Ectopic overexpression of a mungbean vacuolar Na+/H+ antiporter gene (VrNHX1) leads to increased salinity stress tolerance in transgenic Vigna unguiculata L. Walp

Salinity is a major threat to sustainable agriculture worldwide. Plant NHX exchangers play an important role in conferring salt tolerance under salinity stress. In this study, a vacuolar Na+/H+ antiporter gene VrNHX1 (Genbank Accession No. JN656211.1) from mungbean (Vigna radiata) was introduced into cowpea (Vigna unguiculata) by the Agrobacterium tumefaciens-mediated transformation method. Polymerase chain reaction and Southern blot hybridization confirmed the stable integration of VrNHX1 into the cowpea genome. Comparative expression analysis by semi-quantitative RT-PCR revealed higher expression of VrNHX1 in transgenic cowpea plants than wild-type. Under salt stress conditions, T2 transgenic 35S:VrNHX1 cowpea lines exhibited higher tolerance to 200 mM NaCl treatment than wild-type. Furthermore, T2 transgenic 35S:VrNHX1 lines maintained a higher K+/Na+ ratio in the aerial parts under salt stress and accumulated higher [Na+] in roots than wild-type. Physiological analysis revealed lower levels of lipid peroxidation, hydrogen peroxide and oxygen radical production but higher levels of relative water content and proline, ascorbate and chlorophyll contents in T2 transgenic 35S:VrNHX1 lines.

A Survey of Viruses Affecting Dry Bean and Cowpea in Lebanon

A survey was conducted to identify viruses affecting dry bean (Phaseolus vulgaris) and cowpea (Vigna unguiculata) in Lebanon. Three hundred and thirty four samples exhibiting virus-like symptoms were collected from 13 different locations during the fall growing season of 1984. Samples were stored at 20 deg C until they were tested by the enzyme-linked immunosorbent assay (ELISA) for the presence of blackeye cowpea mosaic virus (B1CMV), bean yellow mosaic virus (BYMV), bean common mosaic virus (BCMV) and cucumber mosaic virus (CMV). In preliminary tests, the extraction buffer 0.1M phosphate + O.1MEDTA, pH 7.4 was found to be far better than the standard extraction buffer and, accordingly, was used for virus extraction for all field samples. Results obtained indicated that around 50% of the bean samples tested were infected with B1CMV. Incidence of BCMV, BYMV and CMV in the samples tested were 4,4 and 1.7%, respectively. B1CMV was detected in 10 locations, whereas, BYMV, BCMV and CMV were found in 1,4 and 4 locations, respectively. Mixed infections such as BCMV, BICMV, BCMV+CMV, BYMV+CMV and BICMV+BCMV+CMV were detected. In 35% of the samples assayed, the causal virus was not identified

Context-Based Modelling of Interferometric Signals for the Assessment of Tear-Film Suface Quality

Modelling of interferometric signals related to tear film surface quality is considered. In the context of tear film surface quality estimation in normal healthy eyes, two clinical parameters are of interest: the build-up time, and the average interblink surface quality. The former is closely related to the signal derivative while the latter to the signal itself. Polynomial signal models, chosen for a particular set of noisy interferometric measurements, can be optimally selected, in some sense, with a range of information criteria such as AIC, MDL, Cp, and CME. Those criteria, however, do not always guarantee that the true derivative of the signal is accurately represented and they often overestimate it. Here, a practical method for judicious selection of model order in a polynomial fitting to a signal is proposed so that the derivative of the signal is adequately represented. The paper highlights the importance of context-based signal modelling in model order selection.

