Chapter 13: Swelling, Shrinkage, and Volume Control of Living Cells

ATP controls cellular volume - a different look at "leaky gut and the blood-brain barrier."

As always, this is not medical advice, and reading this does not form a client relationship with me - your health is your responsibility.

Many have asked for my “thoughts” on things. The best place to start would be some of my older Substacks - especially the ones on supplementation (1 and 2). Also, please check out my IG story highlights.

Today’s Substack will continue with Chapter 13, Swelling, Shrinkage, and Volume Control of Living Cells. This is one of my favorite chapters as I think it is behind much of the “leaky gut and blood-brain barrier” and edema issues many face. Chapter 12 discussed cell permeability to different things, including water and glucose.

Summary:

The AI hypothesis centers around - the adjustment “of the activity of cell water, so that it remains in equilibrium with that of water in the external medium, is a result of the polarization of water in multilayers by a matrix of extended cell proteins.” ATP extends these cell proteins via proper folding and unfolding. The “two major factors that determine cell volume: (1) the degree of polarization or depolarization of cell water and (2) the formation or release of inter- and intramolecular salt linkages.” “One of the major functions of ATP is to maintain cell volume, and it does this via three mechanisms: (1) it potentiates polarization of water in multilayers, (2) it potentiates adsorption of a cation (usually K+) onto fixed anionic sites that could potentially form salt linkages with fixed cationic sites, and (3) it tends to maintain preexisting salt linkages that are otherwise susceptible to breakage by the usual high concentration Na+ in the external medium. The effects of ATP depletion are therefore (1) a decrease in the number of adsorbed cations (i.e., a decreased K+ that is not replaced by Na+),” (2) less dissolved solutes in cell water, “and (3) a tendency of NaCl to break open preexisting salt linkages. Cells depleted of ATP tend to swell.”

13.1 The Refutation of the Membrane Theory of Cell Volume Regulation

The cell membrane view requires a continuous and intact cell membrane like a water balloon with little toys floating inside. Once the balloon is cut, the volume will go to zero, and the constituents will spew out. If cells operated this way, the same thing would occur if a cut, etc., was applied to a cell membrane. This is not seen - e.g., cell lysis, etc. According to Ling, “the maintenance of cell volume in a normal or a swollen state does not even require an intact cell membrane.”

13.2 Polarized Water in Lieu of Free Intracellular K+ in the Maintenance of Osmotic Pressure of Living Cells

Living cells have the remarkable ability to maintain their size and shape, even when exposed to solutions with varying levels of solutes. This means they can swell when placed in a solution with fewer solutes (hypotonic) or shrink when in a solution with more solutes (hypertonic), all without needing a traditional cell membrane to keep them intact. “Living cell surfaces are, with no known exception, permeable to water. Net movement of water from the cell interior to the outside indicates a difference in the activity of water in the two phases. For example, introduction of normal cells into a hypertonic external solution, which has a lower activity of water, leads to loss of water from the cell.”

13.3 What Does the Vapor Sorption Isotherm Tell Us about the Osmotic Behavior of Living Cells?

According to the membrane theory, scientists expected that the water content of living cells would follow a well-known equation called the van't Hoff equation, which relates osmotic pressure (a measure of the cell's response to solute concentrations), the volume of water associated with solutes in the cell, temperature, and gas constant. Experiments done by Ling and others showed that the linear relationship between volume changes and osmotic pressure in conventional studies was coincidental. As water activity approached zero, cells did not retain 30% of their water as previously believed; instead, they lost all of it - this makes sense, given societies preserve food via drying it. Water activity is a measure of how much water is available for osmosis to occur. It plays a central role in determining the direction and strength of osmotic processes. It is equal to the partial vapor pressure (=p/p0 where p is the vapor pressure of water in solution and p0 is the vapor pressure of pure water at the same temperature and pressure).

13.4 Swelling of Living Cells in Isotonic KCI and Other Salt Solutions

Multiple studies looked at the behavior of frog muscle cells in response to different concentrations of potassium chloride (KCl) and chloride ions (Cl-) in their surrounding environment. This research showed that KCl can cause swelling in these cells.

1. Initially, it was believed that frog muscles remained unchanged when placed in isotonic solutions of sodium chloride (NaCl) but swelled when exposed to KCl. This was thought to be due to the perceived impermeability of NaCl and the permeability of KCl. However, it was later understood that NaCl and KCl are permeable to the cells, and the explanation was more complex.

