The origin and maintenance of a new 'Senecio' hybrid in York, England

The principal aim of the research presented in this thesis was to use a broad range of analytical techniques to investigate the origin and maintenance of a newly arisen Senecio hybrid, referred to as York radiate groundsel, in York, England. The results of morphological and molecular analyses, showed that York radiate groundsel is a hybrid product between Senecio vulgaris var. vulgaris L. and S. squalidus L, and is distinct from the stabilized introgressant, S. vulgaris var. hibernicus Syme. These findings also indicated that York radiate groundsel is unlikely to be a first generation hybrid, but has probably undergone limited backcrossing to S. vulgaris with current populations of the plant derived from a single origin. Tetraploid hybrid progeny between S. squalidus and S. vulgaris were synthesized artificially and some were partially fertile and similar in morphology to York radiate groundsel. It was argued that unreduced S. squalidus gametes are moer likely to have been involved in the origin of the new hybrid. Postzygotic breeding barriers were shown to exist between York radiate groundsel and both of its parents. Backcrosses to S. squalidus were triploid and highly sterile, while backcrosses to S. vulgaris were tetraploid, but exhibited a significant reduction in seed and pollen fertility in the subsequent generation. Although flowering time of York radiate groundsel and S. vulgaris differed substantially, an examination of capitula number in relation to flowering time, revealed that the flowering time difference was probably not selected as a prezygotic isolating mechanism. The results of a field experiment that measured outcrossing rates, suggested that York radiate groundsel may be more attractive to pollinators than either variety of S. vulgaris, which would in turn, lead to its effective ethological reproductive isolation from these taxa. It was also shown that characters that could promote pollinator attraction in the new hybrid, have probably been inherited from S. squalidus and have remained associated with the ray floret locus in York radiate groundsel due to gene linkage. The effects of inbreeding on fitness were found to be negligible in York radiate groundsel and this, together with the fact that it exhibited some novel ecological characteristics, is discussed in regard to its continued maintenance in the wild. The distinctive morphological and molecular phenotype of York radiate groundsel and the fact that it has achieved a level of reproductive isolation from both of its parents, should favour its recognition as a new species

Acute Inflammatory Profiles Differ with Sex and Age After Spinal Cord Injury

Background Sex and age are emerging as influential variables that affect spinal cord injury (SCI) recovery. Despite a changing demographic towards older age at the time of SCI, the effects of sex or age on inflammation remain to be elucidated. This study determined the sex- and age-dependency of the innate immune response acutely after SCI. Methods Male and female mice of ages 4- and 14-month-old received T9 contusion SCI and the proportion of microglia, monocyte-derived macrophages (MDM), and neutrophils surrounding the lesion were determined at 3- and 7-day post-injury (DPI) using flow cytometry. Cell counts of microglia and MDMs were obtained using immunohistochemistry to verify flow cytometry results at 3-DPI. Microglia and MDMs were separately isolated using fluorescence-activated cell sorting (FACS) at 3-day post-injury (DPI) to assess RNA expression of 27 genes associated with activation, redox, and debris metabolism/clearance. Results Flow cytometry revealed that being female and older at the time of injury significantly increased MDMs relative to other phagocytes, specifically increasing the ratio of MDMs to microglia at 3-DPI. Cell counts using immunohistochemistry revealed that male mice have more total microglia within SCI lesions that can account for a lower MDM/microglia ratio. With NanoString analyses of 27 genes, only 1 was differentially expressed between sexes in MDMs; specifically, complement protein C1qa was increased in males. No genes were affected by age in MDMs. Only 2 genes were differentially regulated in microglia between sexes after controlling for false discovery rate, specifically CYBB (NOX2) as a reactive oxygen species (ROS)-associated marker as well as MRC1 (CD206), a gene associated with reparative phenotypes. Both genes were increased in female microglia. No microglial genes were differentially regulated between ages. Differences between microglia and MDMs were found in 26 of 27 genes analyzed, all expressed higher in MDMs with three exceptions. Specifically, C1qa, cPLA2, and CD86 were expressed higher in microglia. Conclusions These findings indicate that inflammatory responses to SCI are sex-dependent at both the level of cellular recruitment and gene expression

