Ανάπτυξη βιοαισθητήρων για τον ταυτόχρονο προσδιορισμό μυκοτοξινών

Οι μυκοτοξίνες είναι χημικές ενώσεις που παράγονται από διάφορες οικογένειες μυκήτων των οποίων η συγκέντρωση επιβάλλεται να προσδιορίζεται σε αρκετές κατηγορίες τροφίμων, λόγω των αρνητικών επιπτώσεων που έχουν στην υγεία των ανθρώπων και των ζώων. Οι μυκοτοξίνες που συνήθως ανιχνεύονται στα τρόφιμα ανήκουν κυρίως σε τρεις κατηγορίες: αφλατοξίνες, φουμονισίνες, και τριχοθηκίνες, με κύριους εκπροσώπους αυτών την αφλατοξίνη Β1 (AFB1), τη φουμονισίνη Β1 (FB1) και τη δεοξυνιβαλενόλη (DON). Στο πλαίσιο αυτό, σκοπός της παρούσας εργασίας ήταν η ανάπτυξη μεθόδου ανοσοχημικού προσδιορισμού των τριών μυκοτοξινών χωρίς τη χρήση ιχνηθετών με οπτικό αισθητήρα που βασίζεται στη φασματοσκοπία ανάκλασης λευκού φωτός. Ο αισθητήρας επιτρέπει την παρακολούθηση σε πραγματικό χρόνο βιομοριακών αντιδράσεων, μέσω της καταγραφής των μεταβολών του φάσματος συμβολής του φωτός που ανακλάται από ψηφίδα πυριτίου που φέρει υμένιο SiO2. O προσδιορισμός πολλαπλών αναλυτών επιτεύχθηκε με τη χρήση ψηφίδων που φέρουν περιοχές SiO2 διαφορετικού πάχους, σε καθεμία από τις οποίες ακινητοποιείται πρωτεϊνικό σύζευγμα διαφορετικής μυκοτοξίνης. Για τον προσδιορισμό διαβιβάζεται στη ψηφίδα, μίγμα προτύπου διαλύματος ή δείγματος δημητριακών (αραβοσίτου ή σιταριού), με διάλυμα που περιέχει τα ειδικά κατά των μυκοτοξινών αντισώματα και ακολουθεί διαβίβαση δευτέρου αντισώματος για την ενίσχυση του σήματος. Κατά τη διάρκεια της αντίδρασης, λαμβάνεται ένα φάσμα ανάκλασης από όλες τις περιοχές τις ψηφίδας και ειδικός αλγόριθμος προσδιορίζει το πάχος που αντιστοιχεί σε κάθε μία από αυτές, για την παρακολούθηση των αντιδράσεων. Η μέθοδος του ταυτόχρονου προσδιορισμού των τριών μυκοτοξινών ήταν ταχεία (12 min), με όρια ανίχνευσης 0.07, 1.25 και 1.0 ng/mL για AFB1, DON και FB1 αντίστοιχα, και δυναμικές περιοχές που εκτείνονταν έως τα 4.0, 250 και 50 ng/mL, αντίστοιχα. Λόγω απουσίας κινούμενων εξαρτημάτων, ο αναπτυχθείς αισθητήρας ταυτόχρονου προσδιορισμού πολλαπλών αναλυτών αναμένεται να αποτελέσει τη βάση φορητής συσκευής για τον επιτόπιο προσδιορισμό αναλυτών.Mycotoxins are chemical compounds produced by various fungal families that constitute a class of substances whose concentration should be determined in several food categories, but also in animal feed, due to the negative effects they have on human and animal health. The mycotoxins that are usually determined in food belong to three main categories: aflatoxins, fumonisins, and trichothecenes, from which the substances most frequently detected are aflatoxin B1 (AFM1), fumonisin B1 (FB1) and deoxinivalenol (DON). In this context, the aim of the present study was to develop a method for the simultaneous label-free immunochemical determination of the three mycotoxins implementing an optical sensor based on white light reflectance spectroscopy (WLRS). The sensor allows monitoring in real-time of biomolecular interactions by monitoring the changes in the interference spectrum reflected from a silicon chip modified with a silicon dioxide layer on which the reaction takes place. The determination of multiple analytes was accomplished by using chips with areas of varying thickness of silicon dioxide, each one of which has been biofunctionalized with a protein-conjugate of a different mycotoxin. For the assay, a mixture of calibrators or samples with a solution containing the anti-mycotoxin specific antibodies was run over the chip, followed by reaction with secondary for signal enhancement. A single reflection spectrum was received from all areas and it was deconvoluted into the spectrum of each individual area allowing the monitoring of the reactions taking place on it. The assay for the simultaneous determination of the three mycotoxins was fast (12 min) with detection limits 0.1, 1.25 and 1.0 ng/mL for AFB1, DON and FB1, respectively, and dynamic ranges extending to 5.0, 250 and 50 ng/mL, respectively. The sensor was applied to determine the three mycotoxins in cereal samples, mainly corn and wheat. Due to the lack of moving parts, the developed multi-analyte detection principle is expected to greatly facilitate the construction of a portable device determination of analytes at the point of need

