Structural characterization of interaction of K11 and K63 mixed-linkage polyubiquitin chains with the tUIMs of epsin1

Ubiquitylation or covalent attachment of ubiquitin (Ub) to a variety of substrate proteins in cells is a versatile post-translational modification involved in the regulation of numerous cellular processes. The distinct messages that polyubiquitylation encodes are attributed to the multitude of conformations possible through attachment of ubiquitin monomers within a polyubiquitin chain via a specific lysine residue. Thus the hypothesis is that linkage defines polyubiquitin conformation which in turn determines specific recognition by cellular receptors. Ubiquitylation of membrane surface receptor proteins plays a very important role in regulating receptor-mediated endocytosis as well as endosomal sorting for lysosomal degradation. Epsin1 is an endocytic adaptor protein with three tandem UIMs (Ubiquitin Interacting Motifs) which are responsible for the highly specific interaction between epsin and ubiquitylated receptors. Epsin1 is also an oncogenic protein and its expression is upregulated in some types of cancer. Recently it has been shown that novel K11 and K63 mixed-linkage polyubiquitin chains serve as internalization signal for MHC I (Major Histocompatibility Complex I) molecule through their association with the tUIMs of epsin1. However the molecular mode of action and structural details of the interaction between polyubiquitin chains on receptors and tUIMs of epsin1 is yet to be determined. This information is crucial for the development of anticancer therapeutics targeting epsin1. The molecular basis for the linkage-specific recognition of K11 and K63 mixed-linkage polyubiquitin chains by the tandem UIMs of the endocytic adaptor protein epsin1 is investigated using a combination of NMR methods.

Previous land use and climate influence differences in soil organic carbon following reforestation of agricultural land with mixed-species plantings

Reforestation of agricultural land with mixed-species environmental plantings (native trees and shrubs) can contribute to mitigation of climate change through sequestration of carbon. Although soil carbon sequestration following reforestation has been investigated at site- and regional-scales, there are few studies across regions where the impact of a broad range of site conditions and management practices can be assessed. We collated new and existing data on soil organic carbon (SOC, 0–30 cm depth, N = 117 sites) and litter (N = 106 sites) under mixed-species plantings and an agricultural pair or baseline across southern and eastern Australia. Sites covered a range of previous land uses, initial SOC stocks, climatic conditions and management types. Differences in total SOC stocks following reforestation were significant at 52% of sites, with a mean rate of increase of 0.57 ± 0.06 Mg C ha−1 y−1. Increases were largely in the particulate fraction, which increased significantly at 46% of sites compared with increases at 27% of sites for the humus fraction. Although relative increase was highest in the particulate fraction, the humus fraction was the largest proportion of total SOC and so absolute differences in both fractions were similar. Accumulation rates of carbon in litter were 0.39 ± 0.02 Mg C ha−1 y−1, increasing the total (soil + litter) annual rate of carbon sequestration by 68%. Previously-cropped sites accumulated more SOC than previously-grazed sites. The explained variance differed widely among empirical models of differences in SOC stocks following reforestation according to SOC fraction and depth for previously-grazed (R2 = 0.18–0.51) and previously-cropped (R2 = 0.14–0.60) sites. For previously-grazed sites, differences in SOC following reforestation were negatively related to total SOC in the pasture. By comparison, for previously-cropped sites, differences in SOC were positively related to mean annual rainfall. This improved broad-scale understanding of the magnitude and predictors of changes in stocks of soil and litter C following reforestation is valuable for the development of policy on carbon markets and the establishment of future mixed-species environmental plantings.

Working with Student Teachers: A Mixed Methods Study to Examine the Roles and Self-Identified Dispositions of Cooperating Teachers

A critical component of teacher education is the field experience during which candidates practice under the supervision of experienced teachers. Programs use the InTASC Standards to define the requisite knowledge, skills, and dispositions for teaching. Practicing teachers are familiar with the concepts of knowledge and skills, but they are less familiar with dispositions. Practicing teachers who mentor prospective teachers are underrepresented in the literature, but they are critical to teacher preparation. The research goals were to describe the self-identified dispositions of cooperating teachers, identify what cooperating teachers consider their role in preparing prospective teachers, and explain challenges that cooperating teachers face. Using a mixed methods design, I conducted a quantitative survey followed by a qualitative case study. When I compared survey and case study data, cooperating teachers report possessing InTASC critical dispositions described in Standard 2: Learning Differences, Standard 3: Learning Environments, and Standard 9: Professional Learning and Ethical Practice, but not Standard 6: Assessment and Standard 10: Leadership and Collaboration. Cooperating teachers assume the roles of modeler, mentor and advisor, and informal evaluator. They explain student teachers often lack skills and dispositions to assume full teaching responsibilities and recommend that universities better prepare candidates for classrooms. Cooperating teachers felt university evaluations were not relevant to teaching reality. I recommend modifying field experiences to increase the quantity and duration of classroom placements. I suggest further research to detail cooperating teacher dispositions, compare cooperating teachers who work with different universities, and determine if cooperating teacher dispositions influence student teacher dispositions.

