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[Analysis of the clinical decision in the initial management of BPH].

RATIONALE: Diagnostic management of prostate benign hyperplasia remains a controversial issue subject to variations as made evident in surveys conducted in our country showing that 47.9% urologists regularly perform intravenous urography. The aim of this paper was to determine the preferred strategy from the standpoint of a more accurate diagnosis for evaluation of patients with prostate benign hyperplasia and no absolute indication for surgery. MATERIAL AND METHODS: The methodology used was an analysis of the decision taken by elucidation of the problem using a decision tree with three major choices: (a) to perform IPSS, flowmetry and ultrasound: (b) to perform IPSS and flowmetry, or (c) to perform IPSS alone. Basic analysis by estimation of the expected value and three sensitivity analysis, one-tailed and two-tailed, were used to see whether the dominant choice changed. RESULTS: The choice of performing IPSS alone, resulted in accurate diagnosis adjustment in 80.5% cases. When flowmetry was added from the beginning, this percentage declined to 66.2%; if ultrasound was also done, the decline reached down to 11.2%. The Odds Ratio (OR) for diagnosis imbalance was 15.52 and 33, in choice (a) versus choices (b) and (c), respectively. The OR for the IPSS and flowmetry choice to cause imbalance versus IPSS alone was 2, 12. Also, the choice with greater expected value was IPSS, and this result did not change with the sensitivity analysis. CONCLUSIONS: The choice that considers the possibility of symptom quantification in the IPSS scale and, based on the results, continuation of the diagnostic sequence is the one that should be followed, since it provides higher effectiveness from the standpoint of diagnosis adjustment.

Decision Trees↗

HIV viral load: the myth of the undetectable?

There is a wealth of data supporting the use of viral load measurements to monitor therapy. Indeed, clinical drug trial endpoints routinely include the proportion of patients with a plasma viral load reduction of greater than 0.5 log(10), or greater than 1 log(10). Since a higher viral load reflects increased amounts of virus replication, it would seem desirable to reduce this replication as far as possible, so that the goal of therapy has become one of viral undetectability in plasma. However, virological suppression to undetectable levels is not an absolute determinant of outcome because recent observational cohort data suggest that any significant reduction of viral load is associated with clinical benefit. There are also technical problems when attempting to measure undetectability, with lower limits of detection of 400 or 50 RNA copies/ml of plasma being driven more by the performance of commercial assays than by any inherent cut-off value with proven prognostic significance. Furthermore, the obsession with undetectability has created the concept of the 'viral blip', or 'intermittent viraemia' commonly defined as a single viral load measurement of between 50 and 400 copies/ml, preceded and followed by consistent measurements of less than 50 copies/ml, in a patient receiving therapy. Such blips should be considered in the context of frequent transient changes in viral load which occur below the lower limit of detection by existing laboratory assays. In my view, there remains a misunderstanding about the importance ascribed to these relatively minor changes in lower detection limits, when considered against the background of virus within the body as a whole. I also consider other possible uses of HIV-1 quantification in clinical practice, such as identifying the inherent potency of antiviral regimens.

Anti-HIV Agents↗

Concordance of coronary artery calcium estimates between MDCT and electron beam tomography.

