Spatial genetic structure in the clonal marine angiosperm <i>Cymodocea nodosa</i>: the influence of dispersal potential, mating system and species interactions

In the present thesis, the factors influencing population’s genetic structure in the clonal marine angiosperm Cymodocea nodosa have been investigated. C. nodosa is a dioecious seagrass, exhibiting both vegetative propagation and sexual reproduction. Seed dispersal is expected to be extremely restricted. I selected seven microsatellite loci through genomic library screening to investigate the relative effects of sexual and clonal reproduction on the genetic diversity and structure in a Cymodocea nodosa population from the Gulf of Naples (Italy). High clonal diversity and genet density were found. Autocorrelation analyses confirmed the expectations of very restricted seed dispersal (observed dispersal range 1-21m) in this species. The effect of mating system on genetic structure were investigated comparing the clonal architectures of the dioecious Cymodocea nodosa and monoecious Zostera noltii. An intermingled configuration of genets has been found in the dioecious Cymodocea nodosa while a clumped distribution of clones in the hermaphroditic Zostera noltii has been observed. I hypothesise that the possibility of reduction in the seed-set would drive genet distribution. On a phylogeographic spatial scale, the existence of population differentiation and of genetic disjunction within the Mediterranean Sea was investigated in Cymodocea nodosa. Populations displayed a wide variability in clonal diversity. A Bayesian analysis revealed that “supra-population” panmictic units are present in the Mediterranean basin. Genetic substructure from a phylogeographic tree coincided with major geographical boundaries within the Mediterranean basin. In general, in Cymodocea nodosa, seed dispersal is poor at the within-population level, but long-range dispersal events can occur, allowing high levels of gene flow at a phylogeographic scale. The observed “guerrilla” clonal architecture allows to reduce the effect of genetic identity on the genetic structure of the population, but it is also advantageous by allowing pollen availability and therefore a sufficient seed-set in this dioecious species

Recent and Ancient Signature of Balancing Selection around the S-Locus in Arabidopsis halleri and A. lyrata

Balancing selection can maintain different alleles over long evolutionary times. Beyond this direct effect on the molecular targets of selection, balancing selection is also expected to increase neutral polymorphism in linked genome regions, in inverse proportion to their genetic map distances from the selected sites. The genes controlling plant self-incompatibility are subject to one of the strongest forms of balancing selection, and they show clear signatures of balancing selection. The genome region containing those genes (the S-locus) is generally described as nonrecombining, and the physical size of the region with low recombination has recently been established in a few species. However, the size of the region showing the indirect footprints of selection due to linkage to the S-locus is only roughly known. Here, we improved estimates of this region by surveying synonymous polymorphism and estimating recombination rates at 12 flanking region loci at known physical distances from the S-locus region boundary, in two closely related self-incompatible plants Arabidopsis halleri and A. lyrata. In addition to studying more loci than previous studies and using known physical distances, we simulated an explicit demographic scenario for the divergence between the two species, to evaluate the extent of the genomic region whose diversity departs significantly from neutral expectations. At the closest flanking loci, we detected signatures of both recent and ancient indirect effects of selection on the S-locus flanking genes, finding ancestral polymorphisms shared by both species, as well as an excess of derived mutations private to either species. However, these effects are detected only in a physically small region, suggesting that recombination in the flanking regions is sufficient to quickly break up linkage disequilibrium with the S-locus. Our approach may be useful for distinguishing cases of ancient versus recently evolved balancing selection in other systems

Multiparameter quantum estimation of noisy phase shifts

Phase estimation is the most investigated protocol in quantum metrology, but its performance is affected by the presence of noise, also in the form of imperfect state preparation. Here we discuss how to address this scenario by using a multiparameter approach, in which noise is associated to a parameter to be measured at the same time as the phase. We present an experiment using two-photon states, and apply our setup to investigating optical activity of fructose solutions. Finally, we illustrate the scaling laws of the attainable precisions with the number of photons in the probe state

Quantum sensors for dynamical tracking of chemical processes

Quantum photonics has demonstrated its potential for enhanced sensing. Current sources of quantum light states tailored to measuring, allow to monitor phenomena evolving on time scales of the order of the second. These are characteristic of product accumulation in chemical reactions of technologically interest, in particular those involving chiral compounds. Here we adopt a quantum multiparameter approach to investigate the dynamic process of sucrose acid hydrolysis as a test bed for such applications. The estimation is made robust by monitoring different parameters at once

Intracellular Water Lifetime as a Tumor Biomarker to Monitor Doxorubicin Treatment via FFC-Relaxometry in a Breast Cancer Model

