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Tropical islands hold great treasures of Earth's biodiversity, but these fragile ecosystems may be lost before their diversity is fully catalogued or the evolutionary processes that birthed it are understood. We ran comparative analyses on the ND2 and ND3 mitochondrial genes of the Sulawesi babbler Pellorneum celebense, an understorey bird endemic to Sulawesi and its continental islands, along with its morphology and song. Genetic, acoustic, and morphological data agree on multiple isolated populations, likely representing independently evolving lineages. The Sulawesi babbler shows signs of rapid speciation, with populations diverging between Central and Southeast Sulawesi, and even on land-bridge islands which were connected within the last few tens of thousands of years. The genetic divergence between Sulawesi babbler populations in this time has been around 33% of their divergence from sister species which have been isolated from Sulawesi for millions of years. This is likely facilitated by the Sulawesi babbler's understorey lifestyle, which inhibits gene flow and promotes speciation. Similar patterns of endemism are seen in Sulawesi's mammals and amphibians. This work highlights the undocumented biodiversity of a threatened hotspot, wrought by complex processes of speciation which interact with ecology and geology. Subspecific taxonomy has at times been controversial, but we argue that discrete populations such as these play a key role in evolution. Lying as they do at the heart of the biodiversity hotspot of Wallacea, these islands can reveal much about the evolution of biodiversity at all of its levels, from the gene to the ecosystem.

The size selectivity of four codends were compared in the Gulf of St. Lawrence, Canada, redfish fishery (Sebastes mentella), including the regulated diamond mesh codend with a mesh opening of 90 mm (T0) and three experimental codends of different mesh openings (90, 100, 110 mm) in which the netting is turned 90° to the direction of tow (T90). Results for the regulated codend showed that there was little size selection, catching greater than 97% of redfish over all of the length classes observed. Considering the fished population, the smallest T90 codend would catch 30% fewer redfish under the minimum landing size (MLS) of 22 cm compared with the T0 codend, but would also lose 16% of catch above 22 cm. The T90 codend with 100 mm mesh opening had the same size selectivity as the smallest T90 codend. The 110 mm T90 codend would catch 50% less redfish below MLS but lose 40% of redfish above MLS. Overall, results show that T90 codends improve size selectivity in which large proportions of undersized fish are successfully released.

The paralogues RrpA and RrpB, which are members of the MarR family of DNA binding proteins, are important for the survival of the global bacterial foodborne pathogen Campylobacter jejuni under redox stress. We report that RrpA is a positive regulator of mdaB, encoding a flavin-dependent quinone reductase that contributes to the protection from redox stress mediated by structurally diverse quinones, while RrpB negatively regulates the expression of cj1555c (renamed nfrA for NADPH-flavin reductase A), encoding a flavin reductase. NfrA reduces riboflavin at a greater rate than its derivatives, suggesting that exogenous free flavins are the natural substrate. MdaB and NfrA both prefer NADPH as an electron donor. Cysteine substitution and posttranslational modification analyses indicated that RrpA and RrpB employ a cysteine-based redox switch. Complete genome sequence analyses revealed that mdaB is frequently found in Campylobacter and related Helicobacter spp., while nfrA is predominant in C. jejuni strains. Quinones and flavins are redox cycling agents secreted by a wide range of cell types that can form damaging superoxide by one-electron reactions. We propose a model for stress adaptation where MdaB and NfrA facilitate a two-electron reduction mechanism to the less toxic hydroquinones, thus aiding survival and persistence of this major pathogen.\ud \ud \ud \ud IMPORTANCE Changes in cellular redox potential result in alteration in the oxidation state of intracellular metabolites and enzymes; consequently, cells make adjustments that favor growth and survival. The work we present here answers some of the many questions that have remained elusive over the years of investigation into the enigmatic microaerophile bacterium Campylobacter jejuni. We employed molecular approaches to understand the regulation mechanisms and functional analyses to reveal the roles of two novel quinone and flavin reductases; both serve as major pools of cellular redox-active molecules. This work extends our knowledge on bacterial redox sensing mechanisms and the significance of hemostasis.

In growing cattle, the phenotypic and genetic relationships between feeding behavior and both live animal ultrasound measures and subsequent carcass merit are generally poorly characterized. The objective of the current study was to quantify the phenotypic and genetic associations between a plethora of feeding behavior traits with both pre-slaughter ultrasound traits and post-slaughter carcass credentials in crossbred Bos taurus cattle. Carcass data were available on 3,146 young bulls, steers, and heifers, of which 2,795 and 2,445 also had records for pre-slaughter ultrasound muscle depth and intramuscular fat percentage, respectively; a total of 1,548 steers and heifers had information on all of the feeding behavior, ultrasound, and carcass traits. Young bulls were fed concentrates, while steers and heifers were fed a total mixed ration. Feeding behavior traits were defined based on individual feed events or meal events (i.e., individual feed events grouped into meals). Animal linear mixed models were used to estimate (co)variance components. Phenotypic correlations between feeding behavior and both ultrasound and carcass traits were generally weak and not different from zero, although there were phenotypic correlations of 0.40, 0.26, and 0.37 between carcass weight and feeding rate, energy intake per feed event, and energy intake per meal, respectively. Genetically, cattle that had heavier carcass weights, better carcass conformation, or a higher dressing percentage fed for a shorter time per day (genetic correlations [±SE] of −0.46 ± 0.12, −0.39 ± 0.11, and −0.50 ± 0.10, respectively). Genetic correlations of 0.43 ± 0.12 and 0.68 ± 0.13 were estimated between dressing difference (i.e., differential between live weight pre-slaughter and carcass weight) and energy intake per feed event and energy intake per meal, respectively. Neither intramuscular fat percentage measured on live animals nor carcass fat score (i.e., a measure of subcutaneous fat cover of the carcass) was genetically associated with any of the feeding behavior traits. The genetic associations between some feeding behavior traits and both ultrasound and carcass traits herein suggested that indirect responses in the feeding behavior of growing cattle from selection for improved carcass merit would be expected. Such changes in feeding behavior patterns in cattle may be reduced by measuring and including feeding behavior in a multiple-trait selection index alongside carcass traits.

To the best of our knowledge, the genetic variability in feeding behavior, as well as relationships with performance and feed efficiency, has not been investigated in a cattle population of greater than 1,500 animals. Our objective was to quantify the genetic parameters of several feeding behavior traits, and their genetic associations with both performance and feed efficiency traits, in crossbred growing cattle. Feed intake and live-weight data were available on 6,088 bulls, steers and heifers; of these, 4,672 cattle had backfat and muscle ultrasound data, and 1,548 steers and heifers had feeding behavior data. Genetic (co)variance parameters were estimated using animal linear mixed models; fixed effects included test group, heterosis, recombination loss, dam parity, age in months at the end of test, and the two-way interaction between age in months at the end of test and sex. Heritability was estimated to be 0.51 (0.097), 0.61 (0.100), 0.44 (0.093), 0.48 (0.094), and 0.47 (0.095) for feed events per day, feeding time per day, feeding rate, feed event duration, and energy intake per feed event, respectively. Coefficients of genetic variation ranged from 0.11 (feeding time per day) to 0.22 (feed event duration). Genetically heavier cattle with a higher energy intake per day, and faster growth rate, had a faster feeding rate and a greater energy intake per feed event. Genetic correlations between feeding behavior and feed efficiency were generally not different from zero, however, there was a genetic correlation of 0.36 (0.11) between feeding time per day and residual energy intake. Significant heritable and exploitable genetic variation exists in several feeding behavior traits in crossbred growing cattle which are also correlated with several performance traits. As some feeding behavior traits may be relatively less resource intensive to measure, they could be useful as predictor traits in beef cattle genetic evaluations.

