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Kinetic mechanism of chicken liver xanthine dehydrogenase.

The kinetic behaviour of chicken-liver xanthine dehydrogenase (xanthine/NAD+ oxidoreductase; EC 1.2.1.37) has been studied. Steady-state results, obtained from a wide range of concentrations of substrates and products, were fitted by rational functions of degree 1:1, 1:2, 2:2 and 3:3 with respect to substrates, and 0:1, 1:1, 0:2 and 1:2 with regard to products, using a non-linear regression program which guarantees the fit. The goodness of fit was improved using a computer program that combines model discrimination, parameter refinement and sequential experimental design. The AIC and F tests were also used for model discrimination. For comparative purposes, the xanthine/oxygen oxidoreductase reaction was also studied. From the functions which give the maximum improvement, the complete rate equation was deduced. The significance of the terms was stated by the above methods. It was concluded that xanthine dehydrogenase requires a minimum mechanism of degree 1:1 for xanthine, 2:2 for NAD+, 1:1 for uric acid and 1:2 for NADH in the xanthine/NAD+ oxidoreductase reaction. These are the minimum degrees required but a rate equation of higher degree is not excluded.

Animals

Kinetic studies of chicken and turkey liver mitochondrial aspartate aminotransferase.

The kinetic behaviour of chicken liver and turkey liver aspartate aminotransferases (L-aspartate:2-oxoglutarate aminotransferase, EC 2.6.1.1) was studied. Steady-state data were obtained from a wide range of concentrations of substrates and product L-glutamate. The data were fitted by rational functions of degree 1:1, 1:2 and 2:2 with respect to substrates and 0:1, 1:1, 0:2 and 1:2 with regard to product (L-glutamate), by using a non-linear regression program that guarantees the fit. The goodness of fit was improved by the use of a computer program that combines model discrimination parameter refinement and sequential experimental design. It was concluded that aspartate aminotransferase requires a minimum velocity equation of degree 2:2 for L-aspartate, 2:2 for 2-oxoglutarate and 1:2 for L-glutamate. Finally, a plausible kinetic mechanism that justifies these experimental results is proposed.

Animals

[Dynamics of egg maturation in the parasitoid Lixophaga diatraeae (Diptera, Tachinidae) raised in a substitute host, Galleria mellonella (Lepidoptera, Pyralidae)].

The content of the uterus and ovaries of L. diatraeae (an ovolarviparous tachinid fly) was studied by dissecting 121 females 0 to 25 days old. Several size classes of flies (p: small; m: medium; g: big) were used, weighing 10 to 12.5 mg, 12.5 to 13.5 mg, and 13.5 to 16 mg, respectively. The mean number of ovarioles per ovary, correlated to female size, was 12.1 for p as against 12.9 for m + g females. The total mean number of eggs and oocytes changed with female age and weight. For 0, 5, 10 and 15-day old females, this number was 56, 149, 158 and 202, respectively in g flies and 51, 128, 141 and 139, respectively, in p ones. To study egg maturation dynamics, we counted the total number of eggs + oocytes per p female (Y1), the eggs in the uterus (Y2), partially embryonated eggs (Y3), and entirely embryonated eggs (Y4). The experimental results were fitted to various age functions using a non-linear adjustment program. The models fitting each datum best were asymptotic functions of the form, Y = A [1-e-K(x-D)], where A is the asymptotic value, K a speed constant of the studied phenomenon and D age at the beginning of the phenomenon. At emergence time, each female contained a mean of 50 oocytes which passed into the uterus immediately after mating. Egg production continued for at least 10 days after emergence. The rate of embryogenesis seemed to be influenced by the number of eggs in the uterus and/or the age of the fly; it was high for the first eggs then decreased progressively. In our conditions, the first eggs with a planidium ready to hatch appeared at 10 days post-emergence. To obtain a maximum number of larvae in good health, it is better to dissect 12 to 18-day old females. The egg volume increases slightly during embryogenesis.

Animals

Disposition of propofol in patients undergoing surgery. A comparison in men and women.

The disposition kinetics of propofol have been determined in 12 patients (six female) receiving propofol 2.5 mg kg-1 for induction of anaesthesia, which was maintained with 67% nitrous oxide in oxygen and 1-1.5% halothane. Peripheral blood samples were collected at selected times up to 8 h after the injection of the drug, and whole blood propofol concentrations determined by HPLC with fluorescence detection. Drug concentration-time data were analysed by the non-linear regression program ELSFIT. This showed the data to be describable by a tri-exponential equation, corresponding to a three-compartment model. There were no differences in the derived kinetic indices for the male and female patients, with the exception of a greater Vdss:V gamma ratio in the males. The terminal half-life in the male patients was 262 min (SEM 44), and in the female patients 309 min (60). Vdss was 329 litre (67) and 313 litre (69) in male and female patients, respectively. The clearance in both groups was 1.8 litre min-1. Seven out of 12 patients showed significant secondary peaks in blood propofol concentration associated with recovery from anaesthesia.

