Co-ordinating retinal histogenesis: early cell cycle exit enhances early cell fate determination in the Xenopus retina

The laminar arrays of distinct cell types in the vertebrate retina are built by a histogenic process in which cell fate is correlated with birth order. To explore this co-ordination mechanistically, we altered the relative timing of cell cycle exit in the developing Xenopus retina and asked whether this affected the activity of neural determinants. We found that Xath5, a bHLH proneural gene that promotes retinal ganglion cell (RGC) fate, ( Kanekar, S., Perron, M., Dorsky, R., Harris, W. A., Jan, L. Y., Jan, Y. N. and Vetter, M. L. (1997) Neuron 19, 981-994), does not cause these cells to be born prematurely. To drive cells out of the cell cycle early, therefore, we misexpressed the cyclin kinase inhibitor, p27Xic1. We found that early cell cycle exit potentiates the ability of Xath5 to promote RGC fate. Conversely, the cell cycle activator, cyclin E1, which inhibits cell cycle exit, biases Xath5-expressing cells toward later neuronal fates. We found that Notch activation in this system caused cells to exit the cell cycle prematuely, and when it is misexpressed with Xath5, it also potentiates the induction of RGCs. The potentiation is counteracted by co-expression of cyclin E1. These results suggest a model of histogenesis in which the activity of factors that promote early cell cycle exit enhances the activity of factors that promote early cellular fates

Prominin-1 Modulates Rho/ROCK-Mediated Membrane Morphology and Calcium-Dependent Intracellular Chloride Flux

Membrane morphology is an important structural determinant as it reflects cellular functions. The pentaspan membrane protein Prominin-1 (Prom1/CD133) is known to be localised to protrusions and plays a pivotal role in migration and the determination of cellular morphology; however, the underlying mechanism of its action have been elusive. Here, we performed molecular characterisation of Prom1, focussing primarily on its effects on cell morphology. Overexpression of Prom1 in RPE-1 cells triggers multiple, long, cholesterol-enriched fibres, independently of actin and microtubule polymerisation. A five amino acid stretch located at the carboxyl cytosolic region is essential for fibre formation. The small GTPase Rho and its downstream Rho-associated coiled-coil-containing protein kinase (ROCK) are also essential for this process, and active Rho colocalises with Prom1 at the site of initialisation of fibre formation. In mouse embryonic fibroblast (MEF) cells we show that Prom1 is required for chloride ion efflux induced by calcium ion uptake, and demonstrate that fibre formation is closely associated with chloride efflux activity. Collectively, these findings suggest that Prom1 affects cell morphology and contributes to chloride conductance

Miscibility gap of B2 phase in NiAl to Cu3Al section of the Cu-Al-Ni system

The phase separation in the bee phase of the Cu-Al-Ni system at 600-700 degrees C was investigated mainly by energy dispersion X-ray spectrometry (EDS) and differential scanning, calorimetry (DSC). The compositions of the beta(1) (A2 or B2: Cu-rich), beta(2) (B2: NiAl-rich) and gamma (gamma-brass type) phases in equilibrium were determined. It was found that there is a beta(1) + beta(2) miscibility gap in the beta phase region as previously reported by Alexander. It was confirmed by means of high temperature in situ TEM observation that this miscibility gap consists of the B2+B2 phases but not the A2+B2 phases which is sometimes observed in many other Ni-Al and Co-Al base ternary bee alloys. Thermodynamic calculation was performed which indicates that this characteristic feature suggests that the beta(1) (B2) + beta(2) (B2) miscibility gap is a part of a Cu-rich B2 + NiAl-rich B2 miscibility gap island formed around the center of the composition triangle of the isothermal section. The phase separation in the beta phase region and the stability of the ordered bcc alummide are presented and discussed. (c) 2004 Elsevier Ltd. All rights reserved

The management of retinoblastoma

Retinoblastoma (Rb) is the most common primary intraocular malignancy of childhood, but an uncommon paediatric cancer, with a constant incidence worldwide of 1:15,000-1:20,000 live births. Despite its rarity, Rb has served as a cornerstone in the field of oncology in many of the aspects that comprise cancer management, including classification schemes, treatment modalities, genetic testing and screening. Until just over half a century ago, the major treatment for Rb was eye removal, and prognosis was poor with outcome fatal for most children. The dramatic evolution, in a short period of time across all fields of Rb management, as well as the development of specialized centres, better infrastructure and introduction of awareness campaigns, has resulted in nearly 100% survival in developed countries and allowed eye salvage in many of the cases. External beam radiotherapy was used as the main treatment choice for four decades, but replaced by chemotherapy at the turn of the century. Initially, and still in many centres, chemotherapy is administered intravenously, but recently is targeted directly into the eye by means of intra-ophthalmic artery and intravitreal chemotherapy. To date, a range of treatments is available to the Rb expert, including enucleation, but there is lack of consensus in a number of scenarios as to what to use and when. In such a rare cancer, treatment outcomes are reported usually via retrospective analyses, with few prospective randomized controlled trials. Classification schemes have also evolved following the introduction of new treatment modalities, but discrepancies exist among centres with respect to the preferred schema and its interpretation. Retinoblastoma management is a remarkable success story, but the future will require a collaborative effort in the form of multicentre randomized controlled trials in order to further improve the quality of care for this subset of young children with ocular cancer

Experimental determination and thermodynamic calculation of the phase equilibria in the Cu-In-Sn system

