Rapid and efficient stable gene transfer to mesenchymal stromal cells using a modified foamy virus vector

Mesenchymal stromal cells (MSCs) hold great promise for regenerative medicine. Stable ex vivo gene transfer to MSCs could improve the outcome and scope of MSC therapy, but current vectors require multiple rounds of transduction, involve genotoxic viral promoters and/or the addition of cytotoxic cationic polymers in order to achieve efficient transduction. We describe a self-inactivating foamy virus vector (FVV), incorporating the simian macaque foamy virus envelope and using physiological promoters, which efficiently transduces murine MSCs (mMSCs) in a single-round. High and sustained expression of the transgene, whether GFP or the lysosomal enzyme, arylsulphatase A (ARSA), was achieved. Defining MSC characteristics (surface marker expression and differentiation potential), as well as long-term engraftment and distribution in the murine brain following intracerebroventricular delivery, are unaffected by FVV transduction. Similarly, greater than 95% of human MSCs (hMSCs) were stably transduced using the same vector, facilitating human application. This work describes the best stable gene transfer vector available for mMSCs and hMSCs

Assessment of DNA extracted from FTA® cards for use on the Illumina iSelect BeadChip

<p>Abstract</p> <p>Background</p> <p>As FTA^®cards provide an ideal medium for the field collection of DNA we sought to assess the quality of genomic DNA extracted from this source for use on the Illumina BovineSNP50 iSelect BeadChip which requires unbound, relatively intact (fragment sizes ≥ 2 kb), and high-quality DNA. Bovine blood and nasal swab samples collected on FTA cards were extracted using the commercially available GenSolve kit with a minor modification. The call rate and concordance of genotypes from each sample were compared to those obtained from whole blood samples extracted by standard PCI extraction.</p> <p>Findings</p> <p>An ANOVA analysis indicated no significant difference (P > 0.72) in BovineSNP50 genotype call rate between DNA extracted from FTA cards by the GenSolve kit or extracted from whole blood by PCI. Two sample t-tests demonstrated that the DNA extracted from the FTA cards produced genotype call and concordance rates that were not different to those produced by assaying DNA samples extracted by PCI from whole blood.</p> <p>Conclusion</p> <p>We conclude that DNA extracted from FTA cards by the GenSolve kit is of sufficiently high quality to produce results comparable to those obtained from DNA extracted from whole blood when assayed by the Illumina iSelect technology. Additionally, we validate the use of nasal swabs as an alternative to venous blood or buccal samples from animal subjects for reliably producing high quality genotypes on this platform.</p

Differential gene expression in multiple neurological, inflammatory and connective tissue pathways in a spontaneous model of human small vessel stroke

Aims: Cerebral small vessel disease (SVD) causes a fifth of all strokes plus diffuse brain damage leading to cognitive decline, physical disabilities and dementia. The aetiology and pathogenesis of SVD are unknown, but largely attributed to hypertension or microatheroma. Methods: We used the spontaneously hypertensive stroke-prone rat (SHRSP), the closest spontaneous experimental model of human SVD, and age-matched control rats kept under identical, non-salt-loaded conditions, to perform a blinded analysis of mRNA microarray, qRT-PCRand pathway analysis in two brain regions (frontal and midcoronal) commonly affected by SVD in the SHRSP at age five, 16 and 21 weeks. Results: We found gene expression abnormalities, with fold changes ranging from 2.5 to 59 for the 10 most differentially expressed genes, related to endothelial tight junctions (reduced), nitric oxide bioavailability (reduced), myelination (impaired), glial and microglial activity (increased), matrix proteins (impaired), vascular reactivity (impaired) and albumin (reduced), consistent with protein expression defects in the same rats. All were present at age 5 weeks thus pre-dating blood pressure elevation. ‘Neurological’ and ‘inflammatory’ pathways were more affected than ‘vascular’ functional pathways. Conclusions: This set of defects, although individually modest, when acting in combination could explain the SHRSP's susceptibility to microvascular and brain injury, compared with control rats. Similar combined, individually modest, but multiple neurovascular unit defects, could explain susceptibility to spontaneous human SVD

