The influence of creosote compounds on the aerobic degradation of toluene

The inhibiting effect of 14 typical creosote compounds on the aerobic degradation of toluene was studied in batch experiments. Four NSO-compounds (pyrrole, 1-methylpyrrole, thiophene, and benzofuran) strongly inhibited the degradation of toluene. When the NSO-compounds were present together with toluene, little or no degradation of toluene was observed during 16 days of incubation, compared with a total removal of toluene within 4 days when the four compounds were absent. Indole (an N-compound) and three phenolic compounds (phenol, o-cresol, and 2,4-dimethylphenol) also inhibited the degradation of toluene, though the effect was much weaker that of the four NSO-compounds. O-xylene, p-xylene, naphthalene and 1-methylnaphthalene seemed to stimulate the degradation even though the influence was very weak. No effects of benzothiophene (an S-compound) and quinoline (an N-compound) were observed. Benzofuran (an O-compound) was identified as the compound that most inhibited the degradation of toluene. An effect could be detected even at low concentrations (40 g/l).Abbreviations bf benzofuran - bt benzothiophene - dmp 2,4-dimethylphenol - GC gas chromatograph - ind indole - mnap 1-methylnaphthalene - MAH monoaromatic hydrocarbons - mpyr 1-methylpyrrole - nap naphthalene - o-cre o-cresol - o-xyl o-xylene - phe phenol - pyr pyrrole - p-xyl p-xylene - tol toluene - thi thiophene - qui quinoline     

Prediction of location of active sites in biologically active peptides

In a previous paper we demonstrated that the short-range compact regions in atrial natriuretic factor (-hANF) predicted by the average distance map (ADM) correspond to its active sites [Kikuchi,J. Protein Chem.11, 579–581 (1992)]. In the present paper we apply the same method to other bioactive peptides and peptidic enzyme inhibitors. We again observe that active sites in each peptide are contained in short-range compact regions predicted by the ADM for the peptide. This demonstrates that the ADM method predicts the possible location of active sites in biologically active peptides in general. The possibility of practical application of the present method to rational drug design is also discussed.     

Hume and the argument for biological design

There seems to be a widespread conviction — evidenced, for example, in the work of Mackie, Dawkins and Sober — that it is Darwinian rather than Humean considerations which deal the fatal logical blow to arguments for intelligent design. I argue that this conviction cannot be well-founded. If there are current logically decisive objections to design arguments, they must be Humean — for Darwinian considerations count not at all against design arguments based upon apparent cosmological fine-tuning. I argue, further, that there are good Humean reasons for atheists and agnostics to resist the suggestion that apparent design — apparent biological design and/or apparent cosmological fine-tuning — establishes (or even strongly supports) the hypothesis of intelligent design.     

Development of a serum-free medium using computer-assisted factorial design and analysis

Stepwise screening of media supplements using factorial design and analysis was employed in the development of serum-free medium for a recombinant Chinese hamster ovary cell line. The effects (growth and target protein production) of different combinations were measured at two time points to ensure adequate response. The results were analysed by a computer program specialized in factorial analysis. The formulation deduced from the previous experiment was used as the new basal medium for the next screening. Certain significant nutrients were studied again in a more advanced formulation in order to analyse the potential synergistic effects with new media components. Compared to cells grown in serum-containing medium, cells adapted to the final formulation of the serum-free medium had a comparable growth rate but a four fold increase in the active protein production.Abbreviations ANOVA Analysis of variance - BSA bovine serum albumin - CHO Chinese hamster ovary - FBS fetal bovine serum - MTT 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide - PBS phosphate buffered saline - SFM serum-free medium     

The Western Rock lobster Panulirus cygnus (George, 1962) (Decapoda: Palinuridae): the effect of temperature and developmental stage on energy requirements of pueruli

Pueruli of the Western Rock Lobster Panulirus cygnus (George, 1962) are thought to be nonfeeding. Consequently, the metabolic rate is expected to be low during this stage in order to conserve energy reserves. Furthermore, since water temperature potentially has a substantial impact on energetic needs, the puerulus possibly exhibits mechanisms to reduce the effect of temperature on energy consumption. To test these propositions the metabolic rate was measured in post-settlement pueruli and in juveniles at two water temperatures. A respirometer of variable volume (10–50 ml) was designed for this purpose, incorporating a dark-type oxygen sensor. Results were compared with data available from the literature.

