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3. Akkermansia muciniphila : new insights into resistance to gastrointestinal stress, adhesion, and protein interaction with human mucins through optimised in vitro trials and bioinformatics tools.

Author

Vergalito, Franca, Bagnoli, Diletta, Maiuro, Lucia, Pannella, Gianfranco, Palombo, Valentino, Testa, Bruno, Coppola, Francesca, Di Marco, Roberto M. A., Tremonte, Patrizio, Lombardi, Silvia J., Iorizzo, Massimo, Coppola, Raffaele, and Succi, Mariantonietta

Abstract

According to the FAO/WHO guidelines, selection of probiotics requires the assessment of survival under gastrointestinal stress and adhesion to human epithelial cells. These attributes were evaluated on Akkermansia muciniphila ATCC BAA-835 simulating the gastrointestinal transit (GIT) immediately followed by adhesion to human intestinal cell lines (CaCo2, HT-29, and HT-29-MTX) as an alternative approach to in vitro methods performed with fresh cells in each trial. The survival rate after GIT, as determined by plate counts and fluorescent probes, was significantly higher for A. muciniphila (about 8 Log CFU/mL) than for the probiotic Lacticaseibacillus rhamnosus GG ATCC 53103 (about 3 Log CFU/mL). The use of Live/Dead assay highlighted that A. muciniphila forms cell aggregates in the gastric phase as protective mechanism, explaining its high viability in the intestine. The rate of adhesion to human cell lines was always lower for strains tested after simulated GIT than for strains that did not undergo simulated GIT. Akkermansia muciniphila exhibited significantly higher adhesion than Lbs. rhamnosus GG, particularly to the mucus-secreting HT-29-MTX cells across a range of concentrations (2–8 Log CFU/mL). Finally, the bioinformatic analysis of A. muciniphila proteome confirmed the Amuc_1434 as a potential factor in binding to the human MUC2 protein. [ABSTRACT FROM AUTHOR]

Published
2024
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4. Plant-Based Ingredients Utilized as Fat Replacers and Natural Antimicrobial Agents in Beef Burgers.

Author

Lombardi, Silvia Jane, Pannella, Gianfranco, Coppola, Francesca, Vergalito, Franca, Maiuro, Lucia, Succi, Mariantonietta, Sorrentino, Elena, Tremonte, Patrizio, and Coppola, Raffaele

Subjects
LOCUST bean gum, FAT substitutes, LISTERIA innocua, CAROB, BEEF quality
Abstract

The present study aimed to find solutions based on the use of plant-based ingredients that would improve the nutritional quality of meat products as well as ensure sensory and microbiological quality. Two fat replacers, lemon albedo (Citrus lemon) and carob seed gum (Ceratonia siliqua), were investigated by chemical analysis and panel testing to evaluate their effect on the nutritional and sensory quality of beef burgers. The antimicrobial activity of two plant extracts, from nettle (Urtica dioica) leaves and medlar (Eriobotrya japonica) seeds, was studied, evaluating the intensity of inhibitory action and the minimum inhibitory concentration against Pseudomonas spp. and Listeria innocua strains by plate test. In addition, the antioxidant activity of both extracts was evaluated. Based on the results, lemon albedo and medlar seed extracts were validated in a food model (beef burger) by a storage test and a challenge test. The storage test results highlight that medlar seed extract prevents the formation of thiobarbituric acid reactive substances (TBARSs) and ensures microbiological quality, inhibiting Enterobacteriaceae and Pseudomonas spp. Anti-Listeria efficacy was confirmed in situ by challenge test results. In conclusion, although fat replacers ensure nutritional and sensory quality, they do not satisfy microbiological quality. This study clearly demonstrates that the safety of low-fat burgers can only be achieved through the combination of appropriate fat replacers with well-selected natural antimicrobial extracts. [ABSTRACT FROM AUTHOR]

Published
2024
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10. Diversity of fungal communities on Cabernet and Aglianico grapes from vineyards located in Southern Italy.

