Substrate stiffness promotes vascular smooth muscle cell calcification by reducing the levels of nuclear actin monomers.

BACKGROUND: Vascular calcification (VC) is a prevalent independent risk factor for adverse cardiovascular events and is associated with diabetes, hypertension, chronic kidney disease, and atherosclerosis. However, the mechanisms regulating the osteogenic differentiation of vascular smooth muscle cells (VSMC) are not fully understood. METHODS: Using hydrogels of tuneable stiffness and lysyl oxidase-mediated stiffening of human saphenous vein ex vivo, we investigated the role of substrate stiffness in the regulation of VSMC calcification. RESULTS: We demonstrate that increased substrate stiffness enhances VSMC osteogenic differentiation and VSMC calcification. We show that the effects of substrate stiffness are mediated via a reduction in the level of actin monomer within the nucleus. We show that in cells interacting with soft substrate, elevated levels of nuclear actin monomer repress osteogenic differentiation and calcification by repressing YAP-mediated activation of both TEA Domain transcription factor (TEAD) and RUNX Family Transcription factor 2 (RUNX2). CONCLUSION: This work highlights for the first time the role of nuclear actin in mediating substrate stiffness-dependent VSMC calcification and the dual role of YAP-TEAD and YAP-RUNX2 transcriptional complexes.

Survey of imposex in Nucella lapillus as an indicator of tributyltin pollution in Northern Irish coastal waters, 2004 to 2017.

Tributyltin (TBT) was a commonly used biocide in anti-fouling paints for marine vessels, however, it had negative impacts on non-target species, including causing imposex in dogwhelks (Nucella lapillus). Since the 1980s, legislation has been introduced to curtail its usage, culminating in a ban by the International Maritime Organisation (IMO) in 2008. In 2004 a national imposex monitoring network was established in Northern Ireland to determine the level and extent of TBT pollution. The level of imposex in N. lapillus was assessed according to the degree of male sexual development in the female using three measures: percentage of imposex-affected females (%I), the relative penis size index (RPSI), and the vas deferens sequence index (VDSI). All sites showed improvement since the initial survey with a reduction in imposex and an improvement in ecological quality under the classification of the Convention for the Protection of the Marine Environment of the North-East Atlantic (OSPAR).

Biological and ecological evidences suggest Stipa krylovii (Pooideae), contributes to optimal growth performance and population distribution of the grasshopper Oedaleus asiaticus.

Oedaleus asiaticus Bey. Bienko is a significant grasshopper pest species occurring in north Asian grasslands. Outbreaks often result in significant loss in grasses and economic losses. Interestingly, we found this grasshopper was mainly restricted to Stipa-dominated grassland. We suspected this may be related to the dominant grasses species, Stipa krylovii Roshev, and hypothesized that S. krylovii contributes to optimal growth performance and population distribution of O. asiaticus. A 4 year investigation showed that O. asiaticus density was positively correlated to the above-ground biomass of S. krylovii and O. asiaticus growth performance variables (survival rate, size, growth rate) were significantly higher in Stipa-dominated grassland. A feeding trial also showed that O. asiaticus had a higher growth performance when feeding exclusively on S. krylovii. In addition, the choice, consumption and the efficiency of conversion of ingested food (ECI) by O. asiaticus was highest for S. krylovii compared with other plant species found in the Asian grasslands. These ecological and biological traits revealed why O. asiaticus is strongly associated with Stipa-dominated grasslands. We concluded that the existence of S. krylovii benefited the growth performance and explained the distribution of O. asiaticus. These results are useful for improved pest management strategies and developing guidelines for the monitoring of grasshopper population dynamics against the background of vegetation succession and changing plant communities in response to activities such as grazing, fire and climate change.

Quantitative Analysis of Plant Consumption and Preference by Oedaleus asiaticus (Acrididae: Oedipodinae) in Changed Plant Communities Consisting of Three Grass Species.

