Targeting the autosomal Ceratitis capitata transformer gene using Cas9 or dCas9 to masculinize XX individuals without inducing mutations

Background: Females of the Mediterranean fruit fly Ceratitis capitata (Medfly) are major agricultural pests, as they lay eggs into the fruit crops of hundreds of plant species. In Medfly, female sex determination is based on the activation of Cctransformer (Cctra). A maternal contribution of Cctra is required to activate Cctra itself in the XX embryos and to start and epigenetically maintain a Cctra positive feedback loop, by female-specific alternative splicing, leading to female development. In XY embryos, the male determining Maleness-on-the-Y gene (MoY) blocks this activation and Cctra produces male-specific transcripts encoding truncated CcTRA isoforms and male differentiation occurs. Results: With the aim of inducing frameshift mutations in the first coding exon to disrupt both female-specific and shorter male-specific CcTRA open reading frames (ORF), we injected Cas9 ribonucleoproteins (Cas9 and single guide RNA, sgRNA) in embryos. As this approach leads to mostly monoallelic mutations, masculinization was expected only in G1 XX individuals carrying biallelic mutations, following crosses of G0 injected individuals. Surprisingly, these injections into XX-only embryos led to G0 adults that included not only XX females but also 50% of reverted fertile XX males. The G0 XX males expressed male-specific Cctra transcripts, suggesting full masculinization. Interestingly, out of six G0 XX males, four displayed the Cctra wild type sequence. This finding suggests that masculinization by Cas9-sgRNA injections was independent from its mutagenic activity. In line with this observation, embryonic targeting of Cctra in XX embryos by a dead Cas9 (enzymatically inactive, dCas9) also favoured a male-specific splicing of Cctra, in both embryos and adults. Conclusions: Our data suggest that the establishment of Cctra female-specific autoregulation during the early embryogenesis has been repressed in XX embryos by the transient binding of the Cas9-sgRNA on the first exon of the Cctra gene. This hypothesis is supported by the observation that the shift of Cctra splicing from female to male mode is induced also by dCas9. Collectively, the present findings corroborate the idea that a transient embryonic inactivation of Cctra is sufficient for male sex determination

Il riequilibrio della finanza pubblica negli anni Novanta

Gli anni novanta sono caratterizzati dal riequilibrio della finanza pubblica: l’indebitamento netto delle Amministrazioni pubbliche che nel 1990 era pari all’11,8 per cento del PIL scende nel 1999 all’1,8 per cento. Gli anni ottanta possono essere giudicati ex post un periodo nel quale è tardato il riconoscimento della necessità del rigore fiscale, con un indebitamento netto mediamente pari al 10,8 del PIL e pressoché costante nel periodo. Questa caratterizzazione valevole per l’intero decennio, può essere meglio precisata. Ad esempio, l’anno 1991 (e non solo il 1990) può essere collegato a quelli del quinquennio precedente, che è stato definito, giustamente, come quello del “risanamento mancato” ; il 1992 è l’anno della “svolta” ; il 1997 l’anno in cui il riequilibrio, con gran prontezza, è stato colto con “astuzia e virtù”. Tuttavia, secondo gli autori di questa nota, sarebbe erronea una suddivisione del decennio in alcuni episodi di manovre di finanza pubblica, ora fallite ora riuscite, come alcune volte sono stati presentati nella letteratura; il processo di riequilibrio è stato un percorso continuo, ancor più di quanto appaia nei dati grezzi dell’indebitamento netto e dell’avanzo primario, sia nella programmazione e nella attuazione delle manovre fiscali, sia per la continuità dei risultati. Ovviamente alcuni momenti del processo sono stati molto qualificanti, e saranno posti in rilievo nell’analisi. Il successo della politica fiscale perseguita nel decennio, non esime comunque dal porsi e risolvere ulteriori problemi, derivanti nel medio periodo dalle pressioni demografiche, da possibili riforme istituzionali e dall’intensificarsi della competitività tra i vari sistemi - paese

Monge Distance between Quantum States

We define a metric in the space of quantum states taking the Monge distance between corresponding Husimi distributions (Q--functions). This quantity fulfills the axioms of a metric and satisfies the following semiclassical property: the distance between two coherent states is equal to the Euclidean distance between corresponding points in the classical phase space. We compute analytically distances between certain states (coherent, squeezed, Fock and thermal) and discuss a scheme for numerical computation of Monge distance for two arbitrary quantum states.Comment: 9 pages in LaTex - RevTex + 2 figures in ps. submitted to Phys. Rev.

