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Extracellular vesicles (EVs) are small lipid-enclosed particles that can carry various types of cargo, including proteins, nucleic acids and metabolites. They are known to be released by all cell types and can be taken up by other cells, leading to the transfer of the cargo they carry. As such, they represent an important type of intercellular signalling and a natural mechanism for transferring macromolecules between cells. This ability to transfer cargo could be harnessed to deliver therapeutic molecules. Indeed, a growing body of work has described the attempt by the field to utilise EVs to deliver a range of therapeutics including RNAi, CRISPR/Cas9 and chemotherapeutics, to a specific target tissue. However, there are numerous barriers associated with the use of EVs as therapeutic vehicles, including the challenge of efficiently loading therapeutics into EVs, avoiding clearance of the EVs from circulation, targeting the correct tissue type and the inefficiency of internalisation and functional delivery of the cargo. Despite these difficulties, EVs represent a tremendous therapeutic opportunity, both for the delivery of exogenous cargo, as well as the therapeutic benefit of targeting aberrant EV signalling or treating patients with natural EVs, such as those released by mesenchymal stem cells. This review describes current knowledge on the therapeutic potential of EVs and the challenges faced by the field. Many of these challenges are due to a lack of complete understanding of EV function, but further research in this area should continue to yield new solutions that will lead to the use of EVs in the clinic.

Introduction Aberrant function of granulosa cells has been implicated in the pathophysiology of PCOS. Materials & methods Granulosa lutein (GL) cells were collected during oocyte retrieval for IVF/ICSI. RT-qPCR was used to compare gene expression between 12 control women, 12 with ovulatory PCO and 12 with anovulatory PCOS. To examine which genes are directly regulated by androgens, GL cells from an additional 12 control women were treated in-vitro with 10 nM dihydrotestosterone (DHT). Results GL cells from women with PCOS showed reduced expression of CYP11A1 3-fold (p = 0.005), HSD17B1 1.8-fold (p = 0.02) and increased expression of SULT1E1 7-fold (p = 0.0003). Similar results were seen in ovulatory women with PCO. GL cells treated with 10 nM DHT showed a 4-fold (p = 0.03) increase in expression of SULT1E1 and a 5-fold reduction in SRD5A1 (p = 0.03). Conclusions These findings support the notion that aberrant regulation of steroid metabolism or action play a part in ovarian dysfunction in PCOS.

To quantify head and neck cancer (HNC) mortality rates and identify racial and socioeconomic factors associated with 90-day mortality.The National Cancer Database (2004-2014) was queried for eligible HNC cases (n = 260,011) among adults treated with curative intent. Outcome of interest was any-cause 90-day mortality. Kaplan-Meier curves (Log-rank tests) estimated crude survival differences. A Cox proportional hazards model with further adjustments using the Šidák multiple comparison method adjusted for racial, socioeconomic and clinical factors.There were 9771 deaths (90-day mortality rate = 3.8%). There were crude differences in sex, race/ethnicity, comorbidity, distance, income, and insurance (Log-rank p-value 0.0001). In the final model, blacks (aHR = 1.10, 95% CI 1.00, 1.21) and males (aHR = 1.07; 95% CI 1.00, 1.15) had greater 90-day mortality hazard, as did those uninsured (aHR = 1.72; 95% CI 1.48, 1.99), covered by Medicaid (aHR = 1.72; 95% CI 1.53, 1.93) or Medicare (aHR = 1.40; 95% CI 1.27, 1.53). Residence in lower median income zip code was associated with greater 90-day mortality [(aHR$30,000 = 1.30; 95% CI 1.18, 1.44); (aHR $30,000-$34,999 = 1.24; 95% CI 1.13, 1.36); (aHR $35,000-$45,999 = 1.18; 95% CI 1.08, 1.27)]; and farther travel distance for treatment was associated with decreased 90-day mortality [(aHR 50-249.9 miles = 0.86; 95% CI 0.77, 0.97); (aHR 250 miles = 0.70; 95% CI 50, 0.99)].There are significant race and socioeconomic disparities among patients with HNC, and these disparities impact mortality within 90 days of treatment.

Weight suppression (WS), the difference in one's highest and current adult weight excluding pregnancy, is associated with eating disorder (ED) symptom onset and maintenance. There is increasing evidence that WS is related to ED symptoms in non-clinical samples, but there are no known studies examining the interaction of WS and current body mass index (BMI) in this group. History of overweight is common in those with EDs, and higher weight status often delays identification and treatment. This study examined the interaction of WS and current BMI on body dissatisfaction and eating pathology in undergraduate men and women. Undergraduates (N = 476) completed online surveys assessing current height and weight, weight history, and eating attitudes and behaviors. In women (n = 333), both BMI and WS were positively associated with body dissatisfaction, restraint, and global ED symptoms. In men (n = 143), there were no significant associations with BMI, but WS was related to greater body dissatisfaction, restraint, global ED symptoms, and loss-of-control eating. Moderation analyses revealed that the effect of current BMI on shape concern (p = .005), weight concern (p = .002), global ED symptoms (p = .01), and purging behaviors (p = .04) in women and restraint (p = .03) in men weakened at the highest levels of WS. Results suggest that WS is salient in a non-clinical sample and underscore the need to evaluate both weight history and the presence of disordered eating in individuals across the BMI spectrum.

