Deliberate Release Research Articles

The Genetically Modified Organism Medicinal Framework in Europe, United States, and Japan: Underlying Scientific Principles and Considerations Toward the Development of Gene Therapy and Genetically Modified Cell-Based Products.

In vivo viral gene therapy and somatic cell therapy products (whether autologous, allogeneic, or xenogeneic) that have been subjected to an ex vivo gene transfer procedure will normally be classified as genetically modified organisms (GMOs) in Europe, not just the gene transfer vectors used in their construction. These products are, therefore, expected to fulfill certain environmental requirements with regard to the biosafety aspects of their clinical use (which may be subject to review by government departments responsible for environmental affairs). In the European Union (EU), clinical trials using GMOs generally require three levels of review (in addition to local review processes), which are often performed by separate national agencies. In this study, the principles under which certain EU member states control use of the GMOs in clinical trials under the definitions of either "contained use" or "deliberate release" will be discussed and evaluated from a scientific and a regulatory perspective, with comparisons with non-EU expectations as described by the U.S. Food and Drug Administration and the Japanese living modified organisms (LMOs) regulations. For the latter, an understanding of the criteria under which LMOs exemptions apply, notably with respect to the nature of the viral construct used, the manufacturing process, and demonstration that there is no detectable residual replication-competent virus in the final gene-modified cells, is of paramount importance. Building on the existing European, U.S., and Japanese experience with GMOs/LMOs within the context of experimental gene and cell therapies, a through reflection on, and harmonization of, the current global GMO framework is needed to avoid unnecessary delays in clinical development and to ensure a smooth and a rapid access by patients to innovative life-saving therapies.

Themes and correlations of participant experience and evaluation of an interactive bioterrorism release exercise – a mixed methods study

Bioterrorism and pandemics pose great risk to the health and safety of our modern world. Pandemic scenario exercises commonly use diseases that are most likely to cause moderate harm in an epidemic scenario such as influenza. Despite the generalisable nature of most pandemic responses, exercises often fail to take account of the broader impacts of a pandemic scenario. In August 2018, The Exercise Mataika pandemic workshop was conducted by the NHMRC Centre for Research Excellence Integrated Systems for Epidemic Response at the University of New South Wales, Australia. By utilising a high risk, worst case scenario – the deliberate release of Smallpox in Fiji and a much larger Asian country – impacts not often considered in pandemic planning, such as the resiliency of the health system, absenteeism, social cohesion, and broader impacts on society were considered, and compared, across geographic and social groupings. This study aimed to collect and analyse participant perceptions and evaluation of Exercise Mataika. A mixed methods study collecting participants ratings of experience, value and utility aspects of the scenario coupled with a thematic analysis of qualitative responses was conducted. Quantitative ratings for the activity were overwhelmingly positive, with respondents highlighting that the activity was useful, different in format, identified issues not often explored in pandemic exercises, and was a valuable educational and networking opportunity. Qualitative analysis and combined mixed-methods analysis revealed more nuanced findings. While respondents remained positive about the exercise format, subgroups highlighted potential missed opportunities and areas within the scenario where greater focus could have been directed. Overall the findings highlighted the value of including a wide range of exercise attendees both across sectors and nationalities, and addressing a far broader set of considerations across multiple domains. These findings will guide future development of pandemic response exercises and education.

A novel governance framework for GMO: A tiered, more flexible regulation for GMOs would help to stimulate innovation and public debate.

