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Supplementary Table S1 shows nucleotide sequence of the scDb-hERG1-β1. Supplementary Table S2 reports a list of primers. Supplementary Figure S1 dot blot and Coomassie staining of different scDb-hERG1-β1 clones. Supplementary Figure S2 shows peptide ELISA assay performed using different peptides. Supplementary Figure S3 shows Figure S3 cell ELISA assay performed using different cancer cell lines. Supplementary Figure S4 shows dose-dependence curves of different cell lines treated with scDb-hERG1-β1. Supplementary Figure S5 shows cell cycle plots. Supplementary Figure S6 shows spheroid growth curves of different cancer cell lines. Supplementary Figure S7 shows microscopic appearance of spheroids of PANC-1, HCT116 and MIAPaca 2 cells. Supplementary Figure S8 shows sequence alignments

Non Linear Contrast Mode imaging of mouse kidneys 3 hours after the injection of 8mg/kg of scDb

mAbs, either mono- or bispecific (bsAb), represent one of the most successful approaches to treat many types of malignancies. However, there are certain limitations to the use of full length mAbs for clinical applications, which can be overcome by engineered antibody fragments. The aim of this study was to develop a small bsAb, in the format of a single-chain diabody (scDb), to efficiently target two proteins, the hERG1 potassium channel and the β1 subunit of integrin receptors, which specifically form a macromolecular complex in cancer cells. We provide evidence that the scDb we produced binds to the hERG1/β1 complex in cancer cells and tissues, but does not bind to the hERG1 channel in nonpathologic tissues, in particular the heart. The scDb-hERG1-β1 (i) downregulates the formation of the hERG1/β1 complex, (ii) inhibits Akt phosphorylation and HIF-1α expression, and (iii) decreases cell survival, proliferation, and migration in vitro. These effects only occur in cancer cells (either colon, pancreatic, or breast), but not in normal cells. In vivo, the scDb-hERG1-β1 shows a good pharmacokinetic profile, with a half-life of 13.5 hours and no general, cardiac, or renal toxicity when injected intravenously up to the dose of 8 mg/kg. The scDb-hERG1-β1 accumulates into subcutaneous xenografted tumors, arising from either colon or pancreatic human cancer cells, and induces a reduction of tumor growth and vascularization. Overall, the scDb-hERG1-β1 represents an innovative single-chain bispecific antibody for therapeutic applications in solid cancers that overexpress the hERG1/β1 integrin signaling complex.

mAbs, either mono- or bispecific (bsAb), represent one of the most successful approaches to treat many types of malignancies. However, there are certain limitations to the use of full length mAbs for clinical applications, which can be overcome by engineered antibody fragments. The aim of this study was to develop a small bsAb, in the format of a single-chain diabody (scDb), to efficiently target two proteins, the hERG1 potassium channel and the β1 subunit of integrin receptors, which specifically form a macromolecular complex in cancer cells. We provide evidence that the scDb we produced binds to the hERG1/β1 complex in cancer cells and tissues, but does not bind to the hERG1 channel in nonpathologic tissues, in particular the heart. The scDb-hERG1-β1 (i) downregulates the formation of the hERG1/β1 complex, (ii) inhibits Akt phosphorylation and HIF-1α expression, and (iii) decreases cell survival, proliferation, and migration in vitro. These effects only occur in cancer cells (either colon, pancreatic, or breast), but not in normal cells. In vivo, the scDb-hERG1-β1 shows a good pharmacokinetic profile, with a half-life of 13.5 hours and no general, cardiac, or renal toxicity when injected intravenously up to the dose of 8 mg/kg. The scDb-hERG1-β1 accumulates into subcutaneous xenografted tumors, arising from either colon or pancreatic human cancer cells, and induces a reduction of tumor growth and vascularization. Overall, the scDb-hERG1-β1 represents an innovative single-chain bispecific antibody for therapeutic applications in solid cancers that overexpress the hERG1/β1 integrin signaling complex.

The urgency of sustainability transition requires large incumbents in the food industry to implement sustainability-oriented innovation (SOI). However, the high concentration of the food sector and the complexity of the sustainability concept make its understanding and overall transition challenging and slow. Incumbents would need to drive the transition by redesigning business models and practices and acquiring new competencies to integrate sustainability into their innovation strategy. This paper has a twofold aim: (I) analyzing the evolution of sustainability understanding over time and (II) evaluating the extent of dynamic capabilities of food incumbents to foster SOI. We developed an integrated theoretical framework combining the theory of dynamic capabilities with aspects of SOI and applied it to the case of the Norwegian food industry. We interviewed eight food incumbents and one food industry association, and we reviewed their annual and sustainability reports from 2016 till 2020. Key findings show a high strategic activity in SOI, as well as a notable and industry-wide ambiguity about what sustainability means in the food sector. Most companies reveal both an adaptive and expanding behavior implementing conscious sustainability-integrated product and process innovations. Most innovations are incremental without a radical modification of business models. Some exceptions have been detected resembling transformative changes. Clear initiatives of moving away from a linear supply chain to a more systematic approach are currently happening through food system collaborations.The online version contains supplementary material available at 10.1007/s43615-022-00234-1.

