Vibrio proteases for biomedical applications: Modulating the proteolytic secretome of v. alginolyticus and v. parahaemolyticus for improved enzymes production

Proteolytic enzymes are of great interest for biotechnological purposes, and their large-scale production, as well as the discovery of strains producing new molecules, is a relevant issue. Collagenases are employed for biomedical and pharmaceutical purposes. The high specificity of collagenase-based preparations toward the substrate strongly relies on the enzyme purity. However, the overall activity may depend on the cooperation with other proteases, the presence of which may be essential for the overall enzymatic activity, but potentially harmful for cells and tissues. Vibrios produce some of the most promising bacterial proteases (including collagenases), and their exo-proteome includes several enzymes with different substrate specificities, the production and relative abundances of which strongly depend on growth conditions. We evaluated the effects of different media compositions on the proteolytic exo-proteome of Vibrio alginolyticus and its closely relative Vibrio parahaemolyticus, in order to improve the overall proteases production, as well as the yield of the desired enzymes subset. Substantial biological responses were achieved with all media, which allowed defining culture conditions for targeted improvement of selected enzyme classes, besides giving insights in possible regulatory mechanisms. In particular, we focused our efforts on collagenases production, because of the growing biotechnological interest due to their pharmaceutical/biomedical applications

AMOA, Associazione Medici Oculisti per l'Africa, un esempio di solidariet\ue0 istituzionalizzata. La missione Zimbabwe.

AMOA (Associazione Medici Oculisti per l\u2019Africa), \ue8 un\u2019associazione di volontariato legalmente costituita, con sede a Bologna, aconfessionale che non persegue fini di lucro. L\u2019associazione, improntata al volontariato, svolge metodicamente e con continuit\ue0 attivit\ue0 di cooperazione allo sviluppo in favore delle popolazioni del Terzo Mondo ed in particolare di quelle africane. A tale scopo l\u2019associazione stabilisce con le Autorit\ue0 nazionali dei Paesi in cui opera, con le istituzioni nazionali ed internazionali operanti localmente e le organizzazioni sociali, una fattiva collaborazione nel rispetto delle sovranit\ue0 nazionali e secondo i principi della cooperazione internazionale definiti dalla Carta delle Nazioni Unite. Essa opera per mezzo di medici e paramedici volontari che offrono gratuitamente una parte del loro tempo per servire ai bisogni sanitari delle popolazioni di questi Paesi che si trovano in uno stadio di sottosviluppo sanitario in campo oculistico per mancanza di strutture, risorse economiche e umane. Tra le varie missioni effettuate possiamo menzionare quelle in Senegal, Togo, Etiopia, Madagascar, Rwanda, Cameroun, Zimbabwe. L\u2019analisi del presente lavoro verter\ue0 sull\u2019operato della\u2019associazione in Zimbabwe evidenziando: l\u2019epidemiologia delle patologie oculari della zona di intervento; il trattamento delle cause pi\uf9 frequenti di cecit\ue0: cataratta, glaucoma, malposizione palpebrale da tracoma, i traumi del bulbo oculare; lo screening delle deficienze visive nell\u2019et\ue0 scolare: ipermetropia, miopia, astigmatismo, ambliopia; la correzione della presbiopia che riduce la capacit\ue0 lavorativa dopo i 40 anni; la promozione dell\u2019educazione sanitaria nelle scuole e nei dispensari per la prevenzione primaria e secondaria delle carenze visive; l\u2019addestramento del personale scolastico locale a misurare la capacit\ue0 visiva dei bambini a partire dal periodo prescolare; l\u2019aggiornamento del personale medico alla chirurgia mediante soggiorni sia in loco che in Italia ed infine le ricadute sulle condizioni di salute e benessere dei soggetti coinvolti

Pareto optimality in multilayer network growth

We model the formation of multi-layer transportation networks as a multi-objective optimization process, where service providers compete for passengers, and the creation of routes is determined by a multi-objective cost function encoding a trade-off between efficiency and competition. The resulting model reproduces well real-world systems as diverse as airplane, train and bus networks, thus suggesting that such systems are indeed compatible with the proposed local optimization mechanisms. In the specific case of airline transportation systems, we show that the networks of routes operated by each company are placed very close to the theoretical Pareto front in the efficiency-competition plane, and that most of the largest carriers of a continent belong to the corresponding Pareto front. Our results shed light on the fundamental role played by multi-objective optimization principles in shaping the structure of large-scale multilayer transportation systems, and provide novel insights to service providers on the strategies for the smart selection of novel routes

Optimum channel allocation in OFDMA multi-cell systems

This paper addresses the problem of allocating users to radio resources (i.e., sub-carriers) in the downlink of an OFDMA cellular system. We consider a classical multi-cellular environment with a realistic interference model and a margin adaptive approach, i.e., we aim at minimizing total transmission power while maintaining a certain given rate for each user. We discuss computational complexity issues of the resulting model and present a heuristic approach that finds optima under suitable conditions, or "reasonably good" solutions in the general case. Computational experiences show that, in a comparison with a commercial state-of-the-art optimization solver, our algorithm is quite effective in terms of both infeasibilities and transmitted powers and extremely efficient in terms of CPU times. © 2009 Springer Berlin Heidelberg

Health Care Waste production: measures and estimates in “V. Cervello” Hospital, Palermo, Italy.

