallel rates of changes in the phosphate moieties.

Animals↗

PubMed1975

Measurement of absolute amounts of antigen-specific human IgE by a radioallergosorbent test (RAST) elution technique.

A technique for the absolute quantification of antigen-specific human IgE is described. It employs elution of a calculable amount of antigen-specific IgE from an allergosorbent-antibody complex by means of alkaline pH treatment, followed by measurement of the IgE content of the eluate with a modified radioimmunosorbent test (RIST). With this method IgE antibody directed against the benzylpenicilloyl determinant of penicillin (BPO) was measured quantitatively in sera from seven penicillin allergic patients. IgE specific for ragweed antigen E was measured in sera from 33 ragweed allergic patients. Values obtained for IgE anti-BPO ranged from 19 to 1806 ng/ml and comprised from 1.3 to 27.5% of total serum IgE. Values of IgE anti-antigen E ranged from 9 to 1807 ng/ml, comprising from 3 to 84% of total serum IgE. Excellent correlation (r = 0.99; p less than 0.001) was obtained for both antigen systems between values determined by the RAST elution technique and by simple RAST assay with interpolation from a reference serum of known specific IgE content as determined by the elution technique may be needed only for primary standardization of reference sera.

Allergens↗

PubMed1989

In vivo assessment of a digital angiographic method to measure absolute coronary artery diameters.

Several techniques exist for the quantification of absolute coronary artery diameters using radiologic methods. An in vivo assessment of a quantitative technique based on direct digitally acquired images was performed by imaging inflated angioplasty balloons (n = 25), balloon catheter shafts (n = 16) and coronary guidewires (n = 20) at the time of coronary angioplasty. After this, the actual size of the objects was determined with a micrometer. Diameters measured by the quantitative digital method had an excellent correlation with the actual diameters (digital diameter = 0.80 [actual diameter] + 0.32; n = 61; r = 0.97; standard error of the estimate = 0.26 mm; p less than 0.001). Moreover, the correlation between interobserver and intraobserver measurements was excellent (r = 0.99 for both, standard error of the estimate = 0.16 mm and 0.09 mm, respectively). However, there was a consistent error present that was related to the size of the object measured. Objects less than 0.5 mm were consistently overestimated and objects greater than 1 mm were usually underestimated by the digital technique, although the actual magnitude of the error was small. Objects less than 0.5 mm in diameter were overestimated by 0.41 +/- 0.11 mm and objects greater than 1 mm were underestimated by 0.23 +/- 0.19 mm. Based on an analysis of the error present, correction algorithms were formulated and tested prospectively using an additional 29 object measurements. This resulted in an improvement in the quantification of the diameters with a smaller magnitude of error. This in vivo assessment suggests that the rapid online assessment of absolute coronary artery diameters is possible, but also demonstrates important errors inherent in this method.

Algorithms↗

PubMed2026

Estimation of chloroplast macromolecular complex copy numbers and subunit stoichiometries during the Chlamydomonas reinhardtii cell cycle.

An unbiased, quantitative view of biomolecules in a living cell is a prerequisite for accurate modeling approaches and informs our understanding of cellular metabolism at scale. In this work, we used the total protein approach (TPA), in which the total protein mass of a given proteomics sample is used as a calibrator for absolute protein quantification, to determine protein abundances during the Chlamydomonas reinhardtii diurnal cycle. We use external, independently measured quantitative markers (metals, pigments) to assess the absolute protein abundances in unlabeled whole cell extracts. We calculate protein abundances in fg cell-1 of 7322 Chlamydomonas proteins, 2266 of which were captured in every time point, including the major proteins involved in the light reactions, photoprotection, proteostasis, and fatty acid metabolism during a cell cycle. As expected, Rubisco large and small subunits are present in a 1:1 stoichiometry, with the large subunit being the most abundant protein in our data set, averaging 5.05 × 106 molecules per cell, reflecting 2.7% of the total protein mass. We noticed that PSII is the most abundant complex involved in the light reactions with 2.08 × 106 complexes per cell. PSI averages 1.75 × 106 complexes per cell and cytochrome b6f averages 0.77 × 106 complexes per cell. The TPA is a robust tool to study proteome dynamics quantitatively, while avoiding artifacts due to biochemical fractionation. Our proteome data set with an unprecedented temporal resolution is a valuable resource to assess protein abundances during the cell cycle in the reference alga Chlamydomonas.