Author

Iorizzo, Massimo, Bagnoli, Diletta, Vergalito, Franca, Testa, Bruno, Tremonte, Patrizio, Succi, Mariantonietta, Pannella, Gianfranco, Letizia, Francesco, Albanese, Gianluca, Lombardi, Silvia Jane, and Coppola, Raffaele

Subjects
FUNGAL communities, CABERNET wines, VINEYARDS, BERRIES, GRAPES, SACCHAROMYCETACEAE, SYRAH
Abstract

Grape-associated microbial community is influenced by a combination of viticultural, climatic, pedological and anthropological factors, collectively known as terroir. Therefore, grapes of the same cultivar grown in different areas can be appreciated for their distinctive biogeographic characteristics. In our previous study, we showed that the phenotypic response of Aglianico and Cabernet grapevines from Molise and Sicily regions is significantly influenced by the prevailing pedoclimatic conditions, particularly soil physical properties. However, the scale at which microbial communities differ could be important in clarifying the concept of terroir, including whether it is linked to the grape variety present in a particular vineyard. To explore this further, in the research presented here, a comparative study on the fungal communities inhabiting the berry surfaces of Cabernet and Aglianico cultivars was conducted on different vineyards located in Southern Italy (Molise, Sicily and Campania regions, the first two of which had been involved in our previous study) by using high-throughput sequencing (HTS) and multivariate data analysis. The descriptive approach through relative abundance analysis showed the most abundant phyla (Ascomycota, Basidiomycota, and Chytridiomycota), families (Cladosporiaceae, Saccotheciaceae, Pleosporaceae, Saccharomycodaceae, Sporidiobolaceae, Didymellaceae, Filobasidiaceae, Bulleribasidiaceae, and Saccharomycetaceae) and genera (Cladosporium, Aureobasidium, Alternaria, Stemphylium and Filobasidium) detected on grape berries. The multivariate data analysis performed by using different packages (phyloseq, Vegan, mixOmics, microbiomeMarker and ggplot2) highlighted that the variable "vineyard location" significantly affect the fungal community, while the variable "grape variety" has no significant effect. Thus, some taxa are found to be part of specific vineyard ecosystems rather than specific grape varieties, giving additional information on the microbial contribution to wine quality, thanks to the presence of fermentative yeasts or, conversely, to the involvement in negative or detrimental roles, due to the presence of grape-deriving fungi implied in the spoilage of wine or in grapevine pathogenesis. In this connection, the main functions of core taxa fungi, whose role in the vineyard environment is still poorly understood, are also described. [ABSTRACT FROM AUTHOR]

Published
2024
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11. The Tumor Suppressor DAB2IP Is Regulated by Cell Contact and Contributes to YAP/TAZ Inhibition in Confluent Cells

Author

Apollonio, Mattia, primary, Bellazzo, Arianna, additional, Franco, Nicoletta, additional, Lombardi, Silvia, additional, Senigagliesi, Beatrice, additional, Casalis, Loredana, additional, Parisse, Pietro, additional, Thalhammer, Agnes, additional, Baj, Gabriele, additional, De Florian Fania, Rossella, additional, Del Sal, Giannino, additional, and Collavin, Licio, additional

Published
2023
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12. Translational readthrough at F8 nonsense variants in factor VIII B domain contributes to residual expression and lowers inhibitor association

Author

Testa, M, Lombardi, S, Bernardi, F, Ferrarese, M, Belvini, D, Radossi, P, Castaman, G, Pinotti, M, Branchini, A, Testa, Maria Francesca, Lombardi, Silvia, Bernardi, Francesco, Ferrarese, Mattia, Belvini, Donata, Radossi, Paolo, Castaman, Giancarlo, Pinotti, Mirko, Branchini, Alessio, Testa, M, Lombardi, S, Bernardi, F, Ferrarese, M, Belvini, D, Radossi, P, Castaman, G, Pinotti, M, Branchini, A, Testa, Maria Francesca, Lombardi, Silvia, Bernardi, Francesco, Ferrarese, Mattia, Belvini, Donata, Radossi, Paolo, Castaman, Giancarlo, Pinotti, Mirko, and Branchini, Alessio

