Occurrence of LINE, gypsy-like, and copia-like retrotransposons in the clonally propagated sweet potato (Ipomoea batatas L.)

Retrotransposons are a class of transposable elements that represent a major fraction of the repetitive DNA of most eukaryotes. Their abundance stems from their expansive replication strategies. We screened and isolated sequence fragments of long terminal repeat (LTR), gypsy-like reverse transcriptase (rt) and gypsy-like envelope (env) domains, and two partial sequences of non-LTR retrotransposons, long interspersed element (LINE), in the clonally propagated allohexaploid sweet potato (Ipomoea batatas (L.) Lam.) genome. Using dot-blot hybridization, these elements were found to be present in the ~1597 Mb haploid sweet potato genome with copy numbers ranging from ~50 to ~4100 as observed in the partial LTR (IbLtr-1) and LINE (IbLi-1) sequences, respectively. The continuous clonal propagation of sweet potato may have contributed to such a multitude of copies of some of these genomic elements. Interestingly, the isolated gypsy-like env and gypsy-like rt sequence fragments, IbGy-1 (~2100 copies) and IbGy-2 (~540 copies), respectively, were found to be homologous to the Bagy-2 cDNA sequences of barley (Hordeum vulgare L.). Although the isolated partial sequences were found to be homologous to other transcriptionally active elements, future studies are required to determine whether they represent elements that are transcriptionally active under normal and (or) stressful conditions.

Re-conceptualising and re-positioning Australian library and information science education for the 21st century [Final Report 2011]

