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There are few nationwide descriptive studies of longitudinal drug use and residual cardiovascular risk in patients with myocardial infarction (MI) in contemporary clinical practice. The objectives of this work were to describe characteristics and longitudinal cardiovascular drug use of patients with a first acute MI in Norway, and to quantify residual risks of cardiovascular events and death. Using nationwide health registries in Norway, we identified 43 750 adults with a first MI (2010 to 2015) and ≥1 prescription for antiplatelet medication. We described cardiovascular medication post-MI and calculated residual cardiovascular risks. Between 3 months and 13–15 months post MI, medication use dropped from 93.3% to 75.1% for low-dose aspirin, 78.1% to 11.0% for dual antiplatelet therapy, 91.6% to 78.7% for antihypertensives, and 88.0% to 70.7% for lipid-lowering therapy. Incidence rate ratios (IRRs) for recurrent MI were similar between subpopulations at 12 months and notably different at 12–36 months. IRRs (95% CIs) at 12–36 months were 1.52 (1.26–1.82) for 65–74 years, 2.26 (1.88–2.71) for 75–84 years, and 3.97 (3.29–4.79) for ≥85 years (vs. 18–49 years), 2.42 (2.18–2.69) for those with ischaemic heart disease (IHD), 2.26 (1.97–2.59) for peripheral artery disease (PAD), 2.17 (1.98–2.36) for hypertension, and 1.82 (1.65–2.01) for diabetes. In conclusion, secondary prevention medication use 13–15 months following a first MI is suboptimal among patients in Norway. The elderly and those with IHD, PAD, diabetes, or hypertension are at high-risk for recurrent MI/stroke/death and should be managed closely beyond the first year.

In 2018, the City of Toronto’s municipal election overlapped with a provincial election that brought a new government to office. While the municipal election ran for a protracted period from May 1 to October 22, the provincial election began on May 9 and ended about four weeks later, on June 7.1 On July 27, after only a few weeks in office, the provincial government tabled the Better Local Government Act (BLGA) and proclaimed the Bill into law on August 14.2 The BLGA reduced Toronto City Council from 47 to 25 wards and reset the electoral process, mandating that the election proceed under a different concept of representation for City Council.3

In 1985, it was largely unknown how the Supreme Court of Canada would respond to the Charter.1 At first glance, a drugstore’s right to be open for business on Sunday, selling groceries, plastic cups, and a bicycle lock, seemed an unlikely source of inspiration for the Court’s first pronouncement on the essence of freedom. Perhaps unexpectedly, the justices enforced the entitlement, finding that a Sunday closing law compelling a corporation to comply with the Christian Sabbath infringed section 2(a)’s guarantee of religious freedom.2 In doing so, R v Big M Drug Mart defined freedom as “the absence of coercion or constraint,” stating without equivocation that no one who is compelled “to a course of action or inaction” is “truly free”.3 In Justice Dickson’s considered view, coercion includes “blatant forms of compulsion”, such as “direct commands to act or refrain from acting on pain of sanctions”, as well as forms of indirect control.4 In plain and unmistakeable terms, Big M promised that, under the Charter, “no one is to be forced to act in a way contrary to his beliefs or conscience”.5 * Professor Emeritus, Osgoode Hall Law School. I thank Kate Bezanson and Alison Braley-Rattai for includingme in this special issue of Constitutional Forum, and am grateful to Kate Bezanson for her comments onan earlier draft. I also thank Ryan Ng (JD 2021) for his valuable research assistance in the preparation ofthis paper. Finally, I note that I was a member of York University’s Free Speech Working Group in fall 2018.This paper does not in any way express the views of York University or the Working Group, which has longsince disbanded. 1Canadian Charter of Rights and Freedoms, s 2(a), Part I of the Constitution Act, 1982, being Schedule B of the Canada Act 1982 (UK), 1982, c 11 [Charter].2R v Big M Drug Mart, [1985] 1 SCR 295, 18 DLR (4th) 321 [Big M].3Ibid at 336.4Ibid.5 Ibid at 337.

