Study on ultra-structural effects caused by Onion yellow dwarf virus infection in ‘Rossa di Tropea’ onion bulb by means of magnetic resonance imaging

\u2018Rossa di Tropea\u2019 onion is a particular pink/red coloured onion cultivated in Calabria region (Southern Italy), representing one of the Italian most important vegetable crops granted with Protected Designation of Origin (PDO) and Protected Geographical Indication (PGI) trademarks. This local cultivar is characterised by a high nutraceutical compounds content showing anti-inflammatory, anti-cholesterol, anticancer and antioxidant properties. As all vegetable crops and Allium spp., \u2018Rossa di Tropea\u2019 onion is affected by several viruses. Among these, the species Onion yellow dwarf virus (OYDV, genus Potyvirus, family Potyviridae), represents the most limiting biotic stress, inducing severe symptoms. OYDV effect on tissues architecture in whole bulbs was investigated using magnetic resonance microimaging (MRI) technique, which allows the interior of samples to be imaged non-invasively and non-destructively and yields quantitative information on physico-chemical parameters describing water mobility (T1 and T2 relaxation times). The use of such tool allowed to determine how OYDV alters plant physiology by inducing water accumulation in bulb tissues as well as causing ultra-structural modifications of cell wall, highlighted by MRI. All these effects resulted in an increase of free water in plant tissues, and consequently relevant water losses during post-harvest storage, seriously affecting bulb quality, marketability and shelf life

WRKY Gene Family Drives Dormancy Release in Onion Bulbs

Onion (Allium cepa L.) is an important bulb crop grown worldwide. Dormancy in bulbous plants is an important physiological state mainly regulated by a complex gene network that determines a stop of vegetative growth during unfavorable seasons. Limited knowledge on the molecular mechanisms that regulate dormancy in onion were available until now. Here, a comparison between uninfected and onion yellow dwarf virus (OYDV)-infected onion bulbs highlighted an altered dormancy in the virus-infected plants, causing several symptoms, such as leaf striping, growth reduction, early bulb sprouting and rooting, as well as a lower abscisic acid (ABA) level at the start of dormancy. Furthermore, by comparing three dormancy stages, almost five thousand four hundred (5390) differentially expressed genes (DEGs) were found in uninfected bulbs, while the number of DEGs was significantly reduced (1322) in OYDV-infected bulbs. Genes involved in cell wall modification, proteolysis, and hormone signaling, such as ABA, gibberellins (GAs), indole3-acetic acid (IAA), and brassinosteroids (BRs), that have already been reported as key dormancyrelated pathways, were the most enriched ones in the healthy plants. Interestingly, several transcription factors (TFs) were up-regulated in the uninfected bulbs, among them three genes belonging to the WRKY family, for the first time characterized in onion, were identified during dormancy release. The involvement of specific WRKY genes in breaking dormancy in onion was confirmed by GO enrichment and network analysis, highlighting a correlation between AcWRKY32 and genes driving plant development, cell wall modification, and division via gibberellin and auxin homeostasis, two key processes in dormancy release. Overall, we present, for the first time, a detailed molecular analysis of the dormancy process, a description of the WRKY-TF family in onion, providing a better understanding of the role played by AcWRKY32 in the bulb dormancy release. The TF coexpressed genes may represent targets for controlling the early sprouting in onion, laying the foundations for novel breeding programs to improve shelf life and reduce postharvest

Validation of a microarrays protocol for detection and genotyping isolates of Plum pox virus

A genomic strategy for PPV identification has been recently developed (Pasquini et al., 2008). The method is based on using a 70-mer oligonucleotide DNA microarray chip capable of simultaneously detecting and genotyping PPV strains. Universal and specific probes have been identified and used with a sensitive protocol of hybridization using an indirect fluorescent labelling of cDNA product with cyanine able to enhance the sensitivity of the virus detection avoiding the use of the PCR amplification step. In order to evaluate the protocol fitness for diagnostic use, about 30 samples belonging to a PPV isolates collection, including M, D, EA and C strains, have been used for its validation, that was determined, estimating the performance criteria that include the following parameters: diagnostic sensitivity (D-SN), diagnostic specificity (D-SP) and diagnostic accuracy (D-AC). Keywords: oligonucleotides chip, PPV, sensitivity, specificity, accuracy, performance criteri

Eleven-year experience with the avidin-biotin pretargeting system in glioblastoma: Toxicity, efficacy and survival

