718 research outputs found

    Use of FTA® classic cards for epigenetic analysis of sperm DNA

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    FTA® technologies provide the most reliable method for DNA extraction. Although FTA technologies have been widely used for genetic analysis, there is no literature on their use for epigenetic analysis yet. We present for the first time, a simple method for quantitative methylation assessment based on sperm cells stored on Whatman FTA classic cards. Specifically, elution of seminal DNA from FTA classic cards was successfully tested with an elution buffer and an incubation step in a thermocycler. The eluted DNA was bisulfite converted, amplified by PCR, and a region of interest was pyrosequenced

    On the masked mycotoxin zearalenone-14-glucoside. Does the mask truly hide?

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    In the matter of foodborne mycotoxins, beside a number of regulated compounds, regulations are totally missing for phase-II plant metabolites - the toxicological knowledge of which is still in its infancy. Currently, zearalenone-14-glucoside is in the pipeline and its toxicological role is under a glowing scientific debate. In our work it clearly showed high toxicological concerns as it is prone to conversion to well-known toxic compounds (i.e. zearalenone and both zearalenol isomers) when exposed to breast cancer cells culture. The need of future risk assessment studies has been pointed out accordingly

    Methy-sens Comet assay and DNMTs transcriptional analysis as a combined approach in epigenotoxicology

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    Epigenotoxicology needs simple and fast tools to assess xenobiotic epigenetic load. This work proposes a comet assay modification designed to detect global methylation changes (Methysens Comet) through enzymatic digestion with two restriction enzymes (HpaII, MspI). In the methylation-sensitive protocol tested for repeatability on A549 cells, nickel chloride induced hypermethylation and decitabine-induced hypomethylation. A concomitant assessment of DNA methyltransferases (DNMTs) genes transcriptional levels has been performed, to implement a multifunctional approach to epigenotoxicology. Methy-sens Comet showed a general good repeatability and sensitivity to methylation changes while DNMTs transcriptional levels granted additional proof of xenobiotic-induced impairment of methylome maintenance

    What does egg size tell us about hatching phenology in Heterocypris incongruens?

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    The timing of many phenological events (e.g. egg hatching, development and reproduction) is the result of the interaction of genotype, environmental factors and maternal effects. In many taxa it is known that egg size and development time are linked and that, especially in temporary habitats, mothers, anticipating the environment that their offspring will face, can determine egg size (and its variability) to spread the risk connected with environmental unpredictability. We investigated this phenomenon in a widespread microcrustacean: Heterocypris incongruens. We measured the size of eggs produced under laboratory-controlled conditions (24 °C and two photoperiods) by genetically identical females of an ecotype typical of Northern Italian vernal pools. We used 12:12 L:D as a proxy of incoming favourable but unpredictable winter-spring conditions in the Po river plain and 16:8 L:D as a proxy of incoming dry, predictable but unfavourable conditions (that we expected to induce resting egg production). The lack of genetic variation makes it easy to investigate the genetic basis of the effect of the photoperiod experienced by the mother on egg size. Mean diameter was larger in eggs produced at 12:12 L:D (N = 151, 133.16 μm) than at 16:8 L:D (N = 106 129.93 μm). Eggs that did not hatch within 100 days from deposition were classified as resting eggs: their percentage increased from 48% at 12:12 L:D to 73% at 16:8 L:D. Egg diameter at deposition was not significantly larger in non-resting (N = 42, 133.45 μm) than in resting eggs (N = 68, 130.63 μm). Development time of non-resting eggs was highly variable (from 2 to 100 days), it was not related to egg size at deposition and did not change with photoperiod. Any possible effect of egg size at deposition on development time was probably masked by the high variability of the latter variable. During our observation time (every 24 hours for at least 100 days), egg diameter of both resting and non-resting eggs (for a total of 1841 measurements) increased following an asymptotic model. Size increment was slower and eventually larger in eggs laid at 16:8 L:D than at 12:12 L:D and was probably linked to embryo growth in both resting and non-resting eggs. Final diameter of both resting and non-resting eggs did not show a threshold size and its variability might be linked to the hatching asynchrony observed in resting and non-resting eggs. Our work contributes to the budding field of egg focused mother controlled phenology