Genetic adaptation to captivity influences the reproductive success of hatchery-reared steelhead (Oncorhynchus mykiss) spawning in the wild, but the mechanism by which adaptation occurs is not well understood. Because body size-at-release is positively correlated with post-release survival in hatchery-reared smolts, one hypothesis is that hatcheries select for physiological and/or behavioral traits that promote fast growth in captivity. If those juvenile growth traits are maladaptive for spawning or offspring survival in the wild, then that could explain why hatchery fish quickly evolve to have lower reproductive success than natural-origin fish in the wild environment. First-generation hatchery-reared steelhead from the Hood River, (Oregon) have lower reproductive success in the wild than do natural-origin fish, and substantial evidence suggests the fitness difference is genetically based and due to genetic adaptation to captivity. Here we ask whether the ‘selection on size-at-release’ hypothesis could explain the rapid genetic adaptation observed in this well-studied steelhead population. Using scale analysis, we back-calculated length at ocean entrance to test whether size-selective survival occurred in two cohorts (brood year [BY] 1997 and 2009). In BY 2009, we found evidence of weak size-selective survival (difference of 9 mm between pre-release average length and back-calculated length from surviving adults), but in BY 1997, strong (37 mm difference) size-selective survival was observed. Family identity explained 33% of the variance in fork length before release, and fork length was highly heritable in both BY cohorts. Thus, the requisite genetic variation for response to selection on size-at-release exists in this population. Our results support the hypothesis that size-selective survival does occur after release, and that selection for traits promoting fast growth in the hatchery could be a mechanism by which rapid adaptation to captivity has occurred in the Hood River winter steelhead hatchery program.

The organization of cnidarian nerve nets is traditionally described as diffuse with randomly arranged neurites that show minimal reproducibility between animals. However, most observations of nerve nets are conducted using cross-reactive antibodies that broadly label neurons, which potentially masks stereotyped patterns produced by individual neuronal subtypes. Additionally, many cnidarians species have overt structures such as a nerve ring, suggesting higher levels of organization and stereotypy exist, but mechanisms that generated that stereotypy are unknown. We previously demonstrated that NvLWamide-like is expressed in a small subset of the Nematostella nerve net and speculated that observing a few neurons within the developing nerve net would provide a better indication of potential stereotypy. Here we document NvLWamide-like expression more systematically. NvLWamide-like is initially expressed in the typical neurogenic salt and pepper pattern within the ectoderm at the gastrula stage, and expression expands to include endodermal salt and pepper expression at the planula larval stage. Expression persists in both ectoderm and endoderm in adults. We characterized our NvLWamide-like::mCherry transgenic reporter line to visualize neural architecture and found that NvLWamide-like is expressed in six neural subtypes identifiable by neural morphology and location. Upon completing development the numbers of neurons in each neural subtype are minimally variable between animals and the projection patterns of each subtype are consistent. Furthermore, between the juvenile polyp and adult stages the number of neurons for each subtype increases. We conclude that development of the Nematostella nerve net is stereotyped between individuals. Our data also imply that one aspect of generating adult cnidarian nervous systems is to modify the basic structural architecture generated in the juvenile by increasing neural number proportionally with size.

To determine differences in oropharyngeal squamous cell carcinoma (OPSCC) incidence between 1975 and 2014 stratified by race, sex, and age.We obtained age-adjusted OPSCC incidence rates for race and sex groups from 1975 to 2014 using the Surveillance, Epidemiology, and End Results 9 database. We defined OPSCC as cancers of the base of tongue, lingual/palatine tonsil, oropharynx, soft palate, uvula, and Waldeyer's ring. We used Joinpoint analyses to determine incidence trends for race/sex/age groupings.There were 38,624 oropharyngeal primary tumors in the analyses. Males accounted for 74% of sample population, and whites accounted for 84% of tumors. Overall, there was a 57.3% increase in incidence of oropharyngeal between 1975 and 2014. For blacks and whites, average incidence was lower for females than males. Rates for black males aged ≥50years was highest for most of the follow-up time but decreased sharply around 1988 and were surpassed by the significant increase in incidence in white males aged 50-59 (1995-2014 APC=4.07, p0.001) and ≥60years (2002-2014 APC=4.25, p0.001). For males aged ≥60, whites had higher rates than blacks starting in 2010. OPSCC incidence in White males (10.99 per 100,000 person-years) surpassed rates in Blacks (10.14 per 100,000 person-years) beginning in 2008.OPSCC has significantly increased in the United States in the last 40 years. This overall increase in OPSCC can primarily be attributed to white males. OPSCC prevention and early detection efforts could target these demographic factors to decrease rising OPSCC incidence.

A lack of longitudinal studies impedes the understanding of whether visual processing skills significantly influence reading performance. The present study assessed if multi-element processing (MEP), a visual processing task comprising only non-verbal stimuli, was predominantly related with decoding or sight-word reading. One hundred Spanish pre-reading children were evaluated on their MEP, naming speed (RAN), phonemic awareness (PA), letter knowledge (LK) and IQ. Early reading level was measured in first grade. In third grade, four reading lists consisting of short and long, high- and low-frequency words were administered. Results from path analyses revealed that, after controlling for RAN, PA, LK, IQ and early reading level, MEP was a significant predictor of the reading of long low-frequency words only. This result suggests that, in the transparent Spanish orthography, pre-reading MEP is significantly linked to future decoding skill. This is the first study to provide empirical evidence that pre-reading MEP predicts future reading.

