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Abstract This article defines "cultural experience" and places it in a holistic conceptual model; “the cultural city” where it plays a relevant role in improving the performing of cities. The conceptual model combines the basic elements of the heritage city, the smart city and the creative city. The city is interpreted from a threefold perspective; as a repository of resources, as a connective interface, and as the setting for citizens' life and social and professional experiences. In this context, each of these perspectives incorporates culture in a different way, enabling different models of value creation and different processes of production and reproduction of this value. In each of the urban models described above, production processes that combine symbolic, physical, financial, social, human and cultural capital in different ways and urban strategies are implemented to provide cultural experiences that ignite transformative effects through several spillovers. That means that culture, in its different dimensions, regains the role of a raw material and becomes the point of origin to activate development processes and improve urban performance. The integration of the dimensions of the heritage city, the creative city and the smart city through an enabling context is the core proposal of the “cultural city”. In alignment with the New European Agenda for Culture, we deepen the analysis in the specific spillovers on wellbeing and quality of life, citizen engagement and urban renewal as the backbone of a set of external effects of cultural experiences. In the final part of this article, we test the plausibility of this speculative proposal through some empirical evidence. We develop an OLS model with proxy indicators, that could be considered transitional indicators, for the three different potential strategies (heritage, smart, creative). The findings support the assertion that it is conceivable that the supply of cultural experiences through a variety of tactics (heritage city, smart city and creative city) can account in part for the growth of European cities in the years after the 2008 financial crisis. These strategies have contributed to the good performance of the urban device in a way that is positive, not negligible (accounting for around 50% of the variance in productivity) and statistically significant. The provision of a context that increases the cultural experiences for citizens has clearly improved the performance of European cities, and we develop some conceptual and empirical mechanisms to explain and measure the socioeconomic impacts of these processes.

This article reports on the experiences and results of the European Union-funded Horizon 2020 project TCBL which has been successful in creating a European network of Textile and Clothing Business Labs aimed at the sustainable transformation of one of the most problematic industries in both social and environmental terms. The approach followed by the project was based on the diffused creation of value by and for all stakeholders, including consumers. This, in turn, implies a systemic transformation of business models, brought about by all players in the sector engaging in the experimentation of new processes and transaction patterns. In this way, all stakeholders were able to reap the benefits of innovation, and the lever of competitive advantage shifted from price to knowledge, collaboration, and shared values. In the meantime, the COVID-19 pandemic has had a devastating effect on the business models of the luxury and fast-fashion brands for which TCBL has aimed to offer an alternative path, also loosely in line with the provisions of the European Green Deal and the United Nations 2030 Agenda. Given the results attained, a two-pronged strategy for the constitution of a sustainable post-project TCBL ecosystem is now being implemented.

Abstract Bacterial nanocellulose (BC) is a highly versatile biopolymer currently pursued as a material of choice in varied themes of biomedical and material science research fields. With the aim to extend the biotechnological applications, the genetic tractability of the BC producers within the Komagataeibacter genus and its potential as an alternative host chassis in synthetic biology have been extensively studied. However, such studies have been largely focused on the model Komagataeibacter spp. Here, we present a novel K. intermedius strain capable of utilizing glucose, and glycerol sources for biomass and BC synthesis. Genome assembly identified one bacterial cellulose synthetase (bcs) operon containing the complete gene set encoding the BC biogenesis machinery (bcsI) and three additional copies (bcsII–IV). Investigations on the genetic tractability confirmed plasmid transformation, propagation of vectors with pBBR1 and p15A origin of replications and constitutive and inducible induction of recombinant protein in K. intermedius ENS15. This study provides the first report on the genetic tractability of K. intermedius, serving as starting point towards future genetic engineering of this strain.

Keypoints Different monoterpenes can be regioselectively hydroxylated by CYP153A6 monooxygenase The biotransformation with whole cells is complementary to chemical oxyfunctionalization Fed-batch biotransformations have been applied for preparative purposes

(1) Background: Vinclozolin is a popular fungicide used in fruit, ornamental plants, and vegetable crops. It has recently been seen that prolonged exposure to VZN can cause human or animal health damage to various organs, but little is known to date about its cardiovascular effects. In this study, we addressed the chronic effects of VZN on the myocardium and the enzymes involved in the cardiovascular function. (2) Methods: The animals were divided into four groups: group 1 served as the control, group 2 received 1 mg/kg of VZN by gavage, group 3 received 30 mg/kg of VZN by gavage, and group 4 received 100 mg/kg of VZN by gavage, for 30 days. (3) Results: Results showed that 100 mg/kg VZN markedly increased the plasma concentration of cardiac markers (CK-MB, cTnT, ANP, BNP). Moreover, compared to the control group, VZN treatment decreased the activity of SOD, CAT, and GPx, and downregulated the mRNA expression levels of Nrf2. Furthermore, collagen deposition was amplified owing to 100 mg/kg VZN cardiotoxicity. This harmful effect was confirmed by a histological study using hematoxylin and eosin (H&E) and Masson’s trichrome staining. (4) Conclusion: Overall, our results proved the cardiotoxicity caused by chronic exposure to VZN.

