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3. Automated curve fitting and unsupervised clustering of manganese oxide Raman responses

Author

G. Mulè, Yves Vanbrabant, and Christian Burlet

Subjects
Chemistry, Analytical chemistry, chemistry.chemical_element, 02 engineering and technology, Manganese, Overfitting, engineering.material, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, 0104 chemical sciences, symbols.namesake, Todorokite, Phase (matter), symbols, engineering, Curve fitting, General Materials Science, 0210 nano-technology, Raman spectroscopy, Biological system, Cluster analysis, Spectroscopy, Envelope (waves)
Abstract

Natural manganese oxides characterization represents a challenge due to the broad variety of their structures and geochemical compositions along with a frequent poor crystallinity. This characterization requires the ability to conduct both a phase separation and a phase association operation. In this paper, the Raman spectra acquired on a selection of natural manganese oxide minerals are first processed with an automated curve-fitting model called MnOx. The adjustment of convolution envelope is realized, thanks to the Levenberg–Marquardt algorithm applied on a set of randomly generated seed pseudo-Voigt curves. The application conditions of the automated curve-fitting and, in particular, the number of seed curves are investigated with regards to the risk of overfitting. The MnOx model outputs are in a second step treated by data mining techniques and in particular by unsupervised clustering methods. This data processing shows promising results in terms of phase separation when the number of clusters is equivalent to the number of phases. By contrast, the decrease of number of clusters leads to a phase association which reflects spectral affinities between phases. This result shows the existence of 6 to 7 vibrational bands in Mn oxides Raman spectra, with contrasting behaviours between clustering. Thereby, vibrational bands located in the low wave number domain (

Published
2017
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4. MICROBIAL COMMUNITY MODIFICATION IN DOUGH USING LEUCONOSTOC CITREUM C2.27 AS A STARTER

Author

M. Ferrara, A. Sisto, G. Mulè, P. Lavermicocca, and P. De Bellis

Subjects
dough, metagenomic analysis, fungi, food and beverages, microbial community
Abstract

Leuconostoc citreum C2.27 (ITEM 17404, http://server.ispa.cnr.it/ITEM/Collection/) was used as a starter for the preparation of focaccia, a typical Apulian flat-bread, according to a "yeast free" protocol (De Bellis et al., 2019). The microbial diversity in focaccia doughs was evaluated by plate count and culture-independent approaches using a focaccia dough with 2% (w/w) baker's yeast and without starter strain, as a control. The doughs and flours were subjected to microbiological analyses and the amount of cultivable total lactic acid bacteria (LAB), yeasts and molds was determined. Presumptive LAB and yeasts were genetically characterized by rep-PCR. Then, bacterial strains were identified by sequencing the 16S rRNA gene, while yeasts were identified by sequencing the D1/D2 domain of 26S rDNA (De Bellis et al, 2019). The experiment was repeated in triplicates. Microbiological analyses showed the absence of yeasts in the focaccia dough inoculated with the starter L. citreum C2.27. In this preparation the starter strain dominated the microbial community. The control dough showed a very high count of yeasts, identified as Saccharomyces cerevisiae, while the population of LAB was significantly lower than that observed in the dough inoculated with the starter, although characterized by a higher microbial diversity. In order to study the influence of starter strain on bacterial microbiota of focaccia dough, a metagenomic analysis has been carried out. The analysis of microbiota was performed by sequencing the V5-V6 hypervariable region of 16S rRNA gene. After quality filtering and chimera detection, reads were aligned against SILVA ribosomal RNA sequence database (https://www.arb-silva. de/) using MALT (Herbig et al., 2016). Taxonomic binning was performed with MEGAN Community Edition software v6.12 (Huson et al., 2016). The results indicated that the V5-V6 hypervariable region of 16S rRNA was suitable for studying the bacterial microbiota of doughs, highlighting a significant difference in bacterial community of focaccia dough with baker's yeast respect to dough inoculated with the starter strain. Moreover, results confirmed that L. citreum C2.27 was able to dominate the microbial environment.

Published
2019

5. Metagenomic analysis of an industrial-scale biogas plant fed with contaminated maize silage

Author

Ferrara M., V. C. Liuzzi, F. Fanelli, M. Haidukowski, M. T. Cimmarusti, E. Casaletta, A. F. Logrieco, D. H. Huson, and G. Mulè

Subjects
next generation sequencing, metagenomics, remediation, biogas, renewable energy, mycotoxin
Abstract

Biogas production represents one of the most economically attractive alternative technology for biofuel production from renewable resources. Generally, biogas plants are fed with agricultural residual products and food wastes, but the rising up of agricultural products contaminated by mycotoxins, such as maize silage not suitable for animal feeding, has pointed the question on the possibility to use this agricultural productfor biogas production. In this regards, a preliminary metagenomic analysis of microbial community residing in a mesophilic industrial-scale biogas fermenter, daily fed with contaminated maize silage, has been carried out to characterize the evolution of microbial community under the operating conditions and the mycotoxin content. Sample were collected from a biogas plant consisting of a three steps production taking place in a bioreactor, post-reactor and a storage tank. Total DNA was extracted from samples belonging to each steps of biogas production. Metagenomic analysis was carried out by analyzing the V4 variable region of bacterial and archaeal 16S rRNA gene. Mycotoxin content was analyzed in maize silage feeding the biogas plant and in the digestate from bioreactor, post-reactor and storage tank by immunoaffinity column clean-up (Myco6in1+®) and detected with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Over 3million high quality reads (about 1Gb) were generated on the Ion Torrent S5 Sequencing System. About 2.4 million reads were assigned for 16S analysis. In detail, metagenomic analysis revealed that Bacteria superkingdom was dominant (~96%) along the production steps, whereas Archaea were less represented (~4%). Within Bacteria the most abundant phylum was Firmicutes, mostly represented by Clostridia, followed by Bacteroidetes and Synergistetes. Within the superkingdom of Archaea, only microorganisms belonging to the phylum of Euryarchaeota were detected. Within Euryarchaeota the dominant genera were Methanosarcina and Methanoculleus. Chemical analysis on maize silage feeding the plants showed an initial mycotoxin contamination by DON (410 µg/kg), FB1 (3570 µg/kg), FB2 (810 µg/kg) and T-2 toxin (20 µg/kg), while AfB1, HT-2 Toxin, NIV, OTA and ZEA were not detected. After the first step of biogas production, a complete reduction of DON and T-2 content was achieved. These preliminary results suggest a possible absorption/degradation of mycotoxins in bioreactor tank and therefore further studies are needed to better elucidate the possible involvement of specific microbial taxa capable of mycotoxins reduction and the enzymatic pathways potentially involved in mycotoxin degradation.