The objectives of the present study were to estimate genetic parameters for several feeding behavior traits in growing cattle, as well as the genetic associations among and between feeding behavior and both performance and feed efficiency traits. An additional objective was to investigate the use of feeding behavior traits as predictors of genetic merit for feed intake. Feed intake and live-weight data on 6,088 growing cattle were used of which 4,672 had ultrasound data and 1,548 had feeding behavior data. Feeding behavior traits were defined based on individual feed events or meal events (where individual feed events were grouped into meals). Univariate and bivariate animal linear mixed models were used to estimate (co)variance components. Heritability estimates (± SE) for the feeding behavior traits ranged from 0.19 ± 0.08 for meals per day to 0.61 ± 0.10 for feeding time per day. The coefficient of genetic variation per trait varied from 5% for meals per day to 22% for the duration of each feed event. Genetically heavier cattle, those with a higher daily energy intake (MEI), or those that grew faster had a faster feeding rate, as well as a greater energy intake per feed event and per meal. Better daily feed efficiency (i.e., lower residual energy intake) was genetically associated with both a shorter feeding time per day and shorter meal time per day. In a validation population of 321 steers and heifers, the ability of estimated breeding values (EBV) for MEI to predict (adjusted) phenotypic MEI was demonstrated; EBVs for MEI were estimated using multi-trait models with different sets of predictor traits such as liveweight and/or feeding behaviors. The correlation (± SE) between phenotypic MEI and EBV for MEI marginally improved (P < 0.001) from 0.64 ± 0.03 to 0.68 ± 0.03 when feeding behavior phenotypes from the validation population were included in a genetic evaluation that already included phenotypic mid-test metabolic live-weight from the validation population. This is one of the largest studies demonstrating that significant exploitable genetic variation exists in the feeding behavior of young crossbred growing cattle; such feeding behavior traits are also genetically correlated with several performance and feed efficiency metrics. Nonetheless, there was only a marginal benefit to the inclusion of time-related feeding behavior phenotypes in a genetic evaluation for MEI to improve the precision of the EBVs for this trait.

Arcobacter anaerophilus was originally described as the first obligate anaerobe in this genus by Sasi Jyothsna et al. 2013. The complete genome sequence of the type strain of this species was determined and analysed. Genes characteristic for organisms capable of aerobic growth were identified, and the ability of the organism to grow under microaerobic and aerobic conditions was confirmed in two independent laboratories. The description of A. anaerophilus is thus emended and the wider ramifications of these findings are discussed.

European badgers (Meles meles) are group‐living mustelids implicated in the spread of bovine tuberculosis (TB) to cattle and act as a wildlife reservoir for the disease. In badgers, only a minority of individuals disperse from their natal social group. However, dispersal may be extremely important for the spread of TB, as dispersers could act as hubs for disease transmission. We monitored a population of 139 wild badgers over 7 years in a medium‐density population (1.8 individuals/km2). GPS tracking collars were applied to 80 different individuals. Of these, we identified 25 dispersers, 14 of which were wearing collars as they dispersed. This allowed us to record the process of dispersal in much greater detail than ever before. We show that dispersal is an extremely complex process, and measurements of straight‐line distance between old and new social groups can severely underestimate how far dispersers travel. Assumptions of straight‐line travel can also underestimate direct and indirect interactions and the potential for disease transmission. For example, one female disperser which eventually settled 1.5 km from her natal territory traveled 308 km and passed through 22 different territories during dispersal. Knowledge of badgers' ranging behavior during dispersal is crucial to understanding the dynamics of TB transmission, and for designing appropriate interventions, such as vaccination., Only a minority of badgers disperse, and the process has rarely been recorded in detail. However, dispersers could act as important hubs for tuberculosis transmission. Our GPS tracking data reveal the process of dispersal in much greater detail than ever before. They show that measurements of straight‐line distance between old and new social groups severely underestimate how far dispersers travel, the potential for intraspecific interactions and the potential for disease transmission. Understanding the process of dispersal is crucial to understanding disease dynamics and designing appropriate interventions.

Some definitions of feed efficiency such as residual energy intake (REI) and residual gain (RG) may not truly reflect production efficiency. The energy sinks used in the derivation of the traits include metabolic live-weight; producers finishing cattle for slaughter are, however, paid on the basis of carcass weight, as opposed to live-weight. The objective of the present study was to explore alternative definitions of REI and RG which are more reflective of production efficiency, and quantify their relationship with performance, ultrasound, and carcass traits across multiple breeds and sexes of cattle. Feed intake and live-weight records were available on 5,172 growing animals, 2,187 of which also had information relating to carcass traits; all animals were fed a concentrate-based diet representative of a feedlot diet. Animal linear mixed models were used to estimate (co)variance components. Heritability estimates for all derived REI traits varied from 0.36 (REICWF; REI using carcass weight and carcass fat as energy sinks) to 0.50 (traditional REI derived with the energy sinks of both live-weight and ADG). The heritability for the RG traits varied from 0.24 to 0.34. Phenotypic correlations among all definitions of the REI traits ranged from 0.90 (REI with REICWF) to 0.99 (traditional REI with REI using metabolic preslaughter live-weight and ADG). All were different (P < 0.001) from one suggesting reranking of animals when using different definitions of REI to identify efficient cattle. The derived RG traits were either weakly or not correlated (P > 0.05) with the ultrasound and carcass traits. Genetic correlations between the REI traits with carcass weight, dressing difference (i.e., live-weight immediately preslaughter minus carcass weight) and dressing percentage (i.e., carcass weight divided by live-weight immediately preslaughter) implies that selection on any of the REI traits will increase carcass weight, lower the dressing difference and increase dressing percentage. Selection on REICW (REI using carcass weight as an energy sink), as opposed to traditional REI, should increase the carcass weight 2.2 times slower but reduce the dressing difference 4.3 times faster. While traditionally defined REI is informative from a research perspective, the ability to convert energy into live-weight gain does not necessarily equate to carcass gain, and as such, traits such as REICW and REICWF provide a better description of production efficiency for feedlot cattle.

Road ecology has traditionally focused on the impact of in-situ and functional roads on wildlife. However, road construction also poses a major, yet understudied, threat and the implications for key aspects of animal behaviour are unknown. Badgers (Meles meles) have been implicated in the transmission of tuberculosis to cattle. There are concerns that environmental disturbances, including major road construction, can disrupt badger territoriality, promoting the spread of the disease to cattle. To address these knowledge gaps the ranging behaviour of a medium-density Irish badger population was monitored using GPS-tracking collars before, during, and after a major road realignment project that bisected the study area. We estimated badgers’ home range sizes, nightly distances travelled, and the distance and frequency of extra-territorial excursions during each phase of the study and quantified any changes to these parameters. We show that road construction had a very limited effect on ranging behaviour. A small increase in nightly distance during road construction did not translate into an increase in home range size, nor an increase in the distance or frequency of extra-territorial excursions during road construction. In addition, suitable mitigation measures to prevent badger deaths appeared to ensure that normal patterns of ranging behaviour continued once the new road was in place. We recommend that continuous badger-proof fencing be placed along the entire length of new major roads, in combination with appropriately sited underpasses. Our analysis supports the view that road construction did not cause badgers to change their ranging behaviour in ways likely to increase the spread of tuberculosis.

The Chromosomal Passenger Complex (CPC; consisting of Borealin, Survivin, INCENP and Aurora B kinase) and Shugoshin 1 (Sgo1) are key regulators of chromosome bi-orientation, a process essential for error-free chromosome segregation. Their functions rely on their ability to associate with centromeres. Two histone phosphorylations, histone H3 Thr3 (H3T3ph; directly recognised by Survivin) and histone H2A Thr120 (H2AT120ph; indirectly recognised via Sgo1), together with CPC’s intrinsic ability to bind nucleosome, facilitate CPC centromere recruitment. The molecular basis for CPC-Sgo1 binding and how their direct interaction influences CPC centromere localisation and function are lacking. Here, using an integrative structure-function approach, we show that the histone H3-like Sgo1 N-terminal tail interacts with Survivin acting as a hot-spot for CPC-Sgo1 assembly, while downstream Sgo1 residues, mainly with Borealin contributes for high affinity interaction. Disruption of the Sgo1 N-terminal tail-Survivin interaction abolished CPC-Sgo1 assembly in vitro and perturbed centromere localisation and function of CPC. Our findings provide evidence that CPC binding to Sgo1 and histone H3 N-terminal tail are mutually exclusive, suggesting that these interactions will likely take place in a spatially/temporally restricted manner and provide a rationale for the Sgo1-mediated ‘kinetochore proximal centromere’ pool of CPC.

