Seasonal variation in stable C and N isotope ratios of the Rhone River inputs to the Mediterranean Sea (2004–2005)

The temporal variation in the stable carbon and nitrogen isotope ratios of particulate organic matter (POM) in the Rhone River was investigated on a monthly basis during a 2-year period (2004–2005). In spite of high month-to-month variation, interannually consistent seasonal trends appeared, with significantly lower δ¹³C (<?28.2‰) in spring than in the other seasons. In contrast, δ¹⁵N did not exhibit significant temporal variation. In spring and summer, high chlorophyll a and b concentrations were associated with low C/N values (<8) and a high percentage of organic carbon (%C) and organic nitrogen (%N), testifying to high development of autochthonous riverine phytoplankton (mainly diatoms and chlorophytes). In fall and winter, higher δ¹³C (>?27.2‰) and C/N (>8) values, and lower %C, %N, and chlorophylls concentrations indicated the predominance of allochthonous terrestrial detritus material in the river POM. The lower δ¹³C values recorded in spring–summer, when the phytoplankton biomass was high, were related to the lower carbon isotopic signatures of freshwater diatoms and chlorophytes compared to those of terrestrial plants. Overall, Rhone River POM was mainly composed of terrestrially derived material (90%), with autochthonous phytoplankton representing only 10% as a mean, in spite of a higher mean contribution of phytoplankton (27%) to river POM in summer.     

Downregulation of CK2 induces apoptosis in cancer cells – A potential approach to cancer therapy

We have previously documented that naked antisense CK2α ODN can potently induce apoptosis in cancer cells in culture and in mouse xenograft human prostate cancer. The effects of the antisense CK2α are related to downregulation of CK2α message and rapid loss of the CK2 from the nuclear compartment. Here we demonstrate that downregulation of CK2 elicited by diverse methods leads to inhibition of cell growth and induction of apoptosis. The various approaches to downregulation of CK2 employed were transfection with kinase-inactive plasmid, use of CK2α siRNA, use of inhibitors of CK2 activity, and use of antisense CK2α ODN packaged in sub-50 nm nanocapsules made from tenascin. In all cases, the downregulation of CK2 is associated with loss in cell survival. We have also described preliminary observations on an approach to targeting CK2 in cancer cells. For this, sub-50 nm tenascin-based nanocapsules bearing the antisense CK2α ODN were employed to test that the antisense is delivered to the cancer cells in vivo. The results provide the first preliminary evidence that such an approach may be feasible for targeting CK2 in cancer cells. Together, our results suggest that CK2 is potentially a highly plausible target for cancer therapy.     

What sources of organic carbon drive food webs in billabongs? A study based on stable isotope analysis

We used the stable isotopes of carbon and nitrogen to examine the food webs of three small flood-plain lakes (billabongs) in south-eastern Australia. With few exceptions, stable carbon isotope analysis could not be used to discriminate among the conspicuous potential sources of fringing, emergent or floating vegetation or benthic detritus. These primary sources showed little spatial or temporal variation in ¹³C values, with means ranging from-28.5 to-26.8 in spring and-29.1 to-25.4 in late summer. Submerged vegetation had similar ¹³C values to the above sources in spring but showed greater spatial variation and were less ¹³C-depleted, considerably so in some species, in late summer. Epiphytes and algae were ¹³C-depleted in spring compared with the other primary sources but became more ¹³C-enriched in late summer. Mean ¹³C values for primary and secondary consumers were not only far more variable (-37.4 to-22.7) but in general were more negative than the potential food sources, particularly in spring. Using the combined information from stable carbon and nitrogen isotope analysis, we could narrow down the list of potential primary sources driving food webs in these billabongs. The freshwater crayfish (Cherax) was one of the few taxa that appeared to obtain its biomass carbon from detrital material. Gastropods and leptocerid caddis larvae on emergent or submerged vegetation obtained a mixture of carbon from epiphytes and macrophytes; in both taxa, epiphytes contributed more to biomass carbon than did the macrophytes. However, other common grazers and collector/gatherers sampled from macrophytes, e.g. baetid mayflies, chironomid larvae and atyid shrimps, were often too ¹³C-depleted even to have derived their biomass carbon solely from epiphytes. Many other primary consumers, including zooplankton, and mussels (Velesunio), and most of the secondary consumers, including water mites (Hydracarina), phantom midge larvae (Chaoborus) and fish, were also ¹³C-depleted. The enormous biomass of littoral and fringing vegetation could contribute to metazoan food webs in these billabongs only if an additional highly ¹³C-depleted source was consumed simultaneously. Methane released from billabong sediments could provide such a ¹³C-depleted carbon source that is re-introduced into metazoan food webs via the consumption of methanotrophic bacteria. Alternatively, food webs in these water bodies are largely driven by an unknown and inconspicuous ¹³C-depleted primary producer, such as planktonic Chlorophyta.     

