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Background Different studies have proved in recent years that hypofractionated radiotherapy (RT) improves overall survival of patients affected by locally advanced, unresectable, pancreatic cancer. The clinical management of these patients generally leads to poor results and is considered very challenging, due to different factors, heavily influencing treatment delivery and its outcomes. Firstly, the dose prescribed to the target is limited by the toxicity that the highly radio-sensitive organs at risk (OARs) surrounding the disease can develop. Treatment delivery is also complicated by the significant inter-fractional and intra-fractional variability of therapy volumes, mainly related to the presence of hollow organs and to the breathing cycle. Main body of the abstract The recent introduction of magnetic resonance guided radiotherapy (MRgRT) systems leads to the opportunity to control most of the aforementioned sources of uncertainty influencing RT treatment workflow in pancreatic cancer. MRgRT offers the possibility to accurately identify radiotherapy volumes, thanks to the high soft-tissue contrast provided by the Magnetic Resonance imaging (MRI), and to monitor the tumour and OARs positions during the treatment fraction using a high-temporal cine MRI. However, the main advantage offered by the MRgRT is the possibility to online adapt the RT treatment plan, changing the dose distribution while the patient is still on couch and successfully addressing most of the sources of variability. Short conclusion Aim of this study is to present and discuss the state of the art, the main pitfalls and the innovative opportunities offered by online adaptive MRgRT in pancreatic cancer treatment.

The development of spectral sensors (SSs) capable of retrieving spectral information have opened new opportunities to improve several environmental and agricultural practices, e.g., crop breeding, plant phenotyping, land use monitoring, and crop classification. The SSs are classified as multispectral and hyperspectral (HS) based on the number of the spectral bands resolved and sampled during data acquisition. Large-scale applications of the HS remain limited due to the cost of this type of technology and the technical difficulties in hyperspectral data processing. Low-cost portable hyperspectral cameras (PHCs) have been progressively developed; however, critical aspects associated with data acquisition and processing, such as the presence of spectral discontinuities, signal jumps, and a high level of background noise, were reported. The aim of this work was to analyze and improve the hyperspectral output of a PHC Senop HSC-2 device by developing a general use methodology. Several signal gaps were identified as falls and jumps across the spectral signatures near 513, 650, and 930 nm, while the dark current signal magnitude and variability associated with instrumental noise showed an increasing trend over time. A data correction pipeline was successfully developed and tested, leading to 99% and 74% reductions in radiance signal jumps identified at 650 and 830 nm, respectively, while the impact of noise on the acquired signal was assessed to be in the range of 10% to 15%. The developed methodology can be effectively applied to other low-cost hyperspectral cameras.

The paper focuses on the seasonal oil accumulation in traditional and super-high density (SHD) olive plantations and its modelling employing image-based linear models. For these purposes, at 7-10-day intervals, fruit samples (cultivar Arbequina, Fasola, Frantoio, Koroneiki, Leccino, Maiatica) were pictured and images segmented to extract the Red (R), Green (G), and Blue (B) mean pixel values which were re-arranged in 35 RGB-derived colorimetric indexes ( CIs ). After imaging, the samples were crushed and oil concentration was determined (NIR). The analysis of the correlation between oil and CIs revealed a differential hysteretic behavior depending on the covariates ( CI and cultivar). The hysteresis area ( Hyst ) was then quantified and used to rank the CIs under the hypothesis that CIs with the maximum or minimum Hyst had the highest correlation coefficient and were the most suitable predictors within a general linear model. The results show that the predictors selected according to Hyst-based criteria had high accuracy as determined using a Global Performance Indicator (GPI) accounting for various performance metrics ( R 2 , RSME, MAE). The use of a general linear model here presented is a new computational option integrating current methods mostly based on artificial neural networks. RGB-based image phenotyping can effectively predict key quality traits in olive fruit supporting the transition of the olive sector towards a digital agriculture domain.

