Foregut Development and Metamorphosis in a Pyramidellid Gastropod: Modularity and Constraint within a Complex Life Cycle.

Pyramidellids are tiny ectoparasitic gastropods with highly derived feeding structures for piercing and sucking. We attempted to resolve homology controversies about unique pyramidellid feeding structures by examining foregut development through larval and metamorphic stages, using sections for light and electron microscopy. We anticipated that, like many marine invertebrate larvae, post-metamorphic structures would differentiate extensively in late larvae to speed metamorphic transition. Previous studies of gastropods suggested that development of juvenile feeding structures in larvae was facilitated by foregut subdivision into dorsal and ventral developmental modules, and spatial uncoupling of these modules may have facilitated adaptive radiation in neogastropods. Observations of Odostomia tenuisculpta suggested that the stylet may be derived from cuticle-secreting buccal epithelium surrounding the proximal end of the salivary duct, whereas the stylet sheath could be either a derived jaw or a radular tooth. The anterior half of the remarkable buccal pump of these euthyneuran gastropods develops from the larval esophagus, which is unorthodox compared to caenogastropods, where extensive post-metamorphic specialization of a dorsal module component has not been previously described. The introvert tube develops from pouches of the distal larval esophagus and may actually be an eversible oral tube rather than an acrembolic proboscis. Minimal differentiation of presumptive juvenile foregut structures occurred during the larval stage of O. tenuisculpta, when compared to other gastropods. The stylet, stylet sheath, and buccal pump may be incompatible with functioning of the larval esophagus; thus, an explosive period of morphogenesis is necessary at metamorphosis. Although dorsal and ventral modules were recognizable during the development of O. tenuisculpta, we failed to find evidence that this modularity facilitated the extreme evolutionary remodeling of post-metamorphic feeding structures.

The Lewis superacid Al[N(C6F5)2]3 and its higher homolog Ga[N(C6F5)2]3 - structural features, theoretical investigation and reactions of a metal amide with higher fluoride ion affinity than SbF5.

Herein we present the synthesis of the two Lewis acids Al[N(C6F5)2]3 (ALTA) and Ga[N(C6F5)2]3 (GATA) via salt elimination reactions. The metal complexes were characterized by NMR-spectroscopic methods and X-ray diffraction analysis revealing the stabilization of the highly Lewis acidic metal centers by secondary metal-fluorine contacts. The Lewis acidic properties of Al[N(C6F5)2]3 and Ga[N(C6F5)2]3 are demonstrated by reactions with Lewis bases resulting in the formation of metallates accompanied by crucial structural changes. The two metallates [Cs(Tol)3]+[FAl(N(C6F5)2)3]- and [AsPh4]+[ClGa(N(C6F5)2)3]- contain interesting weakly coordinating anions. The reaction of Al[N(C6F5)2]3 with trityl fluoride yielded [CPh3]+[FAl(N(C6F5)2)3]- which could find application in the activation of metallocene polymerization catalysts. The qualitative Lewis acidity of Al[N(C6F5)2]3 and Ga[N(C6F5)2]3 was investigated by means of competition experiments for chloride ions in solution. DFT calculations yielded fluoride ion affinities in the gas phase (FIA) of 555 kJ mol-1 for Al[N(C6F5)2]3 and 472 kJ mol-1 for Ga[N(C6F5)2]3. Thus, Al[N(C6F5)2]3 can be considered a Lewis superacid with a fluoride affinity higher than SbF5 (493 kJ mol-1) whereas the FIA of the corresponding gallium complex is slightly below the threshold to Lewis superacidity.

Functional characterisation and cell specificity of BvSUT1, the transporter that loads sucrose into the phloem of sugar beet (Beta vulgaris L.) source leaves.