We used Oedaleus asiaticus Bey-Bienko (Acrididae: Oedipodinae) as a model species to examine how plant community composition influenced grasshopper population densities and feeding habits. Our results show that in the Xinli Gol Grassland ecosystem, both plant community composition and grasshopper density vary considerably over fairly small distances, and that O. asiaticus density was directly related to the proportional abundance of specific plant species. A 60-d field-cage study showed that O. asiaticus food preference and consumption differed when the ratios of three dominant grass species (Leymus chinensis (Trin.) Tzvel, Stipa krylovii Roshev, and Cleistogenes squarrosa (Trin.) Keng) differed. The consumption of both S. krylovii and C. squarrosa increased with a corresponding increase in their respective biomass availability. Although O. asiaticus preferred S. krylovii and ate only small amounts of L. chinensis, a specific amount of L. chinensis was always consumed. Feeding of these three plant species could be divided into three levels based on diet selectivity index (SI), with O. asiaticus showing a strong preference for S. krylovii (SI > 1), over that for C. squarrosa (0.5 < SI ≤ 1), and a low acceptance for L. chinensis (0 < SI ≤ 0.5). Plant consumption and preference by O. asiaticus was in decreasing order S. krylovii>C. squarrosa>L. chinensis. This study shows that small-scale changes in plant community composition can have large affects on grasshopper diets and populations. As such, this study is useful for developing guidelines for the monitoring of O. asiaticus impacts against the background of vegetation succession and changing plant communities in response to grazing or human activities.

Brain regions and molecular pathways responding to food reward type and value in honey bees.

The ability of honey bees to evaluate differences in food type and value is crucial for colony success, but these assessments are made by individuals who bring food to the hive, eating little, if any, of it themselves. We tested the hypothesis that responses to food type (pollen or nectar) and value involve different subsets of brain regions, and genes responsive to food. mRNA in situ hybridization of c-jun revealed that brain regions responsive to differences in food type were mostly different from regions responsive to differences in food value, except those dorsal and lateral to the mushroom body calyces, which responded to all three. Transcriptomic profiles of the mushroom bodies generated by RNA sequencing gave the following results: (1) responses to differences in food type or value included a subset of molecular pathways involved in the response to food reward; (2) genes responsive to food reward, food type and food value were enriched for (the Gene Ontology categories) mitochondrial and endoplasmic reticulum activity; (3) genes responsive to only food and food type were enriched for regulation of transcription and translation; and (4) genes responsive to only food and food value were enriched for regulation of neuronal signaling. These results reveal how activities necessary for colony survival are channeled through the reward system of individual honey bees.

Methyl bromide fumigation and delayed mortality: safe trade of live pests?

Live organisms intercepted from treated commodities during phytosanitary inspections usually arouse suspicions of treatment failure, sub-standard treatment application, or post-treatment infestation. The additional possibility that some treatments could kill slowly, meaning commodities might be inspected before pests have succumbed, is seldom considered for treatments other than irradiation. We used a novel biochemical viability assay to measure delays between methyl bromide fumigation and mortality of dipteran eggs, and evaluated the correspondence between egg viability and egg morphological features. Our experimental conditions simulated shipping of rock melons from Australia to New Zealand by sea and air. No eggs survived fumigation, but they took 3-20 days to die, whereas phytosanitary inspections of rock melons occur within 2-7 days. Delays were not influenced by methyl bromide concentration, but were significantly lengthened by cooler storage temperatures. Methyl bromide's preservative effects delayed degradation of egg morphology, so the biochemical assay detected mortality long before morphological signs of egg death appeared. The results show that commodities subjected to effective methyl bromide treatments are at risk of being inspected before all pests have either died, or started to exhibit morphological signs of death. This could cause commodities to be unnecessarily rejected by quarantine authorities. Better methods than inspection for live pests are needed to assist authorities to gain assurance that treated commodities have been effectively disinfested. These could be developed by exploiting biochemical responses of pests and commodities to treatments.

Voxel-based analysis of the immediate early gene, c-jun, in the honey bee brain after a sucrose stimulus.

Immediate early genes (IEGs) have served as useful markers of brain neuronal activity in mammals, and more recently in insects. The mammalian canonical IEG, c-jun, is part of regulatory pathways conserved in insects and has been shown to be responsive to alarm pheromone in honey bees. We tested whether c-jun was responsive in honey bees to another behaviourally relevant stimulus, sucrose, in order to further identify the brain regions involved in sucrose processing. To identify responsive regions, we developed a new method of voxel-based analysis of c-jun mRNA expression. We found that c-jun is expressed in somata throughout the brain. It was rapidly induced in response to sucrose stimuli, and it responded in somata near the antennal and mechanosensory motor centre, mushroom body calices and lateral protocerebrum, which are known to be involved in sucrose processing. c-jun also responded to sucrose in somata near the lateral suboesophageal ganglion, dorsal optic lobe, ventral optic lobe and dorsal posterior protocerebrum, which had not been previously identified by other methods. These results demonstrate the utility of voxel-based analysis of mRNA expression in the insect brain.

