1. Towards a New Unified Theory ofDisease The Clinical Significanceof the Jones-Ray Effect

2. Robert M. Davidson1 and Stephanie Seneff 21InternalMedicine Group Practice, PhyNet, Inc.Longview, TX 75605, USA; E-Mail:patrons99@yahoo.com2ComputerScience and Artificial IntelligenceLaboratory, MIT, 32 Vassar Street, Cambridge, MA01890, USA; E-Mail:seneff@csail.mit.edu 2 3. Outline Zeta Potential The Riddick Effect The Jones-Ray Effect The Hofmeister Effect Coherence Domains Interfacial Tension and Engulfment Free Energy of Engulfment Interfacial Water Stress (IWS) Wave Phenomena, Energy and Water Stressors Quantum-coherent Bio-interfacial Waves Electron Micrographs Illustrating Phenomena Clinically Important Observations Summary and Take-away Message3 4. Zeta Potential, definedthe electrical potential drop from the particlesurface across the bound fluid, to the interfacewhere the liquid begins to flow under theshear stress.the zeta potential is the potential at thesurface boundary between the stationary fluidand the liquid that is moving with theparticle.- Tigrek and Barnes (2010) 4 5. The Riddick EffectZeta potential data for various electrolytes in an anionic colloidal suspension of 100ppm Minusil . Data were originally published per Thomas M. Riddick (1968) and isreproduced with permission of Zeta-Meter, Inc. 5 6. The Jones-Ray EffectThe points are the original data from 1937 by Jones and Ray [154]. The change in thesurface tension of all 13 Jones-Ray salts were fitted to a simple model by Petersen andSaykally (2005). Reproduced here with permission from Journal of the American ChemicalSociety.6 7. Fluctuations and the Hofmeister Effect:A Brief History Hofmeister (1888) ordered anions according to their ability to precipitateglobular proteins from water. Setschenow (1889) established an empirical law linking the solubility of aprotein with cosolute (salt) concentration . Heydweiller (1910) discovered that salt dissolved in water increased the surfacetension of the solution-air interface . Langmuir (1917) was the first to attempt a theoretical explanation of thephysical mechanism behind the increase in surface tension produced byelectrolytes. Der et al. (2007, 2008), instead of focusing on air-water surface tension, usedprotein-water interfacial tension as a general description of the free energychanges associated with salt-induced changes of protein solubility andconformation .7 8. The Fluctuation-Dissipation Theorem First proven by Callen and Welton in 1951. - Describes how dissipative forces and fluctuatingrandom forces are connected. Fluctuations in protein conformation are linked tointerfacial tension and protein structural stability. Many proteins oscillate between open and closedconformations - Implies water-exposed surface area changes. A general relationship exists between salt concentrationand protein-water interfacial tension. - Plays a key role in protein structure and dynamics. - Unfolded protein response is a common stressorleading to cellular apoptosis. 8 9. QED Coherence in MatterGiuliano Preparatas (1942-2000) QED Coherence in Matter [1995]provides the basis for understanding the Stable, Non-EquilibriumSystem we call Life 9 10. Embryological Envelopment is Biophysically-drivenDAH Steinbergs Differential Adhesion HypothesisDITH Brodlands Differential Interfacial Tension HypothesisR. A. Foty, C. M. Pfleger, G. Forgacs, andM. S. Steinberg. Surface tensions ofembryonic tissues predict their mutualenvelopment behavior.Development, 122(5):161120, 1996. 