Sugar beet (Beta vulgaris L.) is one of the most important sugar-producing plants worldwide and provides about one third of the sugar consumed by humans. Here we report on molecular characterisation of the BvSUT1 gene and on the functional characterisation of the encoded transporter. In contrast to the recently identified tonoplast-localised sucrose transporter BvTST2.1 from sugar beet taproots, which evolved within the monosaccharide transporter (MST) superfamily, BvSUT1 represents a classical sucrose transporter and is a typical member of the disaccharide transporter (DST) superfamily. Transgenic Arabidopsis plants expressing the ß-GLUCURONIDASE (GUS) reporter gene under control of the BvSUT1-promoter showed GUS histochemical staining of their phloem; an anti-BvSUT1-antiserum identified the BvSUT1 transporter specifically in phloem companion cells. After expression of BvSUT1 cDNA in bakers' yeasts (Saccharomyces cerevisiae) uptake characteristics of the BvSUT1 protein were studied. Moreover, the sugar beet transporter was characterised as a proton-coupled sucrose symporter in Xenopus laevis oocytes. Our findings indicate that BvSUT1 is the sucrose transporter that is responsible for loading of sucrose into the phloem of sugar beet source leaves delivering sucrose to the storage tissue in sugar beet taproot sinks.

Phylogenetic turnover along local environmental gradients in tropical forest communities.

While the importance of local-scale habitat niches in shaping tree species turnover along environmental gradients in tropical forests is well appreciated, relatively little is known about the influence of phylogenetic signal in species' habitat niches in shaping local community structure. We used detailed maps of the soil resource and topographic variation within eight 24-50 ha tropical forest plots combined with species phylogenies created from the APG III phylogeny to examine how phylogenetic beta diversity (indicating the degree of phylogenetic similarity of two communities) was related to environmental gradients within tropical tree communities. Using distance-based redundancy analysis we found that phylogenetic beta diversity, expressed as either nearest neighbor distance or mean pairwise distance, was significantly related to both soil and topographic variation in all study sites. In general, more phylogenetic beta diversity within a forest plot was explained by environmental variables this was expressed as nearest neighbor distance versus mean pairwise distance (3.0-10.3 % and 0.4-8.8 % of variation explained among plots, respectively), and more variation was explained by soil resource variables than topographic variables using either phylogenetic beta diversity metric. We also found that patterns of phylogenetic beta diversity expressed as nearest neighbor distance were consistent with previously observed patterns of niche similarity among congeneric species pairs in these plots. These results indicate the importance of phylogenetic signal in local habitat niches in shaping the phylogenetic structure of tropical tree communities, especially at the level of close phylogenetic neighbors, where similarity in habitat niches is most strongly preserved.

Smallest molecular chalcogenidometalate anions of the heaviest metals: syntheses, structures, and their interconversion.

The syntheses of the first molecular meta-selenidomercurate(ii), ortho-telluridothallate(iii) and a hydrate of an ortho-selenidoplubate(iv) are presented alongside an improved and facile synthesis of the selenidobismuthate(iii) with almost quantitative yields. By means of quantum chemical calculations, the energetics of the interconversions of small metalate anions is discussed and the existence of the heaviest homologues of [NO2](-), [NO3](-), [PO4](2-) and [CO3](2-) are predicted.

Habitat filtering across tree life stages in tropical forest communities.

Tropical tree communities are shaped by local-scale habitat heterogeneity in the form of topographic and edaphic variation, but the life-history stage at which habitat associations develop remains poorly understood. This is due, in part, to the fact that previous studies have not accounted for the widely disparate sample sizes (number of stems) that result when trees are divided into size classes. We demonstrate that the observed habitat structuring of a community is directly related to the number of individuals in the community. We then compare the relative importance of habitat heterogeneity to tree community structure for saplings, juveniles and adult trees within seven large (24-50 ha) tropical forest dynamics plots while controlling for sample size. Changes in habitat structuring through tree life stages were small and inconsistent among life stages and study sites. Where found, these differences were an order of magnitude smaller than the findings of previous studies that did not control for sample size. Moreover, community structure and composition were very similar among tree sub-communities of different life stages. We conclude that the structure of these tropical tree communities is established by the time trees are large enough to be included in the census (1 cm diameter at breast height), which indicates that habitat filtering occurs during earlier life stages.

