A protocol for the rapid isolation of full geminivirus genomes from dried plant tissue

A high-throughput method of isolating and cloning geminivirus genomes from dried plant material, by combining an Extract-n-Amp™-based DNA isolation technique with rolling circle amplification (RCA) of viral DNA, is presented. Using this method an attempt was made to isolate and clone full geminivirus genomes/genome components from 102 plant samples, including dried leaves stored at room temperature for between 6 months and 10 years, with an average hands-on-time to RCA-ready DNA of 15 min per 20 samples. While storage of dried leaves for up to 6 months did not appreciably decrease cloning success rates relative to those achieved with fresh samples, efficiency of the method decreased with increasing storage time. However, it was still possible to clone virus genomes from 47% of 10-year-old samples. To illustrate the utility of this simple method for high-throughput geminivirus diversity studies, six Maize streak virus genomes, an Abutilon mosaic virus DNA-B component and the DNA-A component of a previously unidentified New Word begomovirus species were fully sequenced. Genomic clones of the 69 other viruses were verified as such by end sequencing. This method should be extremely useful for the study of any circular DNA plant viruses with genome component lengths smaller than the maximum size amplifiable by RCA. © 2008 Elsevier B.V. All rights reserved.

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.

Muscle, skin and core temperature after −110°C cold air and 8°C water treatment

The aim of this investigation was to elucidate the reductions in muscle, skin and core temperature following exposure to −110°C whole body cryotherapy (WBC), and compare these to 8°C cold water immersion (CWI). Twenty active male subjects were randomly assigned to a 4-min exposure of WBC or CWI. A minimum of 7 days later subjects were exposed to the other treatment. Muscle temperature in the right vastus lateralis (n = 10); thigh skin (average, maximum and minimum) and rectal temperature (n = 10) were recorded before and 60 min after treatment. The greatest reduction (P<0.05) in muscle (mean ± SD; 1 cm: WBC, 1.6±1.2°C; CWI, 2.0±1.0°C; 2 cm: WBC, 1.2±0.7°C; CWI, 1.7±0.9°C; 3 cm: WBC, 1.6±0.6°C; CWI, 1.7±0.5°C) and rectal temperature (WBC, 0.3±0.2°C; CWI, 0.4±0.2°C) were observed 60 min after treatment. The largest reductions in average (WBC, 12.1±1.0°C; CWI, 8.4±0.7°C), minimum (WBC, 13.2±1.4°C; CWI, 8.7±0.7°C) and maximum (WBC, 8.8±2.0°C; CWI, 7.2±1.9°C) skin temperature occurred immediately after both CWI and WBC (P<0.05). Skin temperature was significantly lower (P<0.05) immediately after WBC compared to CWI. The present study demonstrates that a single WBC exposure decreases muscle and core temperature to a similar level of those experienced after CWI. Although both treatments significantly reduced skin temperature, WBC elicited a greater decrease compared to CWI. These data may provide information to clinicians and researchers attempting to optimise WBC and CWI protocols in a clinical or sporting setting.

Effects of average speed enforcement on speed compliance and crashes : a review of the literature

Average speed enforcement is a relatively new approach gaining popularity throughout Europe and Australia. This paper reviews the evidence regarding the impact of this approach on vehicle speeds, crashes rates and a number of additional road safety and public health outcomes. The economic and practical viability of the approach as a road safety countermeasure is also explored. A literature review, with an international scope, of both published and grey literature was conducted. There is a growing body of evidence to suggest a number of road safety benefits associated with average speed enforcement, including high rates of compliance with speed limits, reductions in average and 85th percentile speeds and reduced speed variability between vehicles. Moreover, the approach has been demonstrated to be particularly effective in reducing excessive speeding behaviour. Reductions in crash rates have also been reported in association with average speed enforcement, particularly in relation to fatal and serious injury crashes. In addition, the approach has been shown to improve traffic flow, reduce vehicle emissions and has also been associated with high levels of public acceptance. Average speed enforcement offers a greater network-wide approach to managing speeds that reduces the impact of time and distance halo effects associated with other automated speed enforcement approaches. Although comparatively expensive it represents a highly reliable approach to speed enforcement that produces considerable returns on investment through reduced social and economic costs associated with crashes.

