Apolipoprotein E delivery by peritoneal implantation of encapsulated recombinant cells improves the hyperlipidaemic profile in apoE-deficient mice

Plasma apolipoprotein E (apoE) is a 34-kDa polymorphic protein which has atheroprotective actions by clearing remnant lipoproteins and sequestering excess cellular cholesterol. Low or dysfunctional apoE is a risk factor for hyperlipidaemia and atherosclerosis, and for restenosis after angioplasty. Here, in short-term studies designed to establish proof-of-principle, we investigate whether encapsulated recombinant Chinese hamster ovary (CHO) cells can secrete wild-type apoE3 protein in vitro and then determine whether peritoneal implantation of the microcapsules into apoE-deficient (apoE(-/-)) mice reduces their hypercholesterolaemia.Recombinant CHO-E3 cells were encapsulated into either alginate poly-L-lysine or alginate polyethyleneimine/polybrene microspheres. After verifying stability and apoE3 secretion, the beads were then implanted into the peritoneal cavity of apoE(-/-) mice; levels of plasma apoE3, cholesterol and lipoproteins were monitored for up to 14 days post-implantation.Encapsulated CHO-E3 cells continued to secrete apoE3 protein throughout a 60-day study period in vitro, though levels declined after 14 days. This cell-derived apoE3 was biologically active. When conditioned medium from encapsulated CHO-E3 cells was incubated with cultured cells pre-labelled with [H-3]-cholesterol, efflux of cholesterol was two to four times greater than with normal medium (at 8 h, for example, 7.4+/-0.3% vs. 2.4+/-0.2% of cellular cholesterol; P<0.001). Moreover, when secreted apoE3 was injected intraperitoneally into apoE(-/-) mice, apoE3 was detected in plasma and the hyperlipidaemia improved. Similarly, when alginate polyethyleneimine/polybrene capsules were implanted into the peritoneum of apoE(-/-) mice, apoE3 was secreted into plasma and at 7 days total cholesterol was reduced, while atheroprotective high-density lipoprotein (HDL) increased. In a second study, apoE was detectable in plasma of five mice treated with alginate poly-L-lysine beads, 4 and 7 days post-implantation, though not at day 14. Furthermore, their hypercholesterolaemia was reduced, while HDL was clearly elevated in all mice at days 4 and 7 (from 18.4+/-6.2% of total lipoproteins to 31.1+/-6.8% at 7 days; P<0.001); however, these had rebounded by day 14, possibly due to the emergence of anti-apoE antibodies.We conclude that microencapsulated apoE-secreting cells have the potential to ameliorate the hyperlipidaemia of apoE deficiency, but that the technology must be improved to become a feasible therapeutic to treat atherosclerosis. (C) 2004 Elsevier B.V. All rights reserved

Thinking Like a Lawyer, Designing Like an Architect: Preparing Students for the 21st Century Practice

Various law schools—Chicago-Kent Law School, New York Law School, Vermont Law School, and Georgetown Law Center among them—are beginning to offer innovative classes in which students learn to build legal expert systems intended to enhance access to the legal system. Working in platforms that do not require technical expertise, students are able to build apps that incorporate rules-based logic, factor balancing, and mathematical operations to implement the reasoning of a regulatory regime. In this essay, we suggest that teaching students to design apps furthers pedagogic goals associated with the traditional law school curriculum and clinical teaching. In designing legal expert systems, students are required to engage in careful legal analysis and anticipate the problems and questions a typical user will have. Students also need to learn to communicate legal concepts and categories in precise and plain language. Contrary to the traditional law school curriculum, however, which emphasizes case-by-case analysis, in clinics that focus on building legal expert systems, students learn to develop systemic solutions to legal problems. By exposing students to principle of systems design, these classes prepare them for the emerging challenges of 21st century practice

Thinking Like a Lawyer, Designing Like an Architect: Preparing Students for the 21st Century Practice

