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A simulation study comparing the impact of experimental error on the performance of experimental designs and artificial neural networks used for process screening.

Many variables and their interactions can affect a biotechnological process. Testing a large number of variables and all their possible interactions is a cumbersome task and its cost can be prohibitive. Several screening strategies, with a relatively low number of experiments, can be used to find which variables have the largest impact on the process and estimate the magnitude of their effect. One approach for process screening is the use of experimental designs, among which fractional factorial and Plackett-Burman designs are frequent choices. Other screening strategies involve the use of artificial neural networks (ANNs). The advantage of ANNs is that they have fewer assumptions than experimental designs, but they render black-box models (i.e., little information can be extracted about the process mechanics). In this paper, we simulate a biotechnological process (fed-batch growth of baker's yeast) to analyze and compare the effect of random experimental errors of different magnitudes and statistical distributions on experimental designs and ANNs. Except for the situation in which the error has a normal distribution and the standard deviation is constant, it was not possible to determine a clear-cut rule for favoring one screening strategy over the other. Instead, we found that the data can be better analyzed using both strategies simultaneously.

Culture Media↗

Propagation of population pharmacokinetic information using a Bayesian approach: comparison with meta-analysis.

We investigated the propagation of population pharmacokinetic information across clinical studies by applying Bayesian techniques. The aim was to summarize the population pharmacokinetic estimates of a study in appropriate statistical distributions in order to use them as Bayesian priors in consequent population pharmacokinetic analyses. Various data sets of simulated and real clinical data were fitted with WinBUGS, with and without informative priors. The posterior estimates of fittings with non-informative priors were used to build parametric informative priors and the whole procedure was carried on in a consecutive manner. The posterior distributions of the fittings with informative priors where compared to those of the meta-analysis fittings of the respective combinations of data sets. Good agreement was found, for the simulated and experimental datasets when the populations were exchangeable, with the posterior distribution from the fittings with the prior to be nearly identical to the ones estimated with meta-analysis. However, when populations were not exchangeble an alternative parametric form for the prior, the natural conjugate prior, had to be used in order to have consistent results. In conclusion, the results of a population pharmacokinetic analysis may be summarized in Bayesian prior distributions that can be used consecutively with other analyses. The procedure is an alternative to meta-analysis and gives comparable results. It has the advantage that it is faster than the meta-analysis, due to the large datasets used with the latter and can be performed when the data included in the prior are not actually available.

Administration, Oral↗

Isolation of protein subunits by coupling to an insoluble matrix: analysis of interactions and application to G-actin.

A criterion has been devised for the assessment of intermolecular contacts between protein subunits coupled to a gel matrix. After reversible coupling by way of disulfide groups to Sepharose 4B, the system is exposed to a bifunctional reagent. The protein is then released by reduction, and the proportion of dimers and higher oligomers formed by cross-linking is determined by gel electrophoresis, followed by densitometry of the stained gels. Experiments were performed with G-actin coupled to dithio-2-dipyridyl Sepharose 4B. At low concentration of coupled protein, no species other than the monomer were obtained; under these conditions any intermolecularly cross-linked species would represent pre-existing associated states coupled to the matrix. At higher protein concentrations dimers and higher species progressively appear. Their proportion is much higher than predicted on the basis of a random statistical distribution of coupled molecules throughout the accessible internal volume of the matrix. It follows that protein-protein contacts can occur either because of high flexibility of the polysaccharide chains of the matrix, or because the protein is partly (but not wholly) present as a shell on the surface of the beads. With Affi-gel 10, which has N-hydroxysuccinimide ester coupling groups, similar experiments were performed, taking advantage of the degradation of the matrix under relatively mild acid conditions. In this case the degree of cross-linking was much lower than in the Sepharose 4B system at the same protein concentration. However, this medium proved unsatisfactory for the measurement of interactions of the bound actin with other muscle proteins present in the mobile phase. The results with Sepharose 4B support the validity of previous studies on interaction of monomeric actin with other muscle proteins.

Actins↗

Fluorescence quenching in lecithin and lecithin/cholesterol liposomes by parmagenetic lipid analogues. Introduction of a new probe approach.

