Wide awake and ready to move: 20 years of non-viral therapeutic genome engineering with the Sleeping Beauty transposon system

Gene therapies will only become a widespread tool in the clinical treatment of human diseases with the advent of gene transfer vectors that integrate genetic information stably, safely, effectively, and economically. Two decades after the discovery of the Sleeping Beauty (SB) transposon, it has been transformed into a vector system that is fulfilling these requirements. SB may well overcome some of the limitations associated with viral gene transfer vectors and transient non-viral gene delivery approaches that are being used in the majority of ongoing clinical trials. The SB system has achieved a high level of stable gene transfer and sustained transgene expression in multiple primary human somatic cell types, representing crucial steps that may permit its clinical use in the near future. This article reviews the most important aspects of SB as a tool for gene therapy, including aspects of its vectorization and genomic integration. As an illustration, the clinical development of the SB system toward gene therapy of age-related macular degeneration and cancer immunotherapy is highlighted

A Fuzzy Logic Controller for Autonomous Wheeled Vehicles

Autonomous vehicles have potential applications in many fields, such as replacing humans in hazardous environments, conducting military missions, and performing routine tasks for industry. Driving ground vehicles is an area where human performance has proven to be reliable. Drivers typically respond quickly to sudden changes in their environment. While other control techniques may be used to control a vehicle, fuzzy logic has certain advantages in this area; one of them is its ability to incorporate human knowledge and experience, via language, into relationships among the given quantities. Fuzzy logic controllers for autonomous vehicles have been successfully applied to address various (and sometimes simultaneous) navigational issues

The Pulsed Neutron Beam EDM Experiment

We report on the Beam EDM experiment, which aims to employ a pulsed cold neutron beam to search for an electric dipole moment instead of the established use of storable ultracold neutrons. We present a brief overview of the basic measurement concept and the current status of our proof-of-principle Ramsey apparatus

History-dependent relaxation and the energy scale of correlation in the Electron-Glass

We present an experimental study of the energy-relaxation in Anderson-insulating indium-oxide films excited far from equilibrium. In particular, we focus on the effects of history on the relaxation of the excess conductance dG. The natural relaxation law of dG is logarithmic, namely dG=-log(t). This may be observed over more than five decades following, for example, cool-quenching the sample from high temperatures. On the other hand, when the system is excited from a state S_{o} in which it has not fully reached equilibrium to a state S_{n}, the ensuing relaxation law is logarithmic only over time t shorter than the time t_{w} it spent in S_{o}. For times t>t_{w} dG(t) show systematic deviation from the logarithmic dependence. It was previously shown that when the energy imparted to the system in the excitation process is small, this leads to dG=P(t/t_{w}) (simple-aging). Here we test the conjecture that `simple-aging' is related to a symmetry in the relaxation dynamics in S_{o} and S_{n}. This is done by using a new experimental procedure that is more sensitive to deviations in the relaxation dynamics. It is shown that simple-aging may still be obeyed (albeit with a modified P(t/t_{w})) even when the symmetry of relaxation in S_{o} and S_{n} is perturbed by a certain degree. The implications of these findings to the question of aging, and the energy scale associated with correlations are discussed

The Glass Transition Temperature of Water: A Simulation Study

We report a computer simulation study of the glass transition for water. To mimic the difference between standard and hyperquenched glass, we generate glassy configurations with different cooling rates and calculate the $T$ dependence of the specific heat on heating. The absence of crystallization phenomena allows us, for properly annealed samples, to detect in the specific heat the simultaneous presence of a weak pre-peak (``shadow transition''), and an intense glass transition peak at higher temperature. We discuss the implications for the currently debated value of the glass transition temperature of water. We also compare our simulation results with the Tool-Narayanaswamy-Moynihan phenomenological model.Comment: submitted to Phys. Re

The JAK inhibitor tofacitinib suppresses synovial JAK1-STAT signalling in rheumatoid arthritis.

ObjectiveTofacitinib is an oral Janus kinase (JAK) inhibitor for the treatment of rheumatoid arthritis (RA). The pathways affected by tofacitinib and the effects on gene expression in situ are unknown. Therefore, tofacitinib effects on synovial pathobiology were investigated.MethodsA randomised, double-blind, phase II serial synovial biopsy study (A3921073; NCT00976599) in patients with RA with an inadequate methotrexate response. Patients on background methotrexate received tofacitinib 10 mg twice daily or placebo for 28 days. Synovial biopsies were performed on Days -7 and 28 and analysed by immunoassay or quantitative PCR. Clinical response was determined by disease activity score and European League Against Rheumatism (EULAR) response on Day 28 in A3921073, and at Month 3 in a long-term extension study (A3921024; NCT00413699).ResultsTofacitinib exposure led to EULAR moderate to good responses (11/14 patients), while placebo was ineffective (1/14 patients) on Day 28. Tofacitinib treatment significantly reduced synovial mRNA expression of matrix metalloproteinase (MMP)-1 and MMP-3 (p<0.05) and chemokines CCL2, CXCL10 and CXCL13 (p<0.05). No overall changes were observed in synovial inflammation score or the presence of T cells, B cells or macrophages. Changes in synovial phosphorylation of signal transducer and activator of transcription 1 (STAT1) and STAT3 strongly correlated with 4-month clinical responses (p<0.002). Tofacitinib significantly decreased plasma CXCL10 (p<0.005) at Day 28 compared with placebo.ConclusionsTofacitinib reduces metalloproteinase and interferon-regulated gene expression in rheumatoid synovium, and clinical improvement correlates with reductions in STAT1 and STAT3 phosphorylation. JAK1-mediated interferon and interleukin-6 signalling likely play a key role in the synovial response.Trial registration numberNCT00976599

