Rubredoxin Protein Scaffolds Sourced from Diverse Environmental Niches as an Artificial Hydrogenase Platform.

Nickel-substituted rubredoxin (NiRd) from Desulfovibrio desulfuricans has previously been shown to act as both a structural and functional mimic of the [NiFe] hydrogenase. However, improvements both in turnover frequency and overpotential are needed to rival the native [NiFe] hydrogenase enzymes. Characterization of a library of NiRd mutants with variations in the secondary coordination sphere suggested that protein dynamics played a substantial role in modulating activity. In this work, rubredoxin scaffolds were selected from diverse organisms to study the effects of distal sequence variation on catalytic activity. It was found that though electrochemical catalytic activity was only slightly impacted across the series, the Rd sequence from a psychrophilic organism exhibited substantially higher levels of solution-phase hydrogen production. Additionally, Eyring analyses suggest that catalytic activation properties relate to the growth temperature of the parent organism, implying that the general correlation between the parent organism environment and catalytic activity often seen in naturally occurring enzymes may also be observed in artificial enzymes. Selecting protein scaffolds from hosts that inhabit diverse environments, particularly low-temperature environments, represents an alternative approach for engineering artificial metalloenzymes.

Vaccination terminology: A revised glossary of key terms including lay person's definitions.

WHAT IS KNOWN AND OBJECTIVE: There has been a paucity of vaccine and vaccine-related definitions within the scientific and medical peer-reviewed literature, particularly with the arrival of COVID-19. Therefore, it was the aim of this commentary to collate definitions to 44 vaccine- and vaccinology-related key terms, from four international and respected sources of information (where available), including (i) the World Health Organisation (WHO), (ii) the US Centers for Disease Control and Prevention (CDC), (iii) The Department of Health, Government of Australia and (iv) the European Union. In addition, it was a further aim to develop a lay person's definition to each of these 44 key terms, to act as a published and citeable reference point for pharmacists and other healthcare professionals, when communicating with patients and other public-facing stakeholders. COMMENT: Definitions are important in health care in order to (i) provide concise insight on a specific topic, (ii) provide a common understanding and (iii) set reference points to allow the adoption of a standard uniform approach. WHAT IS NEW AND CONCLUSION: The collation of definitions of key vaccine terms was compiled from four respected sources of information. A glossary of 44 key terms was produced to help pharmacists and other healthcare professionals explain such terms professionally, as well as to patient stakeholders in lay person's vocabulary. These lay definitions had superior readability metrics than definitions from any of the four professional sources, indicating their suitability for engagement with patient-facing stakeholders. Understanding the barriers to vaccine uptake is crucial for health professionals and policymakers to achieve improved uptake rates. This commentary has aimed at adding value to healthcare professionals and patients, by providing an up-to-date glossary of several professional definitions, from respected sources, as well as an accompanying lay definition to support the healthcare professional-patient communicative interface. Vaccines have become an important preventative tool, particularly in the context of the COVID-19 pandemic, to help mitigate disease severity and to help control the pandemic locally, nationally and internationally. Accessible and robust definitions help inform the dialogue to achieve this goal and the avoidance of obscurum per obscurius.

In Vivo Assembly of a Genetically Encoded Artificial Metalloenzyme for Hydrogen Production.

The genetic encoding of artificial enzymes represents a substantial advantage relative to traditional molecular catalyst optimization, as laboratory-based directed evolution coupled with high-throughput screening methods can provide rapid development and functional characterization of enzyme libraries. However, these techniques have been of limited utility in the field of artificial metalloenzymes due to the need for in vitro cofactor metalation. Here, we report the development of methodology for in vivo production of nickel-substituted rubredoxin, an artificial metalloenzyme that is a structural, functional, and mechanistic mimic of the [NiFe] hydrogenases. Direct voltammetry on cell lysate establishes precedent for the development of an electrochemical screen. This technique will be broadly applicable to the in vivo generation of artificial metalloenzymes that require a non-native metal cofactor, offering a route for rapid enzyme optimization and setting the stage for integration of artificial metalloenzymes into biochemical pathways within diverse hosts.

Meningococcal ACWY vaccine uptake and awareness among student freshers enrolled at Northern Ireland universities.

A new MenACWY conjugate meningococcal vaccination programme was introduced in Northern Ireland (NI) in August 2015, for 13-18 year olds, as well as for first-time university entrants up to 25 years. This reflected the response made by Public Health England, due to the recent rapid increase of meningococcal group W (MenW) disease and on advice of the UK Joint Committee on Vaccination and Immunisation (JCVI). The aims of this study were to evaluate (i) the uptake of the MenACWY vaccine among first-time university students, (ii) vaccine and meningitis awareness, (iii) optimal communication modalities via a multidisciplinary team (MDT) model and (iv) current international vaccination policies relating to non-UK students. A survey was completed by 1210 students, 868 first-time freshers and 342 non-freshers, from healthcare-related, non-healthcare-related and engineering/computing faculties. The survey included an anonymous questionnaire and consented students were aged 17-50 years with a 2:3 ratio of male:female. Vaccine uptake amongst 18-year-old students was 90.7% and 87.3% in female and male cohorts, respectively, falling to 72.1% and 67.7% (19-year cohort) and 32.7% and 39.6% (20- to 25-year cohort) in males and females, respectively. Students reported that posters, clinics and talks were the preferred methods of communication and not social media. There was general lack of awareness of the signs/symptoms of meningitis and approximately 30% of students falsely believed that administration of the MenACWY vaccine excluded the risk of contracting meningitis. Overall, there was a successful vaccination campaign; however, there was a lack of meningitis awareness. Due to differing international meningococcal vaccination schedules, international students enrolling at UK universities need to be informed about current UK policies. For the successful introduction of any vaccination programme amongst university students, it is fundamental that a MDT is established to inform and deliver such a programme in an efficient and timely manner.

