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Wastewater-based genomic surveillance of the SARS-CoV-2 virus shows promise to complement genomic epidemiology efforts. Multiplex tiled PCR is a desirable approach for targeted genome sequencing of SARS-CoV-2 in wastewater due to its low cost and rapid turnaround time. However, it is not clear how different multiplex tiled PCR primer schemes or wastewater sample matrices impact the resulting SARS-CoV-2 genome coverage. The objective of this work was to assess the performance of three different multiplex primer schemes, consisting of 150bp, 400bp, and 1200bp amplicons, as well as two wastewater sample matrices, influent wastewater and primary sludge, for targeted genome sequencing of SARS-CoV-2. Wastewater samples were collected weekly from five municipal wastewater treatment plants (WWTPs) in the Metro Vancouver region of British Columbia, Canada during a period of increased COVID-19 case counts from February to April, 2021. RNA extracted from clarified influent wastewater provided significantly higher genome coverage (breadth and median depth) than primary sludge samples across all primer schemes. Shorter amplicons appeared more resilient to sample RNA degradation, but were hindered by greater primer pool complexity in the 150bp scheme. The identified optimal primer scheme (400bp) and sample matrix (influent) was capable of detecting the emergence of mutations associated with genomic variants of concern, of which the daily wastewater load significantly correlated with clinical case counts. Taken together, these results provide guidance on best practices for implementing wastewater-based genomic surveillance, and demonstrate its ability to inform epidemiology efforts by detecting genomic variants of concern circulating within a geographic region.ImportanceMonitoring the genomic characteristics of the SARS-CoV-2 virus circulating in a population can shed important insights into epidemiological aspects of the COVID-19 outbreak. Sequencing every clinical patient sample in a highly populous area is a difficult feat, and thus sequencing SARS-CoV-2 RNA in municipal wastewater offers great promise to augment genomic surveillance by characterizing a pooled population sample matrix, particularly during an escalating outbreak. Here, we assess different approaches and sample matrices for rapid targeted genome sequencing of SARS-CoV-2 in municipal wastewater. We demonstrate that the optimal approach is capable of detecting the emergence of SARS-CoV-2 genomic variants of concern, with strong correlations to clinical case data in the province of British Columbia. These results provide guidance on best practices on, as well as further support for, the application of wastewater genomic surveillance as a tool to augment current genomic epidemiology efforts.

In this work, we aim to access the performance of three different multiplex primer schemes, i.e. Swift amplicon SARS-CoV-2 panel (150bp amplicons), ARTIC V3 panel (400bp amplicons), and SARS-CoV-2 midnight panel (1200bp amplicons), for metatranscriptomic sequencing of SARS-CoV-2 for influent wastewater and primary sludge. This protocol is adapted from the Swift amplicon™ SARS-COV-2 protocol (150bp amplicon), ARTIC V3 protocol (400bp amplicon), and "midnight" protocol (1200bp amplicon). Sequencing libraries are prepared with 1) Oxford Nanopore Ligation Sequencing Kit (SQK-LSK109) with Native Barcoding kit (EXP-NEB104 and EXP-NEB114), or 2) NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® with NEBNext® Multiplex Oligos for Illumina®. Links to the protocols are: Swift amplicon protocol (150bp): https://swiftbiosci.com/swift-amplicon-sars-cov-2-panel/ ARTIC V3 protocol (400bp): https://www.protocols.io/view/ncov-2019-sequencing-protocol-v3-locost-bh42j8ye?version_warning=no "midnight" protocol V4 (1200bp): dx.doi.org/10.17504/protocols.io.bh7hj9j6