Identification and electrophysiological studies of (4S,5S)-5-hydroxy-4-methyl-3-heptanone and 4-methyl-3,5-heptanedione in male lucerne weevils.

An investigation to identify a sex or aggregation pheromone of Sitona discoideus Gyllenhål (Coleoptera: Curculionidae) is presented. Antenna flicking and attraction behaviors evoked by conspecifics of both sexes were recorded in arena bioassays, where attraction of females to males was observed. Air entrainment of both males and females was conducted in separate chambers. Gas chromatographic-mass spectrometric analysis of headspace volatiles revealed that two male-specific compounds, 4-methyl-3,5-heptanedione (major) and (4S,5S)-5-hydroxy-4-methyl-3-heptanone (minor), were emitted during the autumnal post-aestivatory flight period. The stereoisomers of the minor component were separated by enantioselective gas chromatography and their absolute configurations assigned by NMR (diastereomers) and the known preference of enantioselective transesterification reactions catalyzed by Candida antarctica lipase B. Electroantennogram and single sensillum recording studies indicate that 4-methyl-3,5-heptanedione as well as all individual stereoisomers of 5-hydroxy-4-methyl-3-heptanone are detected by the antennae of male and female S. discoideus. Further, single sensillum recordings suggest that both sexes of S. discoideus have specialized olfactory receptor neurons (ORNs) for detecting 4-methyl-3,5-heptanedione and different populations of stereoselective ORNs for detecting the stereoisomers of 5-hydroxy-4-methyl-3-heptanone. Some of these stereoselective ORNs appear to be sex-specific in S. discoideus.

Virtual reality in the rehabilitation of the arm after hemiplegic stroke: a randomized controlled pilot study.

OBJECTIVE: To assess the feasibility of a trial to investigate the effectiveness of virtual reality-mediated therapy compared to conventional physiotherapy in the motor rehabilitation of the arm following stroke, and to provide data for a power analysis to determine numbers for a future main trial. DESIGN: Pilot randomized controlled trial. SETTING: Clinical research facility. PARTICIPANTS: Eighteen people with a first stroke, 10 males and 8 females, 7 right and 2 left side most affected. Mean time since stroke 10.8 months. INTERVENTIONS: Participants were randomized to a virtual reality group or a conventional arm therapy group for nine sessions over three weeks. MAIN MEASURES: The upper limb Motricity Index and the Action Research Arm Test were completed at baseline, post intervention and six weeks follow-up. RESULTS: Outcome data were obtained from 95% of participants at the end of treatment and at follow-up: one participant withdrew. Compliance was high; only two people reported side-effects from virtual reality exposure. Both groups demonstrated small (7-8 points on upper limb Motricity Index and 4 points on the Action Research Arm Test), but non-significant, changes to their arm impairment and activity levels. CONCLUSION: A randomized controlled trial of virtual reality-mediated therapy comparable to conventional therapy would be feasible, with some suggested improvements in recruitment and outcome measures. Seventy-eight participants (39 per group) would be required for a main trial.

Virtual reality in the rehabilitation of the upper limb after stroke: the user's perspective.

Our group has developed a relatively low-cost virtual reality (VR) system for rehabilitation of the upper limb following stroke. Our system is immersive in that the participant views a representation of their arm and hand, reaching and retrieving objects in the virtual environment (VE), through a head-mounted display (HMD). This is thought to increase the participant's sense of presence in the VE and may lead to improved rehabilitation outcomes. However, use of immersion, particularly with our low-cost system, may increase the incidence of side effects reported. Therefore, the aim of this project was to assess the interaction of healthy users and those following stroke, in terms of their experience of presence in the VE and the rate of self-reported side effects. Differences in rates of perceived exertion, levels of enjoyment, and sense of control between both groups were also explored.

Total and nematode-specific IgE responses in intestinal lymph of genetically resistant and susceptible sheep during infection with Trichostrongylus colubriformis.

