Biophysical Aspects of the Sanarelli-Shwartzman Phenomenon

Robert M. Davidson M.D. Ph.D. FAIS*Ann Lauritzen, Stephanie Seneff,

Stephen D. Kette, Glyn Wainwright, Anthony Samsel, and Sydney J. Bush

*Fellow, The American Institute of StressPhysician and Medical Researcher, Kilgore, Texas, USA

Email: patrons99@yahoo.com

3rd International Conference on Hematology & Blood Disorders November 2-4, 2015 Atlanta, Georgia USA

1

DEFINITION:

UNIVERSAL NON-SPECIFIC MESENCHYMAL REACTION (UNMR)

• The UNMR refers to the different causes of an acceleration of metabolism of connective tissue of blood vessels as well as that of all organs studied [Hauss et al. 1962].

• Selye (1966) described a local or general intravascular

coagulation that occurs in response to certain toxins or systemic stress [Selye, Thrombohemorrhagic Phenomena, 1966].

2

DEFINITION:

Sanarelli-Shwartzman Phenomenon (SSP), a.k.a. Generalized Shwartzman Reaction (GSR)

• Activation of the coagulation system is thought to be the primary event in a clinical syndrome involving disseminated intravascular coagulation (DIC), a consumptive coagulopathy of the microvasculature, which can be localized or generalized, acute, subacute, or chronic.

• Fibrinolysis, if it occurs, is thought to be a secondary protective phenomenon [Bang et al. Thrombosis and Bleeding Disorders, 2014]

3

The Major “Players" in Our Passion Play

Acid mucopolysaccharides

• Serine proteases

• Aryl sulfatases

• Transition metals

• Inflammatory stress

• Interfacial water stress

4

TIME IS OF THE ESSENCE

A prompt response is often required: there’s not much time for de novo protein synthesis and diffusion of substrates

• In the setting of acute bleeding: to achieve hemostasis and maintain organ perfusion

• In the setting of critical care medicine: associated with inflammatory stress, e.g., ARDS, DIC syndrome, severe trauma, acute coronary and neurovascular syndromes

• In the setting severe infectious disease, e.g. septicemia, Purpura fulminans, Hemorrhagic Fever viruses (Dengue, Ebola, etc.)

5

6

Purpura Fulminans

“a rare syndrome of intravascular thrombosis and hemorrhagic infarction of the skin that is rapidly progressive and is accompanied by vascular collapse and disseminated intravascular coagulation. It may be classified as (1) neonatal, (2) idiopathic, or (3) acute infectious.” The 4 primary features of acute infectious purpura fulminans are as follows:

• Large purpuric skin lesions • Fever • Hypotension • Disseminated intravascular coagulation (DIC)

SOURCE: http://emedicine.medscape.com/article/2202749-overview

THE INITIAL COMMON PATHWAY

Exogenous Interfacial Water Stress (EIWS)

Inflammation

Disease

7

Davidson, Robert M.; Seneff, Stephanie. 2012. Entropy 14, no. 8: 1399-1442.

Davidson, R.M.; Lauritzen, A.; Seneff, S. 2013. Entropy 15, 3822-3876.

Two Important Characteristics of Inflammatory States

Low pH: 4.7 - 5.7 Tsai, et al (2014)

• cardiac ischemia • cancer • septicemia• trauma (fractures, hematomas)

High oxidative stress: Khansari et al (2009); Reuter et al (2010); Kim et al (2013)

8

The Redox Potential of Water is pH Dependent

“The redox potential increases as the pH decreases (increasing [H+])”. --- Martin Chaplin

9http://www1.lsbu.ac.uk/water/water_redox.html

Our Proposal for pH Dependent e(aq) Speciation

10

Mizuse, K. and Fujii, A. (2013). Characterization of a Solvent-Separated Ion-Radical Pair in Cationized Water Networks: Infrared Photodissociation and Ar-Attachment Experiments for Water Cluster Radical Cations (H2O)n+ (n = 3–8). The Journal of Physical Chemistry A, 117, 929-938.Larsen, R.E., Glover, W.J. and Schwartz, B.J. (2010). Does the hydrated electron occupy a cavity? Science, 329, 65-69.

ATP and PAPS are unstable in the setting of Inflammation

• pH instability of ATP in low pH range

“…but is rapidly hydrolysed at extreme pH.”

• pH instability of PAPS in low pH range

“…should be made up in pH 8.0 buffer and stored aliquoted at -70o C.”

