bluuebunny/crossref_metadata_2025 huggingface dataset:
The Drosophila melanogaster importin α3 Locus Encodes an Essential Gene Required for the Development of Both Larval and Adult Tissues
Authors: Mason D Adam , Máthé Endre , Fleming Robert J , Goldfarb David S
Published: April 2021

The nuclear transport of classical nuclear localization signal (cNLS)-containing proteins is mediated by the cNLS receptor importin α. The conventional importin α gene family in metazoan animals is composed of three clades that are conserved between flies and mammals and are referred to here as α1, α2, and α3. In contrast, plants and fungi contain only α1 genes. In this study we report that Drosophila importin α3 is required for the development of both larval and adult tissues. Importin α3 mutant flies die around the transition from first to second instar larvae, and homozygous importin α3 mutant eyes are defective. The transition to second instar larvae was rescued with importin α1, α2, or α3 transgenes, indicating that Importin α3 is normally required at this stage for an activity shared by all three importin α's. In contrast, an α3-specific biochemical activity(s) of Importin α3 is probably required for development to adults and photoreceptor cell development, since only an importin α3 transgene rescued these processes. These results are consistent with the view that the importin α's have both overlapping and distinct functions and that their role in animal development involves the spatial and temporal control of their expression.
Wolbachia and Cytoplasmic Incompatibility in the CaliforniaCulex pipiensMosquito Species Complex: Parameter Estimates and Infection Dynamics in Natural Populations
Authors: Rasgon Jason L , Scott Thomas W
Published: April 2021

Before maternally inherited bacterial symbionts like Wolbachia, which cause cytoplasmic incompatibility (CI; reduced hatch rate) when infected males mate with uninfected females, can be used in a program to control vector-borne diseases it is essential to understand their dynamics of infection in natural arthropod vector populations. Our study had four goals: (1) quantify the number of Wolbachia strains circulating in the California Culex pipiens species complex, (2) investigate Wolbachia infection frequencies and distribution in natural California populations, (3) estimate the parameters that govern Wolbachia spread among Cx. pipiens under laboratory and field conditions, and (4) use these values to estimate equilibrium levels and compare predicted infection prevalence levels to those observed in nature. Strain-specific PCR, wsp gene sequencing, and crossing experiments indicated that a single Wolbachia strain infects Californian Cx. pipiens. Infection frequency was near or at fixation in all populations sampled for 2 years along a >1000-km north-south transect. The combined statewide infection frequency was 99.4%. Incompatible crosses were 100% sterile under laboratory and field conditions. Sterility decreased negligibly with male age in the laboratory. Infection had no significant effect on female fecundity under laboratory or field conditions. Vertical transmission was >99% in the laboratory and ∼98.6% in the field. Using field data, models predicted that Wolbachia will spread to fixation if infection exceeds an unstable equilibrium point above 1.4%. Our estimates accurately predicted infection frequencies in natural populations. If certain technical hurdles can be overcome, our data indicate that Wolbachia can invade vector populations as part of an applied transgenic strategy for vector-borne disease reduction.
Genetic Structure and Diversity Among Maize Inbred Lines as Inferred From DNA Microsatellites
Authors: Liu Kejun , Goodman Major , Muse Spencer , Smith J Stephen , Buckler Ed , Doebley John
Published: April 2021

Two hundred and sixty maize inbred lines, representative of the genetic diversity among essentially all public lines of importance to temperate breeding and many important tropical and subtropical lines, were assayed for polymorphism at 94 microsatellite loci. The 2039 alleles identified served as raw data for estimating genetic structure and diversity. A model-based clustering analysis placed the inbred lines in five clusters that correspond to major breeding groups plus a set of lines showing evidence of mixed origins. A “phylogenetic” tree was constructed to further assess the genetic structure of maize inbreds, showing good agreement with the pedigree information and the cluster analysis. Tropical and subtropical inbreds possess a greater number of alleles and greater gene diversity than their temperate counterparts. The temperate Stiff Stalk lines are on average the most divergent from all other inbred groups. Comparison of diversity in equivalent samples of inbreds and open-pollinated landraces revealed that maize inbreds capture <80% of the alleles in the landraces, suggesting that landraces can provide additional genetic diversity for maize breeding. The contributions of four different segments of the landrace gene pool to each inbred group's gene pool were estimated using a novel likelihood-based model. The estimates are largely consistent with known histories of the inbreds and indicate that tropical highland germplasm is poorly represented in maize inbreds. Core sets of inbreds that capture maximal allelic richness were defined. These or similar core sets can be used for a variety of genetic applications in maize.
Recombination Can Evolve in Large Finite Populations Given Selection on Sufficient Loci
Authors: Iles Mark M , Walters Kevin , Cannings Chris
Published: April 2021

