Novel Kraft Softwood Lignin-Derived Carbon Quantum Dots: Synthesis, Characterization, and In Vitro Cytocompatibility.

Carbon quantum dots (CQDs) have been investigated for biomedical applications in medical imaging due to their fluorescent properties, overall long-term stability, and excellent cytocompatibility and biocompatibility. Lignin is an organic polymer in the tissues of woody plants. It is also considered a byproduct of the wood and pulp industries. Hence, it presents as a renewable source of carbon nanoparticles. In this study, we report the synthesis and material and biological characterization of two colloidal suspensions of CQDs in water derived from lignin-based carbon. One was the native form of CQDs derived from lignin carbon, and the second was doped with nitrogen to evaluate material differences. Material characterization was carried out using various commonly used techniques, including Fourier transform infrared spectroscopy (FTIR), emission and absorbance spectra, zeta potential, transmission electron microscopy (TEM), and scanning electron microscopy (SEM). Thin films of CQDs were formed on glass and silicon substrates to assess the in vitro cytocompatibility with human mesenchymal stem cells (hMSCs). Observations suggest that the two forms of CQDs promote cell attachment within 24 h and sustain it for at least 7 days. The overall structure and shape of cells suggest a lack of any adverse or toxic effects of CQDs. The data lay down the novel foundation to support the use of lignin-derived CQDs in tissue engineering applications.

Criminal Code reform of HIV non-disclosure is urgently needed: Social science perspectives on the harms of HIV criminalization in Canada.

The criminalization of HIV non-disclosure represents a significant issue of concern among people living with HIV, those working across the HIV sector, public health practitioners, and health and human rights advocates around the world. Recently, the government of Canada began a review of the criminal law regarding HIV non-disclosure and invited feedback from the public about potential reforms to the Criminal Code. In light of this public consultation, this commentary examines social science research from Canadian scholars that documents the intersecting damaging effects of HIV criminalization. Canadian social scientists and other researchers have shown that HIV criminalization is applied in uneven and discriminatory ways, impedes HIV prevention efforts, perpetuates HIV stigma, and has a damaging impact on the daily lives of people living with HIV. We argue that there is an urgent need for reforms that will significantly restrict how the criminal law is applied to HIV non-disclosure.

RéSUMé: La criminalisation de la non-divulgation du VIH est une question très préoccupante pour les personnes vivant avec le VIH, celles qui travaillent dans le secteur du VIH, les praticiens et praticiennes de la santé publique et les porte-parole de la santé et des droits de la personne du monde entier. Récemment, le gouvernement du Canada a amorcé un examen du droit criminel portant sur la non-divulgation du VIH et a invité le public à commenter d'éventuelles réformes du Code criminel. À la lumière de cette consultation publique, notre commentaire porte sur les études en sciences sociales menées au Canada qui font état des effets croisés préjudiciables de la criminalisation du VIH. Des spécialistes des sciences sociales et d'autres chercheuses et chercheurs canadiens ont montré que la criminalisation du VIH est appliquée de façon inégale et discriminatoire, qu'elle nuit aux efforts de prévention du VIH, qu'elle perpétue la stigmatisation liée au VIH et qu'elle a des effets dommageables sur la vie quotidienne des personnes vivant avec le VIH. Nous soutenons qu'il existe un besoin urgent de réformes pour restreindre de façon appréciable l'application du droit criminel à la non-divulgation du VIH.

When biographical disruption meets HIV exceptionalism: Reshaping illness identities in the shadow of criminalization.

Drawing on interviews with civil society actors in the AIDS Service Organization (ASO) sector in Canada, this article explores how these actors contribute to shaping the illness identities of people living with HIV/AIDS in the shadow of efforts to criminalize exposure to HIV. While the biographically disruptive qualities associated with an HIV diagnosis have been addressed in the medical sociology literature, we turn our attention to the key role played by ASOs as interlocutors in this process. Paying specific attention to the intersection of processes of medicalization and criminalization, we ask how they are re-stigmatizing a condition that has shifted in the public consciousness from its earlier association with deviance and moral culpability. One important implication of our findings concerns the need to take greater account of how the illness identity and experience can be shaped by a 'biography of telling', of a renewed pressure to disclose intimate details of one's health status as a way to perform responsible practices of citizenship.

Governing COVID-19 without government in Brazil: Ignorance, neoliberal authoritarianism, and the collapse of public health leadership.