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This Review Essay forms part of a special issue of the Manitoba Law Journal (forthcoming) dedicated to Richard Albert, Constitutional Amendments: Making, Breaking, and Changing Constitutions. Years in the making, Constitutional Amendments explains how amendment rules define a constitution’s integrity, ensuring its longevity by allowing and even inviting formal changes to its text. Albert’s work is prodigious and monumental, connecting abstract issues of textual design to the follies of constitutional amendment over diverse variables of time and place. This Review focuses selectively on Constitutional Amendments and its implications for Canadian amendment constitutionalism, exploring Albert’s view that amendment rules expose a constitution’s “deepest vulnerabilities” and reveal its “greatest strengths”. Specifically, the Review draws on Albert’s work to suggest a simple but sharp insight linking the 1867 Constitutions’s failure to provide textual amendment rules to the steadfast unamendability of the Canadian Constitution today.

Health care organizations are obligated to provide safe and effective treatment to their patients and also protect the safety of their workers. This paper analyzes the tensions arising from legislative regimes that, respectively, protect privacy and workplace safety, using a large, tertiary high-secure forensic psychiatric hospital in Ontario, Canada, as an example. In Ontario, the Personal Health Information Protection Act (PHIPA) prohibits personal health information (PHI) from being disclosed to individuals who fall outside the "circle of care," including nonclinical employees who have direct involvement with patients and may be at risk of violence. PHIPA permits the disclosure of information where there is a risk of violence, but the statute's scheme for privacy protection was not designed to address, and may not be compatible with, the operations and requirements of high-secure forensic and other psychiatric hospitals. At the same time, the Occupational Health and Safety Act (OHSA) creates a regulatory framework that sets health and safety standards, including an employer's duty to disclose the risk of violence. OHSA prosecutions and proceedings demonstrate how these duties have been enforced against psychiatric hospitals. We examine this regulatory backdrop, explaining that PHIPA provides little guidance to psychiatric hospitals, where the risk of violence is elevated. We also discuss issues of dual compliance that arise from a hospital's legal obligations under PHIPA and OHSA. Finally, we turn to the ongoing clinical and operational challenges, suggesting strategies for increasing staff safety. These include strengthening the therapeutic alliance and providing patients with the option of consenting to disclosure of PHI to those outside the circle of care.

Hafting adhesives play a crucial role in the study of composite objects such as hunting weapons. However, they tend to degrade rapidly and, when they exceptionally preserve, they represent only micro remains. Over the last 20 years, different techniques have been applied to study such hafting residues. These techniques include optical microscopy, scanning electron microscopy and gas chromatography / mass spectrometry. Hafting adhesives have also been radiocarbon dated in some cases. Here, we present a multi-analytical approach using these techniques as well as photogrammetry and FTIR spectroscopy. Our integrated approach, combining these six techniques in a strategic order, allows us to document, characterise chemically and date hafting adhesives. We applied this methodology to a set of stone tools discovered in Sart-Saint-Laurent (Belgium) without any archaeological context. This multi-analytical work revealed that several of the tools have been hafted with a heated Pinaceae resin around the 2nd-4th century CE, although some of the artefacts can be attributed to the Neolithic. More generally, this work demonstrates the complementary nature of all the techniques employed and their crucial contribution to fully elucidate hafting techniques.

The performance of a field-enhanced emitter (FEE) array in an enclosed and ex-situ operated device has long been known to deviate from the ideal characteristics observed in laboratory settings. Here we report on the performance and failure mechanisms measured in FEE arrays operating in high-voltage sealed x-ray generators. The emitters are fabricated from etched silicon forming pyramids which are coated with metal. Unlike many previously reported arrays, here the pitch is large (1cm) and affords easy measurement and diagnosis of individual emitters. We report on emitter performance and likely causes of deviations from ideal, including the impact of anode treatment, emitter geometry and catastrophic failure mechanisms.

Part of a collection of papers on section 2’s “forgotten freedoms” (forthcoming (2020), 100 S.C.L.R.(2d)), “Big M’s Forgotten Legacy of Freedom” returns to the Supreme Court of Canada’s foundational concept of freedom as the absence of coercion or constraint. That early and important legacy of freedom under the Charter failed to inspire and influence the evolution of section 2, especially section 2(b)’s guarantee of expressive freedom and section 2(d)’s guarantee of associational freedom. This paper both claims and demonstrates that section 2’s fundamental freedoms have been less meaningful as a result. In doing so, it places emphasis on section 2(b) and its legacy of limits, rather than freedom, and section 2(d) and its legacy of relative failure, except – and only recently – for labour union claims. After more than thirty-five years, the section 2 jurisprudence lacks a fundamental conception of freedom and theory of section 2 constitutionalism. Big M provides a starting point in working toward those goals.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) binds to the low density lipoprotein receptor (LDLR) at the cell surface and is internalized as a complex with the LDLR. In the acidic milieu of the sorting endosome, PCSK9 remains bound to the LDLR and prevents the LDLR from folding over itself to adopt a closed conformation. As a consequence, the LDLR fails to recycle back to the cell membrane. Even though it is the catalytic domain of PCSK9 that interacts with the LDLR at the cell surface, the structurally disordered segment consisting of residues 31-60 and which is rich in acidic residues, has a negative effect both on autocatalytic cleavage and on the activity of PCSK9 towards the LDLR. Thus, this unstructured segment represents an autoinhibitory domain of PCSK9. One may speculate that post-translational modifications within residues 31-60 may affect the inhibitory activity of this segment, and represent a mechanism for fine-tuning the activity of PCSK9 towards the LDLR. Our data indicate that the inhibitory effect of this unstructured segment results from an interaction with basic residues of the catalytic domain of PCSK9. Mutations in the catalytic domain which involve charged residues, could therefore be gain-of-function mutations by affecting the positioning of this segment.