Biomaterials for corneal bioengineering

© 2018 IOP Publishing Ltd. Corneal transplantation is an important surgical treatment for many common corneal diseases. However, a worldwide shortage of tissue from suitable corneal donors has meant that many people are not able to receive sight-restoring operations. In addition, rejection is a major cause of corneal transplant failure. Bioengineering corneal tissue has recently gained widespread attention. In order to facilitate corneal regeneration, a range of materials is currently being investigated. The ideal substrate requires sufficient tectonic durability, biocompatibility with cultured cellular elements, transparency, and perhaps biodegradability and clinical compliance. This review considers the anatomy and function of the native cornea as a precursor to evaluating a variety of biomaterials for corneal regeneration including key characteristics for optimal material form and function. The integration of appropriate cells with the most appropriate biomaterials is also discussed. Taken together, the information provided offers insight into the requirements for fabricating synthetic and semisynthetic corneas for in vitro modeling of tissue development and disease, pharmaceutical screening, and in vivo application for regenerative medicine

Exopolysaccharides extracted from Parachlorella kessleri inhibit colon carcinoma growth in mice via stimulation of host antitumor immune responses

Citation: Ishiguro, S., Uppalapati, D., Goldsmith, Z., Robertson, D., Hodge, J., Holt, H., . . . Tamura, M. (2017). Exopolysaccharides extracted from Parachlorella kessleri inhibit colon carcinoma growth in mice via stimulation of host antitumor immune responses. Plos One, 12(4), 21. https://doi.org/10.1371/journal.pone.0175064The newly purified extracellular polysaccharides (exopolysaccharides) from Parachlorella kessleri (PCEPS) were evaluated on their antitumor and immunomodulatory effects in cell culture and mouse colon carcinoma peritoneal dissemination model. In two-dimensional cell culture, the PCEPS treatment inhibited cell growth of both murine and human colon carcinoma cells in a dose- and time-dependent manner. In contrast, the growth of mouse splenocytes (SPLs) and bone marrow cells (BMCs) were stimulated by the treatment with PCEPS. The treatment with PCEPS also increased specific subpopulations of the cells in BMCs: antigen presenting cells (CD19(+) B cells, 33D1(+) dendritic cells and CD68(+) macrophage) and CD8(+) cytotoxic T cells. In three-dimensional spheroid culture, spheroid growth of CT26 cells co-cultured with HL-60 human neutrophilic promyeloblasts and Jurkat cells (human lymphoblasts), but not THP1 human monocyte/macrophage was significantly attenuated by PCEPS treatment. In a mouse CT26 colon carcinoma peritoneal dissemination model, intraperitoneal injection of PCEPS (10 mg/kg, twice per week) significantly attenuated the growth of CT26 colon carcinoma in syngeneic mice. The present study suggests that PCEPS inhibits colon carcinoma growth via direct cell growth inhibition and a stimulation of the host antitumor immune responses. Taken together, the current study suggests that exopolysaccharides derived from Parachlorella kessleri contain significant bioactive materials that inhibit colon carcinoma growth

A Survey of Local Group Galaxies Currently Forming Stars. I. UBVRI Photometry of Stars in M31 and M33

We present UBVRI photometry obtained from Mosaic images of M31 and M33 using the KPNO 4-m telescope. The survey covers 2.2 sq degrees of M31, and 0.8 sq degrees of M33, chosen so as to include all of the regions currently active in forming massive stars. The catalog contains 371,781 and 146,622 stars in M31 and M33, respectively, where every star has a counterpart (at least) in B, V, and R. We compare our photometry to previous studies. We provide cross references to the stars confirmed as members by spectroscopy, and compare the location of these to the complete set in color-magnitude diagrams. While follow-up spectroscopy is needed for many projects, we demonstrate the success of our photometry in being able to distinguish M31/M33 members from foreground Galactic stars. We also present the results of newly obtained spectroscopy, which identifies 34 newly confirmed members, including B-A supergiants, the earliest O star known in M31, and two new Luminous Blue Variable candidates whose spectra are similar to that of P Cygni.Comment: Accepted by the Astronomical Journal. A version with higher resolution figures can be found at: http://www.lowell.edu/users/massey/M3133.pdf.g