Enzymatic hydrolysis of bacterial cellulose in the presence of a non-catalytic cerato-platanin protein

In this work, the effect of an expansin-like cerato-platanin (CP) protein as a pre-treatment for the enzymatic hydrolysis of bacterial cellulose (BC) is investigated. To this scope, cellulases from Trichoderma reesei are used as hydrolyzing agent using different enzyme/BC formulations. Turbidity experiments reveal that for the higher enzyme concentrations (formulations 0.5:1 and 1:1) the enzymatic hydrolysis of BC show similar hydrolysis kinetics and is not dependent of the CP. However, at higher BC concentrations (formulations 0.25:1 and 0.33:1), the hydrolysis of BC is hindered by the non-catalytic protein, as confirmed by the lower content of cellobiose and glucose in the presence of CP. Light scattering experiments show that the addition of CP led to an increase of the BC particle size from (445–630 nm) to (890–1.26 μm) for the formulation 1:1, which is also corroborated by atomic force microscopy and transmission electron microscopy analyses. These results suggest that CP did not positively affect the hydrolysis of BC, in contrast to what was previously observed for plant-derived cellulose. This work for the first time investigates the anomalous behavior of cerato platanin family members with regard to its loosening activity on the structure of bacterial cellulose

A low power IoT sensor node architecture for waste management within smart cities context

This paper focuses on the realization of an Internet of Things (IoT) architecture to optimize waste management in the context of Smart Cities. In particular, a novel typology of sensor node based on the use of low cost and low power components is described. This node is provided with a single-chip microcontroller, a sensor able to measure the filling level of trash bins using ultrasounds and a data transmission module based on the LoRa LPWAN (Low Power Wide Area Network) technology. Together with the node, a minimal network architecture was designed, based on a LoRa gateway, with the purpose of testing the IoT node performances. Especially, the paper analyzes in detail the node architecture, focusing on the energy saving technologies and policies, with the purpose of extending the batteries lifetime by reducing power consumption, through hardware and software optimization. Tests on sensor and radio module effectiveness are also presented

Effect of Marble Waste Powder as a Binder Replacement on the Mechanical Resistance of Cement Mortars

The quarrying of marble and its processing to produce building materials often generates a negative impact on the environment. In the Apuan Alps marble district, a renowned quarrying area in Italy since ancient times, the aquatic pollution of water bodies, caused by the presence of marble waste in the form of powder or sludge, represents a significant and current environmental problem. Depending on the different national and international regulations on waste management, the marble waste can be classified as a special non-hazardous industrial waste. If marble waste has been managed according to environmental international and national laws, it can be reused as a by-product. For this, the present work aims to evaluate the reuse of marble waste as a material in replacement for cement for producing mortars. Subsequently, the mechanical and physical tests were carried out to evaluate the specific properties of the obtained materials during and after the curing time. The results showed that replacement of cement into mortars by marble waste always causes a decrease of mechanical properties, with still acceptable values for many applications up to a substitution of less than 25%. From the collected data, the use of marble waste in the production of cement mortars represents an adequate and sustainable destination of this by-product

Effect of Marble Waste Powder as a Binder Replacement on the Mechanical Resistance of Cement Mortars

The quarrying of marble and its processing to produce building materials often generates a negative impact on the environment. In the Apuan Alps marble district, a renowned quarrying area in Italy since ancient times, the aquatic pollution of water bodies, caused by the presence of marble waste in the form of powder or sludge, represents a significant and current environmental problem. Depending on the different national and international regulations on waste management, the marble waste can be classified as a special non-hazardous industrial waste. If marble waste has been managed according to environmental international and national laws, it can be reused as a by-product. For this, the present work aims to evaluate the reuse of marble waste as a material in replacement for cement for producing mortars. Subsequently, the mechanical and physical tests were carried out to evaluate the specific properties of the obtained materials during and after the curing time. The results showed that replacement of cement into mortars by marble waste always causes a decrease of mechanical properties, with still acceptable values for many applications up to a substitution of less than 25%. From the collected data, the use of marble waste in the production of cement mortars represents an adequate and sustainable destination of this by-product