This study aims to explore whether the water exchange rate constants in tumor cells can act as a hallmark of pathology status and a reporter of therapeutic outcomes. It has been shown, using 4T1 cell cultures and murine allografts, that an early assessment of the therapeutic effect of doxorubicin can be detected through changes in the cellular water efflux rate constant k(io.) The latter has been estimated by analyzing the magnetization recovery curve in standard NMR T(1) measurements when there is a marked difference in the proton relaxation rate constants (R(1)) between the intra- and the extra-cellular compartments. In cellular studies, T(1) measurements were carried out on a relaxometer working at 0.5 T, and the required difference in R(1) between the two compartments was achieved via the addition of a paramagnetic agent into the extracellular compartment. For in-vivo experiments, the large difference in the R(1) values of the two-compartments was achieved when the T(1) measurements were carried out at low magnetic field strengths. This task was accomplished using a Fast Field Cycling (FFC) relaxometer that was properly modified to host a mouse in its probe head. The decrease in k(io) upon the administration of doxorubicin is the result of the decreased activity of Na(+)/K(+)-ATPase, as shown in an independent test on the cellular uptake of Rb ions. The results reported herein suggest that k(io) can be considered a non-invasive, early and predictive biomarker for the identification of responsive patients immediately from the first doxorubicin treatment

In vivo assessment of tumour associated macrophages in murine melanoma obtained by low-field relaxometry in the presence of iron oxide particles

Acknowledgements The authors would like to acknowledge Dr Dana Dawson, University of Aberdeen, UK, for the supply of ferumoxytol. This project has received funding from the European Union Horizon 2020 research and innovation programme under grant agreement No 668119 (project “IDentIFY”) and it was performed in the frame of the COST Action AC15209 (EURELAX). Maria Rosaria Ruggiero was supported by a “FIRC-AIRC fellowship for Italy”. The Italian Ministry for Education and Research (MIUR) is gratefully acknowledged for yearly FOE funding to the Euro-BioImaging Multi-Modal Molecular Imaging Italian Node (MMMI). Data availability All data analysed during this study are included in this published article (and its supplementary information file). Other raw data required to reproduce these findings are available from the corresponding author on reasonable request.Peer reviewedPublisher PD

Low-Field NMR Relaxometry for Intraoperative Tumour Margin Assessment in Breast-conserving Surgery

Funding: This project has received funding from the European Union Horizon 2020 research and innovation programme under grant agreement no. 668119 (project “IDentIFY”) and from AIRC under IG 2019, ID 23267 project (PI Geninatti Crich Simonetta). The authors acknowledge the Italian Ministry of Research for FOE contribution to the Euro-BioImaging MultiModal Molecular Imaging Italian Node (www.mmmi.unito.it accessed on 1 October 2020). Maria Rosaria Ruggiero was supported by a “FIRC-AIRC fellowship for Italy”. Acknowledgments: This work was performed in the frame of the COST Action CA15209 (EURELAX).Peer reviewedPublisher PD

82. Cftr Gene Targeting in Murine ES Cells Mediated by the SFHR Technique

Small Fragment Homologous Recombination (SFHR)-mediated targeting is a gene therapy strategy where a specific genomic locus is modified through a target exchange between a small DNA fragment (SDF) and genomic DNA. Here we demonstrate that SFHR can stably introduce a 3-bp deletion (corresponding to |[Delta]|F508) within Cftr (Cystic Fibrosis Transmembrane Conductance Regulator) locus in the genome of mouse embryonic stem (ES) cells. SDFs (about 6.4|[times]|105 molecules per cell) carrying the |[Delta]|F508 mutation were transfected by nucleofection protocol. About 12% of transcript corresponding to deleted allele was detected and about 60% of the electroporated cells no longer had measurable CFTR-dependent chloride efflux. The CFTR activity was also analyzed by measuring the chloride efflux by the fluorescence microscopy-coupled digital video imaging system in each ES cell colony, previously loaded with MQAE, a chloride sensitive dye. An average of 4-6 regions for each cell colony was analysed to verify the genotypic homogeneity of each colony. In fact all regions examined in each colony showed a similar significant chloride efflux after PKA activation. Moreover on twelve electroporated ES colonies analysed, eight were successfully mutated (Cl- efflux not significantly different from zero) while four colonies showed Cl-efflux CFTR-dependent not significantly different from the untreated ones

Active elderly and health-can moderate exercise improve health and wellbeing in older adults? Protocol for a randomized controlled trial

Abstract Background: Aging is marked by a progressive rise in chronic diseases with an impact on social and healthcare costs. Physical activity (PA) may soothe the inconveniences related to chronic diseases, has positive effects on the quality of life and biological rhythms, and can prevent the decline in motor functions and the consequent falls, which are associated with early death and disability in older adults. Methods: We randomized 120 over-65 males and females into groups of similar size and timing and will give each either moderate physical activity or cultural and recreational activities. Being younger than 65 years, inability to participate in physical activity for any medical reason, and involvement in a massive program of physical exercise are the exclusion criteria. The primary outcome measures are quality of life, walking speed, and postural sway. Participants are tested at baseline, post-treatment, and 6-month (24 weeks) and 12-month (48 weeks) follow-ups. Discussion: This study aims at improving the quality of life, wellness, and cognitive functioning in the elderly through a low-cost affordable program of moderate physical activity. Given the growing aging of the world population and the social and economic burden of disability in the elderly, our results might have a major impact on future practices