Kinetics of Asymmetric Transfer Hydrogenation, Catalyst Deactivation, and Inhibition with Noyori Complexes As Revealed by Real-Time High-Resolution FlowNMR Spectroscopy

Catalytic hydrogen transfer from basic isopropyl alcohol to aryl ketones mediated by [(arene)­(TsDPEN)­RuCl] complexes has been investigated by operando ¹H NMR spectroscopy using a recirculating flow setup. Selective excitation pulse sequences allowed fast and quantitative monitoring of the key [(mesitylene)­(TsDPEN)­RuH] intermediate during catalysis, which is shown to interact with both substrates by polarization transfer experiments. Comparison of reaction profiles with catalyst speciation traces in conjunction with reaction progress kinetic analysis using variable time normalization and kinetic modeling showed the existence of two independent catalyst deactivation/inhibition pathways: whereas excess base exerted a competitive inhibition effect on the unsaturated catalyst intermediate, the active hydride suffered from an inherent first-order decay that is not evident in early stages of the reaction where turnover is fast. Isotopic labeling revealed arene loss to be the entry point into deactivation pathways to Ru nanoparticles via hydride-bridged intermediates

Testing umbrella species and food-web properties of large carnivores in the Rocky Mountains

Despite criticisms, the umbrella species concept remains a fundamental conservation tool for protecting biodiversity in the face of global change, yet it is rarely tested. Food web theory provides a tool to test both umbrella-species' suitability and their ecological function, which we investigate in a large-mammal food web. Using data from 698 camera trap locations in the Canadian Rockies, we develop hierarchical occupancy models to predict the co-occurrence of 16 large mammal species. We draw upon previous diet studies in the Canadian Rockies to describe the meta food-web (meta-web) for these species. Next, we filtered the meta-web using predicted occupancy to estimate realized food webs at each camera location. We tested the umbrella species concept using predicted occupancy across all 698 camera sites. We then tested for carnivore effects using realized food webs on 5 food-web properties: species richness, links, connectance, nestedness and modularity using generalized linear models while accounting for landscape covariates known to affect food web dynamics. Our multispecies occupancy models reflected factors previously demonstrated to affect large mammal occurrence. Our results also demonstrated that grizzly bear (Ursus horribilis), a generalist carnivore, was the best umbrella carivore species, and explained species richness the best. When considering food web properties, however, wolves (Canis lupus) and cougars (Felis concolor) served as better umbrellas that also captured food web properties such as connectance, links and nestedness that better reflect ecological interactions. Our results support the role of large carnivores as umbrella and ecologically interactive species in conservation planning

De novo assembly of red clover transcriptome based on RNA-Seq data provides insight into drought response, gene discovery and marker identification

Background Red clover (Trifolium pratense L.) is a versatile forage crop legume, which can tolerate a variety of soils and is suitable for silage production for winter feed and for grazing. It is one of the most important forage legumes in temperate livestock agriculture. Its beneficial attributes include ability to fix nitrogen, improve soil and provide protein rich animal feed. It is however, a short-lived perennial providing good biomass yield for two or three years. Improved persistency is thus a major breeding target. Better water-stress tolerance is one of the key factors influencing persistency, but little is known about how red clover tolerates water stress. Results Plants from a full sib mapping family were used in a drought experiment, in which the growth rate and relative water content (RWC) identified two pools of ten plants contrasting in their tolerance to drought. Key metabolites were measured and RNA-Seq analysis was carried out on four bulked samples: the two pools sampled before and after drought. Massively parallel sequencing was used to analyse the bulked RNA samples. A de novo transcriptome reconstruction based on the RNA-Seq data was made, resulting in 45181 contigs, representing ‘transcript tags’. These transcript tags were annotated with gene ontology (GO) terms. One of the most striking results from the expression analysis was that the drought sensitive plants were characterised by having approximately twice the number of differentially expressed transcript tags than the tolerant plants after drought. This difference was evident in most of the major GO terms. Before onset of drought the sensitive plants overexpressed a number of genes annotated as senescence-related. Furthermore, the concentration of three metabolites, particularly pinitol, but also proline and malate increased in leaves after drought stress. Conclusions This de novo assembly of a red clover transcriptome from leaf material of droughted and non-droughted plants provides a rich source for gene identification, single nucleotide polymorphisms (SNP) and short sequence repeats (SSR). Comparison of gene expression levels between pools and treatments identified candidate genes for further analysis of the genetic basis of drought tolerance in red clover

