Acetoacetate and ethyl acetoacetate as novel inhibitors of bacterial biofilm.

Acetoacetate (AAA) was identified as a biofilm inhibitor in a previous study, where the effect of 190 carbon and nitrogen sources on biofilm amounts by Escherichia coli O157:H7 was determined. With this study, we tested the effect of AAA on growth and biofilm amounts of Cronobacter sakazakii, Serratia marcescens and Yersinia enterocolitica. AAA reduced growth and biofilm amounts of the three pathogens, albeit at rather high concentrations of 10 to 35 mg ml-1 . Acetoacetate at a concentration of 5 mg ml-1 reduced Y. enterocolitica mRNA transcripts of the flagellar master regulator operon flhD, the invasion gene inv, and the adhesion gene yadA. Transcription of the regulator of plasmid-encoded virulence genes virF, the plasmid-encoded virulence gene yopQ, and ymoA were largely unaffected by AAA. Importantly, AAA did not cause an increase in transcription of any of the tested virulence genes. As a more cost efficient homologue of AAA, the effect of ethyl acetoacetate (EAA) was tested. EAA reduced growth, biofilm amounts and live bacterial cell counts up to 3 logs. IC50 values ranged from 0·31 mg ml-1 to 5·6 mg ml-1 . In summary, both AAA and EAA inhibit biofilm, but EAA appears to be more effective. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacterial biofilms are communities of bacteria that form on surfaces and are extremely difficult to remove by conventional physical or chemical techniques, antibiotics or the human immune system. Despite advanced technologies, biofilm still contributes to 60 to 80% of human bacterial infections (NIH and CDC) and cause problems in many natural, environmental, bioindustrial or food processing settings. The discovery of novel substances that inhibit biofilm without increasing the virulence of the bacteria opens doors for countless applications where a reduction of biofilm is desired.

The prospective validation of the Modified Physiological Triage Tool (MPTT): an evidence-based approach to major incident triage.

INTRODUCTION: Triage is a key principle in the effective management of major incidents. There is limited evidence to support existing triage tools, with a number of studies demonstrating poor performance at predicting the need for a life-saving intervention. The Modified Physiological Triage Tool (MPTT) is a novel triage tool derived using logistic regression, and in retrospective data sets has shown optimum performance at predicting the need for life-saving intervention. MATERIALS AND METHODS: Physiological data and interventions were prospectively collected for consecutive adult patients with trauma (>18 years) presenting to the emergency department at Camp Bastion, Afghanistan, between March and September 2011. Patients were considered priority 1 (P1) if they received one or more interventions from a previously defined list. Patients were triaged using existing triage tools and the MPTT. Performance was measured using sensitivity and specificity, and a McNemar test with Bonferroni calculation was applied for tools with similar performance. RESULTS: The study population comprised 357 patients, of whom 214 (59.9%) were classed as P1. The MPTT (sensitivity: 83.6%, 95% CI 78.0% to 88.3%; specificity: 51.0%, 95% CI 42.6% to 59.5%) outperformed all existing triage tools at predicting the need for life-saving intervention, with a 19.6% absolute reduction in undertriage compared with the existing Military Sieve. The improvement in undertriage comes at the expense of overtriage; rates of overtriage were 11.6% higher with the MPTT than the Military Sieve. Using a McNemar test, a statistically significant (p<0.001) improvement in overall performance was demonstrated, supporting the use of the MPTT over the Military Sieve. DISCUSSION AND CONCLUSIONS: The MPTT outperforms all existing triage tools at predicting the need for life-saving intervention, with the lowest rates of undertriage while maintaining acceptable levels of overtriage. Having now been validated on both military and civilian cohorts, we recommend that the major incident community consider adopting the MPTT for the purposes of primary triage.

Predictors of massive blood transfusion: a Delphi Study to examine the views of experts.

