Origin and evolution of the northern hemisphere disjunction in the moss genus Homalothecium (Brachytheciaceae)

Competing hypotheses that rely either on a stepping-stone dispersal via the North Atlantic or the Bering land bridges, or more recent transoceanic dispersal, have been proposed to explain the disjunct distribution of Mediterranean flora in southern Europe and western North America. These hypotheses were tested with molecular dating using a phylogeny of the moss genus Homalothecium based on ITS, atpB-rbcL, and rpl16 sequence data. The monophyly of two main lineages in Western Palearctic (Europe, central Asia and north Africa) and North America is consistent with the ancient vicariance hypothesis. The monophyly of Madeiran H. sericeum accessions supports the recognition of the Macaronesian endemic H. mandonii. A range of absolute rates of molecular evolution documented in land plants was used as probabilistic calibration prior by a Bayesian inference implementing a relaxed-clock model to derive ages for the nodes of interest. Our age estimates for the divergence of the American and Western Palearctic Homalothecium clade (5.7 Ma, IC 3.52-8.26) and the origin of H. mandonii (2.52 Myr IC 0.86-8.25) are not compatible with the ancient vicariance hypothesis. Age estimates suggests that species distributions result from rare instances of dispersal and subsequent sympatric diversification. The calibrated phylogeny indicates that Homalothecium has undergone a fast radiation during the last 4 Myr, which is consistent with the low levels of morphological divergence among sibling species.     

The effects of shredding invertebrates on the transfer of organic carbon from littoral leaf litter to water-column bacteria

Leaf litter can be of great importance for the productivity of small oligotrophic lakes surrounded by deciduous forests. Feeding invertebrate shredders produce particulate organic leftovers, but their feeding also enhances the release of dissolved organic carbon (DOC). We tested whether invertebrate-mediated DOC release affects the production of heterotrophic water-column bacteria. Submersed leaves were incubated in microcosms with and without shredders; and DOC, absorbance, bacterial abundance and bacterial production in the water column were monitored. We also measured dry weight of the organic particles (FPOC, fine particulate organic carbon, leaf residues and shredders). Total leaf-litter carbon decreased by nearly 80% in the presence of shredders, and on average 56% of the initial leaf carbon ended up as FPOC after 126 days of incubation. Without shredders FPOC production was almost zero, and 72% of the added leaf carbon could be retrieved as leaves when the experiment ended. Both these figures include the rapid release of DOC during the first week of leaf incubation in the lake water (equivalent to 16–19% of total added leaf carbon). Although bacterial production in the water was several times higher in treatments with shredders, bacterial consumption of leaf-derived DOC from shredding was obviously of minor importance in the total carbon budget. This result suggests, although shredders have a strong impact on transformation of leaves to FPOC, they do not greatly enhance the initial rate of mineralization of the leaf-derived detritus.     

Effects of hunger level and nutrient balance on survival and acetylcholinesterase activity of dimethoate exposed wolf spiders

The influence of two nutritional factors (food quantity and quality) on the responses of a wolf spider, Pardosa prativaga (L.K.), to a high dose of the insecticide dimethoate, was investigated in a fully factorial experimental design. Spider groups with different (good and bad) nutrient balance were created by feeding them fruit flies of either high or low nutrient content for 28 days. Both groups were then split into satiated and 14 days starved subgroups. Each of these was further divided into insecticide treated and control halves. Survivorship and acetylcholinesterase (AChE) activity measured on the survivors were used as response variables. Survivorship after topical dimethoate exposure (LD₅₀; 48 h) was influenced by spider body weight, nutrient balance, and starvation. Furthermore, AChE activity was significantly inhibited by dimethoate exposure. A significant interaction between nutrient balance, starvation, and dimethoate exposure revealed synergistic effects of starvation and nutrient imbalance on AChE inhibition by dimethoate in surviving spiders. These results show that the tolerance of non-target arthropods to dimethoate may vary depending on the nutritional history of the animal.     

Expression of recombinant murine pregnancy-associated plasma protein-A (PAPP-A) and a novel variant (PAPP-Ai) with differential proteolytic activity.

