Chemical Analysis and Elucidation of Anthraquinone and Flavonoid Type Compounds with Applications to Historical Artefacts and Sustainability

This thesis describes the effects of different solvents have on the extraction profile of natural dyes from dye plants: madder, weld, golden rod, and chamomile. HPLC has been used to build a fingerprint of each dye plant profile and thus used to compare to the profiles of back extraction from textiles for natural dye identification in historical artefacts. The use of solid phase extraction is compared to extraction methods with no purification step which is favourable for anthraquinone dyes but results in a major loss of glycosidic compounds when using yellow dyestuffs. Supported by 1H and 13C NMR data, the conclusive X-ray crystal structure of the natural dye ruberythric acid is presented which has never been achieved prior to this research. In a collaboration with food science two of the main components of chamomile are fully characterised by 1D and 2D, 1H and 13C NMR. These compounds are usually referred to as ferulic acid derivatives in the literature but their actual structure is reported herein. The thesis also discusses the relative dye uptake of anthraquinone compounds onto wool textiles which were measured by HPLC. Sorption isotherms for the main anthraquinones in madder; ruberythric acid, pseudopurpurin and alizarin are compared for more in-depth understanding on the method of adsorption of these compounds. Herein the glycosidic compounds in madder are shown to have a higher adsorption capacity than the aglycons. Ruberythric acid is shown to follow a Tempkin isotherm with the highest degree of correlation but both alizarin and ruberythric acid show good fitting with the Freundlich isotherm also. Pseudopurpurin was shown to follow a Freundlich isotherm with the highest degree of linearity but did also show some fitting to the Langmuir isotherm. The isotherms allow data to be collected on the energy of adsorption and draw conclusions on the effect the functional groups have on the dyeing capability which is studied herein for the first time on individual anthraquinone components. iii Studies were carried out on the acid-sensitive colorants present in madder which are degraded in the textile back extraction process. Anthraquinone aglycons alizarin and purpurin are usually identified in analysis following harsh back extraction methods, such as those using solvent mixtures with concentrated hydrochloric acid at high temperatures. Herein, a softer novel extraction method involving aqueous glucose solution was developed and compared to other back extraction techniques on wool dyed with root extract from different varieties of Rubia tinctorum. A study into the breakdown compounds of the aglycon; lucidin under acidic conditions used for traditional back extractions was also undertaken. Here it is observed that lucidin is converted into xanthopurpurin in a retro aldol like mechanism. This report discusses some of the issues raised by using these harsh back extraction methods and the problems faced in using them to analyse historic artefacts

Translating research into practice: Evaluation of an e-learning resource for health care professionals to provide nutrition advice and support for cancer survivors

Background Nurses and other allied health professionals are in a key position to provide appropriate and consistent advice on nutritional issues to support cancer survivors. However gaps in their nutrition knowledge and education warrant the need for enhanced learning as part of their Continued Professional Development (CPD). In the UK there are currently no formally recognised nutrition education programmes. Therefore e-learning offers a solution to provide flexible learning to target this need. Aims This study aimed to develop and evaluate the efficacy of a freely available, internet-based learning resource, for nurses and allied health professionals who provide nutrition, diet and lifestyle advice for cancer survivors. It sought to explore the attitudes and conceptions of the resource and current knowledge base of those involved in the care pathway for cancer survivors. Method The design and development of the e-learning resource were informed by the best available research and policy evidence and in a format to facilitate on-line learning. A robust evaluation strategy incorporated focus groups and telephone interviews to gain in depth insights into the experiences of using the resource. Results Themes included ‘Plugging a Gap’ which shows an improved knowledge base for nutrition. Information was ‘All in One Place’ showing that the resource was valued as being within a ‘trusted’ organisation. ‘Everyone Benefits’ illustrates how learners felt that the resource provided them with an evidence base, whilst the ‘Current and Live’ theme captured how professionals felt about the information being up-to-date. Conclusions The project has shown the benefits of interprofessional working to develop an e-learning resource for Health Care Professionals to support cancer survivors in following healthier lifestyles. Positive attitudes and potential improvements in the knowledge base and changes for professional practice were demonstrated. Further research is required to gauge sustained impact in the work environment by embedding the resource as part of CPD

Lipoprotein biosynthesis as a target for anti-Wolbachia treatment of filarial nematodes