CPO and quantitative textural analyses within sheath folds

Acknowledgments This has been a multi-national collaboration from authors based in Europe, North America, Australia and India. Erasmus funding to GIA in 2018 enabled a visit to Catania leading to discussion and initiation of this project. The authors are grateful to Amarnath Dandapat for preparation of superpolished rock thin sections at the Department of Geology and Geophysics (IIT Kharagpur, India). Niloy Bhowmik is thanked for assistance with SEM-EBSD data generation in the Central Research Facility (IIT Kharagpur, India). E.F. thanks Sibio Carmelo for thin sections preparation at the University of Turin (Italy). Authors are grateful to ANSTO laboratory personnel for the preparation of specimens (funded proposals: P9835 with the title “Sheath fold texture characterisation”, principal scientist: E.F.; co-proposers: G.I.A. and V.L.; DB6749 with the title “Texture analysis of rocks”, principal scientist: V.L.; co-proposer: E.F.; DB9606 with the title “A pilot experiment for texture characterisation in a sheath fold”, principal scientist: E.F..; co-proposers: G.I.A. and V.L.). L.N. and R.G. report that this publication has been assigned the NRCan contribution number 20230109. Many thanks to Richard D. Law and an anonymous reviewer for their careful revision that substantially improved the original version of the manuscript. We also thanks Dr. T.K. Cawood from the Geological Survey of Canada for her useful comments on the drafted manuscript. The editorial handling by Fabrizio Agosta is greatly appreciated.Peer reviewedPublisher PD

Positive selection in Europeans and East-Asians at the ABCA12 gene

Natural selection acts on genetic variants by increasing the frequency of alleles responsible for a cellular function that is favorable in a certain environment. In a previous genome-wide scan for positive selection in contemporary humans, we identified a signal of positive selection in European and Asians at the genetic variant rs10180970. The variant is located in the second intron of the ABCA12 gene, which is implicated in the lipid barrier formation and down-regulated by UVB radiation. We studied the signal of selection in the genomic region surrounding rs10180970 in a larger dataset that includes DNA sequences from ancient samples. We also investigated the functional consequences of gene expression of the alleles of rs10180970 and another genetic variant in its proximity in healthy volunteers exposed to similar UV radiation. We confirmed the selection signal and refine its location that extends over 35 kb and includes the first intron, the first two exons and the transcription starting site of ABCA12. We found no obvious effect of rs10180970 alleles on ABCA12 gene expression. We reconstructed the trajectory of the T allele over the last 80,000 years to discover that it was specific to H. sapiens and present in non-Africans 45,000 years ago

Integrating Human Factors Engineering and Information Processing Approaches to Facilitate Evaluations in Criminal Justice Technology Research

Background: Evaluations are routinely conducted by government agencies and research organizations to assess the effectiveness of technology in criminal justice. Interdisciplinary research methods are salient to this effort. Technology evaluations are faced with a number of challenges including (1) the need to facilitate effective communication between social science researchers, technology specialists, and practitioners, (2) the need to better understand procedural and contextual aspects of a given technology, and (3) the need to generate findings that can be readily used for decision making and policy recommendations. Objectives: Process and outcome evaluations of technology can be enhanced by integrating concepts from human factors engineering and information processing. This systemic approach, which focuses on the interaction between humans, technology, and information, enables researchers to better assess how a given technology is used in practice. Subjects: Examples are drawn from complex technologies currently deployed within the criminal justice system where traditional evaluations have primarily focused on outcome metrics. Although this evidence-based approach has significant value, it is vulnerable to fully account for human and structural complexities that compose technology operations. Conclusions: Guiding principles for technology evaluations are described for identifying and defining key study metrics, facilitating communication within an interdisciplinary research team, and for understanding the interaction between users, technology, and information. The approach posited here can also enable researchers to better assess factors that may facilitate or degrade the operational impact of the technology and answer fundamental questions concerning whether the technology works as intended, at what level, and cost

