Cytophotometric Analyses and in Vitro Culture Test in the Embryo First Node of Old Triticum Durum Caryopses

SUMMARYThe nuclei of the first node embryo of Triticum durum show the same amount of histone bound to DNA in young and in aged seeds in line with earlier findings of remaining metabolic activity. Furthermore the explants of the first node from the old seeds analyzed, are still able to generate calli and plantlets and might be usefull to recover embryos from aged seeds

A modified culture medium for improved isolation of marine vibrios

Marine Vibrio members are of great interest for both ecological and biotechnological research, which often relies on their isolation. Whereas many efforts have been made for the detection of food-borne pathogenic species, much less is known about the performances of standard culture media toward environmental vibrios. We show that the isolation/enumeration of marine vibrios using thiosulfate-citrate-bile salts-sucrose agar (TCBS) as selective medium may be hampered by the variable adaptability of different taxa to the medium, which may result even in isolation failure and/or in substantial total count underestimation. We propose a modified TCBS as isolation medium, adjusted for marine vibrios requirements, which greatly improved their recovery in dilution plate counts, compared with the standard medium. The modified medium offers substantial advantages over TCBS, providing more accurate and likely estimations of the actual presence of vibrios. Modified TCBS allowed the recovery of otherwise undetected vibrios, some of which producing biotechnologically valuable enzymes, thus expanding the isolation power toward potentially new enzyme-producers Vibrio taxa. Moreover, we report a newly designed Vibrio-specific PCR primers pair, targeting a unique rpoD sequence, useful for rapid confirmation of isolates as Vibrio members and subsequent genetic analyses

Fat plug myringoplasty: Analysis of a safe procedure for small tympanic perforations

Objective: Assessment of value of fat plug myringoplasty as office-based procedure in small tympanic membrane perforations. Material and Methods: Prospective study of consecutive patients undergoing fat plug myringoplasty under local anesthesia. Minimum follow-up considered was 6 months, which included several microscopic examinations. All data were recorded and analyzed statistically. Results: 33 patients matched the inclusion criteria; the mean perforation size was 2.8 mm. No change in PTA was noted after surgery in the whole series. Recurrence of perforation was detected in 7 cases (21%), which was statistically related with the cause of perforation rather than location or size (p-value <0.01). Conclusions: Fat plug myringoplasty is a safe procedure with very low rate of complications, and it is an office-based procedure well tolerate by all patients. Results in case of primary surgery are excellent, with a high success rate (90%). In case of recurrent perforation after traditional surgery, success rate get decreased

Effect of Metal Cocatalysts and Operating Conditions on the Product Distribution and the Productivity of the CO2 Photoreduction

open7The CO2 photoreduction is a promising way to convert one of the most abundant greenhouse gases to valuable chemicals. The photoreduction in the liquid phase is limited by the low solubility of CO2 in water, but this point is overcome here by using an innovative photoreactor, which allows one to work up to pressures of 20 bar, improving the overall productivity. The photoreduction was performed in the presence of Na2SO3 and using in primis commercial titanium dioxide (P25) and a set of titania catalysts functionalized by surface deposition of either monometallic or bimetallic cocatalysts. The gaseous products were hydrogen and traces of CO, while, in the liquid phase, formic acid/formate, formaldehyde and methanol were quantitatively detected. The pH was observed to shift the products distribution. A neutral environment led mainly to hydrogen and methanol, while, at pH 14, formate was the most abundant compound. The trend for monometallic cocatalysts showed enhanced productivity when using noble metals (i.e., gold and platinum). In order to limit the cost of the catalytic material, bimetallic cocatalysts were explored, adding titania with Au+Ag or Au+Pt. This may open to the possibility of performing the reaction with a smaller amount of the most expensive metals. In the end, we have expressed some conclusions on the cost of the photocatalysts here employed, to support the overall feasibility assessment of the process.openConte F.; Villa A.; Prati L.; Pirola C.; Bennici S.; Ramis G.; Rossetti I.Conte, F.; Villa, A.; Prati, L.; Pirola, C.; Bennici, S.; Ramis, G.; Rossetti, I

Canal switch and re-entry phenomenon in benign paroxysmal positional vertigo: difference between immediate and delayed occurrence