Published
2017

6. Development of a ddRAD library preparation and HRM approaches for SNP discovery and genotyping of Italian rice cultivars Carnaroli and Roma

Author

M. Ferrara, L. Grazina, A. De Paolis, A. F. Logrieco, I. Mafra, J. S. Amaral, and G. Mulè

Subjects
next generation sequencing, authenticity, traceability, ddRAD, food, Ssingle nucleotide polymorphism
Abstract

Food authentication and traceability is a complex problem, strictly correlated to fraud and adulteration detections that dramatically affect the consumer protection. Analysis of protein, metabolite and DNA represents robust tools for food authentication. In particular, DNA-based methods are more reliable, thanks to the stability of DNA under production and processing techniques applied along the food-chain. Therefore, DNA markers offer a powerful tool to address the validation of food authenticity and traceability of primary products. Single nucleotide polymorphism (SNP) markers have become the most used markers in genetic characterization studies as well as in translational genomic even in plants. SNP are, in fact, the most abundant forms of genetic variation among individuals of a species. In particular, SNP analysis by next generation sequencing (NGS)(e.g. genotyping by sequencing (GBS) and double-digest restriction site-associated DNA sequencing (ddRAD-Seq) or by high resolution melting analysis (HRM), e.g. single-base variants and small insertions or deletions, have rapidly become popular due to their flexibility and relatively low cost. The ddRAD-Seq technology has the advantage over GBS of high accuracy read mapping by paired-end sequencing of identical loci. Progress in NGS technology has led to the availability of several plant genomes. This situation makes it possible to simulate ddRAD-Seqin silico, allowing prediction of the numbers, sizes, and genome positions of digested fragments. However, few reports have evaluated the in silico predictions by comparative experiments using several combinations of restriction enzymes and multiple samples with different SNP density. HRM analysis has several advantages over traditional methods for gene scanning and genotyping, making it faster, less laborious and more suitable for high sample throughput. In this study, two approaches are proposed for the authentication of the Italian rice cultivars Carnaroli and Roma: in silico and empirical ddRAD-Seq analysis and HRM analysis targeting an A/C SNP in exon 6, responsible for the Wxin allele. The ddRAD-Seq approach consisted of a workflow, as follows:(i) in silico prediction of optimum restriction enzymes from the reference rice genome,(ii) verification of the prediction by ddRAD-Seq data of Carnaroli and Roma genomes (iii) establishment of a computational data processing pipeline for high confidence SNP calling, and (iv) validation of SNP accuracy. In silico prediction prior to sequencing analysis will contribute to optimization of the experimental conditions for ddRAD-Seq and could help to accelerate the detection of DNA markers useful for the authentication of rice cultivars Carnaroli and Roma. Preliminary results of HRM analysis show potential for rice cultivar differentiation since Carnaroli was distinguished from Roma, among others (Carnise/Karnak, Gladio, Sant'Andrea and others) with high level of confidence (>98%). Acknowledgments: This work has been supported by the European project FOODINTEGRITY (FP7-KBBE-2013-single-stage, No 613688).

Published
2017

7. Metagenomic analysis and functional characterization of an industrial-scale biogas plant by high throughput sequencing

Author

M. Ferrara, V. C. Liuzzi, F. Fanelli, D. H. Huson, and G. Mulè

Subjects
next generation sequencing, renewable energies, biogas, Metagenomics
Abstract

Energy crisis and environmental pollution have led to an increasing interest in renewable energies. Biogas production from plant material, agricultural residual products and food wastes represents one of the most economically attractive alternative technology for biofuel production. Complex consortia of microorganisms are responsible for biomass degradation and biogas production involving several stages. Next-generation high-throughput sequencing provides a powerful tool for dissecting microbial community structure and methane-producing pathways in anaerobic digestion. A taxonomic and functional metagenomic analysis of microbial community residing in an industrial-scale biogas fermenter has been carried out at different steps of biogas production. Sample were collected from an industrial-scale mesophilic plant consisting of a three steps production taking place in a bioreactor, post-reactor and a storage tank. Total DNA was extracted from samples belonging to each steps of biogas production. Metagenomic analysis was carried out by using 16S and shotgun sequencing approach. The 16S datasets were generated by sequencing the bacterial and archaeal V4 hypervariable region. Reads from 16S sequencing were aligned against SILVA ribosomal RNA sequence database by using MALT(1), while shotgun reads were aligned against NCBI-nr protein database by using DIAMOND(2). Taxonomic binning and functional annotation were performed with MEGAN 6 software(3). About 2.9 and 11.5 million high quality reads were generated on the Ion Torrent S5 Sequencing System for 16S and shotgun approach, respectively. Metagenomic analysis revealed that the overall taxa distribution resulting from both sequencing strategies was conserved. In details, the superkingdom of Bacteria was dominant (~93%) along the production steps, whereas Archaea were less represented (~4%). Within the superkingdom of Archaea, only microorganisms belonging to the phylum of Euryarchaeota were detected. Within the key microorganisms involved in methanogenesis, data showed that during biogas production steps the abundance of Methanosarcina genus decreased from bioreactor to storage tank, with a simultaneous increase of Methanoculleus genus. Considering the key methanogenesis pathways, functional analysis supported a shift from acetotrophic methanogens to hydrogenotrophic methanogens. Results showed that the combination of both 16S and shotgun sequencing approach successfully addressed the taxonomical and functional analysis of microbial community, revealing new insights in microbial and functional dynamics during biogas production steps.

Published
2017

8. Metagenomic analysis of an industrial-scale biogas plant by high throughput sequencing

Author

M. Ferrara, V. C. Liuzzi, F. Fanelli, D. H. Huson, and G. Mulè

Subjects
next generation sequencing, renewable energies, biogas, Metagenomics
Abstract

Introduction Energy crisis and environmental pollution have led to an increasing interest in renewable energies. Biogas production from plant material, agricultural residual products and food wastes represents one of the most economically attractive alternative technology for biofuel production. In this regards, anaerobic digestion has been widely applied to produce methane for biofuel. Complex consortia of microorganisms are responsible for biomass degradation and biogas production involving several stages such as substrate hydrolysis, acidogenesis, acetogenesis and methanogenesis. In this sense, the next-generation high-throughput sequencing provides a powerful tool for dissecting microbial community structure and methane-producing pathways in anaerobic digestion. Here, a taxonomic and functional metagenomic analysis of microbial community residing in an industrial-scale biogas fermenter has been carried out at different steps of biogas production. Methods Sample were collected from an industrial-scale mesophilic plant, daily fed with maize silage, consisting of a three steps production taking place in a bioreactor, post-reactor and a storage tank. Total DNA was extracted from samples belonging to each stage of biogas production. Metagenomic analysis were carried out by 16S and shotgun sequencing approach. The 16S datasets were generated by sequencing the bacterial and archaeal V4 hypervariable region. Reads from 16S sequencing were aligned against SILVA ribosomal RNA sequence database by using MALT (1), while shotgun reads were aligned against NCBI-nr sequence database by using DIAMOND (2). Taxonomic binning and functional annotation were performed with MEGAN 6 software (3). Results Over 14.5 million high quality reads (about 3.4 gigabases) were generated on the Ion Torrent S5 Sequencing System. About 2.4 and 3 million reads were assigned for 16S and shotgun approach, respectively. Although the average number of assigned taxa for 16S analysis was considerably lower than shotgun analysis, the overall taxa distribution resulting from both sequencing strategies was conserved. In detail, metagenomic analysis revealed that the superkingdom of Bacteria was dominant (~93%) along the production steps, whereas Archaea were less represented (~4%). Within Bacteria the most abundant phyla were Firmicutes, mostly represented by Clostridia, followed by Bacteroidetes, Synergistetes and Proteobacteria. Within the superkingdom of Archaea, only microorganisms belonging to the phylum of Euryarchaeota were detected. Within Euryarchaeota the dominant genera were Methanosarcina and Methanoculleus, notably to be key microorganisms involved in methanogenesis. Data showed that during biogas production steps the abundance of Methanosarcina genus decreased from bioreactor to storage tank, with a simultaneous increase of Methanoculleus genus. Functional analysis of assigned reads also supported a shift from acetotrophic methanogens to hydrogenotrophic methanogens. In this regards, considering the key methanogenesis pathways, in bioreactor most of the assigned reads were related to genes encoding for acetate kinase, acetyl-CoA synthetase and phosphate acetyltransferase, whereas in the storage tank reads were mostly related to genes encoding for formylmethanofuran dehydrogenase, methyle-netetrahydromethanopterin dehydro-genase and methenyltetrahydromethanopterin cyclohydrolase. Conclusions In this work, a metagenomics analysis of an industrial-scale biogas plant has been carried out. The combination of both 16S and shotgun sequencing approach successfully addressed the taxonomical and functional analysis of microbial community, revealing new insights in microbial and functional dynamics during biogas production steps. Acknowledgments This work was supported by H2020-E.U.3.2-678781-MycoKey-Integrated and innovative key actions for mycotoxin management in the food and feed chain and Short Term Mobility Program (2016) of National Research Council. We thanks Austep S.p.A for providing samples. References 1.Herbig A.,Maixner F., Bos K.I., et al., 2016. MALT: fast alignment and analysis of metagenomic DNA sequence data applied to the Tyrolean Iceman. Preprint at http://biorxiv.org/content/early/2016/04/27/050559. 2.Buchfink B., Xie C., Huson D.H., 2015.Fast and sensitive protein alignment using DIAMOND. Nature Methods 12, 59-60. 3.Huson D.H., Beier S., Flade I., et al., 2016. MEGAN Community Edition - Interactive Exploration and Analysis of Large-Scale Microbiome Sequencing Data, PLoS Computational Biology 12(6): e1004957.