Murine pregnancy-associated plasma protein-A (PAPP-A) cDNA encoding a 1545 amino-acid protein has been cloned. We have also identified and cloned cDNA that encodes a novel variant of PAPP-A, PAPP-Ai, carrying a 29-residue highly basic insert. The point of insertion corresponds to a junction between two exons in the human PAPP-A gene. The human intron flanked by these exons does not encode a homologous corresponding insert, which is unique to the mouse. The overall sequence identity between murine and human PAPP-A is 91%, and murine PAPP-A contains sequence motifs previously described in the sequence of human PAPP-A. Through expression in mammalian cells, we show that murine PAPP-A and PAPP-Ai are active metalloproteinases, both capable of cleaving insulin-like growth factor binding protein (IGFBP)-4 and -5. Cleavage of IGFBP-4 is dramatically enhanced by the addition of IGF, whereas cleavage of IGFBP-5 is slightly inhibited by IGF, as previously established with human PAPP-A. Surprisingly, however, quantitative analyses demonstrate that the murine PAPP-Ai cleaves IGFBP-4 very slowly compared to PAPP-A, even though its ability to cleave IGFBP-5 is unaffected by the presence of the insert. By RT-PCR analysis, we find that both variants are expressed in several tissues. The level of mRNA in the murine placenta does not exceed the levels of other tissues analyzed. Furthermore, the IGFBP-4-proteolytic activity of murine pregnancy serum is not elevated. This is in striking contrast to the increase seen in human pregnancy serum, and the expression of PAPP-A in the human placenta, which exceeds other tissues at least 250-fold. Interestingly, the position of the insert of PAPP-Ai, within the proteolytic domain, lies in close proximity to the cysteine residue, which in human PAPP-A forms a disulfide bond with the proform of eosinophil major basic protein (proMBP). ProMBP functions as a proteinase inhibitor in the PAPP-A-proMBP complex, but whether any mechanistic parallel on regulation of proteolytic activity can be drawn between the insert of PAPP-Ai and the linkage to proMBP is not known. Importantly, these data support the development of the mouse as a model organism for the study of PAPP-A, which must take into account the differences between the mouse and the human.     

Comparative analysis of cobalamin binding kinetics and ligand protection for intrinsic factor, transcobalamin, and haptocorrin.

Changes in the absorbance spectrum of aquo-cobalamin (Cbl x OH(2)) revealed that its binding to transcobalamin (TC) is followed by slow conformational reorganization of the protein-ligand complex (Fedosov, S. N., Fedosova, N. U., Nex?, E., and Petersen, T. E. (2000) J. Biol. Chem. 275, 11791-11798). Two phases were also observed for TC when interacting with a Cbl-analogue cobinamide (Cbi), but not with other cobalamins. The slow phase had no relation to the ligand recognition, since both Cbl and Cbi bound rapidly and in one step to intrinsic factor (IF) and haptocorrin (HC), namely the proteins with different Cbl specificity. Spectral transformations observed for TC in the slow phase were similar to those upon histidine complexation with Cbl x OH(2) and Cbi. In contrast to a closed structure of TC x Cbl x OH(2), the analogous IF and HC complexes revealed accessibility of Cbl's upper face to the external reagents. The binders decreased sensitivity of adenosyl-Cbl (Cbl x Ado) to light in the range: free ligand, IF x, HC x, TC x Cbl x Ado. The spectrum of TC x Cbl small middle dotAdo differed from those of IF and HC and mimicked Cbl x Ado participating in catalysis. The above data suggest presence of a histidine-containing cap shielding the Cbl-binding site in TC. The cap coordinates to certain corrinoids and, possibly, produces an incapsulated Ado-radical when Cbl small middle dotAdo is bound.     

Distribution of class I, III and IV alcohol dehydrogenase mRNAs in the adult rat, mouse and human brain.