Summary STimulator of INterferon Genes (STING) is an adaptor for cytoplasmic DNA sensing by cGAMP/cGAS that helps trigger innate immune responses (IIRs). Although STING is mostly localized in the ER, we find a separate inner nuclear membrane pool of STING that increases mobility and redistributes to the outer nuclear membrane upon IIR stimulation by transfected dsDNA or dsRNA mimic poly(I:C). Immunoprecipitation of STING from isolated nuclear envelopes coupled with mass spectrometry revealed a distinct nuclear envelope-STING proteome consisting of known nuclear membrane proteins and enriched in DNA- and RNA-binding proteins. Seventeen of these nuclear envelope STING partners are known to bind direct interactors of IRF3/7 transcription factors, and testing a subset of these revealed STING partners SYNCRIP, MEN1, DDX5, snRNP70, RPS27a, and AATF as novel modulators of dsDNA-triggered IIRs. Moreover, we find that SYNCRIP is a novel antagonist of the RNA virus, influenza A, potentially shedding light on reports of STING inhibition of RNA viruses., Graphical abstract, Highlights • A nuclear envelope pool of STING redistributes upon innate immune response activation • STING has a unique set of nuclear partners that contribute to innate immune responses • RNA- and DNA-binding STING partners may promote IRF3 transcription factor activation • STING nuclear partners may explain its protection against RNA viruses, Molecular physiology; Immunology; Virology; Cell biology

RNA–protein interactions are central to all gene expression processes and contribute to a variety of human diseases. Therapeutic approaches targeting RNA–protein interactions have shown promising effects on some diseases that are previously regarded as ‘incurable’. Here, we developed a fluorescent on-bead screening platform, RNA Pull-Down COnfocal NAnoscanning (RP-CONA), to identify RNA–protein interaction modulators in eukaryotic cell extracts. Using RP-CONA, we identified small molecules that disrupt the interaction between HuR, an inhibitor of brain-enriched miR-7 biogenesis, and the conserved terminal loop of pri-miR-7–1. Importantly, miR-7′s primary target is an mRNA of α-synuclein, which contributes to the aetiology of Parkinson’s disease. Our method identified a natural product quercetin as a molecule able to upregulate cellular miR-7 levels and downregulate the expression of α-synuclein. This opens up new therapeutic avenues towards treatment of Parkinson’s disease as well as provides a novel methodology to search for modulators of RNA–protein interaction.

Bacterial C-type haem-copper oxidases in the cbb3 family are widespread in microaerophiles, which exploit their high oxygen-binding affinity for growth in microoxic niches. In microaerophilic pathogens, C-type oxidases can be essential for infection, yet little is known about their biogenesis compared to model bacteria. Here, we have identified genes involved in cbb3-oxidase (Cco) assembly and activity in the Gram-negative pathogen Campylobacter jejuni, the commonest cause of human food-borne bacterial gastroenteritis. Several genes of unknown function downstream of the oxidase structural genes ccoNOQP were shown to be essential (cj1483c and cj1486c) or important (cj1484c and cj1485c) for Cco activity; Cj1483 is a CcoH homologue, but Cj1484 (designated CcoZ) has structural similarity to MSMEG_4692, involved in Qcr-oxidase supercomplex formation in Mycobacterium smegmatis. Blue-native polyacrylamide gel electrophoresis of detergent solubilised membranes revealed three major bands, one of which contained CcoZ along with Qcr and oxidase subunits. Deletion of putative copper trafficking genes ccoI (cj1155c) and ccoS (cj1154c) abolished Cco activity, which was partially restored by addition of copper during growth, while inactivation of cj0369c encoding a CcoG homologue led to a partial reduction in Cco activity. Deletion of an operon encoding PCuAC (Cj0909) and Sco (Cj0911) periplasmic copper chaperone homologues reduced Cco activity, which was partially restored in the cj0911 mutant by exogenous copper. Phenotypic analyses of gene deletions in the cj1161c–1166c cluster, encoding several genes involved in intracellular metal homeostasis, showed that inactivation of copA (cj1161c), or copZ (cj1162c) led to both elevated intracellular Cu and reduced Cco activity, effects exacerbated at high external Cu. Our work has therefore identified (i) additional Cco subunits, (ii) a previously uncharacterized set of genes linking copper trafficking and Cco activity, and (iii) connections with Cu homeostasis in this important pathogen.

Campylobacter jejuni is a microaerophilic zoonotic pathogen with an atypical respiratory Complex I that oxidizes a flavodoxin (FldA) instead of NADH. FldA is essential for viability and is reduced via pyruvate and 2-oxoglutarate oxidoreductases (POR/OOR). Here, we show that FldA can also be reduced by FqrB (Cj0559), an NADPH:FldA reductase. An fqrB deletion mutant was viable but displayed a significant growth defect. FqrB is related to flavoprotein reductases from Gram-positive bacteria that can reduce NrdI, a specialized flavodoxin that is needed for tyrosyl radical formation in NrdF, the beta subunit of class 1b-type (Mn) ribonucleotide reductase (RNR). However, C. jejuni possesses a single class Ia-type (Fe) RNR (NrdAB) that would be expected to be ferredoxin dependent. We show that CjFldA is an unusually high potential flavodoxin unrelated to NrdI, yet growth of the fqrB mutant, but not the wild-type or a complemented strain, was stimulated by low deoxyribonucleoside (dRNS) concentrations, suggesting FldA links FqrB and RNR activity. Using purified proteins, we confirmed the NrdB tyrosyl radical could be regenerated in an NADPH, FqrB, and FldA dependent manner, as evidenced by both optical and electron paramagnetic resonance (EPR) spectroscopy. Thus, FldA activates RNR in C. jejuni, partly explaining its essentiality.

Movement of the cell nucleus typically involves the cytoskeleton and either polymerization-based pushing forces or motor-based pulling forces. In the fission yeast Schizosaccharomyces pombe, nuclear movement and positioning are thought to depend on microtubule polymerization-based pushing forces. Here, we describe a novel, microtubule-independent, form of nuclear movement in fission yeast. Microtubule-independent nuclear movement is directed towards growing cell tips, and it is strongest when the nucleus is close to a growing cell tip, and weakest when the nucleus is far from that tip. Microtubule-independent nuclear movement requires actin cables but does not depend on actin polymerization-based pushing or myosin V-based pulling forces. The vesicle-associated membrane protein (VAMP)-associated proteins (VAPs) Scs2 and Scs22, which are critical for endoplasmic reticulum–plasma membrane contact sites in fission yeast, are also required for microtubule-independent nuclear movement. We also find that in cells in which microtubule-based pushing forces are present, disruption of actin cables leads to increased fluctuations in interphase nuclear positioning and subsequent altered septation. Our results suggest two non-exclusive mechanisms for microtubule-independent nuclear movement, which may help illuminate aspects of nuclear positioning in other cells., Summary: A new form of nuclear movement in fission yeast, termed microtubule-independent nuclear movement, that is dependent on actin cables and on proteins involved in ER–plasma membrane contacts is described.