The Golgi complex is the central unit of the secretory pathway, modifying, processing and sorting proteins and lipids to their correct cellular localisation. Changes to proteins at the Golgi complex can have deleterious effects on the function of this organelle, impeding trafficking routes through it, potentially resulting in disease. It is emerging that several Rho GTPase proteins, namely Cdc42, RhoBTB3, RhoA and RhoD are at least in part localised to the Golgi complex, and a number of studies have shown that dysregulation of their levels or activity can be associated with cellular changes which ultimately drive cancer progression. In this mini-review we highlight some of the recent work that explores links between form and function of the Golgi complex, Rho GTPases and cancer.

Summary: Alpha-1 adrenergic receptors (α1-ARs) play adaptive and protective roles in the heart. Dabuzalgron is an oral selective α1A-AR agonist that was well tolerated in multiple clinical trials of treatment for urinary incontinence, but has never been used to treat heart disease in humans or animal models. In this study, the authors administered dabuzalgron to mice treated with doxorubicin (DOX), a widely used chemotherapeutic agent with dose-limiting cardiotoxicity that can lead to heart failure (HF). Dabuzalgron protected against DOX-induced cardiotoxicity, likely by preserving mitochondrial function. These results suggest that activating cardiac α1A-ARs with dabuzalgron, a well-tolerated oral agent, might represent a novel approach to treating HF. Key Words: alpha adrenergic receptors, anthracyclines, cardioprotection, catecholamines, heart failure

Background There is limited evidence whether being on fludrocortisone prevents vasovagal syncope. Objectives The authors sought to determine whether treatment with fludrocortisone reduces the proportion of patients with recurrent vasovagal syncope by at least 40%, representing a pre-specified minimal clinically important relative risk reduction. Methods The multicenter POST 2 (Prevention of Syncope Trial 2) was a randomized, placebo-controlled, double-blind trial that assessed the effects of fludrocortisone in vasovagal syncope over a 1-year treatment period. All patients had >2 syncopal spells and a Calgary Syncope Symptom Score >−3. Patients received either fludrocortisone or matching placebo at highest tolerated doses from 0.05 mg to 0.2 mg daily. The main outcome measure was the first recurrence of syncope. Results The authors randomized 210 patients (71% female, median age 30 years) with a median 15 syncopal spells over a median of 9 years equally to fludrocortisone or placebo. Of these, 96 patients had ≥1 syncope recurrences, and only 14 patients were lost to follow-up before syncope recurrence. There was a marginally nonsignificant reduction in syncope in the fludrocortisone group (hazard ratio [HR]: 0.69: 95% confidence interval [CI]: 0.46 to 1.03; p = 0.069). In a multivariable model, fludrocortisone significantly reduced the likelihood of syncope (HR: 0.63; 95% CI: 0.42 to 0.94; p = 0.024). When the analysis was restricted to outcomes after 2 weeks of dose stabilization, there was a significant benefit due to fludrocortisone (HR: 0.51; 95% CI: 0.28 to 0.89; p = 0.019). Conclusions The study did not meet its primary objective of demonstrating that fludrocortisone reduced the likelihood of vasovagal syncope by the specified risk reduction of 40%. The study demonstrated a significant effect after dose stabilization, and there were significant findings in post hoc multivariable and on-treatment analyses. (A randomised clinical trial of fludrocortisone for the prevention of vasovagal syncope; ISRCTN51802652 ; Prevention of Syncope Trial 2 [POST 2]; NCT00118482 )

The inner tracker of the ATLAS detector is scheduled to be replaced by a completely new silicon-based inner tracker (ITk) for the Phase-II of the CERN LHC (HL-LHC). The silicon strip detector covers the volume 40 R 100 cm in the radial and | z | 300 cm in the longitudinal directions. The silicon sensors for the detector will be fabricated using the n + -on-p 6-inch wafer technology, for a total of 22,000 wafers. Intensive studies were carried out on the final prototype sensors ATLAS17LS fabricated by Hamamatsu Photonics (HPK). The charge collection properties were examined using penetrating 90Sr β -rays and the ALIBAVA fast readout system for the miniature sensors of 1 cm × 1 cm in area. The samples were irradiated by protons in the 27 MeV Birmingham Cyclotron, the 70 MeV CYRIC at Tohoku University, and the 24 GeV CERN-PS, and by neutrons at Ljubljana TRIGA reactor for fluence values up to 2 × 1015 n eq /cm 2 . The change in the charge collection with fluence was found to be similar to the previous prototype ATLAS12, and acceptable for the ITk. Sensors with two active thicknesses, 300 μ m (standard) and 240 μ m (thin), were compared and the difference in the charge collection was observed to be small for bias voltages up to 500 V. Some samples were also irradiated with gamma radiation up to 2 MGy, and the full depletion voltage was found to decrease with the dose. This was caused by the Compton electrons due to the 60Co gamma radiation. To summarize, the design of the ATLAS17LS and technology for its fabrication have been verified for implementation in the ITk. We are in the stage of sensor pre-production with the first sensors already delivered in January of 2020.