Mesa is a highly scalable analytic data warehousing system that stores critical measurement data related to Google's Internet advertising business. Mesa is designed to satisfy a complex and challenging set of user and systems requirements, including near real-time data ingestion and retrieval, as well as high availability, reliability, fault tolerance, and scalability for large data and query volumes. Specifically, Mesa handles petabytes of data, processes millions of row updates per second, and serves billions of queries that fetch trillions of rows per day. Mesa is geo-replicated across multiple datacenters and provides consistent and repeatable query answers at low latency, even when an entire datacenter fails. This paper presents the Mesa system and reports the performance and scale that it achieves.

The need for a low-cost, low-dose 3D imaging capability, as an alternative to CT and MRI scanners, has driven the development of a multi-cathode, stationary Digital Tomosynthesis (s-DT) scanner. Demonstration experiments with an array of cold cathodes as electron sources have been reported by multiple groups. We report the operation of a protoype with a different field emitter composition and geometry, at 60kV, to obtain images of a dental phantom, and the subsequent reconstruction of the raw images to obtain 3D slices. Thermal heating of the emitters was found to be the primary factor limiting performance of the device, and several well-established control mechanisms were employed to mitigate this. These include using a ballast resistance, metal coatings, conditioning, and optimizing the duty cycle at operating voltage. We report on the consequent improvement in device flux, lifetime and yield, and present the future outlook of our device.

Freedom of expression and freedom of association are guaranteed by section 2 of the Charter of Rights and Freedoms. These freedoms are closely related, conceptually and philosophically, but evolved in different directions under the Charter. Whereas section 2(b)’s guarantee of expressive freedom generated a rich jurisprudence across diverse issues, section 2(d)’s attention focused on associational freedom in the context of labour union activities. The authors draw on a pocket of section 2(b) case law on picketing and other labour-related expressive activities to bring these guarantees into comparison. In doing so, they comment on the Supreme Court of Canada’s interpretation of each guarantee, including the constitutionalization of key aspects of labour relations under section 2(d). In addition, the authors critique the Court’s jurisprudence, emphasizing the central importance of protecting protest and dissent activities under both guarantees.

The need for a low-cost, low-dose 3D imaging capability, as an alternative to CT and MRI scanners, has driven the development of a multi-cathode, stationary Digital Tomosynthesis (s-DT) scanner. Demonstration experiments with an array of cold cathodes as electron sources have been reported by multiple groups. We report the operation of a device with a different field emitter geometry, at 60kV, to obtain images of a dental phantom, and the subsequent reconstruction of the raw images to obtain 3D slices. Failure mechanisms of the emitters, possible mitigation options, and the future outlook of emitter array devices are also discussed.

This paper presents the results of a scientific investigation on a preserved head of hair, discovered in 1839 inside a lead coffin beneath the Norman Abbey of Romsey (Hampshire, UK). Accelerator mass spectrometry (AMS) radiocarbon dating, modelled to account for dietary marine protein intake indicated by stable isotope analysis (δ 13 C and δ 15 N), suggests that this individual died between 895–1123 calAD (95.4% probability). A wooden ‘pillow’ found supporting the hair produced a date of 895–1016 calAD (95.4% probability). These results suggest that this burial dates from the end of the ninth century to the early twelfth century, placing it in the late Saxon or immediately post-Conquest era. Analysis by Gas Chromatography/Mass Spectrometry (GC/MS) of residue adhering to the hair surface indicates the presence of Pinacea resin, deriving either from funerary ritual or hair care during life.

