Posts made by CrumblingCookie

I've started taking 100mg apigenin twice daily.
However, the half-life of apigenin in rodents is reportedly much longer (several days) than in humans (a few hours) so I'm as yet uncertain about the ultimate dosage.

Apigenin, luteolin and chrysin are all congeneric,
with apigenin having 2 hydroxygroups at its B-ring,
luteolin having only one OH-group at its B-ring and
chrysin having none and being the weakest HDACi of those.

• They inhibit HDAC as well as DNMT and also Histone Methyltransferase (HMT), which is distinct from Histone Demethylase LSD1.
• The mechanism for DNMTi is different than the one by EGCG.
• Their DNMTi ist similarly powerful as 5-Aza-dC! Which sounds impressive because Aza is super toxic and irreversebly damaging to DNA.
• They also inhibit in particular the trimethylation of H3K2 through inhibition of EZH2 (Enhancer of zeste homolog 2).

• In our study, DNMT inhibition by plant flavones seems to be due to the binding of flavones at the catalytic binding pocket of DNMTs. These might be different from catechol-containing polyphenols, such as epigallocatechic-3-gallate, in which the noncompetitive inhibition of DNA methylation catalyzed by DNMTs is majorly due to the higher levels of SAM resulting from the catechol-O-methyltransferase-mediated O-methylation of these compounds

• Our results showed that the intercalation of flavones with calf thymus DNA causes major reductions in the intensity of the UV-Vis absorption band characteristics between 260–280 nm, where Apigenin and Luteolin with the 1 and 2 hydroxy groups in the B ring resulted in significant intercalation with DNA. Chrysin (no hydroxyl group in the B ring) resulted in less significant DNA binding.

• Because Apigenin and Luteolin are less water soluble than Chrysin, they are less solvated and consequently susceptible to intercalation within DNA.

Dietary Flavones as Dual Inhibitors of DNA Methyltransferases and Histone Methyltransferases

Overall, apigenin sounds very promising and powerful.
Whilst also being an aromatase inhibitor one one hand the caveat is that it's really exerting estrogenic and progestogenic effects on the other hand. Which seem quite unpredictable.

@Kvirion said:

Yeah, this explains my negative experiences with butyrate a long time ago...

What do you remember about them?
I don't see how exogenous butyrate supplementation fits into Peat's objections as this foregoes the whole endogenous synthesis from bacterial fermentation. The But is not the cause for fermentation with all its associated endotoxin production but an isolated result of it.

@Gaston said:

Possible role of β-hydroxybutyrate in inducing inflammation in alopecia areata

https://onlinelibrary.wiley.com/doi/full/10.1111/exd.15117

• Serum BHB levels were found to be significantly elevated in patients with AA compared with healthy controls, with higher levels correlating with severity of hair loss. BHB treatment increased inflammatory cytokine production in outer root sheath (ORS) cells, mimicking the inflammatory conditions seen in AA.

Hmm. That's a flip side. So the BHB seems to be strongly immunostimulative. Not necessarily merely proinflammatory per se. That leads us down to question the possibilities of what is underlying such (auto)immunological responses in alopecia areata, which may be due to specific or indistinct pathogen infection. And whether tackling such infection is possible rather than feeding or suppressing the immune response against it.

I can't find BHB serum measurements at the standard labs. Knowing those before would of course be nice to figure out which scenario one's in.

@Gaston said:

https://www.bulksupplements.com/products/potassium-bhb

Those are good offers for BHB. Unfortunately they don't sell butyrate or a-KG yet.

@Hearthfire said:

I've seen some stuff about how potassium may be a key to reversing hair loss.

https://www.hairloss-research.org/carnitine-and-hairloss/

Thanks. If I'm grasping it correctly the effects of the potassium only replaces the use of minoxidil: So in all cases where minoxidil proves to be of no benefit there would be no benefits from potassium either.
That and the AA/BHB-connection really points to wanting to first figure out the underlying mechanism of one's hair loss. Or to at least set up a plan of one's own trials as in "if not AA, then...", "if minoxidil, then K" or "if not minoxidil/K, then..."

