Covalent Immobilization of Aldehyde and Alcohol Dehydrogenases on Ordered Mesoporous Silicas

Purpose This work studies the immobilization of two enzymes, the alcohol dehydrogenase (ADH) and the aldehyde dehydrogenase (AldDH) both from Saccharomyces cerevisiae, which could be used to produce high value-added molecules from carboxylic acids embedded in anaerobic digestate.Methods In particular, three mesoporous siliceous materials, with different specific surface areas and pore sizes, (MSU-H, MSU-F and MCF0.75) were used as supports for covalent immobilization. The support materials were characterized by complementary techniques. Then, after a functionalization, creating a covalent bond between the enzyme and the support was performed. The specific activity and immobilization yield of the biocatalysts were then evaluated.Results The best results were obtained with MSU-H and MSU-F, resulting in an immobilization yield greater than 50% in all cases, a specific activity of 0.13 IU/g(supp) with the AldDH/MSU-H, 0.10 IU/g(supp) with AldDH/MSU-F, 48.6 IU/g(supp) with ADH/MSU-H and 12.6 IU/g(supp) with ADH/MSU-H. These biocatalysts were then characterized by optimal pH and temperature and the stability factor was evaluated. With ADH/MSU-F no decrease in activity was observed after 120 h incubated at 50 degrees C. Finally, the biocatalysts AldDH/MSU-H and ADH/MSU-H were used to perform the reduction reaction and it was seen that after five reaction cycles the residual activity was greater than 20% in both cases.Conclusion The ADH and AldDH enzymes have been successfully immobilized on mesoporous siliceous supports, considerably increasing their thermal stability and being able to reuse them for several reaction cycles. The use of this immobilization and these supports is adaptable to a wide variety of enzymes

Tramesan elicits durum wheat defense against the septoria disease complex

The Septoria Leaf Blotch Complex (SLBC), caused by the two ascomycetes Zymoseptoria tritici and Parastagonospora nodorum, can reduce wheat global yearly yield by up to 50%. In the last decade, SLBC incidence has increased in Italy; notably, durum wheat has proven to be more susceptible than common wheat. Field fungicide treatment can efficiently control these pathogens, but it leads to the emergence of resistant strains and adversely affects human and animal health and the environment. Our previous studies indicated that active compounds produced by Trametes versicolor can restrict the growth of mycotoxigenic fungi and the biosynthesis of their secondary metabolites (e.g., mycotoxins). Specifically, we identified Tramesan: a 23 kDa α-heteropolysaccharide secreted by T. versicolor that acts as a pro-antioxidant molecule in animal cells, fungi, and plants. Foliar-spray of Tramesan (3.3 μM) on SLBC-susceptible durum wheat cultivars, before inoculation of causal agents of Stagonospora Nodorum Blotch (SNB) and Septoria Tritici Blotch (STB), significantly decreased disease incidence both in controlled conditions (SNB: –99%, STB: –75%) and field assays (SNB: –25%, STB: –30%). We conducted these tests were conducted under controlled conditions as well as in field. We showed that Tramesan increased the levels of jasmonic acid (JA), a plant defense-related hormone. Tramesan also increased the early expression (24 hours after inoculation-hai) of plant defense genes such as PR4 for SNB infected plants, and RBOH, PR1, and PR9 for STB infected plants. These results suggest that Tramesan protects wheat by eliciting plant defenses, since it has no direct fungicidal activity. In field experiments, the yield of durum wheat plants treated with Tramesan was similar to that of healthy untreated plots. These results encourage the use of Tramesan to protect durum wheat against SLBC