Mesa is a highly scalable analytic data warehousing system that stores critical measurement data related to Google's Internet advertising business. Mesa is designed to satisfy a complex and challenging set of user and systems requirements, including near real-time data ingestion and queryability, as well as high availability, reliability, fault tolerance, and scalability for large data and query volumes. Specifically, Mesa handles petabytes of data, processes millions of row updates per second, and serves billions of queries that fetch trillions of rows per day. Mesa is geo-replicated across multiple datacenters and provides consistent and repeatable query answers at low latency, even when an entire datacenter fails. This paper presents the Mesa system and reports the performance and scale that it achieves.

It is accepted that Canada’s Constitution is almost impossible to amend, and that this amendment rigidity stems from the Constitution’s patriation in 1982 and two failed reform initiatives, the Meech Lake and Charlottetown Accords. The high-stakes drama of 1982, threat of Quebec separation and denouement of the post-patriation Accords were events of singular urgency which have dominated the literature and consigned the longer history of amendment to the background. This article provides a corrective which explores Canada’s two uneven periods of constitutional change – before and after textual rules – and in doing so theorizes that rich insights into the riddles of Canada’s amendment constitutionalism are found in the interface between the legality or formality of amendment, and its legitimacy or acceptance. The article explains how the legality and legitimacy of amendment failed to align prior to patriation – when amendment was governed by a concept of statutory legality – and after patriation, when rules of constitutional legality were adopted. Though the textual amendment rules are admittedly rigid, the role legitimacy plays in Canada’s amendment rigidity cannot be overlooked. As Canada approaches its sesquicentennial anniversary in 2017 there is lingering uncertainty on what is required to align the legitimacy of constitutional change with the formal criteria of amendment legality.

“Collateral Thoughts” is part of a special issue edited by Professor James Allan, who invited and challenged a group of scholars to select and discuss a favourite law review article. I chose “The Charter Dialogue Between Courts and Legislatures” because it is the most influential article to date in the Charter of Rights and Freedom’s relatively short history (since 1982). I call this reflection “Collateral Thoughts” because my interest is less in the merits of dialogue theory than in its remarkable impact, at home in Canada as well as abroad, in the broader reaches of comparative constitutionalism and constitutional theory. In the main, this reflection asks how and why “dialogue” became a runaway concept, and considers what that tells us about the nature and formation of constitutional theory. It shows that Dialogue was fundamentally connected to Canada’s catharsis of rights, in 1982 and in the early years of Charter interpretation. That is why its claim that the legitimacy battles which define American judicial review are irrelevant – because Canada’s system of rights protection is based on “dialogue” – was so explosive. Not only did this article command attention in Canada, the concept of dialogue would be “reified”, castigated and deeply analyzed by scholars and Commonwealth jurisdictions who wondered whether weaker or weak-form judicial review was institutionally possible. Much like other theories in the US constitutional tradition, Dialogue responded to controversy by theorizing and attempting to legitimize review. Despite failing in its objective to eliminate legitimacy concerns about review, Dialogue catalyzed a national and international movement in constitutional thought.

Thomas Healy’s The Great Dissent re-treads the familiar story of US Supreme Court Justice Holmes’s First Amendment conversion between March and November 1919, when he launched his marketplace of ideas theory and strong-form version of the clear and present danger doctrine. Healy’s book demonstrates that fresh perspectives on this vital and ever-intriguing change of mind or transformation on Holmes’s part remain possible. The review offers its own perspective by highlighting the process of “reverse mentoring” which took place, in which the older jurist was mentored on free speech issues by the emerging thought leaders of the day – Laski, Frankfurter, Chafee – and showing how Justice Holmes’s landmark dissent in Abrams was nonetheless and indisputably a product of his own jurisprudential ingenuity.

This Article was written for Osgoode Hall Law School’s annual Constitutional Cases conference, and provides the keynote overview of the McLachlin Court’s 2014 constitutional jurisprudence. The Court’s 2014 constitutional decisions (Appointment and Senate References; Tsilqot’in Nation; Trial Lawyers) and restrictions on Mr. Big operations (Hart), in combination with a tsunami of Charter decisions early in 2015 (the 2015 Labour Trilogy; Carter v. Canada; R. v. Nur; and others), made this a legacy-building year. More than an overview, this Article probes the nature of the McLachlin Court’s legacy this year and the relationship between legal and political dynamics, to ask: in light of Chief Justice McLachlin’s modest goals as leader of the Court and the immediate backdrop of the Nadon controversy, the political leadership’s attack on the Chief Justice, and poor relations between the institutions, should the jurisprudence of this period be seen as a passive response to events or a more active assertion of judicial authority? Whether this legacy-making jurisprudence was a function of serendipity or opportunity (and perhaps both) is an intriguing question that sheds light on the Chief Justice and her Court in what was a singular and unforgettable year on legal and political measures.