Chlamydomonas reinhardtii↗

PubMed1992

Determination of absolute phosphate metabolite concentrations in RIF-1 tumors in vivo by 31P-1H-2H NMR spectroscopy using water as an internal intensity reference.

The absolute metabolite quantification method of Thulborn and Ackerman [J. Magn. Reson. 55, 357 (1983)] in which the tissue water proton signal is used as an internal intensity standard and its more recent variation in which NMR peak intensities are referenced to that of the natural abundance deuterium signal of water [Li et al., SMRM Abstr. 2, 825 (1988); Song et al., Magn. Reson. Med. 25, 45 (1992) have been implemented to obtain absolute phosphate metabolite concentrations in subcutaneous RIF-1 tumors during untreated growth and following treatment with 5-fluorouracil. The equivalence of these two hydrogen isotopes as intensity standards and the validity of their use in the determination of absolute metabolite concentrations in vivo by NMR has been demonstrated. On matched in vivo and extract tumor samples (n = 5), excellent agreement has been obtained between nucleoside triphosphate concentrations determined by NMR and those derived by HPLC analysis for the control tumors. Following 3 days of untreated growth, absolute concentrations of phosphate metabolites in RIF-1 tumors (n = 10) decreased significantly, except for the Pi concentration which did not vary. For the treated tumors (n = 10) there were no changes in metabolite concentrations except for a decrease in the PCr and, possibly, Pi concentrations. The PCr/Pi ratio in the latter tumors did not change. These observations suggest that changes in absolute metabolite concentrations may be more sensitive indices of response to therapy than changes in metabolite peak amplitude ratios, a parameter commonly used to express in vivo NMR data.

Animals↗

PubMed1991

Absolute cross-sectional area measurements in quantitative coronary arteriography by dual-energy DSA.

Recent studies have emphasized the limitations of conventional coronary angiography. These limitations include the lack of correlation between the severity of coronary stenosis as estimated from coronary angiograms and the actual severity of stenotic lesions measured in postmortem hearts. As a result, attempts have been made to quantitate luminal dimension more precisely. The application of quantitative digital subtraction angiography (DSA) in the assessment of coronary artery lesion dimension has been limited by cardiac and respiratory motion artifacts. We have reported previously on a motion-immune dual-energy (DE) cardiac mode in which kVp and filtration are switched at 30 Hz. To assess the potential advantages of a videodensitometric technique for quantification of absolute vessel cross-sectional area (CSA), three different quantitative coronary arteriography (QCA) algorithms were compared. The three algorithms under comparison were a videodensitometric (V) algorithm, which does not require any geometric assumption for absolute vessel CSA measurement, and videodensitometric (VC) and edge detection (ED) algorithms, which do require the assumption of circular cross-section for CSA measurements. A cylindrical vessel phantom (0.5-4.75 mm in diameter) and a crescentic vessel phantom, producing 25% to 90% area stenosis, were imaged over the chest of a humanoid phantom. The low- and high-energy images were corrected for scatter and veiling glare before energy subtraction. For CSA measurements in crescentic vessel phantoms, the V algorithm produced significantly improved results (slope = 0.87, intercept = 0.51 mm2, r = .95) when compared to the VC (slope = 1.05, intercept = 4.19 mm2, r = .75) and the ED (slope = 1.57, intercept = 5.21 mm2, r = .60) algorithms.

Algorithms↗

PubMed1991

New grading system of cerebral gliomas using positron emission tomography with F-18 fluorodeoxyglucose.