BACKGROUND: Trauma patients requiring massive blood transfusion (MBT) have high morbidity and mortality: early and aggressive use of blood products during immediate resuscitation may improve survival. There is currently a lack of evidence to guide initial identification of these patients which is especially important in areas where plasma may need to be thawed. In the absence of this evidence, this study aimed to robustly evaluate expert opinion by using a Delphi process to identify predictors of massive transfusion. This process can be used to ensure that decision rules include variables that have clinical validity, which may improve translation of rules into clinical practice. METHODS: An international panel of 35 experts was identified through expert advice against specific criteria. Military and civilian experts from the fields of emergency medicine, critical care, anaesthesia, prehospital care, haematology and general/trauma surgery were included. The Delphi Study was carried out over three rounds. Consensus level was predefined at 80%. RESULTS: 195 statements were generated by the panel of which 97 (49.7%) achieved consensus at the 80% level by the end of round 3. Strikingly no clinical observations reached consensus individually. Metabolic acidosis of a base excess of -5.0 or worse, lactate >5 mmol/L and a low haematocrit on arrival were all considered predictive. Some patterns of injury, but few mechanisms of injury, were considered highly predictive of the need of MBT. CONCLUSIONS: This Delphi process has produced a list of parameters that expert clinicians felt were predictive for MBT. This list can be used to inform the generation of decision rules. It is of note that many factors used in current decision rules were not valued by clinical experts-this may be a cause for poor uptake of those rules.

Recurrent gain-of-function mutation in PRKG1 causes thoracic aortic aneurysms and acute aortic dissections.

Gene mutations that lead to decreased contraction of vascular smooth-muscle cells (SMCs) can cause inherited thoracic aortic aneurysms and dissections. Exome sequencing of distant relatives affected by thoracic aortic disease and subsequent Sanger sequencing of additional probands with familial thoracic aortic disease identified the same rare variant, PRKG1 c.530G>A (p.Arg177Gln), in four families. This mutation segregated with aortic disease in these families with a combined two-point LOD score of 7.88. The majority of affected individuals presented with acute aortic dissections (63%) at relatively young ages (mean 31 years, range 17-51 years). PRKG1 encodes type I cGMP-dependent protein kinase (PKG-1), which is activated upon binding of cGMP and controls SMC relaxation. Although the p.Arg177Gln alteration disrupts binding to the high-affinity cGMP binding site within the regulatory domain, the altered PKG-1 is constitutively active even in the absence of cGMP. The increased PKG-1 activity leads to decreased phosphorylation of the myosin regulatory light chain in fibroblasts and is predicted to cause decreased contraction of vascular SMCs. Thus, identification of a gain-of-function mutation in PRKG1 as a cause of thoracic aortic disease provides further evidence that proper SMC contractile function is critical for maintaining the integrity of the thoracic aorta throughout a lifetime.

Coordination and relationships between organisations during the civil-military international response against Ebola in Sierra Leone: an observational discussion.

The Ebola virus disease (EVD) crisis in West Africa began in March 2014. At the beginning of the outbreak, no one could have predicted just how far-reaching its effects would be. The EVD epidemic proved to be a unique and unusual humanitarian and public health crisis. It caused worldwide fear that impeded the rapid response required to contain it early. The situation in Sierra Leone (SL) forced the formation of a unique series of civil-military interagency relationships to be formed in order to halt the epidemic. Civil-military cooperation in humanitarian situations is not unique to this crisis; however, the slow response, the unusual nature of the battle itself and the uncertainty of the framework required to fight this deadly virus created a situation that forced civilian and military organisations to form distinct, cooperative relationships. The unique nature of the Ebola virus necessitated a steering away from normal civil-military relationships and standard pillar responses. National and international non-governmental organisations (NGOs), Department for International Development (DFID) and the SL and UK militaries were required to disable this deadly virus (as of 7 November 2015, SL was declared EVD free). This paper draws on personal experiences and preliminary distillation of information gathered in formal interviews. It discusses some of the interesting features of the interagency relationships, particularly between the military, the UK's DFID, international organisations, NGOs and departments of the SL government. The focus is on how these relationships were key to achieving a coordinated solution to EVD in SL both on the ground and within the larger organisational structure. It also discusses how these relationships needed to rapidly evolve and change along with the epidemiological curve.