Average travel time estimations for urban routes that consider exit turning movements

This paper presents a methodology for real-time estimation of exit movement-specific average travel time on urban routes by integrating real-time cumulative plots, probe vehicles, and historic cumulative plots. Two approaches, component based and extreme based, are discussed for route travel time estimation. The methodology is tested with simulation and is validated with real data from Lucerne, Switzerland, that demonstrate its potential for accurate estimation. Both approaches provide similar results. The component-based approach is more reliable, with a greater chance of obtaining a probe vehicle in each interval, although additional data from each component is required. The extreme-based approach is simple and requires only data from upstream and downstream of the route, but the chances of obtaining a probe that traverses the entire route might be low. The performance of the methodology is also compared with a probe-only method. The proposed methodology requires only a few probes for accurate estimation; the probe-only method requires significantly more probes.

Preliminary evaluation of a primary care intervention for cry-fuss behaviours in the first 3-4 months of life (‘The Possums Approach’) : effects on cry-fuss behaviours and maternal mood

Problem crying in the first few months of life is both common and complex, arising out of multiple interacting and co-evolving factors. Parents whose babies cry and fuss a lot receive conflicting advice as they seek help from multiple health providers and emergency departments, and may be admitted into tertiary residential services. Conflicting advice is costly, and arises out of discipline-specific interpretations of evidence. An integrated, interdisciplinary primary care intervention (‘The Possums Approach’) for cry-fuss problems in the first months of life was developed from available peer-reviewed evidence. This study reports on preliminary evaluation of delivery of the intervention. A total of 20 mothers who had crying babies under 16 weeks of age (average age 6.15 weeks) completed questionnaires, including the Crying Patterns Questionnaire and the Edinburgh Postnatal Depression Scale, before and 3-4 weeks after their first consultation with trained primary care practitioners. Preliminary evaluation is promising. The Crying Patterns Questionnaire showed a significant decrease in crying and fussing duration, by 1 h in the evening (P = 0.001) and 30 min at night (P = 0.009). The median total amount of crying and fussing in a 24-h period was reduced from 6.12 to 3 h. The Edinburgh Postnatal Depression Scale showed a significant improvement in depressive symptoms, with the median score decreasing from 11 to 6 (P = 0.005). These findings are corroborated by an analysis of results for the subset of 16 participants whose babies were under 12 weeks of age (average age 4.71 weeks). These preliminary results demonstrate significantly decreased infant crying in the evening and during the night and improved maternal mood, validating an innovative interdisciplinary clinical intervention for cry-fuss problems in the first few months of life. This intervention, delivered by trained health professionals, has the potential to mitigate the costly problem of health professionals giving discipline-specific and conflicting advice post-birth.

Effects of whole body cryotherapy and cold water immersion on knee skin temperature

This study sought to a) compare and contrast the effect of 2 commonly used cryotherapy treatments, 4 min of − 110 °C whole body cryotherapy and 8 °C cold water immersion, on knee skin temperature and b) establish whether either protocol was capable of achieving a skin temperature ( < 13 °C) believed to be required for analgesic purposes. After ethics committee approval and written informed consent was obtained, 10 healthy males (26.5 ± 4.9 yr, 183.5 ± 6.0 cm, 90.7 ± 19.9 kg, 26.8 ± 5.0 kg/m 2 , 23.0 ± 9.3 % body fat; mean ± SD) participated in this randomised controlled crossover study. Skin temperature around the patellar region was assessed in both knees via non-contact, infrared thermal imaging and recorded pre-, immediately post-treatment and every 10 min thereafter for 60 min. Compared to baseline, average, minimum and maximum skin temperatures were significantly reduced (p < 0.001) immediately post-treatment and at 10, 20, 30, 40, 50 and 60 min after both cooling modalities. Average and minimum skin temperatures were lower (p < 0.05) immediately after whole body cryotherapy (19.0 ± 0.9 ° C) compared to cold water immersion (20.5 ± 0.6 ° C). However, from 10 to 60 min post, the average, minimum and maximum skin temperatures were lower (p < 0.05) following the cold water treatment. Finally, neither protocol achieved a skin temperature believed to be required to elicit an analgesic effect.