EMBO Reports (2019) e47812 New breeding techniques, in particular gene editing technologies such as CRISPR, have great potential to transform agri‐ and aquaculture. However, realising this potential depends in large part on the regulation of genetically engineered products. Given the rapid technological progress in molecular biology, and the widely diverging interests of stakeholders, there is an urgent need for novel approaches regarding the governance of genetically engineered crops and animals along with a restart of the public dialogue. Importantly, regulation should stimulate technological development while simultaneously maintaining governmental oversight and control; however, the EU's current regulatory regime for genetically modified organisms (GMOs) is not necessarily conducive for stimulating development of new products. We therefore propose a differentiated regulatory framework that would considerably lower the regulatory hurdles for certain uses of genetic engineering, and stimulate innovation and development to the benefit of society, while allowing flexibility in terms of risk assessment. We further call upon relevant authorities and governments to advance policy development and facilitate a goal‐oriented and inclusive public dialogue. The aim is to harness the potential of gene technologies in a safe, beneficial, sustainable and ethically justifiable manner while maintaining public trust. ### The Current Situation In July 2018, after years of regulatory uncertainty in the EU, the European Court of Justice decided that all gene‐edited organisms are subject to the approval requirements that currently apply to the deliberate release of GMO under EU Directive 2001/18/EC [1]. The court's decision has been criticised by many European scientists [2], [3], [4], [5], who assert that it jeopardises the future of agriculture and food‐related research and development in the EU. In particular, many critics highlight that the current regulatory landscape will prevent the use of gene editing to rapidly develop novel agricultural products to deal …

Efficacy of a ML336 derivative against Venezuelan and eastern equine encephalitis viruses

Currently, there are no licensed human vaccines or antivirals for treatment of or prevention from infection with encephalitic alphaviruses. Because epidemics are sporadic and unpredictable, and endemic disease is common but rarely diagnosed, it is difficult to identify all populations requiring vaccination; thus, an effective post-exposure treatment method is needed to interrupt ongoing outbreaks. To address this public health need, we have continued development of ML336 to deliver a molecule with prophylactic and therapeutic potential that could be relevant for use in natural epidemics or deliberate release scenario for Venezuelan equine encephalitis virus (VEEV). We report findings from in vitro assessments of four analogs of ML336, and in vivo screening of three of these new derivatives, BDGR-4, BDGR-69 and BDGR-70. The optimal dosing for maximal protection was observed at 12.5 mg/kg/day, twice daily for 8 days. BDGR-4 was tested further for prophylactic and therapeutic efficacy in mice challenged with VEEV Trinidad Donkey (TrD). Mice challenged with VEEV TrD showed 100% and 90% protection from lethal disease when treated at 24 and 48 h post-infection, respectively. We also measured 90% protection for BDGR-4 in mice challenged with Eastern equine encephalitis virus. In additional assessments of BDGR-4 in mice alone, we observed no appreciable toxicity as evaluated by clinical chemistry indicators up to a dose of 25 mg/kg/day over 4 days. In these same mice, we observed no induction of interferon. Lastly, the resistance of VEEV to BDGR-4 was evaluated by next-generation sequencing which revealed specific mutations in nsP4, the viral polymerase.

Synthesis and Molecular Properties of Nerve Agent Reactivator HLö-7 Dimethanesulfonate.

The threat of a deliberate release of chemical nerve agents has underscored the need to continually improve field effective treatments for these types of poisonings. The oxime containing HLö-7 is a potential second-generation therapeutic reactivator. A synthetic process for HLö-7 is detailed with improvements to the DIBAL reduction and ion exchange steps. HLö-7 was visualized for the first time within the active site of human acetylcholinesterase and its relative ex vivo potency confirmed against various nerve agents using a phrenic nerve hemidiaphragm assay.

Development of a Biosecurity Checklist for Laboratory Assessment and Monitoring.