1. Perylene, whether incorporated into lecithin or lecithin/cholesterol (1:1) liposomes, exhibits identical fluorescence spectra, but fluorescence in the presence of cholesterol is enhanced by 30-50%. 2. The fluorescence of perylene in pure dipalmitoyllecithin vesicles increases sharply at the transition temperature (Tt equals 41 degrees C). No such fluorescence jump is observed in lecithin/cholesterol (1:1) micelles. 3. In lecithin liposomes maximal quenching of perylene fluorescence at 25 degrees C is effected by cholestane spin label (80%) followed by androstane spin label (70%), 5-nitroxide stearate (60%) and 16-nitroxide stearate (50%). 4. In liposomes containing 5 mol % cholesterol these differences are reduced; however, the sequence of quenching efficiencies is the same except for the nitroxide stearates, which interchange their positions. 5. 5. Paramagnetic quenching of perylene fluorescence is stable below 35 degrees C and above 45 degrees C, but decreases sharply about the phase-transition temperature of dipalmitoyllecithin. 6. In lecithin/cholesterol (1:1, molar ratio) lipsomes fluorescence quenching diminishes linearly, but only slightly, with increasing temperature. 7. Cholestane spin label and androstane spin label at concentrations of greater than 20 mol % themselves suppress the quenching discontinuity at Tt, indicating a cholesterol-like structural effect. 8. The quenching phenomena observed are attributed to a non-random accommodation of fluorophore and quencher molecules (co-clustering) below the phase transition and a statistical distribution of both impurities above Tt. 9. In the presence of cholesterol the clustering tendencies are reduced or even eliminated; this is compatible with the concept that cholesterol fluidizes the phosphatide acyl chains below the transtion temperature.

Androstanes↗

Contribution to the biophysics of the lethal effects of electric field on microorganisms.

The proposed model assumes that the criteria leading to the lethal breakdown of microorganisms suspended in a continuous medium depend on two parameters: (a) the applied electric field must exceed the critical field of membrane to create holes and (b) the Joule energy (deposited in the membrane) must exceed the minimum value beyond which the cell can not recover. The first parameter initiates (reversible) breakdown and the second one, the completion of the (irreversible) electrical breakdown leading to death of the cell. The number of cells surviving the electric field treatment is related to statistical distribution of cell size. Comparison between theory and the experimental results of Kinosita and Tsong (1977); Hülsheger et al. (1980, 1981, 1983); Rosemberg and Korenstein (1990) and others is given.

Bacterial Physiological Phenomena↗

Measurement of signal period on a personal microcomputer and its application to the analysis of cardiac interval and blood pressure.

Personal microcomputers interfaced with analog-digital converters are capable of analyzing almost any biological signal, yet are limited in their ability to directly measure a signal's period/frequency. We describe a short program which measures the interval between periodic events up to 3.5 Hz. In addition, an expanded program which calculates the frequency distribution, distribution statistics, and cross regressions of cardiac interval and blood pressure is described.

Analog-Digital Conversion↗

Scalp recorded somatosensory evoked potentials to posterior tibial nerve stimulation in humans.

The somatosensory evoked potentials (SEPs) produced by stimulation of the right and left posterior tibial nerves individually and also by their simultaneous stimulation were recorded in 84 adult normal subjects up to 150 msec after the stimulus by electrodes placed on the cranial vertex and by rows of electrodes over the sagittal and coronal lines using references on the ear or in the nasopharynx. The statistical distribution of the latencies of their different peaks was established. The effect of simultaneous stimulation of right and left posterior tibial nerves on the early SEP components was described. Some details of the anatomy of the rolandic sulcus were inferred from the amplitude distribution of these potentials.

Adult↗

Preclustered receptor arrangement is a prerequisite for galactose-specific clearance of large particulate ligands in rat liver.

We had hypothesized that preclustered arrangement of galactose-specific receptor activity on rat liver macrophages enables these cells to internalize multivalent, particulate ligands in contrast to the clearance of molecules mediated by statistically distributed receptors on hepatocytes. We now took advantage of the nonclustered receptor distribution in newborn rat liver macrophages to study the in vivo clearance of particulate ligands. Gold particles 5, 17, and 50 nm in diameter (Au5, Au17, Au50), coated with lactosylated bovine serum albumin (LacBSA), were injected into the vena cava and livers were perfusion fixed after allowing for binding and uptake for 3 min. In sinusoidal cells from rats 15 days old LacBSA-Au5 and LacBSA-Au17 were taken up by endothelial cells and all sizes by liver macrophages. In newborn rat liver no LacBSA-Au50 or LacBSA-Au17 was retained in liver macrophages. Uptake of LacBSA-Au5 by sinusoidal cells was significant. LacBSA-Au17 was taken up in significant amounts by endothelial cells of newborn rats which correlates to the findings that galactose-specific binding sites on endothelial cells were found to localize as clusters over coated pits irrespective of age. These results demonstrate the crucial role of clustered receptors in binding and uptake of larger particulate ligands via this lectin-like binding activity.

