Influential factors in nectar composition and yield in Leptospermum scoparium : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Plant Science, Institute of Agriculture and the Environment, College of Sciences, Massey University, Palmerston North, New Zealand

Material omitted from digital version of thesis: Nickless, E. M., Anderson, C. W. N., Hamilton, G., Stephens, J. M., & Wargent, J. (2016). Soil influences on plant growth, floral density and nectar yield in three cultivars of manuka (Leptospermum scoparium). New Zealand Journal of Botany, 55(2), 100-117. doi:10.1080/0028825X.2016.1247732 ; Nickless, E. M., Holroyd, S. E., Stephens, J. M., Gordon, K. C., & Wargent, J. J. (2014). Analytical FT-Raman spectroscopy to chemotype Leptospermum scoparium and generate predictive models for screening for dihydroxyacetone levels in floral nectar. Journal of Raman Spectroscopy, 45(10), 890-894. doi:10.1002/jrs.4576 ; Nickless, E. M., Holroyd, S. E., Hamilton, G., Gordon, K. C., & Wargent, J. J. (2016). Analytical method development using FTIR-ATR and FT-Raman spectroscopy to assay fructose, sucrose, glucose and dihydroxyacetone, in Leptospermum scoparium nectar. Vibrational Spectroscopy, 84 (2016), 38-43. doi:10.1016/j.vibspec.2016.02.011Leptospermum scoparium (Mānuka) is the plant nectar source for medically bioactive honey, commercially marketed in New Zealand as Unique Mānuka Factor honey (UMF-honey). Methylglyoxal (MGO) is the unique bioactive component of UMF honey with Mānuka nectar containing significant amounts of the carbohydrate dihydroxyacetone (DHA), the chemical precursor for MGO. Anecdotal evidence and recently published data from nectar samples collected from various cultivars in natural sites or botanical gardens has indicated that the DHA and overall composition of L. scoparium nectar varies according to cultivar. The source of this variation is not clearly understood and although there is considerable literature on climatic and genetic influences on nectar composition and yield within various other plant species, there is little published work available on the influence of genetic and environmental factors on the composition and yield of nectar in L. scoparium. Of value to the commercial UMF honey industry in New Zealand is the ability to assess cultivars from breeding programs for the best potential to increase overall UMF honey yield. Predictive modelling of yields is invaluable to the developing honey industry to allow assessment of environmental influences that may affect overall yield along with seasonal influences on nectar production in Mānuka. The research in this thesis establishes the effect of various parameters on overall DHA yield from Mānuka and the beginnings of modelling influencing environmental factors. To determine influences on dihydroxyacetone (DHA) concentration and yield in the nectar of L. scoparium a number of studies were carried out. Methodologies for the collection and analysis of nectar were established. Ten different cultivars of L. scoparium with a range of genetic parentage were studied in controlled glasshouse conditions to assess phenotypic variability in terms of nectar composition and yield as well as plant growth and flowering amongst these cultivars. Significant differences in plant growth and flowering habits were observed amongst the ten cultivars, significant differences in nectar yield and nectar composition with regard to DHA yield were also observed. DHA yields ranged from 2714-7459 mg of DHA/kilogram normalised to 80 oBRIX, with total nectar sugar yields ranging between 0.7 and 4.8 mg amongst the ten cultivars studied. Preliminary research into the effect of temperature, radiation and humidity on nectar composition and yield were also undertaken. Effects of soil composition on these same parameters were researched with a subset of three of the ten cultivars grown on ten different soil types. Plant relative growth rates, dry weights and total plant height were measured throughout a 15 month glasshouse trial. Plant growth, flowering phenology, floral density, nectar yield and DHA composition data was gathered. Soils were analysed for various macronutrient and micronutrient levels and these parameters were modelled against plant data to determine which soil components were influencing plant parameters of interest. Soil type was shown to have no significant effect on DHA concentrations in nectar but results did show that soil type had a significant effect on flowering density amongst the three L. scoparium cultivars studied in the trial. Results from regression analysis of soil chemistry against measured plant parameters indicate that a fertiliser regime has the potential to increase nectar yields due to increased flower numbers. Multivariate analysis using partial least squares regression of soil composition data against plant parameters of value showed that soil components; phosphorus, sulphate, ferric and chloride were commonly shown to influence plant parameters measured. Analytical spectroscopy was investigated as a method to chemotype L. scoparium cultivars and also as a method for quantifying nectar components sucrose, glucose, fructose and DHA. Nectar composition was analysed using high pressure liquid chromatography (HPLC) and compared with fourier transform Raman spectroscopy (FT-Raman) and attenuated total reflectance infrared spectroscopy (ATR-FTIR) analytical spectroscopy methods. FT-Raman spectroscopy was shown to be useful in chemotyping cultivars and in addition proved to be a useful analytical method to predict DHA yield using leaf material from L. scoparium plants from the ten cultivars. FT-Raman and ATR-FTIR proved to be relatively accurate techniques to quantify L. scoparium nectar components DHA, fructose, glucose and sucrose, compared with HPLC methods which use extensive preparation techniques. R-squared values were very good for all nectar components measured excepting the sucrose model at R2 = 0.77. The R2 for the FT-Raman predictions of DHA against HPLC data are very good at 0.85. FTIR prediction data against HPLC data was also good at 0.86 R2. Overall an accurate model is possible for quantifying DHA concentrations in nectar using both FTIR-ATR and FT-Raman spectroscopy. Overall results show that various factors need to be considered when assessing plants for commercial use in the (UMF) Mānuka honey industry within New Zealand. Due to their large impact on overall nectar yield; floral density and plant growth rate parameters are the two key factors of value for commercial assessment of Mānuka cultivars. This research also highlights the importance of assessing not just DHA concentration in deducing cultivar value, but overall nectar yield. These key features must be explored when assessing L. scoparium plants within breeding programs, prior to selection for large-scale field production of high UMF Mānuka honey