Introduction:Laboratory biosecurity is of continuously growing interest due to increasing concerns about deliberate misuse of biological materials and emerging biological risks. These risks continue to be magnified by globalization, the rapid pace of scientific development, and dual-use technologies. Worldwide laboratory capacities are expanding, which calls for concrete actions to improve laboratory biosafety and biosecurity practices to protect researchers and the community. Hence, laboratories require comprehensive biorisk management programs to minimize the risk of accidental and deliberate release of infectious biological materials.Objective:Malaysia has prioritized the concern of national biosecurity and aims to consolidate laboratory biosecurity performance to detect and prevent the deliberate release of biological agents.Methods:Two 3-day workshops were organized over the course of four months in which Malaysia collaborated with The Netherlands. This bilateral engagement aimed to integrate biosecurity practices in their national biorisk management programs, and resulted into a comprehensive biosecurity checklist for laboratory assessment and monitoring.Results:This biosecurity checklist is based on Malaysian and Dutch expert opinions and national and international guidelines and regulations. The biosecurity checklist is a survey-driven tool that consists of a set of concrete questions for each key biosecurity area, which are discussion points for assessment.Conclusion:We display a practical biosecurity checklist for laboratory assessment and monitoring. Although the presented checklist was the template for the specific Malaysia checklist, it could serve as a template for other countries.

The Court of Justice of the European Union’s Judgment on Mutagenesis and International Trade: A Case of GMO, Mutagenesis and International Trade: A Case of GMO, Mutagenesis and International Trad

The Court of Justice of the European Union (CJEU) was requested by the French Conseil d’État to determine, in essence, whether organisms obtained by mutagenesis (i.e. gene editing) are genetically modified organisms (GMOs) and whether they are subject to the obligations laid down by Directive 2001/18/EC of the European Parliament and of the Council of 12 March 2001 on the deliberate release into the environment of genetically modified organisms and repealing Council Directive 90/220/EEC. Contrary to the Advocate-General’s opinion of 18 January 2018, the Court of Justice ruled, on 25 July 2018, in case C-528/16 that Directive 2001/18/EC also applies to organisms obtained by mutagenesis techniques that have emerged since its adoption. The article analyses the grounds and implications of the CJEU’s decision. Unlike transgenesis, mutagenesis is a set of techniques, which make it possible to alter the genome of a living species without the insertion of foreign DNA. Indeed, it is virtually impossible to detect whether the DNA of a plant or animal has been edited or not, because the changes involved are indistinguishable from naturally occurring mutations. Mutagenesis techniques have made it possible to develop, inter alia, seed varieties that are resistant to selective herbicides. In response to the CJEU’s judgment, the US Secretary of Agriculture issued a statement criticizing the ruling. The statement reads, in part, that ‘[g]overnment policies should encourage scientific innovation without creating unnecessary barriers or unjustifiably stigmatizing new technologies. Unfortunately, this [...] ruling is a setback in this regard in that it narrowly considers newer genome editing methods to be within the scope of the European Union’s regressive and outdated regulations governing genetically modified organisms’. The article also analyses the possible international trade implications of this decision and of its effects, in light of the applicable WTO law and of the relevant case law, such as the EU – Biotech Products dispute by Argentina, Canada and the US, where the WTO had ultimately condemned a de facto moratorium on the approval of GM products in the EU and in its Member States.

Application of chlorine dioxide microcapsule sustained-release antibacterial films for preservation of mangos

In this study, fresh mangos were packed using a custom-made antimicrobial film coated with sustained-release chlorine dioxide microcapsules. We then compared physical and chemical indexes, such as weight loss rate, firmness, chromatic aberrations, soluble solids, vitamin C, titratable acid, and other nutritional indicators, to examine changes in the mango and film during storage. Our findings revealed that control mango showed loss of edible value and commercial value after 21days of storage, and the chlorine dioxide microcapsule antibacterial film group still retains food value and commercial value. Cross-sectional scanning electron microscopy images of the used film showed that the polylactic acid film was smooth and flat, whereas cross-sections of the antimicrobial film showed that the film was covered with voids due to deliberate release of chlorine dioxide gas during the packaging process. Thus, the antibacterial film exhibited erosion and degradation. These findings provided important insights into the use of antimicrobial films for the packaging of fruits during storage, which is essential for promoting the application of solid chlorine dioxide antimicrobial film in packaging preservation.