It is well known that an allele causing increased recombination is expected to proliferate as a result of genetic drift in a finite population undergoing selection, without requiring other mechanisms. This is supported by recent simulations apparently demonstrating that, in small populations, drift is more important than epistasis in increasing recombination, with this effect disappearing in larger finite populations. However, recent experimental evidence finds a greater advantage for recombination in larger populations. These results are reconciled by demonstrating through simulation without epistasis that for m loci recombination has an appreciable selective advantage over a range of population sizes (am, bm). bm increases steadily with m while am remains fairly static. Thus, however large the finite population, if selection acts on sufficiently many loci, an allele that increases recombination is selected for. We show that as selection acts on our finite population, recombination increases the variance in expected log fitness, causing indirect selection on a recombination-modifying locus. This effect is enhanced in those populations with more loci because the variance in phenotypic fitnesses in relation to the possible range will be smaller. Thus fixation of a particular haplotype is less likely to occur, increasing the advantage of recombination.
Distinct Signatures for Mutator Sensitivity of lacZ Reversions and for the Spectrum of lacI/lacO Forward Mutations on the Chromosome of Nondividing Escherichia coli
Authors: Bharatan Shanti M , Reddy Manjula , Gowrishankar J
Published: April 2021

A conditional lethal galE(Ts)-based strategy was employed in Escherichia coli, first to eliminate all growth-associated chromosomal reversions in lacZ or forward mutations in lacI/lacO by incubation at the restrictive temperature and subsequently to recover (as papillae) spontaneous mutations that had arisen in the population of nondividing cells after shift to the permissive temperature. Data from lacZ reversion studies in mutator strains indicated that the products of all genes for mismatch repair (mutHLS, dam, uvrD), of some for oxidative damage repair (mutMT), and of that for polymerase proofreading (dnaQ) are required in dividing cells; some others for oxidative damage repair (mutY, nth nei) are required in both dividing and nondividing cells; and those for alkylation damage repair (ada ogt) are required in nondividing cells. The spectrum of lacI/lacO mutations in nondividing cells was distinguished both by lower frequencies of deletions and IS1 insertions and by the unique occurrence of GC-to-AT transitions at lacO +5. In the second approach to study mutations that had occurred in nondividing cells, lacI/lacO mutants were selected as late-arising papillae from the lawn of a galE+ strain; once again, transitions at lacO +5 were detected among the mutants that had been obtained from populations initially grown on poor carbon sources such as acetate, palmitate, or succinate. Our results indicate that the lacO +5 site is mutable only in nondividing cells, one possible mechanism for which might be that random endogenous alkylation (or oxidative) damage to DNA in these cells is efficiently corrected by the Ada Ogt (or Nth Nei) repair enzymes at most sites but not at lacO +5. Furthermore, the late-arising papillae from the second approach were composed almost exclusively of dominant lacI/lacO mutants. This finding lends support to “instantaneous gratification” models in which a spontaneous lesion, occurring at a random site in DNA of a nondividing cell, is most likely to be fixed as a mutation if it allows the cell to immediately exit the nondividing state.
Reciprocal Translocations in Saccharomyces cerevisiae Formed by Nonhomologous End Joining
Authors: Yu Xin , Gabriel Abram
Published: April 2021

Reciprocal translocations are common in cancer cells, but their creation is poorly understood. We have developed an assay system in Saccharomyces cerevisiae to study reciprocal translocation formation in the absence of homology. We induce two specific double-strand breaks (DSBs) simultaneously on separate chromosomes with HO endonuclease and analyze the subsequent chromosomal rearrangements among surviving cells. Under these conditions, reciprocal translocations via nonhomologous end joining (NHEJ) occur at frequencies of ∼2-7 × 10-5/cell exposed to the DSBs. Yku80p is a component of the cell’s NHEJ machinery. In its absence, reciprocal translocations still occur, but the junctions are associated with deletions and extended overlapping sequences. After induction of a single DSB, translocations and inversions are recovered in wild-type and rad52 strains. In these rearrangements, a nonrandom assortment of sites have fused to the DSB, and their junctions show typical signs of NHEJ. The sites tend to be between open reading frames or within Ty1 LTRs. In some cases the translocation partner is formed by a break at a cryptic HO recognition site. Our results demonstrate that NHEJ-mediated reciprocal translocations can form in S. cerevisiae as a consequence of DSB repair.
Genetic Complexity Underlying Hybrid Male Sterility in Drosophila
Authors: Sawamura Kyoichi , Roote John , Wu Chung-I , Yamamoto Masa-Toshi
Published: April 2021

Recent genetic analyses of closely related species of Drosophila have indicated that hybrid male sterility is the consequence of highly complex synergistic effects among multiple genes, both conspecific and heterospecific. On the contrary, much evidence suggests the presence of major genes causing hybrid female sterility and inviability in the less-related species, D. melanogaster and D. simulans. Does this contrast reflect the genetic distance between species? Or, generally, is the genetic basis of hybrid male sterility more complex than that of hybrid female sterility and inviability? To clarify this point, the D. simulans introgression of the cytological region 34D-36A to the D. melanogaster genome, which causes recessive male sterility, was dissected by recombination, deficiency, and complementation mapping. The 450-kb region between two genes, Suppressor of Hairless and snail, exhibited a strong effect on the sterility. Males are (semi-)sterile if this region of the introgression is made homozygous or hemizygous. But no genes in the region singly cause the sterility; this region has at least two genes, which in combination result in male sterility. Further, the males are less fertile when heterozygous with a larger introgression, which suggests that dominant modifiers enhance the effects of recessive genes of male sterility. Such an epistatic view, even in the less-related species, suggests that the genetic complexity is special to hybrid male sterility.
On the Rate and Linearity of Viability Declines in Drosophila Mutation-Accumulation Experiments: Genomic Mutation Rates and Synergistic Epistasis Revisited
Authors: Fry James D
Published: April 2021