The use of simulation to address the acute care skills deficit in pre-registration nursing students: A clinical skills perspective

The increase in patient acuity in primary and secondary settings is continuing, with a corresponding increase in the need for technological competence in these areas. Evidence, however, both nationally and internationally, suggests that these expectations are not being met. This paper offers a review of the literature on acute care, with a specific focus on pre-registration nursing students and the development of acute care skills. Three themes are discussed: factors contributing to the acute care skills deficit, the knowledge and skills required to work in acute care and strategies used to support the acquisition of acute care skills. In response to the review, and based upon the evidence-based solutions identified, the clinical skills team at Bournemouth University designed and developed two teaching sessions, using simulation and role play to support the acquisition of acute care skills in pre-registration students. Student evaluations identify that their knowledge, competence and confidence in this area have increased following the teaching sessions, although caution remains regarding transferability of these skills into the practice environment

Education’s not black and white, it’s vibrant grey

This paper offers a learner’s eye-view of a journey through education, written in an auto-narrative style. Sarah’s story spans from Secondary School to College and through University finishing at the point of Graduation. Revealed in this open and honest account is an insight to coping with home life whilst navigating the trials of the Education system. Sarah makes criticisms of her peers at every level reminding us how competitive some students have to be to get on and achieve success in their education. Critical judgements are also made about Sarah’s School teachers and University lecturers, which are at times as rewarding as they are uncomfortable, but always truthful. Consequently, there is much to learn by both academics and students from this sensitive and vulnerable personal revelation. Sarah’s evidence in turn points to some fundamental questions about the genuine outcomes of the Educational system, e.g. what are we actually teaching young people to be like? And do we like the product in terms of their values, beliefs and motives? A concluding message from Sarah’s perspective is that greater independence in learning, freedom in thinking and equipping people to reason, judge and make decisions in whatever realm, may be defining steps towards becoming educated

Compound C inhibits nonsense-mediated RNA decay independently of AMPK

The nonsense mediated RNA decay (NMD) pathway safeguards the integrity of the transcriptome by targeting mRNAs with premature translation termination codons (PTCs) for degradation. It also regulates gene expression by degrading a large number of non-mutant RNAs (including mRNAs and noncoding RNAs) that bear NMD-inducing features. Consequently, NMD has been shown to influence development, cellular response to stress, and clinical outcome of many genetic diseases. Small molecules that can modulate NMD activity provide critical tools for understanding the mechanism and physiological functions of NMD, and they also offer potential means for treating certain genetic diseases and cancer. Therefore, there is an intense interest in identifying small-molecule NMD inhibitors or enhancers. It was previously reported that both inhibition of NMD and treatment with the AMPK-selective inhibitor Compound C (CC) induce autophagy in human cells, raising the possibility that CC may be capable of inhibiting NMD. Here we show that CC indeed has a NMD-inhibitory activity. Inhibition of NMD by CC is, however, independent of AMPK activity. As a competitive ATP analog, CC does not affect the kinase activity of SMG1, an essential NMD factor and the only known kinase in the NMD pathway. However, CC treatment down-regulates the protein levels of several NMD factors. The induction of autophagy by CC treatment is independent of ATF4, a NMD target that has been shown to promote autophagy in response to NMD inhibition. Our results reveal a new activity of CC as a NMD inhibitor, which has implications for its use in basic research and drug development

Tracer concentration profiles measured in central London as part of the REPARTEE campaign

