A Genomic BSAseq Approach for the Characterization of QTLs Underlying Resistance to Fusarium oxysporum in Eggplant.

Eggplant (Solanum melongena L.), similar to many other crops, suffers from soil-borne diseases, including Fusarium oxysporum f. sp. melongenae (Fom), causing wilting and heavy yield loss. To date, the genetic factors underlying plant responses to Fom are not well known. We previously developed a Recombinant Inbred Lines (RILs) population using as a female parent the fully resistant line '305E40' and as a male parent the partially resistant line '67/3'. The fully resistant trait to Fom was introgressed from the allied species S. aethiopicum. In this work, the RIL population was assessed for the responses to Fom and by using a genomic mapping approach, two major QTLs on chromosomes CH02 and CH11 were identified, associated with the full and partial resistance trait to Fom, respectively. A targeted BSAseq procedure in which Illumina reads bulks of RILs grouped according to their resistance score was aligned to the appropriate reference genomes highlighted differentially enriched regions between resistant/susceptible progeny in the genomic regions underlying both QTLs. The characterization of such regions allowed us to identify the most reliable candidate genes for the two resistance traits. With the aim of revealing exclusive species-specific contigs and scaffolds inherited from the allied species and thus associated with the full resistance trait, a draft de-novo assembly of available Illumina sequences of the '305E40' parent was developed to better resolve the non-recombining genomic region on its CH02 carrying the introgressed Fom resistance locus from S. aethiopicum.

Less is more: natural variation disrupting a miR172 gene at the di locus underlies the recessive double-flower trait in peach (P. persica L. Batsch).

BACKGROUND: With the domestication of ornamental plants, artificial selective pressure favored the propagation of mutations affecting flower shape, and double-flower varieties are now readily available for many species. In peach two distinct loci control the double-flower phenotype: the dominant Di2 locus, regulated by the deletion of the binding site for miR172 in the euAP2 PETALOSA gene Prupe.6G242400, and the recessive di locus, of which the underlying factor is still unknown. RESULTS: Based on its genomic location a candidate gene approach was used to identify genetic variants in a diverse panel of ornamental peach accessions and uncovered three independent mutations in Prupe.2G237700, the gene encoding the transcript for microRNA miR172d: a ~5.0 Kb LTR transposable element and a ~1.2 Kb insertion both positioned upstream of the sequence encoding the pre-miR172d within the transcribed region of Prupe.2G237700, and a ~9.5 Kb deletion encompassing the whole gene sequence. qRT-PCR analysis confirmed that expression of pre-miR172d was abolished in di/di genotypes homozygous for the three variants. CONCLUSIONS: Collectively, PETALOSA and the mutations in micro-RNA miR172d identified in this work provide a comprehensive collection of the genetic determinants at the base of the double-flower trait in the peach germplasms.

A SmelAAT Acyltransferase Variant Causes a Major Difference in Eggplant (Solanum melongena L.) Peel Anthocyanin Composition.

Eggplant berries are rich in anthocyanins like delphinidin-3-rutinoside (D3R) and nasunin (NAS), which are accumulated at high amounts in the peel. NAS is derived by D3R through acylation and glycosylation steps. The presence of D3R or NAS is usually associated with black-purple or lilac fruit coloration of the most cultivated varieties, respectively. Building on QTL mapping position, a candidate gene approach was used to investigate the involvement of a BAHD anthocyanin acyltransferase (SmelAAT) in determining anthocyanin type. The cDNA sequence comparison revealed the presence of a single-base deletion in D3R-type line '305E40' (305E40_aat) with respect to the NAS-type reference line '67/3'. This is predicted to cause a frame shift mutation, leading to a loss of SmelAAT function and, thus, D3R retention. RT-qPCR analyses confirmed SmelAAT and 305E40_aat expression during berry maturation. In D3R-type lines, '305E40' and 'DR2', overexpressing the functional SmelAAT allele from '67/3', the transcript levels of the transgene correlated with the accumulation of NAS in fruit peel. Furthermore, it was also found a higher expression of the transcript for glucosyltransferase Smel5GT1, putatively involved with SmelAAT in the last steps of anthocyanin decoration. Finally, an indel marker matching with anthocyanin type in the '305E40' × '67/3' segregating population was developed and validated in a wide number of accessions, proving its usefulness for breeding purposes.

Scrapped but not neglected: Insights into the composition, molecular modulation and antioxidant capacity of phenols in peel of eggplant (Solanum melongena L.) fruits at different developmental stages.

