Amyloid fibrils and amorphous aggregates are two types of aberrant aggregates associated with protein misfolding diseases. Although they differ in morphology, the two forms are often treated indiscriminately. β 2 -microglobulin (β2m), a protein responsible for dialysis-related amyloidosis, forms amyloid fibrils or amorphous aggregates depending on the NaCl concentration at pH 2.5. We compared the kinetics of their formation, which was monitored by measuring thioflavin T fluorescence, light scattering, and 8-anilino-1-naphthalenesulfonate fluorescence. Thioflavin T fluorescence specifically monitors amyloid fibrillation, whereas light scattering and 8-anilino-1-naphthalenesulfonate fluorescence monitor both amyloid fibrillation and amorphous aggregation. The amyloid fibrils formed via a nucleation-dependent mechanism in a supersaturated solution, analogous to crystallization. The lag phase of fibrillation was reduced upon agitation with stirring or ultrasonic irradiation, and disappeared by seeding with preformed fibrils. In contrast, the glass-like amorphous aggregates formed rapidly without a lag phase. Neither agitation nor seeding accelerated the amorphous aggregation. Thus, by monitoring the kinetics, we can distinguish between crystal-like amyloid fibrils and glass-like amorphous aggregates. Solubility and supersaturation will be key factors for further understanding the aberrant aggregation of proteins.protein aggregation | metastability | glass transition | ultrasonication

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This study is devoted to deducing exact elastic constants of an anisotropic solid material without using any advance information on the elastic constants by incorporating a displacement-distribution measurement into resonant ultrasound spectroscopy (RUS). The usual RUS method measures free-vibration resonance frequencies of a solid and compares them with calculations to find the most suitable set of elastic constants by an inverse calculation. This comparison requires mode identification for the measured resonance frequencies, which has been difficult and never been free from ambiguity. This study then adopts a laser-Doppler interferometer to measure the displacement-distribution patterns on a surface of the vibrating specimen mounted on pinducers; comparison of the measured displacement distributions with those computed permits us to correctly identify the measured resonance frequencies, leading to unmistakable determination of elastic constants. Because the displacement patterns are hardly affected by the elastic constants, an exact answer is surely obtained even when unreasonable elastic constants are used as initial guesses at the beginning of the inverse calculation. The usefulness of the present technique is demonstrated with an aluminum alloy and a langasite crystal.

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