Objective/Hypothesis Early diagnosis and stage at presentation, two prognostic factors for survival among patients with head and neck cancer (HNC), are significantly impacted by a patient’s health insurance status. We aimed to assess the impact of the Patient Protection and Affordable Care Act (ACA) on stage at presentation across socioeconomic and demographic subpopulations of HNC patients in the United States. Study Design Retrospective data analysis. Methods The National Cancer Database, a hospital-based cancer database (2011–2015), was queried for adults aged 18–64 years and diagnosed with a malignant primary HNC. The outcome of interest was change in early-stage diagnoses between 2011–2013 (pre-ACA) and 2014–2015 (post-ACA) using logistic regression models. Results A total of 91,137 HNC cases were identified in the pre-ACA (n = 53,726) and post-ACA (n = 37,411) years. Overall, the odds of early-stage diagnoses did not change significantly post-ACA (aOR = 0.97, 95 % CI 0.94, 1.00; p = 0.081). However, based on health insurance status, HNC patients with Medicaid were significantly more likely to present with early-stage disease post-ACA (aOR = 1.12, 95 % CI 1.03, 1.21; p = 0.007). We did not observe increased odds of early-stage presentation for other insurance types. Males were less likely to present with early-stage disease, pre- or post-ACA. Conclusions We demonstrate a significant association between ACA implementation and increased early-stage presentation among Medicaid-enrolled HNC patients. This suggests that coverage expansions through the ACA may be associated with increased access to care and may yield greater benefits among low-income HNC patients.

High-content analysis (HCA) provides quantitative multiparametric cellular fluorescence data. From its origins in discovery toxicology, it is now addressing fundamental questions in drug delivery. Nanoparticles (NPs), polymers, and intestinal permeation enhancers are being harnessed in drug delivery systems to modulate plasma membrane properties and the intracellular environment. Identifying comparative mechanistic cytotoxicity on sublethal events is crucial to expedite the development of such systems. NP uptake and intracellular routing pathways are also being dissected using chemical and genetic perturbations, with the potential to assess the intracellular fate of targeted and untargeted particles in vitro. As we discuss here, HCA is set to make a major impact in preclinical delivery research by elucidating the intracellular pathways of NPs and the in vitro mechanistic-based toxicology of formulation constituents. Department of Agriculture, Food and the Marine European Commission - Seventh Framework Programme (FP7) Science Foundation Ireland

A method has been developed for the preparation of amines through a process of coupling acyl succinimides derived from commercially available carboxylic acids with amines to afford the corresponding amides. These amides are then reduced in situ with either diisobutylaluminum hydride or lithium aluminum hydride. The reaction tandem of the coupling reaction followed by the reduction affords the amine in fair to good yields after purification by flash chromatography. This one-pot, two reaction tandem process has been successfully applied to the synthesis of the calcimimetic agents cinacalcet, NPS R-467, and NPS R-568.

Ethnic-specific differences in minor allele frequency impact variant categorization for genetic screening of nonsyndromic hearing loss (NSHL) and other genetic disorders. We sought to evaluate all previously reported pathogenic NSHL variants in the context of a large number of controls from ethnically distinct populations sequenced with orthogonal massively parallel sequencing methods. We used HGMD, ClinVar, and dbSNP to generate a comprehensive list of reported pathogenic NSHL variants and re-evaluated these variants in the context of 8,595 individuals from 12 populations and 6 ethnically distinct major human evolutionary phylogenetic groups from three sources (Exome Variant Server, 1000 Genomes project, and a control set of individuals created for this study, the OtoDB). Of the 2,197 reported pathogenic deafness variants, 325 (14.8%) were present in at least one of the 8,595 controls, indicating a minor allele frequency (MAF) > 0.00006. MAFs ranged as high as 0.72, a level incompatible with pathogenicity for a fully penetrant disease like NSHL. Based on these data, we established MAF thresholds of 0.005 for autosomal-recessive variants (excluding specific variants in GJB2) and 0.0005 for autosomal-dominant variants. Using these thresholds, we recategorized 93 (4.2%) of reported pathogenic variants as benign. Our data show that evaluation of reported pathogenic deafness variants using variant MAFs from multiple distinct ethnicities and sequenced by orthogonal methods provides a powerful filter for determining pathogenicity. The proposed MAF thresholds will facilitate clinical interpretation of variants identified in genetic testing for NSHL. All data are publicly available to facilitate interpretation of genetic variants causing deafness.