Published
2017

9. La dinamicità del sistema simbolico-culturale nella percezione e costruzione della distanza sociale

Author

DI GENNARO, GIACOMO, G. Mulè, CESAREO V., DI GENNARO G, MULE' G, G. Di Gennaro, G. Mulè., V.Cesareo, DI GENNARO, Giacomo, and G., Mulè

Subjects
sistema simbolico culturale, città, integrazione sociale, globalizzazione, Distanza sociale
Abstract

Il contributo si situa in un volume che affronta, mediante una ricerca empirica, la dinamica dei rapporti tra spazio simbolico-culturale e spazio fisico che è alla base dei processi di distanziamento sociale. Specificamente nel saggio sono analizzati i valori, gli orientamenti culturali e le connessioni cognitive che si producono influenzando l'agire sociale. Si discute il presupposto struttural-funzuionalista che interpreta la socializzazione come processo idoneo a integrare funzionalmente sia le agenzie di socializzazione che il sistema delle norme e i valori soggettivamente preferiti. Ciò che si pone in evidenza è, invece, la rottura dell'unitarietà funzionale delle diverse agenzie che operano in direzione dell'orientamento all'integrazione culturale del soggetto, riverberandosi sul distanziamento agito e subito e sul modo in cui vengono percepiti i comportamenti devianti e lo stesso particolare ordine nelle relazioni e nelle interazioni tra gruppi e soggetti. Questo processo si connette fortemente con la centralità dello status e con la fruibilità dello spazio geometrico, ovvero organizzazione fisica dello spazio e significazione cognitiva e simbolica delle interazioni che in esso ricadono.

Published
2007

11. Molecular characterization of Pleurotus eryngii varieties occurring in Italy

Author

A. Villani, E. Galli, C. Paciolla, G. Stea, A. F. Logrieco, C. Siniscalco, G. Mulè, and A. Susca

Subjects
rpb2, ef1-a, Pleurotus eryngii species complex, molecular phylogeny, SNPs
Abstract

The Pleurotus eryngii species complex is an economically important group which includes several closely related varieties, whose genetic discrimination is still not clear. One hundred and ten Italian strains of Pleurotus eryngii belonging to the varieties elaeoselini, eryngii, ferulae and thapsiae and P. nebrodensis were analysed by sequencing two housekeeping genes (ef1-a and rpb2), in order to find molecular markers for the identification of different varieties. Sequence analysis of partial ef1-a and rpb2 genes, allowed identification of some conserved nucleotide positions within each variety but variable among var. elaeoselini, var. eryngii, var. ferulae var. thapsiae and P. nebrodensis, allowing their discrimination. Phylogenetic analysis from the data of the two genes data set showed that var. elaeoselini, var. thapsiae, var. ferulae and var. eryngii are closely related to each other, and confirm P. nebrodensis as a separate clade.

Published
2015
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12. Molecular Diversity and PCR-detection of Toxigenic Fusarium Species and Ochratoxigenic Fungi : Under the Aegis of COST Action 835 ‘Agriculturally Important Toxigenic Fungi 1998–2003’, EU Project (QLK1-CT-1998-01380) and the ISPP ‘Fusarium Committee’

Author

G. Mulè, John A. Bailey, B.M. Cooke, A. Logrieco, G. Mulè, John A. Bailey, B.M. Cooke, and A. Logrieco

Subjects
Fungi, Mycology, Microbiology, Plant diseases, Botany, Microbial ecology, Chemistry, Agriculture
Abstract

Toxigenic Fusarium species and ochratoxigenic fungi are responsible for various plant diseases which have important consequential effects on both human and animal health worldwide. The development of rapid, robust and sensitive detection methods based on new molecular technologies is urgently needed in order to identify fungal contamination in the field and quantify toxin accumulation in food and animal feed. Most of the contributions in this special issue are from results obtained through the EU 5th Framework project (QLKI-CT-1998-01380)'DETOX-FUNGI: early detection of toxigenic Fusarium species and ochratoxi­ genic fungi in plant products', which has strongly stimulated interaction and co-operation between many European scientists. Valuable contributions from other scientists have guaranteed a complete overview of this stimulating and interesting topic. This is the third special issue published in the European Journal of Plant Pathology concerning my­ cotoxigenic fungi under the aegis of COST Action 835'Agriculturally Important Toxigenic Fungi'. The first two dealt with'Mycotoxins in Plant Disease'(Vol. 108(7) 2002) and'Epidemiology of Mycotoxin Pro­ ducing Fungi'(Vol. 109(7) 2003). The present issue contains contributions which cover aspects of molecular diversity, phylogeny and PCR-detection of toxigenic Fusarium species and various ochratoxigenic fungi. We hope these will prove helpful to researchers involved in similar work and will stimulate the future studies required for the early detection of these fungi, which is so essential for overcoming the health risks associated with mycotoxin-contaminated food products.

Published
2012

13. Marcatori molecolari per la tracciabilità del cardoncello

Author

A. Villani, G. Mulè, A. Susca, L. Mastropasqua, and C. Paciolla

Subjects
molecular markers Pleurotus eryngii species complex, enzyme analysis
Abstract

In this work the authors analyzed sixty italian strains of the edible mushroom "cardoncello" (Pleurotus eryngii) belonging to the varieties eryngii and ferulae by sequencing two housekeeping genes (ef1-a and rpb2) and evaluating the activity of peroxidase, superoxide dismutase and catalase, in order to find some molecular markers for the traceability of "cardoncello". Sequence analysis showed the presence of Snps variety-specific (eryngii and ferulae) in both genes useful for the development of molecular identification tools. none correlation between enzymatic analysis and analyzed varieties was observed.