With the aim of developing bioprocesses for waste valorization and a reduced water footprint, we optimized a two-step fermentation process that employs the oleaginous yeast Cutaneotrichosporon oleaginosus for the production of oil from liquid cheese whey permeate. For the first step, the addition of urea as a cost-effective nitrogen source allowed an increase in yeast biomass production. In the second step, a syrup from candied fruit processing, another food waste supplied as carbon feeding, triggered lipid accumulation. Consequently, yeast lipids were produced at a final concentration and productivity of 38 g/L and 0.57 g/L/h respectively, which are among the highest reported values. Through this strategy, based on the valorization of liquid food wastes (WP and mango syrup) and by recovering not only nutritional compounds but also the water necessary for yeast growth and lipid production, we addressed one of the main goals of the circular economy. In addition, we set up an accurate and fast-flow cytometer method to quantify the lipid content, avoiding the extraction step and the use of solvents. This can represent an analytical improvement to screening lipids in different yeast strains and to monitoring the process at the single-cell level.

Abstract A new transaminase (VbTA) was identified from the genome of the halotolerant marine bacterium Virgibacillus 21D. Following heterologous expression in Escherichia coli, it was located entirely in the insoluble fraction. After a single mutation, identified via sequence homology analyses, the VbTA T16F mutant was successfully expressed in soluble form and characterised. VbTA T16F showed high stability towards polar organic solvents and salt exposure, accepting mainly hydrophobic aromatic amine and carbonyl substrates. The 2.0 Å resolution crystal structure of VbTA T16F is here reported, and together with computational calculations, revealed that this mutation is crucial for correct dimerisation and thus correct folding, leading to soluble protein expression.

The adaptation of sporeformers to extreme environmental conditions is frequently questioned due to their capacity to produce highly resistant endospores that are considered as resting contaminants, not representing populations adapted to the system. In this work, in order to gain a better understanding of bacterial adaptation to extreme habitats, we investigated the phenotypic and genomic characteristics of the halophile Virgibacillus sp. 21D isolated from the seawater-brine interface (SBI) of the MgCl2-saturated deep hypersaline anoxic basin Discovery located in the Eastern Mediterranean Sea. Vegetative cells of strain 21D showed the ability to grow in the presence of high concentrations of MgCl2, such as 14.28% corresponding to 1.5 M. Biolog phenotype MicroArray (PM) was adopted to investigate the strain phenotype, with reference to carbon energy utilization and osmotic tolerance. The strain was able to metabolize only 8.4% of 190 carbon sources provided in the PM1 and PM2 plates, mainly carbohydrates, in accordance with the low availability of nutrients in its habitat of origin. By using in silico DNA-DNA hybridization the analysis of strain 21D genome, assembled in one circular contig, revealed that the strain belongs to the species Virgibacillus dokdonensis. The genome presented compatible solute-based osmoadaptation traits, including genes encoding for osmotically activated glycine-betaine/carnitine/choline ABC transporters, as well as ectoine synthase enzymes. Osmoadaptation of the strain was then confirmed with phenotypic assays by using the osmolyte PM9 Biolog plate and growth experiments. Furthermore, the neutral isoelectric point of the reconstructed proteome suggested that the strain osmoadaptation was mainly mediated by compatible solutes. The presence of genes involved in iron acquisition and metabolism indicated that osmoadaptation was tailored to the iron-depleted saline waters of the Discovery SBI. Overall, both phenomics and genomics highlighted the potential capability of V. dokdonensis 21D vegetative cells to adapt to the environmental conditions in Discovery SBI.

Biocatalysis offers an alternative approach to conventional chemical processes for the production of single-isomer chiral drugs. Lipases are one of the most used enzymes in the synthesis of enantiomerically pure intermediates. The use of this type of enzyme is mainly due to the characteristics of their regio-, chemo- and enantioselectivity in the resolution process of racemates, without the use of cofactors. Moreover, this class of enzymes has generally excellent stability in the presence of organic solvents, facilitating the solubility of the organic substrate to be modified. Further improvements and new applications have been achieved in the syntheses of biologically active compounds catalyzed by lipases. This review critically reports and discusses examples from recent literature (2007 to mid-2015), concerning the synthesis of enantiomerically pure active pharmaceutical ingredients (APIs) and their intermediates in which the key step involves the action of a lipase.