International audience; This paper explores the strategic goals behind actors’ involvement in emerging technological innovation systems (TISs) for sustainable technologies and the way they contribute to the development of the TIS functions. Based on six expert interviews with firms within the biorefinery sector in Germany, we observe four different strategic goals driving firm-actors’ involvement: (1) exploiting new markets and businesses, (2) learning about the potential of new markets and businesses, (3) developing new technologies and (4) building up new market applications. We also differentiate between three firm-actor types: (1) a leading type of incumbent firm highly contributing to the TIS functions; (2) a learning type of incumbent firm with lower contribution; and (3) a ‘fill the gap’ type of SME or start-up producing complementary knowledge or connecting other TIS actors.

// Claudia Duranti 1 , Laura Carraresi 2 , Angelica Sette 1, 4 , Matteo Stefanini 2 , Tiziano Lottini 1 , Silvia Crescioli 1, 5 , Olivia Crociani 1 , Luisa Iamele 3 , Hugo De Jonge 3 , Ermanno Gherardi 3 and Annarosa Arcangeli 1 1 Department of Experimental and Clinical Medicine, Section of Internal Medicine, University of Florence, Florence, Italy 2 DIVAL Toscana Srl, Sesto Fiorentino, Florence, Italy 3 Department of Molecular Medicine, University of Pavia, Pavia, Italy 4 Present address: Crescendo Biologics Ltd., Babraham Research Campus, Cambridge, UK 5 Present address: Department of Dermatology, King’s College, London, UK Correspondence to: Annarosa Arcangeli, email: annarosa.arcangeli@unifi.it Keywords: cancer diagnostics; molecular imaging; hERG1 potassium channels; antibody engineering Received: August 01, 2018 Accepted: September 21, 2018 Published: October 09, 2018 ABSTRACT Modern molecular imaging techniques have greatly improved tumor detection and post-treatment follow-up of cancer patients. In this context, antibody-based imaging is rapidly becoming the gold standard, since it combines the unique specificity of antibodies with the sensitivity of the different imaging technologies. The aim of this study was to generate and characterize antibodies in single chain Fragment variable (scFv) format directed to an emerging cancer biomarker, the human ether-a-go-go-related gene-1 (hERG1) potassium channel, and to obtain a proof of concept for their potential use for in vivo molecular imaging. The anti-hERG1scFv was generated from a full length monoclonal antibody and then mutagenized, substituting a Phenylalanine residue in the third framework of the V H domain with a Cysteine residue. The resulting scFv-hERG1-Cys showed much higher stability and protein yield, increased affinity and more advantageous binding kinetics, compared to the “native” anti-hERG1scFv. The scFv-hERG1-Cys was hence chosen and characterized: it showed a good binding to the native hERG1 antigen expressed on cells, was stable in serum and displayed a fast pharmacokinetic profile once injected intravenously in nude mice. The calculated half-life was 3.1 hours and no general toxicity or cardiac toxic effects were detected. Finally, the in vivo distribution of an Alexa Fluor 750 conjugated scFv-hERG1-Cys was evaluated both in healthy and tumor-bearing nude mice, showing a good tumor-to-organ ratio, ideal for visualizing hERG1-expressing tumor masses in vivo . In conclusion, the scFv-hERG1-Cys possesses features which make it a suitable tool for application in cancer molecular imaging.

International audience; This paper aims to elucidate how companies can innovate their business model to strengthen their resilience and timely respond to current ecosystem challenges, like the emergence of new cross-industry value chains in the circular economy. The adoption of new sustainable technologies leads to the disruption of current linear settings and to the need of business model innovation. This study draws upon a case study from the circular economy rooted in by-product valorisation and presents an innovative bio-based process enabling phosphate recovery from rapeseed oil press-cakes. Incumbent actors of established (linear) value chains have been interviewed to highlight potential business model innovation. To this end, an inductive theory-building case study approach has been applied. Results reveal that a business model redesign is needed in order to build novel circular value chains, but it depends on chain actors’ capacity to adapt to change, namely to be resilient. The new value proposition needs to reach new customer targets more sensitive to sustainability issues and/or to enhance the awareness towards bio-based technological solutions into extant customers. Value creation and capture highly depend on the coordinating role of the chain actor willing to invest in the new bio-based technology, which becomes the focal company within the emergent value chain. Two scenarios are open, according to who is playing this role: if the new recovery and release process is carried out by an incumbent, the new process requires exploratory, transformative and exploitative learning together with extra investments in highly specific assets and the establishment of new cross-industry relationships to complement missing assets and knowledge about such technological breakthroughs available on the market. If a new actor enters the emerging value chain, business model innovation is mediated and moderated by a company owning both tangible and intangible resources, thus catalysing technology implementation.