Despite recent advances in diagnosis and treatment, ovarian cancer (OC) is the most lethal gynecological malignancy and improving the efficacy of chemotherapy is of great interest. This study increases our understanding of how dietary intervention with docosahexaenoic acid (DHA)-a supplement proven safe for human consumption – enhances the anti-cancer effects of conventional chemotherapy. Our results demonstrated synergistic cell killing by DHA and carboplatin in OC cell lines. Furthermore, DHA supplementation alone and in combination with carboplatin significantly reduced OC growth in a high-grade serous OC patient-derived xenograft mouse model. Carboplatin administered intraperitoneally significantly reduced tumor growth in DHA-fed mice compared to mice on the control diet. Intravenous carboplatin administration in combination with DHA reduced tumor growth similarly to carboplatin or DHA monotherapies. The DHA-induced reduction in tumor growth in this model was associated with increased tumor necrosis and improved survival. As such, our findings provide a strong rationale to move to clinical trials that will determine whether DHA supplementation enhances the efficacy and tolerance of cytotoxic chemotherapy in patients with OC.

Clinical bottom line An 8-week-old infant is receiving inhaled nitric oxide (iNO) for secondary pulmonary hypertension (PHT) in paediatric intensive care unit (PICU). The child is postcongenital diaphragmatic hernia repair and is proving difficult to wean from iNO. Given the problem with rebound PHT, you wonder if treatment with enteral sildenafil will aid discontinuation of iNO. In the child with pulmonary hypertension (population), does treatment with enteral sildenafil (intervention) compared with a slow wean from iNO alone (comparison) prevent rebound PHT and allow for discontinuation of iNO (outcome)? We performed an online search using PubMed and Medline (1946 to present) in September 2019. The key terms were (pulmonary hypertension) AND (rebound) AND (sildenafil) AND (nitric oxide) AND (child). We limited the search to include articles written in English. We identified three relevant articles, a randomised controlled trial, a prospective and a retrospective …

Rat infestations are common, particularly in impoverished, inner-city neighborhoods. However, there has been little research into the nature and consequences of rat exposure in these neighborhoods, particularly in Canada. In this study, we sought to characterize exposure to rats and rat-associated Leptospira interrogans and Bartonella tribocorum, as well as risk factors associated with exposure, in residents (n = 202) of the Downtown Eastside (DTES) neighborhood of Vancouver, Canada. There was no evidence of exposure to rat-associated L. interrogans but 6/202 (3.0%) of participants were exposed to B. tribocorum, which is known to be circulating among DTES rats. We also found that frequent and close rat exposure was common among DTES residents, and that this exposure was particularly associated with injection drug use and outdoor income-generating activities (e.g., drug dealing). These risk factors may be good targets for interventions geared toward effectively reducing rat exposure.

Aims Airway management in the critically ill child can carry significant risk. We present the development and implementation of Northern Ireland’s first ever Paediatric Emergency airway course. Previously, any training that did exist was on an opportunistic basis. This course aimed to improve patient safety by enhancing the knowledge, skills and behaviours of a multidisciplinary audience with regards to airway management of the critically ill child. Methods A faculty of Paediatric Intensivists, Paediatric Anaesthetists, General Paediatricians and Critical Care Nurses was assembled to develop and deliver the course. The course consisted of three main elements: Presentations outlining the theory of airway management in the critically ill child. Hand–on skill stations with high fidelity paediatric airway mannequins to allow development of technical skills. High fidelity simulation scenarios aiming to develop non technical skills and integrate the knowledge, skills and behaviours addressed in earlier sessions. Results To date the course has run 5 times reaching a total of 128 participants. Participants have included medics and nurses from a range of backgrounds (Paediatric Intensive Care, General Paediatrics, Paediatric Emergency Medicine, Neonatology and Anesthetics) in addition to paramedics. Candidates provide detailed written feedback following each course via a 5 point Likert Scale (5 strongly agree, 1 strongly disagree). Feedback has been excellent with average scores as follows: The course will change my future practice 4.7/5 (94%) The course will make my practice safer 4.9/5 (98%) The course has made me more confident 4.7/5 (94%) The course continues to evolve. It is now run tri-annually and has gained both Difficult Airway Society and RCPCH accreditation. A free online course manual and iBook has been developed to complement the course. Conclusions We have successfully introduced and embedded a multidisciplinary Paediatric Emergency Airway course, into Northern Ireland with excellent uptake and feedback.

