General Relativistic effects in preheating

General relativistic effects in the form of metric perturbations are usually neglected in the preheating era that follows inflation. We argue that in realistic multi-field models these effects are in fact crucial, and the fully coupled system of metric and quantum field fluctuations needs to be considered. Metric perturbations are resonantly amplified, breaking the scale-invariance of the primordial spectrum, and in turn stimulate scalar field resonances via gravitational rescattering. This non-gravitationally dominated nonlinear growth of gravitational fluctuations may have significant effects on the Doppler peaks in the cosmic background radiation, primordial black hole formation, gravitational waves and nonthermal symmetry restoration.Comment: 10 pages. 1 .ps fig. Matches version to appear in Phys Lett

Restoring the sting to metric preheating

The relative growth of field and metric perturbations during preheating is sensitive to initial conditions set in the preceding inflationary phase. Recent work suggests this may protect super-Hubble metric perturbations from resonant amplification during preheating. We show that this possibility is fragile and sensitive to the specific form of the interactions between the inflaton and other fields. The suppression is naturally absent in two classes of preheating in which either (1) the vacua of the non-inflaton fields during inflation are deformed away from the origin, or (2) the effective masses of non-inflaton fields during inflation are small but during preheating are large. Unlike the simple toy model of a $g^2 \phi^2 \chi^2$ coupling, most realistic particle physics models contain these other features. Moreover, they generically lead to both adiabatic and isocurvature modes and non-Gaussian scars on super-Hubble scales. Large-scale coherent magnetic fields may also appear naturally.Comment: 6 pages, 3 ps figures, RevTex, revised discussion of backreaction and new figure. To appear Phys. Rev. D (Rapid Communication

Dystrophin is required for the formation of stable muscle attachments in the zebrafish embryo

A class of recessive lethal zebrafish mutations has been identified in which normal skeletal muscle differentiation is followed by a tissue-specific degeneration that is reminiscent of the human muscular dystrophies. Here, we show that one of these mutations, sapje, disrupts the zebrafish orthologue of the X-linked human Duchenne muscular dystrophy (DMD) gene. Mutations in this locus cause Duchenne or Becker muscular dystrophies in human patients and are thought to result in a dystrophic pathology through disconnecting the cytoskeleton from the extracellular matrix in skeletal muscle by reducing the level of dystrophin protein at the sarcolemma. This is thought to allow tearing of this membrane, which in turn leads to cell death. Surprisingly, we have found that the progressive muscle degeneration phenotype of sapje mutant zebrafish embryos is caused by the failure of embryonic muscle end attachments. Although a role for dystrophin in maintaining vertebrate myotendinous junctions (MTJs) has been postulated previously and MTJ structural abnormalities have been identified in the Dystrophin-deficient mdx mouse model, in vivo evidence of pathology based on muscle attachment failure has thus far been lacking. This zebrafish mutation may therefore provide a model for a novel pathological mechanism of Duchenne muscular dystrophy and other muscle diseases

Metric preheating and limitations of linearized gravity

Recently it has become clear that the resonant amplification of quantum field fluctuations at preheating must be accompanied by resonant amplification of scalar metric perturbations, since the two are united by Einstein's equations. Furthermore, this "metric preheating" enhances particle production and leads to gravitational rescattering effects even at linear order. In multi-field models with strong preheating (q \gg 1), metric perturbations are driven nonlinear, with the strongest amplification typically on super-Hubble scales (k \to 0). This amplification is causal, being due to the super- Hubble coherence of the inflaton condensate, and is accompanied by resonant growth of entropy perturbations. The amplification invalidates the use of the linearized Einstein field equations, irrespective of the amount of fine-tuning of the initial conditions. This has serious implications at all scales - from the large-angle cosmic microwave background (CMB) anisotropies to primordial black holes. We investigate the (q,k) parameter space in a two-field model, and introduce the time to nonlinearity, t_{nl}, as the timescale for the breakdown of the linearized Einstein equations. Backreaction effects are expected to shut down the linear resonances, but cannot remove the existing amplification, which threatens the viability of strong preheating when confronted with the CMB. We discuss ways to escape the above conclusions, including secondary phases of inflation and preheating solely to fermions. Finally we rank known classes of inflation from strongest (chaotic and strongly coupled hybrid inflation) to weakest (hidden sector, warm inflation) in terms of the distortion of the primordial spectrum due to these resonances in preheating.Comment: 31 pages, 16 figures, Revtex. Final version. Nuclear Physics B (in press

Genome-wide survey of cytochrome P450 genes in the salmon louse Lepeophtheirus salmonis (Krøyer, 1837)

