Direct determination of endoglucanase activity on cellulose insoluble fibres

Summary A procedure is proposed for the measurement of endoglucanase activity on insoluble cellulose. It is based on the Nelson-Somogyi technique and its relative merits as opposed to a viscometric method are detailed. A relevant characteristic of this procedure concerns its ability to determine the degree of polymerisation of cellulose insoluble fibres.     

A simple assay for methionine adenosyltransferase using cation exchange paper and liquid scintillation spectrometry

A rapid, sensitive and simple procedure for the assay of l-methionine-S-adenosyltransferase based on the use of phosphocellulose ion-exchange paper is presented. The analytical procedure may be generally useful for all enzymes where there is a large cationic charge difference between the substrates and products. An application of a simpllified counting procedure for radioisotopes on solid supports is presented.     

The use of the maharanobis and modified distances for the improvement of simulation of glutamic acid production

A modified simulation procedure based on a statistical approach was investigated. The procedure predicts the time course of fed-batch culture for glutamic acid production by a temperature-sensitive strain of Brevibacterium flavum. The statistical approach requires only a data base of state points obtained in experiments, and not perfect identification of fermentation models. The simulation procedure is based on regression analysis to estimate specific rate parameters of system equations using the data points selected with reference to the Euclid distance. It was modified in that the data selection procedure included the use of the Maharanobis distance as well as a modified distance defined in this study. Simulation results using the modified procedure allow reasonable prediction of the time course of fed-batch culture for glutamic acid compared to that involving the Euclid distance alone.     

Diagnostic evidence for the presence of beta-agonists using two consecutive derivatization procedures and gas chromatography-mass spectrometric analysis

A GC-MS procedure for the detection of different beta-agonists in urine samples based on two consecutive derivatization steps is described. The derivatization procedure is based on the consecutive formation of cyclic methylboronate derivatives followed by a second derivatization step with MSTFA on the same extract, forming TMS derivatives. Injections in the GC-MS system may be carried out after each one of the derivatization steps, obtaining enough information for unambiguous identification. Limits of detection for the two derivatization steps ranged from 0.5 to 5 ng/ml. This procedure was tested with the beta-agonists bambuterol, clenbuterol, fenoterol, formoterol, salbutamol, salmeterol, alpha-hydroxy-salmeterol and terbutaline.     

The determination of glucosyltransferase

Summary A new procedure for the determination of glucosyltransferase (EC 2.4.1.24) activity in amyloglucosidase preparations has been developed. The procedure is based on the rapid separation of glucosyltransferase from amyloglucosidase on carboxymethyl bio-gel A.     

Enzyme assay of L-myo-inositol 1-phosphate synthetase based on high-performance liquid chromatography of benzoylated inositol

A procedure for the determination of inositol by reversed-phase HPLC is described which is based on a precolumn benzoylation and detection at 230 nm. This procedure was used to assay the activity of L-myo-inositol 1-phosphate synthetase (EC 5.5.1.4) after treatment of the enzymatic product by a phosphatase.     

A two-stage procedure for bioequivalence studies

A two-stage experimental procedure for bioequivalence studies is proposed. The procedure is based on the idea that information from a first-stage experiment can be used to form a predictive distribution for the outcome of a second-stage experiment, thus permitting an assessment of the probability of a successful overall outcome. A systematic numerical study of a variety of possible strategies results in the identification of procedures that lead to substantial increases in efficiency compared with single-stage studies.     

Determination of the equilibrium constants of self-associating protein systems. X. Evaluation of the weight fraction of monomer from the diffusion coefficient as a function of concentration

A simplified procedure for determining the mode of association and weight fraction of monomer of a self-associating protein system based on the average diffusion coefficient as a function of concentration is described. This procedure is applicable to cases of monomer-n-mer, three species or indefinite association.     

Purification of antigen B from Taenia solium cysticerci by affinity to mammalian collagen

A 1-step procedure for the easy and rapid purification of milligram amounts of antigen B from a crude extract of Taenia solium cysticerci is described here. This procedure takes advantage of the property of the antigen B to bind to collagen and is based on antigen adsorption to polymeric collagen.     

A test for a threshold in an ordered sequence of correlated proportions

A test procedure using chi-square statistics is proposed for determining a threshold in an ordered sequence of correlated proportions. The procedure is based on the multivariate Bernoulli model. It is applied to the problem of ascertaining when visual acuity has stabilized in a group of patients with regular follow-up after a vision-reducing acute abnormality.     

Adsorption and elution characteristics of nucleic acids on silica surfaces and their use in designing a miniaturized purification unit

We report nucleic acid (NA) adsorption isotherms and elution profiles for silica surfaces and use these to design a miniaturized NA purification unit based on solid-phase extraction with silica beads. The procedure is based on a pressure drop equation for flow through a packed bed and allows estimation of key design parameters such as channel dimensions, liquid flow rates, sample volume, and amount of silica needed. The usefulness of this design procedure is demonstrated by applying it to a column-based NA purification device for influenza detection for a case study of Madin-Darby canine kidney cells infected with influenza A virus.     