High rates of deleterious mutations could severely reduce the fitness of populations, even endangering their persistence; these effects would be mitigated if mutations synergize each others’ effects. An experiment by Mukai in the 1960s gave evidence that in Drosophila melanogaster, viability-depressing mutations occur at the surprisingly high rate of around one per zygote and that the mutations interact synergistically. A later experiment by Ohnishi seemed to support the high mutation rate, but gave no evidence for synergistic epistasis. Both of these studies, however, were flawed by the lack of suitable controls for assessing viability declines of the mutation-accumulation (MA) lines. By comparing homozygous viability of the MA lines to simultaneously estimated heterozygous viability and using estimates of the dominance of mutations in the experiments, I estimate the viability declines relative to an appropriate control. This approach yields two unexpected conclusions. First, in Ohnishi’s experiment as well as in Mukai’s, MA lines showed faster-than-linear declines in viability, indicative of synergistic epistasis. Second, while Mukai’s estimate of the genomic mutation rate is supported, that from Ohnishi’s experiment is an order of magnitude lower. The different results of the experiments most likely resulted from differences in the starting genotypes; even within Mukai’s experiment, a subset of MA lines, which I argue probably resulted from a contamination event, showed much slower viability declines than did the majority of lines. Because different genotypes may show very different mutational behavior, only studies using many founding genotypes can determine the average rate and distribution of effects of mutations relevant to natural populations.
EXO1 Plays a Role in Generating Type I and Type II Survivors in Budding Yeast
Authors: Maringele Laura , Lydall David
Published: April 2021

Telomerase-defective budding yeast cells escape senescence by using homologous recombination to amplify telomeric or subtelomeric structures. Similarly, human cells that enter senescence can use homologous recombination for telomere maintenance, when telomerase cannot be activated. Although recombination proteins required to generate telomerase-independent survivors have been intensively studied, little is known about the nucleases that generate the substrates for recombination. Here we demonstrate that the Exo1 exonuclease is an initiator of the recombination process that allows cells to escape senescence and become immortal in the absence of telomerase. We show that EXO1 is important for generating type I survivors in yku70Δ mre11Δ cells and type II survivors in tlc1Δ cells. Moreover, in tlc1Δ cells, EXO1 seems to contribute to the senescence process itself.
Role of the Nuclease Activity of Saccharomyces cerevisiae Mre11 in Repair of DNA Double-Strand Breaks in Mitotic Cells
Authors: Lewis L Kevin , Storici Francesca , Van Komen Stephen , Calero Shanna , Sung Patrick , Resnick Michael A
Published: April 2021

The Rad50:Mre11:Xrs2 (RMX) complex functions in repair of DNA double-strand breaks (DSBs) by recombination and nonhomologous end-joining (NHEJ) and is also required for telomere stability. The Mre11 subunit exhibits nuclease activities in vitro, but the role of these activities in repair in mitotic cells has not been established. In this study we have performed a comparative study of three mutants (mre11-D16A, -D56N, and -H125N) previously shown to have reduced nuclease activities in vitro. In ends-in and ends-out chromosome recombination assays using defined plasmid and oligonucleotide DNA substrates, mre11-D16A cells were as deficient as mre11 null strains, but defects were small in mre11-D56N and -H125N mutants. mre11-D16A cells, but not the other mutants, also displayed strong sensitivity to ionizing radiation, with residual resistance largely dependent on the presence of the partially redundant nuclease Exo1. mre11-D16A mutants were also most sensitive to the S-phase-dependent clastogens hydroxyurea and methyl methanesulfonate but, as previously observed for D56N and H125N mutants, were not defective in NHEJ. Importantly, the affinity of purified Mre11-D16A protein for Rad50 and Xrs2 was indistinguishable from wild type and the mutant protein formed complexes with equivalent stoichiometry. Although the role of the nuclease activity has been questioned in previous studies, the comparative data presented here suggest that the nuclease function of Mre11 is required for RMX-mediated recombinational repair and telomere stabilization in mitotic cells.
Quantitative Trait Loci Affecting Starvation Resistance in Drosophila melanogaster
Authors: Harbison Susan T , Yamamoto Akihiko H , Fanara Juan J , Norga Koenraad K , Mackay Trudy F C
Published: April 2021

The ability to withstand periods of scarce food resources is an important fitness trait. Starvation resistance is a quantitative trait controlled by multiple interacting genes and exhibits considerable genetic variation in natural populations. This genetic variation could be maintained in the face of strong selection due to a trade-off in resource allocation between reproductive activity and individual survival. Knowledge of the genes affecting starvation tolerance and the subset of genes that affect variation in starvation resistance in natural populations would enable us to evaluate this hypothesis from a quantitative genetic perspective. We screened 933 co-isogenic P-element insertion lines to identify candidate genes affecting starvation tolerance. A total of 383 P-element insertions induced highly significant and often sex-specific mutational variance in starvation resistance. We also used deficiency complementation mapping followed by complementation to mutations to identify 12 genes contributing to variation in starvation resistance between two wild-type strains. The genes we identified are involved in oogenesis, metabolism, and feeding behaviors, indicating a possible link to reproduction and survival. However, we also found genes with cell fate specification and cell proliferation phenotypes, which implies that resource allocation during development and at the cellular level may also influence the phenotypic response to starvation.
Nucleotide Variation in thetinmanandbagpipeHomeobox Genes ofDrosophila melanogaster
Authors: Balakirev Evgeniy S , Ayala Francisco J
Published: April 2021