Eggplant fruits are normally harvested and marketed when they reach the commercial maturity, that precedes the physiological ripening when dramatic changes in taste, composition and peel color take place. The biochemical changes in fruit peel across the developmental stages, characterized also by a sizeable decrement of anthocyanins, were studied in four eggplant genotypes differing for fruit pigmentation. HPLC-DAD, HPLC-ESI-MS and NMR analyses identified naringenin chalcone and naringenin 7-O-glucoside as the main phenolic compounds in extracts from the physiological ripe stage, along with compounds tentatively identified as glycosylated naringenin chalcone, naringenin and kaempferol. On average, the levels of anthocyanins, responsible for the peel pigmentation, dropped by 75% during development, while, surprisingly, the level of total phenols showed a slight decrease of 16%, with a final concentration of more than 1000 mg/100g dw. RT-qPCR expression profiling of nine genes coding for enzymes putatively acting at different steps of the involved pathways showed modulation mostly consistent with the observed changes in phenolic composition, with a remarkable decrease in the activity of flavonol reductase and an increase in flavonol synthase during berry development. Antioxidant activity monitored by peroxyl scavenging was similar at all developmental stages while Fremy's analysis evidenced a slight decrement at full physiological ripening. These results are valuable to address the improvement of eggplant commercial fruit quality and the valorization of unmarketable physiological ripe fruits, especially for the newly accumulation of the health-promoting compounds chalcones and flavanones.

Long-term follow-up of the Derivo® Embolization Device (DED®) for intracranial aneurysms: the Italian Multicentric Registry.

BACKGROUND: The flow-diverter devices (FDDs) safety and effectiveness have been demonstrated by large series and meta-analyses. Due to the high occlusion rates and the acceptable morbidity rates of FDDs, the indications for their use are continuously expanding. We presented our Italian multicentric experience using the second generation of DERIVO® Embolization Device (DED®; Acandis, Pforzheim, Germany) to cure cerebral aneurysms, evaluating both middle and long-term safety and efficacy of this device. METHODS: Between July 2016 and September 2017 we collected 109 consecutive aneurysms in 108 patients treated using DED® during 109 endovascular procedures in 34 Italian centers (100/109 aneurysms were unruptured, 9/109 were ruptured). The collected data included patient demographics, aneurysm location and characteristics, baseline angiography, adverse event and serious adverse event information, morbidity and mortality rates, and pre- and post-treatment modified Rankin Scale scores. Midterm and long-term clinical, angiographic and cross-sectional CT/MR follow-up were recorded and collected until December 2018. RESULTS: In 2/109 cases, DED® placement was classified as technical failures. The overall mortality and morbidity rates were respectively 6.5% and 5.5%. Overall DERIVO® related mortality and morbidity rates were respectively 0% and 4.6% (5 out of 108 patients). Midterm neuroimaging follow-up showed the complete or nearly complete occlusion of the aneurysm in 90% cases, which became 93% at long-term follow-up. Aneurysmal sac shrinking was observed in 65% of assessable aneurysms. CONCLUSIONS: Our multicentric experience using DED® for endovascular treatment of unruptured and ruptured aneurysms showed a high safety and efficacy profile, substantially equivalent or better compared to the other FDDs.

Identification of a new R3 MYB type repressor and functional characterization of the members of the MBW transcriptional complex involved in anthocyanin biosynthesis in eggplant (S. melongena L.).

Here we focus on the highly conserved MYB-bHLH-WD repeat (MBW) transcriptional complex model in eggplant, which is pivotal in the transcriptional regulation of the anthocyanin biosynthetic pathway. Through a genome-wide approach performed on the recently released Eggplant Genome (cv. 67/3) previously identified, and reconfirmed by us, members belonging to the MBW complex (SmelANT1, SmelAN2, SmelJAF13, SmelAN1) were functionally characterized. Furthermore, a regulatory R3 MYB type repressor (SmelMYBL1), never reported before, was identified and characterized as well. Through a qPCR approach, we revealed specific transcriptional patterns of candidate genes in different plant tissue/organs at two stages of fruit development. Two strategies were adopted for investigating the interactions of bHLH partners (SmelAN1, SmelJAF13) with MYB counterparts (SmelANT1, SmelAN2 and SmelMYBL1): Yeast Two Hybrid (Y2H) and Bimolecular Fluorescent Complementation (BiFC) in A. thaliana mesophylls protoplast. Agro-infiltration experiments highlighted that N. benthamiana leaves transiently expressing SmelANT1 and SmelAN2 showed an anthocyanin-pigmented phenotype, while their co-expression with SmelMYBL1 prevented anthocyanin accumulation. Our results suggest that SmelMYBL1 may inhibits the MBW complex via the competition with MYB activators for bHLH binding site, although this hypothesis requires further elucidation.

Trends in utilization of transarterial treatments for hepatocellular carcinoma: results of a survey by the Italian Society of Interventional Radiology.

PURPOSE: This study was designed to provide an overview of the practice of locoregional treatments for HCC by the Italian centers of Interventional Radiology (IR) with particular reference to transarterial modalities. METHODS: A questionnaire of 11 questions on locoregional treatment of HCC was e-mailed to 134 Italian IR centers. RESULTS: The response rate was 64.9% (87/135 centers). Of 8,959 procedures in 2011, 67% were transarterial treatments, 31% percutaneous ablations, and 2% Y90-radioembolizations. Regarding (chemo)embolization, approximately 59% of procedures were performed in the intermediate stage, 28% in the early stage, and 12.8% in the advanced stage. TACE techniques varied greatly; approximately 52% of procedures were performed with drug-eluting particles and 32% with lipiodol, drug, and reabsorbable particles. In selected cases, 53 of 78 (68%) centers combine chemoembolization and ablation, whereas 28 centers (35.9%) combine Sorafenib and chemoembolization. In 2011, 13 of 78 (16.7%) responding centers performed Y90-radioembolization, with approximately 52% of procedures performed in the advanced stage and 46% in the intermediate stage. Approximately 62% of Y90-radioembolizations were performed using resin spheres and 38% using glass spheres. CONCLUSIONS: With almost 9,000 procedures performed each year, locoregional treatments of HCC, most of all transarterial (chemo)embolizations, represent a major part of daily clinical practice in many Italian IR centers. The high variability in responses regarding transarterial treatments for HCC patients highlights the need for solid scientific evidence allowing better definition of clinical indications and standardization of technical approaches.

