A spectrophotometric assay for chlorine-containing compounds.

Determinations of hypochlorous acid and chloramine compounds are important in a number of areas. Several techniques are now available for such analyses, but most require unstable reagents and/or multiple steps in the analytical procedure. We have developed a simple, one-step spectrophotometric assay for reactive chlorine-containing compounds involving the oxidation of ascorbic acid by hypochlorous acid or chloramines. There is no interference from other nonhalide oxidants such as hydrogen peroxide or hypothiocyanous acid. Because small amounts of ascorbic acid will not damage biological materials, this method also allows continuous measurements of the generation of chlorine-containing compounds by activated neutrophils. This simple assay permits precise analysis of as little as 1 nmol of HOCl.     

Excluded volume approximation to protein-solvent interaction. The solvent contact model.

Important properties of globular proteins, such as the stability of its folded state, depend sensitively on interactions with solvent molecules. Existing methods for estimating these interactions, such as the geometrical surface model, are either physically misleading or too time consuming to be applied routinely in energy calculations. As an alternative, we derive here a simple model for the interactions between protein atoms and solvent atoms in the first hydration layer, the solvent contact model, based on the conservation of the total number of atomic contacts, a consequence of the excluded-volume effect. The model has the conceptual advantage that protein-protein contacts and protein-solvent contacts are treated in the same language and the technical advantage that the solvent term becomes a particularly simple function of interatomic distances. The model allows rapid calculation of any physical property that depends only on the number and type of protein-solvent nearest-neighbor contacts. We propose use of the method in the calculation of protein solvation energies, conformational energy calculations, and molecular dynamics simulations.     

Colour-blind drivers' perception of traffic signals.

There is a dangerous and widespread assumption that colour-blind drivers drive as safely as drivers with normal vision. Colour-blind drivers have difficulties recognizing traffic and vehicle signals. However, simple and practical solutions such as changes to the design, colour and shape of signals will aid these drivers.     

Cardamine guatemalensis sp. nov. (Brassicaceae) from Guatemala

Cardamine guatemalensis Al‐Shehbaz is described from Zacapa, Guatemala. Its features such as the perennial, scapose, rhizomatous habit and simple leaves are very rare among North American Brassicaceae. The distinguishing characters from other species of Cardamine are discussed.     

The treatment of Wilson's disease with zinc. IV. Efficacy monitoring using urine and plasma copper

Progress has been made in establishing the efficacy and safety of oral zinc as a maintenance therapy for Wilson's disease. It is important to develop simple, noninvasive monitoring methods to assure the adequacy of zinc therapy in individual patients. In this paper we report the use of 24-hr urine copper and plasma copper measurements to monitor efficacy of zinc maintenance therapy in 30 Wilson's disease patients. In examples of therapeutic inadequacy such as noncompliance, these values increase. With continued long-term adequate therapy, they remain stable or decrease. These two simple monitoring tools appear to be very useful in monitoring Wilson's disease patients receiving zinc therapy.     

A generalizing,analytic model for 2-way cross-classifications. extensions and ramifications. part 1

A generalizing, analytic model is developed for 2-way cross-classifications with fixed class sizes The model is expressed in closed form as a simple operator formula, and is readily extended to n-way cross-classifications, and to such cases where one or more cells are vacuous or fixed. The model permits easy derivation, by means of simple differential operators, of the exact power moments, product moments, and factorial moments of the cell frequencies. From this, the moments of the exact sampling distribution of the conventional x₂-statistic can be computed, which, in turn, leads to a reappraisal of the Chi-square approximation for sparse and isotropic contingency tables. Here, the Gamma distribution is considered, and numerical results are presented that would suggest preference of the Gamma approximation over the Chi-square in such cases.     

The sliding filament model of muscle contraction. I. Quantum mechanical formalism

The quantum mechanical analog of work is defined and discussed by using a simple hypothetical molecular machine, thus enabling the introduction of clearly defined ideas which are necessary for a molecular discussion of biological machines such as the contractile machinery in striated muscle. The problem of control of such quantum machines is discussed and shown to be possible using the concept of a stimulated transition. The problem of “reversibility” is also discussed and shown to have a satisfactory solution for the orders of magnitude of the forces and velocities involved in muscular contractile machinery.     

Simple sequence in brain and nervous system specific proteins.

