Oral mucosal epithelial cells grown on porous silicon membrane for transfer to the rat eye

Dysfunction of limbal stem cells or their niche can result in painful, potentially sight-threatening ocular surface disease. We examined the utility of surface-modified porous-silicon (pSi) membranes as a scaffold for the transfer of oral mucosal cells to the eye. Male-origin rat oral mucosal epithelial cells were grown on pSi coated with collagen-IV and vitronectin, and characterised by immunocytochemistry. Scaffolds bearing cells were implanted into normal female rats, close to the limbus, for 8 weeks. Histology, immunohistochemistry and a multiplex nested PCR for sry were performed to detect transplanted cells. Oral mucosal epithelial cells expanded on pSi scaffolds expressed the corneal epithelial cell marker CK3/12. A large percentage of cells were p63⁺, indicative of proliferative potential, and a small proportion expressed ABCG2⁺, a putative stem cell marker. Cell-bearing scaffolds transferred to the eyes of live rats, were well tolerated, as assessed by endpoint histology. Immunohistochemistry for pan-cytokeratins demonstrated that transplanted epithelial cells were retained on the pSi membranes at 8 weeks post-implant, but were not detectable on the central cornea using PCR for sry. The pSi scaffolds supported and retained transplanted rat oral mucosal epithelial cells in vitro and in vivo and recapitulate some aspects of an artificial stem cell niche.Yazad D. Irani, Sonja Klebe, Steven J.P. McInnes, Marek Jasieniak, Nicolas H. Voelcker, Keryn A. William

Toll-Like Receptor Expression in Human Keratinocytes: Nuclear Factor κB Controlled Gene Activation by Staphylococcus aureus is Toll-Like Receptor 2 But Not Toll-Like Receptor 4 or Platelet Activating Factor Receptor Dependent

Cultured primary human keratinocytes were screened for their expression of various members of the toll-like receptor (TLR) family. Keratinocytes were found to constitutively express TLR1, TLR2, TLR3, TLR5, and TLR9 but not TLR4, TLR6, TLR7, TLR8, or TLR10 as shown by polymerase chain reaction analysis. The expression of the crucial receptor for signaling of staphylococcal compounds TLR2 was also confirmed by immunohistochemistry, in contrast to TLR4, which showed a negative staining pattern. Next, we analyzed the activation of the proinflammatory nuclear transcription factor κB by Staphylococcus aureus strain 8325-4. Using nuclear extract gel shifts, RelA staining, and luciferase reporter transfection plasmids we found a clear induction of nuclear factor κB translocation by the bacteria. This translocation induced the transcription of nuclear factor κB controlled genes such as inducible nitric oxide synthetase, COX2, and interleukin-8. Transcription of these genes was followed by production of increased amounts of interleukin-8 protein and NO. Inhibition experiments using monoclonal antibodies and the specific platelet activating factor receptor inhibitor CV3988 showed that nuclear factor κB activation by S. aureus was TLR2 but not TLR4 or platelet activating factor receptor dependent. In line, the purified staphylococcal cell wall components lipoteichoic acid and peptidoglycan, known to signal through TLR2, also showed nuclear factor κB translocation in human keratinocytes, indicating a crucial role of the staphylococcal cell wall in the innate immune stimulation of human keratinocytes. These results help to explain the complex activation of human keratinocytes by S. aureus and its cell wall components in various inflammatory disorders of the skin

Non-viral gene therapy that targets motor neurons in vivo

A major challenge in neurological gene therapy is safe delivery of transgenes to sufficient cell numbers from the circulation or periphery. This is particularly difficult for diseases involving spinal cord motor neurons such as amyotrophic lateral sclerosis (ALS). We have examined the feasibility of non-viral gene delivery to spinal motor neurons from intraperitoneal injections of plasmids carried by "immunogene" nanoparticles targeted for axonal retrograde transport using antibodies. PEGylated polyethylenimine (PEI-PEG12) as DNA carrier was conjugated to an antibody (MLR2) to the neurotrophin receptor p75 (p75NTR). We used a plasmid (pVIVO2) designed for in vivo gene delivery that produces minimal immune responses, has improved nuclear entry into post mitotic cells and also expresses green fluorescent protein (GFP). MLR2-PEI-PEG12 carried pVIVO2 and was specific for mouse motor neurons in mixed cultures containing astrocytes. While only 8% of motor neurons expressed GFP 72 h post transfection in vitro, when the immunogene was given intraperitonealy to neonatal C57BL/6J mice, GFP specific motor neuron expression was observed in 25.4% of lumbar, 18.3% of thoracic and 17.0% of cervical motor neurons, 72 h post transfection. PEI-PEG12 carrying pVIVO2 by itself did not transfect motor neurons in vivo, demonstrating the need for specificity via the p75NTR antibody MLR2. This is the first time that specific transfection of spinal motor neurons has been achieved from peripheral delivery of plasmid DNA as part of a non-viral gene delivery agent. These results stress the specificity and feasibility of immunogene delivery targeted for p75NTR expressing motor neurons, but suggests that further improvements are required to increase the transfection efficiency of motor neurons in vivo.Mary-Louise Rogers, Kevin S. Smith, Dusan Matusica, Matthew Fenech, Lee Hoffman, Robert A. Rush and Nicolas H. Voelcke

