TRANSCRIPTOME CHARACTERIZATION AND TRANSCRIPTIONAL REPROGRAMMING OF ZUCCHINI PLANT IN RESPONSE TO APHIS GOSSYPII FEEDING

Zucchini (Cucurbita pepo L.) belongs to the Cucurbitaceae family and ranks among the highest-valued vegetables worldwide. It is widely cultivated in temperate region where one of the main problem related to its cultivation is the damage imposed by the cotton/melon aphid Aphis gossypii (Homoptera: Aphididae). Aphis gossypii is a polyphagous aphid which can both directly and indirectly affects host plant by inducing leaf curling and necrosis and vectoring several plant viruses. Plants can defend themselves against aphids using different strategies. A direct response activates a deep transcriptional reprogramming which leads to de novo synthesis of proteins and molecules implicated in defence, including production of herbivore induced volatile organic compounds (HI-VOCs), which can indirectly attract herbivore natural enemies. In the present study, the mRNA from un-infested (control) and infested leaves by A. gossypii of C. pepo cultivar “San Pasquale” was sequenced to obtain a de novo transcriptome assembly to be used as reference for gene expression profiling. Leaf material was collected from control and infested (10 adult A. gossypii) plants at three different time points (24, 48, 96 hours post infection; hpi) and RNA was extracted. Illumina sequencing generated ~34 million of paired-end reads of 101 nucleotides in length per sample. Short reads were pre-processed and, then, de novo assembled using Velvet/Oases and CAP3 tools into a non-redundant set of 71,648 transcripts. Approximately 94% of the assembled transcripts contains coding sequences that could be translated into proteins, and ~70% of transcripts was successfully annotated using BLAST similarity-based searches and Blast2GO. Furthermore, BLASTn comparisons with the publically available C. pepo transcriptome resulted in 1,313 transcripts exclusively assembled in the aphid-challenged transcriptome. Following transcriptome assembly, a dataset of 42,517 sequences, in which each gene locus was represented only once, was filtered out and used as reference for read mapping and differentially expressed gene (DEG) call. A total of 766 transcripts was differentially expressed (FDR< 0.05; -2˂logFC˃2). At 24 hpi, 158 transcripts were influenced by aphid infestation. The number of affected transcripts increased to 565 at 48 hpi and declined to 179 transcripts at 96 hpi. The analysis of DEGs highlighted the modulation of genes involved in hormone-related defence pathways. Among these, SA-related genes were found mainly up-regulated assuming an important role in “San Pasquale”-A. gossypii interaction. Furthermore, a significant negative effect on aphid fixing behaviour was observed on zucchini plants pre-treated with synthetic methyl salicylate (MeSA). Zucchini plant response was also characterised by the overexpression of genes involved in primary metabolic processes as well as cell wall modification. Interestingly, several aphid-derived transcripts were discovered among zucchini DEGs. Even if no conclusive evidence can be drawn, we hypothesized that these mRNAs might play a role in modulation of plant direct and/or indirect response. Finally, analysis of VOCs emitted by zucchini plants infested with 10 adult aphids, for 48 h, showed a significant reduction in (E)-caryophyllene emission, whereas emission levels of other volatiles were not affected. Conversely, a significant increase in (E)-caryophyllene emission was observed when plants were infested with 300 adult aphids, for 96 h. Moreover, olfactometer bioassays revealed that synthetic (E)-caryophyllene was attractive to female Aphidius colemani parasitic waps, widely used as biological control agents for A. gossypii. Taken together, these results suggest (E)-caryophyllene may play an important role in zucchini plant indirect defence responses. Our study allows to elucidate, for the first time, the molecular mechanisms activated by zucchini plants in response to A. gossypii infestation during a compatible interaction

De Novo Transcriptome Assembly of Cucurbita Pepo L. Leaf Tissue Infested by Aphis Gossypii

Zucchini (Cucurbita pepo L.), extensively cultivated in temperate areas, belongs to the Cucurbitaceae family and it is a species with great economic value. One major threat related to zucchini cultivation is the damage imposed by the cotton/melon aphid Aphis gossypii Glover (Homoptera: Aphididae). We performed RNA-sequencing on cultivar "San Pasquale" leaves, uninfested and infested by A. gossypii, that were collected at three time points (24, 48, and 96 h post infestation). Then, we combined all high-quality reads for de novo assembly of the transcriptome. This resource was primarily established to be used as a reference for gene expression studies in order to investigate the transcriptome reprogramming of zucchini plants following aphid infestation. In addition, raw reads will be valuable for new experiments based on the latest bioinformatic tools and analytical approaches. The assembled transcripts will serve as an important reference for sequence-based studies and for primer design. Both datasets can be used to support/improve the prediction of protein-coding genes in the zucchini genome, which has been recently released into the public domain

