Enzyme-induced Aggregation of Plasmonic Nanoparticles for the Development of Label-free and Ultrasensitive Detection of Bacterial DNA

Sensitive detection of pathogens is critical to ensure the safety of food supplies and to prevent bacterial disease infection and outbreak at the first onset. While conventional techniques such as cell culture, ELISA, PCR, etc. have been used as the predominant detection workhorses, they are however limited by either time-consuming procedure, complicated sample pre-treatment, expensive analysis and operation, or inability to be implemented at point-of-care testing. Here, we present our recently developed assay exploiting enzyme-induced aggregation of plasmonic gold nanoparticles (AuNPs) for label-free and ultrasensitive detection of bacterial DNA. In the experiments, AuNPs are first functionalized with specific, single-stranded RNA probes so that they exhibit high stability in solution even under high electrolytic condition thus exhibiting red color. When bacterial DNA is present in a sample, a DNA-RNA heteroduplex will be formed and subsequently prone to the RNase H cleavage on the RNA probe, allowing the DNA to liberate and hybridize with another RNA strand. This continuously happens until all of the RNA strands are cleaved, leaving the nanoparticles ‘unprotected’. The addition of NaCl will cause the ‘unprotected’ nanoparticles to aggregate, initiating a colour change from red to blue. The reaction is performed in a multi-well plate format, and the distinct colour signal can be discriminated by naked eye or simple optical spectroscopy. As a result, bacterial DNA as low as pM could be unambiguously detected, suggesting that the enzyme-induced aggregation of AuNPs assay is very easy to perform and sensitive, it will significantly benefit to development of fast and ultrasensitive methods that can be used for disease detection and diagnosis.

Progress and Research in Cybersecurity - Supporting a resilient and trustworthy system for the UK

Executive summary
Digital systems have transformed, and will continue to transform, our world. Supportive government policy, a strong research base and a history of industrial success make the UK particularly well-placed to realise the benefits of the emerging digital society. These benefits have already been substantial, but they remain at risk. Protecting the benefits and minimising the risks requires reliable and robust cybersecurity, underpinned by a strong research and translation system.
Trust is essential for growing and maintaining participation in the digital society. Organisations earn trust by acting in a trustworthy manner: building systems that are reliable and secure, treating people, their privacy and their data with respect, and providing credible and comprehensible information to help people understand how secure they are.
Resilience, the ability to function, adapt, grow, learn and transform under stress or in the face of shocks, will help organisations deliver systems that are reliable and secure. Resilient organisations can better protect their customers, provide more useful products and services, and earn people’s trust.
Research and innovation in industry and academia will continue to make important contributions to creating this resilient and trusted digital environment. Research can illuminate how best to build, assess and improve digital systems, integrating insights from different disciplines, sectors and around the globe. It can also generate advances to help cybersecurity keep up with the continued evolution of cyber risks.
Translation of innovative ideas and approaches from research will create a strong supply of reliable, proven solutions to difficult to predict cybersecurity risks. This is best achieved by maximising the diversity and number of innovations that see the light of day as products.
Policy, practice and research will all need to adapt. The recommendations made in this report seek to set up a trustworthy, self-improving and resilient digital environment that can thrive in the face of unanticipated threats, and earn the trust people place in it.
Innovation and research will be particularly important to the UK’s economy as it establishes a new relationship with the EU. Cybersecurity delivers important economic benefits, both by underpinning the digital foundations of UK business and trade and also through innovation that feeds directly into growth. The findings of this report will be relevant regardless of how the UK’s relationship to the EU changes.
Headline recommendations
● Trust: Governments must commit to preserving the robustness of encryption, including end-to-end encryption, and promoting its widespread use. Encryption is a foundational security technology that is needed to build user trust, improve security standards and fully realise the benefits of digital systems.
● Resilience: Government should commission an independent review of the UK’s future cybersecurity needs, focused on the institutional structures needed to support resilient and trustworthy digital systems in the medium and longer term. A self-improving, resilient digital environment will need to be guided and governed by institutions that are transparent, expert and have a clear and widely-understood remit.
● Research: A step change in cybersecurity research and practice should be pursued; it will require a new approach to research, focused on identifying ambitious high-level goals and enabling excellent researchers to pursue those ambitions. This would build on the UK's existing strengths in many aspects of cybersecurity research and ultimately help build a resilient and trusted digital sector based on excellent research and world-class expertise.
● Translation: The UK should promote a free and unencumbered flow of cybersecurity ideas from research to practical use and support approaches that have public benefits beyond their short term financial return. The unanticipated nature of future cyber threats means that a diverse set of cybersecurity ideas and approaches will be needed to build resilience and adaptivity. Many of the most valuable ideas will have broad security benefits for the public, beyond any direct financial returns.