Op GRITROCK ethics; the way of things to come?

The Defence Medical Services (DMS) deployed on Op GRITROCK to Sierra Leone in support of the Ebola outbreak. This operation was the first large-scale DMS deployment since operations in Afghanistan ceased at the end of 2014. This type of operation revealed a number of ethical issues and challenges that the DMS had not faced for a long time. The ethical issues identified during the deployment are discussed in this article using the principalism approach of Beauchamp and Childress. Many of these issues were not identified prior to deployment, and troops were not adequately prepared. The article will outline the difficulties of using the principles of autonomy, beneficence, non-maleficence and justice during a public health emergency, and conclude with recommendations for ethical considerations that should be identified and mitigated against for future deployments to a public health emergency.

Usefulness of the Shock Index as a secondary triage tool.

INTRODUCTION: Secondary triage at a major incident allows for a more detailed assessment of the patient. In the UK, the Triage Sort (TSO) is the preferred method, combining GCS, systolic BP (SBP) and RR to categorise Priority 1 casualties. The Shock Index (SI) is calculated by dividing HR by SBP (HR/SBP). This study examines whether SI is better at predicting need for life-saving intervention (LSI) following trauma than TSO. METHODS: A prospective observational study was undertaken. Physiological data and interventions performed in the Emergency Department and operating theatre were prospectively collected for 482 consecutive adult trauma patients presenting to Camp Bastion, Afghanistan, over a 6-month period. A patient was deemed to have required LSI if they received any intervention from a set described previously. RESULTS: Complete data were available for 345 patients (71.6%). Of these, 203 (58.8%) were gold standard P1, and 142 (41.2%) were non-P1. The TSO predicted need for LSI with a sensitivity of 58.6% (95% CI 51.8% to 65.4%) and specificity of 88.7% (95% CI 83.5% to 93.9%). Using an SI cut-off >0.75 provided greater sensitivity of 70.0% (95% CI 63.6% to 76.3%) while maintaining an acceptably high (although lower than TSO) specificity of 74.7% (95% CI 67.5% to 81.8%). At this SI cut-off, there was evidence of a difference between TSO and SI in terms of the way in which patients were triaged (p<0.0001). DISCUSSION: Our study showed that a SI >0.75 more accurately predicted the need for LSI, while maintaining acceptable specificity. SI may be more useful than TSO for secondary triage in a mass-casualty situation; this relationship in civilian trauma should be examined to clarify whether these results can be more widely translated into civilian practice. PROJECT REGISTRATION NUMBER: RCDM/Res/Audit/1036/12/0050.

Cervical spine clearance in the elderly: do elderly patients get a bad deal?

OBJECTIVE: To investigate whether elderly patients, presenting following trauma and requiring immobilisation and imaging of the cervical spine, require increased use of CT and spend longer immobilised when compared to the younger population. METHODS: A retrospective chart review was undertaken of 35 adults aged 18-65 years, and 32 adults over 65 years, requiring cervical spine imaging following trauma. RESULTS: 1 of the 35 younger patients, and 16 of the 32 elderly patients, underwent CT. Elderly patients spent 1:05 h longer immobilised (p<0.005). CONCLUSIONS: Half of the elderly patients underwent CT, and they were immobilised for significantly longer than younger patients. Measures should be adopted to facilitate early diagnosis and mitigate complications of prolonged immobilisation in elderly patients with neck trauma.

Intraosseous access in the military operational setting.