Significance StatementGenetic exchange among bacteria shapes the microbial world. From the acquisition of antimicrobial resistance genes to fundamental questions about the nature of bacterial species, this powerful evolutionary force has preoccupied scientists for decades. However, the mixing of genes between species rests on a paradox. On one hand, promoting adaptation by conferring novel functionality, on the other potentially introducing disharmonious gene combinations (negative epistasis) that will be selected against. Taking an interdisciplinary approach to analyse natural populations of the enteric bacteriaCampylobacter, an ideal example of long-range admixture, we demonstrate that genes can independently transfer across species boundaries and rejoin in epistasis in a recipient genome. This challenges conventional ideas and highlights the possibility of single step evolution by saltation.AbstractRecombination of short DNA fragments via horizontal gene transfer (HGT) can both introduce beneficial alleles and create genomic disharmony through negative epistasis. For non-core (accessory) genes, the negative epistatic cost is likely to be minimal because the incoming genes have not co-evolved with the recipient genome. By contrast, for the core genome, interspecific recombination is expected to be rare because disruptive allelic replacement is likely to introduce negative epistasis. Why then is homologous recombination common in the core of bacterial genomes? To understand this enigma we take advantage of an exceptional model system, the common enteric pathogensCampylobacter jejuniandCampylobacter coli, that are known for very high magnitude interspecies gene flow in the core genome. As expected, HGT does indeed disrupt co-adapted allele pairings (negative epistasis). However, multiple HGT events enable recovery of the genome’s co-adaption between introgressing alleles, even in core metabolism genes (e.g., formate dehydrogenase). These findings demonstrate that, even for complex traits, genetic coalitions can be decoupled, transferred and independently reinstated in a new genetic background – facilitating transition between fitness peaks. In this example, the two-step recombinational process is associated withC. colithat are adapted to the agricultural niche.

Purpose We previously identified three microRNAs (miRNAs) with significantly increased expression in the serum of patients with age-related macular degeneration (AMD) compared with healthy controls. Our objective was to identify potential functional roles of these upregulated miRNAs (miR-19a, miR-126, and miR-410) in AMD, using computational tools for miRNAs prediction and identification, and to demonstrate the miRNAs target genes and signaling pathways. We also aim to demonstrate the pathologic role of isolated sera-derived exosomes from patients with AMD and controls using in vitro models. Methods miR-19a, miR-126, and miR-410 were investigated using bioinformatic approaches, including DIANA-mirPath and miR TarBase. Data on the resulting target genes and signaling pathways were incorporated with the differentially expressed miRNAs in AMD. Apoptosis markers, human apoptosis miRNAs polymerase chain reaction arrays and angiogenesis/vasculogenesis assays were performed by adding serum-isolated AMD patient or control patient derived exosomes into an in vitro human angiogenesis model and ARPE-19 cell lines. Results A number of pathways known to be involved in AMD development and progression were predicted, including the vascular endothelial growth factor signaling, apoptosis, and neurodegenerative pathways. The study also provides supporting evidence for the involvement of serum-isolated AMD-derived exosomes in the pathology of AMD, via apoptosis and/or angiogenesis. Conclusions miR-19a, miR-126, miR-410 and their target genes had a significant correlation with AMD pathogenesis. As such, they could be potential new targets as predictive biomarkers or therapies for patients with AMD. Translational relevance The functional analysis and the pathologic role of altered miRNA expression in AMD may be applicable in developing new therapies for AMD through the disruption of individual or multiple pathophysiologic pathways.

Su et al. show that SUMOylation stabilizes bioriented kinetochore–microtubule attachments in mitosis to allow timely anaphase onset. They identify the shugoshin pericentromeric adaptor protein and the chromosome passenger complex as SUMOylation targets, which lead to dampening of error correction pathways., During mitosis, sister chromatids attach to microtubules from opposite poles, called biorientation. Sister chromatid cohesion resists microtubule forces, generating tension, which provides the signal that biorientation has occurred. How tension silences the surveillance pathways that prevent cell cycle progression and correct erroneous kinetochore–microtubule attachments remains unclear. Here we show that SUMOylation dampens error correction to allow stable sister kinetochore biorientation and timely anaphase onset. The Siz1/Siz2 SUMO ligases modify the pericentromere-localized shugoshin (Sgo1) protein before its tension-dependent release from chromatin. Sgo1 SUMOylation reduces its binding to protein phosphatase 2A (PP2A), and weakening of this interaction is important for stable biorientation. Unstable biorientation in SUMO-deficient cells is associated with persistence of the chromosome passenger complex (CPC) at centromeres, and SUMOylation of CPC subunit Bir1 also contributes to timely anaphase onset. We propose that SUMOylation acts in a combinatorial manner to facilitate dismantling of the error correction machinery within pericentromeres and thereby sharpen the metaphase–anaphase transition.

Nutrition knowledge is a key factor for consideration when evaluating the dietary intake of athletes. Positive associations have been established between higher nutrition knowledge and improved quality of dietary intake. Given the negative impact poor nutrition can have on performance and training adaptation, further investigation into athletes’ nutrition knowledge is warranted. Gaelic football is a field-based invasion team sport and players represent a unique sporting population due to their quasi-professional status. Inadequacies in players dietary intake have been observed, however no assessment of nutrition knowledge has been reported. This study examined players knowledge and compared results by playing level, education level, and history of nutrition education. An online survey was disseminated to a sample of male Gaelic football players (n = 152, mean age = 24.5 ± 5.9). This included 68 club (sub-elite) and 84 inter-county players (elite). Total score was 44.3 ± 12.7%, classified as “poor” and lower than previous findings from similar team sports. Significance was set at p

Despite advances in herd management, tuberculosis (TB) continues to affect ~0. 5% of Ireland's national cattle herd annually. It is clear that any “final” eradication of TB in cattle will need to address all TB maintenance hosts in the same environment. In Ireland and the UK, European Badgers (Meles meles) are a known TB maintenance host, while deer are recognised as spillover hosts. However, deer have been identified as maintenance hosts in other countries and Sika deer, specifically, have been identified with TB in Ireland. We examined the power of cattle, badger and Sika deer densities (at the county level) to predict cattle TB-breakdowns in Ireland, at both the herd and the individual level, using data collected between 2000 and 2018. Our hypothesis was that any positive correlations between deer density and cattle TB-breakdowns would implicate deer as TB maintenance hosts. Using linear multiple regressions, we found positive correlations between deer density and cattle TB-breakdowns at both the herd and individual levels. Since Sika deer in County Wicklow are known to have TB, we ran further regressions against subsets of data which excluded individual Irish counties. Analyses excluding Wicklow data showed much weaker correlations between Sika deer density and cattle TB-breakdowns at both the herd and individual levels, suggesting that these correlations are strongest in County Wicklow. A similar effect for badger density was seen in County Leitrim. While locally high densities of Sika deer persist in Irish counties, we believe they should be considered an integral part of any TB-control programme for those areas.

Movement of the cell nucleus typically involves the cytoskeleton and either polymerization-based pushing forces or motor-based pulling forces. In fission yeastSchizosaccharomyces pombe, nuclear movement and positioning are thought to depend on microtubule polymerization-based pushing forces. Here we describe a novel, microtubule-independent, form of nuclear movement in fission yeast. Microtubule-independent nuclear movement is directed towards growing cell tips, and it is strongest when the nucleus is close to a growing cell tip, and weakest when the nucleus is far from that tip. Microtubule-independent nuclear movement requires actin cables but does not depend on actin polymerization-based pushing or myosin V-based pulling forces. Vesicle-associated membrane protein (VAMP)-associated proteins (VAPs) Scs2 and Scs22, which are critical for endoplasmic reticulum-plasma membrane contact sites in fission yeast, are also required for microtubule-independent nuclear movement. We also find that in cells in which microtubule-based pushing forces are present, disruption of actin cables leads to increased fluctuations in interphase nuclear positioning and subsequent altered septation. Our results suggest two non-exclusive mechanisms for microtubule-independent nuclear movement, which may help illuminate aspects of nuclear positioning in other cells.