Using transition metals such as manganese(II), iron(II), cobalt(II), nickel(II), copper(II), and zinc(II), several new metal complexes of cross-bridged tetraazamacrocyclic chelators namely, cyclen- and cyclam-analogs with benzyl groups, were synthesized and screened for in vitro antimalarial activity against chloroquine-resistant (W2) and chloroquine-sensitive (D6) strains of Plasmodium falciparum. The metal-free chelators tested showed little or no antimalarial activity. All the metal complexes of the dibenzyl cross-bridged cyclam ligand exhibited potent antimalarial activity. The Mn(2+) complex of this ligand was the most potent with IC50s of 0.127 and 0.157μM against the chloroquine-sensitive (D6) and chloroquine-resistant (W2) P. falciparum strains, respectively. In general, the dibenzyl hydrophobic ligands showed better anti-malarial activity compared to the activity of monobenzyl ligands, potentially because of their higher lipophilicity and thus better cell penetration ability. The higher antimalarial activity displayed by the manganese complex for the cyclam ligand in comparison to that of the cyclen, correlates with the larger pocket of cyclam compared to that of cyclen which produces a more stable complex with the Mn(2+). Few of the Cu(2+) and Fe(2+) complexes also showed improvement in activity but Ni(2+), Co(2+) and Zn(2+) complexes did not show any improvement in activity upon the metal-free ligands for anti-malarial development.

Purpose People with seizure disorders experience elevated rates of psychopathology, often undiagnosed and untreated. Accurate diagnosis of psychopathology remains an important goal of quality health care for people with seizure disorders. One of the most widely used dimensional measures of psychopathology is the Minnesota Multiphasic Personality Inventory—Second Edition (MMPI-2). Research in heterogeneous mental health samples suggests that the 2008 revision of this measure, the Minnesota Multiphasic Personality Inventory—Second Edition—Restructured Form (MMPI-2-RF), offers better construct fidelity and more cost-effective administration. This study seeks to extend research on MMPI-2-RF scale elevations to a sample of people with seizure disorders. Methods In a consecutive, heterogeneous sample of people with seizure disorders, MMPI-2 and MMPI-2-RF scores were compared in terms of categorical classification agreement (clinically elevated versus not clinically elevated). Scores were also compared in terms of variance attributable to diagnosis-specific items, general demoralization, subtle items, social desirability, and demographic factors. Key findings Scores on MMPI-2 and MMPI-2-RF provided a statistically significant level of agreement between corresponding clinical diagnostic scales ranging from 68% to 84%. Most classification disagreement was attributable to MMPI-2 clinical scale elevations when MMPI-2-RF scales were not elevated. Regression analysis supported the interpretation that general demoralization, subtle items, social desirability, and demographic factors led to MMPI-2 clinical scale elevations. Significance The results provide evidence that in the context of strong psychopathology classification agreement, the MMPI-2-RF restructured clinical scales provide better construct fidelity compared with the more trait heterogeneous MMPI-2 clinical scales. These results should encourage clinicians to use the MMPI-2 Restructured Form (MMPI-2-RF) for improved psychopathology assessment compared with the MMPI-2 in patients with seizure disorders.

Enlarged left ventricle, aortic root dilation, and obstructive sleep apnea (OSA) have been reported as findings in retired professional football players. However, there are few studies examining the relationship between OSA risk and left ventricular hypertrophy (LVH) or aortic root dilation in this

Summary Floods with differing sizes and durations are likely to impact riparian systems in different hydrologic and geochemical ways. Here the impact of flood size and duration was investigated. Flood-driven recharge along predominantly losing reaches of the Bill Williams River (western Arizona, USA) later reemerges as baseflow in downstream gaining reaches, and the river’s longest losing reach (Planet Valley) retains and releases the most flood recharge. River discharge volume and flow intermittency downstream of Planet Valley is highly dependent upon the length of time since the last major flood. After large floods (e.g. 2004–2005), baseflow was dominated by the flood’s chemical (SO 4 , Cl) and isotopic (δ 18 O H2O , δ 2 H H2O , δ 34 S SO4 , δ 18 O SO4 ) composition for long periods (>4 years), suggesting that the largest events result in much more flood recharge and a longer persistence of floodwater in the subsurface than after smaller, more recent events. The continued dominance of baseflow by 2004–2005 floodwater nearly 5 years later—despite three smaller floods in 2007, 2008 and 2009—highlights the long-term impacts that the largest floods have on riparian water composition. Of these three recent events, only the largest and longest (in 2009) caused observable changes in both baseflow volume and the composition of baseflow and riparian groundwater, thereby suggesting that a threshold of flood size and duration exists for floods to alter the system’s state and behavior. The river’s dependence on large winter floods and the tendency of a particular set of atmospheric conditions (associated with El Nino-Southern Oscillation, or ENSO) to cause the region’s largest winter floods indicates the importance of ENSO to the system, and that future changes to ENSO caused by climate change could drastically alter the flood properties and overall hydrology of southwestern rivers.

