Intracellular Stimulus Research Articles

Arresting the G2/M phase empowers synergy in magnetic nanomanipulator-based cancer mechanotherapy and chemotherapy

Using mechanical cues for cancer cells can realize precise control and efficient therapeutic effects. However, the cell cycle-specific response for dynamic mechanical manipulation is barely investigated. Here, RGD-modified iron oxide nanomanipulators were utilized as the intracellular magneto-mechanical transducers to investigate the mechanical impacts on the cell cycle under a dynamic magnetic field for cancer treatment. The G2/M phase was identified to be sensitive to the intracellular magneto-mechanical modulation with a synergistic treatment effect between the pretreatment of cell cycle-specific drugs and the magneto-mechanical destruction, and thus could be an important mechanical-targeted phase for regulation of cancer cell death. Finally, combining the cell cycle-specific drugs with magneto-mechanical manipulation could significantly inhibit glioma and breast cancer growth in vivo. This intracellular mechanical stimulus showed cell cycle-dependent cytotoxicity and could be developed as a spatiotemporal therapeutic modality in combination with chemotherapy drugs for treating deep-seated tumors.

Functional immunophenotyping of children with critical status asthmaticus identifies differential gene expression responses in neutrophils exposed to a poly(I:C) stimulus

The host immune response to a viral immune stimulus has not been examined in children during a life-threatening asthma attack. We determined whether we could identify clusters of children with critical asthma by functional immunophenotyping using an intracellular viral analog stimulus. We performed a single-center, prospective, observational cohort study of 43 children ages 6–17 years admitted to a pediatric intensive care unit for an asthma attack between July 2019 to February 2021. Neutrophils were isolated from children, stimulated overnight with LyoVec poly(I:C), and mRNA was analyzed using a targeted Nanostring immunology array. Network analysis of the differentially expressed transcripts for the paired LyoVec poly(I:C) samples was performed. We identified two clusters by functional immunophenotyping that differed by the Asthma Control Test score. Cluster 1 (n = 23) had a higher proportion of children with uncontrolled asthma in the four weeks prior to PICU admission compared with cluster 2 (n = 20). Pathways up-regulated in cluster 1 versus cluster 2 included chemokine receptor/chemokines, interleukin-10 (IL-10), IL-4, and IL-13 signaling. Larger validation studies and clinical phenotyping of children with critical asthma are needed to determine the predictive utility of these clusters in a larger clinical setting.

Macromolecular Crowding as an Intracellular Stimulus for Responsive Nanomaterials.

Stimuli-responsive materials are exploited in biological, materials, and sensing applications. We introduce a new endogenous stimulus, biomacromolecule crowding, which we achieve by leveraging changes in thermoresponsive properties of polymers upon high concentrations of crowding agents. We prepare poly(2-oxazoline) amphiphiles that exhibit lower critical solution temperatures (LCST) in serum above physiological temperature. These amphiphiles stabilize oil-in-water nanoemulsions at temperatures below the LCST but are ineffective surfactants above the LCST, resulting in emulsion fusion. We find that the transformations observed upon heating nanoemulsions above their surfactant's LCST can instead be induced at physiological temperatures through the addition of polymers and protein, rendering thermoresponsive materials "crowding responsive." We demonstrate that the cytosol is a stimulus for nanoemulsions, with droplet fusion occurring upon injection into cells of living zebrafish embryos. This report sets the stage for classes of thermoresponsive materials to respond to macromolecule concentration rather than temperature changes.

Autophagy modulation by viruses: An important role in tumor progression

Autophagy is an important process for cellular homeostasis at critical steps of development or in response to environmental stress. In the context of cancers, autophagy has a significant impact on tumor occurrence and tumor cell growth. On the one hand, autophagy limits the transformation of precancerous cells into cancer cells at an early stage. However, on the other hand, it promotes cell survival, cell proliferation, metastasis and resistance to anti-tumor therapies in more advanced tumors. Autophagy can be induced by a variety of extracellular and intracellular stimulus. Viral infections have often been associated with a modulation of autophagy, with variable impacts on viral replication and on the survival of infected cells depending on the model studied. In a tumor context, the modulation of autophagy induced by the viral infection of tumor cells seems to have a significant impact on tumor progression. The aim of this review article is to present recent findings regarding the consequences of autophagy disturbance by viral infections on tumor behavior.