A flow-based chemo-enzymatic synthesis of selected APIs (i.e., butacaine, procaine and procainamide) has been developed. A bioreactor made of MsAcT, a versatile acyltransferase from Mycobacterium smegmatis, immobilised on glyoxyl–garose, was exploited to efficiently prepare amide and ester intermediates in gram scale. Immobilised MsAcT was employed in pure organic solvent, demonstrating high stability and reusability. In-line purification of the key intermediates using polymer-bound sulphonyl chloride was added after the bioreactor, enhancing the automation of the process. A final hydrogenation step using the H-Cube reactor was further carried out to obtain the selected APIs in excellent yields (>99%), making the process fast, safe and easily handled.

Abstract Bacteriocins are peptides produced by various species of bacteria, especially lactic acid bacteria, which exhibit a large spectrum of action against spoilage bacteria and foodborne pathogens. Successful application of techniques for quantitative or qualitative bacteriocin determination relies not only on the sensitivity of the test-microorganisms, but also on the agar-medium employed. Cell free supernatants are routinely used to preliminary screen for antimicrobial activity of bacteria by means of the agar well diffusion method, but the supernatant may also include other molecules (such as medium components and/or intracellular compounds) accidentally released during cell free supernatant preparation, which may interfere with the assay. Reproducibility of bacteriocin activity against the same test-microorganisms is an important factor to be considered. Unfortunately, no specific information about bioassays standardization to determine bacteriocin activity is available in the literature. In this work, growth inhibition by means of the agar well diffusion assays were carried out on different agar-media showing a strong dependence on the agar-medium used, indicating that the inhibitory effects could also depend on the diffusion of exudates that are included in the cell-free supernatant. The results presented in this communication show that selection of the agar-medium is crucial for the bioassay response.

PURPOSEWe aimed to develop a predictive model for lung water content using ultrashort echo time (UTE) magnetic resonance imaging (MRI) and a sponge phantom. MATERIALS AND METHODSImage quality was preliminarily optimized, and the signal-to-noise ratio (SNR) of UTE was compared with that obtained from a three-dimensional fast gradient echo (FGRE) sequence. Four predetermined volumes of water (3.5, 3.0, 2.5, and 2.0 mL) were soaked in cellulose foam sponges 1.8 cm3 in size and were imaged with UTE-MRI at 1.5 and 3.0 Tesla (T). A multiple echo time experiment (range, 0.1–9.6 ms) was conducted, and the T2 signal decay curve was determined at each volume of water. A three-parameter equation was fitted to the measured signal, allowing for the calculation of proton density and T2*. The calculation error of proton density was determined as a function of echo time. The constants that allowed for the determination of unknown volumes of water from the measured proton density were calculated using linear regression. RESULTSUTE-MRI provided excellent image quality for the four phantoms and showed a higher SNR, compared to that of FGRE. Proton density decreased proportionally with the decreases in both lung water and field strength (from 3.5 to 2.0 mL; proton density range at 1.5 T, 30.5–17.3; at 3.0 T, 84.2–41.5). Minimum echo time less than 0.6 ms at 1.5 T and 1 ms at 3.0 T maintained calculation errors for proton density within the range of 0%–10%. The slopes of the lines for determining the unknown volumes of water with UTE-MRI were 0.12±0.003 at 1.5 T and 0.05±0.002 at 3.0 T (P < 0.0001). CONCLUSIONIn a sponge phantom imaged at 1.5 and 3.0 T, unknown volumes of water can be predicted with high accuracy using UTE-MRI.

Hydrolytic enantioselective cleavage of different racemic non-steroidal anti-inflammatory drugs (NSAIDs) ester derivatives has been studied. An engineered esterase form Bacillus subtilis (BS2m) significantly outperformed homologous enzymes from Halomonas elongata (HeE) and Bacillus coagulants (BCE) in the enantioselective hydrolysis of naproxen esters. Structural analysis of the three active sites highlighted key differences which explained the substrate preference. Immobilization of a chimeric BS2m-T4 lysozyme fusion (BS2mT4L1) was improved by resin screening achieving twice the recovered activity (22.1 ± 5 U/g) with respect to what had been previously reported, and was utilized in a packed bed reactor. Continuous hydrolysis of α-methyl benzene acetic acid butyl ester as a model substrate was easily achieved, albeit at low concentration (1 mM). However, the high degree of insolubility of the naproxen butyl ester resulted in a slurry which could not be efficiently bioconverted, despite the addition of co-solvents and lower substrate concentration (1 mM). Addition of Triton® X-100 to the substrate mix yielded 24% molar conversion and 80% e.e. at a 5 mM scale with 5 min residence time and sufficient retention of catalytic efficiency after 6 h of use.

