2026-08-11
Advantage to integrate movement (Yoga or Chi Kung inner exercises), massage with sound resonance on slow exhalation breathing
AI-översikt:
Integrating slow exhalation breathing with movement, internal exercises (like Yoga or Chi Kung), and sound resonance creates a powerful somatic synergy. This combination rapidly triggers the vagus nerve, enhances internal cellular micro-massage through sound vibrations, shifts the nervous system into deep recovery, and optimizes cardiovascular coherence.
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Nervous System Regulation
- Parasympathetic activation: Extended, slow exhalations signal the brain to switch off the ”fight or flight” response and engage ”rest and digest” mode.
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- Vagal stimulation: The mechanical movement of the diaphragm combined with vocalized sound resonance physically massages the vagus nerve pathways in the chest and throat.
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- Heart rate variability (HRV): Slow, rhythmic breathing synchronizes cardiac and respiratory rhythms, improving the body’s internal feedback loops.
Ohm Health
BvS-> HRV is spectral analysis of heart rate time series and Respiratory Sinus Arrhythmia (RSA) is Autonomic Nervous System (ANS) oscillatory pattern – breathing out inhibit sympathetic part) activity opening for PNS (Parasympathetic part) and reversed
Physical and Energetic Integration
- Internal micro-massage: Sound resonance (such as humming or toning on the exhale) sends gentle pressure waves through tissue, easing structural tension.
SoundSelf
- Fluid movement: Combining Yoga or Chi Kung movement with a prolonged outbreath helps release stubborn muscular holding patterns and joint stiffness.
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- Enhanced circulation: Paced movements paired with regulated breathing support optimal blood flow and efficient gas exchange in the lungs.
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Mental and Emotional Clarity
- Mind-body anchor: The dual focus on physical form and sound gives an overactive mind a stable point of rest, deepening meditative states.
- Stress dissipation: The rhythmic loop reduces baseline psychological tension and lowers stress hormone production.
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I’ll tailor my next response. Just tell me:
- Chi Kung or Yoga primary practice focus
- Specific sound type (humming, toning, or silent)
- Target goal (sleep, stress relief, or energy)
BvS-> The main problem right now is
-> How can I create sound resonance which influence facilitate the creation of optimal enzymatic influence on mitochondria?
AI-suggest
Creating direct sound resonance to specifically optimize individual mitochondrial enzymes inside living human tissue remains an experimental theoretical concept rather than an established clinical practice. While emerging in-vitro studies suggest that specific acoustic waves and musical frequencies can influence cellular bioenergetics, mechanotransduction, and ATP production, targeting distinct internal enzymatic reactions via external sound requires precise physical parameters. [1, 2, 3]
- Cytoskeletal Transmission: Vibrations transfer through the cytoskeleton directly to organelles, altering mitochondrial trafficking and membrane oscillations. [1]
- Pulsed Ultrasound: Non-thermal low-intensity ultrasound parameters (e.g., specific MHz or kHz ranges delivered at low duty cycles) are researched for their ability to safely stimulate metabolic pathways without causing thermal damage. [1]
- Vibrational Coherence: Theoretical models suggest that matching the natural oscillatory frequencies of cellular structures may promote synchronization of membrane potentials, though reproducible parameters for specific mitochondrial enzymes (like ATP synthase) are not yet standardized for human application. []How can I create sound resonance which influence facilitate the creation of optimal enzymatic influence on mitochondria?
https://wms-site.com/alert-on-mitochondria/1154-mitochondrial-oscillations-concept-and-future-perspectives
Exploring Mitochondrial Oscillation and Resonance: Impact on ATP Production and Distribution in Health and Diseases
- Details
- Published on 13 July 2023
What do we mean by Mitochondrial oscillations and resonance?
Mitochondrial oscillations and resonancecould indeed have significant implications for ATP production and distribution. As the primary ”powerhouses” of the cell, mitochondria are responsible for the bulk of ATP production through the process of oxidative phosphorylation. The efficiency of this process is intimately linked to the mitochondrial membrane potential, which undergoes periodic oscillations.
These oscillations can influence the rate of ATP production in a couple of ways. When the membrane potential is high, the mitochondria are in an energy-conserving state, and ATP production is typically efficient. Conversely, when the membrane potential is low, the mitochondria are in an energy-dissipating state, and ATP production may be less efficient.
In terms of ATP distribution, mitochondria are known to form dynamic networks within cells, allowing for the efficient transfer of ATP and other metabolites to where they are most needed. The oscillatory behavior of mitochondria could potentially play a role in regulating this distribution process. For instance, changes in the oscillatory state of a particular mitochondrion could trigger the redistribution of ATP within the mitochondrial network, allowing the cell to rapidly respond to changing energy demands.
Moreover, the concept of mitochondrial resonance could further impact ATP distribution. If different mitochondria within a cell or tissue can ’resonate’ with each other – that is, synchronize their oscillations – this might enable a more coordinated and efficient distribution of ATP throughout the cell or tissue.
The concept of mitochondrial resonance is a fascinating one and may have far-reaching implications for how energy is distributed within cells and tissues.
As an analogy, consider the way synchronized swimmers can move more efficiently through water by coordinating their movements. Similarly, if different mitochondria within a cell or tissue can ’resonate’ with each other – that is, synchronize their oscillations – this might enable a more coordinated and efficient distribution of ATP throughout the cell or tissue.