Using an end-of-life care pathway in acute stroke: a mixed methods study of decision-making and care experiences

Background: The evidence base on end-of-life care in acute stroke is limited, particularly with regard to recognising dying and related decision-making. There is also limited evidence to support the use of end-of-life care pathways (standardised care plans) for patients who are dying after stroke. Aim: This study aimed to explore the clinical decision-making involved in placing patients on an end-of-life care pathway, evaluate predictors of care pathway use, and investigate the role of families in decision-making. The study also aimed to examine experiences of end-of-life care pathway use for stroke patients, their relatives and the multi-disciplinary health care team. Methods: A mixed methods design was adopted. Data were collected in four Scottish acute stroke units. Case-notes were identified prospectively from 100 consecutive stroke deaths and reviewed. Multivariate analysis was performed on case-note data. Semi-structured interviews were conducted with 17 relatives of stroke decedents and 23 healthcare professionals, using a modified grounded theory approach to collect and analyse data. The VOICES survey tool was also administered to the bereaved relatives and data were analysed using descriptive statistics and thematic analysis of free-text responses. Results: Relatives often played an important role in influencing aspects of end-of-life care, including decisions to use an end-of-life care pathway. Some relatives experienced enduring distress with their perceived responsibility for care decisions. Relatives felt unprepared for and were distressed by prolonged dying processes, which were often associated with severe dysphagia. Pro-active information-giving by staff was reported as supportive by relatives. Healthcare professionals generally avoided discussing place of care with families. Decisions to use an end-of-life care pathway were not predicted by patients’ demographic characteristics; decisions were generally made in consultation with families and the extended health care team, and were made within regular working hours. Conclusion: Distressing stroke-related issues were more prominent in participants’ accounts than concerns with the end-of-life care pathway used. Relatives sometimes perceived themselves as responsible for important clinical decisions. Witnessing prolonged dying processes was difficult for healthcare professionals and families, particularly in relation to the management of persistent major swallowing difficulties.

A framework for mixed-type multi-outcome prediction with applications in healthcare

Health analysis often involves prediction of multiple outcomes of mixed-type. Existing work is restrictive to either a limited number or specific outcome types. We propose a framework for mixed-type multi-outcome prediction. Our proposed framework proposes a cumulative loss function composed of a specific loss function for each outcome type - as an example, least square (continuous outcome), hinge (binary outcome), poisson (count outcome) and exponential (non-negative outcome). Tomodel these outcomes jointly, we impose a commonality across the prediction parameters through a common matrix-Normal prior. The framework is formulated as iterative optimization problems and solved using an efficient Block coordinate descent method (BCD). We empirically demonstrate both scalability and convergence. We apply the proposed model to a synthetic dataset and then on two real-world cohorts: a Cancer cohort and an Acute Myocardial Infarction cohort collected over a two year period. We predict multiple emergency related outcomes - as example, future emergency presentations (binary), emergency admissions (count), emergency length-of-stay-days (non-negative) and emergency time-to-next-admission-day (non-negative). Weshow that the predictive performance of the proposed model is better than several state-of-the-art baselines.