Enantiomerically enriched 2-hydroxymethylalkanoic acids were prepared by oxidative desymmetrisation of achiral 1,3-diols using immobilized cells of Acetobacter aceti in water at 28 °C. The biotransformations were first performed in batch mode with cells immobilized in dry alginate, furnishing the desired products with high molar conversion and reaction times ranging from 2 to 6 h. The biocatalytic process was intensified using a multiphasic flow reactor, where a segmented gas⁻liquid flow regime was applied for achieving an efficient O2-liquid transfer; the continuous flow systems allowed for high yields and high biocatalyst productivity.

Preparation of optically-pure derivatives of 2-hydroxy-2-(3-hydroxyphenyl)-2-phenylacetic acid of general structure 2 was accomplished by enzymatic hydrolysis of the correspondent esters. A screening with commercial hydrolases using the methyl ester of 2-hydroxy-2-(3-hydroxyphenyl)-2-phenylacetic acid (1a) showed that crude pig liver esterase (PLE) was the only preparation with catalytic activity. Low enantioselectivity was observed with substrates 1a⁻d, whereas PLE-catalysed hydrolysis of 1e proceeded with good enantioselectivity (E = 28), after optimization. Enhancement of the enantioselectivity was obtained by chemical re-esterification of enantiomerically enriched 2e, followed by sequential enzymatic hydrolysis with PLE. The preparation of optically-pure (S)-2e was validated on multi-milligram scale.

PURPOSEWe aimed to evaluate the validity of lung lobe weight assessment via computed tomography (CT) by comparing CT-derived and ex vivo measurements.MATERIALS AND METHODSUnenhanced CT scanning was performed in 30 consecutive patients before lobectomy for lung cancer. The CT images were analyzed using research software after allowing for lobar weight quantitation. The lobar weight estimated by CT was then compared with that measured after surgery using a precision scale (ex vivo measurement). Comparisons as well as assessment of intra- and interoperator variability were conducted using the Bland-Altman method and the coefficient of repeatability (CR). Correlations were examined using Pearson’s correlation analysis.RESULTSComparison analyses were feasible for 28 cases. The ex vivo lobe weight was 186.2±57.3 g, whereas the weights measured by the two operators by CT were 190.0±55 and 182.4±58.2 g, respectively. As compared with ex vivo weights, the CR was 36.4 for operator 1 and 50.4 for operator 2; the mean differences were 3.8 and -3.8 for operators 1 and 2, respectively. The intraoperator and interoperator CR were 20.9 and 36.6, respectively. The mean differences for the intra- and interoperator analysis were -1.5 and -7.5, respectively. The correlation was very high between CT-based and ex vivo measurements (r=0.95 and r=0.90 for operators 1 and 2, respectively; P < 0.001).CONCLUSIONEstimation of lung lobe weight by semi-automated CT analysis is sufficiently reproducible and in agreement with ex vivo measurements.

PURPOSEWe aimed to compare the intra- and interoperator variability of lobar volumetry and emphysema scores obtained by semi-automated and manual segmentation techniques in lung emphysema patients.MATERIALS AND METHODSIn two sessions held three months apart, two operators performed lobar volumetry of unenhanced chest computed tomography examinations of 47 consecutive patients with chronic obstructive pulmonary disease and lung emphysema. Both operators used the manual and semi-automated segmentation techniques. The intra- and interoperator variability of the volumes and emphysema scores obtained by semi-automated segmentation was compared with the variability obtained by manual segmentation of the five pulmonary lobes.RESULTSThe intra- and interoperator variability of the lobar volumes decreased when using semi-automated lobe segmentation (coefficients of repeatability for the first operator: right upper lobe, 147 vs. 96.3; right middle lobe, 137.7 vs. 73.4; right lower lobe, 89.2 vs. 42.4; left upper lobe, 262.2 vs. 54.8; and left lower lobe, 260.5 vs. 56.5; coefficients of repeatability for the second operator: right upper lobe, 61.4 vs. 48.1; right middle lobe, 56 vs. 46.4; right lower lobe, 26.9 vs. 16.7; left upper lobe, 61.4 vs. 27; and left lower lobe, 63.6 vs. 27.5; coefficients of reproducibility in the interoperator analysis: right upper lobe, 191.3 vs. 102.9; right middle lobe, 219.8 vs. 126.5; right lower lobe, 122.6 vs. 90.1; left upper lobe, 166.9 vs. 68.7; and left lower lobe, 168.7 vs. 71.6). The coefficients of repeatability and reproducibility of emphysema scores also decreased when using semi-automated segmentation and had ranges that varied depending on the target lobe and selected threshold of emphysema.CONCLUSIONSemi-automated segmentation reduces the intra- and interoperator variability of lobar volumetry and provides a more objective tool than manual technique for quantifying lung volumes and severity of emphysema.