Positron emission tomography (PET) using fluorine-18 (18F)-fluorodeoxyglucose (FDG) has been reported to be a powerful diagnostic and prognostic tool in patients with primary brain tumors. This study was undertaken to compare the prognostic value of: (1) visual grading of [18]FDG uptake in the tumor, (2) the absolute glucose metabolic rate of the tumor (TMRglc), (3) the ratio of glucose metabolism between the tumor and whole brain (T/WB) and (4) between the tumor and contralateral cerebellum (T/CBL). Each of these four parameters was correlated with the survival time in 20 patients with malignant cerebral gliomas. Excellent correlation was obtained with visual grading and reasonably good correlation was obtained with T/WB or T/CBL, but TMRglc alone was only a fair prognostic indicator. Thus, visual grading provides a qualitative analysis and T/WB provides a semi-quantitative analysis neither of which requires arterial blood sampling for quantification of absolute metabolic rates for glucose.

Brain Neoplasms↗

PubMed1990

Quantitative dual-energy coronary arteriography.

Subtraction techniques for digital cardiac imaging have been hampered by misregistration artifacts. The use of dual-energy imaging is being evaluated as a means for reducing these artifacts. Results reported previously indicate that the dual-energy technique may be useful for applications such as exercise ventriculography and general quantification tasks. The purpose of the current study is to investigate the use of dual-energy subtraction imaging for quantitative coronary arteriography. In vivo coronary vessel phantoms (0.2 to 7 mm2 in cross-sectional area) were used to study the potential advantages of tissue suppressed energy subtracted images over unsubtracted images for quantification of absolute vessel cross-sectional area when cardiac motion is present. Estimates of lumen cross-sectional area (N = 20) were determined using videodensitometric analysis of selected energy subtracted and unsubtracted images. Linear regression analysis of measured and actual cross-sectional area showed energy subtracted image data (slope = 1.06, intercept = 0.48 mm2, r = 0.99) to have improved accuracy (P less than .05) and precision (P less than .05) over unsubtracted image data (slope = 1.24, intercept = 1.07 mm2, r = 0.95).

Angiography, Digital Subtraction↗

PubMed1989

Comparison of two interpolative background subtraction methods using phantom and clinical data.

Two interpolative background subtraction methods used in scintigraphy are tested using both phantom and clinical data. Cauchy integral subtraction was found to be relatively free of artefacts but required more computing time than bilinear interpolation. Both methods may be used with reasonable confidence for the quantification of relative measurements such as left ventricular ejection fraction and myocardial perfusion index but should be avoided if at all possible in the quantification of absolute measurements such as glomerular filtration rate.

Glomerular Filtration Rate↗

PubMed1991

Evaluation of two extraction methods for the determination of egg yolk cholesterol.

Controversy concerning egg cholesterol values exists in recent literature due to varying procedures used for cholesterol determination. The purpose of the present study was to investigate the efficacy of direct sample saponification (Method A) versus saponification of a lipid extract (Method B) for analysis of yolk cholesterol. Method A resulted in a value of 19.1 +/- .4 (SE) mg cholesterol/g of yolk for the National Institute of Standards and Technology (NIST) reference (cholesterol in whole egg powder) as compared with the NIST certified value of 19.0 +/- .2 mg/g. Method B resulted in a significantly lower value of 14.6 +/- .5 mg/g. Egg yolk cholesterol values were determined to be 196 +/- 4.2 mg per egg by Method A and 132 +/- 11 mg per egg by Method B. Various amounts (1, .5, .25 g) of yolk cholesterol assayed by either method proportionately decreased cholesterol values as yolk amount decreased; however, Method B consistently resulted in lower yolk cholesterol. These data suggest that both Methods A and B are valid for determining relative differences between treatments; however, the NIST standard data indicate that for quantification of absolute cholesterol values, direct saponification is more accurate. The NIST standard of cholesterol in whole egg powder should be used as a control for comparing cholesterol data regardless of extraction method used.

Animals↗

PubMed1988

Direct immunomagnetic quantification of lymphocyte subsets in blood.

A method is described where superparamagnetic polymer microspheres coated with monoclonal antibodies (MoAb) are used for the direct and fast quantification of the absolute number of cells of various lymphocyte subsets in blood. Blood samples were incubated with microspheres coated with a subset specific MoAb. Using a magnet the microsphere-rosetted cells were isolated and washed. Following lysis of the cell walls to detach the microspheres, the cell nuclei were stained with acridine orange and counted in a haemocytometer using an immunofluorescence microscope. With MoAb specific for CD2, CD4, CD8 and CD19, reproducible absolute counts of the corresponding lymphocyte subsets were obtained which correlated closely with those obtained by an indirect quantification method.