A Bayesian approach to conservation genetics of Blanding’s turtle (Emys blandingii) in Ontario,Canada

Blanding’s turtle, Emys blandingii, is a globally endangered species with a range centred on the Great Lakes of North America. Several disjunct populations also occur along the East Coast of North America. Previous studies suggest that the Great Lakes portion of the species’ range exhibits panmixia. However, E. blandingii is restricted to relatively small populations in many areas around the Great Lakes. Therefore, panmixia across large geographic distances in this area is unlikely. Here, we apply Bayesian analyses of population structure to samples collected across southern Ontario (N = 97) to test a null hypothesis of panmixia and assess possible management units (MUs), and to estimate rates of gene flow across the study area. Sampled sites in Ontario represent a minimum of four distinct genetic clusters of E. blandingii, which we recommend should be considered as independent MUs. Preliminary evidence suggests that further structure may be present in less robustly sampled areas, which deserve further consideration. Genetic diversity at sampled sites is comparable to that reported for other freshwater turtles. Our comparison between this study and previous work confirms that genetic diversity in E. blandingii is reduced in disjunct eastern populations compared to populations centred on the Great Lakes. Genetic diversity in E. blandingii is not correlated with latitude, and instead may reflect post-glacial dispersal of this species from multiple Pleistocene glacial refugia.     

Critical Function of γH2A in S-Phase

Phosphorylation of histone H2AX by ATM and ATR establishes a chromatin recruitment platform for DNA damage response proteins. Phospho-H2AX (γH2AX) has been most intensively studied in the context of DNA double-strand breaks caused by exogenous clastogens, but recent studies suggest that DNA replication stress also triggers formation of γH2A (ortholog of γH2AX) in Schizosaccharomyces pombe. Here, a focused genetic screen in fission yeast reveals that γH2A is critical when there are defects in Replication Factor C (RFC), which loads proliferating cell nuclear antigen (PCNA) clamp onto duplex DNA. Surprisingly Chk1, Cds1/Chk2 and the Rad9-Hus1-Rad1 checkpoint clamp, which are crucial for surviving many genotoxins, are fully dispensable in RFC-defective cells. Immunoblot analysis confirms that Rad9-Hus1-Rad1 is not required for formation of γH2A by Rad3/ATR in S-phase. Defects in DNA polymerase epsilon, which binds PCNA in the replisome, also create an acute need for γH2A. These requirements for γH2A were traced to its role in docking with Brc1, which is a 6-BRCT-domain protein that is structurally related to budding yeast Rtt107 and mammalian PTIP. Brc1, which localizes at stalled replication forks by binding γH2A, prevents aberrant formation of Replication Protein A (RPA) foci in RFC-impaired cells, suggesting that Brc1-coated chromatin stabilizes replisomes when PCNA or DNA polymerase availability limits DNA synthesis.     