Total and antigen-specific IgE responses in afferent (AIL) and efferent (EIL) intestinal lymph of sheep with a nematode resistant (R) or susceptible (S) genotype during challenge infection with the intestinal nematode parasite Trichostrongylus colubriformis were examined. Within each sheep line, lambs with a nematode naive or nematode field-primed pre-challenge status were used. Total IgE level in AIL and EIL was dependent on nematode infection and was further influenced by genotype or the immune phenotype (nematode immune mean FEC+/-SDM=77+/-179 or non-immune mean FEC+/-SDM=4016+/-4318) of the animal. During T. colubriformis challenge immune animals had higher levels of total IgE in lymph than non-immune sheep, R line sheep had higher concentrations of total IgE than S line sheep, and field-primed animals had higher total IgE levels than nematode naive animals. Concentrations of total IgE were consistently higher in AIL than EIL or serum and were higher in lymph draining the proximal than the distal jejunum demonstrating that polyclonal IgE in AIL was largely derived from the intestinal mucosa of the anatomical compartment where the nematodes reside. The consistently higher concentration of total IgE in AIL was dependent on phenotype or genotype and in S genotype sheep also on the pre-challenge status. Concentrations of nematode specific IgE were significantly higher in EIL than AIL indicating a preference for the production of IgE reacting with excretory secretory products of the infective T. colubriformis larvae in the regional lymph node.

Unexpected Listronotus bonariensis (Coleoptera: Curculionidae) mortality in the presence of parasitoids.

Field enclosure and laboratory cage experiments designed to measure the impact of the parasitoid Microctonus hyperodae Loan on the Argentine stem weevil, Listronotus bonariensis (Kuschel) have shown that under high parasitoid pressure, there is significant weevil mortality without obvious parasitism. Parasitoid-exposed, but unparasitized portions of caged populations died at rates significantly higher than both the parasitized weevils from the same population and the unexposed controls. Unexpectedly, parasitized weevils had the lowest mortality rates (prior to larval parasitoid emergence). It was of ecological significance that this mortality effect was detected under field conditions using several years of empirically-collected population dynamics data. Under both field and laboratory conditions this mortality was found to occur within shorter time intervals than that required for full parasitoid larval development; thus it could not be attributed to the mortality that occurs after the emergence of prepupal parasitoids. The mechanism for this mortality remains undetermined, although it could possibly be linked to pseudoparasitism.

Piezoelectric sensor determination of arterial pulse wave velocity.

Arterial pulse wave velocity (APWV) is a measure of the elasticity (or stiffness) of peripheral arterial blood vessels. The pulse referred to here will be the pressure pulse as opposed to the flow pulse measured by ultrasound Doppler. The pressure pulse velocity varies over the range from about 12 m s(-1) to 15 m s(-1) in stiff peripheral arteries, whereas in normal arteries it has a velocity in the range of 7 to 9 m s(-1). The aim of this project was the development of a fast and easy to use system for the determination of peripheral arterial pulse wave velocity. The principle of the PWV measurement is based on simultaneous measurement of two pulse waves at two different positions, such as the radial artery at the wrist and the brachial artery just above the elbow. By determining the pulse transit time between these points and the distance measured between the two locations, pulse wave velocity may then be calculated. The pressure pulse detection is done by using two piezoelectric sensors which generate a measurable voltage at the output contacts if they are mechanically deformed. The deformation produced voltage is first amplified and filtered and then digitalized with a data acquisition card. The analysis of the data obtained from the sensors includes a filtering process, the calculation of the PWV with three different methods--foot-to-foot, cross-correlation and peak-to-peak-and the determination of the arterial pulse rate. Extensive measurements with human test subjects were carried out to optimize the techniques of data acquisition and analysis. For example, it was found that the best procedure was to hold the sensors in place using elastic straps alone. The data analysis was upgraded with an additional software module, which deletes, in effect, outriders or invalid measurements. With the optimized system, a series involving eight test subjects ranging in age from 22 to 32 years was completed (all normotensive). The arterial pulse wave velocities determined covered a range from 6 m s(-1) to 12 m s(-1), with an average standard deviation of less than 2.5 m s(-1) for individual results. These are slightly higher, but close to published APWV data. The results showed that reproducible results can be obtained with the existing PWV acquirement and analysis system.

Immune rejection of Trichostrongylus colubriformis in sheep; a possible role for intestinal mucus antibody against an L3-specific surface antigen.