In this study we have characterized a naturally occurring truncated form of the low density lipoprotein receptor (LDLR). Western blot analysis of transfected cells indicated that the truncated form (∆N-LDLR) is a degradation product of the full-length LDLR generated by cleavage in the linker region between ligand-binding repeats 4 and 5 of the ligand-binding domain. The cleavage of the linker was not caused by components of the culture media, as heat inactivation of the media did not prevent cleavage. Rather, it is assumed that cleavage was caused by an enzyme secreted from the cells. Biotinylation experiments showed that ∆N-LDLR is located on the cell surface and is detectable approximately 5 h after synthesis of the full-length LDLR. Flow cytometric analysis showed that ∆N-LDLR was not able to bind and internalize low density lipoprotein (LDL). ∆N-LDLR appeared to be equally stable as the full-length LDLR. Thus, generation of ∆N-LDLR does not appear to be the first signal for degradation of the LDLR. The existence of two functionally different populations of LDLRs on the cell surface, of which ∆N-LDLR constitutes 28%, must be taken into account when interpreting results of experiments to study LDLRs on the cell surface. Furthermore, if the cleavage of the linker between ligand-binding repeats 4 and 5 could be prevented by an enzyme inhibitor, this could represent a novel therapeutic strategy to increase the number of functioning LDLRs and thereby decrease the levels of plasma LDL cholesterol.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) binds to the epidermal growth factor homology domain repeat A of the low-density lipoprotein receptor (LDLR) at the cell surface and disrupts recycling of the internalized LDLR. As a consequence, the LDLR is rerouted to the lysosomes for degradation. Although PCSK9 may bind to an LDLR lacking the ligand-binding domain, at least three ligand-binding repeats of the ligand-binding domain are required for PCSK9 to reroute the LDLR to the lysosomes. In this study, we have studied the binding of PCSK9 to an LDLR with or without the ligand-binding domain at increasingly acidic conditions in order to mimic the milieu of the LDLR:PCSK9 complex as it translocates from the cell membrane to the sorting endosomes. These studies have shown that PCSK9 is rapidly released from an LDLR lacking the ligand-binding domain at pH in the range of 6.9-6.1. A similar pattern of release at acidic pH was also observed for the binding to the normal LDLR of mutant PCSK9 lacking the C-terminal domain. Together these data indicate that an interaction between the negatively charged ligand-binding domain of the LDLR and the positively charged C-terminal domain of PCSK9 is required for PCSK9 to remain bound to the LDLR during the early phase of endosomal acidification as the LDLR translocates from the cell membrane to the sorting endosome.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) binds to the low density lipoprotein receptor (LDLR) at the cell surface and disrupts the normal recycling of the LDLR. In this study, we investigated the role of the C-terminal domain for the activity of PCSK9. Experiments in which conserved residues and histidines on the surface of the C-terminal domain were mutated indicated that no specific residues of the C-terminal domain, apart from those responsible for maintaining the overall structure, are required for the activity of PCSK9. Rather, the net charge of the C-terminal domain is important. The more positively charged the C-terminal domain, the higher the activity toward the LDLR. Moreover, replacement of the C-terminal domain with an unrelated protein of comparable size led to significant activity of the chimeric protein. We conclude that the role of the evolutionary, poorly conserved C-terminal domain for the activity of PCSK9 reflects its overall positive charge and size and not the presence of specific residues involved in protein-protein interactions.

