A differential scanning calorimetry study of the thermal denaturation of Bacillus thermoproteolyticus rokko thermolysin was carried out. The calorimetric traces were found to be irreversible and highly scan-rate dependent. The shape of the thermograms, as well as their scan-rate dependence, can be explained by assuming that the thermal denaturation takes place according to the kinetic scheme N k----D, where k is a first-order kinetic constant that changes with temperature, as given by the Arrhenius equation, N the native state, and D the unfolded state or, more probably, a final state, irreversibly arrived at from the unfolded one. On the basis of this model, the value of the rate constant as a function of temperature and the activation energy have been calculated. It is shown that the proposed model may be considered as being one particular case of that proposed by Lumry and Eyring [Lumry, R., & Eyring, H. (1954) J. Phys. Chem. 58, 110] N in equilibrium D----I, where N is the native state, D the unfolded one, and I a final state, irreversibly arrived at from D. Lastly, some comments are made on the use of the scan-rate effect on the calorimetric traces as an equilibrium criterion in differential scanning calorimetry.

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Flavodoxins are ar/p proteins that mediate electron transfer reactions. The conformational stability of apoflavodoxin from A n a b~n a PCC 71 19 has been studied by calorimetry and urea denaturation as a function of pH and ionic strength. At pH > 12, the protein is unfolded. Between pH 1 1 and pH 6, the apoprotein is folded properly as judged from near-ultraviolet (UV) circular dichroism (CD) and high-field 'H NMR spectra. In this pH interval, apoflavodoxin is a monomer and its unfolding by urea or temperature follows a simple two-state mechanism. The specific heat capacity of unfolding for this native conformation is unusually low. Near its isoelectric point (3.9), the protein is highly insoluble. At lower pH values (pH 3.5-2.0), apoflavodoxin adopts a conformation with the properties of a molten globule. Although apoflavodoxin at pH 2 unfolds cooperatively with urea in a reversible fashion and the fluorescence and far-UV CD unfolding curves coincide, the transition midpoint depends on the concentration of protein, ruling out a simple two-state process at acidic pH. Apoflavodoxin constitutes a prornising system for the analysis of the stability and folding of cr/0 proteins and for the study of the interaction between apoflavoproteins and their corresponding redox cofactors.

Hyperpolarised gases have been most recently used in magnetic resonance imaging to demonstrate new image-derived pulmonary function parameters. One of these parameters is the apparent diffusion coefficient, which reflects the sizes of the structures that compartmentalise gas within the lung (i.e. alveolar space). In the present study, noninvasive parameters were compared to microscopic measurements (mean linear intercept and mean alveolar internal area).Nonselective helium-3 gas density coronal ex vivo images and apparent diffusion maps were acquired in control and elastase-induced panacinar emphysema rats. Total lung capacity was considered the reference for both imaging experiments and lung fixation.A mild degree of emphysema was found based on mean linear intercept (134¡25 mm) versus control (85¡14 mm). The apparent diffusion coefficients were significantly different between the two groups (0.18¡0.02 and 0.15¡0.01 cm 2 ?s -1 for elastase and control, respectively).A significant correlation between the apparent diffusion coefficient and corresponding morphometric parameters in mild emphysema was demonstrated for the first time. This study opens the possibility of estimating absolute airspace size using noninvasive techniques.

a b s t r a c tThe role of iron on the degradation of different organic compounds, differing in their structure (aliphatic versus aromatic) and iron complex formation capacity, by conventional and photo-Fenton processes was investigated. Results show that these chemical characteristics can affect the degree of treatment in terms of COD and TOC removals. While aromatics exhibited a fast and great reduction in the COD by the conventional Fenton process, aliphatic compounds, apart from acetic acid, required the presence of UV light to enhance treatment results. EDTA and oxalic acid responded very positively to UV irradiation in both COD removal and mineralization, reaching the highest values showed by aromatics; and results depended on the intensity of the UV light applied. Phenol and 4-nitrophenol responded favourably to UV irradiation in terms of mineralization and slightly in COD removal. Reductions in the COD were almost total (95.99%), while only an 80% of reduction in the TOC was achieved, for the best photo-Fenton treatment of oxalic acid, phenol and nitrophenol. 60% COD and 40% TOC removals were achieved correspondingly in the case of EDTA. Acetic acid showed almost no mineralization and low COD removal (≈20%) when treated by a conventional Fenton process; and did not enhanced results when assisting the treatment with UV light. Photo-regeneration of ferrous ion and photo-decarboxylation of iron carboxylates are assessed in the framework of these results.