Abstract

In hemophilia A, F8 nonsense variants, and particularly those affecting the large factor VIII (FVIII) B domain that is dispensable for coagulant activity, display lower association with replacement therapy-related anti-FVIII inhibitory antibodies as retrieved from multiple international databases. Since null genetic conditions favor inhibitor development, we hypothesized that translational readthrough over premature termination codons (PTC) may contribute to immune tolerance by producing full-length proteins through the insertion of amino acid subset(s). To quantitatively evaluate the readthrough output in vitro, we developed a very sensitive luciferase-based system to detect very low full-length FVIII synthesis from a wide panel (n=45; ~60% patients with PTC) of F8 nonsense variants. PTC not associated with inhibitors displayed higher readthrough-driven expression levels than inhibitor-associated PTC, a novel observation. Particularly, higher levels were detected for B-domain variants (n=20) than for variants in other domains (n=25). Studies on plasma from six hemophilia A patients with PTC, integrated by expression of the corresponding nonsense and readthrough-deriving missense variants, consistently revealed higher FVIII levels for B-domain variants. Only one B-domain PTC (Arg814*) was found among the highly represented PTC not sporadically associated with inhibitors, but with the lowest proportion of inhibitor cases (4 out of 57). These original insights into the molecular genetics of hemophilia A, and particularly into genotype-phenotype relationships related with disease treatment, demonstrate that B-domain features favor PTC readthrough output. This provides a potential molecular mechanism contributing to differential PTC-associated inhibitor occurrence, with translational implications for a novel, experimentally based classification of F8 nonsense variants.

Published
2023

13. Modulation of the One Health Approach to Tackle Brucellosis in Buffaloes and Cattle in Two Italian Territories with Different Characteristics

Author

Mazzeo, Alessandra, Tremonte, Patrizio, Rossi, Nicola, Ferrara, Carlo, Mascolo, Celestina, Lombardi, Silvia Jane, Sorrentino, Elena, Mazzeo, Alessandra, Tremonte, Patrizio, Rossi, Nicola, Ferrara, Carlo, Mascolo, Celestina, Lombardi, Silvia Jane, and Sorrentino, Elena

Abstract

The new European Union animal health law and its rule concerning brucellosis in cattle and buffaloes The authors examine the latest European Union (EU) rules concerning eradication and surveillance of brucellosis and animal infectious diseases of EU concern. The Italian rules concerning brucellosis in cattle and buffaloes Italy is included in the EU co-financed compulsory eradication and surveillance programmes for brucellosis in cattle and in buffaloes in the frame of the EU and the related Italian laws, which allowed reaching the Brucellosis Free status without vaccination (former "Officially Brucellosis Free - OBF" status) in the majority of the northern and middle Italian Regions and in some middle and southern Provinces included in Regions where the infection persists. Epidemiology of brucellosis in the EU and in Italy In the Italian Province of Caserta, the highest prevalence of brucellosis in buffaloes is reported; in 2017-2021, a total of 314 outbreaks occurred, in which 39,163 heads tested positive. Here, brucellosis is threatening not only human health and the widespread buffaloes breeding but also the important satellite activities concerning the Protected Designation of Origin (P.O.D.) cheese “Mozzarella di Bufala Campana". The authors also discuss the reemerged brucellosis in cattle in the Molise Region, which despite bordering the Province of Caserta, shows different hydrographic, orographic, and breeding characteristics. In Molise, the reemerged brucellosis had a very different epidemiological course, which allowed it to limit the adoption of One Health measures. The One Health approach to tackle brucellosis in buffaloes In order to tackle brucellosis in Campania Region and in its Province of Caserta, the One Health approach has been predisposed through strict control of animal health, human health, and the environment. The adopted model could be exportable to territories having similar characteristics.

Published
2023

14. In Vitro Assessment of Bio-Functional Properties from Lactiplantibacillus plantarum Strains

Author

Letizia, Francesco, Albanese, Gianluca, Testa, Bruno, Vergalito, Franca, Bagnoli, Diletta, Di Martino, Catello, Carillo, Petronia, Verrillo, Lucia, Succi, Mariantonietta, Sorrentino, Elena, Coppola, Raffaele, Tremonte, Patrizio, Lombardi, Silvia Jane, Di Marco, Roberto, Iorizzo, Massimo, Letizia, Francesco, Albanese, Gianluca, Testa, Bruno, Vergalito, Franca, Bagnoli, Diletta, Di Martino, Catello, Carillo, Petronia, Verrillo, Lucia, Succi, Mariantonietta, Sorrentino, Elena, Coppola, Raffaele, Tremonte, Patrizio, Jane Lombardi, Silvia, Di Marco, Roberto, and Iorizzo, Massimo