The low density lipoprotein receptor (LDLR) binds and internalizes low density lipoprotein (LDL). At the mildly acidic pH of the sorting endosomes, LDL is released from the receptor and the receptor recycles back to the cell membrane. Mutations in the LDLR gene may disrupt the normal function of the LDLR in different ways. Class 5 mutations result in receptors that are able to bind and internalize LDL, but they fail to release LDL in the sorting endosomes and fail to recycle. Instead they are rerouted to the lysosomes for degradation. However, the underlying mechanism remains to be determined. To study the role of the cytoplasmic domain of the LDLR for rerouting Class 5 mutants to the lysosomes, we have performed studies to determine whether Class 5 mutants caused by mutations E387K or V408M are degraded when the cytoplasmic domain has been altered or deleted. As determined by confocal laser-scanning microscopy, these mutant LDLR were inserted into the cell membrane and were able to internalize LDL. As determined by Western blot analysis, Class 5 mutants without a cytoplasmic domain still were degraded after binding LDL. Thus, the cytoplasmic domain does not play a role in rerouting Class 5 mutant LDLR to the lysosomes. Rather, one may speculate that sterical hindrance may prevent Class 5 mutants with bound LDL from entering the narrow recycling tubules of the sorting endosome.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) binds to the low density lipoprotein receptor (LDLR) at the cell surface and mediates intracellular degradation of the LDLR. The amino-terminus of mature PCSK9, residues 31-53 of the prodomain, has an inhibitory effect on this function of PCSK9, but the underlying mechanism is not fully understood. In this study, we have identified two highly conserved negatively charged segments (residues 32-40 and 48-50, respectively) within this part of the prodomain and performed deletions and substitutions to study their importance for degradation of the LDLRs. Deletion of the acidic residues of the longest negatively charged segment increased PCSK9's ability to degrade the LDLR by 31%, whereas a modest 8% increase was observed when these residues were mutated to uncharged amino acids. Thus, both the length and the charge of this part of the prodomain were important for its inhibitory effect. Deletion of the residues of the shorter second negatively charged segment only increased PCSK9's activity by 8%. Substitution of the amino acids of both charged segments to uncharged residues increased PCSK9's activity by 36%. These findings indicate that the inhibitory effect of residues 31-53 of the prodomain is due to the negative charge of this segment. The underlying mechanism could involve the binding of this peptide segment to positively charged structures which are important for PCSK9's activity. One possible candidate could be the histidine-rich C-terminal domain of PCSK9.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) binds to the extracellular domain of the low density lipoprotein receptor (LDLR) at the cell surface, and disrupts the normal recycling of the LDLR. However, the exact mechanism by which the LDLR is re-routed for lysosomal degradation remains to be determined. To clarify the role of the cytoplasmic domain of the LDLR for re-routing to the lysosomes, we have studied the ability of PCSK9 to degrade a chimeric receptor which contains the extracellular and transmembrane domains of the LDLR and the cytoplasmic domain of the transferrin receptor. These studies were performed in CHO T-REx cells stably transfected with a plasmid encoding the chimeric receptor and a novel assay was developed to study the effect of PCSK9 on the LDLR in these cells. Localization, function and stability of the chimeric receptor were similar to that of the wild-type LDLR. The addition of purified gain-of-function mutant D374Y-PCSK9 to the culture medium of stably transfected CHO T-REx cells showed that the chimeric receptor was degraded, albeit to a lower extent than the wild-type LDLR. In addition, a mutant LDLR, which has the three lysines in the intracellular domain substituted with arginines, was also degraded by D374Y-PCSK9. Thus, the mechanism for the PCSK9-mediated degradation of the LDLR does not appear to involve an interaction between the endosomal sorting machinery and LDLR-specific motifs in the cytoplasmic domain. Moreover, ubiquitination of lysines in the cytoplasmic domain does not appear to play a critical role in the PCSK9-mediated degradation of the LDLR.

Background Familial hypercholesterolemia (FH) patients with the splice site mutation c.313+1, G>A in intron 3 of the low density lipoprotein receptor (LDLR) gene, present with a phenotype similar to that of FH patients in general. However, a mild phenotype would have been expected from the published data showing that the mutation only causes skipping of exon 3. Methods Epstein Barr virus-transformed lymphocytes from eight c.313+1, G>A heterozygotes and two c.313+1, G>A homozygotes were subjected to studies of the LDLR at the mRNA and protein levels. Results Mutation c.313+1, G>A not only causes skipping of exon 3, but also causes inclusion of intron 3. No functional LDLR was produced from the transcript with inclusion of intron 3. The transcript with skipping of exon 3 produced a receptor which had markedly reduced ability to internalize low density lipoprotein. Conclusion The findings that the mutation c.313+1, G>A in the LDLR gene also generates a mutant transcript with inclusion of intron 3, explains why the mutation c.313+1, G>A may result in a severe phenotype.

