Preparation and Comparison of Hydrolase-Coated Plastics

Polypropylene and polyethylene were coated with alpha-Chymotrypsin (a-CT) or subtilisin Carlsberg (SubC) or Burkholderia cepacia lipase (lipase BC) by different immobilization procedures, such as physical adsorption and covalent linking. This latter procedure was based on the chemical functionalization of the plastic surface by oxygen gas plasma treatment. Immobilization of the enzyme was carried out by using as cross-linking agent i) glutaraldehyde (GA) or ii) N’-diisopropylcarbodiimide (DIC) and N-hydroxysuccinimide (NHS). The effects of duration of the plasma treatment and the type of the immobilization procedure on the transesterification activity of the enzyme were investigated. In general polypropylene resulted a better support than polyethylene. Moreover, a-CT showed higher transesterification activity when immobilized with GA, while for SubC, DIC and NHS were better cross-linking agents than GA. No activity was observed with these enzymes when immobilization was carried out by physical adsorption. On the contrary, lipase BC immobilized by physical adsorption was even more active than the free enzyme. Concerning thermal stability, immobilized SubC was less stable than the free enzyme. Overall, these results show that plastics endowed with biocatalytic properties could be obtained by simple immobilization protocols and that optimal immobilization conditions depend on the type of starting plastic, plasma treatment, cross-linking method, and the nature of the enzyme

Functional Lipids in Autoimmune Inflammatory Diseases

Lipids are apolar small molecules known not only as components of cell membranes but also, in recent literature, as modulators of different biological functions. Herein, we focused on the bioactive lipids that can influence the immune responses and inflammatory processes regulating vascular hyperreactivity, pain, leukocyte trafficking, and clearance. In the case of excessive pro-inflammatory lipid activity, these lipids also contribute to the transition from acute to chronic inflammation. Based on their biochemical function, these lipids can be divided into different families, including eicosanoids, specialized pro-resolving mediators, lysoglycerophospholipids, sphingolipids, and endocannabinoids. These bioactive lipids are involved in all phases of the inflammatory process and the pathophysiology of different chronic autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, type-1 diabetes, and systemic lupus erythematosus

Immobilized hydrolytic enzymes exhibit antibiofilm activity against escherichia coli at sub-lethal concentrations

The effects of two commercially available immobilized enzymes (namely the glycosidase pectinase and the protease subtilisin A) at sub-lethal concentrations were investigated in terms of their influence on biofilm genesis, on the composition of the biofilm matrix, and their antibiotic synergy against Escherichia coli biofilm, used as a model system of bacterial biofilms. The best antibiofilm performance of solid-supported hydrolases was obtained at the surface concentration of 0.022 and 0.095 U/cm2 with a reduction of 1.2 and 2.3 log CFU/biofilm for pectinase and subtilisin, respectively. At these enzyme surface concentrations, the biocatalysts affected the structural composition of the biofilm matrix, impacting biofilm thickness. Finally, the immobilized hydrolases enhanced biofilm sensitivity to a clinically relevant concentration of the antibiotic ampicillin. At the final antibiotic concentration of 0.1 mg/ml, a reduction of 2 and 3.5 log10 units in presence of 0.022 Upectinase/cm2 and 0.095 Usubtilisin/cm2 was obtained, respectively, in comparison the antibiotic alone. Immobilized pectinase and subtilisin at sub-lethal concentrations demonstrated a great potential for antibiofilm applications

The impact of career insight in the relation with social networks and career self-management: Preliminary evidences from the Italian contamination lab

Universities are developing more education initiatives to increase the entrepreneurial mindset of students to enhance the social sustainability and self-employment. Young people should work to increase their managerial and soft skills in order to face the process of innovation and change. This exploratory study identifies some features of the participants in the first edition of the contamination laboratory (CLab) of the University of Salento (Lecce, Italy) whose mission is to develop creativity, soft skills and entrepreneurial mindset. In particular, it aims to investigate the relationship between career insight, social network and career self-management in a sample of University's students during a training course organized according to the basic principles of Entrepreneurship Education. Data collection is carried out before and after the project. Results highlighted that there are significant differences before and after the course attendance in terms of personal and professional growth. These preliminary results present innovative aspects. From a theoretical point of view, the study laid the groundwork for future research in employability and entrepreneurial skills topics. About the practical implications, the study can provide some suggestions to promote and plan sustainable interventions in order to encourage young entrepreneurship and employability