PAMP Activity of Cerato-Platanin during Plant Interaction: An -Omic Approach

Cerato-platanin (CP) is the founder of a fungal protein family consisting in non-catalytic secreted proteins, which work as virulence factors and/or as elicitors of defense responses and systemic resistance, thus acting as PAMPs (pathogen-associated molecular patterns). Moreover, CP has been defined an expansin-like protein showing the ability to weaken cellulose aggregates, like the canonical plant expansins do. Here, we deepen the knowledge on CP PAMP activity by the use of a multi-disciplinary approach: proteomic analysis, VOC (volatile organic compound) measurements, and gas exchange determination. The treatment of Arabidopsis with CP induces a differential profile either in protein expression or in VOC emission, as well changes in photosynthetic activity. In agreement with its role of defense activator, CP treatment induces down-expression of enzymes related to primary metabolism, such as RuBisCO, triosephosphate isomerase, and ATP-synthase, and reduces the photosynthesis rate. Conversely, CP increases expression of defense-related proteins and emission of some VOCs. Interestingly, CP exposure triggered the increase in enzymes involved in GSH metabolism and redox homeostasis (glutathione S-transferase, thioredoxin, Cys-peroxiredoxin, catalase) and in enzymes related to the “glucosinolate-myrosinase” system, which are the premise for synthesis of defence compounds, such as camalexin and some VOCs, respectively. The presented results are in agreement with the accepted role of CP as a PAMP and greatly increase the knowledge of plant primary defences induced by a purified fungal elicitor

Comparison of wild and domesticated hot peppers fruit: volatile emissions, pungency and protein profiles

Capsicum plant species are globally cultivated in warm and temperate regions, being important for agro­economic, biological and cultural aspects. While their worldwide spread and their ability of cross­pollination to easily hybridize play an important role in the formation of numerous species and varieties but also create confusion for their classification. For this reason, the categorization of species and varieties is complex and several methods have been used to evaluate pepper plant origin and evolution. Therefore, the objectives of this study were to compare a wild pepper (Capsicum chacoense) with other two domesticated cultivars belonging to different species such as Capsicum annuum and C. baccatum and draw conclusions about their origins using different approaches. For this purpose three methodologies have been used and compared: the comparison of their fruits volatile organic compounds (VOCs) emissions , their capsaicin and dihydrocapsaicin content and the leaves proteomic profiles. The VOCs analysis has been conducted by a time­of­flight mass spectrometry (ToF­MS) with an innovative approach to better identify all the compounds detected, in particular using two different ionization agents (H3 O+ and NO+ ) to better identify all the compounds detected. The VOCs and pungency analyses were then used to build back propagation neural networks (BPNN) and a Random Tree classifier to conduct a multivariate analysis and evaluate the most species­specific volatiles. The outcomes appeared to be a most accurate approach with respect to the traditional varieties descriptors used for peppers discrimination. The BPNN led to the identification of several putative volatiles as good candidates for the recognition of these species or significant nodes in a decision learning tool. Finally, protein profiles have been obtained by SDS­PAGE analysis on the leaves to perform a fast proteomic comparison among the species. The protein profiles showed the C. baccatum and C. cha‐ coense were more similar to the domesticated pepper C. annuum

NMR investigation of random coil regions: clues to assess the different biological activity of two cerato platanin family members

Plant pathogenic fungi secrete several non-catalytic proteins involved in various aspects of the pathogenesis process. Amongst these, cerato-platanin (CP) was first identified and characterized as a PAMP (pathogen-associated molecular pattern) [1,2]. A sequence homology search revealed a set of fungal Cys-rich secreted proteins that have been grouped in the CP family. They induce synthesis of phytoalexins, overexpression of defense-related genes, H2O2 and NO production, markers of defense activation [3]. The core member of this family, CP (from C. platani) shows a double beta barrel fold [2]. Here we present the results of a CP orthologue with 73% of similarity, cerato-populin (Pop1), produced by C. populicola. Though both CP and Pop1 are host defense inducers, Pop1 shows a slower and weaker defense induction capacity than CP [4]. The aim of the present investigation is to define the basis of the different biological activity of the two proteins at a molecular level. Pop1 15N, 13C and 1H resonances have been assigned [5]. The analysis of Pop1 structure obtained by homology modelling, in comparison with the CP NMR structure, will be presented. A detailed analysis of the NMR-derived protein dynamics (fast and slow regime) and NOE data indicated differences between the two proteins, mainly located in the random coil region. Interestingly, this region was proposed to have an important role in oligosaccharides binding and in necrosis induction of leaves’ cells [6]; therefore, the different pattern of residues’ interactions might be the leading cause of their diverse biological activity. To address this hypothesis we have performed NMR experiments in the course of a titration of both Pop1 and CP with oligosaccharides. The high dependence on chemical pesticides in Europe poses large risks to both the environmental and human health; thus, reducing the use of those chemicals in crop production is one of the major objectives in sustainable agriculture. We expect that our results, besides providing new hints on the molecular mechanisms operating in plants induced resistance, will contribute to reach the major goal of environment protection. Besides that, from a basic science perspective, this work illustrates a time saving approach for the investigation of a protein structure-function relationship when the high-resolution structure of an orthologous protein is already available. [1] Pazzagli et al., Cell Biochem Biophys., 44 (2006), 512. [2] de Oliveira et al., J. Biol. Chem., 286 (2011), 17560. [3] Comparini et al., Appl. Microbiol. Biotechnol., 84 (2009), 309. [4] Lombardi et al., Phys. Plant. 149 (2013), 408. [5] Baroni et al., Biomol NMR Assign., (2013), epub. [6] Frias et al., Mol. Plant Pathology (2013), epub