OBJECTIVE: The objective of our study was to compare MDCT with electron beam tomography (EBT) for the quantification of coronary artery calcification (CAC). MATERIALS AND METHODS: Sixty-eight patients underwent both MDCT and EBT within 2 months for the quantification of CAC. The images were scored in a blinded fashion and independently by two observers with a minimum of 7 days between the interpretations of images obtained from one scanner type to the other. RESULTS: Presence versus absence of CAC was discordant by EBT versus MDCT in 6% (n = 4) of the cases by observer 1, with one of these cases also discordant by observer 2. All cases except one (aortic calcium misidentified as CAC) were among those with a mean Agatston score of less than 5 present on EBT but absent on MDCT. EBT and MDCT scores correlated well (r = 0.98-0.99). The relative median variability between EBT and MDCT for the Agatston score was 24% for observer 1 and 27% for observer 2 and was 18% and 14%, respectively, for volume score (average for both observers: 27% for Agatston score and 16% for volume score). Scores were higher for EBT than MDCT in approximately half of the cases, with little systematic difference between the two (median EBT-MDCT difference: Agatston score, -0.55; volume score, 3.4 mm3). The absolute median difference averaged for the two observers was 28.75 for the Agatston score and 15.4 mm3 for the volume score. CONCLUSION: Differences in CAC measurements using EBT and MDCT are similar to interscan differences in CAC measurements previously reported for EBT or for other MDCT scanners individually.

Aged↗

Quantification of CD4, CCR5, and CXCR4 levels on lymphocyte subsets, dendritic cells, and differentially conditioned monocyte-derived macrophages.

CCR5 and CXCR4 are the major HIV-1 coreceptors for R5 and X4 HIV-1 strains, respectively, and a threshold number of CD4 and chemokine receptor molecules is required to support virus infection. Therefore, we used a quantitative fluorescence-activated cell sorting assay to determine the number of CD4, CCR5, and CXCR4 antibody-binding sites (ABS) on various T cell lines, T cell subsets, peripheral blood dendritic cells (PBDC), and monocyte-derived macrophages by using four-color fluorescence-activated cell sorting analysis on fresh whole blood. Receptor levels varied dramatically among the various subsets examined and typically varied from 2- to 5-fold between individuals. CCR5 was expressed at much higher levels in CD4+/CD45RO+/CD62L-true memory cells compared with CD4+/CD45RO+/CD62L+ cells. Fresh PBDC had the highest number of CCR5 ABS among the leukocyte subsets examined but had few CXCR4 ABS, affording a strategy for sort-purifying PBDC. In vitro maturation of PBDC resulted in median 3- and 41-fold increases in CCR5 and CXCR4 ABS, respectively. We found that macrophage colony-stimulating factor caused the greatest up-regulation of both CCR5 and CXCR4 on macrophage maturation (from approximately 5,000 to approximately 50, 000 ABS) whereas granulocyte-macrophage colony-stimulating factor caused a marked decrease of CXCR4 (from approximately 5,000 ABS to <500) while up-regulating CCR5 expression (from approximately 5,000 to approximately 20,000 ABS). Absolute ABS for CD4 and the major HIV-1 coreceptors serve as a more quantitative measure of cell surface expression, and we propose that this be used for future studies looking at the modulation of CD4 or chemokine receptor expression by cytokines, HIV-1 infection, or receptor polymorphisms.

CD4 Antigens↗

Quantification of the wake of rainbow trout (Oncorhynchus mykiss) using three-dimensional stereoscopic digital particle image velocimetry.