α-Chymotrypsin Immobilized on a Low-Density Polyethylene Surface Successfully Weakens Escherichia coli Biofilm Formation

The protease \u3b1-chymotrypsin (\u3b1-CT) was covalently immobilized on a low-density polyethylene (LDPE) surface, providing a new non-leaching material (LDPE-\u3b1-CT) able to preserve surfaces from biofilm growth over a long working timescale. The immobilized enzyme showed a transesterification activity of 1.24 nmol/h, confirming that the immobilization protocol did not negatively affect \u3b1-CT activity. Plate count viability assays, as well as confocal laser scanner microscopy (CLSM) analysis, showed that LDPE-\u3b1-CT significantly impacts Escherichia coli biofilm formation by (i) reducing the number of adhered cells ( 1270.7 \ub1 5.0%); (ii) significantly affecting biofilm thickness ( 1281.8 \ub1 16.7%), roughness ( 1213.8 \ub1 2.8%), substratum coverage ( 1263.1 \ub1 1.8%), and surface to bio-volume ratio (+7.1 \ub1 0.2-fold); and (iii) decreasing the matrix polysaccharide bio-volume (80.2 \ub1 23.2%). Additionally, CLSM images showed a destabilized biofilm with many cells dispersing from it. Notably, biofilm stained for live and dead cells confirmed that the reduction in the biomass was achieved by a mechanism that did not affect bacterial viability, reducing the chances for the evolution of resistant strains

Fatty acid composition and fat content in milk from cows grazing in the Alpine region

The variation in the fat profile of pooled milk from cows grazing in pastures in June and July at 400\u2013700\ua0m and at 1400\u20132250\ua0m of altitude was evaluated by gas chromatography and compared with that from cows stalled in barns and fed with a diet without fresh grass. The ratios unsaturated/saturated fatty acid in milk samples were 1.33, 1.71 and 1.69 in June and 1.21, 1.69 and 1.84 in July for cows fed with prepared\ua0diet, grazing at 400\u2013700\ua0m or grazing at 1400\u20132250\ua0m, respectively. Analogously, the ratios (oleic plus stearic acid)/palmitic acid were, for the same group of cows, 0.59, 0.72 and 0.78 in June and 0.56, 0.73 and 0.81 in July. In milk from pastured cows, the percentage of oleic, vaccenic, rumenic and \u3b1-linolenic acids increased as a function of the altitude; instead, that of linoleic acid and of cis-12-octadecenoic acid decreased. The yield of fat was always highest in milk from 1400 to 2250\ua0m of altitude (up to 3.6\ua0g per 100\ua0mL). For the milk collected in July at 1400\u20132250\ua0m of altitude, it was observed a decrease in the percentage of decanoic (capric) and dodecanoic acids and an increase in pentadecanoic, stearic, arachidic and docosanoic (behenic) acids. Possible reasons for the differences observed in the milk samples were discussed

Biochemical characterization of highly stable endolysins with a powerful and broad anti-Gram-negative lytic activity in the presence of weak acids

EMBO Conference on Viruses of Microbes III: Structure and Function - from Molecules to Communities (Programme and Abstract Book)info:eu-repo/semantics/publishedVersio

Expanding the set of rhodococcal Baeyer–Villiger monooxygenases by high-throughput cloning, expression and substrate screening

To expand the available set of Baeyer–Villiger monooxygenases (BVMOs), we have created expression constructs for producing 22 Type I BVMOs that are present in the genome of Rhodococcus jostii RHA1. Each BVMO has been probed with a large panel of potential substrates. Except for testing their substrate acceptance, also the enantioselectivity of some selected BVMOs was studied. The results provide insight into the biocatalytic potential of this collection of BVMOs and expand the biocatalytic repertoire known for BVMOs. This study also sheds light on the catalytic capacity of this large set of BVMOs that is present in this specific actinomycete. Furthermore, a comparative sequence analysis revealed a new BVMO-typifying sequence motif. This motif represents a useful tool for effective future genome mining efforts.