Rationale: Gq signaling in cardiac myocytes is classically considered toxic. Targeting Gq directly to test this is problematic, because cardiac myocytes have many Gq-coupled receptors. Objective: Test whether Gq coupling is required for the cardioprotective effects of an alpha-1A-AR (adrenergic receptor) agonist. Methods and Results: In recombinant cells, a mouse alpha-1A-AR with a 6-residue substitution in the third intracellular loop does not couple to Gq signaling. Here we studied a knockin mouse with this alpha-1A-AR mutation. Heart alpha-1A receptor levels and antagonist affinity in the knockin were identical to wild-type. In wild-type cardiac myocytes, the selective alpha-1A agonist A61603-stimulated phosphoinositide-phospholipase C and myocyte contraction. In myocytes with the alpha-1A knockin, both A61603 effects were absent, indicating that Gq coupling was absent. Surprisingly, A61603 activation of cardioprotective ERK (extracellular signal-regulated kinase) was markedly impaired in the KI mutant myocytes, and A61603 did not protect mutant myocytes from doxorubicin toxicity in vitro. Similarly, mice with the α1A KI mutation had increased mortality after transverse aortic constriction, and A61603 did not rescue cardiac function in mice with the Gq coupling-defective alpha-1A receptor. Conclusions: Gq coupling is required for cardioprotection by an alpha-1A-AR agonist. Gq signaling can be adaptive.

Computer access technology (CAT) allows people who have trouble using a standard computer keyboard, mouse, or monitor to access a computer. CAT is critical for enhancing the educational and vocational opportunities of people with disabilities. Choosing the most appropriate CAT is a collaborative decision-making process involving the consumer, clinician(s), and third party payers. The challenges involved and potential technological solutions are discussed.

Offset schemes are advocated as a way that continued development and environmental restoration can be achieved concurrently. We used a simple modelling approach to evaluate proposed offsets schemes, with scenarios that required offsetting the impacts of clearing woodlands either by revegetation of cleared land or by improving the habitat value of degraded woodland. Each simulation used the attribute table of a single GIS polygon layer to obtain data and record results. We investigated the likely consequences of these schemes for three groups of species with different foraging resource requirements: shrub-dependent; canopy-dependent and old tree-dependent. Only the shrub-dependent group, whose requirements could be rapidly grown, saw increases in suitable habitat in the landscape within our 30 year simulations. The habitat of the canopy-dependent group initially declined but began to increase towards the end of some simulations, while the old tree-dependent group’s habitat declined. When a simple measure of spatial configuration was considered further differences between the schemes were highlighted. The simulation results demonstrate that assessing only the eventual benefit score of a scheme can hide the losses sustained by some elements of biodiversity. We recommend this type of simple modelling approach as the first step in determining whether a proposed offsets scheme is worth investigating further. In particular, the ability to represent the scheme’s predicted consequences as maps and graphs assists decision makers in judging whether the scheme has sufficient merit to warrant a full assessment and subsequent implementation; or needs some adjustments to achieve its aims, or is seriously flawed.

Increasing our knowledge of the secretory pathway remains fundamental to a greater understanding of cell health and disease. High throughput cell biology approaches provide a new opportunity to study this important process at a truly genome-wide scale. This review highlights how techniques such as high throughput subcellular localisation, RNA interference screening and automated microscopy are now being harnessed to provide new information about secretory pathway function.

The pace and magnitude of biodiversity loss has led to wide recognition that efforts to conserve individual species must be complemented by assessment and planning at community and ecosystem levels. Emerging protocols for assessing the conservation status of communities include as central criteria the current extent, historical reduction and contemporary rate of decline in geographic distribution. Estimation of these parameters is confronted by methodological challenges, data limitations and uncertainties that may vary from case to case. We describe an approach to these issues comprising five steps: (1) classification of the community using an analysis of data from systematic ground surveys; (2) mapping to produce a contemporary baseline distribution (1980s); (3) interpolation to produce a historical distribution (pre-European settlement); (4) interpretation of satellite imagery to update the distribution (various dates up to 2004) and (5) assessment of change in extent over historical and contemporary time scales incorporating plausible bounds of uncertainty around best estimates. The bounds can be based on areas for which image interpretation produces uncertain diagnosis of clearing and differences between credible alternative base maps of the same area. We demonstrated the approach using a case study of Coolibah–Black Box Woodland, a declining semi-arid woodland community in Australia and found that 61% (plausible range 50–67%) of the woodland community had been cleared since European settlement and that during 1998–2004 the community continued to decline on average by 135.3(±21.7) km 2 each year, or 1.7(±0.3)% of each previous year’s distribution, apparently accelerating in recent years. Strengths of the approach include the use of biological data (cf. remote sensing) to distinguish the target assemblage from others, the use of historical and contemporary base lines to examine change over different time scales, and the use of bounded estimates to incorporate uncertainty into the assessment.

The study examined changes in visual attention in schizophrenia following training with a social-cognitive remediation package designed to improve facial emotion recognition (the Micro-Expression Training Tool; METT). Forty out-patients with schizophrenia were randomly allocated to active training (METT; n = 26), or repeated exposure (RE; n = 14); all completed an emotion recognition task with concurrent eye movement recording. Emotion recognition accuracy was significantly improved in the METT group, and this effect was maintained after one week. Immediately following training, the METT group directed more eye movements within feature areas of faces (i.e., eyes, nose, mouth) compared to the RE group. The number of fixations directed to feature areas of faces was positively associated with emotion recognition accuracy prior to training. After one week, the differences between METT and RE groups in viewing feature areas of faces were reduced to trends. However, within group analyses of the METT group revealed significantly increased number of fixations to, and dwell time within, feature areas following training which were maintained after one week. These results provide the first evidence that improvements in emotion recognition following METT training are associated with changes in visual attention to the feature areas of emotional faces. These findings support the contribution of visual attention abnormalities to emotion recognition impairment in schizophrenia, and suggest that one mechanism for improving emotion recognition involves re-directing visual attention to relevant features of emotional faces.

