Acetogenic anthraquinones: biosynthetic convergence and chemical evidence of enzymatic cooperation in nature

Phenylanthraquinones belong to the quite rare class of fully unsymmetric biaryls, consisting of two different molecular portions, an anthraquinone part, chrysophanol, and a phenyl part, 2,4-dihydroxy-6-methoxyacetophenone, linked together by phenol-oxidative coupling. The biosynthesis of these two moieties, from eight and four acetate units, respectively, bears some unique features: Chrysophanol is the first example of an acetogenic natural product that is, in an organism-specific manner, formed via more than one folding mode: In eukaryotes, like, e.g., in fungi, in higher plants, and in insects, it is formed via folding mode F, while in prokaryotes it originates through mode S. It has, more recently, even been found to be synthesized by a third pathway, which we have named mode S′. It is thus the first example of biosynthetic convergence in polyketide biosynthesis. The monocyclic “southern” portion of the molecule, which is much simpler (arising from only four acetate units and without decarboxylation), unexpectedly does not show the anticipated randomization of the C₂-labeling in the aromatic ring, but has clearly localized C₂ units, excluding any symmetric intermediate like, e.g., 2,4,6-trihydroxyacetophenone. This is confirmed by competitive feeding experiments with specifically ¹³C₂-labeled acetophenones, showing the O-methylation to be the decisive symmetry-preventing step, which hints at a close collaboration of the participating enzymes. The results make knipholone an instructive example of structure, function, and evolution of polyketide synthases and O-methyltransferases, and their cooperation.

Purification and Characterization of Four New Cysteine Endopeptidases From Fruits of <Emphasis Type="Italic">Bromelia pinguin</Emphasis> L. Grown in Cuba

Bromelia pinguin L. is a plant broadly distributed in Central America and Caribbean islands. The fruits have been used in traditional medicine as anthelmintic, probably owed to the presence of a mixture of cysteine endopeptidases, initially termed pinguinain. This work deals with the purification and characterization of the four main components of that mixture, two of them showing acid pI and the other two alkaline pI. Molecular masses (SDS-PAGE and MALDI-TOF), N-terminal sequence and the reactivity and kinetic parameters versus synthetic substrates (p-nitrophenyl-N-α-CBZ-amino acid esters, PFLNA, Z-Arg-Arg-p-NA, and Z-Phe-Arg-p-NA) of the studied peptidases are given, as well as the N-terminal sequences of the enzymes and the homology degree with other plant endopeptidases.     

The Upper Cretaceous belemnite <Emphasis Type="Italic">Praeactinocamax plenus</Emphasis> (Blainville, 1827) from Lower Saxony (Upper Cenomanian,northwest Germany) and its distribution pattern in Europe

Occurrences of the Upper Cenomanian (Upper Cretaceous) belemnite Praeactinocamax plenus from the plenus Bed of northwest Germany (Söhlde-Loges working quarry near Salzgitter, Lower Saxony) are documented and described for the first time on the basis of two in situ finds. The find horizon and its surrounding beds are re-evaluated in a sequence stratigraphical context. In contrast to the interpretations of other authors, the plenus Bed is seen as a pelagization event in a parasequence of transgressively stacked beds, delimited by two significant erosion surfaces below and above. The exclusive occurrence of P. plenus in the top part of the plenus Bed and its absence from the post-plenus Bed succession, in the equivalent of which (higher part of the Plenus Marls Member) it is very common in southern England (Anglo-Paris Basin), is explained by ecological factors in stratigraphically complete sections (intra-shelf depressions) and by gaps in the stratigraphic records in swell settings. The distribution pattern of P. plenus suggests a preference for nearshore settings and a demersal mode of life.     