Published
2014

19. MOLECULAR AND CHEMICAL CHARACTERIZATION OF THE PATHOSYSTEM FUSARIUM VERTICILLIOIDESMAIZE

Author

G. Mulè, I. Lazzaro, A. Susca, A. Ritieni, A. Lanubile, J. Bernardi, A. Marocco, and P. Battilani

Abstract

A two-year study on the pathosystem F. verticillioides-maize was conducted speculating on both in vitro and in planta perspectives. The former studied the effects of temperature (T) and water activity (aw) on fumonisin B (FBs) production and expression of FUM2 and FUM21 genes in ITEM 10027 and 1744 F. verticillioides strains grown up to 21 days. The latter monitored which genes were differentially expressed in resistant and susceptible maize lines after 2-3 days infection by ITEM 1744. The in vitro study showed that ITEM 10027 was the highest FBs producer, with predominance of FB1. The maximum level of FB was registered after 21 days in both strains. FUM2 and FUM21 were expressed in all studied conditions, with a 10X difference between the former and the latter. The peak of transcription level was reached after 14 days of incubation for both FUM genes. No data were collected on cultures grown at fixed aw=0.900 because the fungus did not grow at all temperatures tested till 21 days of incubation. The in planta study showed that resistant lines tested were poorly infected by ITEM 1744. Genes differentially expressed were divided into 11 functional categories and nearly 10% was assigned to the class "cell rescue, defence and virulence". Most of the pathogenesis-related genes were differentially activated after fungal infection in relation to the resistance level of maize genotypes. In the resistant kernels, defence-related genes provided basal protection against the fungus, while in the susceptible kernels, the same genes were induced specifically after pathogen attack.

Published
2011

20. Within-field variability of Fusarium ear blight and its associated mycotoxins

Author

Xu X.M., Parry D.W., Nicholson P., Thomsett M.A., Simpson D., Edwards S.G., Cooke B.M., Doohan F.M., Monaghan S., Moretti A., Tocco G., Mulè G., Hornok L., Béki E., Tatnell J., and Ritieni A.

Subjects
Wheat head scab, NIV, food and beverages, Fungal DNA - Aggregation, DON, Chaff - Flour
Abstract

Within-field variability in the Fusarium head blight (FHB) and its associated mycotoxins was studied in four European countries. At each of 14 sites, each FHB pathogen and associated mycotoxins were quantified in 16 quadrat samples at harvest. Overall, the incidence of quadrat samples with detectable and quantifiable pathogen DNA was significantly lower in the grain than in the corresponding chaff. Deoxynivalenol (DON) was the most frequently detected toxin in the samples and its accumulation was most strongly associated with the presence of Fusarium graminearum. Nivalenol (NIV) accumulation was significantly associated only with the presence of F. culmorum. Zearalenone (ZON) accumulation was strongly associated with the presence of all three pathogens (F. graminearum, F. culmorum and F. poae). The levels of both DON and ZON concentrations were positively related to the amount of F. graminearum DNA in the grain or in the chaff. The presence/absence of FHB pathogens within a single quadrat appeared to be independent of each other. The presence of a particular FHB pathogen and the amount of its DNA, as well as the associated mycotoxin(s), varied greatly among samples at each site. This study demonstrated the large extent of within-field variability of FHB and its associated mycotoxins, and the importance of representative sampling in FHB studies.

Published
2008
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25. Mitochondrial DNA diversity and lineage determination of European isolates of Fusarium graminearum (Gibberella zeae)

Author

M. Láday, Á. Juhász, G. Mulè, A. Moretti, Á. Szécsi, and A. Logrieco

Subjects
identification, Plant Science, Horticulture, lineage distribution, mtDNA RFLP, Agronomy and Crop Science, scab
Abstract

Restriction fragment length polymorphisms (RFLPs) were used to assess genetic diversity of mitochondrial DNA (mtDNA) among standard isolates of seven lineages of Fusarium graminearum. The mtDNA patterns within each lineage were very similar (>89%), whereas significant differences were observed between the isolates belonging to different lineages, with the exception of lineages 1 and 4 where strong similarity was found between the RFLPs. Analysis of different band patterns resulted in characteristic HhaI and HaeIII bands that were suitable for identification of members of lineages 7, 6, 5, 3 and 2. Investigation of lineage distribution of 144 European isolates revealed that 142 belong to lineage 7. These data, therefore, confirmed the hypothesis that members of lineage 7 are predominant in Europe. Further analysis of isolates belonging to lineage 7 resulted in five haplotypes. These haplotypes have arisen as different combinations of three RFLP patterns for both HaeIII and HhaI restriction enzymes. Two isolates from Hungary, however, shared the same mtDNA RFLP profiles with a standard isolate of lineage 3, indicating that members of lineage 3, at a lower frequency, may also occur in Europe.

Published
2004
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32. [Arterial hypertension caused by low renin: comparison of the calcium antagonist nifedipine and the ACE-inhibitor enalapril as to antihypertensive efficacy]

Author

G, Andronico, M B, Carone, G, Mulè, M T, Mangano, and G, Cerasola

Subjects
Male, Diabetes Mellitus, Type 2, Enalapril, Nifedipine, Hypertension, Renin, Sodium, Humans, Blood Pressure, Female, Middle Aged
Abstract

The aim of this study was to compare the clinical effects of calcium-entry blocker Nifedipine and ACE-inhibitor Enalapril in hypertensive patients with glucose intolerance that have lower plasma renin activity. A blood sample for basal PRA was obtained from 21 subjects; then, 11 patients received Nifedipine (20 mg. b.i.d.) and 10 Enalapril (20 mg. q.d.). The extent of blood pressure fall after 12 weeks of treatment was inversely related to basal PRA levels in Nifedipine treated group only; however, the hypotensive effect of both drugs was comparable.

Published
1990

33. Mitochondrial DNA Variability in Fusarium Proliferatum (Gibberella Intermedia).

Author

M. Láday, G. Mulè, A. Moretti, Z. Hamari, Á. Juhász, Á. Szécsi, and A. Logrieco

Abstract

Restriction fragment length polymorphisms (RFLP) were used to assess genetic diversity of mitochondrial DNA (mtDNA) among 184 isolates of Fusarium proliferatum recovered from maize, asparagus, palms and reed. All strains were cross-fertile with standard mating type tester strains of Gibberella intermedia. Sixteen mitochondrial haplotypes were identified following digestion of DNAs with HaeIII, with seven, seven, five and six different haplotypes from maize, asparagus, palms and reed, respectively. Four haplotypes (I, III, IV and VII) were found on more than one host. Of these four, haplotype I was dominant on maize, representing 71% of the isolates. The banding patterns for haplotypes III and IV were >90% similar to the banding pattern of haplotype I. Haplotypes I, III and IV accounted for 87% of the isolates from maize, but were less common on the other hosts, accounting for 70%, 52% and 33% of the isolates from asparagus, palms and reed, respectively. Thirteen of the 16 haplotypes were recovered from only a single host plant species. When comparing the banding patterns and frequencies of these haplotypes, at least five were recovered at a higher frequency from one host relative to the others. Our results suggest that mtDNA RFLP analysis is a useful indicator of genetic divergence in Fusarium proliferatum. [ABSTRACT FROM AUTHOR]

Published
2004

34. A Species-Specific PCR Assay Based on the Calmodulin Partial Gene for Identification of Fusarium Verticillioides, F. Proliferatum and F. Subglutinans.

