Genomic organisation, activity and distribution analysis of the microbial putrescine oxidase degradation pathway

The catalytic action of putrescine specific amine oxidases acting in tandem with 4-aminobutyraldehyde dehydrogenase is explored as a degradative pathway in Rhodococcus opacus. By limiting the nitrogen source, increased catalytic activity was induced leading to a coordinated response in the oxidative deamination of putrescine to 4-aminobutyraldehyde and subsequent dehydrogenation to 4-aminobutyrate. Isolating the dehydrogenase by ion exchange chromatography and gel filtration revealed that the enzyme acts principally on linear aliphatic aldehydes possessing an amino moiety. Michaelis-Menten kinetic analysis delivered a Michaelis constant (KM=0.014mM) and maximum rate (Vmax=11.2μmol/min/mg) for the conversion of 4-aminobutyraldehyde to 4-aminobutyrate. The dehydrogenase identified by MALDI-TOF mass spectrometric analysis (E value=0.031, 23% coverage) belongs to a functionally related genomic cluster that includes the amine oxidase, suggesting their association in a directed cell response. Key regulatory, stress and transport encoding genes have been identified, along with candidate dehydrogenases and transaminases for the further conversion of 4-aminobutyrate to succinate. Genomic analysis has revealed highly similar metabolic gene clustering among members of Actinobacteria, providing insight into putrescine degradation notably among Micrococcaceae, Rhodococci and Corynebacterium by a pathway that was previously uncharacterised in bacteria.

Identification, functional expression and kinetic analysis of two primary amine oxidases from Rhodococcus opacus

Two native copper-containing amine oxidases (EC 1.4.3.21) have been isolated from Rhodococcus opacus and reveal phenotypic plasticity and catalytic activity with respect to structurally diverse natural and synthetic amines. Altering the amine growth substrate has enabled tailored and targeted oxidase upreg-ulation, which with subsequent treatment by precipitation, ion exchange and gel filtration, achieved a 90–150 fold purification. MALDI-TOF mass spectrometric and genomic analysis has indicated multiple gene activation with complex biodegradation pathways and regulatory mechanisms. Additional post-purification characterisation has drawn on the use of carbonyl reagent and chelating agent inhibitors. Michaelis–Menten kinetics for common aliphatic and aromatic amine substrates and several structural analogues demonstrated a broad specificity and high affinity with Michaelis constants (K M) ranging from 0.1 to 0.9 mM for C 1 –C 5 aliphatic mono-amines and <0.2 mM for a range of aromatic amines. Potential exploitation of the enzymatic versatility of the two isolated oxidases in biosensing and bioprocessing is discussed.

Role of amine oxidase expression to maintain putrescine homeostasis in Rhodococcus opacus

While applications of amine oxidases are increasing, few have been characterised and our understanding of their biological role and strategies for bacteria exploitation are limited. By altering the nitrogen source (NH4Cl, putrescine and cadaverine (diamines) and butylamine (monoamine)) and concentration, we have identified a constitutive flavin dependent oxidase (EC 1.4.3.10) within Rhodococcus opacus. The activity of this oxidase can be increased by over two orders of magnitude in the presence of aliphatic diamines. In addition, the expression of a copper dependent diamine oxidase (EC 1.4.3.22) was observed at diamine concentrations>1mM or when cells were grown with butylamine, which acts to inhibit the flavin oxidase. A Michaelis-Menten kinetic treatment of the flavin oxidase delivered a Michaelis constant (KM)=190μM and maximum rate (kcat)=21.8s(-1) for the oxidative deamination of putrescine with a lower KM (=60μM) and comparable kcat (=18.2s(-1)) for the copper oxidase. MALDI-TOF and genomic analyses have indicated a metabolic clustering of functionally related genes. From a consideration of amine oxidase specificity and sequence homology, we propose a putrescine degradation pathway within Rhodococcus that utilises oxidases in tandem with subsequent dehydrogenase and transaminase enzymes. The implications of PUT homeostasis through the action of the two oxidases are discussed with respect to stressors, evolution and application in microbe-assisted phytoremediation or bio-augmentation.

A molecular phylogeny of the checkered beetles and a description of Epiclininae a new subfamily (Coleoptera: Cleroidea: Cleridae)

We provide the first molecular phylogeny of the clerid lineage (Coleoptera: Cleridae, Thanerocleridae) within the superfamily Cleroidea to examine the two most recently-proposed hypotheses of higher-level classification. Phylogenetic relationships of checkered beetles were inferred from approximately ~5,000nt of both nuclear and mitochondrial rDNA (28S, 16S, and 12S) and the mitochondrial protein-coding gene COI. A worldwide sample of ~70 genera representing almost a quarter of generic diversity of the clerid lineage was included and phylogenies were reconstructed using Bayesian and Maximum Likelihood approaches. Results support the monophyly of many proposed subfamilies but were not entirely congruent with either current classification system. The subfamilial relationships within the Cleridae are resolved with support for three main lineages. Tillinae are supported as the sister group to all other subfamilies within the Cleridae, whereas Thaneroclerinae, Korynetinae and a new subfamily formally described here, Epiclininae subf. n, form a sister group to Clerinae + Hydnocerinae.