When mitochondria are in resonance, the oscillations of their membrane potentials are synchronized. This synchronization might facilitate the formation of a more efficient energy distribution network, allowing ATP and other metabolites to be rapidly and efficiently transported to where they are most needed within the cell or tissue.
This could be especially important in cells with high energy demands, such as neurons or muscle cells, or in situations where energy needs can change rapidly, such as during exercise or in response to stress. Resonance might allow these cells to quickly ramp up ATP production and distribution in response to increased energy demand, ensuring that all parts of the cell have access to the energy they need to function optimally.
Mitochondrial Oscillation – Project’s Aim
Our goal is to probe deeper into the complex biological phenomena of mitochondrial dynamics – oscillations (continuous changes in mitochondrial membrane potential) and resonance (amplification of effects of an external force matching a system’s own natural frequency) – and their influence on the cellular energy landscape.These oscillations play a pivotal role in cellular signaling, energy metabolism, and reactive oxygen species production, with alterations linked to various pathological conditions such as neurodegenerative diseases and cancer. Moreover, mitochondrial resonance potentially enhances inter-mitochondrial and cellular communication, optimizing energy transfer and signaling pathways.
A key aspect of this project focuses on the implications of these processes for ATP production and distribution. Mitochondria, as cellular powerhouses, are primarily responsible for ATP production via oxidative phosphorylation.
This project will explore:
- How changes in mitochondrial membrane potential during oscillations influence the efficiency of ATP production.
- How mitochondrial network dynamics, regulated by oscillatory behavior, can impact ATP distribution within the cell.
- How mitochondrial resonance may enable a more coordinated and efficient distribution of ATP throughout cells or tissues. The concept of mitochondrial resonance could further impact ATP distribution. If different mitochondria within a cell or tissue can ’resonate’ with each other – that is, synchronize their oscillations – this might enable a more coordinated and efficient distribution of ATP throughout the cell or tissue.
Abstract
Background
Pulsed ultrasound (US) is widely used both as a diagnostic imaging tool and a therapeutic approach. However, many of the mechanisms underlying the therapeutic effects of non-thermal US remain unclear, especially in skeletal muscles, which play a crucial role in the body’s metabolism. The aim of this study was to investigate the effects of US on myotubes.
Methods
In this study, C2C12 myoblasts were utilized. After differentiating into myotubes, the cells were exposed to US irradiation at an intensity of 3.0 W/cm2, with a 20 % duty cycle, an acoustic frequency of 1 MHz, and a pulse repetition frequency of 100 Hz for 5 min. The cells were then collected and analyzed for genomic and metabolomic alterations, as well as mitochondrial function.
Results
Cell viability remained unaffected after US irradiation. The mitogen-activated protein kinase (MAPK) signaling pathway was the most activated, while the expression of various RNAs was significantly altered. Purine metabolism was highly activated, with an increase in the abundance of metabolites associated with this pathway. Furthermore, mitochondrial respiration in the myotubes increased following US irradiation.
Conclusion
This study investigated the impact of US irradiation on myotubes using genomic analysis, metabolomic analysis, and mitochondrial function. US irradiation activated the MAPK signaling pathway, which in turn enhanced purine metabolism and improved mitochondrial respiration.
Keywords: Myotubes, Pulsed ultrasound, MAPK signaling pathway, Purine metabolism, Mitochondrial respiration Highlights
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Ultrasound irradiation activated purine metabolism without increasing the expression of uric acid.
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Ultrasound irradiation increased mitochondrial respiration in myotubes.
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Ultrasound irradiation activated the MAPK signaling pathway in myotubes.
1. Introduction
Pulsed ultrasound (US) is extensively utilized not only for diagnostic imaging but also as a non-invasive, painless, and safe therapeutic approach, rendering it suitable for a diverse range of patients [1]. With a long history, the non-thermal impacts of US have attracted increasing attention [2]. Its applications in alleviating pain, promoting tissue healing, and improving tissue characteristics have grown in popularity [[1], [2], [3]]. Therapeutic US is commonly used to treat muscle injuries [4], has anti-inflammatory effects [5], and prevents muscle atrophy [6]. Despite the plethora of potential therapeutic applications, the mechanisms underlying the non-thermal effects of US remain largely unexplored and are still under investigation [2].
Skeletal muscle is the largest organ in the human body, accounting for about 40 % of total body mass [7,8]. It contributes approximately 30 % to the resting metabolic rate and is implicated in up to 75 % of total body metabolism [9]. It plays a crucial role in glucose uptake and is pivotal in both exercise and metabolic disease [10]. Skeletal muscle is also vital for regulating its own metabolism, particularly in response to exercise and pathological conditions [[11], [12], [13]]. The mitogen-activated protein kinase (MAPK) signaling pathway plays an essential role in cellular metabolism [14]. Mitochondrial function contributes significantly to muscle metabolism [15]. Mitochondria are essential organelles in cellular metabolism that play a key role in regulating the metabolic state of skeletal muscles [15,16]. Therefore, studying the intricate roles of the MAPK signaling pathway and mitochondrial function in skeletal muscle metabolism is vital for advancing our knowledge of metabolic regulation and potential therapeutic interventions.