Genetics and Prognostication in Splenic Marginal Zone Lymphoma: Revelations from Deep Sequencing

Purpose: Mounting evidence supports the clinical significance of gene mutations and immunogenetic features in common mature B-cell malignancies.

Experimental Design: We undertook a detailed characterization of the genetic background of splenic marginal zone lymphoma (SMZL), using targeted resequencing and explored potential clinical implications in a multinational cohort of 175 patients with SMZL.

Results: We identified recurrent mutations in TP53 (16%), KLF2 (12%), NOTCH2 (10%), TNFAIP3 (7%), MLL2 (11%), MYD88 (7%), and ARID1A (6%), all genes known to be targeted by somatic mutation in SMZL. KLF2 mutations were early, clonal events, enriched in patients with del(7q) and IGHV1-2*04 B-cell receptor immunoglobulins, and were associated with a short median time to first treatment (0.12 vs. 1.11 years; P = 0.01). In multivariate analysis, mutations in NOTCH2 [HR, 2.12; 95% confidence interval (CI), 1.02–4.4; P = 0.044] and 100% germline IGHV gene identity (HR, 2.19; 95% CI, 1.05–4.55; P = 0.036) were independent markers of short time to first treatment, whereas TP53 mutations were an independent marker of short overall survival (HR, 2.36; 95 % CI, 1.08–5.2; P = 0.03).

Conclusions: We identify key associations between gene mutations and clinical outcome, demonstrating for the first time that NOTCH2 and TP53 gene mutations are independent markers of reduced treatment-free and overall survival, respectively.

Effect of intravenous haloperidol on the duration of delirium and coma in critically ill patients (Hope-ICU):A randomised, double-blind, placebo-controlled trial

Background: Delirium is frequently diagnosed in critically ill patients and is associated with poor clinical outcomes. Haloperidol is the most commonly used drug for delirium despite little evidence of its effectiveness. The aim of this study was to establish whether early treatment with haloperidol would decrease the time that survivors of critical illness spent in delirium or coma. Methods: We did this double-blind, placebo-controlled randomised trial in a general adult intensive care unit (ICU). Critically ill patients (≥18 years) needing mechanical ventilation within 72 h of admission were enrolled. Patients were randomised (by an independent nurse, in 1:1 ratio, with permuted block size of four and six, using a centralised, secure web-based randomisation service) to receive haloperidol 2·5 mg or 0·9% saline placebo intravenously every 8 h, irrespective of coma or delirium status. Study drug was discontinued on ICU discharge, once delirium-free and coma-free for 2 consecutive days, or after a maximum of 14 days of treatment, whichever came first. Delirium was assessed using the confusion assessment method for the ICU (CAM-ICU). The primary outcome was delirium-free and coma-free days, defined as the number of days in the first 14 days after randomisation during which the patient was alive without delirium and not in coma from any cause. Patients who died within the 14 day study period were recorded as having 0 days free of delirium and coma. ICU clinical and research staff and patients were masked to treatment throughout the study. Analyses were by intention to treat. This trial is registered with the International Standard Randomised Controlled Trial Registry, number ISRCTN83567338. Findings: 142 patients were randomised, 141 were included in the final analysis (71 haloperidol, 70 placebo). Patients in the haloperidol group spent about the same number of days alive, without delirium, and without coma as did patients in the placebo group (median 5 days [IQR 0-10] vs 6 days [0-11] days; p=0·53). The most common adverse events were oversedation (11 patients in the haloperidol group vs six in the placebo group) and QTc prolongation (seven patients in the haloperidol group vs six in the placebo group). No patient had a serious adverse event related to the study drug. Interpretation: These results do not support the hypothesis that haloperidol modifies duration of delirium in critically ill patients. Although haloperidol can be used safely in this population of patients, pending the results of trials in progress, the use of intravenous haloperidol should be reserved for short-term management of acute agitation. Funding: National Institute for Health Research. © 2013 Elsevier Ltd.