<p>Abstract</p> <p>Background</p> <p>Lymphatic filariasis and onchocerciasis are debilitating diseases caused by filarial nematodes. Disease pathogenesis is induced by inflammatory responses following the death of the parasite. <it>Wolbachia </it>endosymbionts of filariae are potent inducers of innate and adaptive inflammation and bacterial lipoproteins have been identified as the ligands that bind toll-like receptors (TLR) 2 and TLR6. Lipoproteins are important structural and functional components of bacteria and therefore enzymes involved in <it>Wolbachia </it>lipoprotein biosynthesis are potential chemotherapeutic targets.</p> <p>Results</p> <p>Globomycin, a signal peptidase II (LspA) inhibitor, has activity against Gram-negative bacteria and a putative <it>lspA </it>gene has been identified from the <it>Wolbachia </it>genome of <it>Brugia malayi </it>(<it>w</it>Bm). The amino acids required for function are strictly conserved and functionality was verified by complementation tests in a temperature-sensitive <it>Escherichia coli lspA </it>mutant. Also, transformation of wild type <it>E. coli </it>with <it>Wolbachia lspA </it>conferred significant globomycin resistance. A cell-based screen has been developed utilizing a <it>Wolbachia</it>-containing <it>Aedes albopictus </it>cell line to assay novel compounds active against <it>Wolbachia</it>. Globomycin was screened using this assay, which resulted in a dose-dependent reduction in <it>Wolbachia </it>load. Furthermore, globomycin was also effective in reducing the motility and viability of adult <it>B. malayi in vitro</it>.</p> <p>Conclusions</p> <p>These studies validate lipoprotein biosynthesis as a target in an organism for which no genetic tools are available. Further studies to evaluate drugs targeting this pathway are underway as part of the A-WOL drug discovery and development program.</p

Mental health at the COVID-19 frontline : an assessment of distress, fear, and coping among staff and attendees at screening clinics of rural/regional settings of Victoria, Australia

Purpose: Research examining psychological well-being associated with COVID-19 in rural/regional Australia is limited. This study aimed to assess the extent of psychological distress, fear of COVID-19, and coping strategies among the attendees in COVID-19 screening clinics at 2 rural Victorian settings. Methods: A cross-sectional study was conducted during July 2020 to February 2021 inclusive. Participants were invited to fill in an online questionnaire. Kessler Psychological Distress Scale (K-10), Fear of COVID-19 Scale, and Brief Resilient Coping Scale were used to assess psychological distress, fear of COVID-19, and coping, respectively. Findings: Among 702 total participants, 69% were females and mean age (±SD) was 49 (±15.8) years. One in 5 participants (156, 22%) experienced high to very high psychological distress, 1 in 10 (72, 10%) experienced high fear, and more than half (397, 57%) had medium to high resilient coping. Participants with mental health issues had higher distress (AOR 10.4, 95% CI: 6.25-17.2) and fear (2.56, 1.41-4.66). Higher distress was also associated with having comorbidities, increased smoking (5.71, 1.04-31.4), and alcohol drinking (2.03, 1.21-3.40). Higher fear was associated with negative financial impact, drinking alcohol (2.15, 1.06-4.37), and increased alcohol drinking. Medium to high resilient coping was associated with being ≥60 years old (1.84, 1.04-3.24) and completing Bachelor and above levels of education. Conclusion: People who had pre-existing mental health issues, comorbidities, smoked, and consumed alcohol were identified as high-risk groups for poorer psychological well-being in rural/regional Victoria. Specific interventions to support the mental well-being of these vulnerable populations, along with engaging health care providers, should be considered. © 2021 The Authors. The Journal of Rural Health published by Wiley Periodicals LLC on behalf of National Rural Health Association

Assessing inhaler techniques of asthma patients using aerosol inhalation monitors (aim): a cross-sectional study

A high percentage of asthma patients have symptoms that are not well controlled, despite effective drugs being available. One potential reason for this may be that poor inhaler technique limits the dose delivered to the lungs, thereby reducing the therapeutic efficacy. The aim of this study was to assess the prevalence of poor inhaler technique in an asthma patient population and to probe the impact of various demographic parameters on technique quality. This study was conducted at community pharmacies across Wales, UK. Patients diagnosed with asthma and 12 years or older were invited to participate. An aerosol inhalation monitor (AIM, Vitalograph®) was used to measure the quality of patient inhaler technique. A total of 295 AIM assessments were carried out. There were significant differences in the quality of inhaler technique across the different inhaler types (p < 0.001, Chi squared). The best technique was associated with dry-powder inhalers (DPI devices, 58% of 72 having good technique), compared with pressurized metered-dose inhalers (pMDI) or pMDIs with a spacer device (18% of 174 and 47% of 49 AIM assessments, respectively). There were some significant associations between gender, age, and quality of inhaler technique, as determined with adjusted odds ratios. It seems that the majority of asthmatic patients were not using their inhalers appropriately. We recommend that healthcare professionals place more emphasis on assessing and correcting inhaler technique, as poor inhaler technique might be responsible for the observed lack of symptom control in the asthma patient population

Absence of Wolbachia endobacteria in the non-filariid nematodes Angiostrongylus cantonensis and A. costaricensis

