METHODS TO DETERMINE COMPLEX DEGREES OF MUTUAL ANISOTROPY FOR THE DIFFERENTIATION OF HEMORRHAGES IN THE HUMAN BRAIN SUBSTANCE RESULTING FROM TRAUMATIC AND NON-TRAUMATIC GENESIS

An issue that is often debated in forensic traumatology is the differential diagnosis of hemorrhages into the human brain substance (HBS) of traumatic and non-traumatic genesis. Objectives. This study aims to identify new criteria for objective forensic differentiation of hemorrhages of traumatic origin, cerebral infarction of ischemic and hemorrhagic genesis by using the method of complex degree of mutual anisotropy. For this study native sections of HBS from 125 corpses were used in the case of: death from coronary heart disease - 35 (28%) of native sections (Group 1 - control); hemorrhages of traumatic genesis - 30 (24%) sections (Group 2); ischemic cerebral infarction - 30 (24%) native sections(Group 3); and hemorrhages of non-traumatic genesis - 30 (24%) native sections (Group 4). Results. The statistical moments of the third and fourth orders, which characterize the asymmetry and excess of the complex degree of mutual anisotropy module size distributions, the strength of the method of polarization-correlation microscopy in the differentiation of the samples of the histological sections of the brain of control and experimental groups reached a good level — 87%-90%. Conclusion. The method of complex degree of mutual anisotropy allows differentiating with great precision the genesis of hemorrhage into the substance of the brain.

Cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post-translational modification

The effects of the immunophilins, FKBP12 and FKBP12.6, and phosphorylation on type II ryanodine receptor (RyR2) arrangement and function were examined using correlation microscopy (line scan confocal imaging of Ca2+ sparks and dual-tilt electron tomography) and dSTORM imaging of permeabilized Wistar rat ventricular myocytes. Saturating concentrations (10 µmol/L) of either FKBP12 or 12.6 significantly reduced the frequency, spread, amplitude and Ca2+ spark mass relative to control, while the tomograms revealed both proteins shifted the tetramers into a largely side-by-side configuration. Phosphorylation of immunophilin-saturated RyR2 resulted in structural and functional changes largely comparable to phosphorylation alone. dSTORM images of myocyte surfaces demonstrated that both FKBP12 and 12.6 significantly reduced RyR2 cluster sizes, while phosphorylation, even of immunophilin-saturated RyR2, increased them. We conclude that both RyR2 cluster size and the arrangement of tetramers within clusters is dynamic and respond to changes in the cellular environment. Further, these changes affect Ca2+ spark formation.