Localization of neogenin protein during morphogenesis in the mouse embryo

Neogenin, a close relative of the axon guidance receptor DCC, has been shown to be a receptor for members of the Netrin and Repulsive Guidance Molecule families. Recent studies have begun to uncover a role for Neogenin in organogenesis. Here we examine the localization of Neogenin protein in the developing mouse embryo (embryonic day 14.5) when organogenesis is progressing rapidly. We observe that Neogenin protein is restricted to distinct tissue layers within a given organ. In some embryonic epithelia such as the gut and pancreas, Neogenin protein is predominantly polarized to the basal surfaces of the epithelial cells. In contrast, Neogenin is restricted to mesenchymal cells within the lung and kidney. Neogenin is also seen in differentiating skeletal muscle and condensing cartilage throughout the embryo, and in the trigeminal and dorsal root ganglia of the peripheral nervous system. This study supports the emerging role for Neogenin as a key receptor in the establishment of the morphological architecture in many developing organ systems.

Dilution versus dialysis - A quantitative study of the oxidative refolding of recombinant human alpha-fetoprotein

Alpha-fetoprotein (AFP) is a commercially important polypeptide with important diagnostic. physiological and immunomodulatory functions. Previous studies into the refolding of this macromolecule are contradictory. and variously suggest that AFP denaturation may be irreversible or that refolding may be achieved by reducing denaturant concentration through dilution but not dialysis. Importantly, these same previous studies do not provide quantitative metrics by which the Success of refolding, and the potential for bioprocess development. can be assessed. Moreover, these same studies do not optimize and control refolding redox potential - an important factor considering that AFP contains 32 cysteines which form 16 disulfide bonds. In this current study, a quantitative comparison of recombinant human AFP (rhAFP) refolding by dilution and dialysis is conducted under optimized redox conditions. rhAFP refolding yields were > 35% (dialysis refolding) and > 75% (dilution refolding) as assessed by RP-HPLC and ELISA, with structural Similarity to the native state confirmed by UV spectroscopy. Dialysis refolding yield was believed to be lower because the gradual reduction in denaturant concentration allowed extended conformational searching. enabling more time for undesirable interaction with other protein molecules and/or the dialysis membrane, leading to a Sub-optimal process outcome. Significant yield sensitivity to redox environment was also observed, emphasizing the importance of physicochemical optimization. This study demonstrates that very high refolding yields can be obtained, for a physiologically relevant protein, with optimized dilution refolding. The study also highlights the quantitative metrics and macromolecular physical spectroscopic 'fingerprints' required to facilitate transition from laboratory to process scale.

The renal cortical fibroblast in renal tubulointerstitial fibrosis

Renal cortical fibroblasts have key roles in mediating intercellular communication with neighboring/infiltrating cells and extracellular matrix (ECM) and maintenance of renal tissue architecture. They express a variety of cytokines, chemokines, growth factors and cell adhesion molecules, playing an active role in paracrine and autocrine interactions and regulating both fibrogenesis and the interstitial inflammatory response. They additionally have an endocrine function in the production of epoetin. Tubulointerstitial fibrosis, the common pathological consequence of renal injury, is characterized by the accumulation of extracellular matrix largely due to excessive production in parallel with reduced degradation, and activated fibroblasts characterized by a myofibroblastic phenotype. Fibroblasts in the kidney may derive from resident fibroblasts, from the circulating fibroblast population or from haemopoetic progenitor or stromal cells derived from the bone marrow. Cells exhibiting a myofibroblastic phenotype may derive from these sources and from tubular cells undergoing epithelial to mesenchymal transformation in response to renal injury. The number of interstitial myofibroblasts correlates closely with tubulointerstitial fibrosis and progressive renal failure. Hence inhibiting myofibroblast formation may be an effective strategy in attenuating the development of renal failure in kidney disease of diverse etiology. (c) 2005 Elsevier Ltd. All rights reserved.