Compliant cryptologic protocols

Literally, the word compliance suggests conformity in fulfilling official requirements. The thesis presents the results of the analysis and design of a class of protocols called compliant cryptologic protocols (CCP). The thesis presents a notion for compliance in cryptosystems that is conducive as a cryptologic goal. CCP are employed in security systems used by at least two mutually mistrusting sets of entities. The individuals in the sets of entities only trust the design of the security system and any trusted third party the security system may include. Such a security system can be thought of as a broker between the mistrusting sets of entities. In order to provide confidence in operation for the mistrusting sets of entities, CCP must provide compliance verification mechanisms. These mechanisms are employed either by all the entities or a set of authorised entities in the system to verify the compliance of the behaviour of various participating entities with the rules of the system. It is often stated that confidentiality, integrity and authentication are the primary interests of cryptology. It is evident from the literature that authentication mechanisms employ confidentiality and integrity services to achieve their goal. Therefore, the fundamental services that any cryptographic algorithm may provide are confidentiality and integrity only. Since controlling the behaviour of the entities is not a feasible cryptologic goal,the verification of the confidentiality of any data is a futile cryptologic exercise. For example, there exists no cryptologic mechanism that would prevent an entity from willingly or unwillingly exposing its private key corresponding to a certified public key. The confidentiality of the data can only be assumed. Therefore, any verification in cryptologic protocols must take the form of integrity verification mechanisms. Thus, compliance verification must take the form of integrity verification in cryptologic protocols. A definition of compliance that is conducive as a cryptologic goal is presented as a guarantee on the confidentiality and integrity services. The definitions are employed to provide a classification mechanism for various message formats in a cryptologic protocol. The classification assists in the characterisation of protocols, which assists in providing a focus for the goals of the research. The resulting concrete goal of the research is the study of those protocols that employ message formats to provide restricted confidentiality and universal integrity services to selected data. The thesis proposes an informal technique to understand, analyse and synthesise the integrity goals of a protocol system. The thesis contains a study of key recovery,electronic cash, peer-review, electronic auction, and electronic voting protocols. All these protocols contain message format that provide restricted confidentiality and universal integrity services to selected data. The study of key recovery systems aims to achieve robust key recovery relying only on the certification procedure and without the need for tamper-resistant system modules. The result of this study is a new technique for the design of key recovery systems called hybrid key escrow. The thesis identifies a class of compliant cryptologic protocols called secure selection protocols (SSP). The uniqueness of this class of protocols is the similarity in the goals of the member protocols, namely peer-review, electronic auction and electronic voting. The problem statement describing the goals of these protocols contain a tuple,(I, D), where I usually refers to an identity of a participant and D usually refers to the data selected by the participant. SSP are interested in providing confidentiality service to the tuple for hiding the relationship between I and D, and integrity service to the tuple after its formation to prevent the modification of the tuple. The thesis provides a schema to solve the instances of SSP by employing the electronic cash technology. The thesis makes a distinction between electronic cash technology and electronic payment technology. It will treat electronic cash technology to be a certification mechanism that allows the participants to obtain a certificate on their public key, without revealing the certificate or the public key to the certifier. The thesis abstracts the certificate and the public key as the data structure called anonymous token. It proposes design schemes for the peer-review, e-auction and e-voting protocols by employing the schema with the anonymous token abstraction. The thesis concludes by providing a variety of problem statements for future research that would further enrich the literature.

Good practice for enhancing the engagement and success of commencing students

There is widespread recognition that higher education institutions (HEIs) must actively support commencing students to ensure equity in access to the opportunities afforded by higher education. This role is particularly critical for students who because of educational, cultural or financial disadvantage or because they are members of social groups currently under-represented in higher education, may require additional transitional support to “level the playing field.” The challenge faced by HEIs is to provide this “support” in a way that is integrated into regular teaching and learning practices and reaches all commencing students. The Student Success Program (SSP) is an intervention in operation at the Queensland University of Technology (QUT) designed to identify and support those students deemed to be at risk of disengaging from their learning and their institution. Two sets of evidence of the impact of the SSP are presented: First, its expansion (a) from a one-faculty pilot project (Nelson, Duncan & Clarke, 2009) to all faculties and (b) into a variety of applications mirroring the student life cycle; and second, an evaluation of the impact of the SSP on students exposed to it. The outcomes suggest that: the SSP is an example of good practice that can be successfully applied to a variety of learning contexts and student enrolment situations; and the impact of the intervention on student persistence is sustained for at least 12 months and positively influences student retention. It is claimed that the good practice evidenced by the SSP is dependent on its integration into the broader First Year Experience Program at QUT as an example of transition pedagogy in action.

Start Smart : QUT’s trial support program for commencing undergraduate students who have not completed year 12 in the past two years

Commencing students in undergraduate degrees who identify as mature age students experience particular issues when faced with enrolment into university as an adult learner (Bird & Morgan, 2003). In line with QUT’s commitment to “supporting all commencing students to adjust successfully to study at QUT by providing a strong transition experience” (QUT, 2008, 6.2.1), the Start Smart trial program was developed and implemented for Semester 1, 2012. The Start Smart trial program consists of an orientation event, wrapped around and supported by existing First Year Experience (FYE) and Retention strategies within QUT, namely the Student Success Program (SSP) and the Peer Programs Strategy (PPS). This report examines the motivations for designing a program as a response to the needs of a cohort that are unique amongst all commencing undergraduate students. Participants will be asked to consider the implications of delivering special and unique orientation events to specific cohorts, and the long term sustainability of such programs within their own university structures.

