Different routes to pure food allergens: extraction, recombinant techniques and total chemical synthesis for obtaining the plant-food pan-allergen LTP

La richiesta di allergeni puri è in continuo aumento per scopi diagnostici, come standard per metodi di rilevamento e di quantificazione, per l'immunoterapia e per lo studio a livello molecolare dei meccanismi delle reazioni allergiche, al fine di facilitare lo sviluppo di possibili cure. In questa tesi di dottorato sono descritte diverse strategie per l’ottenimento di forme pure di non-specific Lipid Transfer Proteins (nsLTPs), le quali sono state riconosciute essere rilevanti allergeni alimentari in molti frutti e verdure comunemente consumati e sono state definite come modello di veri allergeni alimentari. Una LTP potenzialmente allergenica, non nota in precedenza, è stata isolata dalle mandorle, mentre una LTP dall’allergenicità nota contenuta nelle noci è stata prodotta mediante tecniche di DNA ricombinante. Oltre a questi approcci classici, metodi per la sintesi chimica totale di proteine sono stati applicati per la prima volta alla produzione di un allergene, utilizzando Pru p 3, la LTP prototipica e principale allergene della pesca nell'area mediterranea, come modello. La sintesi chimica totale di proteinepermette di controllarne completamente la sequenza e di studiare la loro funzione a livello atomico. La sua applicazione alla produzione di allergeni costituisce perciò un importante passo avanti nel campo della ricerca sulle allergie alimentari. La proteina Pru p 3 è stata prodotta nella sua intera lunghezza e sono necessari solo due passaggi finali di deprotezione per ottenere il target nella sua forma nativa. Le condizioni sperimentali per tali deprotezioni sono state messe a punto durante la produzione dei peptidi sPru p 3 (1-37) e sPru p 3 (38-91), componenti insieme l'intera proteina. Tecniche avanzate di spettrometria di massa sono state usate per caratterizzare tutti i composti ottenuti, mentre la loro allergenicità è stata studiata attraverso test immunologici o approcci in silico.

UV femtosecond laser inscribes a 300 nm period nanostructure in a pure fused silica

We report on the first recording of a periodic structure of ∼150 nm pitch in a permanently moving sample of a pure fused silica using the tightly focused, 82 nJ, 267 nm, 300 fs, 1 kHz laser pulses. © 2007 IOP Publishing Ltd.

Are faces special? A case of pure prosopagnosia

The ability to recognize individual faces is of crucial social importance for humans and evolutionarily necessary for survival. Consequently, faces may be “special” stimuli, for which we have developed unique modular perceptual and recognition processes. Some of the strongest evidence for face processing being modular comes from cases of prosopagnosia, where patients are unable to recognize faces whilst retaining the ability to recognize other objects. Here we present the case of an acquired prosopagnosic whose poor recognition was linked to a perceptual impairment in face processing. Despite this, she had intact object recognition, even at a subordinate level. She also showed a normal ability to learn and to generalize learning of nonfacial exemplars differing in the nature and arrangement of their parts, along with impaired learning and generalization of facial exemplars. The case provides evidence for modular perceptual processes for faces.

Effects of the build-up and resetting of auditory stream segregation on temporal discrimination

The tendency to hear a tone sequence as 2 or more streams (segregated) builds up, but a sudden change in properties can reset the percept to 1 stream (integrated). This effect has not hitherto been explored using an objective measure of streaming. Stimuli comprised a 2.0-s fixed-frequency inducer followed by a 0.6-s test sequence of alternating pure tones (3 low [L]-high [H] cycles). Listeners compared intervals for which the test sequence was either isochronous or the H tones were slightly delayed. Resetting of segregation should make identifying the anisochronous interval easier. The HL frequency separation was varied (0-12 semitones), and properties of the inducer and test sequence were set to the same or different values. Inducer properties manipulated were frequency, number of onsets (several short bursts vs. one continuous tone), tone:silence ratio (short vs. extended bursts), level, and lateralization. All differences between the inducer and the L tones reduced temporal discrimination thresholds toward those for the no-inducer case, including properties shown previously not to affect segregation greatly. Overall, it is concluded that abrupt changes in a sequence cause resetting and improve subsequent temporal discrimination. (PsycINFO Database Record © 2009 APA, all rights reserved)