Systems thinking (ST) represents an important cognitive paradigm for the transition towards a circular bio-economy, as greater awareness of the environmental impact of fossil-based products may lead to a switch to sustainable alternatives produced from secondary biomass which is not used as feed or food. However, the relationship between ST and the adoption of bio-based products, as well as the general mechanism of how ST affects environmental behavior, is not yet well-understood. The present study therefore aims to close these research gaps by conducting a survey-based experiment with a ST-motivated treatment, in which participants are asked to list as many consequences of their consumption behavior as possible (N=446 US consumers). Our findings suggest that the treatment is able to slightly activate a ST perspective, along with indirectly affecting consumer intentions to buy bio-based products by means of ST. Subsequent mediation analyses further reveal that an ecological worldview as well as variables from the norm-activation model function as mediators of the relationship between ST and purchase intention.

The receptor for the luteinizing hormone (LH-R) is aberrantly over expressed in cancers of the reproductive system. To uncover whether LH-R over expression has a causative role in cancer, we generated a transgenic (TG) mouse which overexpresses the human LH-R (hLH-R) in the female reproductive tract, under the control of the oviduct-specific glycoprotein (OGP) mouse promoter (mogp-1). The transgene was highly expressed in the uterus, ovary and liver, but only in the uterus morphological and molecular alterations (increased proliferation and trans-differentiation in the endometrial layer) were detected. A transcriptomic analysis on the uteri of young TG mice showed an up regulation of genes involved in cell cycle control and a down regulation of genes related to the immune system and the metabolism of xenobiotics. Aged TG females developed tumor masses in the uteri, which resembled an Endometrial Cancer (EC). Microarray and immunohistochemistry data indicated the deregulation of signaling pathways which are known to be altered in human ECs. The analysis of a cohort of 126 human ECs showed that LH-R overexpression is associated with early-stage tumors. Overall, our data led support to conclude that LH-R overexpression may directly contribute to trigger the neoplastic transformation of the endometrium.

The needed transition from a fossil-to a bio-based economy fosters the utilization of biological resources derived from recycled low value by-products and waste streams. Chain actors face multiple challenges associated with the adoption of novel processing technologies. Bio-based technologies might be related to high switching costs, missing downstream processing technologies, lack of quality standards, missing industry standards, emerging regulatory frameworks. Moreover, as previously unrelated actors form new relationships, value chains are moving towards new structural changes. This paper thus seeks to explore the challenges associated with the emergence of novel value chains with an exploratory case study approach that reviews a new process of recovering phosphate - a typical scarce and fundamental resource - from rapeseed oil press cakes. The contribution of our paper is thus not only to provide conclusions for this specific case, but also, and more importantly, to present a blueprint for other cases of scarce resources that require the establishment of novel supply chains to foster cascading usage of by-products. Expert interviews with different value chain actors have been conducted to investigate arising challenges, and derive implications and a further research agenda. Despite the advantages of the bio-based process, interviews revealed several challenges associated with the emergence of novel value chains: hesitation of respondents in making new investments in terms of equipment and know-how, missing complementary competencies, and difficulties in integrating different industrial sectors to engage in cross-industry innovation, such as food processing (rapeseed oil cakes) and specialty chemicals/mineral (phosphate and its derivatives). These exploratory results open the way for some useful recommendations, enabling us to propose an agenda for further research and policy development in areas such as: development of synergies between academia and industry; formulation of management strategies to support the flexibility of firms to capture opportunities associated with by-product valorization; building a systemic approach at a local level to implement bio-based technology clusters.

The aim of this paper is to explore consumers' perception of health-related borderline products between food and pharmaceuticals. Such products emerge due to blurring of boundaries between the food and pharmaceutical industries. They fit the description of both food and pharmaceuticals to a certain extent. Literature suggests that such products present innovation avenues to companies. However, their ultimate success largely depends on consumers’ perception of their safety, efficacy, appearance, and the product category to which they may belong (food or pharmaceuticals). Our study investigated consumers' perception of seven health-related borderline products (Probiotic yoghurt, phytosterol margarine, vitamin juice, garlic tablets, omega-3 eggs, vitamin C tablets, echinacea tea). To this end, we conducted an online survey of 104 consumers in Germany. Findings from statistical analysis indicated that consumers in Germany in general, and specifically young and highly educated women appeared to be sceptical of the health-benefits of health-related borderline products. They perceived these products to be generally safe to consume. However, consumption of such products was not perceived to be an easy method of staying healthy. Product appearance also seemed to influence consumers’ perception of health-related borderline products as food or pharmaceuticals.