Author

G. Mulè, A. Susca, G. Stea, and A. Moretti

Abstract

Fusarium proliferatum, F. subglutinans and F. verticillioides are the most important Fusarium species occurring on maize world-wide, capable of producing a wide range of mycotoxins which are a potential health hazard for animals and humans. The ribosomal internal transcribed spacer and a portion of the calmodulin gene were sequenced and analysed in order to design species-specific primers useful for diagnosis. The primer pairs were based on a partial calmodulin gene sequence. Three pairs of primers (PRO1/2, SUB1/2 and VER 1/2) produced PCR products of 585, 631 and 578bp for F. proliferatum, F. subglutinans and F. verticillioides, respectively. Primer specificity was confirmed by analyzing DNA of 150 strains of these species, mostly isolated from maize in Europe and USA. The sensitivity of primers was 12.5 pg when the pure total genomic DNA of each species was analyzed. The developed PCR assay should provide a powerful tool for the detection of toxigenic fungi in maize kernels. [ABSTRACT FROM AUTHOR]

Published
2004

35. Toxin Profile, Fertility and AFLP Analysis of Fusarium verticillioides from Banana Fruits.

Author

A. Moretti, G. Mulè, A. Susca, M.T. González-Jaén, and A. Logrieco

Abstract

Gibberella fujikuroi is composed of at least nine mating populations (MPs), corresponding to biological species and assigned letters (from A to I). Each MP possesses a specific toxicological profile and a preferential host. Members of Fusarium verticillioides and F. thapsinum, anamorphs respectively of MPs A (G. moniliformis) and F (G. thapsina), share identical morphological traits, but they have a different preferential hosts (maize and sorghum, respectively) and toxin profiles, beingable the only member of MP A to produce fumonisins and the only member of MP F to produce moniliformin. Isolates from banana fruits were identified morphologically as F. verticillioides. The isolates were analyzed for fumonisin and moniliformin production. While none of the isolates produced fumonisin, all the isolates produced moniliformin. The isolates were crossed with tester strains of MPs A and F, showing ability to produce fertile perithecia only when crossed by MP A tester strains isolated from maize. However, the time required for the formation of fertile perithecia and their size differed significantly from the usual fertile crosses of strains belonging to MP A. Pathogenicity tests using such isolates of F. verticillioides isolated from banana and a set of F. verticillioides isolates isolated from maize were also performed on banana fruits. The data showed that the isolates from banana were more pathogenic. Finally, isolates from banana and maize were compared using AFLP. The results revealed that isolates from banana and maize produced two distinctly different clusters. In conclusion, isolates of F. verticillioides from banana showed specific traits (toxin production, in vitro fertility, pathogenicity and molecular profiles), that were different to those of the same species from maize. This could reflect important differences in the ecology and natural history of the population from banana and should encourage further investigations into the mechanisms of toxin production and pathogenicity within the same MP. [ABSTRACT FROM AUTHOR]

Published
2004

36. Identification of Growth Stage Specific Transcript Profiles in Fusarium Proliferatum (Gibberella Fujikuroi, Mating Population D) by cDNA-AFLP Analysis.

Author

A. Jeney, E. Béki, G. Mulè, and L. Hornok

Abstract

A cDNA-AFLP approach was developed to identify differentially regulated genes in mycotoxin-producing and non-producing growth stages of Fusarium proliferatum(Gibberella fujikuori Mating Population-D). Using 160 PCR-primer combinations, a number of fragments which showed strikingly different intensities depending on growth stage were cloned, subjected to Northern analysis and sequenced. Two fragments, amplified from genes up-regulated during spore germination gave significant sequence homology to an amino acid transporter gene from Neurospora crassa and a GAL4-like transcriptional activator, respectively. Among the cDNAs derived from late growth stage transcripts, fragments of genes involved in polyol metabolism and cell cycle regulation, respectively were identified. Additional cDNAs (without significant homologies to known sequences) were also isolated and these clearly differentiated between mycotoxin-producing and non-producing growth stages of the fungus. [ABSTRACT FROM AUTHOR]

Published
2004

37. Mitochondrial DNA Diversity and Lineage Determination of European Isolates of Fusarium graminearum (Gibberella zeae).

Author

M. Láday, Á. Juhász, G. Mulè, A. Moretti, Á. Szécsi, and A. Logrieco

Abstract

Restriction fragment length polymorphisms (RFLPs) were used to assess genetic diversity of mitochondrial DNA (mtDNA) among standard isolates of seven lineages of Fusarium graminearum. The mtDNA patterns within each lineage were very similar (>89%), whereas significant differences were observed between the isolates belonging to different lineages, with the exception of lineages 1 and 4 where strong similarity was found between the RFLPs. Analysis of different band patterns resulted in characteristic HhaI and HaeIII bands that were suitable for identification of members of lineages 7, 6, 5, 3 and 2. Investigation of lineage distribution of 144 European isolates revealed that 142 belong to lineage 7. These data, therefore, confirmed the hypothesis that members of lineage 7 are predominant in Europe. Further analysis of isolates belonging to lineage 7 resulted in five haplotypes. These haplotypes have arisen as different combinations of three RFLP patterns for both HaeIII and HhaI restriction enzymes. Two isolates from Hungary, however, shared the same mtDNA RFLP profiles with a standard isolate of lineage 3, indicating that members of lineage 3, at a lower frequency, may also occur in Europe. [ABSTRACT FROM AUTHOR]

Published
2004

38. PCR Assay for Identification of Aspergillus Carbonarius and Aspergillus Japonicus.

Author

G. Perrone, A. Susca, G. Stea, and G. Mulè

Abstract

Black Aspergilli, and in particular Aspergillus carbonarius, are the main causes of contamination of grapes and their by-products by ochratoxin A. A PCR-based method was developed to detect DNA of A. carbonarius and A. japonicus. Two pairs of primers (CARBO1/2 and JAPO1/2) designed from the calmodulin gene, produced PCR products of 371 and 583 bp for A. carbonarius and A. japonicus, respectively. Primer specificity was tested with DNA of 107 strains belonging to Aspergillus section Nigri isolated mostly from grapes in Europe. The sensitivity of primers CARBO1/2 and JAPO1/2 was 12.5 pg when using pure total genomic DNA of the two species. The developed primers provide a powerful tool for detection of the main ochratoxigenic producing Aspergillus species in grapes. [ABSTRACT FROM AUTHOR]

Published
2004

39. Monitoring fungi and mycotoxin potential in pistachio nuts of Turkish origin: A snap-shot for climate change scenario.