The tinman (tin) and bagpipe (bap) genes are members of the NK homeobox gene family of Drosophila, so that tin occupies a higher position than bap in the regulatory hierarchy. Little is known about the level and pattern of genetic polymorphism in homeobox genes. We have analyzed nucleotide polymorphism in 27 strains of Drosophila melanogaster and one each of D. simulans and D. sechellia, within two closely linked regions encompassing a partial sequence of tin and the complete sequence of bap. The two genes exhibit different levels and patterns of nucleotide diversity. Two sets of sharply divergent sequence types are detected for tin. The haplotype structure of bap is more complex: about half of the sequences are identical (or virtually so), while the rest are fairly heterogeneous. The level of silent nucleotide variability is 0.0063 for tin but significantly higher, 0.0141, for bap, a level of polymorphism comparable to the most polymorphic structural genes of D. melanogaster. Recombination rate and gene conversion are also higher for bap than for tin. There is strong linkage disequilibrium, with the highest values in the introns of both genes and exon II of bap. The patterns of polymorphism in tin and bap are not compatible with an equilibrium model of selective neutrality. We suggest that negative selection and demographic history are the major factors shaping the pattern of nucleotide polymorphism in the tin and bap genes; moreover, there are clear indications of positive selection in the bap gene.
Evolution ofDMY, a Newly Emergent Male Sex-Determination Gene of Medaka Fish
Authors: Zhang Jianzhi
Published: April 2021

The Japanese medaka fish Oryzias latipes has an XX/XY sex-determination system. The Y-linked sex-determination gene DMY is a duplicate of the autosomal gene DMRT1, which encodes a DM-domain-containing transcriptional factor. DMY appears to have originated recently within Oryzias, allowing a detailed evolutionary study of the initial steps that led to the new gene and new sex-determination system. Here I analyze the publicly available DMRT1 and DMY gene sequences of Oryzias species and report the following findings. First, the synonymous substitution rate in DMY is 1.73 times that in DMRT1, consistent with the male-driven evolution hypothesis. Second, the ratio of the rate of nonsynonymous nucleotide substitution (dN) to that of synonymous substitution (dS) is significantly higher in DMY than in DMRT1. Third, in DMRT1, the dN/dS ratio for the DM domain is lower than that for non-DM regions, as expected from the functional importance of the DM domain. But in DMY, the opposite is observed and the DM domain is likely under positive Darwinian selection. Fourth, only one characteristic amino acid distinguishes all DMY sequences from all DMRT1 sequences, suggesting that a single amino acid change may be largely responsible for the establishment of DMY as the male sex-determination gene in medaka fish.
Spatial and temporal variability in somatic growth in fisheries stock assessment models: evaluating the consequences of misspecification
Authors: Correa Giancarlo M , McGilliard Carey R , Ciannelli Lorenzo , Fuentes Claudio
Published: April 2021

Distinct types of fish species experience variation in somatic growth rates over their life span; however, growth has historically been assumed to be invariant across time and space in integrated analysis-based stock assessment. A few previous studies have reported biased and imprecise assessment model outcomes when variability in somatic growth was ignored. In this study, we used a simulation-estimation framework to expand previous analyses and to examine the consequences of ignoring or incorporating spatial and temporal (year- and cohort-specific) variability in somatic growth in stock assessment models. The study included three life history types: small pelagic (e.g. sardine), gadids (e.g. cod), and long-lived (e.g. rockfish). In general, ignoring any type of variability in somatic growth led to biased and imprecise estimates of stock spawning biomass and management quantities. Unequal distribution of fishing mortality across space had large impacts on the performance of estimation models as well. Conversely, accounting for somatic growth variability, either by including an environmental index, estimating annual deviates, or implementing a spatially explicit model, produced unbiased and precise results. This study shows that somatic growth variability might produce large effects in stock assessments when ignored and provides pertinent information for stock assessment best practice guidelines.
An unfitted Eulerian finite element method for the time-dependent Stokes problem on moving domains
Authors: von Wahl Henry , Richter Thomas , Lehrenfeld Christoph
Published: April 2021

We analyse a Eulerian finite element method, combining a Eulerian time-stepping scheme applied to the time-dependent Stokes equations with the CutFEM approach using inf-sup stable Taylor–Hood elements for the spatial discretization. This is based on the method introduced by Lehrenfeld & Olshanskii (2019, A Eulerian finite element method for PDEs in time-dependent domains. ESAIM: M2AN, 53, 585–614) in the context of a scalar convection–diffusion problems on moving domains, and extended to the nonstationary Stokes problem on moving domains by Burman et al. (2019, arXiv:1910.03054 [math.NA]) using stabilized equal-order elements. The analysis includes the geometrical error made by integrating over approximated level set domains in the discrete CutFEM setting. The method is implemented and the theoretical results are illustrated using numerical examples.
Is the German Nuclear strategy lawful under EU law? Article 194(2) TFEU and its limitations
Authors: Rochette Gustavo , Vela Elena
Published: April 2021