The Wetting Behavior of Polymer Droplets: Effects of Droplet Size and Chain Length

Monte Carlo computer simulations were utilized to probe the behavior of homopolymer droplets adsorbed at solid surfaces as a function of the number of chains making up the droplets and varying droplet sizes. The wetting behavior is quantified via the ratio of the perpendicular to the parallel component of the effective radii of gyration of the droplets and is analyzed further in terms of the adsorption behavior of the polymer chains and the monomers that constitute the droplets. This analysis is complemented by an account of the shape of the droplets in terms of the principal moments of the radius of gyration tensor. Single-chain droplets are found to lie flatter and wet the substrate more than chemically identical multichain droplets, which attain a more globular shape and wet the substrate less. The simulation findings are in good agreement with atomic force microscopy experiments. The present investigation illustrates a marked dependence of wetting and adsorption on certain structural arrangements and proposes this dependence as a technique through which polymer wetting may be tuned

Unpublished Mediterranean and Black Sea records of marine alien, cryptogenic, and neonative species

To enrich spatio-temporal information on the distribution of alien, cryptogenic, and neonative species in the Mediterranean and the Black Sea, a collective effort by 173 marine scientists was made to provide unpublished records and make them open access to the scientific community. Through this effort, we collected and harmonized a dataset of 12,649 records. It includes 247 taxa, of which 217 are Animalia, 25 Plantae and 5 Chromista, from 23 countries surrounding the Mediterranean and the Black Sea. Chordata was the most abundant taxonomic group, followed by Arthropoda, Mollusca, and Annelida. In terms of species records, Siganus luridus, Siganus rivulatus, Saurida lessepsianus, Pterois miles, Upeneus moluccensis, Charybdis (Archias) longicollis, and Caulerpa cylindracea were the most numerous. The temporal distribution of the records ranges from 1973 to 2022, with 44% of the records in 2020–2021. Lethrinus borbonicus is reported for the first time in the Mediterranean Sea, while Pomatoschistus quagga, Caulerpa cylindracea, Grateloupia turuturu, and Misophria pallida are first records for the Black Sea; Kapraunia schneideri is recorded for the second time in the Mediterranean and for the first time in Israel; Prionospio depauperata and Pseudonereis anomala are reported for the first time from the Sea of Marmara. Many first country records are also included, namely: Amathia verticillata (Montenegro), Ampithoe valida (Italy), Antithamnion amphigeneum (Greece), Clavelina oblonga (Tunisia and Slovenia), Dendostrea cf. folium (Syria), Epinephelus fasciatus (Tunisia), Ganonema farinosum (Montenegro), Macrorhynchia philippina (Tunisia), Marenzelleria neglecta (Romania), Paratapes textilis (Tunisia), and Botrylloides diegensis (Tunisia).Stelios Katsanevakis, Michail Ragkousis, Maria Sini, Markos Digenis and Vasilis Gerovasileiou were supported by the Hellenic Foundation for Research and Innovation (HFRI) under the “First Call for HFRI Research Projects to support Faculty members and Researchers and the procurement of high-cost research equipment grant” (Project ALAS – “ALiens in the Aegean – a Sea under siege” (Katsanevakis et al. 2020b); Project Number: HFRI-FM17-1597). Konstantinos Tsirintanis was co-financed by Greece and the European Union (European Social Fund-ESF) through the Operational Programme “Human Resources Development, Education and Lifelong Learning”, 2014-2020, in the context of the Act “Enhancing Human Resources Research Potential by undertaking a Doctoral Research” Sub-action 2: IKY Scholarship Programme for PhD candidates in the Greek Universities. Maria Zotou was supported by the project “Coastal Environment Observatory and Risk Management in Island Regions AEGIS+” (MIS 5047038), implemented within the Operational