Although considerable progress has been made within the last decade in experimental hydrodynamic analyses of aquatic locomotion using two-dimensional digital particle image velocimetry (two-dimensional DPIV), data have been limited to simultaneous calculation of two out of the three flow velocity variables: downstream (U), vertical (V) and lateral (W). Here, we present the first biological application of stereo-DPIV, an engineering technique that allows simultaneous calculation of U, V and W velocity vectors. We quantified the wakes of rainbow trout (Oncorhynchus mykiss, 16.5-21.5 cm total body length, BL), swimming steadily in a recirculating flow tank at a slow cruising speed of 1.2 BL s(-1). These data extend the comparative basis of current hydromechanical data on the wakes of free-swimming fishes to the salmoniforms and are used to test current hypotheses of fin function by calculations of mechanical performance and Froude efficiency. Stereo-DPIV wake images showed three-dimensional views of oscillating jet flows high in velocity relative to free-stream values. These jet flows are consistent with the central momentum jet flows through the cores of shed vortex rings that have been previously viewed for caudal fin swimmers using two-dimensional DPIV. The magnitude and direction of U, V and W flows in these jets were determined over a time series of 6-8 consecutive strokes by each of four fish. Although the fish swam at the same relative speed, the absolute magnitudes of U, V and W were dependent on individual because of body size variation. Vertical flows were small in magnitude (<1 cm s(-1)) and variable in direction, indicating limited and variable vertical force production during slow, steady, forward swimming. Thus, in contrast to previous data from sunfish (Lepomis macrochirus) and mackerel (Scomber japonicus), the trout homocercal caudal fin does not appear to generate consistent vertical forces during steady swimming. U flows were of the order of 3-6 cm s(-1); lateral flows were typically strongest, with W magnitudes of 4-6 cm s(-1). Such strong lateral flows have also been shown for more derived euteleosts with homocercal caudal fins. The ratios of the magnitudes of wake flow, U/(U+V+W), which is a flow equivalent to mechanical performance, were also dependent on individual and ranged from 0.32 to 0.45, a range similar to the range of mechanical performance values previously determined using standard two-dimensional DPIV methods for caudal fin locomotion by more derived euteleosts. Strong lateral jet flow appears to be a general feature of caudal fin locomotion by teleosts and may reflect the nature of undulatory propulsion as a posteriorly propagated wave of bending. Froude efficiency (eta(p)) was independent of individual; mean eta(p) was 0.74, which is similar to previous findings for trout.

Animals↗

Mast cells in pathological and surgical scars.

AIM: To investigate the role of mast cells in surgical and pathological scar reactions by their identification and quantification using immunohistochemistry. METHODS: Surgical scars and pathological scar reactions were stained immunohistochemically for tryptase to identify mast cells. These were quantified in the scar tissue and surrounding dermis. Statistical analyses were performed to test the hypothesis that mast cell numbers were different in the varying types of scar reaction. RESULTS: A significant difference was found between the mean number of mast cells in periocular scars compared with keloids, hypertrophic scars, and surgical scars from other sites (p < 0.05). No significant difference was found in mast cell numbers between the other scar types either within the lesions or surrounding dermis. There were significantly more mast cells in the dermis than in the scar tissue itself, except for the small group of periocular scars. The ratio of mast cells in the lesion compared with the dermis was not significantly different between the scar types, except for the periocular scars. CONCLUSIONS: Mast cell numbers are similar in and around keloid, hypertrophic, and surgical scars. The increased number of mast cells at periocular scar sites was contrary to expectation since keloids are rare at this site. Absolute mast cell numbers may not be an accurate measure of tissue concentrations of active mast cell products. Further comparisons between immunological characteristics of keloid and periocular scars may elucidate specific immunological abnormalities of keloid scars, and this has implications for the development of immunotherapy.

Adult↗

Sexual dimorphism in canine shape among extant great apes.

There have been numerous attempts to sex fossil specimens using the canine dentition. Whether focused on canine size or canine shape, most of these efforts share two deficiencies: lack of quantification of male-female differences in the adopted criteria and a failure to adequately explore among extant species the discriminatory power of these criteria. Here, canine shape indices relating to relative canine height, upper canine root/crown proportionality, and relative length of the lower canine mesial ridge were calculated for males and females of all species and subspecies of extant great apes and two species of gibbons. The accuracy of these indices for identifying the sex of the extant ape specimens was investigated through discriminant analysis and the use of bivariate plots of the two upper and two lower canine indices. The indices were found to be highly accurate in identifying the sex of great ape individuals, not only in single-species and subspecies samples but in mixed-species samples as well; assignment error rates were mostly between 0 and 4%. Accuracy was lowest in Pan (error rates as high as 15%) and highest in Pongo (one error). In most cases, error rates were lower in the upper canines. The effectiveness of these shape indices for sexing might be related to the degree of absolute canine size dimorphism; the indices did not effectively segregate males and females among minimally canine-dimorphic gibbons. The mixed-species results reveal that same-sex index values are remarkably concordant across great ape species, as are the patterns of spatial segregation of males and females in the bivariate plots. Results suggest that, while the indices can be used with some confidence to sex individual fossil specimens, their greatest utility will be for identifying the sex of groups of canines united by size and morphology.

