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An integrated approach that combines geophysical surveys and numerical simulations is proposed to study the processes that govern the fluid flow along active fault zones. It is based on the reconstruction of the architecture of the investigated fault system, as well as the identification of possible paths for fluid migration, according to the distribution of geophysical parameters retrieved by multi-methodological geophysical prospecting. The aim is to establish, thanks to constraints deriving from different types of data (e.g., geological, geochemical and/or hydrogeological data), an accurate 3D petrophysical model of the survey area to be used for simulating, by numerical modelling, the physical processes likely taking place in the imaged system and its temporal evolution. The effectiveness of the proposed approach is tested in an active fault zone in the Matese Mts (southern Italy), where recent, accurate geochemical measurements have registered very high anomalous values of non-volcanic natural emissions of CO2. In particular, a multi-methodological geophysical survey, consisting of electrical resistivity tomography, self-potential and passive seismic measurements, integrated with geological data, was chosen to define the 3D petrophysical model of the investigated system and to identify possible source geometries. Three different scenarios were assumed corresponding to three different CO2 source models. The one that hypothesizes a source located along the fault plane at the depth of the carbonate basement was found to be the best candidate to represent the test site. Indeed, the performed numerical simulations provide CO2 flow estimates comparable with the values observed in the investigated area. These findings are promising for gas hazards, as they suggest that numerical simulations of different CO2 degassing scenarios could forecast possible critical variations in the amount of CO2 emitted near the fault. [ABSTRACT FROM AUTHOR]

Gas leakage from deep geologic storage formations to the Earth's surface is one of the main hazards in geological carbon sequestration and storage. Permeable sediment covers together with natural pathways, such as faults and/or fracture systems, are the main factors controlling surface leakages. Therefore, the characterization of natural systems, where large amounts of natural gases are released, can be helpful for understanding the effects of potential gas leaks from carbon dioxide storage systems. In this framework, we propose a combined use of high-resolution geoelectrical investigations (i.e. resistivity tomography and self-potential surveys) for reconstructing shallow buried fracture networks in the caprock and detecting preferential gas migration pathways before it enters the atmosphere. Such methodologies appear to be among the most suitable for the research purposes because of the strong dependence of the electrical properties of water-bearing permeable rock, or unconsolidated materials, on many factors relevant to CO 2 storage (i.e. porosity, fracturing, water saturation, etc.). The effectiveness of the suggested geoelectrical approach is tested in an area of natural gas degassing (mainly CH 4 ) located in the active fault zone of the Bolle della Malvizza (Southern Apennines, Italy), which could represent a natural analogue of gas storage sites due to the significant thicknesses (hundreds of meters) of impermeable rock (caprock) that is generally required to prevent carbon dioxide stored at depth from rising to the surface. The obtained 3D geophysical model, validated by the good correlation with geochemical data acquired in the study area and the available geological information, provided a structural and physical characterization of the investigated subsurface volume. Moreover, the time variations of the observed geophysical parameters allowed the identification of possible migration pathways of fluids to the surface., (© 2023. The Author(s).)

SQUAMOSA PROMOTER BINDING-LIKE (SPL) proteins constitute a large family of transcription factors known to play key roles in growth and developmental processes, including juvenile-to-adult and vegetative-to-reproductive phase transitions. This makes SPLs interesting targets for precision breeding in plants of the Nicotiana genus used as e.g. recombinant biofactories. We report the identification of 49 SPL genes in Nicotiana tabacum cv. K326 and 43 SPL genes in Nicotiana benthamiana LAB strain, which were classified into eight phylogenetic groups according to the SPL classification in Arabidopsis. Exon-intron gene structure and DNA-binding domains were highly conserved between homeologues and orthologues. Thirty of the NbSPL genes and 33 of the NtSPL genes were found to be possible targets of microRNA 156. The expression of SPL genes in leaves was analysed by RNA-seq at three different stages, revealing that genes not under miR156 control were in general constitutively expressed at high levels, whereas miR156-regulated genes showed lower expression, often developmentally regulated. We selected the N. benthamiana SPL13_1a gene as target for a CRISPR/Cas9 knock-out experiment. We show here that a full knock-out in this single gene leads to a significant delay in flowering time, a trait that could be exploited to increase biomass for recombinant protein production., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Objective: Migration is a phenomenon related to adverse impacts, including higher risk for psychological distress and trauma symptoms, highlighting the strong need for effective psychological treatments to help migrants. However, the use of expressive writing intervention (EW) has not been examined in this population. The aims of the study were to evaluate: (a) the effectiveness of a trauma-focused EW on psychological distress, trauma symptoms, alexithymia, and hope for the future in migrants, and (b) the role of alexithymia and hope for the future in the association with the EW and the psychological symptoms., Method: Twenty-eight migrants were enrolled in a multiarm double-blind randomized controlled trial (RCT) that included three groups: (a) trauma-focused EW, (b) neutral EW, and (c) control. Participants completed self-report measures at pretreatment, immediately after the intervention, and at 1 month follow-up. The study complied with the guidelines of Consolidate Standard of Reporting Trials (CONSORT) checklist and was retrospectively recorded on ClinicalTrials.gov., Results: Trauma-focused EW was related to an immediate improvement in phobic anxiety and positive total symptoms and improvement in somatization, global severity index, and hope for the future at follow-up. Trauma symptoms and alexithymia did not show significant effects. Factorial regressions showed that the interaction group per time per hope for the future was a significant predictor on the phobic anxiety., Discussion: EW may be an effective tool for reducing migrant's distress and may have long-term improvements in mental health. Moreover, the results suggest the potential influence of the hope for the future on anxiety in migrants. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