Previous land use and climate influence differences in soil organic carbon following reforestation of agricultural land with mixed-species plantings

Reforestation of agricultural land with mixed-species environmental plantings (native trees and shrubs) can contribute to mitigation of climate change through sequestration of carbon. Although soil carbon sequestration following reforestation has been investigated at site- and regional-scales, there are few studies across regions where the impact of a broad range of site conditions and management practices can be assessed. We collated new and existing data on soil organic carbon (SOC, 0-30 cm depth, N = 117 sites) and litter (N = 106 sites) under mixed-species plantings and an agricultural pair or baseline across southern and eastern Australia. Sites covered a range of previous land uses, initial SOC stocks, climatic conditions and management types. Differences in total SOC stocks following reforestation were significant at 52% of sites, with a mean rate of increase of 0.57 ± 0.06 Mg C ha-1 y-1. Increases were largely in the particulate fraction, which increased significantly at 46% of sites compared with increases at 27% of sites for the humus fraction. Although relative increase was highest in the particulate fraction, the humus fraction was the largest proportion of total SOC and so absolute differences in both fractions were similar. Accumulation rates of carbon in litter were 0.39 ± 0.02 Mg C ha-1 y-1, increasing the total (soil + litter) annual rate of carbon sequestration by 68%. Previously-cropped sites accumulated more SOC than previously-grazed sites. The explained variance differed widely among empirical models of differences in SOC stocks following reforestation according to SOC fraction and depth for previously-grazed (R2 = 0.18-0.51) and previously-cropped (R2 = 0.14-0.60) sites. For previously-grazed sites, differences in SOC following reforestation were negatively related to total SOC in the pasture. By comparison, for previously-cropped sites, differences in SOC were positively related to mean annual rainfall. This improved broad-scale understanding of the magnitude and predictors of changes in stocks of soil and litter C following reforestation is valuable for the development of policy on carbon markets and the establishment of future mixed-species environmental plantings.

Concerns and worries in people living with inflammatory bowel disease (IBD): A mixed methods study.

OBJECTIVE: This mixed-methods study aimed to explore concerns and worries related to living with inflammatory bowel disease (IBD). METHODS: Overall, 294 patients with a clinically established diagnosis of IBD were enrolled in this cross-sectional study. Concerns and worries were measured with one open-ended question. Measures of anxiety and depressive symptoms and disease activity were also administered. A thematic analysis was conducted and thematic map created. Spearman's rho was used to identify univariate correlations between predictors and the main themes. Binary logistic regression was used to test the predictors of the main themes. RESULTS: Despite the majority of study participants being in IBD remission (74%, n=217), all but 11 reported significant IBD-related concerns. Twenty two percent reported symptoms of depression and 41% of anxiety. Four themes were identified: Quality of life (51%); Unpredictability (35%); Symptoms (34%) and Treatments (19%). Males and older people were less concerned about Quality of life (OR=.597, 95% CI: .363-.980 and OR=.980, 95% CI: .965-.995, respectively). Those in remission were less concerned about Symptoms (OR=.510, 95% CI: .281- .926) while those with longer disease duration worried more about the Symptoms (OR=1.035, 95% CI: 1.010-1.061). Males were less concerned about Treatments (OR=.422, 95% CI: .229-.777). CONCLUSION: IBD patients report significant disease-related concerns even when in remission. Further exploration of what predicts patient concerns may help in shaping health-care delivery so that it better addresses patient needs.

Organic management of Brachiaria brizantha cv. Marandu pasture mixed with Stylosanthes spp. in the Cerrado.

2016

Investigation of RuO2/Ta2O5 thin film evolution by thermogravimetry combined with mass spectrometry

The thermal evolution process of RuO2–Ta2O5/Ti coatings with varying noble metal content has been investigated under in situ conditions by thermogravimetry combined with mass spectrometry. The gel-like films prepared from alcoholic solutions of the precursor salts (RuCl3·3H2O, TaCl5) onto titanium metal support were heated in an atmosphere containing 20% O2 and 80% Ar up to 600 °C. The evolution of the mixed oxide coatings was followed by the mass spectrometric ion intensity curves. The cracking of retained solvent and the combustion of organic surface species formed were also followed by the mass spectrometric curves. The formation of carbonyl- and carboxylate-type surface species connected to the noble metal was identified by Fourier transform infrared emission spectroscopy. These secondary processes–catalyzed by the noble metal–may play an important role in the development of surface morphology and electrochemical properties. The evolution of the two oxide phases does not take place independently, and the effect of the noble metal as a combustion catalyst was proved.