Animals↗

Validation of computerized three-dimensional reconstruction of intravascular ultrasound: measurements of absolute luminal diameter and cross-sectional area in ex vivo human coronary arteries.

UNLABELLED: Computer based 3-dimensional reconstruction transforms 2-dimensional intravascular ultrasound images into a longitudinal format facilitating analysis of luminal narrowing. To validate the accuracy of current software in measuring coronary artery diameter and cross-sectional area, in arteries with atherosclerosis, we performed 3-dimensional reconstruction in 10 human pathologic coronary arterial segments of 10-25mm length. Images were obtained using a 4.8 French catheter with pullback speed of 1mm/sec acquired at 3 frames/sec onto VHS tape. The data were digitized and intraluminal 3-dimensional reconstruction performed using a voxel-based program. Pathologic sections were obtained every 3mm, and dimensions were measured with a resolution of 0.01 mm. Maximum, minimum, and 3 other representative diameters were recorded by an observer blinded to the ultrasound diameters. Average histo-pathologic diameters were reported, and specimen cross-sectional area was then calculated. RESULTS: In 53 sections, pathological diameters ranged from 1.4-4.5mm (mean 2.7 +/- 0.68mm) while 3-dimensional reconstructed diameters were 1.9 to 3.8mm (mean 2.6 +/- 0.54mm). Pathologic and ultrasound derived 3-dimensional reconstruction diameters had an excellent correlation (r=0.86, SEE=+/-0.36). Pathology and 3-dimensional reconstruction cross-sectional area also correlated closely (r=0.88, SEE=+/-1.50). Diameters less than 2.0mm were systematically overestimated and diameters greater than 3.5mm underestimated by 3-dimensional reconstruction. Most 3 dimensional reconstruction values were within +/- 10% of pathology, but diverged at each diameter extreme, approaching +/- 20%. Thus, computerized 3-dimensional reconstruction of ultrasound images shows excellent quantification of luminal size in the 2.0-3.5mm range, suggesting important investigative and clinical applications.

Arteries↗

Influence of different fixation procedures on the quantification of infarction and oedema in a rat model of stroke.

In pharmacodynamic studies using focal ischaemia models, the size of the infarct measured by quantitative histology is the most important outcome measure. Precise, unbiased and reproducible assessment of infarct volume is of foremost importance. A frequent problem in interventional stroke models is the evaluation of infarcts in animals found dead, where instant post-mortem fixation of the brain cannot be performed. The purpose of this study was to investigate possible bias from perfusion, immediate and 3-h post-mortem delayed immersion fixation on the measured volumes of cerebral infarction, oedema and hemispheres in a rat embolic stroke model. Thirty-six male Sprague-Dawley rats were thromboembolized into the internal carotid artery. After survival for 24 h, the animals were divided into three groups: group 1 - immediate perfusion fixation; group 2 - immediate immersion fixation of the brain; and group 3 - animals left dead for 3 h at room temperature before removal of the brain for immersion fixation. Following histological preparation and evaluation, the volumes of the hemispheres and infarction were measured by quantitative histology and planimetry. Brains fixed by immersion were 7% larger than the perfusion-fixed brains. Delaying the immersion fixation for 3 h may increase hemisphere volume by a further 12%. Independent of the fixation procedure, the size of infarction was approximately 40% of the ipsilateral hemisphere, and the oedema was approximately 11% of the size of the infarct. The used planimetric technique was accurate with measured values within +/- 2% of the factual value. In conclusion, sizes of hemispheres, infarction and oedema in absolute volume measures are influenced by the effect of unwanted variation of brain size caused by biological factors and artificial shrinkage caused by fixation, dehydration and heat treatment of the specimens. Infarction and oedema expressed relatively in per cent of hemisphere and infarct, respectively, are robust measures independent of the investigated fixation procedures.

