A novel splice variant of the DNA-PKcs gene is associated with clinical and cellular radiosensivity in a patient with xeroderma pigmentosum

Background: Radiotherapy-induced DNA double-strand breaks (DSBs) are critical cytotoxic lesions. Inherited defects in DNA DSB repair pathways lead to hypersensitivity to ionising radiation, immunodeficiency and increased cancer incidence. A patient with xeroderma pigmentosum complementation group C, with a scalp angiosarcoma, exhibited dramatic clinical radiosensitivity following radiotherapy, resulting in death. A fibroblast cell line from non-affected skin (XP14BRneo17) was hypersensitive to ionising radiation and defective in DNA DSB repair. Aim: To determine the genetic defect causing cellular radiation hypersensitivity in XP14BRneo17 cells. Methods: Functional genetic complementation whereby copies of human chromosomes containing genes involved in DNA DSB repair (chromosomes 2, 5, 8 10, 13 and 22) were individually transferred to XP14BRneo17 cells in an attempt to correct the radiation hypersensitivity. Clonogenic survival assays and g-H2AX immunofluorescence were conducted to measure radiation sensitivity and repair of DNA DSBs. DNA sequencing of defective DNA repair genes was performed. Results: Transfer of chromosome 8 (location of DNAPKcs gene) and transfection of a mammalian expression construct containing the DNA-PKcs cDNA restored normal ionising radiation sensitivity and repair of DNA DSBs in XP14BRneo17 cells. DNA sequencing of the DNA-PKcs coding region revealed a 249-bp deletion (between base pairs 3656 and 3904) encompassing exon 31 of the gene. Conclusion: We provide evidence of a novel splice variant of the DNA-PKcs gene associated with radiosensitivity in a patient with xeroderma pigmentosum and report the first double mutant in distinct DNA repair pathways being consistent with viability

Developing Optimized Trajectories Derived from Mission and Thermo-Structural Constraints

In conjunction with NASA and the Department of Defense, the Johns Hopkins University Applied Physics Laboratory (JHU/APL) has been investigating analytical techniques to address many of the fundamental issues associated with solar exploration spacecraft and high-speed atmospheric vehicle systems. These issues include: thermo-structural response including the effects of thermal management via the use of surface optical properties for high-temperature composite structures; aerodynamics with the effects of non-equilibrium chemistry and gas radiation; and aero-thermodynamics with the effects of material ablation for a wide range of thermal protection system (TPS) materials. The need exists to integrate these discrete tools into a common framework that enables the investigation of interdisciplinary interactions (including analysis tool, applied load, and environment uncertainties) to provide high fidelity solutions. In addition to developing robust tools for the coupling of aerodynamically induced thermal and mechanical loads, JHU/APL has been studying the optimal design of high-speed vehicles as a function of their trajectory. Under traditional design methodology the optimization of system level mission parameters such as range and time of flight is performed independently of the optimization for thermal and mechanical constraints such as stress and temperature. A truly optimal trajectory should optimize over the entire range of mission and thermo-mechanical constraints. Under this research, a framework for the robust analysis of high-speed spacecraft and atmospheric vehicle systems has been developed. It has been built around a generic, loosely coupled framework such that a variety of readily available analysis tools can be used. The methodology immediately addresses many of the current analysis inadequacies and allows for future extension in order to handle more complex problems

Synthetic poly(ester amine) and poly(amido amine) nanoparticles for efficient DNA and siRNA delivery to human endothelial cells