Subjects
Microbiology (medical), antioxidant activity, exopolysaccharides, Lactiplantibacillus plantarum, probiotic, β-glucosidase, γ-aminobutyric acid, antioxidant activity, exopolysaccharides, Lactiplantibacillus plantarum, probiotic, β-glucosidase, γ-aminobutyric acid, General Medicine, Molecular Biology, Microbiology
Abstract

In recent years, alongside the conventional screening procedures for the evaluation of probiotics for human usage, the pharmaceutical and food industries have encouraged scientific research towards the selection of new probiotic bacterial strains with particular functional features. Therefore, this study intended to explore novel functional properties of five Lactiplantibacillus plantarum strains isolated from bee bread. Specifically, antioxidant, antimicrobial and β-glucosidase activities, exopolysaccharides (EPS) production and the ability to synthesize γ-aminobutyric acid (GABA) were evaluated. The results demonstrated that the investigated L. plantarum strains were effective in inhibiting the growth of some human opportunistic pathogens in vitro (Pseudomonas aeruginosa, Escherichia coli, Proteus mirabilis, Enterococcus faecalis and Staphylococcus aureus). Moreover, the evaluation of antioxidant and β-glucosidase activity and of EPS and GABA production, revealed a different behavior among the strains, testifying how these properties are strongly strain-dependent. This suggests that a careful selection within a given species is important in order to identify appropriate strains for specific biotechnological applications. The results highlighted that the five strains of L. plantarum are promising candidates for application as dietary supplements in the human diet and as microbial cultures in specific food productions.

Published
2022
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15. Versatility of Saccharomyces cerevisiae 41CM in the Brewery Sector: Use as a Starter for “Ale” and “Lager” Craft Beer Production

Author

Testa, Bruno, primary, Coppola, Francesca, additional, Letizia, Francesco, additional, Albanese, Gianluca, additional, Karaulli, Julian, additional, Ruci, Mamica, additional, Pistillo, Marco, additional, Germinara, Giacinto Salvatore, additional, Messia, Maria Cristina, additional, Succi, Mariantonietta, additional, Vergalito, Franca, additional, Tremonte, Patrizio, additional, Lombardi, Silvia Jane, additional, and Iorizzo, Massimo, additional

Published
2022
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19. Fungi Occurrence in Ready-to-Eat Hazelnuts (Corylus avellana) From Different Boreal Hemisphere Areas

Author

Lombardi, Silvia Jane, primary, Pannella, Gianfranco, additional, Tremonte, Patrizio, additional, Mercurio, Ida, additional, Vergalito, Franca, additional, Caturano, Costantino, additional, Maiuro, Lucia, additional, Iorizzo, Massimo, additional, Succi, Mariantonietta, additional, Sorrentino, Elena, additional, and Coppola, Raffaele, additional

Published
2022
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26. Probiotic Properties and Potentiality of Lactiplantibacillus plantarum Strains for the Biological Control of Chalkbrood Disease

Author

Iorizzo, Massimo, primary, Testa, Bruno, additional, Ganassi, Sonia, additional, Lombardi, Silvia Jane, additional, Ianiro, Mario, additional, Letizia, Francesco, additional, Succi, Mariantonietta, additional, Tremonte, Patrizio, additional, Vergalito, Franca, additional, Cozzolino, Autilia, additional, Sorrentino, Elena, additional, Petrarca, Sonia, additional, De Cristofaro, Antonio, additional, and Coppola, Raffaele, additional

Published
2021
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34. A compensatory U1snRNA partially rescues FAH splicing and protein expression in a splicing-defective mouse model of tyrosinemia type I

Author

Balestra, D, Scalet, D, Ferrarese, M, Lombardi, S, Ziliotto, N, C. Croes, C, Petersen, N, Bosma, P, Riccardi, F, Pagani, F, Pinotti, M, van de Graaf, S, Balestra, Dario, Scalet, Daniela, Ferrarese, Mattia, Lombardi, Silvia, Ziliotto, Nicole, C. Croes, Chrystal, Petersen, Naomi, Bosma, Piter, Riccardi, Federico, Pagani, Franco, Pinotti, Mirko, van de Graaf, Stan F. J., Balestra, D, Scalet, D, Ferrarese, M, Lombardi, S, Ziliotto, N, C. Croes, C, Petersen, N, Bosma, P, Riccardi, F, Pagani, F, Pinotti, M, van de Graaf, S, Balestra, Dario, Scalet, Daniela, Ferrarese, Mattia, Lombardi, Silvia, Ziliotto, Nicole, C. Croes, Chrystal, Petersen, Naomi, Bosma, Piter, Riccardi, Federico, Pagani, Franco, Pinotti, Mirko, and van de Graaf, Stan F. J.

