Processi di biorefining per l'estrazione di secondary chemical building blocks da sottoprotti dell'agro-industria

Phenol and cresols represent a good example of primary chemical building blocks of which 2.8 million tons are currently produced in Europe each year. Currently, these primary phenolic building blocks are produced by refining processes from fossil hydrocarbons: 5% of the world-wide production comes from coal (which contains 0.2% of phenols) through the distillation of the tar residue after the production of coke, while 95% of current world production of phenol is produced by the distillation and cracking of crude oil. In nature phenolic compounds are present in terrestrial higher plants and ferns in several different chemical structures while they are essentially absent in lower organisms and in animals. Biomass (which contain 3-8% of phenols) represents a substantial source of secondary chemical building blocks presently underexploited. These phenolic derivatives are currently used in tens thousand of tons to produce high cost products such as food additives and flavours (i.e. vanillin), fine chemicals (i.e. non-steroidal anti-inflammatory drugs such as ibuprofen or flurbiprofen) and polymers (i.e. poly p-vinylphenol, a photosensitive polymer for electronic and optoelectronic applications). European agrifood waste represents a low cost abundant raw material (250 millions tons per year) which does not subtract land use and processing resources from necessary sustainable food production. The class of phenolic compounds is essentially constituted by simple phenols, phenolic acids, hydroxycinnamic acid derivatives, flavonoids and lignans. As in the case of coke production, the removal of the phenolic contents from biomass upgrades also the residual biomass. Focusing on the phenolic component of agrifood wastes, huge processing and marketing opportunities open since phenols are used as chemical intermediates for a large number of applications, ranging from pharmaceuticals, agricultural chemicals, food ingredients etc. Following this approach we developed a biorefining process to recover the phenolic fraction of wheat bran based on enzymatic commercial biocatalysts in completely water based process, and polymeric resins with the aim of substituting secondary chemical building blocks with the same compounds naturally present in biomass. We characterized several industrial enzymatic product for their ability to hydrolize the different molecular features that are present in wheat bran cell walls structures, focusing on the hydrolysis of polysaccharidic chains and phenolics cross links. This industrial biocatalysts were tested on wheat bran and the optimized process allowed to liquefy up to the 60 % of the treated matter. The enzymatic treatment was also able to solubilise up to the 30 % of the alkali extractable ferulic acid. An extraction process of the phenolic fraction of the hydrolyzed wheat bran based on an adsorbtion/desorption process on styrene-polyvinyl benzene weak cation-exchange resin Amberlite IRA 95 was developed. The efficiency of the resin was tested on different model system containing ferulic acid and the adsorption and desorption working parameters optimized for the crude enzymatic hydrolyzed wheat bran. The extraction process developed had an overall yield of the 82% and allowed to obtain concentrated extracts containing up to 3000 ppm of ferulic acid. The crude enzymatic hydrolyzed wheat bran and the concentrated extract were finally used as substrate in a bioconversion process of ferulic acid into vanillin through resting cells fermentation. The bioconversion process had a yields in vanillin of 60-70% within 5-6 hours of fermentation. Our findings are the first step on the way to demonstrating the economical feasibility for the recovery of biophenols from agrifood wastes through a whole crop approach in a sustainable biorefining process.

"Crop elicitation": approccio innovativo per la valorizzazione delle proprietà funzionali in leguminose da granella