A multivalent approach to the discovery of long-acting β(2)-adrenoceptor agonists for the treatment of asthma and COPD

A multivalent approach was applied to the design of long-acting inhaled β(2)-adrenoceptor agonists. A series of dimeric arylethanolamines based on the short acting β(2)-adrenoceptor agonist albuterol were prepared, varying the nature and length of the linker between the basic nitrogens. None of the C(2)-symmetric dimers demonstrated increased potency, however dimer 5j, derived from 4-phenethylamine, was found to have increased binding potency in vitro relative to the parent monomer. Optimization of this structure led to the identification of 22 (milveterol) which demonstrates high potency in vitro and long duration of action in a guinea pig model of bronchoprotection.     

A reductionist approach to the analysis of learning in brain–computer interfaces

The complexity and scale of brain–computer interface (BCI) studies limit our ability to investigate how humans learn to use BCI systems. It also limits our capacity to develop adaptive algorithms needed to assist users with their control. Adaptive algorithm development is forced offline and typically uses static data sets. But this is a poor substitute for the online, dynamic environment where algorithms are ultimately deployed and interact with an adapting user. This work evaluates a paradigm that simulates the control problem faced by human subjects when controlling a BCI, but which avoids the many complications associated with full-scale BCI studies. Biological learners can be studied in a reductionist way as they solve BCI-like control problems, and machine learning algorithms can be developed and tested in closed loop with the subjects before being translated to full BCIs. The method is to map 19 joint angles of the hand (representing neural signals) to the position of a 2D cursor which must be piloted to displayed targets (a typical BCI task). An investigation is presented on how closely the joint angle method emulates BCI systems; a novel learning algorithm is evaluated, and a performance difference between genders is discussed.     

A novel approach to identification of somatic retroelements’ insertions in human genome

The activity of retroelements is one of the factors leading to genetic variability of the modern humans. Insertions of retroelements may result in alteration of gene expression and functional diversity between cells. In recent years an increasing amount of data indicating an elevated level of retroelements’ mobilisation in some human and animal tissues has been reported. Therefore, the development of a system for the detection of somatic retroposition events is required. Here we describe a novel approach to the whole-genome identification of somatic retroelement insertions in human genome. The developed approach was applied for the comparisons of somatic mosaicism levels in two tissues of the investigated individual. A total of 3410 insertions of retroelements belonging to AluYa5 subfamily were identified.     

A Supplement to the 《Flora of Xizang》 (2)

The present paper is the Supplement 2 to the Flora of Xizang, based upon a collection in 1980 by Mr. W. L. Chen et al. from Mêdog, the south-eastern part of Xizang. In the paper 11 new species are desribed and 10 new-record species are reported. All the type specimens are kept in the Herbarium of the Institure of Botany, Academia Sinica (PE).     

A quantitative hydrogeomorphic approach to the classification of temporary wetlands in the Doñana National Park (SW Spain)

A quantitative hydrogeomorphic approach was applied to an extensive survey of temporary wetlands in the Doñana National Park (SW Spain) in search for quantitative thresholds for wetland classification. Twenty freshwater ponds on the aeolian sand mantle and 46 sites on silty-clay substrate, located in the southern marshland, were surveyed during the heaviest rainy period so far recorded (October 1995–September 1997). On average, temporary ponds showed higher water depth, longer flooding period, lower conductivity (<0.5 mS cm^?1), lower pH (6.7), lower phosphate concentration (0.4 μM) and a more balanced proportion of Ca²⁺/Na⁺ than temporary marshes. During floods, marshland sites exhibited higher water transparency, pH (9.5), alkalinity (3.5 meq l^?1), conductivity (8.2 mS cm^?1), phosphate concentration (0.7 μM), Na⁺ and Ca²⁺ concentrations (97.2 and 3.5 meq l^?1, respectively) than ponds. Study sites were significantly segregated (ANOSIM test: R = 0.88, P < 0.01, n = 92) in relation to water depth and conductivity. A conductivity of 1.6 mS cm^?1 is proposed as a threshold between marshland sites and ponds during floods. Marshland sites were further segregated into two groups (ANOSIM test: R = 0.777, P < 0.01, n = 23) according to the Na⁺/Ca²⁺ ratio (in meq l^?1) at a threshold value of 25. An ordination by PCA showed that five variables grouped 81.4% of the total variance in two axes. The first PCA axis (60.7% of variance) separated temporary wetlands into ponds and marshland sites according to variables related to substrate and hydrology (Na⁺/Ca²⁺ ratio, conductivity, water depth and flooding period). Other variables (e.g., water transparency, alkalinity, pH, submersed macrophyte biomass, phosphate, nitrate and planktonic chlorophyll concentrations) did not produce a significant segregation between marshland and pond sites during floods. Further discrimination within each wetland type was thus not achieved.     