Ireland and immigration: explaining the absence of the far right

Garner seeks to explain the absence of far-right political formations in the history of the Republic of Ireland, especially in relation to immigration. He argues that the ‘mainstream’ nationalist parties have implemented a racialized governance of Ireland via the issue of citizenship (in the referendum of 2004). While hegemonic ideas on the racial purity of indigenous populations and the highly ambivalent attitudes and policies on immigration pursued over the last decade are characteristic of a broader European trend, this has not, in the Republic, been accompanied by meaningful far-right political mobilization. Ireland has frequently been seen as sui generis in political terms, and indeed emerges in some ways as a counter-case: increasing hostility towards Others has been identified in the midst of rapid economic growth and political stability. A variety of issues related to the country’s political development have given rise to an especially small left-wing vote, a nationalist centre ground and longlasting domination by a single populist party, Fianna Fa´ il. This party has been partnered in government since 1997 by a free-market party, the Progressive Democrats, who have contributed to Ireland’s movement towards neo-liberal policies and a highly functional approach to immigration. The transition from country of emigration to country of immigration has thus taken place against an ideological backdrop in which the imperatives of labour demand and consolidating domestic support for reform have made an uneasy match, resulting in the racialization of Irishness. The state has, however, amended the Constitution in order to qualify jus soli citizenship entitlement in the case of particular categories of people: those whose parents are not Irish nationals. The significant stakes of these changes are analysed in the context of state responses to Eire’s transition to a country of immigration, and the role of nationalist-populism in the country’s political culture.

Minimizing unsatisfaction in colourful neighbourhoods

Colouring sparse graphs under various restrictions is a theoretical problem of significant practical relevance. Here we consider the problem of maximizing the number of different colours available at the nodes and their neighbourhoods, given a predetermined number of colours. In the analytical framework of a tree approximation, carried out at both zero and finite temperatures, solutions obtained by population dynamics give rise to estimates of the threshold connectivity for the incomplete to complete transition, which are consistent with those of existing algorithms. The nature of the transition as well as the validity of the tree approximation are investigated.

Dielectric and electro-optical properties of some cyanobiphenyl liquid-crystals

A variety of methods have been reviewed for obtaining parallel or perpendicular alignment in liquid-crystal cells. Some of these methods have been selected and developed and were used in polarised spectroscopy, dielectric and electro-optic studies. Also, novel dielectric and electro-optic cells were constructed for use over a range of temperature. Dielectric response of thin layers of E7 and E8 (eutectic mixture liquid-crystals) have been measured in the frequency range (12 Hz-100 kHz) and over a range of temperature (183-337K). Dielectric spectra were also obtained for supercooled E7 and E8 in the Hz and kHz range. When the measuring electric field was parallel to the nematic director, one loss peak (low-frequency relaxation process) was observed for E7 and for E8, that exhibits a Debye-type behaviour in the supercooled systems. When the measuring electric field was perpendicular to the nematic director, two resolved dielectric processes have been observed. The phase transitions, effective molecular polarisabilities, anisotropy of polarisabilities and order parameters of three liquid crystal homologs (5CB, 6CB, and 7CB), 60CB and three eutectic nematic mixtures E7, E8, and E607 were calculated using optical and density data measured at several temperatures. The order parameters calculated using the different methods of Vuks, Neugebauer, Saupe-Maier, and Palffy-Muhoray are nearly the same for the liquid crystals considered in the present study. Also, the interrelationship between density and refractive index and the molecular structure of these liquid crystals were established. Accurate dielectric and dipole results of a range of liquid-crystal forming molecules at several temperatures have reported. The role of the cyano-end group, biphenyl core, and flexible tail in molecular association, were investigated using the dielectric method for some molecules which have a structural relationship to the nematogens. Analysis of the dielectric data for solution of the liquid-crystals indicated a high molecular association, comparable to that observed in the nematic or isotropic phases. Electro-optic Kerr effect were investigated for some alkyl cyanobiphenyls, their nematic mixtures and the eutectic mixture liquid-crystals E7 and E8 in the isotropic phase and solution. The Kerr constant of these liquid crystals found to be very high at the nematic-isotropic transition temperatures as the molecules are expected to be highly ordered close to phase transition temperatures. Dynamic Kerr effect behaviour and transient molecular reorientation were also observed in thin layers of some alkyl cyanobiphenyls. Dichroic ratio R and order parameters of solutions containing some azo and anthraquinone dyes in the nematic solvent (E7 and E8), were investigated by the measurement of the intensity of the absorption bands in the visible region of parallel aligned samples. The effective factors on the dichroic ratio of the dyes dissolved in the nematic solvents were determined and discussed.