Brazil's governance of the COVID-19 pandemic has been described as nothing short of tragic by several commentators. President Jair Bolsonaro's dangerous brew of neoliberal authoritarianism, science denialism and ableism has plunged this country into catastrophe. In this article we argue that this form (or lack) of public health governance can best be described as governance without (central) government. We begin with an overview of public health governance in the country before introducing the main theoretical concepts that guide our analysis, namely the notions of 'government by exception' and 'strategic ignorance'. Finally, we sketch the main features of this emerging form of (non)governance of COVID-19. We highlight the new forms of solidarity and mutual aid that have emerged in favelas and Indigenous communities, which have stepped in to fill the void left by a limited federal presence. The article concludes by reflecting on what this collapse of public health reveals about the limitations of democratic governance in the age of Bolsonaro.

"We write as little as we have to": charting practices and documenting disclosure in response to HIV criminalization in Canada.

The criminalization of HIV nondisclosure is reshaping the landscape of support and care for people living with HIV/AIDS (PLWHA). Focusing on Canada, this article examines how criminalization is reshaping the relationships between frontline AIDS Service Organization (ASO) workers and their HIV-positive service users. Using data gleaned through semi-structured interviews with N = 62 frontline ASO staff members across Canada that were coded using thematic analysis, we describe how ASO workers are rethinking and altering their notetaking practices in light of fears about criminalization and how their charts may be used by legal actors. Specifically, we identified three main themes. First, awareness that their notes can be subpoenaed and used against their clients in criminal proceedings is leading ASO staff to keep less detailed records of their conversations with PLWHA, although there were variations by the type of position held. Participants with professional obligations (doctors, nurses, social workers) reported that they continue to keep more detailed charting records than other frontline ASO staff (peer counselors, educators, etc.). Second, participants acknowledged that criminalization threatens the trust dynamic in the therapeutic relationship and that by taking less detailed notes they risk the quality of care provided. Third, we found that evolving knowledge about risk and responsibility in light of criminalization is impacting the type of counsel ASO workers offer to PLWHA regarding how to document disclosure. The article concludes by reflecting on the challenges associated with navigating the "medico-legal borderland", as ASO staff balance their commitments to serving PLWHA with the realities created by the harsh climate of criminalization. Greater political attention should be paid to the risks that criminalization poses to frontline HIV support work and efforts to support PLWHA, who already face significant stigma, without the additional stigma of the criminal label.

High-Throughput Screening Strategy Identifies Allosteric, Covalent Human D-Amino Acid Oxidase Inhibitor.

Genome-wide association studies have linked polymorphisms in the gene G72 to schizophrenia risk in several human populations. Although controversial, biochemical experiments have suggested that the mechanistic link of G72 to schizophrenia is due to the G72 protein product, pLG72, exerting a regulatory effect on human D-amino acid oxidase (hDAAO) activity. In an effort to identify hDAAO inhibitors of novel mechanism of action, we designed a pLG72-directed hDAAO activity assay suitable for high-throughput screening (HTS). During assay development, we confirmed that pLG72 was an inhibitor of hDAAO. Thus, our assay employed an IC20 pLG72 concentration that was high enough to allow dynamic pLG72-hDAAO complexes to form but with sufficient remaining hDAAO activity to measure during an HTS. After conducting an approximately 150,000-compound HTS, we further characterized a class of compound hits that were less potent hDAAO inhibitors when pLG72 was present. Focusing primarily on compound 2: [2-(2,5-dimethylphenyl)-6-fluorobenzo[d]isothiazol-3(2H)-on], we demonstrated that these compounds inhibited hDAAO via an allosteric, covalent mechanism. Although there is significant interest in the therapeutic potential of compound 2: and its analogues, their sensitivity to reducing agents and their capacity to bind cysteines covalently would need to be addressed during therapeutic drug development.

Globalization and the health of Canadians: 'Having a job is the most important thing'.

BACKGROUND: Globalization describes processes of greater integration of the world economy through increased flows of goods, services, capital and people. Globalization has undergone significant transformation since the 1970s, entrenching neoliberal economics as the dominant model of global market integration. Although this transformation has generated some health gains, since the 1990s it has also increased health disparities. METHODS: As part of a larger project examining how contemporary globalization was affecting the health of Canadians, we undertook semi-structured interviews with 147 families living in low-income neighbourhoods in Canada's three largest cities (Montreal, Toronto and Vancouver). Many of the families were recent immigrants, which was another focus of the study. Drawing on research syntheses undertaken by the Globalization Knowledge Network of the World Health Organization's Commission on Social Determinants of Health, we examined respondents' experiences of three globalization-related pathways known to influence health: labour markets (and the rise of precarious employment), housing markets (speculative investments and affordability) and social protection measures (changes in scope and redistributive aspects of social spending and taxation). Interviews took place between April 2009 and November 2011. RESULTS: Families experienced an erosion of labour markets (employment) attributed to outsourcing, discrimination in employment experienced by new immigrants, increased precarious employment, and high levels of stress and poor mental health; costly and poor quality housing, especially for new immigrants; and, despite evidence of declining social protection spending, appreciation for state-provided benefits, notably for new immigrants arriving as refugees. Job insecurity was the greatest worry for respondents and their families. Questions concerning the impact of these experiences on health and living standards produced mixed results, with a majority expressing greater difficulty 'making ends meet,' some experiencing deterioration in health and yet many also reporting improved living standards. We speculate on reasons for these counter-intuitive results. CONCLUSIONS: Current trends in the three globalization-related pathways in Canada are likely to worsen the health of families similar to those who participated in our study.