How can Australian library and information science (LIS) education produce, in a sustainable manner, the diverse supply of graduates with the appropriate attributes to develop and maintain high quality professional practice in the rapidly changing 21st century? This report presents the key findings of a project that has examined this question through research into future directions for LIS education in Australia. Titled Re-conceptualising and re-positioning Australian library and information science education for the twenty-first century, the purpose of the project was to establish a consolidated and holistic picture of the Australian LIS profession, and identify how its future education and training can be mediated in a cohesive and sustainable manner. The project was undertaken with a team of 12 university and vocational LIS educators from 11 institutions around Australia between November 2009 and December 2010. Collectively, these eleven institutions represented the broad spectrum and diversity of LIS education in Australia, and enabled the project to examine education for the information profession in a holistic and synergistic manner. Participating institutions in the project included Queensland University of Technology (Project Leader), Charles Sturt University, Curtin University of Technology, Edith Cowan University, Monash University, RMIT University, University of Canberra, University of South Australia, University of Tasmania, University of Technology Sydney and Victoria University. The inception and need for the project was motivated by a range of factors. From a broad perspective several of these factors relate to concerns raised at national and international levels regarding problems with education for LIS. In addition, the motivation and need for the project also related to some unique challenges that LIS education faces in the Australian tertiary education landscape. Over recent years a range of responses to explore the various issues confronting LIS education in Australia have emerged at local and national levels however this project represented the first significant investment of funding for national research in this area. In this way, the inception of the project offered a unique opportunity and powerful mechanism through which to bring together key stakeholders and inspire discourse concerning future education for the profession. Therefore as the first national project of its kind, its intent has been to provide foundation research that will inform and guide future directions for LIS education and training in Australia. The primary objective of the project was to develop a Framework for the Education of the Information Professions in Australia. The purpose of this framework was to provide evidence based strategic recommendations that would guide Australia’s future education for the information professions. Recognising the three major and equal players in the education process the project was framed around three areas of consideration: LIS students, the LIS workforce and LIS educators. Each area of consideration aligned to a research substudy in the project. The three research substudies were titled Student Considerations, Workforce Planning Considerations and Tertiary Education Considerations. The Students substudy provided a profile of LIS students and an analysis of their choices, experiences and expectations in regard to LIS education and their graduate destinations. The Workforce substudy provided an overview and analysis of the nature of the current LIS workforce, including a focus on employer expectations and employment opportunities and comment on the core and elective skill, knowledge and attitudes of current and future LIS professionals. Finally the Tertiary Education substudy provided a profile of LIS educators and an analysis of their characteristics and experiences including the key issues and challenges. In addition it also explored current national and international trends and priorities impacting on LIS education. The project utilised a Community Based Participatory Research (CBPR) approach. This approach involves all members of the community in all aspects of the project. It recognised the unique strengths and perspectives that community members bring to the process. For this project ‘community’ comprised of all individuals who have a role in, or a vested interest in, LIS education and included LIS educators, professionals, employers, students and professional associations. Individuals from these sub-groups were invited to participate in a range of aspects of the project from design through to implementation and evaluation. A range of research methodologies were used to consider the many different perspectives of LIS education, including employers and recruiters, professional associations, students, graduates and LIS teaching staff. Data collection involved a mixed method approach of questionnaires, focus groups, semi-structured interviews and environmental scans. An array of approaches was selected to ensure that broadest possible access to different facets of the information profession would be achieved. The main findings and observations from each substudy have highlighted a range of challenges for LIS education that need to be addressed. These findings and observations have grounded the development of the Framework for the Education of the Information Professions in Australia. The framework presents eleven recommendations to progress the national approach to LIS education and guide Australia’s future education for the information professions. The framework will be used by the LIS profession, most notably its educators, as strategic directions for the future of LIS education in Australia. Framework for the Education of the Information Professions in Australia: Recommendation 1: It is recommended that a broader and more inclusive vocabulary be adopted that both recognises and celebrates the expanding landscape of the field, for example ‘information profession’, ‘information sector’, ‘information discipline’ and ‘information education’. Recommendation 2: It is recommended that a self-directed body composed of information educators be established to promote, support and lead excellence in teaching and research within the information discipline. Recommendation 3: It is recommended that Australia’s information discipline continue to develop excellence in information research that will raise the discipline’s profile and contribute to its prominence within the national and international arena. Recommendation 4: It is recommended that further research examining the nature and context of Australia’s information education programs be undertaken to ensure a sustainable and relevant future for the discipline. Recommendation 5: It is recommended that further research examining the pathways and qualifications available for entry into the Australian information sector be undertaken to ensure relevance, attractiveness, accessibility and transparency. Recommendation 6: It is recommended that strategies are developed and implemented to ensure the sustainability of the workforce of information educators. Recommendation 7: It is recommended that a national approach to promoting and marketing the information profession and thereby attracting more students to the field is developed. Recommendation 8: It is recommended that Australia’s information discipline continues to support a culture of quality teaching and learning, especially given the need to accommodate a focus on the broader information landscape and more flexible delivery options. Recommendation 9: It is recommended that strategies are developed that will support and encourage collaboration between information education within the higher education and VET sectors. Recommendation 10: It is recommended that strategies and forums are developed that will support the information sector working together to conceptualise and articulate their professional identity and educational needs. Recommendation 11: It is recommended that a research agenda be established that will identify and prioritise areas in which further development or work is needed to continue advancing information education in Australia. The key findings from this project confirm that a number of pressing issues are confronting LIS education in Australia. Left unaddressed these issues will have significant implications for the future of LIS education as well as the broader LIS profession. Consequently creating a sustainable and cohesive future can only be realised through cooperation and collaboration among all stakeholders including those with the capacity to enact radical change in university and vocational institutions. Indeed the impending adoption and implementation of the project’s recommendations will fundamentally determine whether Australian LIS education is assured both for the present day and into the future.

