SIRT6 Is Responsible for More Efficient DNA Double-Strand Break Repair in Long-Lived Species.

DNA repair has been hypothesized to be a longevity determinant, but the evidence for it is based largely on accelerated aging phenotypes of DNA repair mutants. Here, using a panel of 18 rodent species with diverse lifespans, we show that more robust DNA double-strand break (DSB) repair, but not nucleotide excision repair (NER), coevolves with longevity. Evolution of NER, unlike DSB, is shaped primarily by sunlight exposure. We further show that the capacity of the SIRT6 protein to promote DSB repair accounts for a major part of the variation in DSB repair efficacy between short- and long-lived species. We dissected the molecular differences between a weak (mouse) and a strong (beaver) SIRT6 protein and identified five amino acid residues that are fully responsible for their differential activities. Our findings demonstrate that DSB repair and SIRT6 have been optimized during the evolution of longevity, which provides new targets for anti-aging interventions.

Representative cancer-associated U2AF2 mutations alter RNA interactions and splicing.

High-throughput sequencing of hematologic malignancies and other cancers has revealed recurrent mis-sense mutations of genes encoding pre-mRNA splicing factors. The essential splicing factor U2AF2 recognizes a polypyrimidine-tract splice-site signal and initiates spliceosome assembly. Here, we investigate representative, acquired U2AF2 mutations, namely N196K or G301D amino acid substitutions associated with leukemia or solid tumors, respectively. We determined crystal structures of the wild-type (WT) compared with N196K- or G301D-substituted U2AF2 proteins, each bound to a prototypical AdML polypyrimidine tract, at 1.5, 1.4, or 1.7 Å resolutions. The N196K residue appears to stabilize the open conformation of U2AF2 with an inter-RNA recognition motif hydrogen bond, in agreement with an increased apparent RNA-binding affinity of the N196K-substituted protein. The G301D residue remains in a similar position as the WT residue, where unfavorable proximity to the RNA phosphodiester could explain the decreased RNA-binding affinity of the G301D-substituted protein. We found that expression of the G301D-substituted U2AF2 protein reduces splicing of a minigene transcript carrying prototypical splice sites. We further show that expression of either N196K- or G301D-substituted U2AF2 can subtly alter splicing of representative endogenous transcripts, despite the presence of endogenous, WT U2AF2 such as would be present in cancer cells. Altogether, our results demonstrate that acquired U2AF2 mutations such as N196K and G301D are capable of dysregulating gene expression for neoplastic transformation.

Genetic analyses of the inheritance and expressivity of autonomous endosperm formation in Hieracium with different modes of embryo sac and seed formation.

Background and Aims: Apomixis, or asexual seed formation, in polyploid Hieracium subgenus Pilosella species results in clonal progeny with a maternal genotype. An aposporous embryo sac forms mitotically from a somatic cell, without prior meiosis, while embryo and endosperm formation is fertilization independent (autonomous). The latter two developmental components are tightly linked in Hieracium . Recently, two plants, AutE196 and AutE24, were identified from two different crosses. Both form embryo sacs via the sexual route by undergoing meiosis, and embryo development requires fertilization; however, 18 % of embryo sacs can undergo autonomous endosperm (AutE) formation. This study investigated the qualitative and quantitative inheritance of the AutE trait and factors influencing phenotype expressivity. An additional focus was to identify the linkage group bearing the AutE locus in AutE196. Methods: Crosses and cytology were used to examine the inheritance of AutE from AutE24 and AutE196, and to reintroduce apomictic components into AutE plants, thereby changing the ploidy of developing embryo sacs and increasing the dosage of AutE loci. Markers from a Hieracium apomict linkage map were examined within a backcrossed AutE196 mapping population to identify the linkage group containing the AutE196 locus. Key Results: Qualitative autonomous endosperm in the AutE24 line was conferred by a single dominant locus, and the trait was transmitted through male and female gametes in AutE196 and AutE24. Expressivity of the trait did not significantly increase when AutE loci from AutE196 and AutE24 were both present in the progeny, within embryo sacs formed via apospory, or sexually derived embryo sacs with increased ploidy. It remains unclear if these are identical loci. Conclusions: The qualitative trait of autonomous endosperm formation is conferred by single dominant loci in AutE196 and AutE24. High expressivity of autonomous endosperm formation observed in apomicts requires additional genetic factors. Potential candidates may be signals arising from fertilization-independent embryo formation.

