A three-nucleotide mutation altering the Maize streak virus Rep pRBR-interaction motif reduces symptom severity in maize and partially reverts at high frequency without restoring pRBR-Rep binding

Geminivirus infectivity is thought to depend on interactions between the virus replication-associated proteins Rep or RepA and host retinoblastoma-related proteins (pRBR), which control cell-cycle progression. It was determined that the substitution of two amino acids in the Maize streak virus (MSV) RepA pRBR-interaction motif (LLCNE to LLCLK) abolished detectable RepA-pRBR interaction in yeast without abolishing infectivity in maize. Although the mutant virus was infectious in maize, it induced less severe symptoms than the wild-type virus. Sequence analysis of progeny viral DNA isolated from infected maize enabled detection of a high-frequency single-nucleotide reversion of C(601)A in the 3 nt mutated sequence of the Rep gene. Although it did not restore RepA-pRBR interaction in yeast, sequence-specific PCR showed that, in five out of eight plants, the C(601)A reversion appeared by day 10 post-inoculation. In all plants, the C(601)A revertant eventually completely replaced the original mutant population, indicating a high selection pressure for the single-nucleotide reversion. Apart from potentially revealing an alternative or possibly additional function for the stretch of DNA that encodes the apparently non-essential pRBR-interaction motif of MSV Rep, the consistent emergence and eventual dominance of the C(601)A revertant population might provide a useful tool for investigating aspects of MSV biology, such as replication, mutation and evolution rates, and complex population phenomena, such as competition between quasispecies and population turnover. © 2005 SGM.

High level protein expression in plants through the use of a novel autonomously replicating geminivirus shuttle vector

We constructed a novel autonomously replicating gene expression shuttle vector, with the aim of developing a system for transiently expressing proteins at levels useful for commercial production of vaccines and other proteins in plants. The vector, pRIC, is based on the mild strain of the geminivirus Bean yellow dwarf virus (BeYDV-m) and is replicationally released into plant cells from a recombinant Agrobacterium tumefaciens Ti plasmid. pRIC differs from most other geminivirus-based vectors in that the BeYDV replication-associated elements were included in cis rather than from a co-transfected plasmid, while the BeYDV capsid protein (CP) and movement protein (MP) genes were replaced by an antigen encoding transgene expression cassette derived from the non-replicating A. tumefaciens vector, pTRAc. We tested vector efficacy in Nicotiana benthamiana by comparing transient cytoplasmic expression between pRIC and pTRAc constructs encoding either enhanced green fluorescent protein (EGFP) or the subunit vaccine antigens, human papillomavirus subtype 16 (HPV-16) major CP L1 and human immunodeficiency virus subtype C p24 antigen. The pRIC constructs were amplified in planta by up to two orders of magnitude by replication, while 50% more HPV-16 L1 and three- to seven-fold more EGFP and HIV-1 p24 were expressed from pRIC than from pTRAc. Vector replication was shown to be correlated with increased protein expression. We anticipate that this new high-yielding plant expression vector will contribute towards the development of a viable plant production platform for vaccine candidates and other pharmaceuticals. © 2009 Blackwell Publishing Ltd.

Reaching high-risk adolescents : a process evaluation of a school based injury prevention program

High-risk adolescents are a population most vulnerable to harm from injury due to increased engagement in risk taking behaviour. There is a gap in the literature regarding how universal school based injury prevention programs apply to high-risk adolescents. This study involves a component of the process evaluation of a school based injury prevention program, as it relates to high-risk adolescents (13-14 years)...

Accuracy of FEM and BEM for electromagnetic flux calculations in high power applications

The development and design of electric high power devices with electromagnetic computer-aided engineering (EM-CAE) software such as the Finite Element Method (FEM) and Boundary Element Method (BEM) has been widely adopted. This paper presents the analysis of a Fault Current Limiter (FCL), which acts as a high-voltage surge protector for power grids. A prototype FCL was built. The magnetic flux in the core and the resulting electromagnetic forces in the winding of the FCL were analyzed using both FEM and BEM. An experiment on the prototype was conducted in a laboratory. The data obtained from the experiment is compared to the numerical solutions to determine the suitability and accuracy of the two methods.

Developmental components of resting ventilation among high- and low-altitude Andean children and adults

This paper evaluates the age-associated changes of resting ventilation of 115 high- and low-altitude Aymara subjects, of whom 61 were from the rural Aymara village of Ventilla situated at an average altitude of 4,200 m and 54 from the rural village of Caranavi situated at an average altitude of 900 m. Comparison of the age patterns of resting ventilation suggests the following conclusions: 1) the resting ventilation (ml/kg/min) of high-altitude natives is markedly higher than that of low-altitude natives; 2) the age decline of ventilation is similar in both lowlanders and highlanders, but the starting point and therefore the age decline are much higher at high altitude; 3) the resting ventilation that characterizes high-altitude Andean natives is developmentally expressed in the same manner as it is at low altitude; and 4) the resting ventilation (ml/kg/min) of Aymara high-altitude natives is between 40–80% lower than that of Tibetans. Am J Phys Anthropol 109:295–301, 1999. © 1999 Wiley-Liss, Inc.