Expression Pattern of the Alpha-Kafirin Promoter Coupled with a Signal Peptide from Sorghum bicolor L. Moench

Regulatory sequences with endosperm specificity are essential for foreign gene expression in the desired tissue for both grain quality improvement and molecular pharming. In this study, promoters of seed storage α-kafirin genes coupled with signal sequence (ss) were isolated from Sorghum bicolor L. Moench genomic DNA by PCR. The α-kafirin promoter (α-kaf) contains endosperm specificity-determining motifs, prolamin-box, the O2-box 1, CATC, and TATA boxes required for α-kafirin gene expression in sorghum seeds. The constructs pMB-Ubi-gfp and pMB-kaf-gfp were microprojectile bombarded into various sorghum and sweet corn explants. GFP expression was detected on all explants using the Ubi promoter but only in seeds for the α-kaf promoter. This shows that the α-kaf promoter isolated was functional and demonstrated seed-specific GFP expression. The constructs pMB-Ubi-ss-gfp and pMB-kaf-ss-gfp were also bombarded into the same explants. Detection of GFP expression showed that the signal peptide (SP)::GFP fusion can assemble and fold properly, preserving the fluorescent properties of GFP.

Development of a transformation system for sorghum (Sorghum bicolor L.)

Sorghum (Sorghum bicolor (L.) Moench) is the world’s fifth major cereal crop and holds importance as a construction material, food and fodder source. More recently, the potential of this plant as a biofuel source has been noted. Despite its agronomic importance, the use of sorghum production is being constrained by both biotic and abiotic factors. These challenges could be addressed by the use of genetic engineering strategies to complement conventional breeding techniques. However, sorghum is one of the most recalcitrant crops for genetic modification with the lack of an efficient tissue culture system being amongst the chief reasons. Therefore, the aim of this study was to develop an efficient tissue culture system for establishing regenerable embryogenic cell lines, micropropagation and acclimatisation for Sorghum bicolor and use this to optimise parameters for genetic transformation via Agrobacterium-mediated transformation and microprojectile bombardment. Using five different sorghum cultivars, SA281, 296B, SC49, Wray and Rio, numerous parameters were investigated in an attempt to establish an efficient and reproducible tissue culture and transformation system. Using immature embryos (IEs) as explants, regenerable embryogenic cell lines (ECLs) could only be established from cultivars SA281 and 296B. Large amounts of phenolics were produced from IEs of cultivars, SC49, Wary and Rio, and these compounds severely hindered callus formation and development. Cultivar SA281 also produced phenolics during regeneration. Attempts to suppress the production of these compounds in cultivars SA281 and SC49 using activated charcoal, PVP, ascorbic acid, citric acid and liquid filter paper bridge methods were either ineffective or had a detrimental effect on embryogenic callus formation, development and regeneration. Immature embryos sourced during summer were found to be far more responsive in vitro than those sourced during winter. In an attempt to overcome this problem, IEs were sourced from sorghum grown under summer conditions in either a temperature controlled glasshouse or a growth chamber. However, the performance of these explants was still inferior to that of natural summer-sourced explants. Leaf whorls, mature embryos, shoot tips and leaf primordia were found to be unsuitable as explants for establishing ECLs in sorghum cultivars SA281 and 296B. Using the florets of immature inflorescences (IFs) as explants, however, ECLs were established and regenerated for these cultivars, as well as for cultivar Tx430, using callus induction media, SCIM, and regeneration media, VWRM. The best in vitro responses, from the largest possible sized IFs, were obtained using plants at the FL-2 stage (where the last fully opened leaf was two leaves away from the flag leaf). Immature inflorescences could be stored at 25oC for up to three days without affecting their in vitro responses. Compared to IEs, the IFs were more robust in tissue culture and showed responses which were season and growth condition independent. A micropropagation protocol for sorghum was developed in this study. The optimum plant growth regulator (PGR) combination for the micropropagation of in vitro regenerated plantlets was found to be 1.0 mg/L BAP in combination with 0.5 mg/L NAA. With this protocol, cultivars 296B and SA281 produced an average of 57 and 13 off-shoots per plantlet, respectively. The plantlets were successfully acclimatised and developed into phenotypically normal plants that set seeds. A simplified acclimatisation protocol for in vitro regenerated plantlets was also developed. This protocol involved deflasking in vitro plantlets with at least 2 fully-opened healthy leaves and at least 3 roots longer than 1.5 cm, washing the media from the roots with running tap water, planting in 100 mm pots and placing in plastic trays covered with a clear plastic bag in a plant growth chamber. After seven days, the corners of the plastic cover were opened and the bags were completely removed after 10 days. All plantlets were successfully acclimatised regardless of whether 1:1 perlite:potting mix, potting mix, UC mix or vermiculite were used as potting substrates. Parameters were optimised for Agrobacterium-mediated transformation (AMT) of cultivars SA281, 296B and Tx430. The optimal conditions were the use of Agrobacterium strain LBA4404 at an inoculum density of 0.5 OD600nm, heat shock at 43oC for 3 min, use of the surfactant Pluronic F-68 (0.02% w/v) in the inoculation media with a pH of 5.2 and a 3 day co-cultivation period in dark at 22oC. Using these parameters, high frequencies of transient GFP expression was observed in IEs precultured on callus initiation media for 1-7 days as well as in four weeks old IE- and IF-derived callus. Cultivar SA281 appeared very sensitive to Agrobacterium since all tissue turned necrotic within two weeks post-exposure. For cultivar 296B, GFP expression was observed up to 20 days post co-cultivation but no stably transformed plants were regenerated. Using cultivar Tx430, GFP was expressed for up to 50 days post co-cultivation. Although no stably transformed plants of this cultivar were regenerated, this was most likely due to the use of unsuitable regeneration media. Parameters were optimised for transformation by particle bombardment (PB) of cultivars SA281, 296B and Tx430. The optimal conditions were use of 3-7 days old IEs and 4 weeks old IF callus, 4 hour pre- and post-bombardment osmoticum treatment, use of 0.6 µm gold microparticles, helium pressure of 1500 kPa and target distance of 15 cm. Using these parameters for PB, transient GFP expression was observed for up to 14, 30 and 50 days for cultivars SA281, 296B and Tx430, respectively. Further, the use of PB resulted in less tissue necrosis compared to AMT for the respective cultivars. Despite the presence of transient GFP expression, no stably transformed plants were regenerated. The establishment of regenerable ECLs and the optimization of AMT and PB parameters in this study provides a platform for future efforts to develop an efficient transformation protocol for sorghum. The development of GM sorghum will be an important step towards improving its agronomic properties as well as its exploitation for biofuel production.