Binding of β4γ5 by adenosine A1 and A2A receptors determined by stable isotope labeling with amino acids in cell culture and mass spectrometry

Characterization of G protein βγ dimer isoform expression in different cellular contexts has been impeded by low levels of protein expression, broad isoform heterogeneity, and antibodies of limited specificity, sensitivity, or availability. As a new approach, we used quantitative mass spectrometry to characterize native βγ dimers associated with adenosine A(1):α(i1) and adenosine A(2A):α(S) receptor fusion proteins expressed in HEK-293 cells. Cells expressing A(1):α(i1) were cultured in media containing [(13)C(6)]Arg and [(13)C(6)]Lys and βγ labeled with heavy isotopes purified. Heavy βγ was combined with either recombinant βγ purified from Sf9 cells, βγ purified from the A(2A):α(S) expressed in HEK-293 cells cultured in standard media, or an enriched βγ fraction from HEK-293 cells. Samples were separated by SDS-PAGE, protein bands containing β and γ were excised, digested with trypsin, and separated by HPLC, and isotope ratios were analyzed by mass spectrometry. Three β isoforms, β(1), β(2), and β(4), and seven γ isoforms, γ(2), γ(4), γ(5), γ(7), γ(10), γ(11), and γ(12), were identified in the analysis. β(1) and γ(5) were most abundant in the enriched βγ fraction, and this βγ profile was generally mirrored in the fusion proteins. However, both A(2A):α(S) and A(1):α(i1) bound more β(4) and γ(5) compared to the enriched βγ fraction; also, more β(4) was associated with A(2A):α(S) than A(1):α(i1). Both fusion proteins also contained less γ(2), γ(10), and γ(12) than the enriched βγ fraction. These results suggest that preferences for particular βγ isoforms may be driven in part by structural motifs common to adenosine receptor family members.     

Did Media Attention of the 2009 A(H1N1) Influenza Epidemic Increase Outpatient Antibiotic Use in France?: A Time-Series Analysis

Background

In France, the 2009 A(H1N1) influenza epidemic occurred between September 2009 and January 2010. Sparking widespread controversy, it was intensely reported in the media. Despite therapeutic inefficacy, antibiotic consumption and viral respiratory infections are positively correlated, particularly in France, where antibiotic overconsumption is well-known. We first determined the period when media coverage was high, and then compared, during this period, observed outpatient antibiotic consumption to estimated outpatient antibiotic consumption “without media attention”.

Materials and Methods

To evaluate media coverage, two online databases were consulted: Factiva and Europresse. To quantify outpatient antibiotic consumption, we used data on reimbursements of outpatient systemic antibiotics from the computerized databases of the two main National Health Insurance agencies. Influenza-like syndromes data came from the French GPs Sentinelles Network. Weekly time-series of antibiotic consumption were modeled by autoregressive moving-average models with exogenous inputs and interventions. Analyses were computed for the entire series and by age group (0–5, 6–15, 16–60, and >60 years).