Raman spectroscopic characteristics of phthalocyanine and naphthalocyanine in sandwich-type phthalocyaninato and porphyrinato rare earth complexes Part 5. – Raman spectroscopic characteristics of naphthalocyanine in mixed [tetrakis(4-tert-butylphenyl)porphyrinato]-(naphthalocyaninato) rare earth double-deckers

Raman spectra were recorded in the range 400–1800 cm−1 for a series of 15 mixed \[tetrakis(4-tert-butylphenyl)porphyrinato](2,3-naphthalocyaninato) rare earth double-deckers M(TBPP)(Nc) (M = Y; La–Lu except Pm) using laser excitation at 632.8 and 785 nm. Comparisons with bis(naphthalocyaninato) rare earth counterparts reveal that the vibrations of the metallonaphthalocyanine M(Nc) fragment dominate the Raman features of M(TBPP)(Nc). When excited with radiation of 632.8 nm, the most intense vibration appears at about 1595 cm−1, due to the naphthalene stretching. These complexes exhibit the marker Raman band for Nc•− as a medium-intense band in the range 1496–1507 cm−1, attributed to the coupling of pyrrole and aza stretching, while the marker Raman band of Nc2− in intermediate-valence Ce(TBPP)(Nc) appears as a strong band at 1493 cm−1 and is due to the isoindole stretchings. By contrast, when excited with radiation of 785 nm that is in close resonance with the main Q absorption band of the naphthalocyanine ligand, the ring radial vibrations at ca 680 and 735 cm−1 for MIII(TBPP)(Nc) are selectively intensified and are the most intense bands. For the cerium double-decker, the most intense vibration also acting as the marker Raman band of Nc2− appears at 1497 cm−1 with contributions from both pyrrole CC and aza CN stretches. The same vibrational modes show weak to medium intensity scattering at 1506–1509 cm−1 for MIII(TBPP)(Nc) and this is the marker Raman band of Nc•− when thus excited. The scatterings due to the Nc breathings, ring radial vibration, aza group stretchings, naphthalene stretchings, benzoisoindole stretchings and the coupling of pyrrole CC and aza CN stretchings in MIII(TBPP)(Nc) are all slightly blue shifted along with the decrease in rare earth ionic radius, confirming the effects of increased ring–ring interactions on the Raman characteristics of naphthalocyanine in the mixed ring double-deckers.

Water uptake of aerosols with a focus on seeded aerosols and instrumentation techniques

This thesis focuses on the volatile and hygroscopic properties of mixed aerosol species. In particular, the influence organic species of varying solubility have upon seed aerosols. Aerosol studies were conducted at the Paul Scherrer Institut Laboratory for Atmospheric Chemistry (PSI-LAC, Villigen, Switzerland) and at the Queensland University of Technology International Laboratory for Air Quality and Health (QUT-ILAQH, Brisbane, Australia). The primary measurement tool employed in this program was the Volatilisation and Hygroscopicity Tandem Differential Mobility Analyser (VHTDMA - Johnson et al. 2004). This system was initially developed at QUT within the ILAQH and was completely re-developed as part of this project (see Section 1.4 for a description of this process). The new VHTDMA was deployed to the PSI-LAC where an analysis of the volatile and hygroscopic properties of ammonium sulphate seeds coated with organic species formed from the photo-oxidation of á-pinene was conducted. This investigation was driven by a desire to understand the influence of atmospherically prevalent organics upon water uptake by material with cloud forming capabilities. Of particular note from this campaign were observed influences of partially soluble organic coatings upon inorganic ammonium sulphate seeds above and below their deliquescence relative humidity (DRH). Above the DRH of the seed increasing the volume fraction of the organic component was shown to reduce the water uptake of the mixed particle. Below the DRH the organic was shown to activate the water uptake of the seed. This was the first time this effect had been observed for á-pinene derived SOA. In contrast with the simulated aerosols generated at the PSI-LAC a case study of the volatile and hygroscopic properties of diesel emissions was undertaken. During this stage of the project ternary nucleation was shown, for the first time, to be one of the processes involved in formation of diesel particulate matter. Furthermore, these particles were shown to be coated with a volatile hydrophobic material which prevented the water uptake of the highly hygroscopic material below. This result was a first and indicated that previous studies into the hygroscopicity of diesel emission had erroneously reported the particles to be hydrophobic. Both of these results contradict the previously upheld Zdanovksii-Stokes-Robinson (ZSR) additive rule for water uptake by mixed species. This is an important contribution as it adds to the weight of evidence that limits the validity of this rule.