The majority of filarial nematodes harbour Wolbachia endobacteria, including the major pathogenic species in humans, Onchocerca volvulus, Brugia malayi and Wuchereria bancrofti. These obligate endosymbionts have never been demonstrated unequivocally in any non-filariid nematode. However, a recent report described the detection by PCR of Wolbachia in the metastrongylid nematode, Angiostrongylus cantonensis (rat lungworm), a leading cause of eosinophilic meningitis in humans. To address the intriguing possibility of Wolbachia infection in nematode species distinct from the Family Onchocercidae, we used both PCR and immunohistochemistry to screen samples of A. cantonensis and A. costaricensis for the presence of this endosymbiont. We were unable to detect Wolbachia in either species using these methodologies. In addition, bioinformatic and phylogenetic analyses of the Wolbachia gene sequences reported previously from A. cantonensis indicate that they most likely result from contamination with DNA from arthropods and filarial nematodes. This study demonstrates the need for caution in relying solely on PCR for identification of new endosymbiont strains from invertebrate DNA samples

Is bureaucracy being busted in research ethics and governance for health services research in the UK?:Experiences and perspectives reported by stakeholders through an online survey

Acknowledgements: We would like to thank the Board of Trustees of HSRUK for support and advice throughout study. Thank you to all respondents to the survey for contributing their data and views. The HRA and NIHR are also warmly acknowledged for their continued support throughout the study.Peer reviewedPublisher PD

The Diabetes Remission Clinical Trial (DiRECT): protocol for a cluster randomised trial

Background: Despite improving evidence-based practice following clinical guidelines to optimise drug therapy, Type 2 diabetes (T2DM) still exerts a devastating toll from vascular complications and premature death. Biochemical remission of T2DM has been demonstrated with weight loss around 15kg following bariatric surgery and in several small studies of non-surgical energy-restriction treatments. The non-surgical Counterweight-Plus programme, running in Primary Care where obesity and T2DM are routinely managed, produces &gt;15 kg weight loss in 33 % of all enrolled patients. The Diabetes UK-funded Counterpoint study suggested that this should be sufficient to reverse T2DM by removing ectopic fat in liver and pancreas, restoring first-phase insulin secretion. The Diabetes Remission Clinical Trial (DiRECT) was designed to determine whether a structured, intensive, weight management programme, delivered in a routine Primary Care setting, is a viable treatment for achieving durable normoglycaemia. Other aims are to understand the mechanistic basis of remission and to identify psychological predictors of response. Methods/Design: Cluster-randomised design with GP practice as the unit of randomisation: 280 participants from around 30 practices in Scotland and England will be allocated either to continue usual guideline-based care or to add the Counterweight-Plus weight management programme, which includes primary care nurse or dietitian delivery of 12-20weeks low calorie diet replacement, food reintroduction, and long-term weight loss maintenance. Main inclusion criteria: men and women aged 20-65years, all ethnicities, T2DM 0-6years duration, BMI 27-45 kg/m2. Tyneside participants will undergo Magnetic Resonance (MR) studies of pancreatic and hepatic fat, and metabolic studies to determine mechanisms underlying T2DM remission. Co-primary endpoints: weight reduction ≥ 15 kg and HbA1c &lt;48 mmol/mol at one year. Further follow-up at 2 years. Discussion: This study will establish whether a structured weight management programme, delivered in Primary Care by practice nurses or dietitians, is a viable treatment to achieve T2DM remission. Results, available from 2018 onwards, will inform future service strategy

Characterization of a Novel Filarial Serine Protease Inhibitor, Ov-SPI-1, from Onchocerca volvulus, with Potential Multifunctional Roles During Development of the Parasite

A novel filarial serine protease inhibitor (SPI) from the human parasitic nematode Onchocerca volvulus, Ov-SPI-1, was identified through the analysis of a molting third-stage larvae expressed sequence tag dataset. Subsequent analysis of the expressed sequence tag datasets of O. volvulus and other filariae identified four other members of this family. These proteins are related to the low molecular weight SPIs originally isolated from Ascaris suumwhere they are believed to protect the parasite from host intestinal proteases. The two Ov-spi transcripts are up-regulated in the molt- ing larvae and adult stages of the development of the parasite. Recombinant Ov-SPI-1 is an active inhibitor of serine proteases, specifically elastase, chymotrypsin, and cathepsin G. Immunolocal- ization of the Ov-SPI proteins demonstrates that the endogenous proteins are localized to the basal layer of the cuticle of third-stage, molting third-stage, and fourth-stage larvae, the body channels and multivesicular bodies of third-stage larvae and the processed mate- rial found between the two cuticles during molting. In O. volvulusadult worms the Ov-SPI proteins are localized to the sperm and to eggshells surrounding the developing embryos. RNA interference targeting the Ov-spi genes resulted in the specific knockdown of the transcript levels of both Ov-spi-1 and Ov-spi-2, a loss of native pro- teins, and a significant reduction in both molting and viability of third-stage larvae. We suggest the Ov-SPI proteins play a vital role in nematode molting by controlling the activity of an endogenous serine protease(s). The localization data in adults also indicate that these inhibitors may be involved in other processes such as embryogenesis and spermatogenesis