Crop elicitation: innovative approach for the valorization of grain legume functional properties. In Italy grain legume cultivation has encountered a drastic decrease due to several causes (productive, economic, social). In this regard, studies aimed at the setting up of agronomic techniques able to guarantee high and constant in planta yields of health-promoting compounds may concur at re-launching legume production. In this context, 22 accessions of grain legumes (17 Phaseolus vulgaris, 3 Phaseolus coccineus, 1 Vigna unguiculata and 1 Glycine max genotypes) were screened with the aim of identifying genotypes rich in health beneficial phytochemicals (α-amylase inhibitors, α -glucosidase inhibitors, polyphenols) and with low anti-nutritional compounds (lectins). A wide variability was observed among investigated accessions. Four genotypes (Verdone, Kidney Cina, Roviotto and DG) showed a α -amylase inhibitory activity significantly higher (approximately 30% more) than all other tested accessions. The α -amylase inhibitory activity was not correlated neither with the protein nor with the polyphenol contents. Conversely, the α -glucosidase inhibitory activity was positively correlated with grain color and polyphenol content: dark-colored seeds had a mean inhibitory activity of 83.64 ± 22.07%, whereas light-colored seeds had mean values of 21.11 ± 9.36%. As regards the anti-nutritional compounds, out of all common bean accessions, only DG showed no erythro-agglutination activity (lectins). Preliminary experiments, performed in controlled environment, permitted to highlight that different germination conditions markedly affect the synthesis and accumulation of functional compounds in legume seedlings. Those findings were confirmed with field trials performed in two different locations (Bologna and Pisa), on two bean genotypes (Verdone and Zolfino), during the 2004-2005 cropping season. Results showed that the application of abiotic stresses (no fertilization and /or no irrigation) lead to a significant increase of flavonoids in grains, but a decrease (up to 50%) in legume yields was also observed. Crop elicitation, even if valuable for boosting health-promoting compound synthesis in crops, must necessary cope with economically acceptable crop yields.

Fusarium species responsible for mycotoxin production in wheat crop: involvement in food safety

Fusarium Head Blight (FHB) is a worldwide cereal disease responsible of significant yield reduction, inferior grain quality, and mycotoxin accumulation. Fusarium graminearum and F. culmorum are the prevalent causal agents. FHB has been endemic in Italy since 1995, while there are no records about its presence in Syria. Forty-eight and forty-six wheat kernel samples were collected from different localities and analyzed for fungal presence and mycotoxin contamination. Fusarium strains were identified morphologically but the molecular confirmation was performed only for some species. Further differentiation of the chemotypes for trichothecene synthesis by F. graminearum and F. culmorum strains was conducted by PCR assays. Fusarium spp. were present in 62.5% of Syrian samples. 3Acetyl-Deoxynivalenol and nivalenol chemotypes were found in F. culmorum whilst all F. graminearum strains belonged to NIV chemotype. Italian samples were infected with Fusarium spp for 67.4%. 15Ac-DON was the prevalent chemotype in F. graminearum, while 3Ac-DON chemotype was detected in F. culmorum. The 60 Syrian Fusarium strains tested for mycotoxin production by HPLC-MS/MS have shown the prevalence of zearalenone while the emerging mycotoxins were almost absent. The analysis of the different Syrian and Italian samples of wheat kernels for their mycotoxin content showed that Syrian kernels were mainly contaminated with storage mycotoxins, aflatoxins and ochratoxin whilst Italian grains with mainly Fusarium mycotoxins. The aggressiveness of several Syrian F. culmorum isolates was estimated using three different assays: floret inoculation in growth chamber, ear inoculation in the field and a validated new Petri-dish test. The study of the behaviour of different Syrian wheat cultivars, grown under different conditions, has revealed that Jory is a FHB Syrian tolerant cultivar. This is the first study in Syria on Fusarium spp. associated to FHB, Fusarium mycotoxin producers and grain quality.

Valutazione dell'effetto del mezzo temprante sulle tensioni residue e proprieta meccaniche della lega di alluminio Al-Si7-Mg

L’alluminio, grazie alla sua bassa temperatura di fusione e all’elevata fluidità di molte leghe, è uno dei metalli più versatili in fonderia. Per ottenere il massimo delle proprietà resistenziali, le leghe che lo permettono sono sottoposte a trattamento termico. Questo, però, in componenti geometricamente complessi può far insorgere tensioni residue che inficeranno la resistenza del materiale durante la sua messa in esercizio. Gli obiettivi della presente ricerca sono stati: la valutazione dell’incidenza del mezzo temprante usato durante il trattamento termico sulle tensioni residue che si sviluppano nella lega AlSi7Mg e la definizione di come questo potesse essere modificato per ridurre le tensioni senza peggiorare le proprietà resistenziali del materiale. Per tale ragione sono state eseguite prove di trattamento termico, analisi microstrutturali e test di durezza su campioni estratti direttamente da getti colati in sabbia.

