Pheophytin-mediated energy storage of photosystem II particles detected by photoacoustic spectroscopy

The photoacoustic (PA) characteristics (energy storage and heat dissipation) of photosystem II (PSII) core-enriched particles from barley were studied (i) in conditions where there was electron flow, i.e., in the presence of a combination of the electron acceptor K₃ Fe (CN)₆, referred to as FeCN, and the electron donor diphenylcarbazide (DPC), and (ii) in conditions where electron flow was suppressed, i.e., in the absence of FeCN and DPC. The experimental data show that a decrease of heat dissipation with a minimum at 540 nm can be interpreted as energy storage resulting from the presence of pheophytin (Pheo) in the PSII particles. On account of the capability of the PA method to measure the energy absorbed by the chromophores which is converted to heat, it is suggested that the PA detection of Pheo present in the PSII complex will permit to clarify the function of processes involving non-radiative relaxation of excited states in P680-Pheo-Q_A interactions.Abbreviations -Car -Carotene - Chl Chlorophyll - DPC Diphenylcarbazide - EPR Electron Paramagnetic Resonance - FeCN potassium ferricyanide - HEPES N-2-hydroxyethylenepiperazine-N-2-ethanesulfonate - P680 reaction center of PSII - PA Photoacoustic - Pheo pheophytin - PSI photosystem I - PSII photosystem II - Q_A primary electron acceptor of PSII     

Effects of exercise training and diabetes on cardiac myosin heavy chain composition

This study determined whether the beneficial effects of exercise training on the diabetic heart previously observed are associated with alterations in ventricular myosin heavy chain (MHC) isoform composition. Diabetes was induced in rats by i.v. streptozotocin. Trained rats were run on a treadmill for 60 min/day, 27 m/min, 10% grade. After 10 wks, ventricular MHC isoenzyme protein composition was analyzed for MHC composition using gel electrophoresis. -MHC and -MHC mRNA were determined by Northern and slot blot hybridization techniques. Both protein and mRNA analyses indicated that sedentary control rats exhibited a predominance of -MHC. Sedentary diabetics exhibited a shift to -MHC. Exercise trained diabetic rats showed a predominance of -MHC. The results indicate that treadmill exercise training of diabetic rat does not prevent the diabetes-induced shift in MHC composition towards the -MHC isoform, thus it is unlikely that the beneficial effects of exercise training on the diabetic heart, previously shown, are due to a normalization of the myosin isoform composition.     

Improvement of four sigma analysis for the investigation of oxygen evolution by Photosystem II

We present here an improvement to the analysis of oxygen evolution with four sigma coefficients (4-S) by computing z, the sum of the S-state probabilities, which was introduced earlier (Delrieu and Rosengard 1987, Biochim Biophys Acta 892: 163–171). We demonstrate that z is equal to the ratio of two consecutive Mean Y (the estimation of the steady state oxygen production based on local properties) found by three sigma analysis. The quantity z is useful for computing double-hits, and for showing the inactivation/activation processes of PS II complexes. Three sigma analysis assumes z=1 exactly; since this is not verified, it is argued that four sigma analysis is closer to the real workings of the water oxidizing complex. Oxygen evolution can then be interpreted in the frame of a modified Kok's model where the sum of the probabilities equals z. We therefore suggest that the closer fitting of four sigma analysis to oxygen production data is not simply due to an extra, unnecessary variable, but to the fact that PS II complexes can be inactivated and reactivated under flashing light. Finally, in order to facilitate the use of four sigma analysis, a computer program is made available upon request.     

The refined 2.15 A X-ray crystal structure of human liver cathepsin B: the structural basis for its specificity.

From the lysosomal cysteine proteinase cathepsin B, isolated from human liver in its two-chain form, monoclinic crystals were obtained which contain two molecules per asymmetric unit. The molecular structure was solved by a combination of Patterson search and heavy atom replacement methods (simultaneously with rat cathepsin B) and refined to a crystallographic R value of 0.164 using X-ray data to 2.15 A resolution. The overall folding pattern of cathepsin B and the arrangement of the active site residues are similar to the related cysteine proteinases papain, actinidin and calotropin DI. 166 alpha-carbon atoms out of 248 defined cathepsin B residues are topologically equivalent (with an r.m.s. deviation of 1.04 A) with alpha-carbon atoms of papain. However, several large insertion loops are accommodated on the molecular surface and modify its properties. The disulphide connectivities recently determined for bovine cathepsin B by chemical means were shown to be correct. Some of the primed subsites are occluded by a novel insertion loop, which seems to favour binding of peptide substrates with two residues carboxy-terminal to the scissile peptide bond; two histidine residues (His110 and His111) in this "occluding loop' provide positively charged anchors for the C-terminal carboxylate group of such polypeptide substrates. These structural features explain the well-known dipeptidyl carboxypeptidase activity of cathepsin B. The other subsites adjacent to the reactive site Cys29 are relatively similar to papain; Glu245 in the S2 subsite favours basic P2-side chains. The above mentioned histidine residues, but also the buried Glu171 might represent the group with a pKa of approximately 5.5 near the active site, which governs endo- and exopeptidase activity. The "occluding loop' does not allow cystatin-like protein inhibitors to bind to cathepsin B as they do to papain, consistent with the reduced affinity of these protein inhibitors for cathepsin B compared with the related plant enzymes.     