Aromatic l-amino acid decarboxylase : histochemical localization in rat kidney and lack of effect of dietary potassium or sodium loading on enzyme distribution

Utilizing a mono-specific antiserum produced in rabbits to hog kidney aromatic L-amino acid decarboxylase (AADC), the enzyme was localized in rat kidney by immunoperoxidase staining. AADC was located predominantly in the proximal convoluted tubules; there was also weak staining in the distal convoluted tubules and collecting ducts. An increase in dietary potassium or sodium intake produced no change in density or distribution of AADC staining in kidney. An assay of AADC enzyme activity showed no difference in cortex or medulla with chronic potassium loading. A change in distribution or activity of renal AADC does not explain the postulated dopaminergic modulation of renal function that occurs with potassium or sodium loading.

Piecing together an integrative taxonomic puzzle: microsatellite, wing shape and aedeagus length analyses of Bactrocera dorsalis s.l. (Diptera: Tephritidae) find no evidence of multiple lineages in a proposed contact zone along the Thai/Malay Peninsula

Bactrocera dorsalis (Hendel) and B. papayae Drew & Hancock represent a closely related sibling species pair for which the biological species limits are unclear; i.e., it is uncertain if they are truely two biological species, or one biological species which has been incorrectly taxonomically split. The geographic ranges of the two taxa are thought to abut or overlap on or around the Isthmus of Kra, a recognised biogeographic barrier located on the narrowest portion of the Thai Peninsula. We collected fresh material of B. dorsalis sensu lato (i.e., B. dorsalis sensu stricto + B. papayae) in a north-south transect down the Thai Peninsula, from areas regarded as being exclusively B. dorsalis s.s., across the Kra Isthmus, and into regions regarded as exclusively B. papayae. We carried out microsatellite analyses and took measurements of male genitalia and wing shape. Both the latter morphological tests have been used previously to separate these two taxa. No significant population structuring was found in the microsatellite analysis and results were consistent with an interpretation of one, predominantly panmictic population. Both morphological datasets showed consistent, clinal variation along the transect, with no evidence for disjunction. No evidence in any tests supported historical vicariance driven by the Isthmus of Kra, and none of the three datasets supported the current taxonomy of two species. Rather, within and across the area of range overlap or abutment between the two species, only continuous morphological and genetic variation was recorded. Recognition that morphological traits previously used to separate these taxa are continuous, and that there is no genetic evidence for population segregation in the region of suspected species overlap, is consistent with a growing body of literature that reports no evidence of biological differentiation between these taxa.