Background: The 3-step avidin-biotin pretargeting approach is applied in patients with recurrent glioblastoma (GBM), using biotinylated anti-tenascin monoclonal antibody as the first step of pretargeting followed by avidin and 90Ybiotin. Methods: The present study reviews objective response and overall survival rates in 502 glioblastoma patients treated with 3-step radioimmunotherapy in our institute from December 1994 to December 2005. Patients underwent standard treatment before receiving Pretargeted Antibody-Guided Radionuclide Therapy with 90Y-biotin (PAGRIT ®). Results: Of the 502 patients, 272 (54%) were evaluable for response and 375 (75%) for overall survival. 174 patients (64%) continued to progress after PAGRIT ®, 77 (28%) obtained disease stabilization, and 21 (8%) showed objective tumor regression. Survival of the 375 evaluable patients was 98.4% at 6 months, 79.2% at 12 months, 51.7% at 18 months, and 30.7% at 24 months after the first cycle of PAGRIT ®. All 375 received 3-step PAGRIT ® at recurrence of GBM. The median survival time from diagnosis was 19 months. Conclusion: The results from this retrospective analysis suggest that 90Y-biotin PAGRIT ® interferes with the progression of glioblastoma, prolonging survival in a larger number of patients. Our analysis forms the basis for further prospective trials, where radioimmunotherapy, which is known to be more effective in minimal residual disease, could be offered immediately after surgery. © Grana et al.; Licensee Bentham Open

Applicability of brain oxygenation measurement in the assessment of post-concussion outcomes in combat sport

Purpose: Investigate the association in semi and professional boxers (mTBI), who experienced previous multiple sports concussions, between brain oxygenation, balance, maximal aerobic capacity and power, in comparison to non concussed (NC) subjects. Methods: We tested 6 mTBI age 20-43 years (1 female and 5 males) and 7 NC age 24-38 (3 females, 4 males) subjects. Cerebral changes in DO2Hb, DHHb and total hemoglobin were measured using a protocol of three steps (Rest before max VO2 test, Hypercapnia, and recovery after max VO2 test, 5 minutes each) with a Nimo Nirs system (Nirox, Brescia) and 02 measurement with a Cosmed Quark CPET (Cosmed, Roma). CVR (cerebral vascular reactivity) in mTBI and NC subjects and respiratory parameters were calculated. Aerobic fitness outcome was quantified by mean of VO2max with a Bruce test on a bike. A Romberg Test was performed on a force platform (Kistler 9281, Switzerland) and software Sway (BTS Eng., Milan). Analysis of transversal (ML) and Longitudinal (AP) range of oscillation was performed. A Fitt\u2019s test, was performed on a pc and scoring was obtained. Statistical analysis was performed with SPSS-v.23 (IBM Inc. Chicago) using the Mann-Whitney U test. Results: Any difference mTBI/NC was found in Fitt\u2019s test. Any differences were found between males and females in any of the parameters measured, except anthropometry with a weight of 0.29 for sex (GLM test, SPSS). VO2max for mTBI was 49.58 \ub1 5.19 and for NC was 47.47 \ub1 4.91 ml/kg/min-1 without any significant difference. mTBI (17.65 \ub1 4.79 mm) show a significant differences with NC (25.35 \ub1 4.11 mm) in medio-lateral sway with eyes open (sig. 0.008). At GLM test, major predictor was the medio-lateral sway with eyes open with a weight of 0.71. Also a significant was found difference of FetO2 average in hypercapnia (sig. 0.008). Conclusions: Concussion seems to be evidenced by medio-lateral sway values (eyes open) and FetO2 in hypercapnia. Reference 1. Amen DG et al. (2011) Reversing brain damage in former NFL players: implications for traumatic brain injury and substance abuse rehabilitation. Journal of Psychoactive Drugs 43: 1-

Loop mediated isothermal amplification: Principles and applications in plant virology

In the last decades, the evolution of molecular diagnosis methods has generated different advanced tools, like loop-mediated isothermal amplification (LAMP). Currently, it is a well-established technique, applied in different fields, such as the medicine, agriculture, and food industries, owing to its simplicity, specificity, rapidity, and low-cost efforts. LAMP is a nucleic acid amplification under isothermal conditions, which is highly compatible with point-of-care (POC) analysis and has the potential to improve the diagnosis in plant protection. The great advantages of LAMP have led to several upgrades in order to implement the technique. In this review, the authors provide an overview reporting in detail the different LAMP steps, focusing on designing and main characteristics of the primer set, different methods of result visualization, evolution and different application fields, reporting in detail LAMP application in plant virology, and the main advantages of the use of this technique

Complete sequence, genotyping and comparative analysis of Pepino mosaic virus discovered isolates from Italy

Pepino mosaic virus (PepMV) causes economic damage to the tomato industry worldwide. In Italy, the virus is endemic in Sicily, an important tomato-production region, and a new outbreak has been recorded in Sardinia after the first PepMV appearance in 2001. Italian PepMV isolates from these two geographically distant regions were molecularly characterized and the complete nucleotide sequence of four isolates was determined by walking primer strategy. The complete nucleotide and deduced amino acid sequences were analyzed and compared with published PepMV sequences. Italian isolates belong to the PepMV-Ch2 strain and had 97.2% to 98.5% identity with those of the reference PepMV-Ch2 deposited in GenBank. The two Sicilian isolates were slightly more variable in the RdRp and TGB1 genes. This study provides new insights into the occurrence and variability of PepMV isolates in Italy