Programme “Competitiveness, Entrepreneurship and Innovation” (NSRF 2014-2020), co financed by the Hellenic Government (Ministry of Development and Investments) and the European Union (European Regional Development Fund, Cohesion Fund). Razy Hoffman was supported by Yad-Hanadiv Foundation, through the Israel Society of Ecology and Environmental Sciences and Israel Nature and Parks Authority, an integrated program for establishing biological baselines and monitoring protocols for marine reserves in the Israeli Mediterranean Sea (Grant #10669). Tatiana Begun, Adrian Teaca and Mihaela Muresan were supported by the European Union’s Horizon 2020 BRIDGE-BS project under grant agreement no. 101000240. Fiona Tomas was supported by the project “Invasion of the tropical alga Halimeda incrassata in the Balearic Islands: ecology and invasion dynamics (AAEE119/2017)”, funded by the Vicepresidencia y Consejería de Innovación, Investigación y Turismo del Govern de les Illes Balears, with support from the European Union and FEDER funds, and the project “Una nueva alga invasora en el Mediterráneo: invasibilidad, detección y erradicación del alga tropical Halimeda incrassata (INVHALI)”, funded by the Fundación Biodiversidad, del Ministerio para la Transición Ecológica y el Reto Demográfico. Simonetta Fraschetti, Laura Tamburello, Antonia Chiarore were supported by the project PO FEAMP 2014-2020 - DRD n. 35/2019, “Innovazione, sviluppo e sostenibilità nel settore della pesca e dell'acquacoltura per la Regione Campania” (ISSPA 2.51) and the EU EASME - EMFF (Sustainable Blue Econ-omy) Project AFRIMED (http://afrimed-project.eu/, grant agreement N. 789059). Carlos Jimenez, Louis Hadjioannou, Vasilis Resaikos, Valentina Fossati, Magdalene Papatheodoulou, and Antonis Petrou were supported by MedPan Small Projects, Mava, and LIFE-IP. Louis Hadjioannou, Manos L. Moraitis and Neophytos Agrotis received funding from the European Union’s Horizon 2020 research and innovation program within the framework of the CMMI/MaRITeC-X project under grant agreement No. 857586. Ernesto Azzurro was supported by the project USEIt - Utilizzo di Sinergie operative per la gestione integrata specie aliene Invasive in Italia, funded by the research programme @CNR. Antonietta Rosso and Francesco Sciuto were supported by the University of Catania through “PiaCeRi-Piano Incentivi per la Ricerca di Ateneo 2020–22 linea di intervento 2.” This is the Catania Paleoecological Research Group contribution n. 484. Diego K. Kersting was supported by the Beatriu de Pinós programme funded by the Secretary of Universities and Research (Government of Catalonia) and the Horizon 2020 programme of research and innovation of the European Union under the Marie Sklodowska-Curie grant agreement No 801370. Francesco Tiralongo was supported by the AlienFish project of Ente Fauna Marina Mediterranea (Scientific Organization for Research and Conservation of Marine Biodiversity, 96012 Avola, Italy), a citizen science project for monitoring and studying rare and non-indigenous fish in Italian waters. Adriana Vella, was supported by funds through the BioCon_Innovate Research Excellence Grant from the University of Malta awarded to her. Noel Vella was supported by REACH HIGH Scholars Programme-Post Doctoral Grant for the FINS project. Some of the records provided by Victor Surugiu were obtained during surveys carried out within the framework of the project “Adequate management of invasive species in Romania, in accordance with EU Regulation 1143/2014 on the prevention and management of the introduction and spread of invasive alien species”, SMIS 2014+ 120008, coordinated by the Romanian Ministry of Environment, Water and Forests in partnership with the University of Bucharest (2018–2022). Alan Deidun and Alessio Marrone were supported by the “Spot The Alien” citizen science campaign for the monitoring of the Alien species in the Maltese archipelago and by the Interreg Italia-Malta Harmony project. The authors from the National Institute of Biology (Slovenia) acknowledge the