Isolation of epithelial stem cells from dermis by a three-dimensional culture system

Skin is a representative self-renewing tissue containing stem cells. Although many attempts have been made to define and isolate skin-derived stem cells, establishment of a simple and reliable isolation procedure remains a goal to be achieved. Here, we report the isolation of cells having stem cell properties from mouse embryonic skin using a simple selection method based on an assumption that stem cells may grow in an anchorage-independent manner. We inoculated single cell suspensions prepared from mouse embryonic dermis into a temperature-sensitive gel and propagated the resulting colonies in a monolayer culture. The cells named dermis-derived epithelial progenitor-1 (DEEP) showed epithelial morphology and grew rapidly to a more than 200 population doubling level over a period of 250 days. When the cells were kept confluent, they spontaneously formed spheroids and continuously grew even in spheroids. Immunostaining revealed that all of the clones were positive for the expression of cytokeratin-8, -18, -19, and E-cadherin and negative for the expression of cytokeratin-1, -5, -6, -14, -20, vimentin, nestin, a ckit. Furthermore, they expressed epithelial stem cell markers such as p63, integrin beta1, and S100A6. On exposure to TGFbeta in culture, some of DEEP-1 cells expressed alpha-smooth muscle actin. When the cells were transplanted into various organs of adult SCID mice, a part of the inoculated cell population acquired neural, hepatic, and renal cell properties. These results indicate that the cells we isolated were of epithelial stem cell origin and that our new approach is useful for isolation of multipotent stem cells from skin tissues.

Deployment on GPUs of an application in computational atomic physics

This paper describes the deployment on GPUs of PROP, a program of the 2DRMP suite which models electron collisions with H-like atoms and ions. Because performance on GPUs is better in single precision than in double precision, the numerical stability of the PROP program in single precision has been studied. The numerical quality of PROP results computed in single precision and their impact on the next program of the 2DRMP suite has been analyzed. Successive versions of the PROP program on GPUs have been developed in order to improve its performance. Particular attention has been paid to the optimization of data transfers and of linear algebra operations. Performance obtained on several architectures (including NVIDIA Fermi) are presented.

The role of SRC-CAS interactions in cellular transformation:ectopic expression of the carboxy terminus of CAS inhibits SRC-CAS interaction but has no effect on cellular transformation

Several lines of evidence indicate that the adapter molecule p130CAS (crk-associated substrate (CAS)) is required for src-mediated cellular transformation. CAS has been shown to be heavily tyrosine-phosphorylated in src-transformed cells, and genetic variants of src that are deficient in CAS binding are also unable to mediate cellular transformation. In this report, we investigated whether CAS phosphorylation and/or its association with src are required elements of the transformation process. Expression of the carboxy-terminal src binding domain of CAS in Rat 1 fibroblasts expressing a temperature-sensitive allele of v-src inhibited the formation of src-CAS complexes and also inhibited tyrosine phosphorylation of CAS. However, expression of this protein had no effect on morphological transformation, src-mediated actin rearrangements, or anchorage-independent growth of these cells when grown at the src-permissive temperature. Thus, the ability of activated src to mediate cellular transformation is either largely independent of endogenous CAS phosphorylation and/or its association with CAS or, alternatively, the carboxy-terminus of CAS may substitute for endogenous CAS in the process of src-mediated transformation.

Measuring specific parameters in pretensioned concrete members using a single testing technique

Pretensioned concrete members are designed and manufactured by using at least two materials: concrete and prestressing reinforcement. Also, two main stages must be considered: prestress transfer and member loading. Hence, the behavior of these members depends strongly on the reinforcement-to-concrete bond performance and prestress losses. In this paper, a testing technique to measure the specific parameters related with the involved phenomena is presented. The testing technique is based on the analysis of series of specimens varying in embedment length to simulate several cross sections at only one end of a pretensioned concrete member. Each specimen is characterized by means of the sequential release of the prestress transfer (detensioning) and the pull-out (loading) operation. The test provides data on prestressing force, transmission length (initial and long-term), anchorage length (without and with slip), reinforcement slips, bond stresses, longitudinal concrete strains, concrete modulus of elasticity, and prestress losses (instantaneous and time-dependent).

The Characteristics of Wave Impacts on an Oscillating Wave Surge Converter

Wave impacts on an Oscillating Wave Surge Converter are examined using experimental and numerical methods. The mechanics of the impact event are identified experimentally with the use of images recorded with a high speed camera. It is shown that it is the device which impacts the wave rather than a breaking wave impacting the device. Numerical simulations using two different approaches are used to further understand the issue. Good agreement is shown between numerical simulations and experimental measurements at 25th scale.

Strengthening timber beams with near surface mounted carbon fiber reinforced polymer rods

Many timber structures may require strengthening due to either decay and aging or an increase of load. This paper presents an experimental study in which eleven timber beams were tested, including three unstrengthened reference beams and eight beams strengthened with NSM CFRP bars. The test parameters include the position of NSM (tensile face or the bottom of the sides), the number of CFRP bars (1 or 2), and additional anchorage of NSM CFRP bars (steel wire U anchors or CFRP U strips). The test results show that the ultimate flexural strength of the timber beams were increased by 14%∼85% with an average of 47% due to NSM CFRP bar strengthening. Their deflection corresponding to the peak load was increased by 33% in average.