The Gawler Range Volcanics (GRV) of South Australia are an intracontinental, subaerial Large Igneous Province (LIP) formed during supercontinent assembly in the Mesoproterozoic. Like many LIPs, the GRV are intraplate, were erupted over a geologically short time ( 2 million years), and are dominated by voluminous lavas. However, the GRV are overwhelmingly dominated by felsic units. The GRV are essentially undeformed and have not been deeply buried so that their original textures are exceptionally well preserved. Furthermore, they are well exposed in very gently northward and eastward dipping sections up to 4 km thick, allowing assessment of the facies architecture and evolution of this felsic volcanic LIP. The evolution of the GRV can be clearly separated into two main stages. Initial eruptions at numerous volcanic centres produced small to moderate volume, geochemically distinct, felsic lavas and lava domes, together with ignimbrites and minor mafic and intermediate lavas, forming a sequence 0.5 to 3 km thick. Volcanic activity in this lower sequence varied from effusive to explosive and was not much different in style or products from Phanerozoic felsic volcanic provinces. However, the second stage produced at least three voluminous felsic units, each of which represents about 1000–3000 km3 of magma. Moreover, the dominance of evenly porphyritic textures and lack of pyroclastic textures (pumice, shards, broken crystal, lithic clasts) in these units suggests that they were erupted effusively and flowed as lavas. Each of these felsic lavas are generally dominated by a single uniform composition, and commonly mingled with a subordinate and compositionally distinct lava.

White cypress pine Callitris glaucophylla is an important native tree widespread in heavily cleared savanna woodlands of the south-east Australian wheat-sheep belt. C. glaucophylla woodlands have been the focus of vigorous debate regarding their structure and dynamics during Aboriginal times and following European settlement, and the causes of structural changes are poorly understood. Management of contemporary woodland remnants is therefore controversial, and has so far lacked a predictive, process-based approach. We sought to determine whether a simple process model with recruitment, growth and survival mediated by rainfall and competition, could accurately simulate medium-term dynamics of C. glaucophylla woodlands in which fire has typically been excluded. Model parameters were optimised using reference data from 6- to 36-year silvicultural trials in central-western New South Wales (NSW). Predictions of the calibrated model compared favourably to actual growth, survival and recruitment of C. glaucophylla in the reference data, as well as in an independent dataset not used in optimisation. We tested whether remaining differences between predictions and actual data were related to variation in site productivity (a function of moisture availability as influenced by soil and landscape factors not modelled). Growth was overestimated in low-productivity sites and underestimated in high-productivity sites. Growth predictions were improved by using site-specific expected maximum heights to reflect site productivity. We also tested whether the model and parameter set produce dynamics consistent with those observed over the last half century. Modelled growth, survival, recruitment and competition processes were consistent with data and observations in previous studies on historical dynamics and stand behaviour of C. glaucophylla. The model has a wide range of potential applications for understanding past and predicting future stand dynamics. To illustrate an application of the model to thinning, a strategy currently being implemented to manage tree densities in inland NSW, we compared predicted stand structures in simulations of different thinning treatments. The simulations showed that without careful manipulation of existing canopy cover, stands can re-fill to pre-thinning densities within relatively short periods of time.

The role of membrane traffic is to transfer cargo between distinct subcellular compartments. Each individual trafficking event involves the creation, transport and fusion of vesicular and tubular carriers that are formed and regulated via cytoplasmic coat protein complexes. The dynamic nature of this process is therefore highly suitable for studying using live cell imaging techniques. Although these approaches have raised further questions for the field, they have also been instrumental in providing essential new information, in particular relating to the morphology of transport carriers and the exchange kinetics of coat proteins and their regulators on membranes. Here, we present an overview of live cell-imaging experiments that have been used in the study of coated-vesicle transport, and provide specific examples of their impact on our understanding of coat function.

Integrative approaches to study protein function in a cellular context are a vital aspect of understanding human disease. Genome sequencing projects provide the basic catalogue of information with which to unravel gene function, but more systematic applications of this resource are now necessary. Here, we describe and test a platform with which it is possible to rapidly use RNA interference in cultured mammalian cells to probe for proteins involved in constitutive protein secretion. Synthetic small interfering RNA molecules are arrayed in chambered slides, then incubated with cells and an assay for secretion performed. Automated microscopy is used to acquire images from the experiments, and automated single-cell analysis rapidly provides reliable quantitative data. In test arrays of 92 siRNA spots targeting 37 prospective membrane traffic proteins, our approach identifies 7 of these as being important for the correct delivery of a secretion marker to the cell surface. Correlating these findings with other screens and bioinformatic information makes these candidates highly likely to be novel membrane traffic machinery components.