Production of value-added compounds from waste materials is of utmost importance for the development of a sustainable society especially regarding their use as catalysts in industrially relevant synthetic reactions. Herein, we show the production of laccases from four white-rot fungi, which were grown on agricultural residues, specifically Trametes versicolor  11269, Pleurotus ostreatus  1020, Panus tigrinus 707 and Lentinula edodes SC-495. The produced laccases were tested on a laccase-mediator system (LMS) for the biocatalytic oxidation of the model substrate benzyl alcohol into benzaldehyde. The LMS was carried out in the presence both of tetrahydrofuran as co-solvent and of the mediator 2,2,6,6-tetramethyl-1-piperidinyloxyl (TEMPO) due to its high redox potential and its ability to perform the oxidation. Tolerance studies showed that the dialyzed solutions were able to tolerate 1% (99:1 v/v) of co-solvent, whereas a concentration of 10% v/v had a detrimental activity. Performances in the biocatalytic oxidation of laccase solutions from different purification steps were compared. Similar conversion was observed for laccase in dialysis (raw) and gel filtration (GF) product versus commercial T. versicolor laccase. The latter oxidized almost 99% of substrate while the other laccase solutions were able to reach a conversion from 91% for the laccase solution from P. tigrinus 707 after dialysis, to 50% for the laccase solution from P. ostreatus 1020 after gel filtration. This work highlights the potential of unpurified laccase solutions to be used as catalysts in synthetic reactions.

Many plants can modify their leaf profile rapidly in response to environmental stress. Image-based data are increasingly used to retrieve reliable information on plant water status in a non-contact manner that has the potential to be scaled to high-throughput and repeated through time. This paper examined the variation of leaf angle as measured by both 3D images and goniometer in progressively drought stressed grapevine. Grapevines, grown in pots, were subjected to a 21-day period of drought stress receiving 100% (CTRL), 60% ( ) and 30% ( ) of maximum soil available water capacity. Leaf angle was (i) measured manually (goniometer) and (ii) computed by a 3D reconstruction method (multi-view stereo and structure from motion). Stomatal conductance, leaf water potential, fluorescence ( / ), leaf area and 2D RGB data were simultaneously collected during drought imposition. Throughout the experiment, values of leaf water potential ranged from -0.4 (CTRL) to -1.1 MPa ( ) and it linearly influenced the leaf angle when measured manually ( = 0.86) and with 3D image ( = 0.73). Drought was negatively related to stomatal conductance and leaf area growth particularly in while photosynthetic parameters (i.e., / ) were not impaired by water restriction. A model for leaf area estimation based on the number of pixels of 2D RGB images developed at a different phenotyping robotized platform in a closely related experiment was successfully employed ( = 0.78). At the end of the experiment, top view 2D RGB images showed a ∼50% reduction of greener fraction (GGF) in CTRL and vines compared to initial values, while GGF in increased by approximately 20%.

Kiwifruit species have a relatively high rate of root oxygen consumption, making them very vulnerable to low root zone oxygen concentrations resulting from soil waterlogging. Recently, kiwifruit rootstocks have been increasingly used to improve biotic and abiotic stress tolerance and crop performance under adverse conditions. The aim of the present study was to evaluate morpho-physiological changes in kiwifruit rootstocks and grafting combinations under short-term waterlogging stress. A pot trial was conducted at the ALSIA PhenoLab, part of the Phen-Italy infrastructures, using non-destructive RGB and NIR image-based analysis and physiological measurements to identify waterlogging stress indicators and more tolerant genotypes. Three pot-grown kiwifruit rootstocks ('Bounty 71,' -B; 'D1,' var. -D; and 'Hayward,' var. -H) and grafting combinations, with a yellow-fleshed kiwifruit cultivar ('Zesy 002,' var. ) grafted on each rootstock (Z/B, Z/D, Z/H), were subjected to a control irrigation treatment (WW), restoring their daily water consumption, and to a 9-day waterlogging stress (WL), based on substrate saturation. Leaf gas exchange, photosynthetic activity, leaf temperature, RGB, and NIR data were collected during waterlogging stress. Stomatal conductance and transpiration reached very low values (less than 0.05 mol m s and 1 mmol m s , respectively) in both waterlogged D and H rootstocks and their grafting combinations. In turn, leaf temperature was significantly increased and photosynthesis was reduced (1-6 μmol m s ) from the first days of waterlogging stress compared to B rootstock and combination. The B rootstock showed prolonged leaf gas exchange and photosynthetic activity, indicating that it can cope with short-term and temporary waterlogging and improve the tolerance of grafted kiwi vines, which showed a decrease in stomatal conductance 5 days after the onset of stress. Morphometric and colorimetric parameters from the image-based analysis confirmed the greater susceptibility of D and H rootstocks and their grafting combinations to waterlogging stress compared to B. The results presented confirm the role of physiological measurements and enhance that of RGB and NIR images in detecting the occurrence of water stress and identifying more tolerant genotypes in kiwifruit.