Count-min sketches for estimating password frequency within Hamming distance two

The count-min sketch is a useful data structure for recording and estimating the frequency of string occurrences, such as passwords, in sub-linear space with high accuracy. However, it cannot be used to draw conclusions on groups of strings that are similar, for example close in Hamming distance. This paper introduces a variant of the count-min sketch which allows for estimating counts within a specified Hamming distance of the queried string. This variant can be used to prevent users from choosing popular passwords, like the original sketch, but it also allows for a more efficient method of analysing password statistics.

The role of intensity of interval training on fat oxidation and eating behaviour in overweight/obese men

The increasing prevalence of obesity in society has been associated with a number of atherogenic risk factors such as insulin resistance. Aerobic training is often recommended as a strategy to induce weight loss, with a greater impact of high-intensity levels on cardiovascular function and insulin sensitivity, and a greater impact of moderate-intensity levels on fat oxidation. Anaerobic high-intensity (supramaximal) interval training has been advocated to improve cardiovascular function, insulin sensitivity and fat oxidation. However, obese individuals tend to have a lower tolerance of high-intensity exercise due to discomfort. Furthermore, some obese individuals may compensate for the increased energy expenditure by eating more and/or becoming less active. Recently, both moderate- and high-intensity aerobic interval training have been advocated as alternative approaches. However, it is still uncertain as to which approach is more effective in terms of increasing fat oxidation given the issues with levels of fitness and motivation, and compensatory behaviours. Accordingly, the objectives of this thesis were to compare the influence of moderate- and high-intensity interval training on fat oxidation and eating behaviour in overweight/obese men. Two exercise interventions were undertaken by 10-12 overweight/obese men to compare their responses to study variables, including fat oxidation and eating behaviour during moderate- and high-intensity interval training (MIIT and HIIT). The acute training intervention was a methodological study designed to examine the validity of using exercise intensity from the graded exercise test (GXT) - which measured the intensity that elicits maximal fat oxidation (FATmax) - to prescribe interval training during 30-min MIIT. The 30-min MIIT session involved 5-min repetitions of workloads 20% below and 20% above the FATmax. The acute intervention was extended to involve HIIT in a cross-over design to compare the influence of MIIT and HIIT on eating behaviour using subjective appetite sensation and food preference through the liking and wanting test. The HIIT consisted of 15-sec interval training at 85 %VO2peak interspersed by 15-sec unloaded recovery, with a total mechanical work equal to MIIT. The medium term training intervention was a cross-over 4-week (12 sessions) MIIT and HIIT exercise training with a 6-week detraining washout period. The MIIT sessions consisted of 5-min cycling stages at ±20% of mechanical work at 45 %VO2peak, and the HIIT sessions consisted of repetitive 30-sec work at 90 %VO2peak and 30-sec interval rests, during identical exercise sessions of between 30 and 45 mins. Assessments included a constant-load test (45 %VO2peak for 45 mins) followed by 60-min recovery at baseline and the end of 4-week training, to determine fat oxidation rate. Participants’ responses to exercise were measured using blood lactate (BLa), heart rate (HR) and rating of perceived exertion (RPE) and were measured during the constant-load test and in the first intervention training session of every week during training. Eating behaviour responses were assessed by measuring subjective appetite sensations, liking and wanting and ad libitum energy intake. Results of the acute intervention showed that FATmax is a valid method to estimate VO2 and BLa, but is not valid to estimate HR and RPE in the MIIT session. While the average rate of fat oxidation during 30-min MIIT was comparable with the rate of fat oxidation at FATmax (0.16 ±0.09 and 0.14 ±0.08 g/min, respectively), fat oxidation was significantly higher at minute 25 of MIIT (P≤0.01). In addition, there was no significant difference between MIIT and HIIT in the rate of appetite sensations after exercise, but there was a tendency towards a lower rate of hunger after HIIT. Different intensities of interval exercise also did not affect explicit liking or implicit wanting. Results of the medium-term intervention indicated that current interval training levels did not affect body composition, fasting insulin and fasting glucose. Maximal aerobic capacity significantly increased (P≤0.01) (2.8 and 7.0% after MIIT and HIIT respectively) during GXT, and fat oxidation significantly increased (P≤0.01) (96 and 43% after MIIT and HIIT respectively) during the acute constant-load exercise test. RPE significantly decreased after HIIT greater than MIIT (P≤0.05), and the decrease in BLa was greater during the constant-load test after HIIT than MIIT, but this difference did not reach statistical significance (P=0.09). In addition, following constant-load exercise, exercise-induced hunger and desire to eat decreased after HIIT greater than MIIT but were not significant (p value for desire to eat was 0.07). Exercise-induced liking of high-fat sweet (HFSW) and high-fat non-sweet (HFNS) foods increased after MIIT and decreased after HIIT (p value for HFNS was 0.09). The intervention explained 12.4% of the change in fat intake (p = 0.07). This research is significant in that it confirmed two points in the acute study. While the rate of fat oxidation increased during MIIT, the average rate of fat oxidation during 30-min MIIT was comparable with the rate of fat oxidation at FATmax. In addition, manipulating the intensity of acute interval exercise did not affect appetite sensations and liking and wanting. In the medium-term intervention, constant-load exercise-induced fat oxidation significantly increased after interval training, independent of exercise intensity. In addition, desire to eat, explicit liking for HFNS and fat intake collectively confirmed that MIIT is accompanied by a greater compensation of eating behaviour than HIIT. Findings from this research will assist in developing exercise strategies to provide obese men with various training options. In addition, the finding that overweight/obese men expressed a lower RPE and decreased BLa after HIIT compared with MIIT is contrary to the view that obese individuals may not tolerate high-intensity interval training. Therefore, high-intensity interval training can be advocated among the obese adult male population. Future studies may extend this work by using a longer-term intervention.