Adult

Evaluation of an index of peripheral vascular resistance in human subjects.

Based on evidence in animal studies, an index of resistance in man has been evaluated. In 10 human subjects, ranging in age from 31 to 60 years, direct brachial arterial pressure was measured, and a computer program using linear regression calculated the index of resistance from the slope of the logarithm of the pressure versus time during the second half of diastole. Resistance was independently calculated from 20-s averages of pressure and cardiac output obtained by thermodilution. Resistance was varied by infusion of phenylephrine and nitroprusside. The values of the index of resistance were compared with independently calculated resistance by linear regression and correlation. The index of resistance was variably correlated with independently calculated resistance. However, when the comparison was limited to points with a model correlation coefficient greater than or equal to 0.98 (i.e. a good fit of a straight line to the logarithm of pressure versus time), the correlation of the index of resistance with calculated resistance approached or exceeded 0.9 in eight of 10 subjects. In the two subjects showing poor correlation of the index with calculated resistance, estimated compliance from the same model was much lower than in the other subjects. In the same two subjects pressure dependence of estimated compliance was much higher than in the other subjects, suggesting the presence of significant atherosclerosis. We concluded that the diastolic decay of pressure may be used to calculate a useful index of resistance, provided that a single exponential decay fits the observed diastolic waveform well, and arterial compliance is not significantly reduced by atherosclerosis.

Adult

Interaction between penicillin and the DD-carboxypeptidase of the unstable L-form of Proteus mirabilis strain 19.

Binding of penicillin to the DD-carboxypeptidase of the unstable spheroplast L-form of Proteus mirabilis results in the rapid formation of a modified enzyme-inhibitor complex which in turn undergoes rapid decay into reactivated enzyme and an antibiotically inactive penicillin degradation product. Major antibiotic metabolites recovered from such interactions were benzylpenicilloic acid and phenoxymethylpenicilloic acid from benzylpenicillin and phenoxymethylpenicillin, respectively, suggesting a second enzymic function of the DD-carboxypeptidase as a penicillinase of low efficiency. Statistical analyses made with the help of a linear regression program show that the enzyme interacts with the substrate UDP-N-acetylmuramoyl-L-alanyl-D-gamma-glutamyl-(L)-meso-2,6-diaminopimelyl-(L)-D-alanyl-D-alanine and either benzympenicillin or carbenicillin in a non-competitive manner.

Carboxypeptidases

Estimate of the number and of the temporal sequence of measurements in a kinetic experiment.

The optimal choice of the number and the temporal sequence of measurements is that which minimizes the expected error on the parameters of the model later used to fit the observations. Practical constraints regarding the feasibility of a given temporal distribution of measurements, cost-benefit ratio as well as prior knowledge about the parameters and their variability in the selected population must be integrated in the decisional process. A non-linear integer programming procedure is used to find reasonable solutions to this problem.

Models, Biological

Human-like dynamic programming neural networks for dynamic time warping speech recognition.

This paper presents a human-like dynamic programming neural network method for speech recognition using dynamic time warping. The networks are configured, much like human's, such that the minimum states of the network's energy function represent the near-best correlation between test and reference patterns. The dynamics and properties of the neural networks are analytically explained. Simulations for classifying speaker-dependent isolated words, consisting of 0 to 9 and A to Z, show that the method is better than conventional methods. The hardware implementation of this method is also presented.

Humans

Stereotactic radiosurgery for fractionated radiation: a proposal applicable to linear accelerator and proton beam programs.

A stereotactic radiosurgery technique is described which allows stereotactic radiation therapy to be easily fractionated on a daily or weekly basis. This permits adequate and safe radiation therapy to lesions larger than 2.5 cm, such as large arteriovenous malformations, and possibly safer radiation to smaller lesions near crucial intracranial structures. The technique utilizes external scalp landmarks and avoids the need for standard stereotactic head devices.

Equipment Design

Dynamics of Drosophila eye development and temporal requirements of sevenless expression.

The development of the compound eye of Drosophila consists of a linear, stereotyped program starting at the posterior end of the eye imaginal disc and progressing towards the anterior border. The determination of the R7 photoreceptor cells is part of this process and is dependent on the sevenless gene. In this study, we used a heat-shock-inducible sevenless gene as a conditional allele to determine the exact temporal requirements of sevenless gene expression and to reveal the stages of ommatidial development during which the presumptive R7 cell can respond to the presence of sevenless protein. Our results indicate that sevenless gene function is only required during a brief, defined period for the initiation of R7 development; subsequently sevenless is dispensable for both differentiation and function of the R7 photoreceptors. Furthermore, using rescue of R7 cells as an internal marker to monitor the progression of eye development we could examine when and at what rate ommatidial columns form.

Alleles

Comparison of four different algorithms for the calculation of radioimmunoassay standard curves.