At-risk students and classroom engagement : listening to “voices of possibility and hope"

In my work with secondary school students who have disengaged from mainstream classrooms, I have often been surprised at the ways they enthusiastically engage with the projects on offer. They have demonstrated that, in apparent contradiction of their classroom behaviour, they still maintain hope in achieving a positive outcome from education. In a long-running schools-university project employing a “students-as-researchers” approach to investigating educational disadvantage, “at-risk” students have produced high quality results. Naturally, I wanted to know what it was about this sort of pedagogy that seemed to work for them. In this chapter, then, I outline the project and discuss some reasons for disengagement. I then address the features of the project that the participants themselves have identified as being instrumental in their re-engagement with formal education. Finally, I consider how these features may be transposed to maintaining the educational engagement of at-risk students in mainstream classrooms.

Raman spectroscopic study of the mixed anion mineral Yecoraite Bi5Fe3O9(Te4+O3)(Te6+O4)2•9H2O

Tellurates are rare minerals as the tellurate anion is readily reduced to the tellurite ion. Often minerals with both tellurate and tellurite anions in the mineral are found. An example of such a mineral containing tellurate and tellurite is yecoraite. Raman spectroscopy has been used to study this mineral, the exact structure of which is unknown. Two Raman bands at 796 and 808 cm-1 are assigned to the ν1 (TeO4)2- symmetric and ν3 (TeO3)2- antisymmetric stretching modes and Raman bands at 699 cm-1 are attributed to the the ν3 (TeO4)2- antisymmetric stretching mode and the band at 690 cm-1 to the ν1 (TeO3)2- symmetric stretching mode. The intense band at 465 cm-1 with a shoulder at 470 cm-1 is assigned the (TeO4)2- and (TeO3)2- bending modes. Prominent Raman bands are observed at 2878, 2936, 3180 and 3400 cm-1. The band at 3936 cm-1 appears quite distinct and the observation of multiple bands indicates the water molecules in the yecoraite structure are not equivalent. The values for the OH stretching vibrations listed provide hydrogen bond distances of 2.625 Å (2878 cm-1), 2.636 Å (2936 cm-1), 2.697 Å (3180 cm-1) and 2.798 Å (3400 cm-1). This range of hydrogen bonding contributes to the stability of the mineral. A comparison of the Raman spectra of yecoraite with that of tellurate containing minerals kuranakhite, tlapallite and xocomecatlite is made.

Kava Anxiety Depression Spectrum Study (KADSS): a mixed methods RCT using an aqueous extract of Piper methysticum

Objectives: To report on the design, significance and potential impacts of the first documented human clinical trial assessing the anxiolytic and thymoleptic efficacy of an aqueous monoextract of Piper methysticum (kava). The significance of the qualitative element of our clinical trial is also explored. The Kava Anxiety Depression Spectrum Study (KADSS) is a 3-week placebocontrolled, double-blind, cross-over trial involving 60 adult participants (18—65) with elevated stable anxiety and varying levels of depressive symptoms. Aims: The aims of KADSS are: (1) to determine whether an aqueous standardised extract of kava is effective for the treatment of anxiety; (2) to assess the effects of kava on differing levels of depression; and (3) to explore participants’ experience of taking kava via qualitative research. The study also provides preliminary assessment of the safety of an aqueous extract of kava in humans. Conclusion: If results reveal that the aqueous kava preparation exerts significant anxiolytic effects and appears safe, potentially beneficial impacts may occur. Data supporting a safe and effective kava extract may encourage a re-introduction of kava to Europe, UK and Canada. This may provide a major socioeconomic benefit to Pacific Island nations, and to sufferers of anxiety disorders.

Raman spectroscopic study of the mixed anion sulphate-arsenate mineral parnauite Cu9[(OH)10|SO4|(AsO4)2].7H2O

The mixed anion mineral parnauite Cu9[(OH)10|SO4|(AsO4)2].7H2O has been studied by Raman spectroscopy. Characteristic bands associated with arsenate, sulphate, hydroxyl units are identified. Broad bands are observed and are resolved into component bands. Two intense bands at 859 and 830 cm-1 are assigned to the 1 (AsO4)3- symmetric stretching and 3 (AsO4)3- antisymmetric stretching modes. The comparatively sharp band at 976 cm-1 is assigned to the ν1 (SO4)2- symmetric stretching mode and a broad spectral profile centered upon 1097 cm-1 is attributed to the ν3 (SO4)2- antisymmetric stretching mode. A comparison of the Raman spectra is made with other arsenate bearing minerals such as carminite, clinotyrolite, kankite, tilasite and pharmacosiderite.