In the last decades, protein engineering has developed particularly in biotechnology and pharmaceutical field. In particular, the engineered antibody subclass has arisen. The single chain diabody format (scDb), conjugating small size with antigen specificity, offers versatility representing a gold standard for a variety of applications, spacing from research to diagnostics and therapy. Along with such advantages, comes the challenge of optimizing their production, improving expression systems, purification procedures and stability. All such parameters are detrimental for protein production in general and above all for recombinant antibody expression, which has to be fine-tuned, choosing a proper protein-expression host and adjusting expression protocols accordingly. In the present paper, we present data regarding the production and purification of a single chain diabody directed against the macromolecular complex hERG1/β1 integrin. We focus on the expression of clones deriving from the transformation of Pichia pastoris yeast cells. In particular, we compare two different clones arose from two separate transformation processes, demonstrating that both are suitable for proper protein expression. Moreover, we have set up an expression protocol and compared the yields obtained using two purification machines: Akta Pure and Akta Start, with a positive outcome.

A transition towards a bio-based economy is accompanied by a growing demand for biomass resources as fossil fuels need to be replaced for the more sustainable production of consumer goods, chemicals and energy. To increase the supply of renewable biomass and avoid a conflict with food production, currently underutilized by-products (i.e. leaves, stems) from horticultural production could be valorised as feedstock. The success of this approach depends on farmers' willingness to adopt novel practices like the collection and treatment of plant leaves. However, literature on factors influencing farmers' decisions to adopt novel practices aimed to foster the Bioeconomy is limited. This paper addresses this gap by exploring drivers of farmers' interest in the valorisation of by-products. To this aim, the Value-Belief-Norm theory was used and expanded by contextual factors, such as the perceived market demand for biomass and future environmental policies. A survey with German fruit and vegetable farmers (N = 96) has been carried out and data have been analysed with a Structural Equation Model. Findings suggest that the Value-Belief-Norm theory is a relevant framework for the agricultural domain to predict farmers' interest in the valorisation of horticultural by-products. Results further indicate that an internal ecological worldview is potentially relevant for farmers' perception of contextual conditions aimed to foster the Bioeconomy. These outcomes could have managerial and policy implications associated with the identification of potential lead users to trigger the diffusion of innovative sustainable practices and generally foster the Bioeconomy.

International audience; Plastic packaging is widely associated with negative environmental consequences, which is why the food industry is increasingly interested in pro-environmental packaging alternatives- such as bio-based plastic. As the market share for bio-based plastic packaging is still small, strategies to raise consumer awareness and willingness to pay (WTP) are needed. This study integrates insights from environmental psychology and behavioral economics into choice models to assess which green nudges are most effective to increase consumer WTP for bio-based packaging. We tested a total of four strategies by providing consumers with nature pictures, reflection questions, information on bio-based plastics and normative information. Using a sample of 1019 German consumers, our results indicate that the strongest effects are generated when the nudging strategy matches the characteristic of consumers' cognitive style. For example, providing nature pictures only seems to increase WTP for bio-based packaging when consumers base their decision on their emotions and intuition. On the other hand, the strategies that provide normative information and activate the reflection about environmental consequences of plastics are most effective for consumers who enjoy cognitive deliberation.

The paper aims to analyze the relationships between strategic capabilities and performance among food SMEs, identifying which capabilities play a leading role in establishing competitive advantage. Four strategic capabilities were analyzed: innovation, marketing, network, and information acquisition. We ran a Structural Equation Model involving 67 food SMEs located in Italy. The results revealed that the marketing, network, and innovation capabilities directly and positively affect performance. SMEs benefit from selling their products in the national market. The network capability plays a dual role: It has a direct positive influence on performance as well as an indirect effect on the capability to acquire information about market and supply chain agents. The acquired market- and consumer-related information is extremely valuable in enhancing the marketing capability and improving performance. The innovation capability is slightly less significant than the others in affecting performance.[EconLit citations: L11, L25, L66, Q13]