In the aftermath of the global financial crisis (‘GFC’), policy-makers and regulators have been engaged in Herculean efforts to restore their tarnished reputations. These efforts have involved jettisoning much of their flawed ideological baggage of old, rooted in a belief about the self-correcting nature of markets and the ability of financial institutions to police themselves, and, in turn, building new, more muscular regulatory and supervisory regimes. This ongoing process is explored through the lens of a high-profile facet of shadow banking, namely the regulation of sale and repurchase agreements (so-called ‘repos’). Here, regulators have set out to address a key vulnerability associated with the maturity mismatch between the short-term liabilities of the repo borrower and its longer-term assets, which has the potential to trigger a so-called ‘repo run’. Through a review of a series of highly problematic reform options, the discussion indicates that there is a danger that regulators will repeat the mistakes of the past by substituting their misplaced faith, pre-GFC, in the self-correcting power of markets, with a misguided conviction, post-crisis, in their capacities to build regulatory regimes which can contain the risks associated with modern-day financial markets. It is suggested that such over-confidence is liable to lull us into a false sense of security about how robust these regimes really are, and to cause regulators to shirk some of the more difficult questions which remain to be addressed.

Using Bismuth Germanate (BGO) scintillators coupled to general purpose analog Silicon Photomultipliers (SiPM) we have developed methods to improve the coincidence timing resolution of PET detectors with BGO in order to reach a timing resolution that can be used for time of flight (TOF) PET detection. TOF measurements have already been acquired using LYSO crystals, but the low cost of BGO along with its optical properties and favorable energy resolution make BGO an attractive option to reduce the overall cost of PET scanners without sacrificing image quality. Using an analog SiPM with a pre-amplifier at a higher voltage improves energy and timing resolution for BGO coincidence detection, but also introduces a much higher baseline noise level, which can obscure the start of the waveform. We have found this noise level can be significantly reduced by cooling the detectors to values below -30°C, allowing us to perform leading edge discrimination (LED) timing at much lower voltages. On average cooling the detectors to this point improves their timing resolution by around 300ps, and as much as 600ps. Using this setup our values are approaching the values necessary for time of flight PET detection. In this paper we will discuss the experimental setup in further depth and share promising initial timing and energy resolution results from our findings.

Univentricular heart disease accounts for ~1.25% of all congenital heart disease. Such cases remain among the most challenging to manage, typically requiring a three-staged palliation. The first stage involves placement of a systemic to pulmonary shunt. While a variety of shunt types, including ductal stenting, can be used to manage univentricular conditions, the archetype remains the Blalock-Taussig (BT) shunt. While waiting future palliative intervention at home, intercurrent illness may necessitate presentation to a district general hospital where subspecialist advice and assessment is remote. This review aims to present the general paediatrician with a straightforward BT shunt physiology overview highlighting unique complications which may complicate intercurrent illness.

This chapter sets out to explore the role of hedge funds in the context of evolving market developments in credit intermediation, in particular, with regard to sale and repurchase transactions (so-called “repos”)—an important and increasingly high profile facet of the shadow banking universe. More pertinently, it seeks to critically analyze the ways in which hedge funds can trigger and, in turn, help to transmit systemic risks in the context of repo transactions. In doing so, the chapter aims to challenge long-maintained claims—both from within the hedge fund sector itself, and amongst certain sections of the academic community—which downplay, or even dismiss, concerns relating to the capacity of hedge funds to cause significant disruption to the wider financial system. The validity or otherwise of these claims has a bearing on the ongoing debate with regard to the need for additional regulatory oversight of hedge fund activities, both in the context of repo transactions and, indeed, more generally.

Pulse oximetry is a first-line monitoring tool, used in neonatal medicine routinely as a part of continuous monitoring during intensive care. It is also used to guide response to resuscitation and as a screening tool for congenital heart disease. Despite its widespread use, many healthcare providers are unaware of the underlying principles and limitations of pulse oximetry in neonates. In this article, we will discuss the physiological and technological principles behind the use of saturation monitoring and its use in neonatal practice.