As US irradiation regulates the MAPK pathway [17], which influences skeletal muscle metabolic alterations, we hypothesized that US regulates the skeletal muscle metabolic state through the MAPK signaling pathway. Given the unclear effects of US irradiation on metabolites and their mechanisms in skeletal muscle, this study aimed to determine the effects of US on myotubes using genomic analysis, metabolomic analysis, and mitochondrial function.
2. Materials and methods
2.1. Cell culture
C2C12 myoblasts, mouse skeletal muscle cells, were obtained from the American Type Culture Collection (ATCC). These cells were cultured in 35-mm dishes containing Dulbecco’s modified Eagle medium (DMEM) supplemented with 10 % fetal bovine serum (FBS) at 37 °C under 5 % CO2. The growth medium was replaced with a differentiation medium consisting of DMEM supplemented with 2 % horse serum (HS) at approximately 90 % confluence. The cells were cultured for 6 or 7 days to allow differentiation. After differentiation, US irradiation was applied to the myotubes.
2.2. US irradiation
Following cell differentiation, US irradiation was performed for 5 min using a medical US device (SZ-100 M; MINATO Medical Science 129 Co., LTD, Japan). The device probe was placed beneath the bottom of the culture dish, and a coupling gel was used between the culture dish and probe. A piece of sterilized silicone was suspended 2 mm above the cell monolayer in culture media [18]. US parameters were as follows: intensity, 3.0 W/cm2; duty cycle, 20 %; acoustic frequency, 1 MHz; and repetition frequency, 100 Hz. The beam nonuniformity ratio, which represents the ratio of the maximum intensity to the average intensity of the US, was 2.4, confirming that the probe was safe. The temperature of the culture medium was below 37 °C during the US irradiation.
2.3. Cell viability analysis
In this study, myotube viability was evaluated using the MTT assay after US irradiation. Cells were incubated with 5 mg/mL MTT solution (10 × ) and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2 H-tetrazolium bromide (Wako Junyaku Co., Ltd., Japan) diluted in the culture medium. Following incubation for 3 h, dimethyl sulfoxide (DMSO) was added. The absorbance was measured at 595 nm using an MTP-300 spectrophotometer (Kono Electric Co., Ltd., Japan). Cell viability was expressed as a percentage relative to the CON group.
Myotube viability was also evaluated using Zombie Red™ staining. The cells were rinsed twice with PBS and incubated with the Zombie Red™ Fixable Viability Kit (1:1000; BioLegend, CA, USA) for 15 min. Zombie Red™ is an amine-responsive fluorescent dye that is impermeable to living cells but permeable to those with damaged plasma membranes. Following incubation, the cells were rinsed twice with PBS and incubated with 4 % formaldehyde at room temperature for 30 min. The cell nuclei were counterstained with DAPI (1:1, 000; Dojindo, Kumamoto, Japan) for 5 min. The stained cells were imaged using a fluorescence microscope (BZ-X800; Keyence, Japan).
2.4. Reactive oxygen species (ROS) analysis and DNA damage analysis
ROS production was analyzed using CellRox Green Reagent (Invitrogen). The reagent was added to each dish at a concentration of 10 μmol/L, and the cells were incubated for 30 min at 37 °C in a 5 % CO2 incubator. The dishes were washed with PBS and incubated with 3.7 % formaldehyde for 15 min at room temperature. DAPI (1:1000; Dojindo) was added, and the cells were incubated for 5 min at 37 °C after washing with PBS. Stained cells were observed under a fluorescence microscope (BZ-X800; Keyence, Japan). The fluorescence intensity of each dish was determined using Image J, and the results were analyzed as a ratio to those of the control group.
TUNEL staining was performed using an in situ Apoptosis Detection kit (Takara In Situ Apoptosis Detection Kit; Takara Bio Inc., Shiga, Japan) to analyze DNA damage in the cells. The cells were then rinsed twice with PBS and incubated with 4 % formaldehyde at room temperature for 30 min. After rinsing the cells twice with PBS, 0.3 % H2O2 in methanol was added at room temperature. After 30 min, cells were washed twice with PBS, and permeabilization buffer was added for 5 min at 4 °C. The TdT enzyme and labeling safe buffer were then added and incubated at 37 °C for 60 min. The cell nuclei were counterstained with DAPI (1:1, 000; Dojindo, Kumamoto, Japan) for 5 min. The stained cells were imaged using a fluorescence microscope (BZ-X800; Keyence, Japan).
2.5. Seahorse assay
To evaluate the bioenergetic condition of the myotubes after US irradiation, a Seahorse XFp Analyzer (Agilent Technologies, USA) was utilized for mitochondrial stress tests to measure the oxygen consumption rate (OCR). C2C12 myoblasts were cultured in XFp cell culture miniplates to differentiate into myotubes. After differentiation, the myotubes were subjected to US irradiation for 5 min. The Seahorse assay was then performed after a 24-h period.
For OCR detection, the medium was replaced with Seahorse XF Assay medium supplemented with 10 mM glucose, 1 mM pyruvate, and 2 mM glutamine 24 h after US irradiation. The miniplate was then incubated at 37 °C in a non-CO2 incubator for 30 min. OCRs were measured under basal conditions and after sequential application of 1 μM oligomycin (to assess ATP-related oxygen consumption), 1.5 μM carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) (to determine the maximum respiratory capacity), and 2 μM rotenone/antimycin A (R/A) mixture (to evaluate the non-mitochondrial oxygen consumption). After OCR was detected, the cells were rinsed once with PBS and lysed in lysis buffer. The lysate was centrifuged to obtain the protein supernatant. The BCA assay was employed to measure the protein concentration. OCR values were normalized to protein concentration and expressed as pmol/min/μg protein.
