Eosinophils Target Therapy for Severe Asthma: Critical Points

Asthma is a chronic and heterogeneous disease, which is defined as severe disease whenever it requires treatment with a high dose of inhaled corticosteroids plus a second controller and/or systemic corticosteroids to prevent it from becoming ‘‘uncontrolled’’ or if it remains ‘‘uncontrolled’’ despite this therapy. Severe asthma is a heterogeneous condition consisting of phenotypes such as eosinophilic asthma, which is characterized by sputum eosinophilia, associated with mild to moderate increase in blood eosinophil count, frequently adult-onset, and associated with chronic rhinosinusitis with nasal polyps in half of the cases. Eosinophilic asthma is driven by T2 inflammation, characterized, among the others, by interleukin-5 production. IL-5 plays a key role in the differentiation, survival, migration, and activation of eosinophils, and it has become an appealing therapeutic target for eosinophilic asthma. In recent years two monoclonal antibodies (mepolizumab and reslizumab) directed against IL-5 and one monoclonal antibody directed against the alpha-subunit of the IL-5 receptor (benralizumab) have been developed. All these IL-5 target drugs have been shown to reduce the number of exacerbation in patients with severe asthma selected on the basis of peripheral blood eosinophil count. There are still a number of unresolved issues related to the anti-IL5 strategy in eosinophilic asthma, which are here reviewed. These issues include the effects of such therapy on airway obstruction and asthmatic symptoms, the level of baseline eosinophils that predicts a response to treatment, the relationship between blood and airway eosinophilia, and, perhaps most importantly, how to elucidate the pathogenetic role played by eosinophils in the individual patient with severe eosinophilic asthma

BEHAVIOUR OF Aeromonas hydrophila IN SALTED SWORDFISH SAMPLES

A challenge test for Aeromonas hydrophila in salted swordfish samples was carried out. Particularly, 24 samples (250g) were experimentally contaminated, salted and stored at two different temperature regimes (fluctuating – F group - and non fluctuating – NF group – regime). The count of A. hydrophila, Enterobacteria and Lactic acid bacteria (LAB) as well as the determination of pH and aw were performed at 0, 19 43, 163, 187, 230, 320 and 368 hours whereas the temperature was monitored continuously by using 6 data-loggers. In both group, the mean concentrations of A. hydrophila did not exceed Log 3 cfu/g and decreased below the mean value of Log 1 cfu/g after 368 hours. However in the F group the A. hydrophila growth was slower and the decrease appeared slightly higher than NF group and this suggests the temperature fluctuations induces a more pronounced behaviour variability of A. hydrophila under stressing conditions

Optimized RNA Extraction and Northern Hybridization in Streptomycetes

Northern blot hybridization is a useful tool for analyzing transcript patterns. To get a picture of what really occurs in vivo, it is necessary to use a protocol allowing full protection of the RNA integrity and recovery and unbiased transfer of the entire transcripts population. Many protocols suffer from severe limitations including only partial protection of the RNA integrity and/or loss of small sized molecules. Moreover, some of them do not allow an efficient and even transfer in the entire sizes range. These difficulties become more prominent in streptomycetes, where an initial quick lysis step is difficult to obtain. We present here an optimized northern hybridization protocol to purify, fractionate, blot, and hybridize Streptomyces RNA. It is based on grinding by a high-performance laboratory ball mill, followed by prompt lysis with acid phenol-guanidinium, alkaline transfer, and hybridization to riboprobes. Use of this protocol resulted in sharp and intense hybridization signals relative to long mRNAs previously difficult to detect

An experimental evaluation of a loop versus a reference design for two-channel microarrays

Motivation: Despite theoretical arguments that socalled "loop designs" of two-channel DNA microarray experiments are more efficient, biologists keep on using "reference designs". We describe two sets of microarray experiments with RNA from two different biological systems (TPA-stimulated mammalian cells and Streptomyces coelicor). In each case, both a loop and a reference design were performed using the same RNA preparations with the aim to study their relative efficiency. Results: The results of these experiments show that (1) the loop design attains a much higher precision than the reference design, (2) multiplicative spot effects are a large source of variability, and if they are not accounted for in the mathematical model, for example by taking log-ratios or including spot-effects, then the model will perform poorly. The first result is reinforced by a simulation study. Practical recommendations are given on how simple loop designs can be extended to more realistic experimental designs and how standard statistical methods allow the experimentalist to use and interpret the results from loop designs in practice

A simulation environment for Moving Block Signalling systems

In recent years, train-based signalling systems have drawn the infrastructure managers' attention, in the attempt to improve the line capacity and to optimize traffic. The development of a Moving Block (MVB) signalling system may offer a solution to achieve these objectives. MVB signalling relies on the definition of an occupied region which moves together with the train, which is continuously updated based on the information exchanged between the vehicle and the Radio-Block-Centre (RBC). Thus, it may safely operate an increased level of traffic on railway networks. Despite its potential benefits, there is still no real implementation of this system, which would require on-site testing operations, considering several scenarios. In the attempt to enhance the overall procedure moving towards the target of "zero on-site testing", in this work a simulation tool for MVB signalling is proposed. The simulator is developed in Matlab-Simulink environment and presents a modular and expandible architecture. It includes a behavioural model of the driver and allows for the injection of disturbances to the train motion, enabling to test unusual operational scenarios, without any risk