There have been relatively few tracer experiments carried out that have looked at vertical plume spread in urban areas. In this paper we present results from two tracer (cyclic perfluorocarbon) experiments carried out in 2006 and 2007 in central London centred on the BT Tower as part of the REPARTEE (Regent’s Park and Tower Environmental Experiment) campaign. The height of the tower gives a unique opportunity to study vertical dispersion profiles and transport times in central London. Vertical gradients are contrasted with the relevant Pasquill stability classes. Estimation of lateral advection and vertical mixing times are made and compared with previous measurements. Data are then compared with a simple operational dispersion model and contrasted with data taken in central London as part of the DAPPLE campaign. This correlates dosage with non-dimensionalised distance from source. Such analyses illustrate the feasibility of the use of these empirical correlations over these prescribed distances in central London

Greenhouse gas network design using backward Lagrangian particle dispersion modelling − Part 1: Methodology and Australian test case

This paper describes the generation of optimal atmospheric measurement networks for determining carbon dioxide fluxes over Australia using inverse methods. A Lagrangian particle dispersion model is used in reverse mode together with a Bayesian inverse modelling framework to calculate the relationship between weekly surface fluxes, comprising contributions from the biosphere and fossil fuel combustion, and hourly concentration observations for the Australian continent. Meteorological driving fields are provided by the regional version of the Australian Community Climate and Earth System Simulator (ACCESS) at 12 km resolution at an hourly timescale. Prior uncertainties are derived on a weekly timescale for biosphere fluxes and fossil fuel emissions from high-resolution model runs using the Community Atmosphere Biosphere Land Exchange (CABLE) model and the Fossil Fuel Data Assimilation System (FFDAS) respectively. The influence from outside the modelled domain is investigated, but proves to be negligible for the network design. Existing ground-based measurement stations in Australia are assessed in terms of their ability to constrain local flux estimates from the land. We find that the six stations that are currently operational are already able to reduce the uncertainties on surface flux estimates by about 30%. A candidate list of 59 stations is generated based on logistic constraints and an incremental optimisation scheme is used to extend the network of existing stations. In order to achieve an uncertainty reduction of about 50%, we need to double the number of measurement stations in Australia. Assuming equal data uncertainties for all sites, new stations would be mainly located in the northern and eastern part of the continent

Investigation of mechanistic deterioration modeling for bridge design and management

2017 Spring.Includes bibliographical references.The ongoing deterioration of highway bridges in Colorado dictates that an effective method for allocating limited management resources be developed. In order to predict bridge deterioration in advance, mechanistic models which analyze the physical processes causing deterioration are capable of supplementing purely statistical models and addressing limitations associated with bridge inspection data and statistical methods. A review of existing analytical models in the literature was conducted. Due to its prevalence throughout the state of Colorado and frequent need for repair, corrosion-induced cracking of reinforced concrete (RC) decks was selected as the mode of deterioration for further study. A mechanistic model was developed to predict corrosion and concrete cracking as a function of material and environmental inputs. The model was modified to include the effects of epoxy-coated rebar, waterproofing membranes, asphalt overlays, joint deterioration, and deck maintenance. Probabilistic inputs were applied to simulate inherent randomness associated with deterioration. Model results showed that mechanistic models may be able to address limitations of statistical models and provide a more accurate and precise prediction of bridge degradation in advance. Preventative maintenance may provide longer bridge deck service life with fewer total maintenance actions than current methods. However, experimental study of specific deterioration processes and additional data collection are needed to validate model predictions. Maintenance histories of existing bridges are necessary to predicting bridge deterioration and improving bridge design and management in the future

RAG-induced DNA double-strand breaks signal through Pim2 to promote pre-B cell survival and limit proliferation

Interleukin 7 (IL-7) promotes pre–B cell survival and proliferation by activating the Pim1 and Akt kinases. These signals must be attenuated to induce G1 cell cycle arrest and expression of the RAG endonuclease, which are both required for IgL chain gene rearrangement. As lost IL-7 signals would limit pre–B cell survival, how cells survive during IgL chain gene rearrangement remains unclear. We show that RAG-induced DNA double-strand breaks (DSBs) generated during IgL chain gene assembly paradoxically promote pre–B cell survival. This occurs through the ATM-dependent induction of Pim2 kinase expression. Similar to Pim1, Pim2 phosphorylates BAD, which antagonizes the pro-apoptotic function of BAX. However, unlike IL-7 induction of Pim1, RAG DSB-mediated induction of Pim2 does not drive proliferation. Rather, Pim2 has antiproliferative functions that prevent the transit of pre–B cells harboring RAG DSBs from G1 into S phase, where these DNA breaks could be aberrantly repaired. Thus, signals from IL-7 and RAG DSBs activate distinct Pim kinase family members that have context-dependent activities in regulating pre–B cell proliferation and survival