Various law schools—Chicago-Kent Law School, New York Law School, Vermont Law School, and Georgetown Law Center among them—are beginning to offer innovative classes in which students learn to build legal expert systems intended to enhance access to the legal system. Working in platforms that do not require technical expertise, students are able to build apps that incorporate rules-based logic, factor balancing, and mathematical operations to implement the reasoning of a regulatory regime. In this essay, we suggest that teaching students to design apps furthers pedagogic goals associated with the traditional law school curriculum and clinical teaching. In designing legal expert systems, students are required to engage in careful legal analysis and anticipate the problems and questions a typical user will have. Students also need to learn to communicate legal concepts and categories in precise and plain language. Contrary to the traditional law school curriculum, however, which emphasizes case-by-case analysis, in clinics that focus on building legal expert systems, students learn to develop systemic solutions to legal problems. By exposing students to principle of systems design, these classes prepare them for the emerging challenges of 21st century practice

Resolved magnetic structures in the disk-halo interface of NGC 628

Magnetic fields are essential to fully understand the interstellar medium (ISM) and its role in the disk-halo interface of galaxies is still poorly understood. Star formation is known to expel hot gas vertically into the halo and these outflows have important consequences for mean-field dynamo theory in that they can be efficient in removing magnetic helicity. We perform new observations of the nearby face-on spiral galaxy NGC 628 with the Karl G. Jansky Very Large Array (JVLA) at S-band and the Effelsberg 100-m telescope at frequencies of 2.6 GHz and 8.35 GHz. We obtain some of the most sensitive radio continuum images in both total and linearly polarised intensity of any external galaxy observed so far in addition to high-quality images of Faraday depth and polarisation angle from which we obtained evidence for drivers of magnetic turbulence in the disk-halo connection. Such drivers include a superbubble detected via a significant Faraday depth gradient coinciding with a HI hole. We observe an azimuthal periodic pattern in Faraday depth with a pattern wavelength of 3.7$\pm$ 0.1 kpc, indicating Parker instabilities. The lack of a significant anti-correlation between Faraday depth and magnetic pitch angle indicates that these loops are vertical in nature with little helical twisting, unlike in IC 342. We find that the magnetic pitch angle is systematically larger than the morphological pitch angle of the polarisation arms which gives evidence for the action of a large-scale dynamo where the regular magnetic field is not coupled to the gas flow and obtains a significant radial component. We additionally discover a lone region of ordered magnetic field to the north of the galaxy with a high degree of polarisation and a small pitch angle, a feature that has not been observed in any other galaxy so far and is possibly caused by an asymmetric HI hole.Comment: 25 pages, Accepted for publication in Astronomy and Astrophysic

In the Interests of clients or commerce? Legal aid, supply, demand, and 'ethical indeterminacy' in criminal defence work

As a professional, a lawyer's first duty is to serve the client's best interests, before simple monetary gain. In criminal defence work, this duty has been questioned in the debate about the causes of growth in legal aid spending: is it driven by lawyers (suppliers) inducing unnecessary demand for their services or are they merely responding to increased demand? Research reported here found clear evidence of a change in the handling of cases in response to new payment structures, though in ways unexpected by the policy's proponents. The paper develops the concept of 'ethical indeterminacy' as a way of understanding how defence lawyers seek to reconcile the interests of commerce and clients. Ethical indeterminacy suggests that where different courses of action could each be said to benefit the client, the lawyer will tend to advise the client to decide in the lawyer's own interests. Ethical indeterminacy is mediated by a range of competing conceptions of 'quality' and 'need'. The paper goes on to question the very distinction between 'supply' and 'demand' in the provision of legal services

Radio haloes in nearby galaxies modelled with 1D cosmic-ray transport using SPINNAKER