Determinazione delle tensioni residue in componenti metallici sottili mediante "incremental hole drilling"

La vita di un aeromobile è un elemento importante sia per quanto riguarda la sicurezza del payload, che da un punto di vista economico, a causa delle spese di manutenzione/rinnovamento della flotta che una compagnia aerea deve affrontare. Gli elementi costitutivi di un aeromobile sono soggetti a diverse tipologie di carichi, alcuni dei quali ciclici come la pressurizzazione/depressurizzazione della fusoliera; tali carichi, nel lungo periodo, possono provocare la nascita e la propagazione di eventuali cricche, le quali possono portare alla rottura del componente stesso causando gravi incidenti. Il legame tra tensioni residue e nascita/crescita delle cricche ha portato allo sviluppo di tecniche per contrastare questo fenomeno, come il processo del LSP. Per la misura delle tensioni residue esistono già normative di riferimento, le quali però non trattano componenti metallici di piccolo spessore mentre i pannelli di fusoliera rientrano in questa categoria. Scopo di questa tesi è quello di studiare una variante della tecnica HD adatta a valutare le tensioni residue in componenti metallici di piccolo spessore e confrontare i risultati con quelli ottenuti con la tecnica XRD. L’idea di partenza è l’implementazione di un supporto posteriore in resina che simuli la presenza di uno spessore maggiore.

A residue close to ?1 loop F disrupts modulation of GABAA receptors by benzodiazepines while their binding is maintained

Benzodiazepines act at the major isoforms of GABA type A receptors where they potentiate the current evoked by the agonist GABA. The underlying mechanism of this potentiation is poorly understood, but hypothesized to be related to the mechanism that links agonist binding to channel opening in these ligand activated ion channels. The loop F of the ?(1) and the ?(2) subunit have been implicated in channel gating, and loop F of the ?(2) subunit in the modulation by benzodiazepines. We have identified the conservative point mutation Y168F located N-terminally of loop F in the ?(1) subunit that fails to affect agonist properties. Interestingly, it disrupts modulation by benzodiazepines, but leaves high affinity binding to the benzodiazepine binding site intact. Modulation by barbiturates and neurosteroids is also unaffected. Residue ?(1) Y168 is not located either near the binding pockets for GABA, or for benzodiazepines, or close to the loop F of the ?(2) subunit. Our results support the fact, that broader regions of ligand gated receptors are conformationally affected by the binding of benzodiazepines. We infer that also broader regions could contribute to signaling from GABA agonist binding to channel opening.

Mutagenesis and computer modeling studies of a GPCR conserved residue W5.43(194) in ligand recognition and signal transduction for CB2 receptor

W5.43(194), a conserved tryptophan residue among G-protein coupled receptors (GPCRs) and cannabinoid receptors (CB), was examined in the present report for its significance in CB2 receptor ligand binding and adenylyl cyclase (AC) activity. Computer modeling postulates that this site in CB2 may be involved in the affinity of WIN55212-2 and SR144528 through aromatic contacts. In the present study, we reported that a CB2 receptor mutant, W5.43(194)Y, which had a tyrosine (Y) substitution for tryptophan (W), retained the binding affinity for CB agonist CP55940, but reduced binding affinity for CB2 agonist WIN55212-2 and inverse agonist SR144528 by 8-fold and 5-fold, respectively; the CB2 W5.43(194)F and W5.43(194)A mutations significantly affect the binding activities of CP55940, WIN55212-2 and SR144528. Furthermore, we found that agonist-mediated inhibition of the forskolin-induced cAMP production was dramatically diminished in the CB2 mutant W5.43(194)Y, whereas W5.43(194)F and W5.43(194)A mutants resulted in complete elimination of downstream signaling, suggesting that W5.43(194) was essential for the full activation of CB2. These results indicate that both aromatic interaction and hydrogen bonding are involved in ligand binding for the residue W5.43(194), and the mutations of this tryptophan site may affect the conformation of the ligand binding pocket and therefore control the active conformation of the wild type CB2 receptor. W5.43(194)Y/F/A mutations also displayed noticeable enhancement of the constitutive activation probably attributed to the receptor conformational changes resulted from the mutations.