Phenotypic and genotypic bacterial antimicrobial resistance in liquid pig manure is variously associated with contents of tetracyclines and sulfonamides.

AIMS: Antibiotic residues as well as antibiotic-resistant bacteria in environmental samples might pose a risk to human health. This study aimed to investigate the association between antibiotic residues and bacterial antimicrobial resistance in liquid pig manure used as fertilizer. METHODS AND RESULTS: Concentrations of tetracyclines (TETs) and sulfonamides (SULs) were determined by liquid chromatography-mass spectrometry in 305 pig manure samples; antibiotic contents were correlated to the phenotypic resistance of Escherichia coli (n = 613) and enterococci (n = 564) towards up to 24 antibiotics. In 121 samples, the concentration of the TET resistance genes tet(M), tet(O) and tet(B) was quantified by real-time-PCR. TETs were found in 54% of the samples. The median sum concentration of all investigated TETs in the positive samples was 0.73 mg kg(-1). SULs were found with a similar frequency (51%) and a median sum concentration of 0.15 mg kg(-1) in the positive samples. Associated with the detection of TETs and/or SULs, resistance rates were significantly elevated for several substances - some of them not used in farm animals, e.g. chloramphenicol and synercid. In addition, multiresistant isolates were found more often in samples containing antibiotics. Analysis of the resistance genes tet(M) and tet(O) already showed a significant increase in their concentrations - but not in tet(B) - in the lowest range of total TET concentration. Mean tet(M) concentrations increased by the factor of 4.5 in the TET concentration range of 0.1-1 mg kg(-1), compared to negative manure samples. CONCLUSIONS: Antibiotic contamination of manure seems to be associated with a variety of changes in bacterial resistance, calling for a prudent use of antibiotics in farm animals. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides an interdisciplinary approach to assess antimicrobial resistance by combining the microbiological analysis of bacterial resistance with high quality chemical analysis of antibiotic residues in a representative number of environmental samples.

Prognosis of women with pT4b breast cancer: the significance of this category in the TNM system.

AIMS: The T4b/pT4b category of the TNM System for breast cancer is discussed controversially. For a more detailed analysis, we explored the prognosis of patients with breast cancer strictly fulfilling the criteria for T4b/pT4b tumors according to the TNM System. METHODS: Retrospectively analysed data from patients with pT4b breast tumors diagnosed between January 1994 and December 2004 were collected. Reclassification was undertaken according to the TNM System criteria establishing a study group including only "correctly" classified T4b/pT4b tumors. A control group with pT1-3 carcinomas was used for analysing the prognostic value of criteria for T4b/pT4b tumors. RESULTS: Eighty-six patients with pT4b carcinomas were found. After reclassification, 65 remained as pT4b fulfilling the strict criteria. The study group showed a 60% three-year Disease Specific Survival (DSS). Age (p<0.01) and regional lymph node status (p<0.01) were significantly related to prognosis. Compared to the control group, the DSS in the study group of patients with a tumor size >2 cm to 5 cm was significantly worse (three-year survival: 82% vs. 51%, p<0.01). For tumors >5 cm, the DSS was not significantly different between both groups (three-year survival: 68% vs. 72%, p=0.7). CONCLUSIONS: The criteria for T4b/pT4b breast cancer are associated with a poorer prognosis in patients with a tumor size >2 cm to 5 cm. For tumors >5 cm, prognosis is independent of T4b/pT4b criteria. These findings do not justify the demanded deletion of the T4b/pT4b category. The missing uniformity in applying the correct criteria of T4b/pT4b tumors queries the practicability of this category.

Are functional traits good predictors of demographic rates? Evidence from five neotropical forests.