In Ray Peat's newsletter "Epigenetics, sickness-aging, and changing science" (2014 01), which IMO starts off as a rather tiring albeit not unjustified rant, HDACi emerge as a significant factor against endometriosis and excessive menstruational symptoms (page 5):

• Although every organ seems to be renewed
  (more or less regularly) by the maturation of stem
  cells, the lining of the uterus is interesting because
  it has a monthly cycle, in which stem cells multi-
  ply actively during part of the month, and then
  enter a phase of maturation, with the ability to
  support a pregnancy. In the proliferative phase,
  cells are dominated estrogen, and in the
  maturation phase, by progesterone. During the
  estrogen phase genes are massively silenced by the
  HDAC enzymes, and during the progesterone
  phase, those HDAC enzymes are inhibited.
  Prolonged excessive exposure to estrogen with
  deficient influence of progesterone can lead to the
  development of endometriosis and endometrial
  cancer. The combination of progesterone and
  other HDAC inhibitors is effective in treating
  endometriosis and endometrial cancer, and proba-
  bly in other tumors, such as neuroblastomas (Atif,
  et aI., 2011) and lymphomas. Cyproheptadine,
  known mainly as an antiserotonergic antihista-
  mine, is also an HDAC inhibitor, helpful in
  treating mantle cell lymphoma (Paoluzzi, et al.,
  2009). Aspirin, with an acetylating effect that has
  been considered to be harmful, appears to activate
  a histone, synergizing with a variety of HDAC
  inhibitors to improve the effectiveness of cancer
  treatments.

And for some more Ray Peat blessing fairy dust to the sake of this thread:

• An epigenetic point of view suggests that a
  generalized view of beneficial synergistic effects
  should be considered--things that fundamentally
  support the organism's full development activate
  genes, and are antagonistic to things which funda-
  mentally interfere with that development. Under
  harmful conditions, genes are being silenced in
  the organism's defense, in an organized way, and
  b understanding the nature of that organisation,
  more coherent interventions to protect the organ-
  ism will be possible.
  Radiation, heavy metals, hypoxia, and estro-
  gen excess tend to create excess gene silencing,
  and what they have in common is the creation of
  over-excitable electrons, a reductive and nucleo-
  philic state. The opposing' electronic state,
  electron-withdrawing, typically with one or more
  ketone groups in resonance with one or more
  double bonds as in naphthoquinones (e.g., Inks, et
  aI., 2012) characterizes many of the protective
  substances, for example emodin (found in cascara
  and aloe), curcumin, progesterone, caffeine and
  theophylline.
  Some chemicals known to have protective effects on
  oxidative metabolism, such as short chain
  saturated fatty acids and procaine and procaina-
  mide, are also HDAC inhibitors with a broad
  range of protective effects. Niacinamide (vitamin
  B3) is a powerful HDAC inhibitor; vitamins A
  and D have some synergistic interactions with
  HDAC inhibitors.

• An optimal air pressure, or balance between
  oxygen and carbon dioxide, regular exposure to
  bright light, and foods that supply an appropriate
  balance of amino acids, minerals, vitamins,
  glucose and protective substances, such as HDAC
  inhibitors, would help to support developmental
  plasticity.

There we have Ray Peat endorsement of HDACis.
And the crucial caveat as to why this will remain in the fringes and shadows of "science":

• Many commonly used drugs have unexpected
  harmful epigenetic (degenerative) side-effects.
  Csoka and Szyf have said (2009) "We propose
  that epigenetic side-effects of pharmaceuticals
  may be involved in the etiology of heart disease,
  cancer, neurological and cognitive disorders,
  obesity, diabetes, infertility, and sexual dysfunc-
  tion." Although many researchers are now inter-
  ested in an epigenetic approach, there is no
  assurance that the medical and pharmaceutical
  industry will ever make the adjustment, because
  the most basic assumptions of their science are
  challenged.

Ray Peat's Newsletter "Imprinting and Aging" (2015 07):

• In aging, expression of genes is broadly inhib-
  ited by a general increase in DNA methylation.
  Because of the involvement of similar epigenetic
  modifications of gene expression, aging can be
  thought of as a type of imprinting that involves
  features similar to learned helpless[ness], in which the
  organs and tissues, including the brain, are unable
  to mobilize the energy needed for ordinary
  adaptive processes.

He then mentioned vasopressin, which may be worth a good look on it being influenced by inhibitors of HDAC, DNMT, HMT. He reports that vasopressin also inhibits klotho and increases tissue calfication by increasing phosphate uptake and decreasing klotho-dependent renal phosphate clearance.
He suggested that such vasopressin along with microvesicles or exosomes of stressed cells is being being carried and released systemically [and from human to human, as is actually known], affecting all other cells and tissues of the body, dragging them into the same downturn.

