The role of the yeast cleavage and polyadenylation factor subunit Ydh1p/Cft2p in pre-mRNA 3'-end formation

Cleavage and polyadenylation factor (CPF) is a multi‐protein complex that functions in pre‐mRNA 3′‐end formation and in the RNA polymerase II (RNAP II) transcription cycle. Ydh1p/Cft2p is an essential component of CPF but its precise role in 3′‐end processing remained unclear. We found that mutations in YDH1 inhibited both the cleavage and the polyadenylation steps of the 3′‐end formation reaction in vitro. Recently, we demonstrated that an important function of CPF lies in the recognition of poly(A) site sequences and RNA binding analyses suggesting that Ydh1p/Cft2p interacts with the poly(A) site region. Here we show that mutant ydh1 strains are deficient in the recognition of the ACT1 cleavage site in vivo. The C‐terminal domain (CTD) of RNAP II plays a major role in coupling 3′‐end processing and transcription. We provide evidence that Ydh1p/Cft2p interacts with the CTD of RNAP II, several other subunits of CPF and with Pcf11p, a component of CF IA. We propose that Ydh1p/Cft2p contributes to the formation of important interaction surfaces that mediate the dynamic association of CPF with RNAP II, the recognition of poly(A) site sequences and the assembly of the polyadenylation machinery on the RNA substrate.

Independent functions of yeast Pcf11p in pre-mRNA 3' end processing and in transcription termination

Pcf11p, an essential subunit of the yeast cleavage factor IA, is required for pre‐mRNA 3′ end processing, binds to the C‐terminal domain (CTD) of the largest subunit of RNA polymerase II (RNAP II) and is involved in transcription termination. We show that the conserved CTD interaction domain (CID) of Pcf11p is essential for cell viability. Interestingly, the CTD binding and 3′ end processing activities of Pcf11p can be functionally uncoupled from each other and provided by distinct Pcf11p fragments in trans. Impaired CTD binding did not affect the 3′ end processing activity of Pcf11p and a deficiency of Pcf11p in 3′ end processing did not prevent CTD binding. Transcriptional run‐on analysis with the CYC1 gene revealed that loss of cleavage activity did not correlate with a defect in transcription termination, whereas loss of CTD binding did. We conclude that Pcf11p is a bifunctional protein and that transcript cleavage is not an obligatory step prior to RNAP II termination.

Determining gas sampling timelines for estimating emissions in small chamber incubation experiments

A laboratory experiment was set up in small chambers for monitoring greenhouse gas emissions and determining the most suitable time for sampling. A six-treatment experiment was conducted, including a one week pre-incubation and a week for incubation. Timelines for sampling were 1, 2, 3, 6 and 24 hours after closing the lid of the incubation chambers. Variation in greenhouse gas fluxes was high due to the time of sampling. The rates of gas emissions increased in first three hours and decreased afterward. The rates of greenhouse gas emissions at 3 hours after closing lids was close to the mean for the 24-h period.

Emotional climate and behavioral management during sleep time in early childhood education settings

The majority of children cease napping between 3 and 5 years of age yet, internationally, the allocation of a sleep time during the day for children of this age remains a practice in many early childhood education (ECE) settings. These dual circumstances present a disjuncture between children's sleep needs and center practices, that may cause conflict for staff, increase stress for children and escalate negative emotional climate in the room. Testing this hypothesis requires observation of both the emotional climate and behavioral management used in ECE rooms that extends into the sleep time. This study was the first to apply the Classroom Assessment and Scoring System (CLASS) Pre-K (Pianta, La Paro, & Hamre, 2008) to observe the emotional climate and behavioral management during sleep time. Pilot results indicated that the CLASS Pre-K functioned reliably to measure emotional climate and behavioral management in sleep time. However, new sleep-specific examples of the dimensions used were developed, to help orient fieldworkers to the CLASS Pre-K rating system in the sleep time context. The CLASS was then used to assess emotional climate and behavior management between the non-sleep and sleep time sessions, in 113 ECE rooms in Queensland, Australia. In these rooms 2.114 children were observed. Of these children, 71% did not sleep at any point during the allotted sleep times. There was a significant drop in emotional climate and behavioral management between the non-sleep and sleep-time sessions. Furthermore, the duration of mandated sleep time (a period of time where no activities are provided to non-sleeping children) accounted for significant independent variance in the observed emotional climate during sleep-time. The CLASS Pre-K presents a valuable tool to assess the emotional climate and behavior management during sleep-time and draws attention to the need for further studies of sleep time in ECE settings.