financial support of the Slovenian Research Agency (Research Core Funding No. P1-0237) and of the Ministry of Agriculture, Forestry and Food (project “Survey of the species richness and abundance of alien species in the Slovenian Sea”). Emanuele Mancini and Fabio Collepardo Coccia were supported by the project PO-FEAMP 2014-2020 “BIOBLITZ: research, knowledge and participation for the sustainable management of marine resources (BioBlitz Blu 2020)” coordinated by CURSA for MIPAAF, the Italian Ministry of Agricultural, Food and Forestry Policies, Measure 1.40 - Protection and restoration of biodiversity and marine ecosystems and compensation schemes in the context of sustainable fishing activities. Daniele Grech was supported by the PO-FEAMP 2014-2020 project ECOGESTOCK “Approccio ECOsistemico per la tutela e la GEStione delle risorse biologiche e STOCK ittici nelle acque interne”, the citizen science project Progetto Fucales: chi le ha viste? and the Paralenz Every dive counts sponsor. Jamila Rizgalla was supported by the project Snowball for the monitoring of alien species in Libyan waters له اهتفش له اهتدطصا ؟) have you seen it have you fished it?). Gerasimos Kondylatos and Dimitrios Mavrouleas were supported by the project “EXPLIAS” (MIS (ΟΠΣ): 5049912), design and piloting methods of commercial exploitation of invasive alien species with a view to contributing to their population control, coordinated by the National Technical University of Athens with the collaboration of the Hellenic Centre for Marine Research and the University of the Aegean and co-founded by Greece and the European Union. G. Kondylatos and Savvas Nikolidakis were supported by the project “SAMOS” (ID CODE: 32.2072004/001), a study for a submarine productive park in Marathokampos of Samos. Paraskevi K. Karachle, Aikaterini Dogrammatzi, Giorgos A. Apostolopoulos, Kassiani Konida and Melina Nalmpanti were supported by the project “4ALIEN: Biology and the potential economic exploitation of four alien species in the Hellenic Seas”, funded by NRSF 2017-2020 (MIS (ΟΠΣ): 5049511). Fabio Crocetta and Riccardo Virgili were partially funded by the project PO FEAMP Campania 2014–2020, DRD n. 35 of 15th March 2018, Innovazione, sviluppo e sostenibilità nel settore della pesca e dell’acquacoltura per la regione Campania, Misura 2.51, WP5, Task 5.5 Presenza e distribuzione di specie non indigene del macrozoobenthos e del necton in Campania. Michel Bariche was partially funded by the University Research Board of the American University of Beirut (DDF 103951/2592). Constantinos G. Georgiadis, Dimitra Lida Rammou, Paschalis Papadamakis and Sotiris Orfanidis were supported by the MSFD monitoring program. Sonia Smeraldo was supported by the MPA-Engage project, led by the Institute of Marine Sciences of the Spanish National Research Council and funded by the Interreg MED program. Evgeniia Karpova acknowledge that the publication of this article was in part carried out within the framework of the state assignment of the FRC IBSS “Patterns of Formation and Anthropogenic Transformation of Biodiversity and Bioresources of the Azov– Black Sea Basin and Other Regions of the World Ocean” (No. 121030100028-0). Elena Slynko’s work was carried out within the framework of a State Assignment no. 121051100109-1 of IBIW RAS. Manuela Falautano and Luca Castriota were supported by ISPRA citizen science campaigns for the monitoring of alien species through the dedicated institutional project ([email protected]). María Altamirano was supported by the project RUGULOPTERYX funded by Fundación Biodiversidad-Ministerio para la Transición Ecológica y el reto Demográfico (Spain) and the project UMA20-FEDERJA-006 with support from the European Union and FEDER funds and Junta de Andalucía. Records provided by L. Mangialajo were collected in the framework of projects funded by the Pew Charitable Trust, by the European Commission (AFRIMED, http://afrimed-project.eu/, grant agreement N. 789059) and by the Académie 3 de l’Université Côte d’Azur (projet CONVOST).Peer reviewe