The effect of flow regimes in the distillation of systems containing two liquid phases

Single phase solutions containing three components have been observed to exhibit foaminess near a single to two liquid phase boundary. It was seen, in a sintered plate column under mass transfer conditions, that distillation systems where the liquid appeared as one phase in one part of a column and two phases in another part, exhibited foaminess when the liquid concentration was near the one phase to two phase boundary. Various ternary systems have been studied in a 50 plate. 30mm i.d. Oldershaw column and it was observed that severe foaming occurred in the middle section of the column near the one liquid phase to two liquid phase boundary and no foaming occurred at the end of the column where liquid was either one phase or two phase. This is known as Ross type foam. Mass transfer experiments with Ross type ternary systems have been carried out in a perspex simulator with small and large hole diameter trays. It was observed that by removal of the more volatile component, Ross type foam did not build up on the tray. Severe entrainment of liquid was observed in all cases leading to a 'dry' tray, even with a low free area small diameter hole tray which was expected to produce a bubbly mixture. Entrainment was more severe for high gas superficial velocities and large hole diameters. This behaviour is quite different from the build up of foam observed when one liquid phase/two liquid phase Ross systems were contacted with air above a small sintered disc or with vapour in an Oldershaw distillation column. This observation explains why distillation columns processing mixtures which change from one liquid phase to two liquid phases (or vice versa) must be severely derated to avoid flooding. Single liquid phase holdups at the spray to bubbly transition were measured using a perspex simulator similar to that of Porter & Wong (17). i.e. with no liquid cross flow. A light transmission technique was used to measure the transition from spray regime to bubbly regime. The effect of tray thickness and the ratio of hole diameter to tray thickness on the transition was evaluated using trays of the same hole diameter and free area but having thickness of 2.38 mm, 4 mm, and 6.35 mm. The liquid holdup at the transition was less with the thin metal trays. This result may be interpreted by the theory of Lockett (101), which predicts the transition liquid holdup in terms of the angle of the gas iet leaving the holes in the sieve plate. All the existing correlations have been compared and none were found to be satisfactory and these correlations have been modified in view of the experimental results obtained. A new correlation has been proposed which takes into account the effect of the hole diameter to tray thickness ratio on the transition and good agreement was obtained between the experimental results and the correlated values of the liquid holdup at the transition. Results have been obtained for two immiscible liquids [kerosene and water] on trays to determine whether foaming can be eliminated by operating in the spray regime. Kerosene was added to a fixed volume of water or water was added to a fixed volume of kerosene. In both cases, there was a transition from spray to bubbly. In the water fixed system. the liquid holdup at the transition was slightly less than the pure kerosene system. Whilst for the kerosene fixed system, the transition occurred at much lower liquid holdups. Trends In the results were similar to those for single liquid phase. New correlations have been proposed for the two cases. It has been found that Ross type foams, observed in a sintered plate column and in the Oldershaw column can be eliminated by either carrying out the separation in a packed column or by the addition of defoaming additives.