Cell Separation↗

PubMedDate not supplied

Quantification of coronary arterial calcium by dual energy digital subtraction fluoroscopy.

Clinical studies of the heart with fluoroscopy have shown that fluoroscopic visualization of calcium in the coronary arteries is strongly associated with coronary artery disease. However, fluoroscopic detection is limited by its low sensitivity, which is partly due to the interfering background tissue structures and image quantum noise. Moreover, quantification of the absolute amount of calcium in an arterial segment has not been possible. A real-time dual-energy subtraction technique has been investigated as a possible solution to the above problem. In this energy subtraction technique, the kVp and filtration are switched at 30 Hz. In order to assess the potential utility of this videodensitometric technique to quantitate coronary artery calcium, arterial phantoms and excised segments of diseased human arteries were imaged. The low- and high-energy images were corrected for scatter and veiling glare before subtraction. Calcium measurements were made using the tissue-suppressed energy-subtracted images. The estimated calcium phosphate and ashed weights of the calcified arterial segments (N = 20) were highly correlated (slope = 1.04, Intercept = -0.33 mg, r = 0.92).

Calcium↗

PubMed1988

Measurement of absolute myocardial blood flow with H215O and dynamic positron-emission tomography. Strategy for quantification in relation to the partial-volume effect.

An in vivo technique was developed for measuring the absolute myocardial blood flow with H215O and dynamic positron-emission tomography. This technique was based on a new model involving the concept of the tissue fraction, which was defined as the fraction of the tissue mass in the volume of the region of interest. The myocardium was imaged dynamically by positron-emission tomography, starting at the time of intravenous bolus injection of H215O. The arterial input function was measured continuously with a beta-ray detector. A separate image after C15O inhalation was also obtained for correction of the H215O radioactivity in the blood. The absolute myocardial blood flow and the tissue fraction were calculated for 15 subjects with a kinetic technique under region-of-interest analysis. These results seem consistent with their coronary angiographic findings. The mean value of the measured absolute myocardial blood flows in normal subjects was 0.95 +/- 0.09 ml/min/g. This technique detected a diffuse decrease of myocardial blood flow in patients with triple-vessel disease.

Angina Pectoris↗

PubMed2026

Telomere length in patients with non-functional adrenal incidentalomas.

Telomeres maintain genomic integrity during cell replication by preventing chromosomal fusions. Beside genetic influences, telomere length is affected by environmental factors such as oxidative stress and inflammation. These mechanisms also contribute to metabolic syndrome components linked to cellular aging. We aim to evaluate whether telomere length is shortened in patients with non-functional adrenal incidentaloma (NFAI) compared to the control group. This study was designed as a prospective, single-center study. The total of 88 participants included were 44 patients aged between 40 and 60 years with NFAI in our endocrinology clinic and 44 control subjects. An Absolute Human Telomere Lengths Quantification qPCR Assay kit (Nucleotestbio, Budapest, Hungary) was used for analyses. There was no significant difference between the NFAI and control groups regarding age and sex distribution. Telomere length was significantly shorter in the NFAI group (NFAI group: 3.680 ± 1.970 kb; control group: 4.469 ± 1.672 kb; p = 0.046). While no significant difference was found in telomere lengths in subgroup analyses, patients with basal adrenocorticotropic hormone (ACTH) levels <15 pg/mL had significantly shorter telomeres than those with basal ACTH levels ≥15 pg/mL (p = 0.034). A strong positive correlation was observed only between telomere length and ACTH level (p = 0.001). This study demonstrated that telomere length is significantly shortened in NFAI patients. Here, we propose that the underlying cause of telomere length shortening in the NFAI group may be related to increased cardiovascular risk and an elevated inflammatory state, even in the presence of cortisol levels within the normal range.

Humans↗

PubMedDate not supplied

Absolute lung density in experimental canine pulmonary edema.