Animals↗

Analysis of encainide and metabolites in plasma and urine by high-performance liquid chromatography.

A high performance liquid chromatography (HPLC) method for the quantification of encainide (4-methoxy-2'-[2-(1-methyl-2-piperidinyl)-ethyl]benzanilide hydrochloride) in plasma and urine has been developed and validated. Encainide and two metabolites, the O-demethyl (ODE) and 3-methoxy-O-demethyl (MODE) congeners of the drug, can be quantified simultaneously in plasma and urine by this procedure. The compounds are extracted from buffered plasma (pH 8.5) using N-butyl chloride containing 5% (vol/vol) isopropyl alcohol. The compounds are then separated on a silica column using ethanol:water:methanesulfonic acid, 500/60/0.2 (vol/vol/vol) as the mobile phase and quantified by measuring their UV absorbance at 254 nm. The lower limit of detection for the analytes was 5 ng/ml in plasma and 25 ng/ml in urine. The method was linear from 5 to 5,000 ng/ml for plasma and 25 to 5,000 ng/ml for urine. The intra-assay precision of the method for encainide, ODE, and MODE in plasma and urine ranged from 2 to 8% RSD depending upon concentration. The inter-assay precision of the method was less than 6% for the three analytes per ml of plasma and urine. Absolute recovery of the analytes from plasma ranged from 82 to 92%, while recoveries from urine ranged from 83 to 99%. The analytes were shown to be stable in frozen plasma and urine for up to 52 weeks.

Anilides↗

Accuracy of deconvolution analysis based on singular value decomposition for quantification of cerebral blood flow using dynamic susceptibility contrast-enhanced magnetic resonance imaging.

Deconvolution analysis (DA) based on singular value decomposition (SVD) has been widely accepted for quantification of cerebral blood flow (CBF) using dynamic susceptibility contrast-enhanced magnetic resonance imaging (DSC-MRI). When using this method, the elements in the diagonal matrix obtained by SVD are set to zero when they are smaller than the threshold value given beforehand. In the present study, we investigated the effect of the threshold value on the accuracy of the CBF values obtained by this method using computer simulations. We also investigated the threshold value giving the CBF closest to the assumed value (optimal threshold value) under various conditions. The CBF values obtained by this method largely depended on the threshold value. Both the mean and the standard deviation of the estimated CBF values decreased with increasing threshold value. The optimal threshold value decreased with increasing signal-to-noise ratio and CBF, and increased with increasing cerebral blood volume. Although delay and dispersion in the arterial input function also affected the relationship between the estimated CBF and threshold values, the optimal threshold value tended to be nearly constant. In conclusion, our results suggest that the threshold value should be carefully considered when quantifying CBF in terms of absolute values using DSC-MRI for DA based on SVD. We believe that this study will be helpful in selecting the threshold value in SVD.

Algorithms↗

Microscopic quantification of hypercin fluorescence in an orthotopic rat bladder tumor model after intravesical instillation.