A study of the calcium-induced morphological transistions of human erythrocytes

An examination was made of the morphological transitions induced in human erythrocytes by the elevation of cytosolic calcium, and of the biochemical mechanisms responsible. The loss of the discocyte morphology and the sequential progression of cells through the echinocyte stages 1, 2, 3 and sphereo-echinocyte was found to occur in both a calcium concentration- and a time-dependent manner. SDS-PAGE analysis of cytoskeletal proteins prepared from intact cells loaded with 150uM or 1mM calcium revealed the partial proteolytic loss of proteins 2.1, 2.2 and 4.1. The rate of proteolysis was not paralleled by that of echinocytosis, making a causative relationship unlikely. Cytoskeletal integrity did appear to influence shape reversal from the echinocyte to the discocyte morphology after removal of the calcium and ionophore A23187. The loss of 80% protein 4.1, 40% 2.1 and 30% 2.2 was associated with, although not necessarily the sole cause, of irreversible sphereo-echinocytosis. Pre-treatment of cells with wheat germ agglutinin preserved the discocyte morphology despite continued cytoskeletal proteolysis during calcium-loading. All observations were made on cells incubated either in the presence or absence of glycolytic substrates, effectively altering cell metabolic status. This influenced the rate of progression of cells through the echinocyte stages, the rate of proteolysis of cytoskeletal proteins, and the extent and kinetics of shape reversal from cells transformed to the sphereo-echinocyte morphology. The stage 1 to discocyte transition was the rate limiting step of this shape recovery. In contrast the rate of loss of the discocyte morphology was independent of cell metabolic status during exposure to calcium, as was the extent of restoration of the discocyte morphology from cells transformed to stage 1 echinocytes. An hypothesis is presented that echinocytosis is a discontinuous process with discrete steps initiated by different biochemical mechanisms varying in their dependence on metabolic energy.

A method for the in-situ determination of the hydraulic conductivity of gravels as used in constructed wetlands for wastewater treatment

A new instrument and method are described that allow the hydraulic conductivities of highly permeable porous materials, such as gravels in constructed wetlands, to be determined in the field. The instrument consists of a Mariotte siphon and a submersible permeameter cell with manometer take-off tubes, to recreate in-situ the constant head permeameter test typically used with excavated samples. It allows permeability to be measured at different depths and positions over the wetland. Repeatability obtained at fixed positions was good (normalised standard deviation of 1–4%), and results obtained for highly homogenous silica sand compared well when the sand was retested in a lab permeameter (0.32 mm.s–1 and 0.31 mm.s–1 respectively). Practical results have a ±30% associated degree of uncertainty because of the mixed effect of natural variation in gravel core profiles, and interstitial clogging disruption during insertion of the tube into the gravel. This error is small, however, compared to the orders of magnitude spatial variations detected. The technique was used to survey the hydraulic conductivity profile of two constructed wetlands in the UK, aged 1 and 15 years respectively. Measured values were high (up to 900 mm.s –1) and varied by three orders of magnitude, reflecting the immaturity of the wetland. Detailed profiling of the younger system suggested the existence of preferential flow paths at a depth of 200 mm, corresponding to the transition between more coarse and less coarse gravel layers (6–12 mm and 3–6 mm respectively), and transverse drift towards the outlet.

Nanophotonics of optical fibers

This review is concerned with nanoscale effects in highly transparent dielectric photonic structures fabricated from optical fibers. In contrast to those in plasmonics, these structures do not contain metal particles, wires, or films with nanoscale dimensions. Nevertheless, a nanoscale perturbation of the fiber radius can significantly alter their performance. This paper consists of three parts. The first part considers propagation of light in thin optical fibers (microfibers) having the radius of the order of 100 nanometers to 1 micron. The fundamental mode propagating along a microfiber has an evanescent field which may be strongly expanded into the external area. Then, the cross-sectional dimensions of the mode and transmission losses are very sensitive to small variations of the microfiber radius. Under certain conditions, a change of just a few nanometers in the microfiber radius can significantly affect its transmission characteristics and, in particular, lead to the transition from the waveguiding to non-waveguiding regime. The second part of the review considers slow propagation of whispering gallery modes in fibers having the radius of the order of 10–100 microns. The propagation of these modes along the fiber axis is so slow that they can be governed by extremely small nanoscale changes of the optical fiber radius. This phenomenon is exploited in SNAP (surface nanoscale axial photonics), a new platform for fabrication of miniature super-low-loss photonic integrated circuits with unprecedented sub-angstrom precision. The SNAP theory and applications are overviewed. The third part of this review describes methods of characterization of the radius variation of microfibers and regular optical fibers with sub-nanometer precision.