Alcohol-related emergency department injury presentations in Queensland adolescents and young adults over a 13-year period

Introduction and Aims. The rate of alcohol-related emergency department (ED) presentations in young people has increased dramatically in recent decades. Injuries are the most common type of youth alcohol-related ED presentation, yet little is known about these injuries in young people. This paper describes the characteristics of alcohol-related ED injury presentations in young people over a 13-year period and determines if they differ by gender and/or age group (adolescents: 12–17 years; young adults: 18–24 years). Design and Method. The Queensland Injury Surveillance Unit (QISU) database collects injury surveillance data at triage in participating EDs throughout Queensland, Australia. A total of 4667 cases of alcohol-related injuries in young people (aged 12–24 years) were identified in the QISU database between January 1999 and December 2011, using an injury surveillance code and nursing triage text-based search strategy. Results. Overall, young people accounted for 38% of all QISU alcohol-related ED injury presentations in patients aged 12 years or over. The majority of young adults presented with injuries due to violence and falls, whereas adolescents presented due to self-harm or intoxication without other injury. Males presented with injuries due to violence, whereas females presented with alcohol-related self-harm and intoxication. Discussion and Conclusions. There is a need for more effective ways of identifying the degree of alcohol involvement in injuries among young people presenting to EDs.

Changes in outdoor workers' sun-related attitudes, beliefs, and behaviors : a pre-post workplace intervention

OBJECTIVE To evaluate changes in outdoor workers' sun-related attitudes, beliefs, and behaviors in response to a health promotion intervention using a participatory action research process. METHODS Fourteen workplaces across four outdoor industry types worked collaboratively with the project team to develop tailored sun protection action plans. Workers were assessed before and after the 18-month intervention. RESULTS Outdoor workers reported increases in workplace support for sun protection (P < 0.01) and personal use of sun protection (P < 0.01). More workers reported seeking natural shade (+20%) and wearing more personal protective equipment, including broad-brimmed hats (+25%), long-sleeved collared shirts (+19%), and long trousers (+16%). The proportion of workers reporting sunburn over the past 12 months was lower at postintervention (-14%) (P = 0.03); however, the intensity of reported sunburn increased. CONCLUSIONS This intervention was successful in increasing workers' sun protective attitudes, beliefs, and behaviors.

Incidence of paediatric fatal and non-fatal low speed vehicle run over events in Queensland, Australia : eleven year analysis

Background The purpose of this study was to estimate the incidence of fatal and non-fatal Low Speed Vehicle Run Over (LSVRO) events among children aged 0–15 years in Queensland, Australia, at a population level. Methods Fatal and non-fatal LSVRO events that occurred in children resident in Queensland over eleven calendar years (1999-2009) were identified using ICD codes, text description, word searches and medical notes clarification, obtained from five health related data bases across the continuum of care (pre-hospital to fatality). Data were manually linked. Population data provided by the Australian Bureau of Statistics were used to calculate crude incidence rates for fatal and non-fatal LSVRO events. Results There were 1611 LSVROs between 1999–2009 (IR = 16.87/100,000/annum). Incidence of non-fatal events (IR = 16.60/100,000/annum) was 61.5 times higher than fatal events (IR = 0.27/100,000/annum). LSVRO events were more common in boys (IR = 20.97/100,000/annum) than girls (IR = 12.55/100,000/annum), and among younger children aged 0–4 years (IR = 21.45/100000/annum; 39% or all events) than older children (5–9 years: IR = 16.47/100,000/annum; 10–15 years IR = 13.59/100,000/annum). A total of 896 (56.8%) children were admitted to hospital for 24 hours of more following an LSVRO event (IR = 9.38/100,000/annum). Total LSVROs increased from 1999 (IR = 14.79/100,000) to 2009 (IR = 18.56/100,000), but not significantly. Over the 11 year period, there was a slight (non –significant) increase in fatalities (IR = 0.37-0.42/100,000/annum); a significant decrease in admissions (IR = 12.39–5.36/100,000/annum), and significant increase in non-admissions (IR = 2.02-12.77/100,000/annum). Trends over time differed by age, gender and severity. Conclusion This is the most comprehensive, population-based epidemiological study on fatal and non-fatal LSVRO events to date. Results from this study indicate that LSVROs incur a substantial burden. Further research is required on the characteristics and risk factors associated with these events, in order to adequately inform injury prevention. Strategies are urgently required in order to prevent these events, especially among young children aged 0-4 years.