The localization of different classes of alcohol dehydrogenases (ADH) in the brain is of great interest because of their role in both ethanol and retinoic acid metabolism. Conflicting data have been reported in the literature. By Northern blot and enzyme activity analyses only class III ADH has been detected in adult brain specimens, while results from riboprobe in situ hybridization indicate class I as well as class IV ADH expression in different regions of the rat brain. Here we have studied the expression patterns of three ADH classes in adult rat, mouse and human tissues using radioactive oligonucleotide in situ hybridization. Specificity of probes was tested on liver and stomach control tissue, as well as tissue from class IV ADH knock-out mice. Only class III ADH mRNA was found to be expressed in brain tissue of all three investigated species. Particularly high expression levels were found in neurons of the red nucleus in human tissue, while cortical neurons, pyramidal and granule cells of the hippocampus and dopamine neurons of substantia nigra showed moderate expression levels. Purkinje cells of cerebellum were positive for class III ADH mRNA in all species investigated, whereas granular layer neurons were positive only in rodents. The choroid plexus was highly positive for class III ADH, while no specific signal for class I or class IV ADH was detected. Our results thus support the notion that the only ADH expressed in adult mouse, rat and human brain is class III ADH.     

The brachiopod fold: a neglected body plan hypothesis

Attention is drawn to Nielsen's radical body plan concept, here named the 'brachiopod fold hypothesis', under which brachiopods and phoronids are recognized to be transversely folded across the ontogenetic anterior–posterior axis so that, to make useful comparisons with other phyla, these organisms must be conceptually unfolded. Under the hypothesis brachiopod brachial and pedicle shell valves are respectively 'anterior' and 'posterior' rather than 'dorsal' and 'ventral' as traditionally described. The hypothesis makes sense of the symmetry axes of the brachiopod shell, is consistent with various indications from fossil and Recent brachiopods, and gives rise to predicted patterns of axis–determining gene expression that differ from those obtaining under the traditional view of the body plan, whilst the variety of folding movements in different lineages implies that superficially dissimilar morphogenetic folds may be fundamentally homologous. Convergent folding patterns are noted in some other organisms. A previous conjecture that inarticulate linguloid brachiopods were derived from halkieriid–like ancestors is elaborated with proposals that recognize possible functional continuities of coelomic and marginal sclerite functions, and it is noted that an ancestrally facultative fold could have become incorporated by genetic assimilation into the brachiopod developmental program. An experimental approach is outlined to test the possibility that some members of the 'small shelly fauna' may have been members of the halkieriid–like brachiopod stem lineage and it is also suggested that buoyancy modification may have been an important function of mineralization amongst Lower Cambrian floaters and swimmers, since negative buoyancy would facilitate access to the benthic niche.     

Evolution, mitochondrial DNA phylogeny and systematic position of the Macaronesian endemic Hydrotarsus Falkenström (Coleoptera: Dytiscidae)

Abstract.  Hydrotarsus Falkenström (Coleoptera, Dytiscidae) is the only genus of aquatic beetle considered to be endemic to the Atlantic islands of Macaronesia. Its three known species ( H. lundbladi Falkenström from Madeira, and H. compunctus (Wollaston) and H. pilosus Guignot from the Canary Islands) are revised, and their phylogenetic position studied based on sequences of the 16S rRNA and Cytochrome Oxidase I genes of mitochondrial DNA. Our results clearly indicate that the species of Hydrotarsus fall deep within the genus Hydroporus Clairville, in a clade formed by (in addition to Hydrotarsus ) species currently included in the H. fuscipennis , marginatus , nigrita and tessellatus groups, and in consequence a new synonymy is proposed: Hydrotarsus Falkenström =  Hydroporus Clairville syn.n. The origin of the species of ' Hydrotarsus ', based on molecular clock estimations, is late Miocene, relatively recent in the history of the Atlantic islands. They are therefore not palaeoendemics but highly derived, recently evolved elements in the Macaronesian fauna. The estimated ages of divergence among species are much more recent than the emergence of any of the islands on which they are found, suggesting rare long-range dispersal as the mechanism driving the diversification of the lineage. Morphological modifications of the species of ' Hydrotarsus ' are discussed, as well as those of other dytiscids from hygropetric habitats.