Screening of local Italian maize varieties for resistance to Fusarium verticillioides

In order to find maize genetic sources of resistance to Fusarium attack and to fumonisin accumulation, 27 Italian local varieties collected at the CREA (Consiglio per la ricerca in agricoltura e l’analisi dell’economia agraria, Unità di ricerca per la maiscoltura) maize gene bank, were evaluated in artificial inoculation field experiments during 2011 and 2012 seasons. Primary ears were inoculated with a mixture of two Fusarium verticillioides toxigenic strains through kernel inocula- tion; non-inoculated and sterile water-inoculated ears were included as controls. Disease severity was estimated by counting the kernels with visible symptoms of infection at the inoculation point. The percentage of internally contaminated kernels was detected and fumonisin accumulation was recorded. Results showed that, under artifi- cial inoculation, genotypes responded differentially to the Fusarium attack. Five out of 27 genotypes (VA56, VA70, VA74, VA114, and VA121) showed a low fumonisin content both in 2011 and 2012, seasons characterized by very different climatic conditions

The b-32 ribosome inactivating protein from maize influences fumonisin accumulation in in-vitro bioassays

Fungi of the genus Fusarium are common plant pathogens mainly associated with cereal crops. In particular, Fusarium verticillioides Sacc. is the most common toxigenic fungus in maize worldwide, causing root, stalk, and ear rot. Fusarium spp. can produce a wide range of secondary metabolites, some of which can unfavourably affect human and animal health. Owing to the potential risks of fumonisins F. verticillioides secondary metabolites, new regulations for the allowable mycotoxin limit in food and feed have been put in place by most agencies worldwide. Plants act on the attack of pathogenic fungi through a complex network of active responses such as the production of proteins toxic or inhibitory to pathogens such as RIP (Ribosome-Inactivating-Protein). The RIP present in maize endosperm (termed b-32) has been widely investigated. Similarly to other RIPs, is accumulated in the seed as an inactive pro-RIP precursor, which is converted into an active form by proteolytic processing. To understand the relationships between structure and substrate specificity of the maize b-32 RIP protein, a series of recombinant b-32 sequences, by selective deleting of different domains (RIP b-32, RIP ∆N, RIP ∆C and RIP-∆C (Ala), were prepared. Recombinant sequences were expressed in Escherichia coli to obtain high levels of recombinant proteins, which were subsequently tested for their potential ability to reduce both the colonization of F. verticillioides and fumonisin accumulation

Characterization of the maize b-32 ribosome inactivating protein and its interaction with fungal pathogen development

Plants respond to attack by pathogenic fungi with a complex network of responses, including the production and accumulation of proteins, such as the Ribosome Inactivating Proteins (RIPs) that are toxic or inhibitory to pathogens. In maize endosperm, a cytosolic albumin termed b-32 (RIP1) is synthesized in temporal and quantita¬tive coordination with the deposition of storage proteins. Research has shown that b-32 is able to i) enzymati¬cally inactive ribosomes modifying rRNA inhibiting protein synthesis in vitro, ii) inhibit the growth of Rhizoctonia solani mycelia in an in vitro and in planta assays, iii) reduce Fusarium culmorum head blight in wheat transgenic plants expressing b-32, and iv) diminish Fusarium verticillioides attack symptoms in leaf tissues assays of maize transgenic expressing ectopically b-32 protein. Similarly to other RIPs, maize b-32 is accumulated in the seed as an inactive precursor, which is converted into an active form by proteolytic processing which removes peptide segments from the N (residues 1-16 of pro-RIP) and C (residues 295-301) termini and also from the center (linker domain) of the polypeptide. In this review we will summarize evidence and advances related to the ability of the b-32 protein in contrasting pathogen attacks by considering and describing i) in vivo b-32 antifungal activity and ii) in vitro fungal development inhibition. These data provide information for assessing b-32 in developing plants with a higher capacity to contrast damages induced by pathogens

Evaluation of ear rot (Fusarium verticillioides) resistance and fumonisin accumulation in Italian maize inbred lines