Effects of selective oestrogen receptor modulators on proliferation in tissue cultures of pre- and postmenopausal human endometrium

We characterised the effects of selective oestrogen receptor modulators (SERM) in explant cultures of human endometrium tissue. Endometrium tissues were cultured for 24 h in Millicell-CM culture inserts in serum-free medium in the presence of vehicle,17 beta-estradiol (17 beta-E2,1 nM), oestrogen receptor (ER) antagonist ICI 164.384 (40 nM), and 4-OH-tamoxifen (40 nM), raloxifene (4 nM), lasofoxifene (4 nM)and acolbifene (4 nM). Protein expression of ER alpha, ER beta 1 and Ki-67 were evaluated by immunohistochemistry (IHC). The proliferative fraction was assessed by counting the number of Ki-67 positive cells. Nuclear staining of ER( and ER(1 was observed in the glandular epithelium and stroma of pre- and postmenopausal endometrium. ER(1 protein was also localized in the endothelial cells of blood vessels. Treating premenopausal endometrium tissue with 17 beta-E2 increased the fraction of Ki-67 positive cells (p < 0.001) by 55% in glands compared to the control. Raloxifene (4 nM) increased (p < 0.05) the Ki-67 positive fraction. All other SERMS did not affect proliferation in this model. Treating postmenopausal endometrium with 17(-E2 increased (p < 0.001) the fraction of Ki-67 positive cells by 250% in glands compared to the control. A similar effect was also seen for 4-OH-tamoxifen, whereas the rest of SERMs did not stimulate proliferation. We demonstrated that oestradiol increases the fraction of proliferating cells in short term explant cultures of postmenopausal endometrium. In addition, we were able to reveal the agonistic properties of 4-OH-tamoxifen and confirm that raloxifene and next-generation SERMs acolbifene and lasofoxifene were neutral on the human postmenopausal endometrium. (C) 2008 Elsevier Ltd. All rights reserved.

Reproducibility of subjective appetite ratings and ad libitum test meal energy intake in overweight and obese males

Background To determine whether changes in appetite and energy intake (EI) can be detected and play a role in the effectiveness of interventions, it is necessary to identify their variability under normal conditions. We assessed the reproducibility of subjective appetite ratings and ad libitum test meal EI after a standardised pre-load in overweight and obese males. Methods Fifteen overweight and obese males (BMI 30.3 ± 4.9 kg/m2, aged 34.9 ± 10.6 years) completed two identical test days, 7 days apart. Participants were provided with a standardised fixed breakfast (1676 kJ) and 5 h later an ad libitum pasta lunch. An electronic appetite rating system was used to assess subjective ratings before and after the fixed breakfast, and periodically during the postprandial period. EI was assessed at the ad libitum lunch meal. Sample size estimates for paired design studies were calculated. Results Appetite ratings demonstrated a consistent oscillating pattern between test days, and were more reproducible for mean postprandial than fasting ratings. The correlation between ad libitum EI on the two test days was r = 0.78 (P < 0.01). Using a paired design and a power of 0.8, a minimum of 12 participants would be needed to detect a 10 mm change in 5 h postprandial mean ratings and 17 to detect a 500 kJ difference in ad libitum EI. Conclusion Intra-individual variability of appetite and ad libitum test meal EI in overweight and obese males is comparable to previous reports in normal weight adults. Sample size requirements for studies vary depending on the parameter of interest and sensitivity needed.