Background The salmon louse (Lepeophtheirus salmonis) infests farmed and wild salmonid fishes, causing considerable economic damage to the salmon farming industry. Infestations of farmed salmon are controlled using a combination of non-medicinal approaches and veterinary drug treatments. While L. salmonis has developed resistance to most available salmon delousing agents, relatively little is known about the molecular mechanisms involved. Members of the cytochrome P450 (CYP) superfamily are typically monooxygenases, some of which are involved in the biosynthesis and metabolism of endogenous compounds, while others have central roles in the detoxification of xenobiotics. In terrestrial arthropods, insecticide resistance can be based on the enhanced expression of CYPs. The reported research aimed to characterise the CYP superfamily in L. salmonis and assess its potential roles in drug resistance. Methods Lepeophtheirus salmonis CYPs were identified by homology searches of the genome and transcriptome of the parasite. CYP transcript abundance in drug susceptible and multi-resistant L. salmonis was assessed by quantitative reverse transcription PCR, taking into account both constitutive expression and expression in parasites exposed to sublethal levels of salmon delousing agents, ecdysteroids and environmental chemicals. Results The above strategy led to the identification of 25 CYP genes/pseudogenes in L. salmonis, making its CYP superfamily the most compact characterised for any arthropod to date. Lepeophtheirus salmonis possesses homologues of a number of arthropod CYP genes with roles in ecdysteroid metabolism, such as the fruit fly genes disembodied, shadow, shade, spook and Cyp18a1. CYP transcript expression did not differ between one drug susceptible and one multi-resistant strain of L. salmonis. Exposure of L. salmonis to emamectin benzoate or deltamethrin caused the transcriptional upregulation of certain CYPs. In contrast, neither ecdysteroid nor benzo[a]pyrene exposure affected CYP transcription significantly. Conclusions The parasite L. salmonis is demonstrated to possess the most compact CYP superfamily characterised for any arthropod to date. The complement of CYP genes in L. salmonis includes conserved CYP genes involved in ecdysteroid biosynthesis and metabolism, as well as drug-inducible CYP genes. The present study does not provide evidence for a role of CYP genes in the decreased susceptibility of the multiresistant parasite strain studied

Elucidating the individual effects of calcium and phosphate ions on hMSCs by using composite materials

Prova tipográficaThe biological performance of bone graft substitutes based on calcium phosphate bioceramics is dependent on a number of properties including chemical composition, porosity and surface micro- and nanoscale structure. However, in contemporary bioceramics these properties are interlinked, therefore making it difficult to investigate the individual effects of each property on cell behavior. In this study we have attempted to investigate the effects of calcium and inorganic phosphate ions independent from one another by preparing composite materials with polylactic acid (PLA) as a polymeric matrix and calcium carbonate or sodium phosphate salts as fillers. Clinically relevant bone marrow derived human mesenchymal stromal cells (hMSCs) were cultured on these composites and proliferation, osteogenic differentiation and ECM mineralization were investigated with time and were compared to plain PLA control particles. In parallel, cells were also cultured on conventional cell culture plates in media supplemented with calcium or inorganic phosphate to study the effect of these ions independent of the 3D environment created by the particles. Calcium was shown to increase proliferation of cells, whereas both calcium and phosphate positively affected alkaline phosphatase enzyme production. QPCR analysis revealed positive effects of calcium and of inorganic phosphate on the expression of osteogenic markers, in particular bone morphogenetic protein-2 and osteopontin. Higher levels of mineralization were also observed upon exposure to either ion. Effects were similar for cells cultured on composite materials and those cultured in supplemented media, although ion concentrations in the composite cultures were lower. The approach presented here may be a valuable tool for studying the individual effects of a variety of soluble compounds, including bioinorganics, without interference from other material properties.TeRM Smart Mix Program of the Netherlands Ministry of Education, Culture and Science Netherlands Science Organisation TA-COAST Grant # 05321104 Portuguese Foundation for Science and Technology (FCT) Grant # SFRH/BD69962/201

Key role of mitochondrial mutation Leu107Ser (COX1) in deltamethrin resistance in salmon lice (Lepeophtheirus salmonis)

The pyrethroid deltamethrin (DTM) is used to treat Atlantic salmon (Salmo salar) against salmon louse (Lepeophtheirus salmonis) infestations. However, DTM resistance has evolved in L. salmonis and is currently common in the North Atlantic. This study aimed to re-assess the association between DTM resistance and mitochondrial (mtDNA) mutations demonstrated in previous reports. Among 218 L. salmonis collected in Scotland in 2018–2019, 89.4% showed DTM resistance in bioassays, while 93.6% expressed at least one of four mtDNA single nucleotide polymorphisms (SNPs) previously shown to be resistance associated. Genotyping at further 14 SNP loci allowed to define three resistance-associated mtDNA haplotypes, named 2, 3 and 4, occurring in 72.0%, 14.2% and 7.3% of samples, respectively. L. salmonis strains IoA-02 (haplotype 2) and IoA-10 (haplotype 3) both showed high levels (~ 100-fold) of DTM resistance, which was inherited maternally in crossing experiments. MtDNA haplotypes 2 and 3 differed in genotype for 17 of 18 studied SNPs, but shared one mutation that causes an amino acid change (Leu107Ser) in the cytochrome c oxidase subunit 1 (COX1) and was present in all DTM resistant while lacking in all susceptible parasites. We conclude that Leu107Ser (COX1) is a main genetic determinant of DTM resistance in L. salmonis