The objective was to determine the factors associated with meat tenderness, juiciness, flavour and chewiness in 4791 growing crossbred cattle. Meat quality of bulls was inferior to that of both steers and heifers with little difference between the latter two genders. Angus, Hereford and Belgian Blues had the most tender meat with the Simmental being the toughest albeit the difference was, on average, only 5%. Moderate to strong correlations (r ≥ |0.43|) existed among tenderness, juiciness and flavour although some of the correlations differed by animal gender. Correlations between chewiness and tenderness in the different genders varied from −0.81 to −0.74 while the correlations between chewiness and the other sensory traits varied from −0.54 to −0.09. The (partial) correlations between each of the four sensory metrics and all of carcass weight, carcass conformation and carcass fat score were ≤|0.09| with most not being different from zero. Correlations between the sensory traits with growth rate, muscle depth, feed intake and efficiency were all ≤|0.08| and mostly not different from zero.

The purpose of this study was to define an extensive suite of feeding behavior traits in growing crossbred cattle and to investigate their phenotypic inter-relationships as well as relationships with other performance and efficiency traits. Time-series feeding behavior data, as well as feed intake and liveweight records, were available for 624 growing crossbred cattle, of which 445 were steers and 179 were heifers. Feeding behavior repeatability estimates were calculated using linear mixed models. Additionally, partial Spearman correlations were estimated among 14 feeding behavior traits, as well as between feeding behavior with both performance and feed efficiency traits, using residuals retained from linear mixed models. The marginal contribution of several feeding behavior traits to the variability in metabolizable energy intake (MEI) was also determined. Repeatability estimates of 0.57, 0.36, and 0.48 were calculated for the number of feed events per day, the total time spent feeding per day, and the feeding rate, respectively. Cattle that ate more frequently each day, ate at a faster rate and consumed less energy in each visit to the feed bunk. More efficient cattle fed less often per day and fed for a shorter duration per day; they also had a slower feeding rate and fed for longer in each visit to the feed bunk. Moreover, heavier cattle fed for a longer duration per day had a faster feeding rate, but fed less often per day; heavier animals also fed first in the pen after the fresh feed was offered. The number of feed events per day and feeding time per day together explained an additional 13.4 percentage points of the variability in MEI above that already explained by all of growth rate, liveweight, and backfat depth. The results from the present study suggest that several repeatable time-series-related feeding behavior traits, that are less resource intensive to measure, may have a role as useful predictor traits of important but relatively difficult to record traits, such as feed intake and efficiency.

Few studies have attempted to quantify the association between a terminal total merit index with phenotypic feed and production efficiency in beef cattle, particularly when feed efficiency is itself explicitly absent as a goal trait in the index. The objective of the present study was to quantify the differences in phenotypic performance for feed intake, feed efficiency, and carcass traits of crossbred bulls, steers, and heifers differing in a terminal total merit index. A validation population of 614 bulls, steers, and heifers that were evaluated for feed intake and efficiency in the same feedlot and subsequently slaughtered at the end of their test period was constructed. The Irish national genetic evaluations for a terminal index of calving performance, docility, feed intake, and carcass traits were undertaken with the phenotypic records of animals present in the validation population masked. The validation population animals were subsequently stratified into four groups, within sex, according to their terminal index value. Mixed models were used to quantify the association between terminal genetic merit and phenotypic performance; whether the associations differed by sex were also investigated. The regression coefficient of phenotypic feed intake, carcass weight, carcass conformation, or carcass fat on its respective estimated breeding values was 0.86 kg dry matter 0.91 kg, 1.01 units, and 1.29 units, respectively, which are close to the expectation of one. On average, cattle in the very high terminal index stratum had a 0.63 kg DM/d lower feed intake, a 25.05 kg heavier carcass, a 1.82 unit better carcass conformation (scale 1 to 15), and a 1.24 unit less carcass fat score (scale 1 to 15), relative to cattle in the very low terminal index stratum. Cattle of superior total genetic merit were also more feed efficient (i.e., had a lower energy conversion ratio, lower residual feed intake, and greater residual gain), had a greater proportion of their live-weight as carcass weight (i.e., better dressing percentage) and were slaughtered at a younger age relative to their inferior total genetic merit counterparts. This study provides validation of an all-encompassing total merit index and demonstrates the benefits of selection on a total merit index for feed and production efficiency, which should impart confidence among stakeholders in the contribution of genetic selection to simultaneous improvements in individual animal performance and efficiency.

Competition takes place not only between species but also within them. Intersexual competition for resources may increase sexual dimorphism in body size to minimise ecological niche overlap. Change...

The bacterial flagellum is a remarkable molecular motor, whose primary function in bacteria is to facilitate motility through the rotation of a filament protruding from the bacterial cell. A cap complex, consisting of an oligomer of the protein FliD, is localized at the tip of the flagellum, and is essential for filament assembly, as well as adherence to surfaces in some bacteria. However, the structure of the intact cap complex, and the molecular basis for its interaction with the filament, remains elusive. Here we report the cryo-EM structure of the Campylobacter jejuni cap complex, which reveals that FliD is pentameric, with the N-terminal region of the protomer forming an extensive set of contacts across several subunits, that contribute to FliD oligomerization. We also demonstrate that the native C. jejuni flagellum filament is 11-stranded, contrary to a previously published cryo-EM structure, and propose a molecular model for the filament-cap interaction., FliD forms a cap complex at the tip of bacterial flagella and is essential for flagellum filament assembly. Here, the authors present the cryo-EM structure of the Campylobacter jejuni cap complex, revealing a pentameric assembly of FliD and further show that the C. jejuni flagellum filament is 11-stranded.

European badgers (Meles meles) in medium and high density populations show strong territorial behaviour. Territories in these populations are contiguous, well-marked and often unchanging over many years. However, badgers do not always stay within their territorial boundaries. In our medium-density population, most individual badgers made extra-territorial excursions (ETEs) throughout the year. ETEs were most frequent between April and September and least frequent in December and January (the period of winter lethargy). Male badgers made longer and more frequent ETEs than females (especially between January and March, and in autumn). Breeding females made longer and more frequent ETEs than non-breeding females in November. While these peaks correspond with the main mating seasons, mating activity does not explain ETEs throughout the year. The shorter, but more frequent, ETEs in summer months may serve a monitoring purpose, rather than simply providing additional mating opportunities with badgers from outside the ‘home’ social group. We found that young badgers did not make regular ETEs until the summer of their second year. If badgers could be vaccinated as cubs, this would reduce any potential risk of TB spread during ETEs.

The 3D architecture of the genome governs its maintenance, expression and transmission. The conserved ring-shaped cohesin complex organises the genome by topologically linking distant loci on either a single DNA molecule or, after DNA replication, on separate sister chromatids to provide the cohesion that resists the pulling forces of spindle microtubules during mitosis1,2. Cohesin is highly enriched in specialized chromosomal domains surrounding centromeres, called pericentromeres3-7. However, the structural organisation of pericentromeres and implications for chromosome segregation are unknown. Here we report the 3D structure of budding yeast pericentromeres and establish the relationship between genome organisation and function. We find that convergent genes mark pericentromere borders and, together with core centromeres, define their structure and function by positioning cohesin. Centromeres load cohesin and convergent genes at pericentromere borders trap it. Each side of the pericentromere is organised into a looped conformation, with border convergent genes at the base. Microtubule attachment extends a single pericentromere loop, size-limited by convergent genes at its borders. Re-orienting genes at borders into a tandem configuration repositions cohesin, enlarges the pericentromere and impairs chromosome biorientation in mitosis. Thus, the linear arrangement of transcriptional units together with targeted cohesin loading at centromeres shapes pericentromeres into a structure competent for chromosome segregation during mitosis. Our results reveal the architecture of the chromosomal region within which kinetochores are embedded and the re-structuring caused by microtubule attachment. Furthermore, we establish a direct, causal relationship between 3D genome organization of a specific chromosomal domain and cellular function.