Understanding the information literacy experiences of EFL (English as a Foreign Language) students

This thesis investigated the information literacy experiences of EFL (English as a Foreign Language) students in a higher education institution in the United Arab Emirates (UAE). Phenomenography was used to investigate how EFL students' 'used information to learn' (ie. information literacy). The study revealed that EFL students' experienced information literacy across four categories and had varying experiences of information and learning. The research also showed that EFL students' faced a number of challenges and barriers due to language that impacted on their experiences of reading, understanding, accessing and translating information.

Herbivores and nutrients control grassland plant diversity via light limitation

Human alterations to nutrient cycles1, 2 and herbivore communities3, 4, 5, 6, 7 are affecting global biodiversity dramatically2. Ecological theory predicts these changes should be strongly counteractive: nutrient addition drives plant species loss through intensified competition for light, whereas herbivores prevent competitive exclusion by increasing ground-level light, particularly in productive systems8, 9. Here we use experimental data spanning a globally relevant range of conditions to test the hypothesis that herbaceous plant species losses caused by eutrophication may be offset by increased light availability due to herbivory. This experiment, replicated in 40 grasslands on 6 continents, demonstrates that nutrients and herbivores can serve as counteracting forces to control local plant diversity through light limitation, independent of site productivity, soil nitrogen, herbivore type and climate. Nutrient addition consistently reduced local diversity through light limitation, and herbivory rescued diversity at sites where it alleviated light limitation. Thus, species loss from anthropogenic eutrophication can be ameliorated in grasslands where herbivory increases ground-level light.

Rad51 supports triple negative breast cancer metastasis

In contrast to extensive studies on familial breast cancer, it is currently unclear whether defects in DNA double strand break (DSB) repair genes play a role in sporadic breast cancer development and progression. We performed analysis of immunohistochemistry in an independent cohort of 235 were sporadic breast tumours. This analysis suggested that RAD51 expression is increased during breast cancer progression and metastasis and an oncogenic role for RAD51 when deregulated. Subsequent knockdown of RAD51 repressed cancer cell migration in vitro and reduced primary tumor growth in a syngeneic mouse model in vivo. Loss of RAD51 also inhibited associated metastasis not only in syngeneic mice but human xenografts and changed the metastatic gene expression profile of cancer cells, consistent with inhibition of distant metastasis. This demonstrates for the first time a new function of RAD51 that may underlie the proclivity of patients with RAD51 overexpression to develop distant metastasis. RAD51 is a potential biomarker and attractive drug target for metastatic triple negative breast cancer, with the capability to extend the survival of patients, which is less than 6 months.

Foreword

Foreword for the book, "INTERCHANGING: Future designs for responsive transport environments"

Urban Informatics and Public Transport

The holistic urban experience we perceive when immersed in an urban context is at the heart of urban informatics. This experience encompasses all urban elements such as architecture, people, and culture. Urban informatics explores the possibilities and opportunities created by new technologies and information for enhancing the urban experience. Public transport is an essential urban experience. Everyday, urban dwellers takes public transport to commute and move between different parts of the city. Public transport serves people from all over the city and moves them through different places in the city, using different means of transportation. The nature of public transport—involving people, places, and technologies, makes it a fitting context for urban informatics interventions. There are three main aspects of the public transport experience that can readily benefit from urban informatics interventions the: pragmatic aspect, hedonistic aspect, and social aspect. From the pragmatic perspective, these interventions can help people to be more efficient and effective in taking public transport. Hedonistic-related interventions aim to bring enjoyment and fun to our mundane commute. Finally, urban informatics can strengthen the sense of community in a socially-passive context like public transport environments through adopting socially focused interventions.

Regulation of microRNA-1288 in colorectal cancer : altered expression and its clinicopathological significance

We aim to examine the miR-1288 expression in cancer cell lines and a large cohort of patients with colorectal cancer. Two colon cancer cell lines (SW480 and SW48) and one normal colonic epithelial cell line (FHC) were recruited. The miRNA expressions of miR-1288 were tested on these cell lines by using quantitative real-time polymerase chain reaction (qRT-PCR). An exogenous miR-1288 (mimic) was used to detect cell proliferation and cell cycle changes in SW480 using MTT calorimetric assay and flow cytometry, respectively. In addition, tissues from 122 patients with surgical resection of colorectum (82 adenocarcinomas, 20 adenomas, and 20 non-neoplastic tissues) were tested for miR-1288 expression by qRT-PCR. The colon cancer cell lines showed reduced expression of miR-1288 compared to normal colonic epithelial cell line. Over expression of miR-1288 in SW480 cell line showed increased cell proliferation and increased G2-M phase cells. In tissues, reduced miR-1288 expression was noted in majority of colorectal adenocarcinoma compared to colorectal adenoma and non-neoplastic tissues. Reduced or absent expression of miR-1288 was noted in 76% (n = 62/82) of the cancers. The expression levels of miR-1288 were higher in distal colorectal adenocarcinomas (P = 0.013) and in cancers of lower T staging (P = 0.033). To conclude, alternation of miR-1288 expression is important in the progression of colorectal cancer. The differential regulation of miR-1288 was found to be related to cancer location and pathological staging in colorectal cancers.

