PEMF and red blood cells

Magnawave on Rouleau (stacks of red blood cells) and Bionuu Clinic have YouTube videos claiming that PEMF makes red blood cells (RBC) less sticky. A Russian language peer reviewed PEMF publications on the PEMF exposure of RBC ghosts warrants further investigation as it involves a protein sysem important in redox balance. Why RBC appear to be sticky on slides in the first place could signal a potential therapy PEMF. We have so much more to learn about how PEMF affects RBC

A reason why RBCs aggregate on clinical slides

In 2004 Shlomo Berliner and Israeli coauthors published a study comparing RBC aggregation in peripheral blood glass slides (PBGS) and a cell flow analyzer (CFA). Of importance to practitioners, a reduction in RBC aggregation following LDL apheresis , a method for removing low density lipoprotein particles from whole blood,was noted in PBGS and CFA. The following is a direct quote from the text simply because Ca²⁺ figures so predominantly in PEMF literature. RBC aggregation does not have to reach the threshold of Rouleau to b bad.

It should be emphasized that different calcium
chelating agents are used in the two different systems herewith described (sodium citrate for the slide test and EDTA for the computerized flow analyzer). Moreover, the two systems are
operated with different hematocrits. In the slide test, there is a 25% dilution with the anticoagulant (1 vol. of citrate with 3 vol. of whole blood) while in the flow chamber, a hematocrit of 6% is reached following the suspension of the washed erythrocytes in the autologous plasma. Despite these differences in anticoagulants and hematocrits, the significant correlation found between the systems is suggestive for a final common biological principle, namely the inter-relations. . (skipping text). . . .We conclude that the phenomenon of RBC aggregation as determined by our slide test is governed to a significant degree by inter-erythrocytic cohesive forces and is not a result of a chancesuper imposition of the cells on each other. This demonstrates that the slide method can be employed for a facile assessment of RBC aggregation and can be used for routine test of abnormal RBC aggregation in pathological conditions.

We can speculate that un-disclosed details in PEMF sessions that appear to decrease RBC aggregation on PBGS might be sloughing off LDL particles and/or Ca²⁺. For those that like images of LDL cholesterol gluing RBC together, this is a LINK to one of dozens on the Internet.

very low frequency PEMF and RBC “ghosts”

Loginov 1991 was published in Russian with an English abstract. Just the abstract can lead us to other clinically relevant insights into PEMF and RBC. Red blood cell “ghosts” were prepared by hypotonic lysis. Ideally these ghosts have all of the surface proteins but none of the innards like hemoglobin. Smaller stuff was separated from larger stuff using a Millipore filter. Owing to charges on the surface, these ghosts move in an electric field in a process called electrophoresis. The matrix was a highly porous mixture of polyacrylamide and agarose.

After exposure to 1-10 Hz pulses at a field intensity of 24 mT it was determined that there was a decrease in the amount of mount actin and glyceraldehyde-phosphate dehydrogenase(GAPDH). The effects of PEMF are explained by the changes in the distribution of electrostatic charges near the membrane protein-protein binding sites.

GAPDH binds to RBC band 3

Biochemists know GAPDH as an abundant glycolytic enzyme involved in glucose metabolism. Never mind the historical reason for calling the chloride bicarbonate exchngerin RBC membranes “band 3.” The anion exchanger is a newer alias. IGAPDH seems to associate with Band 3 . In 1980 KIinman and Steck studied the kinetics of GAPDH from Band 3 in response to EDTA, NaCl, reducing equivalent NADH, and changes in the dielectric constant of the solution by adding methanol.

Bronsted-Bjerrum analysis suggested that two or three charges of opposite sign on each protein were involved in binding. This would be a “secondary” effect in which a salt alters the concentration of a reactant by binding to it. The primary effect would be activity coefficient and hence the rate of reaction. In the Kliman and Steck 1980 NADH appeared to elute the enzyme by interaction with its catalytic site, but the band 3 binding site extended beyond the NADH site, permitting the formation of a ternary NADH- enzyme-membrane complex. GAPDH is a branch point to the pentose phosphate pathway (PPP) that generates the reducing equivalent NADPH important in defense against oxidative stress.

What could all of this mean for banked blood?

Lorentzian forces generated by PEMF de-aggregating RBC could be a good thing that deserved more rigorous verification, particularly if LDL particles are the glue (Berliner 2004). PEMF induced release of GAPDH from RBC membranes containing Band 3 (Loginov 1991) could be a not so good thing given the NADH controlled interaction between the two (Kliman & Streck 1986) Issaian 2021 wrote a very comprehensive review of the redox regulation of the Band 3 (anion exchanger AE1)/ GAPDH complex and other binding partners that include hemoglobin. High levels of oxidative stress favor the binding of Band 3/AE1 to GAPDH inhibiting enzyme activity. This favors the NADPH generating PPP. A quote from the conclusion:

Through a combination of proteomics approaches (co-immunoprecipitation, thermal proteome profiling and crosslinking proteomics) we validated previously described interactors (e.g., GAPDH) and identified potential new candidate binding partners for AE11-56. Some of the interactions reported here may also be nonspecific (e.g., those driven by electrostatic interactions with the negatively charged acidic residue-rich N-terminus of AE1) or affected by environmental conditions (e.g., low vs. high oxygen saturation). Further studies will address whether a membrane-penetrating version of the AE1 N-terminus peptide could rescue oxidant stress-induced lesions to RBC AE1 in the context of those pathologies that have been reported to target the N-terminus of AE1, including COVID-19.

We have so much to learn about how PEMF combinations affect red blood cells. How important is frequency, magnetic field intensity and the wave shape? Can PEMF positively benefit blood in the patient as well as blood going into storage as well as coming out of storage?

References

• Berliner S, Ben-Ami R, Samocha-Bonet D, Abu-Abeid S, Schechner V, Beigel Y, Shapira I, Yedgar S, Barshtein G. The degree of red blood cell aggregation on peripheral blood glass slides corresponds to inter-erythrocyte cohesive forces in laminar flow. Thromb Res. 2004;114(1):37-44. Sci-Hub free paper
• Kliman HJ, Steck TL. Association of glyceraldehyde-3-phosphate dehydrogenase with the human red cell membrane. A kinetic analysis. J Biol Chem. 1980 Jul 10;255(13):6314-21. free paper
• Loginov VA. Izmenenie zariada éritrotsitarnoĭ membrany pri bozdeĭstvii impul’snogo élektromagnitnogo polia [Change in charge of the erythrocyte membrane during exposure to a pulsed electromagnetic field]. Biofizika. 1991 Jul-Aug;36(4):614-20. Russian. PMID: 1793745.
• Issaian A, Hay A, Dzieciatkowska M, Roberti D, Perrotta S, Darula Z, Redzic J, Busch MP, Page GP, Rogers SC, Doctor A, Hansen KC, Eisenmesser EZ, Zimring JC, D’Alessandro A. The interactome of the N-terminus of band 3 regulates red blood cell metabolism and storage quality. Haematologica. 2021 Nov 1;106(11):2971-2985. PMC free paper.