A selective small molecule NOP (ORL-1 receptor) partial agonist for the treatment of anxiety.

Small molecule (1) has been identified as a selective partial agonist of Opioid Receptor Like-1 (ORL-1) with potential utility for the treatment of anxiety and other disorders. Nociceptin (orphanin FQ) is an endogenous peptide ligand that binds to ORL-1, however it does not bind the classical δ, µ and κ opioid receptors with high affinity. The synthesis of 1 involved using a molecular diversity approach, to rapidly advance a library of compounds for biological testing. A lead selective potent partial agonist (35-fold ORL-1/Mu) progressed to ORL-1 (NOP or OP4) proof of concept testing in advanced studies. The synthetic approach and biological data for the related chemical series will be presented.

Novel human D-amino acid oxidase inhibitors stabilize an active-site lid-open conformation.

The NMDAR (N-methyl-D-aspartate receptor) is a central regulator of synaptic plasticity and learning and memory. hDAAO (human D-amino acid oxidase) indirectly reduces NMDAR activity by degrading the NMDAR co-agonist D-serine. Since NMDAR hypofunction is thought to be a foundational defect in schizophrenia, hDAAO inhibitors have potential as treatments for schizophrenia and other nervous system disorders. Here, we sought to identify novel chemicals that inhibit hDAAO activity. We used computational tools to design a focused, purchasable library of compounds. After screening this library for hDAAO inhibition, we identified the structurally novel compound, 'compound 2' [3-(7-hydroxy-2-oxo-4-phenyl-2H-chromen-6-yl)propanoic acid], which displayed low nM hDAAO inhibitory potency (Ki=7 nM). Although the library was expected to enrich for compounds that were competitive for both D-serine and FAD, compound 2 actually was FAD uncompetitive, much like canonical hDAAO inhibitors such as benzoic acid. Compound 2 and an analog were independently co-crystalized with hDAAO. These compounds stabilized a novel conformation of hDAAO in which the active-site lid was in an open position. These results confirm previous hypotheses regarding active-site lid flexibility of mammalian D-amino acid oxidases and could assist in the design of the next generation of hDAAO inhibitors.

Structural, kinetic, and pharmacodynamic mechanisms of D-amino acid oxidase inhibition by small molecules.

We characterized the mechanism and pharmacodynamics of five structurally distinct inhibitors of d-amino acid oxidase. All inhibitors bound the oxidized form of human enzyme with affinity slightly higher than that of benzoate (Kd ≈ 2-4 µM). Stopped-flow experiments showed that pyrrole-based inhibitors possessed high affinity (Kd ≈ 100-200 nM) and slow release kinetics (k < 0.01 s(-1)) in the presence of substrate, while inhibitors with pendent aromatic groups altered conformations of the active site lid, as evidenced by X-ray crystallography, and showed slower kinetics of association. Rigid bioisosteres of benzoic acid induced a closed-lid conformation, had slower release in the presence of substrate, and were more potent than benzoate. Steady-state d-serine concentrations were described in a PK/PD model, and competition for d-serine sites on NMDA receptors was demonstrated in vivo. DAAO inhibition increased the spatiotemporal influence of glial-derived d-serine, suggesting localized effects on neuronal circuits where DAAO can exert a neuromodulatory role.

Design, synthesis, and structure-activity relationships of novel bicyclic azole-amines as negative allosteric modulators of metabotropic glutamate receptor 5.

A novel series of diaryl bicyclic azole-amines that are potent selective negative modulators of metabotropic glutamate receptor 5 (mGluR5) were identified through rational design. An initial hit compound 5a of modest potency (IC(50) = 1.2 µM) was synthesized. Evaluation of structure-activity relationships (SAR) on the left-hand side of the molecule revealed a preference for a 2-substituted pyridine group linked directly to the central heterocycle. Variation of the central azolo-amine portion of the molecule revealed a preference for the [4,5-c]-oxazoloazepine scaffold, while right-hand side variants showed a preference for ortho- and meta-substituted benzene rings linked directly to the tertiary amine of the saturated heterocycle. These iterations led to the synthesis of 29b, a potent (IC(50) = 16 nM) and selective negative modulator that showed good brain penetrance, high receptor occupancy, and a duration of action greater than 1 h in rat when administered intraperitoneally. Formal PK studies in rat and Rhesus monkey revealed a short half-life that was attributable to high first-pass clearance.