Travel time prediction on signalised urban arterials by applying SARIMA modelling on Bluetooth data

Travel time prediction has long been the topic of transportation research. But most relevant prediction models in the literature are limited to motorways. Travel time prediction on arterial networks is challenging due to involving traffic signals and significant variability of individual vehicle travel time. The limited availability of traffic data from arterial networks makes travel time prediction even more challenging. Recently, there has been significant interest of exploiting Bluetooth data for travel time estimation. This research analysed the real travel time data collected by the Brisbane City Council using the Bluetooth technology on arterials. Databases, including experienced average daily travel time are created and classified for approximately 8 months. Thereafter, based on data characteristics, Seasonal Auto Regressive Integrated Moving Average (SARIMA) modelling is applied on the database for short-term travel time prediction. The SARMIA model not only takes the previous continuous lags into account, but also uses the values from the same time of previous days for travel time prediction. This is carried out by defining a seasonality coefficient which improves the accuracy of travel time prediction in linear models. The accuracy, robustness and transferability of the model are evaluated through comparing the real and predicted values on three sites within Brisbane network. The results contain the detailed validation for different prediction horizons (5 min to 90 minutes). The model performance is evaluated mainly on congested periods and compared to the naive technique of considering the historical average.

Age-dependence of the average and equivalent refractive indices of the crystalline lens

Lens average and equivalent refractive indices are required for purposes such as lens thickness estimation and optical modeling. We modeled the refractive index gradient as a power function of the normalized distance from lens center. Average index along the lens axis was estimated by integration. Equivalent index was estimated by raytracing through a model eye to establish ocular refraction, and then backward raytracing to determine the constant refractive index yielding the same refraction. Assuming center and edge indices remained constant with age, at 1.415 and 1.37 respectively, average axial refractive index increased (1.408 to 1.411) and equivalent index decreased (1.425 to 1.420) with age increase from 20 to 70 years. These values agree well with experimental estimates based on different techniques, although the latter show considerable scatter. The simple model of index gradient gives reasonable estimates of average and equivalent lens indices, although refinements in modeling and measurements are required.

Access to commercial destinations within the neighbourhood and walking among Australian older adults

BACKGROUND: Physical activity, particularly walking, is greatly beneficial to health; yet a sizeable proportion of older adults are insufficiently active. The importance of built environment attributes for walking is known, but few studies of older adults have examined neighbourhood destinations and none have investigated access to specific, objectively-measured commercial destinations and walking. METHODS: We undertook a secondary analysis of data from the Western Australian state government's health surveillance survey for those aged 65--84 years and living in the Perth metropolitan region from 2003--2009 (n = 2,918). Individual-level road network service areas were generated at 400 m and 800 m distances, and the presence or absence of six commercial destination types within the neighbourhood service areas identified (food retail, general retail, medical care services, financial services, general