Neuroimmagini e profilo neuropsicologico nell’adrenoleucodistrofia legata all’X

Background scientifico: L'adrenoleucodistrofia a trasmissione recessiva legata all'X (ALD-X) è una patologia neurodegenerativa severa associata ad una progressiva demielinizzazione del sistema nervoso centrale e periferico, insufficienza surrenale (malattia di Addison) ed un accumulo di acidi grassi a catena molto lunga (VLCFA) nel plasma, nei fibroblasti e nei tessuti. Nell'8% dei casi origina da nuove mutazioni. Le forme parieto-occipitali sono le più frequenti. Scopo: I primi sintomi consistono in difficoltà cognitive, a livello visuomotorio e visuospaziale. Il Nostro scopo è quello di valutare se vi sia una discrepanza tra neuroimmagini e profilo neuropsicologico nell'ALD-X. Metodologie e soggetti: Riportiamo il caso di un bambino di 8,6 anni che giunge al pronto soccorso per un episodio simil-comiziale insorto in apiressia, al risveglio, con perdita di urine e seguito da ipotonia generalizzata, rallentamento nei movimenti e nell'eloquio ed amnesia retrograda. Effettuate EEGvv, TAC e RMN encefalo si sospetta ALD. Il bambino viene inviato presso il Reparto Degenze del Nostro Dipartimento. In anamnesi presente astenia e problemi uditivi da circa sei mesi per cui ha effettuato un videat ORL risultato nella norma. La gravidanza è normodecorsa, con sviluppo psicomotorio normale. Genitori non consanguinei entrambi italiani. Il bambino durante il ricovero è stata effettuata: misurazione degli VLCFA e dell'ACTH nel plasma, cortisolemia, esame del campo visivo centrale cromatico, EEGvv, RMN encefalo in spettroscopia, PEV, SEP, BAER, Sensory NCS, Motor NCS, F Wave. Lui, la madre ed il fratello minore effettuano l'analisi molecolare del gene ABCD1. Inoltre, il bambino è stato sottoposto ad una valutazione neuropsicologica. Una volta confermata la diagnosi e valutata l'estensione della lesione che non permetteva di effettuare il trapianto di midollo osseo, il bambino inizia terapia sostitutiva con idrocortisone ed Olio di Lorenzo; da circa sei mesi è entrato a far parte di un protocollo sperimentale per una nuova terapia. Attualmente abbiamo un follow-up di due anni. Risultati e discussioni: Si rilevano un aumento dei valori degli VCLFA e dell'ACTH. Il bambino presenta una mutazione puntiforme de novo per il gene ABCD1 (c.346G>A); la madre ed il fratello sono sani. All'EEGvv si evidenziano anomalie lente posteriori. Risultano nella norma: cortisolemia, PEV, SEP, BAER, Sensory NCS, Motor NCS, F Wave. All'esame del campo visivo centrale cromatico si evidenzia un restringimento irregolare della soglia differenziale visiva per tutti gli stimoli bilateralmente, prevalente per quello rosso. Alla RMN si rileva una diffusa, simmetrica e marcata demielinizzazione predominante a livello parietale. Alla spettroscopia si evidenzia un'elevazione del picco di colina con riduzione del picco del NAA e comparsa del picco del Lattato. L'esame trattografico dimostra rarefazione delle fibre mieliniche nella sostanza bianca occipito-parietale bilateralmente. Tutte queste alterazioni permangono stabili nel tempo. Dalla Nostra valutazione sono state registrate oscillazioni nel profilo neuropsicologico del bambino nelle varie età, con andamento non omogeneo, soprattutto in quei test che valutano le capacità motorie e di integrazione visuomotorie e nei test che valutano gli apprendimenti scolastici (soprattutto della letto-scrittura e della matematica). Queste oscillazioni, non coincidono con il quadro neuroradiologico rimasto invariato nei due anni di follow-up dimostrando così una discrepanza esistente tra neuroimmagini e profilo neuropsicologico. Tutto ciò ci porta a mettere in evidenza quanto sia importante, nei paziente affetti da ALD, integrare le indagini neuroradiologiche con una approfondita valutazione anche a livello neuropsicologico

Covalent Immobilization of Dehydrogenases on Carbon Felt for Reusable Anodes with Effective Electrochemical Cofactor Regeneration

This study presents the immobilization with aldehyde groups (glyoxyl carbon felt) of alcohol dehydrogenase (ADH) and formate dehydrogenase (FDH) on carbon-felt-based electrodes. The compatibility of the immobilization method with the electrochemical application was studied with the ADH bioelectrode. The electrochemical regeneration process of nicotinamide adenine dinucleotide in its oxidized form (NAD(+)), on a carbon felt surface, has been deeply studied with tests performed at different electrical potentials. By applying a potential of 0.4 V versus Ag/AgCl electrode, a good compromise between NAD(+) regeneration and energy consumption was observed. The effectiveness of the regeneration of NAD(+) was confirmed by electrochemical oxidation of ethanol catalyzed by ADH in the presence of NADH, which is the no active form of the cofactor for this reaction. Good reusability was observed by using ADH immobilized on glyoxyl functionalized carbon felt with a residual activity higher than 60 % after 3 batches