Evaluation of the chemical residue monitoring in animal-derived products in Switzerland

This paper evaluates whether the Swiss monitoring programme for foreign substances in animal products fulfils basic epidemiological quality requirements, and identifies possible sources of bias in the selection of samples. The sampling was analysed over a 4-year period (2002-05). The sampling frame in 37 participating abattoirs covered 51% of all slaughtered pigs, 73% of calves, 68% of beef and 36% of cows. The analysis revealed that some sub-populations as defined by the region of origin were statistically over-represented while others were under-represented. The programme that is in accordance with European Union requirements contained some relevant bias. Patterns of under-sampled regions characterized by management type differences were identified. This could lead to an underestimate of the number of contaminated animals within the programme. Although the current sampling was stratified and partially risk-based, its efficiency could be improved by adopting a more targeted approach.

Planting and production of switchgrass (Panicum virgatum L.) as a bioenergy crop in Michigan's Upper Peninsula

Switchgrass (Panicum virgatum L.) is a perennial grass holding great promise as a biofuel resource. While Michigan’s Upper Peninsula has an appropriate land base and climatic conditions, there is little research exploring the possibilities of switchgrass production. The overall objectives of this research were to investigate switchgrass establishment in the northern edge of its distribution through: investigating the effects of competition on the germination and establishment of switchgrass through the developmental and competitive characteristics of Cave-in-Rock switchgrass and large crabgrass (Digitaria sanguinalis L.) in Michigan’s Upper Peninsula; and, determining the optimum planting depths and timing for switchgrass in Michigan’s Upper Peninsula. For the competition study, a randomized complete block design was installed June 2009 at two locations in Michigan’s Upper Peninsula. Four treatments (0, 1, 4, and 8 plants/m2) of crabgrass were planted with one switchgrass plant. There was a significant difference between switchgrass biomass produced in year one, as a function of crabgrass weed pressure. There was no significant difference between the switchgrass biomass produced in year two versus previous crabgrass weed pressure. There is a significant difference between switchgrass biomass produced in year one and two. For the depth and timing study, a completely randomized design was installed at two locations in Michigan’s Upper Peninsula on seven planting dates (three fall 2009, and four spring 2010); 25 seeds were planted 2 cm apart along 0.5 m rows at depths of: 0.6 cm, 1.3 cm, and 1.9 cm. Emergence and biomass yields were compared by planting date, and depths. A greenhouse seeding experiment was established using the same planting depths and parameters as the field study. The number of seedlings was tallied daily for 30 days. There was a significant difference in survivorship between the fall and spring planting dates, with the spring being more successful. Of the four spring planting dates, there was a significant difference between May and June in emergence and biomass yield. June planting dates had the most percent emergence and total survivorship. There is no significant difference between planting switchgrass at depths of 0.6 cm, 1.3 cm, and 1.9 cm. In conclusion, switchgrass showed no signs of a legacy effect of competition from year one, on biomass production. Overall, an antagonistic effect on switchgrass biomass yield during the establishment period has been observed as a result of increasing competing weed pressure. When planting switchgrass in Michigan’s Upper Peninsula, it should be done in the spring, within the first two weeks of June, at any depth ranging from 0.6 cm to 1.9 cm.