Author

Anelli P, Haidukowski M, Ferrara M, Kisikkaya A, Pembeci C, Ozer H, Mulè G, Loi M, Moretti A, and Susca A

Subjects
Turkey, Aflatoxins metabolism, Aflatoxins analysis, Nuts microbiology, Aspergillus flavus genetics, Aspergillus flavus metabolism, Aspergillus flavus classification, Aspergillus flavus isolation & purification, Pistacia microbiology, Mycotoxins metabolism, Mycotoxins analysis, Climate Change, Aspergillus metabolism, Aspergillus genetics, Aspergillus isolation & purification, Aspergillus classification
Abstract

Pistachio (Pistacia vera L.) is an economically important tree nut. Due to its nutritional properties and health benefits, it is considered a healthy food and thus widely consumed worldwide. However, fungal contamination of the commodities has received considerable attention because of possible contamination by toxigenic fungi, important source of mycotoxins, resulting from secondary metabolism and hazards to health consumer. Members of the genus Aspergillus, mainly Aspergillus flavus and Aspergillus niger, are reported as occurring most frequently on pistachio nuts, because able to grow in the presence of low amounts of water and to produce mycotoxins (aflatoxins and ochratoxins), that are well known for their harmful health effects on humans. Monitoring the contaminating fungal species is particularly worthy of note also in climate change scenario, allowing to notice changes in fungal population composition through the time. This study aimed to contribute to collect data about fungal population and mycotoxins occurred in pistachio samples collected in Turkey: prevalence of 2 species, A. flavus and Aspergillus tubingensis, was assessed. The A. flavus strains consisted of a mixed population of aflatoxin producers and non-producing strains in vitro, with evidence of a new genotype in gene cluster within strains of aflatoxin non-producing chemotype., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Published
2024
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40. Relationship Between Short-Term Blood Pressure Variability and Choroidal-Retinal Thicknesses Assessed by Optical Coherence Tomography in Hypertensive Subjects.

Author

Carollo C, Vadalà M, Ferrara M, Chisci E, La Felice A, Di Natale KV, Sorce A, Bonfiglio VME, and Mulè G

Abstract

Background/Objectives: The complications of hypertension depend not only on the mean blood pressure (BP) but also on its variability (BPV). Recent studies suggest that the choroid may serve as an indicator of systemic vascular damage. These studies have been made possible by the increased availability of optical coherence tomography (OCT). The aim of our study was to analyze the relationship between short-term BP variability (STBPV) and choroid-retinal thickness in hypertensive patients. Methods: A total of 98 patients with a mean age of 49 ± 12 years were enrolled in the study. All participants underwent 24 h blood pressure (BP) monitoring to measure 24 h mean systolic (SBP) and diastolic blood pressure (DBP), along with their respective standard deviations (SD), the weighted SD of 24 h SBP and DBP, and the average real variability (ARV) of 24 h SBP and DBP. The choroid-retinal region was assessed using Swept-Source OCT, with choroidal thickness (ChT) and retinal thickness divided into three concentric rings, and their mean choroidal thickness (ChT-or) was calculated. Results: The choroidal thickness of the concentric rings was found to be inversely correlated with all ARV values of the monitored blood pressure means. In particular, a correlation was observed between the ARV of daytime DBP and ChT-or. This correlation remained statistically significant (β = -0.34; p = 0.02) even after adjustment for various confounding factors. The ARV of daytime DBP was the only STBPV index to maintain a significant association, in the multivariate analysis, with the central ring mean thickness (β = -0.314; p = 0.001) and the inner choroidal ring mean thickness (β = -0.262; p = 0.003). Conclusions: Our study demonstrated an independent negative association between short-term BP variability (STBPV), when expressed as ARV of daytime DBP, and choroidal thickness. This finding confirms the value of choroidal thickness as a marker of cardiovascular risk.

Published
2024
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41. Donor-derived mold infections in lung transplant recipients: The importance of active surveillance.

Author

Mularoni A, Cona A, Coniglione G, Barbera F, Di Martino G, Mulè G, Campanella M, Di Mento G, Nunnari G, Grossi PA, Sanguinetti M, Mikulska M, De Carolis E, and Bertani A

Subjects
Humans, Male, Middle Aged, Prospective Studies, Female, Adult, Antifungal Agents therapeutic use, Incidence, Mycoses epidemiology, Mycoses microbiology, Aged, Rhizopus isolation & purification, Aspergillus niger isolation & purification, Fungi isolation & purification, Lung Transplantation adverse effects, Tissue Donors, Transplant Recipients statistics & numerical data
Abstract

Unexpected donor-derived fungal infections represent a rare but potentially fatal complication in lung transplant (Tx) recipients. Timely communication of the results of donor cultures and prompt treatment of recipients are crucial to mitigate the consequences of donor-derived transmissions. In this prospective cohort study, all consecutive patients who underwent lung transplantation from 2015 to 2022 were included. In December 2015, a Local Active Surveillance System has been implemented to provide biovigilance of donor culture results and optimize recipients' management. The aim of this study is to investigate the incidence of unexpected, mold-positive cultures among lung donors and the rate of transmission to recipients. Furthermore, management strategies and outcome of recipients with mold transmission are described. In case of isolation of the same mold in donor and recipient cultures, when possible, transmission was confirmed by dendrogram analysis. During the study period, 82 lung Tx were performed from 80 donors. The prevalence of donors with "unexpected" mold isolation from the respiratory tract was 3.75% (3/80). Isolated molds were Aspergillus niger, Rhizopus oryzae, and Aspergillus flavus. Transmissions occurred in all the three cases (100%) with a mean time of 5 days from lung Tx but none of the recipients developed invasive mold disease. Our Local Active Surveillance System allowed prompt recognition of lung donors unexpected mold colonization. Even though transmission occurred, introduction of early targeted antifungal therapy prevented potential catastrophic consequence of mold donor-derived infection in the immediate post-Tx period., (© 2024 The Author(s). Transplant Infectious Disease published by Wiley Periodicals LLC.)

Published
2024
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42. Light emitting diodes for the improvement of postharvest quality of wild rocket in soilless and soil-bound cultivation systems.

Author

Loi M, De Leonardis S, Mulè G, Serio F, Bottiglione B, Paciolla C, and Villani A

Abstract

Wild rocket ( Diplotaxis tenuifolia (L.) DC cv. Dallas) is a leafy green vegetable appreciated for its pungent taste and healthy properties, often consumed as a ready-to-eat product. The cultivation system is crucial in determining the overall quality, while postharvest storage is fundamental for preserving nutritional quality, phytochemicals, and vitamins. This study aimed to investigate the phytochemical content and microbiological quality of soilless (SS) and soil-bound (SB) wild rocket during cold postharvest storage under blue, red, and green Light Emitting Diode (LED). Blue LED increased chlorophylls and carotenoids in SB after two days of storage, and chlorophyll a in SS after seven days. Furthermore, it reduced H 2 O 2 levels after two days (SS and SB) and lipid peroxidation in SB. Red LED increased phenols in both SS and SB but was detrimental to chlorophyll, carotenoids, and oxidative markers. Green LED had less significant effects. Microbiological growth varied with LED treatment: green light increased mesophilic bacteria in SB, and red light did so in SS by day four, while blue light reduced bacterial growth at the end of storage. Overall, Blue LED was the most effective LED in preserving postharvest quality. Soilless cultivation was particularly beneficial in reducing lipid peroxidation and maintaining cell membrane integrity during long-term storage, and it might also be more effective in preserving ascorbic acid. Conversely, soil-bound cultivation methods could enhance initial polyphenol content or better preserve it during early storage. This study highlights the complex interplay of pre-harvest conditions, postharvest quality, and shelf-life performance., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Author(s).)

Published
2024
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43. Performance evaluation of a self-administered point-of-care test for anal HPV screening in PrEP users: data from a community-based PrEP service.