Animals↗

5S rRNA sugar-phosphate backbone protection in complexes with specific ribosomal proteins.

5S ribosomal RNA forms stable specific complexes with ribosomal proteins L18, L25 and L5. In this work, interaction of phosphate residues of E. coli 5S rRNA within 5S rRNA-protein complexes has been studied. For this purpose 5S rRNA with statistically distributed phosphorothioate residues has been used for complex formation and the accessibility of phosphorothioates to iodine cleavage in the complex and in the free state has been studied. In free 5S rRNA, the phosphate residue at A73 was partially protected, probably due to being involved in the organization of the spatial structure of 5S rRNA. This protection is stronger in the complex with three proteins when the 5S rRNA structure is stabilized. In the 5S rRNA-L18 complex only two phosphate groups, G7 and A34, were protected. L25 in a complex with 5S rRNA protects large numbers of phosphorothioate groups concentrating in two clusters, indicating the possibility of two binding sites for this protein on 5S rRNA. The protection pattern differs from that for individual proteins because of the possible rearrangement of the structure.

Base Sequence↗

One parameter model for error in instantaneous centre of rotation measurements.

Many statistical distributions have a single parameter which describes its spread. In the kinematic study of the instantaneous centre of rotation, error in data cause a non-symmetrical distribution for the centre. As an aid in describing its spread, the analytical form of the probability density function was reduced to a single-parameter model. This results in a convenient way of analysing experimental data through the use of an 'ICR Error Chart'.

Biomechanical Phenomena↗

Joint forces in the human pelvis-leg skeleton during walking.

For the calculation of the forces in the hip, knee and ankle joints during walking the knowledge of the three-dimensional movements of the human body and of the forces between foot and ground is a prerequisite. It is shown how this information may be obtained and what accuracy is obtainable. For the calculation of the statically indeterminate system of the lower limbs, consisting of muscles, bones and joints an optimization method is applied. The optimization criterion is the minimization of the muscle forces. Measurements were taken with seventeen male and five female persons. The maximum joint forces are plotted against gait speed, body weight and body size. In addition some statistical distributions are presented.

Adult↗

Relative friction minimization in fixed orthodontic bracket appliances.

The biomechanical and mathematical analysis of friction on an arch wire/bracket combination and the wire supports has demonstrated that there is an optimal relationship, i.e. minimal friction forces, between the bracket width and the distance between the adjacent brackets on the neighboring teeth. With this optimal situation, less force will develop for a given deflection of the arch wire. This, in turn, will lead to fewer frictional effects when the friction forces are assumed to be proportional to the concentrated perpendicular forces on the arch wire. As long as the contacting surface can be regarded as a point, Coulomb's law of friction can be applied in its simplest form. Relative friction minimization can thereby be qualitatively and quantitatively obtained by linear theory for any specified angulation, wire properties and coefficients of friction. Literature stating that narrow brackets will lead to less friction for specified angulation cannot be confirmed according to the presented analysis. The other extreme statement 'Use the widest bracket possible' also cannot be used in principle. Since the analysis shows a mathematical relationship between most of the relevant design variables of an arch wire/bracket combination and the magnitude of friction forces, empirical results can be sorted and ranked by their physical figures instead of by statistical distribution. In addition, it is possible to make recommendations about the appropriate bracket sizes for various cases. The dependences of the coefficients of friction themselves on material, surface roughness, lubrication, etc. are not investigated here. These factors are also very relevant, especially in reduction of the absolute magnitude of frictional forces, but this problem is left to tribologists.

Biomechanical Phenomena↗

Free thiol groups and labile disulfide bonds in the IgG fraction of human serum.