Germany streamlined its nuclear phase-out policy following the Fukushima Daishi nuclear accident, planning to decommission all of its nuclear capacity by 2022. As a member of the EU, although Germany is entitled to determine its own energy mix pursuant to Article 194(2) of the TFEU, this policy may conflict with other EU legislation and jurisprudence relevant to the EU energy sector. In particular, in this paper, we argue that the German nuclear phase-out may be contrary to the European Atomic Energy Community Treaty and the primary and secondary EU legislation regarding security of energy supply. We explore how Germany’s commitments under this legislation act as a limitation to its national energy policy autonomy and might entail the unlawfulness of the German nuclear phase-out strategy.
Non-glandular trichomes of sunflower are important in the absorption and translocation of foliar-applied Zn
Authors: Li Cui , Wu Jingtao , Blamey F Pax C , Wang Linlin , Zhou Lina , Paterson David J , van der Ent Antony , Fernández Victoria , Lombi Enzo , Wang Yuheng , Kopittke Peter M
Published: April 2021

Trichomes are potentially important for absorption of foliar fertilizers. A study has shown that the non-glandular trichromes (NGTs) of sunflower (Helianthus annuus) accumulated high concentrations of foliar-applied zinc (Zn); however, the mechanisms of Zn accumulation in the NGTs and the fate of this Zn are unclear. Here we investigated how foliar-applied Zn accumulates in the NGTs and the subsequent translocation of this Zn. Time-resolved synchrotron-based X-ray fluorescence microscopy and transcriptional analyses were used to probe the movement of Zn in the NGTs, with the cuticle composition of the NGTs examined using confocal Raman microscopy. The accumulation of Zn in the NGTs is both an initial preferential absorption process and a subsequent translocation process. This preferred absorption is likely because the NGT base has a higher hydrophilicity, whilst the subsequent translocation is due to the presence of plasmodesmata, Zn-chelating ligands, and Zn transporters in the NGTs. Furthermore, the Zn sequestered in the NGTs was eventually translocated out of the trichome once the leaf Zn concentration had decreased, suggesting that the NGTs are also important in maintaining leaf Zn homeostasis. This study demonstrates for the first time that trichomes have a key structural and functional role in the absorption and translocation of foliar-applied Zn.
Overexpression of thioredoxin m in chloroplasts alters carbon and nitrogen partitioning in tobacco
Authors: Ancín María , Larraya Luis , Florez-Sarasa Igor , Bénard Camille , Fernández-San Millán Alicia , Veramendi Jon , Gibon Yves , Fernie Alisdair R , Aranjuelo Iker , Farran Inmaculada
Published: April 2021

In plants, there is a complex interaction between carbon (C) and nitrogen (N) metabolism, and its coordination is fundamental for plant growth and development. Here, we studied the influence of thioredoxin (Trx) m on C and N partitioning using tobacco plants overexpressing Trx m from the chloroplast genome. The transgenic plants showed altered metabolism of C (lower leaf starch and soluble sugar accumulation) and N (with higher amounts of amino acids and soluble protein), which pointed to an activation of N metabolism at the expense of carbohydrates. To further delineate the effect of Trx m overexpression, metabolomic and enzymatic analyses were performed on these plants. These results showed an up-regulation of the glutamine synthetase–glutamate synthase pathway; specifically tobacco plants overexpressing Trx m displayed increased activity and stability of glutamine synthetase. Moreover, higher photorespiration and nitrate accumulation were observed in these plants relative to untransformed control plants, indicating that overexpression of Trx m favors the photorespiratory N cycle rather than primary nitrate assimilation. Taken together, our results reveal the importance of Trx m as a molecular mediator of N metabolism in plant chloroplasts.
Global Patterns and Prevalence of Dual and Poly-Tobacco Use: A Systematic Review
Authors: Chen Daniel Tzu-Hsuan , Girvalaki Charis , Mechili Enkeleint A , Millett Christopher , Filippidis Filippos T
Published: April 2021

Introduction

Improving understanding of the epidemiology of dual and poly-tobacco product use is essential for tobacco control policy and practice. The present study aimed to systematically review existing epidemiologic evidence on current dual and poly-tobacco use among adults globally.

Methods

We systematically searched online databases for studies published up to June 30, 2020. We included quantitative studies with measures of nationally representative prevalence of current dual or poly-tobacco use among adults. Prevalence estimates for each country were extracted manually and stratified by WHO regions and World Bank income classifications.

