Kinetics of phyllosemiquinone oxidation in the Photosystem I reaction centre of Acaryochloris marina

Light-induced electron transfer reactions in the chlorophyll a/d-binding Photosystem I reaction centre of Acaryochloris marina were investigated in whole cells by pump-probe optical spectroscopy with a temporal resolution of ~5ns at room temperature. It is shown that phyllosemiquinone, the secondary electron transfer acceptor anion, is oxidised with bi-phasic kinetics characterised by lifetimes of 88±6ns and 345±10ns. These lifetimes, particularly the former, are significantly slower than those reported for chlorophyll a-binding Photosystem I, which typically range in the 5-30ns and 200-300ns intervals. The possible mechanism of electron transfer reactions in the chlorophyll a/d-binding Photosystem I and the slower oxidation kinetics of the secondary acceptors are discussed.     

Probing the role of chloride in Photosystem II from Thermosynechococcus elongatus by exchanging chloride for iodide

The active site for water oxidation in Photosystem II (PSII) goes through five sequential oxidation states (S(0) to S(4)) before O(2) is evolved. It consists of a Mn(4)CaO(5) cluster and Tyr(Z), a redox-active tyrosine residue. Chloride ions have been known for long time to be required for the function of the enzyme. However, X-ray data have shown that they are located about 7? away from the Mn(4)CaO(5) cluster, a distance that seems too large to be compatible with a direct involvement of chloride in the water splitting chemistry. We have investigated the role of this anion by substituting I(-) for Cl(-) in the cyanobacterium Thermosynechococcus elongatus with either Ca(2+) or Sr(2+) biosynthetically assembled into the Mn(4) cluster. The electron transfer steps affected by the exchanges were investigated by time-resolved UV-visible absorption spectroscopy, time-resolved EPR at room temperature and low temperature cw-EPR spectroscopy. In both Ca-PSII and Sr-PSII, the Cl(-)/I(-) exchange considerably slowed down the two S(3)Tyr(Z)(?)→(S(3)Tyr(Z)(?))'→S(0) reactions in which the fast phase, S(3)Tyr(Z)(?)→(S(3)Tyr(Z)(?))', reflects the electrostatically triggered expulsion of one proton from the catalytic center caused by the positive charge near/on Tyr(Z)(?) and the slow phase corresponds to the S(0) and O(2) formations and to a second proton release. The t(1/2) for S(0) formation increased from 1.1ms in Ca/Cl-PSII to ≈6ms in Ca/I-PSII and from 4.8ms in Sr/Cl-PSII to ≈45ms in Sr/I-PSII. In all cases the Tyr(Z)(?) reduction was the limiting step. The kinetic effects are interpreted by a model in which the Ca(2+) binding site and the Cl(-) binding site, although spatially distant, interact. This interaction is likely mediated by the H-bond and/or water molecules network(s) connecting the Cl(-) and Ca(2+) binding sites by which proton release may be channelled.     

Mobile elements in archaeal genomes

The recent availability of several archaeal genome sequences has provided a basis for detailed analyses of the frequency, location and phylogeny of archaeal mobile elements. All the known elements fall into two main types, autonomous insertion sequence (IS) elements and the non-autonomous miniature inverted repeat element (MITE)-like elements. Both classes are considered to be mobilized via transposases that are encoded by the IS elements, although mobility has only been demonstrated experimentally for a few elements. The number, and diversity, of the elements differs greatly between the genomes. At one extreme Sulfolobus solfataricus P2 and Halobacterium NRC-1 are very rich in elements while Methanobacterium thermoautotrophicum contains none. The former also show examples of complex clusters of interwoven elements. An analysis of the genomic distribution in S. solfataricus suggests that the putative oriC and terC regions act as barriers for the mobility of both IS and MITE-like elements. Moreover, the very high level of truncated IS elements in the genomes of S. solfataricus, Sulfolobus tokodaii and Thermoplasma volcanium suggests that there may be a cellular mechanism for selectively inactivating IS elements at a point when they become too numerous and disadvantageous for the cell. Phylogenetically, archaeal IS elements are confined to 11 of the 17 known families of bacterial and eukaryal IS elements where some generate distinct subgroups. Finally, DNA viruses, plasmids and DNA fragments can also be inserted into, and excised from, archaeal genomes by means of an integrase-mediated mechanism that has special archaeal characteristics.     