The phase equilibria of the Cu-In-Sn system were investigated by means of the diffusion couple method, differential scanning calorimetry (DSC) and metallography. The isothermal sections at 110-900 degreesC, as well as vertical sections at 10wt.%Cu-70wt.%Cu were determined. It was found that there are large solubilities of In in the epsilon (Cu3Sn), delta (Cu41Sn11), and eta phases in the Cu-Sn system, and large solubilities of Sn in the gamma, eta, and delta (Cu7In3) phases in the Cu-In system. The eta phase was found to continuously form from the Cu-In side to the Cu-Sn side, and a ternary compound (Cu2In3Sn) was found to exist at 110 degreesC. Thermodynamic assessment of the Cu-In-Sn system was also carried out based on experimental data of activity and phase equilibria using the CALPHAD method, in which the Gibbs energies of the liquid, fcc and bcc phases are described by the subregular solution model and that of compounds, including two ternary compounds, are represented by the sublattice model. The thermodynamic parameters for describing the phase equilibria were optimized, and agreement between the calculated and experimental results was obtained

Two Secreted Proteoglycans, Activators of Urothelial Cell–Cell Adhesion, Negatively Contribute to Bladder Cancer Initiation and Progression

Osteomodulin (OMD) and proline/arginine-rich end leucine repeat protein (PRELP) are secreted extracellular matrix proteins belonging to the small leucine-rich proteoglycans family. We found that OMD and PRELP were specifically expressed in umbrella cells in bladder epithelia, and their expression levels were dramatically downregulated in all bladder cancers from very early stages and various epithelial cancers. Our in vitro studies including gene expression profiling using bladder cancer cell lines revealed that OMD or PRELP application suppressed the cancer progression by inhibiting TGF-β and EGF pathways, which reversed epithelial–mesenchymal transition (EMT), activated cell–cell adhesion, and inhibited various oncogenic pathways. Furthermore, the overexpression of OMD in bladder cancer cells strongly inhibited the anchorage-independent growth and tumorigenicity in mouse xenograft studies. On the other hand, we found that in the bladder epithelia, the knockout mice of OMD and/or PRELP gene caused partial EMT and a loss of tight junctions of the umbrella cells and resulted in formation of a bladder carcinoma in situ-like structure by spontaneous breakdowns of the umbrella cell layer. Furthermore, the ontological analysis of the expression profiling of an OMD knockout mouse bladder demonstrated very high similarity with those obtained from human bladder cancers. Our data indicate that OMD and PRELP are endogenous inhibitors of cancer initiation and progression by controlling EMT. OMD and/or PRELP may have potential for the treatment of bladder cancer

Phase equilibria of Sn-In based micro-soldering alloys

The phase equilibria of Sn-In-X (X = Ag, Bi, Sb, Zn), the most basic information necessary for the development of Pb-free micro-soldering alloys, were studied using the CALPHAD method. Thermodynamic analyses for describing the Gibbs energies of the constituent phases were made by optimizing the obtained data on the experimental phase diagrams, and such data in the literature, including data on thermochemical properties. The present results combined with the thermodynamic database which was recently developed by our group [I. Ohnuma et al., J. Electron. Mater. 28, 1164 (1999)] provide various information on phase equilibria such as liquidus and solidus surfaces, isothermal and vertical section diagrams, mole fractions of the phase constitutions, etc., and thermodynamic properties such as activity, heat of mixing, surface energy, viscosity, etc., in multi-component soldering alloy systems including the elements of Pb, Pi, Sn, Sb, Cu, Ag, Zn, and In. Typical examples for the phase diagrams and thermodynamic properties of Sn-In-X ternary systems are shown. The application of the database to the alloy design for Pb-free solders is also presented

Control of neural crest induction by MarvelD3-mediated attenuation of JNK signalling

Tight junctions are required for the formation of tissue barriers and function as suppressors of signalling mechanisms that control gene expression and cell behaviour; however, little is known about the physiological and developmental importance of such signalling functions. Here, we demonstrate that depletion of MarvelD3, a transmembrane protein of tight junctions, disrupts neural crest formation and, consequently, development of neural crest-derived tissues during Xenopus embryogenesis. Using embryos and explant cultures combined with a small molecule inhibitor or mutant mRNAs, we show that MarvelD3 is required to attenuate JNK signalling during neural crest induction and that inhibition of JNK pathway activation is sufficient to rescue the phenotype induced by MarvelD3 depletion. Direct JNK stimulation disrupts neural crest development, supporting the importance of negative regulation of JNK. Our data identify the junctional protein MarvelD3 as an essential regulator of early vertebrate development and neural crest induction and, thereby, link tight junctions to the control and timing of JNK signalling during early development

Phase equilibria in the Cu-Fe-Mo and Cu-Fe-Nb systems

Phase equilibria in the Cu-Fe portion of the Cu-Fe-Mo and the Cu-Fe-Nb systems, In the temperature ranges 1073 to 1573 K and 1373 to 1573 K, respectively, were determined by metallography and scanning electron microscopy-energy dispersive x-ray methods. Based on the present experimental data combined with the previous assessments of the component binary systems, thermodynamic calculations of phase equilibria were carried out adopting the subregular solution model to describe the Gibbs energies of the liquid, bcc, and fee phases. The evaluated thermodynamic parameters lead to a better fit between calculations and experimental data in both the Cu-Fe-Mo and Cu-Fe-Nb systems