Generation of an integrated Hieracium genomic and transcriptomic resource enables exploration of small RNA pathways during apomixis initiation.

BACKGROUND: Application of apomixis, or asexual seed formation, in crop breeding would allow rapid fixation of complex traits, economizing improved crop delivery. Identification of apomixis genes is confounded by the polyploid nature, high genome complexity and lack of genomic sequence integration with reproductive tissue transcriptomes in most apomicts. RESULTS: A genomic and transcriptomic resource was developed for Hieracium subgenus Pilosella (Asteraceae) which incorporates characterized sexual, apomictic and mutant apomict plants exhibiting reversion to sexual reproduction. Apomicts develop additional female gametogenic cells that suppress the sexual pathway in ovules. Disrupting small RNA pathways in sexual Arabidopsis also induces extra female gametogenic cells; therefore, the resource was used to examine if changes in small RNA pathways correlate with apomixis initiation. An initial characterization of small RNA pathway genes within Hieracium was undertaken, and ovary-expressed ARGONAUTE genes were identified and cloned. Comparisons of whole ovary transcriptomes from mutant apomicts, relative to the parental apomict, revealed that differentially expressed genes were enriched for processes involved in small RNA biogenesis and chromatin silencing. Small RNA profiles within mutant ovaries did not reveal large-scale alterations in composition or length distributions; however, a small number of differentially expressed, putative small RNA targets were identified. CONCLUSIONS: The established Hieracium resource represents a substantial contribution towards the investigation of early sexual and apomictic female gamete development, and the generation of new candidate genes and markers. Observed changes in small RNA targets and biogenesis pathways within sexual and apomictic ovaries will underlie future functional research into apomixis initiation in Hieracium.

A reference genetic linkage map of apomictic Hieracium species based on expressed markers derived from developing ovule transcripts.

BACKGROUND AND AIMS: Apomixis in plants generates clonal progeny with a maternal genotype through asexual seed formation. Hieracium subgenus Pilosella (Asteraceae) contains polyploid, highly heterozygous apomictic and sexual species. Within apomictic Hieracium, dominant genetic loci independently regulate the qualitative developmental components of apomixis. In H. praealtum, LOSS OF APOMEIOSIS (LOA) enables formation of embryo sacs without meiosis and LOSS OF PARTHENOGENESIS (LOP) enables fertilization-independent seed formation. A locus required for fertilization-independent endosperm formation (AutE) has been identified in H. piloselloides. Additional quantitative loci appear to influence the penetrance of the qualitative loci, although the controlling genes remain unknown. This study aimed to develop the first genetic linkage maps for sexual and apomictic Hieracium species using simple sequence repeat (SSR) markers derived from expressed transcripts within the developing ovaries. METHODS: RNA from microdissected Hieracium ovule cell types and ovaries was sequenced and SSRs were identified. Two different F1 mapping populations were created to overcome difficulties associated with genome complexity and asexual reproduction. SSR markers were analysed within each mapping population to generate draft linkage maps for apomictic and sexual Hieracium species. KEY RESULTS: A collection of 14 684 Hieracium expressed SSR markers were developed and linkage maps were constructed for Hieracium species using a subset of the SSR markers. Both the LOA and LOP loci were successfully assigned to linkage groups; however, AutE could not be mapped using the current populations. Comparisons with lettuce (Lactuca sativa) revealed partial macrosynteny between the two Asteraceae species. CONCLUSIONS: A collection of SSR markers and draft linkage maps were developed for two apomictic and one sexual Hieracium species. These maps will support cloning of controlling genes at LOA and LOP loci in Hieracium and should also assist with identification of quantitative loci that affect the expressivity of apomixis. Future work will focus on mapping AutE using alternative populations.