• Stress affects gene methylation to increase production of vasopressin. Antagonists to vasopressin can reverse the learned helplessness produced by stress. Vasopressin is responsible for the intestinal bleeding of stress, causing constriction of the bowel blood vessels, and damages the barrier function of the bowel, and also the blood brain barrier and generally increases vascular leakiness.

Here is why he maybe didn't warm up to SCFAs like butyrate, as he connected their synthesis to the many drawbacks associated with the abundance of gastrointestinal bacterial fermentation:

• The food industry is promoting the use of
  various gums and starches, which are convenient
  thickeners and stabilizers for increasing shelf-life,
  with the argument that the butyric acid produced
  when they are fermented by intestinal bacteria is
  protective. However, intestinal fermentation
  increases systemic and brain serotonin, and the
  short-chain fatty acids can produce a variety of
  inflammatory and cytotoxic effects.

@cs3000 said:

Looked into the effect of sodium butyrate on causing senescence / stopping cells proliferate. it can do that to normal fibroblasts (even tho showed increased wound healing). but effect is selective

I can't remember which study and which HDACi substance but the other day I also read a paper on wound healing and how, in vitro, almost every single crucial stage of cell line and activity suggested impairments of the different stages of wound closure - which overall however didn't materialize as wound healing was improved and "more structured".
I'm still wondering on what to stack or balance HDACi with to also promote proliferation of progenitor cells, though. Maybe aKG? Which may also induce OCT4, SOX2, KLF2 for reversal to pluripontent stem cells (iPSC)?

@Mauritio said:

"...may be exploited to promote human HF growth, metabolism, pigmentation and/or antimicrobial defences."
https://www.morressier.com/o/event/62d558b38a1a1f00195adc16/article/62fa00a7cf455b001a73dd1e

Did you notice the c. x250 increase in dermcidin? I hadn't known about this antimicrobial peptide before but may now start regularly washing or rubbing my face with butyrate solution. Interesting that the But is a metabolite of S. epidermis and is to both feed the hair follice and to thwart skin and follicle infections. The ex vivo nutrient solution was a mere 11mg But per 100ml (0.01%). I'm going to use 500mg per 100ml (.5%).
The skin surface may be just as out of whack as the intestines after any (early life) antibiotic insults. Will add But + S. epidermis body spray to my imaginary list of product innovations.

I've been wondering whether BHB or butyrate would be better energy substrates to hair follices rather than dextrose. I'm thinking preferrably as potassium or magnesium salts.
But alpha-ketoglutaric acid /glutarate may be the energy substrate to prefer (LeeLemonoil had discovered that paper):

Highlights
 • mTOR and AMPK modulation by rapamycin, metformin, and a-KG induces anagen hair growth
 • Autophagy induction is necessary and sufficient for anagen entry and hair growth
 • Autophagy is increased during anagen phase of the natural hair follicle cycle
 • Aged mice fed the autophagy-inducing metabolite a-KB are protected from hair loss

Stimulation of Hair Growth by Small Molecules that Activate Autophagy

It's widely and cheaply available as arginine aKG. I know there are many shampoos etc. with arginine "to promote skin circulation" but not sure if the arginine is a good part. Calcium aKG is difficult and expensive to get and I reckon the calcium is disadvantageous in the skin.
Pure aKG (free acid) is even more difficult to get thatn Ca-aKG. So perhaps the AAKG is good.
a-KB is alpha-ketobutyrate. Which I can't find listed for sale anywhere but it surely looks great for hair.

I've seen apicidin [cyclo(N-O-methyl-l-tryptophanyl-l-isoleucinyl-d-pipecolinyl-l-2-amino-8-oxodecanoyl)] in a graphic put into the same HDACi group of cyclic peptides along with romidepsin and briefly followed up on that one:

It appears apicidin is orally and parenterally effective in vivo against protozae like plasmodium (malaria) by inducing hyperacetylation of histones in treated parasites. Potentially also active against toxoplasma, cryptosporidia etc.

"Other known HDA inhibitors were also evaluated and found to possess antiparasitic activity, suggesting that HDA is an attractive target for the development of novel antiparasitic agents."