A central goal of comparative plant ecology is to understand how functional traits vary among species and to what extent this variation has adaptive value. Here we evaluate relationships between four functional traits (seed volume, specific leaf area, wood density, and adult stature) and two demographic attributes (diameter growth and tree mortality) for large trees of 240 tree species from five Neotropical forests. We evaluate how these key functional traits are related to survival and growth and whether similar relationships between traits and demography hold across different tropical forests. There was a tendency for a trade-off between growth and survival across rain forest tree species. Wood density, seed volume, and adult stature were significant predictors of growth and/or mortality. Both growth and mortality rates declined with an increase in wood density. This is consistent with greater construction costs and greater resistance to stem damage for denser wood. Growth and mortality rates also declined as seed volume increased. This is consistent with an adaptive syndrome in which species tolerant of low resource availability (in this case shade-tolerant species) have large seeds to establish successfully and low inherent growth and mortality rates. Growth increased and mortality decreased with an increase in adult stature, because taller species have a greater access to light and longer life spans. Specific leaf area was, surprisingly, only modestly informative for the performance of large trees and had ambiguous relationships with growth and survival. Single traits accounted for 9-55% of the interspecific variation in growth and mortality rates at individual sites. Significant correlations with demographic rates tended to be similar across forests and for phylogenetically independent contrasts as well as for cross-species analyses that treated each species as an independent observation. In combination, the morphological traits explained 41% of the variation in growth rate and 54% of the variation in mortality rate, with wood density being the best predictor of growth and mortality. Relationships between functional traits and demographic rates were statistically similar across a wide range of Neotropical forests. The consistency of these results strongly suggests that tropical rain forest species face similar trade-offs in different sites and converge on similar sets of solutions.

Tissue inhibitor of metalloproteinases-3 facilitates Fas-mediated neuronal cell death following mild ischemia.

Tissue inhibitor of metalloproteinase-3 (TIMP-3) is a natural inhibitor of metalloproteinases involved in matrix degradation and ectodomain shedding of many cell-surface proteins, including death receptors and/or their ligands. In the present study, we examined the role of TIMP-3 in Fas-mediated neuronal cell death following cerebral ischemia, using both gene deletion and pharmacological approaches. In culture, exposure of primary cortical neurons to 2 h of oxygen-glucose deprivation (OGD) resulted in delayed neuronal cell death that was dependent on activation of the death receptor, Fas. Cortical cultures derived from timp-3(-/-) mice displayed partial resistance against OGD-induced neuronal cell death and also displayed increased shedding of Fas ligand (FasL) into the culture media, compared to wild-type control cultures. Both the increased neuroprotection and increased FasL shedding in timp-3(-/-) cultures were reversed by addition of exogenous metalloproteinase inhibitors, recombinant TIMP-3 or GM6001. In vivo, timp-3(-/-) mice showed marked resistance to a brief (30 min) middle cerebral artery occlusion (MCAO), but were not protected against more severe lesions induced by 90 min of MCAO. These studies demonstrate that TIMP-3 facilitates Fas-mediated neuronal cell death following OGD and plays a pro-apoptotic role in mild cerebral ischemia.

The common and distinct target genes of the p53 family transcription factors.

p53 is the most commonly mutated gene in human cancer. After activation by cellular stresses such as DNA damage or oncogene activation, p53, a sequence-specific DNA-binding protein, induces the expression of target genes which mediate tumor suppression. Two recently identified p53 homologues, p63 and p73, appear to function similarly to p53, that is, they both activate target gene expression and suppress cell growth when overexpressed; however, the p63 and p73 genes are rarely mutated in human cancer and do not adhere to Knudson's classical model of a tumor suppressor gene. Recently, exciting observations suggest nonoverlapping functions for the family members. Herein, we outline the recent literatures identifying and characterizing both the common and distinct target genes of the p53 family transcription factors in relation to their signaling pathways.

The [2+2]-photocycloaddition of aromatic aldehydes and ketones to 3,4-dihydro-2-pyridones: regioselectivity, diastereoselectivity, and reductive ring opening of the product oxetanes.