The Tripartite Tricarboxylate Transporter (TTT) family is a poorly characterised group of prokaryotic secondary solute transport systems, which employ a periplasmic substrate binding-protein (SBP) for initial ligand recognition. The substrates of only a small number of TTT systems are known and very few SBP structures have been solved, so the mechanisms of SBP-ligand interactions in this family are not well understood. The SBP RPA4515 (AdpC) from Rhodopseudomonas palustris was found by differential scanning fluorescence and isothermal titration calorimetry to bind aliphatic dicarboxylates of a chain length of six to nine carbons, with KD values in the μM range. The highest affinity was found for the C6-dicarboxylate adipate (1,6-hexanedioate). Crystal structures of AdpC with either adipate or 2-oxoadipate bound revealed a lack of positively charged amino-acids in the binding pocket and showed that water molecules are involved in bridging hydrogen bonds to the substrate, a conserved feature in the TTT SBP family that is distinct from other types of SBP. In AdpC, both of the ligand carboxylate groups and a linear chain conformation are needed for coordination in the binding pocket. RT-PCR showed that adpC expression is upregulated by low environmental adipate concentrations, suggesting adipate is a physiologically relevant substrate but as adpC is not genetically linked to any TTT membrane transport genes, the role of AdpC may be in signalling rather than transport. Our data expands the known ligands for TTT systems and identifies a novel high-affinity binding-protein for adipate, an important industrial chemical intermediate and food additive. This article is protected by copyright. All rights reserved.

A gene expression-based siRNA screen was used to evaluate functional similarity between genetic perturbations to identify functionally similar proteins. A siRNA library (siGenome library, Dharmacon) consisting of multiple siRNAs per gene that have been pooled in to one well per gene was arrayed in a 384-well format and used to individually target 14,335 proteins for depletion in HCT116 colon cancer cells. For each protein depletion, the gene expression of eight genes was quantified using the multiplexed Affymetrix Quantigene 2.0 assay in technical triplicate. As a proof of concept, six genes (BNIP3, NDRG1, ALDOC, LOXL2, ACSL5, BNIP3L) whose expression pattern reliably reflect the disruption of the molecular scaffold KSR1 were measured upon each protein depletion. The remaining two genes (PPIB and HPRT) are housekeeping genes used for normalization. The gene expression signatures from this screen can be used to estimate the functional similarity between any two proteins and successfully identified functional relationships for specific proteins such as KSR1 and more generalized processes, such as autophagy., Measurement(s)gene expression • KSR1 pathway regulationTechnology Type(s)Microarray • RNA interferenceFactor Type(s)target RNASample Characteristic - OrganismHomo sapiens Machine-accessible metadata file describing the reported data: 10.6084/m9.figshare.11418018

Significance Human activities have had a profound effect on the Earth’s ecosystems and biodiversity. This is evident among livestock species, such as cattle, that now constitute more biomass than all wild mammals combined. For bacteria living within these animals, this massive host expansion represents an opportunity to proliferate and spread globally through trade networks. This carries significant risks as pathogenic species can potentially spread to humans, but little is known about how these zoonotic bacteria evolve to exploit the changing host niche space. Here, we explain the nature and timescale of cattle adaptation in the world’s most common enteric bacterial pathogen (Campylobacter jejuni), highlighting potential risks of intensive livestock production in pathogen emergence., Modern agriculture has dramatically changed the distribution of animal species on Earth. Changes to host ecology have a major impact on the microbiota, potentially increasing the risk of zoonotic pathogens being transmitted to humans, but the impact of intensive livestock production on host-associated bacteria has rarely been studied. Here, we use large isolate collections and comparative genomics techniques, linked to phenotype studies, to understand the timescale and genomic adaptations associated with the proliferation of the most common food-born bacterial pathogen (Campylobacter jejuni) in the most prolific agricultural mammal (cattle). Our findings reveal the emergence of cattle specialist C. jejuni lineages from a background of host generalist strains that coincided with the dramatic rise in cattle numbers in the 20th century. Cattle adaptation was associated with horizontal gene transfer and significant gene gain and loss. This may be related to differences in host diet, anatomy, and physiology, leading to the proliferation of globally disseminated cattle specialists of major public health importance. This work highlights how genomic plasticity can allow important zoonotic pathogens to exploit altered niches in the face of anthropogenic change and provides information for mitigating some of the risks posed by modern agricultural systems.

Improvements in nutrition knowledge have been associated with increased carbohydrate consumption and greater adherence to dietary recommendations among female athletes. In order to assess whether nutrition knowledge interventions in female Gaelic games players may be beneficial, it is necessary to first of all investigate current levels of nutrition knowledge in this population. Given that many demographic characteristics have been shown to influence nutrition knowledge, it is also important for these to be investigated. The aims of this study were to evaluate the nutrition knowledge of female Gaelic games players, compare knowledge by players&rsquo, characteristics and identify players&rsquo, preferences for information and support. A validated 35-item questionnaire was completed by 328 female Gaelic games players (Age: 23.7 &plusmn, 5.0 years). Players&rsquo, mean nutrition knowledge score was 46.0% &plusmn, 11.8% and classified as &ldquo, poor&rdquo, Elite players scored greater (+4.5%&ndash, 5.9%, p &lt, 0.05) than subelite players. Players with higher levels of general education, history of formal nutrition education and previous advice from a nutritionist also presented greater nutrition knowledge (+3.7%&ndash, 7.5%, p &lt, 0.05). Future education interventions with female Gaelic games players may lead to beneficial changes in dietary behaviour and would likely benefit from stratifying content based on athletes&rsquo, demographic characteristics, given the differences observed.

This study investigated acute changes in markers of fatigue and performance attenuation during and following a competitive senior club-level Gaelic football match. Forty-one players were tested immediately pre-match, at half-time, full-time, 24 h post-match and 48 h post-match. Creatine kinase, drop jump height and contact-time, reactive strength index, countermovement jump height and perceptual responses were assessed at the aforementioned time-points. 18 Hz global positioning system devices were used to record players in-game workload measures. Compared to pre-match, perceptual responses (&minus, 27.6%) and countermovement jump height (&minus, 3.9%) were significantly reduced at full-time (p &lt, 0.05). Drop jump height (&minus, 8.8%), perceptual responses (&minus, 27.6%), reactive strength index (&minus, 15.6%) and countermovement jump height (&minus, 8.6%) were significantly lower 24 h post-match (p &lt, 0.05). Pre-match creatine kinase was significantly increased (+16.2% to +159.9%) when compared to all other time-points (p &lt, 0.05). Total distance, total accelerations, total sprints, sprint distance and average heart rate were all correlated to changes in perceptual responses (r = 0.34 to 0.56, p &lt, 0.05). Additionally, maximum speed achieved (r = 0.34) and sprint distance (r = 0.31) were significantly related to countermovement jump changes (p &lt, 0.05), while impacts (r = 0.36) were correlated to creatine kinase increases (p &lt, 0.05). These results demonstrate that Gaelic football match-play elicits substantial neuromuscular, biochemical and perceptual disturbances.

RationaleTransforming growth factor-beta (TGFβ) is tightly regulated at multiple levels, with regulation at the receptor level now recognized as a key determinant of the cellular response to this pleiotropic cytokine. TGFβ promotes saphenous vein graft neointima formation after coronary artery bypass graft (CABG) surgery, inducing smooth muscle cell (SMC) hyperplasia and fibrosis by signaling via activin receptor-like kinase 5(ALK5). However, the role of the alternate TGFβ receptor ALK1 remains completely unknown.ObjectiveTo define the receptor pathways activated by TGFβ in SMCs and their mechanistic importance during CABG neointima formation.Methods and resultsRadioligand co-IP assays revealed direct interactions between TGFβ, ALK5 and ALK1 in primary saphenous vein graft SMC (HSVSMC) from patients undergoing CABG. Knockdown and pharmacological inhibition of ALK5 or ALK1 in HSVSMC significantly attenuated TGFβ-induced phosphorylation of receptor-regulated (R)-Smads 2/3 and 1/5, respectively. Microarray profiling followed by qRT-PCR validation showed that TGFβ induced distinct transcriptional networks downstream of ALK5 or ALK1, associated with HSVSMC contractility and migration, respectively and confirmed using migration assays as well as qRT-PCR and western blot assays of contractile SMC markers. scRNAseq analysis of TGFβ-treated HSVSMC identified distinct subgroups of cells showing ALK5 or ALK1 transcriptional responses, while RNA velocity analyses indicated divergence in differentiation towards ALK5 or ALK1-dominant lineages. ALK1, ALK5 and their downstream effectors pSmad1/5 and pSmad2/3 were localized to αSMA+ neointimal SMCs in remodelled mouse vein grafts. Pharmacological inhibition or genetic ablation of Smad1/5 substantially reducing neointima formation following acute vascular injury. Notably, expression and activation of ALK1, ALK5 and their respective downstream R-Smads was already evident in hyperplastic saphenous veins prior to grafting.ConclusionsWhilst canonical TGFβ signaling via ALK5 promotes a contractile HSVSMC phenotype, transactivation of ALK1 by TGFβ induces neointima formation by driving cell migration. Restoring the balance between ALK1 and ALK5 in HSVSMC may represent a novel therapeutic strategy for vein graft failure.