What the rule of law means and how it constrains the exercise of state power raise issues which have been debated-without resolution-over the ages. Times of emergency bring fresh energy to the discussion, and David Dyzenhaus is one of many who have entered the fray to debate the balance between liberty and national security in the post 9/11 period. It has not been easy for those who place their trust in written constitutions to account for the way textual guarantees are diluted when the state is under threat. Rather than address that dilemma, Dyzenhaus sets his ideas apart by proposing a theory which maximizes the protection of rights in emergency circumstances, without straining the institutional capacities or legitimacy of the judiciary. This theory invokes the pedigree of the common law-and “common law constitutionalism”-and is grounded in the constitutive properties of the rule of law, or principle of legality. Dyzenhaus may not have answered the questions readers will want to ask, but he has opened up the middle ground between the competing supremacies yet more, by drawing common law constitutionalism and its rule-of-law pedigree into constitutional theories of review. More to the point, he has challenged the judiciary to draw on the moral resources of the law to make executive and legislative action as accountable as possible at all times, in emergencies as well as in normal times. Readers can and should engage, at many levels, with the complexity of his thought in this important book.

Problem Gamblers, by Sasso and Kalajdzic, asks whether the province and its gaming venues can be held liable for the losses of those who do not or cannot control their gambling impulses. This article provides a counterpoint to Problem Gamblers. Specifically, it examines the soundness of Sasso and Kalajdzic’s claim that the province and its gaming venues can be held responsible for gambling losses.

The proprotein convertase subtilisin/kexin type 9 (PCSK9) gene is involved in the post-transcriptional regulation of the low-density lipoprotein (LDL) receptors (LDLR). Mutations in the PCSK9 gene have been associated with both hypocholesterolemia and hypercholesterolemia through 'loss-of-function' and 'gain-of-function' mechanisms, respectively. We have studied the effect of the four loss-of-function mutations R46L, G106R, N157K and R237W and the two gain-of-function mutations S127R and D374Y on the autocatalytic activity of PCSK9, as well as on the amount of the cell surface LDLR and internalization of LDL in transiently transfected HepG2 cells. The two groups of mutations did not differ with respect to autocatalytic activity of PCSK9, but they did differ with respect to the amount of cell surface LDLR and internalization of LDL. The four loss-of-function mutations had a 16% increased level of cell surface LDLR and a 35% increased level of internalization of LDL as compared with WT-PCSK9. The two gain-of-function mutations had a 23% decreased level of cell surface LDLR and a 38% decreased level of internalization of LDL as compared with WT-PCSK9. Our studies have also shown that transfer of media from transiently transfected HepG2 cells to untransfected HepG2 cells, reduces the amount of cell surface LDLR and internalization of LDL in the untransfected cells within 20 min of media transfer. Thus, PCSK9 or a factor acted upon by PCSK9, is secreted from the transfected cells and degrades LDLR both in transfected and untransfected cells.

The Charter’s thirtieth anniversary is a checkpoint, an opportunity to take stock of s.2(b)’s journey to date. The focus in this paper is freedom of the press, and the central concern the central concern which emerges is the Supreme Court’s avoidance of s.2(b) and its reluctance to grant newsgathering activities status under the Charter. Insisting that the press can be protected in other ways is unconvincing in the face of the Court’s unwillingness to engage the Charter’s explicit guarantee of a free press. Looking ahead, two obstacles stand in the way of robust protection for this freedom: one is theoretical and calls for a concept that explains the constitutional status of the press; the other is definitional and as such seeks a conceptual answer to the challenge of identifying who the press is at this moment in time. Addressing these obstacles will not only enable but also require the Supreme Court to protect the press and its newsgathering activities under s.2(b). If and when that happens, the status of the press under s.2(b) should be much improved at the Charter’s next checkpoint.

This article on the McLachlin Court and the Charter in 2012 present the annual review of the Supreme Court’s jurisprudence for Osgoode Hall’s Annual Constitutional Cases conference and conference publication (63 Supreme Court L. Rev. (2d); forthcoming, fall 2013). The article provides quantitative data on the 2012 Charter jurisprudence and identifies a downward trend in Charter success rates in the McLachlin Court, from 2010 to the present. The article also offers pointed analysis of two key Charter decisions in this period: R. v. Khawaka and Saskatchewan (Human Rights Commission) v. Whatcott (decided early in 2013). Both raised high stakes questions about the Charter’s fundamental freedoms, and the Court rejected the Charter claims both times, by unanimous decision. The downward trend in Charter success rates, the Court’s unanimity in those key decisions, and the Charter’s mediocre year at the Court inspire a broader reflection, at the end of the article, of the dynamics at work and what they signify. That reflection considers whether the transformative energy of the Charter’s early years has been spent and is being replaced by a conception of the Charter as a middle-ground document. It explores the significance of that shift, for the Charter as an instrument of progress, and for the Court’s leadership in thinking deeply about the Charter’s promises and how they can be kept.

