Photodissociation of TEMPO-modified peptides: New approaches to radical-directed dissociation of biomolecules

Radical-directed dissociation of gas phase ions is emerging as a powerful and complementary alternative to traditional tandem mass spectrometric techniques for biomolecular structural analysis. Previous studies have identified that coupling of 2-[(2,2,6,6-tetramethylpiperidin-1-oxyl)methyl] benzoic acid (TEMPO-Bz) to the N-terminus of a peptide introduces a labile oxygen-carbon bond that can be selectively activated upon collisional activation to produce a radical ion. Here we demonstrate that structurally-defined peptide radical ions can also be generated upon UV laser photodissociation of the same TEMPO-Bz derivatives in a linear ion-trap mass spectrometer. When subjected to further mass spectrometric analyses, the radical ions formed by a single laser pulse undergo identical dissociations as those formed by collisional activation of the same precursor ion, and can thus be used to derive molecular structure. Mapping the initial radical formation process as a function of photon energy by photodissociation action spectroscopy reveals that photoproduct formation is selective but occurs only in modest yield across the wavelength range (300-220 nm), with the photoproduct yield maximised between 235 and 225 nm. Based on the analysis of a set of model compounds, structural modifications to the TEMPO-Bz derivative are suggested to optimise radical photoproduct yield. Future development of such probes offers the advantage of increased sensitivity and selectivity for radical-directed dissociation. © 2014 the Owner Societies.

Publish

Our single-word issue title “publish” no doubt conjures up all sorts of anxieties in most writers and academics. It first rings through the head as a command to produce, to make, to compose—and underlines its necessity. Publish implies an invocation of engagement with its sister noun “public.” It also suggests an interchange and exchange between an audience of readers and the produced texts, leading to something that has been called a “sphere” in its grandiose claims and a “community” in its slightly more modest conceit. To publish is to produce a different form of conversation, one that is abstracted from the oral into the written and then presumably back out into both written and spoken, thereby producing new circuits of interchange and exchange....

Psychological predictors of injury among elite athletes

Objectives: To establish injury rates among a population of elite athletes, to provide normative data for psychological variables hypothesised to be predictive of sport injuries, and to establish relations between measures of mood, perceived life stress, and injury characteristics as a precursor to introducing a psychological intervention to ameliorate the injury problem. Methods: As part of annual screening procedures, athletes at the Queensland Academy of Sport report medical and psychological status. Data from 845 screenings (433 female and 412 male athletes) were reviewed. Population specific tables of normative data were established for the Brunel mood scale and the perceived stress scale. Results: About 67% of athletes were injured each year, and about 18% were injured at the time of screening. Fifty percent of variance in stress scores could be predicted from mood scores, especially for vigour, depression, and tension. Mood and stress scores collectively had significant utility in predicting injury characteristics. Injury status (current, healed, no injury) was correctly classified with 39% accuracy, and back pain with 48% accuracy. Among a subset of 233 uninjured athletes (116 female and 117 male), five mood dimensions (anger, confusion, fatigue, tension, depression) were significantly related to orthopaedic incidents over the preceding 12 months, with each mood dimension explaining 6–7% of the variance. No sex differences in these relations were found. Conclusions: The findings support suggestions that psychological measures have utility in predicting athletic injury, although the relatively modest explained variance highlights the need to also include underlying physiological indicators of allostatic load, such as stress hormones, in predictive models.

The 10 Australian ecosystems most vulnerable to tipping points

We identify the 10 major terrestrial and marine ecosystems in Australia most vulnerable to tipping points, in which modest environmental changes can cause disproportionately large changes in ecosystem properties. To accomplish this we independently surveyed the coauthors of this paper to produce a list of candidate ecosystems, and then refined this list during a 2-day workshop. The list includes (1) elevationally restricted mountain ecosystems, (2) tropical savannas, (3) coastal floodplains and wetlands, (4) coral reefs, (5) drier rainforests, (6) wetlands and floodplains in the Murray-Darling Basin, (7) the Mediterranean ecosystems of southwestern Australia, (8) offshore islands, (9) temperate eucalypt forests, and (10) salt marshes and mangroves. Some of these ecosystems are vulnerable to widespread phase-changes that could fundamentally alter ecosystem properties such as habitat structure, species composition, fire regimes, or carbon storage. Others appear susceptible to major changes across only part of their geographic range, whereas yet others are susceptible to a large-scale decline of key biotic components, such as small mammals or stream-dwelling amphibians. For each ecosystem we consider the intrinsic features and external drivers that render it susceptible to tipping points, and identify subtypes of the ecosystem that we deem to be especially vulnerable. © 2011 Elsevier Ltd.