Campylobacter jejuni outer membrane vesicles (OMVs) contain numerous virulence-associated proteins including the cytolethal distending toxin and three serine proteases. As C. jejuni lacks the classical virulence-associated secretion systems of other enteric pathogens that deliver effectors directly into target cells, OMVs may have a particularly important role in virulence. C. jejuni OMV production is stimulated by the presence of physiological concentrations of the bile salt sodium taurocholate (ST) through an unknown mechanism. The maintenance of lipid asymmetry (MLA) pathway has been implicated in a novel mechanism for OMV biogenesis, open to regulation by host signals. In this study we investigated the role of the MLA pathway in C. jejuni OMV biogenesis with ST as a potential regulator. OMV production was quantified by analyzing protein and lipid concentrations of OMV preparations and OMV particle counts produced by nanoparticle tracking analysis. Mutation of mlaA which encodes the outer membrane component of the MLA pathway significantly increased OMV production compared to the wild-type strain. Detergent sensitivity and membrane permeability assays confirmed the increased OMV production was not due to changes in membrane stability. The presence of 0.2% (w/v) ST increased wild-type OMV production and reduced OMV size, but did not further stimulate mlaA mutant OMV production or significantly alter mlaA mutant OMV size. qRT-PCR analysis demonstrated that the presence of ST decreased expression of both mlaA and mlaC in C. jejuni wild-type strains 11168 and 488. Collectively the data in this study suggests C. jejuni can regulate OMV production in response to host gut signals through changes in expression of the MLA pathway. As the gut bile composition is dependent on both diet and the microbiota, this study highlights the potential importance of diet and lifestyle factors on the varying disease presentations associated with gut pathogen infection.

Field-based team sports present large energetic demands given their intermittent high-intensity nature. Current evidence suggests that the dietary intake of female athletes may be insufficient to meet such demands, resulting in negative consequences for athletic performance and health. The primary aim of this review was to therefore assess the adequacy of dietary intake of female field-based team sport athletes when compared to dietary recommendations. A systematic search of databases, including PubMed, Web of Science, SPORTDiscus, and OpenGrey, was performed from the earliest record available until July 2020, obtaining an initial total of 2588 articles. To be included within the final review, articles were required to provide a quantitative assessment of baseline dietary intake specific to the target population. A total of 20 studies (n = 462) met the full eligibility criteria. A majority reported that the dietary intake of female field-based team sport athletes was insufficient in overall energy (2064 ± 309 kcal·day−1), carbohydrate (4.3 ± 1.2 g·kg·day−1), and iron intake (13.6 ± 6.2 mg·day−1) when compared to recommendations. Future research is required to establish why female team sport athletes consistently demonstrate deficient dietary practices, and to explore the potential negative consequences of this.

A series of laboratory-scale combustion tests were conducted under well-controlled conditions to measure the release of 90 Sr and 137 Cs nuclides to the atmosphere (air) from combustion of vegetation and organic soil samples contaminated with radioactivity. These vegetation and soil samples were collected from a controlled contaminated forest area within the Canadian Nuclear Laboratories – Chalk River site. The combustion products including ash and smoke particulates, along with gaseous emissions, were collected and then analyzed for 137 Cs and 90 Sr concentrations by radiometric techniques. The experimental results reveal that the releases of 90 Sr to the atmosphere (air) from combustion of vegetation are very low with most of the 90 Sr activity remaining in ash residues, even at a temperature of 800 °C. The detailed combustion experiments with surface litter and twigs, alder twigs, alder leaves, and organic soil indicate that 0.5 ± 0.1%, 0.3 ± 0.1%, 0.9 ± 0.1%, and 0.3 ± 0.1% of 90 Sr is released to the atmosphere (air), respectively. On the other hand, the releases of 137 Cs are found to be highly dependent on the combustion temperature as well as the nature of vegetation. The releases of 137 Cs obtained at 800 °C are 45 ± 7%, 77 ± 9%, 92 ± 5%, and 2.4 ± 0.5% for surface litter and twigs, alder twigs, alder leaves, and organic soil, respectively. The mechanism associated with the high release of 137 Cs at a high temperature of 800 °C was explored.

Identification and characterization of survival pathways active in tumor cells but absent in normal tissues provide opportunities to develop effective anticancer therapies with reduced toxicity to the patient. We show here that, like kinase suppressor of Ras 1 (KSR1), EPH (erythropoietin-producing hepatocellular carcinoma) receptor B4 (EPHB4) is aberrantly overexpressed in human colon tumor cell lines and selectively required for their survival. KSR1 and EPHB4 support tumor cell survival by promoting the expression of downstream targets, Myc and the transcriptional coactivator peroxisome proliferator-activated receptor gamma coactivator 1β (PGC1β). While KSR1 promotes the aberrant expression of Myc and the PGC1β protein via a posttranscriptional mechanism, EPHB4 has a greater effect on Myc and PGC1β expression via its ability to elevate mRNA levels. Subsequent analysis of the posttranscriptional regulation demonstrated that KSR1 promotes the translation of Myc protein. These findings reveal novel KSR1- and EPHB4-dependent signaling pathways supporting the survival of colorectal cancer cells through regulation of Myc and PGC1β, suggesting that inhibition of KSR1 or EPHB4 effectors may lead to selective toxicity in colorectal tumors.

Summary Whether gene repositioning to the nuclear periphery during differentiation adds another layer of regulation to gene expression remains controversial. Here, we resolve this by manipulating gene positions through targeting the nuclear envelope transmembrane proteins (NETs) that direct their normal repositioning during myogenesis. Combining transcriptomics with high-resolution DamID mapping of nuclear envelope-genome contacts, we show that three muscle-specific NETs, NET39, Tmem38A, and WFS1, direct specific myogenic genes to the nuclear periphery to facilitate their repression. Retargeting a NET39 fragment to nucleoli correspondingly repositioned a target gene, indicating a direct tethering mechanism. Being able to manipulate gene position independently of other changes in differentiation revealed that repositioning contributes ⅓ to ⅔ of a gene’s normal repression in myogenesis. Together, these NETs affect 37% of all genes changing expression during myogenesis, and their combined knockdown almost completely blocks myotube formation. This unequivocally demonstrates that NET-directed gene repositioning is critical for developmental gene regulation., Graphical Abstract, Highlights • Tissue-specific NETs direct repositioning of critical muscle genes during myogenesis • Expression changes for NET-repositioned genes depend on cell differentiation state • Isolating position from differentiation reveals its contribution to gene expression • Three NETs together affect 37% of all genes normally changing in myogenesis, Muscle-specific nuclear envelope transmembrane proteins (NETs) optimize myogenic gene expression by physically recruiting genes to the periphery and enhancing their repression. Specifically manipulating the position of endogenous genes in myoblasts and myotubes indicates that peripheral localization enhances repression, but only in context of other changes in differentiation.