0950-1991Schoetz, E., Dynamics and Mechanics ofZebrafish Embryonic Tissues. Der Fakultatfur Physik der Technischen UniversitatDresden, 2007.Brodland, G. W., The Differential InterfacialTension Hypothesis (DITH): a comprehensivetheory for the self-rearrangement of embryoniccells and tissues. Journal of biomechanicalengineering 2002, 124 (2), 188-97.10 11. Moore, P. L.; Bank, H. L.; Brissie, N. T.;Spicer, S. S., Phagocytosis of bacteria bypolymorphonuclear leukocytes. A freeze-fracture, scanning electronmicroscope, and thin-sectioninvestigation of membrane structure.The Journal of cell biology 1978, 76(1), 158-74. 11 12. Highly-StereotypedBiophysically-driven Processes Engulfment Envelopment Endocytosis Exocytosis Macropinocytosis Vesiculation Podokinesis Transcellular Diapedesis Immune Cell Activation Fusion Adhesion Nutritive Endocytosis Autophagy 12 13. Free Energy of Engulfment where PB is the phagocyte-bacterium interfacial tension and BL the bacterium-liquid interfacial tension. An empirical Equation of State was developed for determining interfacial tensions. used in conjunction with Youngs equationto obtain the interfacial tension between liquid and solid bymeasuring the contact angle and surface tension of the liquid.Absolom, Darryl R, Measurement of surface properties of phagocytes, bacteria, and other particles. In Methods inEnzymology, Giovanni Di Sabato, J. E., Ed. Academic Press: 1986; Vol. Volume 132, pp 16-95.13 14. Contact Angle and Surface Tension Measurement yields the Free Energy of Engulfment14 15. Absolom, DarrylR, Measurement ofsurface properties ofphagocytes, bacteria, and other particles. InMethods inEnzymology, GiovanniDi Sabato, J. E., Ed.Academic Press: 1986;Vol. Volume 132, pp 16-95. 15 16. Interfacial Water StressWe refer to water stress as a propertyof interfacial water - interfacialtension - which destabilizesenzymes, protein structure, and cellmembranes 16 17. Intravital MicroscopyW T Coakley, et al. have detected wavephenomena at the biointerfaces of RBCs,platelets, and endothelial cells.The glycocalyx layers surrounding our cellmembranes have been shown by intravitalmicroscopists to have properties similar tothe massive EZs discovered by GeraldPollack.17 18. 18 19. Average number of waves perwavy cell rimdecreased when cell surface charge wasdepleted, when polyvalent cations were in thesuspending phase, and when cationic drugswere present, and increased in the presence ofanionic drugs.Gallez, D.; Coakley, W. T., Interfacial instability at cellmembranes. Progress in Biophysics and Molecular Biology1986, 48 (3), 155-199.19 20. Wave Phenomena, Energy and Water Stressors Wave phenomena have been detected at the biointerfacesof RBCs, platelets, and endothelial cells- Longer wavelengths in the membrane imply lower energy- Lower energy implies energy-unloading of the membrane- Higher surface tension implies intramembrane instability, longerwavelengths, and lower energy in the membrane.- Lower surface tension implies intramembrane stability, shorterwavelengths, and greater energy in the membrane. Exogenous interfacial water "stressors" in the lowmicromolar concentration range energy-unload our cellmembranes. Examples of exogenous interfacial water stressors:- Polycationic surfactants (strong kosmotropic cations)- Certain non-ionic surfactants (especially in the low pH range) 20 21. Wave Phenomena at Bio-interfacesBiophysical Manifestations of Cellular Stress Quantum coherent bio-interfacial water is resonating in-phase over all timescales. The Wave Equation applies. 21 22. The Wave Equation e = c x h/where is wavelength, h is Plancksconstant, and c is the speed oflight, indicates that the longer thewavelength, the lower the energy. 22 23. Hypothesis Endothelial Glycocalyx Layers (EGL)described by intravital microscopistsrepresent Exclusion Zones (EZs) analogousto those described by Gerald Pollack. Interfacial water found in membrane-bound caveolae of lipid rafts provides aquantum coherent ensemble of CoherenceDomains (CDs) analogous to thosedescribed by Emilio Del Giudice.23 24. HypothesisA modified dissipative version of the waveequation applies to: wave phenomena in cell membranes waves in EZs waves in a quantum coherent ensemble ofCDs.Waves in the RBC membrane are coherentwith waves in the EZ and CD around the RBCmembrane. 