Effect of a novel compound from Lycopodium obscurum L. on osteogenic activity of osteoblasts in vitro

We investigated the potential of an extract of Lycopodium obscurum L.; stigmastane-3-oxo-21-oic acid (SA), to enhance osteogensis of mouse osteoblastic MC3T3-E1 cells. SA at a concentration of 16 µM was found to have no significant effect upon the viability of the cells, thus concentrations of 8 µM and 16 µM of SA were used in all further experiments. Both concentrations of SA had an inhibitory affect upon alkaline phosphatase activity (ALP) after 8 days incubation, however, after 16 days activity was restored to control levels. However Alizarin red S staining showed increased levels of mineralization for both concentrations after 16 days culture. Real time PCR showed inhibition of genes Runx2 and Osterix genes responsible for the up-regulation of ALP. However early time point (8 days) up-regulation of bone matrix mineralization genes OPN and OCN, and late time point (16 days) up-regulation of both Jun-D and Fra-2 mRNA expression was significantly enhanced. These results suggest a potential me-chanism of SA in enhancing bone fracture healing is through the up-regulating bone matrix minera-lization.

Teaching and learning family and consumer sciences through K-W-L charts

While much of the control and many of the activities found in today’s classrooms have been placed in the hands of the learners and learning has become inquiry-based, there remains a need for teachers to use teaching tools that would facilitate this student-centered teaching process. This article identifies the K-W-L Chart as one such tool and follows a case study of four Kuwaiti ‘Family and Consumer Sciences’ teaching / learning events to evaluate their ability to enhance the learning outcomes of eight students. The research was designed from a qualitative, multi-tiered design approach and was assessed through a constant comparative method of data analysis of interview responses, classroom observations and worksheet-assessments. The results showed that the use of K-W-L Charts influenced the teachers and learners toward a more inquiry-based approach and facilitated a more student-centered and collaborative learning environment, raising the level of interest and the amount of personal input given by the students.

Limbal mesenchymal stromal cells (L-MSC) display immunosuppressive properties across donor and species boundaries

Purpose: We have evaluated the immunosuppressive properties of L-MSC with the view to using these cells in allogeneic cell therapies for corneal disorders. We hypothesized that L-MSC cultures would suppress T-cell activation, in a similar way to those established from human bone marrow (BM-MSC). Methods: MSC cultures were established from the limbal stroma of cadaveric donor eye tissue (up to 1 week postmortem) using either conventional serum-supplemented growth medium or a commercial serum-free medium optimized for bone marrow derived MSC (MesenCult-XF system). The MSC phenotype was examined by flow cytometry according to current and emerging markers for human MSC. Immunosuppressive properties were assessed using a mixed lymphocyte reaction (MLR) assay, whereby the white cell fraction from two immunologically incompatible blood donors are cultured together in direct contact with growth arrested MSC. T-cell activation (proliferation) was measured by uptake of tritiated thymidine. Human L-MSC were tested in parallel with human BM-MSC and rabbit L-MSC. Human and rabbit L-MSC were also tested for their ability to stimulate the growth of limbal epithelial (LE) cells in colony formation assays (for both human as well as rabbit LE cells). Results: L-MSC cultures were >95% negative for CD34, CD45 and HLA-DR and positive for CD73, CD90, CD105 and HLA-ABC. Modest levels (30%) of CD146 expression were observed for L-MSC cultures grown in serum-supplemented growth medium, but not those grown in MesenCult-XF. All MSC cultures derived from both human and rabbit tissue suppressed T-cell activation to varying degrees according to culture technique and species (MesenCult-XF >> serum-fed cultures, rabbit L-MSC >> human L-MSC). All L-MSC stimulated colony formation by LE cells irrespectively of the combination of cell species used. Conclusions: L-MSC display immunosuppressive qualities, in addition to their established non-immunogenic cell surface marker profile, and stimulate LE cell growth in vitro across species boundaries. These results support the potential use of allogeneic or even xenogeneic L-MSC in the treatment of corneal disorders.