We present radio continuum maps of 12 nearby ($D\leq 27~\rm Mpc$), edge-on ($i\geq 76^{\circ}$), late-type spiral galaxies mostly at $1.4$ and 5 GHz, observed with the Australia Telescope Compact Array, Very Large Array, Westerbork Synthesis Radio Telescope, Effelsberg 100-m and Parkes 64-m telescopes. All galaxies show clear evidence of radio haloes, including the first detection in the Magellanic-type galaxy NGC 55. In 11 galaxies, we find a thin and a thick disc that can be better fitted by exponential rather than Gaussian functions. We fit our SPINNAKER (SPectral INdex Numerical Analysis of K(c)osmic-ray Electron Radio-emission) 1D cosmic-ray transport models to the vertical model profiles of the non-thermal intensity and to the non-thermal radio spectral index in the halo. We simultaneously fit for the advection speed (or diffusion coefficient) and magnetic field scale height. In the thick disc, the magnetic field scale heights range from 2 to 8 kpc with an average across the sample of $3.0\pm 1.7~\rm kpc$; they show no correlation with either star-formation rate (SFR), SFR surface density ($\Sigma_{\rm SFR}$) or rotation speed ($V_{\rm rot}$). The advection speeds range from 100 to $700~\rm km\,s^{-1}$ and display correlations of $V\propto \rm SFR^{0.36\pm 0.06}$ and $V\propto \Sigma_{\rm SFR}^{0.39\pm 0.09}$; they agree remarkably well with the escape velocities ($0.5\leq V/V_{\rm esc}\leq 2$), which can be explained by cosmic-ray driven winds. Radio haloes show the presence of disc winds in galaxies with $\Sigma_{\rm SFR} > 10^{-3}~\rm M_{\odot}\,yr^{-1}\,kpc^{-2}$ that extend over several kpc and are driven by processes related to the distributed star formation in the disc.Comment: 39 pages, 20 colour figures, 10 tables. Accepted by MNRA

Carey Young’s 'Palais de Justice'

The symposium for this issue comprises six responses to the video artwork Palais de Justice (2017) by artist Carey Young. The video presents a study of the life of Brussels’ vast, late-nineteenth-century court building. In Palais de Justice, Young presents ‘a legal system seemingly centered on, and perhaps controlled by women’. The respondents are Jeanne Gaakeer, Ruth Herz, Joan Kee, Linda Mulcahy, Jeremy Pilcher and Gary Watt. Jeanne Gaakeer and Ruth Herz have the distinction of being, not only internationally respected scholars, but also experienced judges. Jeanne Gaakeer is a judge practicing in the Netherlands and Ruth Herz was formerly a judge in Germany. The six responses are followed by the artist’s own reflections on her artwork and her response to the commentators’ responses. Joan Kee writes that ‘Young highlights access as a key entry point for thinking about the law. Who can avail themselves of the law? Who may enter (or exit) the courts? Who is excluded and by whose authority? The surreptitious looking and peering that define the experience of watching the film suggests how these questions deny ready answers’

The effects of collagen concentration and crosslink density on the biological, structural and mechanical properties of collagen-GAG scaffolds for bone tissue engineering.

In this study, we examined the effects of varying collagen concentration and crosslink density on the biological, structural and mechanical properties of collagen-GAG scaffolds for bone tissue engineering. Three different collagen contents (0.25%, 0.5% and 1% collagen) and two different dehydrothermal (DHT) crosslinking processes [1] 105 degrees C for 24 h and [2] 150 degrees C for 48 h were investigated. These scaffolds were assessed for (1) pore size, (2) permeability (3) compressive strength and (4) cell viability. The largest pore size, permeability rate, compressive modulus, cell number and cell metabolic activity was all found to occur on the 1% collagen scaffold due to its increased collagen composition and the DHT treatment at 150 degrees C was found to significantly improve the mechanical properties and not to affect cellular number or metabolic activity. These results indicate that doubling the collagen content to 1% and dehydrothermally crosslinking the scaffold at 150 degrees C for 48 h has enhanced mechanical and biological properties of the scaffold making it highly attractive for use in bone tissue engineering