Molecular identification of airborne bacteria associated with aerial spraying of bovine slurry waste employing 16S rRNA gene PCR and gene sequencing techniques.

Polymerase chain reaction amplification of the universal 16S ribosomal RNA (rRNA) gene was performed on a collection of 38 bacterial isolates, originating from air sampled immediately adjacent to the agricultural spreading of bovine slurry. A total of 16 bacterial genera were identified including both Gram-positive and Gram-negative genera. Gram-positive organisms accounted for 34/38 (89.5%) of total bacterial numbers consisting of 12 genera and included Staphylococcus (most common genus isolated), Arthrobacter (2nd most common genus isolated), Brachybacterium, Exiguobacterium, Lactococcus, Microbacterium and Sporosarcina (next most common genera isolated) and finally, Bacillus, Brevibacterium, Frigoribacterium, Mycoplana and Pseudoclavibacter. Gram-negative organisms accounted for only 4/38 (10.5%) bacterial isolates and included the following genera, Brevundimonas, Lysobacter, Psychrobacter and Rhizobium. No gastrointestinal pathogens were detected. Although this study demonstrated a high diversity of the microorganisms present, only a few have been shown to be opportunistically pathogenic to humans and none of these organisms described have been described previously as having an inhalational route of infection and therefore we do not believe that the species of organisms identified pose a significant health and safety threat for immunocompetant individuals.

Massive gastrointestinal haemorrhage due to gastritis cystica profunda.

INTRODUCTION: Gastritis Cystica Profunda is a well recognized entity which may occur several years after previous gastric surgery. This is a premalignant condition and may lead on to carcinoma of the stomach. CASE PRESENTATION: We report a case of a 50-year-old man with epigastric pain and haematemesis. 28 years ago he had undergone partial gastrectomy and gastroenterostomy for benign gastric ulcer. An Upper gastrointestinal endoscopy showed a possible bleeding vessel on the anterior wall lesser curve of the stomach. The lesion was injected with adrenaline 1 in 100,000. In spite of the intervention he continued to have haemetemesis with significant haemodynamic impairment. At exploratory laparotomy, an oedematous ridge on the posterior wall with a bleeding point on the posterior gastric wall. Histology showed features consistent with gastritis cystica profunda. He made an excellent post-operative recovery. CONCLUSION: We suggest that patients who are diagnosed with gastritis cystica profunda should be regularly followed up as this is a premalignant condition.

Methods for identifying surgical wound infection after discharge from hospital: a systematic review.

BACKGROUND: Wound infections are a common complication of surgery that add significantly to the morbidity of patients and costs of treatment. The global trend towards reducing length of hospital stay post-surgery and the increase in day case surgery means that surgical site infections (SSI) will increasingly occur after hospital discharge. Surveillance of SSIs is important because rates of SSI are viewed as a measure of hospital performance, however accurate detection of SSIs post-hospital discharge is not straightforward. METHODS: We conducted a systematic review of methods of post discharge surveillance for surgical wound infection and undertook a national audit of methods of post-discharge surveillance for surgical site infection currently used within United Kingdom NHS Trusts. RESULTS: Seven reports of six comparative studies which examined the validity of post-discharge surveillance methods were located; these involved different comparisons and some had methodological limitations, making it difficult to identify an optimal method. Several studies evaluated automated screening of electronic records and found this to be a useful strategy for the identification of SSIs that occurred post discharge. The audit identified a wide range of relevant post-discharge surveillance programmes in England, Scotland and Wales and Northern Ireland; however, these programmes used varying approaches for which there is little supporting evidence of validity and/or reliability. CONCLUSION: In order to establish robust methods of surveillance for those surgical site infections that occur post discharge, there is a need to develop a method of case ascertainment that is valid and reliable post discharge. Existing research has not identified a valid and reliable method. A standardised definition of wound infection (e.g. that of the Centres for Disease Control) should be used as a basis for developing a feasible, valid and reliable approach to defining post discharge SSI. At a local level, the method used to ascertain post discharge SSI will depend upon the purpose of the surveillance, the nature of available routine data and the resources available.

HPLC-SPE-NMR in pharmaceutical development: capabilities and applications.

High-performance liquid chromatography-solid phase extraction-NMR spectroscopy (HPLC-SPE-NMR) has recently become commercially available and has been evaluated with regard to its applicability in a pharmaceutical environment. The addition of an automated SPE unit to an HPLC-NMR system for peak trapping results in an improved NMR signal-to-noise ratio (S/N) and also has other practical advantages. The trapping efficiency is shown to depend on compound polarity and is highest for compounds eluting late on reversed-phase HPLC systems. Multiple peak trapping further increases the S/N, again with the best results for less polar compounds. For polar compounds, multiple peak trapping resulted in no S/N gain as the amount of material retained on the SPE cartridge was equivalent to that from a single injection. When compared with conventional HPLC-NMR, a S/N gain of up to five-fold could be achieved for some compounds in a single trapping step. A major advantage of the technique is the independence of the chromatographic step from the NMR step, resulting in greater versatility than conventional HPLC-NMR in the HPLC solvents and NMR solvents that can be used. Practical applications from both drug metabolite and drug impurity identification are presented.