Intracellular imaging and concurrent pH sensing of cancer-derived exosomes using surface-enhanced Raman scattering.

Exosomes have attracted significant attention as cancer diagnostic targets and therapeutic agents due to their unique biogenesis and structure. To clarify the biological activities of exosomes, it is important to obtain a picture of their intracellular distribution and how they evolve over time. In this work, a new kind of intracellular exosome imaging and concurrent pH sensing method is demonstrated by using the surface-enhanced Raman scattering (SERS) technique. Specifically, 4-mercaptobenzoic acid (4MBA)-tagged silver nanoparticles are attached onto the outer surfaces of exosomes, in which silver nanoparticles are employed as SERS generators. Raman agents 4MBA are susceptible to a specific intracellular stimulus, that is, undergo a protonation or deprotonation in response to intracellular pH variation, which correspondingly exhibit different vibrational spectra features. By using the SERS spectroscopy, tracking of the intracellular distribution of exosomes and the concurrent quantitative sensing of environmental pH were achieved, which demonstrated that, as time prolonged, exosomes first attached with the tumor cell surfaces, and then entered into the cells and accumulated in lysosomes. Such SERS-active hybridized exosomes, that are sensitive to discrete variations in intracellular pH, have proved their capability for the investigation of interactions between exosomes and cells. The spectral diversity and flexible surface modification of these hybridized exosomes are also highly expected in developing multifunctional exosome-based nanoplatforms, which offers great potential to promote the exosome-based therapeutics forward into an advanced stage.

Bioreducible Zinc(II)-Dipicolylamine Functionalized Hyaluronic Acid Mediates Safe siRNA Delivery and Effective Glioblastoma RNAi Therapy.

RNA interference (RNAi) is an emerging therapeutic modality for tumors. However, lack of a safe and efficient small interfering RNA (siRNA) delivery system limits its clinical application. Here, we report a bioreducible and less-cationic siRNA delivery carrier by conjugating Zn(II)-dipicolylamine complexes (Zn-DPA) onto hyaluronic acid (HA) via a redox-sensitive disulfide (-SS-) linker. Such polymer conjugates can formulate stable siRNA nanomedicines via coordination between zinc ions of DPA and the anionic phosphate of siRNA. After the conjugates are taken up by cells, intracellular reduction stimulus subsequently triggers the release of siRNAs and elucidates the desired RNAi effect. Our studies showed the formulated siRNA nanomedicines can be efficiently delivered into tumor cells/tissues and mediates less cytotoxicities both in vitro and in vivo. More importantly, when applied in a xenograft glioblastoma tumor model, this siRNA nanomedicine demonstrated significantly enhanced antitumor ability comparing to naked siRNA. This work demonstrates that such bioreducible Zn-DPA-functionalized HA conjugates without using cationic material as a siRNA carrier represents a promising direction for RNAi-based cancer therapy.

Comparison of intracellular location and stimulus reaction times of forisomes in sieve tubes of four legume species

ABSTRACTForisomes in legumes are responsible for fast sieve-element occlusion in response to injury to the vascular system. This prevents uncontrolled leakage of phloem sap and protects against invasion of pathogens. Here we compared forisomes of four different legumes (Pisum sativum, Vicia faba, Trifolium pratense and Medicago sativa) by their location (basal, central, apical) in the sieve element and reactivity to a distant heat stimulus. In each species, the majority of forisomes was located basally. Yet, we found differences in intracellular location: forisomes are distributed more evenly in the sieve elements of Pisum. After burning, basally located forisomes of the four species reacted with dispersion, followed by a spontaneous recondensation with similar reaction times. The results suggest universal forisome behaviour in fabacean species.

Redox sensitive cationic pullulan for efficient gene transfection and drug retention in C6 glioma cells