Kinetochores direct chromosome segregation in mitosis and meiosis. Faithful gamete formation through meiosis requires that kinetochores take on new functions that impact homolog pairing, recombination and the orientation of kinetochore attachment to microtubules in meiosis I. Using an unbiased proteomics pipeline, we determined the composition of centromeric chromatin and kinetochores at distinct cell-cycle stages, revealing extensive reorganisation of kinetochores during meiosis. The data uncover a network of meiotic chromosome axis and recombination proteins that replace the microtubule-binding outer kinetochore sub-complexes during meiotic prophase. We show that this kinetochore remodelling in meiosis requires the Ctf19cCCAN inner kinetochore complex. Through functional analyses, we identify a Ctf19cCCAN-dependent kinetochore assembly pathway that is dispensable for mitotic growth, but becomes critical upon meiotic entry. Therefore, extensive kinetochore remodelling and a distinct assembly pathway direct the specialization of meiotic kinetochores for successful gametogenesis.

Campylobacter jejuni is a zoonotic Epsilonproteobacterium that grows in the gastrointestinal tract of birds and mammals, and is the most frequent cause of food-borne bacterial gastroenteritis worldwide. As an oxygen-sensitive microaerophile, C. jejuni has to survive high environmental oxygen tensions, adapt to oxygen limitation in the host intestine and resist host oxidative attack. Despite its small genome size, C. jejuni is a versatile and metabolically active pathogen, with a complex and highly branched set of respiratory chains allowing the use of a wide range of electron donors and alternative electron acceptors in addition to oxygen, including fumarate, nitrate, nitrite, tetrathionate and N- or S-oxides. Several novel enzymes participate in these electron transport chains, including a tungsten containing formate dehydrogenase, a Complex I that uses flavodoxin and not NADH, a periplasmic facing fumarate reductase and a cytochrome c tetrathionate reductase. This review presents an updated description of the composition and bioenergetics of these various respiratory chains as they are currently understood, including recent work that gives new insights into energy conservation during electron transport to various alternative electron acceptors. The regulation of synthesis and assembly of the electron transport chains is also discussed. A deeper appreciation of the unique features of the respiratory systems of C. jejuni may be helpful in informing strategies to control this important pathogen.

The periplasmic reduction of the electron acceptors nitrate (Em +420 mV) and trimethylamine-N-oxide (TMAO; Em +130 mV) by Nap and Tor reductases is widespread in Gram-negative bacteria and is usually considered to be driven by non-energy conserving quinol dehydrogenases. The EpsilonproteobacteriumCampylobacter jejuni can grow by nitrate and TMAO respiration and it has previously been assumed that these alternative pathways of electron transport are independent of the proton-motive menaquinol-cytochrome c reductase complex (QcrABC) that functions in oxygen-linked respiration. Here, we show that a qcrABC deletion mutant is completely deficient in oxygen-limited growth on both nitrate and TMAO and is unable to reduce these oxidants with physiological electron donors. As expected, the mutant grows normally on fumarate under oxygen-limited conditions. Thus, the periplasmic Nap and Tor reductases receive their electrons via QcrABC in C. jejuni, explaining the general absence of NapC and TorC quinol dehydrogenases in Epsilonproteobacteria. Moreover, the specific use of menaquinol (Em −75 mV) coupled with a Qcr complex to drive reduction of nitrate or TMAO against the proton-motive force allows the process to be electrogenic with a H+/2e− ratio of 2. The results have general implications for the role of Qcr complexes in bacterial oxygen-independent respiration and growth.

MicroRNAs (miRs) are small conserved RNA that regulate gene expression. Bioinformatic analysis of miRNA profiles during mouse lung development indicated a role for multiple miRNA, including miRNA-489. miR-489 increased on completion of alveolar septation [postnatal day 42 (P42)], associated with decreases in its conserved target genes insulin-like growth factor-1 ( Igf1) and tenascin C ( Tnc). We hypothesized that dysregulation of miR-489 and its target genes Igf1 and Tnc contribute to hyperoxia-induced abnormal lung development. C57BL/6 mice were exposed to normoxia (21%) or hyperoxia (85% O2) from P4 to P14, in combination with intranasal locked nucleic acid against miR-489 to inhibit miR-489, cytomegalovirus promoter (pCMV)-miR-489 to overexpress miR-489, or empty vector. Hyperoxia reduced miR-489 and increased Igf1 and Tnc. Locked nucleic acid against miR-489 improved lung development during hyperoxia and did not alter it during normoxia, whereas miR-489 overexpression inhibited lung development during normoxia. The 3′ untranslated region in vitro reporter studies confirmed Igf1 and Tnc as targets of miR-489. While miR-489 was of epithelial origin and present in exosomes, its targets Igf1 and Tnc were produced by fibroblasts. Infants with bronchopulmonary dysplasia (BPD) had reduced lung miR-489 and increased Igf1 and Tnc compared with normal preterm or term infants. These results suggest increased miR-489 is an inhibitor of alveolar septation. During hyperoxia or BPD, reduced miR-489 and increased Igf1 and Tnc may be inadequate attempts at compensation. Further inhibition of miR-489 may permit alveolar septation to proceed. The use of specific miRNA antagonists or agonists may be a therapeutic strategy for inhibited alveolarization, such as in BPD.

Contains fulltext : 168023.pdf (Publisher’s version ) (Closed access) Alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV) is a lethal lung developmental disorder caused by heterozygous point mutations or genomic deletion copy-number variants (CNVs) of FOXF1 or its upstream enhancer involving fetal lung-expressed long noncoding RNA genes LINC01081 and LINC01082. Using custom-designed array comparative genomic hybridization, Sanger sequencing, whole exome sequencing (WES), and bioinformatic analyses, we studied 22 new unrelated families (20 postnatal and two prenatal) with clinically diagnosed ACDMPV. We describe novel deletion CNVs at the FOXF1 locus in 13 unrelated ACDMPV patients. Together with the previously reported cases, all 31 genomic deletions in 16q24.1, pathogenic for ACDMPV, for which parental origin was determined, arose de novo with 30 of them occurring on the maternally inherited chromosome 16, strongly implicating genomic imprinting of the FOXF1 locus in human lungs. Surprisingly, we have also identified four ACDMPV families with the pathogenic variants in the FOXF1 locus that arose on paternal chromosome 16. Interestingly, a combination of the severe cardiac defects, including hypoplastic left heart, and single umbilical artery were observed only in children with deletion CNVs involving FOXF1 and its upstream enhancer. Our data demonstrate that genomic imprinting at 16q24.1 plays an important role in variable ACDMPV manifestation likely through long-range regulation of FOXF1 expression, and may be also responsible for key phenotypic features of maternal uniparental disomy 16. Moreover, in one family, WES revealed a de novo missense variant in ESRP1, potentially implicating FGF signaling in the etiology of ACDMPV.

Campylobacter jejuni, the leading cause of human bacterial gastroenteritis, requires low environmental oxygen and high carbon dioxide for optimum growth, but the molecular basis for the carbon dioxide requirement is unclear. One factor may be inefficient conversion of gaseous CO2 to bicarbonate, the required substrate of various carboxylases. Two putative carbonic anhydrases (CAs) are encoded in the genome of C. jejuni strain NCTC 11168 (Cj0229 and Cj0237). Here, we show that the deletion of the cj0237 (canB) gene alone prevents growth in complex media at low (1% v/v) CO2 and significantly reduces the growth rate at high (5% v/v) CO2. In minimal media incubated under high CO2, the canB mutant grew on L-aspartate but not on the key C3 compounds L-serine, pyruvate and L-lactate, showing that CanB is crucial in bicarbonate provision for pyruvate carboxylase-mediated oxaloacetate synthesis. Nevertheless, purified CanB (a dimeric, anion and acetazolamide sensitive, zinc-containing type II beta-class enzyme) hydrates CO2 actively only above pH 8 and with a high Km (∼ 34 mM). At typical cytoplasmic pH values and low CO2, these kinetic properties might limit intracellular bicarbonate availability. Taken together, our data suggest CanB is a major contributor to the capnophilic growth phenotype of C. jejuni.