The unintended consequences of being friendly: a case study.

Baby-friendly certification recognizes hospitals that promote early physical bonding between mother and infant, immediately after birth. Most births can accomplish physical bonding without increased risk to mother or infant. When mother or infant have complications and each have intravenous (i.v.) lines and are receiving medications, the physical bonding post-birth may also inadvertently put the patients at risk. A baby-friendly community hospital in New England found that early bonding put an infant at higher risk for medication error when the two i.v. lines were not properly identified and the infant received a medication intended for the mother. The growing body of literature on i.v. medication safety does not address this particular type of error, and this was an error that technology would not have prevented. The "5 rights" of medication safety are not as effective as physical separation of the two individuals during medication administration. A brief separation does not diminish bonding, and the practice has prevented subsequent errors.

Identifying nonselective hits from a homogeneous calcium assay screen.

The authors used a homogeneous calcium dye kit with a cell line transfected using a recombinant protein construct to screen a 50,000 compound library for G-protein coupled receptor (GPCR) agonists. Only 1 of the 365 primary hits activated Gq-coupled GPCRs, as shown using IP-ONE HTRF. Furthermore, an agonist screen against the entire compound library and same heterologous cell line using AequoScreen technology generated no false positives and identified the same positive hit. Next, a multiplex assay composed of both Fluo-3 and Fura-2-loaded cells identified 1 false positive and the same true-positive hit out of the 365 primary hits. Finally, rescreening the 365 primary hits against the parental cell line loaded using the homogeneous calcium dye kit confirmed the specificity of the same true-positive hit only. In summary, the results suggest that AequoScreen technology, IP-ONE HTRF, and multiplex assays are unique, orthogonal technologies to identify nonspecific hits.

Metastin (KiSS-1) mimetics identified from peptide structure-activity relationship-derived pharmacophores and directed small molecule database screening.

Metastin, also known as KiSS-1, the cognate ligand for the metastin receptor GPR54, is a peptide known to dramatically reduce metastasis in experimental models. Despite this, there is no reported structure for metastin nor any small molecule modulators of metastin function that could be used either clinically or experimentally. Here we report the NMR solution structure of a 13-residue metastin peptide in a membrane-like environment (SDS micelles) and find it to have a relatively stable helix conformation from residues 7 to 13. In assays for metastin receptor binding and calcium flux with receptor-transfected HEK-293 cells, we demonstrate through alanine scanning and amino acid substitutions that the peptide C-terminus shows helix periodicity in an NMR structural model and that Phe9, Arg12, and Phe13 are crucial to the activity of the peptide. These three residues lie on one face of the helix and define a pharmacophore site for metastin. We used these pharmacophore features in small molecule database searches to identify hits with submicromolar affinity for the metastin receptor. We also show here that molecules mimicking key elements of this pharmacophore site bind to the metastin receptor and act as full agonists, albeit with reduced potency compared to that of metastin itself. Together this structure-activity approach may yield pharmacologically useful compounds relevant in defining and modulating metastin receptor function.

3-(4-Piperidinyl)indoles and 3-(4-piperidinyl)pyrrolo-[2,3-b]pyridines as ligands for the ORL-1 receptor.

A novel series of indoles and 1H-pyrrolo[2,3-b]pyridines having a piperidine ring at the 3-position were synthesized and found to bind with high affinity to the ORL-1 receptor. Structure-activity relationships at the piperidine nitrogen were investigated in each series. Substitution on the phenyl ring and nitrogen atom of the indole and 1H-pyrrolo[2,3-b]pyridine cores generated several selective high-affinity ligands that were agonists of the ORL-1 receptor.

Medication safety--reliability of preference cards.

A CLINICAL ANALYSIS of surgeons' preference cards was initiated in one hospital as part of a comprehensive analysis to reduce medication-error risks by standardizing and simplifying the intraoperative medication-use process specific to the sterile field. THE PREFERENCE CARD ANALYSIS involved two subanalyses: a review of the information as it appeared on the cards and a failure mode and effects analysis of the process involved in using and maintaining the cards. THE ANALYSIS FOUND that the preference card system in use at this hospital is outdated. Variations and inconsistencies within the preference card system indicate that the use of preference cards as guides for medication selection for surgical procedures presents an opportunity for medication errors to occur.

Preparation of 3-spirocyclic indolin-2-ones as ligands for the ORL-1 receptor.

A novel series of indolin-2-ones having a spirocyclic piperidine ring at the 3-position was synthesized and found to bind with high affinity to the ORL-1 receptor. Structure-activity relationships at the piperidine nitrogen were investigated. Substitution on the phenyl ring and nitrogen atom of the indolin-2-one core generated several selective high-affinity ligands that were antagonists of the ORL-1 receptor.