Oxygen consumption in pre-molt pueruli and in intermolt juveniles (1.48 to 5.65 μmol O₂· individual⁻¹·h⁻¹ on average) was substantially higher than in post-settlement pueruli (1.06–1.41μmol O₂·ind.⁻¹h⁻¹). These significant changes could only partly be explained through changes in biomass. Furthermore, no significant effect of an increased water temperature (from 18 to 23 ° C) could be detected on the metabolic rate in post-settlement pueruli, and the effect is moderate in pre-molt pueruli (Q₁₀ = 1.95). The water temperature has, however, a substantially greater impact on first and second molt juveniles (Q₁₀ = 2.46 to 4.80).

The energetic demand was calculated from oxygen consumption and indicate that energetic needs of post-settlement pueruli is low compared with both pre-molt pueruli and juveniles. A low energetic demand and a reduced effect of temperature on energy consumption is of considerable benefit to a non-feeding larva, and may provide the puerulus with a means of extending the duration of the non-feeding stage and increasing the chance for survival beyond metamorphosis to the first feeding stage. Results indicate that the energetic demand during metamorphosis may be considerable. It is postulated that energetic requirements of the planktonic (actively swimming) puerulus larvae are considerably higher and are likely to be more temperature dependent.     

Predicting mean and variance of all possible lines and hybrids from designs with partially inbred progenies

Two-factor mating designs at consecutive S_n and S_(n+1) levels (S₀ and S₁ S₁ and S₂, or F₂ and F₃) allow estimation of all components of the variation among homozygous lines and F₁ hybrids that can be derived from a given population. They also allow for the prediction of the mean of these lines and single-cross hybrids. Some tests for the presence of epistasis are possible at the levels of means and of variances. Such mating designs can be very useful for predicting the value of the best possible lines or the best possible F₁ hybrids when it is difficult to produce, at an experimental level for exploratory purposes, either lines or hybrids.     

Rational design of peptide-based inhibitors of trypanothione reductase as potential antitrypanosomal drugs

Summary The rational design of ligands for the substrate-binding site of a homology-modelled trypanothione reductase (TR) was performed. Peptides were designed to be selective for TR over human glutathione reductase (GR). The design process capitalized on the proposed differences between the activesites of TR and human GR, subsequently confirmed by the TR crystal structure. Enzyme kinetics confirmed that forT. cruzi TR benzoyl-Leu-Arg-Arg-ß-naphthylamide was an inhibitor (K_i 13.8µM) linearly competitive with the native substrate, trypanothione disulphide, and did not inhibit glutathione reductase.     

The making of the minibody: An engineered β-protein for the display of conformationally constrained peptides

Conformationally constraining selectable peptides onto a suitable scaffold that enables their conformation to be predicted or readily determined by experimental techniques would considerably boost drug discovery process by reducing the gap between the discovery of a peptide lead and the design of a peptidomimetic with a more desirable pharmacological profile. With this in mind, we designed the minibody, a 61-residue β-protein aimed at retaining some desirable features of immunogloblin variable domains, such as tolerance to sequence variability in selected regions of the protein and predictability of main chain conformation of the same regions, based on the ‘canonical structures’ model. To test the ability of the minibody scaffold to support functional sites we also designed a metal binding version of the protein by suitably choosing the sequences of its loops. The minibody was produced both by chemical syntyhesis and expression in E. coli and charactgerized by size exclusion chromatography, UV CD (circular dichroism) spectroscopy and metal binding activity. All our data supported the model, but a more detailed structural characterization of the molecule was impaired by its low soubility. We were able to overcome this problem both by further; mutagenesis of the framework and by addition of a solublizing motif. The minibody is being used to select constrained human IL-6 peptidic ligands from a library displayed on the surface of the f1 bacteriophage.