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The Na+/K+/Cl- cotransporter-1 (NKCC1) and the K+/Cl- cotransporter-2 (KCC2) set the transmembrane Cl- gradient in the brain, and are implicated in epileptogenesis. We studied the postnatal distribution of NKCC1 and KCC2 in wild-type (WT) mice, and in a mouse model of sleep-related epilepsy, carrying the mutant β2-V287L subunit of the nicotinic acetylcholine receptor (nAChR). In WT neocortex, immunohistochemistry showed a wide distribution of NKCC1 in neurons and astrocytes. At birth, KCC2 was localized in neuronal somata, whereas at subsequent stages it was mainly found in the somatodendritic compartment. The cotransporters' expression was quantified by densitometry in the transgenic strain. KCC2 expression increased during the first postnatal weeks, while the NKCC1 amount remained stable, after birth. In mice expressing β2-V287L, the KCC2 amount in layer V of prefrontal cortex (PFC) was lower than in the control littermates at postnatal day 8 (P8), with no concomitant change in NKCC1. Consistently, the GABAergic excitatory to inhibitory switch was delayed in PFC layer V of mice carrying β2-V287L. At P60, the amount of KCC2 was instead higher in mice bearing the transgene. Irrespective of genotype, NKCC1 and KCC2 were abundantly expressed in the neuropil of most thalamic nuclei since birth. However, KCC2 expression decreased by P60 in the reticular nucleus, and more so in mice expressing β2-V287L. Therefore, a complex regulatory interplay occurs between heteromeric nAChRs and KCC2 in postnatal forebrain. The pathogenetic effect of β2-V287L may depend on altered KCC2 amounts in PFC during synaptogenesis, as well as in mature thalamocortical circuits.

Ion channels regulate cell proliferation, differentiation, and migration in normal and neoplastic cells through cell-cell and cell-extracellular matrix (ECM) transmembrane receptors called integrins. K+ flux through the human ether-a-go-go-related gene 1 (hERG1) channel shapes action potential firing in excitable cells such as cardiomyocytes. Its abundance is often aberrantly high in tumors, where it modulates integrin-mediated signaling. We found that hERG1 interacted with the beta(1) integrin subunit at the plasma membrane of human cancer cells. This interaction was not detected in cardiomyocytes because of the presence of the hERG1 auxiliary subunit KCNE1 (potassium voltage-gated channel subfamily E regulatory subunit 1), which blocked the beta(1) integrin-hERG1 interaction. Although open hERG1 channels did not interact as strongly with beta(1) integrins as did closed channels, current flow through hERG1 channels was necessary to activate the integrin-dependent phosphorylation of Tyr(397) in focal adhesion kinase (FAK) in both normal and cancer cells. In immunodeficient mice, proliferation was inhibited in breast cancer cells expressing forms of hERG1 with impaired K+ flow, whereas metastasis of breast cancer cells was reduced when the hERG1/beta(1) integrin interaction was disrupted. We conclude that the interaction of beta(1) integrins with hERG1 channels in cancer cells stimulated distinct signaling pathways that depended on the conformational state of hERG1 and affected different aspects of tumor progression.

The price-making ability of food small and medium-sized enterprises (SME) is important for their market survival. Since pricing is a part of marketing activities, by developing specific marketing capabilities, firms can improve their ability to make pricing decisions. This paper aims at evaluating the relationship between marketing capabilities and a firm’s price-making ability. A survey was conducted based on a sample of 371 food SMEs in the EU. We applied an Ordinal Regression Model to evaluate the determinants of a firm’s price-making ability. The analysis results revealed a certain ability of SMEs to set prices. Market analysis and marketing strategy planning capabilities are positively related to the price-making ability of a firm, while firm size is not. Supply chain relationships play a crucial role in price-making due to the strong constraint presented by the presence of vertical competition between food SMEs and supermarkets. (L25, L66, M31). C