Measurement of a Magnetic Field in a Leading Arm High Velocity Cloud

Using a recent catalogue of extragalactic Faraday rotation derived from the NRAO VLA Sky Survey we have found an agreement between Faraday rotation structure and the HI emission structure of a High Velocity Cloud (HVC) associated with the Leading Arm of the Magellanic System. We suggest that this morphological agreement is indicative of Faraday rotation through the HVC. Under this assumption we have used 48 rotation measures through the HVC, together with estimates of the electron column density from H-\alpha\ measurements and QSO absorption lines to estimate a strength for the line-of-sight component of the coherent magnetic field in the HVC of > 6 {\rm \mu G}$. A coherent magnetic field of this strength is more than sufficient to dynamically stabilize the cloud against ram pressure stripping by the Milky Way halo and may also provide thermal insulation for the cold cloud. We estimate an upper limit to the ratio of random to coherent magnetic field of$B_{r}/B_{||} < 0.8$, which suggests that the random field does not dominate over the coherent field as it does in the Magellanic Clouds from which this HVC likely originates.Comment: 17 pages, 3 figure

Chemical Measurement and Fluctuation Scaling

Main abstract: Fluctuation scaling reports on all processes producing a data set. Some fluctuation scaling relationships, such as the Horwitz curve, follow exponential dispersion models which have useful properties. The mean-variance method applied to Poisson distributed data is a special case of these properties allowing the gain of a system to be measured. Here, a general method is described for investigating gain (G), dispersion (β), and process (α) in any system whose fluctuation scaling follows a simple exponential dispersion model, a segmented exponential dispersion model, or complex scaling following such a model locally. When gain and dispersion cannot be obtained directly, relative parameters, GR and βR, may be used. The method was demonstrated on data sets conforming to simple, segmented, and complex scaling. These included mass, fluorescence intensity, and absorbance measurements and specifications for classes of calibration weights. Changes in gain, dispersion, and process were observed in the scaling of these data sets in response to instrument parameters, photon fluxes, mathematical processing, and calibration weight class. The process parameter which limits the type of statistical process that can be invoked to explain a data set typically exhibited 04 possible. With two exceptions, calibration weight class definitions only affected β. Adjusting photomultiplier voltage while measuring fluorescence intensity changed all three parameters (0<α<0.8; 0<βR<3; 0<GR<4.1). The method provides a framework for calibrating and interpreting uncertainty in chemical measurement allowing robust compar ison of specific instruments, conditions, and methods. Supporting information abstract: On first inspection, fluctuation scaling data may appear to approximate a straight line when log transformed. The data presented in figure 5 of the main text gives a reasonable approximation to a straight line and for many purposes this would be sufficient. The purpose of the study of fluorescence intensity was to determine whether adjusting the voltage of a photomultiplier tube while measuring a fluorescent sample changes the process (α), the dispersion (β) and/or the gain (G). In this regard, the linear model established that PMT setting affects more than the gain. However, a detailed analysis beginning with testing for model mis-specification provides additional information. Specifically, Poisson behavior is only seen over a limited wavelength range in the 600 V and 700 V data sets

Two Large HI Shells in the Outer Galaxy near l=279 degrees

As part of a survey of HI 21-cm emission in the Southern Milky Way, we have detected two large shells in the interstellar neutral hydrogen near l=279 deg. The center velocities are +36 and +59 km/s, which puts the shells at kinematic distances of 7 and 10 kpc. The larger shell is about 610 pc in diameter and very empty, with density contrast of at least 15 between the middle and the shell walls. It has expansion velocity of about 20 km/s and swept up mass of several million solar masses. The energy indicated by the expansion may be as high as 2.4 X 10^53 ergs. We estimate its age to be 15 to 20 million years. The smaller shell has diameter of about 400 pc, expansion velocity about 10 km/s and swept up mass of about 10^6 solar masses. Morphologically both regions appear to be shells, with high density regions mostly surrounding the voids, although the first appears to have channels of low density which connect with the halo above and below the HI layer. They lie on the edge of the Carina arm, which suggests that they may be expanding horizontally into the interarm region as well as vertically out of the disk. If this interpretation is correct, this is the first detection of an HI chimney which has blown out of both sides of the disk.Comment: 21 pages, 14 jpeg figures, accepted for publication in A

