Tracing reinforcement through asymmetrical partner preference in the European common vole Microtus arvalis

Background The mechanistic basis of speciation and in particular the contribution of behaviour to the completion of the speciation process is often contentious. Contact zones between related taxa provide a situation where selection against hybridization might reinforce separation by behavioural mechanisms, which could ultimately fully isolate the taxa. One of the most abundant European mammals, the common vole Microtus arvalis, forms multiple natural hybrid zones where rapidly diverging evolutionary lineages meet in secondary contact. Very narrow zones of hybridization spanning only a few kilometres and sex-specific gene flow patterns indicate reduced fitness of natural hybrids and incipient speciation between some of the evolutionary lineages. In this study, we examined the contribution of behavioural mechanisms to the speciation process in these rodents by fine-mapping allopatric and parapatric populations in the hybrid zone between the Western and Central lineages and experimental testing of the partner preferences of wild, pure-bred and hybrid female common voles. Results Genetic analysis based on microsatellite markers revealed the presence of multiple parapatric and largely non-admixed populations at distances of about 10 km at the edge of the area of natural hybridization between the Western and Central lineages. Wild females from Western parapatric populations and lab-born F1 hybrids preferred males from the Western lineage whereas wild females of Central parapatric origin showed no measurable preference. Furthermore, wild and lab-born females from allopatric populations of the Western or Central lineages showed no detectable preference for males from either lineage. Conclusions The detected partner preferences are consistent with asymmetrical reinforcement of pre-mating reproductive isolation mechanisms in the European common vole and with earlier results suggesting that hybridization is more detrimental to the Western lineage. As a consequence, these differences in behaviour might contribute to a further geographical stabilization of this moving hybrid zone. Such behavioural processes could also provide a mechanistic perspective for frequently-detected asymmetrical introgression patterns in the largely allopatrically diversifying Microtus genus and other rapidly speciating rodents.

Computer simulation of electrophoretic aspects of enantiomer migration and separation in capillary electrochromatography with a neutral selector.

A computer simulation study describing the electrophoretic separation and migration of methadone enantiomers in presence of free and immobilized (2-hydroxypropyl)-β-CD is presented. The 1:1 interaction of methadone with the neutral CD was simulated by using experimentally determined mobilities and complexation constants for the complexes in a low-pH BGE comprising phosphoric acid and KOH. The use of complex mobilities represents free solution conditions with the chiral selector being a buffer additive, whereas complex mobilities set to zero provide data that mimic migration and separation with the chiral selector being immobilized, that is CEC conditions in absence of unspecific interaction between analytes and the chiral stationary phase. Simulation data reveal that separations are quicker, electrophoretic displacement rates are reduced, and sensitivity is enhanced in CEC with on-column detection in comparison to free solution conditions. Simulation is used to study electrophoretic analyte behavior at the interface between sample and the CEC column with the chiral selector (analyte stacking) and at the rear end when analytes leave the environment with complexation (analyte destacking). The latter aspect is relevant for off-column analyte detection in CEC and is described here for the first time via the dynamics of migrating analyte zones. Simulation provides insight into means to counteract analyte dilution at the column end via use of a BGE with higher conductivity. Furthermore, the impact of EOF on analyte migration, separation, and detection for configurations with the selector zone being displaced or remaining immobilized under buffer flow is simulated. In all cases, the data reveal that detection should occur within or immediately after the selector zone.