CRISPR/Cas ability to target several loci simultaneously (multiplexing) is a game-changer in plant breeding. Multiplexing not only accelerates trait pyramiding but also can unveil traits hidden by functional redundancy. Furthermore, multiplexing enhances dCas-based programmable gene expression and enables cascade-like gene regulation. However, the design and assembly of multiplex constructs comprising tandemly arrayed guide RNAs (gRNAs) requires scarless cloning and is still troublesome due to the presence of repetitive sequences, thus hampering a more widespread use. Here we present a comprehensive extension of the software-assisted cloning platform GoldenBraid (GB), in which, on top of its multigene cloning software, we integrate new tools for the Type IIS-based easy and rapid assembly of up to six tandemly-arrayed gRNAs with both Cas9 and Cas12a, using the gRNA-tRNA-spaced and the crRNA unspaced approaches, respectively. As stress tests for the new tools, we assembled and used for Agrobacterium-mediated stable transformation a 17 Cas9-gRNAs construct targeting a subset of the Squamosa-Promoter Binding Protein-Like (SPL) gene family in Nicotiana tabacum . The 14 selected genes are targets of miR156, thus potentially playing an important role in juvenile-to-adult and vegetative-to-reproductive phase transitions. With the 17 gRNAs construct we generated a collection of Cas9-free SPL edited T 1 plants harboring up to 9 biallelic mutations and showing leaf juvenility and more branching. The functionality of GB-assembled dCas9 and dCas12a-based CRISPR/Cas activators and repressors using single and multiplexing gRNAs was validated using a Luciferase reporter with the Solanum lycopersicum Mtb promoter or the Agrobacterium tumefaciens nopaline synthase promoter in transient expression in Nicotiana benthamiana . With the incorporation of the new web-based tools and the accompanying collection of DNA parts, the GB4.0 genome edition turns an all-in-one open platform for plant genome engineering., Competing Interests: JQ was employed by the company Idoasis 2002 S.L. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Vazquez-Vilar, Garcia-Carpintero, Selma, Bernabé-Orts, Sanchez-Vicente, Salazar-Sarasua, Ressa, de Paola, Ajenjo, Quintela, Fernández-del-Carmen, Granell and Orzáez.)

The Confederation of Postgraduate Medical Education Councils launched the Australian Curriculum Framework for Junior Doctors in October 2006. The curriculum framework: balances the major areas of clinical management, communication and professionalism, and highlights the importance of an integrated approach to prevocational learning and teaching; supports practice-based, opportunistic and continuous learning, and specifies performance and supervision requirements for junior doctors; and has been published in both Internet and printable versions, to make the document accessible and easily usable by junior doctors and supervisors. The implementation of the curriculum framework will be overseen by a steering group that includes representatives from key stakeholder groups, including junior doctors and medical students.

Hydrolases containing two metal ions connected by a bridging ligand catalyze reactions important in carcinogensis, tissue repair, post-translational modification, control and regulation of biochemical pathways, and protein degradation. The aminopeptidase from Aeromonas proteolytica serves as a paradigm for the study of such bridged bimetallic proteases since its three-dimensional structure is known to very high resolution and its catalytic reaction is amenable to spectroscopic examination. Herein, we report the X-ray crystal structure at 1.9 A resolution of AAP complexed with 1-butaneboronic acid (BuBA). This structure suggests that this complex represents a snapshot of the proteolytic reaction in an arrested form between the Michaelis complex and the transition state. Comparison of the structure with spectroscopic and other data allows us to conclude that the apparently structurally symmetrical dizinc site is actually asymmetric electrostatically.

