Over the last 50 years, the incidence of human thyroid cancer disease has seen a significative increment. This comes along with an even higher increment of surgery, since, according to the international guidelines, patients are sometimes addressed to surgery also when the fine needle aspiration gives undetermined cytological diagnosis. As a matter of fact, only 30% of the thyroid glands removed for diagnostic purpose have a post surgical histological report of malignancy: this implies that about 70% of the patients have suffered an unnecessary thyroid removal. Here we show that Raman spectroscopy investigation of thyroid tissues provides reliable cancer diagnosis. Healthy tissues are consistently distinguished from cancerous ones with an accuracy of ∼ 90%, and the three cancer typology with highest incidence are clearly identified. More importantly, Raman investigation has evidenced alterations suggesting an early stage of transition of adenoma tissues into cancerous ones. These results suggest that Raman spectroscopy may overcome the limits of current diagnostic tools.

Ultrastructural fluctuations at nanoscale are fundamental to assess properties and functionalities of advanced out-of-equilibrium materials. We have taken myelin as a model of supramolecular assembly in out-of-equilibrium living matter. Myelin sheath is a simple stable multilamellar structure of high relevance and impact in biomedicine. Although it is known that myelin has a quasi-crystalline ultrastructure, there is no information on its fluctuations at nanoscale in different states due to limitations of the available standard techniques. To overcome these limitations, we have used scanning micro X-ray diffraction, which is a unique non-invasive probe of both reciprocal and real space to visualize statistical fluctuations of myelin order of the sciatic nerve of Xenopus laevis. The results show that the ultrastructure period of the myelin is stabilized by large anticorrelated fluctuations at nanoscale, between hydrophobic and hydrophilic layers. The ratio between the total thickness of hydrophilic and hydrophobic layers defines the conformational parameter, which describes the different states of myelin. Our key result is that myelin in its out-of-equilibrium functional state fluctuates point-to-point between different conformations showing a correlated disorder described by a Levy distribution. As the system approaches the thermodynamic equilibrium in an aged state, the disorder loses its correlation degree and the structural fluctuation distribution changes to Gaussian. In a denatured state at low pH, it changes to a completely disordered stage. Our results aim to clarify the degradation mechanism in biological systems by associating these states with ultrastructural dynamic fluctuations at nanoscale.

While the ultrastructure of myelin is considered a quasi-crystalline stable system, nowadays its multiscale complex dynamics appear to play a key role in its functionality, degeneration and repair processes following neurological diseases and trauma. In this work, we investigated the fluctuation of the myelin supramolecular assembly by measuring the spatial distribution of orientation fluctuations of axons in a Xenopus Laevis sciatic nerve associated with nerve functionality. To this end, we used scanning micro X-ray diffraction (SµXRD), a non-invasive technique that has already been applied to other heterogeneous systems presenting complex geometries from microscale to nanoscale. We found that the orientation of the spatial fluctuations of fresh axons show a Levy flight distribution, which is a clear indication of correlated disorder. We found that the Levy flight distribution was missing in the aged nerve prepared in an unfresh state. This result shows that the spatial distribution of axon orientation fluctuations in unfresh nerve state loses the correlated disorder and assumes a random disorder behavior. This work provides a deeper understanding of the ultrastructure-function nerve relation and paves the way for the study of other materials and biomaterials using the SµXRD technique to detect fluctuations in their supramolecular structure.