The evolution of QUT’s Student Success Program : 20,000 students later

“The Student Success Program (SSP) is a monitoring and early intervention program in operation at QUT designed to identify and support those students deemed to be at risk of disengaging for their learning and their institution” (Nelson, Quinn, Marrington & Clarke, 2011, p. 83). This report reflects on the development of the program since its inception in 2007. In acknowledging similar initiatives within the sector that monitor student learning engagement, the Nuts & Bolts session allows for identification and discussion of the critical success factors for these intervention and support programs.

Social signal processing for pain monitoring using a hidden conditional random field

utomatic pain monitoring has the potential to greatly improve patient diagnosis and outcomes by providing a continuous objective measure. One of the most promising methods is to do this via automatically detecting facial expressions. However, current approaches have failed due to their inability to: 1) integrate the rigid and non-rigid head motion into a single feature representation, and 2) incorporate the salient temporal patterns into the classification stage. In this paper, we tackle the first problem by developing a “histogram of facial action units” representation using Active Appearance Model (AAM) face features, and then utilize a Hidden Conditional Random Field (HCRF) to overcome the second issue. We show that both of these methods improve the performance on the task of pain detection in sequence level compared to current state-of-the-art-methods on the UNBC-McMaster Shoulder Pain Archive.

Design, implementation and multisite evaluation of a system suitability protocol for the quantitative assessment of instrument performance in liquid chromatography-multiple reaction monitoring-MS (LC-MRM-MS)

Multiple reaction monitoring (MRM) mass spectrometry coupled with stable isotope dilution (SID) and liquid chromatography (LC) is increasingly used in biological and clinical studies for precise and reproducible quantification of peptides and proteins in complex sample matrices. Robust LC-SID-MRM-MS-based assays that can be replicated across laboratories and ultimately in clinical laboratory settings require standardized protocols to demonstrate that the analysis platforms are performing adequately. We developed a system suitability protocol (SSP), which employs a predigested mixture of six proteins, to facilitate performance evaluation of LC-SID-MRM-MS instrument platforms, configured with nanoflow-LC systems interfaced to triple quadrupole mass spectrometers. The SSP was designed for use with low multiplex analyses as well as high multiplex approaches when software-driven scheduling of data acquisition is required. Performance was assessed by monitoring of a range of chromatographic and mass spectrometric metrics including peak width, chromatographic resolution, peak capacity, and the variability in peak area and analyte retention time (RT) stability. The SSP, which was evaluated in 11 laboratories on a total of 15 different instruments, enabled early diagnoses of LC and MS anomalies that indicated suboptimal LC-MRM-MS performance. The observed range in variation of each of the metrics scrutinized serves to define the criteria for optimized LC-SID-MRM-MS platforms for routine use, with pass/fail criteria for system suitability performance measures defined as peak area coefficient of variation <0.15, peak width coefficient of variation <0.15, standard deviation of RT <0.15 min (9 s), and the RT drift <0.5min (30 s). The deleterious effect of a marginally performing LC-SID-MRM-MS system on the limit of quantification (LOQ) in targeted quantitative assays illustrates the use and need for a SSP to establish robust and reliable system performance. Use of a SSP helps to ensure that analyte quantification measurements can be replicated with good precision within and across multiple laboratories and should facilitate more widespread use of MRM-MS technology by the basic biomedical and clinical laboratory research communities.

The Enamovirus P0 protein is a silencing suppressor which inhibits local and systemic RNA silencing through AGO1 degradation

The P0 protein of poleroviruses and P1 protein of sobemoviruses suppress the plant's RNA silencing machinery. Here we identified a silencing suppressor protein (SSP), P0PE, in the Enamovirus Pea enation mosaic virus-1 (PEMV-1) and showed that it and the P0s of poleroviruses Potato leaf roll virus and Cereal yellow dwarf virus have strong local and systemic SSP activity, while the P1 of Sobemovirus Southern bean mosaic virus supresses systemic silencing. The nuclear localized P0PE has no discernable sequence conservation with known SSPs, but proved to be a strong suppressor of local silencing and a moderate suppressor of systemic silencing. Like the P0s from poleroviruses, P0PE destabilizes AGO1 and this action is mediated by an F-box-like domain. Therefore, despite the lack of any sequence similarity, the poleroviral and enamoviral SSPs have a conserved mode of action upon the RNA silencing machinery. © 2012 Elsevier Inc.