Objective: Epidemiologic evidence of a role for antioxidants in the prevention of chronic disease has been inconclusive, in part due to the difficulty of measuring past diets of free-living populations. The purpose of the current study was to examine the reliability of a 19-item, self-administered, semiquantitative, food frequency questionnaire to assess intake of the major dietary antioxidants., Methods: Reliability was established by administering the food frequency questionnaire a second time by telephone. The subjects comprised 151 participants in the Melbourne Visual Impairment Project, a study of the distribution and determinants of eye disease in Melbourne residents aged 40 and over., Results: Spearman correlation coefficients ranged from 0.39 for spinach to 0.76 for yoghurt, and all were highly significant (all p = 0.001). The reliability of the instrument was not influenced by gender, English speaking ability, or the number of days between the first and second administration of the questionnaire., Conclusion: In conclusion, we have shown this 19-item food frequency questionnaire to be highly reliable. It should be useful for anyone involved in the study of the relationship of dietary antioxidant intake to health outcomes in large populations where limitations of time and money prohibit the collection of more detailed dietary intake information.

Objective: To determine the level of knowledge of glaucoma in a population-based sample, and its relationship to self-care practices., Design and Subjects: A cluster random sample of the Melbourne population 40 years of age and older was interviewed. One thousand seven hundred and eleven residents living in five randomly selected Melbourne metropolitan suburbs, each consisting of two adjacent census collector districts., Measures: Questions were asked concerning respondents' awareness, knowledge and description of the disease. Respondents were also asked the year of their last visit to their eye health care provider., Results: Seventy per cent of the sample had heard of glaucoma. However, only 22% provided a description that demonstrated a reasonable understanding of the disease. A lack of awareness and knowledge of glaucoma appeared to be negatively related to self-care practices., Conclusion: Serious deficiencies in the basic knowledge of glaucoma in the community was demonstrated. This has significant public health implications as only a small percentage of the at-risk population may present themselves for assessment and treatment. Informing the community about glaucoma is an important step in promoting preventative ophthalmic care and reducing visual impairment and blindness.

The synthesis of esters derived from 7-(diphenylmethylene)bicyclo[2.2.1]heptan-2-endo-ol, obtained by LiAlH4 reduction of 7-(diphenylmethylene)bicyclo[2.2.1]heptan-2-one, is described. Some of these esters showed an appreciable antiarrhythmic activity in rats, as well as moderate hypotensive and local anesthetic activities in rats and mice, respectively.

The syntheses of 1-(2-hydroxy-2-phenylethyl)-3,5-diphenyl-1H-pyrazole 1 by reaction of 2-hydrazino-1-phenylethanol with dibenzoylmethane, of esters 2 and 2-dialkylaminoethyl ethers 3 starting from 1 as sodium salt and acyl chlorides or 2-chloroethyldialkylamines, respectively, as well as of N,N-disubstituted 1-(2-amino-2-phenylethyl)-3,5-diphenyl-1H-pyrazoles 5 by reaction of secondary amines with the tosylate of 1, are described. Some of the above compounds showed a considerable sedative effect in mice and a remarkable platelet antiaggregating activity in vitro, as well as moderate local anesthetic, analgesic and antiinflammatory activities in mice and rats.

The study has been carried out on albino rats pre and/or postnatally exposed to vanadate (1 and 10 mg/dl). In pups pre- and postnatal or postnatal vanadate exposure significantly decreased on the 30th and 60th day of age the pressor response to 1-noradrenaline, and increased the pressor response to sinus-carotid baroreceptor stimulation and the hypotensive responses to 1-isoprenaline and acetylcholine. The effects were observed at vanadate doses that did not interfere with gestation, maternal and pup body weight, maternal systolic arterial blood pressure during pregnancy, and pup systolic arterial blood pressure. These doses have not determined any macroscopic teratogenic effects.

Experimental research has documented that fenoverine is a depressant drug mostly active on the intestinal smooth muscle; however, it is also active on other tissues as well as on the genito-urinary tract. Fenoverine may be considered a synchronizer of smooth muscle effective in modulating the intracellular influx of calcium into the cell and its release from the intracellular pool. Fenoverine causes a reduction of the excitatory junction potential (EJP) in intestinal smooth muscle by stimulating parasympathetic efferent fibres without any change in the inhibitory junction potential (IJP). This also occurs in the presence of atropine. Fenoverine reduces the intracellular Ca2+ concentration by decreasing the calcium gradient across the cell membrane, as well as decreasing the release of Ca2+ from the intestinal pool more intensely than papaverine (but less intensely than nifedipine, verapamil or diltiazem). This phenomenon has been observed in the intestinal smooth muscle and also in the genito-urinary tract.