Studio prospettico ideato per la valutazione delle differenze tra la conversione canalare o il rientro degli otoliti nei canali semicircolari successivo alle manovre terapeutiche nei pazienti affetti da VPPB. Sono stati valutati 196 pazienti affetti da VPPB, 127 dei quali corrispondevano ai criteri di inclusione. L’età media dei pazienti era di 54.74 anni. Il canale orizzontale è stato coinvolto in 30 casi e il canale posteriore in 97 pazienti. I pazienti con sordità neurosensoriale presentavano forme ricorrenti di VPPB, rispetto a quelli con udito normale. L’immediato rientro canalare è stato diagnosticato in 3 pazienti con VPPB del canale laterale, tutti con nistagmo geotropo. 7 pazienti con VPPB del canale posteriore hanno presentato un rientro canalare immediato e 5 la forma ritardata. I pazienti con rientro canalare ritardato avevano precedentemente subito più di 2 manovre di riposizionamento. Il rientro canalare non è risultato connesso al tipo di manovra eseguita. Il tempo di attesa tra l’esecuzione della manovra liberatoria e il test di verifica si è rivelato importante ai fini del rientro canalare immediato. La recidiva della BPPV dopo un mese dalle manovre liberatorie si è riscontrata in 20 pazienti ed è stata più frequente in quei pazienti che hanno avuto un fenomeno di rientro canalare. La conversione canalare ed il fenomeno del rientro canalare rappresentano delle entità cliniche che devono essere considerate dal medico che tratta le VPPB. Appare importante distinguere un rientro da un fallimento della manovra in caso di forme immediate, o da una recidiva di patologia in caso di forme ritardate. L’esecuzione del test di verifica del successo terapeutico dopo manovre di riposizionamento deve avere un distacco temporale sufficientemente ampio al fine di evitare il reflusso immediato di otoliti nei canali.This prospective study was designed to evaluate the differences between immediate and delayed canal re-entry of otoliths after therapeutic manoeuvres in patients with benign paroxysmal positional vertigo (BPPV). A total of 196 patients with BPPV were visited and 127 matched our inclusion criteria. The mean age was 54.74 years. The horizontal semicircular canal (HSC) was involved in 30 cases and the posterior semicircular canal (PSC) in 97 patients. Patients with hearing loss in the ear affected by BPPV have a more recurrent form, compared to those with normal hearing. An immediate canal re-entry was recorded in 3 patients with HSC BPPV, all with geotropic nystagmus. In 7 patients with PSC BPPV, the immediate canal re-entry was detected and the delayed form was noted in 5 patients. The patients with the delayed canal re-entry underwent more than 2 previous manoeuvres. The canal re-entry was not related to the manoeuvre performed. The timing of the Dix-Hallpike test to verify the resolution of the BPPV had a significant role in immediate canal re-entry. A recurrence in the follow-up at least one month after treatment was recorded in 20 patients and was more frequent in patients that had canal re-entry. The canal re-entry or canal switch is a clinical entity that should be kept in mind of the neurotologist when approaching BPPV patients. It is important to distinguish it from recurrence when delayed and from manoeuvre failure when immediate. The timing of manoeuvre performing, in particular the final verification test after therapeutic sessions, is important to prevent the immediate reflux of particles into canals

Esperimento pilota mirato all’analisi dell’effetto della stimolazione immunitaria su A. viridis, finalizzato alla produzione di composti biologicamente attivi

L’esperimento mira a valutare l’eventuale attività antibatterica di peptidi prodotti da individui di Anemonia viridis in seguito a stimolazione immunitaria e quindi alla realizzazione di un protocollo riproducibile e ottimizzato per la produzione di tali composti

Applicazione di un protocollo di immuno-stimolazione su individui di Pracambarus clarkii mirato alla produzione di peptidi con attività biologica

