Reduced nitric oxide levels during drought stress promote drought tolerance in barley and is associated with elevated polyamine biosynthesis

Nitric oxide (NO) is a key messenger in plant stress responses but its exact role in drought response remains unclear. To investigate the role of NO in drought response we employed transgenic barley plants (UHb) overexpressing the barley non-symbiotic hemoglobin gene HvHb1 that oxidizes NO to NO3-. Reduced NO production under drought conditions in UHb plants was associated with increased drought tolerance. Since NO biosynthesis has been related to polyamine metabolism, we investigated whether the observed drought-related NO changes could involve polyamine pathway. UHb plants showed increases in total polyamines and in particular polyamines such as spermidine. These increases correlated with the accumulation of the amino acid precursors of polyamines and with the expression of specific polyamine biosynthesis genes. This suggests a potential interplay between NO and polyamine biosynthesis during drought response. Since ethylene has been linked to NO signaling and it is also related to polyamine metabolism, we explored this connection. In vivo ethylene measurement showed that UHb plants significantly decrease ethylene production and expression of aminocyclopropane-1-carboxylic acid synthase gene, the first committed step in ethylene biosynthesis compared with wild type. These data suggest a NO-ethylene influenced regulatory node in polyamine biosynthesis linked to drought tolerance/susceptibility in barley.publishersversionPeer reviewe

The impact of direct acting antivirals on hepatitis C virus disease burden and associated costs in four European countries

Background and Aims: We assessed the clinical and economic impact of direct-acting antiviral (DAA) therapy for hepatitis C virus (HCV) in England, Italy, Romania and Spain. Methods: An HCV progression Markov model was developed considering DAA eligibility and population data during the years 2015-2019. The period of time to recover the investment in DAAs was calculated as the cost saved by avoiding estimated clinical events for 1000 standardized treated patients. A delayed treatment scenario because of coronavirus disease (COVID-19) was also developed. Results: The estimated number of avoided hepatocellular carcinoma, decompensated cirrhosis and liver transplantations over a 20-year time horizon was: 1,057 in England; 1,221 in Italy; 1,211 in Romania; and 1,103 in Spain for patients treated during 2015-2016 and 640 in England; 626 in Italy; 739 in Romania; and 643 in Spain for patients treated during 2017-2019. The cost-savings ranged from \u20ac 45 to \u20ac 275 million. The investment needed to expand access to DAAs in 2015-2019 is estimated to be recovered in 6.5 years in England; 5.4 years in Italy; 6.7 years in Romania; and 4.5 years in Spain. A delay in treatment because of COVID-19 will increase liver mortality in all countries. Conclusion: Direct-acting antivirals have significant clinical benefits and can bring substantial cost-savings over the next 20 years, reaching a Break-even point in a short period of time. When pursuing an exit strategy from strict lockdown measures for COVID-19, providing DAAs should remain high on the list of priorities in order to maintain HCV elimination efforts

Biosimilars: the paradox of sharing the same pharmacological action without full chemical identity

The use of biotech medicines is increasing, with consequent mounting expenses for National Health Systems (NHSs). Biosimilars should be considered an opportunity to improve access to care. On the other side, the general public might suspect to receive low-quality medicines to save money. Actually, no drugs with a lesser degree of pharmaceutical quality with respect to existing alternatives can be authorized on the ground of a lower price. Biosimilars can be authorized only if their quality is of the same level as that of the originator. There is no chemical identity between biosimilars and the originators: any differences in quality attributes must be justified and shown not to impact on the safety and efficacy of the biosimilar by scientific investigations including pre-approval nonclinical and/or clinical studies. The biosimilar safety profile may be different from the originator or change over time for the same product. Hence caveats limiting the widespread use of biosimilars yet exist and should be solved by education on the main biological issues of biotech medicines, and on continuous update of the rules set up by the Regulatory Authorities to assess biosimilarity and to monitor post-approval safety

Pricing and reimbursement of pharmaceuticals in Italy

Pharmaceutical pricing, Reimbursement classification, Pharmaceutical policy, Cost containment policy, Innovation, Italy, I18,

Alkaline phosphatase inhibition based electrochemical sensors for the detection of pesticides

This article presents a bioelectrochemical system for the determination of pesticides by enzyme inhibition data. Measurements are performed by electrochemically monitoring the inhibition of the catalytic activity of the enzyme alkaline phosphatase (ALP) either in the presence or in the absence of pesticides, and particularly of Malathion and of 2,4-dichlorophenoxyacetic acid (2,4-D), representatives of the general classes of organophosphorous and organochlorinated agents. ALP catalytic activity was determined by using two different analytical configurations: (a) an amperometric ALP based biosensors using 3-indoxyl phosphate as the enzyme substrate; (b) a system allowing the voltammetric determination of the electroactive products of the ALP catalyzed reactions by using two different substrates: phenyl phosphate (PP) and ascorbate-2-phosphate (A-2-P). Studies of cyclic voltammetry and amperometry were performed first to define the optimal experimental conditions of the electrochemical measurements. For the optimal experimental conditions (Tris-0.1 M HCl aqueous buffer solution at pH 8.0, incubation time of 30-60 min) a detection limit of 0.5-6 mug 1(-1) is obtained, with a response, which is linear over 1-2 decades of concentration. (C) 2004 Elsevier B.V. All rights reserved

