Quantum based effects of therapeutic nuclear magnetic resonance persistently reduce glycolysis

Therapeutic nuclear magnetic resonance and intermittent hypoxia trigger time dependent on/off effects in circadian clocks and confirm a central role of superoxide in cellular magnetic field effects

“…NIH3T3 cells stably expressing a Per2:Luc construct [ 13 ] were cultured and exposed to sham or tNMR in either 21% or 1% of oxygen as described [ 18 ]. Cells were cultured under constant darkness, only interrupted by short dim light periods during the processes of seeding and medium change (see Supplemental Figure 1 ).…”

“…Primers for Cry1 and Cry2 mRNA were already used in Ref. [ 18 ]. Primers for Bmal1 mRNA (Accession number NM_007489.4) were designed using Primer Express 3.0 (Applied Biosystems, USA), optimizing the primer concentrations via primer matrices prior to the real-time qPCR; Bmal1 f orward primer, 5′-GAC GAA GAC AAT GAG CCA GAC A-3′ and reverse primer, 5′-CGC CCG ATT GCA ACG A-3’.…”

“…Western blots and normalization to amounts of total protein were conducted as described [ 18 ], see Supplemental Figure 1, B ). As primary antibodies mouse monoclonal CRY1 (Santa Cruz Biotechnology, sc393466, 1:500), CRY2 (Abgent AM8637b, also known as Q49AN0, 1:4000) and BMAL1 (Cell signaling technology, D2L7G, 1:2000) were used.…”

“…Measurement of mitochondrial H 2 O 2 was performed as described [ 18 ]. Briefly, the ro-GFP2-Orp1 expressing cells were seeded at a density of 15,000 cells per well and the medium was changed the following day.…”

“…Notably, we recently showed that tNMR without any additional light source increased the mitochondrial and extracellular pools of H 2 O 2 and redirected hypoxia induced glycolysis towards mitochondrial respiration [ 18 ]. Our results align with similar effects observed under RF fields of 1.4 MHz, 50 μT static field, and a field intensity of 10 μT [ 19 ].…”