Calcitonin receptor plays a physiological role to protect against hypercalcemia in mice

It is well established that calcitonin is a potent inhibitor of bone resorption; however, a physiological role for calcitonin acting through its cognate receptor, the calcitonin receptor (CTR), has not been identified. Data from previous genetically modified animal models have recognized a possible role for calcitonin and the CTR in controlling bone formation; however, interpretation of these data are complicated, in part because of their mixed genetic background. Therefore, to elucidate the physiological role of the CTR in calcium and bone metabolism, we generated a viable global CTR knockout (KO) mouse model using the Cre/loxP system, in which the CTR is globally deleted by >94% but <100%. Global CTRKOs displayed normal serum ultrafiltrable calcium levels and a mild increase in bone formation in males, showing that the CTR plays a modest physiological role in the regulation of bone and calcium homeostasis in the basal state in mice. Furthermore, the peak in serum total calcium after calcitriol [1,25(OH)2D3]-induced hypercalcemia was substantially greater in global CTRKOs compared with controls. These data provide strong evidence for a biological role of the CTR in regulating calcium homeostasis in states of calcium stress.

Tailored interventions based on exhaled nitric oxide versus clinical symptoms for asthma in children and adults

Background The measurement of severity and control of asthma in both children and adults can be based on subjective or objective measures. It has been advocated that fractional exhaled nitric oxide (FeNO) can be used to monitor airway inflammation as it correlates with some markers of asthma. Interventions for asthma therapies have been traditionally based on symptoms and/or spirometry. Objectives To evaluate the efficacy of tailoring asthma interventions based on exhaled nitric oxide in comparison to clinical symptoms (with or without spirometry/peak flow) for asthma related outcomes in children and adults. Search methods We searched the Cochrane Airways Group Specialised Register of Trials, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE and reference lists of articles. The last search was completed in February 2009. Selection criteria All randomised controlled comparisons of adjustment of asthma therapy based on exhaled nitric oxide compared to traditional methods (primarily clinical symptoms and spirometry/peak flow). Data collection and analysis Results of searches were reviewed against pre-determined criteria for inclusion. Relevant studies were independently selected in duplicate. Two authors independently assessed trial quality and extracted data. Authors were contacted for further information with response from one. Main results Two studies have been added for this update, which now includes six (2 adults and 4 children/adolescent) studies; these studies differed in a variety of ways including definition of asthma exacerbations, FeNO cut off levels, the way in which FeNO was used to adjust therapy and duration of study. Of 1053 participants randomised, 1010 completed the trials. In the meta-analysis, there was no significant difference between groups for the primary outcome of asthma exacerbations or for other outcomes (clinical symptoms, FeNO level and spirometry). In post-hoc analysis, a significant reduction in mean final daily dose inhaled corticosteroid per adult was found in the group where treatment was based on FeNO in comparison to clinical symptoms, (mean difference -450 mcg; 95% CI -677 to -223 mcg budesonide equivalent/day). However, the total amount of inhaled corticosteroid used in one of the adult studies was 11% greater in the FeNO arm. In contrast, in the paediatric studies, there was a significant increase in inhaled corticosteroid dose in the FeNO strategy arm (mean difference of 140 mcg; 95% CI 29 to 251, mcg budesonide equivalent/day). Authors' conclusions Tailoring the dose of inhaled corticosteroids based on exhaled nitric oxide in comparison to clinical symptoms was carried out in different ways in the six studies and found only modest benefit at best and potentially higher doses of inhaled corticosteroids in children. The role of utilising exhaled nitric oxide to tailor the dose of inhaled corticosteroids cannot be routinely recommended for clinical practice at this stage and remains uncertain.