The LOSS OF APOMEIOSIS (LOA) locus in Hieracium praealtum can function independently of the associated large-scale repetitive chromosomal structure.

Apomixis or asexual seed formation in Hieracium praealtum (Asteraceae) is controlled by two independent dominant loci. One of these, the LOSS OF APOMEIOSIS (LOA) locus, controls apomixis initiation, mitotic embryo sac formation (apospory) and suppression of the sexual pathway. The LOA locus is found near the end of a hemizygous chromosome surrounded by extensive repeats extending along the chromosome arm. Similar apomixis-carrying chromosome structures have been found in some apomictic grasses, suggesting that the extensive repetitive sequences may be functionally relevant to apomixis. Fluorescence in situ hybridization (FISH) was used to examine chromosomes of apomeiosis deletion mutants and rare recombinants in the critical LOA region arising from a cross between sexual Hieracium pilosella and apomictic H. praealtum. The combined analyses of aposporous and nonaposporous recombinant progeny and chromosomal karyotypes were used to determine that the functional LOA locus can be genetically separated from the very extensive repeat regions found on the LOA-carrying chromosome. The large-scale repetitive sequences associated with the LOA locus in H. praealtum are not essential for apospory or suppression of sexual megasporogenesis (female meiosis).

Comprehensive chromatographic assessment of forced degraded in vitro transcribed mRNA.

Heightened interest in messenger RNA (mRNA) therapeutics has accelerated the need for analytical methodologies that facilitate the production of supplies for clinical trials. Forced degradation studies are routinely conducted to provide an understanding of potential weak spots in the molecule that are exploited by stresses encountered during bulk purification, production, shipment, and storage. Consequently, temperature fluctuations and excursions are often experienced during these unit operations and may accelerate mRNA degradation. Here, we present a concise panel of chromatography-based stability-indicating assays for evaluating thermally stressed in vitro transcribed (IVT) mRNA as part of a forced degradation study. We found that addition of EDTA to the mRNAs prior to heat exposure reduced the extent of degradation, suggesting that transcripts may be fragmenting via a divalent metal-ion mediated pathway. Trace divalent metal contamination that can accelerate RNA instability is likely carried over from upstream steps. We demonstrate the application of these methods to evaluate the critical quality attributes (CQAs) of mRNAs as well as to detect intrinsic process- and product-related impurities.

A single-nucleotide resolution capillary gel electrophoresis workflow for poly(A) tail characterization in the development of mRNA therapeutics and vaccines.

The 3' poly(A) tail is an important component of messenger RNA (mRNA). The length of the poly(A) tail has direct impact on the stability and translation efficiency of the mRNA molecule and is therefore considered to be a critical quality attribute (CQA) of mRNA-based therapeutics and vaccines. Various analytical methods have been developed to monitor this CQA. Methods like ion-pair reversed-phase liquid chromatography (IPRP-LC) can be used to quantify the percentage of mRNA with poly(A) tail but fail to provide further information on the actual length of poly(A). High-resolution methods such as liquid chromatography coupled with mass spectrometry (LC-MS) or next generation sequencing (NGS) can separate poly(A) tail length by one nucleotide (n/n + 1 resolution) but are complicated to implement for release testing of manufactured mRNA. In this study, a workflow utilizing capillary gel electrophoresis (CGE) for characterizing the poly(A) tail length of mRNA was developed. The CGE method demonstrated resolution comparable with the LC-MS method. With UV detection and the addition of poly(A) length markers, this method can provide poly(A) tail length information and can also provide quantitation of each poly(A) length, making it a suitable release method to monitor the CQA of poly(A) tail length.

Seasonal variation in North American level crossing crash rates is due to weather, not day length.