PURPOSEWe aimed to examine current practice patterns of international thoracic radiologists regarding radiation dose management in adult thoracic computed tomography (CT) examinations. MATERIALS AND METHODSAn electronic questionnaire was sent to 800 members of five thoracic radiology societies in North America, Europe, Asia, and Latin America addressing radiation dose training and education, standard kVp and mAs settings for thoracic CT, dose reduction practices, clinical scenarios, and demographics. RESULTSOf the 800 radiologists, 146 responded to our survey. Nearly half (66/146, 45% [95% confidence interval, 37%–53%]) had no formal training in dose reduction, with “self-study of the literature” being the most common form of training (54/146, 37% [29%–45%]). One hundred and seventeen (80% [74%–87%]) had automatic exposure control, and 76 (65% [56%–74%]) used it in all patients. Notably, most respondents (89% [84%–94%]) used a 120 to 125 kVp standard setting, whereas none used 140 kVp. The most common average dose-length-product (DLP) value was 150 to 249 mGy.cm (75/146, 51% [43%–59%]), and 59% (51%–67%) delivered less than 250 mGy.cm in a 70 kg patient. There was a tendency towards higher DLP values with multidetector-row CT. Age, gender, and pregnancy were associated more with dose reduction than weight and clinical indication. CONCLUSIONEfforts for reducing patient radiation dose are highly prevalent among thoracic radiologists. Areas for improvement include reduction of default tube current settings, reduction of anatomical scan coverage, greater use of automatic exposure control, and eventually, reduction of current reference dose values. Our study emphasizes the need for international guidelines to foster greater conformity in dose reduction by thoracic radiologists.

One of the main factors contributing to the limited impact of eParticipation projects is the presence of a high level of social complexity that has been identified by Macintosh as one of the five challenges in the implementation of eParticipation practices. How to make sense of social complexity is still an open issue as well as the way governments can take benefit from the wealth of information that is already available on their constituencies’ collective behaviour. In this paper, we contend that the presence of a considerable variance in terms of political interests, educational level and technological skills makes it very difficult to design workable and effective systems to support participation. A modular strategy is then recommended requiring policy designers to make a step towards citizens rather than expecting the citizenry to move their content production activity onto the “official” spaces created for ad hoc participation.

This paper collects some evidence from a now completed EU-funded project, aimed at the localisation and institutionalisation of two eParticipatory tools, DEMOS-Plan and the Electronic Town Meeting, within real public administration processes. The independent or combined usage of the two tools, supported by the Living Labs approach has been tested in 18 local pilots across several regions of Europe, from Ulster (UK) to Sicily and Tuscany (Italy), from Turku Archipelago (Finland) to Voroklini (Cyprus). Selected policy domains include (among others): Spatial Planning, Socio-Economic Programming, Strategic Environmental Assessment, and Open Government. Deployment has led to a number of interesting implications for the European public authorities, such as: i) building up of a cost effective ICT platform that enables regular or occasional consultation of remotely and sparsely located citizens and stakeholders; ii) gradually migrating the whole administrative system related to spatial data infrastructure towards a full digitalisation of the “legally compulsory” exchanges between planning agencies, local stakeholders and the general public; and iii) setting the stage for these two facilities to become practically interoperable to each other and across different EU Member States.

This paper reports on the deployment of a multilingual Social Networking Platform in three Regions of Europe (Catalonia, Poitou-Charentes and Tuscany), in the context of an EU-funded Preparatory Action on eParticipation dealing with the issue of climate change and energy policy making at the level of the European Parliament. The US (“Obama”) approach and a novel (“European”) usage of social networks in political online discourses are compared. A recommendation to policy makers is that social networking can be useful whenever the topics under discussion are limited in scope, but also wide in implications, so that they require moving forward from “one-off” and “ad-hoc” participation experiments, towards the permanent coverage of “mission critical” Public Administration functions.

What makes eParticipation sustainable? This special issue of JeDEM summarises existing evidence and introduces some authoritative viewpoints on a theme that is gaining relevance and furthering discussion among theorists and practitioners alike. This Issue comprises three invited papers and six refereed articles, the results of which will be briefly presented in this note.