Effect of water on olivine single crystals plasticity, deformed under high pressure and high temperature

The Earth's upper mantle, mainly composed of olivine, is seismically anisotropic. Seismic anisotropy attenuation has been observed at 220km depth. Karato et al. (1992) attributed this attenuation to a transition between two deformation mechanisms, from dislocation creep above 220km to diffusion creep below 220km, induced by a change in water content. Couvy (2005) and Mainprice et al. (2005) predicted a change in Lattice Preferred Orientation induced by pressure, which comes from a change of slip system, from [100] slip to [001] slip, and is responsible for the seismic anisotropy attenuation. Raterron et al. (2007) ran single crystal deformation experiments under anhydrous conditions and observed that the slip system transition occurs around 8GPa, which corresponds to a depth of 260Km. Experiments were done to quantify the effects of water on olivine single crystals deformed using D-DIA press and synchrotron beam. Deformations were carried out in uniaxial compression along [110]c, [011]c, and [101]c, crystallographic directions, at pressure ranging from 4 to 8GPa and temperature between 1373 and 1473K. Talc sleeves about the annulus of the single crystals were used as source of water in the assembly. Stress and specimen strain rates were calculated by in-situ X-ray diffraction and time resolved imaging, respectively. By direct comparison of single crystals strain rates, we observed that [110]c deforms faster than [011]c below 5GPa. However above 6GPa [011]c deforms faster than [110]c. This revealed that [100](010) is the dominant slip system below 5GPa, and above 6GPa [001](010) becomes dominant. According to our results, the slip system transition, which is induced by pressure, occurs at 6GPa. Water influences the pressure where the switch over occurs, by lowering the transition pressure. The pressure effect on the slip systems activity has been quantified and the hydrolytic weakening has also been estimated for both orientations. Data also shows that temperature affects the slip system activity. The regional variation of the depth for the seismic anisotropy attenuation, which would depend on local hydroxyl content and temperature variations and explains the seismic anisotropy attenuation occurring at about 220Km depth in the mantle, where the pressure is about 6GPa. Deformation of MgO single crystal oriented [100], [110] and [111] were also performed. The results predict a change in the slip system activity at 23GPa, again induced by pressure. This explains the seismic anisotropy observed in the lower mantle.

The Chef In Society: Origins And Development

In his discourse - The Chef In Society: Origins And Development - Marcel R. Escoffier, Graduate Student, School of Hospitality Management at Florida International University, initially offers: “The role of the modern professional chef has its origins in ancient Greece. The author traces that history and looks at the evolution of the executive chef as a manager and administrator.” “Chefs, as tradespersons, can trace their origins to ancient Greece,” the author offers with citation. “Most were slaves…” he also informs you. Even at that low estate in life, the chef was master of the slaves and servants who were at close hand in the environment in which they worked. “In Athens, a cook was the master of all the household slaves…” says Escoffier. As Athenian influence wanes and Roman civilization picks-up the torch, chefs maintain and increase their status as important tradesmen in society. “Here the first professional societies of cooks were formed, almost a hierarchy,” Escoffier again cites the information. “It was in Rome that cooks established their first academy: Colleqium Coquorum,” he further reports. Chefs, again, increase their significance during the following Italian Renaissance as the scope of their influence widens. “…it is an historical fact that the marriage of Henry IV and Catherine de Medici introduced France to the culinary wonders of the Italian Renaissance,” Escoffier enlightens you. “Certainly the professional chef in France became more sophisticated and more highly regarded by society after the introduction of the Italian cooking concepts.” The author wants you to know that by this time cookbooks are already making important inroads and contributing to the history of cooking above and beyond their obvious informational status. Outside of the apparent European influences in cooking, Escoffier also ephemerally mentions the development of Chinese and Indian chefs. “It is interesting to note that the Chinese, held by at least one theory as the progenitors of most of the culinary heritage, never developed a high esteem for the position of chef,” Escoffier maintains the historical tack. “It was not until the middle 18th Century that the first professional chef went public. Until that time, only the great houses of the nobility could afford to maintain a chef,” Escoffier notes. This private-to-public transition, in conjunction with culinary writing are benchmarks for the profession. Chefs now establish authority and eminence. The remainder of the article devotes itself to the development of the professional chef; especially the melding of two seminal figures in the culinary arts, Cesar Ritz and August Escoffier. The works of Frederick Taylor are also highlighted.