Results

Twenty studies with nationally representative prevalence data on current dual or poly-tobacco use in the adult population across 48 countries were included. Definitions of dual and poly-tobacco use varied widely. Prevalence of dual and poly-tobacco use was higher in low- and lower-middle-income countries compared to other higher-income countries. Current dual use of smoked and smokeless tobacco products among males ranged from 0.2% in Ukraine (2010) and Mexico (2009) to 17.9% in Nepal (2011). Poly-tobacco use among males ranged from 0.8% in Mexico (2009) and 0.9% in Argentina (2010) to 11.4% in the United Kingdom and 11.9% in Denmark in 2012. Dual tobacco use was generally higher in South-East Asia; poly-tobacco use was prevalent in Europe as well as in South-East Asia.

Conclusions

This is the first systematic review of the prevalence estimates of dual and poly-tobacco use among adults globally. The results of the current study could significantly help health policy makers to implement effective tobacco control policies.

Implications

This study demonstrates that dual/poly-tobacco use is common in many countries of the world, and highlights the need for in-depth exploration of this field in future studies, especially in high prevalence regions such as South-East Asian and European countries. In light of this, the global tobacco control community and health authorities should also agree upon a consistent operational definition of dual and poly-tobacco use to propel research and improve surveillance of dual/poly-use in health surveys for better communication and understanding of these phenomena.
Bm-miR172c-5p Regulates Lignin Biosynthesis and Secondary Xylem Thickness by Altering the Ferulate 5 Hydroxylase Gene in Bacopa monnieri
Authors: Jeena Gajendra Singh , Joshi Ashutosh , Shukla Rakesh Kumar
Published: April 2021

MicroRNAs (miRNAs) are small non-coding, endogenous RNAs containing 20–24 nucleotides that regulate the expression of target genes involved in various plant processes. A total of 1,429 conserved miRNAs belonging to 95 conserved miRNA families and 12 novel miRNAs were identified from Bacopa monnieri using small RNA sequencing. The Bm-miRNA target transcripts related to the secondary metabolism were further selected for validation. The Bm-miRNA expression in shoot and root tissues was negatively correlated with their target transcripts. The Bm-miRNA cleavage sites were mapped within the coding or untranslated region as depicted by the modified RLM-RACE. In the present study, we validate three miRNA targets, including asparagine synthetase, cycloartenol synthase and ferulate 5 hydroxylase (F5H) and elucidate the regulatory role of Bm-miR172c-5p, which cleaves the F5H gene involved in the lignin biosynthesis. Overexpression (OE) of Bm-miR172c-5p precursor in B. monnieri suppresses F5H gene, leading to reduced lignification and secondary xylem thickness under control and drought stress. By contrast, OE of endogenous target mimics (eTMs) showed enhanced lignification and secondary xylem thickness leading to better physiological response under drought stress. Taken together, we suggest that Bm-miRNA172c-5p might be a key player in maintaining the native phenotype of B. monnieri under control and different environmental conditions.
Patient-Reported Outcome–Based Quality Indicators in Dutch Primary Care Physical Therapy for Patients With Nonspecific Low Back Pain: A Cohort Study
Authors: Verburg Arie C , van Dulmen Simone A , Kiers Henri , Nijhuis-van der Sanden Maria W G , van der Wees Philip J
Published: April 2021

Objective

The purpose of this study was to define and select a core set of outcome-based quality indicators, accepted by stakeholders on usability and perceived added value as a quality improvement tool, and to formulate recommendations for the next implementation step.

Methods

In phase 1, 15 potential quality indicators were defined for patient-reported outcome measures and associated domains, namely the Numeric Pain Rating Scale (NPRS) for pain intensity, the Patient Specific Functioning Scale (PSFS) for physical activity, the Quebec Back Pain Disability Scale for physical functioning, and the Global Perceived Effect—Dutch Version for perceived effect. Their comparability and discriminatory characteristics were described using cohort data. In phase 2, a core set of quality indicators was selected based on consensus among stakeholders in focus group meetings.

Results

In total, 65,815 completed treatment episodes for patients with nonspecific low back pain were provided by 1009 physical therapists from 219 physical therapist practices. The discriminability between physical therapists of all potential 15 quality indicators was adequate, with intraclass correlation coefficients between 0.08 and 0.30. Stakeholders selected a final core set of 6 quality indicators: 2 process indicators (the routine measurement of NPRS and the PSFS) and 4 outcome indicators (pretreatment and posttreatment change scores for the NPRS, PSFS, Quebec Back Pain Disability Scale, and the minimal clinically important difference of the Global Perceived Effect—Dutch Version).

Conclusion

This study described and selected a core set of outcome-based quality indicators for physical therapy in patients with nonspecific low back pain. The set was accepted by stakeholders for having added value for daily practice in physical therapy primary care and was found useful for quality improvement initiatives. Further studies need to focus on improvement of using the core set of outcome-based quality indicators as a quality monitoring and evaluation instrument.