2.6. RNA sequencing of myotubes
Total RNA was extracted from the myotubes using TRIzol reagent (Takara Biotechnology, Japan). Raw RNA sequence data were obtained utilizing an Illumina NovaSeq 6000 machine. The fold change (mean expression of each RNA in the US group divided by the mean expression in the control group) and P value for each RNA were calculated after obtaining the raw data. These P values were then used to determine the false discovery rate (FDR) for each RNA, which was then applied as a filter to identify essential RNAs with an FDR <0.05. RaNA-Seq and the R 3.5.3 program were utilized for pathway enrichment analysis. After performing pathway analysis using the Kyoto Encyclopedia of Genes and Genomes database, the 10 most enriched pathways associated with signal transduction were identified and used to explore related pathways. The RNA-seq datasets produced in this study have been uploaded to the NCBI Sequence Read Archive. The accession number for the dataset is PRJNA1116252. Detailed sequencing data and relevant metadata can be accessed at https://www.ncbi.nlm.nih.gov/search/all/?term=PRJNA1116252.
2.7. Metabolite analysis
At 24 h after US irradiation, myotubes were rinsed twice with PBS and then lysed in 80 % methanol containing 50 μM (+)-10-camphorsulfonic acid, 400 μM l-methionine sulfone, and 400 μM piperazine-1,4-bis (2-ethanesulfonic acid) as internal standards. The cells were incubated at −80 °C for 15 min followed by scraping and centrifugation at 14,000 g for 5 min at 4 °C. The supernatants were collected and filtered through a Millipore 5 kDa cut-off membrane to remove solubilized proteins. The dried metabolites were dissolved in Milli-Q water after evaporating the aqueous layer extracts under vacuum using a FreeZone 2.5 Plus freeze-dry system (Labconco, Kansas City, MO). Intracellular metabolite concentrations were analyzed using LC-MS to detect metabolites from a library of 300 targeted compounds, including those in the TCA cycle, 3-phosphoglycerol shuttle, and glycolysis [19]. MetaboAnalyst 5.0, an online software, was used for pathway analysis (https://www.metaboanalyst.ca) [20].
2.8. Western blotting
The culture medium was removed, and C2C12 myotubes were washed with PBS, scraped, and homogenized in lysis buffer. Homogenates were centrifuged at 15,000 rpm for 10 min at 4 °C, and the supernatants were collected. The supernatant protein concentrations were determined using a BCA protein assay. Samples were standardized, and equal amounts of protein were separated through 7.5 % or 12.5 % SDS-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes. The membranes were blocked with 3 % BSA in TBS with 0.1 % Tween 20 (TBST) or 5 % skim milk in PBS with 0.1 % Tween 20 (PBST). Membranes were incubated with p38 MAPK (1:1000 in TBST, #8690, Cell Signaling), phospho-p38 MAPK (1:1000 in TBST, #4511, Cell Signaling), and GAPDH (1:1000 in PBST, #97166, Cell Signaling) at 4 °C overnight. The membranes were then incubated with horseradish peroxidase-conjugated anti-rabbit or anti-mouse IgG (1:10,000 in TBST; GE Healthcare, Waukesha, WI, USA) for 1 h at room temperature. Membrane proteins were detected using a chemiluminescent reagent (Ez West Lumi; ATTO, Tokyo, Japan) and quantified with an image reader (LAS-1000; Fujifilm, Tokyo, Japan).
2.9. Statistical analysis
All values are expressed as means ± standard error of the mean (SEM). Differences were considered statistically significant when p < .05, as determined by a two-tailed Student’s t-test.
3. Results
3.1. US irradiation does not affect cell viability
As depicted in Fig. 1A, compared to that in the control group, there was no decrease in cell viability in the US group, suggesting that US irradiation did not reduce cell viability. Additionally, as depicted in Fig. 1B and C, Zombie Red™ immunofluorescence staining was performed to evaluate the cytotoxic effects of US irradiation on myotubes. Treatment with 1 % povidone-iodine significantly reduced cell viability, while US irradiation did not affect cell viability. In addition, we verified whether US irradiation increased ROS expression in myotubes through immunofluorescence staining for ROS and whether it caused DNA damage in myotubes using TUNEL staining. The results showed that US irradiation did not increase ROS expression in myotubes (Supplementary File, Fig. 1) or cause DNA damage (Supplementary File, Fig. 3).
Fig. 1.
Cell viability. A: MTT assay was performed after ultrasound (US) irradiation to measure cell viability, expressed as a percentage relative to the control. All values are represented as mean ± SEM (n = 3). B: Zombie Red™ immunofluorescence staining (red) in myotubes after US irradiation. After fixation, myotubes were counter-stained with DAPI (blue). Scale bar = 100 μm. C: Cell viability analysis using Zombie Red™ immunofluorescence staining. Mean fluorescence intensity of Zombie Red™ in myotubes. Values are presented as percentage of povidone-iodine and expressed as means ± SEM. Fig. 3.