The impact of viral respiratory infection on the severity and recovery from an asthma exacerbation

Background Viral respiratory illness triggers asthma exacerbations, but the influence of respiratory illness on the acute severity and recovery of childhood asthma is unknown. Our objective was to evaluate the impact of a concurrent acute respiratory illness (based on a clinical definition and PCR detection of a panel of respiratory viruses, Mycoplasma pneumoniae and Chlamydia pneumoniae) on the severity and resolution of symptoms in children with a nonhospitalized exacerbation of asthma. Methods Subjects were children aged 2 to 15 years presenting to an emergency department for an acute asthma exacerbation and not hospitalized. Acute respiratory illness (ARI) was clinically defined. Nasopharyngeal aspirates (NPA) were examined for respiratory viruses, Chlamydia and Mycoplasma using PCR. The primary outcome was quality of life (QOL) on presentation, day 7 and day 14. Secondary outcomes were acute asthma severity score, asthma diary, and cough diary scores on days 5, 7,10, and 14. Results On multivariate regression, presence of ARI was statistically but not clinically significantly associated with QOL score on presentation (B = 0.36, P = 0.025). By day 7 and 14, there was no difference between groups. Asthma diary score was significantly higher in children with ARI (B = 0.41, P = 0.039) on day 5 but not on presentation or subsequent days. Respiratory viruses were detected in 54% of the 78 NPAs obtained. There was no difference in the any of the asthma outcomes of children grouped by positive or negative NPA. Conclusions The presence of a viral respiratory illness has a modest influence on asthma severity, and does not influence recovery from a nonhospitalized asthma exacerbation.

Apple skin patterning is associated with differential expression of MYb10

Background Some apple (Malus × domestica Borkh.) varieties have attractive striping patterns, a quality attribute that is important for determining apple fruit market acceptance. Most apple cultivars (e.g. 'Royal Gala') produce fruit with a defined fruit pigment pattern, but in the case of 'Honeycrisp' apple, trees can produce fruits of two different kinds: striped and blushed. The causes of this phenomenon are unknown. Results Here we show that striped areas of 'Honeycrisp' and 'Royal Gala' are due to sectorial increases in anthocyanin concentration. Transcript levels of the major biosynthetic genes and MYB10, a transcription factor that upregulates apple anthocyanin production, correlated with increased anthocyanin concentration in stripes. However, nucleotide changes in the promoter and coding sequence of MYB10 do not correlate with skin pattern in 'Honeycrisp' and other cultivars differing in peel pigmentation patterns. A survey of methylation levels throughout the coding region of MYB10 and a 2.5 Kb region 5' of the ATG translation start site indicated that an area 900 bp long, starting 1400 bp upstream of the translation start site, is highly methylated. Cytosine methylation was present in all three contexts, with higher methylation levels observed for CHH and CHG (where H is A, C or T) than for CG. Comparisons of methylation levels of the MYB10 promoter in 'Honeycrisp' red and green stripes indicated that they correlate with peel phenotypes, with an enrichment of methylation observed in green stripes. Conclusions Differences in anthocyanin levels between red and green stripes can be explained by differential transcript accumulation of MYB10. Different levels of MYB10 transcript in red versus green stripes are inversely associated with methylation levels in the promoter region. Although observed methylation differences are modest, trends are consistent across years and differences are statistically significant. Methylation may be associated with the presence of a TRIM retrotransposon within the promoter region, but the presence of the TRIM element alone cannot explain the phenotypic variability observed in 'Honeycrisp'. We suggest that methylation in the MYB10 promoter is more variable in 'Honeycrisp' than in 'Royal Gala', leading to more variable color patterns in the peel of this cultivar.

Age and gender differences in objectively measured physical activity in youth

Purpose The purpose of this study was to evaluate age and gender differences in objectively measured physical activity (PA) in a population-based sample of students in grades 1–12. Methods Participants (185 male, 190 female) wore a CSA 7164 accelerometer for 7 consecutive days. To examine age-related trends, students were grouped as follows: grades 1–3 (N = 90), grades 4–6 (N = 91), grades 7–9 (N = 96), and grades 10–12 (N = 92). Bouts of PA and minutes spent in moderate-to-vigorous PA (MVPA) and vigorous PA (VPA) were examined. Results Daily MVPA and VPA exhibited a significant inverse relationship with grade level, with the largest differences occurring between grades 1–3 and 4–6. Boys were more active than girls; however, for overall PA, the magnitudes of the gender differences were modest. Participation in continuous 20-min bouts of PA was low to nonexistent. Conclusion Our results support the notion that PA declines rapidly during childhood and adolescence and that accelerometers are feasible alternatives to self-report methods in moderately sized population-level surveillance studies.

