Analysis of focal adhesion turnover: A quantitative live-cell imaging example

Recent advances in optical and fluorescent protein technology have rapidly raised expectations in cell biology, allowing quantitative insights into dynamic intracellular processes like never before. However, quantitative live-cell imaging comes with many challenges including how best to translate dynamic microscopy data into numerical outputs that can be used to make meaningful comparisons rather than relying on representative data sets. Here, we use analysis of focal adhesion turnover dynamics as a straightforward specific example on how to image, measure, and analyze intracellular protein dynamics, but we believe this outlines a thought process and can provide guidance on how to understand dynamic microcopy data of other intracellular structures.

Criminal histories of crash and non-crash involved Queensland speeding offenders: Evidence supporting the idea that we drive as we live

Evidence increasingly suggests that our behaviour on the road mirrors our behaviour across other aspects of our life. The idea that we drive as we live, described by Tillman and Hobbs more than 65 years ago when examining off-road behaviours of taxi drivers (1949), is the focus of the current paper. As part of a larger study examining the impact of penalty changes on a large cohort of Queensland speeding offenders, criminal (lifetime) and crash history (10 year period) data for a sub-sample of 1000 offenders were obtained. Based on the ‘drive as we live’ maxim, it was hypothesised that crash-involved speeding offenders would be more likely to have a criminal history than non-crash involved offenders. Overall, only 30% of speeding offenders had a criminal history. However, crash-involved offenders were significantly more likely to have a criminal history (49.4%) than non-crash involved offenders (28.6%), supporting the hypothesis. Furthermore, those deemed ‘most at fault’ in a crash were the group most likely to have at least one criminal offence (52.2%). When compared to the non-crash involved offenders, those deemed ‘not most at fault’ in a crash were also more likely to have had at least one criminal offence (46.5%). Therefore, when compared to non-crash involved speeding offenders, those offenders involved in a crash were more likely to have been convicted of at least one criminal offence, irrespective of whether they were deemed ‘most at fault’ in that crash. Implications for traffic offender management and policing are discussed.

Genome sequencing of cells that live inside glass cages reveals their past history

One-quarter of the total primary production on earth is contributed by diatoms1. These are photosynthetic, unicellular algae with ornamented silica shells found in all aquatic and moist environments. They form the base of energy-efficient food webs that support all aquatic life forms. More than 250 genera of living diatoms, with as many as 100,000 species are known2. Fossil diatoms are known as early as the Cretaceous, 144–65 m.y. ago3. In India, deposits of diatoms occur in Rajasthan and are known as ‘multani mitti’. Multani mitti or Indian Fuller’s earth or diatomaceous earth as it is called in the West, is applied as a paste on the surface of the skin for 15–20 min and then washed-off. This leaves the skin feeling smooth, soft, moist and rejuvenated. Diatomaceous earth is now being used in the formulation of soaps, cleansing products, face powders and skincare preparations. Diatomaceous earth is a mineral material consisting mainly of siliceous fragments of various species of fossilized remains of diatoms.

sopB of Salmonella enterica serovar Typhimurium is a potential DNA vaccine candidate in conjugation with live attenuated bacteria

The immune response against Salmonella is multi-faceted involving both the innate and the adaptive immune system. The characterization of specific Salmonella antigens inducing immune response could critically contribute to the development of epitope based vaccines for Salmonella. We have tried to identify a protective T cell epitope(s) of Salmonella, as cell mediated immunity conferred by CD8+ T cells is the most crucial subset conferring protective immunity against Salmonella. It being a proven fact that secreted proteins are better in inducing cell mediated immunity than cell surface and cytosolic antigens, we have analyzed all the genbank annotated Salmonella pathogenicity island 1 and 2 secreted proteins of Salmonella enterica serovar Typhimurium (S. typhimurium) and S. enterica serovar Typhi (S. typhi). They were subjected to BIMAS and SYFPEITHI analysis to map MHC-I and MHC-II binding epitopes. The huge profile of possible T cell epitopes obtained from the two classes of secreted proteins were tabulated and using a scoring system that considers the binding affinity and promiscuity of binding to more than one allele, SopB and SifB were chosen for experimental confirmation in murine immunization model. The entire SopB and SifB genes were cloned into DNA vaccine vectors and were administered along with live attenuated Salmonella and it was found that SopB vaccination reduced the bacterial burden of organs by about 5-fold on day 4 and day 8 after challenge with virulent Salmonella and proved to be a more efficient vaccination strategy than live attenuated bacteria alone.