Dielectrophoresis-based analyte separation and analysis

Dielectrophoresis—the tendency of a material of high dielectric permittivity to migrate in an electrical field gradient to a region of maximum field strength—provides an ideal motive force for manipulating small volumes of biological analytes in microfluidic microsystems. The work described in this thesis was based on the hypothesis that dielectrophoresis could be exploited to provide high-resolution cell separations in microsystems as well as a means for the electrically-controllable manipulation of solid supports for molecular analysis. To this end, a dielectrophoretic/gravitational field-flow-fractionation (DEP/G-FFF) system was developed and the separation performance evaluated using various types and sizes of polystyrene microspheres as model particles. It was shown that separation of the polystyrene beads was based on the differences in their effective dielectrophoretic properties. The ability of an improved DEP/G-FFF system to separate genetically identical, but phenotypically dissimilar cell types was demonstrated using mixtures of 6m2 mutant rat kidney cells grown under transforming and non-transforming culture conditions. Additionally, a panel of engineered dielectric microspheres was designed with specific, predetermined dielectrophoretic properties such that their dielectrophoretic behaviors would be controllable and predictable. The fabrication method involved the use of gold-coated polystyrene microsphere cores coated with a self-assembled monolayer of alkanethiol and, optionally, a self-assembled monolayer of phospholipid to form a thin-insulating-shell-over-conductive-interior structure. The successful development of the DEP/G-FFF separation system and the dielectrically engineered microspheres provides proof-of-principle demonstrations of enabling dielectrophoresis-based microsystem technology that should provide powerful new methods for the manipulation, separation and identification of analytes in many diverse fields. ^

Organisation und Separation

Lazar Felix Pinkus

Is maternal separation anxiety associated with school readiness?

Background. Increasing rates of maternal employment highlight the need for non-maternal child care for infants at an earlier age. Several studies have shown that employment induced maternal depression or psychological distress is associated with the child's socio-emotional and cognitive development. However, separation anxiety, a common phenomenon observed among employed mothers during early years, has seldom been studied. Therefore, the purpose of this study was to evaluate the role of maternal separation anxiety in the child's cognitive development.^ Methods. Data were obtained from Phase I (birth to 36 months) of the National Institute of Child Health and Human Development, Study of Early Child Care and Youth Development (NICHD SECCYD). Bivariate and multivariate analyses were performed to determine the association between separation anxiety groups and child outcomes. Multivariate analysis was also used to examine the mediating and/or moderating effect of sensitivity and moderating effect of difficult temperament.^ Results. Separation anxiety showed a negative association with the Bracken, attachment security, maternal sensitivity and psychological state. Children whose mothers never reported high levels of separation anxiety showed higher levels of school readiness and attachment security compared to those whose mothers experienced high levels of separation anxiety at least once. There was a significant interaction between separation anxiety and maternal sensitivity for the Bracken and attachment security indicating the moderating effect of sensitivity. Maternal sensitivity was also found to partially mediate the association between high levels of separation anxiety and school readiness or attachment security. However, the interaction between difficult temperament and separation anxiety was not significant for any of the child outcomes. ^ Conclusions. High levels of separation anxiety have a negative impact on school readiness, attachment security, maternal sensitivity and psychological state. In addition, mothers who experience high levels of separation anxiety but are sensitive during the mother-child interaction have children with high school readiness and attachment security compared to those who are less sensitive.^ Keywords. Maternal separation anxiety, School readiness. ^

Development of combined micro -preparation, separation, and mass spectrometry methods and application in the microdialysis study of neuropeptide metabolism

Two sets of mass spectrometry-based methods were developed specifically for the in vivo study of extracellular neuropeptide biochemistry. First, an integrated micro-concentration/desalting/matrix-addition device was constructed for matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) to achieve attomole sensitivity for microdialysis samples. Second, capillary electrophoresis (CE) was incorporated into the above micro-liquid chromatography (LC) and MALDI MS system to provide two-dimensional separation and identification (i.e. electrophoretic mobility and molecular mass) for the analysis of complex mixtures. The latter technique includes two parts of instrumentation: (1) the coupling of a preconcentration LC column to the inlet of a CE capillary, and (2) the utilization of a matrix-precoated membrane target for continuous CE effluent deposition and for automatic MALDI MS analysis (imaging) of the CE track.^ Initial in vivo data reveals a carboxypeptidase A (CPA) activity in rat brain involved in extracellular neurotensin metabolism. Benzylsuccinic acid, a CPA inhibitor, inhibited neurotensin metabolite NT1-12 formation by 70%, while inhibitors of other major extracellular peptide metabolizing enzymes increased NT1-12 formation. CPA activity has not been observed in previous in vitro experiments. Next, the validity of the methodology was demonstrated in the detection and structural elucidation of an endogenous neuropeptide, (L)VV-hemorphin-7, in rat brain upon ATP stimulation. Finally, the combined micro-LC/CE/MALDI MS was used in the in vivo metabolic study of peptide E, a mu-selective opioid peptide with 25 amino acid residues. Profiles of 88 metabolites were obtained, their identity being determined by their mass-to-charge ratio and electrophoretic mobility. The results indicate that there are several primary cleavage sites in vivo for peptide E in the release of its enkephalin-containing fragments. ^