Apicidin: A novel antiprotozoal agent that inhibits parasite histone deacetylase

"Treatment of P. falciparum cells with 70nM of apicidin at these three developmental stages resulted in ∼90% reduction of growth (IC90)"

Histone Deacetylases Play a Major Role in the Transcriptional Regulation of the Plasmodium falciparum Life Cycle

This in-silico modelling shows butyric acid being also an inhibitor of class IIa (HDAC7). It also reveals that phenylbutyrate does indeed exert stronger action:

From: In Silico and in Vitro Interactions between Short Chain Fatty Acids and Human Histone Deacetylases

Apigenin and the congeneric luteolin are potent class I HDACi, especially strong against HDAC2 and HDAC8.
Much less affinity to HDAC7, which would be targeted by class IIa HDACi (HDAC4/5/7/9).

[The more negative the value of docking score, the higher is the affinity of ligand toward receptor.]

On a simple by-weight calculation this could convert to 100-150mg apigenin/day for a human:

"Moreover, oral intake of defined flavone (20 and 50 μg/mouse/day) for more than 8-week period has revealed a substantial reduction in tumor growth of PC-3 xenografts in athymic nude mice [41]."

From: In silico approaches for investigating the binding propensity of apigenin and luteolin against class I HDAC isoforms

Luteolin, promising a very good safety profile, may alleviate hyperalgesia, chronic inflammatory or neuropathic pain. The authors make no mention of HDAC or epigenetics at all but point to luteolin's strong anti-inflammatory and antioxidant properties. Ultimately it may well be due to inhibitions of HDAC, similar to and in line with the aforementioned baicalin:

Luteolin: A promising natural agent in management of pain in chronic conditions

@LucyJ said in Tianeptine:

I have a major glutamate anxiety issue and want to try this to see if it helps...

Just a thought if glutamate is at the core of your issues you may need to stabilize the excitory signallings by ramping up intracellular glutathione. E.g. with the help of NAC, plus complementary SFN and low dose B2, B3.
IMO tianeptine is another crutch and good for the first couple of weeks, then quickly diminishing and messy wrt to benefits.

@Mauritio said:

baicalin can ameliorate neuropathic pain by suppressing HDAC1 expression https://pubmed.ncbi.nlm.nih.gov/23684218/

Yet it enhances HDAC2 and inhibits histone demethylase (LSD1) at an IC50 of 3.01μM.
The LSD1 inhibition can be either a welcome or unwelcome dual-function.
https://pubmed.ncbi.nlm.nih.gov/22421405/
https://pubmed.ncbi.nlm.nih.gov/27814566/
I wouldn't know when to use baicalin and when not beside the neuropathic pain scenario.
There has actually been research into a series of dual HDAC and LSD1 inhibitors via conjugating the LSD1 inhibitor tranylcypromine to various o-aminoanilide and hydroxamic acid-based HDAC inhibitors, preferably with vorinostat by Duan et al., 2017 and with entinostat by Kalin et al., 2018.
Domatinostat is by itself already an class I HDACi and LDS1i. Those bifunctional HDACi+LDS1i compounds appear to be significantly more effective against cancer.

Specifically against cancer baicalin works through a different pathway: Inhibition of hexokinase II which is located at the outer mitochondrial membranes and provides the cancer cell's excessive glucose needs.

@alfredoolivas Ah, yes. I'll just wait for the raw powder then to spoon-feed on.
@jamezb46 Thanks for your clarification. I hadn't known about his Arginine Ursolic Acetate specialty. So regular ursolic acid would mainly exert only indirect systemic effects via its intraluminal workings in the GIS. Which may be powerful, but a different ball game.
Have you already tried any of these products like the Ur, 11KT or 11OXO?

@jamezb46
Not about his prohormones, but the other things he sells:
The way he describes ursolic acid (the Ur spray product) makes it appear powerful and wholesome and a much better choice than all that SR9009, RAD140, cardarine etc. stuff.
But if it really contains c. 249mg per recommended daily transdermal dose that is way more than the <4mg daily which Mauritio and cs3000 had figured out, although its effectiveness could perhaps follow an inverted bell curve, and the topical (trans)dermal route may cause less systemic effects:
https://bioenergetic.forum/topic/1797/ursolic-acid-anti-serotonin-anti-estrogen-pro-dopamine-and-more
The raw ingredient is comparatively cheap as I can find 1kg of 98% ursolic acid for about $25. That's 11years of 250mg daily. Or >900years of 3mg daily.
A little surprising to see that he sells BHB(-K?) spray for fat loss. I don't doubt that it could be effective, as I've noticed healing effects by topical butyrate (if mixed into a suitable cream or solution; there are also unsuitable body lotions). Anyhow, that would imply effects predominantly on the skin and subcutanous fat deposits.