    The Pottesman Collection in the British Museum. Early Dynastic and Sargonic administrative texts. With an Appendix on a Palmyrene Inscription

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    Edizione, trascrizione, traduzione e commento di un frammento di iscrizione palmirena inedita presente nella collezione Pottesman del British Museum (Appendice Agostini).The British Museum houses a small collection of six cuneiform tablets and a Palmyrene dedicatory inscription purchased in 1987 from the private collection of Solomon Pottesman. The aim of the present contribution is to provide a catalog of this lot and an edition of the so far unpublished cuneiform texts. In the appendix, Alessio Agostini added the edition of the Palmyrene inscription, which would have otherwise gone beyond the capabilities of the present author

    Recensione di Cecilia Falchini (2023). Ruperto di deutz - Un’intima familiarità. Antologia, Edizioni Qiqajon (Comunità di Bose), Magnano (Bi), 281 pp.

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    Review of Cecilia Falchini (2023). Ruperto di deutz - Un’intima familiarità. Antologia, Edizioni Qiqajon (Comunità di Bose), Magnano (Bi), 281 pp. Author: Alessio MagogaRecensione di Cecilia Falchini (2023). Ruperto di deutz - Un’intima familiarità. Antologia, Edizioni Qiqajon (Comunità di Bose), Magnano (Bi), 281 pp. Autor: Alessio Magog

    What does egg size tell us abiut development and hatching time in Heterocypris incongruens?

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    Colonisation of ephemeral ponds requires adaptations in life history and in hatching phenology: organisms must take advantage of ponds filling and “hedge their bets” for the possibility to complete their life cycle and produce resting stages that ensure that the population will not go extinct. The timing of many phenological events (e.g. egg hatching, beginning of development phases and reproduction) results from a complex interplay among organism genotype, environmental factors (e.g. temperature and photoperiod) and maternal effects. The relationship between egg size and development time is known and it is generally accepted that larger eggs take longer to develop than smaller ones. The production of eggs with variable size may represent a strategy by which a mother spread the risk connected with life in a temporary habitat. In laboratory experiments carried out at 24 °C, we evaluated the effect of egg size on egg development time in a clonal lineage of Heterocypris incongruens typical of Northern Italy vernal pools. The lack of genetic variation expected among clonal organisms make them ideal material for investigating maternal effect (e.g. the environment the mother experienced). We measured the size of 50 eggs released at 12:12 L:D photoperiod, chosen as a proxy of favourable but unpredictable late winter-spring conditions, and 62 eggs at 16:8 L:D photoperiod, proxy of incoming a dry predictably unfavourable season inducing resting egg production. Each egg was identified, photo recorded, measured at different times from deposition for at least 200 days, for a total of 1841 measurements. At deposition, mean diameter was smaller in eggs released at 16:8 L:D (129.9 μm sd 7.69) than at 12:12L:D (133.1 μm sd 8.46) and, as expected, hatching percentage decreased from 52% at 12:12 L:D to 27% at 16:8 L:D. Egg diameter did not differ between resting (130.4 μm sd 8.13) and non resting eggs (133.0 μm sd 8.09) and did not affect hatching time that was highly variable (from 2 to 100 days) but did not vary with photoperiod. During our observation time, egg diameter increased 1.09-1.14 times following an asymptotic model. Size increment was higher (and slower) in eggs produced at 16:8 L:D than in eggs produced at 12:12 L:D. It was highly variable especially in resting eggs produced at 12:12 L:D, and was linked to the initial growth that occurred within 0-3 days from deposition. Our preliminary data show that, in H. incongruens, the egg size at deposition is affected by the environment the mother experienced, hatching time is not affected by egg size at deposition and early embryo developmental process occurs in resting eggs whose number of cells is about one hundred

    Handheld-Impedance-Measurement System with seven-decade capability and potentiostatic function

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    This paper describes design and test of a new impedance-measurement system for nonlinear devices that exhibits a seven-decade range and works down to a frequency of 0.01 Hz. The system is specifically designed for electrochemical measurements, but the proposed architecture can be employed in many other fields where flexible signal generation and analysis are required. The system employs an unconventional signal generator based on two pulsewidth modulation (PWM) oscillators and an autocalibration system that allows uncertainties of less than 3% to be obtained over a range of 1 kΩ to 100 GΩ. A synchronous demodulation processing allows the noise superimposed to the low-amplitude input signals to be made negligibl