Under fungal attack on a metalliferous soil: ROS or not ROS? Insights from Silene paradoxa L. growing under copper stress

We investigated how the adaptation to metalliferous environments can influence the plant response to biotic stress. In a metallicolous and a non-metallicolous population of Silene paradoxa the induction of oxidative stress and the production of callose and volatiles were evaluated in the presence of copper and of the PAMP fungal protein cerato-platanin, separately and in combination. Our results showed incompatibility between the ordinary ROS-mediated response to fungal attack and the acquired mechanisms of preventing oxidative stress in the tolerant population. A similar situation was also demonstrated by the sensitive population growing in the presence of copper but, in this case, with a lack of certain responses, such as callose production. In addition, in terms of the joint behaviour of emitted volatiles, multivariate statistics showed that not only did the populations respond differently to the presence of copper or biotic stress, but also that the biotic and abiotic stresses interacted in different ways in the two populations. Our results demonstrated that the same incompatibility of hyperaccumulators in ROS-mediated biotic stress signals also seemed to be exhibited by the excluder metallophyte, but without the advantage of being able to rely on the elemental defence for plant protection from natural enemies

Activated Phosphoinositide 3-Kinase Delta Syndrome 1: Clinical and Immunological Data from an Italian Cohort of Patients

Activated phosphoinositide 3-kinase delta syndrome 1 (APDS-1) is a recently described inborn error of immunity caused by monoallelic gain-of-function mutations in the PIK3CD gene. We reviewed for the first time medical records and laboratory data of eight Italian APDS-1 patients. Recurrent sinopulmonary infections were the most common clinical feature at onset of disease. Seven patients presented lymphoproliferative disease, at onset or during follow-up, one of which resembled hemophagocytic lymphohistiocytosis (HLH). Genetic analysis of the PIK3CD gene revealed three novel mutations: functional testing confirmed their activating nature. In the remaining patients, the previously reported variants p.E1021K (n = 4) and p.E525A (n = 1) were identified. Six patients were started on immunoglobulin replacement treatment (IgRT). One patient successfully underwent hematopoietic stem cell transplantation (HSCT), with good chimerism and no GVHD at 21 months post-HSCT. APDS-1 is a combined immune deficiency with a wide variety of clinical manifestations and a complex immunological presentation. Besides IgRT, specific therapies targeting the PI3K delta pathway will most likely become a valid aid for the amelioration of patients' clinical management and their quality of life

Consensus on COVID-19 Vaccination in Pediatric Oncohematological Patients, on Behalf of Infectious Working Group of Italian Association of Pediatric Hematology Oncology

Vaccines represent the best tool to prevent the severity course and fatal consequences of the pandemic by the new Coronavirus 2019 infection (SARS-CoV-2). Considering the limited data on vaccination of pediatric oncohematological patients, we developed a Consensus document to support the Italian pediatric hematological oncological (AIEOP) centers in a scientifically correct communication with families and patients and to promote vaccination. The topics of the Consensus were: SARS-CoV-2 infection and disease (COVID-19) in the pediatric subjects; COVID-19 vaccines (type, schedule); who and when to vaccinate; contraindications and risk of serious adverse events; rare adverse events; third dose and vaccination after COVID-19; and other general prevention measures. Using the Delphi methodology for Consensus, 21 statements and their corresponding rationale were elaborated and discussed with the representatives of 31 centers, followed by voting. A high grade of Consensus was obtained on topics such as the potential risk of severe COVID-19 outcome in pediatric oncohematological patients, the need for vaccination as a preventative measure, the type, schedule and booster dose of vaccine, the eligibility of the patients for vaccination, and the timing, definition, and management of contraindications and serious adverse events, and other general prevention measures. All 21 of the statements were approved. This consensus document highlights that children and adolescents affected by hematological and oncological diseases are a fragile category. Vaccination plays an important role to prevent COVID-19, to permit the regular administration of chemotherapy or other treatments, to perform control visits and hospital admissions, and to prevent treatment delays