Analysing the evacuation procedures employed on a Thames passenger boat using the maritimeEXODUS evacuation model

On the 19 June 2001, a Thames passenger/tour boat underwent several evacuation trials. This work was conducted in order to collect data for the validation of marine-based computer models. The trials involved 111 participants who were distributed throughout the vessel. The boat had two decks and two points of exit from the lower deck placed on either side of the craft, forward and aft. The boat had a twin set of staircases towards the rear of the craft, just forward of the rear exits. maritimeEXODUS was used to simulate the full-scale evacuation trials conducted. The simulation times generated were compared against the original results and categorised according to the exit point availability. The predictions closely approximate the original results, differing by an average of 6.6% across the comparisons, with numerous qualitative similarities between the predictions and experimental results. The maritimeEXODUS evacuation model was then used to examine the evacuation procedure currently employed on the vessel. This was found to have potential to produce long evacuation times. maritimeEXODUS was used to suggest modifications to the mustering procedures. These theoretical results suggest that it is possible to significantly reduce evacuation times.

Structure and denaturation of 4-chlorobenzoyl coenzyme A dehalogenase from Arthrobacter sp. strain TM-1

The secondary structure of the trimeric protein 4-chlorobenzoyl coenzyme A dehalogenase from Arthrobacter sp. strain TM-1, the second of three enzymes involved in the dechlorination of 4-chlorobenzoate to form 4-hydroxybenzoate, has been examined. E(mM) for the enzyme was 12.59. Analysis by circular dichroism spectrometry in the far uv indicated that 4-chlorobenzoyl coenzyme A dehalogenase was composed mostly of alpha-helix (56%) with lesser amounts of random coil (21%), beta-turn (13%) and beta-sheet (9%). These data are in close agreement with a computational prediction of secondary structure from the primary amino acid sequence, which indicated 55.8% alpha-helix, 33.7% random coil and 10.5% beta-sheet; the enzyme is, therefore, similar to the 4-chlorobenzoyl coenzyme A dehalogenase from Pseudomonas sp. CBS-3. The three-dimensional structure, including that of the presumed active site, predicted by computational analysis, is also closely similar to that of the Pseudomonas dehalogenase. Study of the stability and physicochemical properties revealed that at room temperature, the enzyme was stable for 24 h but was completely inactivated by heating to 60 degrees C for 5 min; thereafter by cooling at 1 degrees C min(-1) to 45 degrees C, 20.6% of the activity could be recovered. Mildly acidic (pH 5.2) or alkaline (pH 10.1) conditions caused complete inactivation, but activity was fully recovered on returning the enzyme to pH 7.4. Circular dichroism studies also indicated that secondary structure was little altered by heating to 60 degrees C, or by changing the pH from 7.4 to 6.0 or 9.2. Complete, irreversible destruction of, and maximal decrease in the fluorescence yield of the protein at 330-350 nm were brought about by 4.5 M urea or 1.1 M guanidinium chloride. Evidence was obtained to support the hypothetical three-dimensional model, that residues W140 and W167 are buried in a non-polar environment, whereas W182 appears at or close to the surface of the protein. At least one of the enzymes of the dehalogenase system (the combined 4-chlorobenzoate:CoA ligase, the dehalogenase and 4-hydroxybenzoyl coenzyme A thioesterase) appears to be capable of association with the cell membrane.