Thiolated cationic pullulan was synthesized by conjugating pullulan with polyethyleneimine (PEI) and mercaptosuccinic acid (MSA). The formed conjugate was oxidized to obtain disulfide linked cationic pullulan (PPMSS). PPMSS exhibited good buffering capacity and nanoplexes formulated were of size less than 150nm. Nanoplexes formed with PPMSS are redox sensitive and susceptible to reductive cleavage by dithiothreitol (DTT) ensuring the intracellular release of DNA. In vitro, cytotoxic evaluation studies of polymers in C6 cell lines established its non-toxic nature. The studies using endocytosis inhibitors revealed the uptake pathways of nanoplexes. Further, the plasmid and polymer tracking studies indicated the successful unpacking of DNA from the nanoplexes and its nuclear localization. The gene transfection efficiency was established by the p53 gene expression studies. Furthermore, the ability of the polymer to inhibit efflux pumping in cancer cells has also been elucidated in terms of P-gp inhibition studies and drug retention kinetics using the anticancer drug, doxorubicin (DOX). Our results also suggest that greater retention of DOX was accompanied by the reduction of disulfide linkage by a ubiquitous intracellular stimulus, glutathione. Thus simultaneous gene and drug delivery using redox sensitive cationic polymers may have a promising potential in cancer therapy.

General Approach of Stimuli-Induced Aggregation for Monitoring Tumor Therapy.

Intracellular construction of nanoaggregates from synthetic molecules to mimic natural ordered superstructures has gained increasing attention recently. Here, we develop an endogenous stimuli-induced aggregation (eSIA) approach to construct functional nanoaggregates for sensing and monitoring cellular physiological processes in situ. We design a series of thermosensitive polymer-peptide conjugates (PPCs), which are capable of constructing nanoaggregates in cells based on their isothermal phase transition property. The PPCs are composed of three moieties (i.e., a thermoresponsive polymer backbone, a grafted peptide, and a signal-molecule label). The bioenvironment-associated phase transition behavior of PPCs are carefully studied by consideration of various crucial parameters such as chain length, hydrophilicity, ratio of grafted peptides, and concentration. Intriguingly, under the specific intracellular stimulus, the PPCs are tailored and simultaneously form nanoaggregates exhibiting long-term retention effect, which enables specific identification and quantification of endogenous factors. This general approach is expected for high-performance in situ sensing and dynamic monitoring of disease progression in living subjects.

Metabolic perturbation of epigenome by inhibiting S-adenosylhomocysteine hydrolase elicits senescence through DNA damage response in hepatoma cells.

Cellular senescence is a key physiological barrier against tumor and represents an option for therapeutic intervention. One pivotal intracellular stimulus causing senescence is DNA damage response, while the senescence-associated heterochromatin in cancer limits the strength of the DNA damage response to endogenous genotoxic stress or DNA-damaging agents. Therefore, targeting the maintenance of compacted chromatin in cancer cells represents an optional intervention to improve the therapeutic efficacy in cancer treatment. Given a crosstalk between methionine cycle and histone methylation, we hypothesize that pharmacologically disrupting methylation potential, defined as the ratio of cellular S-adenosylmethionine to S-adenosylhomocysteine, could affect the chromatin structures in cancer cells and thus enhance their sensitivity to DNA damage response signaling. Our results showed that 3-deazaneplanocin A, a chemical inhibitor of S-adenosylhomocysteine hydrolase, elicited a typical cellular senescence in hepatoma cells. Therapy-induced senescence by 3-deazaneplanocin A was mediated through p53-p21 pathway and triggered by enhanced ataxia-telangiectasia mutated activation related to chromatin changes. In conclusion, our study demonstrated that metabolic perturbation of chromatin status in oncogene-activated cancers could be an optional intervention to sensitize DNA damage response signaling.

Dual responsive nanogels for intracellular doxorubicin delivery

Nanosized polymeric delivery systems that encapsulate drug molecules and release them in response to a specific intracellular stimulus are of promising interest for cancer therapy. Here, we demonstrated a simple and fast synthetic protocol of redox-responsive nanogels with high drug encapsulation efficiency and stability. The prepared nanogels displayed narrow size distributions and versatility of surface modification. The polymer precursor of these nanogels is based on a random copolymer that contains oligoethyleneglycol (OEG) and pyridyldisulfide (PDS) units as side-chain functionalities. The nanogels were prepared through a lock-in strategy in aqueous media via self cross-linking of PDS groups. By changing polymer concentration, we could control the size of nanogels in range of 80⿿115nm. The formed nanogels presented high doxorubicin (DOX) encapsulation efficiency (70% (w/w)) and displayed pH and redox-controlled drug release triggered by conditions mimicking the reducible intracellular environment. The nanogels displayed an excellent cytocompatibility and were effectively endocytosed by A2780CP ovarian cancer cells, which make them promising nanomaterials for the efficient intracellular delivery of anticancer drugs.