INTRODUCTION: In an operational military environment, circulatory access can prove difficult for a variety of reasons including profound hypovolaemia, and limited first responder experience. With many injuries that cause catastrophic haemorrhage, such as traumatic limb amputations, circulatory access is needed as quickly as possible. Since 2006, the Defence Medical Services have been using the EZ-IO and FAST1 devices as a means of obtaining circulatory access. METHODS: A prospective observational study was conducted between March and July 2011 at the Emergency Department, Camp Bastion, Afghanistan. All patients with an intraosseous device had data recorded that included if the device successfully flushed (functionality) and if any problems were encountered. RESULTS: 117 patients presented with a total of 195 devices: 149 were EZ-IO (76%) and 46 were FAST1 (24%). Functionality was recorded for 111 (57%), with 17 failing to function, yielding an overall success rate of 84.7%. Device failure was observed to be more prevalent in the humerus; inability to flush the device was the leading cause, followed by mechanical failure. There were 2 complications, device breaking on removal being the reason for both. CONCLUSIONS: The devices in the study were tested for a period of time following insertion (median 32 minutes), and still the success rates mirror those found in the literature. Observed differences between sites were not found to be significant with confidence intervals overlapping. Further work is proposed to investigate the long-term complications of intraosseous devices.

Operational patient care pathway: building pragmatic medical interoperability through health engagements.

The future operational demand for medical support in Western militaries will likely outstrip available resources, necessitating burden-sharing through medical interoperability with allies and partners. However, the current North Atlantic Treaty Organization (NATO) model of interoperability through standardisation, while achieving high levels of commonality and integration along the operational patient care pathway (OPCP), is high-cost and resource-intensive. We have termed this model assured interoperability. Assured interoperability, while applicable to well-established partnerships with high-resource nations, is unlikely to be feasible when working with resource-limited partners or, potentially, when in a sustained conflict with a near-peer adversary. In these circumstances, there will be a requirement to develop a far less resource-intensive model of medical interoperability with lower levels of commonality, assurance and standardisation than assured interoperability, but that provides a 'good enough' OPCP for the operational context. We have termed this pragmatic interoperability. By considering these two types of interoperability, the complete continuum of medical interoperability can be mapped with the full spectrum of partners demonstrating increasing levels of interoperability from pragmatic through to assured interoperability, integrateability and interchangeability, reducing the gap between demand and provision of medical support for operations, increasing operational resilience. This is a paper commissioned as a part of the Defence Engagement special issue of BMJ Military Health.

Karyotype heterogeneity and unclassified chromosomal abnormalities.

In a departure from traditional gene-centric thinking with regard to cytogenetics and cytogenomics, the recently introduced genome theory calls upon a re-focusing of our attention on karyotype analyses of disease conditions. Karyotype heterogeneity has been demonstrated to be directly involved in the somatic cell evolution process which is the basis of many common and complex diseases such as cancer. To correctly use karyotype heterogeneity and apply it to monitor system instability, we need to include many seemingly unimportant non-specific chromosomal aberrations into our analysis. Traditionally, cytogenetic analysis has been focused on identifying recurrent types of abnormalities, particularly those that have been linked to specific diseases. In this perspective, drawing on the new framework of 4D-genomics, we will briefly review the importance of studying karyotype heterogeneity. We have also listed a number of overlooked chromosomal aberrations including defective mitotic figures, chromosome fragmentation as well as genome chaos. Finally, we call for the systematic discovery/characterization and classification of karyotype abnormalities in human diseases, as karyotype heterogeneity is the common factor that is essential for somatic cell evolution.

Heterogeneity of cell death.

Cell death constitutes a number of heterogeneous processes. Despite the dynamic nature of cell death, studies of cell death have primarily focused on apoptosis, and cell death has often been viewed as static events occurring in linear pathways. In this article we review cell death heterogeneity with specific focus on 4 aspects of cell death: the type of cell death; how it is induced; its mechanism(s); the results of cell death, and the implications of cell death heterogeneity for both basic and clinical research. This specifically reveals that cell death occurs in multiple overlapping forms that simultaneously occur within a population. Network and pathway heterogeneity in cell death is also discussed. Failure to integrate cell death heterogeneity within analyses can lead to inaccurate predictions of the amount of cell death that takes place in a tumor. Similarly, many molecular methods employed in cell death studies homogenize a population removing heterogeneity between individual cells and can be deceiving. Finally, and most importantly, cell death heterogeneity is linked to the formation of new genome systems through induction of aneuploidy and genome chaos (rapid genome reorganization).