Mycotoxin contamination of maize (Zea mays L.) grain is a global threat to the safety of both human food and animal feed. Hence, the development of maize genotypes with reduced mycotoxin accumulation in grain is of major importance. In order to find maize germplasm sources of resistance to Fusarium ear rot, 34 Italian and six public inbred lines were evaluated by means of artificial inoculation in field experiments during 2009 and 2010. Relationships between ear rot and fumonisin concentration in the ears were investigated. Primary ears were challenged with a mixture of two Fusarium verticillioides isolates from Northern Italy, through kernel inoculation, and ear rot severity was assessed.The average number of visibly infected kernels per ear, after inoculation, ranged from 2 to 68 in 2009 and from 0 to 120 in 2010. Fumonisin concentrations in the inoculated ears were greater than in the experimental controls for both years. Variability was found between the inbred lines: fumonisin accumulation ranged from 0.56 to 240.83 mg kg-1 in 2009 and from 1.09 to 190.60 mg kg-1 in 2010. In both years, six inbred lines showed high fumonisin content (≥100 mg kg-1), while the other genotypes were almost equally split into two groups, low (≤10 mg kg-1) and medium (from 11 to 100 mg kg-1) fumonisin content. The number of infected kernels after artificial inoculation correlated with fumonisin concentration both in 2009 (r = 0.94; P≤0.01) and 2010 (r = 0.67; P≤0.01). Additionally, the percentage of internally infected kernels correlated positively with fumonisin concentration (r = 0.37; P≤0.01) and with the number of infected kernels (r = 0.29; P≤0.05). This research has demonstrated that Italian maize germplasm is a valid source of resistance to Fusarium ear rot. Furthermore, there is a strong association of visible Fusarium symptoms with fumonisin concentration, suggesting that selection in maize for reduced visible moulds should reduce the risk of mycotoxin contamination

COVID-19 in patients with thoracic malignancies (TERAVOLT): first results of an international, registry-based, cohort study

Background: Early reports on patients with cancer and COVID-19 have suggested a high mortality rate compared with the general population. Patients with thoracic malignancies are thought to be particularly susceptible to COVID-19 given their older age, smoking habits, and pre-existing cardiopulmonary comorbidities, in addition to cancer treatments. We aimed to study the effect of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection on patients with thoracic malignancies. Methods: The Thoracic Cancers International COVID-19 Collaboration (TERAVOLT) registry is a multicentre observational study composed of a cross-sectional component and a longitudinal cohort component. Eligibility criteria were the presence of any thoracic cancer (non-small-cell lung cancer [NSCLC], small-cell lung cancer, mesothelioma, thymic epithelial tumours, and other pulmonary neuroendocrine neoplasms) and a COVID-19 diagnosis, either laboratory confirmed with RT-PCR, suspected with symptoms and contacts, or radiologically suspected cases with lung imaging features consistent with COVID-19 pneumonia and symptoms. Patients of any age, sex, histology, or stage were considered eligible, including those in active treatment and clinical follow-up. Clinical data were extracted from medical records of consecutive patients from Jan 1, 2020, and will be collected until the end of pandemic declared by WHO. Data on demographics, oncological history and comorbidities, COVID-19 diagnosis, and course of illness and clinical outcomes were collected. Associations between demographic or clinical characteristics and outcomes were measured with odds ratios (ORs) with 95% CIs using univariable and multivariable logistic regression, with sex, age, smoking status, hypertension, and chronic obstructive pulmonary disease included in multivariable analysis. This is a preliminary analysis of the first 200 patients. The registry continues to accept new sites and patient data. Findings: Between March 26 and April 12, 2020, 200 patients with COVID-19 and thoracic cancers from eight countries were identified and included in the TERAVOLT registry; median age was 68·0 years (61·8-75·0) and the majority had an Eastern Cooperative Oncology Group performance status of 0-1 (142 [72%] of 196 patients), were current or former smokers (159 [81%] of 196), had non-small-cell lung cancer (151 [76%] of 200), and were on therapy at the time of COVID-19 diagnosis (147 [74%] of 199), with 112 (57%) of 197 on first-line treatment. 152 (76%) patients were hospitalised and 66 (33%) died. 13 (10%) of 134 patients who met criteria for ICU admission were admitted to ICU; the remaining 121 were hospitalised, but were not admitted to ICU. Univariable analyses revealed that being older than 65 years (OR 1·88, 95% 1·00-3·62), being a current or former smoker (4·24, 1·70-12·95), receiving treatment with chemotherapy alone (2·54, 1·09-6·11), and the presence of any comorbidities (2·65, 1·09-7·46) were associated with increased risk of death. However, in multivariable analysis, only smoking history (OR 3·18, 95% CI 1·11-9·06) was associated with increased risk of death. Interpretation: With an ongoing global pandemic of COVID-19, our data suggest high mortality and low admission to intensive care in patients with thoracic cancer. Whether mortality could be reduced with treatment in intensive care remains to be determined. With improved cancer therapeutic options, access to intensive care should be discussed in a multidisciplinary setting based on cancer specific mortality and patients' preference

