Several beta-lactamases, enzymes that play an important part in antibiotic resistance, have been purified by affinity chromatography on boronic acid gels. The procedure is rapid, appears to be selective for beta-lactamases, and allows a one-step purification of large amounts of enzyme from crude cell extracts. We have found the method useful for any beta-lactamase that is inhibited by boronic acids. Two kinds of boronic acid column have been prepared, the more hydrophobic one being reserved for those beta-lactamases that bind boronic acids relatively weakly. beta-Lactamase I from Bacillus cereus, P99 beta-lactamase and K 1 beta-lactamase from Gram-negative bacteria are among the better-known beta-lactamases that have been purified by this method. The procedure has also been used to purify a novel beta-lactamase from Pseudomonas maltophilia in high yield; the enzyme has an exceptionally broad substrate profile and hydrolyses monocyclic beta-lactams such as azthreonam and desthiobenzylpenicillin.
Evidence will be presented which demonstrates that environment sensitive solvatochromic dyes can be used to detect the presence of the full range of homeopathic potencies through changes in their visible spectra. These dyes are characterised by possessing an oscillating dipole and it will be shown that this feature is essential for their interaction with potencies by comparison with structurally comparable non-solvatochromic dyes, which show no evidence of any sensitivity to homeopathic medicines.
It will be shown that changes in the spectra of solvatochromic dyes are due to the way these dyes aggregate, or order, in solution as a consequence of their interaction with potencies. Certain solvatochromic dyes appear to have their level of ordering enhanced by potencies, whereas others have their ordering diminished by potencies.
A hypothesis which explains the results presented will be offered and suggests that homeopathic potencies themselves may be oscillating dipoles.
Implications for clinical practice and possible connections between the relevant dye chemistry and some clinically observed effects of homeopathic medicines will be explored in the light of the discoveries being made using these fascinating and informative dyes.
The chemistry involved will be kept as simple as possible for those with a limited chemistry background, whilst maintaining the level of scientific detail necessary for an understanding of the results presented.
A possible explanation for these observations is offered in terms of a potency - induced electron density shift in BDF. The interaction of homeopathic potencies with solvatochromic dyes, particularly BDF, points towards a possible physico-chemical model for the nature of potencies, how they may be interacting with this class of dyes, and moreover how their biological effects may be mediated.
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