A monitoring work was carried out in May/June 2007 in one large hospital located in Palermo. The monitoring consisted in weighing the infectious waste containers filled in some Departments purposely chosen. As a second stage of the work a comparison was attempted between the results obtained from the waste production monitoring and the Hospital’s purchases recorded in the same time. A restricted list of purchased products out of the general one was extracted. Such list allows one to calculate approximately the mass of medical devices purchased and their composition. To these materials a reasonable change in humidity after use was attributed. It was possible in this deductive way to draw a probable amount and composition of waste materials really arising from health care activities (commonly – though not rigorously – considered all infectious), whose characters is forbidden to ascertain by direct inspection

Modularity measure of networks with overlapping communities

In this paper we introduce a non-fuzzy measure which has been designed to rank the partitions of a network's nodes into overlapping communities. Such a measure can be useful for both quantifying clusters detected by various methods and during finding the overlapping community-structure by optimization methods. The theoretical problem referring to the separation of overlapping modules is discussed, and an example for possible applications is given as well

Metallothionein gene family in the sea urchin Paracentrotus lividus: Gene structure, differential expression and phylogenetic analysis

Metallothioneins (MT) are small and cysteine-rich proteins that bind metal ions such as zinc, copper, cadmium, and nickel. In order to shed some light on MT gene structure and evolution, we cloned seven Paracentrotus lividus MT genes, comparing them to Echinodermata and Chordata genes. Moreover, we performed a phylogenetic analysis of 32 MTs from different classes of echinoderms and 13 MTs from the most ancient chordates, highlighting the relationships between them. Since MTs have multiple roles in the cells, we performed RT-qPCR and in situ hybridization experiments to understand better MT functions in sea urchin embryos. Results showed that the expression of MTs is regulated throughout development in a cell type-specific manner and in response to various metals. The MT7 transcript is expressed in all tissues, especially in the stomach and in the intestine of the larva, but it is less metal-responsive. In contrast, MT8 is ectodermic and rises only at relatively high metal doses. MT5 and MT6 expression is highly stimulated by metals in the mesenchyme cells. Our results suggest that the P. lividus MT family originated after the speciation events by gene duplications, evolving developmental and environmental sub-functionalization

An intronic cis-regulatory element is crucial for the alpha tubulin Pl-Tuba1a gene activation in the ciliary band and animal pole neurogenic domains during sea urchin development

In sea urchin development, structures derived from neurogenic territory control the swimming and feeding responses of the pluteus as well as the process of metamorphosis. We have previously isolated an alpha tubulin family member of Paracentrotus lividus (Pl-Tuba1a, formerly known as Pl-Talpha2) that is specifically expressed in the ciliary band and animal pole neurogenic domains of the sea urchin embryo. In order to identify cis-regulatory elements controlling its spatio-temporal expression, we conducted gene transfer experiments, transgene deletions and site specific mutagenesis. Thus, a genomic region of about 2.6 Kb of Pl-Tuba1a, containing four Interspecifically Conserved Regions (ICRs), was identified as responsible for proper gene expression. An enhancer role was ascribed to ICR1 and ICR2, while ICR3 exerted a pivotal role in basal expression, restricting Tuba1a expression to the proper territories of the embryo. Additionally, the mutation of the forkhead box consensus sequence binding site in ICR3 prevented Pl-Tuba1a expression

Comparison between two MBR pilot plants treating synthetic shipboard slops: the effect of salinity increase on biological performance, biomass activity and fouling tendency

The paper reports the main results of an experimental campaign carried out on two bench scale pilot plants for the treatment of synthetic shipboard slops. In particular, two membrane bioreactors (MBRs) with submerged configuration were analyzed. One MBR pilot plant (namely, Line A) was fed with synthetic shipboard slop and was subjected to a gradual increase of salinity. Conversely, the second MBR pilot plant (namely, Line B) was fed with the same synthetic shipboard slop but without salt addition, therefore operating as a \ue2\u80\u9ccontrol\ue2\u80\u9d unit. Organic carbon, hydrocarbons and ammonium removal, kinetic constants, extracellular polymeric substances (EPSs) production and membranes fouling rates have been assessed. The observed results highlighted a stress effect exerted by salinity on the biological performances, with lower removal efficiencies in the Line A compared to Line B. Significant releases of soluble EPS in Line A promoted an increase of the resistance related to particle deposition into membrane pores (pore fouling tendency), likely due to a worsening of the mixed liquor features. Such a condition enhanced the reduction of the \ue2\u80\u9cpre-filter\ue2\u80\u9d effect of the cake layer

Maintenance of a Protein Structure in the Dynamic Evolution of TIMPs over 600 Million Years

Deciphering the events leading to protein evolution represents a challenge, especially for protein families showing complex evolutionary history. Among them, TIMPs represent an ancient eukaryotic protein family widely distributed in the animal kingdom. They are known to control the turnover of the extracellular matrix and are considered to arise early during metazoan evolution, arguably tuning essential features of tissue and epithelial organization. To probe the structure and molecular evolution of TIMPs within metazoans, we report the mining and structural characterization of a large data set of TIMPs over approximately 600 Myr. The TIMPs repertoire was explored starting from the Cnidaria phylum, coeval with the origins of connective tissue, to great apes and humans. Despite dramatic sequence differences compared with highest metazoans, the ancestral proteins displayed the canonical TIMP fold. Only small structural changes, represented by an α-helix located in the N-domain, have occurred over the evolution. Both the occurrence of such secondary structure elements and the relative solvent accessibility of the corresponding residues in the three-dimensional structures raises the possibility that these sites represent unconserved element prone to accept variations