An in vitro investigation of a detachable fork-like structure as efficient nuclear-targeted sub-unit in A2780 cell cultures

The pharmacological target of many anticancer drugs is those molecules associated with genetic information which are localized in nucleus. To efficiently deliver drugs into cancer cell nucleus, in our previous study, a fork-like sub-unit, with one end conjugated with a targeting peptide named R8NLS (CRRRRRRRRPKKKRKV) and the other end conjugated with c-Myc oncogene inhibitor H1-S6A,F8A (H1) peptide, was linked onto the N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer via an enzyme degradable glycylphenylalanylleucylglycine (GFLG) tetrapeptide spacer. Here, an in vitro mechanism investigation of the fork-like sub-unit was studied in detail. We found that the fusion with two complementary R8 and NLS motifs is required to exert the multifunctional targeting capability of the tandem R8NLS peptide in overcoming various intracellular barriers, including enhancing cellular uptake, facilitating endosomal escape and penetrating through the double-layered nuclear membrane. Also required is the tactful detachment of the fork-like sub-unit from the copolymer in response to intracellular stimulus, because a smaller sub-unit not only increases the intracellular trafficking efficiency by reducing the size burden of magical bullet R8NLS, but also guarantees successful entry through the restricted nucleopore. Herein, this study highlights that both nuclear targeting ligand R8NLS and detachable fork-like sub-unit are dispensable for programmed nuclear delivery and may show feasibility on other drug delivery systems, such as nanoparticles and micelles.

Similar Intracellular Location and Stimulus Reactivity, but Differential Mobility of Tailless (Vicia faba) and Tailed Forisomes (Phaseolus vulgaris) in Intact Sieve Tubes

Sieve elements of legumes contain forisomes—fusiform protein bodies that are responsible for sieve-tube occlusion in response to damage or wound signals. Earlier work described the existence of tailless and tailed forisomes. This study intended to quantify and compare location and position of tailless (in Vicia faba) and tailed (in Phaseolus vulgaris) forisomes inside sieve elements and to assess their reactivity and potential mobility in response to a remote stimulus. Location (distribution within sieve elements) and position (forisome tip contacts) of more than altogether 2000 forisomes were screened in 500 intact plants by laser scanning confocal microscopy in the transmission mode. Furthermore, we studied the dispersion of forisomes at different locations in different positions and their positional behaviour in response to distant heat shocks. Forisome distribution turned out to be species-specific, whereas forisome positions at various locations were largely similar in bushbean (Phaseolus) and broadbean (Vicia). In general, the tailless forisomes had higher dispersion rates in response to heat shocks than the tailed forisomes and forisomes at the downstream (basal) end dispersed more frequently than those at the upstream end (apical). In contrast to the tailless forisomes that only oscillate in response to heat shocks, downstream-located tailed forisomes can cover considerable distances within sieve elements. This displacement was prevented by gentle rubbing of the leaf (priming) before the heat shock. Movement of these forisomes was also prohibited by Latrunculin A, an inhibitor of actin polymerization. The apparently active mobility of tailed forisomes gives credence to the idea that at least the latter forisomes are not free-floating, but connected to other sieve-element structures.

Knockdown of interleukin-1α does not attenuate LPS-induced production of interleukin-1β in mouse macrophages

IL-1α and IL-1β are synthesized as 31kDa cell-associated precursors following TLR-4 stimulation, but their processing to the mature form and secretion require a second intracellular stimulus. The unique localization of the precursor of IL-1α (pro-IL-1α) to the nucleus suggested a role in transcriptional regulation of inflammatory cytokines. We explored the hypothesis that pro-IL-1α is involved in regulation of IL-1β expression following TLR-4 stimulation. IL-1β mRNA and protein levels were specifically decreased in macrophages from IL-1α-deficient mice following TLR-1/2, TLR-4 or TLR-9 stimulation, supporting the hypothesis. However, activation of the main upstream regulators of IL-1β expression, IRF3, NFkB and p38/JNK, were not reduced in macrophages from IL-1α-deficient mice. In order to assess the specific role of IL-1α in macrophages, we generated mice with myeloid cell deficiency of IL-1α (LyzMCre-loxp). Despite over 90% knockdown of IL-1α, TLR-4 stimulated macrophages from LyzMCre-loxp mice did not produce lower levels of IL-1β compared to IL-1α-loxp-flanked mice. In order to overcome the possibility that effects are caused by the incomplete deficiency of IL-1α, we generated new whole-body IL-1α knockout mice (GeneralCre-IL-1α) and the findings were similar to myeloid cell-deficient IL-1α. Collectively, our findings do not support the previously suggested role of nuclear IL-1α in gene regulation of IL-1β. Rather, they suggest that IL-1α acts mainly as an alarmin that is sequestered in the nucleus following stimulation with TLR-4.

