High predictive values of RBC membrane-based diagnostics by biophotonics in an integrated approach for Autism Spectrum Disorders

Abstract

Membranes attract attention in medicine, concerning lipidome composition and fatty acid correlation with neurological diseases. Hyperspectral dark field microscopy (HDFM), a biophotonic imaging using reflectance spectra, provides accurate characterization of healthy adult RBC identifying a library of 8 spectral end-members. Here we report hyperspectral RBC imaging in children affected by Autism Spectrum Disorder (ASD) (n = 21) compared to healthy age-matched subjects (n = 20), investigating if statistically significant differences in their HDFM spectra exist, that can comprehensively map a membrane impairment involved in disease. A significant difference concerning one end-member (spectrum 4) was found (P value = 0.0021). A thorough statistical treatment evidenced: i) diagnostic performance by the receiving operators curve (ROC) analysis, with cut-offs and very high predictive values (P value = 0.0008) of spectrum 4 for identifying disease; ii) significant correlations of spectrum 4 with clinical parameters and with the RBC membrane deficit of the omega-3 docosahexaenoic acid (DHA) in ASD patients; iii) by principal component analysis, very high affinity values of spectrum 4 to the factor that combines behavioural parameters and the variable “cc” discriminating cases and controls. These results foresee the use of biophotonic methodologies in ASD diagnostic panels combining with molecular elements for a correct neuronal growth.

The Erythrocyte Membrane Lipidome of Healthy Dogs: Creating a Benchmark of Fatty Acid Distribution and Interval Values

“…The carrier gas was hydrogen, held at a constant pressure of 16.482 psi. FAMEs were identified by comparison with the retention times of standard references (see Supplementary Figure 2 for a representative chromatogram) in agreement with previously reported procedures, satisfactorily separating all the 10 fatty acids, without superimposition of other peaks (15,16,26,29). The group of 10 fatty acids corresponds to chromatographic peak areas >97%, and consisted of: two SFAs (palmitic and stearic acids); three MUFAs (palmitoleic, oleic, and cis-vaccenic acids); three PUFA omega-6 (linoleic, dihomogamma linolenic, arachidonic acids); and two PUFA omega-3 (eicosapentaenoic and docosahexaenoic acids).…”

“…In human studies, we contributed to research in membrane fatty-acid-based lipidomics, making the arbitrary choice of analyzing a specific fatty acid cohort of erythrocyte membrane glycerophospholipids, made of 10 fatty acids representative of SFA, MUFA, and PUFA families. Their interval values in healthy cohorts were reported in membrane fatty acids by several studies, also by us (25)(26)(27), and in one of the most complete metaanalysis appeared in the literature (28). Taking into account that novel fatty acid pathways and transformations are continuously studied (29), it is important to make the strategic choice of a specific number of fatty acids considering their fundamental biochemical/biological roles and the consensus reached about their levels in cell membranes.…”

Na+, K+‐ATPase activity in children with autism spectrum disorder: Searching for the reason(s) of its decrease in blood cells

“…Noteworthy, Cornelius et al stressed that, because cholesterol does not mix well with DHA, the effect of the two lipids on the conformation of NKA is the opposite; cholesterol increases the lateral pressure in the middle of the bilayer and decreases it near the interfaces, whereas DHA has the opposite effects. Because erythrocyte membranes from ASD patients show depleted DHA [Ghezzo et al, ; Giacometti et al, ], the effect of a relative depletion in both cholesterol and DHA could have on NKA activity may be worth examining in a future work. Likewise, contrary to what we have observed, in kidney cells a decrease in membrane cholesterol was reported to cause a reduction in α1 expression and viceversa [Chen et al, ].…”

Fatty Acids and Membrane Lipidomics in Oncology: A Cross-Road of Nutritional, Signaling and Metabolic Pathways

“…Based on these considerations, the fatty acid profile can give information on the balance of these molecular components in erythrocyte phospholipids and help to establish the changes occurring under healthy and unhealthy conditions. Indeed, the fatty acid-based membrane lipidome monitoring used in different human conditions revealed how the endogenously and exogenously-derived fatty acids of erythrocytes are affected [ 61 , 62 , 63 , 64 ]. In Table 2 , the relevant data relating to erythrocyte fatty acid monitoring from studies on cancer patients are gathered, highlighting the cancer types, the country and the number of patients and detailing the most important conclusions of each study.…”