Flow separation and roughness lengths over large bedforms in a tidal environment

This study characterises the shape of the flow separation zone (FSZ) and wake region over large asymmetric bedforms under tidal flow conditions. High resolution bathymetry, flow velocity and turbulence data were measured along two parallel transects in a tidal channel covered with bedforms. The field data are used to verify the applicability of a numerical model for a systematic study using the Delft3D modelling system and test the model sensitivity to roughness length. Three experiments are then conducted to investigate how the FSZ size and wake extent vary depending on tidally-varying flow conditions, water levels and bathymetry. During the ebb, a large FSZ occurs over the steep lee side of each bedform. During the flood, no flow separation develops over the bedforms having a flat crest; however, a small FSZ is observed over the steepest part of the crest of some bedforms, where the slope is locally up to 15°. Over a given bedform morphology and constant water levels, no FSZ occurs for velocity magnitudes smaller than 0.1 m s**-1; as the flow accelerates, the FSZ reaches a stable size for velocity magnitudes greater than 0.4 m s**-1. The shape of the FSZ is not influenced by changes in water levels. On the other hand, variations in bed morphology, as recorded from the high-resolution bathymetry collected during the tidal cycle, influence the size and position of the FSZ: a FSZ develops only when the maximum lee side slope over a horizontal distance of 5 m is greater than 10°. The height and length of the wake region are related to the length of the FSZ. The total roughness along the transect lines is an order of magnitude larger during the ebb than during the flood due to flow direction in relation to bedform asymmetry: during the ebb, roughness is created by the large bedforms because a FSZ and wake develops over the steep lee side. The results add to the understanding of hydrodynamics of natural bedforms in a tidal environment and may be used to better parameterise small-scale processes in large-scale studies.

Temperature observation of CORK installation during ODP Leg 168

Four boreholes, drilled a few tens of meters into igneous basement on the eastern flank of the Juan de Fuca Ridge during ODP Leg 168, were sealed and instrumented for long-term monitoring to observe the hydrologic state of young sediment-sealed oceanic crust. The thermal regime is dominated by the effects of rapid fluid circulation in uppermost igneous basement driven by very small non-hydrostatic pressure gradients. Upper basement temperatures are uniform laterally between pairs of holes over distances of hundreds of meters to kilometers. In the case of two holes drilled into a sediment-buried basement ridge and adjacent valley, basement temperatures differ by less than 2 K despite the 2.2 km lateral separation of the sites and the 2.5:1 contrast in sediment cover thickness. Under conductive conditions, upper basement temperatures would differ by roughly 50 K. By comparison with modeling results, the observed degree of isothermality suggests a fluid flux of at least 10**-6 m/s (30 m/yr), and an effective permeability in the range of 10**-10 to 10**-9 m**2 in the uppermost igneous crust. The pressure difference available to drive this rapid flux between the ridge and valley, estimated by comparing the observed pressures via the isothermal upper basement hydrostat that is inferred to connect the two sites, is small (~2 kPa) and also suggests high permeability. Relative to the hydrostats defined by the local conductive sediment geotherms, substantial super-hydrostatic pressure (+18 kPa) is present within the buried basement ridge, and sub-hydrostatic pressure is present in the adjacent valley (-26 kPa). Such pressure differentials are the direct consequence of the advection-dominated thermal regime and small pressure losses in high-permeability basement, and are available to drive fluid seepage through sediment sections vertically up above and horizontally away from buried ridges, and down above valleys. No constraints are provided by any of the observations on the depth in the crust to which thermally or chemically significant flow might extend, although just as in the overlying sediments, the pattern of deep flow may be affected by the near-isothermal and near-hydrostatic conditions present in the permeable uppermost crustal section.