We examined sequences expressed in the brain and nervous system using EST data. A previous study including sequences thought to have neurological function found a deficiency of simple sequence within such sequences. This was despite many examples of neurodegenerative diseases, such as Huntington disease, which are thought to be caused by expansions of polyglutamine tracts within associated protein sequences. It may be that many of the sequences thought to have neurological function have other additional, non-neurological roles. For this reason, we examined sequences with specific expression in the brain and nervous system, using EST expression data to determine if they too are deficient of simple, repetitive sequences. Indeed, we find this class of sequences to be deficient. Unexpectedly, however, we find sequences expressed in the brain and nervous system to be consistently enriched for histidine-enriched simple sequence. Determining the function of these histidine-rich regions within brain-specific proteins requires more experimental data.     

Studies with Composite Membranes: I. Preparation and Measurement of Impedances

Simple and composite membranes have been prepared from 2% collodion solutions containing different amounts of polystyrenesulfonic acid (PSSA). Various membrane parameters such as water content, electrolyte uptake, exchange capacity, and permselectivity of these membranes have been determined. The resistance and capacitance of simple membranes have been measured as functions of both external electrolyte concentration and internal fixed charge density. The impedance characteristics of composite membranes also have been determined and discussed in terms of the resistance and capacitance characteristics of simple membranes from which the composite structures have been formed.     

Receptor-based assays in screening for biologically active substances.

Molecular biology has identified new receptors and ligands which are deregulated in diseases such as cancer and autoimmune conditions and which provide rational targets for therapeutic intervention. Advances in instrumentation and methodology make it possible to screen large numbers of samples in simple receptor-ligand binding assays in the search for drug candidates. Caution must be exercised in the interpretation of data derived from such assays. This is particularly pertinent to the recently characterized receptors, such as the cytokine receptors, as we do not fully understand the relationship between the receptor type and the linkage of receptors to the appropriate or inappropriate second messenger systems that are used in the experimental screening protocols and the disease state.     

Visual discrimination of simple geometrical patterns: I. Measurements for multiple element stimuli

We describe a method for the measurement of visual discrimination between simple patterns. The target to be discriminated is embedded in a background consisting of multiple, randomly positioned but identical elements, and is distinguished by a single parameter such as magnification or relative rotation. The positions of the target and background elements are varied randomly between presentations and discrimination for different values of the target parameter is measured in terms of the time taken for detection of the target. Using this method, we have studied discrimination of rotation and of magnification for simple pattern elements such as lines, triangles and squares. The results for rotation discrimination are interpreted as evidence for the activity of two discrimination mechanisms, one sensitive to the orientation of the lines from which the pattern elements are constructed and the other to the orientation of the pattern element relative to the visual field.     

Computerisation of diabetic clinic records.

A simple system for putting diabetic records on a computer file is achieved by using stationery that combines the usual handwritten records (not computerised) with the minimum of essential data suitable for punching on to computer tape. The record may be brought up to date at a selected time time interval. This simple, cheap system has been in use in a busy clinic for six years. The information on about 8000 diabetics now held in the computer file is used chiefly to help research by creating registers of patients with specified characteristics, such as treatment, heredity complications, and pregnancy. A complete up-to-date index of the entire clinic population is always available, and routine clinic statistics are returned every six months.     

Multipoint linkage analysis.

A formula is given for the advantage of n-point sampling, which approaches infinity with n. However, 2-point and 3-point analyses extract nearly all the information in such samples and at the same time communicate this information as lods and chi 2, which can be combined with other data by simple addition without reevaluation of the likelihood. When null interference is assumed, map distances and multiple recombination frequencies are inflated, and there is substantial loss of efficiency and of support for the correct order.     

Cell electrophoresis research directed toward clinical cytodiagnosis.

The application of cell electrophoresis to cytodiagnosis requires that a scientifically established basis exists for identifying abnormal cells electrophoretically, that research to detect such differences in the cytodiagnostic setting is possible and that a rapid and simple method of cell electrophoresis is adaptable to the clinical setting. Data are presented indicating modifications of electrophoretic mobility due to herpes simplex virus type 1 infection and Rous sarcoma virus transformation of culture cells. A simple apparatus for electrophoretically separating cells on a density gradient and collecting them for subsequent analysis is described, and results of experiments with this apparatus are consistent with those obtained by microscopic electrophoresis. Laser-doppler spectroscopic electrophoresis is suggested as a rapid method adaptable to clinical application.     