// Elena Lastraioli 1 , Tiziano Lottini 1 , Jessica Iorio 1 , Giancarlo Freschi 2 , Marilena Fazi 2 , Claudia Duranti 1 , Laura Carraresi 3 , Luca Messerini 1 , Antonio Taddei 2 , Maria Novella Ringressi 2 , Marianna Salemme 4 , Vincenzo Villanacci 4 , Carla Vindigni 5 , Anna Tomezzoli 6 , Roberta La Mendola 7 , Maria Bencivenga 7 , Bruno Compagnoni 8 , Mariella Chiudinelli 9 , Luca Saragoni 10 , Ilaria Manzi 11 , Giovanni De Manzoni 7 , Paolo Bechi 2 , Luca Boni 12, * , Annarosa Arcangeli 1, * 1 Department of Experimental and Clinical Medicine, University of Florence, 50134 Florence, Italy 2 Department of Surgery and Translational Medicine, University of Florence, 50134 Florence, Italy 3 DI.V.A.L Toscana Srl, 50019 Sesto Fiorentino, Italy 4 Institute of Pathology, Spedali Civili, 25123 Brescia, Italy 5 Pathology Division, Azienda Ospedaliero-Universitaria Senese, 53100 Siena, Italy 6 Pathology Division, Borgo Trento Hospital, 37134 Verona, Italy 7 Division of Surgery, University of Verona, 37134 Verona, Italy 8 Surgery Division, Esine Hospital, ASL Vallecamonica Sebino, 25040 Esine (BS), Italy 9 Pathology Division, Esine Hospital, ASL Vallecamonica Sebino, 25040 Esine (BS), Italy 10 Pathology Division, Morgagni-Pierantoni Hospital, 47121 Forli, Italy 11 Gastroenterology and Endoscopy Unit, Morgagni-Pierantoni Hospital, 47121 Forli, Italy 12 Clinical Trials Coordinating Center, Azienda Ospedaliero-Universitaria Careggi/Istituto Toscano Tumori, 50134 Florence, Italy * These authors contributed equally to this work Correspondence to: Annarosa Arcangeli, email: annarosa.arcangeli@unifi.it Keywords: hERG1, Barrett’s esophagus, adenocarcinoma progression, surveillance, optical imaging Received: March 13, 2016 Accepted: July 09, 2016 Published: August 09, 2016 ABSTRACT Barrett’s esophagus (BE) is the only well-known precursor lesion of esophageal adenocarcinoma (EA). The exact estimates of the annual progression rate from BE to EA vary from 0.07% to 3.6%. The identification of BE patients at higher risk to progress to EA is hence mandatory, although difficult to accomplish. In search of novel BE biomarkers we analyzed the efficacy of hERG1 potassium channels in predicting BE progression to EA. Once tested by immunohistochemistry (IHC) on bioptic samples, hERG1 was expressed in BE, and its expression levels increased during progression from BE to esophageal dysplasia (ED) and EA. hERG1 was also over-expressed in the metaplastic cells arising in BE lesions obtained in different BE mouse models, induced either surgically or chemically. Furthermore, transgenic mice which over express hERG1 in the whole gastrointestinal tract, developed BE lesions after an esophago-jejunal anastomosis more frequently, compared to controls. A case-control study was performed on 104 bioptic samples from newly diagnosed BE patients further followed up for at least 10 years. It emerged a statistically significant association between hERG1 expression status and risk of progression to EA. Finally, a novel fluorophore- conjugated recombinant single chain variable fragment antibody (scFv-hERG1-Alexa488) was tested on freshly collected live BE biopsies: it could recognize hERG1 positive samples, perfectly matching IHC data. Overall, hERG1 can be considered a novel BE biomarker to be exploited for a novel endoscopic surveillance protocol, either in biopsies or through endoscopy, to identify those BE patients with higher risk to progress to EA.

PurposeThe purpose of this paper is to evaluate the market orientation and the marketing management capabilities of SMEs that produce traditional food products, by identifying their critical points in marketing activities.Design/methodology/approachThe theoretical framework of this analysis relates to market orientation approach. Marketing management capability was evaluated through a questionnaire using proxies to study the different stages of marketing management. The survey involved 150 firms of which 118 are SMEs producing traditional food products, located in Hungary, Belgium, and Italy. The data were collected by a self‐administered internet survey and were analysed using multivariate analyses.FindingsCluster analysis resulted in four clusters with different marketing management capabilities. Most of the SMEs in the sample lack marketing management capabilities, even if a considerable proportion of the firms considered report good marketing capabilities that lead to a market orientation. The weakest step of market orientation is the dissemination of generated intelligence. Indeed, SMEs lack marketing organisational activities, namely in planning and implementation of marketing strategy. Moreover, SMEs with better performance check that their objectives are reached but do not compare their strategy with that of competitors. Hence, they could miss opportunities to keep up to date with market development.Originality/valueThis paper provides interesting insights about the marketing capabilities of food SMEs, which usually find it difficult to stay in the market beside large firms. Moreover, there are not many contributions in the literature about the application of market orientation approach on traditional food producers.

Small businesses often do not have a sufficient capacity to put the appropriate r&D activities into action. never- theless, they are able to be innovative towards their products, processes, distribution channels, and geographical markets. Therefore, even for small and medium-sized enterprises (SMEs), the level of innovativeness can be high. The literature shows that the firm marketing capabilities are very important for innovation in the food industry, to guarantee that innova - tion reflects the market needs. The purpose of this paper is to analyse the relationship between the level of firm innovative - ness and different steps of the marketing management process, to understand if good marketing capabilities can affect the firm innovativeness. An interactive questionnaire available on the web was used for the data collection. The survey was conducted on 468 European SMEs. Linear regression was run to assess the link between marketing activities and the level of firm innovativeness. our empirical analysis reveals that the SME marketing capabilities show significant and positive relationships with firm innovativeness.