Linking ecological condition and the soundscape in fragmented Australian forests

Natural landscapes are increasingly subjected to anthropogenic pressure and fragmentation resulting in reduced ecological condition. In this study we examined the relationship between ecological condition and the soundscape in fragmented forest remnants of south-east Queensland, Australia. The region is noted for its high biodiversity value and increased pressure associated with habitat fragmentation and urbanisation. Ten sites defined by a distinct open eucalypt forest community dominated by spotted gum (Corymbia citriodora ssp. variegata) were stratified based on patch size and patch connectivity. Each site underwent a series of detailed vegetation condition and landscape assessments, together with bird surveys and acoustic analysis using relative soundscape power. Univariate and multivariate analyses indicated that the measurement of relative soundscape power reflects ecological condition and bird species richness, and is dependent on the extent of landscape fragmentation. We conclude that acoustic monitoring technologies provide a cost effective tool for measuring ecological condition, especially in conjunction with established field observations and recordings.

Large-scale interlaboratory study to develop, analytically validate and apply highly multiplexed, quantitative peptide assays to measure cancer-relevant proteins in plasma

There is an increasing need in biology and clinical medicine to robustly and reliably measure tens-to-hundreds of peptides and proteins in clinical and biological samples with high sensitivity, specificity, reproducibility and repeatability. Previously, we demonstrated that LC-MRM-MS with isotope dilution has suitable performance for quantitative measurements of small numbers of relatively abundant proteins in human plasma, and that the resulting assays can be transferred across laboratories while maintaining high reproducibility and quantitative precision. Here we significantly extend that earlier work, demonstrating that 11 laboratories using 14 LC-MS systems can develop, determine analytical figures of merit, and apply highly multiplexed MRM-MS assays targeting 125 peptides derived from 27 cancer-relevant proteins and 7 control proteins to precisely and reproducibly measure the analytes in human plasma. To ensure consistent generation of high quality data we incorporated a system suitability protocol (SSP) into our experimental design. The SSP enabled real-time monitoring of LC-MRM-MS performance during assay development and implementation, facilitating early detection and correction of chromatographic and instrumental problems. Low to sub-nanogram/mL sensitivity for proteins in plasma was achieved by one-step immunoaffinity depletion of 14 abundant plasma proteins prior to analysis. Median intra- and inter-laboratory reproducibility was <20%, sufficient for most biological studies and candidate protein biomarker verification. Digestion recovery of peptides was assessed and quantitative accuracy improved using heavy isotope labeled versions of the proteins as internal standards. Using the highly multiplexed assay, participating laboratories were able to precisely and reproducibly determine the levels of a series of analytes in blinded samples used to simulate an inter-laboratory clinical study of patient samples. Our study further establishes that LC-MRM-MS using stable isotope dilution, with appropriate attention to analytical validation and appropriate quality c`ontrol measures, enables sensitive, specific, reproducible and quantitative measurements of proteins and peptides in complex biological matrices such as plasma.

Genetic testing for haemochromatosis in patients with chondrocalcinosis

Hereditary haemochromatosis (HH) is the most common lethal monogenic human disease, affecting roughly 1 in 300 white northern Europeans. Homozygosity for the C282Y polymorphism within the HFE gene causes more than 80% of cases, with compound heterozygosity of the C282Y and H63D polymorphism also increasing susceptibility to disease. The aim of this study was to determine the frequency of the C282Y and H63D polymorphisms in the disease, and to assess the risk of HH in heterozygotes for the C282Y polymorphism. 128 patients were recruited because of either radiographic chondrocalcinosis (at least bicompartmental knee disease or joints other than the knee involved) or CPPD pseudogout. Genotyping of the HFE C282Y and H63D mutations was performed using PCR/SSP and genotypes for the C282Y polymorphism confirmed by PCR/RFLP. Historical white European control data were used for comparison. Two previously undiagnosed C282Y homozygotes (1.6%), and 16 C282Y heterozygotes (12.5%), including four (3.1%) C282Y/ H63D compound heterozygotes were identified. This represents a significant overrepresentation of C282Y homozygotes (relative risk 3.4, p-0.037), but the number of heterozygotes was not significantly increased. At a cost per test of £1 for each subject, screening all patients with chondrocalcinosis using the above ascertainment criteria costs only £64 for each case of haemochromatosis identified, clearly a highly cost effective test given the early mortality associated with untreated haemochromatosis. Routine screening for haemochromatosis in patients with appreciable chondrocatcinosis is recommended.