“We live in in a house of blo- in a block of flats.” – Self-repair in EFL Spoken Language

Tämä tutkielma on osa Helsingin yliopiston rahoittamaa HY-talk -tutkimusprojektia, jonka tavoite on vankentaa puheviestinnän, erityisesti vieraiden kielten suullisen taidon opetusta ja arviointia yleissivistävässä koulutuksessa ja korkeakouluasteella. Tämän tutkielman tavoite on selvittää millaisia korjauksia englantia vieraana kielenä puhuvat ihmiset tekevät puheeseensa ja tutkia itsekorjauksen ja sujuvuuden välistä suhdetta. Korjausjäsennystä ja itsekorjausta on aiemmin tutkittu sekä keskustelunanalyysin että psykolingvistiikan aloilla, ja vaikka tämä tutkielma onkin lähempänä aiempaa keskustelunanalyyttistä kuin psykolingvististä tutkimusta, siinä hyödynnetään molempia suuntauksia. Itsekorjausta on yleisesti pidetty merkkinä erityisesti ei-natiivien kielenpuhujien sujuvuuden puutteesta. Tämän tutkielman tarkoitus on selvittää, kuinka läheisesti itsekorjaus todella liittyy sujuvuuteen tai sen puutteeseen. Tutkielman materiaali koostuu HY-talk -projektia varten kerätyistä puhenäytteistä ja niiden pohjalta tehdyistä taitotasoarvioinneista. Puhenäytteet kerättiin vuonna 2007 projektia varten järjestettyjen puhekielen testaustilanteiden yhteydessä kolmessa eteläsuomalaisessa koulussa. Koska projektin tavoitteena on tutkia ja parantaa kielten suullisen taidon arviointia, projektissa mukana olleet kieliammattilaiset arvioivat puhujien taitotasot projektia varten (Eurooppalaisen Viitekehyksen taitotasokuvainten pohjalta) koottujen arviointiasteikoiden perusteella, ja nämä arvioinnit tallennettiin osaksi projektin materiaalia. Tutkielmassa analysoidaan itsekorjauksia aiemman psykolingvistisen tutkimuksen pohjalta kootun korjaustyyppiluokituksen sekä tätä tutkielmaa varten luodun korjausten oikeellisuutta vertailevan luokituksen avulla. Lisäksi siinä vertaillaan kahden korkeamman ja kahden matalamman taitotasoarvioinnin saaneen puhujan itsekorjauksia. Tulokset osoittavat, että ei-natiivien puheessa esiintyy monenlaisia eri korjaustyyppejä, ja että yleisimpiä korjauksia ovat alkuperäisen lausuman toistot. Yleisiä ovat myös korjaukset, joissa puhuja korjaa virheen tai keskeyttää puheensa ja aloittaa kokonaan uuden lausuman. Lisäksi tuloksista käy ilmi, ettei suurin osa korjauksista todennäköisesti johdu puhujien sujuvuuden puutteesta. Yleisimmät korjaustyypit voivat johtua suurimmaksi osaksi yksilön puhetyylistä, siitä, että puhuja hakee jotain tiettyä sanaa tai ilmausta mielessään tai siitä, että puhuja korjaa puheessaan huomaamansa kieliopillisen, sanastollisen tai äänteellisen virheen. Vertailu korkeammalle ja matalammalle taitotasolle arvioitujen puhujien välillä osoittaa selkeimmin, ettei suurin osa itsekorjauksista ole yhteydessä puhujan sujuvuuteen. Vertailusta käy ilmi, ettei pelkkä itsekorjausten määrä kerro kuinka sujuvasti puhuja käyttää kieltä, sillä toinen korkeammalle taitotasolle arvioiduista puhujista korjaa puhettaan lähes yhtä monesti kuin matalammalle tasolle arvioidut puhujat. Lisäksi korjausten oikeellisuutta vertailevan luokituksen tulokset viittaavat siihen, etteivät niin korkeammalle kuin matalammallekaan tasolle arvioidut puhujat useimmiten korjaa puhettaan siksi, etteivät pystyisi ilmaisemaan viestiään oikein ja ymmärrettävästi.