Objective: The aim of this study was to examine published reports for sources of excessive variance in neuroleptic malignant syndrome (NMS) incidence estimates. Data Sources: An unrestricted computerized MEDLINE search was conducted with a comprehensive search logic and supplemented by secondary references and a manual search of an extensive personal library. Study Selection: Studies were analyzed if they presented original data and provided at least 2 of the following: number of NMS cases, number of patients at risk, or ratio of cases to patients at risk. Twenty-six of the 28 candidate studies met these minimal criteria. Data Extraction: Variables included incidence, year of study publication, mean year of NMS occurrence, patient population at risk, study design, diagnostic criteria, and country of origin. Data Synthesis: Standard error, which reflects study size, accounted for 90.8% of the variance (β = .953, P < .001) in this international series of 26 NMS incidence estimates. Incidence was significantly lower in 7 studies the time end points of which were set in advance of case identification (χ2 = 71.08, P ≪ .001). No other variable was significantly related to incidence. Conclusions: Neuroleptic malignant syndrome incidence estimates to date are non–trivially biased such that larger study size (patients at risk) is strongly related to lower observed incidence. Future studies can minimize the contribution of this and other sources of experimental error by incorporating several very feasible recommendations.

Discrete hollows in the bar tops of the South Saskatchewan River are described that form a newly-recognized morphological element of sandy braided rivers. These bar-top hollows, which are up to 1.7 m deep and may extend for 10–30 m down and across flow, have a circular to ovoid planform and are shown, through use of ground penetrating radar, to be filled by a series of distinct, often angle-of-repose, foresets. The hollows form by both erosion and bar-top deposition and may be generated by bar-tail accretion, cross-bar channel cutoff and subsequent fill or lateral accretion at the bar-head. Bar-top hollows occur in the upper part of the bar depositional sequence and may thus prove useful indicators for braid bar reconstruction in ancient sediments, and should not be confused with channel scour.

Adrenergic receptors (ARs) play an important role in the regulation of cardiac function. Cardiac inotropy is primarily regulated by beta(1)-ARs. However, alpha(1)-ARs may play an important role in inotropy during heart failure. Previous work has suggested that the alpha(1B)-AR modulates beta(1)-AR function in the heart. The potential role of the alpha(1A)-AR has not been previously studied. We used transgenic mice that express constitutively active mutant (CAM) forms of the alpha(1A)-AR or alpha(1B)-AR regulated by their endogenous promoters. Expression of the CAM alpha(1A)-AR or CAM alpha(1B)-AR had no effect on basal cardiac function (developed pressure, +dP/dT, -dP/dT, heart rate, flow rate). However, both alpha(1)-AR subtypes significantly decreased isoproterenol-stimulated +dP/dT. Pertussis toxin had no effect on +dP/dT in CAM alpha(1A)-AR hearts but restored +dP/dT to non-transgenic values in CAM alpha(1B)-AR hearts. Radioligand binding indicated a selective decrease in the density of beta(1)-ARs in both CAM mice. However, G-proteins, cAMP, or the percentage of high and low affinity states were unchanged in either transgenic compared with control. These data demonstrate that CAM alpha(1A)- and alpha(1B)-ARs both down regulate beta(1)-AR-mediated inotropy in the mouse heart. However, alpha(1)-AR subtypes are coupled to different beta-AR mediated signaling pathways with the alpha(1B)-AR being pertussis toxin sensitive.

We present experimental results on a technique for alloy composition grading within distributed Bragg reflectors (DBRs) for use in vertical-cavity, surface-emitting lasers (VCSELs). This technique, implemented using molecular beam epitaxy (MBE), employs multiple group-III sources, with different fluxes, to realize a set of Al x Ga 1− x As compositions without changing cell temperatures. By varying the source combination and layer thickness, different functional forms of compositional grading, such as linear and parabolic, can be closely approximated. This method, in combination with modulated doping profiles, was used to grow 980 nm VCSELs with low threshold currents, high output powers, and threshold voltages less than 1.45 V.

This paper investigates the effect of player preparation, ground conditions and weather conditions upon the injury risk for Rugby Union players. A population-based case-control study was performed using a sample ( n =1043) of New Zealand Rugby Union players aged 16 y and above. Details concerning game preparation (warm-up and usual position), and ground and weather conditions (precipitation, wind and temperature) were obtained from the players. If players were injured during the season ( n =624) they were asked to provide details about the game in which they were last injured. Uninjured players ( n =419) provided details about the last game in which they played. Injuries were more likely to occur when games were played on hard grounds or in calm or warm conditions. Playing out of position and the duration of warming up did not significantly alter the risk of injury. When player preparation, ground and weather conditions, grade, age, playing position and rugby experience were simultaneously controlled for, hard ground and the absence of wind were associated with increased risk. The influence of these factors may be indirect, through adaptation to the conditions in which a game is played.

Background Migraine is a common neurological disorder affecting children, in which the headache is often preceded or accompanied by a complex of neurological symptoms known as an aura. Persistent visual symptoms are rare, with typical visual aura sometimes being poorly distinguished from other visual disturbances. Methods We describe the case of a 12-year-old girl who has experienced persistent, constant symptoms throughout the visual fields of white, bright, jagged spots and black and white flashes with sparkles and dots since May 2010. She also has palinopsia, squiggles, and photophobia. The child's drawing of her visual symptoms helps illustrate the case and illuminate her ordeal. Results The child's visual symptoms have so far been resistant to pharmacological therapy. Conclusion Further insight is needed into this debilitating condition to allow effective management in the pediatric population.