Author

Biasioli L, Rossotti R, Tavelli A, De Bona A, Tincati C, Calzavara D, Vinti P, Baiguera C, D'Amico F, Nava A, Repossi R, Bossolasco S, Muccini C, Mulè G, Tesoro D, d'Arminio Monforte A, and Cernuschi M

Subjects
Humans, Male, Adult, Female, Mass Screening methods, Papillomaviridae genetics, Papillomaviridae isolation & purification, Anal Canal virology, Feasibility Studies, Middle Aged, Homosexuality, Male statistics & numerical data, HIV Infections diagnosis, HIV Infections prevention & control, Young Adult, Self-Testing, Papillomavirus Infections diagnosis, Papillomavirus Infections prevention & control, Point-of-Care Testing, Pre-Exposure Prophylaxis, Sensitivity and Specificity
Abstract

Objectives: In this study, we compared the performance of a self-administered point-of-care test (POCT) for anal human papillomavirus (HPV) screening with laboratory gold-standard test in pre-exposure prophylaxis (PrEP) users and evaluated its feasibility., Methods: We enrolled PrEP users from a local community-based PrEP service. Each participant self-collected an anal swab to test anal HPV with a PCR POCT capable of detecting 14 high-risk HPV genotypes. Anonymous questionnaires on self-sampling feasibility were completed. Participants were then referred to local clinics to undergo standard viral genotyping. Concordance between POCT and gold-standard test was measured with absolute agreement and Cohen's kappa. Receiver operating characteristic (ROC) curves were used to calculate POCT sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV)., Results: 179 subjects got a valid POCT result, most of them men (98.3%) and men who have sex with men (90.4%). 68.2% tested positive for at least one high-risk HPV genotype on POCT. 150 feasibility questionnaires were collected: 92.7% of compilers found the self-swab easy to perform. For 178 subjects, a gold-standard test valid result was also available: 77% tested positive for at least one high-risk HPV genotype. The median time elapsed between the two tests was 9.8 months, due to COVID-19-related service interruptions. Agreement between POCT and gold-standard test was 79.3% (Cohen's kappa=0.49). POCT showed a sensitivity of 81.0%, a specificity of 73.8%, a PPV of 91.0% and an NPV of 54.4%., Conclusions: POCT showed a moderate agreement with gold-standard test and a discrete sensitivity and specificity, suggesting that it could be a useful and feasible additional tool for HPV screening, especially in low-resource and community-based settings., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. No commercial re-use. See rights and permissions. Published by BMJ.)

Published
2024
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44. May Measure Month 2022 in Italy: A Focus on Fixed-dose Combination, Therapeutic Adherence, and Medical Inertia in a Nationwide Survey.

Author

Del Pinto R, Agabiti Rosei C, Borghi C, Cipollini F, Cottone S, De Giorgi GA, Di Guardo A, Dugnani M, Fabris B, Giannattasio C, Giacchetti G, Minuz P, Mulè G, Nazzaro P, Parati G, Rattazzi M, Saladini F, Salvetti M, Sarzani R, Savoia C, Tocci G, Veglio F, Volpe M, Vulpis V, Baldini G, Ferri C, and Muiesan ML

Subjects
Humans, Female, Male, Italy epidemiology, Middle Aged, Cross-Sectional Studies, Aged, Treatment Outcome, Practice Patterns, Physicians', Time Factors, Adult, Attitude of Health Personnel, Antihypertensive Agents therapeutic use, Antihypertensive Agents adverse effects, Antihypertensive Agents administration & dosage, Hypertension drug therapy, Hypertension physiopathology, Hypertension epidemiology, Hypertension diagnosis, Medication Adherence, Blood Pressure drug effects, Health Knowledge, Attitudes, Practice, Health Care Surveys, Drug Combinations
Abstract

Introduction: Hypertension is the main risk factor for cardiovascular diseases (CVD). Notably, only about half of hypertensive patients manage to achieve the recommended blood pressure (BP) control. Main reasons for the persistence of uncontrolled BP during treatment are lack of compliance on the patients' side, and therapeutic inertia on physicians' side., Methods: During the global BP screening campaign "May Measure Month" (MMM) (May 1st to July 31st, 2022), a nationwide, cross-sectional, opportunistic study endorsed by the Italian Society of Hypertension was conducted on volunteer adults ≥ 18 years to raise awareness of the health issues surrounding high BP. A questionnaire on demographic/clinical features and questions on the use of fixed-dose single-pills for the treatment of hypertension was administered. BP was measured with standard procedures., Results: A total of 1612 participants (mean age 60.0±15.41 years; 44.7% women) were enrolled. Their mean BP was 128.5±18.1/77.1±10.4 mmHg. About half of participants were sedentary, or overweight/obese, or hypertensive. 55.5% individuals with complete BP assessment had uncontrolled hypertension. Most were not on a fixed-dose combination of antihypertensive drugs and did not regularly measure BP at home. Self-reported adherence to BP medications was similar between individuals with controlled and uncontrolled BP (95% vs 95.5%)., Conclusions: This survey identified a remarkable degree of therapeutic inertia and poor patients' involvement in the therapeutic process and its monitoring in the examined population, underlining the importance of prevention campaigns to identify areas of unsatisfactory management of hypertension, to increase risk factors' awareness in the population with the final purpose of reducing cardiovascular risk., (© 2024. The Author(s).)

Published
2024
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45. DypB peroxidase for aflatoxin removal: New insights into the toxin degradation process.

Author

Mangini V, Rosini E, Caliandro R, Mangiatordi GF, Delre P, Sciancalepore AG, Pollegioni L, Haidukowski M, Mazzorana M, Sumarah MW, Renaud JB, Flaig R, Mulè G, Belviso BD, and Loi M

Subjects
Peroxidases metabolism, Aflatoxin B1 metabolism, Coloring Agents chemistry, Peroxidase, Aflatoxins
Abstract

Aflatoxin B 1 (AFB 1 ) is one of the most potent carcinogens and a widespread food and feed contaminant. As for other toxins, many efforts are devoted to find efficient and environmentally-friendly methods to degrade AFB 1 , such as enzymatic treatments, thus improving the safety of food and feed products. In this regard, the dye decolorizing peroxidase of type B (DypB) can efficiently degrade AFB 1 . The molecular mechanism, which is required to drive protein optimization in view of the usage of DypB as a mycotoxin reduction agent in large scale application, is unknown. Here, we focused on the role of four DypB residues in the degradation of AFB 1 by alanine-scanning (residues 156, 215, 239 and 246), which were identified from biochemical assays to be kinetically relevant for the degradation. As a result of DypB degradation, AFB 1 is converted into four products. Interestingly, the relative abundancy of these products depends on the replaced residues. Molecular dynamics simulations were used to investigate the role of these residues in the binding step between protein and manganese, a metal ion which is expected to be involved in the degradation process. We found that the size of the haem pocket as well as conformational changes in the protein structure could play a role in determining the kinetics of AFB 1 removal and, consequently, guide the process towards specific degradation products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Published
2024
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46. Evaluation of the prevalence of dental agenesis through the use of orthopantomography in a sample of subjects residing in Lombardy and Piedmont regions.