Exercise Mataika: White Paper on response to a smallpox bioterrorism release in the Pacific

Smallpox was declared eradicated in 1980, with known seed stock retained in two high security Biosafety Level 4 laboratories in the United States and Russia. Experts agree the likelihood of theft from these laboratories is low, and that synthetic creation of smallpox is a theoretical possibility. Until 2017 it was believed that synthetic smallpox was technically too complex a task to be a serious threat. However, in 2017, Canadian scientists synthesised a closely related orthopoxvirus, horsepox, using mail order DNA and $100,000. Simultaneously, terrorist groups have declared intent to conduct biological attacks. In this context an exercise was held on August 16th 2018, with international and cross-sectoral stakeholders to review preparedness for a bioterrorism attack in the Asia-Pacific region and globally. The exercise was conducted by The National Health and Medical Research Council (NHMRC) Centre for Research Excellence, Integrated Systems for Epidemic Response, with contextual input from the Ministry of Health and Medical Services Fiji. The scenario involved a deliberate release in Fiji, followed by a larger release in a more populous Asian country. Mathematical modelling was used to underpin epidemic projections under different conditions. The exercise alternated between clinical, public health, emergency and societal responses, with participants making real-time decisions on cross-sectoral response across the region and the world. Key weak points which are influential in determining the final size and impact of the epidemic were identified (based on mathematical modelling of transmission in Fiji and globally). We identified potential gaps in preparedness for smallpox and factors which influence the severity of a smallpox epidemic. This included identifying which determinants of epidemic size are potentially within our control, and which are not. Influential factors within our control include: preventing an attack through intelligence, law enforcement and legislation; speed of diagnosis; speed and completeness of case finding and case isolation; speed and security of vaccination response, including stockpiling; speed and completeness of contact tracing; protecting critical infrastructure and business continuity; non-pharmaceutical interventions (social distancing, PPE, border control); protecting first responders; operational support and logistics; social mobilisation and risk communication. Based on discussion at the workshop between diverse stakeholders, recommendations were made to guide improved prevention, mitigation and rapid response, thus providing a holistic, cross-sectoral framework for prevention of a worst-case scenario smallpox pandemic.

Essential evidence-based introductory bioterrorism content for practicing nurses.

Bioterrorism content is too often absent from nursing education post-licensure. According to the Centers for Disease Control and Prevention (CDC), bioterrorism is defined as "the deliberate release of viruses, bacteria, or other germs (agents) used to cause illness or death in people, animals, or plants." The purpose of this scholarly work was to develop and pilot a curriculum on introductory bioterrorism concepts for practicing nurses. A literature search and a review of available resources elucidated pertinent content items on bioterrorism and its associated precautions for nurses. Next, two rounds of expert panel consisting of five members rated the identified content and provided comments to validate the curriculum's content. Based on the results, content items were expanded into a curriculum that was piloted as a four hour in person educational session and nurses in attendance were invited to participate in pre-test and post-test questions. They were also asked for their perceptions of the educational session's usefulness and influence on patient care in case of a bioterrorism event. All respondents strongly agreed that the information presented would be useful to them and would influence their patient care in a bioterrorism event. Further refinement, evaluation, and implementation of the developed curriculum are recommended.

Effects of Alternative Blood Sources on Wolbachia Infected Aedes aegypti Females within and across Generations.