Biodegradable poly(ester amine) (PEA)-based and poly(amido amine) (PAA)-based nanoparticles were developed for efficient in vitro siRNA delivery to human umbilical vein endothelial cells (HUVECs). They were screened, characterized, and compared with traditionally studied DNA-containing particles. Several of the polymeric nanoparticles tested were found to be effective for delivering functional siRNA to green fluorescent protein (GFP) + HUVECs, achieving 60%–75% GFP knockdown while maintaining high viability. While PEAs have been used previously to form polyplexes or nanoparticles for DNA delivery, highly effective siRNA delivery in hard-to-transfect human cell types has not been previously reported. PEAs and linear nondendrimeric PAAs were also found to be effective for DNA delivery to HUVECs using GFP-encoding plasmid DNA (up to 50%–60% transfection efficiency). PEAs and PAAs can be separated into groups that form polymeric nanoparticles effective for siRNA delivery, for DNA delivery, or for both

Celiac disease and risk of myasthenia gravis – nationwide population-based study

Background Case reports suggest there may be an association between celiac disease (CD) and myasthenia gravis (MG). Methods We identified 29,086 individuals with CD in Sweden from 1969 to 2008. We compared these individuals with 144,480 matched controls. Hazard ratios (HRs) for future MG (identified through ICD codes) were estimated using Cox regression. Results During 326,376 person-years of follow-up in CD patients, there were 7 MG cases (21/million person-years) compared to 22 MG cases in controls during 1,642,273 years of follow-up (14/million person-years) corresponding to a HR of 1.48 (95% CI = 0.64–3.41). HRs did not differ when stratifying for age, sex or calendar period. HRs were highest in the first year after follow-up, though insignificant. Individuals with CD were at no increased risk of MG more than 5 years after CD diagnosis (HR = 0.70; 95% CI = 0.16–3.09). Conclusion This study found no increased risk of MG in patients with CD

Transforming growth factor-beta promotes rhinovirus replication in bronchial epithelial cells by suppressing the innate immune response

Rhinovirus (RV) infection is a major cause of asthma exacerbations which may be due to a deficient innate immune response in the bronchial epithelium. We hypothesized that the pleiotropic cytokine, TGF-?, influences interferon (IFN) production by primary bronchial epithelial cells (PBECs) following RV infection. Exogenous TGF-?(2) increased RV replication and decreased IFN protein secretion in response to RV or double-stranded RNA (dsRNA). Conversely, neutralizing TGF-? antibodies decreased RV replication and increased IFN expression in response to RV or dsRNA. Endogenous TGF-?(2) levels were higher in conditioned media of PBECs from asthmatic donors and the suppressive effect of anti-TGF-? on RV replication was significantly greater in these cells. Basal SMAD-2 activation was reduced when asthmatic PBECs were treated with anti-TGF-? and this was accompanied by suppression of SOCS-1 and SOCS-3 expression. Our results suggest that endogenous TGF-? contributes to a suppressed IFN response to RV infection possibly via SOCS-1 and SOCS-3

Lithium in strong magnetic fields

The electronic structure of the lithium atom in a strong magnetic field 0 <= gamma <= 10 is investigated. Our computational approach is a full configuration interaction method based on a set of anisotropic Gaussian orbitals that is nonlinearly optimized for each field strength. Accurate results for the total energies and one-electron ionization energies for the ground and several excited states for each of the symmetries ^20^+, ^2(-1)^+, ^4(-1)^+, ^4(-1)^-, ^2(-2)^+, ^4(-2)^+, $^4(-3)^{+}$ are presented. The behaviour of these energies as a function of the field strength is discussed and classified. Transition wave lengths for linear and circular polarized transitions are presented as well.Comment: 12 pages, 13 figures, accepted for publication in Phys. Rev.

Regular solutions to higher order curvature Einstein--Yang-Mills systems in higher dimensions

We study regular, static, spherically symmetric solutions of Yang-Mills theories employing higher order invariants of the field strength coupled to gravity in $d$ dimensions. We consider models with only two such invariants characterised by integers $p$ and $q$. These models depend on one dimensionless parameter $\alpha$ leading to one-parameter families of regular solutions, obtainable by numerical solution of the corresponding boundary value problem. Much emphasis is put on an analytical understanding of the numerical results.Comment: 34 pages, 12 figure