A Serum Resistin and Multicytokine Inflammatory Pathway Is Linked With and Helps Predict All-cause Death in Diabetes

Context: Type 2 diabetes (T2D) shows a high mortality rate, partly mediated by atherosclerotic plaque instability. Discovering novel biomarkers may help identify high-risk patients who would benefit from more aggressive and specific managements. We recently described a serum resistin and multicytokine inflammatory pathway (REMAP), including resistin, interleukin (IL)-1 beta, IL-6, IL-8, and TNF-alpha, that is associated with cardiovascular disease.Objective: We investigated whether REMAP is associated with and improves the prediction of mortality in T2D.Methods: A REMAP score was investigated in 3 cohorts comprising 1528 patients with T2D (409 incident deaths) and in 59 patients who underwent carotid endarterectomy (CEA; 24 deaths). Plaques were classified as unstable/stable according to the modified American Heart Association atherosclerosis classification.Results: REMAP was associated with all-cause mortality in each cohort and in all 1528 individuals (fully adjusted hazard ratio [HR] for 1 SD increase=1.34, P<.001). In CEA patients, REMAP was associated with mortality (HR=1.64, P=.04) and a modest change was observed when plaque stability was taken into account (HR=1.58; P=.07). REMAP improved discrimination and reclassification measures of both Estimation of Mortality Risk in Type 2 Diabetic Patients and Risk Equations for Complications of Type 2 Diabetes, well-established prediction models of mortality in T2D (P<.05-<.001).Conclusion: REMAP is independently associated with and improves predict all-cause mortality in T2D; it can therefore be used to identify high-risk individuals to be targeted with more aggressive management. Whether REMAP can also identify patients who are more responsive to IL-6 and IL-1 beta monoclonal antibodies that reduce cardiovascular burden and total mortality is an intriguing possibility to be tested

Aedes albopictus bionomics data collection by citizen participation on procida island, a promising mediterranean site for the assessment of innovative and community-based integrated pest management methods

In the last decades, the colonization of Mediterranean Europe and of other temperate regions by Aedes albopictus created an unprecedented nuisance problem in highly infested areas and new public health threats due to the vector competence of the species. The Sterile Insect Technique (SIT) and the Incompatible Insect Technique (IIT) are insecticide-free mosquito-control methods, relying on mass release of irradiated/manipulated males, able to complement existing and only partially effective control tools. The validation of these approaches in the field requires appropriate experimental settings, possibly isolated to avoid mosquito immigration from other infested areas, and preliminary ecological and entomological data. We carried out a 4-year study in the island of Procida (Gulf of Naples, Italy) in strict collaboration with local administrators and citizens to estimate the temporal dynamics, spatial distribution, and population size of Ae. albopictus and the dispersal and survival of irradiated males. We applied ovitrap monitoring, geo-spatial analyses, mark-release-recapture technique, and a citizen-science approach. Results allow to predict the seasonal (from April to October, with peaks of 928-9,757 males/ha) and spatial distribution of the species, highlighting the capacity of Ae. albopictus population of Procida to colonize and maintain high frequencies in urban as well as in sylvatic inhabited environments. Irradiated males shown limited ability to disperse (mean daily distance travelled <60m) and daily survival estimates ranging between 0.80 and 0.95. Overall, the ecological characteristics of the island, the acquired knowledge on Ae. albopictus spatial and temporal distribution, the high human and Ae. albopictus densities and the positive attitude of the resident population in being active parts in innovative mosquito control projects provide the ground for evidence-based planning of the interventions and for the assessment of their effectiveness. In addition, the results highlight the value of creating synergies between research groups, local administrators, and citizens for affordable monitoring (and, in the future, control) of mosquito populations