3,4-Dihydro-2-pyridones [3,4-Dihydropyridin-2(1H)-ones] 6 were evaluated with respect to their use as alkene components in stereoselective Paternò-Büchi reactions. The parent compound 6a was shown to be a versatile synthetic building block that reacted with various photoexcited aromatic carbonyl compounds (benzaldehyde, benzophenone, acetophenone, methyl phenylglyoxylate, 3-pivaloyloxybenzaldehyde) with high regio- and diastereo-selectivity (51-63% yield). The products can be subjected to hydrogenolysis, opening a new and efficient route for the synthesis of 2-arylmethyl-3-piperidinols. As examples, the oxetanes 7a and 8a were hydrogenolytically cleaved and yielded the products 12 (88%) and 13 (93%). The ability of compound 6a to bind to a chiral lactam host through two hydrogen bonds was used favorably to differentiate the enantiotopic faces of its double bond. In the photocycloaddition to the chiral aldehyde 15, which was conducted at -10 degrees C in toluene, a high facial diastereoselectivity (>90% de. 56% yield) was recorded. The stereoselectivity results from a 1:1 association of dihydropyridone 6a to the aldehyde. The 4-substituted dihydropyridones 6b-6d (R = methyl, isopropyl, phenyl) were found to be less suited for potential use in photochemistry. The yields and facial diastereoselectivities recorded in their photocycloaddition to benzophenone remained low.

Synthesis and evaluation of diazine containing bioisosteres of (-)-ferruginine as ligands for nicotinic acetylcholine receptors.

In this structure-affinity relationship (SAFIR) study, the bioisosteric potential of diazines in the field of ferruginine-type nAChR ligands was investigated. Novel enantiopure analogues of (-)-Ferruginine (3) such as 6-8 were synthesized utilizing enantiomerically pure N-protected (+)-2-tropanone 9 from the 'chiral pool' as versatile chiral building block and a palladium-catalyzed Stille cross-coupling of the tributylstannyl diazines 12, 14 and 16 with the vinyl triflate 11 of (+)-2-tropanone 9. The structures of the novel diazine analogues 6-8 of (-)-ferruginine (3) were assigned on the basis of spectral data, that of ligand 7 being additionally verified by X-ray crystallography. The bioisosteric replacement of the acetyl moiety as structural part of the lead compound 3 with the pyridazine, pyrimidine and pyrazine nucleus resulted in ligands with high to moderate affinity for the central alpha4beta2 and remarkably low affinity for the alpha7* nAChR subtypes. Among the compounds synthesized and tested, 7 was the most active one with K(i)=3.7 nM (alpha4beta2). Compared with the lead 3, this value represents a 30-fold improvement in the affinity for the alpha4beta2 subtype combined with a substantially improved selectivity ratio between the alpha4beta2 and alpha7* subtypes.

Phosphabenzenes as monodentate pi-acceptor ligands for rhodium-catalyzed hydroformylation.

A new class of phosphinine/rhodium catalysts for the hydroformylation of terminal and internal alkenes is presented in this study. A series of phosphabenzenes 1-14 has been prepared by condensation of phosphane or tris(trimethylsilyl)phosphane with the corresponding pyrylium salt. Trans-[(phosphabenzene)2RhCl(CO)] complexes 21-25 have been prepared and studied spectroscopically and by X-ray crystal-structure analysis. The hydroformylation of oct-1-ene has been used to identify optimal catalyst preformation and reaction conditions. Hydroformylation studies with 15 monophosphabenzenes have been performed. The catalytic performance is dominated by steric influences, with the phosphabenzene 8/rhodium system being the most active catalyst. Turnover frequencies of up to 45370 h(-1) for the hydroformylation of oct-1-ene have been determined. In further studies, hydroformylation activity toward more highly substituted alkenes was investigated and compared with the standard industrial triphenylphosphane/rhodium catalyst. The reactivity differences between the phosphabenzene and the triphenylphosphane catalyst increase on going to the more highly substituted alkenes. Even tetrasubstituted alkenes reacted with the phosphabenzene catalyst, whereas the triphenylphosphane system failed to give any product. In situ pressure NMR experiments have been performed to identify the resting state of the catalyst. A monophosphabenzene complex [(phosphinine 8)Ir(CO)3H] could be detected as the predominant catalyst resting state.