Wolbachia bacteria have been identified as a tool for reducing the transmission of arboviruses transmitted by Aedes aegypti. Research groups around the world are now mass rearing Wolbachia-infected Ae. aegypti for deliberate release. We investigated the fitness impact of a crucial element of mass rearing: the blood meal required by female Ae. aegypti to lay eggs. Although Ae. aegypti almost exclusively feed on human blood, it is often difficult to use human blood in disease-endemic settings. When females were fed on sheep or pig blood rather than human blood, egg hatch rates decreased in all three lines tested (uninfected, or infected by wMel, or wAlbB Wolbachia). This finding was particularly pronounced when fed on sheep blood, although fecundity was not affected. Some of these effects persisted after an additional generation on human blood. Attempts to keep populations on sheep and pig blood sources only partly succeeded, suggesting that strong adaptation is required to develop a stably infected line on an alternative blood source. There was a decrease in Wolbachia density when Ae. aegypti were fed on non-human blood sources. Density increased in lines kept for multiple generations on the alternate sources but was still reduced relative to lines kept on human blood. These findings suggest that sheep and pig blood will entail a cost when used for maintaining Wolbachia-infected Ae. aegypti. These costs should be taken into account when planning mass release programs.

Sampling and inactivation of wet disseminated spores from flooring materials, using commercially available robotic vacuum cleaners.

Four commercially available robotic vacuum cleaners were assessed for sampling efficiency of wet disseminated Bacillus atrophaeus spores on carpet, polyvinyl chloride (PVC) and laminate flooring. Furthermore, their operability was evaluated and decontamination efficiency of one robot was assessed, using a sodium hypochlorite solution. In an environmental chamber, robots self-navigated around 4m2 of flooring containing a single contaminated 0·25m2 tile (c.104 spores per cm2 ). Contamination levels at predetermined locations were assessed by macrofoam swabs (PVC and laminate) or water soluble tape (carpet), before and after sampling. Robots were dismantled postsampling and spore recoveries assessed. Aerosol contamination was also measured during sampling. Robot sampling efficiencies were variable, however, robots recovered most spores from laminate (up to 17·1%), then PVC and lastly the carpet. All robots spread contamination from the 'hotspot' (all robots spread <0·6% of the contamination to other areas) and became surface contaminated. Spores were detected at low levels during air sampling (<5·6 spores per litre). Liquid decontamination inactivated 99·1% of spores from PVC. Robotic vacuum cleaners show promise for both sampling and initial decontamination of indoor flooring. In the event of a bioterror incident, e.g. deliberate release of Bacillus anthracis spores, areas require sampling to determine the magnitude and extent of contamination, and to establish decontamination efficacy. In this study, we investigate robotic sampling methods against high concentrations of bacterial spores applied by wet deposition to different floorings, contamination spread to other areas, potential transfer of spores to the operators and assessment of a wet vacuum robot for spore inactivation. The robots' usability was evaluated and how they can be employed in real life scenarios. This will help to reduce the economic cost of sampling and the risk to sampling/decontamination teams.

Sensitive and Specific Recombinase Polymerase Amplification Assays for Fast Screening, Detection, and Identification of Bacillus anthracis in a Field Setting