Morphological changes and induced sporulation in HmBR transformants ofCochliobolus lunatus

The phenomena following the transformation of the fungusCochliobolus lunatus by plasmid-encoded HmB resistance were investigated. All of the 16 tested transformants had markedly altered morphology. Unlike the untransformed fungus, the transformants produced both conidia and arthrospores, did not excrete slime, lost their purple color, and had an altered progesterone-bioconverting pathway.     

Electron Transport-Dependent Chlorophyll-a Fluorescence Quenching by O(2) in Various Algae and Higher Plants

A comparison of chlorophyll-a fluorescence in brown algae (Macrocystis integrifolia, Fucus vesiculosis), green algae (Scenedesmus obliquus, Ulva sp.) and higher plants (bean, corn) show differences in the relative fluorescence intensities and induction time courses which characterize each type of plant. These differences are not reflected in either the maximum fluorescence emission in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (F_max) or the nonvariable fluorescence (F_o). Constancy of F_o and F_max suggests functional similarities of photosystem II and associated antennae pigments in the various classes of plants. The time course differences are observed only in the absence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea and appear, therefore, to be electron transport dependent. During induction, the peak in fluorescence (F_p) is much lower in all of the algae studied than in the higher plants. Exogenous O₂ strongly quenches F_p in all plants studied and our data indicate that the low F_p in the algae can be partially accounted for by endogenous O₂ quenching.     

Differential activation of the mouse beta-globin promoter by enhancers.

A series of plasmids was constructed to study the effect of two enhancers, the simian virus 40 72-base-pair repeat and the Harvey sarcoma virus 73-base-pair repeat, on the mouse beta maj-globin promoter. These plasmids contain the mouse beta maj-globin promoter linked to the Escherichia coli galK gene, thus allowing galactokinase enzyme activity to be used as a measure of promoter function. In CV-1 (primate) cells, it was found that an enhancer is required for optimal promoter activity and that the simian virus 40 (primate) enhancer increases galactokinase fourfold more than the Harvey sarcoma virus (mouse) enhancer. In L (mouse) cells, however, the Harvey sarcoma virus enhancer is 1.3-fold stronger than the simian virus 40 enhancer. These data support the hypothesis that enhancer activity can be species specific. Furthermore, when both enhancers are present on the same plasmid, their effect is additive on the beta-globin promoter whether the plasmid is in CV-1 cells or L cells.     

Importance de l'étude de la répartition écologique pour la taxonomie des éponges calcaires

Résumé 1. Les travaux deSarà (1953) et deBurton (1963) ont remis en question la valeur systématique des caractères morphologiques dans la classification des Eponges Calcaires, et ont proposé de considérer de nombreuses formes connues comme des variétés individuelles et régionales d'espèces très plastiques et variables. Nous avons essayé de préciser, par l'observation directe en plongée libre, la distribution et les préférences écologiques des Calcaires, de définir ainsi les limites naturelles de leurs populations, et d'étudier ensuite la variabilité des caractères morphologiques à l'intérieur de ces populations.2. Nous exposons ici l'exemple des espèces des genresClathrina etAscandra, étudiées en Adriatique, dans les régions de Marseille et de Roscoff. Leur distribution est en rapport direct et très précis avec l'exposition aux vagues et à la lumière, à tel point que sur une surface réduite plusieurs espèces se succèdent et se remplacent en fonction de l'exposition de l'endroit. Dans la règion de Roscoff cette distribution est plus large ce qui est dû aux effets des marées, mais les zones écologique préférentielles pour les espèces deClathrina ont pu être établies en fonction de l'importance de l'apport des sédiments par les eaux côtières.3. La possibilité de définir des populations dont la distribution est très précisemment délimitée malgré une coexistence dans un espace réduit, et qui ne sont pas réliées par les formes transitoires, permet d'éliminer l'hypothèse de la cospécificité de ces populations. Elle permet également d'établir les variations morphologiques possibles au sein d'une espèce, de retenir les caractères significatifs pour la classification, et finalement de distinguer les espèces qui se ressemblent morphologiquement. Dans le cas d'animaux qui, commeClathrina, possèdent très peu de caractères utilisables en systématique, c'est la seule façon de résoudre les problèmes de leur taxonomie.

---

The importance of research on the ecological distribution for the taxonomy of calcareous sponges

The particular difficulties of the taxonomy of calcareous sponges are due to their great plasticity and their capacity for adaptation which obscure the natural limits of species. An attempt was made to define the natural populations of the Mediterranean species of the generaClathrina andAscandra through a study of their ecological distributions. The populations not connected by the transitory forms were defined by their ecological preferences, and, as they are found in a close cohabitation, were treated as having the specific values. The variability of different morphological features is discussed, the features significant for taxonomy are pointed out and the morphologically close species are defined.