Impact

Patient-reported outcome–based quality indicators developed from routinely collected clinical data are promising for use in quality improvement in daily practice.
COVID-19 and Territorial Policy Dynamics in Western Europe: Comparing France, Spain, Italy, Germany, and the United Kingdom
Authors: Vampa Davide
Published: April 2021

This article seeks to assess and explain territorial policy dynamics in five European countries—Italy, Spain, Germany, France and the United Kingdom—from the start of the COVID-19 pandemic up to early 2021. The crisis has clearly highlighted well-known differences between centralized and decentralized systems. Yet focusing on this dichotomy is not sufficient. It is suggested that, while the distribution of authority between central and regional governments matters, policy dynamics—that is, how different territorial levels interact in policy-making processes—are even more important in driving multi-level responses to the emergency. Whether these dynamics are hierarchical (France), competitive (Italy and Spain), cooperative (Germany) or mixed (the United Kingdom) depends on how pre-crisis institutional, sectoral and political “causal forces” moderate the impact of an exogenous shock.
Toxicokinetic and Genotoxicity Study of NNK in Male Sprague Dawley Rats Following Nose-Only Inhalation Exposure, Intraperitoneal Injection, and Oral Gavage
Authors: Hu Shu-Chieh , Bryant Matthew S , Sepehr Estatira , Kang Hyun-Ki , Trbojevich Raul , Lagaud Guy , Mehta Darshan , Ding Wei , Mittelstaedt Roberta A , Pearce Mason G , Bishop Michelle E , Davis Kelly J , Lewis Sherry M , Chemerynski Susan , Yee Steven B , Coraggio Melis , Rosenfeldt Hans , Yeager R Philip , Howard Paul C , Tang Yunan
Published: April 2021

The tobacco-specific nitrosamine NNK [4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone] is found in tobacco products and tobacco smoke. NNK is a potent genotoxin and human lung carcinogen; however, there are limited inhalation data for the toxicokinetics (TK) and genotoxicity of NNK in vivo. In the present study, a single dose of 5 × 10−5, 5 × 10−3, 0.1, or 50 mg/kg body weight (BW) of NNK, 75% propylene glycol (vehicle control), or air (sham control) was administered to male Sprague-Dawley (SD) rats (9–10 weeks age) via nose-only inhalation (INH) exposure for 1 h. For comparison, the same doses of NNK were administered to male SD rats via intraperitoneal injection (IP) and oral gavage (PO). Plasma, urine, and tissue specimens were collected at designated time points and analyzed for levels of NNK and its major metabolite 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and tissue levels of DNA adduct O6-methylguanine by LC/MS/MS. TK data analysis was performed using a non-linear regression program. For the genotoxicity subgroup, tissues were collected at 3 h post-dosing for comet assay analysis. Overall, the TK data indicated that NNK was rapidly absorbed and metabolized extensively to NNAL after NNK administration via the three routes. The IP route had the greatest systemic exposure to NNK. NNK metabolism to NNAL appeared to be more efficient via INH than IP or PO. NNK induced significant increases in DNA damage in multiple tissues via the three routes. The results of this study provide new information and understanding of the TK and genotoxicity of NNK.
Different Polyphenolic Parenchyma Cell and Phloem Axial Resin Duct-Like Structure Formation Rates in Cupressus sempervirens Clones Infected with Seiridium cardinale
Authors: Della Rocca G. , Posarelli I. , Morandi F. , Tani C. , Barberini S. , Danti R. , Moricca S. , Papini A.
Published: April 2021

The aim of this study was the characterization of constitutive and induced defense mechanisms in the bark tissues of Cupressus sempervirens before and after infection with the bark fungus Seiridium cardinale, which is responsible for cypress canker disease. The time-course development of polyphenolic parenchyma (PP) cells and phloem axial resin duct (PARD)-like structures in the phloem was investigated in two C. sempervirens clones, one resistant and one susceptible to the disease, through anatomical and histological observations carried out by light microscope during a 19-day trial. PP cells were constitutively more abundant in the canker-resistant clone (R clone) compared with the susceptible clone (S clone), whereas PARD-like structures were not present in the bark of untreated plants of both clones. PP cells increased in both clones as a response to infection, but in the R clone, they were more abundant 5 and 12 days after inoculation. After inoculation, PARD-like structures appeared in the phloem after 5 days in the R clone and only after 12 days in the S clone. Even the number of cells surrounding the PARD-like structures was higher in the R clone 5 and 12 days after inoculation compared with the S clone. These observations demonstrate a faster phloem response of the R clone in the early phase of the infection. This may slow down initial growth of the fungus, contributing to the resistance mechanism.
First Expansion of the Public Tomato Brown Rugose Fruit Virus (ToBRFV) Nextstrain Build; Inclusion of New Genomic and Epidemiological Data
Authors: van de Vossenberg Bart T. L. H. , Dawood Thikra , Woźny Marek , Botermans Marleen
Published: April 2021

Tomato brown rugose fruit virus (ToBRFV) is a tobamovirus that was first detected in Israel and Jordan following an outbreak of a new disease infecting tomato in 2014. Since then, the virus has been reported from all continents except Oceania and Antarctica. In response to the first finding of the virus in The Netherlands, the Dutch National Plant Protection Organization created a ToBRFV Nextstrain build (v1). In this report, we announce 47 new (near) complete ToBRFV genomes and the generation of the new ToBRFV Nextstrain (v2) build containing 118 ToBRFV genomes with associated geographic and epidemiological data. Examples of utilization of the genomic sequences are presented, and we report the first sequence from South America and present a novel hypothesis on the possible ToBRFV center of origin.