Unpublished Mediterranean and Black Sea records of marine alien, cryptogenic, and neonative species

To enrich spatio-temporal information on the distribution of alien, cryptogenic, and neonative species in the Mediterranean and the Black Sea, a collective effort by 173 marine scientists was made to provide unpublished records and make them open access to the scientific community. Through this effort, we collected and harmonized a dataset of 12,649 records. It includes 247 taxa, of which 217 are Animalia, 25 Plantae and 5 Chromista, from 23 countries surrounding the Mediterranean and the Black Sea. Chordata was the most abundant taxonomic group, followed by Arthropoda, Mollusca, and Annelida. In terms of species records, Siganus luridus, Siganus rivulatus, Saurida lessepsianus, Pterois miles, Upeneus moluccensis, Charybdis (Archias) longicollis, and Caulerpa cylindracea were the most numerous. The temporal distribution of the records ranges from 1973 to 2022, with 44% of the records in 2020–2021. Lethrinus borbonicus is reported for the first time in the Mediterranean Sea, while Pomatoschistus quagga, Caulerpa cylindracea, Grateloupia turuturu, and Misophria pallida are first records for the Black Sea; Kapraunia schneideri is recorded for the second time in the Mediterranean and for the first time in Israel; Prionospio depauperata and Pseudonereis anomala are reported for the first time from the Sea of Marmara. Many first country records are also included, namely: Amathia verticillata (Montenegro), Ampithoe valida (Italy), Antithamnion amphigeneum (Greece), Clavelina oblonga (Tunisia and Slovenia), Dendostrea cf. folium (Syria), Epinephelus fasciatus (Tunisia), Ganonema farinosum (Montenegro), Macrorhynchia philippina (Tunisia), Marenzelleria neglecta (Romania), Paratapes textilis (Tunisia), and Botrylloides diegensis (Tunisia).peer-reviewe