Uno dei principali problemi inerenti l’identificazione e l’analisi di nuovi peptidi naturali di origine marina è la disponibilità in termini quantitativi del materiale bioattivo (Cragg et al., 1997). Infatti, le concentrazioni di tali peptidi negli invertebrati marini sono spesso meno del 10-6 % del peso umido, ed il loro rendimento a seguito delle procedure di estrazione tradizionali risulta ancora più basso. Tuttavia, sono molteplici i fattori che possono influenzare l'esito della identificazione degli Anti Microbial Peptides (AMP), come le variazioni geografiche e stagionali (sito di campionamento), le diverse fasi della vita (specie pelagiche o bentoniche), l'età, il sesso e lo stato fisiologico (malattia, allevamento, muta). A causa della difficoltà nell’ottenere quantità sufficienti di composti bioattivi, potrebbe essere più produttivo dissezionare in diversi tessuti/organi gli invertebrati selezionati per poi estrarre il materiale da saggiare (Haug et al., 2002a, 2002b, 2004) ciò al fine di separare i peptidi di interesse da migliaia di sostanze inattive. Dividere gli organismi in parti differenti selezionando ciascun tessuto potrebbe anche fornire l'indicazione se l'animale produce da sé il principio attivo o se esso proviene dalla dieta o ancora è associato a parassiti o microrganismi. La maggior parte degli AMP marini sono stati di fatto isolati dal compartimento sangue, sia dall’emolinfa/fluido celomatico (sangue intero) o dagli emociti/celomociti (cellule del sangue). Alcuni peptidi tuttavia sono stati scoperti e isolati da altri tessuti, ma essendo questi circondati (negli invertebrati marini) dagli emociti/celomociti, è possibile che provengano da questi ultimi. Inoltre tutte le variabili analitiche hanno una potenziale influenza sul risultato sia in termini quantitativi che qualitativi oltre che per la stabilità del campione anche per la riproducibilità del protocollo. Tra queste variabili ricadono le condizioni di conservazione del campione (tempo trascorso dall’estrazione, la temperatura prima dell’estrazione ed alla separazione, il congelamento e la liofilizzazione), la tipologia del campione ed il metodo di prelievo, la separazione delle cellule dal sangue (velocità di centrifugazione, durata e temperatura), l’uso del tampone (tipo, pH, forza ionica e temperatura), l'uso e il tipo di anticoagulanti e cocktail inibitori della proteasi. Inoltre, dati bibliografici riportano come la stimolazione immunitaria determini un aumento sia quantitativamente che qualitativamente della produzione di peptidi antimicrobici. Per tale motivo si è proceduto ad effettuare un esperimento pilota per valutare se in tali condizioni fosse superiore la quantità di piccoli peptidi sintetizzati rispetto ad individui non stimolati e se nei differenti tempi di stimolazione selezionati, emergesse una differenziazione a carico dei peptidi prodotti in relazione ad una possibile attività di natura antibatterica

Characterization of translationally controlled tumour protein from the sea anemone Anemonia viridis and transcriptome wide identification of cnidarian homologues

Gene family encoding translationally controlled tumour protein (TCTP) is defined as highly conserved among organisms; however, there is limited knowledge of non-bilateria. In this study, the first TCTP homologue from anthozoan was characterised in the Mediterranean Sea anemone, Anemonia viridis. The release of the genome sequence of Acropora digitifera, Exaiptasia pallida, Nematostella vectensis and Hydra vulgaris enabled a comprehensive study of the molecular evolution of TCTP family among cnidarians. A comparison among TCTP members from Cnidaria and Bilateria showed conserved intron exon organization, evolutionary conserved TCTP signatures and 3D protein structure. The pattern of mRNA expression profile was also defined in A. viridis. These analyses revealed a constitutive mRNA expression especially in tissues with active proliferation. Additionally, the transcriptional profile of A. viridis TCTP (AvTCTP) after challenges with different abiotic/biotic stresses showed induction by extreme temperatures, heavy metals exposure and immune stimulation. These results suggest the involvement of AvTCTP in the sea anemone defensome taking part in environmental stress and immune responses

Gene expression changes after parental exposure to metals in the sea urchin affect timing of genetic programme of embryo development

It is widely accepted that phenotypic traits can be modulated at the epigenetic level so that some conditions can affect the progeny of exposed individuals. To assess if the exposure of adult animals could result in effects on the offspring, the Mediterranean sea urchin and its wellcharacterized gene regulatory networks (GRNs) was chosen as a model. Adult animals were exposed to known concentrations of zinc and cadmium (both individually and in combination) for 10 days, and the resulting embryos were followed during the development. The oxidative stress occurring in parental gonads, embryo phenotypes and mortality, and the expression level of a set of selected genes, including members of the skeletogenic and endodermal GRNs, were evaluated. Increased oxidative stress at F0, high rates of developmental aberration with impaired gastrulation, in association to deregulation of genes involved in skeletogenesis (dri, hex, sm50, p16, p19, msp130), endodermal specification (foxa, hox11/13b, wnt8) and epigenetic regulation (kat2A, hdac1, ehmt2, phf8 and UBE2a) occurred either at 24 or 48 hpf. Results strongly indicate that exposure to environmental pollutants can affect not only directly challenged animals but also their progeny (at least F1), influencing optimal timing of genetic programme of embryo development, resulting in an overall impairment of developmental success

Procedures for high quality RNA extraction from Paracentrotus lividus (LAMARCK, 1816) embryos and gonadal tissue

The extraction protocol described below is part of the CISAS project, aimed at the realization, within CNR, of an “International Centre of advanced study in environment, ecosystem and human health”. In this report we describe a RNA extraction protocol from Paracentrotus lividus embryos and gonadal tissue