Sheep that have been immunized by multiple truncated infections with the parasitic nematode Trichostrongylus colubriformis contain anti-larval activity in their intestinal mucus and high-speed mucus supernatants. This activity induces T. colubriformis L3 to clump in vitro and causes a significant reduction in larval establishment in naive sheep after infusion of larvae and mucus into the intestinal lumen via a duodenal cannula. In this report, we provide evidence that one factor contributing to the anti-larval activity of immune mucus is antibody against a 35-kDa L3-specific cuticular antigen. The anti-larval activity in mucus is > 100 kDa by membrane filtration, is heat labile and sensitive to either protease digestion or reduction with DTT. Immunoblotting showed that mucus and supernatants of ultracentrifuged mucus from immune sheep contained IgG1 and IgA antibodies that recognized predominantly a larval antigen with an estimated molecular weight of 35 kDa on SDS-PAGE. Antibodies eluted from the surface of washed larvae that had been incubated in immune mucus also reacted specifically with the 35 kDa antigen on blots of larval homogenate. Immunofluorescence and immunogold electron microscopy showed that the 35 kDa antigen is present on the epicuticle of L3 and is shed during the moult to L4. The antigen is not present in eggs, L1, L2, L4 or adult worms and is found only in extracts of sheaths and L3 before infection and up to 4 days after infection. We hypothesize that the binding of antibody to the larval surface prevents larvae from establishing at their preferred site, causing them to be eliminated from the intestine. Monoclonal antibody PAB-1 recognizes the 35 kDa T. colubriformis larval antigen and also cross-reacts with antigens of similar molecular weight on blots of L3 extracts of the parasitic nematodes Haemonchus contortus and Ostertagia circumcincta; and with a 22-kDa antigen on blots of L3 extracts from Cooperia curticei and Nematodirus spathiger. This indicates that an antigenically related surface antigen with immunizing potential is present on several nematode species and can be identified by mAb PAB-1. The 35 kDa T. colubriformis larval antigen and related molecules in other nematodes are potential novel targets for stimulating host-protective immunity against nematode infections.

An analysis of consultations with the crowd doctors at Glasgow Celtic football club, season 1999-2000.

OBJECTIVE: To analyse all clinical presentations to the crowd doctors at Scotland's largest football stadium over the course of one complete season. METHODS: A standard clinical record form was used to document all consultations with the crowd doctors including treatment and subsequent referrals. The relevance of alcohol consumption was assessed. RESULTS: A total of 127 casualties were seen at 26 matches, a mean of 4.88 per match. Twenty casualties were transferred to hospital, including one successfully defibrillated after a cardiac arrest. Alcohol excess was a major contributing factor in 26 cases. CONCLUSIONS: The workload of the crowd doctors was very variable and diverse. The social problem of excessive alcohol consumption contributed considerably to the workload. The provision of medical facilities at football grounds means that attendance there is now one of the least adverse circumstances in which to have a cardiac arrest. The study confirmed previous impressions that more casualties are seen at high profile matches.

Glycopyrrolate reduces nausea during spinal anaesthesia for caesarean section without affecting neonatal outcome.

We have tested the hypotheses that glycopyrrolate, administered immediately before induction of subarachnoid anaesthesia for elective Caesarean section, reduces the incidence and severity of nausea, with no adverse effects on neonatal Apgar scores, in a double-blind, randomized, controlled study. Fifty women received either glycopyrrolate 200 micrograms or saline (placebo) i.v. during fluid preload, before induction of spinal anaesthesia with 2.5 ml of 0.5% isobaric bupivacaine. Patients were questioned directly regarding nausea at 3-min intervals throughout operation and asked to report symptoms as they arose. The severity of nausea was assessed using a verbal scoring system and was treated with increments of i.v. ephedrine and fluids. Patients in the group pretreated with glycopyrrolate reported a reduction in the frequency (P = 0.02) and severity (P = 0.03) of nausea. Glycopyrrolate also reduced the severity of hypotension, as evidenced by reduced ephedrine requirements (P = 0.02). There were no differences in neonatal Apgar scores between groups.

Appropriateness of international heat stress standards for use in tropical agricultural environments.

Where a danger to health from heat stress is identified, standards allow decisions for implementing measures to reduce the heat stress to be made. These standards, specifically ISO 7243 (Wet Bulb Globe Temperature Index, WBGT) and ISO 7933 (Sweat Required, SWreq) were designed with European and American subjects, primarily for use in those countries. While the scope of the standards is international, little consideration has been made about how valid and usable they are in industrially developing countries. This investigation evaluated ISO 7933 and ISO 7243 in terms of validity and usability. A tropical agricultural task was simulated; 16 subjects plucking tea leaves for 2 h, (ta = tr = 37.18 degrees C; va = 0.16 m/s; rh = 70.17%). While ISO 7243 was valid (if slightly over protective) and usable, ISO 7933 was over protective and underestimated sweat and evaporation rates in its predictions. The discrepancies between predicted and observed results were attributed primarily to the calculations related to clothing in the standard. Furthermore, ISO 7933 was unusable without a computer; in regions where access to such technology may be limited, a simpler method of presentation is required.