We have previously investigated the possibility of using hypericin as a diagnostic tool for the fluorescence detection of flat bladder carcinoma. In these clinical studies, it was shown that following intravesical application in humans, hypericin becomes selectively localized in transitional papillary carcinoma and carcinoma in situ (CIS). In the present study, we characterized the biodistribution of hypericin in rat bladder tumor and normal bladder layers after intravesical administration. The biodistribution was evaluated using fluorescence microscopy with computerized image analysis to image and quantify the fluorescence of hypericin across the urothelial tumor and normal bladder wall. The results show that the photosensitizer intravesical administration route provides selective labelling of both the tumor and normal urothelium. A hypericin dose and instillation time-dependent increase in the hypericin fluorescence intensity in both the tumor and urothelium was observed, without significant hypericin fluorescence in the submucosa and muscle layers. The highest fluorescence ratios in hypericin accumulation in the tumor and normal urothelium to the muscle layer were achieved at 4 h with 30 micro M hypericin (20:1 for the urothelium to muscle layer and 30:1 for the tumor to muscle layer). The difference in fluorescence intensity in tumor tissue to the muscle layer following instillation of 8 micro M hypericin was 11:1, 25:1 and 28:1 at 1, 2 and 4 h, respectively. The difference in fluorescence intensity in tumor tissue to the muscle layer using 30 micro M hypericin was 17:1, 27:1 and 31:1 at 1, 2 and 4 h, respectively. The highest absolute fluorescence levels were observed in the tumor at 4 h with 30 micro M hypericin instillation. The results suggest that under these conditions, PDT with hypericin is likely to produce uniform urothelial tumor eradication, without causing damage to the underlying muscle layers.

Animals↗

Modelling HIV-RNA viral load in vertically infected children.

Human immunodeficiency virus (HIV) ribo-nucleic acid (RNA) viral load is a measure of actively replicating virus and is used as a marker of disease progression. For a thorough understanding of the dynamics of the evolution of the virus in the early life of HIV-1 vertically infected children, it is important to elucidate the pattern of HIV-RNA viral load over age. An aspect of assay systems used in the quantification of RNA viral load is that they measure values above particular cut-off values for detection, below which the assays used are not sufficiently sensitive. In this way, measurements are potentially left-censored. Recent adult studies suggest that to adequately model RNA pattern over age, it is necessary to account for within-subject correlation, due to repeated measures, and censoring. The aim of this study, therefore, was to establish whether it is necessary to use complex methods to allow for repeated measures within individuals and censoring of the HIV-RNA viral load in children enrolled in a cohort study. The approach involved the identification of an appropriate model for the basic pattern of RNA viral load by age and subsequent assessment of various estimation procedures accounting for repeated measures and censoring in different ways. Methods developed by Hughes involving the expectation-maximization (EM) algorithm and the Gibbs sampler were taken as the benchmark for comparison of simpler alternatives. Other approaches considered involve linear mixed-effects and ordinary least squares in which censoring is dealt with informally by taking the cut-off value as absolute or taking the mid-point between cut-off and zero. Fractional polynomials provided a substantially superior approach for modelling the dynamics of viral load over age compared to conventional polynomials or change-point models. Allowing for repeated measures was necessary to improve the power of the likelihood ratio tests required to establish the final model, but methods beyond taking the mid-point for censored values did not further improve the fit. Although Hughes' methodology is the best approach, its implementation is not necessary for the identification of the optimal model.

Algorithms↗

Multiphasic cardiac magnetic resonance imaging: normal regional left ventricular wall thickening.

Magnetic resonance imaging (MRI) is a completely noninvasive method for visualizing cardiovascular anatomy but has had limited use for assessment of cardiac function. The authors evaluated the use of gated MRI for the quantification of regional myocardial contraction. Nine normal subjects underwent gated MRI of five transverse sections (7 mm thickness) through the left ventricle at five intervals in the cardiac cycle using a new technique called rotating gated sequence. All five sections were examined, and the section that best demonstrated the midportion of the left ventricle in its maximum dimension was used to obtain measurement. This technique permitted assessment of regional wall thickening of various regions of the left ventricle in different phases of the cardiac cycle. The extent and percentage of wall thickening were calculated from measurements of the septum and anterior and lateral left ventricular wall in end-diastole and end-systole. The calculated mean values for extent and percentage of wall thickening for the septum were 0.40 cm and 40%; for the anterior wall, 0.61 cm and 73%; and for the lateral wall, 0.53 cm and 57%, respectively. A limitation of the current technique in wall thickness measurements is that the transverse MR plane of section is not perpendicular to the long axis of the left ventricle. Consequently, such oblique sections through the left ventricle may give inaccurate absolute wall thickness measurements but can provide reliable estimate of regional wall thickening dynamics. The ability to define left ventricular wall thickness and function without contrast media provides a noninvasive technique for the detection of segmental left ventricular myocardial dysfunction in ischemic heart disease.