• Sulfotransferase (SULT) pH optima are typically neutral pH range to slightly alkaline

11

Need for a Universal Sulfurylation Factor in Inflamed Tissue

• A prompt response is required for the GSP/SSP, DIC, Purpura Fulminans

• There is not enough time for de novo protein synthesis and diffusion of substrates, and/or

• To rescue PAPS and ATP from a "hydrolytic fate“ or a recycling, proofreading “mistake”

12

Our Proposal for a Universal Sulfurylation Factor

• Takebayashi, J., Kaji, H., Ichiyama, K., Makino, K., Gohda, E., Yamamoto, I. and Tai, A. (2007). Inhibition of free radical-induced erythrocyte hemolysis by 2-O-substituted ascorbic acid derivatives. Free Radical Biology and Medicine, 43, 1156-1164.• Verlangieri, A.J. and Mumma, R.O. (1973). In vivo sulfation of cholesterol by ascorbic acid 2-sulfate. Atherosclerosis, 17, 37-48.• Cabral, J., Haake, P. and Kessler, K. (1998). Rearrangement of 3-Acyl Derivatives of L-Ascorbic Acid. Journal of Carbohydrate Chemistry, 17, 1321-

1329.

WE PROPOSE: 2-O-sulfate L-ascorbate radical plays a central role in the "universal nonspecific mesenchymal reaction", which has also been referred to as the Sanarelli-Shwartzman phenomenon.

13

• Chiral

• Paramagnetic

• Quasi-aromatic

Need for a Universal Phosphorylation Factor in Inflamed Tissue

• A great need also exists for an ultimate phosphorylation factor in inflamed tissue to

(a) rescue ATP from a "hydrolytic fate“, and

(b) rescue DNA/RNA from proofreading "errors".

14

Proposed TBP TS for sulfuryl and phosphoryl group transfer by Ascorbate derivative radicals

15

“Flattening” the Free Energy Landscape for Biological Group Transfers, generally

• Having proposed chemical biological means by which the Ascorbate radical can catalyze sulfuryl and phosphoryl group transfer, under conditions of inflammatory stress, it soon became apparent that:

• Similar chemical biological means might exist by which the Ascorbate radical can also catalyze nitrosyl, acyl, and glycosyl group transfer, under conditions of inflammatory stress

16

Copper-Ascorbate Complex

17

Source:

L. Amudat, Ph.D.Dissertation, 2010

A NEW PARADIGM FOR HYPOTHESIS TESTING:

• Null hypothesis: The Cu1+ Ascorbate radical complex does NOT directly modulate miRNA synthesis by RNA polymerase III in human patients with cancer, i.e. under conditions of inflammatory stress (low pH and high oxidative stress).

• Application: Disproving the null hypothesis would allow us to assert that the Cu1+ Ascorbate radical complex directly modulates miRNA synthesis by RNA polymerase III in humans with cancer, perhaps as a sequela of a proofreading "error" or a sequela of dysregulating of a mesenchymal stem cell, under conditions of inflammatory stress.

18

Fractones, Defined:

• Fractones have been referred to as stem cell “niches” and they are thought to regulate cytoskeletal assembly and organize the ECM of the heart, gut, brain, and bone marrow/RES. They have complex cytoarchitectures consisting of stem cells, progenitor cells, supporting cells, and laminin-rich basement membranes (Hochman-Mendez et al 2014).

• Frederic Mercier and his associates have studied neural stem cell niches and described fractones as:

“particulate extracellular matrix structures that I previously characterized in both the developing and adult brain”. --- F. Mercier (personal statement) “In the neural stem cell niche of the adult brain, I have demonstrated that fractone-associated heparan sulfate proteoglycans serve as captors and activators of growth factors to regulate neural stem cell proliferation.”

Mercier, F., Kitasako, J.T. and Hatton, G.I. (2002). Anatomy of the brain neurogenic zones revisited: fractones and the fibroblast/macrophage network. J Comp Neurol, 451, 170-188.

19

CNS Fractones Need Sulfur

• GAGs and HSPGs need a UNIVERSAL SULFURYLATION FACTOR, e.g. 2-O-sulfate-L-ascorbate radical might preempt and supercede PAPS/SULTS

• CNS fractones need BOTH ascorbate and sulfur

• HSPGs have been shown to be low in sulfur at autopsy of CNS fractones in Autism

Corley et al (2012); Mercier et al (2012); Meyza et al (2012); Pearson et al (2013); Mercier et al (2011)

20

21

High-dose Intravenous Ascorbate in Critical Care Medicine

• High Dose Intravenous Ascorbate has become mainstream

• Evidence now exists to suggest an important role in both the prevention and management of critical care disease characterized by endothelial dysfunction [Oudemans-van Straaten, 2014; Rodemeister, 2014].

http://www.ccforum.com/content/18/4/460http://www.ccforum.com/content/18/4/461

22

Early Reversal of Endothelial Dysfunction

SOURCE: Oudemans-van Straaten, H.M., Spoelstra-de Man, A.M. and de Waard, M.C. (2014). Vitamin C revisited. Critical care (London, England), 18, 460.

Conclusion:

Ascorbate radical and putative 2-O-substituted L-Ascorbate derivative radicals,

(a) catalyze group transfer reactions, and

(b) modulate redox kinetics.

The catalysis and modulation function for Vitamin C has supramolecular, epigenetic, biophysically-pleiotropic implications which apply to all aspects of medicine.

23

THANK YOU !

24