Measuring and influencing physical activity with Smartphone technology : a systematic review

Background Rapid developments in technology have encouraged the use of smartphones in physical activity research, although little is known regarding their effectiveness as measurement and intervention tools. Objective This study systematically reviewed evidence on smartphones and their viability for measuring and influencing physical activity. Data Sources Research articles were identified in September 2013 by literature searches in Web of Knowledge, PubMed, PsycINFO, EBSCO, and ScienceDirect. Study Selection The search was restricted using the terms (physical activity OR exercise OR fitness) AND (smartphone* OR mobile phone* OR cell phone*) AND (measurement OR intervention). Reviewed articles were required to be published in international academic peer-reviewed journals, or in full text from international scientific conferences, and focused on measuring physical activity through smartphone processing data and influencing people to be more active through smartphone applications. Study Appraisal and Synthesis Methods Two reviewers independently performed the selection of articles and examined titles and abstracts to exclude those out of scope. Data on study characteristics, technologies used to objectively measure physical activity, strategies applied to influence activity; and the main study findings were extracted and reported. Results A total of 26 articles (with the first published in 2007) met inclusion criteria. All studies were conducted in highly economically advantaged countries; 12 articles focused on special populations (e.g. obese patients). Studies measured physical activity using native mobile features, and/or an external device linked to an application. Measurement accuracy ranged from 52 to 100 % (n = 10 studies). A total of 17 articles implemented and evaluated an intervention. Smartphone strategies to influence physical activity tended to be ad hoc, rather than theory-based approaches; physical activity profiles, goal setting, real-time feedback, social support networking, and online expert consultation were identified as the most useful strategies to encourage physical activity change. Only five studies assessed physical activity intervention effects; all used step counts as the outcome measure. Four studies (three pre–post and one comparative) reported physical activity increases (12–42 participants, 800–1,104 steps/day, 2 weeks–6 months), and one case-control study reported physical activity maintenance (n = 200 participants; >10,000 steps/day) over 3 months. Limitations Smartphone use is a relatively new field of study in physical activity research, and consequently the evidence base is emerging. Conclusions Few studies identified in this review considered the validity of phone-based assessment of physical activity. Those that did report on measurement properties found average-to-excellent levels of accuracy for different behaviors. The range of novel and engaging intervention strategies used by smartphones, and user perceptions on their usefulness and viability, highlights the potential such technology has for physical activity promotion. However, intervention effects reported in the extant literature are modest at best, and future studies need to utilize randomized controlled trial research designs, larger sample sizes, and longer study periods to better explore the physical activity measurement and intervention capabilities of smartphones.

Oncogene-induced basement membrane invasiveness in human mammary epithelial cells

Expression of the intermediate filament protein vimentin, and loss of the cellular adhesion protein uvomorulin (E-cadherin) have been associated with increased invasiveness of established human breast cancer cell lines in vitro and in vivo. In the current study, we have further examined these relationships in oncogenically transformed human mammary epithelial cells. A normal human mammary epithelial strain, termed 184, was previously immortalized with benzo[a]pyrene, and two distinct sublines were derived (A1N4 and 184B5). These sublines were infected with retroviral vectors containing a single or two oncogenes of the nuclear, cytoplasmic, and plasma membrane-associated type (v-rasH, v-rasKi, v -mos, SV40T and c -myc). All infectants have been previously shown to exhibit some aspects of phenotypic transformation. In the current study, cellular invasiveness was determined in vitro using Matrigel, a reconstituted basement membrane extract. Lineage-specific differences were observed with respect to low constitutive invasiveness and invasive changes after infection with ras, despite similar ras-induced transformation of each line. Major effects on cellular invasiveness were observed after infection of the cells with two different oncogenes (v-rasH + SV40T and v -rasH + v -mos). In contrast, the effects of single oncogenes were only modest or negligible. All oncogenic infectants demonstrated increased attachment to laminin, but altered secretion of the 72 kDa and 92 kDa gelatinases was not associated with any aspect of malignant progression. Each of the two highly invasive double oncogene transformants were vimentinpositive and uvomorulin-negative, a phenotype indicative of the epithelial-mesenchymal transition (EMT) previously associated with invasiveness of established human breast cancer cell lines. Weakly invasive untransformed mammary epithelial cells in this study were positive for both vimentin and uvomorulin, suggesting that uvomorulin may over-ride the otherwise vimentin-associated invasiveness.