[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .
Postoperative Pain Mediates the Association Between Peripheral Nerve Blocks and Postoperative Sleep Following Lower Extremity Arthroplasty
Authors: Giordano Nicholas A. , Kent Michael , Andersen Stephanie G. , Scott-Richardson Maya , Highland Krista B.
Published: April 2021

Objectives:

Despite substantial research showing the bidrectional relationship between sleep and pain, there remains a dearth of research examining the role of perioperative pain management interventions in mitigating postoperative sleep disturbances. This secondary analysis of a prospective observational multisite study examined the association between peripheral nerve block (PNB) use during total knee or total hip arthroplasty (TKA/THA) procedures and postoperative pain and sleep outcomes.

Materials and Methods:

Adult patients undergoing TKA or THA procedures were recruited from 2 tertiary care facilities. Average pain and sleep disturbance scores were collected preoperatively and at 1- and 2-week postoperatively. Participants were not randomized to receive PNB. Postoperative outcomes were compared based on receipt of PNB during surgery. Structural equation modeling path analysis was utilized to model multiple co-occurring relationships, including mediation pathways between perioperative pain management approaches, pain, and postoperative sleep outcomes.

Results:

Of the 197 participants, 53% received PNB. Mediation analyses indicated that PNB was indirectly associated with 1-week sleep disturbance via its effects on 1-week pain intensity (β=−0.02, 95% confidence interval [CI] −0.04, −0.001, P=0.04). In addition, PNB was indirectly associated with 2-week sleep disturbance, via its effects on 1-week pain intensity and 1-week sleep disturbance (β=−0.04, 95% CI −0.07, −0.02, P=0.04). Lastly, PNB was indirectly associated with 2-week pain intensity via its effects on 1-week pain intensity (β=−0.10, 95% CI −0.19, −0.02, P=0.02).

Conclusions:

Receipt of PNB during TKA or THA was found to be associated with improved 1-week postoperative pain intensity, which in turn was found to be associated with lower sleep disturbances at both 1- and 2-week postoperative time points. Multimodal opioid sparing pain management interventions, capable of improving postoperative sleep, are vital to improving recovery and rehabilitation following arthroplasty.
Diagnostic Potential of Coagulation-Related Biomarkers for Sepsis in the Emergency Department: Protocol for a Pilot Observational Cohort Study
Authors: Arora Jaskirat , Klowak Jennifer A. , Parpia Sameer , Zapata-Canivilo Marcelo , Faidi Walaa , Skappak Christopher , Gregoris Rachael , Kretz Colin A. , Dwivedi Dhruva J. , de Wit Kerstin , Welsford Michelle , Fox-Robichaud Alison
Published: April 2021

Background:

Between 75% and 80% of patients with sepsis arrive in the hospital through the emergency department. Early diagnosis is important to alter patient prognosis, but currently, there is no reliable biomarker. The innate immune response links inflammation and coagulation. Several coagulation -related biomarkers are associated with poor prognosis in the ICU. The role of coagulation biomarkers to aid in early sepsis diagnosis has not previously been investigated. The objective of our study is to determine the individual or combined accuracy of coagulation and inflammation biomarkers with standard biochemical tests to diagnose adult septic patients presenting to the emergency department.

Methods:

in the Emergency Department is a prospective, observational cohort study with a target enrolment of 250 suspected septic patients from two Canadian emergency departments. The emergency physicians will enroll patients with suspected sepsis. Blood samples will be collected at two time points (initial presentation and 4 hr following). Patients will be adjudicated into septic, infected, or not infected status in accordance with the Sepsis-3 definitions. Patient demographics, cultures, diagnosis, and biomarkers will be reported using descriptive statistics. Optimal cut off values with sensitivity and specificity for each biomarker will be determined using C-statistics to distinguish between septic and nonseptic patients. Stepwise multiple logistic regression analysis with exclusion of nonsignificant covariates from the final model will be used to establish a panel of biomarkers.

Conclusions:

Our protocol describes the processes and methods for a pragmatic observational biomarker study in the emergency department. This study will seek to determine the potential diagnostic importance of early coagulation abnormalities to identify additional tools for sepsis diagnosis.
Using Eye Tracking for Measuring Cognitive Workload During Clinical Simulations
Authors: Wilbanks Bryan A. , Aroke Edwin , Dudding Katherine M.
Published: April 2021

High-fidelity clinical simulations can be used by clinicians to acquire technical (physical ability and knowledge) and non-technical (cognitive and social processes) skills. Excessive cognitive workload contributes to medical errors because of the impact on both technical and non-technical skills. Many studies measure cognitive workload with psychometric instruments that limit the assessment of cognitive workload to a single time period and may involve response bias. Using eye tracking to measure task-evoked pupillary responses allows the measurement of changes in pupil diameter related to the cognitive workload associated with a specific activity. Incorporating eye tracking with high-fidelity clinical simulations provides a reliable and continuous assessment of cognitive workload. The purpose of this literature review is to summarize the use of eye-tracking technology to measure cognitive workload of healthcare providers to generate evidence-based guidelines for measuring cognitive workload during high-fidelity clinical simulations. What this manuscript adds to the body of literature is a summary of best practices related to the different methods of measuring cognitive workload, benefits and limitations of using eye tracking, and high-fidelity clinical simulation design considerations for successful integration of eye tracking.