Auditory stream segregation of tone sequences in cochlear implant listeners

Previous claims that auditory stream segregation occurs in cochlear implant listeners are based on limited evidence. In experiment 1, eight listeners heard tones presented in a 30-s repeating ABA-sequence, with frequencies matching the centre frequencies of the implant's 22 electrodes. Tone A always stimulated electrode 11 (centre of the array); tone B stimulated one of the others. Tone repetition times (TRTs) from 50 to 200 ms were used. Listeners reported when they heard one or two streams. The proportion of time that each sequence was reported as segregated was consistently greater with increased electrode separation. However, TRT had no significant effect, and the perceptual reversals typical of normal-hearing listeners rarely occurred. The results may reflect channel discrimination rather than stream segregation. In experiment 2, six listeners performed a pitch-ranking task using tone pairs (reference = electrode 11). Listeners reported which tone was higher in pitch (or brighter in timbre) and their confidence in the pitch judgement. Similarities were observed in the individual pattern of results for reported segregation and pitch discrimination. Many implant listeners may show little or no sign of automatic stream segregation owing to the reduced perceptual space within which sounds can differ from one another. © 2006 Elsevier B.V. All rights reserved.

Femtosecond inscription of the first order Bragg gratings in pure fused silica

The fabrication of sub-micron periodic structures beyond diffraction limit is a major motivation for the present paper. We describe the fabrication of the periodic structure of 25 mm long with a pitch size of 260 nm which is less than a third of the wavelength used. This is the smallest reported period of the periodic structure inscribed by direct point-by-point method. A prototype of the add-drop filter, which utilizes such gratings, was demonstrated in one stage fabrication process of femtosecond inscription, in the bulk fused silica.

Femtosecond inscription of the first order Bragg gratings in pure fused silica

The fabrication of sub-micron periodic structures beyond diffraction limit is a major motivation for the present paper. We describe the fabrication of the periodic structure of 25 mm long with a pitch size of 260 nm which is less than a third of the wavelength used. This is the smallest reported period of the periodic structure inscribed by direct point-by-point method. A prototype of the add-drop filter, which utilizes such gratings, was demonstrated in one stage fabrication process of femtosecond inscription, in the bulk fused silica.

Inscription of a 300-nm-period nanostructure in a pure fused silica

Very recently, using tightly-focused femtosecond near-IR pulses, periodical sub-micron structures have been recorded [1,2]. Such microfabrication utilizes the multi-photon approach, which allows the inscription inside various non-photosensitive optical materials. The combination of multi-photon excitation with the point-by-point technique offers the great potential of creating non-uniform chirped gratings by controlling the rate of femtosecond pulses or the sample translation speed.

UV femtosecond laser inscribes a 300 nm period nanostructure in a pure fused silica

We report on the first recording of a periodic structure of ~150 nm pitch in a permanently moving sample of a pure fused silica using the tightly focused, 82 nJ, 267 nm, 300 fs, 1 kHz laser pulses.

Simultaneous grouping in cochlear implant listeners:can abrupt changes in level be used to segregate components from a complex tone?

A sudden increase in the amplitude of a component often causes its segregation from a complex tone, and shorter rise times enhance this effect. We explored whether this also occurs in implant listeners (n?=?8). Condition 1 used a 3.5-s “complex tone” comprising concurrent stimulation on five electrodes distributed across the array of the Nucleus CI24 implant. For each listener, the baseline stimulus level on each electrode was set at 50% of the dynamic range (DR). Two 1-s increments of 12.5%, 25%, or 50% DR were introduced in succession on adjacent electrodes within the “inner” three of those activated. Both increments had rise and fall times of 30 and 970 ms or vice versa. Listeners reported which increment was higher in pitch. Some listeners performed above chance for all increment sizes, but only for 50% increments did all listeners perform above chance. No significant effect of rise time was found. Condition 2 replaced amplitude increments with decrements. Only three listeners performed above chance even for 50% decrements. One exceptional listener performed well for 50% decrements with fall and rise times of 970 and 30 ms but around chance for fall and rise times of 30 and 970 ms, indicating successful discrimination based on a sudden rise back to baseline stimulation. Overall, the results suggest that implant listeners can use amplitude changes against a constant background to pick out components from a complex, but generally these must be large compared with those required in normal hearing. For increments, performance depended mainly on above-baseline stimulation of the target electrodes, not rise time. With one exception, performance for decrements was typically very poor.