Development of biosensors for multiplexed mycotoxins determination

Mycotoxins are chemical compounds produced by various fungal families that constitute a class of substances whose concentration should be determined in several food categories, but also in animal feed, due to the negative effects they have on human and animal health. The mycotoxins that are usually determined in food belong to three main categories: aflatoxins, fumonisins, and trichothecenes, from which the substances most frequently detected are aflatoxin B1 (AFM1), fumonisin B1 (FB1) and deoxinivalenol (DON). In this context, the aim of the present study was to develop a method for the simultaneous label-free immunochemical determination of the three mycotoxins implementing an optical sensor based on white light reflectance spectroscopy (WLRS). The sensor allows monitoring in real-time of biomolecular interactions by monitoring the changes in the interference spectrum reflected from a silicon chip modified with a silicon dioxide layer on which the reaction takes place. The determination of multiple analytes was accomplished by using chips with areas of varying thickness of silicon dioxide, each one of which has been biofunctionalized with a protein-conjugate of a different mycotoxin. For the assay, a mixture of calibrators or samples with a solution containing the anti-mycotoxin specific antibodies was run over the chip, followed by reaction with secondary for signal enhancement. A single reflection spectrum was received from all areas and it was deconvoluted into the spectrum of each individual area allowing the monitoring of the reactions taking place on it. The assay for the simultaneous determination of the three mycotoxins was fast (12 min) with detection limits 0.1, 1.25 and 1.0 ng/mL for AFB1, DON and FB1, respectively, and dynamic ranges extending to 5.0, 250 and 50 ng/mL, respectively. The sensor was applied to determine the three mycotoxins in cereal samples, mainly corn and wheat. Due to the lack of moving parts, the developed multi-analyte detection principle is expected to greatly facilitate the construction of a portable device determination of analytes at the point of need.Οι μυκοτοξίνες είναι χημικές ενώσεις που παράγονται από διάφορες οικογένειες μυκήτων των οποίων η συγκέντρωση επιβάλλεται να προσδιορίζεται σε αρκετές κατηγορίες τροφίμων, λόγω των αρνητικών επιπτώσεων που έχουν στην υγεία των ανθρώπων και των ζώων. Οι μυκοτοξίνες που συνήθως ανιχνεύονται στα τρόφιμα ανήκουν κυρίως σε τρεις κατηγορίες: αφλατοξίνες, φουμονισίνες, και τριχοθηκίνες, με κύριους εκπροσώπους αυτών την αφλατοξίνη Β1 (AFB1), τη φουμονισίνη Β1 (FB1) και τη δεοξυνιβαλενόλη (DON). Στο πλαίσιο αυτό, σκοπός της παρούσας εργασίας ήταν η ανάπτυξη μεθόδου ανοσοχημικού προσδιορισμού των τριών μυκοτοξινών χωρίς τη χρήση ιχνηθετών με οπτικό αισθητήρα που βασίζεται στη φασματοσκοπία ανάκλασης λευκού φωτός. Ο αισθητήρας επιτρέπει την παρακολούθηση σε πραγματικό χρόνο βιομοριακών αντιδράσεων, μέσω της καταγραφής των μεταβολών του φάσματος συμβολής του φωτός που ανακλάται από ψηφίδα πυριτίου που φέρει υμένιο SiO2. O προσδιορισμός πολλαπλών αναλυτών επιτεύχθηκε με τη χρήση ψηφίδων που φέρουν περιοχές SiO2 διαφορετικού πάχους, σε καθεμία από τις οποίες ακινητοποιείται πρωτεϊνικό σύζευγμα διαφορετικής μυκοτοξίνης. Για τον προσδιορισμό διαβιβάζεται στη ψηφίδα, μίγμα προτύπου διαλύματος ή δείγματος δημητριακών (αραβοσίτου ή σιταριού), με διάλυμα που περιέχει τα ειδικά κατά των μυκοτοξινών αντισώματα και ακολουθεί διαβίβαση δευτέρου αντισώματος για την ενίσχυση του σήματος. Κατά τη διάρκεια της αντίδρασης, λαμβάνεται ένα φάσμα ανάκλασης από όλες τις περιοχές τις ψηφίδας και ειδικός αλγόριθμος προσδιορίζει το πάχος που αντιστοιχεί σε κάθε μία από αυτές, για την παρακολούθηση των αντιδράσεων. Η μέθοδος του ταυτόχρονου προσδιορισμού των τριών μυκοτοξινών ήταν ταχεία (12 min), με όρια ανίχνευσης 0.07, 1.25 και 1.0 ng/mL για AFB1, DON και FB1 αντίστοιχα, και δυναμικές περιοχές που εκτείνονταν έως τα 4.0, 250 και 50 ng/mL, αντίστοιχα. Λόγω απουσίας κινούμενων εξαρτημάτων, ο αναπτυχθείς αισθητήρας ταυτόχρονου προσδιορισμού πολλαπλών αναλυτών αναμένεται να αποτελέσει τη βάση φορητής συσκευής για τον επιτόπιο προσδιορισμό αναλυτών

Fast Deoxynivalenol Determination in Cereals Using a White Light Reflectance Spectroscopy Immunosensor