Time-to-response toxicity analysis as a method for drug susceptibility assessment in salmon lice

The salmon louse Lepeophtheirus salmonis (Krøyer, 1837) is an ectoparasite causing infections ofwild and farmed Atlantic salmon (Salmo salar L.) in the Northern hemisphere.While L. salmonis control at commercial mariculture sites increasingly employs non-medicinal approaches, such as cage designs reducing infection rates and biological control through cleaner fish, anti-parasitic drugs are still a requirement for effective fish health care. With only a limited range of salmon delousing agents available, all of which have been in use for more than a decade, drug resistance formation has been reported for different products. Successful resistance management requires reliable susceptibility assessment, which is usually achieved through L. salmonis bioassays. These tests involve the exposure of parasites to different drug concentrations and require significant numbers of suitable L. salmonis stages. The present study reports an alternative bioassay that is based on time-to-response toxicity analyses and can be carried outwith limited parasite numbers. The assay determines the median effective time (ET50), i.e., the time required until impaired swimming and/or attachment behaviour becomes apparent in 50% of parasites, by conducting repeated examinations of test animals starting at the timepointwhere exposure to a set drug concentration commences. This experimental approach further allows the estimation of the apparent drug susceptibility of individual L. salmonis by determining their time to response, which may prove useful in experiments designed to elucidate associations between genetic factors and the drug susceptibility phenotype of parasites. Three laboratory strains of L. salmonis differing in susceptibility to emamectin benzoate were characterised using standard 24 h bioassays and time-to-response toxicity assays. While both the median effective concentration (EC50) and the ET50 showed variability between experimental repeats, both types of bioassay consistently discriminated susceptible and drug-resistant L. salmonis laboratory strains. Statement of relevance: Infections by sea lice cause significant costs to the global salmon farming industry, which have been estimated to exceed €300 million per year worldwide. Control of sea lice still relies to a significant extent on chemical delousing; however, chemical control is threatened by resistance formation. Resistance can be combated by rotation between different drugs and strategic implementation of non-medicinal strategies. However, resistance management requires reliable and feasible methods of susceptibility assessment. The present study is a technical note introducing a novel approach to susceptibility assessments in sea lice. The method can be applied in susceptibility assessments on farms,where it offers the advantage of a reduced requirement of parasites for testing. In addition, the novel method allows deriving the times of parasite require to showa response after drug treatment has started, thus providing a variable characterizing the drug susceptibility phenotype of individual parasites. Accordingly, the bioassay approach presented here will be useful for studies aiming at unravelling the genetic determinants of drug resistance

Non-Gaussian perturbations from multi-field inflation

We show how the primordial bispectrum of density perturbations from inflation may be characterised in terms of manifestly gauge-invariant cosmological perturbations at second order. The primordial metric perturbation, zeta, describing the perturbed expansion of uniform-density hypersurfaces on large scales is related to scalar field perturbations on unperturbed (spatially-flat) hypersurfaces at first- and second-order. The bispectrum of the metric perturbation is thus composed of (i) a local contribution due to the second-order gauge-transformation, and (ii) the instrinsic bispectrum of the field perturbations on spatially flat hypersurfaces. We generalise previous results to allow for scale-dependence of the scalar field power spectra and correlations that can develop between fields on super-Hubble scales.Comment: 11 pages, RevTex; minor changes to text; conclusions unchanged; version to appear in JCA

Mutations in voltage-gated sodium channels from pyrethroid resistant salmon lice (Lepeophtheirus salmonis)

Background Parasitic salmon lice (Lepeophtheirus salmonis) cause high economic losses in Atlantic salmon farming. Pyrethroids, which block arthropod voltage‐gated sodium channels (Nav1), are used for salmon delousing. However, pyrethroid resistance is common in L. salmonis. The present study characterised Nav1 homologues in L. salmonis in order to identify channel mutations associated to resistance, called kdr (knockdown) mutations. Results Genome scans identified three L. salmonis Nav1 homologues, LsNav1.1, LsNav1.2 and LsNav1.3. Arthropod kdr mutations map to specific Nav1 regions within domains DI‐III, namely segments S5 and S6 and the linker helix connecting S4 and S5. The above channel regions were amplified by RT‐PCR and sequenced in deltamethrin‐susceptible and deltamethrin‐resistant L. salmonis. While LsNav1.1 and LsNav1.2 lacked nucleotide polymorphisms showing association to resistance, LsNav1.3 showed a non‐synonymous mutation in S5 of DII occurring in deltamethrin‐resistant parasites. The mutation is homologous to a previously described kdr mutation (I936V, numbering according to Musca domestica Vssc1) and was present in two pyrethroid‐resistant L. salmonis strains (allele frequencies of 0.800 and 0.357), but absent in two pyrethroid‐susceptible strains. Conclusions The present study indicates that a kdr‐mutation in LsNaV 1.3 may contribute to deltamethrin resistance in L. salmonis