OBJECTIVE: Collisions between road vehicles and trains at level (grade) crossings can be devastating. Injury and economic considerations make prevention efforts of significant interest to society at all levels, and raise important safety concerns. Improving our understanding of the nature and pattern of crashes at level crossings can help inform a variety of types of safety mitigation strategies, including public education, crossing equipment and vehicle design efforts. METHOD: To this end, a database search of Canadian level crossing crashes for the 11-year period between 2007 and 2017 was conducted to confirm a previously identified seasonal variation in the frequency of level crossing crashes. To determine whether the observed winter increase in crashes was due primarily to winter reductions in light levels/day length or to other seasonal weather factors, a subsequent comparison of Canadian data to American Federal Railroad Administration (FRA) crash data was carried out. A separate inferential log-linear model analysis, using season, time of day and crossing protection type, was also used to explore the increase. RESULTS: As expected, the average rate of crossing collisions in Canada increased during winter months compared to non-winter months. While the seasonal pattern was evident in those U.S. states that experience significant changes in weather patterns (i.e., northern states), it was almost completely absent in those states that do not (i.e., southern states). The log-linear model analysis confirmed these findings. CONCLUSIONS: The seasonal variation in North American level crossing crash rate is a result of winter weather conditions, rather than differences in light levels. Future research is planned that will elucidate the specific environmental and human factors contributing to the increase.

Opportunistic tangible user interfaces for augmented reality.

Opportunistic Controls are a class of user interaction techniques that we have developed for augmented reality (AR) applications to support gesturing on, and receiving feedback from, otherwise unused affordances already present in the domain environment. By leveraging characteristics of these affordances to provide passive haptics that ease gesture input, Opportunistic Controls simplify gesture recognition, and provide tangible feedback to the user. In this approach, 3D widgets are tightly coupled with affordances to provide visual feedback and hints about the functionality of the control. For example, a set of buttons can be mapped to existing tactile features on domain objects. We describe examples of Opportunistic Controls that we have designed and implemented using optical marker tracking, combined with appearance-based gesture recognition. We present the results of two user studies. In the first, participants performed a simulated maintenance inspection of an aircraft engine using a set of virtual buttons implemented both as Opportunistic Controls and using simpler passive haptics. Opportunistic Controls allowed participants to complete their tasks significantly faster and were preferred over the baseline technique. In the second, participants proposed and demonstrated user interfaces incorporating Opportunistic Controls for two domains, allowing us to gain additional insights into how user interfaces featuring Opportunistic Controls might be designed.

Efficient CRISPR/Cas9-Mediated Knockout of an Endogenous PHYTOENE DESATURASE Gene in T1 Progeny of Apomictic Hieracium Enables New Strategies for Apomixis Gene Identification.

Most Hieracium subgenus Pilosella species are self-incompatible. Some undergo facultative apomixis where most seeds form asexually with a maternal genotype. Most embryo sacs develop by mitosis, without meiosis and seeds form without fertilization. Apomixis is controlled by dominant loci where recombination is suppressed. Loci deletion by γ-irradiation results in reversion to sexual reproduction. Targeted mutagenesis of genes at identified loci would facilitate causal gene identification. In this study, the efficacy of CRISPR/Cas9 editing was examined in apomictic Hieracium by targeting mutations in the endogenous PHYTOENE DESATURASE (PDS) gene using Agrobacterium-mediated leaf disk transformation. In three experiments, the expected albino dwarf-lethal phenotype, characteristic of PDS knockout, was evident in 11% of T0 plants, 31.4% were sectorial albino chimeras, and the remainder were green. The chimeric plants flowered. Germinated T1 seeds derived from apomictic reproduction in two chimeric plants were phenotyped and sequenced to identify PDS gene edits. Up to 86% of seeds produced albino seedlings with complete PDS knockout. This was attributed to continuing Cas9-mediated editing in chimeric plants during apomictic seed formation preventing Cas9 segregation from the PDS target. This successful demonstration of efficient CRISPR/Cas9 gene editing in apomictic Hieracium, enabled development of the discussed strategies for future identification of causal apomixis genes.

The Stopit! programme to reduce bullying and undermining behaviour in hospitals.