Results

Media coverage was intense between April 2009 and January 2010. No effect on total outpatient antibiotic consumption was observed during the whole mediatic period. However, during the epidemic in France (September 2009-January 2010), we found an antibiotic underconsumption for the entire series, 0–5 and >60 years. Additionally, at the beginning of the pandemic, when cases were still outside France (June 2009-August 2009), we found an antibiotic overconsumption for patients >16 years.

Conclusion

The early period of A(H1N1) virus circulation compared with seasonal influenza or an overdeclaration of ILS cases might explain the antibiotic underconsumption observed during the period of active A(H1N1) virus transmission in France. At the pandemic onset, when uncertainty was high, the overconsumption observed for individuals >16 years might have been caused by alarmist media reporting. Additional analyses are needed to understand the determinants of antibiotic consumption during this period.     

Airborne pollen of Ambrosia in Burgundy (France) 1996–1997

Pollen counts in Burgundy were monitored by means of four Hirst volumetric traps. Their analysis revealed a sharp rise of Ambrosia from 1996 to 1997. The minimum amount of ragweed pollen likely to provoke allergies (13 grains m–3) was reached or exceeded several days every year, but without leading to clinical symptoms. The circadian variation of Ambrosia pollen showed two peaks: the first one in the morning was suggested to coincide with local pollination, although the second one, in the afternoon, seemed to result from a long range transport which was confirmed by the study of wind roses.     

A chemoenzymatic approach to the synthesis of enantiomerically pure (S)-3-hydroxy-γ-butyrolactone

Optically pure (S)-3-hydroxy-gamma-butyrolactone, an important chiral building block in the pharmaceutical industry, was synthesized from L: -malic acid by combining a selective hydrogenation and a lipase-catalyzed hydrolysis. Lipase from Candida rugosa was found to be the most efficient enzyme for the hydrolysis of (S)-beta-benzoyloxy-gamma-butyrolactone. The use of organic solvent-aqueous two-phase system was employed to extract benzoic acid generated from enzymatic hydrolysis of the substrate. Tert-butyl methyl ether as an organic solvent was effective to extract the reaction product, benzoic acid, and stably maintained the enzyme activity of Lipase OF immobilized on polymeric supports Amberlite XAD-7. The immobilization made the recovery of the product simpler and prevented the formation of the emulsion. The pH adjustment was unnecessary with the immobilized Lipase OF. The scale-up of the enzymatic hydrolysis of S-BBL at 1,850-kg scale was carried out without problems to give 728.5 kg of S-HGB at 80% isolated yield. The scale-up results are similar to those of bench scale reactions. Racemic (R,S)-beta-benzoyloxy-gamma-butyrolactone was prepared from D-, L: -malic acid and was found to be hydrolyzed nonselectively by the enzyme.     

A phylogenetic and allozyme approach to understanding rarity in three “green ash” eucalypts (Myrtaceae)

Phylogenetic relationships among the 12 species of the green ash group of eucalypts were examined using allozyme data, to investigate the causes of rarity in three localized endemics of the group. The relationships suggested by the allozyme data showed both similarities to and differences from those estimated from morphological data byLadiges and coworkers. The phylogenetic relationships suggest that rarity inEucalyptus burgessiana may be related to recent divergence, whileE. paliformis andE. rupicola are relatively old species, more likely to be relicts, and/or restricted to a rare habitat.     

Why are cyclodisilazane rings more stable than cyclodisiloxanes? A qualitative molecular orbital approach to the bonding in cyclodisilazanes and cyclodisiloxanes

The approximate molecular orbitals of cyclic Si₂E₂ (E=nitrogen or oxygen) rings are discussed. It is shown that, due to high silicon 3p_z orbital contribution to the siloxane HOMO, the 3d orbitals can not strenghten the silicon-oxygen bond. In contrast, in the silazane ring considerable Si(3d_π)N(2pπ) bonding may occur. These additional π bonds are responsible for the relative stability of cyclodisilazane rings as compared with the isoelectronic cyclodisiloxane rings.