ABSTRACTFour isothermal recombinase polymerase amplification (RPA) assays were developed for fast in-field identification of Bacillus anthracis. The RPA assays targeted three specific sequences (i.e., the BA_5345 chromosomal marker, the lethal factor lef [from pXO1], and the capsule-biosynthesis-related capA [from pXO2]) and a conserved sequence in the adenylate cyclase gene (adk) for the Bacillus cereus group. B. anthracis-specific RPA assays were tested first with purified genomic DNAs (n = 60), including 11 representatives of B. anthracis, and then with soil (n = 8) and white powder (n = 8) samples spiked with inactivated B. anthracis spores and/or other biological agents. The RPA assays were also tested in another laboratory facility, which blindly provided DNA and lysate samples (n = 30, including 20 B. anthracis strains). RPA assays displayed 100% specificity and sensitivity. The hands-off turnaround times at 42°C ranged from 5 to 6 min for 102 genomic copies. The analytical sensitivity of each RPA assay was ∼10 molecules per reaction. In addition, the BA_5345 and adk RPA assays were assessed under field conditions with a series of surface swabs (n = 13, including 11 swabs contaminated with B. thuringiensis spores) that were blindly brought to the field laboratory by a chemical, biological, radiological, and nuclear (CBRN) sampling team. None of the 13 samples, except the control, tested positive for B. anthracis, and all samples that had been harvested from spore-contaminated surfaces tested positive with the adk RPA assay. All three B. anthracis-specific RPA assays proved suitable for rapid and reliable identification of B. anthracis and therefore could easily be used by first responders under field conditions to quickly discriminate between a deliberate release of B. anthracis spores and a hoax attack involving white powder.IMPORTANCE In recent decades, particularly following the 11 September 2001 and Amerithrax attacks, the world has experienced attempts to sow panic and chaos in society through thousands of white-powder copycats using household powders to mimic real bioterrorism attacks. In such circumstances, field-deployable detection methods are particularly needed to screen samples collected from the scene. The aim is to test the samples directly using a fast and reliable assay for detection of the presence of B. anthracis. While this would not preclude further confirmatory tests from being performed in reference laboratories, it would bring useful, timely, and relevant information to local crisis managers and help them make appropriate decisions without having to wait for quantitative PCR results (with turnaround times of a few hours) or phenotypic identification and sequencing (with turnaround times of a few days). In the current investigation, we developed a set of isothermal RPA assays for the rapid screening and identification of B. anthracis in powders and soil samples, with the purpose of discriminating a deliberate release of B. anthracis spores from a hoax attack involving white powder; this would also apply to dispersion by spraying of aerosolized forms of B. anthracis. Further work is now ongoing to confirm the first observations and validate the on-site use of these assays by first responders.

Butyrated ManNAc analog improves protein expression in Chinese hamster ovary cells.

The chemical additive sodium butyrate (NaBu) has been applied in cell culture media as a direct and convenient method to increase the protein expression in Chinese hamster ovary (CHO) and other mammalian cells. In this study, we examined an alternative chemical additive, 1,3,4-O-Bu3 ManNAc, for its effect on recombinant protein production in CHO. Supplementation with 1,3,4-O-Bu3 ManNAc for two stable CHO cell lines, expressing human erythropoietin or IgG, enhanced protein expression for both products with negligible impact on cell growth, viability, glucose utilization, and lactate accumulation. In contrast, sodium butyrate treatment resulted in a ∼20% decrease in maximal viable cell density and ∼30% decrease in cell viability at the end of cell cultures compared to untreated or 1,3,4-O-Bu3 ManNAc treated CHO cell lines for both products. While NaBu treatment enhanced product yields more than the 1,3,4-O-Bu3 ManNAc treatment, the NaBu treated cells also exhibited higher levels of caspase 3 positive cells using microscopy analysis. Furthermore, the mRNA levels of four cell apoptosis genes (Cul2, BAK, BAX, and BCL2L11) were up-regulated more in sodium butyrate treated wild-type, erythropoietin, or IgG expressing CHO-K1 cell lines while most of the mRNA levels of apoptosis genes in 1,3,4-O-Bu3 ManNAc treated cell lines remained equal or increased only slightly compared to the levels in untreated CHO cell lines. Finally, lectin blot analysis revealed that the 1,3,4-O-Bu3 ManNAc-treated cells displayed higher relative sialylation levels on recombinant EPO, consistent with the effect of the ManNAc component of this additive, compared to control while NaBu treatment led to lower sialylation levels than control, or 1,3,4-O-Bu3 ManNAc-treatment. These findings demonstrate that 1,3,4-O-Bu3 ManNAc has fewer negative effects on cell cytotoxicity and apoptosis, perhaps as a result of a more deliberate uptake and release of the butyrate compounds, while simultaneously increasing the expression of multiple recombinant proteins, and improving the glycosylation characteristics when applied at comparable molarity levels to NaBu. Thus, 1,3,4-O-Bu3 ManNAc represents a highly promising media additive alternative in cell culture for improving protein yields without sacrificing cell mass and product quality in future bioproduction processes.