Aflatoxin Control in Maize by Trametes versicolor

Aspergillus flavus is a well-known ubiquitous fungus able to contaminate both in pre- and postharvest period different feed and food commodities. During their growth, these fungi can synthesise aflatoxins, secondary metabolites highly hazardous for animal and human health. The requirement of products with low impact on the environment and on human health, able to control aflatoxin production, has increased. In this work the effect of the basidiomycete Trametes versicolor on the aflatoxin production by A. flavus both in vitro and in maize, was investigated. The goal was to propose an environmental loyal tool for a significant control of aflatoxin production, in order to obtain feedstuffs and feed with a high standard of quality and safety to enhance the wellbeing of dairy cows. The presence of T. versicolor, grown on sugar beet pulp, inhibited the production of aflatoxin B1 in maize by A. flavus. Furthermore, treatment of contaminated maize with culture filtrates of T. versicolor containing ligninolytic enzymes, showed a significant reduction of the content of aflatoxin B1

Preoperative chemoradiotherapy affects postoperative outcomes and functional results in patients treated with transanal endoscopic microsurgery for rectal neoplasms

Background: The aim of this study was to quantify the incidence of short-term postoperative complications and functional disorders at 1 year from transanal endoscopic microsurgery (TEM) for rectal neoplasms, to compare patients treated with TEM alone and with TEM after preoperative chemoradiotherapy (CRT) and to analyse factors influencing postoperative morbidity and functional outcomes. Methods: A retrospective study was conducted on all patients treated with TEM for rectal neoplasms at our institution in January 2000–December 2017. Data from a prospectively maintained database were retrospectively analysed. Patients were divided into two groups: adenoma or early rectal cancer (no CRT group) and locally advanced extraperitoneal rectal cancer with major or complete clinical response after preoperative CRT (CRT group). Short-term postoperative mortality and morbidity and the functional results at 1 year were recorded. The two groups were compared, and a statistical analysis of factors influencing postoperative morbidity and functional outcomes was performed. Functional outcome was also evaluated with the low anterior resection syndrome (LARS) score (0–20 no LARS, 21–29 minor LARS and 30–42 major LARS). Results: One hundred and thirteen patients (71 males, 42 females, median age 64 years [range 41–80 years]) were included in the study (46 in the CRT group). The overall postoperative complication rate was 23.0%, lower in the noCRT group (p < 0.001), but only 2.7% were grade ≥ 3. The most frequent complication was suture dehiscence (17.6%), which occurred less frequently in the noCRT group (p < 0.001). At 1 year from TEM, the most frequent symptoms was urgency (11.9%, without significant differences between the CRT group and the noCRT group); the noCRT group experienced a lower rate of soiling than the CRT group (0% vs. 7.7%; p: 0.027). The incidence of LARS was evaluated in 47 patients from May 2012 on and was 21.3% occurring less frequently in the noCRT group (10% vs. 41.2%; p: 0.012). Only 6.4% of the patients evaluated experienced major LARS. In multivariate analysis, preoperative CRT significantly worsened postoperative morbidity and functional outcomes. Conclusions: TEM is a safe procedure associated with only low risk of severe postoperative complications and major LARS. Preoperative CRT seems to increase the rate of postoperative morbidity after TEM and led to worse functional outcomes at 1 year after surgery

Augmented reality (AR) in minimally invasive surgery (MIS) training: where are we now in Italy? The Italian Society of Endoscopic Surgery (SICE) ARMIS survey

Minimally invasive surgery (MIS) is a widespread approach in general surgery. Computer guiding software, such as the augmented reality (AR), the virtual reality (VR) and mixed reality (MR), has been proposed to help surgeons during MIS. This study aims to report these technologies' current knowledge and diffusion during surgical training in Italy. A web-based survey was developed under the aegis of the Italian Society of Endoscopic Surgery (SICE). Two hundred and seventeen medical doctors’ answers were analyzed. Participants were surgeons (138, 63.6%) and residents in surgery (79, 36.4%). The mean knowledge of the role of the VR, AR and MR in surgery was 4.9 ± 2.4 (range 1–10). Most of the participants (122, 56.2%) did not have experience with any proposed technologies. However, although the lack of experience in this field, the answers about the functioning of the technologies were correct in most cases. Most of the participants answered that VR, AR and MR should be used more frequently for the teaching and training and during the clinical activity (170, 80.3%) and that such technologies would make a significant contribution, especially in training (183, 84.3%) and didactic (156, 71.9%). Finally, the main limitations to the diffusion of these technologies were the insufficient knowledge (182, 83.9%) and costs (175, 80.6%). Based on the present study, in Italy, the knowledge and dissemination of these technologies are still limited. Further studies are required to establish the usefulness of AR, VR and MR in surgical training