Regime analysis and scale-down: Tools to investigate the performance of bioreactors

Scale-up and optimization of biotechnological processes on a large scale tend to be more methodically approached than the application of rules of thumb, experience, and trial and error. Methods frequently used in chemical engineering are adopted in biochemical engineering and are employed with great effect. A summary is given of methods and rules of thumb used in scaling up chemical processes. A procedure to scale up and optimize bioreactors is presented. It is based on the so-called scale-down approach. Some elements of this procedure, viz. theoretical regime analysis and small-scale investigations, are extensively demonstrated by examples. It is shown that a regime analysis based on characteristic times can give a quick estimation of the performance of bioreactors. Small-scale experiments based on the result of such analysis or on the results of a dimensional analysis can give valuable information for scale-up or optimization fermentation processes.     

The determination of folylpolyglutamate synthetase.

A relatively rapid radiochemical procedure for the determination of folylpolyglutamate synthetase activity is presented in this communication. The procedure is based on measurement of the incorporation of radioactive l-glutamate into tetrahydropteroylpolyglutamate on incubation with a tetrahydrofolate. After deproteinating the incubation mixtures with trichloroacetic acid, folate is separated from radioactive glutamate by an adaptation of a procedure generally employed in the isolation of folate from natural materials, i.e., adsorption on columns of charcoal from which it is subsequently eluted with aqueous-alcoholic ammonia containing mercaptoethanol and counted. The procedure is applicable to monitoring purification of the enzymes and to the study of their properties.The technique for separating a radioactive product of enzyme action from a radioactive precursor with a column of charcoal, that has been developed for this procedure is applicable also to other radiochemical enzyme determinations requiring the separation of an aromatic from an aliphatic metabolite.     

Electrostatic potentials in membrane systems

A membrane with an arbitrary distribution of fixed charges inside and on its surfaces is considered. A procedure for calculating the local electrostatic potential at an arbitrary point of the system is described and its validity discussed. This procedure is based on the linearization of the 3-dimensional Poisson-Boltzmann equation around an exact 1-dimensional solution.     

A runs test based on run lengths

A procedure is proposed for testing the hypothesis that Bernoulli trials (successes and failures) are independent with common probability of success. Equivalently, the procedure may be used to test the hypothesis that the arrangement of a fixed number of successes and failures was determined randomly. The procedure is based on a weighted linear combination of the variances of run lengths of successes and failures. It is shown to have desirable asymptotic properties and to be generally more powerful than the usual runs test, while preserving computational simplicity.     

Measures of Imbalance for Unbalanced Models

In this paper we present a procedure to measure the degree of imbalance of an unbalanced data set. The procedure is based on choosing an appropriate loglinear model for the subclass frequencies of the data. A measure of imbalance is then introduced as some function of the chi-squared statistic used in the goodness-of-fit test for the loglinear model. The proposed procedure can also be used to measure departures from certain types of balance, such as proportionality of subclass frequencies, partial balance, and last-stage uniformity.     

A class of goodness of fit tests for a copula based on bivariate right-censored data

The copula of a bivariate distribution, constructed by making marginal transformations of each component, captures all the information in the bivariate distribution about the dependence between two variables. For frailty models for bivariate data the choice of a family of distributions for the random frailty corresponds to the choice of a parametric family for the copula. A class of tests of the hypothesis that the copula is in a given parametric family, with unspecified association parameter, based on bivariate right censored data is proposed. These tests are based on first making marginal Kaplan-Meier transformations of the data and then comparing a non-parametric estimate of the copula to an estimate based on the assumed family of models. A number of options are available for choosing the scale and the distance measure for this comparison. Significance levels of the test are found by a modified bootstrap procedure. The procedure is used to check the appropriateness of a gamma or a positive stable frailty model in a set of survival data on Danish twins.     

Optimal foraging in patches: A case for stochasticity

Like much mathematical modeling in biology, most optimal foraging theory is developed from deterministic analogs of basically stochastic processes. Unlike other models, however, it cannot depend on laws of large numbers to justify this simplification; ignoring stochasticity can lead to wrong answers. This is demonstrated for a predator searching spatially separated patches of prey; it is shown that the choice of an optimal procedure for deciding when to leave a patch must be based on a stochastic model—a predator whose procedure is based on a deterministic model can do arbitrarily badly by comparison with the stochastic optimizer. A general solution is given, and its complexity suggests some objections to standard optimality arguments, and some possible alternatives.     

Determination of urease activity by thermal conductivity gas chromatography

A rapid, simple and inexpensive procedure for the determination of urease activity by using a thermal conductivity gas chromatography method is presented. The procedure is based on the determination of CO2 released in the urease reaction, and showed low coefficient of variation (c.v. less than 1%) and high sensitivity (detection limit 10(-12) mol). This procedure is also suitable for determination of other decarboxylating enzyme activities.     

Rapid radioimmunoassay for guanosine 3',5'-cyclic monophosphate using tritiated ligand.

A radioimmunoassay procedure for guanosine 3',5'-cyclic monophosphate (CGMP) is described. The procedure is based on competitive binding between [3H]CGMP and non-radioactive CGMP, with separation of bound and unbound CGMP by Millipore filtration. The binding reaction showed very high specificity to CGMP, had a broad pH optimum, and reached equilibrium within a short time. A simple procedure for the pruification of assay samples using Dowex AG 50W-X2 resin is also described. CGMP contents in urine samples were assayed without purification. Injection of glucagon into healthy human volunteers resulted in a small but significant reduction in urinary CGMP level, whereas CAMP excretion increased dramatically.