Adult↗

[Recovery from anesthesia with enflurane and neuroleptanesthesia. Quantitative assessment with the aid of psychopathometric tests].

Methodological differences between techniques used to examine recovery from anesthesia prevent direct comparison of results. Also psychological tests are not validated for certain anaesthesiological considerations. The two tests presented here combine ease of use for a bedside test with the advantage that the results are expressed as absolute values of cerebral depression. Based on Wieck's concept of symptomatic or functional psychosis they were developed to document the course of neuropsychiatric illness. Their application following anaesthesia seems to be justified as anaesthesia may be regarded as being a deliberately induced functional psychosis characterized by extremely compressed dynamics. The target of a first pilot study were 60 patients following neuroleptanalgesia with and without antagonization with naloxone, or enflurane anaesthesia, respectively. Psychopathometric evaluation yielded an obvious post-operational decline in the SKT-scores which clearly remained below the pre-operative values even after three hours. All the patients revealed a severe transient syndrome after thirty minutes, a moderate syndrome after one and two hours and a mild syndrome after three hours. The only exception were patients antagonized with naloxone who initially showed an improvement but who afterwards did not behave any better. Neither duration of anaesthesia nor total dose of drugs administered showed any significant correlation to the extent of postoperative cerebral depression. Being uninfluenced by moderating variables such as age and IQ, the psychopathometric tests presented here seem to provide a sensitive and reliable method for quantification of recovery from anaesthesia.

Adolescent↗

[Changes in lipid availability in Venezuela, 1970-1992].

The authors carry out a research focused on the quantification and analysis of the main changes in feeding and nutrition in Venezuela between 1970 and 1992. Such research started with the review and adjustment of the Food Balance Sheets (Hojas de Balance de Alimentos) elaborated by the Instituto Nacional de Nutrición (National Nutrition Institute) between 1970 and 1979 in order to homogenize them in a methodological way to make them similar to those elaborated by that institution and the Fundación Polar for the 1980-90 period. Estimates were made about the daily and per person availability of food for human consumption (DCH) for 1991 and 1992. This report, a partial product of that research, characterizes the evolution of the lipidic DCH in Venezuela for the 1970-1992 period. This period has been divided in seven stages of the evolution of the total energetic DCH, as this reflects well the course of the daily and per person Food Purchasing Power (PCA); there is a direct and strong functional relationship between these two variables. Along those stages the behaviour of the lipidic DCH is studied and we try to view possible relationships between the evolution of the Venezeluans economical situation and the absolute and relative variations observed in the level and the structure of the lipidic DCH. This structure is analyzed from several points of view: groups of food sources, origin, "visibility", and place of origin. One purpose is to determine also which food groups are mainly responsible for the venezuelan's external lipidic dependence. A general picture of the evolution of the DCH for saturated fatty acids and cholesterol is made, as well as of the variations experienced by the P/S and M/S relationships. It was found that the most dynamic elements, those that can explain a very high percentage of the variations observed in the level and the structure of the lipidic DCH were: the groups of foods of Visible Fats, Milk and dairy products, and Meats; vegetal lipids; vegetal-visible and animal-invisible lipidic fractions; imported lipids or lipids of food products which raw materials were imported (oily raw materials to make oils and edible solid fats, and raw materials to make food for poultry and hogs). The importance of the food groups Visible Fats, Milk and dairy products, and Meats, as sources or saturated fatty acids in the diet of the Venezuelans was made evident (89-91% of the respective total DCH), as well as the importance of the food groups Eggs, Meats, Fish and Seafood, and Milk and dairy products, as sources of cholesterol (82-89% of the available total). It was found that the lipid-originated calories account for less than 30% of the total energetic DCH; saturated fatty acids account for less than 10% of the available calories/person/day; the DCH for cholesterol did not reach the level of 300 mg/p/d; the P/S and M/S relationships remained close to 1. These last four facts are considered favorable for the health of the human being.