Supine to standing Cobb angle change in idiopathic scoliosis : the effect of endplate pre-selection

Background Supine imaging modalities provide valuable 3D information on scoliotic anatomy, but the altered spine geometry between the supine and standing positions affects the Cobb angle measurement. Previous studies report a mean 7°-10° Cobb angle increase from supine to standing, but none have reported the effect of endplate pre-selection or whether other parameters affect this Cobb angle difference. Methods Cobb angles from existing coronal radiographs were compared to those on existing low-dose CT scans taken within three months of the reference radiograph for a group of females with adolescent idiopathic scoliosis. Reformatted coronal CT images were used to measure supine Cobb angles with and without endplate pre-selection (end-plates selected from the radiographs) by two observers on three separate occasions. Inter and intra-observer measurement variability were assessed. Multi-linear regression was used to investigate whether there was a relationship between supine to standing Cobb angle change and eight variables: patient age, mass, standing Cobb angle, Risser sign, ligament laxity, Lenke type, fulcrum flexibility and time delay between radiograph and CT scan. Results Fifty-two patients with right thoracic Lenke Type 1 curves and mean age 14.6 years (SD 1.8) were included. The mean Cobb angle on standing radiographs was 51.9° (SD 6.7). The mean Cobb angle on supine CT images without pre-selection of endplates was 41.1° (SD 6.4). The mean Cobb angle on supine CT images with endplate pre-selection was 40.5° (SD 6.6). Pre-selecting vertebral endplates increased the mean Cobb change by 0.6° (SD 2.3, range −9° to 6°). When free to do so, observers chose different levels for the end vertebrae in 39% of cases. Multi-linear regression revealed a statistically significant relationship between supine to standing Cobb change and fulcrum flexibility (p = 0.001), age (p = 0.027) and standing Cobb angle (p < 0.001). The 95% confidence intervals for intra-observer and inter-observer measurement variability were 3.1° and 3.6°, respectively. Conclusions Pre-selecting vertebral endplates causes minor changes to the mean supine to standing Cobb change. There is a statistically significant relationship between supine to standing Cobb change and fulcrum flexibility such that this difference can be considered a potential alternative measure of spinal flexibility.

The effect of previous hamstring strain injuries on the change in eccentric hamstring strength during preseason training in elite Australian footballers

Background Hamstring strain injuries (HSIs) are the most common injury type in Australian football and the rate of recurrence has been consistently high for a number of years. Long lasting neuromuscular inhibition has been noted in previously injured athletes but it is not known if this influences athletes adaptive response to training. Purpose To determine if elite Australian footballers with a prior unilateral HSI (previously injured group) display lesser improvements in eccentric hamstring strength during pre-season training compared to athletes without a history of HSI (control group). Study design Prospective cohort study. Methods Ninety-nine elite Australian footballers participated (17 with a history of unilateral HSI in the previous 12 month period). Eccentric hamstring strength was assessed at the start and end of pre-season training using an instrumented Nordic hamstring device. Change in eccentric strength across preseason was determine in absolute terms and normalised to start of preseason strength. Start of preseason strength was used as a covariate to control for differences in starting strength. Results The left and right limbs in the control group showed no difference in absolute or relative change (left limb absolute change, 60.7±72.9N; relative change, 1.28±0.34; right limb absolute change, 48.6±83.8N; relative change, 1.24±0.43) . Similarly, the injured and uninjured limbs from the previously injured group showed no difference for either absolute or relative measures of change (injured limb absolute change, 13.1±57.7N; relative change, 1.07±0.18; uninjured limb absolute change, 14.7±54.0N; relative change, 1.07±0.22N). The previously injured group displayed a significantly lesser increase in eccentric hamstring strength across the preseason (absolute change, 13.9±55.0; relative change, 1.07±0.20) compared to the control group (absolute change, 54.6±78.5; relative change, 1.26±0.39) for both absolute and relative measures (p < 0.001), even after controlling for differences in start of pre-season eccentric hamstring strength, which had a significant effect on strength improvement. Conclusion Elite Australian footballers with a unilateral HSI history displayed lesser improvements in eccentric hamstring strength across preseason training. The smaller improvements were not restricted to the previously injured limb as the contralateral limb also displayed similarly small improvements in eccentric strength. Whether this is the cause of or the result of injury remains to be seen, but it has the potential to contribute to the risk of hamstring strain re-injury.