Purpose The impact of bullying and undermining behaviours on the National Health Service on costs, patient safety and retention of staff was well understood even before the Illing report, published in 2013, that reviewed the efficacy of training interventions designed to reduce bullying and harassment in the outputs. The purpose of this paper is to provide an example of a good programme well evaluated. Design/methodology/approach The methodology follows a broad realist approach, by specifying the underlying programme assumptions and intention of the designers. Three months after the event, Q-sort methodology was employed to group participants into one of three contexts - mechanism - output groups. Interviews were then undertaken with members of two of these groups, to evaluate how the programme had influenced each. Findings Q-sort identified a typology of three beneficiaries from the Stopit! workshops, characterised as professionals, colleagues and victims. Each group had acted upon different parts of the programme, depending chiefly upon their current and past experiences of bullying in hospitals. Research limitations/implications The paper demonstrates the effectiveness of using Q-sort method to identify relevant CMOs in a realist evaluation framework. Practical implications The paper considers the effectiveness of the programme to reduce bullying, rather than teach victims to cope, and how it may be strengthened based upon the research findings and Illing recommendations. Social implications Workplace bullying is invariably implicated in scandals concerning poor hospital practice, poor patient outcomes and staff illness. All too frequently, the sector responds by offering training in resilience, which though helpful, places the onus on the victim to cope rather than the employer to reduce or eliminate the practice. This paper documents and evaluates an attempt to change workplace practices to directly address bullying and undermining. Originality/value The paper describes a new programme broadly consistent with Illing report endorsements. Second, it illustrates a novel evaluation method that highlights rigorously the contexts, mechanisms and outcomes at the pilot stage of an intervention identifies contexts and mechanisms via factor analysis using Q-sort methodology.

Assembled genomic and tissue-specific transcriptomic data resources for two genetically distinct lines of Cowpea ( Vigna unguiculata (L.) Walp).

Cowpea ( Vigna unguiculata (L.) Walp) is an important legume crop for food security in areas of low-input and smallholder farming throughout Africa and Asia. Genetic improvements are required to increase yield and resilience to biotic and abiotic stress and to enhance cowpea crop performance. An integrated cowpea genomic and gene expression data resource has the potential to greatly accelerate breeding and the delivery of novel genetic traits for cowpea. Extensive genomic resources for cowpea have been absent from the public domain; however, a recent early release reference genome for IT97K-499-35 ( Vigna unguiculata v1.0, NSF, UCR, USAID, DOE-JGI, http://phytozome.jgi.doe.gov/) has now been established in a collaboration between the Joint Genome Institute (JGI) and University California (UC) Riverside. Here we release supporting genomic and transcriptomic data for two transformable cowpea varieties, IT97K-499-35 and IT86D-1010. The transcriptome resource includes six tissue-specific datasets for each variety, with particular emphasis on reproductive tissues that extend and support the V. unguiculata v1.0 reference. Annotations have been included in our resource to allow direct mapping to the v1.0 cowpea reference. The resource described here is supported by downloadable raw and assembled sequence data.

Consultants as victims of bullying and undermining: a survey of Royal College of Obstetricians and Gynaecologists consultant experiences.

OBJECTIVE: To explore incidents of bullying and undermining among obstetrics and gynaecology (O&G) consultants in the UK, to add another dimension to previous research and assist in providing a more holistic understanding of the problem in medicine. DESIGN: Questionnaire survey. SETTING: Royal College of Obstetricians and Gynaecologists (RCOG). PARTICIPANTS: O&G consultant members/fellows of the RCOG working in the UK. MAIN OUTCOME MEASURES: Measures included a typology of 4 bullying and undermining consequences from major to coping. RESULTS: There was a 28% (664) response rate of whom 44% (229) responded that they had been persistently bullied or undermined. Victims responded that bullying and undermining is carried out by those senior or at least close in the hierarchy. Of the 278 consultants who answered the question on 'frequency of occurrence', 50% stated that bullying and undermining occurs on half, or more, of all encounters with perpetrators and two-thirds reported that it had lasted more than 3 years. The reported impact on professional and personal life spans a wide spectrum from suicidal ideation, depression and sleep disturbance, and a loss of confidence. Over half reported problems that could compromise patient care. When victims were asked if the problem was being addressed, 73% of those that responded stated that it was not. CONCLUSIONS: Significant numbers of consultants in O&G in the UK are victims of bullying and undermining behaviour that puts their own health and patient care at risk. New interventions to tackle the problem, rather than its consequences, are required urgently, together with greater commitment to supporting such interventions.