<Emphasis Type="Italic">Noriphyllia</Emphasis>, a new Tethyan Late Triassic coral genus (Scleractinia)

Noriphyllia gen. n. is a distinctive coral representing the Coryphylliidae, a group of Late Triassic scleractinian corals. Coral faunas of this age are poorly known. The new coral is distinguished from related corals belonging to the reimaniphylliids by key features of septal microstructure as discerned in thin sections. This microstructure consists of a straight/wavy midseptal zone and lateral septal stereome organized into thin fascicles of fibres, producing a fine and sharp micromorphology of the septal sides. The solitary genus Noriphyllia gen. n. contains two Early Norian species: N. anatoliensis sp. n. chosen as the type species and N. dachsteinae sp. n., and a Carnian species referred to as N. monotutoensis sp. n. The new genus is widely distributed in the Late Triassic, Early Norian reef facies of the Tethys region (Northern Calcareous Alps, Austria; Taurus Mountains, Turkey) and it also occurs in the Carnian of Timor. Noriphyllia gen. n. is unique and details of its microstructural features add new understanding to the composition of both Late Carnian and Early Norian corals.     

River hydrological seasonality influences life history strategies of tropical riverine fishes

Under a particular set of selective forces, specific combinations of traits (strategies) will be favored in a given population, within the particular constraints of the considered species. For fishes, three demographic strategies have been suggested to result from adaptive responses to environmental predictability (i.e., seasonality): periodic, opportunistic and equilibrium [Winemiller KO, Rose KA (1992) Patterns of life-history diversification in North American fishes: implications for population regulation. Can J Fish Aquat Sci 49:2196–2218]. These strategies optimize fitness within predictable, unpredictable and stable systems, respectively. We tested these predictions of life history trait distribution along a gradient of hydrologic seasonality in West African tropical rivers at the drainage basin scale. We used logistic regression of species presence–absence data to test whether dominant life history traits of species caused community compositional change in response to a gradient of seasonality in hydrologic regime across basins. After accounting for taxonomic relatedness, species body size and statistical redundancy inherent to related traits, we found a higher proportion of species producing a great number of small oocytes, reproducing within a short period of time and presenting a low degree of parental care (the periodic strategy) in highly seasonal drainage basins (e.g., rivers with a short and predictable favorable season). Conversely, in more stable drainage basins (e.g., rivers with a wet season of several months), we observed a greater proportion of species producing small numbers of large oocytes, reproducing within a long period of time and providing parental care to their offspring (the equilibrium strategy). Our results suggest that distributions of tropical freshwater fishes at the drainage basin scale can be partly explained by the match between life history strategies and seasonality gradients in hydrological conditions. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Usefulness of Multiple-Locus VNTR Fingerprinting in detection of clonality of community- and hospital-acquired <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> isolates

Staphylococcus aureus has become a major source of hospital infections and the risk of colonisation and infection by community-acquired methicillin-resistant S. aureus (CA-MRSA) is increasingly higher. Because of the importance of S. aureus to public health, many molecular typing methods have been developed to determine its transmission routes and source of infection during epidemiological investigations. In this study we evaluated the usefulness of multiplex PCR based Multi-Locus VNTR Fingerprinting (MLVF) as the first step method for rapid differentiation of Croatian and Polish S. aureus isolates in hospital and community settings. This is a first report of the usefulness of MLVF in typing of hospital-acquired methicillin-sensitive S. aureus (HA-MSSA) and four CA-MRSA isolates. A total of 47 isolates of S. aureus recovered in Croatia in 2004 and in Poland in 2006 and 2007 were tested. The MLVF results were compared to those produced by other typing methods, such as Pulsed-Field Gel Electrophoresis (PFGE), Multi-Locus Sequence Typing (MLST) and spa typing. The MLVF analysis showed almost the same clonality results as the remaining typing methods although some differences were found. Epidemiological data about the relation among S. aureus isolates and the results produced by typing methods applied in the present study indicate that because of the advantages in ease and speed of Variable Number of Tandem Repeats (VNTR) procedure over PFGE, spa typing and MLST, MLVF can be used as a first screening method followed by additional typing.     