Remote-controlled release of DNA in living cells via simultaneous light and host-guest mediations.

Using photons as external triggers to realize remote-controlled release of oligonucleotide is superior to other intracellular or external stimulus. UV light is a valid photon-controlled manner due to high efficiency. However, further applications of these approaches in living cells are hampered by the large dose of UV-light irradiation. To address this issue, a simultaneous light and host/guest mediation was proposed in this paper. Gold nanoparticles (AuNPs) encoding with mercapto-β-cyclodextrin (βCD) served as a carried agent. Azobenzene (Azo), which was labeled on a releasing oligonucleotide, acted as a photochemically controlled switch. Ferrocene (Fc), an excellent guest for inclusion complexation by βCD, serves as "enhancers" and shifts the equilibrium of the inclusion-exclusion process between trans-Azo and βCD under UV-light irradiation, thus making the dose of UV-light irradiation reduced obviously. For further application, transfected green fluorescent protein (GFP)-expressing human lung cancer A549 cells were used to determine cellular uptake and gene silencing mediated by our constructed system in vivo. The results demonstrate that by employing Fc host-guest interaction, about 62.4% gene silencing was achieved within 30 min, which is significantly higher than that without Fc competition. Our strategy provides the potential for orthogonal DNA delivery and therapeutic activation that would be capable of achieving higher levels of site-specific activity and reduced amounts of side effects.

Critical role of arachidonic acid-activated mTOR signaling in breast carcinogenesis and angiogenesis

The mammalian target of rapamycin (mTOR) signaling pathway is upregulated in the pathogenesis of many cancers. Arachidonic acid (AA) and its metabolites play critical role in the development of breast cancer, but the mechanisms through which AA promotes mammary tumorigenesis and progression are poorly understood. We found that the levels of AA and cytosolic phospholipase A2 (cPLA2) strongly correlated with the signaling activity of mTORC1 and mTORC2 as well as the expression levels of vascular epithelial growth factor (VEGF) in human breast tumor tissues. In cultured breast cancer cells, AA effectively activated both mTOR complex 1 (mTORC1) and mTORC2. Interestingly, AA-stimulated mTORC1 activation was independent of amino acids, phosphatidylinositol 3-kinase (PI3-K) and tuberous sclerosis complex 2 (TSC2), which suggests a novel mechanism for mTORC1 activation. Further studies revealed that AA stimulated mTORC1 activity through destabilization of mTOR-raptor association in ras homolog enriched in brain (Rheb)-dependent mechanism. Moreover, we showed that AA-stimulated cell proliferation and angiogenesis required mTOR activity and that the effect of AA was mediated by lipoxygenase (LOX) but not cyclooxygenase-2 (COX-2). In animal models, AA-enhanced incidences of rat mammary tumorigenesis, tumor weights and angiogenesis were inhibited by rapamycin. Our findings suggest that AA is an effective intracellular stimulus of mTOR and that AA-activated mTOR plays critical roles in angiogenesis and tumorigenesis of breast cancer.

Postexercise Carbohydrate–Protein Supplementation Improves Subsequent Exercise Performance and Intracellular Signaling for Protein Synthesis