Isolated slaughterhouse liver as model for normothermic perfusion after warm and cold ischemia: single case report

AbstractLiver transplantation is an ultimate procedure in patients suffering end-stage liver diseases. In these last years the donation after cardiac death (DCD) has increased the pool of potential liver donors. Different studies and procedures are involved in the prevention of the main ischemic problems during the reconditioning and resuscitation of the marginal livers. Normothermic extracorporeal liver perfusion (NELP) avoids prolonged cold storage damage that is the main cause of steatosis and biliary tract ischemia in transplanted patiens. Different porcine models have been studied and developed to understand the ischemia mechanism and to select the better technique for NELP.We conducted our study using a DCD pig liver model collected from slaughterhouse. Using extracorporeal membrane oxygenation, 2000 ml of total fluid containing autologous blood, lidocaine, heparin, antibiotics, glucose 10 % solution and flunixin, the NELP was achieved. The liver was perfused over 7 hours after 48 hours of cold storage (4C°), using Eurocollins solution. During the liver withdrawal in the slaughterhouse 20 minutes were waited to simulate the warm ischemia (WI) time. Histological samples, swab for bacterial grow, blood sample, temperature and pulse oximetry saturation were collected to assess the liver viability and function. These analyses revealed stable metabolism throughout perfusion identifying a cycles 2 hours length, coinciding with recovery of oxygen uptake rates to fresh liver, as described in literature.In summary the preliminary established model of isolated hemoperfused slatherhouse liver reveals the important role of the relation between cold storage and normothermic perfusion. Moreover this preliminary study justifies further investigation of the optimization of the treatment protocols and perfusion media

How future surgery will benefit from SARS-COV-2-related measures: a SPIGC survey conveying the perspective of Italian surgeons

COVID-19 negatively affected surgical activity, but the potential benefits resulting from adopted measures remain unclear. The aim of this study was to evaluate the change in surgical activity and potential benefit from COVID-19 measures in perspective of Italian surgeons on behalf of SPIGC. A nationwide online survey on surgical practice before, during, and after COVID-19 pandemic was conducted in March-April 2022 (NCT:05323851). Effects of COVID-19 hospital-related measures on surgical patients' management and personal professional development across surgical specialties were explored. Data on demographics, pre-operative/peri-operative/post-operative management, and professional development were collected. Outcomes were matched with the corresponding volume. Four hundred and seventy-three respondents were included in final analysis across 14 surgical specialties. Since SARS-CoV-2 pandemic, application of telematic consultations (4.1% vs. 21.6%; p &lt; 0.0001) and diagnostic evaluations (16.4% vs. 42.2%; p &lt; 0.0001) increased. Elective surgical activities significantly reduced and surgeons opted more frequently for conservative management with a possible indication for elective (26.3% vs. 35.7%; p &lt; 0.0001) or urgent (20.4% vs. 38.5%; p &lt; 0.0001) surgery. All new COVID-related measures are perceived to be maintained in the future. Surgeons' personal education online increased from 12.6% (pre-COVID) to 86.6% (post-COVID; p &lt; 0.0001). Online educational activities are considered a beneficial effect from COVID pandemic (56.4%). COVID-19 had a great impact on surgical specialties, with significant reduction of operation volume. However, some forced changes turned out to be benefits. Isolation measures pushed the use of telemedicine and telemetric devices for outpatient practice and favored communication for educational purposes and surgeon-patient/family communication. From the Italian surgeons' perspective, COVID-related measures will continue to influence future surgical clinical practice

Plant defense strategies against attack by multiple herbivores

Plants may effectively tailor defenses by recognizing their attackers and reprogramming their physiology. Although most plants are under attack by a large diversity of herbivores, surprisingly little is known about the physiological capabilities of plants to deal with attack by multiple herbivores. Studies on dual herbivore attack identified that defense against one attacker may cause energetic and physiological constraints to deal with a second attacker. How these constraints shape plant plasticity in defense to their full community of attackers is a major knowledge gap in plant science. Here, we provide a framework for plant defense to multiherbivore attack by defining the repertoire of plastic defense strategies that may allow plants to optimize their defenses against a multitude of stressors

Optic Nerve Ultrasound Evaluation in Children: A Review

Managing patients with neurocritical illness requires monitoring and treating elevated intracranial pressure (ICP), especially in cases in children. In terms of precise and real-time measurements, invasive ICP measurements are presently the gold standard for the initial diagnosis and follow-up ICP assessments. As a rapid and non-invasive way to detect elevated ICP, point-of-care ultrasonography (POCUS) of optic nerve sheath diameter (ONSD) has been proposed. The utility of bedside POCUS of ONSD to detect elevated ICP with excellent diagnostic test accuracy in adults has already been demonstrated. Nonetheless, data on the relationship between POCUS of ONSD and ICP in children are scarce. Therefore, the purpose of this review is to point out the most recent findings from the pediatric published literature and briefly discuss what was assessed with ONSD ultrasound examination, and also to describe and discuss the diagnostic procedures available for optic nerve ultrasound appraisal. A search of the medical databases PubMed and Scopus was carried out. The terms such as “ocular ultrasonography”, “ICP assessment”, “children”, “point-of-care ultrasound”, and “POCUS” were searched. In conclusion, the use of the standardized A-scan technique coupled with the B-scan technique should be suggested to provide data that are as accurate, precise, repeatable, and objective as possible