An extended U2AF(65)-RNA-binding domain recognizes the 3' splice site signal.

How the essential pre-mRNA splicing factor U2AF(65) recognizes the polypyrimidine (Py) signals of the major class of 3' splice sites in human gene transcripts remains incompletely understood. We determined four structures of an extended U2AF(65)-RNA-binding domain bound to Py-tract oligonucleotides at resolutions between 2.0 and 1.5 Å. These structures together with RNA binding and splicing assays reveal unforeseen roles for U2AF(65) inter-domain residues in recognizing a contiguous, nine-nucleotide Py tract. The U2AF(65) linker residues between the dual RNA recognition motifs (RRMs) recognize the central nucleotide, whereas the N- and C-terminal RRM extensions recognize the 3' terminus and third nucleotide. Single-molecule FRET experiments suggest that conformational selection and induced fit of the U2AF(65) RRMs are complementary mechanisms for Py-tract association. Altogether, these results advance the mechanistic understanding of molecular recognition for a major class of splice site signals.

Specialty courts: Who's in and are they working?

The effectiveness of specialty courts has been well established in the literature; however, previous studies have not taken into account referral biases that may exist based on offenders' race, socioeconomic status (SES), attorney status, and so forth. The current study hypothesized that (a) Participants who are racially diverse, of lower SES, and represented by privately retained attorneys would be referred less frequently to specialty courts, and (b) Participants in specialty courts would evidence reductions in missed court appointments and failed urinary analyses (UAs) compared with peers not enrolled in specialty courts. Participants (N = 274) were probationers who were involved in 1 of 3 specialty court programs (i.e., drug, driving while intoxicated [DWI], or reentry courts) or a matched sample of probationers not in specialty court services. Results indicated that, in general and with few exceptions, specialty courts did not have differential referral rates based on offender demographics including race, ethnicity, SES, or attorney status (court appointed vs. privately retained). Results examining the effectiveness of the specialty courts were mixed. Participants in the in-prison treatment program reentry court missed a greater proportion of scheduled court meetings than did their matched sample counterparts; however, the other specialty court programs did not significantly differ from their matched-sample counterparts. Participants in the DWI court had a significantly smaller proportion of UA failures to total UAs than did their matched sample peers; however, the drug court and reentry court programs did not significantly differ from their matched sample counterparts. Implications, future directions, and limitations are discussed. (PsycINFO Database Record

A brief peripheral motion contrast threshold test predicts older drivers' hazardous behaviors in simulated driving.

Our research group has previously demonstrated that the peripheral motion contrast threshold (PMCT) test predicts older drivers' self-report accident risk, as well as simulated driving performance. However, the PMCT is too lengthy to be a part of a battery of tests to assess fitness to drive. Therefore, we have developed a new version of this test, which takes under two minutes to administer. We assessed the motion contrast thresholds of 24 younger drivers (19-32) and 25 older drivers (65-83) with both the PMCT-10min and the PMCT-2min test and investigated if thresholds were associated with measures of simulated driving performance. Younger participants had significantly lower motion contrast thresholds than older participants and there were no significant correlations between younger participants' thresholds and any measures of driving performance. The PMCT-10min and the PMCT-2min thresholds of older drivers' predicted simulated crash risk, as well as the minimum distance of approach to all hazards. This suggests that our tests of motion processing can help predict the risk of collision or near collision in older drivers. Thresholds were also correlated with the total lane deviation time, suggesting a deficiency in processing of peripheral flow and delayed detection of adjacent cars. The PMCT-2min is an improved version of a previously validated test, and it has the potential to help assess older drivers' fitness to drive.

Near peripheral motion contrast threshold predicts older drivers' simulator performance.