Abstract

The elucidation of aberrant splicing mechanisms, frequently associated with disease has led to the development of RNA therapeutics based on the U1snRNA, which is involved in 5′ splice site (5′ss) recognition. Studies in cellular models have demonstrated that engineered U1snRNAs can rescue different splicing mutation types. However, the assessment of their correction potential in vivo is limited by the scarcity of animal models with the targetable splicing defects. Here, we challenged the U1snRNA in the FAH5961SB mouse model of hepatic fumarylacetoacetate hydrolase (FAH) deficiency (Hereditary Tyrosinemia type I, HT1) due to the FAH c.706G>A splicing mutation. Through minigene expression studies we selected a compensatory U1snRNA (U1F) that was able to rescue this mutation. Intriguingly, adeno-associated virus-mediated delivery of U1F (AAV8-U1F), but not of U1wt, partially rescued FAH splicing in mouse hepatocytes. Consistently, FAH protein was detectable only in the liver of AAV8-U1F treated mice, which displayed a slightly prolonged survival. Moreover, RNA sequencing revealed the negligible impact of the U1F on the splicing profile and overall gene expression, thus pointing toward gene specificity. These data provide early in vivo proof-of-principle of the correction potential of compensatory U1snRNAs in HTI and encourage further optimization on a therapeutic perspective, and translation to other splicing-defective forms of metabolic diseases.

Published
2020

38. Potential Application of Apilactobacillus kunkeei for Human Use: Evaluation of Probiotic and Functional Properties

Author

Vergalito, Franca, primary, Testa, Bruno, additional, Cozzolino, Autilia, additional, Letizia, Francesco, additional, Succi, Mariantonietta, additional, Lombardi, Silvia Jane, additional, Tremonte, Patrizio, additional, Pannella, Gianfranco, additional, Di Marco, Roberto, additional, Sorrentino, Elena, additional, Coppola, Raffaele, additional, and Iorizzo, Massimo, additional

Published
2020
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40. Antimicrobial Activity against Paenibacillus larvae and Functional Properties of Lactiplantibacillus plantarum Strains: Potential Benefits for Honeybee Health

Author

Iorizzo, Massimo, primary, Testa, Bruno, additional, Lombardi, Silvia Jane, additional, Ganassi, Sonia, additional, Ianiro, Mario, additional, Letizia, Francesco, additional, Succi, Mariantonietta, additional, Tremonte, Patrizio, additional, Vergalito, Franca, additional, Cozzolino, Autilia, additional, Sorrentino, Elena, additional, Coppola, Raffaele, additional, Petrarca, Sonia, additional, Mancini, Massimo, additional, and De Cristofaro, Antonio, additional

Published
2020
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41. Low-Fat and High-Quality Fermented Sausages

Author

Tremonte, Patrizio, primary, Pannella, Gianfranco, additional, Lombardi, Silvia Jane, additional, Iorizzo, Massimo, additional, Vergalito, Franca, additional, Cozzolino, Autilia, additional, Maiuro, Lucia, additional, Succi, Mariantonietta, additional, Sorrentino, Elena, additional, and Coppola, Raffaele, additional

Published
2020
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43. Antagonistic Activity against Ascosphaera apis and Functional Properties of Lactobacillus kunkeei Strains

Author

Iorizzo, Massimo, primary, Lombardi, Silvia Jane, additional, Ganassi, Sonia, additional, Testa, Bruno, additional, Ianiro, Mario, additional, Letizia, Francesco, additional, Succi, Mariantonietta, additional, Tremonte, Patrizio, additional, Vergalito, Franca, additional, Cozzolino, Autilia, additional, Sorrentino, Elena, additional, Coppola, Raffaele, additional, Petrarca, Sonia, additional, Mancini, Massimo, additional, and De Cristofaro, Antonio, additional