services, and social infrastructure). Adjusted logistic regression models examined access to and mix of commercial destination types within neighbourhoods for associations with self-reported walking behaviour. RESULTS: On average, the sample was aged 72.9 years (SD = 5.4), and was predominantly female (55.9%) and married (62.0%). Overall, 66.2% reported some weekly walking and 30.8% reported sufficient walking (>=150 min/week). Older adults with access to general services within 400 m (OR = 1.33, 95% CI = 1.07-1.66) and 800 m (OR = 1.20, 95% CI = 1.02-1.42), and social infrastructure within 800 m (OR = 1.19, 95% CI = 1.01-1.40) were more likely to engage in some weekly walking. Access to medical care services within 400 m (OR = 0.77, 95% CI = 0.63-0.93) and 800 m (OR = 0.83, 95% CI = 0.70-0.99) reduced the odds of sufficient walking. Access to food retail, general retail, financial services, and the mix of commercial destination types within the neighbourhood were all unrelated to walking. CONCLUSIONS: The types of neighbourhood commercial destinations that encourage older adults to walk appear to differ slightly from those reported for adult samples. Destinations that facilitate more social interaction, for example eating at a restaurant or church involvement, or provide opportunities for some incidental social contact, for example visiting the pharmacy or hairdresser, were the strongest predictors for walking among seniors in this study. This underscores the importance of planning neighbourhoods with proximate access to social infrastructure, and highlights the need to create residential environments that support activity across the life course.

In vivo and in vitro models demonstrate a role for caveolin-1 in the pathogenesis of ischaemic acute renal failure

Caveolae and their proteins, the caveolins, transport macromolecules; compartmentalize signalling molecules; and are involved in various repair processes. There is little information regarding their role in the pathogenesis of significant renal syndromes such as acute renal failure (ARF). In this study, an in vivo rat model of 30 min bilateral renal ischaemia followed by reperfusion times from 4 h to 1 week was used to map the temporal and spatial association between caveolin-1 and tubular epithelial damage (desquamation, apoptosis, necrosis). An in vitro model of ischaemic ARF was also studied, where cultured renal tubular epithelial cells or arterial endothelial cells were subjected to injury initiators modelled on ischaemia-reperfusion (hypoxia, serum deprivation, free radical damage or hypoxia-hyperoxia). Expression of caveolin proteins was investigated using immunohistochemistry, immunoelectron microscopy, and immunoblots of whole cell, membrane or cytosol protein extracts. In vivo, healthy kidney had abundant caveolin-1 in vascular endothelial cells and also some expression in membrane surfaces of distal tubular epithelium. In the kidneys of ARF animals, punctate cytoplasmic localization of caveolin-1 was identified, with high intensity expression in injured proximal tubules that were losing basement membrane adhesion or were apoptotic, 24 h to 4 days after ischaemia-reperfusion. Western immunoblots indicated a marked increase in caveolin-1 expression in the cortex where some proximal tubular injury was located. In vitro, the main treatment-induced change in both cell types was translocation of caveolin-1 from the original plasma membrane site into membrane-associated sites in the cytoplasm. Overall, expression levels did not alter for whole cell extracts and the protein remained membrane-bound, as indicated by cell fractionation analyses. Caveolin-1 was also found to localize intensely within apoptotic cells. The results are indicative of a role for caveolin-1 in ARF-induced renal injury. Whether it functions for cell repair or death remains to be elucidated.