New approaches to the synthesis of strapped porphyrin containing bipyridinium [2]rotaxanes

Inspired by the interesting photo- and electrochemical properties observed in bipyridinium and porphyrin containing interlocked catenanes, herein we describe new approaches towards the synthesis of related rotaxanes. Previous efforts in this domain had been hampered by the limited range of chemical reactions that are compatible with these motifs, however the use of a “click” methodology, together with a better understanding of the size of these strapped porphyrin macrocycles, resulted in the synthesis of a bipyridinium porphyrin [2]rotaxane in modest yields. X-ray crystallography of the zinc metalloporphyrin macrocycle used in this study revealed that in the solid state, these strapped porphyrins adopt a 1-dimensional coordination polymer, in which an oxygen atom in the strap of one macrocycle is coordinated to the zinc metal center in an adjacent porphyrin ring

A series-connected photovoltaic distributed generator capable of enhancing power quality

Design of a series-connected photovoltaic generator (SPVG) capable of enhancing power quality is investigated. Analysis of the SPVG operations under disturbance conditions shows explicitly how achievable network voltage quality is affected by the SPVG injected power and its apparent power rating, and that voltage quality can be significantly improved even with a modest level of energy storage capacity incorporated in the SPVG. A control system for the SPVG is also proposed. Both simulation and laboratory tests confirm the efficacy of the distributed generator system.

The Pareto regret frontier

Performance guarantees for online learning algorithms typically take the form of regret bounds, which express that the cumulative loss overhead compared to the best expert in hindsight is small. In the common case of large but structured expert sets we typically wish to keep the regret especially small compared to simple experts, at the cost of modest additional overhead compared to more complex others. We study which such regret trade-offs can be achieved, and how. We analyse regret w.r.t. each individual expert as a multi-objective criterion in the simple but fundamental case of absolute loss. We characterise the achievable and Pareto optimal trade-offs, and the corresponding optimal strategies for each sample size both exactly for each finite horizon and asymptotically.

The Hollywood effect : a comparative study of costume in Australia and the Proposition

This paper compares costuming practices in Baz Luhrmann’s Australia (2008) and John Hillcoat’s The Proposition (2005) and argues that high production values, such as in the blockbuster Australia, are not neutral mechanisms of production, but powerful prescriptive elements which do not result in a successful representation of cultural specificity. Australia is a typical blockbuster, it employs a large number of extras, it features compelling landscape shots, has been shot across four different locations and sets, and, importantly, is an international production with the 20th Century Fox. The film’s costumes were designed by Catherine Martin, who received an Oscar nomination in 2009. While global exposure of fashion in film and through celebrities’ endorsements has consolidated a historical synergy between the fashion industry and Hollywood, the Australian film and fashion industries have had a very limited exchange. Baz Luhrmann’s film is Australia’s first instance of promo-costuming and use of tie-in labels (Ferragamo, R.M.Williams, Prada, Paspaley). Catherine Martin thoroughly researched 1930s women’s wear, indigenous and stockmen’s clothing, and set up to make all costumes with a large team of costumiers and seamstresses, striving for authenticity. The Proposition won its costume designer Margot Wilson an AFI in 2005 for best costume, but compared to Australia the story, location and costumes are far harsher. Filmed around Winton in far west Queensland, the director John Hillcoat and Director of Photography Benoit Delhomme were insistent about realism, and emphasising the harshness of the Australian landscape. The realism of the costumes was derived from the fabrics and manufacturing, as well as the way they were shot, with the actors often wearing two or three layers of heavy wool during days of shooting in 50 degree heat, and the details of making and breaking down. The implication is that both films are culturally specific as they both deal with an Australian story. However, Australia is clearly produced according to a Hollywood blockbuster model, and closely matches Hollywood’s narrative and aesthetic characteristics, while The Proposition is a more modest film that eschews these conventions of beauty and glossed history. Despite its western genre-orientation, The Proposition is more successful than Australia when it comes to costuming, because its costumes are not only functional to the narrative, but, in Roland Barthes’ words, they also fulfil a prestation. This prestation highlights the social and cultural conflicts on which colonial Australia was founded, instead of gilding, and gliding, over them.