Effect of US irradiation on p38 MAPK in myotubes. A: Representative western blots of Phospho-p38, Total-p38 and GAPDH. B: The phosphorylation and total protein levels of p38 MAPK were measured in this study. 3.2. US irradiation most effectively activated the MAPK signaling pathway
RNA sequencing was performed to determine the mechanisms activated by US irradiation in myotubes after US irradiation. A total of 13,660 RNAs were identified using quantitative proteomic analysis. Differentially expressed RNAs were visualized using a volcano plot (Fig. 2A). Two hundred and eighty-five mRNAs were specifically expressed in the control group, while 198 mRNAs were specifically expressed in the US group (Fig. 2B). Enrichment analysis revealed the ten most activated signaling pathways (Fig. 2C). Among these pathways, the MAPK signaling pathway was the most activated. Furthermore, the expression levels of three factors (Nfkb1, Hras, and Rras2) were significantly upregulated in this pathway.
Fig. 2.
RNA sequencing analysis of myotubes after ultrasound (US) irradiation. A: Volcano plot of differentially expressed RNAs in control group vs. US group. Red dots represent RNAs with statistically significant difference, and blue dots indicate RNAs with no statistically significant difference between the US and control groups. B: A Venn diagram showing unique and overlapping differentially expressed genes (DEGs) between US and control groups. C: Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was performed on differentially expressed RNAs, with the 10 most enriched pathways linked to signaling transduction presented (n = 3). 3.3. US irradiation induces phosphorylation of p38 MAPK in myotubes
To assess MAPK activation at the protein level, we specifically measured the activation of phospho-p38, a key kinase in the MAPK pathway, 24 h after US irradiation in myotubes. Western blotting analysis revealed a significant increase in the phosphorylation level of p38 MAPK (phospho-p38), while the protein levels of GAPDH and total-p38 remained unchanged (Fig. 3A and B, Supplementary File, Fig. 4).
Fig. 4.
Metabolite analysis of myotubes after ultrasound (US) irradiation. A: Overview of enrichment analysis of myotubes after US irradiation. B: Overview of pathway analysis of myotubes after US irradiation. C: The abundance of metabolites in purine metabolism. All values are represented as mean ± SEM.∗p < .05 vs. control (n = 3). 3.4. US irradiation activates purine metabolism in myotubes
To study the metabolite profile of the myotubes after US irradiation, the abundance of metabolites in the myotubes was quantified. To investigate the potential metabolic pathways affected in myotubes after US irradiation, enrichment and pathway analyses were conducted using MetaboAnalyst 5.0 online software (Fig. 4). As depicted in Fig. 4A and B, US irradiation significantly activated several metabolic pathways in myotubes, including pyruvate metabolism, glutamate metabolism, the citric acid cycle, glycogenesis, and purine metabolism, with purine metabolism showing the most significantly activation. Within purine metabolism, the levels of guanosine 5-diphosphate, 3-diphosphate, GDP, deoxyinosine, and adenylosuccinate were significantly elevated, while the expression level of uric acid remained unchanged (Fig. 4C).
3.5. US irradiation enhances mitochondrial respiration in myotubes
To determine the effects of US irradiation on mitochondrial respiration in myotubes, we performed a seahorse assay. Fig. 5 illustrates the changes in the OCR of myotubes upon successive addition of oligomycin, FCCP, rotenone/succinate, and antimycin A during the experiment. Compared to the control group, US irradiation significantly increased the mitochondrial basal respiration rate. In addition, a tendency towards increased maximal mitochondrial respiration was observed.
Fig. 5.
Seahorse analysis of myotubes after ultrasound (US) irradiation. All values are presented as mean ± SEM.∗p < .05 vs. control (n = 3). 4. Discussion
In the present study, we investigated the effects of US irradiation on myotubes. The results of cell viability analysis indicated that US irradiation did not induce cell damage. Furthermore, DNA damage was detected in the myotubes after US irradiation. However, the results indicated that US irradiation did not increase DNA damage, confirming the safety of US, which is consistent with previous research [21].
In the present study, US irradiation activated various types of RNAs, with the MAPK signaling pathway showing the greatest activation. In addition, western blotting revealed that MAPK was activated at the protein level 24 h post-US. US irradiation reportedly affects the MAPK signaling pathway [22]. The findings of this study also indicated that the effects and changes induced by US irradiation in myotubes are mainly mediated through the activation of this pathway. Furthermore, the expression of Nfkb1, Hras, and Rras2 in the MAPK pathway was significantly upregulated. Hras and Rras2 are members of the RAS small GTPase family [23]. RAS interacts with various downstream factors such as PI3K to affect purine metabolism [24]. Activation of the MAPK signaling pathway can upregulate purine biosynthesis by increasing the expression of purine metabolic enzymes [25,26]. Thus, we believe that US stimulates purine metabolism in myotubes through MAPK pathway activation. Purine metabolism encompasses pathways for both the synthesis and degradation of purines, which are key components of cellular energy systems, signaling molecules, and nucleic acids. Disruptions in purine metabolism can affect purine catabolism, nucleotide synthesis, and salvage pathways [27]. Activation of purine metabolism helps meet increased energy demands, ensures a sufficient supply for cells, and enhances intracellular signaling processes, all of which are essential for cell growth, proliferation, and overall metabolic efficiency [27,28]. Uric acid, the end product of purine catabolism, can accumulate and contribute to disorders like gout [27]. In this study, the expression levels of guanosine 5-diphosphate, 3-diphosphate, GDP, deoxyinosine, and adenylosuccinate were significantly increased in myotubes irradiated with US, which is beneficial for cell growth and cellular energy metabolism [29,30]. Simultaneously, uric acid expression did not increase, indicating that US irradiation had no pathological effect on the myotubes despite the activation of purine metabolism.