    VALUTAZIONE DEL DANNO PRIMARIO ED OSSIDATIVO AL DNA IN LINFOCITI SALIVARI PER IL MONITORAGGIO DEGLI EFFETTI BIOLOGICI PRECOCI CAUSATI DALL’INQUINAMENTO ATMOSFERICO NEI BAMBINI: STUDIO MAPEC

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    VALUTAZIONE DEL DANNO PRIMARIO ED OSSIDATIVO AL DNA IN LINFOCITI SALIVARI PER IL MONITORAGGIO DEGLI EFFETTI BIOLOGICI PRECOCI CAUSATI DALL’INQUINAMENTO ATMOSFERICO NEI BAMBINI: STUDIO MAPEC Samuele Vannini (1) - Sara Levorato (1) - Elisabetta Ceretti (2) - Milena Villarini (1) - Silvia Bonetta (3) - Cristina Fatigoni(1) - Annalaura Carducci (4) - Tania Salvatori (1) - Maria Rosaria Tumolo (5) - Alessio Perotti (6) - Silvia Bonizzoni (7) - Alberto Bonetti (8) - Massimo Moretti (1) - Umberto Gelatti (2) Dipartimento di Scienze Farmaceutiche, Università degli Studi di Perugia, Perugia, Italia (1) - Dipartimento di Specialità Medico-Chirurgiche, Scienze Radiologiche e Sanità Pubblica, Università degli Studi di Brescia, Brescia, Italia (2) - Dipartimento di Scienze della Sanità Pubblica e Pediatriche, Università degli Studi di Torino, Torino, Italia (3) - Dipartimento di Biologia, Università degli Studi di Pisa, Pisa, Italia (4) - Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento, Lecce, Italia (5) - Dipartimento di Bioscienze, Università del Studi di Parma, Parma, Italia (6) - Ufficio Minori, Comune di Brescia, Brescia, Italia (7) - Centro Servizi Multisettoriale e Tecnologico, Csmt Gestione S.c.a.r.l., Brescia, Italia (8) Il particolato è l’inquinante atmosferico che provoca i maggiori danni alla salute umana in Europa (Agenzia Europea dell’Ambiente; www.eea.europa.eu). Tra le prime 30 città più inquinate del continente, oltre la metà sono italiane, in particolare quelle situate nella pianura Padana, con concentrazioni di PM10, PM2,5 e NOx ben al di sopra dei valori di riferimento europei. Studi epidemiologici hanno rilevato un’associazione positiva tra esposizione ad inquinamento atmosferico, soprattutto al PM, ed incidenza e mortalità per diverse malattie cronico-degenerative, come cancro al polmone, malattie cardiovascolari e diabete. I bambini, in particolar modo, risultano esposti ad un elevato rischio per quanto riguarda gli effetti a breve e lungo termine dell’inquinamento atmosferico. Dati recenti suggeriscono che alcune alterazioni genetiche che si verificano nella prima infanzia posso incrementare il rischio di malattie cronico-degenerative in età adulta. L’obiettivo dello studio MAPEC (Monitoring Air Pollution Effects on Children for supporting Public Health policy) è quello di valutare l’associazione tra inquinamento atmosferico e biomarcatori di effetti genotossici precoci nei bambini, e di proporre un modello per la stima del rischio globale causato dagli inquinanti atmosferici. Lo studio ha previsto il reclutamento di circa 1.000 bambini, di età compresa tra 6-8 anni, in cinque città italiane (200 bambini per città) caratterizzate da diverse concentrazioni di inquinanti atmosferici: Brescia, Torino, Pisa, Perugia e Lecce. Campioni di saliva sono stati raccolti in due differenti stagioni, inverno e primavera, caratterizzate da differenze qualitative e quantitative degli inquinanti atmosferici (1.000 × 2 = 2.000 campioni). Nei linfociti salivari è stato valutato il danno primario e ossidativo al DNA mediante il test della microgel elettroforesi su singole cellule (comet assay). Alla conferenza verranno presentati i risultati preliminari relativi al campionamento invernale
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