Objective To study whether mutations in the SORT1 gene could be a cause of autosomal dominant hypercholesterolemia and to study the effect of sortilin on the binding and internalization of low density lipoprotein (LDL). Methods 842 unrelated hypercholesterolemic subjects without mutations in genes known to cause autosomal dominant hypercholesterolemia, were screened for mutations in the SORT1 gene by DNA sequencing. Transfections of wild-type or mutant SORT1 plasmids in HeLa T-REx cells and the use of siRNA were used to study the effect of sortilin on the number of cell-surface LDL receptors and on the binding and internalization of LDL. Results A total of 45 mutations in the SORT1 gene were identified of which 15 were missense mutations. Eight of these were selected for in vitro studies, of which none had a major impact on the amount of LDL bound to the cell surface. There was a positive correlation between the amount of sortilin on the cell surface and the amount of LDL bound. The observation that a mutant sortilin which is predominantly found on the cell surface rather than in post-Golgi compartments, bound very high amounts of LDL, indicates that sortilin does not increase the binding of LDL through an intracellular mechanism. Rather, our data indicate that sortilin binds LDL on the cell surface. Conclusion Even though sortilin binds and internalizes LDL by receptor-mediated endocytosis, mutations in the SORT1 gene are unlikely to cause autosomal dominant hypercholesterolemia and may only have a marginal effect on plasma LDL cholesterol levels.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) post-translationally regulates the number of cell-surface low density lipoprotein receptors (LDLR). This is accomplished by the ability of PCSK9 to mediate degradation of the LDLR. The underlying mechanism involves binding of secreted PCSK9 to the epidermal growth factor-like repeat A of the extracellular domain of the LDLR at the cell surface, followed by lysosomal degradation of the internalized LDLR:PCSK9 complex. However, the mechanism by which the normal recycling of the LDLR is disrupted by PCSK9, remains to be determined. In this study we have investigated the role of the cytoplasmic domain of the LDLR for this process. This has been done by studying the ability of a mutant LDLR (K811X-LDLR) which lacks the cytoplasmic domain, to be degraded by PCSK9. We show that this mutant receptor is degraded by PCSK9. Thus, the machinery which directs the LDLR:PCSK9 complex to the lysosomes for degradation, does not interact with the cytoplasmic domain of the LDLR.

The question this article poses is whether the common law must adopt Charter-specific doctrines or remedies when Charter values are at stake. The discussion focuses on the Supreme Court’s defamation decisions, but includes brief remarks about R. v. National Post, which considered whether the Wigmore test for a journalist-source privilege is consistent with the Charter.

Background Inherited low levels of high density lipoprotein (HDL) cholesterol may be due to mutations in the genes encoding the ATP-binding cassette transporter A1 (ABCA1), apolipoprotein (apo) A-I or lecithin:cholesterol acyltransferase (LCAT). Methods The ABCA1, apoA-I and LCAT genes of a 40-year-old male subject with serum HDL cholesterol of 0.06 mmol/l were subjected to DNA sequencing. The proband's family was examined for co-segregation between mutations and levels of HDL cholesterol. Cholesterol efflux in fibroblasts from the proband and a normocholesterolemic subject was compared. The effects of an ABCA1 mutation on cholesterol efflux and membrane localization of ABCA1 were studied in transfected HEK293 and HeLa cells, respectively. Results The proband was a compound heterozygote for ABCA1 mutations R282X (c.844 C>T) and Y1532C (c.4595 A>G). Relatives who were heterozygous for one of these mutations, had about half-normal HDL cholesterol levels. Cholesterol efflux was reduced in fibroblasts from the proband, as was cholesterol efflux from HEK293 cells transfected with an human (h) ABCA1 expression plasmid harboring the Y1532C mutation. Confocal microscopy of HeLa cells transfected with the Y1532C–hABCA1 plasmid revealed that the Y1532C mutation inhibits ABCA1 from reaching the cellular membrane. Conclusion Compound heterozygosity for the nonsense mutation R282X and the missense mutation Y1532C in the ABCA1 gene causes Tangier disease. R282X has a detrimental effect on the function of ABCA1 since a premature stop codon is introduced. Mutation Y1532C disrupts the normal function of ABCA1 as determined by in vitro analyses.