Intramolecular iron(II)-catalyzed nitrogen transfer reactions of unsaturated alkoxycarbonyl azides: a facile and stereoselective route to 4,5-disubstituted oxazolidinones.

Intramolecular FeII-catalyzed reactions of various unsaturated alkoxycarbonyl azides are described. The reactions occur in the presence of stoichiometric amounts of trimethyl silyl chloride employing ethanol as the solvent. The corresponding 2-alkenyloxycarbonyl azides 5, 9, 18, 20, 22, and 24 gave the products 7/8, 10/11, 19, 21, 23, and 25 of an olefin chloroamination in moderate to good yields (47-72%). The facial diastereoselectivity of the ring closing C-N-bond forming step is good both in cyclic (20, 24) and in acyclic substrates (5, 18, 22) (>90% ds). The subsequent chlorine atom transfer occurs selectively in cyclic systems (20, 24) and in systems (9b, 18) which exhibit a conformational bias in the postulated radical intermediate 14. The lifetime tau of this elusive intermediate was estimated from the loss of stereochemical information in conformationally unrestricted systems (9a, 22) and from the data obtained with a radical clock (31-->32). 2-Alkynyloxycarbonyl azides 34 and 36 also yield chloroamination products which are obtained exclusively as the (Z)-isomers 35 and 37 (81-99% yield). The products of the tert-butyl-substituted substrates 38 undergo an immediate rearrangement/solvolysis reaction in the reaction mixture and gave the 5-alkoxyoxazolidinones 39 (93-99% yield).

Effect of continuous antistaphylococcal therapy on the rate of P. aeruginosa acquisition in patients with cystic fibrosis.

SUMMARY. Continuous therapy with antistaphylococcal antibiotics is advocated by some cystic fibrosis (CF) centers, but it is unclear whether this strategy favors early colonization with P. aeruginosa. We used the data base for the German Centers of the European Registry for Cystic Fibrosis (ERCF) to assess the effect of continuous antistaphyloccocal therapy on the rate of P. aeruginosa acquisition in CF patients. Patients included in this analysis had to be < 18 years of age, P. aeruginosa-negative prior to entry in the ERCF, and to have had at least 2 additional P. aeruginosa-negative respiratory cultures while followed in the ERCF. Of the 639 patients fulfilling these criteria, 48.2% received continuous antistaphyloccocal therapy, 40.4% intermittent antibiotic therapy, and 11.4% no antibiotic therapy. There were no differences between the groups in body mass index, as well as forced vital capacity (FVC) and forced expired volume in 1 sec (FEV(1)) at baseline. The rate at which patients acquired positive respiratory cultures for Staph. aureus was significantly lower in the group receiving continuous antistaphyloccocal antibiotic therapy than in those not receiving such therapy. Patients receiving continuous antistaphyloccocal antibiotic therapy had a significantly higher rate of P. aeruginosa acquisition compared to patients receiving only intermittent or no antibiotic therapy. This difference was especially apparent for children younger than age 6 years. We conclude that continuous therapy with antistapyloccocal antibiotics directed against Staph. aureus increases the risk of colonization with P. aeruginosa. How this affects the clinical outcome of these patients remains to be determined.

Enantioselective photochemical reactions of 2-pyridones in solution.

[reaction: see text] Two key photochemical reactions of prochiral 2-pyridones were studied in the presence of a chiral host. The [4 + 4]-photocycloaddition with cyclopentadiene (CpH) proceeded smoothly and with high enantioselectivity (84-87% ee). The absolute configuration of the endo-diastereoisomer was established by X-ray crystallography. The electrocyclic [4pi]-ring closure to 3-oxo-2-azabicyclo[2.2.0]-5-hexenes occurred with lower enantioselectivity (20-23% ee at -20 degrees C). The velocity of the latter reaction slowed significantly with decreasing temperature.