Bench to bedside review of myositis autoantibodies

Abstract Idiopathic inflammatory myopathies represent a heterogeneous group of autoimmune diseases with systemic involvement. Even though numerous specific autoantibodies have been recognized, they have not been included, with the only exception of anti-Jo-1, into the 2017 Classification Criteria, thus perpetuating a clinical-serologic gap. The lack of homogeneous grouping based on the antibody profile deeply impacts the diagnostic approach, therapeutic choices and prognostic stratification of these patients. This review is intended to highlight the comprehensive scenario regarding myositis-related autoantibodies, from the molecular characterization and biological significance to target antigens, from the detection tools, with a special focus on immunofluorescence patterns on HEp-2 cells, to their relative prevalence and ethnic diversity, from the clinical presentation to prognosis. If, on the one hand, a notable body of literature is present, on the other data are fragmented, retrospectively based and collected from small case series, so that they do not sufficiently support the decision-making process (i.e. therapeutic approach) into the clinics

A cross-sectional survey on dietary supplements consumption among italian teen-agers

Introduction: In the last decades, dietary supplements consumption has increased in the Western world for all age groups. The long-term potentially dangerous effects related to an indiscriminate consumption of dietary supplements are still unknown and are becoming a matter of public health concern. Therefore, the aims of the present study were: to explore the contribution of dietary supplements to micronutrient daily intake, and to evaluate awareness and knowledge about dietary supplements. Methods: Participants (age ranging from 14 to 18 years) were recruited among students attending 8 high schools in the province of Frosinone (Italy). An anonymous questionnaire, composed of 12 multiple-choice items, was administered to all the participants. T-test and the analysis of variance (ANOVA) were performed to describe differences between means of the groups, while the chi-square test was used to compare observed and expected frequencies. The logistic regression model, aimed at identifying the characteristics of potential consumers of dietary supplements, Results: A total of 686 teenagers (288 males and 398 females, average age: 17,89∓0,91 years) participated in the study. The 83,6% of participants affirmed to be aware of dietary supplements. 239 participants consumed dietary supplements: 118 males and 121 females. 49,1% of females consumed dietary supplements less than twice a week, whereas 43,6% of males consumed dietary supplements from 2 to 5 times per week. Statistically significant differences emerged between the genders with respect to the source of information regarding dietary supplements, the perceived indications for their use, and the choice of the store/place to purchase them. Discussion: Findings in the present study provide insight into the consumption of dietary supplements among young Italians, highlighting the need to foster further awareness among adolescents about the correct use of dietary supplements, especially in terms of indications and contraindications. © 2014 del Balzo et al

A cross-sectional survey on dietary supplements consumption among Italian teen-agers.

INTRODUCTION: In the last decades, dietary supplements consumption has increased in the Western world for all age groups. The long-term potentially dangerous effects related to an indiscriminate consumption of dietary supplements are still unknown and are becoming a matter of public health concern. Therefore, the aims of the present study were: to explore the contribution of dietary supplements to micronutrient daily intake, and to evaluate awareness and knowledge about dietary supplements. METHODS: Participants (age ranging from 14 to 18 years) were recruited among students attending 8 high schools in the province of Frosinone (Italy). An anonymous questionnaire, composed of 12 multiple-choice items, was administered to all the participants. T-test and the analysis of variance (ANOVA) were performed to describe differences between means of the groups, while the chi-square test was used to compare observed and expected frequencies. The logistic regression model, aimed at identifying the characteristics of potential consumers of dietary supplements. RESULTS: A total of 686 teenagers (288 males and 398 females, average age: 17,89±0,91 years) participated in the study. The 83,6% of participants affirmed to be aware of dietary supplements. 239 participants consumed dietary supplements: 118 males and 121 females. 49,1% of females consumed dietary supplements less than twice a week, whereas 43,6% of males consumed dietary supplements from 2 to 5 times per week. Statistically significant differences emerged between the genders with respect to the source of information regarding dietary supplements, the perceived indications for their use, and the choice of the store/place to purchase them. DISCUSSION: Findings in the present study provide insight into the consumption of dietary supplements among young Italians, highlighting the need to foster further awareness among adolescents about the correct use of dietary supplements, especially in terms of indications and contraindications