Intake, growth and carcass characteristics of lambs consuming low digestible hay and cereal grain

The influence of barley and oat grain supplements on hay dry matter intake (DMI), carcass components gain and meat quality in lambs fed a low quality basal diet was examined. Thirty five crossbred wether lambs (9 months of age) were divided into four groups. After adaptation to a basal diet of 85% oat hay and 15% lucerne hay for one week, an initial group of 11 was slaughtered. The weights of carcass components and digesta-free empty body weight (EBW) of this group was used to estimate the weight of carcass components of the other three experimental groups at the start of the experiment. The remaining three groups were randomly assigned to pens and fed ad libitum the basal diet alone (basal), basal with 300 g air dry barley grain (barley), basal with 300 g air dry oat grain (oat). Supplements were fed twice weekly (i.e., 900 g on Tuesday and 1200 g on Friday). After 13 weeks of feeding, animals were slaughtered and, at 24 h post-mortem meat quality and subcutaneous fat colour were measured. Samples of longissimus muscle were collected for determination of sarcomere length and meat tenderness. Hay DMI was reduced (P<0.01) by both barley and oat supplements. Lambs fed barley or oat had a higher and moderate digestibility of DM, and a higher intake of CP (P<0.05) and ME (P<0.01) than basal lambs. Final live weight of barley and oat lambs was higher (P<0.05) than basal, but this was not reflected in EBW or hot carcass weight. Lambs fed barley or oat had increases in protein (P<0.01) and water (P<0.001) in the carcass, but fat gain was not changed (P>0.05). There were no differences in eye muscle area or fat depth (total muscle and adipose tissue depth at 12th rib, 110 mm from midline; GR) among groups. The increased levels of protein and water components in the carcass of barley and oat fed lambs, associated with improved muscle production, were small and did not alter (P>0.05) any of the carcass/meat quality attributes compared to lambs fed a low quality forage diet. Feeding barley or oat grain at 0.9–1% of live weight daily to lambs consuming poor quality hay may not substantially improve carcass quality, but may be useful in maintaining body condition of lambs through the dry season for slaughter out of season

Vaccination with an Insulin-like Growth Factor Binding Protein-1 (IGFBP-1)-bBased Vaccine Improves Growth in Feed-Restricted Brahman Steers

Dry-season weight loss in grazing cattle in northern Australia has been attenuated using a number of strategies (Hunter and Vercoe, 1987, Sillence et al. 1993, Gazzola and Hunter, 1999). Furthermore, the potential to improve efficiency of feed utilisation (and thus, dry-season performance) in ruminants through conventional modulation of the insulin-like growth factor (IGF) axis (Oddy and Owens, 1997, Hill et al., 1999) and through immunomodulation of the IGF axis (Hill et al., 1998a,b) has been demonstrated. The present study investigated the use of a vaccine directed against IGFBP-1 in Brahman steers which underwent a period of nutritional restriction followed by a return to wet-season grazing.

The Expanding Live Cattle Trade in Northern Australian

The north Australian beef industry is complex and dynamic. It is strategically positioned to access new and existing export markets. To prosper in a global economy, it will require strong processing and live cattle sectors, continued rationalisation of infrastructure, uptake of appropriate technology, and the synergy obtained when industry sectors unite and cooperate to maintain market advantage. Strategies to address food safety, animal welfare, the environment and other consumer concerns must be delivered. Strategic alliances with quality assurance systems will develop. These alliances will be based on economies of scale and on vertical cooperation, rather than vertical integration. Industry sectors will need to increase their contribution to Research, Development and Extension. These contributions need to be global in outlook. Industry sectors should also be aware that change (positive or negative) in one sector will impact on other sectors. Feedback along the food chain is essential to maximise productivity and market share.

Influence of nitrogen based supplements on live weight, fertility and mortality of heifers grazing dry season native pasture

Supplements containing urea or biuret were fed in the dry season to yearling and two year old pregnant heifers grazing native spear grass pastures in north Queensland. Liveweight change and survival during the dry season and fertility in the following year were measured. In the first experiment during a relatively favourable dry season, supplementation significantly (P<0.01) reduced liveweight loss in yearling heifers (5 vs. 32 kg). In the following year during a drought, supplement significantly (P<.01) reduced liveweight loss in yearling heifers (32 vs. 41 kg) and significantly (P <0.01) reduced mortalities (23.5% vs. 5.2%) in pregnant and lactating heifers. The supplement had no significant effect on subsequent fertility in either experiment. 14th Biennial Conference.