A decision support framework for infrastructure maintenance investment decision-making

Infrastructure management agencies are facing multiple challenges, including aging infrastructure, reduction in capacity of existing infrastructure, and availability of limited funds. Therefore, decision makers are required to think innovatively and develop inventive ways of using available funds. Maintenance investment decisions are generally made based on physical condition only. It is important to understand that spending money on public infrastructure is synonymous with spending money on people themselves. This also requires consideration of decision parameters, in addition to physical condition, such as strategic importance, socioeconomic contribution and infrastructure utilization. Consideration of multiple decision parameters for infrastructure maintenance investments can be beneficial in case of limited funding. Given this motivation, this dissertation presents a prototype decision support framework to evaluate trade-off, among competing infrastructures, that are candidates for infrastructure maintenance, repair and rehabilitation investments. Decision parameters' performances measured through various factors are combined to determine the integrated state of an infrastructure using Multi-Attribute Utility Theory (MAUT). The integrated state, cost and benefit estimates of probable maintenance actions are utilized alongside expert opinion to develop transition probability and reward matrices for each probable maintenance action for a particular candidate infrastructure. These matrices are then used as an input to the Markov Decision Process (MDP) for the finite-stage dynamic programming model to perform project (candidate)-level analysis to determine optimized maintenance strategies based on reward maximization. The outcomes of project (candidate)-level analysis are then utilized to perform network-level analysis taking the portfolio management approach to determine a suitable portfolio under budgetary constraints. The major decision support outcomes of the prototype framework include performance trend curves, decision logic maps, and a network-level maintenance investment plan for the upcoming years. The framework has been implemented with a set of bridges considered as a network with the assistance of the Pima County DOT, AZ. It is expected that the concept of this prototype framework can help infrastructure management agencies better manage their available funds for maintenance.

Beryllium concentrations and deposition rates in ODP holes from the north Atlantic

Increases in the production rate of cosmogenic radionuclides associated with geomagnetic excursions have been used as global tie-points for correlation between records of past climate from marine and terrestrial archives. We have investigated the relative timing of variations in 10Be production rate and the corresponding palaeomagnetic signal during one of the largest Pleistocene excursions, the Iceland Basin (IB) event (ca. 190 kyr), as recorded in two marine sediment cores (ODP Sites 1063 and 983) with high sedimentation rates. Variations in 10Be production rate during the excursion were estimated by use of 230Thxs normalized 10Be deposition rates and authigenic 10Be/9Be. Resulting 10Be production rates are compared with high-resolution records of geomagnetic field behaviour acquired from the same discrete samples. We find no evidence for a significant lock-in depth of the palaeomagnetic signal in these high sedimentation-rate cores. Apparent lock-in depths in other cores may sometimes be the result of lower sample resolution. Our results also indicate that the period of increased 10Be production during the IB excursion lasted longer and, most likely, started earlier than the corresponding palaeomagnetic anomaly, in accordance with previous observations that polarity transitions occur after periods of reduced geomagnetic field intensity prior to the transition. The lack of evidence in this study for a significant palaeomagnetic lock-in depth suggests that there is no systematic offset between the 10Be signal and palaeomagnetic anomalies associated with excursions and reversals, with significance for the global correlation of climate records from different archives.