Differentiation of Human Embryonic Stem Cells into Cells with Corneal Keratocyte Phenotype

Corneal transparency depends on a unique extracellular matrix secreted by stromal keratocytes, mesenchymal cells of neural crest lineage. Derivation of keratocytes from human embryonic stem (hES) cells could elucidate the keratocyte developmental pathway and open a potential for cell-based therapy for corneal blindness. This study seeks to identify conditions inducing differentiation of pluripotent hES cells to the keratocyte lineage. Neural differentiation of hES cell line WA01(H1) was induced by co-culture with mouse PA6 fibroblasts. After 6 days of co-culture, hES cells expressing cell-surface NGFR protein (CD271, p75NTR) were isolated by immunoaffinity adsorption, and cultured as a monolayer for one week. Keratocyte phenotype was induced by substratum-independent pellet culture in serum-free medium containing ascorbate. Gene expression, examined by quantitative RT-PCR, found hES cells co-cultured with PA6 cells for 6 days to upregulate expression of neural crest genes including NGFR, SNAI1, NTRK3, SOX9, and MSX1. Isolated NGFR-expressing cells were free of PA6 feeder cells. After expansion as a monolayer, mRNAs typifying adult stromal stem cells were detected, including BMI1, KIT, NES, NOTCH1, and SIX2. When these cells were cultured as substratum-free pellets keratocyte markers AQP1, B3GNT7, PTDGS, and ALDH3A1 were upregulated. mRNA for keratocan (KERA), a cornea-specific proteoglycan, was upregulated more than 10,000 fold. Culture medium from pellets contained high molecular weight keratocan modified with keratan sulfate, a unique molecular component of corneal stroma. These results show hES cells can be induced to differentiate into keratocytes in vitro. Pluripotent stem cells, therefore, may provide a renewable source of material for development of treatment of corneal stromal opacities. © 2013 Chan et al

A desalination guide for South African municipal engineers

Municipalities need to develop Water Services Development Plans (WSDPs) as a first requirement in their budgetary process, and need to be made aware of the options that are available to provide more than basic services. While 25./personEd has been set as the minimum basic water supply and while many consumers receive far in excess of this amount, there are areas of the country where water of acceptable quality is not available for household use. However, in many areas adequate quantities of saline water may be, or are, readily available. This is especially the case for coastal cities and towns.The cost of treating water is only part of the total cost of making drinking water available to the consumer. This together with the fact that membrane technology is becoming more affordable and that energy can be recovered, makes the desalination of water viable for domestic purposes.A desk study, funded by the Department Water Affairs and Forestry and managed by the Water Research Commission, was undertaken to identify treatment options for desalinating seawater from both the Indian and Atlantic oceans or brackish water from boreholes. The specific objectives of the project were to compile a Guide on the technologies that can be implemented in South Africa to treat saline water to drinking water standards, to identify the pretreatment that is necessary, and to present guidelines on operational, maintenance, management and environmental aspects relevant to the selection and use of these technologies. An important aspect was also to quantify the capital and operating costs for planning purposes of the different components needed to successfully bring the water to the accepted standards for potable and domestic use.Of particular importance for the South African application was to identify the level of skills required for daily operation of the desalination plants, the level of skills required to provide technical back-up and advice, and to identify and advise on the competencies, training needs and capacity building required at operator and management levels. Lastly, the relevant local environmental legislations governing desalination were also identified

In-Situ Li-Ion Pouch Cell Diagnostics Utilising Plasmonic Based Optical Fibre Sensors

As the drive to improve the cost, performance characteristics and safety of lithium-ion batteries increases with adoption, one area where significant value could be added is that of battery diagnostics. This paper documents an investigation into the use of plasmonic-based optical fibre sensors, inserted internally into 1.4 Ah lithium-ion pouch cells, as a real time and in-situ diagnostic technique. The successful implementation of the fibres inside pouch cells is detailed and promising correlation with battery state is reported, while having negligible impact on cell performance in terms of capacity and columbic efficiency. The testing carried out includes standard cycling and galvanostatic intermittent titration technique (GITT) tests, and the use of a reference electrode to correlate with the anode and cathode readings separately. Further observations are made around the sensor and analyte interaction mechanisms, robustness of sensors and suggested further developments. These finding show that a plasmonic-based optical fibre sensor may have potential as an opto-electrochemical diagnostic technique for lithium-ion batteries, offering an unprecedented view into internal cell phenomena

Solar wind speed theory and the nonextensivity of solar corona

The solar corona is a complex system, with nonisothermal plasma and being in the self-gravitating field of the Sun. So the corona plasma is not only a nonequilibrium system but also a nonextensive one. We estimate the parameter of describing the degree of nonextensivity of the corona plasma and study the generalization of the solar wind speed theory in the framework of nonextensive statistical mechanics. It is found that, when use Chapman's corona model (1957) as the radial distribution of the temperature in the corona, the nonextensivity reduces the gas pressure outward and thus leads a significant deceleration effect on the radial speed of the solar wind.Comment: 12 pages,1 figure, 1 table, 21 references; UN/ESA/NASA Workshop on Basic Space Science and the International Heliophysical Year 2007, National Astronomical Observatory of Japan, 18-22 June, 2007, Tokyo, Japa