Selection and characterisation of two attenuated vaccine lines of Eimeria tenella in Australia

Objective To attenuate two strains of Eimeria tenella by selecting for precocious development and evaluate the strains in characterisation trials and by field evaluation, to choose one precocious line for incorporation into an Australian live coccidiosis vaccine for poultry. Design Two strains from non-commercial flocks were passaged through chickens while selecting for precocious development. Each strain was characterised for drug sensitivity, pathogenicity, protection against homologous and heterologous challenge, and oocyst output in replicated experiments in which the experimental unit was a cage of three birds. Oocyst output and/or body weight gain data collected over a 10 to 12 day period following final inoculation were measured. Feed conversion ratios were also calculated where possible. Results Fifteen passages resulted in prepatent periods reduced by 24 h for the Redlands strain (from 144 h to 120 h)and 23 h for the Darryl strain (from 139 h to 116 h). Characterisation trials demonstrated that each precocious line was significantly less pathogenic than its parent strain and each effectively induced immunity that protected chickens against challenge with both the parent strain and other virulent field strains. Both lines had oocyst outputs that, although significantly reduced relative to the parent strains, remained sufficiently high for commercial vaccine production, and both showed susceptibility to coccidiostats. Conclusion Two attenuated lines have been produced that exhibit the appropriate characteristics for use in an Australian live coccidiosis vaccine.

The reliability of supply of feed grains in the northern region

Reliability of supply of feed grain has become a high priority issue for industry in the northern region. Expansion by major intensive livestock and industrial users of grain, combined with high inter-annual variability in seasonal conditions, has generated concern in the industry about reliability of supply. This paper reports on a modelling study undertaken to analyse the reliability of supply of feed grain in the northern region. Feed grain demand was calculated for major industries (cattle feedlots, pigs, poultry, dairy) based on their current size and rate of grain usage. Current demand was estimated to be 2.8Mt. With the development of new industrial users (ethanol) and by projecting the current growth rate of the various intensive livestock industries, it was estimated that demand would grow to 3.6Mt in three years time. Feed grain supply was estimated using shire scale yield prediction models for wheat and sorghum that had been calibrated against recent ABS production data. Other crops that contribute to a lesser extent to the total feed grain pool (barley, maize) were included by considering their production relative to the major winter and summer grains, with estimates based on available production records. This modelling approach allowed simulation of a 101-year time series of yield that showed the extent of the impact of inter-annual climate variability on yield levels. Production estimates were developed from this yield time series by including planted crop area. Area planted data were obtained from ABS and ABARE records. Total production amounts were adjusted to allow for any export and end uses that were not feed grain (flour, malt etc). The median feed grain supply for an average area planted was about 3.1Mt, but this varied greatly from year to year depending on seasonal conditions and area planted. These estimates indicated that supply would not meet current demand in about 30% of years if a median area crop were planted. Two thirds of the years with a supply shortfall were El Nino years. This proportion of years was halved (i.e. 15%) if the area planted increased to that associated with the best 10% of years. Should demand grow as projected in this study, there would be few years where it could be met if a median crop area was planted. With area planted similar to the best 10% of years, there would still be a shortfall in nearly 50% of all years (and 80% of El Nino years). The implications of these results on supply/demand and risk management and investment in research and development are briefly discussed.

Assessing for genetic and environmental effects on ruminant feed quality in barley ( Hordeum vulgare )

Grain samples from a combined intermediate and advanced stage barley breeding trial series, grown at two sites in two consecutive years were assessed for detailed grain quality and ruminant feed quality. The results indicated that there were significant genetic and environmental effects for “feed” traits as measured using grain hardness, acid detergent fibre (ADF), starch and in-sacco dry matter digestibility (ISDMD) assays. In addition, there was strong genotypic discrimination for the regressed feed performance traits, namely Net Energy (NE) and Average Daily Gain (ADG). There was considerable variation in genetic correlations for all traits based on variance from the cultivars used, sites or laboratory processing effects. There was a high level of heritability ranging from 89% to 88% for retention, 60% to 80% for protein and 56% to 68% for ADF. However, there were only low to moderate levels of heritability for the feed traits, with starch 30–39%, ISDMD 55–63%, ADF 56–68%, particle size 47–73%, 31–48% NE and ADG 44–51%. These results suggest that there were real differences in the feed performance of barleys and that selection for cattle feed quality is potentially a viable option for breeding programs.

How effective are broad-scale nutrient mass balances for determining the sustainability of lot-feed manure application?