The use of new generation geosynthetics in water treatment

The importance of clean drinking water in any community is absolutely vital if we as the consumers are to sustain a life of health and wellbeing. Suspended particles in surface waters not only provide the means to transport micro-organisms which can cause serious infections and diseases, they can also affect the performance capacity of a water treatment plant. In such situations pre-treatment ahead of the main plant is recommended. Previous research carried out using non-woven synthetic as a pre-filter materials for protecting slow sand filters from high turbidity showed that filter run times can be extended by several times and filters can be regenerated by simply removing and washing of the fabric ( Mbwette and Graham, 1987 and Mbwette, 1991). Geosynthetic materials have been extensively used for soil retention and dewatering in geotechnical applications and little research exists for the application of turbidity reduction in water treatment. With the development of new materials in geosynthetics today, it was hypothesized that the turbidity removal efficiency can be improved further by selecting appropriate materials. Two different geosynthetic materials (75 micron) tested at a filtration rate of 0.7 m/h yielded 30-45% reduction in turbidity with relatively minor head loss. It was found that the non-woven geotextile Propex 1701 retained the highest performance in both filtration efficiency and head loss across the varying turbidity ranges in comparison to other geotextiles tested. With 5 layers of the Propex 1701 an average percent reduction of approximately 67% was achieved with a head loss average of 4mm over the two and half hour testing period. Using the data collected for the Propex 1701 a mathematical model was developed for predicting the expected percent reduction given the ability to control the cost and as a result the number of layers to be used in a given filtration scenario.

Indigenous students transitioning to school : responses to pre-foundational mathematics

Australian Indigenous students' mathematics performance continues to be below that of non-Indigenous students. This occurs from the early years of school, due largely to knowledge and social differences on entry to formal schooling. This paper reports on a mathematics research project conducted in one Aboriginal community school in New South Wales, Australia. The project aimed to identify and explain the ways that young Australian Indigenous students (age 2-4 years) learn number language and processes, specifically attribute language, sorting, 1-1 correspondence and, counting. The project adopted a mixed methods approach. That is, the methodology was decolonising (Smith 1999) in that it collaborated with and gave benefit back to the Indigenous community and school being researched. It was qualitative and interpretative (Burns 2000) and incorporated an action-research teaching-experiment approach where and teachers collaborated with the researchers to try new teaching methods. This paper draws on data pertaining to students' response to diagnostic interview questions, the pre- and post-test results of the interview and photographic evidence as observations during mathematics learning time. Participants referred to in this paper include one female principal (N = 1), and the transition class of students' pre- (N = 6) and post-test (N = 3) results of the pre-foundational processes (also referred to as attributes). The results were encouraging with improvements in colour (34%), patterns (33%); capacity (38%). As a result of this project, our epistemology regarding the importance of finding out about students' pre-foundational knowledge and understandings and providing a culturally appropriate learning environment with resources has been built upon.

Durability performance of carbon fibre-reinforced polymer strengthened circular hollow steel members under cold weather

The use of circular hollow steel members has attracted a great deal of attention during past few years because of having excellent structural properties, aesthetic appearance, corrosion and fire protection capability. However, no one can deny the structural deficiency of such structures due to reduction of strength when they are exposed to severe environmental conditions such as marine environment, cold and hot weather. Hence strengthening and retrofitting of structural steel members is now very imperative. This paper presents the findings of a research program that was conducted to study the bond durability of carbon fibre-reinforced polymer (CFRP) strengthened steel tubular members under cold weather and tested under four-point bending. Six number of CFRP-strengthened specimens and one unstrengthened specimen were considered in this program. The three specimens having sand blasted surface to be strengthened was pre-treated with MBrace primer and other three were remained untreated and then cured under ambient temperature at least four weeks and cold weather (3 C) for three and six months period of time. Quasi-static tests were then performed on beams to failure under four-point bending. The structural response of each specimen was predicted in terms of failure load, mid-span deflection, composite beam behaviour and failure mode. The research outcomes show that the cold weather immersion had an adverse effect on durability of CFRP-strengthened steel structures. Moreover, the epoxy based adhesion promoter was found to enhance the bond durability in plastic range. The analytical models presented in this study were found to be in good agreement in terms of predicting ultimate load and deflection. Finally, design factors are proposed to address the short-terms durability performance under cold weather.