Investigation on mechanical behaviour of AM60 magnesium alloys

Purpose: In this work, tension, impact, bend and fatigue tests were conducted in an AM60 magnesium alloy. The effects of environmental temperature and loading rates on impact and tension behavior of the alloy were also investigated. Design/methodology/approach: The tests were conducted using an Instron universal testing machine. The loading speed was changed from 1 mm/min to 300 mm/min to gain a better understanding of the effect of strain rate. To understand the failure behavior of this alloy at different environmental temperatures, Charpy impact test was conducted in a range of temperatures (-40~35°C). Plane strain fracture toughness (KIC) was evaluated using compact tension (CT) specimen. To gain a better understanding of the failure mechanisms, all fracture surfaces were observed using scanning electron microscopy (SEM). In addition, fatigue behavior of this alloy was estimated using tension test under tension-tension condition at 30 Hz. The stress amplitude was selected in the range of 20~50 MPa to obtain the S-N curve. Findings: The tensile test indicated that the mechanical properties were not sensitive to the strain rates applied (3.3x10-4~0.1) and the plastic deformation was dominated by twining mediated slip. The impact energy is not sensitive to the environmental temperature. The plane strain fracture toughness and fatigue limit were evaluated and the average values were 7.6 MPa.m1/2 and 25 MPa, respectively. Practical implications: Tested materials AM60 Mg alloy can be applied among others in automotive industry aerospace, communication and computer industry. Originality/value: Many investigations have been conducted to develop new Mg alloys with improved stiffness and ductility. On the other hand, relatively less attention has been paid to the failure mechanisms of Mg alloys, such as brittle fracture and fatigue, subjected to different environmental or loading conditions. In this work, tension, impact, bend and fatigue tests were conducted in an AM60 magnesium alloy.

Intervening to reduce workplace sitting time : how and when do changes to sitting time occur?

Objective To investigate how and when changes in workplace sitting time occurred following a workplace intervention to inform evaluation of intervention success. Method The 4-week Stand Up Comcare study (June–September 2011) aimed to reduce workplace sitting time via regularly interrupting and replacing sitting time throughout the day. Activity monitor (activPAL3) workplace data from control (n=22) and intervention participants (n=21) were analysed. Differences in the number and usual duration of sitting bouts were used to evaluate how change occurred. To examine when change occurred, intervention effects were compared by hour since starting work and hour of the workday. Change in workplace activity (sitting, standing, stepping) was examined to further inform alignment with intervention messages. Individual variability was examined in how and when the change occurred. Results Overall, behavioural changes aligned with intervention aims. All intervention participants reduced total workplace sitting time, though there was wide individual variability observed (range −29 to −262 min per 8 h workday). On average, intervention participants reduced number of sitting bouts (−4.6 bouts (95% CI −10.1 to 1.0), p=0.106) and usual sitting bout duration (−5.6 min (95% CI −9.8 to −1.4, p=0.011)) relative to controls. Sitting time reductions were observed across the workday, though intervention effects varied by hour of the day (p=0.015). The intervention group successfully adopted the Stand Up and Sit Less intervention messages across the day. Conclusion These analyses confirmed that this workplace intervention successfully modified sitting behaviour as intended (ie, fewer and shorter sitting bouts, with changes occurring throughout the day).