In contrast, activation of the Ras cascade enhances the mitochondrial content of respiratory enzymes, thereby enhancing respiratory competence [31]. NFkB activation can promote mitochondrial respiration [32]. Our study demonstrated that US irradiation significantly upregulated the expression levels of Nfkb1, Hras, and Rras2 and increased the mitochondrial basal respiration rate in myotubes without increasing DNA damage and ROS expression. This suggests that US irradiation can safely enhance mitochondrial respiration by activating the MAPK signaling pathway in myotubes. Skeletal muscles with higher mitochondrial respiratory capacity can meet increased energy demands during exercise, allowing muscles to sustain higher workloads for longer periods before fatigue sets in Ref. [33]. Furthermore, strong mitochondrial function helps cells manage stresses such as oxidative damage, calcium overload, and metabolic disturbances, offering protection against mitochondrial dysfunction linked to diseases and aging [34,35]. Therefore, we propose that US irradiation has the potential to enhance muscle function.
In addition to the MAPK signaling pathway, the analysis suggests that the Hippo and Wnt signaling pathways were also be activated, as indicated by the changes in mRNA expression levels observed in this study. YAP and Beta Catenin are the predominant factors in these pathways [36], and the expression of YAP was significantly increased in the Hippo signaling pathway in this study (Supplementary File, Fig. 2). While mRNA expression changes can provide insights into the potential activation of these pathways, it is important to note that mRNA levels alone do not directly confirm the activation of the associated protein factors. Previous studies have shown that while YAP and Beta Catenin are transcriptional factors involved in cell growth and can promote neoplastic transformation if deregulated, previous studies have indicated that YAP’s activity may lead to increased nuclear localization and promote cell growth without necessarily causing malignant transformation, unless combined with additional oncogenic signals [36,37]. In summary, we believe that the conditions provided by the US irradiation used in this study were insufficient to induce neoplastic transformation and further experiments, including protein-level analysis, will be necessary to more definitively assess the activation of these pathways.
While the applications of US have traditionally centered on imaging and physiotherapeutic purposes, in this study, our exploration of its influence on muscle cells revealed a previously unknown dimension of cellular responses. We found that US irradiation promoted purine metabolism and mitochondrial respiration by activating the MAPK signaling pathway, benefiting muscle function by supporting cellular energy and protecting against stress. These findings regarding the effects of US on cellular functions significantly contribute to the expanding body of knowledge in this area, offering valuable insights with far-reaching implications for both clinical practice and research applications.
5. Conclusion
This study illustrated the effects of US on cultured myotubes in terms of genomic analysis, metabolomic analysis, and mitochondrial function. US irradiation can trigger the MAPK signaling pathway, thereby activating purine metabolism and improving mitochondrial respiration. Thus, US irradiation is an effective method for boosting skeletal muscle function.
CRediT authorship contribution statement
Xiaoqi Ma: Writing – review & editing, Writing – original draft, Visualization, Methodology, Funding acquisition, Formal analysis, Data curation, Conceptualization. Noriaki Maeshige: Writing – review & editing, Supervision, Resources, Funding acquisition, Formal analysis, Data curation, Conceptualization. Atomu Yamaguchi: Writing – review & editing, Visualization, Methodology, Formal analysis, Data curation, Conceptualization. Yunfei Fu: Writing – review & editing, Visualization, Data curation. Jihao Xing: Writing – review & editing, Visualization, Data curation. Qingcheng Guo: Writing – review & editing, Data curation, Investigation. Hao Lin: Writing – review & editing, Visualization, Data curation. Fuwen Lu: Writing – review & editing, Data curation. Hiroyo Kondo: Writing – review & editing, Resources. Hidemi Fujino: Writing – review & editing, Supervision, Resources, Formal analysis, Data curation.
Mitochondrial dysfunction in muscle cells induced by snoring vibrations
https://www.sciencedirect.com/science/article/pii/S1567724926000644
Highlights
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Snoring vibration disrupts mitochondrial homeostasis in muscle cells.
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Mechanosensing–mechanotransduction axis is activated (integrins, PIEZO1)
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RNA processing defects cause transcript–protein uncoupling.
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Mitochondrial respiration collapses with impaired glycolytic reserve.
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Pusion-dominant remodeling and transient senescence reflect stress adaptation.
Abstract
SVENSKA
2026-08-11 (använder AI för att jag har dyslexi och är ofta för komplicerad i min text/tal MEN jag har granskat detta – samtidigt är det en del som jag gör annorlunda – men liksom alla verktyg så anpassar man användning till var och en MEN utifrån bästa möjliga kunskap eller …
Fördel med att integrera rörelse (yoga eller Chi Kung inre övningar), massage med ljudresonans vid långsam utandning
AI-översikt
Att integrera långsam utandning med rörelse, inre övningar (som yoga eller Chi Kung) och ljudresonans skapar en kraftfull somatisk synergi. Denna kombination utlöser snabbt vagusnerven, förbättrar intern cellulär mikromassage genom ljudvibrationer, förflyttar nervsystemet till djup återhämtning och optimerar kardiovaskulär koherens.