Nutrient mass balances have been used to assess a variety of land resource scenarios, at various scales. They are widely used as a simple basis for policy, planning, and regulatory decisions but it is not clear how accurately they reflect reality. This study provides a critique of broad-scale nutrient mass balances, with particular application to the fertiliser use of beef lot-feeding manure in Queensland. Mass balances completed at the district and farm scale were found to misrepresent actual manure management behaviour and potentially the risk of nutrient contamination of water resources. The difficulties of handling stockpile manure and concerns about soil compaction mean that manure is spread thickly over a few paddocks at a time and not evenly across a whole farm. Consequently, higher nutrient loads were applied to a single paddock less frequently than annually. This resulted in years with excess nitrogen, phosphorus, and potassium remaining in the soil profile. This conclusion was supported by evidence of significant nutrient movement in several of the soil profiles studied. Spreading manure is profitable, but maximum returns can be associated with increased risk of nutrient leaching relative to conventional inorganic fertiliser practices. Bio-economic simulations found this increased risk where manure was applied to supply crop nitrogen requirements (the practice of the case study farms, 200-5000 head lot-feeders). Thus, the use of broad-scale mass balances can be misleading because paddock management is spatially heterogeneous and this leads to increased local potential for nutrient loss. In response to the effect of spatial heterogeneity policy makers who intend to use mass balance techniques to estimate potential for nutrient contamination should apply these techniques conservatively.

Influence of food supplementation on the fitness of two biological control agents: a predatory nabid bug and a bollworm pupal parasitoid

Many arthropod predators and parasitoids exhibit either stage-specific or lifetime omnivory, in that they include extra-floral nectar, floral nectar, honeydew or pollen in their immature and/or adult diet. Access to these plant-derived foods can enhance pest suppression by increasing both the individual fitness and local density of natural enemies. Commercial products such as Amino-Feed®, Envirofeast®, and Pred-Feed® can be applied to crops to act as artificial-plant-derived foods. In laboratory and glasshouse experiments we examined the influence of carbohydrate and protein rich Amino-Feed UV® or Amino-Feed, respectively, on the fitness of a predatory nabid bug Nabis kinbergii Reuter (Hemiptera: Nabidae) and bollworm pupal parasitoid Ichneumon promissorius (Erichson) (Hymenoptera: Ichneumonidae). Under the chosen conditions, the provision of either wet or dry residues of Amino-Feed UV had no discernable effect on immediate or longer-term survival and immature development times of N. kinbergii. In contrast, the provision of honey, Amino-Feed plus extrafloral nectar, and extrafloral nectar alone had a marked effect on the longevity of I. promissorius, indicating that they were limited by at least carbohydrates as an energy source, but probably not protein. Compared with a water only diet, the provision of Amino-Feed plus extrafloral nectar increased the longevity of males and females of I. promissorius by 3.0- and 2.4-fold, respectively. Not only did female parasitoids live longer when provided food, but the total number of eggs laid and timing of deposition was affected by diet under the chosen conditions. Notably, females in the water and honey treatments deposited greater numbers of eggs earlier in the trial, but this trend was unable to be sustained over their lifetime. Egg numbers in these treatments subsequently fell below the levels achieved by females in the Amino-Feed plus extrafloral nectar and cotton extrafloral nectar only treatments. Furthermore, there were times when the inclusion of the Amino-Feed was beneficial compared with cotton extrafloral nectar only. Artificial food supplements and plant-derived foods are worthy of further investigation because they have potential to improve the ecosystem service of biological pest control in targeted agroecosystems by providing natural enemies with an alternative source of nutrition, particularly during periods of prey/host scarcity.

The physiological and metabolic impacts on sheep and cattle of feed and water deprivation before and during transport

Sheep and cattle are frequently subjected to feed and water deprivation (FWD) for about 12 h before, and then during, transport to reduce digesta load in the gastrointestinal tract. This FWD is marked by weight loss as urine and faeces mainly in the first 24 h but continuing at a reduced rate subsequently. The weight of rumen contents falls although water loss is to some extent masked by saliva inflow. FWD is associated with some stress, particularly when transportation is added. This is indicated by increased levels of plasma cortisol that may be partly responsible for an observed increase in the output of water and N in urine and faeces. Loss of body water induces dehydration that may induce feelings of thirst by effects on the hypothalamus structures through the renin-angiotensin-aldosterone system. There are suggestions that elevated cortisol levels depress angiotensin activity and prevent sensations of thirst in dehydrated animals, but further research in this area is needed. Dehydration coupled with the discharge of Na in urine challenges the maintenance of homeostasis. In FWD, Na excretion in urine is reduced and, with the reduction in digesta load, Na is gradually returned from the digestive tract to the extracellular fluid space. Control of enteropathogenic bacteria by normal rumen microbes is weakened by FWD and resulting infections may threaten animal health and meat safety. Recovery time is required after transport to restore full feed intake and to ensure that adequate glycogen is present in muscle pre-slaughter to maintain meat quality.