The relationship between fundamental movement skills and self-reported physical activity during Finnish junior high school

Background: Previous studies have shown that fundamental movement skills (FMS) and physical activity are related. Specifically, earlier studies have demonstrated that the ability to perform a variety of FMS increases the likelihood of children participating in a range of physical activities throughout their lives. To date, however, there have not been studies focused on the development of, or the relationship between, these variables through junior high school (that is, between the ages of 13 and 15). Such studies might provide important insights into the relationships between FMS and physical activity during adolescence, and suggest ways to design more effective physical education programmes for adolescents. Purpose: The main purposes of the study are: (1) to investigate the development of the students' self-reported physical activity and FMS from Grade 7 to Grade 9, (2) to analyse the associations among the students' FMS and self-reported physical activity through junior high school, (3) to analyse whether there are gender differences in research tasks one and/or two. Participants and setting: The participants in the study were 152 Finnish students, aged 13 and enrolled in Grade 7 at the commencement of the study. The sample included 66 girls and 86 boys who were drawn from three junior high schools in Middle Finland. Research design and data collection: Both the FMS tests and questionnaires pertaining to self-reported physical activity were completed annually during a 3 year period: in August (when the participants were in Grade 7), January (Grade 8), and in May (Grade 9). Data analysis: Repeated measures multivariate analysis of variances (MANOVAs) were used to analyse the interaction between gender and time (three measurement points) in FMS test sumscores and self-reported physical activity scores. The relationships between self-reported physical activity scores and fundamental movement skill sumscores through junior high school were analysed using Structural Equation Modelling (SEM) with LISREL 8.80 software. Findings: The MANOVA for self-reported physical activity demonstrated that both genders' physical activity decreased through junior high school. The MANOVA for the FMS revealed that the boys' FMS sumscore increased whereas the girls' skills decreased through junior high school. The SEM and squared multiple correlations revealed FMS in Grades 7 and 8 as well as physical activity in Grade 9 explained FMS in Grade 9. The portion of prediction was 69% for the girls and 55% for the boys. Additionally, physical activity measured in Grade 7 and FMS measured in Grade 9 explained physical activity in Grade 9. The portion of prediction was 12% for the girls and 29% for the boys. In the boys' group, three additional paths were found; FMS in Grade 7 explained physical activity in Grade 9, physical activity in Grade 7 explained FMS in Grade 8, and physical activity in Grade 7 explained physical activity in Grade 8. Conclusions: The study suggests that supporting and encouraging FMS and physical activity are co-related and when considering combined scores there is a greater likelihood of healthy lifelong outcomes. Therefore, the conclusion can be drawn that FMS curriculum in school-based PE is a plausible way to ensure good lifelong outcomes. Earlier studies support that school physical education plays an important role in developing students FMS and is in a position to thwart the typical decline of physical activity in adolescence. These concepts are particularly important for adolescent girls as this group reflects the greatest decline in physical activity during the adolescent period.

Reduced expression of retinoblastoma gene product (pRB) and high expression of p53 are associated with poor prognosis in ovarian cancer

Paraffin sections (n = 168, 27 benign, 16 low malignant potential [LMP] and 125 malignant tumours) from epithelial ovarian tumours were evaluated immunohistochemically for expression of retinoblastoma gene product (pRB) and p53 protein, and the relationship among pRB, p53 and cyclin-dependent kinase inhibitor 2 (CDKN2) gene product p16INK4A (p16) was analysed, following our previous study of p16. Forty-one percent of the benign, 50% of the LMP and most (71%) of the malignant tumours showed high pRB expression. High expression of pRB (>50% pRB-positive cells) significantly correlated with non-mucinous histological subtypes. Reduced pRB expression, substage and residual disease were significant predictors for poor prognosis in stage I patients. All the benign and most of the LMP (81%) tumours were in either the p53-negative or low p53-positive category, but nearly half of the malignant tumours had high p53 expression. High p53 accumulation was found in non-mucinous, high grade and late stage tumours. For well-differentiated carcinomas, high p53 expression was a predictor of poor prognosis. However, even though high p53 expression was not associated with histological subtype, stage or the presence of residual disease, high p53 expression was not an independent predictor when all clinical parameters were combined. For all ovarian cancers, a close correlation was found between high p53 and high p16 expression. The relationship between the expression of pRB and p16 depended on tumour stage. In stage I tumours, high pRB was associated with low p16 reactivity. On the other hand, most advanced tumours showed both high pRB and high p16 reactivity. Int. J. Cancer 74:407–415, 1997. © 1997 Wiley-Liss, Inc.

Full compensation no longer sacrosanct : reflections on the past and future economic loss ‘cap’ for high income earners

The civil liability provisions relating to the assessment of damages for past and future economic loss have abrogated the common law principle of full compensation by imposing restrictions on the damages award, most commonly by a “three times average weekly earnings” cap. This consideration of the impact of those provisions is informed by a case study of the Supreme Court of Victoria Court of Appeal decision, Tuohey v Freemasons Hospital (Tuohey) , which addressed the construction and arithmetic operation of the Victorian cap for high income earners. While conclusions as to operation of the cap outside of Victoria can be drawn from Tuohey, a number of issues await judicial determination. These issues, which include the impact of the damages caps on the calculation of damages for economic loss in the circumstances of fluctuating income; vicissitudes; contributory negligence; claims per quod servitum amisit; and claims by dependants, are identified and potential resolutions discussed.