@Mauritio said:

Thanks for trying anyways.
That would've been my next step, to look for a device with lower frequency.

"Conflicts of Interest

Authors (M.S., S.K.K., F.M., B.B.R. and R.M.) are co-authors of patents related to these studies, and M.S. and F.M. have financial interests in a company, Mechanobiologics Inc. that is planning to market LFU devices suitable for senescent cell rejuvenation in vitro and in vivo."

Researching the schematics and electrical parts list aside I am wondering how that whole-body treatment could be achieved. In the study they placed the mice on top of a plastic mesh on top of a distancing cylinder in a large beaker filled with water, so that the mice were half-submerged in water and in the far-field of the transducer, i.e. in a relatively homogenous ultrasonic field.
Simply dipping a custom-built transducer at the foot end into the bathtub wouldn't be anywhere as homogenous.
Perhaps place a super-wide-beam transducer at the bottom middle of a bathtub and put a plastic sunlounger on top to lay on. Or integrate it into one of those fancy costly floatation pods.

Does anyone know how to convert those 4kPa power approximately to on-skin mW/cm²?

From looking at the supplemental figure 1 one can see that it's well possible to invest time and effort yet to no or little avail by having the parameters just crucially off: Too much pressure/power (anything above 4kPa) and there's no effect. More on- than off-time pulsing and the effects decrease. A different frequency and there are none of the wanted effects. Too little duration and there's no or little effect yet too frequent repetitions of the same and the effects dimish too.
The supplemental table S2 states the us treatment is strongly antiserotinergic: Peatarians, assemble. Whereas I don't know what to make of those strongly upregulated viral infection processes in table S1.

Very nice find!
You shouldn't have cut off the d) and e) series of mice photographs. Or maybe you purposefully did to tease us, given the many sensations already? Aaaaw, how plump and full of hair those treated mice are in comparison to the fragile control group! And then the videos really emphasize the differences even more. Impressive. Phew.
The treated mice look kind of surreal wrt to that they evidently exhibit the body structure of aged mice but are buffed-up and full of shiny fur.

The most effective ones were the groups that received the least ultrasound

That's cool!

32.2 kHz at 4 kPa for 30 min with a duty cycle of 1.5 s on and 1.5 s off

Bold and original decision and based on original in-vitro findings by the authors!
That's up my alley. Such parameters I could warm up with: Frequencies "just" above the auditory range and long pulses.
-> I've tried to find such an ultrasound device but it seems the I-Tech Mio Sonic from Italy you had found is already on the lowest spectrum and a good one in wide comparison:
Other devices I could find cost at least as much even if looking really cheap and they don't even declare their energy output but it's probably high. They are being pictured for cosmetics use with women using them around their face to treat wrinkles and rub in lotion, pointing them at their temples and sidewards to their eyes. The "professional" devices for medical practitioners costs thousands. They blast up to 3000mW/cm² with a high 150mW/cm² as their absolute bottom setting. The "LIPUS" devices are a subgroup among the devices for medical professionals and are standardized to 1.5MHz, 30mW/cm² pulsed and marketed for fracture healing (for which they are a little controversial, and otherwise on-par with LLLT).
It made me feel all nauseous, really.
It seems such a device has to be built by oneself.

Looking through the graphs in this 1999 paper doesn't definitely answer any of my thoughts but pinpoints further questions I'm having.
They used 45kHz continuous vs the usual 1MHz 1:4 pulsed ultrasound. The various protein expressions from the continous 45kHz are sometimes non-linear to the energy intensity - in contrast to the pulsed 1MHz where the dose-dependent effect appear very linear.

-> Continuous US may be inferior to pulsed US. The tissues may need those resting phases between US bursts.

What's also super evident is that with the long-wave 45kHz, even after multiplying the energy intensities by x5 for equivalent joules, all the cell proliferation maxima are far lower than with 1MHz pulsed: At 5-30mW/cm² (x5 = 25-150mW/cm² pulsed) versus 100-700mW/cm².

On page 7 and 8 are the graphs for FGFb and VEGF expression from monocytes and osteoblasts and those drop off really sharply at anything but the lowest doses beyond 5 or 15mW in the 45kHz cont or beyond 100mW in the 1MHz pulsed.
FGFb (Basic fibroblast growth factor) is even strongly suppressed below control levels at more than 100mW at 1MHz pulsed.