Fabry Disease (FD) is an X-linked multisystemic lysosomal disorder caused by mutations of α-galactosidase (GLA) gene. Only a few of the 450 genetic lesions identified so far have been characterised by in vitro expression studies. Thus the significance of newly identified GLA nucleotide variants in FD patients which lead to α-galactosidase (GAL-A) amino acid substitutions or intronic changes can be uncertain. We identified three GLA mutations, c.155G > A (p.C52Y), c.548G > C (p.G183A), c.647A > G (p.Y216C) in as many individuals (two male; one female) and performed in vitro expression studies and Western blot analysis in order to clarify their functional effects. Reduced GAL-A activity and normal or partially reduced mutant proteins were present in all overexpressed mutant systems in which three-dimensional structural analysis showed that the active site was not directly involved. We hypothesize that the three new mutations affect the GAL-A protein, leading to conformational FD. When mutant proteins overexpressed in COS-1 cells and in patients' lymphocytes were tested in the presence of the 1-deoxygalactonojirimicin (DGJ) chaperone, the p.G183A and p.Y216C systems showed increased GAL-A enzyme activities and protein stabilisation while p.C52Y was not responsive. We underline that genetic, biochemical and functional studies are helpful in clarifying the consequences of the missense genetic lesions detected in FD. ERT is the elective therapy for Fabry patients, but it is not always possible to issue the enzyme's active form in all involved organs. Our study endorses the hypothesis that an active site-specific chemical chaperone, which could be administered orally, might be effective in treating GAL-A conformational defects.

The purpose of this paper is to analyse the competitive performance of EU countries for the prepared swine meat sector over the 1990–2003 period. This sector was chosen for the role traditional products play in most EU countries. To assess competitiveness, and to compare the trends over the last 15 years, the analysis evaluates several trade indices: Revealed Comparative Advantage, the Vollrath indices, Net Export Index, and Grubel-Lloyd index. The data source was the Eurostat database, considering intra-EU export and import data referred to 17 sub-sectors with eight digit codes. Moreover, cluster analysis was applied to highlight groups of countries with similar features. Good competitive performance in the prepared swine meat sector is observed in Italy, Spain, Ireland and Austria, all of which were found to be specialized and export oriented in the sector. Germany and France show positive competitive performance, but a high level of intra-industry trade and low specialization is also revealed....

Hyperacute rejection (HAR) occurring after transplantation within phylogenetically distant species is a severe reaction triggered by preexisting xenoreactive antibodies and complement activation, leading to the destruction of the donor organ. Expression of human complement inhibitors in transgenic pig organs prolongs the survival of xenograft in experimental models. Moreover, the extent of protection from hyperacute rejection is dependent on the level and site of expression of the transgenic molecules and, probably, on the combination of different molecules. In this regard a small animal model to test the efficacy of expression vectors and different human molecules could be very advantageous. A murine model developed in our laboratory was characterized by measurement of several parameters characteristic of HAR in the livers of control and transgenic mice expressing transgenic human DAF (CD55) or MCP (CD46) at the end of 2 h of perfusion with human plasma and after 1 day. The parameters studied were heamatological values of hepatic functions (GOT and GPT), induction of pro-inflammatory molecules and histopathological evaluation. Cytokines (IL-1α, IL-1β, IL-6) induction and exposure of P-selectin on the endothelial cell surface, was only observed in control animals after 2 h of perfusion, as an early event. GOT and GPT values increase drammatically after 2 h perfusion and 1 day after the treatment according to the histopathological observation of liver damage. On the contrary, the livers of hDAF or hMCP transgenic mice, under the same treatment were significantly protected although the extent of this protection is dependent on the level of expression of transgenic human molecules.

A gene named stp1+, coding for a 17.5-kDa protein, that rescues cdc25-22 when overexpressed, has been previously isolated from fission yeast. Here we describe the expression and purification of Stp1 protein as a fusion with the glutathione S-transferase in E. coli and its kinetic characterisation. Stp1 deduced protein sequence shows an high homology to members of a class of cytosolic low Mr protein phosphatase previously known to exist only in mammalian species. Stp1 has a kinetic behaviour that appears to be intermediate with respect to the two isoenzymatic forms of low Mr protein tyrosine phosphatases present in mammalian tissues. These differing kinetic characteristics are mainly due to the sequence 45–56 that is spatially close to the active site pocket.