The (Re-)Discovery of G350.1-0.3: A Young, Luminous Supernova Remnant and Its Neutron Star

We present an XMM-Newton observation of the long-overlooked radio source G350.1-0.3. The X-ray spectrum of G350.1-0.3 can be fit by a shocked plasma with two components: a high-temperature (1.5 keV) region with a low ionization time scale and enhanced abundances, plus a cooler (0.36 keV) component in ionization equilibrium and with solar abundances. The X-ray spectrum and the presence of non-thermal, polarized, radio emission together demonstrate that G350.1-0.3 is a young, luminous supernova remnant (SNR), for which archival HI and 12-CO data indicate a distance of 4.5 kpc. The diameter of the source then implies an age of only ~900 years. The SNR's distorted appearance, small size and the presence of 12-CO emission along the SNR's eastern edge all indicate that the source is interacting with a complicated distribution of dense ambient material. An unresolved X-ray source, XMMU J172054.5-372652, is detected a few arcminutes west of the brightest SNR emission. The thermal X-ray spectrum and lack of any multi-wavelength counterpart suggest that this source is a neutron star associated with G350.1-0.3, most likely a "central compact object", as seen coincident with other young SNRs such as Cassiopeia A.Comment: 6 pages, uses emulateapj. One B/W figure, one color figure. Minor text changes and update to Fig 2 following referee's report. ApJ Letters, in pres

The Southern Galactic Plane Survey: The Test Region

The Southern Galactic Plane Survey (SGPS) is a project to image the HI line emission and 1.4 GHz continuum in the fourth quadrant of the Milky Way at high resolution using the Australia Telescope Compact Array (ATCA) and the Parkes Radio Telescope. In this paper we describe the survey details and goals, present lambda 21-cm continuum data, and discuss HI absorption and emission characteristics of the SGPS Test Region (325.5 deg < l < 333.5 deg; -0.5 deg < b < +3.5 deg). We explore the effects of massive stars on the interstellar medium (ISM) through a study of HI shells and the HI environments of HII regions and supernova remnants. We find an HI shell surrounding the HII region RCW 94 which indicates that the region is embedded in a molecular cloud. We give lower limits for the kinematic distances to SNRs G327.4+0.4 and G330.2+1.0 of 4.3 kpc and 4.9 kpc, respectively. We find evidence of interaction with the surrounding HI for both of these remnants. We also present images of a possible new SNR G328.6-0.0. Additionally, we have discovered two small HI shells with no counterparts in continuum emission.Comment: 17 pages, 7 embedded EPS figures, 10 low-res jpeg figures, uses emulateapj5.sty. Accepted for publication in the Astrophysical Journal. Version with all full resolution figures embedded is available at http://www.astro.umn.edu/~naomi/sgps/papers/SGPS.ps.g

Return to return point memory

We describe a new class of systems exhibiting return point memory (RPM) that are different from those discussed before in the context of ferromagnets. We show numerically that one dimensional random Ising antiferromagnets have RPM, when configurations evolve from a large field. However, RPM is violated when started from some stable configurations at finite field unlike in the ferromagnetic case. This implies that the standard approach to understanding ferromagnetic RPM systems will fail for this case. We also demonstrate RPM with a set of variables that keep track of spin flips at each site. Conventional RPM for the spin configuration is a projection of this result, suggesting that spin flip variables might be a more fundamental representation of the dynamics. We also present a mapping that embeds the antiferromagnetic chain in a two dimensional ferromagnetic model, and prove RPM for spin exchange dynamics in the interior of the chain with this mapping