Regulation of non-symbiotic haemoglobin expression modulate nitric oxide levels in oats as a drought tolerance response

Trabajo presentado en la XXIV Reunión de la Sociedad Española de Biología de Plantas y XVII Congreso Hispano-Luso de Biología de Plantas, celebrados en Vigo (online) del 7 al 9 de julio de 2021.Drought tolerance responses are extremely complex and involve a wide range of processes and intricate signalling. Understanding plant responses to drought is of fundamental importance for breeding drought tolerant crops. In a previous study, we observed that transgenic barley lines overexpressing the barley non-symbiotic haemoglobin gene, HvHb1, which oxides NO to NO3- were more resistant to drought. In this work, we explored whether regulation of non-symbiotic haemoglobin expression was a natural feature involving drought responses in oats. To this aim, endogenous nitric oxide (NO) production was monitored in the oat cv. Flega and Patones, previously characterized as drought tolerant and susceptible respectively (Canales et al., 2019; Sánchez-Martín 2015). Results confirmed a NO decrease in the tolerant genotype during drought stress. Expression analysis showed that this reduction was due to overexpression of the oat haemoglobin gene leading to NO scavenging. Decreased levels of nitric oxide in the tolerant cultivar were associated with low levels of ethylene as measured in vivo, and with a reduction of ACO5 expression. The involvement of nitric oxide in the oat response to drought, through the differential expression of haemoglobin genes, was validated in 13 additional oat genotypes previously characterised as drought moderately tolerant and susceptible (Sánchez-Martín et al. 2012). There was a significant correlation between the haemoglobin expression and drought symptoms, with lower expression associated with more severe symptoms. This highlights the potential of the haemoglobin gene as a candidate gene for breeding drought tolerance in oats

Accesso precoce al mercato: dalle approvazioni condizionate di EMA agli accordi negoziali particolari di AIFA

The European Medicines Agency (EMA) has adopted assessment systems and drug registration procedures to respond to the needs for rapid patient access to therapies, such as Conditional Marketing Approval (CMA) and Under Exceptional Circumstances approval (UEC). The Italian Medicine Agency (AIFA) has introduced tools for governing prescriptive behaviour in order to promote appropriateness and correct use of drugs and to manage the uncertainty of financial impact, including Managed Entry Agreements (MEAs). The MEAs are conditional access agreements to the market for new drugs aimed at managing the uncertainty on the clinical and economic effects of the drugs themselves. The aim of this study is to analyse, through the use of the information systems of the AIFA and the institutional website of the EMA, how the drugs authorised by EMA with CMA or UEC have been managed on the access side to the Italian NHS. This in order to understand how the uncertainty deriving from the authorisation processes was subsequently reflected on the reimbursement processes to be borne by the NHS. From the analyses carried out, it emerges that 64% of the drugs approved with CMA were admitted to reimbursement and 68% of those approved with the UEC procedure. Furthermore, it can be noted that approval with CMA is predictive for the application of a financial-based or outcome-based MEA (61%), with the application of a monitoring register, unlike the drugs approved UEC for which no conditional reimbursement arrangements have been applied at the patient level. Furthermore, by carrying out a comparative analysis of the time of access to market following negotiation, from the positive opinion of the CHMP to the decision published in the Official Gazette of the Italian Republic, it emerged that the drugs approved by the UEC procedure take on average more time (897 days) to arrive at the price and reimbursement (P&R) compared to CMA drugs (636 days). Considering instead only the period from the request for P&R to the determination of P&R for the same group of drugs, the process seems to be on average faster for the drugs classified as UEC than for those with CMA (329 vs. 510 days)

MONITORING REGISTRIES AT ITALIAN MEDICINES AGENCY: FOSTERING ACCESS, GUARANTEEING SUSTAINABILITY

Objectives: The AIFA (Agenzia Italiana del FarmacoItalian Medicines Agency) Monitoring Registries track the eligibility of patients and the complete flow of treatments, guaranteeing appropriateness in use of pharmaceutical products, according to approved indications. Methods: This study describes the Italian pharmaceutical context and the aims and functioning of AIFA Monitoring Registries, focusing on the applications to the Managed Entry Agreements (MEAs) and HTA approaches. Results: The AIFA Monitoring Registries System has been operational in Italy since 2005. In 2012, the system became part of the NHS Information Technology system, aiming at enhancing appropriate use of pharmaceuticals and efficiency of the administrative activity. Currently, seventy-six medicines are monitored through the system, corresponding to fifty-eight therapeutic indications; individual treatments recorded are more than 515,000, for a population of approximately 505,000 patients. For each monitored product, patients eligible for treatment are registered in the specific therapeutic indication dynamic monitoring database to collect epidemiologic and clinical data, including data on the safety profile, and ex-post information missing at first evaluation stage. Conclusions: AIFA Monitoring Registries allow the evaluation of the pharmaceuticals' performance in clinical practice and may promote innovation and quicker access to medicines at affordable prices, for the benefit of patients