Standard curves from 150 determinations of 10 different hormone radioimmunoassays have been evaluated using the following four procedures: logit transformation as performed by Rodbard, Healy's 4-component-logit, spline approximation and a linear interpolation program. The results, calculated by computer, have been compared with manual analysis as the refernce method. The influece of various correction factors has been studied by artificial insertion of outliers. Approximately two thirds of all investigated curves could be calculated with almost equal success using all 4 methods. In calculating the remaining third of the standard curves, each method showed characteristic errors which depended upon the reliability of the assay data and the kind of outliers. The results suggest that the smoothing by spine function is the most flexible method in approximating a radioimmunoassay standard curve. In comparison with the other methods it is less influenced by random errors and fits the characteristic and symmetry of the ideal curve most exactly. The 4-component logit which has been extended by data screening is an improvement of the original logit transformation. Certain reservations exist, however, in elimination of outliers because of the dispersion of the data points, and also because of the arbitrarily set thresholds.

Mathematics

Characterization of opioid binding sites in rat spinal cord.

Binding sites were characterized in rat whole spinal cord crude membrane preparations using selective labelling techniques with multiple methods of mathematical analysis of experimental curves. Mathematical analysis of single [3H]-[D-Ala2,MePhe4,Gly-ol5] enkephalin (DAGO) saturation curves suggested binding of the [3H]-ligand at one site, while displacement curves of low concentrations of [3H]-DAGO with selective mu-ligands indicated the presence of high- and low-affinity sites. All the [3H]-DAGO curves processed simultaneously by LIGAND analysis showed the presence of high (27%) and low (73%) affinity components, with a total Bmax of 3.19 pmol/g tissue. Eighty percent of [3H]-[D-Ala2,D-Leu5] enkephalin (DADLE) binding was displaced by DAGO with high affinity, indicating that a high percentage of [3H]-DADLE binding was at mu-sites. Saturation curves of [3H]-DADLE after inhibition of mu-sites by unlabelled DAGO (delta-sites) were monophasic with non-linear fitting analysis and the Bmax was 0.90 pmol/g tissue. Most mathematical analysis of single saturation curves of [3H]-(-)-bremazocine indicated binding at more than one site. DAGO, DADLE, U-69,593 and PD 117302 displaced 0.15 nM of [3H]-(-)-bremazocine biphasically: the percentages of displacement calculated with the non-linear fitting program were respectively 50 (mu-sites), 64 ((mu + delta)-sites), 18 and 25 (kappa-sites). Haloperidol displaced [3H]-(-)-bremazocine only at microM concentrations. suggesting no binding at sigma-sites. In the presence of 225 nM of DAGO, DADLE displaced only 21% of [3H]-(-)-bremazocine 0.15 nM binding (delta-sites). Most mathematical analysis of saturation curves of [3H]-(-)-bremazocine, after inhibition of binding at mu- and delta-sites by DAGO and DADLE, still indicated binding at more than one site and the selective kappa-ligands U-69,593 and PD 117302 displaced [3H]-(-)-bremazocine in these experimental conditions. LIGAND analysis of saturation and inhibition curves of [3H]-(-)-bremazocine by U-69,593 and PD 117302 showed the presence of high (43%) and low (57%) affinity components, with a total Bmax of 2,73 pmol/g tissue. Thus in rat spinal cord there are at least two mu-sites bound by [3H]-DAGO which amount together to approximately 47% of total opioid sites, delta-sites bound by [3H]-DADLE amounting to approximately 13%, kappa-sites and other unknown sites (possibly a kappa-subtype) bound by [3H]-(-)-bremazocine, which together are approximately 40% of total opioid sites.

Animals

Relationships between linear type scores, objective type measures, and indicators of mastitis.

Objectives were to examine the usefulness of objective measures of conformation and linear scores for type for predicting indicators of mastitis. Approximately 300 cows in five herds were scored in the experimental linear classification program of the Holstein-Friesian Association of America, measured objectively for numerous components of conformation, and evaluated for four indicators of mastitis. Indicators of mastitis were average natural logarithm of somatic cell counts during the lactation, percent of samples (taken approximately every 3 mo) infected during a lactation, percent of samples infected with Staphylococcus aureus, Streptococcus agalactiae, Escherichia coli, Klebsiella, and percent of samples with signs of mastitic milk. Relationships between conformation and mastitis often were inconsistent because of moderate or low correlations between indicators of mastitis. Indicators of mastitis were predicted from results of multiple regression analyses with subjective scores and objective measures of type as independent variables. Largest differences of predicted indicators of mastitis from variation in any single type trait were sufficient to be important economically, but average differences were much smaller. Selection to reduce frequencies of cows with deep udders, especially low rear udders, widely placed teats, rear teats too far back, and teats that are short and wide, may augment, modestly, efforts to reduce incidence of mastitis through improved sanitation, antibiotic treatment, and proper milking procedures.

Animals