Author

Nobili A, Butti AC, Mulè G, Clivio A, and Re D

Subjects
Humans, Child, Adolescent, Prevalence, Radiography, Panoramic, Italy, Molar, Third
Abstract

Aim: Dental agenesis is one of the most frequent dental anomalies, with a prevalence varying from 1.6% to 36.5%, depending on the populations studied. The patient's age at diagnosis, sex, and ethnic differences are considered possible influenting factors that can explain such a wide range of prevalence. The objective of the study was to define the frequency of dental agenesis in a sample of subjects living in Piedmont and Lombardy regions of Italy., Materials: X-rays, already taken for other diagnostic purposes, were collected. Orthopantomographies belonging to subjects born after 1995 and aged between 7.9 and 16.9 years were selected. It was assessed the presence of each tooth, except for third molars since they are frequently absent due to their variability. If a tooth was missing and the patient had additional radiographs, the other radiographs were evaluated to confirm the diagnosis or to rule out a delayed calcification or the presence of a malposition tooth., Results: Orthopantomographies were collected from 1,020 subjects and 98 of them presented agenesis, with a prevalence of 5% for females and 4.61% for males. The most affected teeth were 35 and 45, followed by 12 and 22. The lower arch was more frequently involved by agenesis: there were 107 teeth absent in the mandibular arch and 83 in the maxillary arch.

Published
2023
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47. Aflatoxin B 1 Degradation by Ery4 Laccase: From In Vitro to Contaminated Corn.

Author

Loi M, De Leonardis S, Ciasca B, Paciolla C, Mulè G, and Haidukowski M

Subjects
Zea mays metabolism, Hydrogen Peroxide, Laccase, Aflatoxin B1 metabolism, Aflatoxins metabolism
Abstract

Aflatoxins (AFs) are toxic secondary metabolites produced by Aspergillus spp. and are found in food and feed as contaminants worldwide. Due to climate change, AFs occurrence is expected to increase also in western Europe. Therefore, to ensure food and feed safety, it is mandatory to develop green technologies for AFs reduction in contaminated matrices. With this regard, enzymatic degradation is an effective and environmentally friendly approach under mild operational conditions and with minor impact on the food and feed matrix. In this work, Ery4 laccase, acetosyringone, ascorbic acid, and dehydroascorbic acid were investigated in vitro, then applied in artificially contaminated corn for AFB 1 reduction. AFB 1 (0.1 µg/mL) was completely removed in vitro and reduced by 26% in corn. Several degradation products were detected in vitro by UHPLC-HRMS and likely corresponded to AFQ 1 , epi-AFQ 1 , AFB 1 -diol, or AFB 1 dialehyde, AFB 2a , and AFM 1 . Protein content was not altered by the enzymatic treatment, while slightly higher levels of lipid peroxidation and H 2 O 2 were detected. Although further studies are needed to improve AFB 1 reduction and reduce the impact of this treatment in corn, the results of this study are promising and suggest that Ery4 laccase can be effectively applied for the reduction in AFB 1 in corn.

Published
2023
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48. Characterization of Dextran Produced by the Food-Related Strain Weissella cibaria C43-11 and of the Relevant Dextransucrase Gene.

Author

De Bellis P, Ferrara M, Bavaro AR, Linsalata V, Di Biase M, Musio B, Gallo V, Mulè G, and Valerio F

Abstract

A metabolic feature of lactic acid bacteria (LAB) is the production of exopolysaccharides (EPSs), which have technological and functional properties of interest to the food sector. The present study focused on the characterization of the Weissella cibaria strain C43-11, a high EPS producer in the presence of sucrose, in comparison with a low-producing strain (C2-32), and on possible genetic regulatory elements responsible for the modulation of dextransucrase ( dsr ) genes expression. NMR analysis of the polymeric material produced by the C43-11 strain indicated the presence of dextran consisting mainly of a linear scaffold formed by α-(1-6) glycosidic linkages and a smaller amounts of branches derived from α-(1-2), α-(1-3), and α-(1-4) linkages. Molecular analysis of the dsr genes and the putative transcriptional promoters of the two strains showed differences in their regulatory regions. Such variations may have a role in the modulation of dsr expression levels in the presence of sucrose. The strong upregulation of the dsr gene in the C43-11 strain resulted in a high accumulation of EPS. This is the first report showing differences in the regulatory elements of the dsr gene in W. cibaria and indicates a new perspective of investigation to identify the regulatory mechanism of EPS production.

Published
2022
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49. Effects of an experimental short-time high-intensity warm-up on explosive muscle strength performance in soccer players: A pilot study.

Author

Patti A, Giustino V, Hirose N, Messina G, Cataldi S, Grigoli G, Marchese A, Mulè G, Drid P, Palma A, and Bianco A

Abstract

Objective: This study aimed to evaluate the effects of an experimental short-time warm-up consisting of a small number of intermittent high-intensity sprints on explosive muscle strength performance in soccer players and to identify recovery times after performing the sprints. Furthermore, we evaluated the reliability of a smartphone app in jumping performance. Methods: Twenty male soccer players were given the following tests: 1) the counter-movement jump (CMJ) test with the Microgate system, 2) the counter-movement jump (CMJ) test with the MyJump smartphone app, and 3) the handgrip strength test. The experimental short-time high-intensity warm-up was carried out 1 week after test administration. The warm-up consisted of three maximum sprints over 60 m with 120 s of recovery between sprints. Then, the tests were administered again: the vertical jump height (VJH) performances (five trials) were measured 90 s after the last sprint; the handgrip strength performances (three trials) were measured 120 s after the last vertical jump test. Results: The maximum VJH was found in the third trial of the CMJ test, 330 s after the last sprint ( p < 0.01), the result closest to the baseline. The lowest VJH was found in the first trial of the CMJ test, 90 s after the last sprint ( p < 0.05). Pearson's analysis between the CMJ test with the Microgate system and the CMJ test with MyJump showed a strong correlation (R = 0.96). Lin's concordance correlation coefficient showed a substantial concordance (ρc = 0.959) between measures. Conclusion: This experimental short-time warm-up of high-intensity intermittent sprints appears to be a simple, quick, and efficient activity to accelerate soccer players' optimal performance., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Patti, Giustino, Hirose, Messina, Cataldi, Grigoli, Marchese, Mulè, Drid, Palma and Bianco.)

Published
2022
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50. Identification and Characterisation of pST1023 A Mosaic, Multidrug-Resistant and Mobilisable IncR Plasmid.

Author

Calia C, Oliva M, Ferrara M, Minervini CF, Scrascia M, Monno R, Mulè G, Cumbo C, Marzella A, and Pazzani C

Abstract

We report the identification and characterisation of a mosaic, multidrug-resistant and mobilisable IncR plasmid (pST1023) detected in Salmonella ST1023, a monophasic variant 4,[5],12:i: strain of widespread pandemic lineage, reported as a Southern European clone. pST1023 contains exogenous DNA regions, principally gained from pSLT-derivatives and IncI1 plasmids. Acquisition from IncI1 included oriT and nikAB and these conferred the ability to be mobilisable in the presence of a helper plasmid, as we demonstrated with the conjugative plasmids pST1007-1D (IncFII) or pVC1035 (IncC). A sul3 -associated class 1 integron, conferring resistance to aminoglycosides, chloramphenicol and trimethoprim-sulphonamides, was also embedded in the acquired IncI1 DNA segment. pST1023 also harboured an additional site-specific recombination system ( rfsF / rsdB ) and IS elements of the IS 1 , IS 5 (IS 903 group) and IS 6 families. Four of the six IS 26 elements present constituted two pseudo-compound-transposons, named PCT- sil and PCT-Tn 10 (identified here for the first time). The study further highlighted the mosaic genetic architecture and the clinical importance of IncR plasmids. Moreover, it provides the first experimental data on the ability of IncR plasmids to be mobilised and their potential role in the horizontal spread of antimicrobial-resistant genes.

Published
2022
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