The effect of antibiotics on the photocycle and protoncycle of purple membrane suspensions

The interrelation was studied between the phototransient absorbing maximally at 412 nm (M₄₁₂) and light-induced proton release under steady-state conditions in aqueous suspensions of ‘purple membrane’ derived from Halobacterium halobium. The decay of M₄₁₂ was slowed down by the simultaneous application of the ionophoric antibiotics valinomycin and beauvericin. The former had only slight activity alone and the latter was effective only in conjunction with valinomycin. The steady-state concentration of M₄₁₂ which was formed on illumination was a direct function of the concentration of valinomycin. Maximum stabilization of M₄₁₂ was obtained when the valinomycin was approximately equimolar with the bacteriorhodopsin. Addition of salts to the medium increased the number of protons released per molecule of M₄₁₂ without affecting the level of M₄₁₂ which was produced by continuous illumination. The effectiveness of the salts in this respect depended on the nature of the cation. Ca²⁺ and their antagonists La³⁺ and ruthenium red were found to have especially high affinity for the system. The extent of light-induced acidification could not be enhanced by increasing the pH of the medium from 6.5 to 7.8. The possible mechanism of action of the ionophores and of the cations on the photocycle and on the proton cycle is discussed.     

X-ray absorption spectroscopy of selenium-containing amino acids

Received: 2 April 1999 / Accepted: 17 September 1999     

Computational Oral Absorption Simulation for Low‐Solubility Compounds

Bile micelles play an important role in oral absorption of low‐solubility compounds. Bile micelles can affect solubility, dissolution rate, and permeability. For the pH–solubility profile in bile micelles, the Henderson–Hasselbalch equation should be modified to take bile‐micelle partition into account. For the dissolution rate, in the Nernst–Brunner equation, the effective diffusion coefficient in bile‐micelle media should be used instead of the monomer diffusion coefficient. The diffusion coefficient of bile micelles is 8‐ to 18‐fold smaller than that of monomer molecules. For permeability, the effective diffusion coefficient in the unstirred water layer adjacent to the epithelial membrane, and the free fraction at the epithelial membrane surface should be taken into account. The importance of these aspects is demonstrated here using several in vivo and clinical oral‐absorption data of low‐solubility model compounds. Using the theoretical equations, the food effect on oral absorption is further discussed.     

In vitro dynamics of chromatin organization and migration

The organization of eukaryotic chromatin is not static but changes as a function of cell status during processes such as proliferation, differentiation, and migration. DNA quantification has not been used extensively to investigate chromatin dynamics in combination with cellular migration. In this context, an optimized DNA-specific, nonperturbant method has been developed for studying chromatin organization, using the fluorescent vital bisbenzimidazole probe Hoechst 33342: this property has been described by Hamori et al. (1980). Computer-assisted image analysis was used to follow migratory activity and chromatin organization of L929 fibroblasts during in vitro wound healing. Cell movements were analyzed using an optical flow technique, which consists in the calculation of the velocity field of cells and nuclear movements in the frame. This system allows the correlation of cell migration and position in the cell cycle. It makes it possible to study chromatin dynamics using a quantitative analysis of nuclear differentiation reorganization (nuclear texture) and to correlate this with migration characteristics. The present system would be of interest for studying cell-extracellular matrix interactions using differing substrates, and also the migratory response to chemotactic factors. Such a model is a prerequisite for gaining better understanding of drug action.     

Role of Activity of Gastrointestinal Microflora in Absorption of Calcium and Magnesium in Rats Fed β1-4 Linked Galactooligosaccharides

Rats fed a diet containing β1-4 linked galactooligosaccharides (GOS) (5 g/100 g of diet) absorbed calcium and magnesium more efficiently than those fed the control diet. However, the increment obtained through GOS-feeding was reduced by neomycin sulfate 0.67 g/100 g of diet). Since the decrease in cecal pH in rats fed GOS was suppressed by neomycin-feeding, bacterial action in the digestive tract was considered to be reduced by neomycin-feeding. Our findings suggest that the action of intestinal bacteria is necessary for the effects of GOS.     

Inter-chromophore interactions in pigment-modified and dimer-less bacterial photosynthetic reaction centers

The magnitudes of inter-chromophore interactions in bacterial photosynthetic reaction centers are investigated by measuring absorption and Stark spectra of reaction centers in which monomeric chromophores are modified and in a novel triplet mutant which lacks the special pair. The circular dichroism spectrum of the triple mutant reaction center was also measured. Only small changes in the spectroscopic properties are observed, as has also been found for several types of reaction centers in which the absorption or chemical properties of a chromophore are altered by site-specific mutations. We conclude that the electronic absorption, circular dichroism and Stark features of the special pair and the monomeric chromophores in the reaction center are relatively insensitive to inter-chromophore interactions.     

PHOTOACCLIMATION IN THE TROPICAL CORALLINE ALGA HYDROLITHON ONKODES (RHODOPHYTA, CORALLINACEAE) FROM A FRENCH POLYNESIAN REEF

Pigment concentration, in vivo absorption, and photosynthetic parameters of the coralline alga Hydrolithon onkodes (Heydrich) Penrose and Woelkerling were compared among samples from a lagoon and from a reef crest of Tahiti Island. Four groups of specimens were considered, differing in their natural exposure to PAR. For specimens collected from the lagoon, the tissues from low-light samples had significantly higher pigment concentration, particularly chl a and phycobilins, compared with the high-light exposed plants that contained more total carotenoids. The in vivo absorption spectra normalized to chl a (called a* values) also revealed differences. The low-light samples had a reduced absorption capacity and a well-marked phycobilin absorption signature, whereas sunlit samples showed a greater absorption at wavelengths absorbed mainly by chl a and carotenoids. The decrease of a* when pigment concentration increased is interpreted as a consequence of the pigment packaging. Significantly lower α (chlorophyll basis) and higher E_k values were found in the shaded plants. The values of P _max for the four groups of specimens were not significantly different. The samples showed various degrees of photoinhibition depending on the light exposure during growth, and this effect was more pronounced in the shaded plants. The specimens from the reef crest deviate from the general model presented for the lagoon samples and show a mix of sun- and shade-exposed characteristics. We have shown that the coralline alga H. onkodes responds to its light environment, probably by acclimation rather than ecotypic genetic variation, by adjusting its physiology, but some morphological differences are also involved. Photoacclimation can explain partly the wide distribution of this species over the reef ecosystem and its major contribution to the building of the reef.     

Survey of chemical speciation of trace elements using synchrotron radiation

Information concerning the chemical state of trace elements in biological systems generally has not been available. Such information for toxic elements and metals in metalloproteins could prove extremely valuable in the elucidation of their metabolism and other biological processes. The shielding of core electrons by binding electrons affect the energy required for creating inner-shell holes. Furthermore, the molecular binding and symmetry of the local environment of an atom affect the absorption spectrum in the neighborhood of the absorption edge. X-ray absorption near-edge structure (XANES) using synchrotron radiation excitation can be used to provide chemical speciation information for trace elements at concentrations as low as 10 ppm. The structure and position of the absorption curve in the region of an edge can yield vital data about the local structure and oxidation state of the trace element in question. Data are most easily interpreted by comparing the observed edge structure and position with those of model compounds of the element covering the entire range of possible oxidation states. Examples of such analyses will be reviewed.