Gentrification and the artistic dividend : the role of the arts in neighborhood change

Problem, research strategy, and findings: There is a conflict between recent creative placemaking policies intended to promote positive neighborhood development through the arts and the fact that the arts have long been cited as contributing to gentrification and the displacement of lower-income residents. Unfortunately, we do not have data to demonstrate widespread evidence of either outcome. We address the dearth of comprehensive research and inform neighborhood planning efforts by statistically testing how two different groups of arts activities—the fine arts and commercial arts industries—are associated with conditions indicative of revitalization and gentrification in 100 large U.S. metropolitan areas. We find that different arts activities are associated with different types and levels of neighborhood change. Commercial arts industries show the strongest association with gentrification in rapidly changing areas, while the fine arts are associated with stable, slow-growth neighborhoods. Takeaway for practice: This research can help planners to more effectively incorporate the arts into neighborhood planning efforts and to anticipate the potential for different outcomes in their arts development strategies, including gentrification-related displacement.

The location patterns of artistic clusters : a metro- and neighborhood-level analysis

Analysing census and industry data at the metro and neighbourhood levels, this paper seeks to identify the location characteristics associated with artistic clusters and determine how these characteristics vary across different places. We find that the arts cannot be taken overall as an urban panacea, but rather that their impact is place-specific and policy ought to reflect these nuances. However, our work also finds that, paradoxically, the arts’ role in developing metro economies is as highly underestimated as it is overgeneralised. While arts clusters exhibit unique industry, scale and place-specific attributes, we also find evidence that they cluster in ‘innovation districts’, suggesting they can play a larger role in economic development. To this end, our results raise important questions and point toward new approaches for arts-based urban development policy that look beyond a focus on the arts as amenities to consider the localised dynamics between the arts and other industries.

Art Spaces, Art Places : Examining Neighborhood Preferences of New York Arts Organizations

This report builds on and extends a diverse literature that examines the location patterns of the arts and creative industries through analysis of a database of arts nonprofit organizations from the New York State Cultural Data Project. We confirm the link between arts organizations and the urban core and creative economy, but challenge the assumption that arts tend to locate in ethnic and disadvantaged neighborhoods. By identifying key neighborhood attributes associated with distinct types of arts organizations, we can better identify potential sites conducive to nurturing additional artistic activity and inform strategies to engage organizations in neighborhoods that are underserved in the arts.

Understanding the information literacy experiences of EFL (English as a Foreign Language) students

Purpose – This paper outlines research that explores the information literacy experiences of EFL (English as a Foreign Language) students. The question explored in this research was: how do EFL students experience information literacy. Design/Methodology/Approach – This study used phenomenography, a relational approach to explore the information literacy experiences of EFL students. Phenomenography studies the qualitatively different ways a phenomenon is experienced in the world around us. Findings – This research revealed that EFL students experienced information literacy in four qualitatively different ways. The four categories revealed through the data were: process, quality, language and knowledge. This research found that language impacted on EFL students’ experiences of information literacy and revealed that EFL students applied various techniques and strategies when they read, understood, organised and translated information. Research limitations/implications – This research was conducted in a specific cultural and educational context, therefore the results might not reflect the experiences of EFL students in other cultural or educational contexts. Practical implications – The findings from this research offer an important contribution to information literacy practice by providing important insights about EFL students’ experiences and perceptions of information and learning that can be used to inform curriculum development in second language learning contexts. Originality/Value - There is currently a lack of research using a relational approach to investigate EFL students’ experiences of information literacy. There is also limited research that explores the impact language has on information literary and learning in English as a foreign or second language contexts.

Towards a phylogeny of the Tenebrionoidea (Coleoptera)

The phylogenetic relationships of the beetle superfamily Tenebrionoidea are investigated using the most comprehensive genetic data set compiled to date. With ∼34,000 described species in approximately 1250 genera and 28 families, Tenebrionoidea represent one of the most diverse and species-rich superfamilies of beetles. The interfamilial relationships of the Tenebrionoidea are poorly known; previous morphological and molecular phylogenies recovered few well-supported and often conflicting relationships between families. Here we present a molecular phylogeny of Tenebrionoidea based on genes commonly used to resolve family and superfamily-level phylogenies of beetles (18S, 28S, 16S, 12S, tRNA Val and COI). The alignment spanned over 6.5 KB of DNA sequence and over 300 tenebrionoid genera from 24 of the 28 families were sampled. Maximum Likelihood and Bayesian analysis could not resolve deeper level divergences within the superfamily and very few relationships between families were supported. Increasing gene coverage in the alignment by removing taxa with missing data did not improve clade support but when rogue taxa were removed increased resolution was recovered. Investigation of signal strength suggested conflicting phylogenetic signal was present in the standard genes used for beetle phylogenetics, even when rogue taxa were removed. Our study of Tenebrionoidea highlights that even with relatively comprehensive taxon sampling within a lineage, this standard set of genes is unable to resolve relationships within this superfamily.