Ultrasound of penetrating ocular injury in a combat environment.

Penetrating eye injuries are uncommon in a civilian environment, but unfortunately, very common in a military emergency department. Ultrasound of the eye is quick, reliable, accurate, and easy to learn. This review aims to demonstrate normal anatomy and penetrating injuries of the anterior and posterior compartments of the eye.

Vertebrate genome evolution and the zebrafish gene map.

In chordate phylogeny, changes in the nervous system, jaws, and appendages transformed meek filter feeders into fearsome predators. Gene duplication is thought to promote such innovation. Vertebrate ancestors probably had single copies of genes now found in multiple copies in vertebrates and gene maps suggest that this occurred by polyploidization. It has been suggested that one genome duplication event occurred before, and one after the divergence of ray-finned and lobe-finned fishes. Holland et al., however, have argued that because various vertebrates have several HOX clusters, two rounds of duplication occurred before the origin of jawed fishes. Such gene-number data, however, do not distinguish between tandem duplications and polyploidization events, nor whether independent duplications occurred in different lineages. To investigate these matters, we mapped 144 zebrafish genes and compared the resulting map with mammalian maps. Comparison revealed large conserved chromosome segments. Because duplicated chromosome segments in zebrafish often correspond with specific chromosome segments in mammals, it is likely that two polyploidization events occurred prior to the divergence of fish and mammal lineages. This zebrafish gene map will facilitate molecular identification of mutated zebrafish genes, which can suggest functions for human genes known only by sequence.

Personal view: security sector health systems and global health.

In many countries the security sector is a major contributor to the healthcare system. The role and transformation of a state's security health system within the universal health coverage is important, in that it sits at the interface of the United Nations Sustainable Development Goals 3 (ensure healthy lives and promote well-being) and 16 (promote just, peaceful and inclusive societies). The paper describes the breadth of the security sector and outlines the potential beneficiaries, clinical services and macro-organisation of a security sector health system from the perspective of its contribution to wider government health services and crisis response. It examines the characteristics of the security sector compared with other providers of health services, including those generic to the sector and unique to a given service. Understanding civil-security relationships is a critical facet of effective Defence Healthcare Engagement (DHE), which includes the use of defence medical assets in support of capacity-building overseas. The analytical process described may form the basis of DHE planning. It may have even greater importance in the near future as countries review national resilience and global health diplomacy after the COVID-19 crisis.

Fellowships and defence engagement: from clinical niches to strategic impact.

Medical fellowships have traditionally developed the individual rather than furthering military or national strategic objectives. This paper describes a medical fellowship with the British Antarctic Survey to illustrate the benefits to the individual, to the military and to wider international defence engagement efforts.By rebranding such fellowships as Defence Healthcare Engagement and by treating international organisations on a par with partner nations, the humble fellowship can facilitate enduring, mutually beneficial healthcare engagement at low cost and with minimal additional resources.

Medics as influencers: a historical analysis of British Army military medical exercises in Kenya over two decades.