The Impact of Release Management on Open-Source Software Co-Creation

The open-source software (OSS) production model has been gaining ground and even expanding to a broader class of products. A central virtue of OSS is co-creation through contributions and feedback from the user community, yet our knowledge of how to coordinate and maximize the benefit of such co-creation for market success is limited. In this paper, we propose that deliberate product release timing can be an important capability to orchestrate open source community contributions and maximize their benefit. We develop, formalize, and estimate a dynamic structural model to study the impact of product release management as a coordinating mechanism in peer production. We find that a deliberate and moderate release frequency, contingent on the installed base of the project, its quality enhancement, and the license type of the software, contributes to project success. Our results show that it would be optimal for early-stage projects to release more often. As the installed base increases, it is beneficial to release less often. We also find that there exists a curvilinear (inverse-U) relationship between the release frequency and the community contributions. Hence, excessive releasing may risk exhausting the community. Furthermore, even though the project may benefit from a new release, there also exists a release cost. The OSS team then has to balance between the two by carefully managing the release pace. We also find that projects with different license types may have different optimal release timing. These results may have important implications for managing technology-enabled, crowd-based collaboration in open innovation communities beyond open-source software development, launch, and adoption.

Civilian exposure to chlorine gas: A systematic review

IntroductionHalogen pulmonary irritants (HPIs) are volatile liquids that directly damage the respiratory mucosa. Chlorine is readily available in large volumes as an industrial chemical and has a significant potential for accidental or deliberate release. We conducted a systematic review to determine the clinical features; treatment and long-term sequelae of civilian chlorine gas exposure. MethodsA systematic review was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) methodology. Medline; Ovid and Google Scholar databases were searched from 1966 to January 2017. A database of relevant papers was compiled and descriptive statistics used to summarise the data. ResultsThirty-six papers describing 37 incidents involving 1566 individual acute exposers to chlorine gas were identified. The most common reported features were cough (29%), dyspnoea (22%), sore throat (16%), eye features (12%) and excessive sputum or haemoptysis (7%). Acute management included high-flow oxygen (32.8%); steroids (28.4%); bronchodilators (28.2%) and ventilation (2.3%). Nine deaths (0.6%) were reported. Follow-up data available in 60% of cases; full recovery was reported in 90% of cases where data was available. DiscussionAcute chlorine gas exposure in civilian incidents presented with acute respiratory features and irritation of the eyes and throat. The development of pulmonary oedema or ARDS was relatively rare when compared to military experience in the First World War.

Expression of haloacid dehalogenase gene and its molecular protein characterization from Klebsiella pneumoniae ITB1

Organohalogen compounds are widely used industrially and agriculturally, as well as in households as flame retardants and refrigerants. However, these compounds can become significant pollutants through their accidental or deliberate release into the environment in large quantities. Dehalogenase is an enzyme with the potential to be used in the removal of organohalogen contaminants. A previous study successfully subcloned a 690 bp of haloacid dehalogenase gene (hakp1) from Klebsiella pneumoniae ITB1 into a pET-30a(+) expression system to achieve high enzyme productivity. IPTG was used as an inducer to express a pET-hakp1 recombinant clone in Escherichia coli BL21 (DE3). The molecular mass of the haloacid dehalogenase Hakp1 protein was 30 kDa as determined by SDS-PAGE. Zymogram analysis showed that this recombinant protein has dehalogenase activity as shown by the formation of AgCl white precipitate. A quantitative assay of haloacid dehalogenase Hakp1 gave a specific activity of 84.29 U/mg with the optimum temperature of 40°C at pH 9. Predicted three-dimensional structure of Hakp1 showed α/β motif folding which comprised of cap and core domain. The predicted active sites of Hakp1 were Asp8, Glu10, Leu22, Phe23, Trp90, Ser125, Ser126, Lys159, and Asp184 with Asp8, Glu10, Ser126, and Lys159 act as binding residue. This recombinant haloacid dehalogenase clone provides an alternative agent for effective bioremediation of organohalogen pollutants.