Cholesterol, Dietary↗

Quantification of chick lens alphaA- and delta-crystallins in experimentally induced ametropia.

PURPOSE: The role of the lens in experimentally induced ametropia is not known. A recent study of the chick lens demonstrated optical quality deterioration with the induction of refractive errors, without alteration in lens morphology, size or shape. A change in lens gradient of refractive index (which is dependent on alpha-, beta-, and delta-crystallin concentration and arrangement), could underlie this observation. The purpose of this work was to quantify the concentrations of alphaA- and delta-crystallin in lenses from chick eyes with induced high myopia or hyperopia. METHODS: White Leghorn chicks were unilaterally fitted on the day of hatching either with translucent plastic goggles to induce form-deprivation myopia (n=21) or with +15 D defocus goggles to induce hyperopia (n=14). The ungoggled contralateral eyes were used as controls. The chicks were refracted twice, once on the day of hatching and again seven days later, using streak retinoscopy. On day 7 chicks were sacrificed, lenses decapsulated, and soluble proteins were isolated. Western blot assays were optimized and used to assess crystallin concentration. RESULTS: Analysis revealed no significant difference in alphaA- or delta-crystallin concentration in lenses from eyes induced with form-deprivation myopia and hyperopia as compared to their respective control eyes. Analysis of the difference in medians of delta-crystallin between the control and treated groups of the myopia and hyperopia experiments revealed significance (p=0.030). CONCLUSIONS: This study suggests that with the induction of ametropia, the increased lens spherical aberration previously noted is not due to a change in the absolute concentration of lens alphaA- or delta-crystallin. However, results suggest that the myopic and hyperopic treatments had different effects on lens delta-crystallin concentration. Further investigation is necessary to expand the current knowledge of the role played by the lens in experimental ametropia.

Animals↗

Quantification of tRNA3243(Leu) point mutation of mitochondrial DNA in MELAS patients and its effects on mitochondrial transcription.

The MELAS syndrome is a mitochondrial encephalomyopathy associated with a point mutation at nucleotide 3243 of mitochondrial DNA (mtDNA). The same mutation has also been found in patients with maternally inherited diabetes mellitus. The mutation occurs within a sequence needed for termination of mitochondrial transcription downstream of the ribosomal RNA (rRNA) genes, thus possibly reducing rRNA synthesis in relation to more distal transcripts. This study presents a family in which maternally transmitted diabetes and MELAS syndrome overlap, and a suggestive correlation between the amount of mutant mtDNA and clinical symptoms is observed. Mutant mtDNA was quantified in several tissues of a newborn infant and the highest amount of mutant mtDNA was found in the placenta, which is promising for the development of genetic counselling in MELAS. The consequences of the MELAS mutation were further studied in cultured clonal myoblasts. We found that the myoblasts with 93% of mutant mtDNA terminate the mitochondrial transcription, resulting in a steady-state amount of 16S rRNA 45 times as high as the more distal transcripts. However, myoblasts with a deletion of mtDNA not involving the transcription termination site had 120 times as much 16S rRNA as the distal transcripts. In both the MELAS myoblasts and in those with a deletion of mtDNA the amount of 16S rRNA increased as the mutant mtDNA increased, suggesting that the production of ribosomal RNAs is a response to the translational defect caused by the mutation. We present evidence here that the MELAS mutation causes a defect in transcription termination, thus leading to no absolute deficiency of ribosomal RNAs, but to a reduced capacity to compensate the defective translation.

Adolescent↗