INTRODUCTION: Annual British Army medical training exercises have run in Kenya since the early 1990s, initially with a dual purpose-to deliver the Kenyan Extended Programme of Immunisation (in remote locations) and to undertake austere training. This provided a specific response to a capability gap request from the partner nation, but as this gap closed, the exercise changed in various ways. This study aimed to qualitatively explore the impact of these exercises on the Kenyan healthcare system and the influence and relationships between the nations. METHODS: Semistructured interviews were conducted for 10 former senior commanders and medical officers who had deployed in key command and clinical positions from 1993 to 2019. Three researchers conducted thematic content analysis on the key-informant interviews. RESULTS: Five domains with 18 subdomains formed the study's analysis framework. 16 recurring themes were identified and placed into four categories that denote if they were of benefit to the engagement, enabled success, had the potential to cause harm or were a barrier to successful engagement. Three distinct phases of the exercise were identified: supporting Kenyan vaccinations, direct clinical care, training and education. CONCLUSIONS: This is the first qualitative analysis of the impact of a British Defence Engagement (Health) on the partner nation and UK influence gained through it. It has identified factors which may improve outcomes, namely, ensuring sustainability and continuity between iterations; maintaining enduring stakeholder relationships; responding to a capability gap request; intelligence-led planning with incorporated assessment, monitoring and evaluation; adapting to changes in needs or contextual settings; while ensuring mutual benefit in objective setting. These may be used as the basis for a conceptual framework supporting the planning and execution of high-quality, mutually beneficial Defence Engagement (Health) activities in future. This framework and future research would also benefit from gaining perspectives from the partner nation.

Defence context for the UK's Defence Engagement (Health).

Defence Engagement (DE) has been a core UK Defence task since 2015. DE (Health) is the use of military medical capabilities to achieve DE effects within the health sector to achieve security and defence objectives. DE (Health) practitioners must understand the underlying defence context that shapes these objectives. The strategic context is becoming more uncertain with the return of great power competition layered on enduring threats from non-state actors and transnational challenges. The UK response has been to develop the Integrated Review, outlining four national security and international policy objectives. UK Defence has responded by developing the integrated operating concept, differentiating military activity between operating and warfighting. Engage is one of the three functions of operate activity, which is complementary to the other operate functions of protect and constrain. DE (Health) can play a unique role in engagement, given its ability to develop new partnerships through health-related activity. DE (Health) may be an enabler for other engagements or to enable the protect and constrain functions. This will be dependent on delivering improvement in health outcomes. Therefore, the DE (Health) practitioner must be conversant with both the contemporary defence and global health contexts to deliver effective DE (Health) activities. This is an article commissioned for the DE special issue of BMJ Military Health.

Comparison between Defence Healthcare Engagement and humanitarian assistance.

Humanitarian assistance and Defence Healthcare Engagement have traditionally both been taught on the Medical Humanitarian Stabilisation Operations Course. However, the two activities are distinct. This paper outlines the critical differences between them, focusing on their specific purposes, scope, timescales and ethics. Humanitarian assistance will remain a distinct activity with a focus on the relief of suffering, guided by international norms, while Defence Healthcare Engagement will encompass a broader range of activities, less constrained by internationally agreed principles. This presents an opportunity for the Defence Medical Services to directly contribute to projecting UK influence, preventing conflict and building stability. However, it requires the Defence Medical Services to take responsibility for the ethical issues that Defence Healthcare Engagement raises. This paper recommends the development of an ethical framework that reconciles the strategic aims of Defence Healthcare Engagement with maximising patient welfare at the tactical level. This is a paper commissioned as a part of the Humanitarian and Disaster Relief Operations special issue of BMJ Military Health.

Prehospital emergency care in a humanitarian environment: an overview of the ethical considerations.

Recent history has demonstrated that UK Defence personnel can be used, potentially with little notice, in humanitarian disaster zones. The provision of prehospital emergency care (PHEC) in a humanitarian environment requires an innovative approach to overcome the technical challenges of a resource-limited setting. In addition to technical challenges, prehospital practitioners working in a humanitarian environment can expect to be faced with ethically testing situations that they are not familiar with in their usual practice. The organisational and individual ethical decision-making burden can result in significant harms. Therefore, the aim of this paper is to discuss the ethical considerations relevant to providing PHEC during a humanitarian disaster in order that personnel can be more prepared to optimally deliver care. This is a paper commissioned as a part of the Humanitarian and Disaster Relief Operations special issue of BMJ Military Health.