The purpose of this paper is to evaluate the marketing management capabilities of SMEs producing traditional food products, in order to analyse the market orientation of SMEs in the food industry. Following the theoretical approach of Market Orientation, our analysis is based on an assessment of the marketing management process. The methodology refers to a survey developed through a questionnaire published on the web, and a sample of 371 firms based in Belgium, Italy, Spain, the Czech Republic and Hungary was used in the analysis. Cluster analysis was applied to find the different levels of market orientation of the firms. The results revealed a certain lack of appropriate skills in marketing management in the firms of the sample, confirming the evidence found in economic literature concerning SMEs. Nevertheless, cluster analysis outlined a group of firms with good marketing capabilities and market oriented, and these represent a great part of the sample (40%). With regard to the stages of the marketing management process, the most problematic are those of planning and implementation, and control and evaluation, highlighting the difficulties SMEs encounter in carrying out coordinated marketing; which appears to be generally characterised by poor organisational capacity.

Background Quantification studies of mutated mRNAs have not been carried out on Morquio A patients. Such studies are very important for the determination of stability of premature termination codons (PTC) bearing transcripts in order to assess the appropriateness of introducing the newly developed therapeutic strategies such as “stop codon read-through therapy”. Methods This paper focuses on the study of the GALNS gene and mRNAs in two severe forms of Morquio A patients' fibroblasts with development of a new and rapid real-time RT-PCR for detection and quantification of absolute mRNA copy number. Results We identified two new mutations c.385A > T (p.K129X) and c.899 − 1G > C) in Pt1 and a known splicing defect c.120 + 1G > A in Pt2. Using RT-PCR and real-time RT-PCR in Pt2 we detected low levels of mRNAs, suggesting its instability; in Pt1, we detected three aberrant mRNAs introducing premature stop codons, suggesting that both the c.385A > T and c.899 − 1G > C mutations produce mRNAs capable of escaping the nonsense-mediated decay (NMD) pathway. Conclusions The development of a real-time RT-PCR assay allows to absolutely quantify the GALNS mRNAs carrying mutations that lead to PTCs bearing transcripts, which escape the NMD process and are potentially suitable for the new therapeutic approach.

We have established two murine cell lines derived from Small Cell Lung Carcinomas (SCLCs) developed by HPV-E6/E7 transgenic mice. These cells named PPAP-9 and PPAP-10 were isolated from mice bearing tumors, 9 and 10 months old, respectively. The cells, 5 microm in diameter, express HPV oncoproteins and sustain tumor formation after subcutaneous injection in syngenic mice. A detailed analysis indicated the epithelial origin and the neuroendocrine differentiation of these cells. We showed by confocal immunofluorescence the expression of the epithelial marker cytokeratin 5, whose gene promoter was used to direct the expression of HPV E6/E. Cells express several neuroendocrine markers such as CGRP, MAP-2, Ash1, CgrA, Scg2. The neuroendocrine differentiation of these cells was further confirmed by electron microscopy demonstrating neuropeptides secreting granules in their cytoplasm. Furthermore, in agreement with the altered expression observed in the majority of human SCLC we showed in these cells the absence of both p53 and pRB and a dramatic reduction in the expression of Caveolin-1.

Human Papillomavirus type 16 (HPV-16) is the cause of both benign lesions and ano-genital cancers. In HPV-associated cancers the transforming properties of the expressed viral E6 and E7 proteins have been revealed by a number of different assays. We have generated transgenic mice expressing HPV-16 E6/E7 genes under the control of the murine keratin 5 gene promoter, which should confer cell-type specific expression in the basal cells of squamous stratified epithelia. Transgenic mice developed thymic hyperplasia and lung neoplasia with 100% frequency, the thymus showing a size increase at 2 months and reaching the maximum dimension at 6 months, when lung carcinomas appeared. After this time the size of hyperplastic thymi decreased, while malignant formations invaded the mediastinal area. Hepatic metastasis could be also observed in some of the animals at the autopsy and death invariably occurred around 10-11 months of age.

Small tyrosine phoshatase 1 (Stp1) is a Schizosaccharomyces pombe low-molecular-mass phosphotyrosine-phosphatase 50% identical to Saccharomyces cerevisiae Ltp1. In order to investigate the role of Stp1 in yeast, a mutant was generated having the characteristic of a dominant negative molecule. Changes in protein tyrosine phosphorylation in S. cerevisiae proteome in response to Stp1 or its dominant negative mutant expression were analyzed by high-resolution two-dimensional (2-D) electrophoresis. The most remarkable result is the modification by phosphorylation on tyrosine of several proteins involved in carbohydrate metabolism. Twelve proteins were identified on the basis of their positions in the anti-phosphotyrosine immunoblot of the 2-D electrophoresis. Ten of these present tyrosyl residues that are within the consensus sequence for protein kinase CK2 (casein kinase-2). These data open the possibility for the identification of Stp1 substrates in yeast and provide hints about the nature of tyrosine phosphorylating agents in yeast and in other organisms where bona fide tyrosine kinases are lacking.