Stem Growth and Epicormic Branch Formation in Understorey Beech Trees (Fagus sylvatica L.)

A study of the formation of epicormic branches on suppressedbeech trees revealed developmental sequences that had not hithertobeen studied in depth. By combining architectural analysis andanatomy, it was possible to trace the history of the plant andthus fully understand the ontogenesis of the individuals studied.The main sequences observed were: (1) the formation of moribundtrees that were gradually covered in epicormic branches fromthe base of the stem to the crown following gradual forest closure;(2) the formation of this new crown of epicormic branches wascombined with a very sharp reduction in stem cambial activity(partial rings, if any) and modifications to the ligneous sections;and (3) resumed girth increment following the establishmentof epicormic branches. After describing in detail the methodsand individual variation, we highlight the spatial and temporalpatterns of the phenomena observed. Lastly, we put forward functionalhypotheses with a view to setting these results against a backgroundof forest tree ontogenesis in general. Copyright 2001 Annalsof Botany Company Fagus sylvatica L., beech, architecture, epicormic shoot, anatomy, growth, annual wood ring, missing ring     

When Giants Turn Up: Sighting Trends,Environmental Influences and Habitat Use of the Manta Ray Manta alfredi at a Coral Reef

Manta rays Manta alfredi are present all year round at Lady Elliot Island (LEI) in the southern Great Barrier Reef, Australia, with peaks in abundance during autumn and winter. Drivers influencing these fluctuations in abundance of M. alfredi at the site remain uncertain. Based on daily count, behavioural, weather and oceanographic data collected over a three-year period, this study examined the link between the relative number of sightings of manta rays at LEI, the biophysical environment, and the habitat use of individuals around the LEI reef using generalised additive models. The response variable in each of the three generalised additive models was number of sightings (per trip at sea) of cruising, cleaning or foraging M. alfredi. We used a set of eleven temporal, meteorological, biological, oceanographic and lunar predictor variables. Results for cruising, cleaning and foraging M. alfredi explained 27.5%, 32.8% and 36.3% of the deviance observed in the respective models and highlighted five predictors (year, day of year, wind speed, chlorophyll-a concentration and fraction of moon illuminated) as common influences to the three models. There were more manta rays at LEI in autumn and winter, slower wind speeds, higher productivity, and around the new and full moon. The winter peak in sightings of foraging M. alfredi was found to precede peaks in cleaning and cruising activity around the LEI reef, which suggests that enhanced food availability may be a principal driver for this seasonal aggregation. A spatial analysis of behavioural observations highlighted several sites around the LEI reef as ‘multi-purpose’ areas where cleaning and foraging activities commonly occur, while the southern end of the reef is primarily a foraging area. The use of extensive citizen science datasets, such as those collected by dive operators in this study, is encouraged as they can provide valuable insights into a species'' ecology.     

Early Sorafenib-Induced Toxicity Is Associated with Drug Exposure and UGTIA9 Genetic Polymorphism in Patients with Solid Tumors: A Preliminary Study

Background

Identifying predictive biomarkers of drug response is of key importance to improve therapy management and drug selection in cancer therapy. To date, the influence of drug exposure and pharmacogenetic variants on sorafenib-induced toxicity remains poorly documented. The aim of this pharmacokinetic/pharmacodynamic (PK/PD) study was to investigate the relationship between early toxicity and drug exposure or pharmacogenetic variants in unselected adult outpatients treated with single-agent sorafenib for advanced solid tumors.

Methods

Toxicity was recorded in 54 patients on days 15 and 30 after treatment initiation and sorafenib exposure was assessed in 51 patients. The influence of polymorphisms in CYP3A5, UGT1A9, ABCB1 and ABCG2 was examined in relation to sorafenib exposure and toxicity. Clinical characteristics, drug exposure and pharmacogenetic variants were tested univariately for association with toxicities. Candidate variables with p<0.1 were analyzed in a multivariate analysis.