Preoperative assessment of stenosis of the epiaortic vessels: can colour-Doppler ultrasound really supplant angiography?

PURPOSE: To evaluate the diagnostic role of digital angiography compared with colour-Doppler US in the study of epiaortic vessel stenoses and the degree of morbidity associated with angiography. MATERIALS AND METHODS: Out of 2,000 angiographic examinations of the epiaortic vessels performed for the concurrent presence of clinical findings of cerebrovascular insufficiency and colour-Doppler US findings of carotid-vertebral stenosis, a randomized sample of 200 patients was evaluated. The retrospective review of medical records provided the surgical data of the stenosis which were subsequently compared with the angiography and colour-Doppler US findings, and with the clinical and laboratory data to identify the possible complications of angiography. RESULTS: Diagnostic agreement was 75%. Colour-Doppler US underestimated stenosis in 16.5% and overestimated stenosis in 8% of cases. In 18% of cases there was a diagnostic gain of angiography (vascular lesions associated with main lesion), which also provided clinically important incidental findings (intracranial aneurysms, meningiomas) in 6.5% of patients. No relevant complications resulting from the angiographic procedure were recorded. CONCLUSIONS: Angiography allowed a more accurate evaluation of the degree of stenosis compared with colour-Doppler ultrasound. Moreover, it allowed diagnosis of important associated conditions that may affect therapeutic planning. In our series, we recorded no important complications related to angiography. Therefore, angiography remains the standard of reference in the preoperative evaluation of patients with clinically diagnosed cerebral ischaemia. Though fairly reliable as a first-line investigation, Colour-Doppler US is not sufficiently exhaustive as a pre-operative evaluation tool.

Actual role of unenhanced magnetic resonance angiography (mra tof 3d) in the study of stenosis and occlusion of extracranial carotid artery.

PURPOSE: To evaluate the diagnostic accuracy, sensitivity and specificity of MRA (3D TOF) in comparison to contrast-enhanced MRA using DSA as the gold standard in the study of steno-occlusive disease of the internal carotid artery, in order to determine if this technique still has a diagnostic value. MATERIALS AND METHODS: Three radiologists blindly and independently evaluated either the unenhanced MRA, CE MRA, or DSA images of 23 consecutive patients, assessing the stenosis based on a NASCET five-class classification. RESULTS: The sensitivity, specificity, and diagnostic accuracy of CE MRA and unenhanced MRA were 100% and 100%, 100% and 95.5% and 100% and 93.2%, respectively. These differences were not statistically significant. CONCLUSIONS: Unenhanced MRA should be still considered a valid alternative for studying the extracranial section of the internal carotid arteries especially in patients who refuse or whose laboratory results exclude the use of contrast medium or in patients in whom continuous monitoring of a clinical condition is required but a repeated and/or frequent use of contrast medium is not possible.

Endovascular treatment of intracranial arterio-venous malformations with Onyx embolization: preliminary experience.

PURPOSE: Intracranial arterio-venous malformations (AVM) often represent a complex clinical problem as regards indications to treatment, the choice of treatment and the technical difficulties related to treatment. In the last twenty years, endovascular treatment of intracranial arterio-venous malformations was primarily based on an acrylic glue (Hystoacril) whose endovascular use is not free from risks. A new product for endovascular embolization, named ONYX, has been recently been made commercially available. It is a bio-compatible liquid polymer that precipitates and solidifies in contact with blood, thus forming a soft and spongy embolus. The aim of this study was to evaluate our preliminary results in the endovascular embolization of intracranial AVM with Onyx. MATERIALS AND METHODS: Ten patients were treated (7 men, 3 women; mean age: 29 years, range: 12-48 years) for a total of 37 embolizations, 22 with Onyx and 15 with acrylic resin. RESULTS: Nidus occlusion was complete in 2 cases, >90% in 2 cases, >50 and <90% in 3 cases and <50% in the remaining 3 cases. The following complications were recorded: two transient and one mild permanent neurological deficits, two clinically silent cases of moderate subarachnoid haemorrhage, four catheters glued to the injection site, three cases of treatment discontinuation due to continuous and massive reflux of Onyx into the afferent artery peduncle. CONCLUSIONS: This early experience showed that while Onyx has good embolization potential it also presents some disadvantages, which need to be overcome before this product can be considered easy and safe to use on a large scale.