Primary effect of bromoxynil to induce plant cell death may be cytosol acidification

Bromoxynil, 3,5-dibromo-4-hydroxybenzonitrile, is a commonly used herbicide and is also used as a tool to trigger rapid cell death in basic botany. However, the primary effect inducing cell death is not known. Bromoxynil inhibited the cytoplasmic streaming and killed cells in Chara corallina when it was applied in the acidic external medium. At higher pH, bromoxynil was inert even at high concentrations. It was speculated that bromoxynil in the protonated form enters the cell and acidifies the cytosol by releasing H⁺. Experiments using analogues of bromoxynil supported this possibility. Acidification of the cytosol by bromoxynil was confirmed by experiments using pollen tubes. Based on the acidity of the apoplast, the herbicide action of bromoxynil in higher plants was discussed.     

Optimisation of NMR dynamic models II. A new methodology for the dual optimisation of the model-free parameters and the Brownian rotational diffusion tensor

Finding the dynamics of an entire macromolecule is a complex problem as the model-free parameter values are intricately linked to the Brownian rotational diffusion of the molecule, mathematically through the autocorrelation function of the motion and statistically through model selection. The solution to this problem was formulated using set theory as an element of the universal set —the union of all model-free spaces (d’Auvergne EJ and Gooley PR (2007) Mol BioSyst 3(7), 483–494). The current procedure commonly used to find the universal solution is to initially estimate the diffusion tensor parameters, to optimise the model-free parameters of numerous models, and then to choose the best model via model selection. The global model is then optimised and the procedure repeated until convergence. In this paper a new methodology is presented which takes a different approach to this diffusion seeded model-free paradigm. Rather than starting with the diffusion tensor this iterative protocol begins by optimising the model-free parameters in the absence of any global model parameters, selecting between all the model-free models, and finally optimising the diffusion tensor. The new model-free optimisation protocol will be validated using synthetic data from Schurr JM et al. (1994) J Magn Reson B 105(3), 211–224 and the relaxation data of the bacteriorhodopsin (1–36)BR fragment from Orekhov VY (1999) J Biomol NMR 14(4), 345–356. To demonstrate the importance of this new procedure the NMR relaxation data of the Olfactory Marker Protein (OMP) of Gitti R et al. (2005) Biochem 44(28), 9673–9679 is reanalysed. The result is that the dynamics for certain secondary structural elements is very different from those originally reported. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Expression pattern of <Emphasis Type="Italic">serine protease inhibitor kazal type 3 (Spink3)</Emphasis> during mouse embryonic development

Recent evidence shows that the serine protease inhibitor Kazal type 3 (Spink3) has more diverse functions than expected. To gain insight into its function, we analyzed the spatiotemporal expression profile of Spink3, using in situ hybridization (ISH) and a Spink3 ( +/lacZ ) knock-in mouse, in which lacZ was inserted into the Spink3 locus. Spink3 ( lacZ ) expression was first observed in the foregut, midgut, hindgut and the forebrain/midbrain junction region at 9.5 days post coitus (dpc). In the pancreas, Spink3 mRNA was detected at 11.5 dpc, before formation of the typical shape of the exocrine structure of the pancreas. Acinar cell expression was clearly identified by 13.5 dpc. After differentiation of the intestinal tract, Spink3 ( lacZ ) expression was observed in the large intestine at 11.5 dpc, followed by expression in the small intestine at 13.5 dpc, before appearance of intestinal digestive enzymes. Spink3 mRNA and Spink3 ( lacZ ) activity were also detected in other tissues, including the mesonephric tubules and the urogenital ridge at 11.5 dpc, the genital swelling at 13.5 dpc, the ductus epididymis at 17.5 dpc, and the seminal vesicle at 8 weeks. These data suggest that Spink3 may play important roles in proliferation and/or differentiation of various cell types during development.