Results

Gender was the sole parameter independently associated with sorafenib exposure (p = 0.0008). Multivariate analysis showed that increased cumulated sorafenib (AUC_cum) was independently associated with any grade ≥3 toxicity (p = 0.037); UGT1A9 polymorphism (rs17868320) with grade ≥2 diarrhea (p = 0.015) and female gender with grade ≥2 hand-foot skin reaction (p = 0.018). Using ROC curve, the threshold AUC_cum value of 3,161 mg/L.h was associated with the highest risk to develop any grade ≥3 toxicity (p = 0.018).

Conclusion

In this preliminary study, increased cumulated drug exposure and UGT1A9 polymorphism (rs17868320) identified patients at high risk for early sorafenib-induced severe toxicity. Further PK/PD studies on larger population are warranted to confirm these preliminary results.     

Bloom's syndrome and PICH helicases cooperate with topoisomerase IIα in centromere disjunction before anaphase

Centromeres are specialized chromosome domains that control chromosome segregation during mitosis, but little is known about the mechanisms underlying the maintenance of their integrity. Centromeric ultrafine anaphase bridges are physiological DNA structures thought to contain unresolved DNA catenations between the centromeres separating during anaphase. BLM and PICH helicases colocalize at these ultrafine anaphase bridges and promote their resolution. As PICH is detectable at centromeres from prometaphase onwards, we hypothesized that BLM might also be located at centromeres and that the two proteins might cooperate to resolve DNA catenations before the onset of anaphase. Using immunofluorescence analyses, we demonstrated the recruitment of BLM to centromeres from G2 phase to mitosis. With a combination of fluorescence in situ hybridization, electron microscopy, RNA interference, chromosome spreads and chromatin immunoprecipitation, we showed that both BLM-deficient and PICH-deficient prometaphase cells displayed changes in centromere structure. These cells also had a higher frequency of centromeric non disjunction in the absence of cohesin, suggesting the persistence of catenations. Both proteins were required for the correct recruitment to the centromere of active topoisomerase IIα, an enzyme specialized in the catenation/decatenation process. These observations reveal the existence of a functional relationship between BLM, PICH and topoisomerase IIα in the centromere decatenation process. They indicate that the higher frequency of centromeric ultrafine anaphase bridges in BLM-deficient cells and in cells treated with topoisomerase IIα inhibitors is probably due not only to unresolved physiological ultrafine anaphase bridges, but also to newly formed ultrafine anaphase bridges. We suggest that BLM and PICH cooperate in rendering centromeric catenates accessible to topoisomerase IIα, thereby facilitating correct centromere disjunction and preventing the formation of supernumerary centromeric ultrafine anaphase bridges.     

Resident CD11b+Ly6C? Lung Dendritic Cells Are Responsible for Allergic Airway Sensitization to House Dust Mite in Mice

Conventional dendritic cells (DCs) are considered to be the prime initiators of airway allergy. Yet, it remains unclear whether specific DC subsets are preferentially involved in allergic airway sensitization. Here, we systematically assessed the respective pro-allergic potential of individually sorted lung DC subsets isolated from house dust mite antigen (HDM)-treated donor mice, following transfer to naïve recipients. Transfer of lung CD11c⁺CD11b⁺ DCs, but not CD11c⁺CD11b⁻CD103⁺ DCs, was sufficient to prime airway allergy. The CD11c⁺CD11b⁺ DC subpopulation was composed of CD11c⁺CD11b⁺Ly6C⁺ inflammatory monocyte-derived cells, whose numbers increase in the lungs following HDM exposure, and of CD11c⁺CD11b⁺Ly6C⁻ DCs, which remain stable. Counterintuitively, only CD11c⁺CD11b⁺Ly6C⁻ DCs, and not CD11c⁺CD11b⁺Ly6C⁺ DCs, were able to convey antigen to the lymph nodes and induce adaptive T cell responses and subsequent airway allergy. Our results thus support that lung resident non-inflammatory CD11c⁺CD11b⁺Ly6C⁻ DCs are the essential inducers of allergic airway sensitization to the common aeroallergen HDM in mice.