Developing gene drive technologies to eradicate invasive rodents from islands

ABSTRACTIsland ecosystems are highly threatened by invasive rats and mice. Currently, the only effective technology for eradicating rodents from islands is toxicants. Though effective, they are expensive and have high failure rates. They are not species-specific and are potentially dangerous to humans. Gene drive technology is one alternative to toxicants for rodent eradication. Gene drive methods of rodent eradication offer an alternative to killing that has the potential to be more species-specific, more humane, and more biologically safe for use around humans. Technologies in development aim to apply either natural meiotic drive or clustered regularly interspersed short palindromic repeats to influence offspring development so that all offspring are phenotypically male, eventually creating a population that is not reproductively viable. Implementing this technology would involve releasing laboratory-developed engineered mice into wild populations. Some areas for further research include assessing the ecological effects of releasing engineered mice, the potential risks for the accidental or deliberate release of genetically modified organisms into mainland mouse populations, and the social, ethical, and regulatory acceptability of the technology.

Continuous positive airway pressure: An early intervention to prevent phosgene-induced acute lung injury

Exposure to toxic industrial chemicals such as phosgene may occur through accidental or deliberate release. Inhalation may result in an acute lung injury which manifests as hypoxaemia with insufficient oxygen being delivered to the tissues resulting in hypoxia, respiratory failure and death. No effective pharmacological therapy currently exists and treatment remains supportive, often requiring intensive care facilities. In a mass casualty scenario the logistical burden of managing exposed individuals would rapidly overwhelm healthcare systems. This highlights the need to develop post exposure therapeutic strategies to minimise injury severity and increase survival in individuals exposed to toxic chemicals.Our research objective was to investigate a commercial off the shelf (COTS) therapy; ambient air continuous positive airway pressure (CPAP) support, initiated 1h post exposure to explore the concept that early intervention with positive airway pressure would reduce or ameliorate lung injury following exposure to phosgene.This study has demonstrated that CPAP, initiated before overt signs of exposure become manifest, significantly improved survival as well as improving some clinically relevant physiological measures of phosgene-induced acute lung injury over 24h.

A Systematic Review of Risk Analysis Tools for Differentiating Unnatural From Natural Epidemics

ABSTRACTIntroduction: In the era of genetic engineering of pathogens, distinguishing unnatural epidemics from natural ones is a challenge. Successful identification of unnatural infectious disease events can assist in rapid response, which relies on a sensitive risk assessment tool used for the early detection of deliberate attacks (i.e., bioterrorism). Methods: A systematic review was conducted according to the outline of Preferred Reporting Items for Systematic Reviews. Published papers related to the detection of unnatural diseases were searched in MEDLINE (January 1927–April 2016), EMBASE (January 1937–March 2016), and Web of Science (January 1978–March 2016). Full texts were reviewed for the selection of studies on scoring systems specially designed to discern between unnatural and natural outbreaks. Results: A total of 1,753 papers were reviewed, of which we identified the following five scoring systems specifically designed for detecting unnatural outbreaks: (1) the Grunow–Finke epidemiological assessment tool, (2) potential epidemiological clues to a deliberate epidemic, (3) bioterrorism risk assessment scoring, (4) and (5) two modified scoring systems based on (3). Various criteria ranging from the information on perpetrators, type of agents, spatial distribution, and intelligence of deliberate release were involved. Of these systems, the Grunow–Finke assessment tool remains the most widely used, but has low sensitivity for correctly identifying unnatural epidemics when tested against actual historical outbreaks. Others were applied into a few scenarios but provided different perspectives for bioterrorism detection and bio-preparedness. Conclusion: There are few risk assessment tools for differentiating unnatural from natural epidemics. These tools are increasingly necessary and valuable, but improved scoring systems with higher sensitivity, specificity, timeliness, and wider application to biological attacks must be developed.