Simple sequence repeats (microsatellites): mutational mechanisms and contributions to bacterial pathogenesis. A meeting review

This review summarises the presentations and discussions that took place during a European Science Foundation-funded workshop whose purpose was to gain current perspectives on the mutational mechanisms of simple sequence repeats and the contribution of localised hypermutation in such repeats to bacterial pathogenesis. In vitro biophysical and biochemical assays of mutational mechanisms were covered as well as genetic studies in various eukaryotic and prokaryotic organisms. Presentations on bacterial pathogenesis elaborated investigations of the use of repeats for typing of strains, epidemiological investigations of mutation rates and functions of loci whose expression is controlled by simple sequence repeats. This review tabulates current perspectives on the cis- and trans-acting factors for mutation of simple sequence repeats and the orientations of mononucleotide repeats in some bacterial species that utilise repeats for adaptation.     

Neighbor joining algorithms for inferring phylogenies via LCA distances.

Reconstructing phylogenetic trees efficiently and accurately from distance estimates is an ongoing challenge in computational biology from both practical and theoretical considerations. We study algorithms which are based on a characterization of edge-weighted trees by distances to LCAs (Least Common Ancestors). This characterization enables a direct application of ultrametric reconstruction techniques to trees which are not necessarily ultrametric. A simple and natural neighbor joining criterion based on this observation is used to provide a family of efficient neighbor-joining algorithms. These algorithms are shown to reconstruct a refinement of the Buneman tree, which implies optimal robustness to noise under criteria defined by Atteson. In this sense, they outperform many popular algorithms such as Saitou and Nei's NJ. One member of this family is used to provide a new simple version of the 3-approximation algorithm for the closest additive metric under the iota (infinity) norm. A byproduct of our work is a novel technique which yields a time optimal O (n (2)) implementation of common clustering algorithms such as UPGMA.     

Cyclic-imide-hydrolyzing activity of D-hydantoinase from Blastobacter sp. strain A17p-4.

The cyclic-imide-hydrolyzing activity of a prokaryotic cyclic-ureide-hydrolyzing enzyme, D-hydantoinase, was investigated. The enzyme hydrolyzed cyclic imides with bulky substituents such as 2-methylsuccinimide, 2-phenylsuccinimide, phthalimide, and 3,4-pyridine dicarboximide to the corresponding half-amides. However, simple cyclic imides without substituents, which are substrates of imidase (ie.g., succinimide, glutarimide, and sulfur-containing cyclic imides such as 2,4-thiazolidinedione and rhodanine), were not hydrolyzed. The combined catalytic actions of bacterial D-hydantoinase and imidase can cover the function of a single mammalian enzyme, dihydropyrimidinase. Prokaryotic D-hydantoinase also catalyzed the dehyrative cyclization of the half-amide phthalamidic acid to the corresponding cyclic imide, phthalimide. The reversible hydrolysis of cyclic imides shown by prokaryotic D-hydantoinase suggested that, in addition to pyrimidine metabolism, it may also function in cyclic-imide metabolism.     

Positive selection for loss of tetracycline resistance.

A simple technique has been devised that allows direct plate selection of tetracycline-sensitive clones from a predominantly tetracycline-resistant population. The technique is especially useful in genetic methodologies based on the use of tetracycline resistance transposons, such as Tn10. Potential uses of the method include selection of deletion mutants, fine-structure mapping, generalized mapping, construction of multiply marked strains, elimination of tetracycline resistance transposons and plasmids and cloning. The technique is based on our finding that tetracycline-resistant cells are hypersensitive to lipophilic chelating agents, such as fusaric acid. This finding supports the contention that certain metal ions critically facilitate tetracycline uptake and leads us to suggest possible molecular mechanisms for tetracycline resistance.     

The lipase activity of a partially purified lipolytic enzyme of Escherichia coli.

Escherichia coli lipase was found to have a broad pH optimum between pH 8 and 10. Long-chain acyl triacylglycerols such as trioleolglycerol were hydrolysed at a relatively slow rate, whereas, the shorter-chain acyl derivative tracapryloylglycerol was not. Triacylglycerols and diacylglycerols were broken down at a rate 10 to 15 fold greater than that for monoacylglycerol. Simple esters such as methyloleate and cetylpalmitate were hydrolysed at rates greater than that of triacyglycerol. Water-soluble esters such as p-nitrophenylacetate were not attacked. Hydrolysis of lipase substrate occurred more readily in the presence of an anionic detergent such as taurocholate. The enzyme had no marked preference for the 1- or 3-position of triacylglycerols but attacked these positions much more readily than position 2. The enzyme also catalyzed transacylation reaction with simple alcohols such as methanol or ethanol.