Deoxynivalenol (DON) is a mycotoxin produced by certain Fusarium species and found in a high percentage of wheat and maize grains cultured worldwide. Although not so toxic as other mycotoxins, it exhibits both chronic and acute toxicity, and therefore methods for its fast and accurate on-site determination are highly desirable. In the current work, we employ an optical immunosensor based on White Light Reflectance Spectroscopy (WLRS) for the fast and sensitive immunochemical label-free determination of DON in wheat and maize samples. The assay is completed in 12 min and has a quantification limit of 2.5 ng/mL in buffer corresponding to 125 μg/kg in whole grain which is lower than the maximum allowable concentrations set by the regulatory authorities for grains intended for human consumption. Several extraction protocols have been compared, and the highest recovery (>90%) was achieved employing distilled water. In addition, identical calibration curves were received in buffer and wheat/maize extraction matrix providing the ability to analyze the grain samples using calibrators in buffer. Recoveries of DON from spiked wheat and maize grain samples ranged from 92.0(±4.0) to 105(±4.0)%. The analytical performance of the WLRS immunosensor, combined with the short analysis time and instrument portability, supports its potential for on-site determinations

Fast Deoxynivalenol Determination in Cereals Using a White Light Reflectance Spectroscopy Immunosensor

Deoxynivalenol (DON) is a mycotoxin produced by certain Fusarium species and found in a high percentage of wheat and maize grains cultured worldwide. Although not so toxic as other mycotoxins, it exhibits both chronic and acute toxicity, and therefore methods for its fast and accurate on-site determination are highly desirable. In the current work, we employ an optical immunosensor based on White Light Reflectance Spectroscopy (WLRS) for the fast and sensitive immunochemical label-free determination of DON in wheat and maize samples. The assay is completed in 12 min and has a quantification limit of 2.5 ng/mL in buffer corresponding to 125 mu g/kg in whole grain which is lower than the maximum allowable concentrations set by the regulatory authorities for grains intended for human consumption. Several extraction protocols have been compared, and the highest recovery (>90%) was achieved employing distilled water. In addition, identical calibration curves were received in buffer and wheat/maize extraction matrix providing the ability to analyze the grain samples using calibrators in buffer. Recoveries of DON from spiked wheat and maize grain samples ranged from 92.0(+/- 4.0) to 105(+/- 4.0)%. The analytical performance of the WLRS immunosensor, combined with the short analysis time and instrument portability, supports its potential for on-site determinations

Partial epididymal rupture and spermatic cord haematoma with an associated secondary testicular torsion due to blunt scrotal injury, in a 12-year-old boy

Scrotal injuries are not very common in children and are mostly due to blunt trauma from direct injury, sports injuries or motor vehicle accidents. Traumatic testicular torsion in children has been also infrequently reported in the literature. To ensure testicular salvage, an urgent and specialised diagnosis and management are necessary. We present a case of a partial epididymal rupture and spermatic cord haematoma with an associated secondary testicular torsion due to blunt scrotal injury, in a 12-year-old boy

Testis-Sparing Surgery for Non-Palpable Leydig Cell Tumors in Prepubertal Children

Leydig cell tumor (LCT) is an infrequent stromal neoplasm of the testis with an incidence of less than 3% of all gonadal tumors in the general male population. Only 25% is found in prepubertal children, where Leydig cell tumors are always reported benign. The hospital records of two prepubertal male children, who underwent organ sparing surgery for testicular LCT the last five years, were retrospectively reviewed. In both of them, the lesion was incidentally found during a scrotal ultrasonography for testicular pain. The diagnosis of a benign LCT was based on the pre-operative physical examination and imaging (Ultrasound-US, Magnetic Resonance Imaging-MRI) as well as the negative tumor markers. A testicle-sparing procedure was decided and the pathologic examination of the surgical specimen confirmed the diagnosis. No tumor recurrence was noted on follow-up. Testis-sparing surgery provides the possibility of complete excision of such lesions and should be considered as the treatment of choice