Controlled interfacial assembly of 2D curved colloidal crystals and jammed shells

Assembly of colloidal particles on fluid interfaces is a promising technique for synthesizing two-dimensional micro-crystalline materials useful in fields as diverse as biomedicine1, materials science2, mineral flotation3 and food processing4. Current approaches rely on bulk emulsification methods, require further chemical and thermal treatments, and are restrictive with respect to the materials employed5-9. The development of methods that exploit the great potential of interfacial assembly for producing tailored materials have been hampered by the lack of understanding of the assembly process. Here we report a microfluidic method that allows direct visualization and understanding of the dynamics of colloidal crystal growth on curved interfaces. The crystals are periodically ejected to form stable jammed shells, which we refer to as colloidal armour. We propose that the energetic barriers to interfacial crystal growth and organization can be overcome by targeted delivery of colloidal particles through hydrodynamic flows. Our method allows an unprecedented degree of control over armour composition, size and stability.Comment: 18 pages, 5 figure

Belowground DNA-based techniques: untangling the network of plant root interactions

Contains fulltext : 91591.pdf (publisher's version ) (Closed access)7 p

On single and double soft behaviors in NLSM

In this paper, we study the single and double soft behaviors of tree level off-shell currents and on-shell amplitudes in nonlinear sigma model(NLSM). We first propose and prove the leading soft behavior of the tree level currents with a single soft particle. In the on-shell limit, this single soft emission becomes the Adler's zero. Then we establish the leading and sub-leading soft behaviors of tree level currents with two adjacent soft particles. With a careful analysis of the on-shell limit, we obtain the double soft behaviors of on-shell amplitudes where the two soft particles are adjacent to each other. By applying Kleiss-Kuijf (KK) relation, we further obtain the leading and sub-leading behaviors of amplitudes with two nonadjacent soft particles.Comment: 41 pages, 6 tables, 9 figures, minor revised, more content about nonadjacent double soft limit, update the reference

Validation of reference genes for quantitative RT-PCR studies in porcine oocytes and preimplantation embryos

<p>Abstract</p> <p>Background</p> <p>In the developing embryo, total RNA abundance fluctuates caused by functional RNA degradation and zygotic genome activation. These variations in the transcriptome in early development complicate the choice of good reference genes for gene expression studies by quantitative real time polymerase chain reaction.</p> <p>Results</p> <p>In order to identify stably expressed genes for normalisation of quantitative data, within early stages of development, transcription levels were examined of 7 frequently used reference genes (<it>B2M, BACT, GAPDH, H2A, PGK1, SI8</it>, and <it>UBC</it>) at different stages of early porcine embryonic development (germinal vesicle, metaphase-2, 2-cell, 4-cell, early blastocyst, expanded blastocyst). Analysis of transcription profiling by geNorm software revealed that <it>GAPDH, PGK1, S18</it>, and <it>UBC </it>showed high stability in early porcine embryonic development, while transcription levels of <it>B2M, BACT</it>, and <it>H2A </it>were highly regulated.</p> <p>Conclusion</p> <p>Good reference genes that reflect total RNA content were identified in early embryonic development from oocyte to blastocyst. A selection of either <it>GAPDH </it>or <it>PGK1</it>, together with ribosomal protein <it>S18 </it>(<it>S18</it>), and <it>UBC </it>is proposed as reference genes, but the use of <it>B2M, BACT</it>, or <it>H2A </it>is discouraged.</p

Nitisinone Arrests but Does Not Reverse Ochronosis in Alkaptonuric Mice.

Alkaptonuria (AKU) is an ultrarare autosomal recessive disorder resulting from a deficiency of homogentisate 1,2 dioxygenase (HGD), an enzyme involved in the catabolism of phenylalanine and tyrosine. Loss of HGD function prevents metabolism of homogentisic acid (HGA), leading to increased levels of plasma HGA and urinary excretion. Excess HGA becomes deposited in collagenous tissues and subsequently undergoes polymerisation, principally in the cartilages of loaded joints, in a process known as ochronosis. This results in an early-onset, devastating osteoarthropathy for which there is currently no effective treatment. We recently described the natural history of ochronosis in a murine model of AKU, demonstrating that deposition of ochronotic pigment begins very early in life and accumulates with age. Using this model, we were able to show that lifetime treatment with nitisinone, a potential therapy for AKU, was able to completely prevent deposition of ochronotic pigment. However, although nitisinone has been shown to inhibit ochronotic deposition, whether it can also facilitate removal of existing pigment has not yet been examined. We describe here that midlife administration of nitisinone to AKU mice arrests further deposition of ochronotic pigment in the tibiofemoral joint, but does not result in the clearance of existing pigment. We also demonstrate the dose-dependent response of plasma HGA to nitisinone, highlighting its efficacy for personalised medicine, where dosage can be tailored to the individual AKU patient