Published
2020
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44. Inoculum Strategies and Performances of Malolactic Starter Lactobacillus plantarum M10: Impact on Chemical and Sensorial Characteristics of Fiano Wine

Author

Lombardi, Silvia Jane, primary, Pannella, Gianfranco, additional, Iorizzo, Massimo, additional, Testa, Bruno, additional, Succi, Mariantonietta, additional, Tremonte, Patrizio, additional, Sorrentino, Elena, additional, Di Renzo, Massimo, additional, Strollo, Daniela, additional, and Coppola, Raffaele, additional

Published
2020
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45. A Compensatory U1snRNA Partially Rescues FAH Splicing and Protein Expression in a Splicing-Defective Mouse Model of Tyrosinemia Type I

Author

Balestra, Dario, primary, Scalet, Daniela, additional, Ferrarese, Mattia, additional, Lombardi, Silvia, additional, Ziliotto, Nicole, additional, C. Croes, Chrystal, additional, Petersen, Naomi, additional, Bosma, Piter, additional, Riccardi, Federico, additional, Pagani, Franco, additional, Pinotti, Mirko, additional, and van de Graaf, Stan F. J., additional

Published
2020
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46. Preliminary Evaluation of the Safety and Probiotic Potential of Akkermansia muciniphila DSM 22959 in Comparison with Lactobacillus rhamnosus GG

Author

Cozzolino, Autilia, primary, Vergalito, Franca, additional, Tremonte, Patrizio, additional, Iorizzo, Massimo, additional, Lombardi, Silvia J., additional, Sorrentino, Elena, additional, Luongo, Delia, additional, Coppola, Raffaele, additional, Di Marco, Roberto, additional, and Succi, Mariantonietta, additional

Published
2020
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48. Translational readthrough of GLA nonsense mutations suggests dominant-negative effects exerted by the interaction of wild-type and missense variants

Author

Lombardi, Silvia, Ferrarese, Mattia, Marchi, Saverio, Pinton, Paolo, Pinotti, Mirko, Bernardi, Francesco, and Branchini, Alessio

Abstract

Nonsense mutations are relatively frequent in the rare X-linked lysosomal α-galactosidase A (α-Gal) deficiency (Fabry disease; FD), but have been poorly investigated. Here, we evaluated the responsiveness of a wide panel (n = 14) of GLA premature termination codons (PTCs) to the RNA-based approach of drug-induced readthrough through expression of recombinant α-Gal (rGal) nonsense and missense variants. We identified four high-responders to the readthrough-inducing aminoglycoside G418 in terms of full-length protein (C56X/W209X, ≥10% of wild-type rGal) and/or activity (Q119X/W209X/Q321X, ~5-7%), resulting in normal (Q119X/Q321X) or reduced (C56X, 0.27 ± 0.11; W209X, 0.35 ± 0.1) specific activity. To provide mechanistic insights we investigated the predicted amino acid substitutions mediated by readthrough (W209C/R, C56W/R), which resulted in correct lysosomal localization and appreciable protein/activity levels for the W209C/R variants. Differently, the C56W/R variants, albeit appreciably produced and localized into lysosomes, were inactive, thus indicating detrimental effects of substitutions at this position. Noticeably, when co-expressed with the functional W209C or W209R variants, the wild-type rGal displayed a reduced specific activity (0.5 ± 0.2 and 0.6 ± 0.2, respectively) that, considering the dimeric features of the α-Gal enzyme, suggested dominant-negative effects of missense variants through their interaction with the wild-type. Overall, we provide a novel mechanism through which amino acids inserted during readthrough might impact on the functional protein output. Our findings may also have implications for the interpretation of pathological phenotypes in heterozygous FD females, and for other human disorders involving dimeric or oligomeric proteins.

Published
2019
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50. Next generation factor VIIa with enhanced half-life

Author

Ferrarese, M., Nilsen, J., Pinotti, M., Bern, M., Davidson, R. J., Camire, R. M., Lode, H. E., Roopenian, D. C., Sandlie, I., Lombardi, Silvia, Castaman, G., Andersen, J. T., and Branchini, A.

Subjects
Socio-culturale
Published
2019

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