Ohm hälsa +2
Reglering av nervsystemet
- Parasympatisk aktivering: Utdragna, långsamma utandningar signalerar till hjärnan att stänga av ”kamp eller flykt”-responsen och aktivera ”vila och matsmältning”-läge.
BBC +1
- Vagal stimulering: Diafragmans mekaniska rörelse kombinerat med vokaliserad ljudresonans masserar fysiskt vagusnervens banor i bröstkorgen och halsen. BvS-> se också https://www.boaim2.se/clinical-focuses/the-inflammatory-reflex-and-how-transcutaneous-auricular-vagus-nerve-stimulation-may-activate-the-pathway/ där man kan anv’nda t.ex. ”Slå på himmelens trumma” (eller alla inopererade – onödigt? – inplantat på påverkar Vagus via redukulära grenen av Vagus.
SoundSelf +1
- Hjärtfrekvensvariabilitet (HRV): Långsam, rytmisk andning synkroniserar hjärtats och andningsrytmer och förbättrar kroppens interna återkopplingsslingor.
Ohm Health
Fysisk och energimässig integration
- Intern mikromassage: Ljudresonans (såsom nynnande eller toning vid utandning) skickar mjuka tryckvågor genom vävnaden och minskar strukturell spänning.
SoundSelf
- Flytande rörelse: Att kombinera yoga- eller Chi Kung-rörelser med en förlängd utandning hjälper till att frigöra envisa muskelgrepp och ledstelhet.
jingselfcare.com +1
- Förbättrad cirkulation: Taktade rörelser i kombination med reglerad andning stödjer optimal blodflöde och effektivt gasutbyte i lungorna.
Nationella hälsoinstituten (NIH) | (.gov) +1
Mental och emotionell klarhet
- Sinne-kropp-ankare: Det dubbla fokuset på fysisk form och ljud ger ett överaktivt sinne en stabil vilopunkt, vilket fördjupar meditativa tillstånd.
- Stressavlägsnande: Den rytmiska loopen minskar grundläggande psykologisk spänning och minskar produktionen av stresshormoner.
BvS-> Kan avläsas i Respiratorisk Sinus aryrmi (RSA) i labbet
Nationella hälsoinstituten (NIH) | (.gov) +3
AI-fortsättningsförslag:
- Jag ska anpassa mitt nästa svar. Säg bara:
- Chi Kung eller Yogas primära fokus
- Specifik ljudtyp (surrande, tonande eller tystnad)
- Målmål (sömn, stresslindring eller energi)
Ytterligare länkar
Livets andetag: Den respiratoriska vagala stimuleringsmodellen för kontemplativ aktivitet
https://pmc.ncbi.nlm.nih.gov/articles/PMC6189422/
Kan andning med yoga, qigong och tai chi stödja den psykoimmuna homeostasen under och efter covid-19-pandemin? En narrativ granskning
https://pmc.ncbi.nlm.nih.gov/articles/PMC9601556/
Vetenskapen om andning och ljud: Hur resonans reglerar nervsystemet
https://soundself.com/blogs/news/the-science-of-breath-and-sound-how-resonance-regulates-the-nervous-system
Långsam andning för att minska stress: Effekten av att förlänga utandningen ☆
https://pmc.ncbi.nlm.nih.gov/articles/PMC10395759/
Aristoteles: ”tro ej på påståenden innan du kritiskt granskat (BvS-> utifrån ditt eget väldefinierade paradigm) Se också https://www.boaim2.se/innovativm2/some-thought-on-evolution-of-home-sapiens-information-processing/is-purely-rational-human-part-of-our-brain-thinking-the-platform-for-our-contemporary-homo-sapiens-knowledge/thales-and-how-to-increase-understanding-and-more/
BvS-> Huvudfråga just nu
Hur kan jag skapa ljudresonans som underlättar skapandet av optimal enzymatisk påverkan på mitokondrier?
Att skapa direkt ljudresonans för att specifikt optimera individuella mitokondrieenzymer i levande mänsklig vävnad är fortfarande ett experimentellt teoretiskt koncept snarare än en etablerad klinisk praxis. Medan framväxande in vitro-studier tyder på att specifika akustiska vågor och musikaliska frekvenser kan påverka cellulär bioenergetik, mekanotransduktion och ATP-produktion, kräver det precisa fysiska parametrar att rikta in sig på distinkta interna enzymatiska reaktioner via externt ljud.
Biofysiska mekanismer för ljud och celler
Mechanosensing: Akustiska tryckvågor färdas genom mjukvävnader och aktiverar cellytans mekanosensitiva jonkanaler och integriner.
Cytoskeletal transmission: Vibrationer överförs direkt genom cytoskelettet till organellerna, vilket förändrar mitokondriell transport och membranoscillationer.
Bioenergetiska skiften: Kontrollerad lågfrekvent akustisk eller ultraljudsstimulering har observerats i preliminära modeller för att modulera respiratorisk kedjeaktivitet och förbättra ATP-syntesen.
Experimentella metoder utforskade i forskning
Lågfrekventa akustiska fält: Studier som undersöker specifika akustiska vibrationer eller rytmiska tonmönster noterar förändringar i transmembranpotentialstabilitet och cellrespiration.
Pulserat ultraljud: Icke-termiska ultraljudsparametrar med låg intensitet (t.ex. specifika MHz- eller kHz-intervall levererade vid låga arbetscykler) har undersökts för sin förmåga att säkert stimulera metabola vägar utan att orsaka termisk skada.