A Radial Velocity Survey of the Cygnus OB2 Association

We conducted a radial velocity survey of the Cygnus OB2 Association over a 6 year (1999 - 2005) time interval to search for massive close binaries. During this time we obtained 1139 spectra on 146 OB stars to measure mean systemic radial velocities and radial velocity variations. We spectroscopically identify 73 new OB stars for the first time, the majority of which are likely to be Association members. Spectroscopic evidence is also presented for a B3Iae classification and temperature class variation (B3 - B8) on the order of 1 year for Cygnus OB2 No. 12. Calculations of the intial mass function with the current spectroscopic sample yield Gamma = -2.2 +/- 0.1. Of the 120 stars with the most reliable data, 36 are probable and 9 are possible single-lined spectroscopic binaries. We also identify 3 new and 8 candidate double-lined spectroscopic binaries. These data imply a lower limit on the massive binary fraction of 30% - 42%. The calculated velocity dispersion for Cygnus OB2 is 2.44 +/- km/s, which is typical of open clusters. No runaway OB stars were found.Comment: 56 pages, 23 figures, 5 tables, accepted for publication in the Astrophysical Journa

Designing superhydrophobic surfaces using fluorosilsesquioxane-urethane hybrid and porous silicon gradients

Bellingham, Washingto

Nitric oxide releasing plasma polymer coating with bacteriostatic properties and no cytotoxic side effects

Published on 19 March 2015We report a stable plasma polymer coating, using isopentyl nitrite as a volatile precursor, which releases nitric oxide at bacteriostatic concentrations when contacted with water, inhibiting bacterial growth without cytotoxic side effects to human mesenchymal stem/stromal cells.Thomas D. Michl, Bryan R. Coad, Michael Doran, Michael Osiecki, Morteza Hasanzadeh Kafshgari, Nicolas H. Voelcker, Amanda Hüsler, Krasimir Vasilev and Hans J. Griesse

Physical activity to improve cognition in older adults: can physical activity programs enriched with cognitive challenges enhance the effects? A systematic review and meta-analysis

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Thiol-capped gold nanoparticles swell-encapsulated into polyurethane as powerful antibacterial surfaces under dark and light conditions

A simple procedure to develop antibacterial surfaces using thiol-capped gold nanoparticles (AuNPs) is shown, which effectively kill bacteria under dark and light conditions. The effect of AuNP size and concentration on photo-activated antibacterial surfaces is reported and we show significant size effects, as well as bactericidal activity with crystal violet (CV) coated polyurethane. These materials have been proven to be powerful antibacterial surfaces against both Gram-positive and Gram-negative bacteria. AuNPs of 2, 3 or 5 nm diameter were swell-encapsulated into PU before a coating of CV was applied (known as PU-AuNPs-CV). The antibacterial activity of PU-AuNPs-CV samples was tested against Staphylococcus aureus and Escherichia coli as representative Gram-positive and Gram-negative bacteria under dark and light conditions. All light conditions in this study simulated a typical white-light hospital environment. This work demonstrates that the antibacterial activity of PU-AuNPs-CV samples and the synergistic enhancement of photoactivity of triarylmethane type dyes is highly dependent on nanoparticle size and concentration. The most powerful PU-AuNPs-CV antibacterial surfaces were achieved using 1.0 mg mL−1 swell encapsulation concentrations of 2 nm AuNPs. After two hours, Gram-positive and Gram-negative bacteria were reduced to below the detection limit (>4 log) under dark and light conditions

Will the Playstation generation become better endoscopic surgeons?

A frequently heard comment is that the current "Playstation generation" will have superior baseline psychomotor skills. However, research has provided inconsistent results on this matter. The purpose of this study was to investigate whether the "Playstation generation" shows superior baseline psychomotor skills for endoscopic surgery on a virtual reality simulator. The 46 study participants were interns (mean age 24 years) of the department of surgery and schoolchildren (mean age 12.5 years) of the first year of a secondary school. Participants were divided into four groups: 10 interns with videogame experience and 10 without, 13 schoolchildren with videogame experience and 13 without. They performed four tasks twice on a virtual reality simulator for basic endoscopic skills. The one-way analysis of variance (ANOVA) with post hoc test Tukey-Bonferroni and the independent Student's t test were used to determine differences in mean scores. Interns with videogame experience scored significantly higher on total score (93 vs. 74.5; p=0.014) compared with interns without this experience. There was a nonsignificant difference in mean total scores between the group of schoolchildren with and those without videogame experience (61.69 vs. 55.46; p=0.411). The same accounts for interns with regard to mean scores on efficiency (50.7 vs. 38.9; p=0.011) and speed (18.8 vs. 14.3; p=0.023). In the group of schoolchildren, there was no statistical difference for efficiency (32.69 vs. 27.31; p=0.218) or speed (13.92 vs. 13.15; p=0.54). The scores concerning precision parameters did not differ for interns (23.5 vs. 21.3; p=0.79) or for schoolchildren (mean 15.08 vs. 15; p=0.979). Our study results did not predict an advantage of videogame experience in children with regard to superior psychomotor skills for endoscopic surgery. However, at adult age, a difference in favor of gaming is present. The next generation of surgeons might benefit from videogame experience during their childhoo