Postexercise carbohydrate-protein (CHO + PRO) supplementation has been proposed to improve recovery and subsequent endurance performance compared to CHO supplementation. This study compared the effects of a CHO + PRO supplement in the form of chocolate milk (CM), isocaloric CHO, and placebo (PLA) on recovery and subsequent exercise performance. Ten cyclists performed 3 trials, cycling 1.5 hours at 70% VO₂max plus 10 minutes of intervals. They ingested supplements immediately postexercise and 2 hours into a 4-hour recovery. Biopsies were performed at recovery minutes 0, 45, and 240 (R0, R45, REnd). Postrecovery, subjects performed a 40-km time trial (TT). The TT time was faster in CM than in CHO and in PLA (79.43 ± 2.11 vs. 85.74 ± 3.44 and 86.92 ± 3.28 minutes, p ≤ 0.05). Muscle glycogen resynthesis was higher in CM and in CHO than in PLA (23.58 and 30.58 vs. 7.05 μmol·g⁻¹ wet weight, p ≤ 0.05). The mammalian target of rapamycin phosphorylation was greater at R45 in CM than in CHO or in PLA (174.4 ± 36.3 vs. 131.3 ± 28.1 and 73.7 ± 7.8% standard, p ≤ 0.05) and at REnd in CM than in PLA (94.5 ± 9.9 vs. 69.1 ± 3.8%, p ≤ 0.05). rpS6 phosphorylation was greater in CM than in PLA at R45 (41.0 ± 8.3 vs. 15.3 ± 2.9%, p ≤ 0.05) and REnd (16.8 ± 2.8 vs. 8.4 ± 1.9%, p ≤ 0.05). FOXO3A phosphorylation was greater at R45 in CM and in CHO than in PLA (84.7 ± 6.7 and 85.4 ± 4.7 vs. 69.2 ± 5.5%, p ≤ 0.05). These results indicate that postexercise CM supplementation can improve subsequent exercise performance and provide a greater intracellular signaling stimulus for PRO synthesis compared to CHO and placebo.

Stimulus Protocol Determines the Most Computationally Efficient Preconditioner for the Bidomain Equations

The efficient solution of the bidomain equations is a fundamental tool in the field of cardiac electrophysiology. When choosing a finite element discretization of the coupled system, one has to deal with the solution of a large, highly sparse system of linear equations. The conjugate gradient algorithm, along with suitable preconditioning, is the natural choice in this scenario. In this study, we identify the optimal preconditioners with respect to both stimulus protocol and mesh geometry. The results are supported by a comprehensive study of the mesh-dependence properties of several preconditioning techniques found in the literature. Our results show that when only intracellular stimulus is considered, incomplete LU factorization remains a valid choice for current cardiac geometries. However, when extracellular shocks are delivered to tissue, preconditioners that take into account the structure of the system minimize execution time and ensure mesh-independent convergence.

Differential induction of apoptosis in human colonic carcinoma cells (Caco-2) by Atopobium, and commensal, probiotic and enteropathogenic bacteria: Mediation by the mitochondrial pathway

The induction of apoptosis in mammalian cells by bacteria is well reported. This process may assist infection by pathogens whereas for non-pathogens apoptosis induction within carcinoma cells protects against colon cancer. Here, apoptosis induction by a major new gut bacterium, Atopobium minutum, was compared with induction by commensal ( Escherichia coli K-12 strains), probiotic ( Lactobacillus rhamnosus, Bifidobacterium latis) and pathogenic ( E. coli: EPEC and VTEC) gut bacteria within the colon cancer cell line, Caco-2. The results show a major apoptotic effect for the pathogens, mild effects for the probiotic strains and A. minutum, but no effect for commensal E. coli. The mild apoptotic effects observed are consistent with the beneficial roles of probotics in protection against colon cancer and suggest, for the first time, that A. minutum possesses similar advantageous, anti-cancerous activity. Although bacterial infection increased Caco-2 membrane FAS levels, caspase-8 was not activated indicating that apoptosis is FAS independent. Instead, in all cases, apoptosis was induced through the mitochondrial pathway as indicated by BAX translocation, cytochrome c release, and caspase-9 and -3 cleavage. This suggests that an intracellular stimulus initiates the observed apoptosis responses.

Extremely Low-Frequency Magnetic Fields Effects on the Snail Single Neurons

The aim of present work is to explore the influence of extremely low-frequency electromagnetic fields (8.34 and 217 Hz) utilized in cell phones on habituation of the mollusk single neuron to intracellular stimuli. The isolated nervous system of the mollusk Helix Pomatia was used in the experiments. Helmholtz coils were used to expose brain ganglia to the low-frequency electromagnetic fields. Peak values of the extremely low-frequency fields were between 1 and 6 mT. Neuron electrophysiology was investigated using a standard microelectrode technique. Exposure of the neuron to the low-frequency electromagnetic fields caused dehabituation to intracellular stimulus. The effect was proportional to the magnetic induction peak value. The observed dehabituation occurs by degradation of the signal to noise ratio and by alteration of the neuron's normal function.