The noninvasive quantification of pulmonary edema could be of importance in patient management. We have developed a portable Compton-scatter densitometer capable of measuring density in the range of 0.1-1 g/cm3 independently of the chest wall. Change in lung density was investigated in 19 dogs with pulmonary edema induced by a combination of raised left atrial pressure and hemodilution. The increase in lung density correlated with the postmortem assessment of lung water (r = 0.862, p less than 0.01) and with hemodynamic indicators of pulmonary edema (r = 0.749, p less than 0.01). Subtracting the intravascular component of lung density did not improve the correlation with postmortem lung water (r = 0.850, p less than 0.01). The measurement of absolute lung density by a Compton-scatter technique may be a potentially useful method of quantifying pulmonary edema.

Absorptiometry, Photon↗

PubMed2026

Accurate quantification of canine mitochondrial DNA copy number from canine blood and brain samples.

Acute brain injury is difficult to evaluate in veterinary medicine and tools to investigate the potential involvement of mitochondrial involvement are limited. The brain is highly enriched in mitochondria and contains thousands of copies of mitochondrial DNA (mtDNA) per cell, but robust methods for quantifying mitochondrial DNA copy number (mtDNA-CN) in canine tissues are lacking. We describe the development of a quantitative real-time PCR assay for absolute measurement of mtDNA-CN which was validated in canine blood and brain tissue. To minimize amplification of nuclear mitochondrial insertion sequences (NumtS) and repetitive regions, species-specific oligonucleotide primers were designed following in silico genomic filtering. The assay was applied to a small pilot cohort comprising blood samples from dogs with and without acute brain injury (n = 4-6 per group) and cerebral cortex samples (n = 1 per group) to assess feasibility and biological plausibility. In non-brain injury dogs, blood mtDNA-CN ranged from 98 to 288 copies per nuclear genome (mean 193 ± 72), while values in brain-injured cases ranged from 163 to 228 copies per genome (mean 200 ± 33). Cerebral cortex samples exhibited higher mtDNA-CN than blood, consistent with known tissue-specific mitochondrial enrichment. In a single brain-injured case with serial sampling, mtDNA-CN increased over five days. This study presents a validated assay and pilot data for mtDNA-CN quantification in canine samples. While not powered for biomarker evaluation, this method may enable future studies of mitochondrial dynamics in canine brain injury and metabolic disease.

Animals↗

PubMed1975

Liquid chromatography in pharmaceutical analysis IV: determination of antispasmodic mixtures.

Parameters associated with the separation of antianxiety-antispasmodic agents were investigated using high-pressure liquid chromatography. Eight widely prescribed drugs were studied. The compounds were chromatographed on reversed-phase octadecyltrichlorosilane (C18) or diphenyldichlorosilane (phenyl) columns, using mixtures of absolute methanol and distilled water buffered with ammonium dihydrogen phosphate, ammonium acid phosphate, or ammonium carbonate. A mixture of phenobarbital-propantheline bromide was selected to demonstrate the utility of the separation and quantification method. The mixture was chromatographed on a phenyl column, using absolute methanol-aqueous 1 percent ammonium dihydrogen phosphate (60:40) (pH 5.85) at a flow rate of 1.4 ml/min. Each determination can be achieved in approximately 15 min with an accuracy of 1-2 percent.

Atropine↗

PubMed1991

[Quantification of T-lymphocyte subpopulations. Immunological marker in human immunodeficiency virus infections].

The absolute number of CD4+ and CD8+ T cells was counted directly in the blood of 75 healthy controls and 223 individuals infected with human immunodeficiency virus (HIV). The HIV-seropositive individuals were also classified clinically according to the system recommended by the Centers for Disease Control. We observed a pattern of changes in the T cell subset counts of the patient group. This pattern could be defined by five T cell stages ranging from normal T cell subset values to values representing severe T cell immunodeficiency. A close correlation was observed between the immunological and clinical classification of the patients. Quantification of T cell subsets may provide useful information in the follow-up of individuals infected with HIV, particularly in assessing the indications for antiviral therapy, and its effects.

HIV Infections↗