Disease-Modifying Therapies and Coronavirus Disease 2019 Severity in Multiple Sclerosis

Objective: This study was undertaken to assess the impact of immunosuppressive and immunomodulatory therapies on the severity of coronavirus disease 2019 (COVID-19) in people with multiple sclerosis (PwMS). Methods: We retrospectively collected data of PwMS with suspected or confirmed COVID-19. All the patients had complete follow-up to death or recovery. Severe COVID-19 was defined by a 3-level variable: mild disease not requiring hospitalization versus pneumonia or hospitalization versus intensive care unit (ICU) admission or death. We evaluated baseline characteristics and MS therapies associated with severe COVID-19 by multivariate and propensity score (PS)-weighted ordinal logistic models. Sensitivity analyses were run to confirm the results. Results: Of 844 PwMS with suspected (n = 565) or confirmed (n = 279) COVID-19, 13 (1.54%) died; 11 of them were in a progressive MS phase, and 8 were without any therapy. Thirty-eight (4.5%) were admitted to an ICU; 99 (11.7%) had radiologically documented pneumonia; 96 (11.4%) were hospitalized. After adjusting for region, age, sex, progressive MS course, Expanded Disability Status Scale, disease duration, body mass index, comorbidities, and recent methylprednisolone use, therapy with an anti-CD20 agent (ocrelizumab or rituximab) was significantly associated (odds ratio [OR] = 2.37, 95% confidence interval [CI] = 1.18-4.74, p = 0.015) with increased risk of severe COVID-19. Recent use (<1 month) of methylprednisolone was also associated with a worse outcome (OR = 5.24, 95% CI = 2.20-12.53, p = 0.001). Results were confirmed by the PS-weighted analysis and by all the sensitivity analyses. Interpretation: This study showed an acceptable level of safety of therapies with a broad array of mechanisms of action. However, some specific elements of risk emerged. These will need to be considered while the COVID-19 pandemic persists

COVID-19 Severity in Multiple Sclerosis: Putting Data Into Context

Background and objectives: It is unclear how multiple sclerosis (MS) affects the severity of COVID-19. The aim of this study is to compare COVID-19-related outcomes collected in an Italian cohort of patients with MS with the outcomes expected in the age- and sex-matched Italian population. Methods: Hospitalization, intensive care unit (ICU) admission, and death after COVID-19 diagnosis of 1,362 patients with MS were compared with the age- and sex-matched Italian population in a retrospective observational case-cohort study with population-based control. The observed vs the expected events were compared in the whole MS cohort and in different subgroups (higher risk: Expanded Disability Status Scale [EDSS] score > 3 or at least 1 comorbidity, lower risk: EDSS score ≤ 3 and no comorbidities) by the χ2 test, and the risk excess was quantified by risk ratios (RRs). Results: The risk of severe events was about twice the risk in the age- and sex-matched Italian population: RR = 2.12 for hospitalization (p < 0.001), RR = 2.19 for ICU admission (p < 0.001), and RR = 2.43 for death (p < 0.001). The excess of risk was confined to the higher-risk group (n = 553). In lower-risk patients (n = 809), the rate of events was close to that of the Italian age- and sex-matched population (RR = 1.12 for hospitalization, RR = 1.52 for ICU admission, and RR = 1.19 for death). In the lower-risk group, an increased hospitalization risk was detected in patients on anti-CD20 (RR = 3.03, p = 0.005), whereas a decrease was detected in patients on interferon (0 observed vs 4 expected events, p = 0.04). Discussion: Overall, the MS cohort had a risk of severe events that is twice the risk than the age- and sex-matched Italian population. This excess of risk is mainly explained by the EDSS score and comorbidities, whereas a residual increase of hospitalization risk was observed in patients on anti-CD20 therapies and a decrease in people on interferon

DMTs and Covid-19 severity in MS: a pooled analysis from Italy and France

We evaluated the effect of DMTs on Covid-19 severity in patients with MS, with a pooled-analysis of two large cohorts from Italy and France. The association of baseline characteristics and DMTs with Covid-19 severity was assessed by multivariate ordinal-logistic models and pooled by a fixed-effect meta-analysis. 1066 patients with MS from Italy and 721 from France were included. In the multivariate model, anti-CD20 therapies were significantly associated (OR = 2.05, 95%CI = 1.39–3.02, p < 0.001) with Covid-19 severity, whereas interferon indicated a decreased risk (OR = 0.42, 95%CI = 0.18–0.99, p = 0.047). This pooled-analysis confirms an increased risk of severe Covid-19 in patients on anti-CD20 therapies and supports the protective role of interferon