24 25. Quantum coherent Bio-interfacial WavesResonating in phase, even though thetime scales may be vastly differentbetween the membrane, EZ, and CDwaves. 25 26. An Important Role for Cholesterol Sulfate Cholesterol Sulfate (Ch-S) reduces interfacial instability at cell membranes by reducing Exogenous Interfacial Water Stress (EIWS). When the intramembrane wavelength is longer, the intramembrane energy is lower, and the exogenous interfacial water stressors are energy-unloading the membrane, i.e. energy-unloading the exclusions zones (EZs) and coherence domains (CDs) of our cell membranes.26 27. Cholesterol Sulfate (Ch-S) reducesinterfacial instability at cell membranes by reducing ExogenousInterfacial Water Stress (EIWS).Hydrophobic coreKosmotropic anion27 28. 28 29. (a) Scanning electron micrograph of human erythrocytes inhypotonic saline solution. X 10,000.(b) Scanning electron micrograph of human erythrocytes inhypotonic saline solution. X 20,000.(c) Scanning electron micrograph of human erythrocytes inhypotonic saline solution in presence of 10-5 Mcholesterol sulfate. X 10,000.Bleau, G.; Lalumiure, G.; Chapdelaine, A.; Roberts, K., Redcell surface structure. Stabilization by cholesterol sulfate asevidenced by scanning electron microscopy. Biochimica etbiophysica acta 1975, 375 (2), 220-3. 29 30. 30 31. 31 32. 32 33. Brecher, G.; Bessis, M., Present status of spiculed red cells and their relationship to the discocyte-echinocyte transformation:a critical review. Blood 1972, 40 (3), 333-44. 33 34. 34 35. 35 36. 36 37. SEM image of a red blood cell and a white blood cell stacked on top of the red blood cell.Adapted from John F. Lesoine, Electron Microscopy of Human Blood Cellshttp://www.optics.rochester.edu/workgroups/cml/opt307/spr06/john/lesoineTEMSEM.hm 37 38. An SEM image showing two different stages of a platelet becoming activated.Adapted from John F. Lesoine, Electron Microscopy of Human Blood Cellshttp://www.optics.rochester.edu/workgroups/cml/opt307/spr06/john/lesoineTEMSEM.htm 38 39. Adapted from John F. Lesoine, Electron Microscopy of Human Blood Cellshttp://www.optics.rochester.edu/workgroups/cml/opt307/spr06/john/lesoineTEMSEM.htm 39 40. Some Clinically-important ObservationsPredicted by the Jones-Ray Effect Molecular Mimicry of Cholesterol Sulfate Membrane stabilization by Cholesterol Sulfate of RBCmembranes subjected to hypotonic stress Effect of free fatty acids on Erythrocytes - Erythrocytes protected against hypotonic hemolysis in low concentration range, but opposite effect in high concentration - Biphasic free fatty acid chain length dependence observed in human erythrocytes. Sulfated neurosteroids are potent non-competitiveantagonists of GABAA receptors without a clear structureactivity relationship. Zeta potential measurements of blood samples may provide asurrogate marker for interfacial tensions40 41. Summary and Takeaway Message The Jones-Ray Effect is not accidental nor is it just a curiosity. The in vivo concentration of sulfate in arterial blood occurs at athermodynamic minimum in both the surface enhancement and the zetapotential enhancement. The biphasic shape of the Jones-Ray and Zeta potential curves hasimportant implications for the role of the biosulfates in modulatinginterfacial tension, membrane potential, and zeta potential of our blood. As a surrogate marker of interfacial tension, the biphasic Jones-Ray Effectprovides a plausible explanation as to why hydrophobic anions potently anduncompetitively antagonize receptor function in the absence of aconventional binding site.41 42. 42