-> Variations in energy intensity, frequency and pulsing definitely induce very opposing effects on respective cellular functions. E.g. stimulating vascularization and noncollagenous protein synthesis takes place at much lower doses than the maxima for fibroblast and osteoblast proliferation.

-> Seeing, however, that even for the latter the effects by far exceeded the controls at the lowest intensities tested (5mW/cm² 45kHz cont and 100mW/cm² 1MHz pulsed) there appears to be no specific incentive to go beyond the minimum dosage in any case, based on this in vitro study.

In vitro effects of therapeutic ultrasound on cell proliferation, protein synthesis, and cytokine production by human fibroblasts, osteoblasts, and monocytes

I am wondering also on the impact of a very low pulse rate (as the 1Hz in the wound healing study above). Maybe that kind of emulates LESWT but I'm no biophysicist.

@Mauritio said:

https://pubmed.ncbi.nlm.nih.gov/35999130/

Yes quite a lot in comparison; 300mW/cm², 5 minutes each on all four sides:

Treatment

LIPUS therapy adopts a LIPUS therapy device (WBL-ED) with a pulse duration time to pulse rest time ratio of 1:4 (200:800 ms) at 1,000 Hz and a frequency of 1.7 MHz (Wan-beili Medical Devices, Beijing, China). An F-type probe and energy level III (equivalent to 0.3 W/cm 2 ) were selected. The skin and probe were evenly coated with ultrasonic couplant. At each treatment session, LIPUS was applied on each side of the penile shaft and crus in turn, 5 minutes per site, for 20 minutes.

The response rates of the 3/W and 2/W groups increased gradually at weeks 4, 6, 8, and 12 in both the FAS and PPS, respectively. The response rates at week 12 in 3/W and 2/W groups were 62.0% and 62.5%, respectively, in the FAS. The response rates at week 12 in the 3/W and 2/W groups were 67.9% and 66.9%, respectively, in the PPS.

The percentage of patients with EHS ≥3 demonstrated significant improvements from baseline to week 12 in the 3/W group (54.9%−84.0%) and 2/W group (59.5%−83.5%),

We could hypothesize that next time they may find 2x/week with <100mW/cm² working similarly well or better not only wrt perfusion but retained/improved composition, maturation and structure of collagen (although that's a difficult thing to observe because to sacrifice and to dissect after experimentation is clearly excluded).

The Johns 2002 paper goes into many depths:

"Specifically, ultrasound has been shown to modulate vasoconstriction; lymphocyte adhesion properties of endothelium, mast cell degranulation, phagocytosis by macrophage, production of growth factors by macrophages; calcium fluxes in fibroblasts; angiogenesis; proliferation of T-cells, osteoblasts, fibroblasts, and a number of proteins associated with inflammation and repair (IL-1, IL-2, IL-6, IL-8, interferon-γ, fibroblast growth factor-b, vascular endothelial growth factor, collagen) (Table)1,34,40–42,45–53; and to accelerate thrombolyisis.7–16

In general, most of these researchers used a frequency of 1 MHz or 3 MHz, and the intensities ranged from 0.1 to 1.5 W/cm2.

An alternative therapeutic protocol employs a frequency of 45 kHz. An intensity range of 5 to 100 mW/cm2 was shown to increase the production of IL-1, IL-8, vascular endothelial growth factor, fibroblast growth factor-b, and collagen; promote bone healing; and accelerate thrombolysis.5,45,54,55

The long-wave (45-kHz) ultrasound increases penetration depth and, therefore, seems to be more appropriate than traditional high-frequency ultrasound (1 MHz and 3 MHz) for promoting revascularization and bone healing at greater depths."

"The frequency resonance hypothesis suggests that the energy provided to the enzyme by the ultrasound wave may induce transient conformational shifts in certain enzymatic proteins, altering the enzyme's activity (ie, kinases or phosphatases) and the overall function of the cell (Figure 1). Alternatively, ultrasound's resonating force may result in the dissociation of functional multimolecular complexes (Figure 2) or the release of a sequestered molecule by dislodging an inhibitor molecule from the multimolecular complex (Figure 3). In essence, the mechanism of ultrasound's action in Figures 2 and 3 is the same. "

"Hypothetically, frequency resonance may imply that different frequencies (1 MHz, 3 MHz, 45 kHz, and others) establish unique resonant or shearing forces (or both). Moreover, various frequencies may affect combinations of proteins or multimolecular complexes in different ways, lending to the possibility of targeted effects at the cellular and molecular levels."