Hydrophosphination of unactivated alkenes and alkynes using iron(II):catalysis and mechanistic insight

The catalytic addition of phosphines to alkenes and alkynes is a very attractive process that offers access to phosphines in a 100% atom-economic reaction using readily available and inexpensive materials. The products are potentially useful ligands and organocatalysts. Herein, we report the first example of intramolecular hydrophosphination of a series of nonactivated phosphinoalkenes and phosphinoalkynes with a simple iron β-diketiminate complex. Kinetic studies suggest that this transformation is first-order with respect to both the phosphine and the catalyst. A mechanistic interpretation of the iron-catalyzed hydrophosphination is presented, supported by the experimental evidence collected

Contrasting genetic association of IL2RA with SLE and ANCA-associated vasculitis.

BACKGROUND: Autoimmune diseases are complex and have genetic and environmental susceptibility factors. The objective was to test the genetic association of systemic lupus erythematosus (SLE) and anti-neutrophil cytoplasmic antibody (ANCA) - associated systemic vasculitis (AAV) with SNPs in the IL2RA region and to correlate genotype with serum levels of IL-2RA. METHODS: Using a cohort of over 700 AAV patients, two SLE case-control studies and an SLE trio collection (totalling over 1000 SLE patients), and a TaqMan genotyping approach, we tested 3 SNPs in the IL2RA locus, rs11594656, rs2104286 & rs41295061, each with a prior association with autoimmune disease; rs11594656 and rs41295061 with type 1 diabetes (T1D) and rs2104286 with multiple sclerosis (MS) and T1D. RESULTS: We show that SLE is associated with rs11594656 (P = 3.87 x 10-7) and there is some evidence of association of rs41295061 with AAV (P = 0.0122), which both have prior association with T1D. rs2104286, an MS and T1D - associated SNP in the IL2RA locus, is not associated with either SLE or AAV. CONCLUSION: We have confirmed a previous suggestion that the IL2RA locus is associated with SLE and showed some evidence of association with AAV. Soluble IL-2RA concentrations correlate with rs11594656 genotype in quiescent disease in both AAV and SLE. Differential association of autoimmune diseases and SNPs within the IL2RA locus suggests that the IL2RA pathway may prove to play differing, as yet undefined, roles in each disease

Statin utilisation in a real‐world setting: a retrospective analysis in relation to arterial and cardiovascular autonomic function

Abstract Randomized trials suggest that statin treatment may lower blood pressure and influence cardiovascular autonomic function (CVAF), but the impact of duration of usage, discontinuation, and adherence to this therapy is unknown. We examined these issues with regard to blood pressure (BP)‐related variables in a large, population‐based study. Participants were 4942 adults (58% male; aged 50–84 years): 2179 on statin treatment and 2763 untreated. Days of utilization, adherence (proportion of days covered ≥0.8), and discontinuation (non‐use for ≥30 days immediately prior to BP measurement) of three statins (atorvastatin, pravastatin, and simvastatin) over a period of up to 2 years was monitored retrospectively from electronic databases. Systolic BP (SBP), diastolic BP (DBP), augmentation index, excess pressure, reservoir pressure, and CVAF (pulse rate and BP variability) parameters were calculated from aortic pressure waveforms derived from suprasystolic brachial measurement. Days of statin treatment had inverse relationships with pulse rate variability parameters in cardiac arrhythmic participants (20–25% lower than in statin non‐users) and with most arterial function parameters in everyone. For example, compared to untreated participants, those treated for ≥659 days had 3.0 mmHg lower aortic SBP (P < 0.01). Discontinuation was associated with higher brachial DBP and aortic DBP (for both, β = 2.0 mmHg, P = 0.008). Compared to non‐adherent statin users, adherent users had lower levels of brachial SBP, brachial DBP, aortic DBP, aortic SBP, and peak reservoir pressure (β = −1.4 to −2.6 mmHg). In conclusion, in a real‐world setting, statin‐therapy duration, non‐discontinuation and adherence associate inversely with BP variables and, in cardiac arrhythmias, CVAF parameters