The IgG fraction was isolated from freshly taken blood serum of health persons (male and female) by column chromatography on QAE-Sephadex. After 30 min incubation with DTNB (5,5'-dithio-(2,2'-dinitro)-benzoate) the average photometrically determined quantity of thio-anions was 0.24 +/- 0.02 SH/mole IgG. Since this result remained unchanged even after 24 h incubation with DTNB, interaction with masked thiol groups cannot be assumed. If, however, the serum was incubated with DTNB for 24 h and the IgG fraction then isolated and treated with thioglycolate, an average of 1.51 +/- 0.39 moles of thio-anions were liberated per mole of IgG. This indicates that on 24 h interaction with IgG, DTNB not only reacts with free SH groups, but also opens S-S bonds by a disulfide exchange reaction. The amount of thio-anions resulting from such opened disulfide bonds was calculated as the difference between 1.51 0.24, i.e., 0.64 S-S/mole IgG. This would be accounted for if approximately 54% of the IgG fraction is composed of a subfraction containing 1 labile disulfide bone per mole. The average of 1.51 thio-anions per mole IgG resulted from 130 single values with an essentially normal statistical distribution and standard deviation well within the usual biological range.

Chemical Fractionation↗

A simple test for the random arrangement of muscle fibres.

The random arrangement of a given muscle fibre class has been assessed by estimating 'mean cluster size' in transverse sections of skeletal muscle. The method was found to be useful when the proportion of the fibre class of interest was low. The statistical distribution of this measure was investigated by computer simulation using a hexagonal lattice model of muscle fibre arrangement. An approximate significance test was developed by considering the extreme points of the distribution. Minor changes to the hexagonal lattice model were incorporated to give a more realistic representation of fibre arrangement and these were found to give very similar results to the simpler model.

Computer Simulation↗

Theory of air pollution damage on human populations.

Existing ideas of dose or exposure are discussed and criticized from the view point of the health damage caused by time dependent air pollutant concentrations. The idea of "dosage" is introduced and some of its mathematical properties are shown and illustrated. The problem of whether or not there exists threshold concentration in air pollution, as well as that of the relation between equi-exposure and equi-damage, is then discussed by making use of the ideas of dosage and healing power of the population. The assumption of this discussion is that the statistical distribution of air pollutant concentrations is log-normal, and that its time series is stationary.

Air Pollution↗

Carbon-13 NMR studies of chloroplast membranes: carbon-13 enrichment without 13C-13C couplings.

A novel approach to carbon-13 (13C) enrichment of chloroplast membranes (and plant materials in general) is presented for 13C-nuclear magnetic resonance (13C-NMR) studies. The method minimizes the occurrence of spectral complications arising from 13C-13C couplings resulting from a statistical distribution of 13C within the molecule with low probability of encountering two 13C atoms adjacent to each other. This is achieved by growing the plants in light surrounded by an atmosphere containing 1/3rd 12CO2 and 2/3rd 13CO2, liberated by weak acid-treatment of a mixture of corresponding barium carbonate salts.

Carbon Isotopes↗

A method for simulating the reflex output of a motoneuron pool.

An analysis of the reflex output of a motoneuron pool in response to a Ia-afferent input is presented. The analysis is based upon a model of the motoneuron pool which includes the subthreshold behavior of motoneurons (integration of synaptic inputs) and the statistical distribution of the motoneurons according to their resting conductance. The latter feature allows for the orderly recruitment of the motoneurons in the order of low resting conductance to high resting conductance. The number of active motoneurons (i.e. the excitation level) is determined by the balance of the excitatory and inhibitory conductances acting on the pool. The reflex output in response to a Ia-EPSP is computed at various excitation levels and with different amounts of presynaptic inhibition. The reflex output is the same for a given excitation level, regardless of the mixture of excitatory and inhibitory postsynaptic conductances used to produce that excitation level. In contrast, presynaptic inhibition markedly affects the relation between reflex output and excitation level.

Action Potentials↗

Rate of aging, rate of dying and the mechanism of mortality.

The history of the search for the law and the mechanism of mortality is reviewed. Recent evidence is summarized showing that the Gompertz law of exponentially increasing force of mortality is only an approximate model of mortality kinetics; various other models also provide a more or less satisfactory fit of mortality kinetics data. In particular, a simple model proposed by the author contains the Gompertz model as a special case and is of general validity: it consists of exponentially increasing cumulative mortality in an initial age range followed by exponentially decreasing survivorship. The various proposed mechanisms underlying mortality kinetics are reviewed, with emphasis on their origin and similarities, and a mechanism is proposed mending two basic classical ideas which are only partially valid: (1) Gompertz's accelerated decline of vitality coupled with identical aging rates of the individuals of a population; and (2) Simms' idea of statistically distributed individual aging rates with a uniform average aging rate (linear decline of physiological vitality). This theory provides a basis for analyzing the relationship between rates of aging and rates of dying.

Aging↗