2. According to the AI hypothesis, the resting frog muscle cells would have taken up more water without salt linkages between protein chains. Salt linkages involve interactions between cationic groups (e.g., amino groups from lysine and guanidyl groups from arginine and other amino acids) and anionic groups (e.g., carboxyl groups from aspartic and glutamic acid residues) within proteins that the water, etc. are adsorbed on. These linkages play a role in restraining the cells from taking up excessive water and, when dissociated, contribute to cell swelling.

3. The salt linkages restrict the swelling of muscle cells, but when exposed to high concentrations of KCl, many of these linkages are dissociated (=broken). This allows the cells to pick up more water, leading to swelling. The extent of swelling depends on the concentration of extracellular KCl, with certain concentrations causing stepwise vs. continuous swelling - most of nature is quantized, these stepwise processes are not only seen in quantum mechanics.

4. Experiments involving frog muscle cells exposed to KCl solutions revealed different fractions of Cl- efflux with varying half-lives, corresponding to different types of bound Cl-. The results matched theoretical predictions based on salt linkage dissociation and Cl- binding.

5. The swelling of muscle cells is closely related to the dissociation of salt linkages and the creation of new adsorption sites for both K+ and Cl-. The experimental results aligned with theoretical expectations, indicating a strong link between Cl- efflux and the swelling behavior observed in frog muscle cells.

13.6 The Mechanism of Cell Swelling Caused by the Depletion of ATP and the Role of NaCl in the Medium

“Salt linkages between protein chains play a role in the maintenance of the normal cell shape.” These cationic and anionic sites are “under the control of the cardinal adsorbent ATP.” As such, “one can expect that the volume (size) and shape of living cells will be dependent on ATP.” This is arguably the most important concept I have ever encountered in health - if the volume and shape of living cells depend on ATP, every cell action is. The first time I read the preceding passage (found on page 455 in Ling’s book), the collective weight of the alternative health world I had been obsessively and neurotically researching lifted off my shoulders as I finally knew I had found my “Pareto principle.” This may seem like a “no duh, ATP is the important idea” to most. However, before this, I was in every rabbit hole (e.g., EBV, fasting, carnivore, mono-fruit, lyme, parasites, adrenals, the list goes on a while) trying to figure out what was “wrong” with me and more so what to do about it. This idea washed all that away and directed my efforts. In doing so, I healed a laundry list of diagnoses and have been able to help many overcome theirs since.

Examples of ATP’s relationship to size and shape:

1. “Rabbit psoas muscle fiber bundles abruptly shortened in an "all-or-none" manner when their ATP content fell below 50% of its normal value.” - important for those with muscle twitching, hyper-/hypotonic muscles, rigor mortis, etc. Thankfully, we have fatigue mechanisms to limit ATP levels from falling too much. However, that does not mean one is not on the tail of this normal or signaling curve.

2. Human erythrocytes shape and size depend “on their ATP content when the cells were suspended in physiological saline to which was added glycolytic poison, NaF.”

3. Muscle swelling in death - why edema is not a “good” sign, ascites experienced by many who are dying, lymphedema in cancer, etc.

According to the AI hypothesis, “ATP adsorption at the cardinal sites maintains the anionic side chains of the involved proteins.” This leads to “K+, Rb+, NH4+ and the fixed cations (alpha-amino, epsilon-amino, and guanidyl groups) are preferred over Na+, Li+, or Cs+. “Loss of ATP… [causes] the relative affinity for Na+ [to increase] and that for fixed cations decreases.” This is why I have referred to Na+ as a “band-aid” for a low ATP state. If anything, the Na+ is helping to maintain the cellular potential in a low ATP state. This is why, while salt is important, the transition to needing a ton of it can indicate health decline. Li+ can substitute for Na+ in this state, too - interesting considerations for mental health.

There is an initial increase of intracellular water “during the initial fall of ATP content and then rapidly accelerated [increase] with [the] disappearance of the last [of the ATP].”

The mechanism of cell swelling due to hypotonic solutions, isotonic KCl, and “from ATP depletion brought on by poisons or cold temperatures” are different. The ATP depletion case sees an “accumulation of loosely held normal water” that is readily excreted (=death).

13.7 Classification of Cell and Tissue Swelling

The three major types of cell swelling:

1. Osmotic swelling/ shrinkage - living cells transferred from a concentrated to a more dilute solution, dependent on water activity.

2. Dissociative swelling - caused by the dissociation of salt linkages and depends on the properties of both cations and anions in the salt. / associative shrinkage - swelling induced by isotonic NaCl in ATP-depleted cells, leading to cell water depolarization.

3. Desorptive swelling - occurs in dying cells, typically before ATP is entirely exhausted; cell K+ desorbs before water is depolarized. The reduction in water activity due to free K+ induces swelling.