Vibrationskoherens: Teoretiska modeller antyder att matchning av de naturliga oscillerande frekvenserna hos cellstrukturer kan främja synkronisering av membranpotentialer, även om reproducerbara parametrar för specifika mitokondrieenzymer (som ATP-syntas) ännu inte är standardiserade för mänsklig tillämpning.
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Utforska mitokondriell oscillation och resonans: Inverkan på ATP-produktion och distribution inom hälsa och sjukdomar
https://wms-site.com/alert-on-mitochondria/1154-mitochondrial-oscillations-concept-and-future-perspectives
Vad menar vi med mitokondrieoscillationer och resonans?
Mitokondriella oscillationer och resonans kan verkligen ha betydande konsekvenser för ATP-produktion och distribution. Som cellens primära ”kraftverk” är mitokondrier ansvariga för huvuddelen av ATP-produktionen genom processen oxidativ fosforylering. Effektiviteten i denna process är intimt kopplad till mitokondriernas membranpotential, som genomgår periodiska oscillationer.
Dessa svängningar kan påverka ATP-produktionshastigheten på ett par sätt. När membranpotentialen är hög är mitokondrierna i ett energibesparande tillstånd, och ATP-produktionen är vanligtvis effektiv. Omvänt, när membranpotentialen är låg, är mitokondrierna i ett energiförbrukande tillstånd, och ATP-produktionen kan vara mindre effektiv.
När det gäller ATP-distribution är mitokondrier kända för att bilda dynamiska nätverk inom celler, vilket möjliggör effektiv överföring av ATP och andra metaboliter dit de behövs som mest. Mitokondriernas oscillerande beteende skulle potentiellt kunna spela en roll i regleringen av denna distributionsprocess. Till exempel kan förändringar i det oscillerande tillståndet hos en viss mitokondrie utlösa omfördelningen av ATP inom det mitokondriella nätverket, vilket gör att cellen snabbt kan reagera på förändrade energibehov.
Dessutom skulle konceptet mitokondriell resonans kunna påverka ATP-distributionen ytterligare. Om olika mitokondrier i en cell eller vävnad kan ”resonera” med varandra – det vill säga synkronisera sina svängningar – kan detta möjliggöra en mer samordnad och effektiv distribution av ATP i hela cellen eller vävnaden.
Konceptet mitokondriell resonans är fascinerande och kan ha långtgående konsekvenser för hur energi distribueras i celler och vävnader.
Som en analogi kan vi betrakta hur synkronsimmare kan röra sig mer effektivt genom vatten genom att koordinera sina rörelser. På liknande sätt, om olika mitokondrier i en cell eller vävnad kan ”resonera” med varandra – det vill säga synkronisera sina svängningar – kan detta möjliggöra en mer koordinerad och effektiv distribution av ATP i hela cellen eller vävnaden.
När mitokondrier är i resonans synkroniseras svängningarna i deras membranpotentialer. Denna synkronisering kan underlätta bildandet av ett mer effektivt energidistributionsnätverk, vilket gör att ATP och andra metaboliter snabbt och effektivt kan transporteras dit de behövs mest i cellen eller vävnaden.
Detta kan vara särskilt viktigt i celler med höga energibehov, såsom nervceller eller muskelceller, eller i situationer där energibehovet kan förändras snabbt, såsom under träning eller som svar på stress. Resonans kan göra det möjligt för dessa celler att snabbt öka ATP-produktionen och distributionen som svar på ökat energibehov, vilket säkerställer att alla delar av cellen har tillgång till den energi de behöver för att fungera optimalt.
Mitokondriell oscillation – Projektets mål
Vårt mål är att fördjupa sig i de komplexa biologiska fenomenen mitokondriell dynamik – oscillationer (kontinuerliga förändringar i mitokondriemembranpotential) och resonans (förstärkning av effekterna av en extern kraft som matchar ett systems egen naturliga frekvens) – och deras inflytande på det cellulära energilandskapet.
Dessa oscillationer spelar en central roll i cellulär signalering, energimetabolism och produktion av reaktiva syreradikaler, med förändringar kopplade till olika patologiska tillstånd såsom neurodegenerativa sjukdomar och cancer. Dessutom förbättrar mitokondriell resonans potentiellt intermitokondriell och cellulär kommunikation, vilket optimerar energiöverföring och signalvägar.
En viktig aspekt av detta projekt fokuserar på konsekvenserna av dessa processer för ATP-produktion och distribution. Mitokondrier, som cellulära kraftverk, är primärt ansvariga för ATP-produktion via oxidativ fosforylering.
Detta projekt kommer att utforska:
Hur förändringar i mitokondriemembranpotential under oscillationer påverkar effektiviteten i ATP-produktionen .
- Hur mitokondriell nätverksdynamik, reglerad av oscillerande beteende, kan påverka ATP-distributionen i cellen.
- Hur mitokondriell resonans kan möjliggöra en mer samordnad och effektiv distribution av ATP i celler eller vävnader. Konceptet mitokondriell resonans kan ytterligare påverka ATP-distributionen. Om olika mitokondrier i en cell eller vävnad kan ”resonera” med varandra – det vill säga synkronisera sina oscillationer – kan detta möjliggöra en mer samordnad och effektiv distribution av ATP i hela cellen eller vävnaden.