Metabolomics coupled with proteomics advancing drug discovery toward more agile development of targeted combination therapies

To enhance the therapeutic efficacy and reduce the adverse effects of traditional Chinese medicine, practitioners often prescribe combinations of plant species and/or minerals, called formulae. Unfortunately, the working mechanisms of most of these compounds are difficult to determine and thus remain unknown. In an attempt to address the benefits of formulae based on current biomedical approaches, we analyzed the components of Yinchenhao Tang, a classical formula that has been shown to be clinically effective for treating hepatic injury syndrome. The three principal components of Yinchenhao Tang are Artemisia annua L., Gardenia jasminoids Ellis, and Rheum Palmatum L., whose major active ingredients are 6,7-dimethylesculetin (D), geniposide (G), and rhein (R), respectively. To determine the mechanisms underlying the efficacy of this formula, we conducted a systematic analysis of the therapeutic effects of the DGR compound using immunohistochemistry, biochemistry, metabolomics, and proteomics. Here, we report that the DGR combination exerts a more robust therapeutic effect than any one or two of the three individual compounds by hitting multiple targets in a rat model of hepatic injury. Thus, DGR synergistically causes intensified dynamic changes in metabolic biomarkers, regulates molecular networks through target proteins, has a synergistic/additive effect, and activates both intrinsic and extrinsic pathways.

A systematic review of mathematical models of mosquito-borne pathogen transmission: 1970-2010

Mathematical models of mosquito-borne pathogen transmission originated in the early twentieth century to provide insights into how to most effectively combat malaria. The foundations of the Ross–Macdonald theory were established by 1970. Since then, there has been a growing interest in reducing the public health burden of mosquito-borne pathogens and an expanding use of models to guide their control. To assess how theory has changed to confront evolving public health challenges, we compiled a bibliography of 325 publications from 1970 through 2010 that included at least one mathematical model of mosquito-borne pathogen transmission and then used a 79-part questionnaire to classify each of 388 associated models according to its biological assumptions. As a composite measure to interpret the multidimensional results of our survey, we assigned a numerical value to each model that measured its similarity to 15 core assumptions of the Ross–Macdonald model. Although the analysis illustrated a growing acknowledgement of geographical, ecological and epidemiological complexities in modelling transmission, most models during the past 40 years closely resemble the Ross–Macdonald model. Modern theory would benefit from an expansion around the concepts of heterogeneous mosquito biting, poorly mixed mosquito-host encounters, spatial heterogeneity and temporal variation in the transmission process.

Career implications of multi-layered institutional settings

Project work has grown significantly in volume and recognition in recent decades as projects have ‘become a common form of work organization in all sectors of the economy’ (Lindgren & Packendorff, 2006: 841). This increase in project-based work is just one of the many changes that have been affecting the nature of work, the employment relationship and the associated conceptualization and experience of careers (Baruch, 2004b; Söderlund & Bredin, 2006). A career can be defined as a process of development along a path of work experience and roles in one or more organizations (Baruch & Rosenstein, 1992), and careers involving project-based work take place within multi layered institutional settings. Projects are generally undertaken by small temporary organizations (Ekstedt, Lundin, Söderholm & Wirdenius, 1999; Pettigrew, 2003; Söderlund, 2012) which in turn may form part of larger, permanent entities; involve people drawn from a number of disciplines and organizations; or be formed as partnerships, joint ventures or strategic alliances between two or more organizations (Scott, 2007).