Background In the last decade, the availability of gene sequences of many plant species, including tomato, has encouraged the development of strategies that do not rely on genetic transformation techniques (GMOs) for imparting desired traits in crops. One of these new emerging technology is TILLING (Targeting Induced Local Lesions In Genomes), a reverse genetics tool, which is proving to be very valuable in creating new traits in different crop species. Results To apply TILLING to tomato, a new mutant collection was generated in the genetic background of the processing tomato cultivar Red Setter by treating seeds with two different ethylemethane sulfonate doses (0.7% and 1%). An associated phenotype database, LycoTILL, was developed and a TILLING platform was also established. The interactive and evolving database is available online to the community for phenotypic alteration inquiries. To validate the Red Setter TILLING platform, induced point mutations were searched in 7 tomato genes with the mismatch-specific ENDO1 nuclease. In total 9.5 kb of tomato genome were screened and 66 nucleotide substitutions were identified. The overall mutation density was estimated and it resulted to be 1/322 kb and 1/574 kb for the 1% EMS and 0.7% EMS treatment respectively. Conclusions The mutation density estimated in our collection and its comparison with other TILLING populations demonstrate that the Red Setter genetic resource is suitable for use in high-throughput mutation discovery. The Red Setter TILLING platform is open to the research community and is publicly available via web for requesting mutation screening services.

In prostate cancer treatment with external beam radiation therapy (EBRT), prostate motion and internal changes in tissue distribution can lead to a decrease in plan quality. In most currently used planning methods, the uncertainties due to prostate motion are compensated by irradiating a larger treatment volume. However, this could cause underdosage of the treatment volume and overdosage of the organs at risk (OARs). To reduce this problem, in this proof of principle study we developed and evaluated a novel adaptive planning method. The strategy proposed corrects the dose delivered by each beam according to the actual position of the target in order to produce a final dose distribution dosimetrically as similar as possible to the prescribed one. Our adaptive planning method was tested on a phantom case and on a clinical case. For the first, a pilot study was performed on an in-silico pelvic phantom. A \"library\" of intensity modulated RT (IMRT) plans corresponding to possible positions of the prostate during a treatment fraction was generated at planning stage. Then a 3D random walk model was used to simulate possible displacements of the prostate during the treatment fraction. At treatment stage, at the end of each beam, based on the current position of the target, the beam from the library of plans, which could reproduce the best approximation of the prescribed dose distribution, was selected and delivered. In the clinical case, the same approach was used on two prostate cancer patients: for the first a tissue deformation was simulated in-silico and for the second a cone beam CT (CBCT) taken during the treatment was used to simulate an intra-fraction change. Then, dosimetric comparisons with the standard treatment plan and, for the second patient, also with an isocenter shift correction, were performed. For the phantom case, the plan generated using the adaptive planning method was able to meet all the dosimetric requirements and to correct for a misdosage of 13% of the dose prescription on the prostate. For the first clinical case, the standard planning method caused underdosage of the seminal vesicles, respectively by 5% and 4% of the prescribed dose, when the position changes for the target were correctly taken into account. The proposed adaptive planning method corrected any possible missed target coverage, reducing at the same time the dose on the OARs. For the second clinical case, both with the standard planning strategy and with the isocenter shift correction target coverage was significantly worsened (in particular uniformity) and some organs exceeded some toxicity objectives. While with our approach, the most uniform coverage for the target was produced and systematically the lowest toxicity values for the organs at risk were achieved. In our proof of principle study, the adaptive planning method performed better than the standard planning and the isocenter shift methods for prostate EBRT. It improved the coverage of the treatment volumes and lowered the dose to the OARs. This planning method is particularly promising for hypofractionated IMRT treatments in which a higher precision and control on dose deposition are needed. Further studies will be performed to test more extensively the proposed adaptive planning method and to evaluate it at a full clinical level.

Aiding optimal plant-environment interaction would favor plant resilience against environmental constrains including salt stress. We test the hypothesis that 6-Benzylaminopurine (BAP) primes grapevine's salt tolerance in vines (Vitis vinifera) received salt water (NaCl 100 mM) through the modulation of gene expression of BAP (AHK4, AHP1) and salt-stress (CAT, APX) inducible genes and morpho-physiological traits. A subgroup of vines had previously (48 h) been primed with BAP (80 mg/L) before salt stress. The gene expressions were 30% (CAT) and 56% (APX) lower in primed salt-stressed vines than that in un-primed. Salt treatment did not increase leaf Na + but it lowered stomatal conductance (g s ), photosynthesis (A), stem water potential (less negative) and photosystem-II efficiency (F v /F m ). Chlorophyll-a concentrations were 30% higher in BAP-primed compared to un-primed. Adverse effects of salt were significantly reduced, maintaining high A/g s , F v /F m and growth. After the relief of the stress, the BAP primed vines had a fast recovery.