Our group has previously demonstrated that peripheral motion contrast threshold (PMCT) is significantly associated with self-reported accident risk of older drivers (questionnaire assessment), and with Useful Field of View(®) subtest 2 (UFOV2). It has not been shown, however, that PMCT is significantly associated with driving performance. Using the method of descending limits (spatial two-alternative forced choice) we assessed motion contrast thresholds of 28 young participants (25-45), and 21 older drivers (63-86) for 0.4 cycle/degree drifting Gabor stimuli at 15° eccentricity and examined whether it was related to performance on a simulated on-road test and to a measure of visual attention (UFOV(®) subtests 2 and 3). Peripheral motion contrast thresholds (PMCT) of younger participants were significantly lower than older participants. PMCT and UFOV2 significantly predicted driving examiners' scores of older drivers' simulator performance, as well as number of crashes. Within the older group, PMCT correlated significantly with UFOV2, UFOV3, and age. Within the younger group, PMCT was not significantly related to either UFOV(®) scores or age. Partial correlations showed that: substantial association between PMCT and UFOV2 was not age-related (within the older driver group); PMCT and UFOV2 tapped a common visual function; and PMCT assessed a component not captured by UFOV2. PMCT is potentially a useful assessment tool for predicting accident risk of older drivers, and for informing efforts to develop effective countermeasures to remediate this functional deficit as much as possible.

Genetic separation of autonomous endosperm formation (AutE) from the two other components of apomixis in Hieracium.

In apomictic Hieracium subgenus Pilosella species, embryo sacs develop in ovules without meiosis. Embryo and endosperm formation then occur without fertilization, producing seeds with a maternal genotype encased in a fruit (achene). Genetic analyses in H. praealtum indicate a dominant locus (LOA) controls meiotic avoidance, and another dominant locus (LOP) controls both fertilization-independent embryogenesis and endosperm formation. While cytologically examining developmental events in ovules of progeny from crosses between different wild-type and mutant Hieracium apomicts, and a sexual Hieracium species, we identified two plants, AutE196 and AutE24, which have lost the capacity for meiotic avoidance and fertilization-independent embryo formation. AutE196 and AutE24 exhibit autonomous endosperm formation and set parthenocarpic, seedless achenes at a penetrance of 18 %. Viable seed form after pollination. Cytological examination of 102 progeny from a backcross of AutE196 with sexual H. pilosella showed that autonomous endosperm formation is a heritable, dominant, qualitative trait, detected in 51 % of progeny. Variation in quantitative trait penetrance indicates other factors influence its expression. The correlation between autonomous endosperm development and mature parthenocarpic achene formation suggests the former is sufficient to trigger fruit maturation in Hieracium. The developmental component of autonomous endosperm formation is therefore genetically separable from those controlling meiotic avoidance and autonomous embryogenesis in Hieracium and has been denoted as AutE. We postulate that tight linkage of AutE and genes controlling autonomous embryogenesis at the LOP locus in H. praealtum may explain why inheritance of autonomous seed formation is typically observed as a single component.

Exploring the benefits of augmented reality documentation for maintenance and repair.

We explore the development of an experimental augmented reality application that provides benefits to professional mechanics performing maintenance and repair tasks in a field setting. We developed a prototype that supports military mechanics conducting routine maintenance tasks inside an armored vehicle turret, and evaluated it with a user study. Our prototype uses a tracked headworn display to augment a mechanic's natural view with text, labels, arrows, and animated sequences designed to facilitate task comprehension, localization, and execution. A within-subject controlled user study examined professional military mechanics using our system to complete 18 common tasks under field conditions. These tasks included installing and removing fasteners and indicator lights, and connecting cables, all within the cramped interior of an armored personnel carrier turret. An augmented reality condition was tested against two baseline conditions: the same headworn display providing untracked text and graphics and a fixed flat panel display representing an improved version of the laptop-based documentation currently employed in practice. The augmented reality condition allowed mechanics to locate tasks more quickly than when using either baseline, and in some instances, resulted in less overall head movement. A qualitative survey showed that mechanics found the augmented reality condition intuitive and satisfying for the tested sequence of tasks.