Preparation of an engineered safer immunotoxin against colon carcinoma based on the ribotoxin hirsutellin A

Immunotoxins are chimeric proteins composed of an antibody domain that specifically directs the action of the toxic domain, resulting in the death of the targeted cells. Over recent years, immunotoxins have been widely studied and the number of different constructions has increased exponentially. Protein engineering has allowed the design of optimized versions of immunotoxins with an improved tumor binding affinity, stability or cytotoxic efficacy, although sometimes this has compromised the safety of the patient in terms of undesirable adverse secondary reactions. A triple mutant at three Trp residues (HtA3DW) of the ribotoxin hirsutellin A retains its specific ribonucleolytic activity, although cell internalization capacity is lacking.This toxin variant has been fused to the single chain variable fragment A33 (scFvA33). This immunoconjugate (IMTXA33HtA3DW) was produced in the methylotrophic yeast Pichia pastoris and purified using nickelnitrilotriacetic acid affinity chromatography. Both target and toxic domains were characterized. The immunotoxin showed an exquisite specific binding against GPA33-positive culture cells, which results in the death of the targeted cells because of specific ribonucleolytic activity against ribosomes of the engineered hirsutellin A variant. IMTXA33HtA3DW represents a promising structure in the search for an improved immunotoxin without compromising the safety of patients.

The left visual field attentional advantage: No evidence of different speeds of processing across visual hemifields.

Temporal-order judgment (TOJ) and simultaneity judgment (SJ) tasks are used to study differences in speed of processing across sensory modalities, stimulus types, or experimental conditions. Matthews and Welch (2015) reported that observed performance in SJ and TOJ tasks is superior when visual stimuli are presented in the left visual field (LVF) compared to the right visual field (RVF), revealing an LVF advantage presumably reflecting attentional influences. Because observed performance reflects the interplay of perceptual and decisional processes involved in carrying out the tasks, analyses that separate out these influences are needed to determine the origin of the LVF advantage. We re-analyzed the data of Matthews and Welch (2015) using a model of performance in SJ and TOJ tasks that separates out these influences. Parameter estimates capturing the operation of perceptual processes did not differ between hemifields by these analyses, whereas parameter estimates capturing the operation of decisional processes differed. In line with other evidence, perceptual processing also did not differ between SJ and TOJ tasks. Thus, the LVF advantage occurs with identical speeds of processing in both visual hemifields. If attention is responsible for the LVF advantage, it does not exert its influence via prior entry.