Novel manually made NSM FRP (MMFRP) bars for shear strengthening of RC beams

This paper presents an experimental study evaluating the effectiveness of the Near Surface Mounted (NSM) technique with innovative manually made FRP bars (MMFRP) for shear strengthening of RC beams. RC beams designed to fail in shear were tested in three-point bending. To delay the onset of MMFRP bar debonding, a new anchorage is also developed and tested. This paper reports the results of a series of tests on simply supported rectangular RC beams, strengthened in shear with MMFRP bar either with or without the proposed anchorage. The load-deflection responses of all test beams are plotted, in addition to selected strain results. Performance and the failure modes of the test beams are presented and discussed in this paper. The proposed MMFRP bars and end anchorage enhanced the shear capacity between 25 to 48% over the control specimen. Furthermore, the adoption of the proposed end anchorage of MMFRP bars significantly enhanced the ductility of the test specimens.

A coactivator role of CARM1 in the dysregulation of β-catenin activity in colorectal cancer cell growth and gene expression

Aberrant activation of Wnt/β-catenin signaling, resulting in the expression of Wnt-regulated oncogenes, is recognized as a critical factor in the etiology of colorectal cancer. Occupancy of β-catenin at promoters of Wnt target genes drives transcription, but the mechanism of β-catenin action remains poorly understood. Here, we show that CARM1 (coactivator-associated arginine methyltransferase 1) interacts with β-catenin and positively modulates β-catenin-mediated gene expression. In colorectal cancer cells with constitutively high Wnt/β-catenin activity, depletion of CARM1 inhibits expression of endogenous Wnt/β-catenin target genes and suppresses clonal survival and anchorage-independent growth. We also identified a colorectal cancer cell line (RKO) with a low basal level of β-catenin, which is dramatically elevated by treatment with Wnt3a. Wnt3a also increased the expression of a subset of endogenous Wnt target genes, and CARM1 was required for the Wnt-induced expression of these target genes and the accompanying dimethylation of arginine 17 of histone H3. Depletion of β-catenin from RKO cells diminished the Wnt-induced occupancy of CARM1 on a Wnt target gene, indicating that CARM1 is recruited to Wnt target genes through its interaction with β-catenin and contributes to transcriptional activation by mediating events (including histone H3 methylation) that are downstream from the actions of β-catenin. Therefore, CARM1 is an important positive modulator of Wnt/β-catenin transcription and neoplastic transformation, and may thereby represent a novel target for therapeutic intervention in cancers involving aberrantly activated Wnt/β-catenin signaling.

Content and methodology of predoctoral Geriatric Dentistry teaching in Europe

Objectives: To explore the content and methodology of predoctoral Geriatric Dentistry teaching amongst European dental schools.
Methods: The study was conducted by the European College of Gerodontology (ECG) Education Committee. Αn electronic questionnaire has been developed with close and open-ended items, including information on the prevalence and institutional anchorage of Gerodontology programs, the educators, the content and the methodology of teaching. An electronic mail, including a hyperlink to the questionnaire, was sent to 216 dental schools in 39 European countries (Winter/ Spring 2016). The Deans were asked to either answer themselves, or forward the link to faculty members with knowledge on Gerodontology teaching at their respective schools. Repeated reminders or telephone calls were used for non-respondents and personal networks were exploited to identify potential contact persons.
Results: Until August 2016, 121 dental schools from 29 countries responded to the survey (response rate 56%, EU response rate: 60%). Gerodontology was included in the predoctoral curricula of 86% of the respondents and was compulsory in 68%. The course was mainly offered in senior students and was interdisciplinary in 30% of the schools, delivered mainly by dentists (79%), physicians (21%), psychologists (10%), and nurses (5%). It was conducted as an independent lecture series in 40% of the schools and a course director was assigned in 44% of the respondents. When embedded in other disciplines, these were mainly Prosthodontics (31%). The content included a large number of items, such as epidemiology of oral health, medical problems in old age, prosthodontic management, xerostomia, and caries risk assessment. Lectures were the most common teaching format (69%), followed by small group seminars (27%). The most common types of educational material used were scientific articles (48%), printed textbooks (44%), lecture notes (40%) and e-learning material (21%). Clinical training was offered by 64% of the respondents, within the dental school clinics (49%) and/or in outreach locations (40%).
Conclusion: Amongst the respondent European dental schools (66%) there is an increasing number that teach Gerodontology at a pre-doctoral level with significant variations in content and methodology. Official guidelines and the dissemination of the ECG pre-doctoral curriculum guidelines might help to increase the prevalence and improve the status of Gerodontology teaching in Europe.

The Democratic Dynamics of Government Consultations. Speaking Freely and Listening Properly

There is a growing use of consultation and e-consultation procedures by governments. This chapter seeks to examine the role of consultation as part of a new technology of government. Consultation on policy development can reinvigorate democratic engage- ment but often it can silence views through a sort of participatory disempowerment; it can loosen the democratic anchorage of the public service within the state. The chapter develops a governmentality perspective interrogating what participation, democratic engagement and free speech mean in this context, and how ideas of publicness are constructed, managed and controlled. The focus is on the nature of consultation, its relationship to ideas of free speech and speaking freely, and its potential to empower subaltern counterpublics which can formulate oppositional interpretations and urge alternative conclusions. The aim is to develop an idea of the democratic adequacy of the consultation process and draw out a sense of how democratic engagement here can be structured – for good or ill.